WorldWideScience

Sample records for bohrium 264

  1. Bohrium-A New Element in the Periodic Table

    Indian Academy of Sciences (India)

    Home; Journals; Resonance – Journal of Science Education; Volume 5; Issue 5. Bohrium - A New Element in the Periodic Table. Srinivasan Natarajan. Research News Volume 5 Issue 5 May 2000 pp 95-100. Fulltext. Click here to view fulltext PDF. Permanent link: http://www.ias.ac.in/article/fulltext/reso/005/05/0095-0100 ...

  2. Properties of an α Particle in a Bohrium 270 Nucleus under the Generalized Symmetric Woods-Saxon Potential

    Directory of Open Access Journals (Sweden)

    Bekir Can LÜTFÜOĞLU

    2017-04-01

    Full Text Available The energy eigenvalues and the wave functions of an α particle in a Bohrium 270 nucleus have been calculated by solving Schrödinger equation for Generalized Symmetric Woods-Saxon potential. Using the energy spectrum by excluding and including the quasi-bound eigenvalues, entropy, internal energy, Helmholtz energy, and specific heat, as functions of reduced temperature have been calculated. Stability and emission characteristics have been interpreted in terms of the wave and thermodynamic functions. The kinetic energy of a decayed α particle was calculated using the quasi-bound states, which has been found close to the experimental value.

  3. 40 CFR 264.271 - Treatment program.

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 25 2010-07-01 2010-07-01 false Treatment program. 264.271 Section 264... Treatment § 264.271 Treatment program. (a) An owner or operator subject to this subpart must establish a land treatment program that is designed to ensure that hazardous constituents placed in or on the...

  4. 43 CFR 26.4 - Legislation.

    Science.gov (United States)

    2010-10-01

    ... 43 Public Lands: Interior 1 2010-10-01 2010-10-01 false Legislation. 26.4 Section 26.4 Public Lands: Interior Office of the Secretary of the Interior GRANTS TO STATES FOR ESTABLISHING YOUTH CONSERVATION CORPS PROGRAMS § 26.4 Legislation. State programs must meet all of the requirements of section 4...

  5. 40 CFR 264.14 - Security.

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 25 2010-07-01 2010-07-01 false Security. 264.14 Section 264.14... Standards § 264.14 Security. (a) The owner or operator must prevent the unknowing entry, and minimize the...) for discussion of security requirements at disposal facilities during the post-closure care period...

  6. H.264/AVC Video Compression on Smartphones

    Science.gov (United States)

    Sharabayko, M. P.; Markov, N. G.

    2017-01-01

    In this paper, we studied the usage of H.264/AVC video compression tools by the flagship smartphones. The results show that only a subset of tools is used, meaning that there is still a potential to achieve higher compression efficiency within the H.264/AVC standard, but the most advanced smartphones are already reaching the compression efficiency limit of H.264/AVC.

  7. 40 CFR 264.1065 - Reporting requirements.

    Science.gov (United States)

    2010-07-01

    ... Air Emission Standards for Equipment Leaks § 264.1065 Reporting requirements. (a) A semiannual report...). (iii) The equipment identification number of each compressor for which a leak was not repaired as... monitoring required by § 264.1060 and was not corrected within 24 hours, the duration and cause of each...

  8. 40 CFR 264.341 - Waste analysis.

    Science.gov (United States)

    2010-07-01

    ... the waste feed sufficient to provide all information required by § 270.62(b) or § 270.19 of this... the owner or operator must conduct sufficient waste analysis to verify that waste feed to the... 40 Protection of Environment 25 2010-07-01 2010-07-01 false Waste analysis. 264.341 Section 264...

  9. 40 CFR 264.195 - Inspections.

    Science.gov (United States)

    2010-07-01

    ... wells) to ensure that the tank system is being operated according to its design. [Note: Section 264.15(c... American Petroleum Institute (API) Publication 1632, “Cathodic Protection of Underground Petroleum Storage...

  10. 40 CFR 264.1089 - Recordkeeping requirements.

    Science.gov (United States)

    2010-07-01

    ... tank design features and facility operating procedures currently used to prevent an undue safety hazard... procedures currently used to prevent an undue safety hazard during the management of this hazardous waste in... compounds manufactured at the facility that meet the conditions specified in § 264.1080(d)(1). (2) A...

  11. 40 CFR 264.1053 - Standards: Compressors.

    Science.gov (United States)

    2010-07-01

    .... (b) Each compressor seal system as required in paragraph (a) of this section shall be: (1) Operated... control device that complies with the requirements of § 264.1060, or (3) Equipped with a system that...-vent system capable of capturing and transporting any leakage from the seal to a control device that...

  12. 38 CFR 26.4 - Policy.

    Science.gov (United States)

    2010-07-01

    ... THE DEPARTMENT OF VETERANS AFFAIRS (VA) ACTIONS § 26.4 Policy. (a) VA must act with care in carrying... choice; (3) Achieve a balance between the use and development of resources, within the sustained capacity of the ecological system involved; and, (4) Enhance the quality of renewable resources while working...

  13. 40 CFR 264.1035 - Recordkeeping requirements.

    Science.gov (United States)

    2010-07-01

    ... required by § 264.1033(k). (iii) If engineering calculations are used, a design analysis, specifications... engineering texts -acceptable to the Regional Ad-mini-stra-tor that present basic control device design... regeneration cycle, duration of the carbon bed steaming and cooling/drying cycles, design carbon bed...

  14. 34 CFR 75.264 - Transfers among budget categories.

    Science.gov (United States)

    2010-07-01

    ... 34 Education 1 2010-07-01 2010-07-01 false Transfers among budget categories. 75.264 Section 75.264 Education Office of the Secretary, Department of Education DIRECT GRANT PROGRAMS How Grants Are Made Miscellaneous § 75.264 Transfers among budget categories. A grantee may, notwithstanding any...

  15. 40 CFR 264.232 - Air emission standards.

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 25 2010-07-01 2010-07-01 false Air emission standards. 264.232 Section 264.232 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) SOLID WASTES (CONTINUED) STANDARDS FOR OWNERS AND OPERATORS OF HAZARDOUS WASTE TREATMENT, STORAGE, AND DISPOSAL FACILITIES Surface Impoundments § 264.232 Air emission...

  16. 40 CFR 264.179 - Air emission standards.

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 25 2010-07-01 2010-07-01 false Air emission standards. 264.179 Section 264.179 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) SOLID WASTES (CONTINUED) STANDARDS FOR OWNERS AND OPERATORS OF HAZARDOUS WASTE TREATMENT, STORAGE, AND DISPOSAL FACILITIES Use and Management of Containers § 264.179 Air...

  17. 40 CFR 264.200 - Air emission standards.

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 25 2010-07-01 2010-07-01 false Air emission standards. 264.200 Section 264.200 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) SOLID WASTES (CONTINUED) STANDARDS FOR OWNERS AND OPERATORS OF HAZARDOUS WASTE TREATMENT, STORAGE, AND DISPOSAL FACILITIES Tank Systems § 264.200 Air emission standards....

  18. 36 CFR 264.11 - Use of symbol.

    Science.gov (United States)

    2010-07-01

    ... 36 Parks, Forests, and Public Property 2 2010-07-01 2010-07-01 false Use of symbol. 264.11 Section... MANAGEMENT Mount St. Helens National Volcanic Monument Symbol § 264.11 Use of symbol. Except as provided in § 264.12, use of the Mount St. Helens National Volcanic Monument official symbol, including a facsimile...

  19. 40 CFR 264.276 - Food-chain crops.

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 25 2010-07-01 2010-07-01 false Food-chain crops. 264.276 Section 264... Treatment § 264.276 Food-chain crops. The Regional Administrator may allow the growth of food-chain crops in... Regional Administrator will specify in the facility permit the specific food-chain crops which may be grown...

  20. New isotope 264Sg and decay properties of 262-264Sg

    Energy Technology Data Exchange (ETDEWEB)

    Gregorich, K.E.; Gates, J.M.; Duellmann, Ch.E.; Sudowe, R.; Nelson, S.L.; Garcia, M.A.; Dragojevic, I.; Folden, C.M. III.; Neumann,S.H.; Hoffman, D.C.; Nitsche, H.

    2006-06-20

    New isotope, 264Sg, was identified using the38U(30Si,xn)268-xSg reaction and excitation functions for 262-264Sg weremeasured. 264Sg decays by spontaneous fission with a half life of 37+27/-11 ms. The spontaneous fission branch for 0.9-s 263Sg was measuredfor the first time and found to be (13+-8) percent. 262Sg decays byspontaneous fission with a 15 +5/-3 ms half-life. Spontaneous fissionpartial half-life systematics are evaluated for even-even Sg isotopesfrom 258Sg through 266Sg, spanning the transition region between theN=152, Z=100 and N=162, Z=108 deformed shells.

  1. Bohrium-A New Element in the Periodic Table

    Indian Academy of Sciences (India)

    The periodic table of elements, the basic and most important component of research in chemistry and physics is growing continu- ously. It is interesting to note that until the. 16th century, only a handful of elements have been known to mankind (10, to be pre- cise) and during the 18th century, 10 more elements have been ...

  2. 40 CFR 264.1055 - Standards: Sampling connection systems.

    Science.gov (United States)

    2010-07-01

    ... operated to capture and transport all the purged process fluid to a waste management unit that complies... 40 Protection of Environment 25 2010-07-01 2010-07-01 false Standards: Sampling connection systems. 264.1055 Section 264.1055 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) SOLID...

  3. 40 CFR 264.92 - Ground-water protection standard.

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 25 2010-07-01 2010-07-01 false Ground-water protection standard. 264... Releases From Solid Waste Management Units § 264.92 Ground-water protection standard. The owner or operator... establish this ground-water protection standard in the facility permit when hazardous constituents have been...

  4. 27 CFR 26.264 - Determination of tax on wine.

    Science.gov (United States)

    2010-04-01

    ... 27 Alcohol, Tobacco Products and Firearms 1 2010-04-01 2010-04-01 false Determination of tax on wine. 26.264 Section 26.264 Alcohol, Tobacco Products and Firearms ALCOHOL AND TOBACCO TAX AND TRADE BUREAU, DEPARTMENT OF THE TREASURY LIQUORS LIQUORS AND ARTICLES FROM PUERTO RICO AND THE VIRGIN ISLANDS Procedure at Port of Entry From the Virgin...

  5. 40 CFR 264.1083 - Waste determination procedures.

    Science.gov (United States)

    2010-07-01

    ... hazardous waste stream is placed in a waste management unit exempted under the provisions of § 264.1082(c)(1... waste stream is placed in the exempt waste management unit, and thereafter update the information used... waste management unit exempted under the provisions of § 264.1082(c)(1) of this subpart from using air...

  6. 40 CFR 264.173 - Management of containers.

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 25 2010-07-01 2010-07-01 false Management of containers. 264.173... Use and Management of Containers § 264.173 Management of containers. (a) A container holding hazardous... container holding hazardous waste must not be opened, handled, or stored in a manner which may rupture the...

  7. 40 CFR 264.171 - Condition of containers.

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 25 2010-07-01 2010-07-01 false Condition of containers. 264.171... Use and Management of Containers § 264.171 Condition of containers. If a container holding hazardous... leak, the owner or operator must transfer the hazardous waste from this container to a container that...

  8. 40 CFR 264.35 - Required aisle space.

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 25 2010-07-01 2010-07-01 false Required aisle space. 264.35 Section... Preparedness and Prevention § 264.35 Required aisle space. The owner or operator must maintain aisle space to... Regional Administrator that aisle space is not needed for any of these purposes. [Comment: Part 270 of this...

  9. Evidence for lipopolysaccharide-induced differentiation of RAW264 ...

    Indian Academy of Sciences (India)

    Unknown

    Dose-dependent morphological transformation of. RAW264⋅7 cells by LPS. RAW264⋅7 cells were cultured on glass slides in a slide chamber in the presence of various given doses of LPS. After two days, slides were fixed and stained as described in § 2. Slides were observed under a microscope with a digital camera and ...

  10. The H.264/MPEG4 advanced video coding

    Science.gov (United States)

    Gromek, Artur

    2009-06-01

    H.264/MPEG4-AVC is the newest video coding standard recommended by International Telecommunication Union - Telecommunication Standardization Section (ITU-T) and the ISO/IEC Moving Picture Expert Group (MPEG). The H.264/MPEG4-AVC has recently become leading standard for generic audiovisual services, since deployment for digital television. Nowadays is commonly used in wide range of video application ranging like mobile services, videoconferencing, IPTV, HDTV, video storage and many more. In this article, author briefly describes the technology applied in the H.264/MPEG4-AVC video coding standard, the way of real-time implementation and the way of future development.

  11. Improved Intra-coding Methods for H.264/AVC

    Directory of Open Access Journals (Sweden)

    Li Song

    2009-01-01

    Full Text Available The H.264/AVC design adopts a multidirectional spatial prediction model to reduce spatial redundancy, where neighboring pixels are used as a prediction for the samples in a data block to be encoded. In this paper, a recursive prediction scheme and an enhanced (block-matching algorithm BMA prediction scheme are designed and integrated into the state-of-the-art H.264/AVC framework to provide a new intra coding model. Extensive experiments demonstrate that the coding efficiency can be on average increased by 0.27 dB with comparison to the performance of the conventional H.264 coding model.

  12. AVC/H.264 patent portfolio license

    Science.gov (United States)

    Skandalis, Dean A.

    2006-08-01

    MPEG LA, LLC offers a joint patent license for the AVC (a/k/a H.264) Standard (ISO/IEC IS 14496-10:2004). Like MPEG LA's other licenses, the AVC Patent Portfolio License is offered for the convenience of the marketplace as an alternative enabling users to access essential intellectual property owned by many patent holders under a single license rather than negotiating licenses with each of them individually. The AVC Patent Portfolio License includes essential patents owned by DAEWOO Electronics Corporation; Electronics and Telecommunications Research Institute (ETRI); France Telecom, societe anonyme; Fujitsu Limited; Hitachi, Ltd.; Koninklijke Philips Electronics N.V.; LG Electronics Inc.; Matsushita Electric Industrial Co., Ltd.; Microsoft Corporation; Mitsubishi Electric Corporation; Robert Bosch GmbH; Samsung Electronics Co., Ltd.; Sedna Patent Services, LLC; Sharp Kabushiki Kaisha; Siemens AG; Sony Corporation; The Trustees of Columbia University in the City of New York; Toshiba Corporation; UB Video Inc.; and Victor Company of Japan, Limited. Another is expected also to join as of August 1, 2006. MPEG LA's objective is to provide worldwide access to as much AVC essential intellectual property as possible for the benefit of AVC users. Therefore, any party that believes it has essential patents is welcome to submit them for evaluation of their essentiality and inclusion in the License if found essential.

  13. AN EFFICIENT WATERMARKING SCHEME FOR H.264/AVC COMPRESSED VIDEO

    Directory of Open Access Journals (Sweden)

    Dawen Xu

    2014-08-01

    Full Text Available Since H.264/AVC is the most widely-deployed video coding standard and has gained dominance, the necessity of copyright protection and authentication that are appropriate for this standard is unquestionable. According to H.264/AVC specific codec architecture, an efficient watermarking scheme for H.264/AVC video is proposed. The watermark information is embedded into quantized residual coefficients by slightly modulating the coefficients with specific symbol encoding, instead of directly adding the watermark to the quantized coefficients. It is not necessary to fully decode H.264/AVC compressed stream both in the embedding and extracting processes. Experimental results show that the proposed scheme can preserve high imperceptibility while achieving enough robustness against various attacks such as requantization, transcoding, AWGN, brightness and contrast adjustment.

  14. 36 CFR 264.3 - Licensing for commercial use.

    Science.gov (United States)

    2010-07-01

    ... PROPERTY MANAGEMENT Official Forest Service Insignia § 264.3 Licensing for commercial use. (a) Each... definite expiration date shall be specified. (3) The license shall be nonexclusive. (4) Licensees are not...

  15. High Definition Video Streaming Using H.264 Video Compression

    OpenAIRE

    Bechqito, Yassine

    2009-01-01

    This thesis presents high definition video streaming using H.264 codec implementation. The experiment carried out in this study was done for an offline streaming video but a model for live high definition streaming is introduced, as well. Prior to the actual experiment, this study describes digital media streaming. Also, the different technologies involved in video streaming are covered. These include streaming architecture and a brief overview on H.264 codec as well as high definition t...

  16. Video Compression Study: h.265 vs h.264

    Science.gov (United States)

    Pryor, Jonathan

    2016-01-01

    H.265 video compression (also known as High Efficiency Video Encoding (HEVC)) promises to provide double the video quality at half the bandwidth, or the same quality at half the bandwidth of h.264 video compression [1]. This study uses a Tektronix PQA500 to determine the video quality gains by using h.265 encoding. This study also compares two video encoders to see how different implementations of h.264 and h.265 impact video quality at various bandwidths.

  17. 40 CFR 264.97 - General ground-water monitoring requirements.

    Science.gov (United States)

    2010-07-01

    ... requirements. 264.97 Section 264.97 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) SOLID... FACILITIES Releases From Solid Waste Management Units § 264.97 General ground-water monitoring requirements... statistical procedures to comply with § 264.97(i)(5), the pql must be proposed by the owner or operator and...

  18. Improved lossless intra coding for H.264/MPEG-4 AVC.

    Science.gov (United States)

    Lee, Yung-Lyul; Han, Ki-Hun; Sullivan, Gary J

    2006-09-01

    A new lossless intra coding method based on sample-by-sample differential pulse code modulation (DPCM) is presented as an enhancement of the H.264/MPEG-4 AVC standard. The H.264/AVC design includes a multidirectional spatial prediction method to reduce spatial redundancy by using neighboring samples as a prediction for the samples in a block of data to be encoded. In the new lossless intra coding method, the spatial prediction is performed based on samplewise DPCM instead of in the block-based manner used in the current H.264/AVC standard, while the block structure is retained for the residual difference entropy coding process. We show that the new method, based on samplewise DPCM, does not have a major complexity penalty, despite its apparent pipeline dependencies. Experiments show that the new lossless intra coding method reduces the bit rate by approximately 12% in comparison with the lossless intra coding method previously included in the H.264/AVC standard. As a result, the new method is currently being adopted into the H.264/AVC standard in a new enhancement project.

  19. 40 CFR 264.150 - State assumption of responsibility.

    Science.gov (United States)

    2010-07-01

    ... FACILITIES Financial Requirements § 264.150 State assumption of responsibility. (a) If a State either assumes legal responsibility for an owner's or operator's compliance with the closure, post-closure care, or... 40 Protection of Environment 25 2010-07-01 2010-07-01 false State assumption of responsibility...

  20. 40 CFR 264.1063 - Test methods and procedures.

    Science.gov (United States)

    2010-07-01

    ... CFR part 60. (2) The detection instrument shall meet the performance criteria of Reference Method 21... determined as set forth in Reference Method 21. (3) The instrument probe shall be traversed around all... 40 Protection of Environment 25 2010-07-01 2010-07-01 false Test methods and procedures. 264.1063...

  1. Evidence for lipopolysaccharide-induced differentiation of RAW264 ...

    Indian Academy of Sciences (India)

    Flow cytometric studies indicate that the cell surface markers known to be expressed on dendritic cells and involved in antigen presentation and T cell activation (B7.1, B7.2, CD40, MHC class II antigens and CD1d) were also markedly upregulated on LPS-treated RAW264.7 cells. Our results suggest the possibility that LPS ...

  2. Adaptive deblocking and deringing of H.264/AVC video sequences

    DEFF Research Database (Denmark)

    Nadernejad, Ehsan; Burini, Nino; Forchhammer, Søren

    2013-01-01

    We present a method to reduce blocking and ringing artifacts in H.264/AVC video sequences. For deblocking, the proposed method uses a quality measure of a block based coded image to find filtering modes. Based on filtering modes, the images are segmented to three classes and a specific deblocking...

  3. 40 CFR 264.315 - Special requirements for containers.

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 25 2010-07-01 2010-07-01 false Special requirements for containers... FACILITIES Landfills § 264.315 Special requirements for containers. Unless they are very small, such as an ampule, containers must be either: (a) At least 90 percent full when placed in the landfill; or (b...

  4. 8 CFR 264.1 - Registration and fingerprinting.

    Science.gov (United States)

    2010-01-01

    ... United States under section 212(d)(5) of the Immigration and Nationality Act; aliens whose claimed entry... AND FINGERPRINTING OF ALIENS IN THE UNITED STATES § 264.1 Registration and fingerprinting. (a...—Aliens admitted as nonimmigrants; aliens paroled into the United States under section 212(d)(5) of the...

  5. 40 CFR 264.221 - Design and operating requirements.

    Science.gov (United States)

    2010-07-01

    ... render the wastes hazardous for reasons other than the toxicity characteristic in § 261.24 of this... waste from migrating beyond the liner to adjacent subsurface soil, ground water, or surface water at any....221 Section 264.221 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) SOLID WASTES...

  6. 40 CFR 264.34 - Access to communications or alarm system.

    Science.gov (United States)

    2010-07-01

    ... FACILITIES Preparedness and Prevention § 264.34 Access to communications or alarm system. (a) Whenever... 40 Protection of Environment 25 2010-07-01 2010-07-01 false Access to communications or alarm system. 264.34 Section 264.34 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) SOLID...

  7. Efficient reversible data hiding in encrypted H.264/AVC videos

    Science.gov (United States)

    Xu, Dawen; Wang, Rangding

    2014-09-01

    Due to the security and privacy-preserving requirements for cloud data management, it is sometimes desired that video content is accessible in an encrypted form. Reversible data hiding in the encrypted domain is an emerging technology, as it can perform data hiding in encrypted videos without decryption, which preserves the confidentiality of the content. Furthermore, the original cover can be losslessly restored after decryption and data extraction. An efficient reversible data hiding scheme for encrypted H.264/AVC videos is proposed. During H.264/AVC encoding, the intraprediction mode, motion vector difference, and the sign bits of the residue coefficients are encrypted using a standard stream cipher. Then, the data-hider who does not know the original video content, may reversibly embed secret data into the encrypted H.264/AVC video by using a modified version of the histogram shifting technique. A scale factor is utilized for selecting the embedding zone, which is scalable for different capacity requirements. With an encrypted video containing hidden data, data extraction can be carried out either in the encrypted or decrypted domain. In addition, real reversibility is realized so that data extraction and video recovery are free of any error. Experimental results demonstrate the feasibility and efficiency of the proposed scheme.

  8. Entropy coders of the H.264/AVC standard

    CERN Document Server

    Tian, Xiaohua; Lian, Yong

    2010-01-01

    This book presents a collection of algorithms and VLSI architectures of entropy (or statistical) codecs of recent video compression standards, with focus on the H.264/AVC standard. For any visual data compression scheme, there exists a combination of two, or all of the following three stages: spatial, temporal, and statistical compression. General readers are first introduced with the various algorithms of the statistical coders. The VLSI implementations are also reviewed and discussed. Readers with limited hardware design background are also introduced with a design methodology starting from

  9. 27 CFR 44.264a - To a foreign-trade zone.

    Science.gov (United States)

    2010-04-01

    ... 27 Alcohol, Tobacco Products and Firearms 2 2010-04-01 2010-04-01 false To a foreign-trade zone. 44.264a Section 44.264a Alcohol, Tobacco Products and Firearms ALCOHOL AND TOBACCO TAX AND TRADE... Warehouses Notice of Removal of Shipment § 44.264a To a foreign-trade zone. Where cigars are withdrawn from a...

  10. pB264, a small, mobilizable, temperature sensitive plasmid from Rhodococcus

    Science.gov (United States)

    Lessard, Philip A; O'Brien, Xian M; Currie, Devin H; Sinskey, Anthony J

    2004-01-01

    Background Gram-positive bacteria of the genus Rhodococcus have shown an extraordinary capacity for metabolizing recalcitrant organic compounds. One hindrance to the full exploitation of Rhodococcus is the dearth of genetic tools available for strain manipulation. To address this issue, we sought to develop a plasmid-based system for genetic manipulation of a variety of Rhodococcus strains. Results We isolated and sequenced pB264, a 4,970 bp cryptic plasmid from Rhodococcus sp. B264-1 with features of a theta-type replication mechanism. pB264 was nearly identical to pKA22, a previously sequenced but uncharacterized cryptic plasmid. Derivatives of pB264 replicate in a diverse range of Rhodococcus species, showing that this plasmid does not bear the same host range restrictions that have been exhibited by other theta replicating plasmids. Replication or maintenance of pB264 is inhibited at 37°C, making pB264 useful as a suicide vector for genetic manipulation of Rhodococcus. A series of deletions revealed that ca. 1.3 kb from pB264 was sufficient to support replication and stable inheritance of the plasmid. This region includes two open reading frames that encode functions (RepAB) that can support replication of pB264 derivatives in trans. Rhodococcus sp. B264-1 will mobilize pB264 into other Rhodococcus species via conjugation, making it possible to genetically modify bacterial strains that are otherwise difficult to transform. The cis-acting element (oriT) required for conjugal transfer of pB264 resides within a ca. 0.7 kb region that is distinct from the regions responsible for replication. Conclusion Shuttle vectors derived from pB264 will be useful for genetic studies and strain improvement in Rhodococcus, and will also be useful for studying the processes of theta replication and conjugal transfer among actinomycetes. PMID:15084226

  11. 40 CFR 264.555 - Disposal of CAMU-eligible wastes in permitted hazardous waste landfills.

    Science.gov (United States)

    2010-07-01

    ... permitted hazardous waste landfills. 264.555 Section 264.555 Protection of Environment ENVIRONMENTAL...-eligible wastes in permitted hazardous waste landfills. (a) The Regional Administrator with regulatory... hazardous waste landfills not located at the site from which the waste originated, without the wastes...

  12. 40 CFR 264.114 - Disposal or decontamination of equipment, structures and soils.

    Science.gov (United States)

    2010-07-01

    ... equipment, structures and soils. 264.114 Section 264.114 Protection of Environment ENVIRONMENTAL PROTECTION... of equipment, structures and soils. During the partial and final closure periods, all contaminated equipment, structures and soils must be properly disposed of or decontaminated unless otherwise specified in...

  13. Temporal scalability comparison of the H.264/SVC and distributed video codec

    DEFF Research Database (Denmark)

    Huang, Xin; Ukhanova, Ann; Belyaev, Evgeny

    2009-01-01

    The problem of the multimedia scalable video streaming is a current topic of interest. There exist many methods for scalable video coding. This paper is focused on the scalable extension of H.264/AVC (H.264/SVC) and distributed video coding (DVC). The paper presents an efficiency comparison of SVC...

  14. 40 CFR 264.1054 - Standards: Pressure relief devices in gas/vapor service.

    Science.gov (United States)

    2010-07-01

    ... gas/vapor service. 264.1054 Section 264.1054 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY... relief devices in gas/vapor service. (a) Except during pressure releases, each pressure relief device in gas/vapor service shall be operated with no detectable emissions, as indicated by an instrument...

  15. 8 CFR 264.2 - Application for creation of record of permanent residence.

    Science.gov (United States)

    2010-01-01

    ... permanent residence. 264.2 Section 264.2 Aliens and Nationality DEPARTMENT OF HOMELAND SECURITY IMMIGRATION... record of permanent residence. (a) Jurisdiction. An applicant who believes that he/she is eligible for presumption of lawful admission for permanent residence under § 101.1 or § 101.2 of this chapter or for lawful...

  16. 40 CFR 264.551 - Grandfathered Corrective Action Management Units (CAMUs).

    Science.gov (United States)

    2010-07-01

    ...; (E) Hydrological and other relevant environmental conditions at the facility which may influence the migration of any potential or actual releases; and (F) Potential for exposure of humans and environmental... Management Units (CAMUs). 264.551 Section 264.551 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY...

  17. 40 CFR 264.149 - Use of State-required mechanisms.

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 25 2010-07-01 2010-07-01 false Use of State-required mechanisms. 264... Financial Requirements § 264.149 Use of State-required mechanisms. (a) For a facility located in a State... coverage, an owner or operator may use State-required financial mechanisms to meet the requirements of...

  18. 40 CFR 264.145 - Financial assurance for post-closure care.

    Science.gov (United States)

    2010-07-01

    ... care. 264.145 Section 264.145 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED... by an amount equivalent to 85 percent of the most recent investment rate or of the equivalent coupon... connection with that procedure, no matters came to his attention which caused him to believe that the...

  19. Content Adaptive True Motion Estimator for H.264 Video Compression

    Directory of Open Access Journals (Sweden)

    P. Kulla

    2007-12-01

    Full Text Available Content adaptive true motion estimator for H.264 video coding is a fast block-based matching estimator with implemented multi-stage approach to estimate motion fields between two image frames. It considers the theory of 3D scene objects projection into 2D image plane for selection of motion vector candidates from the higher stages. The stages of the algorithm and its hierarchy are defined upon motion estimation reliability measurement (image blocks including two different directions of spatial gradient, blocks with one dominant spatial gradient and blocks including minimal spatial gradient. Parameters of the image classification into stages are set adaptively upon image structure. Due to search strategy are the estimated motion fields more corresponding to a true motion in an image sequence as in the case of conventional motion estimation algorithms that use fixed sets of motion vector candidates from tight neighborhood.

  20. Fast prototyping H.264 deblocking filter using ESL tools

    International Nuclear Information System (INIS)

    Damak, T.; Werda, I.; Masmoud, N.; Bilavarn, S.

    2011-01-01

    This paper presents a design methodology for hardware/software (HW/SW) architecture design using ESL tools (Electronic System Level). From C++ descriptions, our design flow is able to generate hardware blocks running with a software part and all necessary codes to prototype the HW/SW system on Xilinx FPGAs. Therefore we use assistance of high level synthesis tools (Catapult C Synthesis), logic synthesis and Xilinx tools. As application, we developed an optimized Deblocking filter C code, designed to be used as a part of a complete H.264 video coding system [1]. Based on this code, we explored many configurations of Catapult Synthesis to analyze different area/time tradeoffs. Results show execution speedups of 95,5 pour cent compared to pure software execution etc.

  1. H.264 SVC Complexity Reduction Based on Likelihood Mode Decision

    Directory of Open Access Journals (Sweden)

    L. Balaji

    2015-01-01

    Full Text Available H.264 Advanced Video Coding (AVC was prolonged to Scalable Video Coding (SVC. SVC executes in different electronics gadgets such as personal computer, HDTV, SDTV, IPTV, and full-HDTV in which user demands various scaling of the same content. The various scaling is resolution, frame rate, quality, heterogeneous networks, bandwidth, and so forth. Scaling consumes more encoding time and computational complexity during mode selection. In this paper, to reduce encoding time and computational complexity, a fast mode decision algorithm based on likelihood mode decision (LMD is proposed. LMD is evaluated in both temporal and spatial scaling. From the results, we conclude that LMD performs well, when compared to the previous fast mode decision algorithms. The comparison parameters are time, PSNR, and bit rate. LMD achieve time saving of 66.65% with 0.05% detriment in PSNR and 0.17% increment in bit rate compared with the full search method.

  2. H.264 SVC Complexity Reduction Based on Likelihood Mode Decision.

    Science.gov (United States)

    Balaji, L; Thyagharajan, K K

    2015-01-01

    H.264 Advanced Video Coding (AVC) was prolonged to Scalable Video Coding (SVC). SVC executes in different electronics gadgets such as personal computer, HDTV, SDTV, IPTV, and full-HDTV in which user demands various scaling of the same content. The various scaling is resolution, frame rate, quality, heterogeneous networks, bandwidth, and so forth. Scaling consumes more encoding time and computational complexity during mode selection. In this paper, to reduce encoding time and computational complexity, a fast mode decision algorithm based on likelihood mode decision (LMD) is proposed. LMD is evaluated in both temporal and spatial scaling. From the results, we conclude that LMD performs well, when compared to the previous fast mode decision algorithms. The comparison parameters are time, PSNR, and bit rate. LMD achieve time saving of 66.65% with 0.05% detriment in PSNR and 0.17% increment in bit rate compared with the full search method.

  3. Fast H.264/AVC FRExt intra coding using belief propagation.

    Science.gov (United States)

    Milani, Simone

    2011-01-01

    In the H.264/AVC FRExt coder, the coding performance of Intra coding significantly overcomes the previous still image coding standards, like JPEG2000, thanks to a massive use of spatial prediction. Unfortunately, the adoption of an extensive set of predictors induces a significant increase of the computational complexity required by the rate-distortion optimization routine. The paper presents a complexity reduction strategy that aims at reducing the computational load of the Intra coding with a small loss in the compression performance. The proposed algorithm relies on selecting a reduced set of prediction modes according to their probabilities, which are estimated adopting a belief-propagation procedure. Experimental results show that the proposed method permits saving up to 60 % of the coding time required by an exhaustive rate-distortion optimization method with a negligible loss in performance. Moreover, it permits an accurate control of the computational complexity unlike other methods where the computational complexity depends upon the coded sequence.

  4. The uptake of tocopherols by RAW 264.7 macrophages

    Directory of Open Access Journals (Sweden)

    Papas Andreas M

    2002-10-01

    Full Text Available Abstract Background Alpha-Tocopherol and gamma-tocopherol are the two major forms of vitamin E in human plasma and the primary lipid soluble antioxidants. The dietary intake of gamma-tocopherol is generally higher than that of alpha-tocopherol. However, alpha-tocopherol plasma levels are about four fold higher than those of gamma-tocopherol. Among other factors, a preferential cellular uptake of gamma-tocopherol over alpha-tocopherol could contribute to the observed higher plasma alpha-tocopherol levels. In this investigation, we studied the uptake and depletion of both alpha-tocopherol and gamma-tocopherol (separately and together in cultured RAW 264.7 macrophages. Similar studies were performed with alpha-tocopheryl quinone and gamma-tocopheryl quinone, which are oxidation products of tocopherols. Results RAW 264.7 macrophages showed a greater uptake of gamma-tocopherol compared to alpha-tocopherol (with uptake being defined as the net difference between tocopherol transported into the cells and loss due to catabolism and/or in vitro oxidation. Surprisingly, we also found that the presence of gamma-tocopherol promoted the cellular uptake of alpha-tocopherol. Mass balance considerations suggest that products other than quinone were formed during the incubation of tocopherols with macrophages. Conclusion Our data suggests that gamma-tocopherol could play a significant role in modulating intracellular antioxidant defence mechanisms. Moreover, we found the presence of gamma-tocopherol dramatically influenced the cellular accumulation of alpha-tocopherol, i.e., gamma-tocopherol promoted the accumulation of alpha-tocopherol. If these results could be extrapolated to in vivo conditions they suggest that gamma-tocopherol is selectively taken up by cells and removed from plasma more rapidly than alpha-tocopherol. This could, in part, contribute to the selective maintenance of alpha-tocopherol in plasma compared to gamma-tocopherol.

  5. Mode-dependent templates and scan order for H.264/AVC-based intra lossless coding.

    Science.gov (United States)

    Gu, Zhouye; Lin, Weisi; Lee, Bu-Sung; Lau, Chiew Tong; Sun, Ming-Ting

    2012-09-01

    In H.264/advanced video coding (AVC), lossless coding and lossy coding share the same entropy coding module. However, the entropy coders in the H.264/AVC standard were original designed for lossy video coding and do not yield adequate performance for lossless video coding. In this paper, we analyze the problem with the current lossless coding scheme and propose a mode-dependent template (MD-template) based method for intra lossless coding. By exploring the statistical redundancy of the prediction residual in the H.264/AVC intra prediction modes, more zero coefficients are generated. By designing a new scan order for each MD-template, the scanned coefficients sequence fits the H.264/AVC entropy coders better. A fast implementation algorithm is also designed. With little computation increase, experimental results confirm that the proposed fast algorithm achieves about 7.2% bit saving compared with the current H.264/AVC fidelity range extensions high profile.

  6. WebM as an alternative to H.264? : Investigation of the usage of open source software and open standards

    NARCIS (Netherlands)

    Staalduinen, M. van; Prins, M.J.

    2011-01-01

    WebM is a new multimedia format often postitioned as an open and free to use alternative to the industry standard H.264. H.264 is currently the most popular web-video format, used for broadcast video, VoD but also Catch-Up TV services. Comparisons between WebM's VP8 video format and the H.264 format

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  6. File list: His.Bld.05.AllAg.RAW_264PERIOD7 [Chip-atlas[Archive

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  7. pB264, a small, mobilizable, temperature sensitive plasmid from Rhodococcus

    Directory of Open Access Journals (Sweden)

    Currie Devin H

    2004-04-01

    Full Text Available Abstract Background Gram-positive bacteria of the genus Rhodococcus have shown an extraordinary capacity for metabolizing recalcitrant organic compounds. One hindrance to the full exploitation of Rhodococcus is the dearth of genetic tools available for strain manipulation. To address this issue, we sought to develop a plasmid-based system for genetic manipulation of a variety of Rhodococcus strains. Results We isolated and sequenced pB264, a 4,970 bp cryptic plasmid from Rhodococcus sp. B264-1 with features of a theta-type replication mechanism. pB264 was nearly identical to pKA22, a previously sequenced but uncharacterized cryptic plasmid. Derivatives of pB264 replicate in a diverse range of Rhodococcus species, showing that this plasmid does not bear the same host range restrictions that have been exhibited by other theta replicating plasmids. Replication or maintenance of pB264 is inhibited at 37°C, making pB264 useful as a suicide vector for genetic manipulation of Rhodococcus. A series of deletions revealed that ca. 1.3 kb from pB264 was sufficient to support replication and stable inheritance of the plasmid. This region includes two open reading frames that encode functions (RepAB that can support replication of pB264 derivatives in trans. Rhodococcus sp. B264-1 will mobilize pB264 into other Rhodococcus species via conjugation, making it possible to genetically modify bacterial strains that are otherwise difficult to transform. The cis-acting element (oriT required for conjugal transfer of pB264 resides within a ca. 0.7 kb region that is distinct from the regions responsible for replication. Conclusion Shuttle vectors derived from pB264 will be useful for genetic studies and strain improvement in Rhodococcus, and will also be useful for studying the processes of theta replication and conjugal transfer among actinomycetes.

  8. Scalable Fast Rate-Distortion Optimization for H.264/AVC

    Directory of Open Access Journals (Sweden)

    Yu Hongtao

    2006-01-01

    Full Text Available The latest H.264/AVC video coding standard aims at significantly improving compression performance compared to all existing video coding standards. In order to achieve this, variable block-size inter- and intra-coding, with block sizes as large as and as small as , is used to enable very precise depiction of motion and texture details. The Lagrangian rate-distortion optimization (RDO can be employed to select the best coding mode. However, exhaustively searching through all coding modes is computationally expensive. This paper proposes a scalable fast RDO algorithm to effectively choose the best coding mode without exhaustively searching through all the coding modes. The statistical properties of MBs are analyzed to determine the order of coding modes in the mode decision priority queue such that the most probable mode will be checked first, followed by the second most probable mode, and so forth. The process will be terminated as soon as the computed rate-distortion (RD cost is below a threshold which is content adaptive and is also dependent on the RD cost of the previous MBs. By adjusting the threshold we can choose a good tradeoff between timesaving and peak signal-to-noise (PSNR ratio. Experimental results show that the proposed fast RDO algorithm can drastically reduce the encoding time up to 50% with negligible loss of coding efficiency.

  9. Zero-block mode decision algorithm for H.264/AVC.

    Science.gov (United States)

    Lee, Yu-Ming; Lin, Yinyi

    2009-03-01

    In the previous paper , we proposed a zero-block intermode decision algorithm for H.264 video coding based upon the number of zero-blocks of 4 x 4 DCT coefficients between the current macroblock and the co-located macroblock. The proposed algorithm can achieve significant improvement in computation, but the computation performance is limited for high bit-rate coding. To improve computation efficiency, in this paper, we suggest an enhanced zero-block decision algorithm, which uses an early zero-block detection method to compute the number of zero-blocks instead of direct DCT and quantization (DCT/Q) calculation and incorporates two adequate decision methods into semi-stationary and nonstationary regions of a video sequence. In addition, the zero-block decision algorithm is also applied to the intramode prediction in the P frame. The enhanced zero-block decision algorithm brings out a reduction of average 27% of total encoding time compared to the zero-block decision algorithm.

  10. 26 CFR 1.264-4 - Other life insurance, endowment, or annuity contracts.

    Science.gov (United States)

    2010-04-01

    ... (CONTINUED) INCOME TAX (CONTINUED) INCOME TAXES Items Not Deductible § 1.264-4 Other life insurance... insurer, insured, or any other person (other than by gift, bequest, or inheritance, or in a transaction to...

  11. Arithmetic coding with adaptive context-tree weighting for the H.264 video coders

    Science.gov (United States)

    Hong, Danny; van der Schaar, Mihaela; Pesquet-Popescu, Beatrice

    2004-01-01

    We propose applying an adaptive context-tree weighting (CTW) method in the H.264 video coders. We first investigate two different ways to incorporating the CTW method into an H.264 coder and compare the coding effectiveness of using the method with that of using the context models specified in the H.264 standard. We then describe a novel approach for automatically adapting the CTW method based on the syntactic element to be coded and the encoding parameters. We show that our CTW-based arithmetic coding method yields similar or better compression results compared with the context-based adaptive arithmetic coding method used in H.264, without having to specify so many context models.

  12. Upregulating Nonneuronal Cholinergic Activity Decreases TNF Release from Lipopolysaccharide-Stimulated RAW264.7 Cells

    Directory of Open Access Journals (Sweden)

    Yi Lv

    2014-01-01

    Full Text Available Nonneuronal cholinergic system plays a primary role in maintaining homeostasis. It has been proved that endogenous neuronal acetylcholine (ACh could play an anti-inflammatory role, and exogenous cholinergic agonists could weaken macrophages inflammatory response to lipopolysaccharide (LPS stimulation through activation of α7 subunit-containing nicotinic acetylcholine receptor (α7nAChR. We assumed that nonneuronal cholinergic system existing in macrophages could modulate inflammation through autocrine ACh and expressed α7nAChR on the cells. Therefore, we explored whether LPS continuous stimulation could upregulate the nonneuronal cholinergic activity in macrophages and whether increasing autocrine ACh could decrease TNF release from the macrophages. The results showed that, in RAW264.7 cells incubated with LPS for 20 hours, the secretion of ACh was significantly decreased at 4 h and then gradually increased, accompanied with the enhancement of α7nAChR expression level. The release of TNF was greatly increased from RAW264.7 cells at 4 h and 8 h exposure to LPS; however, it was suppressed at 20 h. Upregulating choline acetyltransferase (ChAT expression through ChAT gene transfection could enhance ACh secretion and reduce TNF release from the infected RAW264. 7cells. The results indicated that LPS stimulation could modulate the activity of nonneuronal cholinergic system of RAW264.7 cells. Enhancing autocrine ACh production could attenuate TNF release from RAW264.7 cells.

  13. Endoplasmic Reticulum Stress Cooperates in Zearalenone-Induced Cell Death of RAW 264.7 Macrophages

    Directory of Open Access Journals (Sweden)

    Fenglei Chen

    2015-08-01

    Full Text Available Zearalenone (ZEA is a fungal mycotoxin that causes cell apoptosis and necrosis. However, little is known about the molecular mechanisms of ZEA toxicity. The objective of this study was to explore the effects of ZEA on the proliferation and apoptosis of RAW 264.7 macrophages and to uncover the signaling pathway underlying the cytotoxicity of ZEA in RAW 264.7 macrophages. This study demonstrates that the endoplasmic reticulum (ER stress pathway cooperated in ZEA-induced cell death of the RAW 264.7 macrophages. Our results show that ZEA treatment reduced the viability of RAW 264.7 macrophages in a dose- and time-dependent manner as shown by the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide assay (MTT and flow cytometry assay. Western blots analysis revealed that ZEA increased the expression of glucose-regulated protein 78 (GRP78 and CCAAT/enhancer binding protein homologous protein (CHOP, two ER stress-related marker genes. Furthermore, treating the cells with the ER stress inhibitors 4-phenylbutyrate (4-PBA or knocking down CHOP, using lentivirus encoded short hairpin interfering RNAs (shRNAs, significantly diminished the ZEA-induced increases in GRP78 and CHOP, and cell death. In summary, our results suggest that ZEA induces the apoptosis and necrosis of RAW 264.7 macrophages in a dose- and time-dependent manner via the ER stress pathway in which the activation of CHOP plays a critical role.

  14. Intra-Skip in Inter-Frame Coding of H.264/AVC

    Directory of Open Access Journals (Sweden)

    Hui Su

    2009-01-01

    Full Text Available In inter-frame coding of H.264/AVC standard, not only seven inter-partition modes but also intra-modes are taken into account for seeking the best coding mode so as to maintain higher encoding efficiency by sacrificing the speed of H.264/AVC encoder. Aiming at intra-skip, this paper proposes a novel mathematical model for intra-skip in inter-frame coding to alleviate the complexity of the process; the model provides remarkable performance increment by cutting down encoding time while accompanying very minor bitrate increase. The critical advantage of this proposed scheme most emphasized on is that it can optimize H.264 encoder in conjunction with any proposed fast inter- and intra-methods which are focusing on inter-partition mode decision, motion search algorithms, and fast intra-algorithms.

  15. ADAPTIVE GOP STRUCTURE TO H.264/AVC BASED ON SCENE CHANGE

    Directory of Open Access Journals (Sweden)

    S. Sowmyayani

    2014-08-01

    Full Text Available This paper proposes an adaptive GOP structure with the new logic of frame comparison in H.264/AVC to achieve better quality and reduce bit rate. Initially Group of Pictures (GOP is set to a fixed size. Frames are compared within that GOP using correlation. According to the correlation, GOP is changed within that fixed size. So, there will be no GOP size greater than that fixed size. This method does not calculate any threshold. Hence the time needed to calculate global or local threshold is eliminated. It is integrated with conventional video codec H.264/AVC. This method is compared with H.264/AVC of fixed GOP structure of sizes 4, 8, 12, 16, 32 and GOP structure with the length of entire video. The proposed method achieved gain in bit rate from 0.49% to 69.75% and PSNR gain from 2.5% to 0.3%.

  16. Optimized H.264/AVC-Based Bit Stream Switching for Mobile Video Streaming

    Directory of Open Access Journals (Sweden)

    Liebl Günther

    2006-01-01

    Full Text Available We show the suitability of H.264/MPEG-4 AVC extended profile for wireless video streaming applications. In particular, we exploit the advanced bit stream switching capabilities using SP/SI pictures defined in the H.264/MPEG-4 AVC standard. For both types of switching pictures, optimized encoders are developed. We introduce a framework for dynamic switching and frame scheduling. For this purpose we define an appropriate abstract representation for media encoded for video streaming, as well as for the characteristics of wireless variable bit rate channels. The achievable performance gains over H.264/MPEG-4 AVC with constant bit rate (CBR encoding are shown for wireless video streaming over enhanced GPRS (EGPRS.

  17. Segmentation-driven compound document coding based on H.264/AVC-INTRA.

    Science.gov (United States)

    Zaghetto, Alexandre; de Queiroz, Ricardo L

    2007-07-01

    In this paper, we explore H.264/AVC operating in intraframe mode to compress a mixed image, i.e., composed of text, graphics, and pictures. Even though mixed contents (compound) documents usually require the use of multiple compressors, we apply a single compressor for both text and pictures. For that, distortion is taken into account differently between text and picture regions. Our approach is to use a segmentation-driven adaptation strategy to change the H.264/AVC quantization parameter on a macroblock by macroblock basis, i.e., we deviate bits from pictorial regions to text in order to keep text edges sharp. We show results of a segmentation driven quantizer adaptation method applied to compress documents. Our reconstructed images have better text sharpness compared to straight unadapted coding, at negligible visual losses on pictorial regions. Our results also highlight the fact that H.264/AVC-INTRA outperforms coders such as JPEG-2000 as a single coder for compound images.

  18. Evaluation of H.264 and H.265 full motion video encoding for small UAS platforms

    Science.gov (United States)

    McGuinness, Christopher D.; Walker, David; Taylor, Clark; Hill, Kerry; Hoffman, Marc

    2016-05-01

    Of all the steps in the image acquisition and formation pipeline, compression is the only process that degrades image quality. A selected compression algorithm succeeds or fails to provide sufficient quality at the requested compression rate depending on how well the algorithm is suited to the input data. Applying an algorithm designed for one type of data to a different type often results in poor compression performance. This is mostly the case when comparing the performance of H.264, designed for standard definition data, to HEVC (High Efficiency Video Coding), which the Joint Collaborative Team on Video Coding (JCT-VC) designed for high-definition data. This study focuses on evaluating how HEVC compares to H.264 when compressing data from small UAS platforms. To compare the standards directly, we assess two open-source traditional software solutions: x264 and x265. These software-only comparisons allow us to establish a baseline of how much improvement can generally be expected of HEVC over H.264. Then, specific solutions leveraging different types of hardware are selected to understand the limitations of commercial-off-the-shelf (COTS) options. Algorithmically, regardless of the implementation, HEVC is found to provide similar quality video as H.264 at 40% lower data rates for video resolutions greater than 1280x720, roughly 1 Megapixel (MPx). For resolutions less than 1MPx, H.264 is an adequate solution though a small (roughly 20%) compression boost is earned by employing HEVC. New low cost, size, weight, and power (CSWAP) HEVC implementations are being developed and will be ideal for small UAS systems.

  19. Residual DPCM about Motion Compensated Residual Signal for H.264 Lossless Coding

    Science.gov (United States)

    Han, Ki-Hun; Rao, Kamisetty R.; Lee, Yung-Lyul

    In this letter, a new Inter lossless coding method based on a residual DPCM (Differential Pulse Code Modulation) is proposed to improve compression ratio in the H.264 standard. Since the spatial correlation in a residual block can be further exploited among the residual signals after motion estimation/compensation, horizontal or vertical DPCM in the residual signals can be applied to further reduce the magnitudes of the residual signals. The proposed method reduces the average bitrates of 3.5% compared with the Inter lossless coding of the H.264 standard.

  20. 49 CFR 195.264 - Impoundment, protection against entry, normal/emergency venting or pressure/vacuum relief for...

    Science.gov (United States)

    2010-10-01

    .../emergency venting or pressure/vacuum relief for aboveground breakout tanks. 195.264 Section 195.264.../vacuum relief for aboveground breakout tanks. (a) A means must be provided for containing hazardous... relief venting must be provided for each atmospheric pressure breakout tank. Pressure/vacuum-relieving...

  1. 45 CFR 264.30 - What procedures exist to ensure cooperation with the child support enforcement requirements?

    Science.gov (United States)

    2010-10-01

    ... the child support enforcement requirements? 264.30 Section 264.30 Public Welfare Regulations Relating to Public Welfare OFFICE OF FAMILY ASSISTANCE (ASSISTANCE PROGRAMS), ADMINISTRATION FOR CHILDREN AND FAMILIES, DEPARTMENT OF HEALTH AND HUMAN SERVICES OTHER ACCOUNTABILITY PROVISIONS What Specific Rules Apply...

  2. Transcoding method from H.264/AVC to high efficiency video coding based on similarity of intraprediction, interprediction, and motion vector

    Science.gov (United States)

    Liu, Mei-Feng; Zhong, Guo-Yun; He, Xiao-Hai; Qing, Lin-Bo

    2016-09-01

    Currently, most video resources on line are encoded in the H.264/AVC format. More fluent video transmission can be obtained if these resources are encoded in the newest international video coding standard: high efficiency video coding (HEVC). In order to improve the video transmission and storage on line, a transcoding method from H.264/AVC to HEVC is proposed. In this transcoding algorithm, the coding information of intraprediction, interprediction, and motion vector (MV) in H.264/AVC video stream are used to accelerate the coding in HEVC. It is found through experiments that the region of interprediction in HEVC overlaps that in H.264/AVC. Therefore, the intraprediction for the region in HEVC, which is interpredicted in H.264/AVC, can be skipped to reduce coding complexity. Several macroblocks in H.264/AVC are combined into one PU in HEVC when the MV difference between two of the macroblocks in H.264/AVC is lower than a threshold. This method selects only one coding unit depth and one prediction unit (PU) mode to reduce the coding complexity. An MV interpolation method of combined PU in HEVC is proposed according to the areas and distances between the center of one macroblock in H.264/AVC and that of the PU in HEVC. The predicted MV accelerates the motion estimation for HEVC coding. The simulation results show that our proposed algorithm achieves significant coding time reduction with a little loss in bitrates distortion rate, compared to the existing transcoding algorithms and normal HEVC coding.

  3. 25 CFR 26.4 - Who administers the Job Placement and Training Program?

    Science.gov (United States)

    2010-04-01

    ... 25 Indians 1 2010-04-01 2010-04-01 false Who administers the Job Placement and Training Program... PLACEMENT AND TRAINING PROGRAM General Applicability § 26.4 Who administers the Job Placement and Training Program? The Job Placement and Training Program is administered by the Bureau of Indian Affairs or a...

  4. Power saving control for the mobile DVB-H receivers based on H.264/SVC standard

    DEFF Research Database (Denmark)

    Belyaev, Evgeny; Grinko, Vitaly; Ukhanova, Ann

    2009-01-01

    This paper discusses the utilization of scalable extension of H.264/AVC standard in digital video broadcasting for handheld devices. In this area the problem of mobile receiver power consumption is critically important. This paper amplifies the well-known idea of the time-slicing and allows...

  5. Fast compressed domain motion detection in H.264 video streams for video surveillance applications

    DEFF Research Database (Denmark)

    Szczerba, Krzysztof; Forchhammer, Søren; Støttrup-Andersen, Jesper

    2009-01-01

    This paper presents a novel approach to fast motion detection in H.264/MPEG-4 advanced video coding (AVC) compressed video streams for IP video surveillance systems. The goal is to develop algorithms which may be useful in a real-life industrial perspective by facilitating the processing of large...

  6. 40 CFR 264.101 - Corrective action for solid waste management units.

    Science.gov (United States)

    2010-07-01

    ..., AND DISPOSAL FACILITIES Releases From Solid Waste Management Units § 264.101 Corrective action for solid waste management units. (a) The owner or operator of a facility seeking a permit for the treatment... management unit at the facility, regardless of the time at which waste was placed in such unit. (b...

  7. VERITAS discovery of VHE emission from the FRI radio galaxy 3C 264

    Science.gov (United States)

    Mukherjee, Reshmi

    2018-03-01

    We report the VERITAS discovery of very-high-energy emission (VHE; > 100 GeV) from the FRI radio galaxy 3C 264, also known as NGC 3862. Nearly 12 hours of quality selected data, collected by VERITAS between 09 February 2018 and 16 March 2018 (UTC), were analyzed.

  8. No Reference Prediction of Quality Metrics for H.264 Compressed Infrared Image Sequences for UAV Applications

    DEFF Research Database (Denmark)

    Hossain, Kabir; Mantel, Claire; Forchhammer, Søren

    2018-01-01

    The framework for this research work is the acquisition of Infrared (IR) images from Unmanned Aerial Vehicles (UAV). In this paper we consider the No-Reference (NR) prediction of Full Reference Quality Metrics for Infrared (IR) video sequences which are compressed and thus distorted by an H.264...

  9. Inhibitory effect of emodin on raw 264.7 activated with double ...

    African Journals Online (AJOL)

    Inhibitory effect of emodin on raw 264.7 activated with double stranded RNA analogue poly I:C. Young-Jin Kim, Ji Young Lee, Hyun-Ju Kim, Do-Hoon Kim, Tae Hee Lee, Mi Suk Kang, You-Kyung Choi, Hye Lim Lee, Jaieun Kim, Hyo-Jin An, Wansu Park ...

  10. Bacterial lipopolysaccharide induces osteoclast formation in RAW 264.7 macrophage cells

    International Nuclear Information System (INIS)

    Islam, Shamima; Hassan, Ferdaus; Tumurkhuu, Gantsetseg; Dagvadorj, Jargalsaikhan; Koide, Naoki; Naiki, Yoshikazu; Mori, Isamu; Yoshida, Tomoaki; Yokochi, Takashi

    2007-01-01

    Lipopolysaccharide (LPS) is a potent bone resorbing factor. The effect of LPS on osteoclast formation was examined by using murine RAW 264.7 macrophage cells. LPS-induced the formation of multinucleated giant cells (MGC) in RAW 264.7 cells 3 days after the exposure. MGCs were positive for tartrate-resistant acid phosphatase (TRAP) activity. Further, MGC formed resorption pits on calcium-phosphate thin film that is a substrate for osteoclasts. Therefore, LPS was suggested to induce osteoclast formation in RAW 264.7 cells. LPS-induced osteoclast formation was abolished by anti-tumor necrosis factor (TNF)-α antibody, but not antibodies to macrophage-colony stimulating factor (M-CSF) and receptor activator of nuclear factor (NF)-κB ligand (RANKL). TNF-α might play a critical role in LPS-induced osteoclast formation in RAW 264.7 cells. Inhibitors of NF-κB and stress activated protein kinase (SAPK/JNK) prevented the LPS-induced osteoclast formation. The detailed mechanism of LPS-induced osteoclast formation is discussed

  11. FPSoC-Based Architecture for a Fast Motion Estimation Algorithm in H.264/AVC

    Directory of Open Access Journals (Sweden)

    Obianuju Ndili

    2009-01-01

    Full Text Available There is an increasing need for high quality video on low power, portable devices. Possible target applications range from entertainment and personal communications to security and health care. While H.264/AVC answers the need for high quality video at lower bit rates, it is significantly more complex than previous coding standards and thus results in greater power consumption in practical implementations. In particular, motion estimation (ME, in H.264/AVC consumes the largest power in an H.264/AVC encoder. It is therefore critical to speed-up integer ME in H.264/AVC via fast motion estimation (FME algorithms and hardware acceleration. In this paper, we present our hardware oriented modifications to a hybrid FME algorithm, our architecture based on the modified algorithm, and our implementation and prototype on a PowerPC-based Field Programmable System on Chip (FPSoC. Our results show that the modified hybrid FME algorithm on average, outperforms previous state-of-the-art FME algorithms, while its losses when compared with FSME, in terms of PSNR performance and computation time, are insignificant. We show that although our implementation platform is FPGA-based, our implementation results compare favourably with previous architectures implemented on ASICs. Finally we also show an improvement over some existing architectures implemented on FPGAs.

  12. 7 CFR 1951.264 - Action when borrower fails to cooperate, respond or graduate.

    Science.gov (United States)

    2010-01-01

    ... Analyzing Credit Needs and Graduation of Borrowers § 1951.264 Action when borrower fails to cooperate, respond or graduate. (a) When borrowers with other than FCP loans fail to: (1) Provide information... appeal the decision. (b) If an FCP borrower fails to cooperate after a lender expresses a willingness to...

  13. CYP264B1 from Sorangium cellulosum So ce56: a fascinating norisoprenoid and sesquiterpene hydroxylase.

    Science.gov (United States)

    Ly, Thuy T B; Khatri, Yogan; Zapp, Josef; Hutter, Michael C; Bernhardt, Rita

    2012-07-01

    Many terpenes and terpenoid compounds are known as bioactive substances with desirable fragrance and medicinal activities. Modification of such compounds to yield new derivatives with desired properties is particularly attractive. Cytochrome P450 monooxygenases are potential enzymes for these reactions due to their capability of performing different reactions on a variety of substrates. We report here the characterization of CYP264B1 from Sorangium cellulosum So ce56 as a novel sesquiterpene hydroxylase. CYP264B1 was able to convert various sesquiterpenes including nootkatone and norisoprenoids (α-ionone and β-ionone). Nootkatone, an important grapefruit aromatic sesquiterpenoid, was hydroxylated mainly at position C-13. The product has been shown to have the highest antiproliferative activity compared with other nootkatone derivatives. In addition, CYP264B1 was found to hydroxylate α- and β-ionone, important aroma compounds of floral scents, regioselectively at position C-3. The products, 3-hydroxy-β-ionone and 13-hydroxy-nootkatone, were confirmed by (1)H and (13)C NMR. The kinetics of the product formation was analyzed by high-performance liquid chromatography, and the K ( m ) and k (cat) values were calculated. The results of docking α-/β-ionone and nootkatone into a homology model of CYP264B1 revealed insights into the structural basis of these selective hydroxylations.

  14. Adaptive Rate Control Algorithm for H.264/AVC Considering Scene Change

    Directory of Open Access Journals (Sweden)

    Xiao Chen

    2013-01-01

    Full Text Available Scene change in H.264 video sequences has significant impact on the video communication quality. This paper presents a novel adaptive rate control algorithm with little additional calculation for H.264/AVC based on the scene change expression. According to the frame complexity quotiety, we define a scene change factor. It is used to allocate bits for each frame adaptively. Experimental results show that it can handle the scene change effectively. Our algorithm, in comparison to the JVT-G012 algorithm, reduces rate error and improves average peak signal-noise ratio with smaller deviation. It cannot only control bit rate accurately, but also get better video quality with the lower encoder buffer fullness to improve the quality of service.

  15. Edge-preserving Intra mode for efficient depth map coding based on H.264/AVC

    DEFF Research Database (Denmark)

    Zamarin, Marco; Forchhammer, Søren

    2014-01-01

    Depth-image-based-rendering (DIBR) algorithms for 3D video communication systems based on the “multi-view video plus depth” format are very sensitive to the accuracy of depth information. Specifically, edge regions in the depth data should be preserved in the coding/decoding process to ensure good...... view synthesis performance, which directly affects the overall system performance. This paper proposes a novel scheme for edge-aware Intra depth compression based on the H.264/AVC framework enabled on both Intra (I) and Inter (P) slices. The proposed scheme includes a new Intra mode specifically...... targeted to depth macroblocks with arbitrarily shaped edges, which are typically not predicted well by the standard Intra modes of H.264/AVC and result in high rate–distortion costs. The proposed algorithm segments edge macroblocks into two regions each approximated by a flat surface. A binary mask...

  16. Edge-preserving Intra Depth Coding based on Context-coding and H.264/AVC

    DEFF Research Database (Denmark)

    Zamarin, Marco; Salmistraro, Matteo; Forchhammer, Søren

    2013-01-01

    Depth map coding plays a crucial role in 3D Video communication systems based on the “Multi-view Video plus Depth” representation as view synthesis performance is strongly affected by the accuracy of depth information, especially at edges in the depth map image. In this paper an efficient algorithm...... for edge-preserving intra depth compression based on H.264/AVC is presented. The proposed method introduces a new Intra mode specifically targeted to depth macroblocks with arbitrarily shaped edges, which are typically not efficiently represented by DCT. Edge macroblocks are partitioned into two regions...... performance. Experiments show that the proposed Intra mode can improve view synthesis performance: average Bjøntegaard bit rate savings of 25% have been reported over a standard H.264/AVC Intra coder....

  17. A fast inter mode decision algorithm in H.264/AVC for IPTV broadcasting services

    Science.gov (United States)

    Kim, Geun-Yong; Yoon, Bin-Yeong; Ho, Yo-Sung

    2007-01-01

    The new video coding standard H.264/AVC employs the rate-distortion optimization (RDO) method for choosing the best coding mode. However, since it increases the encoder complexity tremendously, it is not suitable for real-time applications, such as IPTV broadcasting services. Therefore we need a fast mode decision algorithm to reduce its encoding time. In this paper, we propose a fast mode decision algorithm considering quantization parameter (QP) because we have noticed that the frequency of best modes depends on QP. In order to consider these characteristics, we use the coded block pattern (CBP) that has "0" value when all quantized discrete cosine transform (DCT) coefficients are zero. We also use both the early SKIP mode and early 16x16 mode decisions. Experimental results show that the proposed algorithm reduces the encoding time by 74.6% for the baseline profile and 72.8% for the main profile, compared to the H.264/AVC reference software.

  18. 4kUHD H264 Wireless Live Video Streaming Using CUDA

    Directory of Open Access Journals (Sweden)

    A. O. Adeyemi-Ejeye

    2014-01-01

    Full Text Available Ultrahigh definition video streaming has been explored in recent years. Most recently the possibility of 4kUHD video streaming over wireless 802.11n was presented, using preencoded video. Live encoding for streaming using x264 has proven to be very slow. The use of parallel encoding has been explored to speed up the process using CUDA. However there hasnot been a parallel implementation for video streaming. We therefore present for the first time a novel implementation of 4kUHD live encoding for streaming over a wireless network at low bitrate indoors, using CUDA for parallel H264 encoding. Our experimental results are used to verify our claim.

  19. USING H.264/AVC-INTRA FOR DCT BASED SEGMENTATION DRIVEN COMPOUND IMAGE COMPRESSION

    Directory of Open Access Journals (Sweden)

    S. Ebenezer Juliet

    2011-08-01

    Full Text Available This paper presents a one pass block classification algorithm for efficient coding of compound images which consists of multimedia elements like text, graphics and natural images. The objective is to minimize the loss of visual quality of text during compression by separating text information which needs high special resolution than the pictures and background. It segments computer screen images into text/graphics and picture/background classes based on DCT energy in each 4x4 block, and then compresses both text/graphics pixels and picture/background blocks by H.264/AVC with variable quantization parameter. Experimental results show that the single H.264/AVC-INTRA coder with variable quantization outperforms single coders such as JPEG, JPEG-2000 for compound images. Also the proposed method improves the PSNR value significantly than standard JPEG, JPEG-2000 and while keeping competitive compression ratios.

  20. Low-complexity transcoding algorithm from H.264/AVC to SVC using data mining

    Science.gov (United States)

    Garrido-Cantos, Rosario; De Cock, Jan; Martínez, Jose Luis; Van Leuven, Sebastian; Cuenca, Pedro; Garrido, Antonio

    2013-12-01

    Nowadays, networks and terminals with diverse characteristics of bandwidth and capabilities coexist. To ensure a good quality of experience, this diverse environment demands adaptability of the video stream. In general, video contents are compressed to save storage capacity and to reduce the bandwidth required for its transmission. Therefore, if these compressed video streams were compressed using scalable video coding schemes, they would be able to adapt to those heterogeneous networks and a wide range of terminals. Since the majority of the multimedia contents are compressed using H.264/AVC, they cannot benefit from that scalability. This paper proposes a low-complexity algorithm to convert an H.264/AVC bitstream without scalability to scalable bitstreams with temporal scalability in baseline and main profiles by accelerating the mode decision task of the scalable video coding encoding stage using machine learning tools. The results show that when our technique is applied, the complexity is reduced by 87% while maintaining coding efficiency.

  1. Continuous electrochemical monitoring of nitric oxide production in murine macrophage cell line RAW 264.7

    Czech Academy of Sciences Publication Activity Database

    Pekarová, Michaela; Králová, Jana; Kubala, Lukáš; Číž, Milan; Lojek, Antonín; Gregor, Č.; Hrbáč, J.

    2009-01-01

    Roč. 394, č. 5 (2009), s. 1497-1504 ISSN 1618-2642 R&D Projects: GA AV ČR(CZ) 1QS500040507 Grant - others:GA ČR(CZ) GP524/05/P135 Institutional research plan: CEZ:AV0Z50040507; CEZ:AV0Z50040702 Keywords : nitric oxide * macrophage s RAW 264.7 * nitric oxide sensor Subject RIV: BO - Biophysics Impact factor: 3.480, year: 2009

  2. Autophagy in RAW264.7 Cells Treated with Surface-Functionalized Graphene Oxides

    Directory of Open Access Journals (Sweden)

    Chang Seok Park

    2015-01-01

    Full Text Available This study investigated cytotoxicity, particularly autophagy, in RAW264.7 cells exposed to graphene oxide (GO and its derivatives (dodecylamine-GO (DA-GO, reduced GO (rGO, and sodium dodecyl sulfate-rGO (SDS-rGO. Appearance of amine stretching bands, out-of-plane C-H stretching vibrations, and S=O stretching bands in infrared spectra indicated the formation of DA-GO, rGO, and SDS-rGO, respectively. Light microscopy and microculture tetrazolium assay showed that all the GO types exerted cytotoxic effects on RAW264.7 cells in a concentration-dependent manner. Higher concentrations of the GO types downregulated the expression of PU.1, a unique transcription factor in monocytes and macrophages, and decreased the conversion of LC3A/B-I to LC3A/B-II, suggesting that PU.1 was associated with autophagy in RAW264.7 cells. These results suggested that surface-functionalized GOs exerted cytotoxic effects in a concentration-dependent manner by changing the expression of critical genes and inducing autophagy in macrophages.

  3. Streptococcus pyogenes CAMP factor attenuates phagocytic activity of RAW 264.7 cells.

    Science.gov (United States)

    Kurosawa, Mie; Oda, Masataka; Domon, Hisanori; Saitoh, Issei; Hayasaki, Haruaki; Terao, Yutaka

    2016-02-01

    Streptococcus pyogenes produces molecules that inhibit the function of human immune system, thus allowing the pathogen to grow and spread in tissues. It is known that S. pyogenes CAMP factor increases erythrocytosis induced by Staphylococcus aureus β-hemolysin. However, the effects of CAMP factor for immune cells are unclear. In this study, we investigated the effects of CAMP factor to macrophages. Western blotting analysis demonstrated that all examined strains expressed CAMP factor protein. In the presence of calcium or magnesium ion, CAMP factor was significantly released in the supernatant. In addition, both culture supernatant from S. pyogenes strain SSI-9 and recombinant CAMP factor dose-dependently induced vacuolation in RAW 264.7 cells, but the culture supernatant from Δcfa isogenic mutant strain did not. CAMP factor formed oligomers in RAW 264.7 cells in a time-dependent manner. CAMP factor suppressed cell proliferation via G2 phase cell cycle arrest without inducing cell death. Furthermore, CAMP factor reduced the uptake of S. pyogenes and phagocytic activity indicator by RAW 264.7 cells. These results suggest that CAMP factor works as a macrophage dysfunction factor. Therefore, we conclude that CAMP factor allows S. pyogenes to escape the host immune system, and contribute to the spread of streptococcal infection. Copyright © 2015 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.

  4. Simvastatin induces caspase-independent apoptosis in LPS-activated RAW264.7 macrophage cells

    International Nuclear Information System (INIS)

    Kim, Yong Chan; Song, Seok Bean; Lee, Mi Hee; Kang, Kwang Il; Lee, Hayyoung; Paik, Sang-Gi; Kim, Kyoon Eon; Kim, Young Sang

    2006-01-01

    Macrophages participate in several inflammatory pathologies such as sepsis and arthritis. We examined the effect of simvastatin on the LPS-induced proinflammatory macrophage RAW264.7 cells. Co-treatment of LPS and a non-toxic dose of simvastatin induced cell death in RAW264.7 cells. The cell death was accompanied by disruption of mitochondrial membrane potential (MMP), genomic DNA fragmentation, and caspase-3 activation. Surprisingly, despite caspase-dependent apoptotic cascade being completely blocked by Z-VAD-fmk, a pan-caspase inhibitor, the cell death was only partially repressed. In the presence of Z-VAD-fmk, DNA fragmentation was blocked, but DNA condensation, disruption of MMP, and nuclear translocation of apoptosis inducing factor were obvious. The cell death by simvastatin and LPS was effectively decreased by both the FPP and GGPP treatments as well as mevalonate. Our findings indicate that simvastatin triggers the cell death of LPS-treated RAW264.7 cells through both caspase-dependent and -independent apoptotic pathways, suggesting a novel mechanism of statins for the severe inflammatory disease therapy

  5. IM-133N modulates cytokine secretion by RAW264.7 and THP-1 cells.

    Science.gov (United States)

    Varma, R Sandeep; Guruprasad, Kanive P; Satyamoorthy, Kapaettu; Kumar, L M Sharath; Babu, U Venkanna; Patki, S Pralhad

    2016-01-01

    An indigenous herbal extract IM-133N containing extracts of Prosopis glandulosa Torr and Symplocos racemosa Roxb were evaluated for potential immunomodulatory effects using RAW264.7 and THP-1 cells. The incubation of the cells for 24 h with IM-133N over a dose range 0-125 µg/ml did not cause cytotoxicity that exceeded 10%. The results indicated that non-cytotoxic doses of IM-133N effectively up-regulated iNOS, TNFα, IL-6, IL-10, IL-8 and IFNγ gene expression in both the RAW264.7 and THP-1 cells. The results also indicated IM-133N elicited dose-related increases in nitric oxide (NO) and tumor necrosis factor (TNF)-α production by RAW264.7 or THP-1 cells. These results demonstrated that IM-133N could stimulate NO and induced pro-inflammatory cytokine expression by monocytes/macrophages. As clinical studies have shown IM-133N to be an effective immunomodulator without any adverse effects, the results of the present study provide further support for the potential use of this agent as an immunostimulant or as an immunotherapy adjuvant.

  6. Efficient Transmission of H.264 Video over Multirate IEEE 802.11e WLANs

    Directory of Open Access Journals (Sweden)

    Alnuweiri Hussein

    2008-01-01

    Full Text Available The H.264 video encoding technology, which has emerged as one of the most promising compression standards, offers many new delivery-aware features such as data partitioning. Efficient transmission of H.264 video over any communication medium requires a great deal of coordination between different communication network layers. This paper considers the increasingly popular and widespread 802.11 Wireless Local Area Networks (WLANs and studies different schemes for the delivery of the baseline and extended profiles of H.264 video over such networks. While the baseline profile produces data similar to conventional video technologies, the extended profile offers a partitioning feature that divides video data into three sets with different levels of importance. This allows for the use of service differentiation provided in the WLAN. This paper examines the video transmission performance of the existing contention-based solutions for 802.11e, and compares it to our proposed scheduled access mechanism. It is demonstrated that the scheduled access scheme outperforms contention-based prioritized services of the 802.11e standard. For partitioned video, it is shown that the overhead of partitioning is too high, and better results are achieved if some partitions are aggregated. The effect of link adaptation and multirate operation of the physical layer (PHY is also investigated in this paper.

  7. Compress compound images in H.264/MPGE-4 AVC by exploiting spatial correlation.

    Science.gov (United States)

    Lan, Cuiling; Shi, Guangming; Wu, Feng

    2010-04-01

    Compound images are a combination of text, graphics and natural image. They present strong anisotropic features, especially on the text and graphics parts. These anisotropic features often render conventional compression inefficient. Thus, this paper proposes a novel coding scheme from the H.264 intraframe coding. In the scheme, two new intramodes are developed to better exploit spatial correlation in compound images. The first is the residual scalar quantization (RSQ) mode, where intrapredicted residues are directly quantized and coded without transform. The second is the base colors and index map (BCIM) mode that can be viewed as an adaptive color quantization. In this mode, an image block is represented by several representative colors, referred to as base colors, and an index map to compress. Every block selects its coding mode from two new modes and the previous intramodes in H.264 by rate-distortion optimization (RDO). Experimental results show that the proposed scheme improves the coding efficiency even more than 10 dB at most bit rates for compound images and keeps a comparable efficient performance to H.264 for natural images.

  8. Efficient MPEG-2 to H.264/AVC Transcoding of Intra-Coded Video

    Directory of Open Access Journals (Sweden)

    Vetro Anthony

    2007-01-01

    Full Text Available This paper presents an efficient transform-domain architecture and corresponding mode decision algorithms for transcoding intra-coded video from MPEG-2 to H.264/AVC. Low complexity is achieved in several ways. First, our architecture employs direct conversion of the transform coefficients, which eliminates the need for the inverse discrete cosine transform (DCT and forward H.264/AVC transform. Then, within this transform-domain architecture, we perform macroblock-based mode decisions based on H.264/AVC transform coefficients, which is possible using a novel method of calculating distortion in the transform domain. The proposed method for distortion calculation could be used to make rate-distortion optimized mode decisions with lower complexity. Compared to the pixel-domain architecture with rate-distortion optimized mode decision, simulation results show that there is a negligible loss in quality incurred by the direct conversion of transform coefficients and the proposed transform-domain mode decision algorithms, while complexity is significantly reduced. To further reduce the complexity, we also propose two fast mode decision algorithms. The first algorithm ranks modes based on a simple cost function in the transform domain, then computes the rate-distortion optimal mode from a reduced set of ranked modes. The second algorithm exploits temporal correlations in the mode decision between temporally adjacent frames. Simulation results show that these algorithms provide additional computational savings over the proposed transform-domain architecture while maintaining virtually the same coding efficiency.

  9. Anti-inflammatory Potential of Petiveria alliacea on Activated RAW264.7 Murine Macrophages.

    Science.gov (United States)

    Gutierrez, Rosa Martha Perez; Hoyo-Vadillo, Carlos

    2017-07-01

    Defense and protection to multiple harmful stimuli are the inflammation, when is self-amplified and uncontrolled is the basis of the pathogenesis of a wide variety of inflammatory illness. The aim of this study was to evaluate if Petiveria alliacea could attenuate inflammation in a murine model of RAW264 macrophages the involved model and its involved mechanism. The ethanol extract from P. alliacea was precipitated with water and supernatant was used for this study (PW). The anti-inflammatory effects of PW were investigated through evaluating of the production of several cytokines, chemokines, and expression of nuclear factor-kappa B (NF-κB) in lipopolysaccharide (LPS)-stimulated RAW264.7 macrophages. Also was determined the ability to decrease the oxidative stress in RAW264.7 cells with carboxy-2',7'-dichloro-dihydro-fluorescein diacetate. PW significantly suppress the secretion of prostaglandin E 2 , leukotriene C 4 , interleukin (IL)-1 β, IL-6, IL-10, interferon gamma nitric oxide (NO), inducible NO synthase, IL-1 β, IL-4, in RAW264.7 cells in a dose-dependent manner. In addition, PW also markedly inhibited the transcriptional activity of NF-κB. PW produced significant anti-inflammatory activity through inhibiting the production of inflammatory mediators through the NF-κB inactivation in the LPS-stimulated RAW24.7 cells. PW exerts significant antioxidant and anti-inflammatory activities, and this effect can be attributed in part, to the presence of dibenzyl disulfide, dibenzyl trisulfide pinitol, coumarin, myricetin, glutamyl-S-benzyl cysteine, and petiveriins A and B. Treatment with ethanol extract from Petiveria alliacea which was previously precipitated with water and supernatant (PE) was tested in LPS-stimulated RAW264.7 cells. PE suppressed the level of oxidative stress and the induction of proinflammatory mediators, as PGE2, LTC4, IL-1 ß, IL-6, IL-10, IFN- NO, iNOS, IL-1 ß, IL-4, in RAW264.7 macrophages through NF-B inactivation. These findings

  10. Application Layer Systematic Network Coding for Sliced H.264/AVC Video Streaming

    Directory of Open Access Journals (Sweden)

    Sajid Nazir

    2012-01-01

    Full Text Available Application Layer Forward Error Correction (AL-FEC with rateless codes can be applied to protect the video data over lossy channels. Expanding Window Random Linear Codes (EW RLCs are a flexible unequal error protection fountain coding scheme which can provide prioritized data transmission. In this paper, we propose a system that exploits systematic EW RLC for H.264/Advanced Video Coding (AVC slice-partitioned data. The system prioritizes slices based on their PSNR contribution to reconstruction as well as temporal significance. Simulation results demonstrate usefulness of using relative slice priority with systematic codes for multimedia broadcast applications.

  11. Efficient depth intraprediction method for H.264/AVC-based three-dimensional video coding

    Science.gov (United States)

    Oh, Kwan-Jung; Oh, Byung Tae

    2015-04-01

    We present an intracoding method that is applicable to depth map coding in multiview plus depth systems. Our approach combines skip prediction and plane segmentation-based prediction. The proposed depth intraskip prediction uses the estimated direction at both the encoder and decoder, and does not need to encode residual data. Our plane segmentation-based intraprediction divides the current block into biregions, and applies a different prediction scheme for each segmented region. This method avoids incorrect estimations across different regions, resulting in higher prediction accuracy. Simulation results demonstrate that the proposed scheme is superior to H.264/advanced video coding intraprediction and has the ability to improve the subjective rendering quality.

  12. Subjective Video Quality Assessment in H.264/AVC Video Coding Standard

    Directory of Open Access Journals (Sweden)

    Z. Miličević

    2012-11-01

    Full Text Available This paper seeks to provide an approach for subjective video quality assessment in the H.264/AVC standard. For this purpose a special software program for the subjective assessment of quality of all the tested video sequences is developed. It was developed in accordance with recommendation ITU-T P.910, since it is suitable for the testing of multimedia applications. The obtained results show that in the proposed selective intra prediction and optimized inter prediction algorithm there is a small difference in picture quality (signal-to-noise ratio between decoded original and modified video sequences.

  13. Anti-Inflammatory Effect of Lupinalbin A Isolated from Apios americana on Lipopolysaccharide-Treated RAW264.7 Cells

    Directory of Open Access Journals (Sweden)

    Hyo-Young Kim

    2018-03-01

    Full Text Available Apios americana, a leguminous plant, is used as food in some countries. Although the biological activities of Apios extract have been reported, there have been no reports about the anti-inflammatory mechanism of lupinalbin A on the RAW264.7 cells. In this study, we investigated the anti-inflammatory effect of A. americana lupinalbin A on lipopolysaccharide (LPS-treated RAW264.7 cells. Lupinalbin A significantly inhibited nitric oxide production and inducible nitric oxide synthase expression in LPS-treated RAW264.7 cells. The expression of cytokines, including interleukin-6, tumor necrosis factor-α, and chemokine of monocyte chemoattractant protein, was reduced under lupinalbin A exposure in LPS-treated RAW264.7 cells. In addition, lupinalbin A significantly decreased LPS-induced interferon (IFN-β production and STAT1 protein levels in RAW264.7 cells. Taken together, these results suggest that A. americana lupinalbin A exerts anti-inflammatory effects via the inhibition of pro-inflammatory cytokines and blocking of IFN-β/STAT1 pathway activation.

  14. Video Traffic Characteristics of Modern Encoding Standards: H.264/AVC with SVC and MVC Extensions and H.265/HEVC

    Directory of Open Access Journals (Sweden)

    Patrick Seeling

    2014-01-01

    Full Text Available Video encoding for multimedia services over communication networks has significantly advanced in recent years with the development of the highly efficient and flexible H.264/AVC video coding standard and its SVC extension. The emerging H.265/HEVC video coding standard as well as 3D video coding further advance video coding for multimedia communications. This paper first gives an overview of these new video coding standards and then examines their implications for multimedia communications by studying the traffic characteristics of long videos encoded with the new coding standards. We review video coding advances from MPEG-2 and MPEG-4 Part 2 to H.264/AVC and its SVC and MVC extensions as well as H.265/HEVC. For single-layer (nonscalable video, we compare H.265/HEVC and H.264/AVC in terms of video traffic and statistical multiplexing characteristics. Our study is the first to examine the H.265/HEVC traffic variability for long videos. We also illustrate the video traffic characteristics and statistical multiplexing of scalable video encoded with the SVC extension of H.264/AVC as well as 3D video encoded with the MVC extension of H.264/AVC.

  15. Video traffic characteristics of modern encoding standards: H.264/AVC with SVC and MVC extensions and H.265/HEVC.

    Science.gov (United States)

    Seeling, Patrick; Reisslein, Martin

    2014-01-01

    Video encoding for multimedia services over communication networks has significantly advanced in recent years with the development of the highly efficient and flexible H.264/AVC video coding standard and its SVC extension. The emerging H.265/HEVC video coding standard as well as 3D video coding further advance video coding for multimedia communications. This paper first gives an overview of these new video coding standards and then examines their implications for multimedia communications by studying the traffic characteristics of long videos encoded with the new coding standards. We review video coding advances from MPEG-2 and MPEG-4 Part 2 to H.264/AVC and its SVC and MVC extensions as well as H.265/HEVC. For single-layer (nonscalable) video, we compare H.265/HEVC and H.264/AVC in terms of video traffic and statistical multiplexing characteristics. Our study is the first to examine the H.265/HEVC traffic variability for long videos. We also illustrate the video traffic characteristics and statistical multiplexing of scalable video encoded with the SVC extension of H.264/AVC as well as 3D video encoded with the MVC extension of H.264/AVC.

  16. Low-cost VLSI architecture design for forward quantization of H.264/AVC

    Science.gov (United States)

    Ruiz, G. A.; Michell, J. A.

    2007-05-01

    The H.264/AVC (Advanced Video Codec) is the latest standard for video coding. It assumes a scalar forward quantizer performed at the encoder which can be implemented directly in integer arithmetic. An efficient architecture for the computation of forward quantization of H.264/AVC is presented in this paper. It uses a modification of the quantization operation which reduces the arithmetic operations, and a truncated Booth multiplier based on adaptative statistical approach, which reduces the hardware. The JM reference software's C code has been re-written to analyze the effect of new algorithm and of truncated Booth multiplier. Simulations made up over popular test sequences used in video standardization show the validity of this approach. These results demonstrate that, at low QP, the PSNR is improved between a maximum of +0.81db and a minimum of 0.31db, with a slight increase in the Bit Rate being around 0.8%. Finally, a suitable architecture for VLSI implementation is presented, which reduces in a 26% the area, 32% the power and 21% the critical path delay in comparison with classical implementation. Moreover, it also reduces the area and increase the speed in comparison with architectures presented in references.

  17. Low-Power Bitstream-Residual Decoder for H.264/AVC Baseline Profile Decoding

    Directory of Open Access Journals (Sweden)

    Xu Ke

    2009-01-01

    Full Text Available Abstract We present the design and VLSI implementation of a novel low-power bitstream-residual decoder for H.264/AVC baseline profile. It comprises a syntax parser, a parameter decoder, and an Inverse Quantization Inverse Transform (IQIT decoder. The syntax parser detects and decodes each incoming codeword in the bitstream under the control of a hierarchical Finite State Machine (FSM; the IQIT decoder performs inverse transform and quantization with pipelining and parallelism. Various power reduction techniques, such as data-driven based on statistic results, nonuniform partition, precomputation, guarded evaluation, hierarchical FSM decomposition, TAG method, zero-block skipping, and clock gating , are adopted and integrated throughout the bitstream-residual decoder. With innovative architecture, the proposed design is able to decode QCIF video sequences of 30 fps at a clock rate as low as 1.5 MHz. A prototype H.264/AVC baseline decoding chip utilizing the proposed decoder is fabricated in UMC 0.18  m 1P6M CMOS technology. The proposed design is measured under 1 V 1.8 V supply with 0.1 V step. It dissipates 76  W at 1 V and 253  W at 1.8 V.

  18. Immunomodulatory effects of an acetylated Cyclocarya paliurus polysaccharide on murine macrophages RAW264.7.

    Science.gov (United States)

    Liu, Xin; Xie, Jianhua; Jia, Shuo; Huang, Lixin; Wang, Zhijun; Li, Chang; Xie, Mingyong

    2017-05-01

    Polysaccharides (CP) extracted from the leaves of Cyclocarya paliurus (C. paliurus) have been shown to possess a variety of biological activities. In present study, CP was successfully modified to obtain its acetylated derivative Ac-CP. Its potential immunomodulatory activities on RAW264.7 macrophages were investigated. Results showed that the acetylated polysaccharide Ac-CP could significantly stimulate macrophage proliferation, its actions were significantly stronger than that of the corresponding unmodified polysaccharide, CP. Meanwhile, the NO production activities of macrophages were not significantly enhanced by Ac-CP compared to CP group. In addition, both the phagocytic activity and levels of cytokines TNF-a, IL-1β and IL-6 were enhanced in the RAW264.7 macrophages by stimulation of Ac-CP. These results indicated that the acetylated derivative Ac-CP could enhance the activation of peritoneal macrophages, and acetylation modification can enhance the immunomodulation function of CP, indicating the potential application of acetylated polysaccharide as an immunotherapeutic adjuvant. Copyright © 2017 Elsevier B.V. All rights reserved.

  19. Neuropeptide FF inhibits LPS-mediated osteoclast differentiation of RAW264.7 cells.

    Science.gov (United States)

    Sun, Yu-Long; Chen, Zhi-Hao; Li, Di-Jie; Zhao, Fan; Ma, Xiao-Li; Shang, Peng; Yang, Tuanming; Qian, Airong

    2014-01-01

    Neuropeptide FF (NPFF) has been implicated in many physiological processes. Previously, we have reported that NPFF modulates the viability and nitric oxide (NO) production of RAW264.7 macrophages. In this study, we investigated the influence of NPFF on lipopolysaccharide (LPS)-mediated osteoclast formation of RAW264.7 cells. Our results suggest that, NPFF dose-dependently (1 nM, 10 nM and 100 nM) inhibited osteoclast formation, TRAP enzyme activity and bone resorption in osteoclasts induced by LPS respectively. Moreover, LPS-provoked NO release was also inhibited by NPFF treatment, indicating a NO-dependent pathway is mainly involved. Furthermore, the alterations of osteoclast marker genes were also assessed including TRAP, Cathepsin K, MMP-9, NFATc1 and Runx2. NPFF downregulated LPS-caused gene augmentations of TRAP, Cathepsin K and MMP-9, whereas showed no influences on NFATc1 and Runx2. In addition, NPFF receptor 2 (NPFFR2) mRNA expression was also augmented in response to NPFF treatment, hinting the involvement of NPFFR2 pathway. It should be mentioned that RF9 (1 µ M), a reported pharmacological inhibitor for NPFF receptors, exerted NPFF-like agonist properties as to attenuate osteoclastogenesis. Collectively, our findings provide new evidence for the in vitro activity of NPFF on osteoclasts, which may be helpful to extend the scope of NPFF functions.

  20. A Novel Steganography Technique for SDTV-H.264/AVC Encoded Video

    Directory of Open Access Journals (Sweden)

    Christian Di Laura

    2016-01-01

    Full Text Available Today, eavesdropping is becoming a common issue in the rapidly growing digital network and has foreseen the need for secret communication channels embedded in digital media. In this paper, a novel steganography technique designed for Standard Definition Digital Television (SDTV H.264/AVC encoded video sequences is presented. The algorithm introduced here makes use of the compression properties of the Context Adaptive Variable Length Coding (CAVLC entropy encoder to achieve a low complexity and real-time inserting method. The chosen scheme hides the private message directly in the H.264/AVC bit stream by modifying the AC frequency quantized residual luminance coefficients of intrapredicted I-frames. In order to avoid error propagation in adjacent blocks, an interlaced embedding strategy is applied. Likewise, the steganography technique proposed allows self-detection of the hidden message at the target destination. The code source was implemented by mixing MATLAB 2010 b and Java development environments. Finally, experimental results have been assessed through objective and subjective quality measures and reveal that less visible artifacts are produced with the technique proposed by reaching PSNR values above 40.0 dB and an embedding bit rate average per secret communication channel of 425 bits/sec. This exemplifies that steganography is affordable in digital television.

  1. Evaluation of H.264/AVC over IEEE 802.11p vehicular networks

    Science.gov (United States)

    Rozas-Ramallal, Ismael; Fernández-Caramés, Tiago M.; Dapena, Adriana; García-Naya, José Antonio

    2013-12-01

    The capacity of vehicular networks to offer non-safety services, like infotainment applications or the exchange of multimedia information between vehicles, have attracted a great deal of attention to the field of Intelligent Transport Systems (ITS). In particular, in this article we focus our attention on IEEE 802.11p which defines enhancements to IEEE 802.11 required to support ITS applications. We present an FPGA-based testbed developed to evaluate H.264/AVC (Advanced Video Coding) video transmission over vehicular networks. The testbed covers some of the most common situations in vehicle-to-vehicle and roadside-to-vehicle communications and it is highly flexible, allowing the performance evaluation of different vehicular standard configurations. We also show several experimental results to illustrate the quality obtained when H.264/AVC encoded video is transmitted over IEEE 802.11p networks. The quality is measured considering two important parameters: the percentage of recovered group of pictures and the frame quality. In order to improve performance, we propose to substitute the convolutional channel encoder used in IEEE 802.11p for a low-density parity-check code encoder. In addition, we suggest a simple strategy to decide the optimum number of iterations needed to decode each packet received.

  2. Context adaptive binary arithmetic coding-based data hiding in partially encrypted H.264/AVC videos

    Science.gov (United States)

    Xu, Dawen; Wang, Rangding

    2015-05-01

    A scheme of data hiding directly in a partially encrypted version of H.264/AVC videos is proposed which includes three parts, i.e., selective encryption, data embedding and data extraction. Selective encryption is performed on context adaptive binary arithmetic coding (CABAC) bin-strings via stream ciphers. By careful selection of CABAC entropy coder syntax elements for selective encryption, the encrypted bitstream is format-compliant and has exactly the same bit rate. Then a data-hider embeds the additional data into partially encrypted H.264/AVC videos using a CABAC bin-string substitution technique without accessing the plaintext of the video content. Since bin-string substitution is carried out on those residual coefficients with approximately the same magnitude, the quality of the decrypted video is satisfactory. Video file size is strictly preserved even after data embedding. In order to adapt to different application scenarios, data extraction can be done either in the encrypted domain or in the decrypted domain. Experimental results have demonstrated the feasibility and efficiency of the proposed scheme.

  3. Subjective Quality Assessment of H.264/AVC Video Streaming with Packet Losses

    Directory of Open Access Journals (Sweden)

    Naccari Matteo

    2011-01-01

    Full Text Available Research in the field of video quality assessment relies on the availability of subjective scores, collected by means of experiments in which groups of people are asked to rate the quality of video sequences. The availability of subjective scores is fundamental to enable validation and comparative benchmarking of the objective algorithms that try to predict human perception of video quality by automatically analyzing the video sequences, in a way to support reproducible and reliable research results. In this paper, a publicly available database of subjective quality scores and corrupted video sequences is described. The scores refer to 156 sequences at CIF and 4CIF spatial resolutions, encoded with H.264/AVC and corrupted by simulating the transmission over an error-prone network. The subjective evaluation has been performed by 40 subjects at the premises of two academic institutions, in standard-compliant controlled environments. In order to support reproducible research in the field of full-reference, reduced-reference, and no-reference video quality assessment algorithms, both the uncompressed files and the H.264/AVC bitstreams, as well as the packet loss patterns, have been made available to the research community.

  4. Video Quality Prediction Models Based on Video Content Dynamics for H.264 Video over UMTS Networks

    Directory of Open Access Journals (Sweden)

    Asiya Khan

    2010-01-01

    Full Text Available The aim of this paper is to present video quality prediction models for objective non-intrusive, prediction of H.264 encoded video for all content types combining parameters both in the physical and application layer over Universal Mobile Telecommunication Systems (UMTS networks. In order to characterize the Quality of Service (QoS level, a learning model based on Adaptive Neural Fuzzy Inference System (ANFIS and a second model based on non-linear regression analysis is proposed to predict the video quality in terms of the Mean Opinion Score (MOS. The objective of the paper is two-fold. First, to find the impact of QoS parameters on end-to-end video quality for H.264 encoded video. Second, to develop learning models based on ANFIS and non-linear regression analysis to predict video quality over UMTS networks by considering the impact of radio link loss models. The loss models considered are 2-state Markov models. Both the models are trained with a combination of physical and application layer parameters and validated with unseen dataset. Preliminary results show that good prediction accuracy was obtained from both the models. The work should help in the development of a reference-free video prediction model and QoS control methods for video over UMTS networks.

  5. DISTRIBUTION OF CCR2-64I GENE AMONG THE TRIBES AND CASTE POPULATION OF VIDARBHA, INDIA.

    Directory of Open Access Journals (Sweden)

    Arvind B Chavhan

    2013-08-01

    Results: The genotyping for the CCR2-64I mutation among the selected tribe and a caste reveal that all of the tribes and a caste was found to be heterozygous for the CCR2-64I mutation. Among the tribes Gonds showed highest genotype frequency (29.28% and (11.76% for heterozygous (CCR2/64I and Homozygous (64I/64I respectively, having an allelic frequency (0.233. A pooled allelic frequencies of the wild-type allele CCR2 and CCR2 64I the variant were found to be 0.854 and 0.146, respectively. No significant deviations from the HWE were observed for tribes and a caste population for the CCR2- 64I mutant χ2=2.76. The study reports the presence of mutant CCR2- 64I gene in tribes and caste population from Vidarbha region.

  6. Encoder power consumption comparison of Distributed Video Codec and H.264/AVC in low-complexity mode

    DEFF Research Database (Denmark)

    Ukhanova, Ann; Belyaev, Evgeny; Forchhammer, Søren

    2010-01-01

    This paper presents a power consumption comparison of a novel approach to video compression based on distributed video coding (DVC) and widely used video compression based on H.264/AVC standard. We have used a low-complexity configuration for H.264/AVC codec. It is well-known that motion estimation...... stated in this paper show that for current implementations of DVC these statements could be disputed from a power consumption/compression efficiency point of view when comparing to compression algorithms based on differential frame coding (with zero search radius for ME)....

  7. Parallel processing architecture for H.264 deblocking filter on multi-core platforms

    Science.gov (United States)

    Prasad, Durga P.; Sonachalam, Sekar; Kunchamwar, Mangesh K.; Gunupudi, Nageswara Rao

    2012-03-01

    Massively parallel computing (multi-core) chips offer outstanding new solutions that satisfy the increasing demand for high resolution and high quality video compression technologies such as H.264. Such solutions not only provide exceptional quality but also efficiency, low power, and low latency, previously unattainable in software based designs. While custom hardware and Application Specific Integrated Circuit (ASIC) technologies may achieve lowlatency, low power, and real-time performance in some consumer devices, many applications require a flexible and scalable software-defined solution. The deblocking filter in H.264 encoder/decoder poses difficult implementation challenges because of heavy data dependencies and the conditional nature of the computations. Deblocking filter implementations tend to be fixed and difficult to reconfigure for different needs. The ability to scale up for higher quality requirements such as 10-bit pixel depth or a 4:2:2 chroma format often reduces the throughput of a parallel architecture designed for lower feature set. A scalable architecture for deblocking filtering, created with a massively parallel processor based solution, means that the same encoder or decoder will be deployed in a variety of applications, at different video resolutions, for different power requirements, and at higher bit-depths and better color sub sampling patterns like YUV, 4:2:2, or 4:4:4 formats. Low power, software-defined encoders/decoders may be implemented using a massively parallel processor array, like that found in HyperX technology, with 100 or more cores and distributed memory. The large number of processor elements allows the silicon device to operate more efficiently than conventional DSP or CPU technology. This software programing model for massively parallel processors offers a flexible implementation and a power efficiency close to that of ASIC solutions. This work describes a scalable parallel architecture for an H.264 compliant deblocking

  8. 20 CFR 1002.264 - Is the employee allowed to repay a previous distribution from a pension benefits plan upon being...

    Science.gov (United States)

    2010-04-01

    ... 20 Employees' Benefits 3 2010-04-01 2010-04-01 false Is the employee allowed to repay a previous distribution from a pension benefits plan upon being reemployed? 1002.264 Section 1002.264 Employees' Benefits OFFICE OF THE ASSISTANT SECRETARY FOR VETERANS' EMPLOYMENT AND TRAINING SERVICE, DEPARTMENT OF LABOR REGULATIONS UNDER THE UNIFORMED...

  9. A perceptual-based approach to bit allocation for H.264 encoder

    Science.gov (United States)

    Ou, Tao-Sheng; Huang, Yi-Hsin; Chen, Homer H.

    2010-07-01

    Since the ultimate receivers of encoded video are human eyes, the characteristics of human visual system should be taken into consideration in the design of bit allocation to improve the perceptual video quality. In this paper, we incorporate the structural similarity index as a distortion metric and propose a novel rate-distortion model to characterize the relationship between rate and the structural similarity index. Based on the model, we develop an optimum bit allocation and rate control scheme for H.264 encoders. Experimental results show that up to 25% bitrate reduction over the JM reference software can be achieved. Subjective evaluation further confirms that the proposed scheme preserves more structural information and improves the perceptual quality of the encoded video.

  10. Lipopolysaccharide from Burkholderia thailandensis E264 provides protection in a murine model of melioidosis.

    Science.gov (United States)

    Ngugi, Sarah A; Ventura, Valeria V; Qazi, Omar; Harding, Sarah V; Kitto, G Barrie; Estes, D Mark; Dell, Anne; Titball, Richard W; Atkins, Timothy P; Brown, Katherine A; Hitchen, Paul G; Prior, Joann L

    2010-11-03

    Burkholderia thailandensis is a less virulent close relative of Burkholderia pseudomallei, a CDC category B biothreat agent. We have previously shown that lipopolysaccharide (LPS) extracted from B. pseudomallei can provide protection against a lethal challenge of B. pseudomallei in a mouse model of melioidosis. Sugar analysis on LPS from B. thailandensis strain E264 confirmed that this polysaccharide has a similar structure to LPS from B. pseudomallei. Mice were immunised with LPS from B. thailandensis or B. pseudomallei and challenged with a lethal dose of B. pseudomallei strain K96243. Similar protection levels were observed when either LPS was used as the immunogen. This data suggests that B. thailandensis LPS has the potential to be used as part of a subunit based vaccine against pathogenic B. pseudomallei. Crown Copyright © 2010. Published by Elsevier Ltd. All rights reserved.

  11. A Robust H.264/AVC Video Watermarking Scheme with Drift Compensation

    Directory of Open Access Journals (Sweden)

    Xinghao Jiang

    2014-01-01

    Full Text Available A robust H.264/AVC video watermarking scheme for copyright protection with self-adaptive drift compensation is proposed. In our scheme, motion vector residuals of macroblocks with the smallest partition size are selected to hide copyright information in order to hold visual impact and distortion drift to a minimum. Drift compensation is also implemented to reduce the influence of watermark to the most extent. Besides, discrete cosine transform (DCT with energy compact property is applied to the motion vector residual group, which can ensure robustness against intentional attacks. According to the experimental results, this scheme gains excellent imperceptibility and low bit-rate increase. Malicious attacks with different quantization parameters (QPs or motion estimation algorithms can be resisted efficiently, with 80% accuracy on average after lossy compression.

  12. Game theoretic wireless resource allocation for H.264 MGS video transmission over cognitive radio networks

    Science.gov (United States)

    Fragkoulis, Alexandros; Kondi, Lisimachos P.; Parsopoulos, Konstantinos E.

    2015-03-01

    We propose a method for the fair and efficient allocation of wireless resources over a cognitive radio system network to transmit multiple scalable video streams to multiple users. The method exploits the dynamic architecture of the Scalable Video Coding extension of the H.264 standard, along with the diversity that OFDMA networks provide. We use a game-theoretic Nash Bargaining Solution (NBS) framework to ensure that each user receives the minimum video quality requirements, while maintaining fairness over the cognitive radio system. An optimization problem is formulated, where the objective is the maximization of the Nash product while minimizing the waste of resources. The problem is solved by using a Swarm Intelligence optimizer, namely Particle Swarm Optimization. Due to the high dimensionality of the problem, we also introduce a dimension-reduction technique. Our experimental results demonstrate the fairness imposed by the employed NBS framework.

  13. Autophagy induced by silica nanoparticles protects RAW264.7 macrophages from cell death.

    Science.gov (United States)

    Marquardt, Clarissa; Fritsch-Decker, Susanne; Al-Rawi, Marco; Diabaté, Silvia; Weiss, Carsten

    2017-03-15

    Although the technological and economic benefits of engineered nanomaterials are obvious, concerns have been raised about adverse effects if such material is inhaled, ingested, applied to the skin or even released into the environment. Here we studied the cytotoxic effects of the most abundant nanomaterial, silica nanoparticles (SiO 2 -NPs), in murine RAW264.7 macrophages. SiO 2 -NPs dose-dependently induce membrane leakage and cell death without obvious involvement of reactive oxygen species. Interestingly, at low concentrations SiO 2 -NPs trigger autophagy, evidenced by morphological and biochemical hallmarks such as autophagolysosomes or increased levels of LC3-II, which serves to protect cells from cytotoxicity. Hence SiO 2 -NPs initiate an adaptive stress response which dependent on dose serve to balance survival and death and ultimately dictates the cellular fate. Copyright © 2017 Elsevier B.V. All rights reserved.

  14. H.264/AVC digital fingerprinting based on spatio-temporal just noticeable distortion

    Science.gov (United States)

    Ait Saadi, Karima; Bouridane, Ahmed; Guessoum, Abderrezak

    2014-01-01

    This paper presents a robust adaptive embedding scheme using a modified Spatio-Temporal noticeable distortion (JND) model that is designed for tracing the distribution of the H.264/AVC video content and protecting them from unauthorized redistribution. The Embedding process is performed during coding process in selected macroblocks type Intra 4x4 within I-Frame. The method uses spread-spectrum technique in order to obtain robustness against collusion attacks and the JND model to dynamically adjust the embedding strength and control the energy of the embedded fingerprints so as to ensure their imperceptibility. Linear and non linear collusion attacks are performed to show the robustness of the proposed technique against collusion attacks while maintaining visual quality unchanged.

  15. Multiple descriptions based on multirate coding for JPEG 2000 and H.264/AVC.

    Science.gov (United States)

    Tillo, Tammam; Baccaglini, Enrico; Olmo, Gabriella

    2010-07-01

    Multiple description coding (MDC) makes use of redundant representations of multimedia data to achieve resiliency. Descriptions should be generated so that the quality obtained when decoding a subset of them only depends on their number and not on the particular received subset. In this paper, we propose a method based on the principle of encoding the source at several rates, and properly blending the data encoded at different rates to generate the descriptions. The aim is to achieve efficient redundancy exploitation, and easy adaptation to different network scenarios by means of fine tuning of the encoder parameters. We apply this principle to both JPEG 2000 images and H.264/AVC video data. We consider as the reference scenario the distribution of contents on application-layer overlays with multiple-tree topology. The experimental results reveal that our method favorably compares with state-of-art MDC techniques.

  16. A robust H.264/AVC video watermarking scheme with drift compensation.

    Science.gov (United States)

    Jiang, Xinghao; Sun, Tanfeng; Zhou, Yue; Wang, Wan; Shi, Yun-Qing

    2014-01-01

    A robust H.264/AVC video watermarking scheme for copyright protection with self-adaptive drift compensation is proposed. In our scheme, motion vector residuals of macroblocks with the smallest partition size are selected to hide copyright information in order to hold visual impact and distortion drift to a minimum. Drift compensation is also implemented to reduce the influence of watermark to the most extent. Besides, discrete cosine transform (DCT) with energy compact property is applied to the motion vector residual group, which can ensure robustness against intentional attacks. According to the experimental results, this scheme gains excellent imperceptibility and low bit-rate increase. Malicious attacks with different quantization parameters (QPs) or motion estimation algorithms can be resisted efficiently, with 80% accuracy on average after lossy compression.

  17. Adaptive quantization-parameter clip scheme for smooth quality in H.264/AVC.

    Science.gov (United States)

    Hu, Sudeng; Wang, Hanli; Kwong, Sam

    2012-04-01

    In this paper, we investigate the issues over the smooth quality and the smooth bit rate during rate control (RC) in H.264/AVC. An adaptive quantization-parameter (Q(p)) clip scheme is proposed to optimize the quality smoothness while keeping the bit-rate fluctuation at an acceptable level. First, the frame complexity variation is studied by defining a complexity ratio between two nearby frames. Second, the range of the generated bits is analyzed to prevent the encoder buffer from overflow and underflow. Third, based on the safe range of the generated bits, an optimal Q(p) clip range is developed to reduce the quality fluctuation. Experimental results demonstrate that the proposed Q(p) clip scheme can achieve excellent performance in quality smoothness and buffer regulation.

  18. Temporal Scalability through Adaptive -Band Filter Banks for Robust H.264/MPEG-4 AVC Video Coding

    Directory of Open Access Journals (Sweden)

    Pau G

    2006-01-01

    Full Text Available This paper presents different structures that use adaptive -band hierarchical filter banks for temporal scalability. Open-loop and closed-loop configurations are introduced and illustrated using existing video codecs. In particular, it is shown that the H.264/MPEG-4 AVC codec allows us to introduce scalability by frame shuffling operations, thus keeping backward compatibility with the standard. The large set of shuffling patterns introduced here can be exploited to adapt the encoding process to the video content features, as well as to the user equipment and transmission channel characteristics. Furthermore, simulation results show that this scalability is obtained with no degradation in terms of subjective and objective quality in error-free environments, while in error-prone channels the scalable versions provide increased robustness.

  19. Lipopolysaccharide induces multinuclear cell from RAW264.7 line with increased phagocytosis activity

    Energy Technology Data Exchange (ETDEWEB)

    Nakanishi-Matsui, Mayumi, E-mail: nakanim@iwate-med.ac.jp [Department of Biochemistry, Faculty of Pharmaceutical Sciences, Iwate Medical University, Futai Special Laboratory, Yahaba, Iwate 028-3694 (Japan); Yano, Shio; Matsumoto, Naomi; Futai, Masamitsu [Department of Biochemistry, Faculty of Pharmaceutical Sciences, Iwate Medical University, Futai Special Laboratory, Yahaba, Iwate 028-3694 (Japan)

    2012-08-24

    Highlights: Black-Right-Pointing-Pointer LPS induces multinuclear cells from murine macrophage-derived RAW264.7 cells. Black-Right-Pointing-Pointer The multinuclear cells are formed through cell-cell fusion in the presence of Ca{sup 2+}. Black-Right-Pointing-Pointer The multinuclear cells do not express osteoclast-specific enzymes. Black-Right-Pointing-Pointer They internalized more and larger beads than mononuclear cells and osteoclasts. -- Abstract: Lipopolysaccharide (LPS), an outer membrane component of Gram-negative bacteria, induces strong proinflammatory responses, including the release of cytokines and nitric oxide from macrophage. In this study, we found that a murine macrophage-derived line, RAW264.7, became multinuclear through cell-cell fusion after incubation with highly purified LPS or synthetic lipid A in the presence of Ca{sup 2+}. The same cell line is known to differentiate into multinuclear osteoclast, which expresses a specific proton pumping ATPase together with osteoclast markers on stimulation by the extracellular domain of receptor activator of nuclear factor {kappa}B ligand (Toyomura, T., Murata, Y., Yamamoto, A., Oka, T., Sun-Wada, G.-H., Wada, Y. and Futai, M., 2003). The LPS-induced multinuclear cells did not express osteoclast-specific enzymes including tartrate-resistant acid phosphatase and cathepsin K. During multinuclear cell formation, the cells internalized more and larger polystyrene beads (diameter 6-15 {mu}m) than mononuclear cells and osteoclasts. The internalized beads were located in lysosome-marker positive organelles, which were probably phagolysosomes. The LPS-induced multinuclear cell could be a good model system to study phagocytosis of large foreign bodies.

  20. Toxicity and oxidative stress induced by semiconducting polymer dots in RAW264.7 mouse macrophages

    Science.gov (United States)

    Ye, Fangmao; White, Collin C.; Jin, Yuhui; Hu, Xiaoge; Hayden, Sarah; Zhang, Xuanjun; Gao, Xiaohu; Kavanagh, Terrance J.; Chiu, Daniel T.

    2015-05-01

    The rapid development and acceptance of PDots for biological applications depends on an in depth understanding of their cytotoxicity. In this paper, we performed a comprehensive study of PDot cytotoxicity at both the gross cell effect level (such as cell viability, proliferation and necrosis) and more subtle effects (such as redox stress) on RAW264.7 cells, a murine macrophage cell line with high relevance to in vivo nanoparticle disposition. The redox stress measurements assessed were inner mitochondrial membrane lipid peroxidation (nonyl-acridine orange, NAO), total thiol level (monobromobimane, MBB), and pyridine nucleotide redox status (NAD(P)H autofluorescence). Because of the extensive work already performed with QDots on nanotoxicity and also because of their comparable size, QDots were chosen as a comparison/reference nanoparticle for this study. The results showed that PDots exhibit cytotoxic effects to a much lesser degree than their inorganic analogue (QDots) and are much brighter, allowing for much lower concentrations to be used in various biological applications. In addition, at lower dose levels (2.5 nM to 10 nM) PDot treatment resulted in higher total thiol level than those found with QDots. At higher dose levels (20 nM to 40 nM) QDots caused significantly higher thiol levels in RAW264.7 cells, than was seen with PDots, suggesting that QDots elicit compensation to oxidative stress by upregulating GSH synthesis. At the higher concentrations of QDots, NAD(P)H levels showed an initial depletion, then repletion to a level that was greater than vehicle controls. PDots showed a similar trend but this was not statistically significant. Because PDots elicit less oxidative stress and cytotoxicity at low concentrations than QDots, and because they exhibit superior fluorescence at these low concentrations, PDots are predicted to have enhanced utility in biomedical applications.

  1. [Etiological analysis of 264 cases with chronic kidney disease stage 2 to 5 in children].

    Science.gov (United States)

    Miao, Qianfan; Shen, Qian; Xu, Hong; Sun, Li; Tang, Xiaoshan; Fang, Xiaoyan; Liu, Haimei; Zhai, Yihui; Bi, Yunli; Wang, Xiang; Chen, Hong

    2015-09-01

    To study and summarize the etiology of children patients with chronic kidney disease (CKD) stage 2 to 5 seen in Children's Hospital of Fudan University from Jan. 2004 to Dec. 2013. By complying with the NKF-K/DOQI guidelines, we collected data of 264 cases of children patients with CKD stage 2-5 from Jan. 2004 to Dec. 2013 in the medical record system of Children's Hospital of Fudan University. And we retrospectively analyzed their age and CKD stage at first diagnosis, primary diseases, complications, etc. In the collected 264 cases, 52 cases (19.7%) were diagnosed at stage 2, 67 (25.4%) at stage 3, 52 (19.7%) at stage 4 and 93 (35.2%) at stage 5. For disease causes, 116 cases (43.9%) had congenital anomalies of the kidney and urinary tract (CAKUT), 61 cases (23.1%) had glomerular disease, 15 (5.7%) had hereditary kidney disease, 14 (5.3%) had other diseases and in 58 cases (22.0%) the causes of disease were unknown. In the group with age between 0 and 3.0 and 3.1 and 6.0 years, 57.1% (24 cases) and 60.0% (30 cases) had primary disease with CAKUT. In the group with age older than 10 years, 49.2% (30 cases) had primary disease with glomerular disease and 32.0% (32 cases) with unknown causes. The major cause of CKD stage 2-5 in children in our hospital during the last ten years was CAKUT (43.9%), followed by glomerular disease (23.1%). The primary diseases of CKD were significantly different between the 2 age groups. CAKUT was more common in infants and preschool children while for adolescents, glomerular disease was the major cause.

  2. WebM een alternatief voor H.264? : Onderzoek naar het gebruik van open source software en open standaarden

    NARCIS (Netherlands)

    Staalduinen, M. van; Prins, M.J.

    2011-01-01

    WebM is een nieuw videoformaat en wordt gepositioneerd als een open en vrij te gebruiken alternatief voor de videostandaard H.264. Deze standaard wordt op dit moment het meest gebruikt onder andere voor Uitzending Gemist. Deze formaten zijn regelmatig met elkaar vergeleken, waar veelal wordt

  3. Complexity Control of Fast Motion Estimation in H.264/MPEG-4 AVC with Rate-Distortion-Complexity optimization

    DEFF Research Database (Denmark)

    Wu, Mo; Forchhammer, Søren; Aghito, Shankar Manuel

    2007-01-01

    A complexity control algorithm for H.264 advanced video coding is proposed. The algorithm can control the complexity of integer inter motion estimation for a given target complexity. The Rate-Distortion-Complexity performance is improved by a complexity prediction model, simple analysis of the pa...

  4. 12 CFR 264.101 - Cross-reference to employees' ethical conduct standards and financial disclosure regulations.

    Science.gov (United States)

    2010-01-01

    ... 12 Banks and Banking 3 2010-01-01 2010-01-01 false Cross-reference to employees' ethical conduct... § 264.101 Cross-reference to employees' ethical conduct standards and financial disclosure regulations... branch-wide standards of ethical conduct at 5 CFR part 2635 and the Board's regulation at 5 CFR part 6801...

  5. A large-scale video codec comparison of x264, x265 and libvpx for practical VOD applications

    Science.gov (United States)

    De Cock, Jan; Mavlankar, Aditya; Moorthy, Anush; Aaron, Anne

    2016-09-01

    Over the last years, we have seen exciting improvements in video compression technology, due to the introduction of HEVC and royalty-free coding specifications such as VP9. The potential compression gains of HEVC over H.264/AVC have been demonstrated in different studies, and are usually based on the HM reference software. For VP9, substantial gains over H.264/AVC have been reported in some publications, whereas others reported less optimistic results. Differences in configurations between these publications make it more difficult to assess the true potential of VP9. Practical open-source encoder implementations such as x265 and libvpx (VP9) have matured, and are now showing high compression gains over x264. In this paper, we demonstrate the potential of these encoder imple- mentations, with settings optimized for non-real-time random access, as used in a video-on-demand encoding pipeline. We report results from a large-scale video codec comparison test, which includes x264, x265 and libvpx. A test set consisting of a variety of titles with varying spatio-temporal characteristics from our catalog is used, resulting in tens of millions of encoded frames, hence larger than test sets previously used in the literature. Re- sults are reported in terms of PSNR, SSIM, MS-SSIM, VIF and the recently introduced VMAF quality metric. BD-rate calculations show that using x265 and libvpx vs. x264 can lead to significant bitrate savings for the same quality. x265 outperforms libvpx in most cases, but the performance gap narrows (or even reverses) at the higher resolutions.

  6. Characterization of CYP264B1 and a terpene cyclase of a terpene biosynthesis gene cluster from the myxobacterium Sorangium cellulosum So ce56

    OpenAIRE

    Ly, Thuy Thi Bich

    2011-01-01

    In the work presented here, CYP264B1 and the terpene cyclase GeoA of Sorangium cellulosum So ce56 have been characterized. CYP264B1 is able to convert norisoprenoids (a-ionone and b-ionone) and diverse sesquiterpene compounds, including nootkatone. Three products, 3-hydroxy-a-ionone, 3-hydroxy-b-ionone and 13-hydroxy-nootkatone were characterized using HPLC and 1H and 13C NMR. CYP264B1 is the first enzyme reported to be capable to hydroxylate regioselectively both norisoprenoids at the positi...

  7. Graceful Degradation in 3GPP MBMS Mobile TV Services Using H.264/AVC Temporal Scalability

    Directory of Open Access Journals (Sweden)

    Thomas Wiegand

    2009-01-01

    Full Text Available These days, there is an increasing interest in Mobile TV broadcast services shown by customers as well as service providers. One general problem of Mobile TV broadcast services is to maximize the coverage of users receiving an acceptable service quality, which is mainly influenced by the user's position and mobility within the cell. In this paper, graceful degradation is considered as an approach for improved service availability and coverage. We present a layered transmission approach for 3GPP's Release 6 Multimedia and Broadcast Service (MBMS based on temporal scalability using H.264/AVC Baseline Profile. A differentiation in robustness between temporal quality layers is achieved by unequal error protection approach based on either application layer Forward Error Correction (FEC or unequal transmit power for the layers or even a combination of both. We discuss the corresponding MBMS service as well as network settings and define measures allowing for evaluating the amount of users reached with a certain mobile terminal play-out quality while considering the network cell capacity usage. Using simulated 3GPP Rel. 6 network conditions, we show that if the service and network settings are chosen carefully, a noticeable extension of the coverage of the MBMS service can be achieved.

  8. Adaptive order search and tangent-weighted trade-off for motion estimation in H.264

    Directory of Open Access Journals (Sweden)

    Srinivas Bachu

    2018-04-01

    Full Text Available Motion estimation and compensation play a major role in video compression to reduce the temporal redundancies of the input videos. A variety of block search patterns have been developed for matching the blocks with reduced computational complexity, without affecting the visual quality. In this paper, block motion estimation is achieved through integrating the square as well as the hexagonal search patterns with adaptive order. The proposed algorithm is called, AOSH (Adaptive Order Square Hexagonal Search algorithm, and it finds the best matching block with a reduced number of search points. The searching function is formulated as a trade-off criterion here. Hence, the tangent-weighted function is newly developed to evaluate the matching point. The proposed AOSH search algorithm and the tangent-weighted trade-off criterion are effectively applied to the block estimation process to enhance the visual quality and the compression performance. The proposed method is validated using three videos namely, football, garden and tennis. The quantitative performance of the proposed method and the existing methods is analysed using the Structural SImilarity Index (SSIM and the Peak Signal to Noise Ratio (PSNR. The results prove that the proposed method offers good visual quality than the existing methods. Keywords: Block motion estimation, Square search, Hexagon search, H.264, Video coding

  9. EFFICIENT BLOCK MATCHING ALGORITHMS FOR MOTION ESTIMATION IN H.264/AVC

    Directory of Open Access Journals (Sweden)

    P. Muralidhar

    2015-02-01

    Full Text Available In Scalable Video Coding (SVC, motion estimation and inter-layer prediction play an important role in elimination of temporal and spatial redundancies between consecutive layers. This paper evaluates the performance of widely accepted block matching algorithms used in various video compression standards, with emphasis on the performance of the algorithms for a didactic scalable video codec. Many different implementations of Fast Motion Estimation Algorithms have been proposed to reduce motion estimation complexity. The block matching algorithms have been analyzed with emphasis on Peak Signal to Noise Ratio (PSNR and computations using MATLAB. In addition to the above comparisons, a survey has been done on Spiral Search Motion Estimation Algorithms for Video Coding. A New Modified Spiral Search (NMSS motion estimation algorithm has been proposed with lower computational complexity. The proposed algorithm achieves 72% reduction in computation with a minimal (<1dB reduction in PSNR. A brief introduction to the entire flow of video compression H.264/SVC is also presented in this paper.

  10. WATERLOPP V2/64: A highly parallel machine for numerical computation

    Science.gov (United States)

    Ostlund, Neil S.

    1985-07-01

    Current technological trends suggest that the high performance scientific machines of the future are very likely to consist of a large number (greater than 1024) of processors connected and communicating with each other in some as yet undetermined manner. Such an assembly of processors should behave as a single machine in obtaining numerical solutions to scientific problems. However, the appropriate way of organizing both the hardware and software of such an assembly of processors is an unsolved and active area of research. It is particularly important to minimize the organizational overhead of interprocessor comunication, global synchronization, and contention for shared resources if the performance of a large number ( n) of processors is to be anything like the desirable n times the performance of a single processor. In many situations, adding a processor actually decreases the performance of the overall system since the extra organizational overhead is larger than the extra processing power added. The systolic loop architecture is a new multiple processor architecture which attemps at a solution to the problem of how to organize a large number of asynchronous processors into an effective computational system while minimizing the organizational overhead. This paper gives a brief overview of the basic systolic loop architecture, systolic loop algorithms for numerical computation, and a 64-processor implementation of the architecture, WATERLOOP V2/64, that is being used as a testbed for exploring the hardware, software, and algorithmic aspects of the architecture.

  11. Clinical Significance of Internal Mammary Lymph Node Biopsy during Microsurgical Breast Reconstruction: Review of 264 Cases.

    Science.gov (United States)

    Wright, Eric J; Momeni, Arash; Kraneburg, Ursula M; Otake, Leo R; Echo, Anthony; Lee, Tim; Buchanan, Edward P; Lee, Gordon K

    2016-06-01

    Despite the knowledge of alternate lymphatic draining patterns of the breast, routine evaluation of the internal mammary lymph node basin is still not considered standard of care. The advent of microsurgical breast reconstruction using the internal mammary vessels as recipients, however, has allowed sampling of internal mammary lymph nodes with technical ease, thus revisiting their role in breast cancer management. In the present study, the authors reviewed their experience with this practice. A retrospective analysis of patients who underwent internal mammary lymph node biopsy at the time of autologous breast reconstruction using the internal mammary vessels between 2004 and 2012 was performed. Parameters of interest included patient age, timing of reconstruction (immediate versus delayed), disease stage, and pathologic findings of internal mammary lymph nodes. A total of 264 autologous breast reconstructions using the internal mammary vessels were performed in 204 patients with a median age of 44.5 years. The majority of reconstructions were immediate [n = 211 (79.9 percent)]. Seventy-two percent of patients had either stage I [72 patients (35.3 percent)] or stage II disease [75 patients (36.8 percent)]. Six patients were found to have internal mammary lymph node metastasis. Stage migration and alteration in adjuvant therapy occurred in all patients. Internal mammary lymph node sampling at the time of autologous breast reconstruction using the internal mammary system should become routine practice, as the morbidity associated with internal mammary lymph node harvest is low and the impact in cases of nodal involvement is quite substantial. Therapeutic, IV.

  12. Lipopolysaccharide induced MAP kinase activation in RAW 264.7 cells attenuated by cerium oxide nanoparticles

    Directory of Open Access Journals (Sweden)

    Vellaisamy Selvaraj

    2015-09-01

    Full Text Available High mortality rates are associated with the life threatening disease of sepsis. Improvements in septic patient survivability have failed to materialize with currently available treatments. This article represents data regarding a study published in biomaterials (Vellaisamy et al., Biomaterials, 2015, in press. with the purpose of evaluating whether severe sepsis mortality and associated hepatic dysfunction induced by lipopolysaccharide (LPS can be prevented by cerium oxide nanoparticles (CeO2NPs treatment in male Sprague Dawley rats. Here we provide the information about the method and processing of raw data related to our study publish in Biomaterials and Data in Brief (Vellaisamy et al., Biomaterials, 2015, in press; Vellaisamy et al., Data in Brief, 2015, in press.. The data contained in this article evaluates the contribution of MAPK signaling in LPS induced sepsis. Macrophage cells (RAW 264.7 were treated with a range of cerium oxide nanoparticle concentration in the presence and absence of LPS. Immunoblotting was performed on the cell lysates to evaluate the effect of cerium oxide nanoparticle treatment on LPS induced changes in Mitogen Activated Protein Kinases (MAPK p-38, ERK 1/2, and SAPK/JNK phosphorylation.

  13. A CABAC codec of H.264AVC with secure arithmetic coding

    Science.gov (United States)

    Neji, Nihel; Jridi, Maher; Alfalou, Ayman; Masmoudi, Nouri

    2013-02-01

    This paper presents an optimized H.264/AVC coding system for HDTV displays based on a typical flow with high coding efficiency and statics adaptivity features. For high quality streaming, the codec uses a Binary Arithmetic Encoding/Decoding algorithm with high complexity and a JVCE (Joint Video compression and encryption) scheme. In fact, particular attention is given to simultaneous compression and encryption applications to gain security without compromising the speed of transactions [1]. The proposed design allows us to encrypt the information using a pseudo-random number generator (PRNG). Thus we achieved the two operations (compression and encryption) simultaneously and in a dependent manner which is a novelty in this kind of architecture. Moreover, we investigated the hardware implementation of CABAC (Context-based adaptive Binary Arithmetic Coding) codec. The proposed architecture is based on optimized binarizer/de-binarizer to handle significant pixel rates videos with low cost and high performance for most frequent SEs. This was checked using HD video frames. The obtained synthesis results using an FPGA (Xilinx's ISE) show that our design is relevant to code main profile video stream.

  14. Rab20 regulates phagosome maturation in RAW264 macrophages during Fc gamma receptor-mediated phagocytosis.

    Directory of Open Access Journals (Sweden)

    Youhei Egami

    Full Text Available Rab20, a member of the Rab GTPase family, is known to be involved in membrane trafficking, however its implication in FcγR-mediated phagocytosis is unclear. We examined the spatiotemporal localization of Rab20 during phagocytosis of IgG-opsonized erythrocytes (IgG-Es in RAW264 macrophages. By the live-cell imaging of fluorescent protein-fused Rab20, it was shown that Rab20 was transiently associated with the phagosomal membranes. During the early stage of phagosome formation, Rab20 was not localized on the membranes of phagocytic cups, but was gradually recruited to the newly formed phagosomes. Although Rab20 was colocalized with Rab5 to some extent, the association of Rab20 with the phagosomes persisted even after the loss of Rab5 from the phagosomal membranes. Then, Rab20 was colocalized with Rab7 and Lamp1, late endosomal/lysosomal markers, on the internalized phagosomes. Moreover, our analysis of Rab20 mutant expression revealed that the maturation of phagosomes was significantly delayed in cells expressing the GDP-bound mutant Rab20-T19N. These data suggest that Rab20 is an important component of phagosome and regulates the phagosome maturation during FcγR-mediated phagocytosis.

  15. Optimized bit extraction using distortion modeling in the scalable extension of H.264/AVC.

    Science.gov (United States)

    Maani, Ehsan; Katsaggelos, Aggelos K

    2009-09-01

    The newly adopted scalable extension of H.264/AVC video coding standard (SVC) demonstrates significant improvements in coding efficiency in addition to an increased degree of supported scalability relative to the scalable profiles of prior video coding standards. Due to the complicated hierarchical prediction structure of the SVC and the concept of key pictures, content-aware rate adaptation of SVC bit streams to intermediate bit rates is a nontrivial task. The concept of quality layers has been introduced in the design of the SVC to allow for fast content-aware prioritized rate adaptation. However, existing quality layer assignment methods are suboptimal and do not consider all network abstraction layer (NAL) units from different layers for the optimization. In this paper, we first propose a technique to accurately and efficiently estimate the quality degradation resulting from discarding an arbitrary number of NAL units from multiple layers of a bitstream by properly taking drift into account. Then, we utilize this distortion estimation technique to assign quality layers to NAL units for a more efficient extraction. Experimental results show that a significant gain can be achieved by the proposed scheme.

  16. Network-based H.264/AVC whole frame loss visibility model and frame dropping methods.

    Science.gov (United States)

    Chang, Yueh-Lun; Lin, Ting-Lan; Cosman, Pamela C

    2012-08-01

    We examine the visual effect of whole frame loss by different decoders. Whole frame losses are introduced in H.264/AVC compressed videos which are then decoded by two different decoders with different common concealment effects: frame copy and frame interpolation. The videos are seen by human observers who respond to each glitch they spot. We found that about 39% of whole frame losses of B frames are not observed by any of the subjects, and over 58% of the B frame losses are observed by 20% or fewer of the subjects. Using simple predictive features which can be calculated inside a network node with no access to the original video and no pixel level reconstruction of the frame, we developed models which can predict the visibility of whole B frame losses. The models are then used in a router to predict the visual impact of a frame loss and perform intelligent frame dropping to relieve network congestion. Dropping frames based on their visual scores proves superior to random dropping of B frames.

  17. Minimum variance optimal rate allocation for multiplexed H.264/AVC bitstreams.

    Science.gov (United States)

    Tagliasacchi, Marco; Valenzise, Giuseppe; Tubaro, Stefano

    2008-07-01

    Consider the problem of transmitting multiple video streams to fulfill a constant bandwidth constraint. The available bit budget needs to be distributed across the sequences in order to meet some optimality criteria. For example, one might want to minimize the average distortion or, alternatively, minimize the distortion variance, in order to keep almost constant quality among the encoded sequences. By working in the rho-domain, we propose a low-delay rate allocation scheme that, at each time instant, provides a closed form solution for either the aforementioned problems. We show that minimizing the distortion variance instead of the average distortion leads, for each of the multiplexed sequences, to a coding penalty less than 0.5 dB, in terms of average PSNR. In addition, our analysis provides an explicit relationship between model parameters and this loss. In order to smooth the distortion also along time, we accommodate a shared encoder buffer to compensate for rate fluctuations. Although the proposed scheme is general, and it can be adopted for any video and image coding standard, we provide experimental evidence by transcoding bitstreams encoded using the state-of-the-art H.264/AVC standard. The results of our simulations reveal that is it possible to achieve distortion smoothing both in time and across the sequences, without sacrificing coding efficiency.

  18. Fast bi-directional prediction selection in H.264/MPEG-4 AVC temporal scalable video coding.

    Science.gov (United States)

    Lin, Hung-Chih; Hang, Hsueh-Ming; Peng, Wen-Hsiao

    2011-12-01

    In this paper, we propose a fast algorithm that efficiently selects the temporal prediction type for the dyadic hierarchical-B prediction structure in the H.264/MPEG-4 temporal scalable video coding (SVC). We make use of the strong correlations in prediction type inheritance to eliminate the superfluous computations for the bi-directional (BI) prediction in the finer partitions, 16×8/8×16/8×8 , by referring to the best temporal prediction type of 16 × 16. In addition, we carefully examine the relationship in motion bit-rate costs and distortions between the BI and the uni-directional temporal prediction types. As a result, we construct a set of adaptive thresholds to remove the unnecessary BI calculations. Moreover, for the block partitions smaller than 8 × 8, either the forward prediction (FW) or the backward prediction (BW) is skipped based upon the information of their 8 × 8 partitions. Hence, the proposed schemes can efficiently reduce the extensive computational burden in calculating the BI prediction. As compared to the JSVM 9.11 software, our method saves the encoding time from 48% to 67% for a large variety of test videos over a wide range of coding bit-rates and has only a minor coding performance loss. © 2011 IEEE

  19. Effect of different combinations of MGK-264 or piperonyl butoxide with plant-derived molluscicides on snail reproduction.

    Science.gov (United States)

    Singh, K; Singh, D K

    2000-02-01

    Effect of sublethal treatment (20% and 60% of LC(50)/24 h) of plant-derived molluscicides, viz. Polianthes tuberosa, Trachyspermum ammi, Allium sativum powder; Azadirachta indica oil; oleoresin of Zingiber officinale; and their active molluscicidal component in combination (1:5) with MGK-264 or piperonyl butoxide on the reproduction of snail Lymnaea acuminata have been studied. It was observed that the combination of plant derived molluscicide and their active molluscicidal components, viz. tigogenin, hecogenin, azadirachtin, allicin, thymol, and [6]-gingerol combination with MGK-264 or piperonyl butoxide caused a significant reduction in fecundity, hatchability, and survival of young snails. Withdrawal of snails to fresh water after the above treatment caused a significant recovery in the fecundity of L. acuminata.

  20. Inhibitory Effects of Soyeum Pharmacopuncture (SPP on LPS-induced Inflammation Related Cytokine Expressions of RAW 264.7 cells

    Directory of Open Access Journals (Sweden)

    Yoon Mi-Young

    2007-12-01

    Full Text Available Aim : This study was done to investigate whether SPP has inhibitory effects on the activation of RAW 264.7 cells. Method : In tumor necrosis factor-a (TNF-a/ interleukin-1b (IL-1b and IL-6, the mRNA expression of molecular indicators related to inflammatory changes of the Reumatoid Arthritis (RA were examined using quantitative real-time PCR. Results : The treatment of SPP significantly suppressed the expression of proinflammatory cytokines and chemokines such as TNF-a, IL-1b, IL-6 compared with the control. The expression of NOS-II was considerably reduced, which was accompanied by a reduction in the production of nitric oxide (NO. It also reduced the expression of TNF-αin serum of Balb/c mice compared with control group. Conclusion : SPP is an effective herbal material for suppressing the inflammation related cytokines of RAW 264.7 cells.

  1. Impact of H.264/AVC and H.265/HEVC Compression Standards on the Video Quality for 4K Resolution

    Directory of Open Access Journals (Sweden)

    Miroslav Uhrina

    2014-01-01

    Full Text Available This article deals with the impact of H.264/AVC and H.265/HEVC compression standards on the video quality for 4K resolution. In the first part a short characteristic of both compression standards is written. The second part focuses on the well-known objective metrics which were used for evaluating the video quality. In the third part the measurements and the experimental results are described.

  2. Anti-Inflammatory Phenolic Metabolites from the Edible Fungus Phellinus baumii in LPS-Stimulated RAW264.7 Cells

    Directory of Open Access Journals (Sweden)

    Seulah Lee

    2017-09-01

    Full Text Available The edible fungus Phellinus baumii Pilat (Hymenochaetaceae has been used in Korean traditional medicines for strengthening health and prolonging life. An extract of the fruiting bodies of P. baumii was subjected to bioassay-guided fractionation based on its anti-inflammatory effects in lipopolysaccharide (LPS-stimulated RAW264.7 cells. The resulting fractions were chemically investigated, leading to isolation of three phenolic compounds (1–3, a sesquiterpene (4, two steroids (5–6, a fatty acid (7, and a cerebroside (8. Spectroscopic analyses including 1D and 2D NMR spectroscopy and LC/MS were used to determine their chemical structures. Compounds 2, 4, 5, 7 and 8 were identified in P. baumii for the first time. Since all compounds were isolated from active fractions with anti-inflammatory activity, their ability to inhibit LPS-stimulated nitric oxide (NO production in RAW264.7 cells were evaluated in vitro. Compounds 1, 2, 3, 5 and 7 inhibited LPS-stimulated NO production, and compounds 1–3 had IC50 values <10 μM. Treatment of LPS-stimulated RAW264.7 cells with compounds 1–3 inhibited phosphorylation of IKKα and IκBα. In addition, treatment of compounds 1–3 reduced LPS-induced increases of nuclear factor-kappa B (NF-κB p65, iNOS and COX-2 protein expressions. Collectively, compounds 1–3 inhibited NF-κB-dependent inflammation in RAW264.7 cells. Thus, P. baumii is a potential source of natural anti-inflammatory agents, and active compounds 1–3 could be promising lead compounds for the development of novel anti-inflammatory agents.

  3. The optimization of H.264/AVC baseline decoder on low-cost TriMedia DSP processor

    Science.gov (United States)

    Wang, Sung-Wen; Yang, Ya-Ting; Li, Chia-Ying; Tung, Yi-Shin; Wu, Ja-Ling

    2004-11-01

    The emerging video coding standard, H.264/AVC, exhibits the unprecedented coding performance. Comparing to traditional coders, e.g., MPEG-2 and MEPG-4 ASP, about half bitrate saving is shown in the official verification test. Such outstanding performance makes it become the video compression candidate for the upcoming HD-DVD. As a side effect, it was also blamed that H.264/AVC is much more logically complex and requires more computation power than any of the existing standards. A low-cost and efficient implementation of the international standard hence plays an important role of its success. In this paper, we realize an H.264/AVC baseline decoder by a low-cost DSP processor, i.e., Philips" TriMedia TM-1300, and illustrate that less computation demand for H.264/AVC decoding becomes feasible by using effective software core. To this end, we first consider different approaches and take advantage of SIMD instruction set to optimize critical time-consuming coding modules, such as the fractional motion compensation, spatial prediction and inverse transform. Next, we also present some other optimization approaches for entropy decoding and in-loop deblocking filtering, even though they cannot get benefits from utilizing SIMD. In our experiments, by exploiting appropriate instruction level parallelism and efficient algorithms, the decoding speed can be improved by a factor of 8~10; a CIF video sequence can be decoded at up to 19.74~28.97 fps on a 166-MHz TriMedia TM-1300 processor compared to 2.40~2.98 fps by the standard reference software.

  4. Regulation of the efflux of putrescine and cadaverine from rapidly growing cultured RAW 264 cells by extracellular putrescine.

    OpenAIRE

    Tjandrawinata, R R; Byus, C V

    1995-01-01

    Cultures of the macrophage-like RAW 264 cells were adapted to divide normally in a synthetic serum-supplemented culture medium lacking any polyamines and diamine oxidase activity. These rapidly dividing cells actively effluxed large amounts of putrescine and cadaverine, compared with the intracellular levels, into the culture medium. The efflux of putrescine was stimulated by the amino acid ornithine, whereas efflux of cadaverine was inhibited. Relatively low levels of spermidine and N1-acety...

  5. The effects of Lycii Radicis Cortex on RANKL-induced osteoclast differentiation and activation in RAW 264.7 cells

    OpenAIRE

    KIM, JAE-HYUN; KIM, EUN-YOUNG; LEE, BINA; MIN, JU-HEE; SONG, DEA-UK; LIM, JEONG-MIN; EOM, JI WHAN; YEOM, MIJUNG; JUNG, HYUK-SANG; SOHN, YOUNGJOO

    2016-01-01

    Post-menopausal osteoporosis is a serious age-related disease. After the menopause, estrogen deficiency is common, and excessive osteoclast activity causes osteoporosis. Osteoclasts are multinucleated cells generated from the differentiation of monocyte/macrophage precursor cells such as RAW 264.7 cells. The water extract of Lycii Radicis Cortex (LRC) is made from the dried root bark of Lycium chinense Mill. and is termed 'Jigolpi' in Korea. Its effects on osteoclastogenesis and post-menopaus...

  6. Video coding standards AVS China, H.264/MPEG-4 PART 10, HEVC, VP6, DIRAC and VC-1

    CERN Document Server

    Rao, K R; Hwang, Jae Jeong

    2014-01-01

    Review by Ashraf A. Kassim, Professor, Department of Electrical & Computer Engineering, and Associate Dean, School of Engineering, National University of Singapore.     The book consists of eight chapters of which the first two provide an overview of various video & image coding standards, and video formats. The next four chapters present in detail the Audio & video standard (AVS) of China, the H.264/MPEG-4 Advanced video coding (AVC) standard, High efficiency video coding (HEVC) standard and the VP6 video coding standard (now VP10) respectively. The performance of the wavelet based Dirac video codec is compared with H.264/MPEG-4 AVC in chapter 7. Finally in chapter 8, the VC-1 video coding standard is presented together with VC-2 which is based on the intra frame coding of Dirac and an outline of a H.264/AVC to VC-1 transcoder.   The authors also present and discuss relevant research literature such as those which document improved methods & techniques, and also point to other related reso...

  7. An evaluation of the effect of JPEG, JPEG2000, and H.264/AVC on CQR codes decoding process

    Science.gov (United States)

    Vizcarra Melgar, Max E.; Farias, Mylène C. Q.; Zaghetto, Alexandre

    2015-02-01

    This paper presents a binarymatrix code based on QR Code (Quick Response Code), denoted as CQR Code (Colored Quick Response Code), and evaluates the effect of JPEG, JPEG2000 and H.264/AVC compression on the decoding process. The proposed CQR Code has three additional colors (red, green and blue), what enables twice as much storage capacity when compared to the traditional black and white QR Code. Using the Reed-Solomon error-correcting code, the CQR Code model has a theoretical correction capability of 38.41%. The goal of this paper is to evaluate the effect that degradations inserted by common image compression algorithms have on the decoding process. Results show that a successful decoding process can be achieved for compression rates up to 0.3877 bits/pixel, 0.1093 bits/pixel and 0.3808 bits/pixel for JPEG, JPEG2000 and H.264/AVC formats, respectively. The algorithm that presents the best performance is the H.264/AVC, followed by the JPEG2000, and JPEG.

  8. Low-magnitude high-frequency vibration inhibits RANKL-induced osteoclast differentiation of RAW264.7 cells.

    Science.gov (United States)

    Wu, Song-Hui; Zhong, Zhao-Ming; Chen, Jian-Ting

    2012-01-01

    Osteoclasts are the key participants in regulation of bone mass. Low-magnitude high-frequency vibration (LMHFV) has been found to be anabolic to bone in vivo. This study aimed to investigate the effect of LMHFV on osteoclast differentiation in vitro. Murine monocyte cell line RAW264.7 cells in the presence of receptor activator of nuclear factor-kappaB ligand (RANKL) were treated with or without LMHFV at 45 Hz (0.3 g) for 15 min day(-1). Tartrate resistant acid phosphatase (TRAP)-positive multinucleated cells (MNCs) and actin ring formation were evaluated. Expression of the osteoclast-specific genes, such as cathepsin K, matrix metallopeptidase-9 (MMP-9) and TRAP, were analyzed using real time-PCR. c-Fos, an osteoclast-specific transcription factor, was determined using Western blot. We found that LMHFV significantly decreased the number of RANKL-induced TRAP-positive MNCs (P<0.01), and inhibited the actin ring formation. The mRNA expression of the cathepsin K, MMP-9 and TRAP were down-regulated by LMHFV intervention (all P<0.001). Furthermore, LMHFV also inhibited the expression of c-Fos protein in the RANKL-treated RAW264.7 cells (P<0.05). Our results suggest that LMHFV can inhibit the RANKL-induced osteoclast differentiation of RAW264.7 cells, which give some new insight into the anabolic effects of LMHFV on bone.

  9. Anti-inflammatory effect of Capuli cherry against LPS-induced cytotoxic damage in RAW 264.7 macrophages.

    Science.gov (United States)

    Alvarez-Suarez, José M; Carrillo-Perdomo, Estefanía; Aller, Angel; Giampieri, Francesca; Gasparrini, Massimiliano; González-Pérez, Lien; Beltrán-Ayala, Pablo; Battino, Maurizio

    2017-04-01

    Capuli cherry (Prunus serotina Ehr. subsp. capuli (Cav.) McVaugh) fruits from the inter-Andean region of Ecuador were analysed to determine their bioactive compounds content, total antioxidant capacity, radical scavenging activity and their anti-inflammatory and protective effects against the cytotoxic damage mediated by lipopolysaccharide (LPS) in RAW 264.7 macrophages. Capuli fruits proved to be a natural source of bioactive compounds such as anthocyanins, vitamin C and β-carotene as well as to present an important total antioxidant capacity and radical scavenging activities. RAW 264.7 macrophages were incubated with different concentration of Capuli crude extract and subsequently activated by LPS to determine the markers related to oxidative damage and the proinflammatory cytokine production. The markers of oxidative damage, nitrite levels, the interleukin 1β messenger RNA levels and the tumor necrosis factor α mRNA levels and secretion were significantly decreased after the pre-incubated with Capuli extract and subsequently stimulated with LPS. In summary, Capuli extract attenuated the LPS-induced damage in RAW 264.7 macrophages due to its antioxidant and anti-inflammatory properties, showing that Capuli fruits may represent a relevant source of bioactive compounds with promising benefits for human health. Copyright © 2017 Elsevier Ltd. All rights reserved.

  10. Complexity reduction in the H.264/AVC using highly adaptive fast mode decision based on macroblock motion activity

    Science.gov (United States)

    Abdellah, Skoudarli; Mokhtar, Nibouche; Amina, Serir

    2015-11-01

    The H.264/AVC video coding standard is used in a wide range of applications from video conferencing to high-definition television according to its high compression efficiency. This efficiency is mainly acquired from the newly allowed prediction schemes including variable block modes. However, these schemes require a high complexity to select the optimal mode. Consequently, complexity reduction in the H.264/AVC encoder has recently become a very challenging task in the video compression domain, especially when implementing the encoder in real-time applications. Fast mode decision algorithms play an important role in reducing the overall complexity of the encoder. In this paper, we propose an adaptive fast intermode algorithm based on motion activity, temporal stationarity, and spatial homogeneity. This algorithm predicts the motion activity of the current macroblock from its neighboring blocks and identifies temporal stationary regions and spatially homogeneous regions using adaptive threshold values based on content video features. Extensive experimental work has been done in high profile, and results show that the proposed source-coding algorithm effectively reduces the computational complexity by 53.18% on average compared with the reference software encoder, while maintaining the high-coding efficiency of H.264/AVC by incurring only 0.097 dB in total peak signal-to-noise ratio and 0.228% increment on the total bit rate.

  11. Separation and purification of bioactive botrallin and TMC-264 by a combination of HSCCC and semi-preparative HPLC from endophytic fungus Hyalodendriella sp. Ponipodef12.

    Science.gov (United States)

    Mao, Ziling; Luo, Ruiya; Luo, Haiyu; Tian, Jin; Liu, Hongwei; Yue, Yang; Wang, Mingan; Peng, Youliang; Zhou, Ligang

    2014-09-01

    Two dibenzo-α-pyrones, botrallin (1) and TMC-264 (2) were preparatively separated from crude ethyl acetate extract of the endophytic fungus Hyalodendriella sp. Ponipodef12, which was isolated from the hybrid 'Neva' of Populus deltoides Marsh × P. nigra L. using a combination of high-speed counter-current chromatography (HSCCC) and semi-preparative HPLC. Botrallin (1) with 74.73% of purity and TMC-264 (2) with 82.29% of purity were obtained through HSCCC by employing a solvent system containing n-hexane-ethyl acetate-methanol-water at a volume ratio of 1.2:1.0:0.9:1.0. It was the first time for TMC-264 (2) to be isolated from this fungus. TMC-264 (2) showed strong antimicrobial and antinematodal activity, and botrallin (1) exhibited moderate inhibitory activity on acetylcholinesterase.

  12. Enhancement of botrallin and TMC-264 production in liquid culture of endophytic fungus Hyalodendriella sp. Ponipodef12 after treatments with metal ions

    Directory of Open Access Journals (Sweden)

    Haiyu Luo

    2016-11-01

    Conclusion: The results indicated that the enhancement of botrallin and TMC-264 accumulation in liquid culture of the endophytic fungus Hyalodendriella sp. Ponipodef12 by the metal ions and their combination should be an effective strategy.

  13. Double stranded RNA-dependent protein kinase is involved in osteoclast differentiation of RAW264.7 cells in vitro

    Energy Technology Data Exchange (ETDEWEB)

    Teramachi, Junpei [Department of Histology and Oral Histology, Institute of Health Biosciences, The University of Tokushima Graduate School, Kuramoto, Tokushima 770-8504 (Japan); Morimoto, Hiroyuki; Baba, Ryoko; Doi, Yoshiaki [Department of Anatomy, School of Medicine, University of Occupational and Environmental Health, Yahatanishi, Kitakyushu 807-8555 (Japan); Hirashima, Kanji [Department of Histology and Oral Histology, Institute of Health Biosciences, The University of Tokushima Graduate School, Kuramoto, Tokushima 770-8504 (Japan); Haneji, Tatsuji, E-mail: tat-hane@dent.tokushima-u.ac.jp [Department of Histology and Oral Histology, Institute of Health Biosciences, The University of Tokushima Graduate School, Kuramoto, Tokushima 770-8504 (Japan)

    2010-11-15

    Double-stranded RNA-dependent protein kinase (PKR) plays a critical role in antiviral defence of the host cells. PKR is also involved in cell cycle progression, cell proliferation, cell differentiation, tumorigenesis, and apoptosis. We previously reported that PKR is required for differentiation and calcification of osteoblasts. However, it is unknown about the role of PKR in osteoclast differentiation. A dominant-negative PKR mutant cDNA, in which the amino acid lysine at 296 was replaced with arginine, was transfected into RAW264.7 cells. We have established the cell line that stably expresses the PKR mutant gene (PKR-K/R). Phosphorylation of PKR and {alpha}-subunit of eukaryotic initiation factor 2 was not stimulated by polyinosic-polycytidylic acid in the PKR-K/R cells. RANKL stimulated the formation of TRAP-positive multinuclear cells in RAW264.7 cells. However, TRAP-positive multinuclear cells were not formed in the PKR-K/R cells even when the cells were stimulated with higher doses of RANKL. A specific inhibitor of PKR, 2-aminopurine, also suppressed the RANKL-induced osteoclast differentiation in RAW264.7 cells. The expression of macrophage fusion receptor and dendritic cell-specific transmembrane protein significantly decreased in the PKR-K/R cells by real time PCR analysis. The results of RT-PCR revealed that the mRNA expression of osteoclast markers (cathepsin K and calcitonin receptor) was suppressed in the PKR-K/R cells and RAW264.7 cells treated with 2-aminopurine. Expression of NF-{kappa}B protein was suppressed in the PKR-K/R cells and 2-aminopurine-treated RAW264.7 cells. The level of STAT1 protein expression was elevated in the PKR-K/R cells compared with that of the wild-type cells. Immunohistochemical study showed that PKR was localized in osteoclasts of metatarsal bone of newborn mouse. The finding that the PKR-positive multinuclear cells should be osteoclasts was confirmed by TRAP-staining. Our present study indicates that PKR plays important

  14. C60 fullerene localization and membrane interactions in RAW 264.7 immortalized mouse macrophages

    Science.gov (United States)

    Russ, K. A.; Elvati, P.; Parsonage, T. L.; Dews, A.; Jarvis, J. A.; Ray, M.; Schneider, B.; Smith, P. J. S.; Williamson, P. T. F.; Violi, A.; Philbert, M. A.

    2016-02-01

    There continues to be a significant increase in the number and complexity of hydrophobic nanomaterials that are engineered for a variety of commercial purposes making human exposure a significant health concern. This study uses a combination of biophysical, biochemical and computational methods to probe potential mechanisms for uptake of C60 nanoparticles into various compartments of living immune cells. Cultures of RAW 264.7 immortalized murine macrophage were used as a canonical model of immune-competent cells that are likely to provide the first line of defense following inhalation. Modes of entry studied were endocytosis/pinocytosis and passive permeation of cellular membranes. The evidence suggests marginal uptake of C60 clusters is achieved through endocytosis/pinocytosis, and that passive diffusion into membranes provides a significant source of biologically-available nanomaterial. Computational modeling of both a single molecule and a small cluster of fullerenes predicts that low concentrations of fullerenes enter the membrane individually and produce limited perturbation; however, at higher concentrations the clusters in the membrane causes deformation of the membrane. These findings are bolstered by nuclear magnetic resonance (NMR) of model membranes that reveal deformation of the cell membrane upon exposure to high concentrations of fullerenes. The atomistic and NMR models fail to explain escape of the particle out of biological membranes, but are limited to idealized systems that do not completely recapitulate the complexity of cell membranes. The surprising contribution of passive modes of cellular entry provides new avenues for toxicological research that go beyond the pharmacological inhibition of bulk transport systems such as pinocytosis.There continues to be a significant increase in the number and complexity of hydrophobic nanomaterials that are engineered for a variety of commercial purposes making human exposure a significant health concern

  15. Early transcriptional responses of internalization defective Brucella abortus mutants in professional phagocytes, RAW 264.7.

    Science.gov (United States)

    Cha, Seung Bin; Lee, Won Jung; Shin, Min Kyoung; Jung, Myung Hwan; Shin, Seung Won; Yoo, An Na; Kim, Jong Wan; Yoo, Han Sang

    2013-06-27

    Brucella abortus is an intracellular zoonotic pathogen which causes undulant fever, endocarditis, arthritis and osteomyelitis in human and abortion and infertility in cattle. This bacterium is able to invade and replicate in host macrophage instead of getting removed by this defense mechanism. Therefore, understanding the interaction between virulence of the bacteria and the host cell is important to control brucellosis. Previously, we generated internalization defective mutants and analyzed the envelope proteins. The present study was undertaken to evaluate the changes in early transcriptional responses between wild type and internalization defective mutants infected mouse macrophage, RAW 264.7. Both of the wild type and mutant infected macrophages showed increased expression levels in proinflammatory cytokines, chemokines, apoptosis and G-protein coupled receptors (Gpr84, Gpr109a and Adora2b) while the genes related with small GTPase which mediate intracellular trafficking was decreased. Moreover, cytohesin 1 interacting protein (Cytip) and genes related to ubiquitination (Arrdc3 and Fbxo21) were down-regulated, suggesting the survival strategy of this bacterium. However, we could not detect any significant changes in the mutant infected groups compared to the wild type infected group. In summary, it was very difficult to clarify the alterations in host cellular transcription in response to infection with internalization defective mutants. However, we found several novel gene changes related to the GPCR system, ubiquitin-proteosome system, and growth arrest and DNA damages in response to B. abortus infection. These findings may contribute to a better understanding of the molecular mechanisms underlying host-pathogen interactions and need to be studied further.

  16. The impact of transmission errors on progressive 720 lines HDTV coded with H.264

    Science.gov (United States)

    Brunnström, Kjell; Stålenbring, Daniel; Pettersson, Martin; Gustafsson, Jörgen

    2010-02-01

    TV sent over the networks based on the Internet Protocol i.e IPTV is moving towards high definition (HDTV). There has been quite a lot of work on how the HDTV is affected by different codecs and bitrates, but the impact of transmission errors over IP-networks have been less studied. The study was focusing on H.264 encoded 1280x720 progressive HDTV format and was comparing three different concealment methods for different packet loss rates. One is included in a propriety decoder, one is part of FFMPEG and different length of freezing. The target is to simulate what typically IPTV settop-boxes will do when encountering packet loss. Another aim is to study whether the presentation upscaled on the full HDTV screen or presented pixel mapped in a smaller area in the center of the sceen would have an effect on the quality. The results show that there were differences between the two packet loss concealment methods in FFMPEG and in the propriety codec. Freezing seemed to have similar effect as been reported before. For low rates of transmission errors the coding impairments has impact on the quality, but for higher degree of transmission errors these does not affect the quality, since they become overshadowed by transmission error. An interesting effect where the higher bitrate videos goes from having higher quality for lower degree of packet loss, to having lower quality than the lower bitrate video at higher packet loss, was discovered. The different way of presenting the video i.e. upscaled or not-upscaled was significant on the 95% level, but just about.

  17. Dynamic changes in the spatiotemporal localization of Rab21 in live RAW264 cells during macropinocytosis.

    Directory of Open Access Journals (Sweden)

    Youhei Egami

    2009-08-01

    Full Text Available Rab21, a member of the Rab GTPase family, is known to be involved in membrane trafficking, but its implication in macropinocytosis is unclear. We analyzed the spatiotemporal localization of Rab21 in M-CSF-stimulated RAW264 macrophages by the live-cell imaging of fluorescent protein-fused Rab21. It was demonstrated that wild-type Rab21 was transiently associated with macropinosomes. Rab21 was recruited to the macropinosomes after a decrease in PI(4,5P(2 and PI(3,4,5P(3 levels. Although Rab21 was largely colocalized with Rab5, the recruitment of Rab21 to the macropinosomes lagged a minute behind that of Rab5, and preceded that of Rab7. Then, Rab21 was dissociated from the macropinosomes prior to the accumulation of Lamp1, a late endosomal/lysosomal marker. Our analysis of Rab21 mutants revealed that the GTP-bound mutant, Rab21-Q78L, was recruited to the macropinosomes, similarly to wild-type Rab21. However, the GDP-bound mutant, Rab21-T33N, did not localize on the formed macropinosomes, suggesting that the binding of GTP to Rab21 is required for the proper recruitment of Rab21 onto the macropinosomes. However, neither mutation of Rab21 significantly affected the rate of macropinosome formation. These data indicate that Rab21 is a transient component of early and intermediate stages of macropinocytosis, and probably functions in macropinosome maturation before fusing with lysosomal compartments.

  18. Oxidized phospholipids induce ceramide accumulation in RAW 264.7 macrophages: role of ceramide synthases.

    Directory of Open Access Journals (Sweden)

    Lingaraju M Halasiddappa

    Full Text Available Oxidized phospholipids (OxPLs, including 1-palmitoyl-2-glutaroyl-sn-glycero-3-phosphocholine (PGPC and 1-palmitoyl-2-oxovaleroyl-sn-glycero-3-phosphocholine (POVPC are among several biologically active derivatives that are generated during oxidation of low-density lipoproteins (LDLs. These OxPLs are factors contributing to pro-atherogenic effects of oxidized LDLs (OxLDLs, including inflammation, proliferation and death of vascular cells. OxLDL also elicits formation of the lipid messenger ceramide (Cer which plays a pivotal role in apoptotic signaling pathways. Here we report that both PGPC and POVPC are cytotoxic to cultured macrophages and induce apoptosis in these cells which is associated with increased cellular ceramide levels after several hours. In addition, exposure of RAW 264.7 cells to POVPC and PGPC under the same conditions resulted in a significant increase in ceramide synthase activity, whereas, acid or neutral sphingomyelinase activities were not affected. PGPC is not only more toxic than POVPC, but also a more potent inducer of ceramide formation by activating a limited subset of CerS isoforms. The stimulated CerS activities are in line with the C16-, C22-, and C24:0-Cer species that are generated under the influence of the OxPL. Fumonisin B1, a specific inhibitor of CerS, suppressed OxPL-induced ceramide generation, demonstrating that OxPL-induced CerS activity in macrophages is responsible for the accumulation of ceramide. OxLDL elicits the same cellular ceramide and CerS effects. Thus, it is concluded that PGPC and POVPC are active components that contribute to the capacity of this lipoprotein to elevate ceramide levels in macrophages.

  19. Effects of bisphosphonates on osteoclastogenesis in RAW264.7 cells.

    Science.gov (United States)

    Abe, Keigo; Yoshimura, Yoshitaka; Deyama, Yoshiaki; Kikuiri, Takashi; Hasegawa, Tomokazu; Tei, Kanchu; Shinoda, Hisashi; Suzuki, Kuniaki; Kitagawa, Yoshimasa

    2012-06-01

    Bisphosphonates are used as therapeutic agents for the management of osteoporosis and other bone diseases. However, the precise effects and mechanisms of bisphosphonates on osteoclastogenesis are unclear, as previous studies have reported contradictory findings and no studies have circumstantially assessed the effects of bisphosphonates on osteoclastogenesis. Therefore, the aim of this study was to determine the effects of bisphosphonates on osteoclastogenesis in RAW264.7 (RAW) cells. To examine the direct effects of bisphosphonates on osteoclast differentiation via receptor activator of nuclear factor-κB (RANK) ligand (RANKL), RAW cells were cultured with bisphosphonates. Addition of bisphosphonates to RAW cells led to a significant decrease in the number of osteoclasts and large osteoclasts (≥ 8 nuclei) in a bisphosphonate concentration-dependent and time-dependent manner. The cytotoxicity of non-nitrogen-containing bisphosphonates was specific to osteoclasts, while nitrogen-containing bisphosphonates were cytotoxic and induced cell death in both osteoclasts and RAW cells. Resorption activity was significantly diminished by treatment with bisphosphonates, thus confirming that bisphosphonates impair the absorptive activity of osteoclasts. We also investigated the effects of bisphosphonates on the mRNA expression of genes associated with osteoclastogenesis, osteoclast-specific markers and apoptosis-related genes using quantitative real-time PCR. The results suggest that bisphosphonates suppress osteoclast differentiation and infusion, and induce osteoclast apoptosis. With regard to osteoclast apoptosis induced by bisphosphonates, we further investigated the detection of DNA fragmentation and Caspase-Glo 3/7 assay. DNA fragmentation was confirmed after treatment with bisphosphonates, while caspase-3/7 activity increased significantly when compared with controls. In conclusion, bisphosphonates directly inhibited RANKL-stimulated osteoclast differentiation and

  20. Quality optimization of H.264/AVC video transmission over noisy environments using a sparse regression framework

    Science.gov (United States)

    Pandremmenou, K.; Tziortziotis, N.; Paluri, S.; Zhang, W.; Blekas, K.; Kondi, L. P.; Kumar, S.

    2015-03-01

    We propose the use of the Least Absolute Shrinkage and Selection Operator (LASSO) regression method in order to predict the Cumulative Mean Squared Error (CMSE), incurred by the loss of individual slices in video transmission. We extract a number of quality-relevant features from the H.264/AVC video sequences, which are given as input to the LASSO. This method has the benefit of not only keeping a subset of the features that have the strongest effects towards video quality, but also produces accurate CMSE predictions. Particularly, we study the LASSO regression through two different architectures; the Global LASSO (G.LASSO) and Local LASSO (L.LASSO). In G.LASSO, a single regression model is trained for all slice types together, while in L.LASSO, motivated by the fact that the values for some features are closely dependent on the considered slice type, each slice type has its own regression model, in an e ort to improve LASSO's prediction capability. Based on the predicted CMSE values, we group the video slices into four priority classes. Additionally, we consider a video transmission scenario over a noisy channel, where Unequal Error Protection (UEP) is applied to all prioritized slices. The provided results demonstrate the efficiency of LASSO in estimating CMSE with high accuracy, using only a few features. les that typically contain high-entropy data, producing a footprint that is far less conspicuous than existing methods. The system uses a local web server to provide a le system, user interface and applications through an web architecture.

  1. Adapting hierarchical bidirectional inter prediction on a GPU-based platform for 2D and 3D H.264 video coding

    Science.gov (United States)

    Rodríguez-Sánchez, Rafael; Martínez, José Luis; Cock, Jan De; Fernández-Escribano, Gerardo; Pieters, Bart; Sánchez, José L.; Claver, José M.; de Walle, Rik Van

    2013-12-01

    The H.264/AVC video coding standard introduces some improved tools in order to increase compression efficiency. Moreover, the multi-view extension of H.264/AVC, called H.264/MVC, adopts many of them. Among the new features, variable block-size motion estimation is one which contributes to high coding efficiency. Furthermore, it defines a different prediction structure that includes hierarchical bidirectional pictures, outperforming traditional Group of Pictures patterns in both scenarios: single-view and multi-view. However, these video coding techniques have high computational complexity. Several techniques have been proposed in the literature over the last few years which are aimed at accelerating the inter prediction process, but there are no works focusing on bidirectional prediction or hierarchical prediction. In this article, with the emergence of many-core processors or accelerators, a step forward is taken towards an implementation of an H.264/AVC and H.264/MVC inter prediction algorithm on a graphics processing unit. The results show a negligible rate distortion drop with a time reduction of up to 98% for the complete H.264/AVC encoder.

  2. Activation of AMPA receptor promotes TNF-α release via the ROS-cSrc-NFκB signaling cascade in RAW264.7 macrophages

    Energy Technology Data Exchange (ETDEWEB)

    Cheng, Xiu-Li [Department of Physiology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences, School of Basic Medicine, Peking Union Medical College, Beijing (China); Ding, Fan [Office of Scientific R& D, Tsinghua University, Beijing (China); Li, Hui; Tan, Xiao-Qiu [Department of Physiology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences, School of Basic Medicine, Peking Union Medical College, Beijing (China); Liu, Xiao [Department of Pathophysiology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences, School of Basic Medicine, Peking Union Medical College, Beijing (China); Cao, Ji-Min, E-mail: caojimin@126.com [Department of Physiology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences, School of Basic Medicine, Peking Union Medical College, Beijing (China); Gao, Xue, E-mail: longlongnose@163.com [Department of Pathophysiology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences, School of Basic Medicine, Peking Union Medical College, Beijing (China)

    2015-05-29

    The relationship between glutamate signaling and inflammation has not been well defined. This study aimed to investigate the role of AMPA receptor (AMPAR) in the expression and release of tumor necrosis factor-alpha (TNF-α) from macrophages and the underlying mechanisms. A series of approaches, including confocal microscopy, immunofluorescency, flow cytometry, ELISA and Western blotting, were used to estimate the expression of AMPAR and downstream signaling molecules, TNF-α release and reactive oxygen species (ROS) generation in the macrophage-like RAW264.7 cells. The results demonstrated that AMPAR was expressed in RAW264.7 cells. AMPA significantly enhanced TNF-α release from RAW264.7 cells, and this effect was abolished by CNQX (AMPAR antagonist). AMPA also induced elevation of ROS production, phosphorylation of c-Src and activation of nuclear factor (NF)-κB in RAW264.7 cells. Blocking c-Src by PP2, scavenging ROS by glutathione (GSH) or inhibiting NF-κB activation by pyrrolidine dithiocarbamate (PDTC) decreased TNF-α production from RAW264.7 cells. We concluded that AMPA promotes TNF-α release in RAW264.7 macrophages likely through the following signaling cascade: AMPAR activation → ROS generation → c-Src phosphorylation → NF-κB activation → TNF-α elevation. The study suggests that AMPAR may participate in macrophage activation and inflammation. - Highlights: • AMPAR is expressed in RAW264.7 macrophages and is upregulated by AMPA stimulation. • Activation of AMPAR stimulates TNF-α release in macrophages through the ROS-cSrc-NFκB signaling cascade. • Macrophage AMPAR signaling may play an important role in inflammation.

  3. Chlorogenic acid protects against atherosclerosis in ApoE-/- mice and promotes cholesterol efflux from RAW264.7 macrophages.

    Directory of Open Access Journals (Sweden)

    Chongming Wu

    Full Text Available Chlorogenic acid (CGA is one of the most abundant polyphenols in the human diet and is suggested to be a potential antiatherosclerotic agent due to its proposed hypolipidemic, anti-inflammatory and antioxidative properties. The aim of this study was to evaluate the effect of CGA on atherosclerosis development in ApoE(-/- mice and its potential mechanism. ApoE(-/- mice were fed a cholesterol-rich diet without (control or with CGA (200 and 400 mg/kg or atorvastatin (4 mg/kg for 12 weeks. During the study plasma lipid and inflammatory parameters were determined. Treatment with CGA (400 mg/kg reduced atherosclerotic lesion area and vascular dilatation in the aortic root, comparable to atorvastatin. CGA (400 mg/kg also significantly decreased plasma levels of total cholesterol, triglycerides and low-density lipoprotein-cholesterol as well as inflammatory markers. Supplementation with CGA or CGA metabolites-containing serum suppressed oxidized low-density lipoprotein (oxLDL-induced lipid accumulation and stimulated cholesterol efflux from RAW264.7 cells. CGA significantly increased the mRNA levels of PPARγ, LXRα, ABCA1 and ABCG1 as well as the transcriptional activity of PPARγ. Cholesterol efflux assay showed that three major metabolites, caffeic, ferulic and gallic acids, significantly stimulated cholesterol efflux from RAW264.7 cells. These results suggest that CGA potently reduces atherosclerosis development in ApoE(-/- mice and promotes cholesterol efflux from RAW264.7 macrophages. Caffeic, ferulic and gallic acids may be the potential active compounds accounting for the in vivo effect of CGA.

  4. Modulation of inducible nitric oxide synthase gene expression in RAW 264.7 murine macrophages by Pacific ciguatoxin.

    Science.gov (United States)

    Kumar-Roiné, Shilpa; Matsui, Mariko; Chinain, Mireille; Laurent, Dominique; Pauillac, Serge

    2008-08-01

    To investigate the possible involvement of the nitric oxide radical (NO) in ciguatera fish poisoning (CFP), the in vitro effects of the main Pacific ciguatoxin (P-CTX-1B) and bacterial lipopolysaccharide (LPS) were comparatively studied on neuroblastoma Neuro-2a and on macrophage RAW 264.7 cell lines. NO accumulation was quantified by measuring nitrite levels in cellular supernatant using Griess reagent while the up-regulation of inducible nitric oxide synthase (iNOS) at the mRNA level was quantified via Real-Time Reverse-Transcription Polymerase Chain Reaction (RT-PCR). P-CTX-1B caused a concentration- and time-dependent induction of iNOS in RAW 264.7 cells but not in Neuro-2a cells. NO production was evidenced by increased nitrite levels in the 10 microM range after 48 h of RAW 264.7 cells exposure to LPS and P-CTX-1B (0.05 microg/ml and 6 nM, respectively). The expression of iNOS mRNA peaked at 8h for LPS then gradually decreased to low level at 48 h. In contrast, a sustained level was recorded with P-CTX-1B in the 8-48 h time interval. The addition of N(omega)-nitro-L-arginine methyl ester (L-NAME), a stereoselective NOS inhibitor, strongly diminished NO formation but had no effect on iNOS mRNA synthesis. The implication of NO in CFP paves the way for new therapies for both western and traditional medicines.

  5. Mechanisms of suppression of inducible nitric oxide synthase (iNOS) expression in RAW 264.7 cells by andrographolide

    Science.gov (United States)

    Chiou, Wen-Fei; Chen, Chieh-Fu; Lin, Jin-Jung

    2000-01-01

    Andrographolide, an active component found in leaves of Andrographis paniculata, has been reported to exhibit nitric oxide (NO) inhibitory property in endotoxin-stimulated macrophages, however, the detailed mechanisms remain unclear. In the present study we investigated the effect of andrographolide on the expression of inducible NO synthase (iNOS) mRNA, protein, and enzyme activity in RAW 264.7 macrophages stimulated with lipopolysaccharide (LPS) plus interferon-γ (IFN-γ).RAW 264.7 cells stimulated with LPS/IFN-γ activated NO production; in this condition andrographolide (1–100 μM) inhibited NO production in a dose-dependent manner with an IC50 value of 17.4±1.1 μM. Andrographolide also reduces the expression of iNOS protein level but without a significant effect on iNOS mRNA. The reduction of iNOS activity is thought to be caused by decreased expression of iNOS protein.In a protein stability assay, andrographolide moderately but significantly reduced the amount of iNOS protein as suggested by accelerating degradation. Furthermore, andrographolide also inhibited total protein de novo synthesis as demonstrated by [35S]-methionine incorporation.As a whole, these data suggest that andrographolide inhibits NO synthesis in RAW 264.7 cells by reducing the expression of iNOS protein and the reduction could occur through two additional mechanisms: prevention of the de novo protein synthesis and decreasing the protein stability via a post-transcriptional mechanism. It is also possible that inhibition of iNOS protein expression and NO production under immune stimulation and/or bacteria infection may explain, in part, the beneficial effects of andrographolide as an anti-inflammatory agent. PMID:10780958

  6. Anti-inflammatory effects of linalool in RAW 264.7 macrophages and lipopolysaccharide-induced lung injury model.

    Science.gov (United States)

    Huo, Meixia; Cui, Xiurui; Xue, Jiangdong; Chi, Gefu; Gao, Ruijie; Deng, Xuming; Guan, Shuang; Wei, Jingyuan; Soromou, Lanan Wassy; Feng, Haihua; Wang, Dacheng

    2013-03-01

    Inflammation, characterized by redness, swelling, pain and a sensation of heat, is one of the body's self-defense systems. Although the inflammation response has an important role in host survival, it also leads to chronic inflammatory diseases. Linalool is a natural compound of the essential oils in several aromatic plants species. It possesses anti-inflammatory, antinociceptive, and other bioactive properties. In the present study, we investigated the protective effects of linalool on inflammation in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells and an LPS-induced in vivo lung injury model. We evaluated the effects of linalool on LPS-induced production of inflammatory mediators in Raw 264.7 murine macrophages by enzyme-linked immunosorbent assay and Western blot. To confirm the anti-inflammatory activity of linalool in vivo, we induced an acute lung injury in an LPS-induced mouse model. Linalool attenuated the production of LPS-induced tumor necrosis-α and interleukin-6 both in vitro and in vivo. Furthermore, phosphorylation of IκBα protein, p38, c-Jun terminal kinase, and extracellular signal-regulated kinase in LPS-stimulated RAW 264.7 cells was blocked by linalool. Our in vivo study also found that linalool attenuated lung histopathologic changes in mouse models. The results suggest that linalool inhibits inflammation both in vitro and in vivo, and may be a potential therapeutic candidate for the treatment of inflammatory diseases. Copyright © 2013 Elsevier Inc. All rights reserved.

  7. Anti-inflammatory potential of peat moss extracts in lipopolysaccharide-stimulated RAW 264.7 macrophages.

    Science.gov (United States)

    Choi, Woo-Suk; Jeong, Jin-Woo; Kim, Sung Ok; Kim, Gi-Young; Kim, Byung-Woo; Kim, Cheol Min; Seo, Yong-Bae; Kim, Woe-Yeon; Lee, Sang-Yeol; Jo, Kwon-Ho; Choi, Young Ju; Choi, Yung Hyun; Kim, Gun-Do

    2014-10-01

    The aim of the present study was to identify the anti-inflammatory and anti-oxidative effects of peat moss aqueous extract (PME) on lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. To demonstrate the anti-inflammatory and antioxidant effects of PME, the levels of nitric oxide (NO) and cytokines were measured using Griess reagent and cytokine ELISA kits, respectively. Reverse transcriptase-polymerase chain reaction (RT-PCR) and western blot analysis were conducted to evaluate the expression of genes and proteins. Immunofluorescence was used to measure the expression and translocation of transcription factors. Pre-treatment with PME inhibited the production of prostaglandin E(2) and NO by suppressing the gene expression of cyclooxygenase-2 and inducible NO synthase, respectively. The LPS-stimulated gene expression and the production of tumor necrosis factor-α and interleukin-1β were significantly reduced by PME. In the LPS-stimulated RAW 264.7 cells, nuclear factor‑κB (NF-κB) translocated from the cytosol to the nucleus, while pre-treatment with PME induced the sequestration of NF-κB in the cytosol through the inhibition of IκBα degradation. In the same manner, PME contributed to the inhibition of the activation of mitogen-activated protein kinases. In addition, the PME-treated RAW 264.7 cells facilitated the activation of nuclear factor-like 2 (Nrf2) , and in turn, enhanced heme oxygenase-1 (HO-1) expression. These results indicate that PME exerts anti-inflammatory and antioxidant effects, and suggest that PME may neutralize inflammation and prevent cellular damage by oxidative stress.

  8. Efficient bragg grating fabrication in Ge-rich fibre by high-intensity femtosecond 264 mm irradation

    DEFF Research Database (Denmark)

    Slattery, S.A.; Nikogasyan, D.N.; Plougmann, Nikolai

    2004-01-01

    Fibre Bragg gratings (FBGs) are usually fabricated by UV laser light (typically, 248 nm radiation from a KrF* excimer laser), which spectrally coincides with the absorption band of defects in germanosilicate fibre core [1, 2]. Recently, it was demonstrated that the use of highintensity femtosecond...... silica glass and corresponding defects [5]. In this Letter we study FBG recording in a highly-nonlinear fibre (HNLF, OFS Denmark), employing 264 nm femtosecond pulses with a peak intensity of around 180 GW=cm2. refractive fluence, kJ/cm2....

  9. Pyrrolizidine alkaloids from Liparis nervosa with inhibitory activities against LPS-induced NO production in RAW264.7 macrophages.

    Science.gov (United States)

    Huang, Shuai; Zhou, Xian-li; Wang, Cui-juan; Wang, You-song; Xiao, Feng; Shan, Lian-hai; Guo, Zhi-yun; Weng, Jie

    2013-09-01

    Six pyrrolizidine alkaloids were isolated from the whole herb of Liparis nervosa together with two previously known ones. Their structures were elucidated by extensive spectroscopic analyses and chemical reactions. The cytotoxicity of the isolates was evaluated against A549, HepG2, and MCF-7 human cancer cell lines; however, no significant growth inhibition was observed. All compounds were evaluated for the inhibition of LPS-induced nitric oxide (NO) production in RAW264.7 macrophages, and most significantly inhibited NO production with IC50 values in the range of 2.16-38.25 μM. Copyright © 2013 Elsevier Ltd. All rights reserved.

  10. A model of R-D performance evaluation for Rate-Distortion-Complexity evaluation of H.264 video coding

    DEFF Research Database (Denmark)

    Wu, Mo; Forchhammer, Søren

    2007-01-01

    This paper considers a method for evaluation of Rate-Distortion-Complexity (R-D-C) performance of video coding. A statistical model of the transformed coefficients is used to estimate the Rate-Distortion (R-D) performance. A model frame work for rate, distortion and slope of the R-D curve for inter...... and intra frame is presented. Assumptions are given for analyzing an R-D model for fast R-D-C evaluation. The theoretical expressions are combined with H.264 video coding, and confirmed by experimental results. The complexity frame work is applied to the integer motion estimation....

  11. Phenolic amides from Tribulus terrestris and their inhibitory effects on nitric oxide production in RAW 264.7 cells.

    Science.gov (United States)

    Kim, Hyung Sik; Lee, Jin Woo; Jang, Hari; Le, Thi Phuong Linh; Kim, Jun Gu; Lee, Moon Soon; Hong, Jin Tae; Lee, Mi Kyeong; Hwang, Bang Yeon

    2018-02-01

    A new phenolic amide, named cis-terrestriamide (7), together with ten known compounds (1-6, 8-11), were isolated from the methanolic extract of the fruits of Tribulus terrestris. The structure of 7 was elucidated on the basis of extensive analyses of 1D and 2D nuclear magnetic resonance spectroscopic and high resolution mass spectrometry data. Compounds 1, 2, 5, 6, 8, 9, and 11 exhibited inhibitory effects on the lipopolysaccharide-stimulated nitric oxide production in RAW 264.7 cells, with IC 50 values of 18.7-49.4 μM.

  12. On Optimizing H. 264/AVC Rate Control by Improving R-D Model and Incorporating HVS Characteristics

    Directory of Open Access Journals (Sweden)

    Jiang Gangyi

    2010-01-01

    Full Text Available The state-of-the-art JVT-G012 rate control algorithm of H.264 is improved from two aspects. First, the quadratic rate-distortion (R-D model is modified based on both empirical observations and theoretical analysis. Second, based on the existing physiological and psychological research findings of human vision, the rate control algorithm is optimized by incorporating the main characteristics of the human visual system (HVS such as contrast sensitivity, multichannel theory, and masking effect. Experiments are conducted, and experimental results show that the improved algorithm can simultaneously enhance the overall subjective visual quality and improve the rate control precision effectively.

  13. Association of the I264T Variant in the Sulfide Quinone Reductase-Like (SQRDL) Gene with Osteoporosis in Korean Postmenopausal Women

    Science.gov (United States)

    Park, Eunkuk; Kim, Bo-Young; Choi, Vit-Na; Yoo, Young-Hyun; Kim, Bom-Taeck; Jeong, Seon-Yong

    2015-01-01

    To identify novel susceptibility variants for osteoporosis in Korean postmenopausal women, we performed a genome-wide association analysis of 1180 nonsynonymous single nucleotide polymorphisms (nsSNPs) in 405 individuals with osteoporosis and 722 normal controls of the Korean Association Resource cohort. A logistic regression analysis revealed 72 nsSNPs that showed a significant association with osteoporosis (posteoporosis. To assess whether the SQRDL I264T variant played a functional role in the pathogenesis of osteoporosis, we examined the in vitro effect of the nsSNP on bone remodeling. Overexpression of the SQRDL I264T variant in the preosteoblast MC3T3-E1 cells significantly increased alkaline phosphatase activity, mineralization, and the mRNA expression of osteoblastogenesis markers, Runx2, Sp7, and Bglap genes, whereas the SQRDL wild type had no effect or a negative effect on osteoblast differentiation. Overexpression of the SQRDL I264T variant did not affect osteoclast differentiation of the primary-cultured monocytes. The known effects of hydrogen sulfide (H2S) on bone remodeling may explain the findings of the current study, which demonstrated the functional role of the H2S-catalyzing enzyme SQRDL I264T variant in osteoblast differentiation. In conclusion, the results of the statistical and experimental analyses indicate that the SQRDL I264T nsSNP may be a significant susceptibility variant for osteoporosis in Korean postmenopausal women that is involved in osteoblast differentiation. PMID:26258864

  14. A Prototyping Virtual Socket System-On-Platform Architecture with a Novel ACQPPS Motion Estimator for H.264 Video Encoding Applications

    Directory of Open Access Journals (Sweden)

    Qiu Yifeng

    2009-01-01

    Full Text Available H.264 delivers the streaming video in high quality for various applications. The coding tools involved in H.264, however, make its video codec implementation very complicated, raising the need for algorithm optimization, and hardware acceleration. In this paper, a novel adaptive crossed quarter polar pattern search (ACQPPS algorithm is proposed to realize an enhanced inter prediction for H.264. Moreover, an efficient prototyping system-on-platform architecture is also presented, which can be utilized for a realization of H.264 baseline profile encoder with the support of integrated ACQPPS motion estimator and related video IP accelerators. The implementation results show that ACQPPS motion estimator can achieve very high estimated image quality comparable to that from the full search method, in terms of peak signal-to-noise ratio (PSNR, while keeping the complexity at an extremely low level. With the integrated IP accelerators and optimized techniques, the proposed system-on-platform architecture sufficiently supports the H.264 real-time encoding with the low cost.

  15. Association of the I264T variant in the sulfide quinone reductase-like (SQRDL) gene with osteoporosis in Korean postmenopausal women.

    Science.gov (United States)

    Jin, Hyun-Seok; Kim, Jeonghyun; Park, Sangwook; Park, Eunkuk; Kim, Bo-Young; Choi, Vit-Na; Yoo, Young-Hyun; Kim, Bom-Taeck; Jeong, Seon-Yong

    2015-01-01

    To identify novel susceptibility variants for osteoporosis in Korean postmenopausal women, we performed a genome-wide association analysis of 1180 nonsynonymous single nucleotide polymorphisms (nsSNPs) in 405 individuals with osteoporosis and 722 normal controls of the Korean Association Resource cohort. A logistic regression analysis revealed 72 nsSNPs that showed a significant association with osteoporosis (posteoporosis. To assess whether the SQRDL I264T variant played a functional role in the pathogenesis of osteoporosis, we examined the in vitro effect of the nsSNP on bone remodeling. Overexpression of the SQRDL I264T variant in the preosteoblast MC3T3-E1 cells significantly increased alkaline phosphatase activity, mineralization, and the mRNA expression of osteoblastogenesis markers, Runx2, Sp7, and Bglap genes, whereas the SQRDL wild type had no effect or a negative effect on osteoblast differentiation. Overexpression of the SQRDL I264T variant did not affect osteoclast differentiation of the primary-cultured monocytes. The known effects of hydrogen sulfide (H2S) on bone remodeling may explain the findings of the current study, which demonstrated the functional role of the H2S-catalyzing enzyme SQRDL I264T variant in osteoblast differentiation. In conclusion, the results of the statistical and experimental analyses indicate that the SQRDL I264T nsSNP may be a significant susceptibility variant for osteoporosis in Korean postmenopausal women that is involved in osteoblast differentiation.

  16. Comparative assessment of H.265/MPEG-HEVC, VP9, and H.264/MPEG-AVC encoders for low-delay video applications

    Science.gov (United States)

    Grois, Dan; Marpe, Detlev; Nguyen, Tung; Hadar, Ofer

    2014-09-01

    The popularity of low-delay video applications dramatically increased over the last years due to a rising demand for realtime video content (such as video conferencing or video surveillance), and also due to the increasing availability of relatively inexpensive heterogeneous devices (such as smartphones and tablets). To this end, this work presents a comparative assessment of the two latest video coding standards: H.265/MPEG-HEVC (High-Efficiency Video Coding), H.264/MPEG-AVC (Advanced Video Coding), and also of the VP9 proprietary video coding scheme. For evaluating H.264/MPEG-AVC, an open-source x264 encoder was selected, which has a multi-pass encoding mode, similarly to VP9. According to experimental results, which were obtained by using similar low-delay configurations for all three examined representative encoders, it was observed that H.265/MPEG-HEVC provides significant average bit-rate savings of 32.5%, and 40.8%, relative to VP9 and x264 for the 1-pass encoding, and average bit-rate savings of 32.6%, and 42.2% for the 2-pass encoding, respectively. On the other hand, compared to the x264 encoder, typical low-delay encoding times of the VP9 encoder, are about 2,000 times higher for the 1-pass encoding, and are about 400 times higher for the 2-pass encoding.

  17. Real-time video streaming using H.264 scalable video coding (SVC) in multihomed mobile networks: a testbed approach

    Science.gov (United States)

    Nightingale, James; Wang, Qi; Grecos, Christos

    2011-03-01

    Users of the next generation wireless paradigm known as multihomed mobile networks expect satisfactory quality of service (QoS) when accessing streamed multimedia content. The recent H.264 Scalable Video Coding (SVC) extension to the Advanced Video Coding standard (AVC), offers the facility to adapt real-time video streams in response to the dynamic conditions of multiple network paths encountered in multihomed wireless mobile networks. Nevertheless, preexisting streaming algorithms were mainly proposed for AVC delivery over multipath wired networks and were evaluated by software simulation. This paper introduces a practical, hardware-based testbed upon which we implement and evaluate real-time H.264 SVC streaming algorithms in a realistic multihomed wireless mobile networks environment. We propose an optimised streaming algorithm with multi-fold technical contributions. Firstly, we extended the AVC packet prioritisation schemes to reflect the three-dimensional granularity of SVC. Secondly, we designed a mechanism for evaluating the effects of different streamer 'read ahead window' sizes on real-time performance. Thirdly, we took account of the previously unconsidered path switching and mobile networks tunnelling overheads encountered in real-world deployments. Finally, we implemented a path condition monitoring and reporting scheme to facilitate the intelligent path switching. The proposed system has been experimentally shown to offer a significant improvement in PSNR of the received stream compared with representative existing algorithms.

  18. Anti-Inflammatory Activity of the Methanol Extract of Moutan Cortex in LPS-Activated Raw264.7 Cells

    Directory of Open Access Journals (Sweden)

    Seung-Chul Chun

    2007-01-01

    Full Text Available Moutan Cortex (MCE has been used in traditional medicine to remove heat from the blood, promote blood circulation and alleviate blood stasis. This study was conducted to evaluate the effects of MCE on regulatory mechanisms of cytokines and nitric oxide (NO involved in immunological activity of Raw264.7 cells. Cells were pretreated with methanolic extracts of MCE, and further cultured for an appropriate time after lipopolyssacharide (LPS addition. During the entire experimental period, 0.1 and 0.3 mg ml−1 of MCE had no cytotoxicity. In these concentrations, MCE inhibited the production of NO and prostaglandin E2 (PGE2, the expression of inducible NO synthase (iNOS, cyclooxygenase-2 (COX-2 and phosphorylated inhibitor of κBα (p-IκBα, and the activation of nuclear factor κB (NF-κB. MCE also reduced the concentration of tumor necrosis factor-α (TNF-α, interleukin-1β (IL-1β and interleukin-6 (IL-6 in the Raw264.7 cells that were activated by LPS. These results demonstrate that MCE has anti-inflammatory effects through the inhibition of iNOS and COX-2 expression by suppressing the phosphorylation of I-κBα and the activation of NF-κB.

  19. Sulforaphane suppresses LPS-induced or TPA-induced downregulation of PDCD4 in RAW 264.7 cells.

    Science.gov (United States)

    Cho, Jong-Ho; Kim, Young-Woo; Keum, Young-Sam

    2014-11-01

    Sulforaphane is a natural chemopreventive isothiocyanate and abundantly found in various cruciferous vegetables. Although chemopreventive activity of sulforaphane is well documented, the detailed biochemical mechanism(s), underlying how it regulates the protein translation process to antagonize pro-inflammatory responses are largely unclear. In the present study, we show that lipopolysaccharide (LPS) or 12-O-tetradecanoylphorbol-13-acetate (TPA) treatment reduces cellular levels of PDCD4, and this event is mediated by affecting both transcription and proteolysis in RAW 264.7 cells. We show that LPS-mediated or TPA-mediated PDCD4 downregulation is catalyzed by the activation of intracellular Akt1 or S6K1 kinases and that sulforaphane suppresses LPS-induced or TPA-induced Akt1 or S6K1 activation, thereby resulting in the attenuation of PDCD4 downregulation in RAW 264.7 cells. We propose that sulforaphane suppression of PDCD4 downregulation serves as a novel molecular mechanism to control proliferation in response to pro-inflammatory signals. Copyright © 2014 John Wiley & Sons, Ltd.

  20. The Effects of Bee Venom on PLA2 and Calcium Concentration in Raw 264.7 Cells

    Directory of Open Access Journals (Sweden)

    Jong-Il Yun

    2003-06-01

    Full Text Available Objectives : The purpose of this study was to investigate the effect of Bee Venom on the lipopolysaccharide, sodium nitroprusside and hydrogen peroxide induced expression phospholipase A2 and calcium concentration in RAW 264.7 cells, a murine macrophage cell line. Methods : The expression of phospholipase A2 was determined by western blotting with corresponding antibodies, and the generation of intracellular calcium concentration was investigated by delta scan system in RAW 264.7 cells. Results : 1. Compared with control, expressions of lipopolysaccharide-induced phospholipase A2 were decreased significantly by 1 ㎍/㎕ of bee venom and decreased by 0.5, 5 ㎍/㎕ of bee venom. 2. Compared with control, expressions of sodium nitroprusside-induced phospholipase A2 were decreased significantly by 5 ㎍/㎕ of bee venom but increased by 0.5, 5 ㎍/㎕ of bee venom. 3. Compared with control, expressions of hydrogen peroxide-induced phospholipase A2 were decreased significaltly by 1 ㎍/㎕ of bee venom and decreased by 0.5 ㎍/㎕ of bee venom but increased by 5 ㎍/㎕ of bee venom. 4. Compared with control, lipopolysaccharide, sodium nitroprusside and hydrogen peroxide- induced intracellular calcium concentrations were decreased by 0.5, 1, 5 ㎍/㎕ of bee venom and by indomethacin

  1. Algorithm and Hardware Design of a Fast Intra Frame Mode Decision Module for H.264/AVC Encoders

    Directory of Open Access Journals (Sweden)

    Daniel Palomino

    2012-01-01

    Full Text Available In the rate-distortion optimization (RDO, the process of choosing the best prediction mode is performed through exhaustive executions of the whole encoding process, increasing significantly the encoder computational complexity. Considering H.264/AVC intra frame prediction, there are several modes to encode a macroblock (MB. This work proposes an algorithm and the hardware design for a fast intra frame mode decision module for H.264/AVC encoders. The application of the proposed algorithm reduces in more than 10 times the number of encoding iterations for choosing the best intramode when compared with RDO-based decision. The architecture was synthesized to FPGA and achieved an operation frequency of 98 MHz processing more than 300 HD1080p frames per second. With this approach, we achieved one order-of-magnitude performance improvement compared with RDO-based approaches, which is very important not only from the performance but also from the energy consumption perspective for battery-operated devices. In order to compare the architecture with previously published works, we also synthesized it to standard cells. Compared with the best previous results reported, the implemented architecture achieves a complexity reduction of five times, a processing capability increase of 14 times, and a reduction in the number of clock cycles per MB of 11 times.

  2. Anti‐Inflammatory Effect of Quercetin on RAW 264.7 Mouse Macrophages Induced with Polyinosinic‐Polycytidylic Acid

    Directory of Open Access Journals (Sweden)

    Young‐Jin Kim

    2016-04-01

    Full Text Available Quercetin (3,3′,4′,5,6‐pentahydroxyflavone is a well‐known antioxidant and a flavonol found in many fruits, leaves, and vegetables. Quercetin also has known anti‐inflammatory effects on lipopolysaccharide‐induced macrophages. However, the effects of quercetin on virus‐induced macrophages have not been fully reported. In this study, the anti‐inflammatory effect of quercetin on double‐stranded RNA (dsRNA‐induced macrophages was examined. Quercetin at concentrations up to 50 μM significantly inhibited the production of NO, IL‐6, MCP‐1, IP‐10, RANTES, GM‐CSF, G‐CSF, TNF‐α, LIF, LIX, and VEGF as well as calcium release in dsRNA (50 μg/mL of polyinosinic‐polycytidylic acid‐induced RAW 264.7 mouse macrophages (p < 0.05. Quercetin at concentrations up to 50 μM also significantly inhibited mRNA expression of signal transducer and activated transcription 1 (STAT1 and STAT3 in dsRNA‐induced RAW 264.7 cells (p < 0.05. In conclusion, quercetin had alleviating effects on viral inflammation based on inhibition of NO, cytokines, chemokines, and growth factors in dsRNA‐induced macrophages via the calcium‐STAT pathway.

  3. Effects of CCR5-Delta32, CCR2-64I, and SDF-1 3'A alleles on HIV-1 disease progression

    DEFF Research Database (Denmark)

    Ioannidis, J P; Rosenberg, P S; Goedert, J J

    2001-01-01

    seroconversion. Data were combined with fixed-effects and random-effects models. RESULTS: Both the CCR5-Delta32 and CCR2-64I alleles were associated with a decreased risk for progression to AIDS (relative hazard among seroconverters, 0.74 and 0.76, respectively; P = 0.01 for both), a decreased risk for death...... (relative hazard among seroconverters, 0.64 and 0.74; P CCR5-Delta32 or CCR2-64I allele had no clear protective effect on the risk for death.......00), or death after development of AIDS (relative hazard, 0.81 and 0.97; P > 0.5 for all). CONCLUSIONS: The CCR5-Delta32 and CCR2-64I alleles had a strong protective effect on progression of HIV-1 infection, but SDF-1 3'A homozygosity carried no such protection....

  4. Suppression of the lipopolysaccharide-induced expression of MARCKS-related protein (MRP) affects transmigration in activated RAW264.7 cells.

    Science.gov (United States)

    Chun, Kwang-Rok; Bae, Eun Mi; Kim, Jae-Kwan; Suk, Kyoungho; Lee, Won-Ha

    2009-01-01

    The molecular action mechanism of MRP, one of the protein kinase C (PKC) substrates, has been under intense investigation, but reports on its role in macrophage function remain controversial. The treatment of macrophage cell lines with bacterial lipopolysaccharide (LPS) induced a high level of MRP expression suggesting that MRP plays a role in the function of activated macrophages. In order to investigate the role of MRP in activated RAW264.7 cells, we stably transfected MRP-specific shRNA expression constructs and tested for alterations in macrophage-related functions. The down-regulation of MRP expression resulted in a marked reduction in chemotaxis toward MCP-1 or extracellular matrix proteins. Furthermore, pharmacological inhibitors of PKC significantly inhibited the chemotaxis in RAW264.7 cells. These data reveals the pivotal role of MRP in the transmigration of activated RAW264.7 cells.

  5. Enantioselective Total Synthesis of Antibiotic CJ-16,264, Synthesis and Biological Evaluation of Designed Analogues, and Discovery of Highly Potent and Simpler Antibacterial Agents.

    Science.gov (United States)

    Nicolaou, K C; Pulukuri, Kiran Kumar; Rigol, Stephan; Buchman, Marek; Shah, Akshay A; Cen, Nicholas; McCurry, Megan D; Beabout, Kathryn; Shamoo, Yousif

    2017-11-08

    An improved and enantioselective total synthesis of antibiotic CJ-16,264 through a practical kinetic resolution and an iodolactonization reaction to form the iodo pyrrolizidinone fragment of the molecule is described. A series of racemic and enantiopure analogues of CJ-16,264 was designed and synthesized through the developed synthetic technologies and tested against drug-resistant bacterial strains. These studies led to interesting structure-activity relationships and the identification of a number of simpler, and yet equipotent, or even more potent, antibacterial agents than the natural product, thereby setting the foundation for further investigations in the quest for new anti-infective drugs.

  6. [Blood pressure in 6- to 45-month-old children. Apropos of a study of 264 children from nurseries].

    Science.gov (United States)

    Bérard, E; Boutté, P; Macone, F; Albertini, M; Mariani, R

    1988-03-01

    The systolic, diastolic and mean blood pressure and pulse rate were measured among 264 children aged 6 to 45 months, during routine check-up in 4 day-nurseries. We took the mean value of 3 successive measurements taken at one minute intervals on the upper arm of calm, asymptomatic children, with an automatic oscillometric monitor (Dinamap). When the mean systolic blood pressure was greater than 110 mmHg, the measurement was repeated. The results were related to weight, height and age and to personal and/or family history. The systolic pressure was steady with a 97th percentile at 110 mmHg. The diastolic pressure presented two levels: 97th percentile at 81 mmHg before 24 months and 73 mmHg afterwards. Nine children born to toxemic mothers, and 14 having required neonatal hospital care, had lower diastolic pressure. The other family or personal data were not linked to particular pressure groups.

  7. Global, regional, and national age-sex specific mortality for 264 causes of death, 1980–2016

    DEFF Research Database (Denmark)

    Moesgaard Iburg, Kim

    2017-01-01

    (CODEm) to generate estimates for each location, year, age, and sex. We assessed observed and expected levels and trends of cause-specific deaths in relation to the Socio-demographic Index (SDI), a summary indicator derived from measures of average income per capita, educational attainment, and total......Background Monitoring levels and trends in premature mortality is crucial to understanding how societies can address prominent sources of early death. The Global Burden of Disease 2016 Study (GBD 2016) provides a comprehensive assessment of cause-specific mortality for 264 causes in 195 locations...... from 1980 to 2016. This assessment includes evaluation of the expected epidemiological transition with changes in development and where local patterns deviate from these trends. Methods We estimated cause-specific deaths and years of life lost (YLLs) by age, sex, geography, and year. YLLs were...

  8. Micrometam C Protects against Oxidative Stress in Inflammation Models in Zebrafish and RAW264.7 Macrophages

    Directory of Open Access Journals (Sweden)

    Hao Tang

    2015-08-01

    Full Text Available Micrometam C is a core of novel marine compound isolated from the mangrove associates Micromelum falcatum. In this study, we investigated the protective effects of micrometam C in inflammation models in the transgenic zebrafish line Tg (corola: eGFP and RAW264.7 macrophages. We found that micrometam C significantly suppressed the migration of immune cells in tail-cutting-induced inflammation in transgenic zebrafish and reduced lipopolysaccharide (LPS-induced reactive oxygen species (ROS in both zebrafish and macrophages. In addition, micrometam C also restored LPS-induced reduction of endogenous antioxidants, such as catalase (CAT, glutathione (GSH and superoxide dismutase (SOD. The protective effects of micrometam C were in parallel to its inhibition of NADPH oxidase and nuclear factor-kappa-binding (NF-κB activity. Thus, the present results demonstrate that micrometam C protects against LPS-induced inflammation possibly through its antioxidant property.

  9. [Different expression of the Asn264LysfsX35 mutation of the GNAS gene in a family with pseudohypoparathyroidism.].

    Science.gov (United States)

    de Arriba Muñoz, A; Labarta Aizpún, J I; Dehesa, E Mayayo; Calvo, M T; Pérez de Nanclares, G; Ferrández-Longás, A

    2011-02-01

    Pseudohypoparathyroidism (PHP) is a heterogeneous group of endocrine diseases characterised by hypocalcaemia, hyperphosphataemia and resistance to PTH. There are different forms of PHP. PHP-Ia is the most frequent form and shows multi-hormonal resistance, GNAS (Gs(α)) mutations and signs of Albright́s hereditary osteodystrophy (AHO). PseudoPHP (PPHP) have isolated AHO without hormonal resistance and it is also caused by GNAS mutations. We present a family that share the same inactivating GNAS mutation (Asn264LysfsX35); the mother being affected with PPHP and the two daughters with PHP-Ia. We discuss the different clinical phenotypes and the dominant mode of inheritance with genetic imprinting where the phenotype of the offspring depends on the sex of the parent affected. Copyright © 2010 Asociación Española de Pediatría. Published by Elsevier Espana. All rights reserved.

  10. New Sesquiterpene Oxidations with CYP260A1 and CYP264B1 from Sorangium cellulosum So ce56.

    Science.gov (United States)

    Schifrin, Alexander; Litzenburger, Martin; Ringle, Michael; Ly, Thuy T B; Bernhardt, Rita

    2015-12-01

    Sesquiterpenes are natural products derived from the common precursor farnesyl pyrophosphate (FPP) but are highly diverse in structure and function. Cytochrome P450 enzymes (P450s) exhibit the unique ability to introduce molecular oxygen into non-activated C-H bonds. In plant biosynthetic pathways, P450s commonly derivatize sesquiterpene hydrocarbons. However, the potential of bacterial P450s for terpene derivatization is still underinvestigated. This work compares the substrate specificities and regioselectivities of the sesquiterpene hydroxylases CYP260A1 and CYP264B1 from myxobacterium Sorangium cellulosum So ce56. Four tested substrate classes (eremophilanes, humulanes, caryophyllanes, and cedranes) were converted by both P450s. The achievable variety of oxidations is demonstrated on the model substrates (+)-nootkatone and zerumbone. Increasing the number of functionally investigated P450s, this study represents a step towards the selective derivatization of sesquiterpenes. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  11. Fast mode decision based on human noticeable luminance difference and rate distortion cost for H.264/AVC

    Science.gov (United States)

    Li, Mian-Shiuan; Chen, Mei-Juan; Tai, Kuang-Han; Sue, Kuen-Liang

    2013-12-01

    This article proposes a fast mode decision algorithm based on the correlation of the just-noticeable-difference (JND) and the rate distortion cost (RD cost) to reduce the computational complexity of H.264/AVC. First, the relationship between the average RD cost and the number of JND pixels is established by Gaussian distributions. Thus, the RD cost of the Inter 16 × 16 mode is compared with the predicted thresholds from these models for fast mode selection. In addition, we use the image content, the residual data, and JND visual model for horizontal/vertical detection, and then utilize the result to predict the partition in a macroblock. From the experimental results, a greater time saving can be achieved while the proposed algorithm also maintains performance and quality effectively.

  12. Effects of florfenicol on LPS-induced nitric oxide and prostaglandin E₂ production in RAW 264.7 macrophages.

    Science.gov (United States)

    Zhang, Xuemei; Xiong, Huanzhang; Li, Hongyu; Yu, Lu; Deng, Xuming

    2011-10-01

    Florfenicol, an antibiotic commonly used to treat infections, has previously been shown to modulate lipopolysaccharide (LPS)-induced early cytokine responses by blocking the nuclear factor-κB (NF-κB) pathway. In this study, we investigated the effects of florfenicol on nitric oxide (NO) and prostaglandin E₂ (PGE₂) production as well as on inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) expression in LPS-stimulated murine RAW 264.7 macrophages. We also analysed the effects of florfenicol on mitogen-activated protein kinase (MAPK) pathways. Florfenicol significantly inhibited LPS-induced NO and PGE₂ production. Consistent with these observations, mRNA and protein expression of iNOS and COX-2 were also inhibited by florfenicol in a dose-dependent manner. Furthermore, phosphorylation of p38 and extracellular signal-regulated kinase 1/2 (ERK1/2) in LPS-stimulated RAW 264.7 cells was suppressed by florfenicol. However, c-Jun N-terminal kinase (JNK) phosphorylation remained unaffected. Using specific inhibitors of ERK and p38, we found that florfenicol may inhibit NO and PGE₂ mostly through ERK and p38 pathway. These results suggest that florfenicol inhibits NO and PGE₂ production in conjunction with an inhibition of iNOS and COX-2 expression, at least partially via suppression of ERK1/2 and p38 MAPK phosphorylation. © 2010 The Authors Fundamental and Clinical Pharmacology © 2010 Société Française de Pharmacologie et de Thérapeutique.

  13. Riboflavin deprivation inhibits macrophage viability and activity - a study on the RAW 264.7 cell line.

    Science.gov (United States)

    Mazur-Bialy, Agnieszka Irena; Buchala, Beata; Plytycz, Barbara

    2013-08-28

    Riboflavin, or vitamin B2, as a precursor of the coenzymes FAD and FMN, has an indirect influence on many metabolic processes and determines the proper functioning of several systems, including the immune system. In the human population, plasma riboflavin concentration varies from 3·1 nM (in a moderate deficiency, e.g. in pregnant women) to 10·4 nM (in healthy adults) and 300 nM (in cases of riboflavin supplementation). The purpose of the present study was to investigate the effects of riboflavin concentration on the activity and viability of macrophages, i.e. on one of the immunocompetent cell populations. The study was performed on the murine monocyte/macrophage RAW 264.7 cell line cultured in medium with various riboflavin concentrations (3·1, 10·4, 300 and 531 nM). The results show that riboflavin deprivation has negative effects on both the activity and viability of macrophages and reduces their ability to generate an immune response. Signs of riboflavin deficiency developed in RAW 264.7 cells within 4 d of culture in the medium with a low riboflavin concentration (3·1 nM). In particular, the low riboflavin content reduced the proliferation rate and enhanced apoptotic cell death connected with the release of lactate dehydrogenase. The riboflavin deprivation impaired cell adhesion, completely inhibited the respiratory burst and slightly impaired phagocytosis of the zymosan particles. In conclusion, macrophages are sensitive to riboflavin deficiency; thus, a low riboflavin intake in the diet may affect the immune system and may consequently decrease proper host immune defence.

  14. Proteomic Investigation of the Time Course Responses of RAW 264.7 Macrophages to Infection with Salmonella enterica

    Energy Technology Data Exchange (ETDEWEB)

    Shi, Liang; Chowdhury, Saiful M.; Smallwood, Heather S.; Yoon, Hyunjin; Mottaz-Brewer, Heather M.; Norbeck, Angela D.; McDermott, Jason E.; Clauss, Therese RW; Heffron, Fred; Smith, Richard D.; Adkins, Joshua N.

    2009-08-01

    Macrophages plan important roles in controlling Salmonella-mediated systemic infection. To investigate the responses of macrophages to Salmonella infection, we infected RAW 264.7 macrophages with Salmonella enterica serovar Typhimurium (STM) and then performed a comparative liquid chromatography-tandem mass spectrometry [LC-MS(/MS)]-based proteomics analysis of the infected macrophages. A total of 1006 macrophage and 115 STM proteins were indentified from this study. Most of STM proteins were found at late stage of the time course of infection, consistent with the fact that STM proliferates inside RAW 264.7 macrophages. Majority of the identified macrophage proteins were house keeping-related, including cytoplasmic superoxide dismutase 1 (SOD1), whose peptide abundances were relatively constant during the time course of infection. Compared to those in no infection control, the peptide abundances of 244 macrophage proteins (or 24% of total indentified macrophage proteins) changed considerably after STM infection. The functions of these STM infection-affected macrophage proteins were diverse and ranged from production of antibacterial nitric oxide (i.e., inducible nitric oxide synthase or iNOS) or production of prostaglandin H2 (i.e., prostaglandin-endoperoxide synthase 2, also know as cyclooxygenase-2 or COX-2) to regulation of intracellular traffic (e.g., sorting nexin or SNX 5, 6 and 9), demonstrating a global impact of STM infection on macrophage proteome. Western-blot analysis not only confirmed the LC-MS(/MS) results of SOD1, COX-2 and iNOS, but also revealed that the protein abundances of mitochondrial SOD2 increased after STM infection, indicating an infection-induced oxidative stress in mitochondria.

  15. Curcumin suppresses the production of interleukin-6 in Prevotella intermedia lipopolysaccharide-activated RAW 264.7 cells.

    Science.gov (United States)

    Kim, Sung-Jo

    2011-06-01

    Curcumin is known to exert numerous biological effects including anti-inflammatory activity. In this study, we investigated the effects of curcumin on the production of interleukin-6 (IL-6) by murine macrophage-like RAW 264.7 cells stimulated with lipopolysaccharide (LPS) from Prevotella intermedia, a major cause of inflammatory periodontal disease, and sought to determine the underlying mechanisms of action. LPS was prepared from lyophilized P. intermedia ATCC 25611 cells by the standard hot phenol-water method. Culture supernatants were collected and assayed for IL-6. We used real-time polymerase chain reaction to detect IL-6 mRNA expression. IκB-α degradation, nuclear translocation of NF-κB subunits, and STAT1 phosphorylation were characterized via immunoblotting. DNA-binding of NF-κB was also analyzed. Curcumin strongly suppressed the production of IL-6 at both gene transcription and translation levels in P. intermedia LPS-activated RAW 264.7 cells. Curcumin did not inhibit the degradation of IκB-α induced by P. intermedia LPS. Curcumin blocked NF-κB signaling through the inhibition of nuclear translocation of NF-κB p50 subunit. Curcumin also attenuated DNA binding activity of p50 and p65 subunits and suppressed STAT1 phosphorylation. Although further study is required to explore the detailed mechanism of action, curcumin may contribute to blockade of the host-destructive processes mediated by IL-6 and appears to have potential therapeutic values in the treatment of inflammatory periodontal disease.

  16. Microarray-based identification of differentially expressed genes in intracellular Brucella abortus within RAW264.7 cells.

    Directory of Open Access Journals (Sweden)

    Mingxing Tian

    Full Text Available Brucella spp. is a species of facultative intracellular Gram-negative bacteria that induces abortion and causes sterility in domesticated mammals and chronic undulant fever in humans. Important determinants of Brucella's virulence and potential for chronic infection include the ability to circumvent the host cell's internal surveillance system and the capability to proliferate within dedicated and non-dedicated phagocytes. Hence, identifying genes necessary for intracellular survival may hold the key to understanding Brucella infection. In the present study, microarray analysis reveals that 7.82% (244/3334 of all Brucella abortus genes were up-regulated and 5.4% (180/3334 were down-regulated in RAW264.7 cells, compared to free-living cells in TSB. qRT-PCR verification further confirmed a >5-fold up-regulation for fourteen genes. Functional analysis classified araC, ddp, and eryD as to partake in information storage and processing, alp, flgF and virB9 to be involved in cellular processes, hpcd and aldh to play a role in metabolism, mfs and nikC to be involved in both cellular processes and metabolism, and four hypothetical genes (bruAb1_1814, bruAb1_0475, bruAb1_1926, and bruAb1_0292 had unknown functions. Furthermore, we constructed a B. abortus 2308 mutant Δddp where the ddp gene is deleted in order to evaluate the role of ddp in intracellular survival. Infection assay indicated significantly higher adherence and invasion abilities of the Δddp mutant, however it does not survive well in RAW264.7 cells. Brucella may survive in hostile intracellular environment by modulating gene expression.

  17. Comparison of LTQ-Orbitrap Velos and Q-Exactive for proteomic analysis of 1-1000 ng RAW 264.7 cell lysate digests

    Science.gov (United States)

    Sun, Liangliang; Zhu, Guijie

    2013-01-01

    Rational There is interest in extending bottom-up proteomics to the smallest possible sample size. We investigated the performance of two modern instruments for the analysis of samples ranging from 1 ng to 1 µg of RAW 264.7 cell lysate digests. Methods A UPLC system coupled with either an LTQ-Orbitrap Velos or a Q-Exactive mass spectrometers was used for peptide separation and identification. Results For 1–1,000 ng RAW 264.7 cell lysate digests, the Q-Exactive generated 10–83% more protein groups and 11–109% more peptides than the LTQ-Orbitrap Velos (higher-energy collisional dissociation, HCD) with MASCOT database searching, due to faster scan rate and higher resolution. In addition, HCD and collision induced dissociation (CID) modes of the LTQ-Orbitrap Velos were compared. HCD produced higher peptide and protein group IDs than CID for 1–1,000 ng RAW 264.7 cell lysate digests with MASCOT database searching. Database searching results from SEQUEST and MASCOT were also compared and comparable protein group IDs were obtained from the two search engines. Conclusions These results indicated that Q-Exactive outperformed LTQ-Orbitrap Velos for shotgun proteomics analysis of 1 to 1,000 ng RAW 264.7 cell lysate digests in terms of obtained peptide and protein group IDs. PMID:23239329

  18. Reactive oxygen species are the major antibacterials against Salmonella Typhimurium purine auxotrophs in the phagosome of RAW 264.7 cells.

    Science.gov (United States)

    Mantena, Radha K R; Wijburg, Odilia L C; Vindurampulle, Christofer; Bennett-Wood, Vicki R; Walduck, Anna; Drummond, Grant R; Davies, John K; Robins-Browne, Roy M; Strugnell, Richard A

    2008-05-01

    Intramacrophage survival appears to be a pathogenic trait common to Salmonellae and definition of the metabolic requirements of Salmonella within macrophages might provide opportunities for novel therapeutic interventions. We show that loss of PurG function in Salmonella enterica serovar Typhimurium SL1344 leads to death of the bacterium in RAW264.7 cells, which was due to unavailability of purine nucleotides but not thiamine in the phagosome of RAW264.7 cells. Phagosomal escape of purG mutant restored growth, suggesting that the phagosomal environment, but not the cytosol, is toxic to Salmonella purine auxotrophs. NADPH oxidase inhibition restored the growth of purG mutant in RAW264.7 cells, implying that the Salmonella-containing vacuole acquires reactive oxygen species (ROS) that are lethal to purine auxotrophs. Under purine limiting conditions, purG mutant was unable to repair the damage caused by hydrogen peroxide or UV irradiation, suggesting that ROS-mediated DNA damage may have been responsible for the attenuated phenotype of purG mutant in RAW264.7 cells and in mice. These studies highlight the possibility of utilizing the Salmonella purine nucleotide biosynthetic pathway as a prospective therapeutic target and also underline the importance of metabolic pathways in assembling a comprehensive understanding of the host-pathogen interactions inside phagocytic cells.

  19. Human p53(264-272) HLA-A2 binding peptide is an immunodominant epitope in DNA-immunized HLA-A2 transgenic mice

    DEFF Research Database (Denmark)

    Petersen, T R; Bregenholta, S; Pedersen, L O

    1999-01-01

    C57BL/10 mice transgenic for HLA-A2 were immunized with either a full-length DNA-construct of the tumor suppressor p53 or with a minigene encoding the p53-derived immunodominant peptide p53(264)LLGRNSFEV272 (L9V). Vaccination with the full-length p53 construct induced potent cytotoxic activity...

  20. Effects of CCR5-Delta32 and CCR2-64I alleles on HIV-1 disease progression: the protection varies with duration of infection

    NARCIS (Netherlands)

    Mulherin, Stephanie A.; O'Brien, Thomas R.; Ioannidis, John P.; Goedert, James J.; Buchbinder, Susan P.; Coutinho, Roel A.; Jamieson, Beth D.; Meyer, Laurence; Michael, Nelson L.; Pantaleo, Giuseppe; Rizzardi, G. Paolo; Schuitemaker, Hanneke; Sheppard, Haynes W.; Theodorou, Ioannis D.; Vlahov, David; Rosenberg, Philip S.

    2003-01-01

    OBJECTIVE: To examine temporal variation in the effects of CCR5-Delta32 and CCR2-64I chemokine receptor gene polymorphisms on HIV-1 disease progression. DESIGN: Pooled analysis of individual patient data from 10 cohorts of HIV-1 seroconverters from the United States, Europe, and Australia. METHODS:

  1. Causal pathways of the effects of age and the CCR5-Delta32, CCR2-64I, and SDF-1 3'A alleles on AIDS development

    NARCIS (Netherlands)

    Geskus, Ronald B.; Meyer, Laurence; Hubert, Jean-Baptiste; Schuitemaker, Hanneke; Berkhout, Ben; Rouzioux, Christine; Theodorou, Ioannis D.; Delfraissy, Jean-François; Prins, Maria; Coutinho, Roel A.

    2005-01-01

    OBJECTIVE: To investigate the causal pathways by which age and the CCR5-Delta32, CCR2-64I, and SDF-1 3'A alleles influence progression to AIDS. DESIGN: Analysis of follow-up data from 2 cohort studies among homosexual men (n=400), having >10 years of follow-up. METHODS: The effects of the 4

  2. Global, regional, and national age-sex specific mortality for 264 causes of death, 1980-2016 : a systematic analysis for the Global Burden of Disease Study 2016

    NARCIS (Netherlands)

    Hoek, H. W.; van Boven, Job; Postma, Maarten

    2017-01-01

    BACKGROUND: Monitoring levels and trends in premature mortality is crucial to understanding how societies can address prominent sources of early death. The Global Burden of Disease 2016 Study (GBD 2016) provides a comprehensive assessment of cause-specific mortality for 264 causes in 195 locations

  3. Aspirin inhibits osteoclastogenesis by suppressing the activation of NF-κB and MAPKs in RANKL-induced RAW264.7 cells.

    Science.gov (United States)

    Zeng, Yan-Ping; Yang, Chao; Li, Yuan; Fan, Yong; Yang, Hong-Jun; Liu, Bin; Sang, Hong-Xun

    2016-09-01

    Aspirin is a commonly used medicine as an effective antipyretic, analgesic and anti-inflammatory drug. Previous studies have demonstrated its potential effects of anti-postmenopausal osteoporosis, while the molecular mechanisms remain unclear. The effects of aspirin on receptor‑activator of nuclear factor κB (NF‑κB) ligand (RANKL)‑induced osteoclasts were investigated in RAW264.7 cells in the current study. Using tartrate‑resistant acid phosphatase (TRAP) staining, it was observed that aspirin inhibited the differentiation of RANKL‑induced RAW264.7 cells. The mRNA expression of osteoclastic marker genes, including cathepsin K, TRAP, matrix metalloproteinase 9 and calcitonin receptor, were suppressed by aspirin as identified using reverse transcription‑quantitative polymerase chain reaction analysis. The immunofluorescence assay indicated that aspirin markedly inhibited NF‑κB p65 translocation to the nucleus in RANKL‑induced RAW264.7 cells. In addition, aspirin also suppressed the phosphorylation of mitogen‑activated protein kinases (MAPKs), observed by western blot analysis. Taken together, these data identified that aspirin inhibits osteoclastogenesis by suppressing the activation of NF‑κB and MAPKs in RANKL‑induced RAW264.7 cells, implying that aspirin may possess therapeutic potential for use in the prevention and treatment of osteoporosis.

  4. Flavonoids Identified from Korean Scutellaria baicalensis Georgi Inhibit Inflammatory Signaling by Suppressing Activation of NF-κB and MAPK in RAW 264.7 Cells

    Directory of Open Access Journals (Sweden)

    Gyeong-Eun Hong

    2013-01-01

    Full Text Available Scutellaria baicalensis Georgi has been used as traditional medicine for treating inflammatory diseases, hepatitis, tumors, and diarrhea in Asia. Hence, we investigated the anti-inflammatory effect and determined the molecular mechanism of action of flavonoids isolated from Korean S. baicalensis G. in lipopolysaccharide- (LPS- stimulated RAW 264.7 macrophages. A 3-(4,5-dimethylthiazol-2-yl-2,5-diphenyltetrazolium bromide assay was performed to examine cytotoxicity of the flavonoids at various concentrations of 10, 40, 70, and 100 µg/mL. No cytotoxicity was observed in RAW 264.7 cells at these concentrations. Furthermore, the flavonoids decreased production of inflammatory mediators such as inducible nitric oxide synthase, cyclooxygenase-2, interleukin-6, and tumor necrosis factor-alpha and inhibited phosphorylation of nuclear factor-kappa B (NF-κB and mitogen-activated protein kinases (MAPKs in LPS-induced RAW 264.7 cells. Moreover, to identify the differentially expressed proteins in RAW 264.7 cells of the control, LPS-treated, and flavonoid-treated groups, two-dimensional gel electrophoresis and mass spectrometry were conducted. The identified proteins were involved in the inflammatory response and included PRKA anchor protein and heat shock protein 70 kD. These findings suggest that the flavonoids isolated from S. baicalensis G. might have anti-inflammatory effects that regulate the expression of inflammatory mediators by inhibiting the NF-κB signaling pathway via the MAPK signaling pathway in RAW 264.7 cells.

  5. [Construction of transgenic RAW264.7 monoclonal cell line by dual-labeling of +8-green fluorescent protein and modified red fluorescent protein phosphatidylserine].

    Science.gov (United States)

    Ren, L; Wang, Y Q; Michael, Glogauer

    2016-09-01

    To establish the transgenic RAW264.7 monoclone cell line labelled by green fluorescent protein(GFP) and modified red fluorescent protein(mRFP), for investigating the mechanism of cell membrane fusion and cell biological behavior during osteoclastogenesis. A dual-labeling technique involving GFP and mRFP was applied to RAW264.7 cell line by pVSVG for in situ monitoring of membrane fusion during osteoclastogenesis. The live-cell imaging technology was adopted to consecutively observe the process of osteoclast formation induced by receptor activator of NF-κB ligand(RANKL). Furthermore, tartrate-resistant acid phosphatase staining(TRAP) and 4',6-diamidino-2-phenylindole(DAPI) staining were also used to identify the function and characteristics of RAW264.7 monoclonecell line transfected with phosphatidylserine(PS) and + 8 membrane charge. The normal morphology of RAW264.7 monoclonecell line transfected with + 8-GFP and PS-mRFP was preserved. The PS and (8+) biosensors co-expressed on the membrane of monocytes. No significant difference of fluorescence density was found. In osteoclasts, (8 + )probes disappeared, while PS expressed in both internal organelles and membrane of osteoclasts. Live-cell imaging observation showed that the multinuclear osteoclasts were generated among monocytes and apocytes. All fusion processes occurred under the condition of cell adherence. Successful construction of transgenic RAW264.7 monoclone cell line by GFP and mRFP tags provided a wide field of vision for further investigating the cytoskeleton and organelles of subcellular spatial dimension in osteoclastogenesis.

  6. Immunostimulatory Activity of Opuntia ficus-indica var. Saboten Cladodes Fermented by Lactobacillus plantarum and Bacillus subtilis in RAW 264.7 Macrophages.

    Science.gov (United States)

    Hwang, Joon-Ho; Lim, Sang-Bin

    2017-02-01

    To increase the functionality of Opuntia ficus-indica var. saboten cladodes, it was fermented by Lactobacillus plantarum and Bacillus subtilis. Eighty percent methanol extracts were investigated for their effects on nitric oxide (NO) production, cytokine secretion, nuclear factor-κB (NF-κB) activity, and mitogen-activated protein kinase (MAPK) phosphorylation in RAW 264.7 cells. Methanol extracts of L. plantarum culture medium (LPCME) and B. subtilis culture medium (BSCME) did not affect lipopolysaccharide (LPS)-induced NO production but, at 500 μg/mL, increased interferon (IFN)-γ-induced NO production by 55.2 and 66.5 μM, respectively, in RAW 264.7 cells. In RAW 264.7 cells not treated with LPS and IFN-γ, LPCME did not affect NO production, but BSCME increased NO production significantly in a dose-dependent manner. In addition, BSCME induced the expression of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) in RAW 264.7 cells in a dose-dependent manner. BSCME at 500 μg/mL increased TNF-α and IL-1β mRNA levels by 83.8% and 82.2%, respectively. BSCME increased NF-κB-dependent luciferase activity in a dose-dependent manner; 500 μg/mL BSCME increased activity 9.1-fold compared with the control. BSCME induced the phosphorylation of p38, c-JUN NH 2 -terminal protein kinase (JNK), and extracellular signal-regulated kinase (ERK) in a dose-dependent manner, but did not affect total ERK levels. In conclusion, BSCME exerted immunostimulatory effects, which were mediated by MAPK phosphorylation and NF-κB activation, resulting in increased TNF-α and IL-1β gene expression in RAW 264.7 macrophages. Therefore, BSCM shows promise for use as an immunostimulatory therapeutic.

  7. BCG Increased Membrane Expression of TRIM59 Through the TLR2/ TLR4/IRF5 Pathway in RAW264.7 Macrophages.

    Science.gov (United States)

    Jin, Zheng; Tian, Yuan; Yan, Dongmei; Li, Dong; Zhu, Xun

    2017-01-01

    In our previous study, we showed that Bacillus Calmette-Guerin (BCG)- activated macrophages have the ability to directly kill tumor cells. One of the main properties of these macrophages is the high expression of tripartite motif family protein 59 (TRIM59). This study was conducted to investigate the mechanism of BCG-induced TRIM59 expression on macrophages and to identify the subcellular localization of TRIM59. TRIM59 expression and TNF-α secretion were compared in RAW264.7 macrophage cells that were stimulated using BCG with or without Toll-like receptor 2/4 (TLR2/4)-neutralizing antibodies. Next, small interfering RNA (siRNA) was used to down-regulated interferon regulatory factor 5 (IRF5) gene expression in RAW264.7 cells. Transfected cells were stimulated with BCG, after which TRIM59 expression and TNF-α secretion were evaluated in cells pre-treated with siRNA or scramble control. After treatments, supernatants were co-cultured with MCA207, and cell viabilities were determined. Moreover, BCG-stimulated RAW264.7 cells were stained for TRIM59 and F4/80 expression. In this study, we showed that TRIM59 was expressed on the membrane of RAW264.7 cells. After blocking TLR2/4, treatment with BCG failed to induce the expression of TRIM59, IRF5, and TNF-α on RAW264.7 cells. In addition, down-regulation of IRF5 inhibited TRIM59 and TNF-α expression. Our study showed that TRIM59 is a membrane protein, and that BCG treatment upregulated TRIM59 expression on macrophages via TLR2/4 and IRF5 pathways. Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.

  8. Low-Complexity Hierarchical Mode Decision Algorithms Targeting VLSI Architecture Design for the H.264/AVC Video Encoder

    Directory of Open Access Journals (Sweden)

    Guilherme Corrêa

    2012-01-01

    Full Text Available In H.264/AVC, the encoding process can occur according to one of the 13 intraframe coding modes or according to one of the 8 available interframes block sizes, besides the SKIP mode. In the Joint Model reference software, the choice of the best mode is performed through exhaustive executions of the entire encoding process, which significantly increases the encoder's computational complexity and sometimes even forbids its use in real-time applications. Considering this context, this work proposes a set of heuristic algorithms targeting hardware architectures that lead to earlier selection of one encoding mode. The amount of repetitions of the encoding process is reduced by 47 times, at the cost of a relatively small cost in compression performance. When compared to other works, the fast hierarchical mode decision results are expressively more satisfactory in terms of computational complexity reduction, quality, and bit rate. The low-complexity mode decision architecture proposed is thus a very good option for real-time coding of high-resolution videos. The solution is especially interesting for embedded and mobile applications with support to multimedia systems, since it yields good compression rates and image quality with a very high reduction in the encoder complexity.

  9. Coriandrum sativum L. seed extract mitigates lipotoxicity in RAW 264.7 cells and prevents atherogenic changes in rats

    Science.gov (United States)

    Patel, Dipak; Desai, Swati; Gajaria, Tejal; Devkar, Ranjitsinh; Ramachandran, A.V.

    2013-01-01

    This study was designed to assess the efficacy of Coriandrum sativum L. (CS) in preventing in vitro low density lipoprotein (LDL) oxidation mediated macrophage modification. Further, an in vivo study was also conducted to confirm upon the efficacy of CS seed extract in alleviating pathophysiological alterations of high cholesterol diet induced atherosclerosis in rats. Copper mediated cell free oxidation of LDL accounted for elevated indices of malondialdehyde (MDA), lipid hydroperoxide (LHP)and protein carbonyl (PC) and a progressive increment in conjugate diene (CD) levels whereas, reverse set of changes were recorded in presence of CS extract. Cell mediated LDL oxidation (using RAW 264.7 cells) accounted for lowered MDA production and oxidized LDL (Ox-LDL) mediated cell death in presence of CS extract and the same was attributed to its potent antioxidant and free radical scavenging potentials. High cholesterol fed atherogenic rats showed elevated lipid indices, evidences of LDL oxidation, plaque formation in thoracic aorta. The same was further validated with immunostaining of cell adhesion molecules and hematoxylin and eosin (HXE) staining. However, co-supplementation of CS to atherogenic rats recorded significant lowering of the above mentioned parameters further strengthening the claim that CS extract is instrumental in preventing onset and progression of atherosclerosis. PMID:26417232

  10. Anti-inflammatory effects of Passiflora foetida L. in LPS-stimulated RAW264.7 macrophages.

    Science.gov (United States)

    Park, Ji-Won; Kwon, Ok-Kyoung; Ryu, Hyung Won; Paik, Jin-Hyub; Paryanto, Imam; Yuniato, Prasetyawan; Choi, Sangho; Oh, Sei-Ryang; Ahn, Kyung-Seop

    2018-03-12

    Passiflora foetida L. (Passifloraceae), a perennial climber in general, is used for treating many ailments in conventional medicine. In this study, the anti-inflammatory effect of methanolic extracts of P. foetida L. (PFME) and the involvement of nuclear factor-κB (NF-κB) signalling in the regulation of inflammation were investigated. PFME prevented the production of prostaglandin E2 (PGE2) and the expression of inducible cyclooxygenase-2 (COX-2) in lipopolysaccharide (LPS)-induced macrophage cells. Additionally, PFME reduced the release of pro-inflammatory cytokines. Moreover, in LPS-induced RAW264.7 cells, the phosphorylation of MAPKs (ERK1/2, p38 and JNK) was suppressed by PFME. Furthermore, PFME inhibited the NF-κB activation induced by LPS, which was associated with nuclear p65 levels with the abrogation of IκBα degradation and subsequent decreases. These results indicated that the PFME inhibited the LPS-induced inflammatory and oxidative responses. Therefore, we propose that the PFME may be therapeutic for treating inflammatory diseases.

  11. Structural characterization of blackberry wine polysaccharides and immunomodulatory effects on LPS-activated RAW 264.7 macrophages.

    Science.gov (United States)

    Cordeiro Caillot, Adriana Rute; de Lacerda Bezerra, Iglesias; Palhares, Laís Cristina Gusmão Ferreira; Santana-Filho, Arquimedes Paixão; Chavante, Suely Ferreira; Sassaki, Guilherme Lanzi

    2018-08-15

    Three polysaccharide fractions were isolated from blackberry wine. The crude extract BWPs was obtained with ethanol precipitation and freeze-thawing process, it was then submitted to Fehling treatment, giving soluble BWPFs and insoluble BWPFp fractions. These fractions were characterized by Gas Chromatography-Mass Spectrometry (GC-MS) and Nuclear Magnetic Resonance (NMR). Major polysaccharides were identified for each fraction: mannan, type II arabinogalactan and type I rhamnogalacturonan for BWPs, a mannan formed by a major chain of α-Manp(1 → 6)-linked units, O-2 substituted with α-d-Manp(1 → 2)-linked side chains for BWPFp and a AG II formed by a major chain of β-d-Galp(1 → 3)-linked, substituted at O-6 by side chains of the β-d-Galp(1 → 6)-linked, which then are substituted at O-3 by non-reducing units of α-l-Araf and a RG I, formed by [→4)-α-d-GalpA-(1 → 2)-α-l-Rhap-(1→] n for BWPFs. Anti-inflammatory effects of polysaccharide fractions were evaluated in RAW 264.7 cells. Fractions markedly reduced nitric oxide (NO) and pro-inflammatory cytokine production (TNF-α and IL-1β) in LPS-treated cells. Copyright © 2018 Elsevier Ltd. All rights reserved.

  12. Antioxidant and anti-inflammatory effects in RAW264.7 macrophages of malvidin, a major red wine polyphenol.

    Directory of Open Access Journals (Sweden)

    Eszter Bognar

    Full Text Available Red wine polyphenols can prevent cardiovascular and inflammatory diseases. Resveratrol, the most extensively studied constituent, is unlikely to solely account for these beneficial effects because of its rather low abundance and bioavailability. Malvidin is far the most abundant polyphenol in red wine; however, very limited data are available about its effect on inflammatory processes and kinase signaling pathways. METHODS FINDINGS: The present study was carried out by using RAW 264.7 macrophages stimulated by bacterial lipopolysaccharide in the presence and absence of malvidin. From the cells, activation of nuclear factor-kappaB, mitogen-activated protein kinase, protein kinase B/Akt and poly ADP-ribose polymerase, reactive oxygen species production, mitogen-activated protein kinase phosphatase-1 expression and mitochondrial depolarization were determined. We found that malvidin attenuated lipopolysaccharide-induced nuclear factor-kappaB, poly ADP-ribose polymerase and mitogen-activated protein kinase activation, reactive oxygen species production and mitochondrial depolarization, while upregulated the compensatory processes; mitogen-activated protein kinase phosphatase-1 expression and Akt activation.These effects of malvidin may explain the previous findings and at least partially account for the positive effects of moderate red wine consumption on inflammation-mediated chronic maladies such as obesity, diabetes, hypertension and cardiovascular disease.

  13. Cytoprotective and enhanced anti-inflammatory activities of liposomal piroxicam formulation in lipopolysaccharide-stimulated RAW 264.7 macrophages.

    Science.gov (United States)

    Chiong, Hoe Siong; Yong, Yoke Keong; Ahmad, Zuraini; Sulaiman, Mohd Roslan; Zakaria, Zainul Amiruddin; Yuen, Kah Hay; Hakim, Muhammad Nazrul

    2013-01-01

    Liposomal drug delivery systems, a promising lipid-based nanoparticle technology, have been known to play significant roles in improving the safety and efficacy of an encapsulated drug. Liposomes, prepared using an optimized proliposome method, were used in the present work to encapsulate piroxicam, a widely prescribed nonsteroidal anti-inflammatory drug. The cytotoxic effects as well as the in vitro efficacy in regulation of inflammatory responses by free-form piroxicam and liposome-encapsulated piroxicam were evaluated using a lipopolysaccharide-sensitive macrophage cell line, RAW 264.7. Cells treated with liposome-encapsulated piroxicam demonstrated higher cell viabilities than those treated with free-form piroxicam. In addition, the liposomal piroxicam formulation resulted in statistically stronger inhibition of pro-inflammatory mediators (ie, nitric oxide, tumor necrosis factor-α, interleukin-1β, and prostaglandin E2) than piroxicam at an equivalent dose. The liposome-encapsulated piroxicam also caused statistically significant production of interleukin-10, an anti-inflammatory cytokine. This study affirms the potential of a liposomal piroxicam formulation in reducing cytotoxicity and enhancing anti-inflammatory responses in vitro.

  14. Bentonite buffer pre-test. Core drilling of drillholes ONK-PP264...267 in ONKALO at Olkiluoto 2010

    International Nuclear Information System (INIS)

    Toropainen, V.

    2010-12-01

    Suomen Malmi Oy (Smoy) core drilled four drillholes for bentonite buffer pre-test in ONKALO at Eurajoki, Olkiluoto in July 2010. The identification numbers of the holes are ONK-PP264..267, and the lengths of the drillholes are approximately 4.30 metres each. The drillholes are 75.7 mm by diameter. The drillholes were drilled in a niche at access tunnel chainage 1475. The hydraulic DE 130 drilling rig was used for the work. The drilling water was taken from the ONKALO drilling water pipeline and premixed sodium fluorescein was used as a label agent in the drilling water. In addition to drilling, the drillcores were logged and reported by geologist. Geological logging included the following parameters: lithology, foliation, fracture parameters, fractured zones, core loss, weathering, fracture frequency, RQD and rock quality. The main rock type in the drillholes is pegmatitic granite. The average fracture frequency in the drill cores is 4.0 pcs / m and the average RQD value 94.2 %. (orig.)

  15. Iron Reduces M1 Macrophage Polarization in RAW264.7 Macrophages Associated with Inhibition of STAT1

    Directory of Open Access Journals (Sweden)

    Zhen-Shun Gan

    2017-01-01

    Full Text Available Iron metabolism in inflammation has been mostly characterized in macrophages exposed to pathogens or inflammatory conditions. The aim of this study is to investigate the cross-regulatory interactions between M1 macrophage polarization and iron metabolism. Firstly, we characterized the transcription of genes related to iron homeostasis in M1 RAW264.7 macrophages stimulated by IFN-γ. The molecular signature of M1 macrophages showed high levels of iron storage (ferritin, a low level of iron export (ferroportin, and changes of iron regulators (hepcidin and transferrin receptors, which favour iron sequestration in the reticuloendothelial system and are benefit for inflammatory disorders. Then, we evaluated the effect of iron on M1 macrophage polarization. Iron significantly reduced mRNA levels of IL-6, IL-1β, TNF-α, and iNOS produced by IFN-γ-polarized M1 macrophages. Immunofluorescence analysis showed that iron also reduced iNOS production. However, iron did not compromise but enhanced the ability of M1-polarized macrophages to phagocytose FITC-dextran. Moreover, we demonstrated that STAT1 inhibition was required for reduction of iNOS and M1-related cytokines production by the present of iron. Together, these findings indicated that iron decreased polarization of M1 macrophages and inhibited the production of the proinflammatory cytokines. The results expanded our knowledge about the role of iron in macrophage polarization.

  16. The Impact of Serum Amyloid P-Component on Gene Expression in RAW264.7 Mouse Macrophages

    Directory of Open Access Journals (Sweden)

    Dan Xi

    2016-01-01

    Full Text Available Serum amyloid P-component (SAP contributes to host defense and prevents fibrosis. Macrophages are the most abundant inflammatory cell type in atherosclerotic plaques. In the present study, using 3H-cholesterol-labeled counting radioactivity assay, we demonstrated that the apoAI-mediated cholesterol efflux in RAW264.7 macrophages was increased by SAP treatment in a time- and dose-dependent manner. We analyzed global gene expression changes upon SAP treatment using RNA sequencing. As a result, a total of 175 differentially expressed genes were identified, of which 134 genes were downregulated and 41 genes were upregulated in SAP treated cells compared to control cells. Quantitative RT-PCR analysis confirmed decreased expression of 5 genes and an increase in expression of 1 gene upon SAP treatment. Gene ontology analysis showed that genes involved in response to stimulus were significantly enriched in differentially expressed genes. Beyond protein-coding genes, we also identified 8 differentially expressed long noncoding RNAs. Our study may provide new insights into mechanisms underlying the functional role of SAP in macrophages.

  17. Echinacea Species and Alkamides Inhibit Prostaglandin E2 Production in RAW264.7 Mouse Macrophage Cells

    Science.gov (United States)

    LaLone, Carlie A.; Hammer, Kimberly D. P.; Wu, Lankun; Bae, Jaehoon; Leyva, Norma; Liu, Yi; Solco, Avery K. S.; Kraus, George A.; Murphy, Patricia A.; Wurtele, Eve S.; kim, Ok-Kyung; Seo, Kwon; Widrlechner, Mark P.; Birt, Diane F.

    2008-01-01

    Inhibition of prostaglandin E2 (PGE2) production in lipopolysaccharide-stimulated RAW264.7 mouse macrophage cells was assessed with an enzyme immunoassay following treatments with Echinacea extracts or synthesized alkamides. Results indicated that ethanol extracts diluted in media to a concentration of 15 μg/mL from E. angustifolia, E. pallida, E. simulata, and E. sanguinea significantly inhibited PGE2 production. In further studies, PGE2 production was significantly reduced by all synthesized alkamides assayed at 50 μM, by Bauer alkamides 8, 12A analogue, and 14, Chen alkamide 2, and Chen alkamide 2 analogue at 25 μM and by Bauer alkamide 14 at 10 μM. Cytotoxicity did not play a role in the noted reduction of PGE2 production in either the Echinacea extracts or synthesized alkamides. High-performance liquid chromatography analysis identified individual alkamides present at concentrations below 2.8 μM in the extracts from the six Echinacea species (15 μg/mL crude extract). Because active extracts contained Echinacea in a synergistic or additive manner. PMID:17696440

  18. Echinacea species and alkamides inhibit prostaglandin E(2) production in RAW264.7 mouse macrophage cells.

    Science.gov (United States)

    LaLone, Carlie A; Hammer, Kimberly D P; Wu, Lankun; Bae, Jaehoon; Leyva, Norma; Liu, Yi; Solco, Avery K S; Kraus, George A; Murphy, Patricia A; Wurtele, Eve S; Kim, Ok-Kyung; Seo, Kwon Ii; Widrlechner, Mark P; Birt, Diane F

    2007-09-05

    Inhibition of prostaglandin E(2) (PGE(2)) production in lipopolysaccharide-stimulated RAW264.7 mouse macrophage cells was assessed with an enzyme immunoassay following treatments with Echinacea extracts or synthesized alkamides. Results indicated that ethanol extracts diluted in media to a concentration of 15 microg/mL from E. angustifolia, E. pallida, E. simulata, and E. sanguinea significantly inhibited PGE2 production. In further studies, PGE2 production was significantly reduced by all synthesized alkamides assayed at 50 microM, by Bauer alkamides 8, 12A analogue, and 14, Chen alkamide 2, and Chen alkamide 2 analogue at 25 microM and by Bauer alkamide 14 at 10 microM. Cytotoxicity did not play a role in the noted reduction of PGE2 production in either the Echinacea extracts or synthesized alkamides. High-performance liquid chromatography analysis identified individual alkamides present at concentrations below 2.8 microM in the extracts from the six Echinacea species (15 microg/mL crude extract). Because active extracts contained Echinacea in a synergistic or additive manner.

  19. Intracellular Trafficking Modulation by Ginsenoside Rg3 Inhibits Brucella abortus Uptake and Intracellular Survival within RAW 264.7 Cells.

    Science.gov (United States)

    Huy, Tran Xuan Ngoc; Reyes, Alisha Wehdnesday Bernardo; Hop, Huynh Tan; Arayan, Lauren Togonon; Min, WonGi; Lee, Hu Jang; Rhee, Man Hee; Chang, Hong Hee; Kim, Suk

    2017-03-28

    Ginsenoside Rg3, a saponin extracted from ginseng, has various pharmacological and biological activities; however, its effects against Brucella infection are still unclear. Herein, the inhibitory effects of ginsenoside Rg3 against intracellular parasitic Brucella infection were evaluated through bacterial infection, adherence assays, and LAMP-1 colocalization, as well as immunoblotting and FACS for detecting MAPK signaling proteins and F-actin polymerization, respectively. The internalization, intracellular growth, and adherence of Brucella abortus in Rg3-treated RAW 264.7 cells were significantly decreased compared with the Rg3-untreated control. Furthermore, an apparent reduction of F-actin content and intensity of F-actin fluorescence in Rg3-treated cells was observed compared with B. abortus -infected cells without treatment by flow cytometry analysis and confocal microscopy, respectively. In addition, treating cells with Rg3 decreased the phosphorylation of MAPK signaling proteins such as ERK 1/2 and p38 compared with untreated cells. Moreover, the colocalization of B. abortus -containing phagosomes with LAMP-1 was markedly increased in Rg3-treated cells. These findings suggest that ginsenoside Rg3 inhibits B. abortus infection in mammalian cells and can be used as an alternative approach in the treatment of brucellosis.

  20. Effect of L-carnitine on the synthesis of nitric oxide in RAW 264·7 murine macrophage cell line.

    Science.gov (United States)

    Koc, A; Ozkan, T; Karabay, A Z; Sunguroglu, A; Aktan, F

    2011-12-01

    L-Carnitine (β-hydroxy-γ-trimethyl aminobutyric acid) plays a critical role in inflammatory diseases by modulating inflammatory cell functions. Inducible nitric oxide synthase (iNOS), a proinflammatory enzyme responsible for the generation of nitric oxide (NO), has been implicated in the pathogenesis of inflammatory diseases. Mechanism of action of L-carnitine on inflammation via iNOS and nuclear factor κB (NF-κB) is unclear. In this study, we aimed to investigate the effect of L-carnitine on nitric oxide synthesis in lipopolysaccharide (LPS)-stimulated RAW 264·7 macrophage cells. For this purpose, cells were pretreated with various concentrations of L-carnitine and subsequently incubated with LPS (1 µg·ml(-1) ). NO levels, iNOS protein expression, and NF-κB activity were determined using colorimetric detection, Western blotting and transfection assays. Our results showed that treatment with L-carnitine suppressed nitric oxide production, iNOS protein expression and NF-κB activity. We demonstrated that inhibitory effect of L-carnitine on iNOS protein expression is at transcriptional level. This study may contribute to understanding the anti-inflammatory effect of L-carnitine. Copyright © 2011 John Wiley & Sons, Ltd.

  1. Capillary zone electrophoresis-multiple reaction monitoring from 100 pg of RAW 264.7 cell lysate digest.

    Science.gov (United States)

    Sun, Liangliang; Li, Yihan; Champion, Matthew M; Zhu, Guijie; Wojcik, Roza; Dovichi, Norman J

    2013-06-07

    Capillary zone electrophoresis-multiple/single reaction monitoring (CZE-MRM/SRM), which employed an electrokinetically driven sheath-flow electrospray interface, was used for the rapid and highly sensitive detection of protein analytes in complex tryptic digests. MRM channels were developed against a commercial exponential mixture of bovine proteins. Five proteins spanning four orders of magnitude concentration range were confidently detected from only 2.5 ng of the digest mixture; the mass detection limits (S/N = 3) of two detected proteins, alpha-casein and glutamate dehydrogenase were about 600 zmol and 30 amol, respectively. This technique was then applied to a RAW 264.7 cell lysate digest. Three proteins were confidently and reproducibly detected from 100 pg of this digest. The sample amount corresponds to the approximate protein content from a single cell, which suggests that CZE-MRM may be a useful analytical tool in chemical cytometry. In addition to providing highly sensitive detection of proteins in complex mixtures, this system is highly rapid; migration time of the protein digests was less than 10 min.

  2. Mycelia extracts of fungal strains isolated from Cordyceps sinensis differently enhance the function of RAW 264.7 macrophages.

    Science.gov (United States)

    Meng, Lan-Zhen; Lin, Bao-Qin; Wang, Bo; Feng, Kun; Hu, De-Jun; Wang, Lan-Ying; Cheong, Kit-Leong; Zhao, Jing; Li, Shao-Ping

    2013-07-30

    Cordyceps sinensis, an entomogenous fungus used in traditional Chinese medicine with multiple pharmacological activities. However, its usage has been limited due to the high price and short supply. Isolate of fungi strains from natural Cordyceps sinensis to achieve a large-scale production by fermentation is an alternative choice. The aim of this study was to investigate and compare the effects of mycelia extracts of different fungal stains isolated from natural Cordyceps sinensis on macrophage functions in vitro. Macrophages' proliferation, phagocytosis, nitric oxide (NO) production, cytokines secretion, iNOS, NF-κB p65 activation and translocation were investigated by the MTT assay, flow cytometry assay, Griess reagent method, ELISA, western blot and immunostaining assay, respectively. The results showed that the effects of cultured Cordyceps mycelia of different fungal strains isolated from natural Cordyceps sinensis on macrophages greatly variant. Among 17 Cordyceps aqueous extracts, only five extracts (UM01, QH11, BNQM, GNCC and DCXC) could significantly increase cell proliferation and NO production of RAW 264.7 mouse macrophages. Moreover, the five extracts, especially UM01 and QH11, significantly enhanced phagocytosis and promoted cytokines release of macrophages. Polysaccharides in cultured UM01 mycelia were found to be the main immune stimulating compounds. The variation of biological effects of fermented mycelia of different fungal strains from natural Cordyceps sinensis may be derived from their chemical diversity, especially polysaccharides, which need further study in future. Copyright © 2013 Elsevier Ireland Ltd. All rights reserved.

  3. The effects of Lycii Radicis Cortex on RANKL-induced osteoclast differentiation and activation in RAW 264.7 cells.

    Science.gov (United States)

    Kim, Jae-Hyun; Kim, Eun-Young; Lee, Bina; Min, Ju-Hee; Song, Dea-Uk; Lim, Jeong-Min; Eom, Ji Whan; Yeom, Mijung; Jung, Hyuk-Sang; Sohn, Youngjoo

    2016-03-01

    Post-menopausal osteoporosis is a serious age-related disease. After the menopause, estrogen deficiency is common, and excessive osteoclast activity causes osteoporosis. Osteoclasts are multinucleated cells generated from the differentiation of monocyte/macrophage precursor cells such as RAW 264.7 cells. The water extract of Lycii Radicis Cortex (LRC) is made from the dried root bark of Lycium chinense Mill. and is termed 'Jigolpi' in Korea. Its effects on osteoclastogenesis and post‑menopausal osteoporosis had not previously been tested. In the present study, the effect of LRC on receptor activator of nuclear factor-κB (NF-κB) ligand (RANKL)-induced osteoclast differentiation was demonstrated using a tartrate-resistant acid phosphatase (TRAP) assay and pit formation assay. Moreover, in order to analyze molecular mechanisms, we studied osteoclastogenesis-related markers such as nuclear factor of activated T-cells cytoplasmic 1 (NFATc1), c-Fos, receptor activator of NF-κB (RANK), TRAP, cathepsin K (CTK), matrix metallopeptidase-9 (MMP-9), calcitonin receptor (CTR) and carbonic anhydrase Ⅱ (CAII) using RT-qPCR and western blot analysis. Additionally, we also determined the effect of LRC on an ovariectomized (OVX) rat model. We noted that LRC inhibited RANKL-induced osteoclast differentiation via suppressing osteoclastogenesis-related markers. It also inhibited osteoporosis in the OVX rat model by decreasing loss of bone density and trabecular area. These results suggest that LRC exerts a positive effect on menopausal osteoporosis.

  4. Perceptual quality estimation of H.264/AVC videos using reduced-reference and no-reference models

    Science.gov (United States)

    Shahid, Muhammad; Pandremmenou, Katerina; Kondi, Lisimachos P.; Rossholm, Andreas; Lövström, Benny

    2016-09-01

    Reduced-reference (RR) and no-reference (NR) models for video quality estimation, using features that account for the impact of coding artifacts, spatio-temporal complexity, and packet losses, are proposed. The purpose of this study is to analyze a number of potentially quality-relevant features in order to select the most suitable set of features for building the desired models. The proposed sets of features have not been used in the literature and some of the features are used for the first time in this study. The features are employed by the least absolute shrinkage and selection operator (LASSO), which selects only the most influential of them toward perceptual quality. For comparison, we apply feature selection in the complete feature sets and ridge regression on the reduced sets. The models are validated using a database of H.264/AVC encoded videos that were subjectively assessed for quality in an ITU-T compliant laboratory. We infer that just two features selected by RR LASSO and two bitstream-based features selected by NR LASSO are able to estimate perceptual quality with high accuracy, higher than that of ridge, which uses more features. The comparisons with competing works and two full-reference metrics also verify the superiority of our models.

  5. Screening of Indonesian medicinal plants for inhibitor activity on nitric oxide production of RAW264.7 cells and antioxidant activity.

    Science.gov (United States)

    Choi, Eun-Mi; Hwang, Jae-Kwan

    2005-03-01

    Traditional Indonesian medicinal plants were screened for their inhibitory effects on the nitric oxide (NO) production in lipopolysaccharide (LPS)-stimulated RAW264.7 macrophages and for the antioxidant activity through the evaluation of free radical scavenging effect and reducing power. The results of screening indicated that 50 methanolic extracts inhibited (>50%) lipopolysaccharides (LPS)-induced NO release from RAW264.7 cells at 50 microg/ml, with 18 having greater than 100% inhibition. At 200 microg/ml, 61 methanol extracts exhibited inhibitory activity (>50%), with 45 showing greater than 100% inhibition. In addition, the free radical scavenging effects of 6 methanolic extracts were found to be more than 50% for extract concentration of 0.5 mug/ml. The results indicate that the extracts contain active compounds that inhibit NO release and scavenge free radical.

  6. Cross-Layer Framework for Multiuser Real Time H.264/AVC Video Encoding and Transmission over Block Fading MIMO Channels Using Outage Probability

    Directory of Open Access Journals (Sweden)

    Slavche Pejoski

    2014-01-01

    Full Text Available We present a framework for cross-layer optimized real time multiuser encoding of video using a single layer H.264/AVC and transmission over MIMO wireless channels. In the proposed cross-layer adaptation, the channel of every user is characterized by the probability density function of its channel mutual information and the performance of the H.264/AVC encoder is modeled by a rate distortion model that takes into account the channel errors. These models are used during the resource allocation of the available slots in a TDMA MIMO communication system with capacity achieving channel codes. This framework allows for adaptation to the statistics of the wireless channel and to the available resources in the system and utilization of the multiuser diversity of the transmitted video sequences. We show the effectiveness of the proposed framework for video transmission over Rayleigh MIMO block fading channels, when channel distribution information is available at the transmitter.

  7. In vitro assesment of anti-inflammatory activities of coumarin and Indonesian cassia extract in RAW264.7 murine macrophage cell line

    Science.gov (United States)

    Sandhiutami, Ni Made Dwi; Moordiani, Moordiani; Laksmitawati, Dian Ratih; Fauziah, Nurul; Maesaroh, Maesaroh; Widowati, Wahyu

    2017-01-01

    Objective(s): Inflammation is an immune response toward injuries. Although inflammation is healing response, but in some condition it will lead to chronic disease such as rheumatoid arthritis, inflammatory bowel disease, atherosclerosis, Alzheimer’s and various cancer. Indonesian cassia (Cinnamomum burmannil C. Nees & T. Ness) known to contain coumarin, is widely used for alternative medicine especially as an anti-inflammatory. This study was conducted to determine the anti-inflammatory properties of coumarin and Indonesian cassia extract (ICE) in LPS-induced RAW264.7 cell line. Materials and Methods: The cytotoxic assay of coumarin and ICE against RAW264.7 cells was conducted using MTS (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium). The anti-inflammatory potential was determined using LPS-induced RAW 267.4 macrophages cells to measure inhibitory activity of compound and ISEon production of nitric oxide (NO), prostaglandin E2 (PGE2), and also cytokines such as interleukin-6 (IL-6), interleukin-1β (IL-1β) and TNF-α. Results: Coumarin 10 µM and ICE 10 µg/ml were nontoxic to the RAW264.7 cells. Both of coumarin and ICE were capable to reduce the PGE2, TNF-α, NO, IL-6, and IL-β level in LPS-induced RAW264.7 cells. Coumarin had higher activity to decrease PGE2 and TNF-α, whilst ICE had higher activity to inhibit NO, IL-6, and IL-β levels. Conclusion: Coumarin and ICE possess anti-inflammatory properties through inhibition of PGE2 and NO along with pro-inflammatory cytokines TNF-α, IL-6, IL-1β production. PMID:28133531

  8. Hexane fraction from Laminaria japonica exerts anti-inflammatory effects on lipopolysaccharide-stimulated RAW 264.7 macrophages via inhibiting NF-kappaB pathway.

    Science.gov (United States)

    Lee, Ji-Young; Lee, Min-Sup; Choi, Hee-Jeon; Choi, Ji-Woong; Shin, Taisun; Woo, Hee-Chul; Kim, Jae-Il; Kim, Hyeung-Rak

    2013-02-01

    Laminaria japonica is a representative marine brown alga used as a culinary item in East Asia. L. japonica extract was shown to exert various biological activities; however, its anti-inflammatory activity has not been reported. The aim of this study is to investigate the molecular mechanisms underlying its anti-inflammatory action. Anti-inflammatory mechanisms of L. japonica n-hexane fraction (LHF) were assessed using lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. An anti-inflammatory compound isolated from LHF by reverse-phase chromatography was identified using nuclear magnetic resonance (NMR) spectroscopy. Our results indicate that LHF significantly inhibited LPS-stimulated nitric oxide (NO) and prostaglandin E(2) (PGE(2)) secretion in a dose-dependent manner and suppressed the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) with no cytotoxicity. As results, levels of pro-inflammatory cytokines were significantly reduced by pretreatment of LHF in LPS-stimulated RAW 264.7 cells. Treatment of LHF strongly suppressed nuclear factor-κB (NF-κB) promoter-driven expression and nuclear translocation of NF-κB by preventing proteolytic degradation of inhibitor of κB (IκB)-α in LPS-stimulated RAW 264.7 cells. Moreover, LHF inhibited the phosphorylation of Akt and mitogen-activated protein kinase (MAPK) in LPS-stimulated RAW 264.7 cells. One of the anti-inflammatory compounds was isolated from LHF and identified as fucoxanthin. These results indicate that the LHF-mediated inhibition of NO and PGE(2) secretion in LPS-stimulated macrophages is regulated by NF-κB inactivation through inhibition of IκB-α, MAPKs, and Akt phosphorylation. LHF may be considered as a functional food candidate for the prevention or treatment of inflammatory diseases.

  9. Interactions between Streptomyces californicus and Stachybotrys chartarum can induce apoptosis and cell cycle arrest in mouse RAW264.7 macrophages

    International Nuclear Information System (INIS)

    Penttinen, Piia; Pelkonen, Jukka; Huttunen, Kati; Toivola, Mika; Hirvonen, Maija-Riitta

    2005-01-01

    Exposure to complex mixtures of bacteria and fungi in moisture-damaged buildings is a potential cause of inflammatory related symptoms among occupants. The present study assessed interactions between two characteristic moldy house microbes Streptomyces californicus and Stachybotrys chartarum. Differences in cytotoxic and inflammatory responses in mouse (RAW264.7) macrophages were studied after exposure to the spores of co-cultivated microbes, the mixture of separately cultivated spores, and the spores of either of these microbes cultivated alone. The RAW264.7 cells were exposed to six doses (1 x 10 4 to 3 x 10 6 spores/ml) for 24 h, and the time course of the induced responses was evaluated after 4, 8, 16, and 24 h of exposure (1 x 10 6 spores/ml). The cytotoxic potential of the spores was characterized by the MTT test, DNA content analysis, and enzyme assay for caspase-3 activity. The production of cytokines (IL-1β, IL-6, IL-10, TNFα, and MIP2) was measured immunochemically and nitric oxide by the Griess method. Co-cultivation increased the ability of the spores to cause apoptosis by more than 4-fold and the proportion of RAW264.7 cells at the G 2 /M stage increased nearly 2-fold when compared to the response induced by the mixture of spores. In contrast, co-cultivation decreased significantly the ability of the spores to trigger the production of NO and IL-6 in RAW264.7 cells. In conclusion, these data suggest that co-culture of S. californicus and S. chartarum can result in microbial interactions that significantly potentiate the ability of the spores to cause apoptosis and cell cycle arrest in mammalian cells

  10. Cytotoxicity and immunomodulatory effects of sol-gel combustion based titanium dioxide (TiO2) particles of large surface area on RAW 264.7 macrophages.

    Science.gov (United States)

    Dinesh, Palani; Suresh Yadav, C; Kannadasan, Sathanandhan; Rasool, Mahaboobkhan

    2017-09-01

    The current study was designed to investigate the cytotoxicity and immunomodulatory effects of sol-gel combustion based TiO 2 particles (glycine and l-alanine as reducing agents) of large surface area on RAW 264.7 macrophages. RAW 264.7 macrophages exposed to varying concentrations of TiO 2 particles (0.001 to 1000μg/ml) were assessed after 24h and showed a reduced cell viability at 100 and 1000μg/ml and increased LDH release at 10μg/ml. Furthermore, TiO 2 particles (0.1, 1 and 10μg/ml) were utilized to assess the immune responses and intracellular ROS levels on RAW 264.7 macrophages. TiO 2 particles at 10μg/ml showed increased mRNA expression of inflammatory cytokines (TNFα, IL-1β and IL-6), inflammatory mediators (iNOS and COX-2) and transcription factor (NFκB) similar to that of LPS stimulated macrophages. However, the mRNA expression levels were found near normal levels at lower concentrations (0.1 and 1μg/ml). In addition, TiO 2 particles at 10μg/ml also increased the production of inflammatory cytokines (TNFα, IL-1β and IL-6) and intracellular ROS levels in RAW 264.7 macrophages similar to that of LPS stimulated macrophages. Conclusively, TiO 2 particles prepared through this method at a concentration≤0.1μg/ml can be used for various biological applications with minimal immunomodulatory effects. Copyright © 2017. Published by Elsevier Ltd.

  11. Extracts of Porphyra tenera (Nori Seaweed) Activate the Immune Response in Mouse RAW264.7 Macrophages via NF-κB Signaling.

    Science.gov (United States)

    Song, Ji-Hye; Kang, Hee-Bum; Park, Seung-Ho; Jeong, Ji-Hoon; Park, Jeongjin; You, Yanghee; Lee, Yoo-Hyun; Lee, Jeongmin; Kim, Eungpil; Choi, Kyung-Chul; Jun, Woojin

    2017-12-01

    Porphyra tenera, also known as nori, is a red algal species of seaweed. It is cultivated in Asia for culinary purposes. We report that P. tenera extract (PTE) enhances the immune response in mouse macrophages. We found that P. tenera extract regulates the NF-κB IκB kinase (IKK) signaling pathway, and we assessed the expression and translocation of p65, a subunit of NF-κB, in RAW264.7 mouse macrophage cells after treatment with PTE. We also investigated the effects of 10% ethanol PTE (PTE10) in RAW264.7 cells. The production of IL-10, IL-6, TNF-α, and IFN-γ was induced by PTE treatment of the macrophages, and PTE also enhanced p-IκB and p-AKT. PTE10 showed no cytotoxicity at 10-20 μg/mL in RAW264.7 cells. PTE10, in fact, increased cell viability at 24 h, stimulated macrophage cells, and induced the phosphorylation of Akt. Akt stimulates IKK activity through the phosphorylation of IKKα and enhances immune activity through the activation of NF-κB. In this study, NF-κB activation was induced by increasing p-NF-κB and p-IKK. A subunit of NF-κB, p65, was located in the nucleus and increased the expression of various cytokines. PTE thus enhanced the immune response through IκB-α immunostimulation signaling in RAW264.7 cells. PTE10 has potential therefore for development of future treatments requiring immune system stimulation.

  12. Pulsed electromagnetic field inhibits RANKL-dependent osteoclastic differentiation in RAW264.7 cells through the Ca2+-calcineurin-NFATc1 signaling pathway.

    Science.gov (United States)

    Zhang, Jie; Xu, Haixia; Han, Zhongyu; Chen, Ping; Yu, Qiang; Lei, Yutian; Li, Zongze; Zhao, Ming; Tian, Jing

    2017-01-08

    Pulsed electromagnetic field (PEMF) has been reported to improve bone healing in osteoporosis patients. However, the precise mechanism has remained largely unknown. This study aimed to investigate the effects of PEMF on nuclear factor κB ligand (RANKL)-dependent osteoclastic differentiation and the Ca 2+ -calcineurin-NFATc1 signaling pathway in RAW264.7 cells in vitro. Treating RAW264.7 cells with RANKL for 4 days induced osteoclastic differentiation in vitro, and the formation of multinucleated osteoclasts, bone resorption-pit formation, tartrate-resistant acid phosphatase (TRAP) activity and the protein levels of cathepsin K, TRAP, Nuclear factor of activated T-cells, cytoplasmic 1 (NFATc1) and matrix metalloproteinase 9 (MMP-9) were significantly decreased. The mRNA levels of specific genes related to osteoclastogenesis (TRAP, NFATc1, CTSK and MMP-9) were also reduced. Moreover, the oscillations of intracellular Ca 2+ in RANKL-dependent RAW264.7 cells were suppressed by PEMF, as well as by inhibitors of membrane and store-operated Ca 2+ channels. Meanwhile, calcineurin activity was increased, although its protein level was not changed. PEMF increased phospho-NFATc1 in the cytosol while suppressing the nuclear translocation of NFATc1, thus inhibiting osteoclastic differentiation by suppressing the Ca 2+ -calcineurin-NFATc1 signaling pathway. Although many questions remain unresolved, to our knowledge, this is the first report demonstrating that PEMF is beneficial against RANKL-dependent osteoclastic differentiation in RAW264.7 cells in vitro via inhibiting the Ca 2+ -calcineurin-NFATc1 signaling pathway. Copyright © 2016 Elsevier Inc. All rights reserved.

  13. Selenium Supplementation of Amaranth Sprouts Influences Betacyanin Content and Improves Anti-Inflammatory Properties via NF?B in Murine RAW 264.7 Macrophages

    OpenAIRE

    Tyszka-Czochara, Malgorzata; Pasko, Pawel; Zagrodzki, Pawel; Gajdzik, Ewelina; Wietecha-Posluszny, Renata; Gorinstein, Shela

    2015-01-01

    Sprouts contain potent compounds which while influencing crucial transduction pathways in cell reveal anti-inflammatory and anticancer activities. In this study, we report the biological activity for seeds and colourful sprouts of four types of edible amaranth, as amaranth has recently attracted interest due to its appreciable nutritional value. MTT assay conducted for the amaranth seeds and sprouts did not show any adverse effect on the viability of murine RAW 264.7 cells. As amaranth accumu...

  14. Immunomodulatory Effect of Flavonoids of Blueberry (Vaccinium corymbosum L.) Leaves via the NF-κB Signal Pathway in LPS-Stimulated RAW 264.7 Cells

    Science.gov (United States)

    Shi, Dazhi; Xu, Mengyi; Pan, Enshan; Luo, Chaohua

    2017-01-01

    Objective This study aimed to explore the immunoregulatory effect of flavonoids of blueberry (Vaccinium corymbosum L.) leaves (FBL). Methods The flavonoids of blueberry leaves were prepared with 70% ethanol and were identified by ultraperformance liquid chromatography/quadrupole-time-of-flight mass spectrometry (UPLC/Q-Tof-MS). The immunoregulatory effect and possible regulatory mechanisms of FBL were investigated in lipopolysaccharide- (LPS-) induced RAW 264.7 cells. Results According to the results of UPLC/Q-Tof-MS, nine flavonoids of blueberry leaves were identified. FBL showed a significant reduction in the production of TNF-α in LPS-stimulated RAW 264.7 cells. FBL significantly decreased the expression of NF-κB p65 and P-NF-κB p65 in LPS-induced RAW 264.7 cells in a dose-dependent manner. Conclusion Our study showed the immunoregulatory effect of FBL through the suppression of TNF-α via the NF-κB signal pathway. PMID:29445755

  15. Purification and Structural Characterization of a Novel Water-Soluble Neutral Polysaccharide from Cantharellus cibarius and Its Immunostimulating Activity in RAW264.7 Cells

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    Long Chen

    2017-01-01

    Full Text Available Polysaccharide is one of the important active ingredients of Cantharellus cibarius. The aims of this work were to analyze preliminary characterization and to investigate immunostimulating activity of a novel water-soluble neutral polysaccharide named JP1, which was purified from the fruiting body of Cantharellus cibarius using DEAE-FF chromatography and Sephadex G-100 chromatography. The characteristics of JP1 were determined by HPGPC, FT-IR spectra, gas chromatography, and Congo Red Method. Immunostimulating activity of JP1 was investigated in RAW264.7 cells. Results indicated that JP1 consisted of L-Arabinose, D-Mannose, D-Glucose, and D-Galactose in a molar ratio of 1 : 1.06 : 1.95 : 1.17 with a molecular weight of 336 kDa. JP1 is nontoxic to RAW264.7 cells at this concentration range (62.5–1000 μg/mL. Furthermore, JP1 can promote mouse peritoneal macrophages to secrete NO and enhance the secretion of macrophages’ cytokines IL-6 in RAW264.7 cells. These results suggested that JP1 could have potential immunostimulating activity applications as medicine or functional food.

  16. Andrographolide Inhibits Inflammatory Cytokines Secretion in LPS-Stimulated RAW264.7 Cells through Suppression of NF-κB/MAPK Signaling Pathway

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    Yu Li

    2017-01-01

    Full Text Available Andrographolide, the main active component extracted from Andrographis paniculata (Burm.f. Wall. ex Nees, exerts anti-inflammatory effects; however, the principal molecular mechanisms remain unclear. The objective of this study was to investigate the molecular mechanisms of Andrographolide in modifying lipopolysaccharide- (LPS- induced signaling pathway in RAW264.7 cells. An in vitro model of inflammation was induced by LPS in mouse RAW264.7 cells in the presence of Andrographolide. The concentration and expression levels of proinflammatory cytokines were determined by an enzyme-linked immunosorbent assay (ELISA and quantitative real-time polymerase chain reaction (qRT-PCR, respectively. The nuclear level of NF-κB was measured by an electrophoretic mobility shift assay (EMSA. The expression levels of NF-κB, p38, ERK, and JNK were determined by western blot. Andrographolide dose-dependently inhibited the release and mRNA expression of TNF-α, IL-6, and IL-1β in LPS-stimulated RAW264.7 cells. The nuclear level of p65 protein was decreased in Andrographolide treatment group. Western blot analysis showed that Andrographolide suppressed LPS-induced NF-κB activation and the phosphorylation of IkBa, ERK1/2, JNK, and p38. These results suggest that Andrographolide exerts an anti-inflammatory effect by inhibiting the activation of NF-κB/MAPK signaling pathway and the induction of proinflammatory cytokines.

  17. Andrographolide Inhibits Inflammatory Cytokines Secretion in LPS-Stimulated RAW264.7 Cells through Suppression of NF-κB/MAPK Signaling Pathway.

    Science.gov (United States)

    Li, Yu; He, Shengnan; Tang, Jishun; Ding, Nana; Chu, Xiaoyan; Cheng, Lianping; Ding, Xuedong; Liang, Ting; Feng, Shibin; Rahman, Sajid Ur; Wang, Xichun; Wu, Jinjie

    2017-01-01

    Andrographolide, the main active component extracted from Andrographis paniculata (Burm.f.) Wall. ex Nees, exerts anti-inflammatory effects; however, the principal molecular mechanisms remain unclear. The objective of this study was to investigate the molecular mechanisms of Andrographolide in modifying lipopolysaccharide- (LPS-) induced signaling pathway in RAW264.7 cells. An in vitro model of inflammation was induced by LPS in mouse RAW264.7 cells in the presence of Andrographolide. The concentration and expression levels of proinflammatory cytokines were determined by an enzyme-linked immunosorbent assay (ELISA) and quantitative real-time polymerase chain reaction (qRT-PCR), respectively. The nuclear level of NF- κ B was measured by an electrophoretic mobility shift assay (EMSA). The expression levels of NF- κ B, p38, ERK, and JNK were determined by western blot. Andrographolide dose-dependently inhibited the release and mRNA expression of TNF- α , IL-6, and IL-1 β in LPS-stimulated RAW264.7 cells. The nuclear level of p65 protein was decreased in Andrographolide treatment group. Western blot analysis showed that Andrographolide suppressed LPS-induced NF- κ B activation and the phosphorylation of IkBa, ERK1/2, JNK, and p38. These results suggest that Andrographolide exerts an anti-inflammatory effect by inhibiting the activation of NF- κ B/MAPK signaling pathway and the induction of proinflammatory cytokines.

  18. Immunomodulatory Effect of Flavonoids of Blueberry (Vaccinium corymbosum L. Leaves via the NF-κB Signal Pathway in LPS-Stimulated RAW 264.7 Cells

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    Dazhi Shi

    2017-01-01

    Full Text Available Objective. This study aimed to explore the immunoregulatory effect of flavonoids of blueberry (Vaccinium corymbosum L. leaves (FBL. Methods. The flavonoids of blueberry leaves were prepared with 70% ethanol and were identified by ultraperformance liquid chromatography/quadrupole-time-of-flight mass spectrometry (UPLC/Q-Tof-MS. The immunoregulatory effect and possible regulatory mechanisms of FBL were investigated in lipopolysaccharide- (LPS- induced RAW 264.7 cells. Results. According to the results of UPLC/Q-Tof-MS, nine flavonoids of blueberry leaves were identified. FBL showed a significant reduction in the production of TNF-α in LPS-stimulated RAW 264.7 cells. FBL significantly decreased the expression of NF-κB p65 and P-NF-κB p65 in LPS-induced RAW 264.7 cells in a dose-dependent manner. Conclusion. Our study showed the immunoregulatory effect of FBL through the suppression of TNF-α via the NF-κB signal pathway.

  19. Detection of Double-Compressed H.264/AVC Video Incorporating the Features of the String of Data Bits and Skip Macroblocks

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    Heng Yao

    2017-12-01

    Full Text Available Today’s H.264/AVC coded videos have a high quality, high data-compression ratio. They also have a strong fault tolerance, better network adaptability, and have been widely applied on the Internet. With the popularity of powerful and easy-to-use video editing software, digital videos can be tampered with in various ways. Therefore, the double compression in the H.264/AVC video can be used as a first step in the study of video-tampering forensics. This paper proposes a simple, but effective, double-compression detection method that analyzes the periodic features of the string of data bits (SODBs and the skip macroblocks (S-MBs for all I-frames and P-frames in a double-compressed H.264/AVC video. For a given suspicious video, the SODBs and S-MBs are extracted for each frame. Both features are then incorporated to generate one enhanced feature to represent the periodic artifact of the double-compressed video. Finally, a time-domain analysis is conducted to detect the periodicity of the features. The primary Group of Pictures (GOP size is estimated based on an exhaustive strategy. The experimental results demonstrate the efficacy of the proposed method.

  20. Koumine Attenuates Lipopolysaccaride-Stimulated Inflammation in RAW264.7 Macrophages, Coincidentally Associated with Inhibition of NF-κB, ERK and p38 Pathways

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    Zhihang Yuan

    2016-03-01

    Full Text Available Medicinal herbal plants have been commonly used for intervention of different diseases and health enhancement worldwide. Koumine, an alkaloid monomer found abundantly in Gelsemium plants, can be effectively used as an anti-inflammatory medication. In this study, the mechanisms associated with the preventative effect of koumine on lipopolysaccharide (LPS-mediated inflammation in RAW264.7 macrophages were investigated. Koumine induced a decrease in the level of inducible nitric oxide synthase (iNOS protein, concomitant reduction in the production of nitric oxide (NO and reduction of the levels of interleukin (IL-6, tumor necrosis factor-α (TNF-α and IL-1β. Furthermore, koumine decreased the phosphorylation of p65 and inhibited nuclear factor κ Bα (IκBα proteins, resulting in lower production of nuclear factor (NF-κB transactivation. Koumine also induced a decrease in the phosphorylation of extracellular-signal-regulated kinases (ERK and p38 in RAW264 cells. In conclusion, these findings reveal that koumine decreases the productions of pro-inflammatory mediators though the suppression of p38 and ERK MAPK phosphorylation and the inhibition of NF-κB activation in RAW264.7 cells.

  1. Thymus vulgaris L. and thymol assist murine macrophages (RAW 264.7) in the control of in vitro infections by Staphylococcus aureus, Pseudomonas aeruginosa, and Candida albicans.

    Science.gov (United States)

    de Oliveira, Jonatas Rafael; Figueira, Leandro Wagner; Sper, Fábia Lugli; Meccatti, Vanessa Marques; Camargo, Samira Esteves Afonso; de Oliveira, Luciane Dias

    2017-08-01

    Microorganisms are capable to combat defense cells by means of strategies that contribute to their stabilization and proliferation in invaded tissues. Frequently antimicrobial-resistant strains appear; therefore, alternative methods to control them must be investigated, for example, the use of plant products. The capacity of the thyme extract (Thymus vulgaris L.) and phytocompound thymol in the control of in vitro infections by Staphylococcus aureus, Pseudomonas aeruginosa, and Candida albicans in murine macrophages (RAW 264.7) was evaluated. Minimal inhibitory concentrations (MIC) of the plant products were used. The effect of these MIC were analyzed in the assays of phagocytosis and immunoregulation by analysis of the production of cytokines (IL-1β, TNF-α, and IL-10) and nitric oxide (NO). The plant products effectively assisted the macrophages in the phagocytosis of microorganisms, presenting significant reductions of S. aureus and P. aeruginosa. The macrophages also regulated the production of inflammatory mediators in the infections by S. aureus, P. aeruginosa, and C. albicans. In addition, thyme provided a satisfactory effect in response to the bacterial infections, regarding generation of NO. Thus, the effectiveness of the thyme and thymol to control in vitro infections by S. aureus, P. aeruginosa, and C. albicans was observed. Phagocytosis of S. aureus by RAW 264.7 was enhanced with thymol Thyme enhanced the phagocytosis of P. aeruginosa by RAW 264.7 Plant products provided immunoregulation of inflammatory cytokines Production of nitric oxide was improved with the treatments in bacterial infections.

  2. Glycine tomentella Hayata inhibits IL-1β and IL-6 production, inhibits MMP-9 activity, and enhances RAW264.7 macrophage clearance of apoptotic cells

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    Sun Yu-Shu

    2010-11-01

    Full Text Available Abstract Background To assess the effects of Glycine tomentella Hayata (GTH, a traditional herbal medicine for treatment of rheumatic diseases on the expression of the proinflammatory cytokines and on the clearance of apoptotic cells by macrophages. Methods RAW264.7 cells were cultured with lipopolysaccharide (LPS in the presence or absence of ethanol extract of GTH. The expression of proinflammatory cytokines IL-1β, IL-6, and TNF-α, and inducible nitric oxide synthase (iNOS and transglutaminase 2 (TG2 were assayed by reverse transcriptase-polymerase chain reaction (RT-PCR and enzyme-linked immunosorbent assay (ELISA. Matrix metalloproteinase (MMP-2 and MMP-9 were assayed by gelatin zymography. For detecting uptake of apoptotic cells, RAW264.7 cells were cultured with carboxyfluorescein diacetate (CFDA-stained apoptotic cells and assayed by flow cytometry. Results The major components of GTH analyzed by high-performance liquid chromatography (HPLC chromatogram were daidzein (42.5%, epicatechin (28.8%, and naringin (9.4%. GTH treatment inhibited the expression of proinflammatory cytokines IL-1β, IL-6 and MMP-9 but did not affect the expression of TNF-α and iNOS. GTH significantly enhanced the expression of TG2 and the clearance of apoptotic cells by RAW264.7 macrophages. Conclusions GTH inhibits proinflammatory cytokine secretion and MMP-9 activity, enhances apoptotic cell uptake and up-regulates TG2 expression. Our data show that GTH might have beneficial effects on rheumatic diseases.

  3. Morita-Baylis-Hillman Adducts Display Anti-Inflammatory Effects by Modulating Inflammatory Mediator Expression in RAW264.7 Cells

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    Glaucia V. Faheina-Martins

    2017-01-01

    Full Text Available Inflammatory response plays an important role not only in the normal physiology but also in pathologies such as cancers. The Morita-Baylis-Hillman adducts (MBHA are a novel group of synthetic molecules that have demonstrated many biological activities against some parasitic cells such as Plasmodium falciparum, Leishmania amazonensis, and Leishmania chagasi, and antimitotic activity against sea urchin embryonic cells was also related. However, little is known about the mechanisms induced by MBHA in inflammatory process and its relation with anticancer activity. The present work investigated the cytotoxicity of three MBHA derivatives (A2CN, A3CN, and A4CN, on human colorectal adenocarcinoma, HT-29 cells, and their anti-inflammatory activities were examined in lipopolysaccharide- (LPS- stimulated RAW264.7 macrophage cells, being these derivatives potentially cytotoxic to HT-29 cells. Coincubation with A2CN, A3CN, or A4CN and LPS in RAW264.7 cells inhibited NO production, as well as the production of reactive oxygen species (ROS was also repressed. The mRNA expressions of IL-1β and IL-6 were significantly downregulated by such MBHA compounds in RAW264.7 cells, but only A2CN was able to inhibit the COX-2 gene expression. We also showed that MBHA compounds decreased almost to zero the production of IL-1β and IL-6. These findings display that such MBHA compounds exhibit anticancer and anti-inflammatory activities.

  4. Morita-Baylis-Hillman Adducts Display Anti-Inflammatory Effects by Modulating Inflammatory Mediator Expression in RAW264.7 Cells.

    Science.gov (United States)

    Faheina-Martins, Glaucia V; Leite, Jacqueline Alves; Dantas, Bruna Braga; Lima-Júnior, Cláudio G; Vasconcellos, Mário L A A; Rodrigues-Mascarenhas, Sandra; Araújo, Demetrius A M

    2017-01-01

    Inflammatory response plays an important role not only in the normal physiology but also in pathologies such as cancers. The Morita-Baylis-Hillman adducts (MBHA) are a novel group of synthetic molecules that have demonstrated many biological activities against some parasitic cells such as Plasmodium falciparum , Leishmania amazonensis , and Leishmania chagasi , and antimitotic activity against sea urchin embryonic cells was also related. However, little is known about the mechanisms induced by MBHA in inflammatory process and its relation with anticancer activity. The present work investigated the cytotoxicity of three MBHA derivatives (A2CN, A3CN, and A4CN), on human colorectal adenocarcinoma, HT-29 cells, and their anti-inflammatory activities were examined in lipopolysaccharide- (LPS-) stimulated RAW264.7 macrophage cells, being these derivatives potentially cytotoxic to HT-29 cells. Coincubation with A2CN, A3CN, or A4CN and LPS in RAW264.7 cells inhibited NO production, as well as the production of reactive oxygen species (ROS) was also repressed. The mRNA expressions of IL-1 β and IL-6 were significantly downregulated by such MBHA compounds in RAW264.7 cells, but only A2CN was able to inhibit the COX-2 gene expression. We also showed that MBHA compounds decreased almost to zero the production of IL-1 β and IL-6. These findings display that such MBHA compounds exhibit anticancer and anti-inflammatory activities.

  5. Low Molecular Weight Protein Tyrosine Phosphatase Slow Isoform Knockdown in MDA-MB-435 Cells Decreases RAW 264.7 Osteoclastic Differentiation.

    Science.gov (United States)

    Alho, Irina; Costa, Luis; Bicho, Manuel; Coelho, Constança

    2016-05-01

    During the bone metastatic process, tumor cells and bone cells drive a vicious cycle stimulating growth and activity of each other. We here address how low molecular weight protein tyrosine phosphatase (LMW-PTP) could be involved in this process. We targeted LMW-PTP by siRNA and evaluated the effect of various soluble factors released to the culture medium by the MDA-MB-435 breast cancer cell line, in RAW 264.7 osteoclastogenesis. We showed that these soluble factors did not change RAW 264.7 osteoclastogenic potential. The knockdown of the LMW-PTP slow isoform decreased osteoclastogenesis of RAW 264.7, showing less active Src. The knockdown of LMW-PTP and its slow isoform decreased the release of IL-8 but not IL-6 in MDA-MB-435. The LMW-PTP slow isoform can be an important protein in bone metastatic disease, with a fundamental role in the interplay between tumor cells and osteoclasts, through the regulation of Src activity and IL-8 secretion. Copyright© 2016 International Institute of Anticancer Research (Dr. John G. Delinassios), All rights reserved.

  6. Comparison of Anti-Inflammatory Effects of Flavonoid-Rich Common and Tartary Buckwheat Sprout Extracts in Lipopolysaccharide-Stimulated RAW 264.7 and Peritoneal Macrophages

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    Tae Gyu Nam

    2017-01-01

    Full Text Available Buckwheat sprouts have been widely consumed all around world due to their great abundance of bioactive compounds. In this study, the anti-inflammatory effects of flavonoid-rich common buckwheat sprout (CBS and tartary buckwheat sprout (TBS extracts were evaluated in lipopolysaccharide- (LPS- stimulated RAW 264.7 murine macrophages and primary peritoneal macrophages from male BALB/c mice. Based on the reversed-phase HPLC analysis, the major flavonoids in CBS were determined to be C-glycosylflavones (orientin, isoorientin, vitexin, and isovitexin, quercetin-3-O-robinobioside, and rutin, whereas TBS contained only high amounts of rutin. The TBS extract exhibited higher inhibitory activity as assessed by the production of proinflammatory mediators such as nitric oxide and cytokines including tumor necrosis factor-α, interleukin- (IL- 6, and IL-12 in LPS-stimulated RAW 264.7 macrophages than CBS extract. In addition, TBS extract suppressed nuclear factor-kappa B activation by preventing inhibitor kappa B-alpha degradation and mitogen-activated protein kinase phosphorylation in LPS-stimulated RAW 264.7 macrophages. Moreover, the TBS extract markedly reduced LPS-induced cytokine production in peritoneal macrophages. Taken together, these findings suggest that TBS extract can be a potential source of anti-inflammatory agents that may influence macrophage-mediated inflammatory disorders.

  7. Anti-inflammatory effect of tribulusamide D isolated from Tribulus terrestris in lipopolysaccharide-stimulated RAW264.7 macrophages

    Science.gov (United States)

    Lee, Hyun Hwa; Ahn, Eun-Kyung; Hong, Seong-Su; Oh, Joa Sub

    2017-01-01

    Tribulus terrestris (T. terrestris) has been used as a traditional medicine for the treatment of a variety of diseases, including inflammation, edema and hypertension. The aqueous and ethanol extracts of T. terrestris contain alkaloids, flavonoids, tannins, quinines and phenolic compounds. Tribulusamide D is a compound that has been isolated from the ethanol extract of T. terrestris. The present study investigated the anti-inflammatory effect of tribulusamide D on lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. Tribulusamide D inhibited the production of LPS-induced nitric oxide and prostaglandin E2, by reducing the expression of inducible nitric oxide synthase and cyclooxygenase-2 expression, respectively. The expression of these genes associated with inflammation was determined using reverse transcription-polymerase chain reaction and western blot analysis. Furthermore, tribulusamide D reduced the expression of LPS-induced inflammatory cytokines, including interleukin (IL)-6, IL-10 and tumor necrosis factor-α. They were quantified using an enzyme-linked immunosorbent assay. In addition, the present study confirmed that the inhibitory effects of tribulusamide D on the inflammatory response were mediated through inactivation of mitogen-activated protein kinase p38 and inhibition of nuclear localization of nuclear factor-B, which were also determined by western blot analysis. To the best of our knowledge, the current study is the first to demonstrate that tribulusamide D exerts anti-inflammatory activity by altering the expression of inflammatory mediators and cytokines, indicating that tribulusamide D could be developed as a potential therapeutic agent for the treatment of inflammatory disorders. PMID:28849109

  8. Anti-inflammatory effect of Taraxacum officinale leaves on lipopolysaccharide-induced inflammatory responses in RAW 264.7 cells.

    Science.gov (United States)

    Koh, Yoon-Jeoung; Cha, Dong-Soo; Ko, Je-Sang; Park, Hyun-Jin; Choi, Hee-Don

    2010-08-01

    To investigate the efficacy and the mechanism of the anti-inflammatory effect of Taraxacum officinale leaves (TOLs), the effect of a methanol extract and its fractions recovered from TOLs on lipopolysaccharide (LPS)-induced responses was studied in the mouse macrophage cell line, RAW 264.7. Cells were pretreated with various concentrations of the methanol extract and its fractions and subsequently incubated with LPS (1 microg/mL). The levels of nitric oxide (NO), prostaglandin (PG) E(2), and pro-inflammatory cytokines including tumor necrosis factor (TNF)-alpha, interleukin (IL)-1beta, and IL-6 were determined using enzyme-linked immunosorbent assays. Expressions of inducible NO synthase (iNOS) and cyclooxygenase (COX)-2 and activation of mitogen-activated protein (MAP) kinases were analyzed using western blotting. The methanol extract and its fractions inhibited LPS-induced production of NO, pro-inflammatory cytokines, and PGE(2) in a dose-dependent manner. The chloroform fraction significantly suppressed production of NO, PGE(2), and two pro-inflammatory cytokines (TNF-alpha and IL-1beta) in a dose-dependent manner with 50% inhibitory concentration values of 66.51, 90.96, 114.76, and 171.06 microg/mL, respectively. The ethyl acetate fraction also inhibited production of the inflammatory molecules. The chloroform and ethyl acetate fractions reduced LPS-induced expressions of iNOS and COX-2 and activation of MAP kinases in a dose-dependent manner. Among the fractions of the methanol extract, the chloroform and ethyl acetate fractions exhibited the most effective anti-inflammatory activities. These results show that the anti-inflammatory effects of TOLs are probably due to down-regulation of NO, PGE(2), and pro-inflammatory cytokines and reduced expressions of iNOS and COX-2 via inactivation of the MAP kinase signal pathway.

  9. Cytoprotective and enhanced anti-inflammatory activities of liposomal piroxicam formulation in lipopolysaccharide-stimulated RAW 264.7 macrophages

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    Chiong HS

    2013-03-01

    Full Text Available Hoe Siong Chiong,1 Yoke Keong Yong,1 Zuraini Ahmad,1 Mohd Roslan Sulaiman,1 Zainul Amiruddin Zakaria,1 Kah Hay Yuen,2 Muhammad Nazrul Hakim1,31Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Serdang, Malaysia; 2School of Pharmaceutical Sciences, Universiti Sains Malaysia, Gelugor, Malaysia; 3Sports Academy, Universiti Putra Malaysia, Serdang, MalaysiaBackground: Liposomal drug delivery systems, a promising lipid-based nanoparticle technology, have been known to play significant roles in improving the safety and efficacy of an encapsulated drug.Methods: Liposomes, prepared using an optimized proliposome method, were used in the present work to encapsulate piroxicam, a widely prescribed nonsteroidal anti-inflammatory drug. The cytotoxic effects as well as the in vitro efficacy in regulation of inflammatory responses by free-form piroxicam and liposome-encapsulated piroxicam were evaluated using a lipopolysaccharide-sensitive macrophage cell line, RAW 264.7.Results: Cells treated with liposome-encapsulated piroxicam demonstrated higher cell viabilities than those treated with free-form piroxicam. In addition, the liposomal piroxicam formulation resulted in statistically stronger inhibition of pro-inflammatory mediators (ie, nitric oxide, tumor necrosis factor-α, interleukin-1β, and prostaglandin E2 than piroxicam at an equivalent dose. The liposome-encapsulated piroxicam also caused statistically significant production of interleukin-10, an anti-inflammatory cytokine.Conclusion: This study affirms the potential of a liposomal piroxicam formulation in reducing cytotoxicity and enhancing anti-inflammatory responses in vitro.Keywords: liposomes, nitric oxide, cytokines, prostaglandin E2, interleukin-1β, piroxicam

  10. Correlation Analysis on Agronomic Characters in F1 population derived from a cross of Pongsu Seribu 2 and MR 264

    International Nuclear Information System (INIS)

    Nor Aishah, H.; Abdul Rahim Harun; Norain, M.N.

    2014-01-01

    Agronomic characters play a vital role in grain yield performance. However, the information related to the relationships between agronomic characters and grain yield in Malaysia are particularly very limited. Therefore, this study was conducted to determine the correlation between grain yield performance with nine (n=9) agronomic characters namely plant height, panicle length, total grain number, filled grains per panicle, grain length, number of tillers, flag leaf area, flag leaf width and 1000-grains weight. Twenty (n=20) F 1 population derived from a cross of Pongsu Seribu 2 and MR 264 were used and were grown at Malaysian Nuclear Agency Greenhouse, Bangi. Result of correlation analysis revealed a positive and significant at (p ≤ 0.01) relation between 1000-grains weight with total grains number (r = 0.647), filled grains per panicle (r = 0.603), grain length (r = 0.834), flag leaf area (r = 0.701) and flag leaf width (r = 0.894). However, the negative (inverse) correlation was observed between panicle length with total grain number, number of tiller and filled grain per panicle. Regression analysis revealed that flag leaf width (4.162) has highest direct effect on the yield followed by grain length (2.801), flag leaf area (0.048) and total grain no (0.07) respectively. Whereas, plant height (-0.05), plant length (-0.223), filled grains per panicle (-0.005) and number of tillers (-0.071) has indirect effect on the yield. This study indicated that selection based flag leaf width and grain length will be highly effective for yield improvement can be consider in selection of breeding lines and highly effective for yield improvement in rice breeding program. (author)

  11. Rhamnolipids from non-pathogenic Burkholderia thailandensis E264: Physicochemical characterization, antimicrobial and antibiofilm efficacy against oral hygiene related pathogens.

    Science.gov (United States)

    Elshikh, Mohamed; Funston, Scott; Chebbi, Alif; Ahmed, Syed; Marchant, Roger; Banat, Ibrahim M

    2017-05-25

    Biosurfactants are naturally occurring surface active compounds that have mainly been exploited for environmental applications and consumer products, with their biomedical efficacy an emerging area of research. Rhamnolipids area major group of biosurfactants that have been reported for their antimicrobial and antibiofilm efficacy. One of the main limiting factors for scaled up production and downstream applications of rhamnolipids is the fact that they are predominantly produced from the opportunistic pathogen Pseudomonas aeruginosa. In this article, we have reported the production and characterisation of long chain rhamnolipids from non-pathogenic Burkholderia thailandensis E264 (ATCC 700388). We have also investigated the antibacterial and antibiofilm properties of these rhamnolipids against some oral pathogens (Streptococcus oralis, Actinomyces naeslundii, Neisseria mucosa and Streptococcus sanguinis), important for oral health and hygiene. Treating these bacteria with different concentrations of long chain rhamnolipids resulted in a reduction of 3-4 log of bacterial viability, placing these rhamnolipids close to being classified as biocidal. Investigating long chain rhamnolipid efficacy as antibiofilm agents for prospective oral-related applications revealed good potency against oral-bacteria biofilms in a co-incubation experiments, in a pre-coated surface format, in disrupting immature biofilms and has shown excellent combination effect with Lauryl Sodium Sulphate which resulted in a drastic decrease in its minimal inhibitory concentration against different bacteria. Investigating the rhamnolipid permeabilization effect along with their ability to induce the formation of reactive oxygen species has shed light on the mechanism through which inhibition/killing of bacteria may occur. Copyright © 2017 Elsevier B.V. All rights reserved.

  12. The Effect of Bee Venom on COX-2, P38, ERK and JNK in RAW 264.7 Cells

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    Jae-Young Sim

    2003-06-01

    Full Text Available Objectives : The purpose of this study was to investigate the effect of Bee Venom on the lipopolysaccharide(LPS, sodium nitroprusside(SNP, hydrogen peroxide(H2O2-induced expressions of cyclooxygenase-2(COX-2, p38, jun N-terminal Kinase(JNK and extra-signal response kinase(ERK in RAW 264.7 cells, a murine macrophage cell line. Methods : The expressions of COX-2, p38, JNK and ERK were determined by western blotting with corresponding antibodies.\\ Results : 1. The 0.5, 1 and 5 ㎍/㎖ of bee venom inhibited significantly LPS and SNP-induced expression of COX-2 compared with control, respectively. The 0.5, 1 and 5 ㎍/㎖ of bee venom inhibited insignificantly H2O2-induced expression of COX-2 compared with control, respectively. 2. The 0.5, 1 and 5 ㎍/㎖ of bee venom inhibited significantly LPS, SNP and H2O2-induced expression of p38 compared with control, respectively. 3. The 1 and 5 ㎍/㎖ of bee venom inhibited significantly SNP-induced expression of JNK compared with control, respectively. All of bee venom inhibited insignificantly LPS and H2O2-induced expression of JNK compared with control, respectively. 4. The 5 ㎍/㎖ of bee venom inhibited significantly SNP-induced expression of ERK, the 0.5 ㎍/㎖ of bee venom increased significantly H2O2-induced expression of ERK compared with control. The 0.5, 1 and 5 ㎍/㎖ of bee venom inhibited insignificantly LPS-induced expression of ERK compared with control, respectively.

  13. Ascorbic acid-pretreated quartz enhances cyclo-oxygenase-2 expression in RAW 264.7 murine macrophages.

    Science.gov (United States)

    Scarfì, Sonia; Benatti, Umberto; Pozzolini, Marina; Clavarino, Emanuela; Ferraris, Chiara; Magnone, Mirko; Valisano, Laura; Giovine, Marco

    2007-01-01

    Exposure to quartz particles induces a pathological process named silicosis. Alveolar macrophages initiate the disease through their activation, which is the origin of the later dysfunctions. Ascorbic acid is known to selectively dissolve the quartz surface. During the reaction, ascorbic acid progressively disappears and hydroxyl radicals are generated from the quartz surface. These observations may be relevant to mammalian quartz toxicity, as substantial amounts of ascorbic acid are present in the lung epithelium. We studied the inflammatory response of the murine macrophage cell line RAW 264.7 incubated with ascorbic acid-treated quartz, through the expression and activity of the enzyme cyclo-oxygenase-2 (COX-2). COX-2 expression and prostaglandin secretion were enhanced in cells incubated with ascorbic acid-treated quartz. In contrast, no changes were observed in cells incubated with Aerosil OX50, an amorphous form of silica. Quantification of COX-2 mRNA showed a threefold increase in cells incubated with ascorbic acid-treated quartz compared with controls. The transcription factors, NF-kappaB, pCREB and AP-1, were all implicated in the increased inflammatory response. Reactive oxygen species (H(2)O(2) and OH(*)) were involved in COX-2 expression in this experimental model. Parallel experiments performed on rat alveolar macrophages from bronchoalveolar lavage confirmed the enhanced COX-2 expression and activity in the cells incubated with ascorbic acid-treated quartz compared with untreated quartz. In conclusion, the selective interaction with, and modification of, quartz particles by ascorbic acid may be a crucial event determining the inflammatory response of macrophages, which may subsequently develop into acute inflammation, eventually leading to the chronic pulmonary disease silicosis.

  14. Is nitric oxide decrease observed with naphthoquinones in LPS stimulated RAW 264.7 macrophages a beneficial property?

    Directory of Open Access Journals (Sweden)

    Brígida R Pinho

    Full Text Available The search of new anti-inflammatory drugs has been a current preoccupation, due to the need of effective drugs, with less adverse reactions than those used nowadays. Several naphthoquinones (plumbagin, naphthazarin, juglone, menadione, diosquinone and 1,4-naphthoquinone, plus p-hydroquinone and p-benzoquinone were evaluated for their ability to cause a reduction of nitric oxide (NO production, when RAW 264.7 macrophages were stimulated with lipopolysaccharide (LPS. Dexamethasone was used as positive control. Among the tested compounds, diosquinone was the only one that caused a NO reduction with statistical importance and without cytotoxicity: an IC(25 of 1.09±0.24 µM was found, with 38.25±6.50% (p<0.001 NO reduction at 1.5 µM. In order to elucidate if this NO decrease resulted from the interference of diosquinone with cellular defence mechanisms against LPS or to its conversion into peroxynitrite, by reaction with superoxide radical formed by naphthoquinones redox cycling, 3-nitrotyrosine and superoxide determination was also performed. None of these parameters showed significant changes relative to control. Furthermore, diosquinone caused a decrease in the pro-inflammatory cytokines: tumour necrosis factor-alpha (TNF-α and interleukin 6 (IL-6. Therefore, according to the results obtained, diosquinone, studied for its anti-inflammatory potential for the first time herein, has beneficial effects in inflammation control. This study enlightens the mechanisms of action of naphthoquinones in inflammatory models, by checking for the first time the contribution of oxidative stress generated by naphthoquinones to NO reduction.

  15. Anti-inflammatory effect of tribulusamide D isolated from Tribulus terrestris in lipopolysaccharide-stimulated RAW264.7 macrophages.

    Science.gov (United States)

    Lee, Hyun Hwa; Ahn, Eun-Kyung; Hong, Seong-Su; Oh, Joa Sub

    2017-10-01

    Tribulus terrestris (T. terrestris) has been used as a traditional medicine for the treatment of a variety of diseases, including inflammation, edema and hypertension. The aqueous and ethanol extracts of T. terrestris contain alkaloids, flavonoids, tannins, quinines and phenolic compounds. Tribulusamide D is a compound that has been isolated from the ethanol extract of T. terrestris. The present study investigated the anti‑inflammatory effect of tribulusamide D on lipopolysaccharide (LPS)‑stimulated RAW 264.7 macrophages. Tribulusamide D inhibited the production of LPS‑induced nitric oxide and prostaglandin E2, by reducing the expression of inducible nitric oxide synthase and cyclooxygenase‑2 expression, respectively. The expression of these genes associated with inflammation was determined using reverse transcription‑polymerase chain reaction and western blot analysis. Furthermore, tribulusamide D reduced the expression of LPS‑induced inflammatory cytokines, including interleukin (IL)‑6, IL‑10 and tumor necrosis factor‑α. They were quantified using an enzyme‑linked immunosorbent assay. In addition, the present study confirmed that the inhibitory effects of tribulusamide D on the inflammatory response were mediated through inactivation of mitogen‑activated protein kinase p38 and inhibition of nuclear localization of nuclear factor‑B, which were also determined by western blot analysis. To the best of our knowledge, the current study is the first to demonstrate that tribulusamide D exerts anti‑inflammatory activity by altering the expression of inflammatory mediators and cytokines, indicating that tribulusamide D could be developed as a potential therapeutic agent for the treatment of inflammatory disorders.

  16. Chlorogenic Acid Protects against Atherosclerosis in ApoE−/− Mice and Promotes Cholesterol Efflux from RAW264.7 Macrophages

    Science.gov (United States)

    Wu, Chongming; Luan, Hong; Zhang, Xue; Wang, Shuai; Zhang, Xiaopo; Sun, Xiaobo; Guo, Peng

    2014-01-01

    Chlorogenic acid (CGA) is one of the most abundant polyphenols in the human diet and is suggested to be a potential antiatherosclerotic agent due to its proposed hypolipidemic, anti-inflammatory and antioxidative properties. The aim of this study was to evaluate the effect of CGA on atherosclerosis development in ApoE−/− mice and its potential mechanism. ApoE−/− mice were fed a cholesterol-rich diet without (control) or with CGA (200 and 400 mg/kg) or atorvastatin (4 mg/kg) for 12 weeks. During the study plasma lipid and inflammatory parameters were determined. Treatment with CGA (400 mg/kg) reduced atherosclerotic lesion area and vascular dilatation in the aortic root, comparable to atorvastatin. CGA (400 mg/kg) also significantly decreased plasma levels of total cholesterol, triglycerides and low-density lipoprotein-cholesterol as well as inflammatory markers. Supplementation with CGA or CGA metabolites-containing serum suppressed oxidized low-density lipoprotein (oxLDL)-induced lipid accumulation and stimulated cholesterol efflux from RAW264.7 cells. CGA significantly increased the mRNA levels of PPARγ, LXRα, ABCA1 and ABCG1 as well as the transcriptional activity of PPARγ. Cholesterol efflux assay showed that three major metabolites, caffeic, ferulic and gallic acids, significantly stimulated cholesterol efflux from RAW264.7 cells. These results suggest that CGA potently reduces atherosclerosis development in ApoE−/− mice and promotes cholesterol efflux from RAW264.7 macrophages. Caffeic, ferulic and gallic acids may be the potential active compounds accounting for the in vivo effect of CGA. PMID:25187964

  17. Ginseng marc-derived low-molecular weight oligosaccharide inhibits the growth of skin melanoma cells via activation of RAW264.7 cells.

    Science.gov (United States)

    Seo, Jeong Yeon; Lee, Chang Won; Choi, Doo Jin; Lee, Jisun; Lee, Jae Yeon; Park, Yong Il

    2015-12-01

    Panax ginseng C.A. Meyer has been traditionally consumed to prevent or treat various medical disorders due to its diverse health benefits. Polysaccharides isolated from Panax ginseng have been known to possess various pharmacological activities, including immune modulating, anti-diabetic, and anti-obesity properties. Despite the increasing number of reports on the bioactivities of ginseng polysaccharides, little is known regarding the medicinal potential of ginseng-derived oligosaccharides. In this study, we prepared a lower-molecular weight oligosaccharide (GOS, MW. 2.2kDa) from ginseng polysaccharides (MW. 11-605kDa) by enzymatic degradation and evaluated for its immunostimulating activities in RAW 264.7 murine macrophage cells. GOS was shown to be a glucan type oligosaccharide mainly containing glucose residues (97.48 in molar %). Treatment with GOS (100-500μg/ml) dose-dependently enhanced the production of TNF-α, IL-6, and NO in RAW 264.7 cells. Western blot analysis indicated that GOS dose-dependently induced the phosphorylation of c-Jun N-terminal kinase (JNK), extracellular signal-regulated kinase (ERK), p38, and nuclear factor κB (NFκB), which are upstream signalling molecules for cytokine production. While GOS was not cytotoxic to the RAW 264.7 macrophage cells at the concentration tested (up to 1000μg/ml), when B16F10 melanoma cells were co-cultured with the GOS-activated macrophages, the cell viability of melanoma cells was dose-dependently decreased through the induction of apoptotic cell death. Taken together, these results suggested that ginseng marc-derived GOS has anti-cancer activity in vitro against melanoma cells by potentiating macrophage function. Copyright © 2015 Elsevier B.V. All rights reserved.

  18. Scavenger Receptor SREC-I Mediated Entry of TLR4 into Lipid Microdomains and Triggered Inflammatory Cytokine Release in RAW 264.7 Cells upon LPS Activation

    Science.gov (United States)

    Murshid, Ayesha; Gong, Jianlin; Prince, Thomas; Borges, Thiago J.; Calderwood, Stuart K.

    2015-01-01

    Scavenger receptor associated with endothelial cells I (SREC-I) was shown to be expressed in immune cells and to play a role in the endocytosis of peptides and antigen presentation. As our previous studies indicated that SREC-I required intact Toll-like receptor 4 (TLR4) expression for its functions in tumor immunity, we examined potential interactions between these two receptors. We have shown here that SREC-I became associated with TLR4 on binding bacterial lipopolysaccharides (LPS) in RAW 264.7 and HEK 293 cells overexpressing these two receptors. The receptors then became internalized together in intracellular endosomes. SREC-I promoted TLR4-induced signal transduction through the NF-kB and MAP kinase pathways, leading to enhanced inflammatory cytokine release. Activation of inflammatory signaling through SREC-I/TLR4 complexes appeared to involve recruitment of the receptors into detergent-insoluble, cholesterol-rich lipid microdomains that contained the small GTPase Cdc42 and the non-receptor tyrosine kinase c-src. Under conditions of SREC-I activation by LPS, TLR4 activity required Cdc42 as well as cholesterol and actin polymerization for signaling through NF-kB and MAP kinase pathways in RAW 264.7 cells. SREC-I appeared to respond differently to another ligand, the molecular chaperone Hsp90 that, while triggering SREC-I-TLR4 binding caused only faint activation of the NF-kB pathway. Our experiments therefore indicated that SREC-I could bind LPS and might be involved in innate inflammatory immune responses to extracellular danger signals in RAW 264.7 cells or bone marrow-derived macrophages. PMID:25836976

  19. Mycotoxin zearalenone induces AIF- and ROS-mediated cell death through p53- and MAPK-dependent signaling pathways in RAW264.7 macrophages.

    Science.gov (United States)

    Yu, Ji-Yeon; Zheng, Zhong-Hua; Son, Young-Ok; Shi, Xianglin; Jang, Young-Oh; Lee, Jeong-Chae

    2011-12-01

    Zearalenone (ZEN) is commonly found in many food commodities and is known to cause reproductive disorders and genotoxic effects. However, the mode of ZEN-induced cell death of macrophages and the mechanisms by which ZEN causes cytotoxicity remain unclear. The present study shows that ZEN treatment reduces viability of RAW264.7 cells in a dose-dependent manner. ZEN causes predominantly necrotic and late apoptotic cell death. ZEN treatment also results in the loss of mitochondrial membrane potential (MMP), mitochondrial changes in Bcl-2 and Bax proteins, and cytoplasmic release of cytochrome c and apoptosis-inducing factor (AIF). Pre-treatment of the cells with either z-VAD-fmk or z-IETD-fmk does not attenuate ZEN-mediated cell death, whereas catalase suppresses the ZEN-induced decrease in viability in RAW264.7 cells. Treating the cells with c-Jun N-terminal kinase (JNK), p38 mitogen-activated protein kinase (MAPK), or p53 inhibitor prevented ZEN-mediated changes, such as MMP loss, cellular reactive oxygen species (ROS) increase, and cell death. JNK or p38 MAPK inhibitor inhibited mitochondrial alterations of Bcl-2 and Bax proteins with attendant decreases in cellular ROS levels. Knockdown of AIF via siRNA transfection also diminished ZEN-induced cell death. Further, adenosine triphosphate was markedly depleted in the ZEN-exposed cells. Collectively, these results suggest that ZEN induces cytotoxicity in RAW264.7 cells via AIF- and ROS-mediated signaling, in which the activations of p53 and JNK/p38 play a key role. Copyright © 2011 Elsevier Ltd. All rights reserved.

  20. Ketamine inhibits tumor necrosis factor secretion by RAW264.7 murine macrophages stimulated with antibiotic-exposed strains of community-associated, methicillin-resistant Staphylococcus aureus

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    Aguirre Carlos

    2011-01-01

    Full Text Available Abstract Background Infections caused by community-associated strains of methicillin-resistant Staphylococcus aureus (CA-MRSA are associated with a marked and prolonged host inflammatory response. In a sepsis simulation model, we tested whether the anesthetic ketamine inhibits the macrophage TNF response to antibiotic-exposed CA-MRSA bacteria via its antagonism of N-methyl-D-aspartate (NMDA receptors. RAW264.7 cells were stimulated for 18 hrs with 105 to 107 CFU/mL inocula of either of two prototypical CA-MRSA isolates, USA300 strain LAC and USA400 strain MW2, in the presence of either vancomycin or daptomycin. One hour before bacterial stimulation, ketamine was added with or without MK-801 (dizocilpine, a chemically unrelated non-competitive NMDA receptor antagonist, APV (D-2-amino-5-phosphono-valerate, a competitive NMDA receptor antagonist, NMDA, or combinations of these agents. Supernatants were collected and assayed for TNF concentration by ELISA. Results RAW264.7 cells exposed to either LAC or MW2 in the presence of daptomycin secreted less TNF than in the presence of vancomycin. The addition of ketamine inhibited macrophage TNF secretion after stimulation with either of the CA-MRSA isolates (LAC, MW2 in the presence of either antibiotic. The NMDA inhibitors, MK-801 and APV, also suppressed macrophage TNF secretion after stimulation with either of the antibiotic-exposed CA-MRSA isolates, and the effect was not additive or synergistic with ketamine. The addition of NMDA substrate augmented TNF secretion in response to the CA-MRSA bacteria, and the addition of APV suppressed the effect of NMDA in a dose-dependent fashion. Conclusions Ketamine inhibits TNF secretion by MRSA-stimulated RAW264.7 macrophages and the mechanism likely involves NMDA receptor antagonism. These findings may have therapeutic significance in MRSA sepsis.

  1. Eupatolide inhibits lipopolysaccharide-induced COX-2 and iNOS expression in RAW264.7 cells by inducing proteasomal degradation of TRAF6.

    Science.gov (United States)

    Lee, Jongkyu; Tae, Nara; Lee, Jung Joon; Kim, Taeho; Lee, Jeong-Hyung

    2010-06-25

    Inula britannica is a traditional medicinal plant used to treat bronchitis, digestive disorders, and inflammation in Eastern Asia. Here, we identified eupatolide, a sesquiterpene lactone from I. britannica, as an inhibitor of cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) expression. Eupatolide inhibited the production of nitric oxide (NO) and prostaglandin E(2) (PGE(2)) as well as iNOS and COX-2 protein expression in lipopolysaccharide (LPS)-stimulated RAW264.7 cells. Eupatolide dose-dependently decreased the mRNA levels and the promoter activities of COX-2 and iNOS in LPS-stimulated RAW264.7 cells. Moreover, eupatolide significantly suppressed the LPS-induced expression of nuclear factor-kappa B (NF-kappaB) and activator protein-1 (AP-1) reporter genes. Pretreatment of eupatolide inhibited LPS-induced phosphorylation and degradation of I kappaB alpha, and phosphorylation of RelA/p65 on Ser-536 as well as the activation of mitogen-activated protein kinases (MAPKs) and Akt in LPS-stimulated RAW264.7 cells. Eupatolide induced proteasomal degradation of tumor necrosis factor receptor-associated factor-6 (TRAF6), and subsequently inhibited LPS-induced TRAF6 polyubiquitination. These results suggest that eupatolide blocks LPS-induced COX-2 and iNOS expression at the transcriptional level through inhibiting the signaling pathways such as NF-kappaB and MAPKs via proteasomal degradation of TRAF6. Taken together, eupatolide may be a novel anti-inflammatory agent that induces proteasomal degradation of TRAF6, and a valuable compound for modulating inflammatory conditions. (c) 2010 Elsevier B.V. All rights reserved.

  2. N‑trans‑ρ‑caffeoyl tyramine isolated from Tribulus terrestris exerts anti‑inflammatory effects in lipopolysaccharide‑stimulated RAW 264.7 cells.

    Science.gov (United States)

    Ko, Han-Jik; Ahn, Eun-Kyung; Oh, Joa Sub

    2015-10-01

    Inflammation is induced by the expression of cyclooxygenase‑2 (COX‑2), which is an important mediator of chronic inflammatory diseases, such as rheumatoid arthritis, asthma and inflammatory bowel disease. Tribulus terrestris (T. terrestris) is known to have a beneficial effect on inflammatory diseases. In this study, we investigated the effects of N‑trans‑ρ‑caffeoyl tyramine (CT) isolated from T. terrestris on the production of nitric oxide (NO), and the expression of pro‑inflammatory cytokines and COX‑2 in lipopolysaccharide (LPS)‑stimulated RAW 264.7 cells. We also aimed to elucidate the molecular mechanisms involved. We found that the ethanolic extract of T. terrestris (EETT) and CT inhibited the production of NO, tumor necrosis factor‑α (TNF‑α), interleukin (IL)‑6 and IL‑10 in the LPS‑stimulated RAW 264.7 cells in a dose‑dependent manner. They were determined by reverse transcription-polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA). In addition, CT markedly suppressed the expression of COX‑2 and the production of prostaglandin E2 (PGE2) in response to LPS stimulation. Furthermore, CT markedly decreased p‑c‑Jun N‑terminal kinase (p‑JNK) protein expression in LPS‑stimulated RAW 264.7 cells. COX-2 and p-JNK were measured by western blot analysis. Taken together, these findings indicate that CT isolated from T. terrestris is a novel and potent modulator of inflammatory responses. Thus, it may prove benefiical to further evaluate CT as a possible treatment for chronic inflammatory diseases.

  3. Suppressive Effect on Lipopolysaccharide-Induced Proinflammatory Mediators by Citrus aurantium L. in Macrophage RAW 264.7 Cells via NF-κB Signal Pathway

    Directory of Open Access Journals (Sweden)

    Sang-Rim Kang

    2011-01-01

    Full Text Available Citrus fruits have been used as an edible fruit and a traditional medicine since ancient times. In particular, the peels of immature citrus fruits are used widely in traditional herbal medicine in Korea, as they are believed to contain bioactive components exerting anti-inflammatory activity. This study examined whether the crude methanol extract of Citrus aurantium L. (CME has a suppressive effect on inducible enzymes and proinflammatory cytokines by inhibiting the NF-κB pathway in LPS-stimulated macrophage RAW 264.7 cells. The cells were pretreated with the indicated concentrations of CME (5, 10, 20, and 50 μg/mL and then treated with LPS (1 μg/mL. The results showed that CME (10, 20, and 50 μg/mL inhibited the LPS- (1 μg/mL induced mRNA and protein expression of iNOS in macrophage Raw 264.7 cells. In addition, the expression of COX-2 was inhibited at the mRNA and protein levels by CME in a dose-dependent manner. The mRNA expression of proinflammatory cytokines, such as TNF-α and IL-6, were markedly reduced by CME (10, 20, and 50 μg/mL. Moreover, CME clearly suppressed the nuclear translocation of the NF-κB p65 subunits, which was correlated with its inhibitory effect on I-κB phosphorylation. These results suggest that CME has anti-inflammatory properties by modulating the expression of COX-2, iNOS, and proinflammatory cytokines, such as TNF-α and IL-6, in macrophage RAW 264.7 cells via the NF-κB pathway.

  4. Anti-Inflammatory Activity of Heterocarpin from the Salt Marsh Plant Corydalis heterocarpa in LPS-Induced RAW 264.7 Macrophage Cells

    Directory of Open Access Journals (Sweden)

    You Ah Kim

    2015-08-01

    Full Text Available The inhibitory effect of three chromones 1–3 and two coumarins 4–5 on the production of nitric oxide (NO was evaluated in LPS-induced RAW 264.7 macrophage cells. Among the compounds tested heterocarpin (1, a furochromone, significantly inhibited its production in a dose-dependent manner. In addition, heterocarpin suppressed prostaglandin E2 (PGE2 production and expression of cytokines such as inducible nitric oxide synthase (iNOS, cyclooxygenase-2 (COX-2, tumor necrosis factor-α (TNF-α, interleukin-1β (IL-1β and interleukin-6 (IL-6.

  5. Immunomodulatory Efficacy of Standardized Annona muricata (Graviola) Leaf Extract via Activation of Mitogen-Activated Protein Kinase Pathways in RAW 264.7 Macrophages

    OpenAIRE

    Kim, Goon-Tae; Tran, Nguyen Khoi Song; Choi, Eun-Hye; Song, Yoo-Jeong; Song, Jae-Hwi; Shim, Soon-Mi; Park, Tae-Sik

    2016-01-01

    Annona muricata, commonly known as Graviola, has been utilized as a traditional medicine to treat various human diseases. The aim of this study was to examine the immune-enhancing activity of Graviola leaf extracts in RAW 264.7 macrophage cells. Active ingredients in Graviola leaf extracts (GE) were identified as kaempferol-3-O-rutinoside and quercetin-3-O-rutinoside by LC-MS/MS. When treated with steam or 50% ethanol GE, cell morphology was altered due to initiation of cell differentiation. ...

  6. A High-Throughput Hardware Architecture for the H.264/AVC Half-Pixel Motion Estimation Targeting High-Definition Videos

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    Marcel M. Corrêa

    2011-01-01

    Full Text Available This paper presents a high-performance hardware architecture for the H.264/AVC Half-Pixel Motion Estimation that targets high-definition videos. This design can process very high-definition videos like QHDTV (3840×2048 in real time (30 frames per second. It also presents an optimized arrangement of interpolated samples, which is the main key to achieve an efficient search. The interpolation process is interleaved with the SAD calculation and comparison, allowing the high throughput. The architecture was fully described in VHDL, synthesized for two different Xilinx FPGA devices, and it achieved very good results when compared to related works.

  7. Echinacea increases arginase activity and has anti-inflammatory properties in RAW 264.7 macrophage cells, indicative of alternative macrophage activation.

    Science.gov (United States)

    Zhai, Zili; Solco, Avery; Wu, Lankun; Wurtele, Eve S; Kohut, Marian L; Murphy, Patricia A; Cunnick, Joan E

    2009-02-25

    The genus Echinacea is a popular herbal immunomodulator. Recent reports indicate that Echinacea products inhibit nitric oxide (NO) production in activated macrophages. In the present study we determined the inhibitory effects of alcohol extracts and individual fractions of alcohol extracts of Echinacea on NO production, and explored the mechanism underlying the pharmacological anti-inflammatory activity. Alcohol extracts of three medicinal Echinacea species, Echinacea angustifolia, Echinacea pallida and Echinacea purpurea, were prepared using Soxhlet apparatus and fractionated using HPLC. NO production by LPS activated RAW 264.7 macrophage cells was measured using a Griess reagent and iNOS detected using immunoblotting. In addition, effects on arginase activity were measured in RAW 264.7 cells stimulated with 8-bromo-cAMP +/- LPS. Alcohol extracts of all three Echinacea species significantly inhibited NO production by lipopolysaccharide (LPS)-activated the RAW 264.7 macrophage cell line; among them Echinacea pallida was the most active. The Echinacea-mediated decrease in NO production was unlikely due to a direct scavenging of NO because the extracts did not directly inhibit NO released from an NO donor, sodium nitroprusside. An immunoblotting assay demonstrated that the extract of Echinacea pallida inhibited inducible nitric oxide synthase (iNOS) protein expression in LPS-treated macrophages. The enzymes iNOS and arginase metabolize a common substrate, l-arginine, but produce distinct biological effects. While iNOS is involved in inflammatory response and host defense, arginase participates actively in anti-inflammatory activation. Arginase activity of RAW 264.7 cells stimulated with 8-bromo-cAMP was significantly increased by alcohol extracts of all three Echinacea species. The polar fraction containing caffeic acid derivatives enhanced arginase activity, while the lipophilic fraction containing alkamides exhibited a potential of inhibiting NO production and i

  8. Emu Oil Reduces LPS-Induced Production of Nitric Oxide and TNF-α but not Phagocytosis in RAW 264 Macrophages.

    Science.gov (United States)

    Miyashita, Tadayoshi; Minami, Kazuhiro; Ito, Minoru; Koizumi, Ryosuke; Sagane, Yoshimasa; Watanabe, Toshihiro; Niwa, Koichi

    2018-03-09

    Emu is the second-largest extant bird native to Australia. Emu oil, obtained from the emu's fat deposits, is used as an ingredient in cosmetic skincare products. Emu oil has been reported to improve several inflammatory symptoms; however, the mechanisms of these anti-inflammatory effects are largely unknown. This study investigated the effects of emu oil on the inflammatory macrophage response in vitro. A murine macrophage cell line, RAW 264, was incubated in culture media supplemented with or without emu oil and stimulated with lipopolysaccharide (LPS). We determined phagocytic activity by measuring the number of fluorescent microspheres taken up by the cells. The phagocytic activity of RAW 264 cells in the presence of LPS was unaffected by emu oil. We also determined production of nitric oxide (NO) and tumor necrosis factor (TNF)-α in the culture medium using the Griess reaction and an enzyme-linked immunosorbent assay, respectively, and the protein expression of inducible NO synthase (iNOS) using western blotting. The results indicated that emu oil reduced the LPS-induced production of NO, TNF-α, and iNOS expression in a dose-dependent manner. The results suggested that emu oil does not reduce the phagocytic clearance rate of inflammatory matter; however, it does reduce the production of NO and TNFα in macrophages. These latter products enhance the inflammatory response and emu oil thereby demonstrated anti-inflammatory properties.

  9. Control of microorganisms of oral health interest with Arctium lappa L. (burdock) extract non-cytotoxic to cell culture of macrophages (RAW 264.7).

    Science.gov (United States)

    de Oliveira, Jonatas Rafael; de Aguiar Almeida, Rosilene Batista; das Graças Figueiredo Vilela, Polyana; de Oliveira, Felipe Eduardo; da Rocha, Rosilene Fernandes; Jorge, Antonio Olavo Cardoso; de Oliveira, Luciane Dias

    2014-08-01

    To evaluate the antimicrobial activity of Arctium lappa L. extract on Staphylococcus aureus, S. epidermidis, Streptococcus mutans, Candida albicans, C. tropicalis and C. glabrata. In addition, the cytotoxicity of this extract was analyzed on macrophages (RAW 264.7). By broth microdilution method, different concentrations of the extract (250-0.4 mg/mL) were used in order to determine the minimum microbicidal concentration (MMC) in planktonic cultures and the most effective concentration was used on biofilms on discs made of acrylic resin. The cytotoxicity A. lappa L. extract MMC was evaluated on RAW 264.7 by MTT assay and the quantification of IL-1β and TNF-α by ELISA. The most effective concentration was 250 mg/mL and also promoted significant reduction (log₁₀) in the biofilms of S. aureus (0.438 ± 0.269), S. epidermidis (0.377 ± 0.298), S. mutans (0.244 ± 0.161) and C. albicans (0.746 ± 0.209). Cell viability was similar to 100%. The production of IL-1β was similar to the control group (p>0.05) and there was inhibition of TNF-α (plappa L. extract was microbicidal for all the evaluated strains in planktonic cultures, microbiostatic for biofilms and not cytotoxic to the macrophages. Copyright © 2014 Elsevier Ltd. All rights reserved.

  10. New ent-kauranes from the fruits of Annona glabra and their inhibitory nitric oxide production in LPS-stimulated RAW264.7 macrophages.

    Science.gov (United States)

    Nhiem, Nguyen Xuan; Hien, Nguyen Thi Thu; Tai, Bui Huu; Anh, Hoang Le Tuan; Hang, Dan Thi Thuy; Quang, Tran Hong; Kiem, Phan Van; Minh, Chau Van; Ko, Wonmin; Lee, Seungjun; Oh, Hyuncheol; Kim, Seung Hyun; Kim, Young Ho

    2015-01-15

    Three new ent-kaurane diterpenoids, 7β,16α,17-trihydroxy-ent-kauran-19-oic acid (1), 7β,17-dihydroxy-16α-ent-kauran-19-oic acid 19-O-β-d-glucopyranoside ester (2), 7β,17-dihydroxy-ent-kaur-15-en-19-oic acid 19-O-β-d-glucopyranoside ester (3) along with five known compounds, paniculoside IV (4), 16α,17-dihydroxy-ent-kaurane (5), 16β,17-dihydroxy-ent-kaurane (6), 16β,17-dihydroxy-ent-kauran-19-al (7), and 16β,17-dihydroxy-ent-kauran-19-oic acid (8) were isolated from the fruits of Annona glabra. Their chemical structures were elucidated by physical and chemical methods. All compounds were evaluated for inhibitory activity against nitric oxide (NO) production in LPS-stimulated RAW 264.7 macrophages. As the results, compound 3 showed potent inhibitory LPS-stimulated NO production in RAW 264.7 macrophages with the IC50 value of 0.01±0.01μM; compounds 1 and 7 showed significant inhibitory NO production with the IC50 values of 0.39±0.12μM and 0.32±0.04μM, respectively. Copyright © 2014 Elsevier Ltd. All rights reserved.

  11. Anti-inflammatory effects of methanol extracts of the root of Lilium lancifolium on LPS-stimulated Raw264.7 cells.

    Science.gov (United States)

    Kwon, Ok-Kyoung; Lee, Mee-Young; Yuk, Ji-Eun; Oh, Sei-Ryang; Chin, Young-Won; Lee, Hyeong-Kyu; Ahn, Kyung-Seop

    2010-07-06

    Lilium lancifolium is commonly used to treat bronchitis, pneumonia, etc. In this study, we investigated the anti-inflammatory effects of methanol extracts of the root of Lilium lancifolium (LL extracts) in LPS-stimulated Raw264.7 cells. Levels of NO, PGE(2) and pro-inflammatory cytokines (IL-6 and TNF-alpha) in the supernatant fraction were determined using sandwich ELISA. Expression of COX-2 and iNOS, phosphorylation of MAPK subgroups (ERK and JNK), and NF-kappaB activation in extracts were detected via Western blot and immunocytochemistry assays. The LL extract significantly inhibited NO, PGE(2), IL-6 and TNF-alpha production in LPS-stimulated cells, and suppressed iNOS and COX-2 expression. A mechanism-based study showed that phosphorylation of ERK1/2 and JNK and translocation of the NF-kappaB p65 subunit into nuclei were inhibited by the LL extract. Furthermore, interleukin-4 and interleukin-13 production in Con A-induced splenocytes was suppressed. These results indicate that anti-inflammatory effects of methanol extracts from Lilium lancifolium are due to downregulation of iNOS and COX-2 via suppression of NF-kappaB activation and nuclear translocation as well as blocking of ERK and JNK signaling in LPS-stimulated Raw264.7 cells. Copyright (c) 2010 Elsevier Ireland Ltd. All rights reserved.

  12. PEP-1-PON1 protein regulates inflammatory response in raw 264.7 macrophages and ameliorates inflammation in a TPA-induced animal model.

    Directory of Open Access Journals (Sweden)

    Mi Jin Kim

    Full Text Available Paraoxonase 1 (PON1 is an antioxidant enzyme which plays a central role in various diseases. However, the mechanism and function of PON1 protein in inflammation are poorly understood. Since PON1 protein alone cannot be delivered into cells, we generated a cell permeable PEP-1-PON1 protein using protein transduction domains, and examined whether it can protect against cell death in lipopolysaccharide (LPS or hydrogen peroxide (H2O2-treated Raw 264.7 cells as well as mice with 12-O-tetradecanoyl phorbol-13-acetate (TPA-induced skin inflammation. We demonstrated that PEP-1-PON1 protein transduced into Raw 264.7 cells and markedly protected against LPS or H2O2-induced cell death by inhibiting cellular reactive oxygen species (ROS levels, the inflammatory mediator's expression, activation of mitogen-activated protein kinases (MAPKs and cellular apoptosis. Furthermore, topically applied PEP-1-PON1 protein ameliorates TPA-treated mice skin inflammation via a reduction of inflammatory response. Our results indicate that PEP-1-PON1 protein plays a key role in inflammation and oxidative stress in vitro and in vivo. Therefore, we suggest that PEP-1-PON1 protein may provide a potential protein therapy against oxidative stress and inflammation.

  13. Preparation of procyanidin B2 from apple pomace and its inhibitory effect on the expression of cyclooxygenase-2 in lipopolysaccharide-treated RAW264.7 macrophages

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    Huawei Zhang

    2011-06-01

    Full Text Available Dimeric procyanidin B2 (PB2 is one of phenolic compounds in apple pomace, an agro-industrial byproduct in apple juice processing. This work focused on purification of PB2 from apple pomace using sephadex column chromatography and its potential effect on lipopolysaccharide (LPS-induced inflammation using RAW264.7 macrophages. PB2 with the purity of 72.28 ± 1.85% was successfully afforded using resin and gel column chromatographic technique. Anti-inflammatory tests suggested that the expression of cyclooxygenase-2 (COX-2 in LPS-induced murine RAW264.7 macrophages was suppressed in a PB2 concentration-dependent manner. PB2 at no less than 50 μg·mL-1 could significantly suppress inflammation in the LPS-induced cells. Moreover, this suppressive effect was not correlated with PB2 pretreating. However, the COX-2 expression was not reduced in LPS pretreatment way followed by PB2 exposure, which suggested that PB2 has no repairing function. The results showed that high pure PB2 prepared from apple pomace has a remarkable anti-inflammatory property.

  14. Suppression of lipopolysaccharide-induced of inducible nitric oxide synthase and cyclooxygenase-2 by Sanguis Draconis, a dragon's blood resin, in RAW 264.7 cells.

    Science.gov (United States)

    Choy, Cheuk-Sing; Hu, Chien-Ming; Chiu, Wen-Ta; Lam, Carlos-Shu Kei; Ting, Yih; Tsai, Shin-Han; Wang, Tzu-Chien

    2008-02-12

    Sanguis Draconis (SD) is a kind of dragon's blood resin that is obtained from Daemomorops draco (Palmae). It is used in traditional medicine and has shown anti-inflammatory activity in some diseases. In this study, we examined the effects of Sanguis Dranonis ethanol extract (SDEE) on LPS-induced inflammation using RAW 264.7 cells. Our data indicated that SDEE inhibits LPS-stimulated NO, PGE2, IL-1 beta and TNF-alpha release, and iNOS and COX-2 expression. Furthermore, SDEE suppressed the LPS-induced p65 expression of NF-kappa B, which was associated with the inhibition of I kappa B-alpha degradation. We also found that the expression of HO-1 was significantly increased in RAW 264.7 cells by SDEE. These results suggest among possibilities of anti-inflammation that SDEE inhibits the production of NO and PGE2 by the down-regulation of iNOS and COX-2 gene expression via the suppression of NF-kappaB (p65) activation. SDEE can induce HO-1 over-expression in macrophage cells, which indicates that it may possess antioxidant properties. This result means that SEDD its anti-inflammatory effects in macrophages may be through a novel mechanism that involves the action of HO-1. Thus, SD could provide a potential therapeutic approach for inflammation-associated disorders.

  15. An efficient HW and SW design of H.264 video compression, storage and playback on FPGA devices for handheld thermal imaging systems

    Science.gov (United States)

    Gunay, Omer; Ozsarac, Ismail; Kamisli, Fatih

    2017-05-01

    Video recording is an essential property of new generation military imaging systems. Playback of the stored video on the same device is also desirable as it provides several operational benefits to end users. Two very important constraints for many military imaging systems, especially for hand-held devices and thermal weapon sights, are power consumption and size. To meet these constraints, it is essential to perform most of the processing applied to the video signal, such as preprocessing, compression, storing, decoding, playback and other system functions on a single programmable chip, such as FPGA, DSP, GPU or ASIC. In this work, H.264/AVC (Advanced Video Coding) compatible video compression, storage, decoding and playback blocks are efficiently designed and implemented on FPGA platforms using FPGA fabric and Altera NIOS II soft processor. Many subblocks that are used in video encoding are also used during video decoding in order to save FPGA resources and power. Computationally complex blocks are designed using FPGA fabric, while blocks such as SD card write/read, H.264 syntax decoding and CAVLC decoding are done using NIOS processor to benefit from software flexibility. In addition, to keep power consumption low, the system was designed to require limited external memory access. The design was tested using 640x480 25 fps thermal camera on CYCLONE V FPGA, which is the ALTERA's lowest power FPGA family, and consumes lower than 40% of CYCLONE V 5CEFA7 FPGA resources on average.

  16. Arctigenin inhibits lipopolysaccharide-induced iNOS expression in RAW264.7 cells through suppressing JAK-STAT signal pathway.

    Science.gov (United States)

    Kou, Xianjuan; Qi, Shimei; Dai, Wuxing; Luo, Lan; Yin, Zhimin

    2011-08-01

    Arctigenin has been demonstrated to have an anti-inflammatory function, but the precise mechanisms of its action remain to be fully defined. In the present study, we determined the effects of arctigenin on lipopolysaccharide (LPS)-induced production of proinflammatory mediators and the underlying mechanisms involved in RAW264.7 cells. Our results indicated that arctigenin exerted its anti-inflammatory effect by inhibiting ROS-dependent STAT signaling through its antioxidant activity. Arctigenin also significantly reduced the phosphorylation of STAT1 and STAT 3 as well as JAK2 in LPS-stimulated RAW264.7 cells. The inhibitions of STAT1 and STAT 3 by arctigenin prevented their translocation to the nucleus and consequently inhibited expression of iNOS, thereby suppressing the expression of inflammation-associated genes, such as IL-1β, IL-6 and MCP-1, whose promoters contain STAT-binding elements. However, COX-2 expression was slightly inhibited at higher drug concentrations (50 μM). Our data demonstrate that arctigenin inhibits iNOS expression via suppressing JAK-STAT signaling pathway in macrophages. Crown Copyright © 2011. Published by Elsevier B.V. All rights reserved.

  17. Ability of certain plant extracts traditionally used to treat ciguatera fish poisoning to inhibit nitric oxide production in RAW 264.7 macrophages.

    Science.gov (United States)

    Kumar-Roiné, Shilpa; Matsui, Mariko; Reybier, Karine; Darius, Hélène Taiana; Chinain, Mireille; Pauillac, Serge; Laurent, Dominique

    2009-06-25

    Ciguatera fish poisoning (CFP) is an intertropical ichthyosarcotoxism that manifests in complex assortment of symptoms in humans. Ciguatoxins (CTXs), issued from Gambierdicus spp., are causative agents of this intoxication. We have recently demonstrated that a Pacific CTX (P-CTX-1B) strongly modulated iNOS expression, leading to overproduction of nitric oxide (NO) in RAW 264.7 murine macrophage cells. NO produced in large amounts is involved in a wide range of pathophysiological processes. Many traditional remedies are commonly used in the Pacific against CFP. In this context, bioassay-guided screening was carried out to study NO inhibiting capacity of 28 selected plant extracts. We prepared aqueous extracts of plants used in New Caledonia in the treatment of CFP and screened their NO inhibitory activity in lipopolysaccharide (LPS)-activated RAW 264.7 macrophages. Among 28 plants tested, Euphorbia hirta (Euphorbiaceae), Syzygium malaccense (Myrtaceae), Schinus terebenthifolius (Anacardiaceae), Punica granatum (Punicaceae), Cerbera manghas (Apocynaceae), Vitex trifolia (Labiateae) and Ximenia americana (Olacaceae) showed inhibitory activity, validating their use as traditional remedies in CFP, and the potential for use in the treatment of conditions accompanied by NO overproduction. These plants are promising candidates for further screening of their active compounds through activity-guided fractionation.

  18. Anti-inflammatory effects of chloroform soluble fraction from Perilla frutescens britton leaves produced by radiation breeding in RAW264.7 cells

    Energy Technology Data Exchange (ETDEWEB)

    Jo, Yun Ho; So, Yang Kang; Kim, Jin Baek; Jin, Chang Hyun [Advance Radiation Technology Institute, Korea Atomic Energy Research Institute, Jeongeup (Korea, Republic of); Lee, Jun Soo [Dept. Food Science and Technology Graduate School, Chungbuk National University, Cheongju (Korea, Republic of); Lee, Seung Young [Freshwater Bioresources Utilization Division, Nakdonggang National Institute of Biological Resources, Sangju (Korea, Republic of)

    2016-11-15

    The present study aimed to determine the anti-inflammatory effects of each solvent fraction of a mutant Perilla frutescens produced by radiation breeding. Following extraction with 80% methanol, P. frutescens was fractionated in the order of hexane, chloroform, ethyl acetate, and butanol; the chloroform fraction exhibited less cytotoxicity, the greatest inhibitory effect on the production of nitric oxide (NO), and the highest rate of inhibition on the generation of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), monocyte chemoattractant protein-1 (MCP-1), and interferon-β (IFN-β). The chloroform fraction also suppressed the mRNA and protein levels of inducible nitric oxide synthase (iNOS) and reduced the activation of nuclear factor-{sub κ}B (NF-{sub κ}B) in lipopolysaccharide (LPS)-stimulated RAW264.7 cells. Finally, the presence of corosolic acid in the chloroform fraction was identifed. Taken together, the present fndings indicate that the chloroform fraction obtained from mutant P. frutescens inhibited NO production in LPSstimulated RAW264.7 cells via the suppression of iNOS expression and the inactivation of NF-{sub κ}B.

  19. Terminalia chebula Fructus Inhibits Migration and Proliferation of Vascular Smooth Muscle Cells and Production of Inflammatory Mediators in RAW 264.7

    Directory of Open Access Journals (Sweden)

    Hyun-Ho Lee

    2015-01-01

    Full Text Available Pathogenesis of atherosclerosis and neointima formation after angioplasty involves vascular smooth muscle cells (VSMCs migration and proliferation followed by inflammatory responses mediated by recruited macrophages in the neointima. Terminalia chebula is widely used traditional medicine in Asia for its beneficial effects against cancer, diabetes, and bacterial infection. The study was designed to determine whether Terminalia chebula fructus water extract (TFW suppresses VSMC migration and proliferation and inflammatory mediators production in macrophage (RAW 264.7. Our results showed that TFW possessed strong antioxidative effects in 1,1-diphenyl-2-picryl hydrazyl (DPPH scavenging and lipid peroxidation assays. In addition, TFW reduced nitric oxide (NO production, inducible nitric oxide synthase (iNOS, and cyclooxygenase-2 (COX-2 expression in RAW 264.7 cells. Also, TFW inhibited platelet-derived growth factor (PDGF-BB induced VSMC migration as determined by wound healing and Boyden chamber assays. The antimigratory effect of TFW was due to its inhibitory effect on metalloproteinase-9 (MMP-9 expression, focal adhesion kinase (FAK activation, and Rho-family of small GTPases (Cdc42 and RhoA expression in VSMCs. Furthermore, TFW suppressed PDGF-BB induced VSMC proliferation by downregulation of mitogen activated protein kinases (MAPKs signaling molecules. These results suggest that TFW could be a beneficial resource in the prevention of atherosclerosis.

  20. Falcarindiol inhibits LPS-induced inflammation via attenuating MAPK and JAK-STAT signaling pathways in murine macrophage RAW 264.7 cells.

    Science.gov (United States)

    Venkatesan, Thamizhiniyan; Choi, Young-Woong; Lee, Jennifer; Kim, Young-Kyoon

    2018-01-24

    Falcarindiol (FAD) is a natural polyacetylene compound found rich in many plants of the Umbelliferae family. Previously, we isolated FAD from the rhizome of Cnidium officinale Makino, which belongs to the Umbelliferae family and found it to have a significant inhibitory effect on lipopolysaccharide (LPS)-induced production of nitric oxide, a pro-inflammatory molecule in murine macrophage RAW 264.7 cells. In this study, we investigated its effect on the expression of other major pro-inflammatory molecules as well as the mechanism underlying these effects. Pre-treatment of RAW 264.7 cells with FAD suppressed LPS-stimulated mRNA expression of inducible nitric oxide synthase (iNOS), tumor necrosis factor alpha (TNFα), interleukin-6 (IL-6), and interleukin-1 beta (IL-1β) and thereby reduced the respective protein levels. Mechanistic studies demonstrated that FAD attenuated the LPS-induced activation of JNK, ERK, STAT1, and STAT3 signaling molecules. Moreover, we found that FAD did not influence LPS-induced activation of p38 and NFκB signaling pathways. Collectively, this study provides evidence that FAD inhibits the production of major pro-inflammatory molecules in LPS-challenged murine macrophages via suppression of JNK, ERK, and STAT signaling pathways.

  1. Effects of TiO2 nanotube layers on RAW 264.7 macrophage behaviour and bone morphogenetic protein-2 expression.

    Science.gov (United States)

    Sun, S J; Yu, W Q; Zhang, Y L; Jiang, X Q; Zhang, F Q

    2013-12-01

    To investigate behaviour and osteogenic cytokine expression of RAW264.7 macrophages grown on TiO2 nanotube layers. The murine macrophage cell line RAW 264.7 was cultured on TiO2 nanotubes of varying diameter; macrophage morphology was examined using scanning electron microscopy. Cell adhesion and viability were assessed with the aid of the MTT method and BMP-2 and TGF-β gene expression were examined by RT-PCR analysis. Levels of BMP-2, TGF-β1 and ICAM-1 proteins secreted into the supernatant were measured by ELISA assay. Macrophages cultured on nanotube layers had spread out morphology, the largest (120 nm) nanotube layer eliciting an elongation by 24 h. Macrophages adhered significantly less to 120 nm TiO2 nanotubes than to control discs at 4 h after application; after 24 h incubation, macrophages were sufficiently viable (P nanotube layers. Increasing nanotube diameter led to increased BMP-2 protein secretion and increased BMP-2 mRNA expression. These results demonstrate that nanoscale topography of TiO2 nanotube layers can affect macrophage morphology, adhesion, viability and BMP-2 expression. Macrophages grown on layers of large nanotubes had the highest potential to enhance bone formation during bone healing. © 2013 John Wiley & Sons Ltd.

  2. Responses of RAW264.7 macrophages to water-dispersible gold and silver nanoparticles stabilized by metal-carbon σ-bonds.

    Science.gov (United States)

    Hashimoto, Masanori; Toshima, Hirokazu; Yonezawa, Tetsu; Kawai, Koji; Narushima, Takashi; Kaga, Masayuki; Endo, Kazuhiko

    2014-06-01

    Nanometals are currently receiving considerable attention for industrial and biomedical applications, but their potentially hazardous and toxic effects have not been extensively studied. This study evaluated the biological responses of novel water-dispersible gold (Au-NPs) and silver nanoparticles (Ag-NPs) stabilized by Au-C or Ag-C σ-bonds in cultured macrophages (RAW264.7), via analysis of the cell viability, the integrity of the plasma membrane, and the inflammatory and morphological properties. The cultured RAW264.7 was exposed to metal-NPs at various concentrations. The Ag-NPs showed cytotoxicity at high NP concentrations, but the cytotoxic effects of the Au-NPs were smaller than those of the Ag-NPs. For the microscopic analysis, both types of particles were internalized into cells, the morphological changes in the cells which manifested as an expansion of the vesicles' volume, were smaller for the Au-NPs compared with the Ag-NPs. For the Ag-NPs, the endocytosis abilities of the macrophages might have induced harmful effects, because of the expansion of the cell vesicles. Although an inflammatory response was observed for both the Au- and Ag-NPs, the harmful effects of the Au-NPs were smaller than those of the Ag-NPs, with minor morphological changes observed even after internalization of the NPs into the cells. Copyright © 2013 Society of Plastics Engineers.

  3. Anti-inflammatory effects of β-sitosterol-β-D-glucoside from Trachelospermum jasminoides (Apocynaceae) in lipopolysaccharide-stimulated RAW 264.7 murine macrophages.

    Science.gov (United States)

    Choi, Ji Na; Choi, Yun-Hyeok; Lee, Jeong-Min; Noh, Ill Chan; Park, Jae Won; Choi, Wahn Soo; Choi, Jung Ho

    2012-01-01

    Trachelospermum jasminoides (Apocynaceae) has pharmacological effects that include anti-inflammatory, anti-bacterial and anti-viral activities, which have been observed from various studies. Of these pharmacological effects, the anti-inflammatory capacity of compounds from T. jasminoides is not yet known exactly. In this study, we investigated the compound that can be used for the suppression of lipopolysacchaide (LPS) stimulated inflammatory responses in macrophages among the five isolated compounds. β-sitosterol-β-D-glucoside (1) was found to reduce nitric oxide (NO) production from LPS-induced RAW 264.7 cells the most. In addition, compound 1 strongly inhibited the interleukin 6 (IL-6) activities of stimulated macrophages. Treatment of RAW 264.7 cells with compound 1 reduced secretion of inflammatory elements including tumour necrosis factor - alpha (TNF-α) and interleukin 1 beta (IL-1β). Thus, compound 1 may be a useful candidate for the development of new drugs to treat endotoxemia and inflammation accompanied by the overproduction of NO.

  4. XH-14, a novel danshen methoxybenzo[b]furan derivative, exhibits anti-inflammatory properties in lipopolysaccharide-treated RAW 264.7 cells

    Directory of Open Access Journals (Sweden)

    Park Geun-Mook

    2013-01-01

    Full Text Available Abstract Background XH-14 isolated from Salvia miltiorrhiza is a bioactive component and adenosine antagonist. In the present study, we evaluated anti-inflammatory properties of XH-14 in murine macrophages. Methods RAW 264.7 murine macrophage cell line was cultured with various concentrations of XH-14 in the absence or presence of lipopolysaccharide (LPS. LPS-induced release and mRNA expression of inflammatory mediators were examined by ELISA and real-time PCR. The modification of signal pathways involved in inflammatory reactions was determined by Western blotting analysis. Results XH-14 suppressed the generation of nitric oxide (NO and prostaglandin E2, and the expression of inducible NO synthase and cyclooxygenase-2 induced by LPS. Similarly, XH-14 inhibited the release of pro-inflammatory cytokines induced by LPS in RAW 264.7 cells. The underlying mechanism of XH-14 on anti-inflammatory action was correlated with down-regulation of mitogen-activated protein kinase and activator protein-1 activation. Conclusions XH-14 inhibits the production of several inflammatory mediators and so might be useful for the treatment of various inflammatory diseases.

  5. Anti-inflammatory activity of saponins from quinoa (Chenopodium quinoa Willd.) seeds in lipopolysaccharide-stimulated RAW 264.7 macrophages cells.

    Science.gov (United States)

    Yao, Yang; Yang, Xiushi; Shi, Zhenxing; Ren, Guixing

    2014-05-01

    Quinoa (Chenopodium quinoa Willd.) is a pseudocereal from South Americas that has received increased interest around the world because it is a good source of different nutrients and rich in saponins. However, the saponins in quinoa seeds planted in China were poorly known. We obtained 4 quinoa saponin fractions, Q30, Q50, Q70, and Q90, and 11 saponins were determined by HPLC-MS. Q50 possessed 8 individual saponins and had the highest content of saponins. We further evaluated the anti-inflammatory activity on RAW 264.7 murine macrophage cells of the 4 fractions. The 4 fractions not only dose-dependently decreased the production of inflammatory mediators NO but also inhibited the release of inflammatory cytokines including tumor necrosis factor-α and interleukin-6 in lipopolysaccharide-induced RAW264.7 cells. These results suggest that quinoa saponins may be used as functional food components for prevention and treatment of inflammation. Our findings demonstrate that saponins from the quinoa have the potential to anti-inflammation by suppressing the release of inflammatory cytokines. © 2014 Institute of Food Technologists®

  6. Allium hookeri root extract exerts anti-inflammatory effects by nuclear factor-κB down-regulation in lipopolysaccharide-induced RAW264.7 cells.

    Science.gov (United States)

    Jang, Ja-Young; Lee, Min-Jung; You, Bo-Ram; Jin, Jong-Sik; Lee, Sung-Hyen; Yun, Ye-Rang; Kim, Hyun Ju

    2017-02-23

    Allium hookeri (AH) is widely consumed as a vegetable and herbal medicine in southeastern Asia. AH has been reported antioxidant, antimicrobial, improvement of bone health and antidiabetic effects. In the present study, we investigated the inhibitory effect of a methanol extract of AH root (AHE) on inflammatory response in lipopolysaccharide (LPS)-induced RAW264.7 cells. Initially, characterization of organic sulfur compounds in AHE was determined using high performance liquid chromatography-electrospray ionization-mass spectrometry (HPLC-ESI-MS). Cells were incubated with LPS and AHE for 24 h. The productions of nitric oxide (NO), reactive oxygen species (ROS), and inflammation-related cytokines were examined. Gene and protein expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) were assessed by polymerase chain reaction and Western blotting. Key factor, nuclear factor kappa B (NF-κB) was also determined. AHE contained organosulfur compounds such as alliin and S-allylcysteine by HPLC-ESI-MS. AHE significantly inhibited NO, ROS, and cytokines production in LPS-induced RAW264.7 cells. In addition, AHE treatment inhibited iNOS and COX-2 mRNA and protein levels, leading to a decrease in iNOS-derived NO level. Furthermore, NF-κB activation was, at least in part, suppressed by AHE treatment. Our data suggest that AHE treatment inhibits the inflammation condition through suppression of iNOS and COX-2 expression via NF-κB down-regulation.

  7. Anti-inflammatory Effects of KOTMIN13: A Mixed Herbal Medicine in LPS-stimulated RAW 264.7 Cells and Mouse Edema Models

    Science.gov (United States)

    Lee, Eujin; Kim, Sun-Gun; Park, Na-Young; Park, Hyo-Hyun; Jeong, Kyu-Tae; Choi, Jongkeun; Lee, In-Hae; Lee, Hwadong; Lee, Eunkyung

    2017-01-01

    Background: A Korean herbal medicine, KOTMIN13, composed of Inula japonica Thunberg, Trichosanthes kirilowii Maximowicz var. japonica kitamura, Peucedanum praeruptorum Dunn, and Allium macrostemon Bge, has been used for anti-allergic and anti-asthmatic treatment in oriental clinics, but its activity has not been investigated. Materials and Methods: To evaluate the anti-inflammatory activity of KOTMIN13 for in vitro study, LPS-stimulated RAW 264.7 cells were used to induce the production and expression of inflammatory mediators and its mechanisms. 12-O-Tetradecanoylphorobol-13 aceate (TPA)-induced ear edema and carrageenan-induced paw edema models were also used to evaluate the effect of KOTMIN13 on acute inflammation in vivo. Results: KOTMIN13 reduced the release of inflammatory mediators [nitric oxide, prostaglandin E2, interleukin (IL)-1β, and IL-6] and the protein expression of inducible nitric oxide synthase and cyclooxygenase-2 in LPS-stimulated RAW 264.7 cells. Mechanism studies showed the attenuation of LPS-induced NF-κB activation by KOTMIN13 via IκBα degradation abrogation and a subsequent decrease in nuclear p65 levels. Activation of mitogen-activated protein kinases (ERK, JNK, and p38) was also suppressed. Furthermore, KOTMIN13 ameliorated the development of TPA-induced ear edema and carrageenan-induced paw edema in acute inflammatory edema mouse models. Conclusion: Our study demonstrates that KOTMIN13 inhibits inflammatory mediators through the inhibitions of NF-κB and MAPK activities in LPS-induced RAW 264.7 cells, as well as acute inflammation in edema models, indicating that KOTMIN13 is an effective suppressor for anti-inflammatory activities. SUMMARY KOTMIN13 decrease the production of No, PGE2, and proinflammatory cytokine (TNF-∝, IL-1β,IL-6).KOTMIN13 Suppressed the degradation of NF-kβ and IKβα and the phosorylation of MAP Kinases.Topical application of KOTMIN13 reduced mouse ear edema.Oral administration of KOTMIN13 decreased

  8. Protective effects of Zn2+against cobalt nanoparticles and cobalt chloride-induced cytotoxicity of RAW 264.7cells via ROS pathway.

    Science.gov (United States)

    Zhu, Hai; Liu, Yake; Hong, Hongxiang; Wang, Wei; Liu, Fan

    2017-04-29

    Recent concerns have emerged surrounding the toxicity that cobalt may represent when used in MOM implants. Owing to corrosion and wear of MOM implants, the subsequent released cobalt nanoparticles (CoNPs) or Co ions (Co 2+ ) can cause adverse reactions, such as the generation of pseudotumors, extensive necrosis, early osteolysis, and implants failure. The present study confirmed that CoNPs and Co 2+ can induce dose- and time-dependent cytotoxicity with increasing reactive oxygen species (ROS) levels. Additionally, using metallothionein (MT), a heavy metal-binding protein, the present study assessed the protective effects of Zn 2+ against CoNPs and Co 2+ -induced cytotoxicity of RAW 264.7 cells through ROS pathway. Further studies are needed to explore the underlying protective mechanisms in vitro. However, the current findings indicate that the ROS pathway may be a potential target for therapeutic interventions. Copyright © 2017. Published by Elsevier Inc.

  9. Protective effect of Hypericum calabricum Sprengel on oxidative damage and its inhibition of nitric oxide in lipopolysaccharide-stimulated RAW 264.7 macrophages

    Directory of Open Access Journals (Sweden)

    Giancarlo A Statti

    2011-01-01

    Full Text Available The present study shows for the first time the phenolic composition and the in vitro properties (antioxidant and inhibition of nitric oxide production of Hypericum calabricum Sprengel collected in Italy. The content of hypericins (hypericin and pseudohypericin, hyperforin, flavonoids (rutin, hyperoside, isoquercetrin, quercitrin, quercetin and biapigenin and chlorogenic acid of H. calabricum, have been determined. The ethyl acetate fraction from the aerial parts of H. calabricum exhibited activity against the radical 1,1-diphenyl-2-picrylhydrazyl (DPPH with IC50 value of 1.6 jig/ml. The test for inhibition of nitric oxide (NO production was performed using the murine monocytic macrophage cell line RAW 264.7. The ethyl acetate fraction had significant activity with an IC50 value of 102 jig/ml and this might indicate that it would have an anti-inflammatory effect in vivo.

  10. Synthesis and evaluation of new α-methylene-γ-lactone carbamates with NO production inhibitory effects in lipopolysaccharide-induced RAW 264.7 macrophages.

    Science.gov (United States)

    Shao, Wen-Hao; Chen, Bing-Yang; Cheng, Xiang-Rong; Yuan, Hu; Chen, Hao; Chang, Wan-Lin; Ye, Ji; Lin, Shan; Sun, Qing-Yan; Zhang, Wei-Dong

    2015-03-26

    A series of new α-methylene-γ-lactone carbamates were synthesized by an asymmetric synthetic route. The activities on inhibiting nitric oxide (NO) release of these compounds were evaluated in lipopolysaccharide (LPS)-induced RAW 264.7 macrophages. The results indicated that most of the compounds except one exhibited potent NO inhibitory effect with IC50 value more than 2 μΜ. The cytotoxicities of these compounds were estimated via MTT assays. The results suggested that six compounds were accompanied by low cytotoxicity. The structure-activity relationships were also discussed. The S configuration of C3 on lactones ring would be more helpful to NO inhibitory effect. Copyright © 2015 Elsevier Masson SAS. All rights reserved.

  11. Effect of binary combination of some plant-derived molluscicides with MGK-264 or piperonyl butoxide on the reproduction of the snail Lymnaea acuminata.

    Science.gov (United States)

    Singh, Priyamvada; Singh, Vinay K; Singh, Dinesh K

    2005-02-01

    The effects of sub-lethal treatments (20 and 60% of 24-h LC(50)) with plant-derived molluscicides Annona squamosa, acetogenins, Argemone mexicana seed and protopine, in combination (1 + 5) with MGK-264 (ENT 8184) or piperonyl butoxide on the reproduction of Lymnaea acuminata has been studied. The plant-derived molluscicides and their active molluscicidal components, protopine and acetogenins, in combination with ENT 8184 or piperonyl butoxide caused a significant reduction in the fecundity, hatchability and survival of young snails. Combination of A squamosa seed powder with piperonyl butoxide was very effective as it caused a complete arrest of snail fecundity within 24 h of treatment. Removal of the snails to fresh water after the 96-h treatments caused a significant recovery in the fecundity of L acuminata. Copyright (c) 2005 Society of Chemical Industry.

  12. A Sesquiterpene Quinone, 5-Epi-smenospongine, Promotes TNF-α Production in LPS-stimulated RAW 264.7 Cells

    Directory of Open Access Journals (Sweden)

    Masataka Mochizuki

    2007-10-01

    Full Text Available Eight sesquiterpene quinones: ilimaquinone (1, smenospongidine (3,smenospongiarine (5, smenospongine (7, and their corresponding 5-epimers 2, 4, 6, and 8,isolated from the Palauan marine sponge Hippospongia sp., were examined regarding theireffects on TNF-α production in LPS-stimulated RAW 264.7 cells. 5-Epi-smenospongine(8 promoted the production of TNF-α to a level three times greater than the control at10 μM, but compounds 1-7 did not show apparent activity. The results suggest that thecis-decaline ring and a primary amine in the benzoquinone ring are necessary for activity.This is the first study to report the modulation of TNF-α production by a sesquiterpenequinone.

  13. In vitro evaluation of cytotoxic and inflammatory properties of silica nanoparticles of different sizes in murine RAW 264.7 macrophages

    International Nuclear Information System (INIS)

    Park, Margriet V. D. Z.; Lynch, Iseult; Ramírez-García, Sonia; Dawson, Kenneth A.; Fonteyne, Liset de la; Gremmer, Eric; Slob, Wout; Briedé, Jacob J.; Elsaesser, Andreas; Howard, C. Vyvyan; Loveren, Henk van; Jong, Wim H. de

    2011-01-01

    The biological response to four well-characterized amorphous silica nanoparticles was investigated in RAW 264.7 macrophages in view of their potential application as drug carriers to sites of inflammation. All silica nanoparticles-induced cell membrane damage, reduced metabolic activity, generated ROS and released various cytokines, but to different extents. Two silica nanoparticles of 34 nm (A and B) with different zetapotentials were more cytotoxic than (aggregated) 11 and 248 nm nanoparticles, while cytokines were mostly induced by the (aggregated) 11 nm and only one of the 34 nm nanoparticles (34A). The results indicate that specific silica nanoparticles may have counterproductive effects, for example when used as carriers of anti-inflammatory drugs. The physicochemical properties determining the response of nanoparticles vary for different responses, implying that a screening approach for the safe development of nanoparticles needs to consider the role of combinations of (dynamic) physicochemical properties and needs to include multiple toxicity endpoints.

  14. Relationship between size and surface modification of silica particles and enhancement and suppression of inflammatory cytokine production by lipopolysaccharide- or peptidoglycan-stimulated RAW264.7 macrophages

    International Nuclear Information System (INIS)

    Uemura, Eiichiro; Yoshioka, Yasuo; Hirai, Toshiro; Handa, Takayuki; Nagano, Kazuya; Higashisaka, Kazuma; Tsutsumi, Yasuo

    2016-01-01

    Although nanomaterials are used in an increasing number of commodities, the relationships between their immunotoxicity and physicochemical properties such as size or surface characteristics are not fully understood. Here we demonstrated that pretreatment with amorphous silica particles (SPs) of various sizes (diameters of 10–1000 nm), with or without amine surface modification, significantly decreased interleukin 6 production by RAW264.7 macrophages following lipopolysaccharide or peptidoglycan stimulation. Furthermore, nanosized, but not microsized, SPs significantly enhanced tumor necrosis factor-α production in macrophages stimulated with lipopolysaccharide. This altered cytokine response was distinct from the inflammatory responses induced by treatment with the SPs alone. Additionally, the uptake of SPs into macrophages by phagocytosis was found to be crucial for the suppression of macrophage immune response to occur, irrespective of particle size or surface modification. Together, these results suggest that SPs may not only increase susceptibility to microbial infection, but that they may also be potentially effective immunosuppressants.

  15. Relationship between size and surface modification of silica particles and enhancement and suppression of inflammatory cytokine production by lipopolysaccharide- or peptidoglycan-stimulated RAW264.7 macrophages

    Energy Technology Data Exchange (ETDEWEB)

    Uemura, Eiichiro, E-mail: uemura-e@phs.osaka-u.ac.jp; Yoshioka, Yasuo, E-mail: y-yoshioka@biken.osaka-u.ac.jp; Hirai, Toshiro, E-mail: toshiro.hirai@pitt.edu; Handa, Takayuki, E-mail: handa-t@phs.osaka-u.ac.jp; Nagano, Kazuya, E-mail: knagano@phs.osaka-u.ac.jp; Higashisaka, Kazuma, E-mail: higashisaka@phs.osaka-u.ac.jp; Tsutsumi, Yasuo, E-mail: ytsutsumi@phs.osaka-u.ac.jp [Osaka University, Laboratory of Toxicology and Safety Science, Graduate School of Pharmaceutical Sciences (Japan)

    2016-06-15

    Although nanomaterials are used in an increasing number of commodities, the relationships between their immunotoxicity and physicochemical properties such as size or surface characteristics are not fully understood. Here we demonstrated that pretreatment with amorphous silica particles (SPs) of various sizes (diameters of 10–1000 nm), with or without amine surface modification, significantly decreased interleukin 6 production by RAW264.7 macrophages following lipopolysaccharide or peptidoglycan stimulation. Furthermore, nanosized, but not microsized, SPs significantly enhanced tumor necrosis factor-α production in macrophages stimulated with lipopolysaccharide. This altered cytokine response was distinct from the inflammatory responses induced by treatment with the SPs alone. Additionally, the uptake of SPs into macrophages by phagocytosis was found to be crucial for the suppression of macrophage immune response to occur, irrespective of particle size or surface modification. Together, these results suggest that SPs may not only increase susceptibility to microbial infection, but that they may also be potentially effective immunosuppressants.

  16. Non-tables look-up search algorithm for efficient H.264/AVC context-based adaptive variable length coding decoding

    Science.gov (United States)

    Han, Yishi; Luo, Zhixiao; Wang, Jianhua; Min, Zhixuan; Qin, Xinyu; Sun, Yunlong

    2014-09-01

    In general, context-based adaptive variable length coding (CAVLC) decoding in H.264/AVC standard requires frequent access to the unstructured variable length coding tables (VLCTs) and significant memory accesses are consumed. Heavy memory accesses will cause high power consumption and time delays, which are serious problems for applications in portable multimedia devices. We propose a method for high-efficiency CAVLC decoding by using a program instead of all the VLCTs. The decoded codeword from VLCTs can be obtained without any table look-up and memory access. The experimental results show that the proposed algorithm achieves 100% memory access saving and 40% decoding time saving without degrading video quality. Additionally, the proposed algorithm shows a better performance compared with conventional CAVLC decoding, such as table look-up by sequential search, table look-up by binary search, Moon's method, and Kim's method.

  17. High-resolution, low-delay, and error-resilient medical ultrasound video communication using H.264/AVC over mobile WiMAX networks.

    Science.gov (United States)

    Panayides, Andreas; Antoniou, Zinonas C; Mylonas, Yiannos; Pattichis, Marios S; Pitsillides, Andreas; Pattichis, Constantinos S

    2013-05-01

    In this study, we describe an effective video communication framework for the wireless transmission of H.264/AVC medical ultrasound video over mobile WiMAX networks. Medical ultrasound video is encoded using diagnostically-driven, error resilient encoding, where quantization levels are varied as a function of the diagnostic significance of each image region. We demonstrate how our proposed system allows for the transmission of high-resolution clinical video that is encoded at the clinical acquisition resolution and can then be decoded with low-delay. To validate performance, we perform OPNET simulations of mobile WiMAX Medium Access Control (MAC) and Physical (PHY) layers characteristics that include service prioritization classes, different modulation and coding schemes, fading channels conditions, and mobility. We encode the medical ultrasound videos at the 4CIF (704 × 576) resolution that can accommodate clinical acquisition that is typically performed at lower resolutions. Video quality assessment is based on both clinical (subjective) and objective evaluations.

  18. Immunomodulatory activity of enzymatically synthesized glycogen and its digested metabolite in a co-culture system consisting of differentiated Caco-2 cells and RAW264.7 macrophages.

    Science.gov (United States)

    Yasuda, Michiko; Furuyashiki, Takashi; Nakamura, Toshiyuki; Kakutani, Ryo; Takata, Hiroki; Ashida, Hitoshi

    2013-09-01

    Previously, we developed enzymatically synthesized glycogen (ESG) from starch, and showed its immunomodulatory and dietary fiber-like activities. In this study, we investigated the metabolism of ESG and its immunomodulatory activity using differentiated Caco-2 cells as a model of the intestinal barrier. In a co-culture system consisting of differentiated Caco-2 cells and RAW264.7 macrophages, mRNA expression of IL-6, IL-8, IL-1β and BAFF cytokines was up-regulated in Caco-2 cells and IL-8 production in basolateral medium was induced after 24 h apical treatment with 5 mg ml(-1) of ESG. The mRNA level of iNOS was also up-regulated in RAW264.7 macrophages. After characterization of the binding of anti-glycogen monoclonal antibodies (IV58B6 and ESG1A9) to ESG and its digested metabolite resistant glycogen (RG), an enzyme-linked immunosorbent assay (ELISA) system was developed to quantify ESG and RG. Using this system, we investigated the metabolism of ESG in differentiated Caco-2 cells. When ESG (7000 kDa, 5 mg ml(-1)) was added to the apical side of Caco-2 monolayers, ESG disappeared and RG (about 3000 kDa, 3.5 mg ml(-1)) appeared in the apical solution during a 24 h incubation. Neither ESG nor RG was detected in the basolateral solution. In addition, both ESG and RG were bound to TLR2 in Caco-2 cells. In conclusion, we suggest that ESG is metabolized to a RG-like structure in the intestine, and this metabolite activates the immune system via stimulation of the intestinal epithelium, although neither ESG nor its metabolite could permeate the intestinal cells under our experimental conditions. These results provide evidence for the beneficial function of ESG as a food ingredient.

  19. Comparative physicochemical and biological characterization of NIST Interim Reference Material PM2.5 and SRM 1648 in human A549 and mouse RAW264.7 cells.

    Science.gov (United States)

    Mitkus, Robert J; Powell, Jan L; Zeisler, Rolf; Squibb, Katherine S

    2013-12-01

    The epidemiological association between exposure to fine particulate matter (PM2.5) and adverse health effects is well-known. Here we report the size distribution, metals content, endotoxin content, and biological activity of National Institute of Standards and Technology (NIST) Interim Reference Material (RM) PM2.5. Biological activity was measured in vitro by effects on cell viability and the release of four inflammatory immune mediators, from human A549 alveolar epithelial cells or murine RAW264.7 monocytes. A dose range covering three orders of magnitude (1-1000μg/mL) was tested, and biological activity was compared to an existing Standard Reference Material (SRM) for urban PM (NIST SRM 1648). Robust release of IL-8 and MCP-1 from A549 cells was observed in response to IRM PM2.5 exposures. Significant TNF-α, but not IL-6, secretion from RAW264.7 cells was observed in response to both IRM PM2.5 and SRM 1648 particle types. Cytokine or chemokine release at high doses often occurred in the presence of cytotoxicity, likely as a result of externalization of preformed mediator. Our results are consistent with a local cytotoxic and pro-inflammatory mechanism of response to exposure to inhaled ambient PM2.5 and reinforce the continued relevance of in vitro assays for mechanistic research in PM toxicology. Our study furthers the goal of developing reference samples of environmentally relevant particulate matter of various sizes that can be used for hypothesis testing by multiple investigators. Published by Elsevier Ltd.

  20. Ascorbic acid pre-treated quartz stimulates TNF-α release in RAW 264.7 murine macrophages through ROS production and membrane lipid peroxidation

    Directory of Open Access Journals (Sweden)

    Benvenuto Federica

    2009-03-01

    Full Text Available Abstract Background Inhalation of crystalline silica induces a pulmonary fibrotic degeneration called silicosis caused by the inability of alveolar macrophages to dissolve the crystalline structure of phagocytosed quartz particles. Ascorbic acid is capable of partially dissolving quartz crystals, leading to an increase of soluble silica concentration and to the generation of new radical sites on the quartz surface. The reaction is specific for the crystalline forms of silica. It has been already demonstrated an increased cytotoxicity and stronger induction of pro-inflammatory cyclooxygenase-2 (COX-2 by ascorbic acid pre-treated quartz (QA compared to untreated quartz (Q in the murine macrophage cell line RAW 264.7. Methods Taking advantage of the enhanced macrophage response to QA as compared to Q particles, we investigated the first steps of cell activation and the contribution of early signals generated directly from the plasma membrane to the production of TNF-α, a cytokine that activates both inflammatory and fibrogenic pathways. Results Here we demonstrate that TNF-α mRNA synthesis and protein secretion are significantly increased in RAW 264.7 macrophages challenged with QA as compared to Q particles, and that the enhanced response is due to an increase of intracellular ROS. Plasma membrane-particle contact, in the absence of phagocytosis, is sufficient to trigger TNF-α production through a mechanism involving membrane lipid peroxidation and this appears to be even more detrimental to macrophage survival than particle phagocytosis itself. Conclusion Taken together these data suggest that an impairment of pulmonary macrophage phagocytosis, i.e. in the case of alcoholic subjects, could potentiate lung disease in silica-exposed individuals.

  1. Ascorbic acid pre-treated quartz stimulates TNF-alpha release in RAW 264.7 murine macrophages through ROS production and membrane lipid peroxidation.

    Science.gov (United States)

    Scarfì, Sonia; Magnone, Mirko; Ferraris, Chiara; Pozzolini, Marina; Benvenuto, Federica; Benatti, Umberto; Giovine, Marco

    2009-03-19

    Inhalation of crystalline silica induces a pulmonary fibrotic degeneration called silicosis caused by the inability of alveolar macrophages to dissolve the crystalline structure of phagocytosed quartz particles. Ascorbic acid is capable of partially dissolving quartz crystals, leading to an increase of soluble silica concentration and to the generation of new radical sites on the quartz surface. The reaction is specific for the crystalline forms of silica. It has been already demonstrated an increased cytotoxicity and stronger induction of pro-inflammatory cyclooxygenase-2 (COX-2) by ascorbic acid pre-treated quartz (QA) compared to untreated quartz (Q) in the murine macrophage cell line RAW 264.7. Taking advantage of the enhanced macrophage response to QA as compared to Q particles, we investigated the first steps of cell activation and the contribution of early signals generated directly from the plasma membrane to the production of TNF-alpha, a cytokine that activates both inflammatory and fibrogenic pathways. Here we demonstrate that TNF-alpha mRNA synthesis and protein secretion are significantly increased in RAW 264.7 macrophages challenged with QA as compared to Q particles, and that the enhanced response is due to an increase of intracellular ROS. Plasma membrane-particle contact, in the absence of phagocytosis, is sufficient to trigger TNF-alpha production through a mechanism involving membrane lipid peroxidation and this appears to be even more detrimental to macrophage survival than particle phagocytosis itself. Taken together these data suggest that an impairment of pulmonary macrophage phagocytosis, i.e. in the case of alcoholic subjects, could potentiate lung disease in silica-exposed individuals.

  2. Immunomodulatory beta-glucan from Lentinus edodes activates mitogen-activated protein kinases and nuclear factor-kappaB in murine RAW 264.7 macrophages.

    Science.gov (United States)

    Xu, Xiaojuan; Pan, Chen; Zhang, Lina; Ashida, Hitoshi

    2011-09-09

    Lentinan, a cell wall β-glucan from the fruiting bodies of Lentinus edodes, is well known to be a biological defense modifier, but the signal transduction pathway(s) induced by Lentinan have not been elucidated. In this study, we extracted Lentinan (LNT-S) by ultrasonication from Lentinus edodes and report that, in murine RAW 264.7 macrophages, LNT-S glucan activated NF-κB p65 and triggered its nuclear translocation as determined by Western blotting. Moreover, LNT-S enhanced NF-κB-luciferase activity in the Dual-Luciferase gene system assay. Its upstream signaling molecules, MAPKs such as ERK1/2 and JNK1/2, were shown to be activated by assessing the level of phosphorylation in a time- and concentration-dependent manner, but its downstream proinflammatory enzyme, inducible NOS, was not observed. The data evaluated using a TNF-α ELISA kit and Griess reagent further demonstrated that no proinflammatory mediators such as TNF-α and NO were produced by LNT-S stimulation in RAW 264.7 cells. In contrast, LPS significantly induced inducible NOS expression and increased NO and TNF-α production, which are associated with activation of the NF-κB p65/p50 heterodimer complex. It is possible that LNT-S did not activate NF-κB p65/p50, and the activation of NF-κB p65 was not sufficient to stimulate cytokine production. These data demonstrate that LNT-S glucan carries out its immunomodulating activity by activating MAPK signaling pathways without secretion of TNF-α and NO.

  3. Ascorbic acid pre-treated quartz stimulates TNF-α release in RAW 264.7 murine macrophages through ROS production and membrane lipid peroxidation

    Science.gov (United States)

    Scarfì, Sonia; Magnone, Mirko; Ferraris, Chiara; Pozzolini, Marina; Benvenuto, Federica; Benatti, Umberto; Giovine, Marco

    2009-01-01

    Background Inhalation of crystalline silica induces a pulmonary fibrotic degeneration called silicosis caused by the inability of alveolar macrophages to dissolve the crystalline structure of phagocytosed quartz particles. Ascorbic acid is capable of partially dissolving quartz crystals, leading to an increase of soluble silica concentration and to the generation of new radical sites on the quartz surface. The reaction is specific for the crystalline forms of silica. It has been already demonstrated an increased cytotoxicity and stronger induction of pro-inflammatory cyclooxygenase-2 (COX-2) by ascorbic acid pre-treated quartz (QA) compared to untreated quartz (Q) in the murine macrophage cell line RAW 264.7. Methods Taking advantage of the enhanced macrophage response to QA as compared to Q particles, we investigated the first steps of cell activation and the contribution of early signals generated directly from the plasma membrane to the production of TNF-α, a cytokine that activates both inflammatory and fibrogenic pathways. Results Here we demonstrate that TNF-α mRNA synthesis and protein secretion are significantly increased in RAW 264.7 macrophages challenged with QA as compared to Q particles, and that the enhanced response is due to an increase of intracellular ROS. Plasma membrane-particle contact, in the absence of phagocytosis, is sufficient to trigger TNF-α production through a mechanism involving membrane lipid peroxidation and this appears to be even more detrimental to macrophage survival than particle phagocytosis itself. Conclusion Taken together these data suggest that an impairment of pulmonary macrophage phagocytosis, i.e. in the case of alcoholic subjects, could potentiate lung disease in silica-exposed individuals. PMID:19298665

  4. Comparison of the LTQ-Orbitrap Velos and the Q-Exactive for proteomic analysis of 1-1000 ng RAW 264.7 cell lysate digests.

    Science.gov (United States)

    Sun, Liangliang; Zhu, Guijie; Dovichi, Norman J

    2013-01-15

    There is interest in extending bottom-up proteomics to the smallest possible sample size. We investigated the performance of two modern mass spectrometers for the analysis of samples ranging from 1 ng to 1 µg of RAW 264.7 cell lysate digests. An ultra-performance liquid chromatography (UPLC) system coupled with either an LTQ-Orbitrap Velos or a Q-Exactive mass spectrometer was used for peptide separation and identification. For 1-1000 ng RAW 264.7 cell lysate digests, the Q-Exactive generated 10-83% more protein groups and 11-109% more peptides than the LTQ-Orbitrap Velos (higher-energy collisional dissociation, HCD) with MASCOT database searching, due to its faster scan rate and higher resolution. In addition, HCD and collision-induced dissociation (CID) modes of the LTQ-Orbitrap Velos were compared. HCD produced higher peptide and protein group IDs than CID for 1-1000 ng RAW 264.7 cell lysate digests with MASCOT database searching. Database searching results from SEQUEST and MASCOT were also compared and comparable protein group IDs were obtained from the two search engines. The Q-Exactive outperformed the LTQ-Orbitrap Velos for shotgun proteomics analysis of 1 to 1000 ng RAW 264.7 cell lysate digests in terms of obtained peptide and protein group IDs. Copyright © 2012 John Wiley & Sons, Ltd.

  5. Stimulation of Alpha7 Nicotinic Acetylcholine Receptor Attenuates Nicotine-Induced Upregulation of MMP, MCP-1, and RANTES through Modulating ERK1/2/AP-1 Signaling Pathway in RAW264.7 and MOVAS Cells

    Directory of Open Access Journals (Sweden)

    Liping Liu

    2017-01-01

    Full Text Available Vagus nerve stimulation through alpha7 nicotine acetylcholine receptors (α7-nAChR signaling had been demonstrated attenuation of inflammation. This study aimed to determine whether PNU-282987, a selective α7-nAChR agonist, affected activities of matrix metalloproteinase (MMP and inflammatory cytokines in nicotine-treatment RAW264.7 and MOVAS cells and to assess the underlying molecular mechanisms. RAW264.7 and MOVAS cells were treated with nicotine at different concentrations (0, 1, 10, and 100 ng/ml for 0–120 min. Nicotine markedly stimulated the phosphorylation of extracellular signal-regulated kinase1/2 (ERK1/2 and c-Jun in RAW264.7 cells. Pretreatment with U0126 significantly suppressed phosphorylation of ERK1/2 and further attenuated nicotine-induced activation of c-Jun and upregulation of MMP-2, MMP-9, monocyte chemotactic protein- (MCP- 1, and regulated upon activation normal T cell expressed and secreted (RANTES. Similarly, nicotine treatment also increased phosphorylation of c-Jun and expressions of MMP-2, MMP-9, MCP-1, and RANTES in MOVAS cells. When cells were pretreated with PNU-282987, nicotine-induced activations of ERK1/2 and c-Jun in RAW264.7 cells and c-Jun in MOVAS cells were effectively inhibited. Furthermore, nicotine-induced secretions of MMP-2, MMP-9, MCP-1, and RANTES were remarkably downregulated. Treatment with α7-nAChR agonist inhibits nicotine-induced upregulation of MMP and inflammatory cytokines through modulating ERK1/2/AP-1 signaling in RAW264.7 cells and AP-1 in MOVAS cells, providing a new therapeutic for abdominal aortic aneurysm.

  6. IL-17 intensifies IFN-γ-induced NOS2 upregulation in RAW 264.7 cells by further activating STAT1 and NF-κB.

    Science.gov (United States)

    Gao, Qiyue; Liu, Yang; Wu, Yan; Zhao, Qiang; Wang, Lijun; Gao, Shanshan; Wen, Wen; Zhang, Weiping; Guo, Ning; Zhou, Juan; Yuan, Zuyi

    2016-02-01

    Interleukin-17 (IL-17) is a signature cytokine of Th17 cells. Previous research has indicated that IL-17 plays a proinflammatory role by exacerbating interferon-γ (IFN-γ)-induced inflammation. However, prior to this study, it was not known whether inducible nitric oxide synthase (iNOS or NOS2), a signature molecule of inflammation, could be intensified by IL-17 when combined with IFN-γ. Thus, we explored the roles and underlying mechanisms of IL-17 and IFN-γ in the regulation of NOS2 expression in RAW 264.7 cells using qPCR, western blot analysis, colorimetric analysis, ChIP assay and statistical analysis. Although IL-17 alone did not induce NOS2 expression or nitric oxide (NO) production, as shown by western blot analysis and colorimetric analysis, it intensified IFN-γ-induced NOS2 upregulation and NO production in RAW 264.7 cells. The alteration of relevant transcription factors demonstrated that a combination of IFN-γ and IL-17 enhanced Tyr701-phosphorylated signal transducer and activator of transcription 1 [p-STAT1(Y701)] and nuclear factor-κB (NF-κB) activation, nuclear translocations and their binding to the NOS2 promoter, compared with IFN-γ alone, as illustrated by the results of the western blot analysis and ChIP assay. Also, using the corresponding inhibitors of STAT1 and NF-κB, we noted downregulation of the expression of NOS2 induced by IFN-γ alone or in combination with IL-17, respectively. In addition, IFN-γ increased phosphorylated (p-)p38 mitogen-activated protein kinase (MAPK), and accelerated the activation of the NF-κB pathway and the expression of NOS2, but phosphorylated extracellular signal-regulated kinase 1/2 (p-ERK1/2) was reduced by treatment with IFN-γ and IL-17. IL-17 intensified the activation of the NF-κB pathway and NOS2 upregulation induced by IFN-γ by increasing the phosphorylation of p38 MAPK and limiting the phosphorylation of ERK1/2. Taken together, these results suggest that IL-17 intensified IFN-γ-induced NOS2

  7. Anti-inflammatory and heme oxygenase-1 inducing activities of lanostane triterpenes isolated from mushroom Ganoderma lucidum in RAW264.7 cells

    International Nuclear Information System (INIS)

    Choi, Solip; Nguyen, Van Thu; Tae, Nara; Lee, Suhyun; Ryoo, Sungwoo; Min, Byung-Sun; Lee, Jeong-Hyung

    2014-01-01

    Ganoderma lucidum is a popular medicinal mushroom used in traditional medicine for preventing or treating a variety of diseases. In the present study, we investigated the anti-inflammatory and heme oxygenase (HO)-1 inducing effects of 12 lanostane triterpenes from G. lucidum in RAW264.7 cells. Of these, seven triterpenes, butyl lucidenateE 2 , butyl lucidenateD 2 (GT-2), butyl lucidenate P, butyl lucidenateQ, Ganoderiol F, methyl ganodenate J and butyl lucidenate N induced HO-1 expression and suppressed lipopolysaccharide (LPS)-induced nitric oxide (NO) production. Inhibiting HO-1 activity abrogated the inhibitory effects of these triterpenes on the production of NO in LPS-stimulated RAW264.7 cells, suggesting the involvement of HO-1 in the anti-inflammatory effects of these triterpenes. We further studied the anti-inflammatory and HO-1 inducing effects of GT-2. Mitogen-activated protein kinase inhibitors or N-acetylcysteine, an antioxidant, did not suppress GT-2-mediated HO-1 induction; however, LY294002, a phosphoinositide 3-kinase (PI3K) inhibitor, blocked GT-2-induced HO-1 mRNA and protein expression. GT-2 increased nuclear translocation of nuclear factor-E2-related factor 2 (Nrf2) and knockdown of Nrf2 by small interfering RNA blocked GT-2-mediated HO-1 induction, suggesting that GT-2 induced HO-1 expression via the PI3K/AKT-Nrf2 pathway. Consistent with the notion that HO-1 has anti-inflammatory properties, GT-2 inhibited the production of tumor necrosis factor-α and interleukin-6, as well as inducible nitric oxide synthase and cyclooxygenase-2 expression. These findings suggest that HO-1 inducing activities of these lanostane triterpenes may be important in the understanding of a novel mechanism for the anti-inflammatory activity of G. lucidum. - Highlights: • The anti-inflammatory effects of selected triterpenes from Ganoderma lucidum are demonstrated. • Heme oxygenase-1 induction is attributable to the anti-inflammatory properties of these triterpenes

  8. Anti-inflammatory and heme oxygenase-1 inducing activities of lanostane triterpenes isolated from mushroom Ganoderma lucidum in RAW264.7 cells

    Energy Technology Data Exchange (ETDEWEB)

    Choi, Solip [Department of Biochemistry, College of Natural Sciences, Kangwon National University, Chuncheon, Gangwon-Do 200-701 (Korea, Republic of); Nguyen, Van Thu [College of Pharmacy, Catholic University of Daegu, Gyeongsan 712-702 (Korea, Republic of); Tae, Nara; Lee, Suhyun [Department of Biochemistry, College of Natural Sciences, Kangwon National University, Chuncheon, Gangwon-Do 200-701 (Korea, Republic of); Ryoo, Sungwoo [Department of Biological Sciences, College of Natural Sciences, Kangwon National University, Chuncheon, Gangwon-Do 200-701 (Korea, Republic of); Min, Byung-Sun [College of Pharmacy, Catholic University of Daegu, Gyeongsan 712-702 (Korea, Republic of); Lee, Jeong-Hyung, E-mail: jhlee36@kangwon.ac.kr [Department of Biochemistry, College of Natural Sciences, Kangwon National University, Chuncheon, Gangwon-Do 200-701 (Korea, Republic of)

    2014-11-01

    Ganoderma lucidum is a popular medicinal mushroom used in traditional medicine for preventing or treating a variety of diseases. In the present study, we investigated the anti-inflammatory and heme oxygenase (HO)-1 inducing effects of 12 lanostane triterpenes from G. lucidum in RAW264.7 cells. Of these, seven triterpenes, butyl lucidenateE{sub 2}, butyl lucidenateD{sub 2} (GT-2), butyl lucidenate P, butyl lucidenateQ, Ganoderiol F, methyl ganodenate J and butyl lucidenate N induced HO-1 expression and suppressed lipopolysaccharide (LPS)-induced nitric oxide (NO) production. Inhibiting HO-1 activity abrogated the inhibitory effects of these triterpenes on the production of NO in LPS-stimulated RAW264.7 cells, suggesting the involvement of HO-1 in the anti-inflammatory effects of these triterpenes. We further studied the anti-inflammatory and HO-1 inducing effects of GT-2. Mitogen-activated protein kinase inhibitors or N-acetylcysteine, an antioxidant, did not suppress GT-2-mediated HO-1 induction; however, LY294002, a phosphoinositide 3-kinase (PI3K) inhibitor, blocked GT-2-induced HO-1 mRNA and protein expression. GT-2 increased nuclear translocation of nuclear factor-E2-related factor 2 (Nrf2) and knockdown of Nrf2 by small interfering RNA blocked GT-2-mediated HO-1 induction, suggesting that GT-2 induced HO-1 expression via the PI3K/AKT-Nrf2 pathway. Consistent with the notion that HO-1 has anti-inflammatory properties, GT-2 inhibited the production of tumor necrosis factor-α and interleukin-6, as well as inducible nitric oxide synthase and cyclooxygenase-2 expression. These findings suggest that HO-1 inducing activities of these lanostane triterpenes may be important in the understanding of a novel mechanism for the anti-inflammatory activity of G. lucidum. - Highlights: • The anti-inflammatory effects of selected triterpenes from Ganoderma lucidum are demonstrated. • Heme oxygenase-1 induction is attributable to the anti-inflammatory properties of these

  9. Multi-walled carbon nanotubes induce COX-2 and iNOS expression via MAP Kinase-dependent and -independent mechanisms in mouse RAW264.7 macrophages

    Directory of Open Access Journals (Sweden)

    Lee Jong

    2012-05-01

    Full Text Available Abstract Background Carbon nanotubes (CNTs are engineered graphene cylinders with numerous applications in engineering, electronics and medicine. However, CNTs cause inflammation and fibrosis in the rodent lung, suggesting a potential human health risk. We hypothesized that multi-walled CNTs (MWCNTs induce two key inflammatory enzymes in macrophages, cyclooxygenase-2 (COX-2 and inducible nitric oxide synthase (iNOS, through activation of extracellular signal-regulated kinases (ERK1,2. Methods RAW264.7 macrophages were exposed to MWCNTs or carbon black nanoparticles (CBNPs over a range of doses and time course. Uptake and subcellular localization of MWCNTs was visualized by transmission electron microscopy (TEM. Protein levels of COX-2, iNOS, and ERK1,2 (total ERK and phosphorylated ERK were measured by Western blot analysis. Prostaglandin-E2 (PGE2 and nitric oxide (NO levels in cell supernatants were measured by ELISA and Greiss assay, respectively. Results MWCNTs, but not CBNPs, induced COX-2 and iNOS in a time- and dose-dependent manner. COX-2 and iNOS induction by MWCNTs correlated with increased PGE2 and NO production, respectively. MWCNTs caused ERK1,2 activation and inhibition of ERK1,2 (U0126 blocked MWCNT induction of COX-2 and PGE2 production, but did not reduce the induction of iNOS. Inhibition of iNOS (L-NAME did not affect ERK1,2 activation, nor did L-NAME significantly decrease COX-2 induction by MWCNT. Nickel nanoparticles (NiNPs, which are present in MWCNTs as a residual catalyst, also induced COX-2 via ERK-1,2. However, a comparison of COX-2 induction by MWCNTs containing 4.5 and 1.8% Ni did not show a significant difference in ability to induce COX-2, indicating that characteristics of MWCNTs in addition to Ni content contribute to COX-2 induction. Conclusion This study identifies COX-2 and subsequent PGE2 production, along with iNOS induction and NO production, as inflammatory mediators involved in the macrophage response to

  10. Intra Frame Coding In Advanced Video Coding Standard (H.264) to Obtain Consistent PSNR and Reduce Bit Rate for Diagonal Down Left Mode Using Gaussian Pulse

    Science.gov (United States)

    Manjanaik, N.; Parameshachari, B. D.; Hanumanthappa, S. N.; Banu, Reshma

    2017-08-01

    Intra prediction process of H.264 video coding standard used to code first frame i.e. Intra frame of video to obtain good coding efficiency compare to previous video coding standard series. More benefit of intra frame coding is to reduce spatial pixel redundancy with in current frame, reduces computational complexity and provides better rate distortion performance. To code Intra frame it use existing process Rate Distortion Optimization (RDO) method. This method increases computational complexity, increases in bit rate and reduces picture quality so it is difficult to implement in real time applications, so the many researcher has been developed fast mode decision algorithm for coding of intra frame. The previous work carried on Intra frame coding in H.264 standard using fast decision mode intra prediction algorithm based on different techniques was achieved increased in bit rate, degradation of picture quality(PSNR) for different quantization parameters. Many previous approaches of fast mode decision algorithms on intra frame coding achieved only reduction of computational complexity or it save encoding time and limitation was increase in bit rate with loss of quality of picture. In order to avoid increase in bit rate and loss of picture quality a better approach was developed. In this paper developed a better approach i.e. Gaussian pulse for Intra frame coding using diagonal down left intra prediction mode to achieve higher coding efficiency in terms of PSNR and bitrate. In proposed method Gaussian pulse is multiplied with each 4x4 frequency domain coefficients of 4x4 sub macro block of macro block of current frame before quantization process. Multiplication of Gaussian pulse for each 4x4 integer transformed coefficients at macro block levels scales the information of the coefficients in a reversible manner. The resulting signal would turn abstract. Frequency samples are abstract in a known and controllable manner without intermixing of coefficients, it avoids

  11. Silencing of H19 inhibits the adipogenesis and inflammation response in ox-LDL-treated Raw264.7 cells by up-regulating miR-130b.

    Science.gov (United States)

    Han, Yong; Ma, Jie; Wang, Junping; Wang, Liping

    2018-01-01

    Atherosclerosis (AS) is the main cause of cardiovascular diseases (CADs). Lipid accumulation and inflammatory response in macrophages are two key factors in the pathogenesis of AS. In this study, we aimed to explore the regulating role of long non-coding RNA (LncRNA)-H19 in oxygenized low density lipoprotein (ox-LDL)-treated Raw264.7 cells. Compared with the healthy control, a relatively higher level of H19 was detected in the blood samples from AS patients. Obviously up-regulated expression of TG (triglycerides)/TC (total cholesterol)/LDL-C (low density lipoprotein-cholesterol) and down-regulated level of HDL-C (high density lipoprotein-cholesterol) were detected in ox-LDL-treated Raw264.7 cells. Besides that, increased expression of H19 was detected in ox-LDL-treated Raw264.7 cells. To examine the function of H19, gene knockdown was performed using short hairpin RNAs (shRNAs). The expression of TG, TC, LDL-C and HDL-C was detected by enzyme linked immunosorbent assay (Elisa) and the expression of lipolytic genes/lipogenic genes (PPARα, CPT-1/REBP-1c, ACS) was examined through western blot. In combination with the result of oil red O staining, we concluded that H19 shRNA effectively decreased lipid accumulation in ox-LDL-treated Raw264.7 cells. Besides that, H19 shRNA decreased the level of pro-inflammatory factors (TNF-α, IL-1β)/CD68+ cells and increased the level of anti-inflammatory factors (IL-4, IL-10)/CD163+ cells compared with the control group. Combined the bioinformatics analyses/luciferase assay with the promoting effect of H19 shRNA on the expression of miR-130b, we speculated that miR-130b was a target of H19 in ox-LDL-treated Raw264.7 cells. Moreover, the adding of LncRNA H19 abolished the facilitating effect of miR-130b inhibitor on adipogenesis and inflammation response by up-regulating the expression of miR-130b. Taken together, our research indicated a H19-miR130b pathway in regulating lipid metabolism and inflammation response in ox

  12. Suppressive effects of extracts from the aerial part of Coriandrum sativum L. on LPS-induced inflammatory responses in murine RAW 264.7 macrophages.

    Science.gov (United States)

    Wu, Trang-Tiau; Tsai, Chia-Wen; Yao, Hsien-Tsung; Lii, Chong-Kuei; Chen, Haw-Wen; Wu, Yu-Ling; Chen, Pei-Yin; Liu, Kai-Li

    2010-08-30

    Coriandrum sativum is used not only as a spice to aid flavour and taste values in food, but also as a folk medicine in many countries. Since little is known about the anti-inflammatory ability of the aerial parts (stem and leaf) of C. sativum, the present study investigated the effect of aerial parts of C. sativum on lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. We further explored the molecular mechanism underlying these pharmacological properties of C. sativum. Ethanolic extracts from both stem and leaf of C. sativum (CSEE) significantly decreased LPS-induced nitric oxide and prostaglandin E(2) production as well as inducible nitric oxide synthase, cyclooxygenase-2, and pro-interleukin-1beta expression. Moreover, LPS-induced IkappaB-alpha phosphorylation and nuclear p65 protein expression as well as nuclear factor-kappaB (NF-kappaB) nuclear protein-DNA binding affinity and reporter gene activity were dramatically inhibited by aerial parts of CSEE. Exogenous addition of CSEE stem and leaf significantly reduced LPS-induced expression of phosphorylated mitogen-activated protein kinases (MAPKs). Our data demonstrated that aerial parts of CSEE have a strong anti-inflammatory property which inhibits pro-inflammatory mediator expression by suppressing NF-kappaB activation and MAPK signal transduction pathway in LPS-induced macrophages. Copyright (c) 2010 Society of Chemical Industry.

  13. Commercial Honeybush (Cyclopia spp. Tea Extract Inhibits Osteoclast Formation and Bone Resorption in RAW264.7 Murine Macrophages—An in vitro Study

    Directory of Open Access Journals (Sweden)

    Amcois Visagie

    2015-10-01

    Full Text Available Honeybush tea, a sweet tasting caffeine-free tea that is indigenous to South Africa, is rich in bioactive compounds that may have beneficial health effects. Bone remodeling is a physiological process that involves the synthesis of bone matrix by osteoblasts and resorption of bone by osteoclasts. When resorption exceeds formation, bone remodeling can be disrupted resulting in bone diseases such as osteoporosis. Osteoclasts are multinucleated cells derived from hematopoietic precursors of monocytic lineage. These precursors fuse and differentiate into mature osteoclasts in the presence of receptor activator of NF-kB ligand (RANKL, produced by osteoblasts. In this study, the in vitro effects of an aqueous extract of fermented honeybush tea were examined on osteoclast formation and bone resorption in RAW264.7 murine macrophages. We found that commercial honeybush tea extract inhibited osteoclast formation and TRAP activity which was accompanied by reduced bone resorption and disruption of characteristic cytoskeletal elements of mature osteoclasts without cytotoxicity. Furthermore, honeybush tea extract decreased expression of key osteoclast specific genes, matrix metalloproteinase-9 (MMP-9, tartrate resistant acid phosphatase (TRAP and cathepsin K. This study demonstrates for the first time that honeybush tea may have potential anti-osteoclastogenic effects and therefore should be further explored for its beneficial effects on bone.

  14. Polar lipids from the marine macroalga Palmaria palmata inhibit lipopolysaccharide-induced nitric oxide production in RAW264.7 macrophage cells.

    Science.gov (United States)

    Banskota, Arjun H; Stefanova, Roumiana; Sperker, Sandra; Lall, Santosh P; Craigie, James S; Hafting, Jeff T; Critchley, Alan T

    2014-05-01

    The EtOAc soluble fraction of a MeOH/CHCl3 extract of Palmaria palmata showed strong nitric oxide (NO) inhibitory activity against lipopolysaccharide (LPS)-induced NO production in murine RAW264.7 cells. NO inhibition-guided isolation led to identification of three new polar lipids including a sulfoquinovosyl diacylglycerol (SQDG) (2S)-1-O-eicosapentaenoyl-2-O-myristoyl-3-O-(6-sulfo-α-D-quinovopyranosyl)-glycerol (1) and two phosphatidylglycerols, 1-O-eicosapentaenoyl-2-O-trans-3-hexadecenoyl-3-phospho-(1'-glycerol)-glycerol (3) and 1-O-eicosapentaenoyl-2-O-palmitoyl-3-phospho-(1'-glycerol)-glycerol (4) from the EtOAc fraction. Seven known lipids were also isolated including a SQDG (2), a phospholipid (5) and five galactolipids (6-10). Structures of the isolated lipids were elucidated by spectral analyses. The isolated SQDGs, phosphatidylglycerols and phospholipid possessed strong and dose-dependent NO inhibitory activity compared to N(G)-methyl-L-arginine acetate salt (L-NMMA), a well-known NO inhibitor used as a positive control. Further study suggested that these polar lipids suppressed NO production through down-regulation of inducible nitric oxide synthase (iNOS). Crown Copyright © 2014. Published by Elsevier Ltd. All rights reserved.

  15. A study of the effects of citrate-coated silver nanoparticles on RAW 264.7 cells using a toolbox of cytotoxic endpoints

    Science.gov (United States)

    Bastos, V.; Duarte, I. F.; Santos, C.; Oliveira, H.

    2017-05-01

    Citrate-coated silver nanoparticles (citrate-AgNPs) are among the most commonly used nanomaterials, widely present in industrial and biomedical products. In this study, the cytotoxicity of 30-nm citrate-AgNPs on the macrophage cell line RAW 264.7 was evaluated, using a battery of cytotoxicity endpoints (viability, oxidative stress, and cytostaticity/clastogenicity), at 24 and 48 h of exposure. Citrate-AgNPs decreased cell proliferation and viability only at 75 μg/mL, suggesting a low sensitivity of RAW cells to lower doses of these AgNPs. After 24 h of exposure, ROS content decreased in cells exposed to 60 μg/mL AgNPs (IC20 value), corroborating the high tolerance of these cells to citrate-AgNPs. However, these cells suffered an impairment of the cell cycle, shown by an increase at the sub-G1 phase. This increase of the sub-G1 population was correlated with an increase of DNA fragmentation, suggesting an increase of apoptosis. Thus, our data are important to understand the effects of low concentrations (IC20) of citrate-AgNPs on in vitro vital macrophage functions.

  16. Single-Cell Mass Spectrometry Reveals Changes in Lipid and Metabolite Expression in RAW 264.7 Cells upon Lipopolysaccharide Stimulation

    Science.gov (United States)

    Yang, Bo; Patterson, Nathan Heath; Tsui, Tina; Caprioli, Richard M.; Norris, Jeremy L.

    2018-03-01

    It has been widely recognized that individual cells that exist within a large population of cells, even if they are genetically identical, can have divergent molecular makeups resulting from a variety of factors, including local environmental factors and stochastic processes within each cell. Presently, numerous approaches have been described that permit the resolution of these single-cell expression differences for RNA and protein; however, relatively few techniques exist for the study of lipids and metabolites in this manner. This study presents a methodology for the analysis of metabolite and lipid expression at the level of a single cell through the use of imaging mass spectrometry on a high-performance Fourier transform ion cyclotron resonance mass spectrometer. This report provides a detailed description of the overall experimental approach, including sample preparation as well as the data acquisition and analysis strategy for single cells. Applying this approach to the study of cultured RAW264.7 cells, we demonstrate that this method can be used to study the variation in molecular expression with cell populations and is sensitive to alterations in that expression that occurs upon lipopolysaccharide stimulation. [Figure not available: see fulltext.

  17. The Effect of the Aerial Part of Lindera akoensis on Lipopolysaccharides (LPS-Induced Nitric Oxide Production in RAW264.7 Cells

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    Yen-Hsueh Tseng

    2013-04-01

    Full Text Available Four new secondary metabolites, 3α-((E-Dodec-1-enyl-4β-hydroxy-5β-methyldihydrofuran-2-one (1, linderinol (6, 4'-O-methylkaempferol 3-O-α-L-(4''-E-p-coumaroylrhamnoside (11 and kaempferol 3-O-α-L-(4''-Z-p-coumaroylrhamnoside (12 with eleven known compounds—3-epilistenolide D1 (2, 3-epilistenolide D2 (3, (3Z,4α,5β-3-(dodec-11-ynylidene-4-hydroxy-5-methylbutanolide (4, (3E,4β,5β-3-(dodec-11-ynylidene-4-hydroxy-5-methylbutanolide (5, matairesinol (7, syringaresinol (8, (+-pinoresinol (9, salicifoliol (10, 4''-p-coumaroylafzelin (13, catechin (14 and epicatechin (15—were first isolated from the aerial part of Lindera akoensis. Their structures were determined by detailed analysis of 1D- and 2D-NMR spectroscopic data. All of the compounds isolated from Lindera akoensis showed that in vitro anti-inflammatory activity decreases the LPS-stimulated production of nitric oxide (NO in RAW 264.7 cell, with IC50 values of 4.1–413.8 µM.

  18. Inhibition of nitric oxide production in LPS-stimulated RAW 264.7 cells by stem bark of Ulmus pumila L.

    Science.gov (United States)

    Joo, Taewoo; Sowndhararajan, Kandhasamy; Hong, Sunghyun; Lee, Jaehak; Park, Sun-Young; Kim, Songmun; Jhoo, Jin-Woo

    2014-01-01

    This study was designed to isolate and identify a potent inhibitory compound against nitric oxide (NO) production from the stem bark of Ulmus pumila L. Ethyl acetate fraction of hot water extract registered a higher level of total phenolics (756.93 mg GAE/g) and also showed strong DPPH (IC50 at 5.6 μg/mL) and ABTS (TEAC value 0.9703) radical scavenging activities than other fractions. Crude extract and its fractions significantly decreased nitrite accumulation in LPS-stimulated RAW 264.7 cells indicating that they potentially inhibited the NO production in a concentration dependent manner. Based on higher inhibitory activity, the ethyl acetate fraction was subjected to Sephadex LH-20 column chromatography and yielded seven fractions and all these fractions registered appreciable levels of inhibitory activity on NO production. The most effective fraction F1 was further purified and subjected to 1H, 13C-NMR and mass spectrometry analysis and the compound was identified as icariside E4. The results suggest that the U. pumila extract and the isolated compound icariside E4 effectively inhibited the NO production and may be useful in preventing inflammatory diseases mediated by excessive production of NO. PMID:25313277

  19. Structural characterization of polysaccharides from Cabernet Franc, Cabernet Sauvignon and Sauvignon Blanc wines: Anti-inflammatory activity in LPS stimulated RAW 264.7 cells.

    Science.gov (United States)

    Bezerra, Iglesias de Lacerda; Caillot, Adriana Rute Cordeiro; Palhares, Lais Cristina Gusmão Ferreira; Santana-Filho, Arquimedes Paixão; Chavante, Suely Ferreira; Sassaki, Guilherme Lanzi

    2018-04-15

    The structural characterization of the polysaccharides and in vitro anti-inflammatory properties of Cabernet Franc (WCF), Cabernet Sauvignon (WCS) and Sauvignon Blanc (WSB) wines were studied for the first time in this work. The polysaccharides of wines gave rise to three fractions of polysaccharides, namely (WCF) 0.16%, (WCS) 0.05% and (WSB) 0.02%; the highest one was chosen for isolation of polysaccharides (WCF). It was identified the presence of mannan, formed by a sequence of α-d-Manp (1 → 6)-linked and side chains O-2 substituted for α-d-mannan (1 → 2)-linked; type II arabinogalactan, formed by (1 → 3)-linked β-d-Galp main chain, substituted at O-6 by (1 → 6)-linked β-d-Galp side chains, and nonreducing end-units of arabinose 3-O-substituted; type I rhamnogalacturonan formed by repeating (1 → 4)-α-d-GalpA-(1 → 2)-α-L-Rhap groups; and traces of type II rhamnogalacturonan. The polysaccharide mixture and isolated fractions inhibited the production of inflammatory cytokines (TNF-α and IL-1β) and mediator (NO) in RAW 264.7 cells stimulated with LPS. Copyright © 2018 Elsevier Ltd. All rights reserved.

  20. Identification of rotundic acid metabolites after oral administration to rats and comparison with the biotransformation by Syncephalastrum racemosum AS 3.264.

    Science.gov (United States)

    Li, Hui; Yang, Bao; Cao, Di; Zhou, Lian; Wang, Qing; Rong, Li; Zhou, Xinghong; Jin, Jing; Zhao, Zhongxiang

    2018-02-20

    The objective of this study was to identify the metabolites of rotundic acid after oral administration to rats and compare the similarities with its biotransformation by Syncephalastrum racemosum AS 3.264 using ultra-high performance liquid chromatography coupled with quadrupole time of flight mass spectrometry. A total of fourteen metabolites were determined based on the mass spectrometry and chromatographic behaviors, among which eleven (M1-M3, M7-M14) and six (M2, M4-M8) metabolites were identified in rats and S. racemosum, respectively. Three identical metabolites (M2, M7 and M8) were found in rats and S. racemosum, indicating that there were metabolic similarities. Moreover, to confirm the results of mass spectrometry, three (M2, M4 and M7) metabolites were obtained by the means of amplifying incubation and their structures were determined by various spectroscopic analyses, and M4 was proved to be a previously undescribed compound. This results showed that in vitro assisted preparation by microbial transformation is a feasible and effective method of obtaining metabolites which are in low amounts and difficult to be prepared in vivo. Copyright © 2017 Elsevier B.V. All rights reserved.

  1. The effect of lipid peroxidation products on reactive oxygen species formation and nitric oxide production in lipopolysaccharide-stimulated RAW 264.7 macrophages.

    Science.gov (United States)

    Ambrozova, Gabriela; Pekarova, Michaela; Lojek, Antonin

    2011-02-01

    Lipid peroxidation induced by oxidants leads to the formation of highly reactive metabolites. These can affect various immune functions, including reactive oxygen species (ROS) and nitric oxide (NO) production. The aim of the present study was to investigate the effects of lipid peroxidation products (LPPs) - acrolein, 4-hydroxynonenal, and malondialdehyde - on ROS and NO production in RAW 264.7 macrophages and to compare these effects with the cytotoxic properties of LPPs. Macrophages were stimulated with lipopolysaccharide (0.1 μg/ml) and treated with selected LPPs (concentration range: 0.1-100 μM). ATP test, luminol-enhanced chemiluminescence, Griess reaction, Western blotting analysis, amperometric and total peroxyl radical-trapping antioxidant parameter assay were used for determining the LPPs cytotoxicity, ROS and NO production, inducible nitric oxide synthase expression, NO scavenging, and antioxidant properties of LPPs, respectively. Our study shows that the cytotoxic action of acrolein and 4-hydroxynonenal works in a dose- and time-dependent manner. Further, our results imply that acrolein, 4-hydroxynonenal, and malondialdehyde can inhibit, to a different degree, ROS and NO production in stimulated macrophages, partially independently of their toxic effect. Also, changes in enzymatic pathways (especially NADPH-oxidase and nitric oxide synthase inhibition) and NO scavenging properties are included in the downregulation of reactive species formation. Copyright © 2010 Elsevier Ltd. All rights reserved.

  2. HO-1 Is Essential for Tetrahydroxystilbene Glucoside Mediated Mitochondrial Biogenesis and Anti-Inflammation Process in LPS-Treated RAW264.7 Macrophages

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    Weihua Yu

    2017-01-01

    Full Text Available 2,3,5,4′-Tetrahydroxystilbene-2-O-β-D-glucoside (TSG, an important monomer extracted from Polygonum multiflorum, can prevent a number of inflammation associated chronic diseases. However, the mechanism involved in TSG inducing anti-inflammatory role remains unclear. As an inducible antioxidant enzyme, Heme oxygenase-1 (HO-1, is crucial for protecting the mammalian cells against adverse stimuli. Here, we found that the TSG treatment strongly induces the expression of HO-1 in an NRF2-depended manner. Meanwhile, TSG increased the mitochondrial mass through upregulation of the mitochondrial biogenesis activators (PGC-1α, NRF1, and TFAM as well as the mitochondrial complex IV. Furthermore, TSG attenuated Lipopolysaccharide (LPS mediated RAW264.7 cells activation and secretion of proinflammatory cytokines, including interleukin-6 (IL-6 and tumor necrosis factor-α (TNF-α. Zinc Protoporphyrin (ZnPP, a selective inhibitor of HO-1 activity, was able to attenuate TSG mediated mitochondrial biogenesis and anti-inflammatory process. Finally, we observed that LPS induced obvious mtDNA depletion and ATP deficiency, which indicated a severe damage of mitochondria. TSG restored the LPS induced mitochondrial dysfunction via activation of the mitochondrial biogenesis. ZnPP treatment markedly reversed the inhibitory effects of TSG on mitochondrial damage and oxidative stress in LPS stimulated macrophages. Taken together, these findings suggest that TSG enhances mitochondrial biogenesis and function mainly via activation the HO-1. TSG can be developed as a potential drug for treatment of inflammatory diseases.

  3. Immunomodulatory Efficacy of Standardized Annona muricata (Graviola) Leaf Extract via Activation of Mitogen-Activated Protein Kinase Pathways in RAW 264.7 Macrophages

    Science.gov (United States)

    2016-01-01

    Annona muricata, commonly known as Graviola, has been utilized as a traditional medicine to treat various human diseases. The aim of this study was to examine the immune-enhancing activity of Graviola leaf extracts in RAW 264.7 macrophage cells. Active ingredients in Graviola leaf extracts (GE) were identified as kaempferol-3-O-rutinoside and quercetin-3-O-rutinoside by LC-MS/MS. When treated with steam or 50% ethanol GE, cell morphology was altered due to initiation of cell differentiation. While the cell viability was not altered by the steam GE, it was reduced by the ethanol GE. Both steam and ethanol GE induced the transcriptional expression of cytokines, including tumor necrosis factor-α (TNF-α) and interleukin-1β, but only the steam extract upregulated inducible nitric oxide synthase (iNOS). In consistence with mRNA expression, the production of TNF-α and nitrite was elevated by both steam and ethanol extracts of Graviola leaves. This is mainly due to activation of mitogen-activated protein (MAP) kinase signaling pathways. These results suggest that Graviola leaves enhance immunity by activation of the MAP kinase pathways. These bioactive properties of Graviola indicate its potential as a health-promoting ingredient to boost the immune system. PMID:28096884

  4. Nogo-B Facilitates LPS-Mediated Immune Responses by Up-Regulation of TLR4-Signaling in Macrophage RAW264.7

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    Ying Zhu

    2017-01-01

    Full Text Available Background/Aims: Nogo-B, a member of the reticulon family of proteins, is mainly located in the endoplasmic reticulum (ER. Here, we investigate the function and mechanism of Nogo-B in the regulation of TLR4-associated immune responses in the macrophage cell line of RAW264.7. Methods: Nogo-B was up- and down-regulated through the use of appropriate adenoviral vectors or siRNA, and the effects of Nogo-B on macrophages under liposaccharide (LPS stimulation were evaluated via western blotting, immunofluorescence, enzyme-linked immunosorbent assay (ELISA, flow cytometric analysis, and transwell assay. Results: Our data indicates that the protein of Nogo-B was down-regulated in a time- and dose-dependent manner following LPS administration in the macrophage. Nogo-B overexpression increased the production of inflammatory cytokines (MCP-1, TNF-α, IL-1β, and TGF-β, enhanced macrophage migration activities, activated major histocompatibility complex II (MHC II, and elevated the expression of macrophage scavenger receptor 1(MSR1, all of which suggest that Nogo-B is necessary for immune responses and plays an important role in regulating macrophage recruitment. Mechanistically, Nogo-B may enhance TLR4 expression in macrophage surfaces, activate mitogen-activated protein kinase (MAPK pathways, and initiate inflammatory responses. Conclusion: These findings illustrate the key regulatory functions of Nogo-B in facilitating LPS-mediated immune responses through promoting the phosphorylation of MAP kinase.

  5. Effect of Three-spot Seahorse Petroleum Ether Extract on Lipopolysaccharide Induced Macrophage RAW264.7 Inflammatory Cytokine Nitric Oxide and Composition Analysis.

    Science.gov (United States)

    Chen, LiPing; Shen, XuanRi; Chen, GuoHua; Cao, XianYing; Yang, Jian

    2015-01-01

    Three-Spot seahorse is a traditional medicine in Asian countries. However, the alcohol extract is largely unknown for its anti-inflammatory activity. This study aimed at elucidating fraction of potent anti-inflammatory activity of seahorse. A systematic solvent extraction method of liquid-liquid fractionation of ethanol crude extract gave four fractions petroleum ether (PE), and ethyl acetate (EtOAc), water saturated butanol (n-BuOH), water (H2O). In this study, PE extract was selected for further study after preliminary screening test, and was connected to silica column chromatography and eluted with different polarity of mobile phases, and obtained four active fractions (Fr I, Fr II, Fr III, Fr IV). Effect of separated fractions on inflammation was investigated in lipopolysaccharide (LPS) stimulated murine RAW264.7 cells in vitro. The result shows that seahorse extract was capable of inhibiting the production of nitric oxide (NO) significantly in a dose dependent manner and exhibited no notable cytotoxicity on cell viability. IC50 of fraction IV was 36.31 μg/mL, indicating that separated fraction possessed potent NO inhibitory activity against LPS-induced inflammatory response, thus, demonstrated its in vitro anti-inflammatory potentiality, it may be at least partially explained by the presence of anti-inflammation active substances, phenolic compounds, phospholipids and polyunsaturated fatty acids, especially phospholipids and polyunsaturated fatty acids. It could be suggested that seahorse lipid-soluble components could be used in functional food and anti-inflammatory drug preparations.

  6. Immunomodulatory Efficacy of Standardized Annona muricata (Graviola Leaf Extract via Activation of Mitogen-Activated Protein Kinase Pathways in RAW 264.7 Macrophages

    Directory of Open Access Journals (Sweden)

    Goon-Tae Kim

    2016-01-01

    Full Text Available Annona muricata, commonly known as Graviola, has been utilized as a traditional medicine to treat various human diseases. The aim of this study was to examine the immune-enhancing activity of Graviola leaf extracts in RAW 264.7 macrophage cells. Active ingredients in Graviola leaf extracts (GE were identified as kaempferol-3-O-rutinoside and quercetin-3-O-rutinoside by LC-MS/MS. When treated with steam or 50% ethanol GE, cell morphology was altered due to initiation of cell differentiation. While the cell viability was not altered by the steam GE, it was reduced by the ethanol GE. Both steam and ethanol GE induced the transcriptional expression of cytokines, including tumor necrosis factor-α (TNF-α and interleukin-1β, but only the steam extract upregulated inducible nitric oxide synthase (iNOS. In consistence with mRNA expression, the production of TNF-α and nitrite was elevated by both steam and ethanol extracts of Graviola leaves. This is mainly due to activation of mitogen-activated protein (MAP kinase signaling pathways. These results suggest that Graviola leaves enhance immunity by activation of the MAP kinase pathways. These bioactive properties of Graviola indicate its potential as a health-promoting ingredient to boost the immune system.

  7. Immunomodulatory Efficacy of Standardized Annona muricata (Graviola) Leaf Extract via Activation of Mitogen-Activated Protein Kinase Pathways in RAW 264.7 Macrophages.

    Science.gov (United States)

    Kim, Goon-Tae; Tran, Nguyen Khoi Song; Choi, Eun-Hye; Song, Yoo-Jeong; Song, Jae-Hwi; Shim, Soon-Mi; Park, Tae-Sik

    2016-01-01

    Annona muricata , commonly known as Graviola, has been utilized as a traditional medicine to treat various human diseases. The aim of this study was to examine the immune-enhancing activity of Graviola leaf extracts in RAW 264.7 macrophage cells. Active ingredients in Graviola leaf extracts (GE) were identified as kaempferol-3-O-rutinoside and quercetin-3-O-rutinoside by LC-MS/MS. When treated with steam or 50% ethanol GE, cell morphology was altered due to initiation of cell differentiation. While the cell viability was not altered by the steam GE, it was reduced by the ethanol GE. Both steam and ethanol GE induced the transcriptional expression of cytokines, including tumor necrosis factor- α (TNF- α ) and interleukin-1 β , but only the steam extract upregulated inducible nitric oxide synthase (iNOS). In consistence with mRNA expression, the production of TNF- α and nitrite was elevated by both steam and ethanol extracts of Graviola leaves. This is mainly due to activation of mitogen-activated protein (MAP) kinase signaling pathways. These results suggest that Graviola leaves enhance immunity by activation of the MAP kinase pathways. These bioactive properties of Graviola indicate its potential as a health-promoting ingredient to boost the immune system.

  8. Hypochoeris radicata attenuates LPS-induced inflammation by suppressing p38, ERK, and JNK phosphorylation in RAW 264.7 macrophages.

    Science.gov (United States)

    Kim, Min-Jin; Kim, Se-Jae; Kim, Sang Suk; Lee, Nam Ho; Hyun, Chang-Gu

    2014-01-01

    Hypochoeris radicata, an invasive plant species, is a large and growing threat to ecosystem integrity on Jeju Island, a UNESCO World Heritage site. Therefore, research into the utilization of H. radicata is important and urgently required in order to solve this invasive plant problem in Jeju Island. The broader aim of our research is to elucidate the biological activities of H. radicata, which would facilitate the conversion of this invasive species into high value-added products. The present study was undertaken to identify the pharmacological effects of H. radicata flower on the production of inflammatory mediators in macrophages. The results indicate that the ethyl acetate fraction of H. radicata extract (HRF-EA) inhibited the production of pro-inflammatory molecules such as NO, iNOS, PGE2, and COX-2, and cytokines such as TNF-α, IL-1ß, and IL-6 in LPS-stimulated RAW 264.7 cells. Furthermore, the phosphorylation of MAPKs such as p38, ERK, and JNK was suppressed by HRF-EA in a concentration-dependent manner. In addition, through HPLC and UPLC fingerprinting, luteolins were also identified and quantified as extract constituents. On the basis of these results, we suggest that H. radicata may be considered possible anti-inflammatory candidates for pharmaceutical and/or cosmetic applications.

  9. Hydroxysafflor Yellow A Inhibits LPS-Induced NLRP3 Inflammasome Activation via Binding to Xanthine Oxidase in Mouse RAW264.7 Macrophages

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    Xiaolong Xu

    2016-01-01

    Full Text Available Hydroxysafflor yellow A (HSYA is an effective therapeutic agent for inflammatory diseases and autoimmune disorders; however, its regulatory effect on NLRP3 inflammasome activation in macrophages has not been investigated. In this study, we predicted the potential interaction between HSYA and xanthine oxidase (XO via PharmMapper inverse docking and confirmed the binding inhibition via inhibitory test (IC50 = 40.04 μM. Computation docking illustrated that, in this HSYA-XO complex, HSYA was surrounded by Leu 648, Leu 712, His 875, Leu 873, Ser 876, Glu 879, Phe 649, and Asn 650 with a binding energy of −5.77 kcal/M and formed hydrogen bonds with the hydroxyl groups of HSYA at Glu 879, Asn 650, and His 875. We then found that HSYA significantly decreased the activity of XO in RAW264.7 macrophages and suppressed LPS-induced ROS generation. Moreover, we proved that HSYA markedly inhibited LPS-induced cleaved caspase-1 activation via suppressing the sensitization of NLRP3 inflammasome and prevented the mature IL-1β formation from pro-IL-1β form. These findings suggest that XO may be a potential target of HSYA via direct binding inhibition and the combination of HSYA-XO suppresses LPS-induced ROS generation, contributing to the depression of NLRP3 inflammasome and inhibition of IL-1β secretion in macrophages.

  10. Differential intensity-dependent effects of pulsed electromagnetic fields on RANKL-induced osteoclast formation, apoptosis, and bone resorbing ability in RAW264.7 cells.

    Science.gov (United States)

    Wang, Pan; Liu, Juan; Yang, Yuefan; Zhai, Mingming; Shao, Xi; Yan, Zedong; Zhang, Xuhui; Wu, Yan; Cao, Lu; Sui, Bingdong; Luo, Erping; Jing, Da

    2017-12-01

    Pulsed electromagnetic fields (PEMF) have been proven to be effective for promoting bone mass and regulating bone turnover both experimentally and clinically. However, the exact mechanisms for the regulation of PEMF on osteoclastogenesis as well as optical exposure parameters of PEMF on inhibiting osteoclastic activities and functions remain unclear, representing significant limitations for extensive scientific application of PEMF in clinics. In this study, RAW264.7 cells incubated with RANKL were exposed to 15 Hz PEMF (2 h/day) at various intensities (0.5, 1, 2, and 3 mT) for 7 days. We demonstrate that bone resorbing capacity was significantly decreased by 0.5 mT PEMF mainly by inhibiting osteoclast formation and maturation, but enhanced at 3 mT by promoting osteoclast apoptosis. Moreover, gene expression of RANK, NFATc1, TRAP, CTSK, BAX, and BAX/BCL-2 was significantly decreased by 0.5 mT PEMF, but increased by 3 mT. Our findings reveal a significant intensity window for low-intensity PEMF in regulating bone resorption with diverse nature for modulating osteoclastogenesis and apoptosis. This study not only enriches our basic knowledge for the regulation of PEMF in osteoclastogenesis, but also may lead to more efficient and scientific clinical application of PEMF in regulating bone turnover and inhibiting osteopenia/osteoporosis. Bioelectromagnetics. 38:602-612, 2017. © 2017 Wiley Periodicals, Inc. © 2017 Wiley Periodicals, Inc.

  11. Anti-Inflammatory Effect of Procyanidins from Wild Grape (Vitis amurensis Seeds in LPS-Induced RAW 264.7 Cells

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    Min-Ji Bak

    2013-01-01

    Full Text Available In the present study, the anti-inflammatory effect and underlying mechanisms of wild grape seeds procyanidins (WGP were examined using lipopolysaccharide- (LPS- stimulated RAW 264.7 cells. We used nitric oxide (NO and prostaglandin E2 (PGE2 and reactive oxygen species (ROS assays to examine inhibitory effect of WGP and further investigated the mechanisms of WGP suppressed LPS-mediated genes and upstream expression by Western blot and confocal microscopy analysis. Our data indicate that WGP significantly reduced NO, PGE2, and ROS production and also inhibited the expression of proinflammatory mediators such as inducible nitric oxide synthase (iNOS and cyclooxygenase-2 (COX-2 protein expressions. Consistently, WGP significantly reduced LPS-stimulated expression of proinflammatory cytokines such as tumor necrosis factor α (TNF-α and interleukin- (IL- 1β. Moreover, WGP prevented nuclear translocation of nuclear factor-κB (NFκB p65 subunit by reducing inhibitory κB-α (IκBα and NFκB phosphorylation. Furthermore, we found that WGP inhibited LPS-induced phosphorylation of p38 mitogen-activated protein kinase (MAPK. Taken together, our results demonstrated that WGP exerts potent anti-inflammatory activity through the inhibition of iNOS and COX-2 by regulating NFκB and p38 MAPK pathway.

  12. [Inhibitory effect of NaV1.9 gene silencing on proliferation, phagocytosis and migration in RAW264.7 cells].

    Science.gov (United States)

    Luo, Yuechen; Zhou, Xin; Ji, Wenjie; Sun, Haiying; Lu, Ruiyi; Jiang, Tiemin; Li, Yuming

    2013-03-01

    To establish the cell line with stable voltage-gated sodium channels (VGSCs/NaVs) α subunit NaV1.9 gene silencing through RNA interference (RNAi) in murine RAW264.7 macrophages, and to investigate proliferation, phagocytosis and migration in this cell line. The stable NaV1.9-deficient cell line was generated by selection in G418 after the transfection of short hairpin (shRNA) plasmid with Lipofectamine TM2000. RNAi efficiency was qualified by RT-PCR; proliferation ability was measured by CCK-8 assay; cell cycle and phagocytic ability were analyzed by flow cytometry; and migrating ability was detected by Transwell migration assay. Stable NaV1.9-deficient cell line was established and the expression of NaV1.9 was reduced by 80%. CCK-8 assay and flow cytometry showed that the proliferation of the NaV1.9-deficient cell line was inhibited (Pmigration assay demonstrated that migrating ability was depressed in the cell line (Pmigration were obviously inhibited.

  13. New sesquiterpene dimers from Inula britannica inhibit NF-kappaB activation and NO and TNF-alpha production in LPS-stimulated RAW264.7 cells.

    Science.gov (United States)

    Jin, Hui Zi; Lee, Dongho; Lee, Jeong Hyung; Lee, Kyeong; Hong, Young-Soo; Choung, Dong-Ho; Kim, Young Ho; Lee, Jung Joon

    2006-01-01

    A bioassay-guided isolation of an ethyl acetate-soluble extract of the aerial parts of Inula britannica var. chinensis (Rupr.) Regel, using an in vitro NF-kappaB reporter gene assay, led to the isolation of four new sesquiterpene dimers bearing a norbornene moiety, inulanolides A-D, and three known sesquiterpenes, 1,6alpha-dihydroxyeriolanolide, 1-acetoxy-6alpha-hydroxyeriolanolide, and eupatolide. The structures of the new compounds were elucidated by spectroscopic methods. Among these compounds, inulanolides B and D and eupatolide, exhibited potent inhibitory activity on the LPS-induced NF-kappaB activation with IC50 values of 0.49 microM, 0.48 microM, and 1.54 microM, respectively. Consistent with their inhibitory effect on NF-kappaB activation, compounds and also strongly inhibited the production of NO and TNF-alpha in the LPS-stimulated RAW264.7 cells with IC50 values in the range of 2 microM.

  14. Inhibitory effect of red ginseng acidic polysaccharide from Korean red ginseng on phagocytic activity and intracellular replication of Brucella abortus in RAW 264.7 cells.

    Science.gov (United States)

    Reyes, Alisha Wehdnesday Bernardo; Simborio, Hannah Leah Tadeja; Hop, Huynh Tan; Arayan, Lauren Togonon; Min, Won Gi; Lee, Hu Jang; Rhee, Man Hee; Chang, Hong Hee; Kim, Suk

    2016-09-30

    Korean red ginseng (KRG) has long been used in traditional Korean and Oriental medicine. However, the anti-bacterial mechanism and therapeutic efficiency of KGR for intracellular Brucella infection are still unclear. In this study, the bactericidal activity of Korean red ginseng acidic polysaccharide (RGAP) on Brucella (B.) abortus and its cytotoxic effects on RAW 264.7 cells were evaluated. In addition, B. abortus internalization and intracellular replication in macrophages were investigated after RGAP treatment. RGAP-incubated cells displayed a marked reduction in the adherence, internalization and intracellular growth of B. abortus in macrophages. Furthermore, decreased F-actin fluorescence was observed relative to untreated B. abortus-infected cells. Western blot analysis of intracellular signaling proteins revealed reduced ERK, JNK and p38α phosphorylation levels in B. abortus-infected RGAP-treated cells compared to the control. Moreover, elevated co-localization of B. abortus-containing phagosomes with lysosome-associated membrane protein 1 (LAMP-1) were observed in RGAP-treated cells compared with the control. Overall, the results of this study suggest that RGAP can disrupt phagocytic activity of B. abortus via suppression of mitogen-activated protein kinases (MAPKs) signaling proteins ERK, JNK and p38 levels and inhibit intracellular replication of B. abortus by enhancing phagolysosome fusion, which may provide an alternative control of brucellosis.

  15. Inhibitory effect of Jeju endemic seaweeds on the production of pro-inflammatory mediators in mouse macrophage cell line RAW 264.7*

    Science.gov (United States)

    Yang, Eun-Jin; Moon, Ji-Young; Kim, Min-Jin; Kim, Dong Sam; Kim, Chan-Shick; Lee, Wook Jae; Lee, Nam Ho; Hyun, Chang-Gu

    2010-01-01

    Seaweed has been used in traditional cosmetics and as a herbal medicine in treatments for cough, boils, goiters, stomach ailments, and urinary diseases, and for reducing the incidence of tumors, ulcers, and headaches. Despite the fact that seaweeds are frequently used in the practice of human health, little is known about the role of seaweed in the context of inflammation. This study aimed to investigate the influence of Jeju endemic seaweed on a mouse macrophage cell line (RAW 264.7) under the stimulation of lipopolysaccharide (LPS). Ethyl acetate extracts obtained from 14 different kinds of Jeju seaweeds were screened for inhibitory effects on pro-inflammatory mediators. Our results revealed that extracts from five seaweeds, Laurencia okamurae, Grateloupia elliptica, Sargassum thunbergii, Gloiopeltis furcata, and Hizikia fusiformis, were potent inhibitors of the production of pro-inflammatory mediators such as nitric oxide (NO), prostaglandin E2 (PGE2), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α). Based on these results, the anti-inflammatory effects and low cell toxicity of these seaweed extracts suggest potential therapeutic applications in the regulation of the inflammatory response. PMID:20443209

  16. Neocryptotanshinone inhibits lipopolysaccharide-induced inflammation in RAW264.7 macrophages by suppression of NF-κB and iNOS signaling pathways

    Directory of Open Access Journals (Sweden)

    Chuanhong Wu

    2015-07-01

    Full Text Available Neocryptotanshinone (NCTS is a natural product isolated from traditional Chinese herb Salvia miltiorrhiza Bunge. In this study, we investigated its anti-inflammatory effects in lipopolysaccharide (LPS-stimulated mouse macrophage (RAW264.7 cells. MTT results showed that NCTS partly reversed LPS-induced cytotoxicity. Real-time PCR results showed that NCTS suppressed LPS-induced mRNA expression of inflammatory cytokines, including tumor necrosis factor α (TNFα, interleukin-6 (IL-6 and interleukin-1β (IL-1β. Moreover, NCTS could decrease LPS-induced nitric oxide (NO production. Western blotting results showed that NCTS could down-regulate LPS-induced expression of inducible nitric oxide synthase (iNOS, p-IκBα, p-IKKβ and p-NF-κB p65 without affecting cyclooxygenase-2 (COX-2. In addition, NCTS inhibited LPS-induced p-NF-κB p65 nuclear translocation. In conclusion, these data demonstrated that NCTS showed anti-inflammatory effect by suppression of NF-κB and iNOS signaling pathways.

  17. Bioactivity-guided isolation of polyacetylenes with inhibitory activity against NO production in LPS-activated RAW264.7 macrophages from the rhizomes of Atractylodes macrocephala.

    Science.gov (United States)

    Yao, Chun-Mei; Yang, Xiu-Wei

    2014-02-03

    The rhizome of Atractylodes macrocephala (Compositae) is one of the most well-known traditional Chinese medicine in China, Japan and Korea, which has a long history of use for the treatment of splenic asthenia, edema, anorexia, and excessive perspiration, etc. As active compounds of anti-inflammatory activity of this medicinal plant have not been fully elucidated, the aim of this study was to isolate and identify the active constituents inhibiting nitric oxide (NO) production from the rhizomes of A. macrocephala. Inhibitory activity against NO production in lipopolysaccharide-activated RAW264.7 macrophages was evaluated by Griess reaction. Fifteen polyacetylenes were isolated from the active ethyl acetate extract using activity-guided screening. The structures of all compounds were elucidated by spectroscopic methods and comparison with published data. The compounds were further tested for their inhibitory activity against NO production. Seven new polyacetylenes, named atractylodemaynes A-G (1-7), along with eight known ones (8-15) were isolated. Compound 14 was isolated for the first time from the rhizomes of A. macrocephala. The study showed that the tested compounds exhibited inhibitory activity against NO production in a dose-dependent manner. Among them, compounds 10, 11 and 12 had relatively stronger inhibitory effect with IC50 values of 28, 23 and 19μM, respectively. The results demonstrated that the polyacetylenes might greatly contribute to the anti-inflammatory activity of the rhizomes of A. macrocephala. © 2013 Elsevier Ireland Ltd. All rights reserved.

  18. Sargachromanol G inhibits osteoclastogenesis by suppressing the activation NF-κB and MAPKs in RANKL-induced RAW 264.7 cells.

    Science.gov (United States)

    Yoon, Weon-Jong; Kim, Kil-Nam; Heo, Soo-Jin; Han, Sang-Chul; Kim, Jihyeon; Ko, Yeong-Jong; Kang, Hee-Kyoung; Yoo, Eun-Sook

    2013-05-17

    Inflammatory cytokines play a major role in osteoclastogenesis, leading to the bone resorption that is frequently associated with osteoporosis. Sargachromanol G (SG), isolated from the brown alga Sargassum siliquastrum, inhibits the production of inflammatory cytokines. In the present study, we determined the effect of SG on receptor activator of NF-κB ligand (RANKL)-induced osteoclast formation. SG inhibited RANKL-induced osteoclast differentiation from RAW264.7 cells without signs of cytotoxicity. Additionally, the expression of osteoclastic marker genes, such as tartrate-resistant acid phosphatase (TRAP), cathepsin K (CTSK), matrix metalloproteinase 9 (MMP9), and calcitonin receptor (CTR), was strongly inhibited. SG inhibited RANKL-induced activation of NF-κB by suppressing RANKL-mediated IκB-α degradation. Furthermore, SG inhibited RANKL-induced phosphorylation of mitogen activated protein kinases (p38, JNK, and ERK). This study identified SG as an inhibitor for osteoclast formation and provided evidence that natural compounds, such as SG, are an alternative medicines for preventing and treating osteolysis. Copyright © 2013 Elsevier Inc. All rights reserved.

  19. Inhibitory effects of constituents from Morus alba var. multicaulis on differentiation of 3T3-L1 cells and nitric oxide production in RAW264.7 cells.

    Science.gov (United States)

    Yang, Zhi-Gang; Matsuzaki, Keiichi; Takamatsu, Satoshi; Kitanaka, Susumu

    2011-07-19

    A new arylbenzofuran, 3',5'-dihydroxy-6-methoxy-7-prenyl-2-arylbenzofuran (1), and 25 known compounds, including moracin R (2), moracin C (3), moracin O (4), moracin P (5), artoindonesianin O (6), moracin D (7), alabafuran A (8), mulberrofuran L (9), mulberrofuran Y (10), kuwanon A (11), kuwanon C (12), kuwanon T (13), morusin (14), kuwanon E (15), sanggenon F (16), betulinic acid (17), uvaol (18), ursolic acid (19), β-sitosterol (20), oxyresveratrol 2-O-β-D-glucopyranoside (21), mulberroside A (22), mulberroside B (23), 5,7-dihydroxycoumarin 7-O-β-D-glucopyranoside (24), 5,7-dihydroxycoumarin 7-O-β-D-apiofuranosyl-(1→6)-O-β-D-glucopyranoside (25) and adenosine (26), were isolated from Morus alba var. multicaulis Perro. (Moraceae). Their structures were determined by spectroscopic methods. The prenyl-flavonoids 11-14, 16, triterpenoids 17,18 and 20 showed significant inhibitory activity towards the differentiation of 3T3-L1 adipocytes. The arylbenzofurans 1-10 and prenyl-flavonoids 11-16 also showed significant nitric oxide (NO) production inhibitory effects in RAW264.7 cells.

  20. Anti-inflammatory effects of egg yolk livetins (α, β, and γ-livetin) fraction and its enzymatic hydrolysates in lipopolysaccharide-induced RAW 264.7 macrophages.

    Science.gov (United States)

    Meram, Chalamaiah; Wu, Jianping

    2017-10-01

    In this study, the egg yolk livetins (α, β, and γ-livetin) fraction and its hydrolysates, prepared by pepsin and Alcalase, were evaluated for their anti-inflammatory effects using lipopolysaccharide (LPS)-induced RAW 264.7 macrophages as an in vitro model. Enzymatic hydrolysis by pepsin and Alcalase successfully transformed the large molecular weight livetins into low molecular mass peptides mostly below 10kDa. Results revealed that livetins and its hydrolysates (peptides) treatment significantly reduced the inflammatory responses as evidenced by inhibition of production of nitric oxide (NO) (22.7-39.2%), pro-inflammatory cytokines such as tumor necrosis factor-α (TNF-α) (36.9-43.2%), interleukin-1β (IL-1β) (26.1-50.9%) and interleukin-6 (IL-6) (60.4-69.0%), and the expression of inducible nitric oxide synthase (iNOS) (58.6-62%). Alcalase hydrolysate showed more effects in inhibiting prostaglandin-E2 (PGE2) production (30.3%) as well as expression of cyclooxygenase-2 (COX-2) (55.7%). In addition, effect of livetins and its hydrolysates on phagocytic capacity of the macrophages was also evaluated. The results indicate that livetins and its enzymatic hydrolysates significantly (pegg yolk livetins and its hydrolysates with anti-inflammatory activity can potentially be used in health food/nutraceutical/pharmaceutical industry for various applications. Copyright © 2017 The Authors. Published by Elsevier Ltd.. All rights reserved.

  1. A study of the effects of citrate-coated silver nanoparticles on RAW 264.7 cells using a toolbox of cytotoxic endpoints

    Energy Technology Data Exchange (ETDEWEB)

    Bastos, V. [University of Aveiro, CESAM, Department of Biology (Portugal); Duarte, I. F. [University of Aveiro, CICECO—Aveiro Institute of Materials, Department of Chemistry (Portugal); Santos, C., E-mail: csantos@fc.up.pt; Oliveira, H., E-mail: holiveira@ua.pt [University of Aveiro, CESAM, Department of Biology (Portugal)

    2017-05-15

    Citrate-coated silver nanoparticles (citrate-AgNPs) are among the most commonly used nanomaterials, widely present in industrial and biomedical products. In this study, the cytotoxicity of 30-nm citrate-AgNPs on the macrophage cell line RAW 264.7 was evaluated, using a battery of cytotoxicity endpoints (viability, oxidative stress, and cytostaticity/clastogenicity), at 24 and 48 h of exposure. Citrate-AgNPs decreased cell proliferation and viability only at 75 μg/mL, suggesting a low sensitivity of RAW cells to lower doses of these AgNPs. After 24 h of exposure, ROS content decreased in cells exposed to 60 μg/mL AgNPs (IC20 value), corroborating the high tolerance of these cells to citrate-AgNPs. However, these cells suffered an impairment of the cell cycle, shown by an increase at the sub-G1 phase. This increase of the sub-G1 population was correlated with an increase of DNA fragmentation, suggesting an increase of apoptosis. Thus, our data are important to understand the effects of low concentrations (IC20) of citrate-AgNPs on in vitro vital macrophage functions.

  2. Effects of Glycated Whey Protein Concentrate on Pro-inflammatory Cytokine Expression and Phagocytic Activity in RAW264.7 Macrophages.

    Science.gov (United States)

    Chun, Su-Hyun; Lee, Hyun Ah; Lee, Keon Bong; Kim, Sae Hun; Park, Kun-Young; Lee, Kwang-Won

    2016-01-01

    The aim of this study was to determine the stimulatory effects of Maillard reaction, a non-enzymatic browning reaction on the expression of pro-inflammatory cytokines and phagocytic activity induced by whey protein concentrate (WPC). Glycated WPC (G-WPC) was prepared by a reaction between WPC and the lactose it contained. The fluorescence intensity of G-WPC dramatically increased after one day, and high molecular weight complexes formed via the Maillard reaction were also observed in the sodium dodecyl sulfate-polyacrylamide gel electrophoresis profiles. G-WPC demonstrated immunomodulatory effects, including stimulation of increased nitric oxide production and cytokine expressions (i.e., tumor necrosis factor-α, interleukin (IL)-1β, and IL-6), compared to WPC. Furthermore, the phagocytic activity of RAW264.7 cells was significantly increased upon treatment with G-WPC, compared to WPC. Therefore, we suggest that G-WPC can be utilized as an improved dietary source for providing immune modulating activity.

  3. Rate-distortion analysis of dead-zone plus uniform threshold scalar quantization and its application--part II: two-pass VBR coding for H.264/AVC.

    Science.gov (United States)

    Sun, Jun; Duan, Yizhou; Li, Jiangtao; Liu, Jiaying; Guo, Zongming

    2013-01-01

    In the first part of this paper, we derive a source model describing the relationship between the rate, distortion, and quantization steps of the dead-zone plus uniform threshold scalar quantizers with nearly uniform reconstruction quantizers for generalized Gaussian distribution. This source model consists of rate-quantization, distortion-quantization (D-Q), and distortion-rate (D-R) models. In this part, we first rigorously confirm the accuracy of the proposed source model by comparing the calculated results with the coding data of JM 16.0. Efficient parameter estimation strategies are then developed to better employ this source model in our two-pass rate control method for H.264 variable bit rate coding. Based on our D-Q and D-R models, the proposed method is of high stability, low complexity and is easy to implement. Extensive experiments demonstrate that the proposed method achieves: 1) average peak signal-to-noise ratio variance of only 0.0658 dB, compared to 1.8758 dB of JM 16.0's method, with an average rate control error of 1.95% and 2) significant improvement in smoothing the video quality compared with the latest two-pass rate control method.

  4. Calmodulin Mediates DNA Repair Pathways Involving H2AX in Response to Low-Dose Radiation Exposure of RAW 264.7 Macrophages

    Energy Technology Data Exchange (ETDEWEB)

    Smallwood, Heather S.; Lopez Ferrer, Daniel; Eberlein, P. Elis; Watson, David J.; Squier, Thomas C.

    2009-02-05

    Understanding the molecular mechanisms that modulate macrophage radioresistance is necessary for the development of effective radiation therapies, as tumor-associated macrophages promote both angiogenesis and matrix remodeling that, in turn, enhance metastasis. In this respect, we have identified a dose-dependent increase in the abundance of the calcium regulatory protein calmodulin (CaM) in RAW 264.7 macrophages upon irradiation. CaM overexpression results in increased macrophage survival following radiation exposure, acting to diminish the sensitivity to low-dose exposures. Increases in CaM abundance also result in an increase in the number of phosphorylated histone H2AX protein complexes associated with DNA repair following macrophage irradiation, with no change in the extent of double-stranded DNA damage. In comparison, when NFκB-dependent pathways are inhibited, through the expression of a dominant-negative IκB construct, there is no significant increase in phosphorylated H2AX upon irradiation. These results indicate that the molecular basis for the up-regulation of histone H2AX mediated DNA-repair pathways is not the result of nonspecific NFκB-dependent pathways or a specific threshold of DNA damage. Rather, increases in CaM abundance act to minimize the low-dose hypersensitivity to radiation to enhance macrophage radioresistance through processes that include the upregulation of DNA repair pathways involving histone protein H2AX phosphorylation.

  5. Frequencies of CCR5-D32, CCR2-64I and SDF1-3’A mutations in Human Immunodeficiency Virus (HIV seropositive subjects and seronegative individuals from the state of Pará in Brazilian Amazonia

    Directory of Open Access Journals (Sweden)

    Fernanda Andreza de Pinho Lott Carvalhaes

    2005-12-01

    Full Text Available The distribution of genetic polymorphisms of chemokine receptors CCR5-delta32, CCR2-64I and chemokine (SDF1-3’A mutations were studied in 110 Human Immunodeficiency Virus type 1 (HIV-1 seropositive individuals (seropositive group and 139 seronegative individuals (seronegative group from the population of the northern Brazilian city of Belém which is the capital of the state of Pará in the Brazilian Amazon. The CCR5-delta32 mutation was found in the two groups at similar frequencies, i.e. 2.2% for the seronegative group and 2.7% for the seropositive group. The frequencies of the SDF1-3’A mutation were 21.0% for the seronegative group and 15.4% for the seropositive group, and the CCR2-64I allele was found at frequencies of 12.5% for the seronegative group and 5.4% for the seropositive group. Genotype distributions were consistent with Hardy-Weinberg expectations in both groups, suggesting that none of the three mutations has a detectable selective effect. Difference in the allelic and genotypic frequencies was statistically significant for the CCR2 locus, the frequency in the seronegative group being twice that found in the seropositive group. This finding may indicate a protective effect of the CCR2-64I mutation in relation to HIV transmission. However, considering that the CCR2-64I mutation has been more strongly associated with a decreased risk for progression for AIDS than to the resistance to the HIV infection, this could reflect an aspect of population structure or a Type I error.

  6. Inhibitory effect of moschamine isolated from Carthamus tinctorius on LPS-induced inflammatory mediators via AP-1 and STAT1/3 inactivation in RAW 264.7 macrophages.

    Science.gov (United States)

    Jo, A-Ra; Han, Hee-Soo; Seo, Seunghwan; Shin, Ji-Sun; Lee, Jae Yeol; Kim, Hyoung Ja; Lee, Kyung-Tae

    2017-12-01

    Seeds of Carthamus tinctorius L. (Compositae) have been used in Korean traditional medicines for the treatment of cardiovascular and bone diseases. In this study, we investigated the anti-inflammatory effects of known serotonin derivatives (1-9) isolated from the ethyl acetate (EtOAc) soluble fraction from the seeds of C. tinctorius. Compound 2, identified as moschamine, most potently inhibited lipopolysaccharide (LPS)-induced production of prostaglandin E 2 (PGE 2 ) and nitric oxide (NO) in RAW 264.7 macrophages. Moschamine concentration-dependently inhibited LPS-induced PGE 2 and NO production in RAW 264.7 macrophages. Consistent with these findings, moschamine suppressed the protein and mRNA levels of cyclooxygenase-2 (COX-2), microsomal prostaglandin E 2 synthase (mPGES)-1, and inducible NO synthase (iNOS), interleukin (IL)-6, and IL-1β. In addition, pretreatment of moschamine significantly inhibited LPS-stimulated the transcriptional activity of activator protein-1 (AP-1) and the phosphorylation of signal transducer and activator of transcription (STAT)1/3 in RAW 264.7 macrophages. Moreover, moschamine inhibited LPS-induced the phosphorylation of p38 mitogen-activated protein kinase (p38) and extracellular signal-regulated kinase (ERK), but it had no effect on c-Jun N-terminal kinase (JNK). These results suggest that the mechanism of anti-inflammatory activity of moschamine is associated with the downregulation of COX-2, mPGES-1, iNOS, IL-6, and IL-1β expression through the suppression of AP-1 and STAT1/3 activation in LPS-induced RAW 264.7 macrophages. Copyright © 2017 Elsevier Ltd. All rights reserved.

  7. Physical and transcript map of the region between D6S264 and D6S149 on chromosome 6q27, the minimal region of allele loss in sporadic epithelial ovarian cancer

    DEFF Research Database (Denmark)

    Liu, Ying; Emilion, Gracy; Mungall, Andrew J

    2002-01-01

    We have previously shown a high frequency of allele loss at D6S193 (62%) on chromosomal arm 6q27 in ovarian tumours and mapped the minimal region of allele loss between D6S297 and D6S264 (3 cM). We isolated and mapped a single non-chimaeric YAC (17IA12, 260-280 kb) containing D6S193 and D6S297...

  8. Pleurotus giganteus (Berk. Karun & Hyde), the giant oyster mushroom inhibits NO production in LPS/H2O2 stimulated RAW 264.7 cells via STAT 3 and COX-2 pathways.

    Science.gov (United States)

    Baskaran, Asweni; Chua, Kek Heng; Sabaratnam, Vikineswary; Ravishankar Ram, Mani; Kuppusamy, Umah Rani

    2017-01-13

    Pleurotus giganteus (Berk. Karunarathna and K.D. Hyde), has been used as a culinary mushroom and is known to have medicinal properties but its potential as an anti-inflammatory agent to mitigate inflammation triggered diseases is untapped. In this study, the molecular mechanism underlying the protective effect of ethanol extract of P. giganteus (EPG) against lipopolysaccharide (LPS) and combination of LPS and hydrogen peroxide (H 2 O 2 )-induced inflammation on RAW 264.7 macrophages was investigated. The effect of EPG on nitric oxide (NO) production as an indicator of inflammation in RAW 264.7 macrophages was estimated based on Griess reaction that measures nitrite level. The expressions of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), NF-kB activating protein (NKAP), signal transducer and activator of transcription 3 protein (STAT 3) and glutathione peroxidase (GPx) genes were assessed using real time reverse transcription polymerase chain reaction (RT-PCR) approach. EPG (10 μg/ml) showed the highest reduction in the LPS-induced NO production in RAW 264.7 macrophages and significantly suppressed (p < 0.05) the expression iNOS, STAT 3 and COX-2. There was a significant increase (p < 0.05) in combination of LPS and H 2 O 2 - induced iNOS production when compared to the LPS-induced iNOS production in RAW 264.7 macrophages and this concurred with the NO production which was attenuated by EPG at 10 μg/ml. A significant (p < 0.05) down regulation was observed in the combination of LPS and H 2 O 2 -induced iNOS and GPx expression by EPG. Our data suggest that the anti-inflammatory activity of EPG is mediated via the suppression of the STAT 3 and COX-2 pathways and can serve as potential endogenous antioxidant stimulant.

  9. An extract of Phellinus linteus grown on germinated brown rice inhibits inflammation markers in RAW264.7 macrophages by suppressing inflammatory cytokines, chemokines, and mediators and up-regulating antioxidant activity.

    Science.gov (United States)

    Park, Hye-Jin; Han, Eun Su; Park, Dong Ki; Lee, Chan; Lee, Ki Won

    2010-12-01

    The immunomodulatory activity of an organic extract of Phellinus linteus grown on slightly germinated brown rice (PBR) was previously demonstrated. Here, we investigated the possible anti-inflammatory activity of the PBR extract by analyzing its effect on the expression of macrophage-derived cytokines, chemokines, and mediator genes that participate in immune and inflammatory responses and diseases. The extract profoundly inhibited the induction of cytokines and chemokines, including tumor necrosis factor-α, chemokine (C-X-C motif) ligand-10, granulocyte-macrophage colony-stimulating factor, and interleukin-6, in lipopolysaccharide (LPS)-stimulated RAW264.7 mouse macrophage cells. It also greatly inhibited LPS-stimulated production of nitric oxide (NO) and prostaglandin E(2) in RAW264.7 cells by suppressing the expression of inducible NO synthase and cyclooxygenase-2. PBR extract inhibited NO production with a twofold lower half-maximal inhibitory concentration value than P. linteus extract. To elucidate the underlying mechanism of action, we examined the effect of the PBR extract on the LPS-induced phosphorylation of mitogen-activated protein kinases (MAPKs) in RAW264.7 cells. PBR extract greatly inhibited extracellular signal-regulated kinase and c-Jun N-terminal kinase phosphorylation and slightly inhibited p38 MAPK phosphorylation. It also significantly increased intracellular glutathione peroxidase activity and heme oxygenase-1 protein expression. Thus, the PBR extract has anti-inflammatory activity in LPS-stimulated RAW264.7 cells by virtue of its ability to suppress the production of inflammatory cytokines and chemokines via inhibition of MAPK activation and up-regulation of antioxidant activities.

  10. 13-hydroxy linoleic acid increases expression of the cholesterol transporters ABCA1, ABCG1 and SR-BI and stimulates apoA-I-dependent cholesterol efflux in RAW264.7 macrophages

    Directory of Open Access Journals (Sweden)

    Kämmerer Ines

    2011-11-01

    Full Text Available Abstract Background Synthetic activators of peroxisome proliferator-activated receptors (PPARs stimulate cholesterol removal from macrophages through PPAR-dependent up-regulation of liver × receptor α (LXRα and subsequent induction of cholesterol exporters such as ATP-binding cassette transporter A1 (ABCA1 and scavenger receptor class B type 1 (SR-BI. The present study aimed to test the hypothesis that the hydroxylated derivative of linoleic acid (LA, 13-HODE, which is a natural PPAR agonist, has similar effects in RAW264.7 macrophages. Methods RAW264.7 macrophages were treated without (control or with LA or 13-HODE in the presence and absence of PPARα or PPARγ antagonists and determined protein levels of LXRα, ABCA1, ABCG1, SR-BI, PPARα and PPARγ and apolipoprotein A-I mediated lipid efflux. Results Treatment of RAW264.7 cells with 13-HODE increased PPAR-transactivation activity and protein concentrations of LXRα, ABCA1, ABCG1 and SR-BI when compared to control treatment (P Conclusion 13-HODE induces cholesterol efflux from macrophages via the PPAR-LXRα-ABCA1/SR-BI-pathway.

  11. Inhibitor of Tec kinase, LFM-A13, decreases pro-inflammatory mediators production in LPS-stimulated RAW264.7 macrophages via NF-κB pathway.

    Science.gov (United States)

    Wang, Fei; Zhang, Wei; Wang, Chao; Fang, Xu; Cheng, Hao; Liu, Sheng; Chen, Xu-Lin

    2017-05-23

    Tec kinase, a prototypical member of the Tec tyrosine kinases family, was shown to mainly govern lymphocyte proliferation. In the present study, we investigated the role of Tec kinase in acute inflammatory response in lipopolysaccharide (LPS) challenge. First, we demonstrate that Tec kinase activity was observed in RAW264.7 macrophages exposed to LPS. Tec and phosphorylated Tec expression were upregulated in a dose- and time-dependent manner after LPS stimulation. LPS increased monocyte chemotactic protein (MCP)-1 secretion and intercellular adhesion molecule (ICAM)-1 expression, and increasing mRNA expression was consistently observed. LPS also induced IκBα phoshporylaytion and its degradation, increased NF-κB p65 phoshporylaytion and translocation to nuclei in RAW264.7 cells. Pretreatment with LFM-A13 decreased LPS-induced cytokines and chemokines production and mRNA levels, blocked NF-κB transactivation. These effects of LPS were also prevented by Tec-siRNA. Additionally, LFM-A13 or Tec-siRNA obviously inhibited LPS-induced TGFβ-activated kinase 1(TAK1) phosphorylation. Taken together, our results suggest that Tec kinase involves in acute inflammation process in LPS-stimulated RAW264.7 cells, at least mediated by activating TAK1/ NF-κB signal pathway.

  12. Cellular mechanisms of the cytotoxic effects of the zearalenone metabolites α-zearalenol and β-zearalenol on RAW264.7 macrophages.

    Science.gov (United States)

    Lu, Jia; Yu, Ji-Yeon; Lim, Shin-Saeng; Son, Young-Ok; Kim, Dong-Hern; Lee, Seung-Ah; Shi, Xianglin; Lee, Jeong-Chae

    2013-04-01

    Zearalenone (ZEN) and its metabolites are commonly found in many food commodities and are known to cause reproductive disorders and genotoxic effects. The major ZEN metabolites are α-zearalenol (α-ZOL) and β-zearalenol (β-ZOL). Although many studies have demonstrated the cytotoxic effects of these metabolites, the mechanisms by which α-ZOL or β-ZOL mediates their cytotoxic effects appear to differ according to cell type and the exposed toxins. We evaluated the toxicity of α-ZOL and β-ZOL on RAW264.7 macrophages and investigated the underlying mechanisms. β-ZOL not only more strongly reduced the viability of cells than did α-ZOL, but it also induced cell death mainly by apoptosis rather than necrosis. The ZEN metabolites induced loss of mitochondrial membrane potential (MMP), mitochondrial changes in Bcl-2 and Bax proteins, and cytoplasmic release of cytochrome c and apoptosis-inducing factor (AIF). Use of an inhibitor specific to c-Jun N-terminal kinase (JNK), p38 kinase or p53, but not pan-caspase or caspase-8, decreased the toxin-induced generation of reactive oxygen species (ROS) and also attenuated the α-ZOL- or β-ZOL-induced decrease of cell viability. Antioxidative enzyme or compounds such as catalase, acteoside, and (E)-1-(3,4-dihydroxyphenethyl)-3-(4-hydroxystyryl)urea suppressed the ZEN metabolite-mediated reduction of cell viability. Further, knockdown of AIF via siRNA transfection diminished the ZEN metabolite-induced cell death. Collectively, these results suggest that the activation of p53, JNK or p38 kinase by ZEN metabolites is the main upstream signal required for the mitochondrial alteration of Bcl-2/Bax signaling pathways and intracellular ROS generation, while MMP loss and nuclear translocation of AIF are the critical downstream events for ZEN metabolite-mediated apoptosis in macrophages. Copyright © 2013 Elsevier Ltd. All rights reserved.

  13. Moringa oleifera Flower Extract Suppresses the Activation of Inflammatory Mediators in Lipopolysaccharide-Stimulated RAW 264.7 Macrophages via NF-κB Pathway

    Science.gov (United States)

    Tan, Woan Sean; Arulselvan, Palanisamy; Karthivashan, Govindarajan; Fakurazi, Sharida

    2015-01-01

    Aim of Study. Moringa oleifera Lam. (M. oleifera) possess highest concentration of antioxidant bioactive compounds and is anticipated to be used as an alternative medicine for inflammation. In the present study, we investigated the anti-inflammatory activity of 80% hydroethanolic extract of M. oleifera flower on proinflammatory mediators and cytokines produced in lipopolysaccharide- (LPS-) induced RAW 264.7 macrophages. Materials and Methods. Cell cytotoxicity was conducted by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Nitric oxide (NO) production was quantified through Griess reaction while proinflammatory cytokines and other key inflammatory markers were assessed through enzyme-linked immunosorbent assay (ELISA) and immunoblotting. Results. Hydroethanolic extract of M. oleifera flower significantly suppressed the secretion and expression of NO, prostaglandin E2 (PGE2), interleukin- (IL-) 6, IL-1β, tumor necrosis factor-alpha (TNF-α), nuclear factor-kappa B (NF-κB), inducible NO synthase (iNOS), and cyclooxygenase-2 (COX-2). However, it significantly increased the production of IL-10 and IκB-α (inhibitor of κB) in a concentration dependent manner (100 μg/mL and 200 μg/mL). Conclusion. These results suggest that 80% hydroethanolic extract of M. oleifera flower has anti-inflammatory action related to its inhibition of NO, PGE2, proinflammatory cytokines, and inflammatory mediator's production in LPS-stimulated macrophages through preventing degradation of IκB-α in NF-κB signaling pathway. PMID:26609199

  14. Mitochondrial function is involved in regulation of cholesterol efflux to apolipoprotein (apoA-I from murine RAW 264.7 macrophages

    Directory of Open Access Journals (Sweden)

    Allen Anne Marie

    2012-12-01

    Full Text Available Abstract Background Mitochondrial DNA damage, increased production of reactive oxygen species and progressive respiratory chain dysfunction, together with increased deposition of cholesterol and cholesteryl esters, are hallmarks of atherosclerosis. This study investigated the role of mitochondrial function in regulation of macrophage cholesterol efflux to apolipoprotein A-I, by the addition of established pharmacological modulators of mitochondrial function. Methods Murine RAW 264.7 macrophages were treated with a range of concentrations of resveratrol, antimycin, dinitrophenol, nigericin and oligomycin, and changes in viability, cytotoxicity, membrane potential and ATP, compared with efflux of [3H]cholesterol to apolipoprotein (apo A-I. The effect of oligomycin treatment on expression of genes implicated in macrophage cholesterol homeostasis were determined by quantitative polymerase chain reaction, and immunoblotting, relative to the housekeeping enzyme, Gapdh, and combined with studies of this molecule on cholesterol esterification, de novo lipid biosynthesis, and induction of apoptosis. Significant differences were determined using analysis of variance, and Dunnett’s or Bonferroni post t-tests, as appropriate. Results The positive control, resveratrol (24 h, significantly enhanced cholesterol efflux to apoA-I at concentrations ≥30 μM. By contrast, cholesterol efflux to apoA-I was significantly inhibited by nigericin (45%; ppAbca1 mRNA. Oligomycin treatment did not affect cholesterol biosynthesis, but significantly inhibited cholesterol esterification following exposure to acetylated LDL, and induced apoptosis at ≥30 μM. Finally, oligomycin induced the expression of genes implicated in both cholesterol efflux (Abca1, Abcg4, Stard1 and cholesterol biosynthesis (Hmgr, Mvk, Scap, Srebf2, indicating profound dysregulation of cholesterol homeostasis. Conclusions Acute loss of mitochondrial function, and in particular Δψm, reduces

  15. Polymeric proanthocyanidins from Sicilian pistachio (Pistacia vera L.) nut extract inhibit lipopolysaccharide-induced inflammatory response in RAW 264.7 cells.

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    Gentile, C; Allegra, M; Angileri, F; Pintaudi, A M; Livrea, M A; Tesoriere, L

    2012-04-01

    Positive effects of pistachio nut consumption on plasma inflammatory biomarkers have been described; however, little is known about molecular events associated with these effects. We studied the anti-inflammatory activity of a hydrophilic extract from Sicilian Pistacia L. (HPE) in a macrophage model and investigated bioactive components relevant to the observed effects. HPE oligomer/polymer proanthocyanidin fractions were isolated by adsorbance chromatography, and components quantified as anthocyanidins after acidic hydrolysis. Isoflavones were measured by gradient elution HPLC analysis. RAW 264.7 murine macrophages were pre-incubated with either HPE (1- to 20-mg fresh nut equivalents) or its isolated components for 1 h, then washed before stimulating with lipopolysaccharide (LPS) for 24 h. Cell viability and parameters associated with Nuclear Factor-κB (NF-κB) activation were assayed according to established methods including ELISA, Western blot, or cytofluorimetric analysis. HPE suppressed nitric oxide (NO) and tumor necrosis factor-α (TNF-α) production and inducible NO-synthase levels dose dependently, whereas inhibited prostaglandin E2 (PGE2) release and decreased cyclo-oxygenase-2 content, the lower the HPE amount the higher the effect. Cytotoxic effects were not observed. HPE also caused a dose-dependent decrease in intracellular reactive oxygen species and interfered with the NF-κB activation. Polymeric proanthocyanidins, but not isoflavones, at a concentration comparable with their content in HPE, inhibited NO, PGE2, and TNF-α formation, as well as activation of IκB-α. Oligomeric proanthocyanidins showed only minor effects. Our results provide molecular evidence of anti-inflammatory activity of pistachio nut and indicate polymeric proanthocyanidins as the bioactive components. The mechanism may involve the redox-sensitive transcription factor NF-κB. Potential effects associated with pistachio nut consumption are discussed in terms of the

  16. Taraxacum officinale Weber extracts inhibit LPS-induced oxidative stress and nitric oxide production via the NF-κB modulation in RAW 264.7 cells.

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    Park, Chung Mu; Park, Ji Young; Noh, Kyung Hee; Shin, Jin Hyuk; Song, Young Sun

    2011-01-27

    The common dandelion (Taraxacum officinale G.H. Weber ex Wiggers, Asteraceae) has been widely used in folklore medicine to treat dyspepsia, heartburn, and spleen and liver disorders. To compare the antioxidative and anti-inflammatory activities of Taraxacum officinale methanol extract (TOME) and water extract (TOWE) in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells and assess their constitutional differences, including luteolin, chicoric acid, and total phenol content. Antioxidative enzyme activities, nitric oxide (NO) production, and inducible NO synthase (iNOS) and nuclear factor (NF)-κB expression were estimated by biochemical analysis, the Griess reaction, reverse transcription-polymerase chain reaction, western hybridization, and electrophoretic mobility shift assay. High-performance liquid chromatography and the Folin-Ciocalteau method were used to analyze functional phytochemicals and total phenol content. TOME and TOWE significantly reduced NO production with an IC(50) of 79.9 and 157.5 μg/mL, respectively, without cytotoxicity. Depleted glutathione (GSH) and antioxidative enzyme activities, including superoxide dismutase, catalase, GSH-peroxidase, and GSH-reductase, were restored by dandelion extracts. Both extracts inhibited LPS-stimulated iNOS gene expression and that of its transcription factor, NF-κB, in parallel with nitrite reduction. TOME showed more potent antioxidative and anti-inflammatory capacities than TOWE, which was attributable to its high total phenol, luteolin, and chicoric acid content. These results indicate that TOME and TOWE inhibit oxidative stress and inflammatory responses through elevated de novo synthesis of antioxidative enzymes and suppression of iNOS expression by NF-κB inactivation. Copyright © 2010 Elsevier Ireland Ltd. All rights reserved.

  17. Cannabinoid CB2 receptors are involved in the protection of RAW264.7 macrophages against the oxidative stress: an in vitro study

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    Sabrina Giacoppo

    2017-01-01

    Full Text Available Research in the last decades has widely investigated the anti-oxidant properties of natural products as a therapeutic approach for the prevention and the treatment of oxidative-stress related disorders. In this context, several studies were aimed to evaluate the therapeutic potential of phytocannabinoids, the bioactive compounds of Cannabis sativa. Here, we examined the anti-oxidant ability of Cannabigerol (CBG, a non-psychotropic cannabinoid, still little known, into counteracting the hydrogen peroxide (H2O2-induced oxidative stress in murine RAW264.7 macrophages. In addition, we tested selective receptor antagonists for cannabinoid receptors and specifically CB1R (SR141716A and CB2R (AM630 in order to investigate through which CBG may exert its action. Taken together, our in vitro results showed that CBG is able to counteract oxidative stress by activation of CB2 receptors. CB2 antagonist pre-treatment indeed blocked the protective effects of CBG in H2O2 stimulated macrophages, while CB1R was not involved. Specifically, CBG exhibited a potent action in inhibiting oxidative stress, by down-regulation of the main oxidative markers (iNOS, nitrotyrosine and PARP-1, by preventing IκB-α phosphorylation and translocation of the nuclear factor-κB (NF-κB and also via the modulation of MAP kinases pathway. On the other hand, CBG was found to increase anti-oxidant defense of cells by modulating superoxide dismutase-1 (SOD-1 expression and thus inhibiting cell death (results focused on balance between Bax and Bcl-2. Based on its antioxidant activities, CBG may hold great promise as an anti-oxidant agent and therefore used in clinical practice as a new approach in oxidative-stress related disorders.

  18. Inhibition of Nitric Oxide (NO Production in Lipopolysaccharide (LPS-Activated Murine Macrophage RAW 264.7 Cells by the Norsesterterpene Peroxide, Epimuqubilin A

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    Leng Chee Chang

    2010-03-01

    Full Text Available Seven norsesterterpene peroxides: epimuqubilin A (1, muqubilone B (2, unnamed cyclic peroxide ester (3, epimuqubilin B (4, sigmosceptrellin A methyl ester (5, sigmosceptrellin A (6, and sigmosceptrellin B methyl ester (7, isolated from the marine sponge Latrunculia sp., were examined with regard to their effects on nitric oxide (NO production in lipopolysaccharide (LPS-activated murine macrophage RAW 264.7 cells. The results indicated epimuqubilin A (1 possessed potent NO inhibitory activity against lipopolysaccharide (LPS-induced nitric oxide release with an IC50 value of 7.4 µM, a level three times greater than the positive control, L-NG-monomethyl arginine citrate, followed by 6 (sigmosceptrellin A, IC50 = 9.9 µM, whereas other compounds exhibited only modest activity (Table 1. These compounds did not show appreciable cytotoxicity at their IC50 values for NO–inhibitory activity. The structure–activity upon NO inhibition could be summarized as follows: (1 a monocyclic carbon skeleton framework was essential for activity,(2 free acids gave higher activity, (3 the orientation of H3-22 with an equatorial position increased activity, and (4 a bicyclic structure reduced activity. This is the first report of a norsesterterpene peroxide with NO–inhibitory activity. In addition, compounds 1–7 were also evaluated for their inhibitory activities in the yeast glycogen synthase kinase-3βassay. In summary, several norsesterterpene peroxides showed novel biological activities of inhibition in NO production, suggesting that these might provide leads for anti-inflammatory or cancer chemopreventive agents.

  19. Adiponectin Inhibits LPS-Induced HMGB1 Release through an AMP Kinase and Heme Oxygenase-1-Dependent Pathway in RAW 264 Macrophage Cells

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    Mohamed Elfeky

    2016-01-01

    Full Text Available High mobility group protein B1 (HMGB1 is a late inflammatory mediator that exaggerates septic symptoms. Adiponectin, an adipokine, has potent anti-inflammatory properties. However, possible effects of adiponectin on lipopolysaccharide- (LPS- induced HMGB1 release are unknown. The aim of this study was to investigate effects of full length adiponectin on HMGB1 release in LPS-stimulated RAW 264 macrophage cells. Treatment of the cells with LPS alone significantly induced HMGB1 release associated with HMGB1 translocation from the nucleus to the cytosol. However, prior treatment with adiponectin suppressed LPS-induced HMGB1 release and translocation. The anti-inflammatory cytokine interleukin- (IL- 10 similarly suppressed LPS-induced HMGB1 release. Adiponectin treatment decreased toll-like receptor 4 (TLR4 mRNA expression and increased heme oxygenase- (HO- 1 mRNA expression without inducing IL-10 mRNA, while IL-10 treatment decreased TLR2 and HMGB1 mRNA expression and increased the expression of IL-10 and HO-1 mRNA. Treatment with the HO-1 inhibitor ZnPP completely prevented the suppression of HMGB1 release by adiponectin but only partially inhibited that induced by IL-10. Treatment with compound C, an AMP kinase (AMPK inhibitor, abolished the increase in HO-1 expression and the suppression of HMGB1 release mediated by adiponectin. In conclusion, our results indicate that adiponectin suppresses HMGB1 release by LPS through an AMPK-mediated and HO-1-dependent IL-10-independent pathway.

  20. A superoxide dismutase purified from the rhizome of Curcuma aeruginosa Roxb. as inhibitor of nitric oxide production in the macrophage-like RAW 264.7 cell line.

    Science.gov (United States)

    Moon-ai, Wanwisa; Niyomploy, Ploypat; Boonsombat, Ruethairat; Sangvanich, Polkit; Karnchanatat, Aphichart

    2012-04-01

    Superoxide dismutase (SOD, EC 1.15.1.1) is a metalloenzyme or antioxidant enzyme that catalyzes the disproportionation of the harmful superoxide anionic radical to hydrogen peroxide and molecular oxygen. Due to its antioxidative effects, SOD has long been applied in medicinal treatment, cosmetic, and other chemical industries. Fifteen Zingiberaceae plants were tested for SOD activity in their rhizome extracts. The crude homogenate and ammonium sulfate cut fraction of Curcuma aeruginosa were found to contain a significant level of SOD activity. The SOD enzyme was enriched 16.7-fold by sequential ammonium sulfate precipitation, diethylaminoethyl cellulose ion exchange, and Superdex 75 gel filtration column chromatography. An overall SOD yield of 2.51 % with a specific activity of 812.20 U/mg was obtained. The enriched SOD had an apparent MW of 31.5 kDa, as judged by sodium dodecyl sulfate polyacrylamide gel electrophoresis, and a pH and temperature optima of 4.0 and 50 °C. With nitroblue tetrazolium and riboflavin as substrates, the K(m) values were 57.31 ± 0.012 and 1.51 ± 0.014 M, respectively, with corresponding V(max) values of 333.7 ± 0.034 and 254.1 ± 0.022 μmol min(-1) mg protein(-1). This SOD likely belongs to the Fe- or Mn-SOD category due to the fact that it was insensitive to potassium cyanide or hydrogen peroxide inhibition, but was potentially weakly stimulated by hydrogen peroxide, and stimulated by Mn(2+)and Fe(2+) ions. Moreover, this purified SOD also exhibited inhibitory effects on lipopolysaccharide-induced nitric oxide production in cultured mouse macrophage cell RAW 264.7 in a dose-dependent manner (IC(50) = 14.36 ± 0.15 μg protein/ml).

  1. Moringa oleifera Flower Extract Suppresses the Activation of Inflammatory Mediators in Lipopolysaccharide-Stimulated RAW 264.7 Macrophages via NF-κB Pathway

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    Woan Sean Tan

    2015-01-01

    Full Text Available Aim of Study. Moringa oleifera Lam. (M. oleifera possess highest concentration of antioxidant bioactive compounds and is anticipated to be used as an alternative medicine for inflammation. In the present study, we investigated the anti-inflammatory activity of 80% hydroethanolic extract of M. oleifera flower on proinflammatory mediators and cytokines produced in lipopolysaccharide- (LPS- induced RAW 264.7 macrophages. Materials and Methods. Cell cytotoxicity was conducted by 3-(4,5-dimethylthiazol-2-yl-2,5-diphenyltetrazolium bromide (MTT assay. Nitric oxide (NO production was quantified through Griess reaction while proinflammatory cytokines and other key inflammatory markers were assessed through enzyme-linked immunosorbent assay (ELISA and immunoblotting. Results. Hydroethanolic extract of M. oleifera flower significantly suppressed the secretion and expression of NO, prostaglandin E2 (PGE2, interleukin- (IL- 6, IL-1β, tumor necrosis factor-alpha (TNF-α, nuclear factor-kappa B (NF-κB, inducible NO synthase (iNOS, and cyclooxygenase-2 (COX-2. However, it significantly increased the production of IL-10 and IκB-α (inhibitor of κB in a concentration dependent manner (100 μg/mL and 200 μg/mL. Conclusion. These results suggest that 80% hydroethanolic extract of M. oleifera flower has anti-inflammatory action related to its inhibition of NO, PGE2, proinflammatory cytokines, and inflammatory mediator’s production in LPS-stimulated macrophages through preventing degradation of IκB-α in NF-κB signaling pathway.

  2. Induction of cytotoxicity and production of inflammatory mediators in raw264.7 macrophages by spores grown on six different plasterboards.

    Science.gov (United States)

    Murtoniemi, T; Nevalainen, A; Suutari, M; Toivola, M; Komulainen, H; Hirvonen, M R

    2001-03-01

    Dampness and microbial growth in buildings are associated with respiratory symptoms in the occupants, but details of the phenomenon are not sufficiently understood. The current study examined the effects of growth conditions provided by six plasterboards on cytotoxicity and inflammatory potential of the spores of Streptomyces californicus, Penicillium spinulosum, Aspergillus versicolor, and Stachybotrys chartarum. The microbes were isolated from mold problem buildings and thereafter grown on six different plasterboards. The spores were harvested, applied to RAW264.7 macrophages (10(4), 10(5), 10(6) spores/10(6) cells), and evaluated 24 h after exposure for the ability to cause cytotoxicity and to stimulate production of nitric oxide (NO), interleukin-1 beta (IL-1beta), tumor necrosis factor alpha (TNFalpha) and interleukin-6 (IL-6). The data indicate clear differences between spores of different microbes in their ability to induce the production of these inflammatory mediators and to cause cell death in macrophages. Also, for each microbe, the induction ability specifically depended on the brand of plasterboard. The spores of Streptomyces californicus collected from all plasterboards were the most potent at inducing NO and cytokine production. Cytotoxicity caused by P. spinulosum and Streptomyces californicus spores was consistent with NO, IL-1beta and IL-6 production induced by those microbes. However, the production of these inflammatory mediators by the spores of Stachybotrys chartarum was not parallel to their ability to cause cell death. The low productions of NO and cytokines were associated with high cytotoxicity caused by the spores of the A. versicolor. These data suggest that growth condition of microbes on different plasterboards affect the ability of microbial spores to induce inflammatory responses and cytotoxicity in macrophages.

  3. Donepezil, an acetylcholinesterase inhibitor, attenuates LPS-induced inflammatory response in murine macrophage cell line RAW 264.7 through inhibition of nuclear factor kappa B translocation.

    Science.gov (United States)

    Arikawa, Mikihiko; Kakinuma, Yoshihiko; Noguchi, Tatsuya; Todaka, Hiroshi; Sato, Takayuki

    2016-10-15

    We have previously demonstrated that the pharmacotherapy with donepezil, an acetylcholinesterase inhibitor, suppresses cardiac remodeling in a mouse model of ischemic heart failure after myocardial infarction (MI). However, the precise mechanisms of the cardioprotective effect of donepezil have not been completely delineated. Because post-ischemic inflammation is a pathological key event in the cardiac remodeling process following MI, we investigated the hypothesis that donepezil acts as an inhibitor of inflammatory mediators. RAW 264.7 murine macrophage cells were pretreated with donepezil (100µM) prior to a pro-inflammatory stimulation by administration of lipopolysaccharide (LPS, 10ng/ml). Donepezil significantly reduced intra- and extracellular levels of various kinds of inflammatory mediators such as TNF-α, IL-1β, IL-2, IL-6 and IL-18 after the LPS stimulation, and attenuated LPS-induced nuclear translocation of nuclear factor-kappa B (NF-κB). These results indicate that donepezil possesses an anti-inflammatory property. However, the inhibitory effect of donepezil on the macrophage inflammatory responses was never reproduced by ACh, nor was disrupted by ACh receptor blockers. Moreover, other kinds of acetylcholinesterase inhibitors failed to inhibit the inflammatory responses in LPS-stimulated macrophage cells. These results suggest that a cholinergic anti-inflammatory pathway would not be involved in the anti-inflammatory effect of donepezil and that the specific characteristics of donepezil in suppressing the LPS-induced cytokine release and the NF-κB activation would be independent of its acetylcholinesterase inhibition. The present study showed that donepezil exerts an anti-inflammatory effect independently of acetylcholinesterase inhibitory action, thereby donepezil may contribute to cardioprotection during cardiac remodeling process in an ischemic heart failure after MI. Copyright © 2016 Elsevier B.V. All rights reserved.

  4. Evidence of biological activity of Mentha species extracts on apoptotic and autophagic targets on murine RAW264.7 and human U937 monocytic cells.

    Science.gov (United States)

    Brahmi, Fatiha; Hadj-Ahmed, Samia; Zarrouk, Amira; Bezine, Maryem; Nury, Thomas; Madani, Khodir; Chibane, Mohamed; Vejux, Anne; Andreoletti, Pierre; Boulekbache-Makhlouf, Lila; Lizard, Gérard

    2017-12-01

    Mints (Lamiaceae) are used as traditional remedies for the treatment of several diseases. Their extracts are recognized as anti-inflammatory compounds. This study characterized the cytotoxic effects of Mentha spicata L. (MS), Mentha pulegium L. (MP) and Mentha rotundifolia (L). Huds (MR) on macrophage cells (RAW264.7; U937) and determined their impact on apoptosis and autophagy, which can play a role in controlling inflammation. The extracts were prepared in culture medium and tested from 25 to 400 μg/mL after 24-48 h of treatment. To show the effect of the aqueous ethanol (50%) extracts on apoptosis and authophagy, the presence of cleaved caspase-3, and the conversion of LC3-I to LC3-II was evaluated by Western blotting. Compared with the MTT assay, crystal violet showed a pronounced decrease in the number of cells with all extracts at 48 h. Calculated IC 50 values were 257.31, 207.82 and 368.02 μg/mL for MS, MP and MR, respectively. A significant increase in PI positive cells was observed with all extracts at 200-400 μg/mL. Mitochondrial dysfunctions and nuclear morphological changes were detected with MS and MR extracts at 400 μg/mL. At this concentration, no cleaved caspase-3 was found whereas stabilized caspase-3 in its dimeric form was identified. MS and MR extracts also favour LC3-I to LC3-II conversion which is a criterion of autophagy. The cytotoxic profiles depend on the extracts considered; MS extract showed the strong activity. However, all the mint extracts studied interact with the apoptotic and autophagic pathways at elevated concentrations.

  5. Effects of Graphene Oxide Nanoparticles on the Immune System Biomarkers Produced by RAW 264.7 and Human Whole Blood Cell Cultures

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    Kim Lategan

    2018-02-01

    Full Text Available Graphene oxide nanoparticles (GONPs have attracted a lot of attention due to their many applications. These applications include batteries, super capacitors, drug delivery and biosensing. However, few studies have investigated the effects of these nanoparticles on the immune system. In this study, the in vitro effects of GONPs on the immune system was evaluated by exposing murine macrophages, RAW 264.7 cells and human whole blood cell cultures (to GONPs. The effects of GONPs on RAW cells were monitored under basal conditions. The whole blood cell cultures were exposed to GONPs in the presence or absence of the mitogens lipopolysaccharide (LPS and phytohaemmagglutinin (PHA. A number of parameters were monitored for both RAW and whole blood cell cultures, these included cytotoxicity, inflammatory biomarkers, cytokines of the acquired immune system and a proteome profile analysis. The GONPs were cytotoxic to both RAW and whole blood cell cultures at 500 μg/mL. In the absence of LPS, GONPs elicited an inflammatory response from the murine macrophage, RAW and whole blood cell cultures at 15.6 and 5 μg/mL respectively. This activation was further corroborated by proteome profile analysis of both experimental cultures. GONPs inhibited LPS induced interleukin 6 (IL-6 synthesis and PHA induced interferon gamma (IFNγ synthesis by whole blood cell cultures in a dose dependent manner. In the absence of mitogens, GONPs stimulated IL-10 synthesis by whole blood cell cultures. The current study shows that GONPs modulate immune system biomarkers and that these may pose a health risk to individuals exposed to this type of nanoparticle.

  6. Inflammation-associated gene expression in RAW 264.7 macrophages induced by toxins from fungi common on damp building materials.

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    Rand, Thomas G; Chang, Carolyn T; McMullin, David R; Miller, J David

    2017-09-01

    Most fungi that grow on damp building materials produce low molecular weight compounds, some of which are known to be toxic. In this study, we tested the hypothesis that exposure to some metabolites of fungi common on damp building materials would result in time-, dose-, and compound-specific responses in the production of various chemokines by RAW 264.7 cells. Cell cultures were exposed to a 10 -7 M or 10 -8 M metabolite dose for 2, 4, 8 or 24h. Metabolite concentrations used were based on those that might be expected in alveolar macrophages due to inhalation exposure from living or working in a damp building. Compared to controls, exposure provoked significant time-, dose- and compound-specific responses manifest as differentially elevated secretion of three of nine cytokines tested in culture supernatant of treated cells. The greatest number of cytokines produced in response to the metabolites tested were in andrastin A-treated cells (GM-CSF, TGFβ1, Tnf-α) followed by koninginin A (TGFβ1 and Tnf-α) and phomenone (GM-CSF, TGFβ1). Chaetoglobosin A, chaetomugilin D and walleminone exposures each resulted in significant time-specific production of Tnf-α only. This investigation adds to a body of evidence supporting the role of low molecular weight compounds from damp building materials as pathogen associated molecular patterns (PAMPs). Along with fungal glucan and chitin, these compounds contribute to the non-allergy based respiratory outcomes for people living and working in damp buildings. Copyright © 2017 Elsevier Ltd. All rights reserved.

  7. Effects of solubility of urban air fine and coarse particles on cytotoxic and inflammatory responses in RAW 264.7 macrophage cell line

    International Nuclear Information System (INIS)

    Jalava, Pasi I.; Salonen, Raimo O.; Pennanen, Arto S.; Happo, Mikko S.; Penttinen, Piia; Haelinen, Arja I.; Sillanpaeae, Markus; Hillamo, Risto; Hirvonen, Maija-Riitta

    2008-01-01

    We investigated the inflammatory and cytotoxic activities of the water-soluble and -insoluble as well as organic-solvent-soluble and -insoluble fractions of urban air fine (PM 2.5-0.2 ) and coarse (PM 10-2.5 ) particulate samples. The samples were collected with a high volume cascade impactor (HVCI) in 7-week sampling campaigns of selected seasons in six European cities. Mouse macrophage cells (RAW 264.7) were exposed to the samples for 24 h. The production of nitric oxide (NO) and proinflammatory cytokines (TNFα, IL-6), and cytotoxicity (MTT-test, apoptosis, cell cycle) were measured. The inflammatory and cytotoxic responses in both size ranges were mostly associated with the insoluble particulate fractions. However, both the water- and organic-solvent-soluble particulate fractions induced TNFα production and apoptosis and had some other cytotoxic effects. Soil-derived water-soluble and -insoluble components of the chemical PM 2.5-0.2 mass closure had consistent positive correlations with the responses, while the correlations were negative with the secondary inorganic anions (NO 3 - , NH 4 + , non-sea-salt SO 4 2- ) and particulate organic matter (POM). With the PM 10-2.5 samples, sea salt and soluble soil components correlated positively with the induced toxic responses. In this size range, a possible underestimation of the insoluble, soil-related compounds containing Si and Ca, and biological components of POM, increased uncertainties in the evaluation of associations of the mass closure components with the responses. It is concluded that insoluble components of the complex urban air particulate mixture exert the highest inflammatory and cytotoxic activities in the macrophage cell line but, at the same time, they may operate as carriers for active water- and lipid-soluble components

  8. Morphological and functional changes in RAW264 macrophage-like cells in response to a hydrated layer of carbonate-substituted hydroxyapatite.

    Science.gov (United States)

    Igeta, Kazuki; Kuwamura, Yuta; Horiuchi, Naohiro; Nozaki, Kosuke; Shiraishi, Daichi; Aizawa, Mamoru; Hashimoto, Kazuaki; Yamashita, Kimihiro; Nagai, Akiko

    2017-04-01

    Synthetic hydroxyapatite (HAp) is used clinically as a material for bone prostheses owing to its good bone-bonding ability; however, it does not contribute to bone remodeling. Carbonate-substituted hydroxyapatite (CAp) has greater bioresorption capacity than HAp while having similar bone-bonding potential, and is therefore considered as a next promising material for bone prostheses. However, the effects of the CAp instability on inflammatory and immune responses are unknown in detail. Here, we show that the surface layer of CAp is more hydrated than that of HAp and induces changes in the shape and function of macrophage-like cells. HAp and CAp were synthesized by wet method and molded into disks. The carbonate content of CAp disks was 6.2% as determined by Fourier transform (FT) infrared spectral analysis. Diffuse reflectance infrared FT analysis confirmed that physisorbed water and surface hydroxyl groups (OH - ) were increased whereas structural OH - was decreased on the CAp as compared to the HAp surface. The degree of hydroxylation in CAp was comparable to that in bone-apatite structures, and the CAp surface exhibited greater hydrophilicity and solubility than HAp. We investigated immune responses to these materials by culturing RAW264 cells (macrophage precursors) on their surfaces. Cell spreading on the CAp disk was suppressed and the secretion level of inflammatory cytokines was reduced as compared to cells grown on HAp. These results indicate that the greater surface hydration of CAp surface can attenuate adverse inflammatory responses to implanted bone prostheses composed of this material. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 1063-1070, 2017. © 2017 Wiley Periodicals, Inc.

  9. A pterostilbene derivative suppresses osteoclastogenesis by regulating RANKL-mediated NFκB and MAPK signaling in RAW264.7 cells.

    Science.gov (United States)

    Nikhil, Kumar; Sharan, Shruti; Roy, Partha

    2015-12-01

    A dysfunctional osteoclast activity is often the cause of bone destructive diseases, such as osteoporosis, periodontitis, erosive arthritis, and cancer. The NFκB ligand (RANKL) has been identified as a major mediator of bone resorption. Agents that suppress RANKL signaling have the potential to inhibit bone resorption or osteoclastogenesis. The present study aimed to determine the effect of a pterostilbene derivative (PTERC-T) for suppressing RANKL or tumor cells-induced osteoclastogenesis in RAW264.7 murine macrophages. Cytotoxicity was measured by MTT assay and inhibitory effect on osteoclastogenesis was analyzed by counting the number of tartrate-resistant acid phosphatase (TRAP)-positive multinucleated cells and measuring the expression levels of the osteoclast-specific genes. The reactive oxygen species (ROS) generation was detected by FACS. Further, signaling pathways were analyzed by immunofluorescence and immunoblot analyses. PTERC-T suppressed the differentiation of monocytes to osteoclasts in a dose and time-dependent manner. The expression of osteoclast marker genes like TRAP, cathepsin K (CTSK), matrix metalloproteinase 9 (MMP9) and transcription factors c-Fos, and nuclear factor of activated T cells cytoplasmic 1 (NFATc1) were also diminished by PTERC-T. PTERC-T scavenged intracellular ROS generation within osteoclast precursors during RANKL-stimulated osteoclastogenesis. Mechanistically, PTERC-T abrogated the phosphorylation of MAPKs (ERK and JNK) and inhibited RANKL-induced activation of NFκB by suppressing IκBα phosphorylation and preventing NFκB/p65 nuclear translocation. This study thus identifies PTERC-T as an inhibitor of osteoclast formation and provides evidence for its role in preventing osteoporosis and other bone related disorders. However, further studies are needed to establish its efficacy in vivo. Copyright © 2015 Institute of Pharmacology, Polish Academy of Sciences. Published by Elsevier Urban & Partner Sp. z o.o. All rights

  10. Pro-inflammatory responses of RAW264.7 macrophages when treated with ultralow concentrations of silver, titanium dioxide, and zinc oxide nanoparticles

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    Giovanni, Marcella [Department of Chemical and Biomolecular Engineering, National University of Singapore, 4 Engineering Drive 4, Singapore 117585 (Singapore); Yue, Junqi; Zhang, Lifeng [PUB, 40 Scotts Road, Singapore 228231 (Singapore); Xie, Jianping [Department of Chemical and Biomolecular Engineering, National University of Singapore, 4 Engineering Drive 4, Singapore 117585 (Singapore); Ong, Choon Nam [Saw Swee Hock School of Public Health, National University of Singapore, 12 Science Drive 2, Singapore 117549 (Singapore); NUS Environmental Research Institute, National University of Singapore, 5A Engineering Drive 1, Singapore 117411 (Singapore); Leong, David Tai, E-mail: cheltwd@nus.edu.sg [Department of Chemical and Biomolecular Engineering, National University of Singapore, 4 Engineering Drive 4, Singapore 117585 (Singapore)

    2015-10-30

    Highlights: • Ultralow levels of common nanoparticles exist in environment and consumer products. • Common nanoparticles at ultralow levels induce mild pro-inflammation by macrophages. • The nanoparticles are cytotoxic only at high doses. - Abstract: To cellular systems, nanoparticles are considered as foreign particles. Upon particles and cells contact, innate immune system responds by activating the inflammatory pathway. However, excessive inflammation had been linked to various diseases ranging from allergic responses to cancer. Common nanoparticles, namely silver, titanium dioxide, and zinc oxide exist in the environment as well as in consumer products at ultralow level of 10{sup −6}–10{sup −3} μg mL{sup −1}. However, so far the risks of such low NPs concentrations remain unexplored. Therefore, we attempted to screen the pro-inflammatory responses after ultralow concentration treatments of the three nanoparticles on RAW264.7 macrophages, which are a part of the immune system, at both cellular and gene levels. Even though cytotoxicity was only observed at nanoparticles concentrations as high as 10 μg mL{sup −1}, through the level of NF-κB and upregulation of pro-inflammatory genes, we observed activation of the induction of genes encoding pro-inflammatory cytokines starting already at 10{sup −7} μg mL{sup −1}. This calls for more thorough characterization of nanoparticles in the environment as well as in consumer products to ascertain the health and safety of the consumers and living systems in general.

  11. Pro-inflammatory agents LPS and IL-6 protect monocytic cell line RAW264.7 from radiation induced cell death

    International Nuclear Information System (INIS)

    Neeraj Kumari; Rai, Yogesh; Pathak, Richa; Kalra, Namita; Sharma, Anjali; Dwarakanath, B.S.; Bhatt, Anant Narayan

    2014-01-01

    Our earlier studies have shown that increased glycolysis protects cells from radiation induced cell death. Pro-inflammatory molecules like LPS have been shown to reduce radiation induced gastro-intestinal syndrome, while IL-6 protects cardiomyocytes from ischemia induced oxidative stress. Interestingly, both pro-inflammatory molecules, LPS and IL-6 induce glycolysis in cells and mimic the high glycolytic phenotype. Therefore, we hypothesize that LPS and IL-6 can protect the hematopoietic cells from radiation induced cell death by inducing glycolysis. To validate our hypothesis, we investigated the response of RAW264.7 cells stimulated with LPS (10 ng/ml) and IL-6 (1 ng/ml), 2 hours prior to irradiation (2 Gy, gamma rays, 60 Co). Both LPS and IL-6 protected cells from radiation induced growth inhibition with > 50% increase in cell number as compared to radiation alone. Under these conditions, IL-6 showed more than 2 fold increase in glycolysis, measured by lactate production, which correlated with increased cell number. To understand the mechanisms underlying IL-6 induced radio-resistance, we examined the effects of IL-6 on anti-oxidant defence and mitochondrial status in irradiated cells. Cells treated with IL-6 showed nearly 40% reduced levels of radiation induced delayed reactive oxygen species (ROS), measured at 24 hours after exposure using DCFH2-DA. The decrease in ROS was linked to increased mitochondrial membrane potential (MMP), thereby suggesting that IL-6 induced reduction in ROS levels and high MMP protects the cell from radiation induced cell death. Our results show that both mitochondrial uncouplers and pro-inflammatory molecules (LPS and IL-6) lead to similar metabolic shift in the form of increased glycolysis leading to enhanced radio-resistance. Therefore, we propose that stimulation of glycolysis can be an useful radioprotective strategy, irrespective of the nature of stimulants. Further studies to understand mechanisms underlying IL-6 induced

  12. Inhibition of inducible nitric oxide synthesis by Cimicifuga racemosa (Actaea racemosa, black cohosh) extracts in LPS-stimulated RAW 264.7 macrophages.

    Science.gov (United States)

    Schmid, Diethart; Gruber, Miriam; Woehs, Florian; Prinz, Sonja; Etzlstorfer, Barbara; Prucker, Christina; Fuzzati, Nicola; Kopp, Brigitte; Moeslinger, Thomas

    2009-08-01

    Cimicifuga racemosa (Actaea racemosa, black cohosh) is used as an anti-inflammatory, antipyretic and analgesic remedy in traditional medicines. The present study focuses on the effects of C. racemosa root extracts on inducible nitric oxide synthase (iNOS) in lipopolysaccharide-stimulated murine macrophages (RAW 264.7). C. racemosa rhizome and phosphate-buffered saline extracts were analysed for phenolcarboxylic acids and triterpene glycosides using an HPLC photodiode array/evaporative light-scattering detector system. iNOS was characterised by measurement of iNOS protein (immunoblotting), iNOS mRNA (semiquantitative competitive RT-PCR), nitric oxide production (nitrite levels) and nuclear translocation of nuclear factor-kappaB (p65 subunit) protein. Incubation of lipopolysaccharide-stimulated macrophages with aqueous C. racemosa extracts (0-6 mg/ml) inhibited nitrite accumulation in a concentration-dependent manner. C. racemosa extracts also reduced iNOS protein expression and iNOS mRNA levels in a dose-dependent manner. C. racemosa extracts did not significantly inhibit iNOS activity and did not affect nuclear translocation of nuclear factor-kappaB (p65 subunit) protein. Incubation with the extract was associated with a concentration-dependent reduction of interferon beta and interferon regulatory factor 1 mRNA. Among the triterpene glycosides, 23-epi-26-deoxyactein was identified as an active principle in C. racemosa extracts. Extracts from the roots of C. racemosa inhibit nitric oxide production by reducing iNOS expression without affecting activity of the enzyme. This might contribute to the anti-inflammatory activities of C. racemosa.

  13. Bioassay guided fractionation and identification of active anti-inflammatory constituent from Delonix elata flowers using RAW 264.7 cells.

    Science.gov (United States)

    Saravanan, S; Hairul Islam, V I; David, H A; Lakshmi Sundaram, R; Chellappandian, M; Balakrishna, K; Rajendran, R; Vijayaraghavan, P; Gabriel Paulraj, M; Ignacimuthu, S

    2015-02-01

    Delonix elata (L.) Gamble (Fabaceae) has been used in the Indian traditional medicine system to treat rheumatism and inflammation. To assess the anti-inflammatory effect of Delonix elata flowers and to isolate the active principle. The prompt anti-inflammatory constituent was isolated from Delonix elata flower extracts using bioassay guided fractionation in liposaccharide (LPS) stimulated RAW 264.7 macrophage cell line. The anti-inflammatory activity of extracts/fractions/sub-fractions/compounds (10, 25, and 50 µg/ml) was evaluated by estimating the levels of nitric oxide (NO), TNF-α, and IL-1β after 24 h of LPS induction (1 μg/ml). The isolated active compound was subjected to NMR, IR, and UV analyses for structure determination. In an attempt to search for anti-inflammatory constituents, the active pure principle was isolated and crystallized as a white compound from Delonix elata flowers methanol extract. This active compound (50 µg/ml) decreased the release of inflammatory mediators levels such as NO (0.263 ± 0.03 µM), TNFα (160.20 ± 17.57 pg/ml), and IL-1β (285.79 ± 15.16 pg/ml) significantly (p UV spectroscopy data. This is the first report of this compound from Delonix elata flowers. The findings of the study support the traditional use of Delonix elata flowers to treat inflammation.

  14. 264 - 267_Uduma_thorium

    African Journals Online (AJOL)

    pc

    2017-06-01

    Jun 1, 2017 ... range is even wider, from Berllium to uranium (Be to. U). The elemental concentration detection ... toys for lead (Pb) content, sorting scrap metals and measuring the lead content of residential paint. They ... associated with metal ore mining and processing in. Nigeria and provides a robust tool for economic.

  15. NF-κB pathway inhibition by anthrocyclic glycoside aloin is key event in preventing osteoclastogenesis in RAW264.7 cells.

    Science.gov (United States)

    Pengjam, Yutthana; Madhyastha, Harishkumar; Madhyastha, Radha; Yamaguchi, Yuya; Nakajima, Yuichi; Maruyama, Masugi

    2016-04-15

    Osteoporosis is a bone pathology leading to increased fracture risk and challenging the quality of life. As current treatments can exhibit deleterious side effects, the use of phyto-compounds with therapeutic and preventive activities against orthopaedic related problems represents a promising alternative. We investigated the effect of aloin, an anthrocyclic compound, on inhibition of osteoclastogenesis using receptor of the nuclear factor κB (NF-κB) ligand (RANKL)-induced RAW264.7 macrophage cells. The inhibitory effect of aloin on in vitro osteoclastogenesis was evaluated by reduction in tartrate-resistant acid phosphatase (TRAP) content and expression levels of osteoclast-specific gene, cathepsin K. Multinuclear formation of osteoclast was assessed with haematoxylin and eosin staining. F4/80 content the marker of the murine monocyte/macrophage cells, was evaluated by immunocytochemistry. The underlining mechanisms were assessed by Western blots and EMSA. Effect of aloin on generation of intracellular reactive oxygen species (ROS) was estimated by dichlorofluorescein diacetate (DCFH-DA). Bone degradation effect was evaluated by bone pit assay. The bone pit culture supernatant was studied by Fluorescein assay. We demonstrated that aloin reduced TRAP content and levels of osteoclast-specific gene and protein, cathepsin K. Treatment with aloin (0.75 µM) prevented multinuclear formation (haematoxylin and eosin staining), reduced intracellular TRAP content (TRAP Staining) and increased F4/80 content (F4/80 immunohistochemistry) in RANKL (20 ng/ml) treated RAW cells. Treatment of the RAW cells with aloin suppressed RANKL-induced NF-κB pathway components like IKKα, IKKβ, Phospho.IKK α/β, NF-κB-p65, Phospho NF-κB-p65 and IκBα. EMSA studies showed aloin dose dependently reduced DNA binding activity of NF-κB. Additionally, in vitro bone pit assay revealed that aloin prevented bone degradation and also decreased the fluorescence content in cells, thus

  16. Evaluation of the Combined Effect of Recombinant High-Density Lipoprotein Carrier and the Encapsulated Lovastatin in RAW264.7 Macrophage Cells Based on the Median-Effect Principle.

    Science.gov (United States)

    Jiang, Cuiping; Zhao, Yi; Yang, Yun; He, Jianhua; Zhang, Wenli; Liu, Jianping

    2018-03-05

    Recombinant high-density lipoprotein (rHDL) displays a similar anti-atherosclerotic effect with native HDL and could also be served as a carrier of cardiovascular drug for atherosclerotic plaque targeting. In our previous studies, rHDL has shown a more potent anti-atherosclerotic efficacy as compared to the other conventional nanoparticles with a payload of lovastatin (LS). Therefore, we hypothesized that a synergistic anti-atherosclerotic effect of the rHDL carrier and the encapsulated LS might exist. In this study, the dose-effect relationships and the combined effect of the rHDL and LS were quantitatively evaluated in RAW 264.7 macrophage cells using the median-effect analysis, in which the rHDL carrier was regarded as a drug combined. Median-effect analysis suggested that rHDL and LS exerted a desirable synergistic inhibition on the oxLDL internalization at a ratio of 6:1 ( D m,LS : D m,rHDL ) in RAW 264.7 macrophage cells. About 50% of the reduction on the intracellular lipid contents was found when RAW264.7 cells were treated with LS-loaded rHDLs at their respective median-effect dose ( D m ) concentrations and a synergistic effect on the mediating cholesterol efflux was also observed, which verified the accuracy of the results obtained from the median-effect analysis. The mechanism underlying the synergistic effect of the rHDL carrier and the drug might be attributed to their potent inhibitory effects on SR-A expression. In conclusion, the median-effect analysis was proven to be a feasible method to quantitatively evaluate the synergistic effect of the biofunctional carrier and the drug encapsulated.

  17. The anti-inflammatory activities of Ainsliaea fragrans Champ. extract and its components in lipopolysaccharide-stimulated RAW264.7 macrophages through inhibition of NF-κB pathway.

    Science.gov (United States)

    Chen, Xin; Miao, Jingshan; Wang, Hao; Zhao, Fang; Hu, Jie; Gao, Peng; Wang, Yue; Zhang, Luyong; Yan, Ming

    2015-07-21

    Ainsliaea fragrans Champ. (A. fragrans) is a traditional Chinese herbal that contains components like 3,5-dicaffeoylquinic acid and 4,5-dicaffeoylquinic acid. It exhibits anti-inflammatory activities which has been used for the treatment of gynecological diseases for many years in China. The aims of the present study were to investigate the anti-inflammatory activities of A. fragrans and elucidate the underlying mechanisms with regard to its molecular basis of action for the best component. The anti-inflammatory effects of A. fragrans were studied by using lipopolysaccharide (LPS)-stimulated activation of nitric oxide (NO) in mouse RAW264.7 macrophages. Expression of inducible NO synthase (iNOS) and pro-inflammatory cytokines, inhibitory κBα (IκBα) degradation and nuclear translocation of NF-κB p65 were further investigated. The present study demonstrated that A. fragrans could suppress the production of NO in LPS-stimulated RAW264.7 macrophages. Further investigations showed A. fragrans could suppress iNOS expression. A. fragrans also inhibited the expression of tumor necrosis factor-alpha and interleukin-6. A. fragrans significantly decreased the degradation of IκBα, reduced the level of nuclear translocation of p65. All these results suggested the inhibitory effects of A. fragrans on the production of inflammatory mediators through the inhibition of the NF-κB activation pathway. Our results indicated that A. fragrans inhibited inflammatory events and iNOS expression in LPS-stimulated RAW264.7 cells through the inactivation of NF-κB pathway. This study gives scientific evidence that validate the use of A. fragrans in treatment of patients with gynecological diseases in clinical practice in traditional Chinese medicine. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

  18. n-Butanol extracts of Panax notoginseng suppress LPS-induced MMP-2 expression in periodontal ligament fibroblasts and inhibit osteoclastogenesis by suppressing MAPK in LPS-activated RAW264.7 cells.

    Science.gov (United States)

    Jang, Young-Joo; Kim, Mee-Eun; Ko, Seon-Yle

    2011-11-01

    Periodontitis is a group of inflammatory diseases that affect connective tissue attachments and the supporting bone that surround the teeth. Osteoclasts are responsible for skeletal modeling and remodeling but may also destroy bone in several bone diseases, including osteoporosis and periodontitis. This study examined the anti-inflammatory effects of Panax notoginseng (PN) on periodontal ligament fibroblasts (PDLFs) and RAW264.7 cells under lipopolysaccharide (LPS) induced inflammatory conditions. The effects of PN on PDLFs were determined by measuring the cell viability and mRNA expression of tissue-destructive proteins. The effects of PN on osteoclasts were examined by measuring the following: (1) the cell viability, (2) the formation of Tartrate-resistant acid phosphatase (TRAP)(+) multinucleated cells, (3) MAPK signaling pathways, (4) mRNA expression of inflammatory-related proteins and (5) nitric oxide (NO) production. The n-butanol extracts of PN (bPN) increased the cell proliferation of the PDLFs and decreased the mRNA expression of matrix metalloproteinase (MMP)-2 in the PDLFs. bPN inhibited the formation of LPS-stimulated TRAP(+) multinucleated cells. bPN also inhibited the LPS-stimulated activation of JNK and ERK signaling, and inhibited the LPS-stimulated degradation of I(K)B in the RAW264.7 cells. In addition, bPN decreased the mRNA expression of MMP-9 and iNOS, which are involved in the range of pathophysiological processes, such as inflammation in the RAW264.7 cells. NO production was also decreased via the inhibition of iNOS. These findings suggest that bPN has therapeutic effects on bone-destructive processes, such as those that occur in periodontal diseases. Copyright © 2011 Elsevier Ltd. All rights reserved.

  19. Ergolide, sesquiterpene lactone from Inula britannica, inhibits inducible nitric oxide synthase and cyclo-oxygenase-2 expression in RAW 264.7 macrophages through the inactivation of NF-κB

    Science.gov (United States)

    Whan Han, Jeung; Gon Lee, Byeong; Kee Kim, Yong; Woo Yoon, Jong; Kyoung Jin, Hye; Hong, Sungyoul; Young Lee, Hoi; Ro Lee, Kang; Woo Lee, Hyang

    2001-01-01

    We investigated the mechanism of suppression of inducible nitric oxide synthase (iNOS) and cyclo-oxygenase-2 (COX-2) by ergolide, sesquiterpene lactone from Inula britannica.iNOS activity in cell-free extract of LPS/IFN-γ-stimulated RAW 264.7 macrophages was markedly attenuated by the treatment with ergolide. Its inhibitory effect on iNOS was paralleled by decrease in nitrite accumulation in culture medium of LPS/IFN-γ-stimulated RAW 264.7 macrophages in a concentration-dependent manner. However, its inhibitory effect does not result from direct inhibition of the catalytic activity of NOS.Ergolide markedly decreased the production of prostaglandin E2 (PGE2) in cell-free extract of LPS/IFN-γ-stimulated RAW 264.7 macrophages in a concentration-dependent manner, without alteration of the catalytic activity of COX-2 itself.Ergolide decreased the level of iNOS and COX-2 protein, and iNOS mRNA caused by stimulation of LPS/IFN-γ in a concentration-dependent manner, as measured by Western blot and Northern blot analysis, respectively.Ergolide inhibited nuclear factor-κB (NF-κB) activation, a transcription factor necessary for iNOS and COX-2 expression in response to LPS/IFN-γ. This effect was accompanied by the parallel reduction of nuclear translocation of subunit p65 of NF-κB as well as IκB-α degradation. In addition, these effects were completely blocked by treatment of cysteine, indicating that this inhibitory effect of ergolide could be mediated by alkylation of NF-κB itself or an upstream molecule of NF-κB.Ergolide also directly inhibited the DNA-binding activity of active NF-κB in LPS/IFN-γ-pretreated RAW 264.7 macrophages.These results demonstrate that the suppression of NF-κB activation by ergolide might be attributed to the inhibition of nuclear translocation of NF-κB resulted from blockade of the degradation of IκB and the direct modification of active NF-κB, leading to the suppression of the expression of iNOS and COX-2, which play important

  20. Ergolide, sesquiterpene lactone from Inula britannica, inhibits inducible nitric oxide synthase and cyclo-oxygenase-2 expression in RAW 264.7 macrophages through the inactivation of NF-kappaB.

    Science.gov (United States)

    Whan Han, J; Gon Lee, B; Kee Kim, Y; Woo Yoon, J; Kyoung Jin, H; Hong, S; Young Lee, H; Ro Lee, K; Woo Lee, H

    2001-06-01

    We investigated the mechanism of suppression of inducible nitric oxide synthase (iNOS) and cyclo-oxygenase-2 (COX-2) by ergolide, sesquiterpene lactone from Inula britannica. iNOS activity in cell-free extract of LPS/IFN-gamma-stimulated RAW 264.7 macrophages was markedly attenuated by the treatment with ergolide. Its inhibitory effect on iNOS was paralleled by decrease in nitrite accumulation in culture medium of LPS/IFN-gamma-stimulated RAW 264.7 macrophages in a concentration-dependent manner. However, its inhibitory effect does not result from direct inhibition of the catalytic activity of NOS. Ergolide markedly decreased the production of prostaglandin E(2) (PGE(2)) in cell-free extract of LPS/IFN-gamma-stimulated RAW 264.7 macrophages in a concentration-dependent manner, without alteration of the catalytic activity of COX-2 itself. Ergolide decreased the level of iNOS and COX-2 protein, and iNOS mRNA caused by stimulation of LPS/IFN-gamma in a concentration-dependent manner, as measured by Western blot and Northern blot analysis, respectively. Ergolide inhibited nuclear factor-kappaB (NF-kappaB) activation, a transcription factor necessary for iNOS and COX-2 expression in response to LPS/IFN-gamma. This effect was accompanied by the parallel reduction of nuclear translocation of subunit p65 of NF-kappaB as well as IkappaB-alpha degradation. In addition, these effects were completely blocked by treatment of cysteine, indicating that this inhibitory effect of ergolide could be mediated by alkylation of NF-kappaB itself or an upstream molecule of NF-kappaB. Ergolide also directly inhibited the DNA-binding activity of active NF-kappaB in LPS/IFN-gamma-pretreated RAW 264.7 macrophages. These results demonstrate that the suppression of NF-kappaB activation by ergolide might be attributed to the inhibition of nuclear translocation of NF-kappaB resulted from blockade of the degradation of IkappaB and the direct modification of active NF-kappaB, leading to the

  1. Administration of Protein kinase D1 induce an immunomodulatory effect on lipopolysaccharide-induced intestinal inflammation in a co-culture model of intestinal epithelial Caco-2 cells and RAW 264.7 macrophage cells

    DEFF Research Database (Denmark)

    Nielsen, Ditte Søvsø Gundelund; Fredborg, Marlene; Andersen, Vibeke

    2017-01-01

    the effects of human PKD1 in relation to intestinal inflammation, using a co-culture model of intestinal epithelial Caco-2 cells and RAW264.7 macrophages. An inflammatory response was induced in the macrophages by lipopolysaccharide (LPS), upregulating the expression of tumour necrosis factor alpha (TNF......-α), interleukin- (IL-) 1β, and IL-6 besides increasing the secretion of TNF-α protein. The effect of administering PKD1 to Caco-2 was evaluated in relation to both amelioration of inflammation and the ability to suppress inflammation initiation. Administration of PKD1 (10–100 ng/ml) following induction...

  2. Distribution of CCR5-Delta32, CCR5 promoter 59029 A/G, CCR2-64I and SDF1-3'A genetic polymorphisms in HIV-1 infected and uninfected patients in the west region of Cameroon.

    Science.gov (United States)

    Nkenfou, Céline Nguefeu; Mekue, Linda Chapdeleine Mouafo; Nana, Christelle Tafou; Kuiate, Jules Roger

    2013-07-23

    Genetic variants of the genes encoding human immunodeficiency virus-1 (HIV-1) co-receptors and their ligands, like CC-chemokine receptor 5 delta 32 mutation (CCR5-Delta32), CCR5 promoter A/G (Adenine/Guanine), CC-chemokine receptor 2 mutation 64 isoleucine (CCR2-64I) and the stromal cell-derived factor 3'A mutation (SDF1-3'A), are involved in the susceptibility to HIV-1 infection and progression. The prevalence of these mutations varies by region. However, little is known about their distribution in the population of Dschang, located in the west region of Cameroon. The prevalence of HIV in the west region of Cameroon is lower than elsewhere in Cameroon. The objectives of this study were to determine the distribution of four AIDS Related Gene (ARG) variants in HIV-infected and non-infected population of Cameroon especially in the west region and to estimate the contribution of these variants to the susceptibility or resistance to HIV infection. We also aimed to evaluate the effectiveness of genotyping using dried blood spot (DBS) samples. A total of 179 participants were recruited from two hospitals in Dschang in the west region of Cameroon. Their genotypes for CCR5-Delta32, CCR5 promoter 59029A/G, CCR2-64I and SDF1-3'A were analyzed using polymerase chain reaction (PCR) and restriction fragment length polymorphisms. A total of 179 participants were enrolled in the study. Among them, 32 (17.9%) were HIV positive and 147 (82.1%) were HIV negative. The allelic frequencies of these genes were: 0%, 49.72%, 17.6% and 100% respectively for CCR5-Delta32, CCR5 promoter 59029A/G, CCR2-64I and SDF1-3'A. No individual was found to carry the CCR5-Delta 32 mutation. All participants recruited were heterozygous for the SDF1-3'A allele. Our data suggest that the CCR5-Delta32 cannot account for the protection as it was completely absent in our population. SDF1-3'A variants, may be in association with other polymorphisms, may account for the overall protection from HIV-1 infection

  3. Gelidium elegans, an edible red seaweed, and hesperidin inhibit lipid accumulation and production of reactive oxygen species and reactive nitrogen species in 3T3-L1 and RAW264.7 cells.

    Science.gov (United States)

    Jeon, Hui-Jeon; Seo, Min-Jung; Choi, Hyeon-Son; Lee, Ok-Hwan; Lee, Boo-Yong

    2014-11-01

    Gelidium elegans is an edible red alga native to the intertidal area of northeastern Asia. We investigated the effect of G. elegans extract and its main flavonoids, rutin and hesperidin, on lipid accumulation and the production of reactive oxygen species (ROS) and reactive nitrogen species (RNS) in 3T3-L1 and RAW264.7 cells. Our data show that G. elegans extract decreased lipid accumulation and ROS/RNS production in a dose-dependent manner. The extract also inhibited the mRNA expression of adipogenic transcription factors, such as peroxisome proliferator-activated receptor gamma and CCAAT/enhancer-binding protein alpha, while enhancing the protein expression of the antioxidant enzymes superoxide dismutases 1 and 2, glutathione peroxidase, and glutathione reductase compared with controls. In addition, lipopolysaccharide-induced nitric oxide production was significantly reduced in G. elegans extract-treated RAW264.7 cells. In analysis of the effects of G. elegans flavonoids on lipid accumulation and ROS/RNS production, only hesperidin showed an inhibitory effect on lipid accumulation and ROS production; rutin did not affect adipogenesis and ROS status. The antiadipogenic effect of hesperidin was evidenced by the downregulation of peroxisome proliferator-activated receptor gamma, CCAAT/enhancer-binding protein alpha, and fatty acid binding protein 4 gene expression. Collectively, our data suggest that G. elegans is a potential food source containing antiobesity and antioxidant constituents. Copyright © 2014 John Wiley & Sons, Ltd.

  4. 8-Hydroxyquinoline inhibits iNOS expression and nitric oxide production by down-regulating LPS-induced activity of NF-κB and C/EBPβ in Raw 264.7 cells

    International Nuclear Information System (INIS)

    Kim, Young-Ho; Woo, Kyung Jin; Lim, Jun Hee; Kim, Shin; Lee, Tae Jin; Jung, Eun Mi; Lee, Jin-Man; Park, Jong-Wook; Kwon, Taeg Kyu

    2005-01-01

    In activated macrophage, large amounts of nitric oxide (NO) are generated by inducible nitric oxide synthase (iNOS), resulting in acute or chronic inflammatory disorders. In Raw 264.7 cells stimulated with lipopolysaccharide (LPS) to mimic inflammation, 8-hydroxyquinoline (8HQ) inhibited the LPS-induced expression of both iNOS protein and mRNA in a parallel dose-dependent manner. 8HQ did not enhance the degradation of iNOS mRNA. To investigate the mechanism by which 8HQ inhibits iNOS gene expression, we examined the activation of MAP kinases in Raw 264.7 cells. We did not observe any significant change in the phosphorylation of MAPKs between LPS alone and LPS plus 8HQ-treated cells. Moreover, 8HQ significantly inhibited the DNA-binding activity of nuclear factor-κB (NF-κB) and CCAAT/enhancer-binding protein β (C/EBPβ), but not activator protein-1 and cAMP response element-binding protein. Taken together, these results suggest that 8HQ acts to inhibit inflammation through inhibition of NO production and iNOS expression through blockade of C/EBPβ DNA-binding activity and NF-κB activation

  5. Short-term heating reduces the anti-inflammatory effects of fresh raw garlic extracts on the LPS-induced production of NO and pro-inflammatory cytokines by downregulating allicin activity in RAW 264.7 macrophages.

    Science.gov (United States)

    Shin, Jung-Hye; Ryu, Ji Hyeon; Kang, Min Jung; Hwang, Cho Rong; Han, Jaehee; Kang, Dawon

    2013-08-01

    Garlic has a variety of biologic activities, including anti-inflammatory properties. Although garlic has several biologic activities, some people dislike eating fresh raw garlic because of its strong taste and smell. Therefore, garlic formulations involving heating procedures have been developed. In this study, we investigated whether short-term heating affects the anti-inflammatory properties of garlic. Fresh and heated raw garlic extracts (FRGE and HRGE) were prepared with incubation at 25 °C and 95 °C, respectively, for 2 h. Treatment with FRGE and HRGE significantly reduced the LPS-induced increase in the pro-inflammatory cytokine concentration (TNF-α, IL-1β, and IL-6) and NO through HO-1 upregulation in RAW 264.7 macrophages. The anti-inflammatory effect was greater in FRGE than in HRGE. The allicin concentration was higher in FRGE than in HRGE. Allicin treatment showed reduced production of pro-inflammatory cytokines and NO and increased HO-1 activity. The results show that the decrease in LPS-induced NO and pro-inflammatory cytokines in RAW 264.7 macrophages through HO-1 induction was greater for FRGE compared with HRGE. Additionally, the results indicate that allicin is responsible for the anti-inflammatory effect of FRGE. Our results suggest a potential therapeutic use of allicin in the treatment of chronic inflammatory disease. Copyright © 2013 Elsevier Ltd. All rights reserved.

  6. Anti-Inflammatory Effect of Methylpenicinoline from a Marine Isolate of Penicillium sp. (SF-5995: Inhibition of NF-κB and MAPK Pathways in Lipopolysaccharide-Induced RAW264.7 Macrophages and BV2 Microglia

    Directory of Open Access Journals (Sweden)

    Dong-Cheol Kim

    2014-11-01

    Full Text Available In the course of a search for anti-inflammatory metabolites from marine-derived fungi, methylpenicinoline (1 was isolated from a marine isolate of Penicillin sp. Compound 1 inhibited lipopolysaccharide (LPS-stimulated nitric oxide (NO production by suppressing the expression of inducible NO synthase (iNOS in RAW264.7 macrophages and BV2 microglia. It also attenuated prostaglandin E2 (PGE2 production by suppressing cyclooxygenase-2 (COX-2 expression in a concentration-dependent manner (from 10 μM to 80 μM without affecting cell viability. In addition, compound 1 reduced the production of the pro-inflammatory cytokine interleukin-1β (IL-1β. In a further study designed to elucidate the mechanism of its anti-inflammatory effects, compound 1 was shown to block nuclear factor-kappa B (NF-κB activation in LPS-induced RAW264.7 macrophages and BV2 microglia by inhibiting the phosphorylation of inhibitor kappa B-α (IκB-α, thereby suppressing the nuclear translocation of NF-κB dimers, namely p50 and p65, that are known to be crucial molecules associated with iNOS and COX-2 expression. In addition, compound 1 inhibited the activation of mitogen-activated protein kinase (MAPK pathways. Taken together, the results suggest that compound 1 might be a valuable therapeutic agent for the treatment of anti-inflammatory and anti-neuroinflammatory diseases.

  7. Inhibitory Effects of Chemical Compounds Isolated from the Rhizome of Smilax glabra on Nitric Oxide and Tumor Necrosis Factor-α Production in Lipopolysaccharide-Induced RAW264.7 Cell

    Directory of Open Access Journals (Sweden)

    Chuan-li Lu

    2015-01-01

    Full Text Available The rhizome of Smilax glabra has been used for a long time as both food and folk medicine in many countries. The present study focused on the active constituents from the rhizome of S. glabra, which possess potential anti-inflammatory activities. As a result, nine known compounds were isolated from the rhizome of S. glabra with the bioassay-guiding, and were identified as syringaresinol (1, lasiodiplodin (2, de-O-methyllasiodiplodin (3, syringic acid (4, 1,4-bis(4-hydroxy-3,5-dimethoxyphenyl-2,3-bis(hydroxymethyl-1,4-butanediol (5, lyoniresinol (6, trans-resveratrol (7, trans-caffeic acid methyl ester (8, and dihydrokaempferol (9. Among these compounds, 2 and 3 were isolated for the first time from S. glabra. In addition, the potential anti-inflammatory activities of the isolated compounds were evaluated in vitro in lipopolysaccharide- (LPS- induced RAW264.7 cells. Results indicated that 4 and 7 showed significant inhibitory effects on NO production of RAW264.7 cells, and 1, 2, 3, and 5 showed moderate suppression effects on induced NO production. 1, 7, and 5 exhibited high inhibitory effects on TNF-α production, with the IC50 values less than 2.3, 4.4, and 16.6 μM, respectively. These findings strongly suggest that compounds 1, 2, 3, 4, 5, 7, and 9 were the potential anti-inflammatory active compositions of S. glabra.

  8. Optimization of Astilbin Extraction from the Rhizome of Smilax glabra, and Evaluation of Its Anti-Inflammatory Effect and Probable Underlying Mechanism in Lipopolysaccharide-Induced RAW264.7 Macrophages

    Directory of Open Access Journals (Sweden)

    Chuan-Li Lu

    2015-01-01

    Full Text Available Astilbin, a dihydroflavonol derivative found in many food and medicine plants, exhibited multiple pharmacological functions. In the present study, the ethanol extraction of astilbin from the rhizome of smilax glabra Roxb was optimized by response surface methodology (RSM using Box-Behnken design. Results indicated that the obtained experimental data was well fitted to a second-order polynomial equation by using multiple regression analysis, and the optimal extraction conditions were identified as an extraction time of 40 min, ethanol concentration of 60%, temperature of 73.63 °C, and liquid-solid ratio of 29.89 mL/g for the highest predicted yield of astilbin (15.05 mg/g, which was confirmed through validation experiments. In addition, the anti-inflammatory efficiency of astilbin was evaluated in lipopolysaccharide (LPS-induced RAW 264.7 cells. Results showed that astilbin, at non-cytotoxicity concentrations, significantly suppressed the production of nitric oxide (NO and tumor necrosis factor-α (TNF-α, as well as the mRNA expression of inducible nitric oxide synthase (iNOS and TNF-α in LPS-induced RAW 264.7 cells, but did not affect interleukin-6 (IL-6 release or its mRNA expression. These effects may be related to its up-regulation of the phosphorylation of p65, extracellular signal-regulated kinases 1/2 (ERK1/2 and c-Jun N-terminal kinase (JNK.

  9. Comparative inhibitory effects of magnolol, honokiol, eugenol and bis-eugenol on cyclooxygenase-2 expression and nuclear factor-kappa B activation in RAW264.7 macrophage-like cells stimulated with fimbriae of Porphyromonas gingivalis.

    Science.gov (United States)

    Murakami, Yukio; Kawata, Akifumi; Seki, Yuya; Koh, Teho; Yuhara, Kenji; Maruyama, Takehisa; Machino, Mamoru; Ito, Shigeru; Kadoma, Yoshinori; Fujisawa, Seiichiro

    2012-01-01

    The anti-inflammatory activity of magnolol and related compounds is currently a focus of interest. In the present study, the inhibitory effects of these compounds on cyclooxygenase (COX-2) expression and nuclear factor-kappa B (NF-κB) activation were investigated in RAW264.7 macrophage-like cells stimulated with the fimbriae of Porphyromonas gingivalis, an oral anaerobe. The cytotoxicity of magnolol, honokiol, eugenol and bis-eugenol against RAW264.7 cells was determined using a cell counting kit (CCK-8). The regulatory effect of these compounds on the expression of COX-2 mRNA, stimulated by exposure to the fimbriae was investigated by real-time polymerase chain reaction (PCR). NF-κB activation was evaluated by enzyme-linked immunosorbent assay (ELISA)-like microwell colorimetric transcription factor activity assay (Trans-AM) and western blot analysis. The radical-scavenging activity was determined using the induction period method in the methyl methacrylate-azobisisobutyronitrile (AIBN) polymerization system under nearly anaerobic conditions. The phenolic bond dissociation enthalpy (BDE) and orbital energy were calculated at the density functional theory (DFT) B3LYP/6-31G* level. The cytotoxicity against RAW264.7 cells declined in the order bis-eugenol>eugenol> honokiol>magnolol, whereas the radical-scavenging activity declined in the order honokiol, bis-eugenol>magnolol> eugenol. Magnolol and honokiol significantly inhibited the fimbria-induced expression of COX-2 at non-cytotoxic concentrations. Both the fimbria-stimulated binding of NF-κB to its consensus sequence and phosphorylation-dependent proteolysis of inhibitor κB-α were markedly inhibited by magnilol and honokiol, whereas eugenol and bis-eugenol did not inhibit COX-2 expression and NF-κB activation. Magnolol and honokiol possessed a high electronegativity (χ) value. Magnolol and honokiol exhibit antioxidative activity, low cytotoxicity, and anti-inflammatory activity. These compounds may be

  10. No evidence of a higher 10 year period prevalence of diabetes among 77,885 twins compared with 215,264 singletons from the Danish birth cohorts 1910-1989

    DEFF Research Database (Denmark)

    Petersen, Inge; Nielsen, Morten Frost; Beck-Nielsen, Henning

    2011-01-01

    AIMS/HYPOTHESIS: Previous Danish twin studies have found a highly increased risk of precursors of type 2 diabetes as well as a higher prevalence of type 2 diabetes among twins compared with singletons. Likewise, small-scale studies of Danish twins have shown that monozygotic twins have a higher...... risk of developing precursors of type 2 diabetes compared with dizygotic twins. In the present register-based study, the 10 year period diabetes prevalence in Danish twins is compared with that in a random sample of Danish citizens. Furthermore, the 10 year period prevalence of diabetes in monozygotic...... twins is compared with that in dizygotic twins. METHODS: The study population consisted of twins (n = 77,885) identified in the Danish Twin Registry, and a 5% random sample (n = 215,264) from the birth cohorts 1910-1989. We identified diabetes patients by means of three nationwide Danish health...

  11. Inhibitory Effect of Ferulic Acid and Isoferulic Acid on the Production of Macrophage Inflammatory Protein-2 in Response to Respiratory Syncytial Virus Infection in RAW264.7 Cells

    Directory of Open Access Journals (Sweden)

    S. Sakai

    1999-01-01

    Full Text Available We investigated the effect of ferulic acid (FA and isoferulic acid (IFA, which are the main active components of the rhizoma of Cimicifuga heracleifolia (CH, an anti-inflammatory drug used frequently in Japanese traditional medicine, on the production of macrophage inflammatory protein-2 (MIR-2 in a murine macrophage cell line, RAW264.7, in response to respiratory syncytial virus (RSV infection. Following the exposure of cells to RSV for 20 h, the MIP-2 level in condition medium was increased to about 20 ng/ml, although this level in mock-infected cells was negligible. In the presence of either FA or IFA, RSV-infected cells reduced MIP-2 production in a dose-dependent manner. These data suggest that FA and IFA might be responsible, at least in part, for the anti-inflammatory drug effect of CH extract through the inhibition of MIP-2 production.

  12. ROS-induced toxicity: exposure of 3T3, RAW264.7, and MCF7 cells to superparamagnetic iron oxide nanoparticles results in cell death by mitochondria-dependent apoptosis

    Energy Technology Data Exchange (ETDEWEB)

    Hsieh, Hui-Chen, E-mail: d93548008@ntu.edu.tw; Chen, Chung-Ming, E-mail: chung@ntu.edu.tw [National Taiwan University, Institute of Biomedical Engineering (China); Hsieh, Wen-Yuan, E-mail: hsiehw@itri.org.tw [Industrial Technology Research Institute, Biomedical Technology and Device Research Labs (China); Chen, Ching-Yun, E-mail: chingyun523@gmail.com; Liu, Chia-Ching, E-mail: d95548005@ntu.edu.tw; Lin, Feng-Huei, E-mail: double@ntu.edu.tw [National Taiwan University, Institute of Biomedical Engineering (China)

    2015-02-15

    Superparamagnetic nanoparticles (Fe{sub 3}O{sub 4}, SPIO) have been used as magnetic resonance imaging enhancers for years. However, bio-safety issues concerning nanoparticles remain largely unexplored. Of particular concern is the possible cellular impact of nanoparticles during SPIO uptake and subsequent oxidative stress. SPIO causes cell death by apoptosis via a little understood mitochondrial pathway. To more closely examine this process, three kinds of cells—3T3, RAW264.7, and MCF7—were treated with SPIO coated with polyethylene glycol (SPIO-PEG) and monitored by transmission electron microscopy (TEM), using cytotoxicity evaluation, mitochondrial activity, reactive oxygen species (ROS) generation, and Annexin V assay. TEM revealed that SPIO-PEG nanoparticles surrounded the cellular endosome membrane, creating a bulge in the endosome. Compared to 3T3 cells, greater numbers of SPIO-PEG nanoparticles infiltrated the mitochondria of RAW264.7 and MCF7 cells. SPIO-PEG residency is associated with boosted ROS, with elevated levels of mitochondrial activity, and advancement of cell apoptosis. Furthermore, correlation analysis showed that a polynomial model demonstrates a better fit than a linear model in MCF7, implying that cytotoxicity may have alternative impacts on cell death at different concentrations. Thus, we believe that MCF7 cell death results from the apoptosis pathway triggered by mitochondria, and we find lower cytotoxicity in 3T3. We propose that optimal levels of SPIO-PEG nanoparticles lead to increased levels of ROS and a resulting oxidative stress environment which will kill only cancer cells while sparing normal cells. This finding has great potential for use in cancer therapies in the future.

  13. α-Solanine Isolated From Solanum Tuberosum L. cv Jayoung Abrogates LPS-Induced Inflammatory Responses Via NF-κB Inactivation in RAW 264.7 Macrophages and Endotoxin-Induced Shock Model in Mice.

    Science.gov (United States)

    Shin, Ji-Sun; Lee, Kyoung-Goo; Lee, Hwi-Ho; Lee, Hae Jun; An, Hyo-Jin; Nam, Jung-Hwan; Jang, Dae Sik; Lee, Kyung-Tae

    2016-10-01

    α-Solanine, a trisaccharide glycoalkaloid, has been reported to possess anti-cancer effects. In this study, we investigated the anti-inflammatory effects of α-solanine isolated from "Jayoung" a dark purple-fleshed potato by examining its in vitro inhibitory effects on inducible nitric-oxide synthase (iNOS), cyclooxygenase-2 (COX-2), and pro-inflammatory cytokines in LPS-induced RAW 264.7 macrophages and its in vivo effects on LPS-induced septic shock in a mouse model. α-Solanine suppressed the expression of iNOS and COX-2 both at protein and mRNA levels and consequently inhibited nitric oxide (NO) and prostaglandin E2 (PGE2 ) production in LPS-induced RAW 264.7 macrophages. α-Solanine also reduced the production and mRNA expression of interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), and interleukin-1β (IL-1β) induced by LPS. Furthermore, molecular mechanism studies indicated that α-solanine inhibited LPS-induced activation of nuclear factor-κB (NF-κB) by reducing nuclear translocation of p65, degradation of inhibitory κBα (IκBα), and phosphorylation of IκB kinaseα/β (IKKα/β). In an in vivo experiment of LPS-induced endotoxemia, treatment with α-solanine suppressed mRNA expressions of iNOS, COX-2, IL-6, TNF-α, and IL-1β, and the activation of NF-κB in liver. Importantly, α-solanine increased the survival rate of mice in LPS-induced endotoxemia and polymicrobial sepsis models. Taken together, our data suggest that the α-solanine may be a promising therapeutic against inflammatory diseases by inhibiting the NF-κB signaling pathway. J. Cell. Biochem. 117: 2327-2339, 2016. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

  14. Identification of JAK-STAT Pathways as Important for the Anti-Inflammatory Activity of a Hypericum Perforatum Fraction and Bioactive Constituents in Raw 264.7 Mouse Macrophages

    Science.gov (United States)

    Hammer, Kimberly D. P.; Yum, Man-Yu; Dixon, Philip M.; Birt, Diane F.

    2010-01-01

    Hypericum perforatum extracts have been used to treat diseases including mild-to-moderate depression and inflammatory conditions. It is particularly important to identify which constituents present in the H. perforatum extracts are responsible for its' anti-inflammatory activity since consumers are taking H. perforatum preparations to treat inflammation. We used a combination of four putative bioactive constituents, called the 4-component-system, that interacted synergistically to explain the light-activated anti-inflammatory activity of an H. perforatum fraction in RAW 264.7 mouse macrophages and combined the constituents at concentrations detected in the fraction to identify key molecular targets. LPS was used to model an inflammatory response and the 4-component-system and H. perforatum fraction were used as treatments that inhibited LPS induced prostaglandin E2 (PGE2) production in RAW 264.7 mouse macrophages in studies of gene expression profiles. We used Affymetrix genechips, statistical analysis, and quantitative real-time PCR to identify key gene targets of the 4-component-system and the sub-fraction from an H. perforatum ethanol extract. The H. perforatum sub-fraction with or without LPS stimulation affected far more genes than the 4-component-system with and without LPS. Genes involved in Janus kinase and signal transducer and activator of transcription (JAK-STAT) and eicosanoid pathways were identified that could account for the reduction in PGE2 seen with both treatments in the LPS-stimulated macrophages. Ten genes may be particularly important targets for the activity of the 4-component-system and the fraction with LPS stimulation and these genes were involved in inflammatory signaling pathways, namely JAK-STAT and eicosanoid pathways. PMID:20303133

  15. ROS-induced toxicity: exposure of 3T3, RAW264.7, and MCF7 cells to superparamagnetic iron oxide nanoparticles results in cell death by mitochondria-dependent apoptosis

    Science.gov (United States)

    Hsieh, Hui-Chen; Chen, Chung-Ming; Hsieh, Wen-Yuan; Chen, Ching-Yun; Liu, Chia-Ching; Lin, Feng-Huei

    2015-02-01

    Superparamagnetic nanoparticles (Fe3O4, SPIO) have been used as magnetic resonance imaging enhancers for years. However, bio-safety issues concerning nanoparticles remain largely unexplored. Of particular concern is the possible cellular impact of nanoparticles during SPIO uptake and subsequent oxidative stress. SPIO causes cell death by apoptosis via a little understood mitochondrial pathway. To more closely examine this process, three kinds of cells—3T3, RAW264.7, and MCF7—were treated with SPIO coated with polyethylene glycol (SPIO-PEG) and monitored by transmission electron microscopy (TEM), using cytotoxicity evaluation, mitochondrial activity, reactive oxygen species (ROS) generation, and Annexin V assay. TEM revealed that SPIO-PEG nanoparticles surrounded the cellular endosome membrane, creating a bulge in the endosome. Compared to 3T3 cells, greater numbers of SPIO-PEG nanoparticles infiltrated the mitochondria of RAW264.7 and MCF7 cells. SPIO-PEG residency is associated with boosted ROS, with elevated levels of mitochondrial activity, and advancement of cell apoptosis. Furthermore, correlation analysis showed that a polynomial model demonstrates a better fit than a linear model in MCF7, implying that cytotoxicity may have alternative impacts on cell death at different concentrations. Thus, we believe that MCF7 cell death results from the apoptosis pathway triggered by mitochondria, and we find lower cytotoxicity in 3T3. We propose that optimal levels of SPIO-PEG nanoparticles lead to increased levels of ROS and a resulting oxidative stress environment which will kill only cancer cells while sparing normal cells. This finding has great potential for use in cancer therapies in the future.

  16. Anti-inflammatory effects of cordycepin in lipopolysaccharide-stimulated RAW 264.7 macrophages through Toll-like receptor 4-mediated suppression of mitogen-activated protein kinases and NF-κB signaling pathways

    Directory of Open Access Journals (Sweden)

    Choi YH

    2014-10-01

    Full Text Available Yung Hyun Choi,1,2 Gi-Young Kim,3 Hye Hyeon Lee4 1Department of Biochemistry, Dongeui University College of Korean Medicine, Busan, 2Anti-Aging Research Center and Blue-Bio Industry RIC, Dongeui University, Busan, 3Laboratory of Immunobiology, Department of Marine Life Sciences, Jeju National University, Jeju, 4Daegu Gyeongbuk Institute of Science and Technology, Daegu, Republic of Korea Abstract: Cordycepin is the main functional component of the Cordyceps species, which has been widely used in traditional Oriental medicine. This compound possesses many pharmacological properties, such as an ability to enhance immune function, as well as antioxidant, antiaging, and anticancer effects. In the present study, we investigated the anti-inflammatory effects of cordycepin using a murine macrophage RAW 264.7 cell model. Our data demonstrated that cordycepin suppressed production of proinflammatory mediators such as nitric oxide (NO and prostaglandin E2 by inhibiting inducible NO synthase and cyclooxygenase-2 gene expression. Cordycepin also inhibited the release of proinflammatory cytokines, including tumor necrosis factor-alpha and interleukin-1-beta, through downregulation of respective mRNA expression. In addition, pretreatment with cordycepin significantly inhibited lipopolysaccharide (LPS-induced phosphorylation of mitogen-activating protein kinases and attenuated nuclear translocation of NF-κB by LPS, which was associated with abrogation of inhibitor kappa B-alpha degradation. Furthermore, cordycepin potently inhibited the binding of LPS to macrophages and LPS-induced Toll-like receptor 4 and myeloid differentiation factor 88 expression. Taken together, the results suggest that the inhibitory effects of cordycepin on LPS-stimulated inflammatory responses in RAW 264.7 macrophages are associated with suppression of mitogen-activating protein kinases and activation of NF-κB by inhibition of the Toll-like receptor 4 signaling pathway. Keywords

  17. Expression of Cyclooxygenase-2, Nitric Oxide Synthase 2 and Heme Oxygenase-1 mRNA Induced byBis-Eugenol in RAW264.7 Cells and their Antioxidant Activity Determined Using the Induction Period Method.

    Science.gov (United States)

    Murakami, Yukio; Kawata, Akifumi; Fujisawa, Seiichiro

    2017-01-01

    To clarify the mechanisms responsible for the anti-inflammatory/proinflammatory activities of eugenol-related compounds, we investigated the cytotoxicity and up-regulatory/down-refgulatory effects of the biphenols curcumin, bis-eugenol, magnolol and honokiol, and the monophenols eugenol and isoeugenol, on major regulators of cyclooxygenase-2 (Cox-2), nitric oxide synthase 2 (Nos2) and heme oxygenase-1 (HO-1) mRNA in RAW264.7 cells. mRNA expression was investigated using real-time reverse transcriptase-polymerase chain reaction (RT-PCR), and the theoretical parameters were calculated using the DFT/B3LYP/6-31* method. Also, the antioxidant activity of eugenol-related compounds in combination with 2-mercapto-1-methylimidazole (MMI, as a model for glutathione (GSH)) was investigated using the induction period method for polymerization of methyl methacrylate initiated by benzoyl peroxide (BPO). The cytotoxicity of eugenol-related compounds showed a linear relationship with their softness (σ) and electrophilicity (ω). At a concentration of 50 μM, biphenols except for bis-eugenol elicited the expression of mRNA for both Cox-2 and Nos2, but monophenols did not. In contrast, bis-eugenol elicited Cox-2 gene expression, but down-regulated Nos2 gene expression. bis-Eugenol alone induced the expression of HO-1 mRNA, and when combined with MMI it showed a potent antagonistic effect on BPO-induced antioxidant activity. The ability of methoxyphenols to inhibit LPS-stimulated Cox-2 gene expression declined in the order curcumin > isoeugenol > bis-eugenol > eugenol, and the rank of ability was related to their ω value. Most eugenol-related compounds had proinflammatory activity at high concentrations. However, they had also anti-inflammatory activity at lower concentrations. Eugenol-related compounds may exert antioxidant and anti-inflammatory activity in LPS-stimulated RAW264.7 cells possibly by inhibiting the activation of nuclear factor-kappa B (Nf-ĸB), whereas bis

  18. Determination of normal expression patterns of CD86, CD210a, CD261, CD262, CD264, CD358, and CD361 in peripheral blood and bone marrow cells by flow cytometry.

    Science.gov (United States)

    Rudolf-Oliveira, Renata Cristina Messores; Auat, Mariangeles; Cardoso, Chandra Chiappin; Santos-Pirath, Iris Mattos; Lange, Barbara Gil; Pires-Silva, Jéssica; Moraes, Ana Carolina Rabello de; Dametto, Gisele Cristina; Pirolli, Mayara Marin; Colombo, Maria Daniela Holthausen Périco; Santos-Silva, Maria Claudia

    2018-02-01

    In 2010, new monoclonal antibodies were submitted to the 9th International Workshop on Human Leukocyte Differentiation Antigens, and there are few studies demonstrating normal expression patterns of these markers. Thus, the objective of this study was to determine the normal patterns of cell expression of CD86, CD210a, CD261, CD262, CD264, CD358, and CD361 in peripheral blood (PB) and bone marrow (BM) samples by flow cytometry. In the present study, CD86 was expressed only in monocytes and B lymphocytes in PB and in monocytes and plasma cells in BM. Regarding CD210a expression, in PB samples, monocytes and NK cells showed weak expression, while neutrophils, B and T lymphocytes, and basophils showed weak and partial expression. In BM samples, expression of CD210a was observed in eosinophils, monocytes, and B and T/NK lymphocytes. Weak expression of CD210a was also observed in neutrophilic cells and plasma cells. All B cell maturation stages had weak expression of CD210a except for immature B cells, which did not express this marker. In the present study, no cell type in PB samples showed positivity for CD261 and, in BM samples, there was very weak expression in neutrophilic series, monocytes, and B lymphocytes. Conversely, plasma cells showed positivity for CD261 with a homogeneous expression. For CD262, there was weak expression in monocytes, neutrophils, and B lymphocytes in PB samples and weak expression in monocytes, B lymphocytes, and plasma cells in BM samples. The evaluation of CD264 showed very weak expression in B cells in PB samples and no expression in BM cells. Very weak expression of CD358 was observed in neutrophils, monocytes, and B lymphocytes in PB and BM samples. In addition, in BM samples, plasma cells and T lymphocytes showed weak expression of CD358. In relation to the maturation stages of B cells, there was weak expression in pro-B cel, pre-B cell, and mature B cell. In the present study, it was possible to observe expression of CD361 in all

  19. Inhibition of LDL oxidation and oxidized LDL-induced foam cell formation in RAW 264.7 cells show anti-atherogenic properties of a foliar methanol extract of Scoparia dulcis.

    Science.gov (United States)

    Nambiar, Sinjitha S; Shetty, Nandini Prasad; Bhatt, Praveena; Neelwarne, Bhagyalakshmi

    2014-04-01

    Oxidation of low density lipoproteins and their further uptake by macrophages is known to result in the formation of foam cells, which are critical in the initiation of atherosclerosis through activation of inflammatory signalling cascades. Thus, powerful dietary antioxidants are receiving attention for the reversal of such pathological states. Extracts of Scoparia dulcis have been used as tea and health drinks with various health promoting effects. In the present study, we examined the reactive oxygen scavenging potential as well as anti-inflammatory and anti-atherogenic efficacies, using leaf extracts obtained after successive extraction with various solvents. A methanol extract showed potent antioxidant activity with an IC50 value of 570 μg/ml, caused hydrogen peroxide scavenging (28.9 µg/ml) and anti-inflammatory effects by improving human erythrocyte membrane stabilisation (about 86%). The methanol extract also efficiently inhibited lipid peroxidation and oxidation of low density lipoproteins, thus preventing foam cell formation in cultured RAW 264.7 cells. Furthermore, phytochemical screening of the extracts showed high accumulation of flavonoids. The foliar methanol extract of Scoparia dulcis has a strong anti-atherogenic potential and this property could be attributed maybe due to presence of flavonoids since HPLC analysis showed high concentrations of myricetin and rutin in the methanol extract.

  20. TOP 1 and 2, polysaccharides from Taraxacum officinale, inhibit NFκB-mediated inflammation and accelerate Nrf2-induced antioxidative potential through the modulation of PI3K-Akt signaling pathway in RAW 264.7 cells.

    Science.gov (United States)

    Park, Chung Mu; Cho, Chung Won; Song, Young Sun

    2014-04-01

    Anti-inflammatory and anti-oxidative activities of polysaccharides from Taraxacum officinale (TOP 1 and 2) were analyzed in RAW 264.7 cells. First, lipopolysaccharide (LPS) was applied to identify anti-inflammatory activity of TOPs, which reduced expression of inducible nitric oxide synthase (iNOS) and tumor necrosis factor (TNF)-α. TOPs treatment inhibited phosphorylation of inflammatory transcription factor, nuclear factor (NF)κB, and its upstream signaling molecule, PI3K/Akt. Second, cytoprotective potential of TOPs against oxidative stress was investigated via heme oxygenase (HO)-1 induction. HO-1, one of phase II enzymes shows antioxidative activity, was potently induced by TOPs treatment, which was in accordance with the nuclear translocation of nuclear factor-erythroid 2 p45-related factor 2 (Nrf2). In addition, TOPs treatment phosphorylated PI3K/Akt with slight activation of c-Jun NH2-terminal kinase (JNK). TOPs-mediated HO-1 induction protected macrophage cells from oxidative stress-induced cell death, which was confirmed by SnPP and CoPP (HO-1 inhibitor and inducer, respectively). Consequently, TOPs potently inhibited NFκB-mediated inflammation and accelerated Nrf2-mediated antioxidative potential through the modulation of PI3K/Akt pathway, which would contribute to their promising strategy for novel anti-inflammatory and anti-oxidative agents. Copyright © 2014. Published by Elsevier Ltd.

  1. MicroRNA-144-3p inhibits autophagy activation and enhances Bacillus Calmette-Guérin infection by targeting ATG4a in RAW264.7 macrophage cells.

    Science.gov (United States)

    Guo, Le; Zhou, Linlin; Gao, Qian; Zhang, Aijun; Wei, Jun; Hong, Dantong; Chu, Yuankui; Duan, Xiangguo; Zhang, Ying; Xu, Guangxian

    2017-01-01

    MicroRNAs (miRNAs) are small noncoding nucleotides that play major roles in the response of host immune cells. Autophagy plays a key role in activating the antimicrobial host defense against Mycobacterium tuberculosis (M. tuberculosis). Whether miRNAs specifically influence the activation of macrophage autophagy during M. tuberculosis infection is largely unknown. In the present study, we demonstrate that Mycobacterium bovis Bacillus Calmette-Guérin (BCG) infection of macrophages leads to increased expression of miR-144-3p, which targets autophagy-related gene 4a (ATG4a), to inhibit autophagy activation and antimicrobial responses to BCG. Overexpression of miR-144-3p significantly decreased both mRNA and protein levels of ATG4a, inhibited the formation of autophagosomes in RAW264.7 cells and increased intracellular survival of BCG. However, transfection with miR-144-3p inhibitor led to an increase in ATG4a levels, accelerated the autophagic response in macrophages, and decreased BCG survival in macrophages. The experimental results of this study reveal a novel role of miR-144-3p in inhibiting autophagy activation by targeting ATG4a and enhancing BCG infection, and provide potential targets for developing improved treatment.

  2. Polysaccharide of Danggui Buxue Tang, an Ancient Chinese Herbal Decoction, Induces Expression of Pro-inflammatory Cytokines Possibly Via Activation of NFκB Signaling in Cultured RAW 264.7 Cells.

    Science.gov (United States)

    Gong, Amy Gw; Zhang, Laura Ml; Lam, Candy Tw; Xu, Miranda L; Wang, Huai Y; Lin, H Q; Dong, Tina Tx; Tsim, Karl Wk

    2017-02-01

    Danggui Buxue Tang (DBT) is an ancient Chinese herbal decoction containing two herbs, Astragali Radix (AR) and Angelicae Sinensis Radix (ASR): this herbal decoction serves as dietary supplement for women during menopause. DBT has been known to modulate immune responses, and its polysaccharide is proposed to be one of the active components. However, the polysaccharide-induced signaling in immune activation is not revealed. Here, we are identifying that the immune activation, triggered by DBT, could be mediated by polysaccharide. In cultured macrophages (RAW 264.7 cells), the application of polysaccharide-enriched extract of DBT significantly increased the expressions of mRNA and protein levels of interleukin-1β, interleukin-6 and tumor necrosis factor. The induction was much stronger than the polysaccharide extract generated singly from AR, or from ASR, or from their simple mixture. The induced cytokine release in cultured macrophage was revealed to be triggered by activation of nuclear factor-kappa B (NF-κB) signaling, including (i) degradation of IkBα; (ii) translocation of NF-κB p65 from cytosol to nuclei; and (iii) activation of NF-κB transcriptional elements. These results verified the possible role of DBT polysaccharide in modulating immune responses. Copyright © 2016 John Wiley & Sons, Ltd. Copyright © 2016 John Wiley & Sons, Ltd.

  3. Anti-inflammatory properties of fermented soy milk with Lactococcus lactis subsp. lactis S-SU2 in murine macrophage RAW264.7 cells and DSS-induced IBD model mice.

    Science.gov (United States)

    Kawahara, Miho; Nemoto, Maki; Nakata, Toru; Kondo, Saya; Takahashi, Hajime; Kimura, Bon; Kuda, Takashi

    2015-06-01

    Six lactic acid bacteria strains (four Lactobacillus plantarum strains and one each of Lactococcus lactis subsp. lactis and Pediococcus pentosaceus) have been isolated and shown to possess anti-oxidant activity. In this study, we determined their acid, bile, salt resistance, and adhesion activity on human enterocyte-like HT-29-Luc and Caco-2 cells. An isolate Lc. lactis S-SU2 showed highest bile resistance and adhesion activity compared to type strains. S-SU2 could ferment both 10% skimmed milk and soy milk while the type strain could not ferment soy milk. Soy milk fermented with S-SU2 showed an increased nitric oxide (NO) secretion in the mouse macrophage RAW264.7 cells without bacterial lipopolysaccharide (LPS). Furthermore, the inhibitory effects of the fermented soy milk on Escherichia coli O111 LPS-induced NO secretion were higher than those of fresh soy milk. Inflammatory bowel disease (IBD) was induced in mice fed either 5% (w/v) dextran sodium sulfate (DSS) in drinking water or 50% soy milk in drinking water. Shortening of colon length, breaking of epithelial cells, lowering liver and thymus weights, and enlargement of spleen are some of the characteristics observed in the IBD, which were prevented by the use of soy milk fermented with Lc. lactis S-SU2. Copyright © 2015 Elsevier B.V. All rights reserved.

  4. A molecular pharmacology study into the anti-inflammatory actions of Euphorbia hirta L. on the LPS-induced RAW 264.7 cells through selective iNOS protein inhibition.

    Science.gov (United States)

    Shih, Mei-Fen; Cheng, Yih-Dih; Shen, Chia-Rui; Cherng, Jong-Yuh

    2010-07-01

    Euphorbia hirta L. has been widely used in India and Chinese society. The molecular pharmacology basis of its anti-inflammatory effect is revealed in this work. The ethanol extract of Euphorbia hirta L. (Eh) and its active component were studied in lipopolysaccharide (LPS)-activated macrophage cells (RAW 264.7) as an established inflammation model. After activation, nitric oxide (NO) production and expression of iNOS protein and iNOS mRNA were measured by using a colorimetric assay (Griess reagent), western blotting, and reverse transcription polymerase chain reaction (RT-PCR), respectively. The alteration in the content of PGE(2), TNFalpha, and IL-6 was concurrently monitored by ELISA. In results, we found that in the concentration range without showing cytotoxicity, Eh produced a remarkable anti-inflammatory effect via its active component of beta-amyrin and showed a dose-related inhibition of LPS-induced NO production. This phenomenon is in accordance with a substantial inhibition of iNOS protein. However, the expression of iNOS gene was unaffected by Eh treatments. Compared with indomethacin, Eh has much more potency and a specific action of NO inhibition but Eh works less specifically on PGE(2), IL-6, and TNF-alpha inhibition. The extract of Euphorbia hirta L. and its component beta-amyrin are able to block most of the iNOS protein functions and NO induction, and could therefore be new selective NO inhibitors with great potential in treating arthritis inflammation.

  5. Phosphoproteome and transcription factor activity profiling identify actions of the anti-inflammatory agent UTL-5g in LPS stimulated RAW 264.7 cells including disrupting actin remodeling and STAT-3 activation.

    Science.gov (United States)

    Carruthers, Nicholas J; Stemmer, Paul M; Chen, Ben; Valeriote, Frederick; Gao, Xiaohua; Guatam, Subhash C; Shaw, Jiajiu

    2017-09-15

    UTL-5g is a novel small-molecule TNF-alpha modulator. It reduces cisplatin-induced side effects by protecting kidney, liver, and platelets, thereby increasing tolerance for cisplatin. UTL-5g also reduces radiation-induced acute liver toxicity. The mechanism of action for UTL-5g is not clear at the present time. A phosphoproteomic analysis to a depth of 4943 phosphopeptides and a luminescence-based transcription factor activity assay were used to provide complementary analyses of signaling events that were disrupted by UTL-5g in RAW 264.7 cells. Transcriptional activity downstream of the interferon gamma, IL-6, type 1 Interferon, TGF-β, PKC/Ca 2+ and the glucocorticoid receptor pathways were disrupted by UTL-5g. Phosphoproteomic analysis indicated that hyperphosphorylation of proteins involved in actin remodeling was suppressed by UTL-5g (gene set analysis, FDR 5g. This global characterization of UTL-5g activity in a macrophage cell line discovered that it disrupts selected aspects of LPS signaling including Stat3 activation and actin remodeling providing new insight on how UTL-5g acts to reduce cisplatin-induced side effects. Copyright © 2017 Elsevier B.V. All rights reserved.

  6. Anti-inflammatory action of 2-carbomethoxy-2,3-epoxy-3-prenyl-1,4-naphthoquinone (CMEP-NQ) suppresses both the MyD88-dependent and TRIF-dependent pathways of TLR4 signaling in LPS-stimulated RAW264.7 cells.

    Science.gov (United States)

    Ju Woo, Hyun; Jun, Do Youn; Lee, Ji Young; Park, Hae Sun; Woo, Mi Hee; Park, Sook Jahr; Kim, Sang Chan; Yang, Chae Ha; Kim, Young Ho

    2017-06-09

    The roots of Rubia cordifolia L. have been widely used as a traditional herbal medicine in Northeast Asia for treating inflammatory diseases. To elucidate the anti-inflammatory mechanism of 2-carbomethoxy-2,3-epoxy-3- prenyl-1,4-naphthoquinone (CMEP-NQ), purified from the roots of R. cordifolia L. as the major anti-inflammatory component, in LPS-treated RAW264.7 murine macrophage cells. Anti-inflammatory activity of CMEP-NQ was investigated in LPS-treated RAW264.7 cells by measuring the levels of NO, PGE 2 , and cytokines (IL1β, IL-6, TNF-α) in the culture supernatants and the TLR4-mediated intracellular events including association of MyD88 with IRAK1, activation of IRAK1, TAK1, MAPKs, NF-κB/AP-1, and IRF3, and generation of ROS. Pretreatment of RAW264.7 cells with CMEP-NQ reduced LPS-induced production of NO and PGE 2 by suppressing iNOS and COX-2 gene expression. CMEP-NQ also reduced the secretion of IL-1β, IL-6, and TNF-α by down-regulating mRNA levels. Under these conditions, TLR4-mediated MyD88-dependent events were inhibited by CMEP-NQ, including the association of MyD88 with IRAK1, phosphorylation of IRAK1, TAK1, and MAPKs (ERK, JNK and p38 MAPK), and activation of NF-κB and AP-1. As TRIF-dependent events of TLR4 signaling, phosphorylation of IRF3 and induction of iNOS protein expression were also inhibited by CMEP-NQ. However, the binding of FITC-conjugated LPS to cell surface TLR4 was not affected by CMEP-NQ. Following LPS stimulation, intracellular ROS production was first detected by DCFH-DA staining at 1h; then it continuously increased until 16h. Although CMEP-NQ failed to exhibit DPPH radical- or ABTS radical-scavenging activity in vitro, LPS-induced ROS production in RAW264.7 cells was more efficiently blocked by CMEP-NQ than by NAC. These results demonstrate that the suppressive effect of CMEP-NQ on LPS-induced inflammatory responses in RAW264.7 cells was mainly exerted via its inhibition of TLR4-mediated proximal events, such as MyD88

  7. A comparison of cytotoxicity and oxidative stress from welding fumes generated with a new nickel-, copper-based consumable versus mild and stainless steel-based welding in RAW 264.7 mouse macrophages.

    Science.gov (United States)

    Badding, Melissa A; Fix, Natalie R; Antonini, James M; Leonard, Stephen S

    2014-01-01

    Welding processes that generate fumes containing toxic metals, such as hexavalent chromium (Cr(VI)), manganese (Mn), and nickel (Ni), have been implicated in lung injury, inflammation, and lung tumor promotion in animal models. While federal regulations have reduced permissible worker exposure limits to Cr(VI), this is not always practical considering that welders may work in confined spaces and exhaust ventilation may be ineffective. Thus, there has been a recent initiative to minimize the potentially hazardous components in welding materials by developing new consumables containing much less Cr(VI) and Mn. A new nickel (Ni) and copper (Cu)-based material (Ni-Cu WF) is being suggested as a safer alternative to stainless steel consumables; however, its adverse cellular effects have not been studied. This study compared the cytotoxic effects of the newly developed Ni-Cu WF with two well-characterized welding fumes, collected from gas metal arc welding using mild steel (GMA-MS) or stainless steel (GMA-SS) electrodes. RAW 264.7 mouse macrophages were exposed to the three welding fumes at two doses (50 µg/ml and 250 µg/ml) for up to 24 hours. Cell viability, reactive oxygen species (ROS) production, phagocytic function, and cytokine production were examined. The GMA-MS and GMA-SS samples were found to be more reactive in terms of ROS production compared to the Ni-Cu WF. However, the fumes from this new material were more cytotoxic, inducing cell death and mitochondrial dysfunction at a lower dose. Additionally, pre-treatment with Ni-Cu WF particles impaired the ability of cells to phagocytize E. coli, suggesting macrophage dysfunction. Thus, the toxic cellular responses to welding fumes are largely due to the metal composition. The results also suggest that reducing Cr(VI) and Mn in the generated fume by increasing the concentration of other metals (e.g., Ni, Cu) may not necessarily improve welder safety.

  8. Effect of apigenin, kaempferol and resveratrol on the gene expression and protein secretion of tumor necrosis factor alpha (TNF-α) and interleukin-10 (IL-10) in RAW-264.7 macrophages.

    Science.gov (United States)

    Palacz-Wrobel, Marta; Borkowska, Paulina; Paul-Samojedny, Monika; Kowalczyk, Malgorzata; Fila-Danilow, Anna; Suchanek-Raif, Renata; Kowalski, Jan

    2017-09-01

    Polyphenols such as apigenin, kaempferol or resveratrol are typically found in plants, including fruits, vegetables, herbs and spices, which have a wide range of biological functions such as antioxidative, anti-inflammatory, vasodilative, anticoagulative and proapoptotic. Discovering such multifunctional compounds in widely consumed plant-based products - ones that both inhibit the release of TNF-α from tissue macrophages and at the same time enhance the secretion of IL-10 - would be an important signpost in the quest for effective pharmacological treatment of numerous diseases that have an inflammatory etiology. The aim of the study is to investigate the impact of biologically active polyphenols such as apigenin, resveratrol and kaempferol on gene expression and protein secretion of IL-10 and TNF-α in line RAW-264.7. Cells were cultured under standard conditions. IL-10 and TNF-α genes expression were examined using QRT-PCR and to assess cytokines concentration ELISA have been used. Apigenin, kaempferol and resveratrol at a dose 30μM significantly decrease the TNF-α expression and secretion. Apigenin decrease the IL-10 expression and secretion. Furthermore, increase in IL-10 secretion after administration of kaempferol and resveratrol were observed. In the process of administration of tested compounds before LPS, which activate macrophages, decrease of TNF-α secretion after apigenin and kaempferol and increase of IL-10 secretion after resveratrol were observed. The results of present work indicate that 1) apigenin, resveratrol and kaempferol may reduce the intensity of inflammatory processes by inhibiting the secretion of proinflammatory cytokine TNF-α, and resveratrol and kaempferol additionally by increasing the secretion of anti-inflammatory cytokine IL-10 2) the studies indicate the potentially beneficial - anti-inflammatory - impact of diet rich in products including apigenin, resveratrol and kaempferol. Copyright © 2017 Elsevier Masson SAS. All rights

  9. Mitochondrion-Targeted Peptide SS-31 Inhibited Oxidized Low-Density Lipoproteins-Induced Foam Cell Formation through both ROS Scavenging and Inhibition of Cholesterol Influx in RAW264.7 Cells

    Directory of Open Access Journals (Sweden)

    Shuangying Hao

    2015-12-01

    Full Text Available Foam cell formation as a result of imbalance of modified cholesterol influx and efflux by macrophages is a key to the occurrence and development of atherosclerosis. Oxidative stress is thought to be involved in the pathogenesis of atherosclerosis. SS-31 is a member of the Szeto-Schiller (SS peptides shown to specifically target the inner mitochondrial membrane to scavenge reactive oxygen species. In this study, we investigated whether SS-31 may provide protective effect on macrophage from foam cell formation in RAW264.7 cells. The results showed that SS-31 inhibited oxidized low-density lipoproteins (ox-LDL-induced foam cell formation and cholesterol accumulation, demonstrated by intracellular oil red O staining and measurement of cholesterol content. The mechanism was revealed that SS-31 did not only significantly attenuated ox-LDL-induced generation of reactive oxygen species (ROS and increased the activities of superoxide dismutases, but also dose-dependently inhibited the expression of CD36 and LOX-1, two scavenger receptors of ox-LDL, while the expression of ATP-binding cassette A1 and G1, playing a pivotal role in cholesterol efflux, was not affected. As a result, SS-31 decreased pro-inflammatory cytokines such as interleukin 6 and tumor necrosis factor alpha, suggesting the prevention of inflammatory responses. In conclusion, our results demonstrate that SS-31 provides a beneficial effect on macrophages from foam cell formation, likely, through both ROS scavenging and inhibition of cholesterol influx. Therefore, SS-31 may potentially be of therapeutic relevance in prevention of human atherogenesis.

  10. Andrographolide acts as an anti-inflammatory agent in LPS-stimulated RAW264.7 macrophages by inhibiting STAT3-mediated suppression of the NF-κB pathway.

    Science.gov (United States)

    Lee, Ko-Chen; Chang, Hen-Hong; Chung, Ying-Hui; Lee, Tzung-Yan

    2011-06-01

    Inflammation is involved in numerous diseases, such as chronic inflammatory disease and cancer. Many plant products exhibit useful biological activities, including antifungal, antibacterial, and anti-inflammatory actions. However, our understanding of the anti-inflammatory effects of andrographolide is limited. We use lipopolysaccharide (LPS)-stimulated macrophages as a model of inflammation to investigate the anti-inflammatory effects of andrographolide, which contains polyphenolic structures. We found that andrographolide exhibited a potent anti-inflammatory effect. In this study, we investigated the inhibitory effects of andrographolide on the induction of nitric oxide synthase (NOS) and cyclooxygenase-2 (COX-2) as well as their respective downstream products, NO and PGE2, in RAW264.7 cells treated with LPS. Treatment with andrographolide also reduced nuclear factor-κB (NF-κB) and activation protein-1 (AP-1) DNA-binding activity. Western blot analysis showed that andrographolide significantly inhibited the phosphorylation of signal transducer and activator of transcription-3 (STAT3) and the protein expression of CCAAT/enhancer-binding protein δ (C/EBPδ). We also found that andrographolide suppressed LPS-induced suppressor of cytokine signalling 1 and 3 (SOCS1 and 3) mRNA expression, which, in turn, inhibited apoptosis signalling and mitochondria membrane potential activation. Our results demonstrate that andrographolide downregulates inflammatory iNOS and COX-2 gene expression by inhibiting the activation of NF-κB and STAT3 by interfering with the expression of SOCS1 and SOCS3 signalling. Therefore, andrographolide exerts a potent anti-inflammatory effect and could potentially be developed as a useful agent for the chemoprevention of cancer or inflammatory diseases. Copyright © 2011 Elsevier Ireland Ltd. All rights reserved.

  11. The heme oxygenase-1 inducer THI-56 negatively regulates iNOS expression and HMGB1 release in LPS-activated RAW 264.7 cells and CLP-induced septic mice.

    Science.gov (United States)

    Park, Eun Jung; Jang, Hwa Jin; Tsoyi, Konstantin; Kim, Young Min; Park, Sang Won; Kim, Hye Jung; Lee, Jae Heun; Chang, Ki Churl

    2013-01-01

    The nuclear DNA binding protein high mobility group box 1 (HMGB1) has recently been suggested to act as a late mediator of septic shock. The effect of ((S)-6,7-dihydroxy-1-(4-hydroxynaphthylmethyl)-1,2,3,4-tetrahydroisoquinoline alkaloid, also known as THI-56, in an experimental model of sepsis was investigated. THI-56 exhibited potent anti-inflammatory properties in response to LPS in RAW 264.7 cells. In particular, THI-56 significantly inhibited the expression of inducible nitric oxide synthase (iNOS) and the release of HMGB1 in activated macrophages. THI-56 activated NE-F2-regulated factor 2 (Nrf-2)/heme oxygenase 1 (HO-1). The specific knockdown of the HO-1 gene by HO-1 siRNA significantly reversed the inhibitory effects of THI-56 on iNOS expression and HMGB1 release in LPS-stimulated macrophages. Importantly, THI-56 administration protected animals from death induced by either a lethal dose of LPS or cecal ligation and puncture (CLP). Furthermore, the ALT, AST, BUN, creatinine, and HMGB1 levels in the blood were significantly increased in CLP-induced septic mice, and the administration of THI-56 reduced these levels in a concentration-dependent and zinc protoporphyrin IX (ZnPPIX)-sensitive manner. In addition, the administration of THI-56 significantly ameliorated not only lung damage but also macrophage infiltration in the livers of CLP-induced septic mice, and these effects were also abrogated in the presence of ZnPPIX. Thus, we conclude that THI-56 significantly attenuates the proinflammatory response induced by LPS and reduces organ damage in a CLP-induced sepsis model through the upregulation of Nrf-2/HO-1.

  12. A comparison of cytotoxicity and oxidative stress from welding fumes generated with a new nickel-, copper-based consumable versus mild and stainless steel-based welding in RAW 264.7 mouse macrophages.

    Directory of Open Access Journals (Sweden)

    Melissa A Badding

    Full Text Available Welding processes that generate fumes containing toxic metals, such as hexavalent chromium (Cr(VI, manganese (Mn, and nickel (Ni, have been implicated in lung injury, inflammation, and lung tumor promotion in animal models. While federal regulations have reduced permissible worker exposure limits to Cr(VI, this is not always practical considering that welders may work in confined spaces and exhaust ventilation may be ineffective. Thus, there has been a recent initiative to minimize the potentially hazardous components in welding materials by developing new consumables containing much less Cr(VI and Mn. A new nickel (Ni and copper (Cu-based material (Ni-Cu WF is being suggested as a safer alternative to stainless steel consumables; however, its adverse cellular effects have not been studied. This study compared the cytotoxic effects of the newly developed Ni-Cu WF with two well-characterized welding fumes, collected from gas metal arc welding using mild steel (GMA-MS or stainless steel (GMA-SS electrodes. RAW 264.7 mouse macrophages were exposed to the three welding fumes at two doses (50 µg/ml and 250 µg/ml for up to 24 hours. Cell viability, reactive oxygen species (ROS production, phagocytic function, and cytokine production were examined. The GMA-MS and GMA-SS samples were found to be more reactive in terms of ROS production compared to the Ni-Cu WF. However, the fumes from this new material were more cytotoxic, inducing cell death and mitochondrial dysfunction at a lower dose. Additionally, pre-treatment with Ni-Cu WF particles impaired the ability of cells to phagocytize E. coli, suggesting macrophage dysfunction. Thus, the toxic cellular responses to welding fumes are largely due to the metal composition. The results also suggest that reducing Cr(VI and Mn in the generated fume by increasing the concentration of other metals (e.g., Ni, Cu may not necessarily improve welder safety.

  13. Synthesis of ceramic powders of La{sub 9,56} (SiO{sub 4}){sub 6}O{sub 2,34} and La{sub 9,8}Si{sub 5,7}Mg{sub O,3}O{sub 26,}4 by modified sol-gel process; Sintese de pos ceramicos de La{sub 9,56} (SiO{sub 4}){sub 6}O{sub 2,34} e La{sub 9,8}Si{sub 5,7}Mg{sub O,3}O{sub 26,}4 por processo sol-gel modificado

    Energy Technology Data Exchange (ETDEWEB)

    Lira, Sabrina Lopes; Paiva, Mayara Rafaela Soares; Misso, Agatha Matos; Elias, Daniel Ricco; Yamagata, Chieko, E-mail: yamagata@ipen.br [Instituto de Pesquisas Energeticas e Nucleares (CCTM/IPEN/CNEN-SP), Sao Paulo, SP (Brazil). Centro de Ciencia e Tecnologia de Materiais

    2012-07-01

    Lanthanum silicate oxyapatite materials are promising for application as electrolyte in solid oxide fuel cells because of high ionic conductivity at temperatures between 600 deg C and 800 deg C. In this work, oxyapatites with the composition La{sub 9,56}(SiO{sub 4}){sub 6}O{sub 2,34}, and La{sub 9,8}Si{sub 5,7}Mg{sub 0,3}O{sub 26,4} were synthesized by using the sol-gel method, followed by precipitation. Initially, the gel of silica was synthesized from sodium silicate solution, by acid catalysis using lanthanum and magnesium chloride solution. Then, the La and Mg hydroxides were precipitated with NaOH in the gel. The powders were characterized by X-ray diffraction (XRD), scanning electron microscopy (SEM) and measurements of specific surface area. The crystalline oxyapatite phase of La{sub 9,56}(SiO{sub 4}){sub 6}O{sub 2,34}, and was La{sub 9,8}Si{sub 5,7}Mg{sub 0,3}O{sub 26,4} obtained by calcination at 900 deg C for 2 and 1h respectively (author)

  14. 12 CFR 264b.2 - Definitions.

    Science.gov (United States)

    2010-01-01

    ... the United States at the time of acceptance of $285 or less as of January 1, 2002, and at 3-year... consultation with the Secretary of State, to reflect changes in the consumer price index for the immediately...

  15. 40 CFR 264.72 - Manifest discrepancies.

    Science.gov (United States)

    2010-07-01

    ... is impossible to locate an alternative facility that can receive the waste, the facility may return the rejected waste or residue to the generator. The facility must send the waste to the alternative... under this section, it must ensure that either the delivering transporter retains custody of the waste...

  16. 40 CFR 264.1064 - Recordkeeping requirements.

    Science.gov (United States)

    2010-07-01

    ... (CONTINUED) STANDARDS FOR OWNERS AND OPERATORS OF HAZARDOUS WASTE TREATMENT, STORAGE, AND DISPOSAL FACILITIES... device other than a thermal vapor incinerator, catalytic vapor incinerator, flare, boiler, process heater, condenser, or carbon adsorption system, the Regional Administrator will specify the appropriate...

  17. 32 CFR 264.3 - Background.

    Science.gov (United States)

    2010-07-01

    ... Information for Defense Purposes with Australia, Belgium, Denmark, France, the Federal Republic of Germany, Greece, Italy, Japan, The Netherlands, Norway, Portugal, Spain, Turkey, and the United Kingdom. The... Organization and European Regional Organizations (USRO), Paris, France, is the United States representative to...

  18. 40 CFR 264.18 - Location standards.

    Science.gov (United States)

    2010-07-01

    ... consider them in planning the location of a facility to help prevent subsequent project delays.] (c) Salt dome formations, salt bed formations, underground mines and caves. The placement of any noncontainerized or bulk liquid hazardous waste in any salt dome formation, salt bed formation, underground mine or...

  19. 40 CFR 264.1031 - Definitions.

    Science.gov (United States)

    2010-07-01

    ... each valve, pump, compressor, pressure relief device, sampling connection system, open-ended valve or line, or flange or other connector, and any control devices or systems required by this subpart. Flame... system, or through a tank (e.g., distillate receiver, condenser, bottoms receiver, surge control tank...

  20. 40 CFR 264.570 - Applicability.

    Science.gov (United States)

    2010-07-01

    ... existing drip pads to convey treated wood drippage, precipitation, and/or surface water run-off to an... provides protection from precipitation so that neither run-off nor run-on is generated is not subject to... owner or operator maintains and complies with a written contingency plan that describes how the owner or...

  1. 40 CFR 264.1084 - Standards: Tanks.

    Science.gov (United States)

    2010-07-01

    ... wells, and stub drains. (C) Each penetration of the internal floating roof for the purpose of sampling... the liquid in the tank and the other end extends a vertical distance of at least 61 centimeters above..., roof drains, and leg sleeves, each opening in the roof shall be equipped with a gasketed cover, seal...

  2. 40 CFR 264.554 - Staging piles.

    Science.gov (United States)

    2010-07-01

    ... by a qualified Professional Engineer for technical data, such as design drawings and specifications... facility that may influence the migration of any potential releases; and (vi) Potential for human and...

  3. 40 CFR 264.90 - Applicability.

    Science.gov (United States)

    2010-07-01

    ...), that the treatment zone of a land treatment unit that qualifies as a regulated unit does not contain... care period; or (4) The Regional Administrator finds that there is no potential for migration of liquid... must be certified by a qualified geologist or geotechnical engineer. In order to provide an adequate...

  4. 21 CFR 26.4 - Product coverage.

    Science.gov (United States)

    2010-04-01

    ...), investigational medicinal products/new drugs, human radiopharmaceuticals, and medicinal gases are also excluded... this subpart will apply to medicinal products for human or animal use, intermediates and starting...

  5. 40 CFR 264.1100 - Applicability.

    Science.gov (United States)

    2010-07-01

    ... handling equipment within the unit; (c) If the unit is used to manage liquids, has: (1) A primary barrier... to ensure containment and prevent the tracking of materials from the unit by personnel or equipment.... The owner or operator is not subject to the definition of land disposal in RCRA section 3004(k...

  6. 40 CFR 264.1086 - Standards: Containers.

    Science.gov (United States)

    2010-07-01

    ... container that meets the applicable U.S. Department of Transportation (DOT) regulations on packaging... integral part of the container structural design (e.g., a “portable tank” or bulk cargo container equipped... Transportation (DOT) regulations on packaging hazardous materials for transportation as specified in paragraph (f...

  7. 40 CFR 264.1050 - Applicability.

    Science.gov (United States)

    2010-07-01

    ... Act permit issued pursuant to 40 CFR 52.2454. (h) Purged coatings and solvents from surface coating operations subject to the national emission standards for hazardous air pollutants (NESHAP) for the surface...) STANDARDS FOR OWNERS AND OPERATORS OF HAZARDOUS WASTE TREATMENT, STORAGE, AND DISPOSAL FACILITIES Air...

  8. Publications | Page 264 | IDRC - International Development ...

    International Development Research Centre (IDRC) Digital Library (Canada)

    IDRC works with developing-country researchers and institutions to build local capacity through funding, knowledge sharing, and training. Through books, articles, research publications, and studies, we aim to widen the impact of our investment and advance development research. We share the results of our funded ...

  9. 40 CFR 264.1080 - Applicability.

    Science.gov (United States)

    2010-07-01

    ... section applies only to the facility commonly referred to as the OSi Specialties Plant, located on State... unit process vents and the wastewater treatment system (using the EPA Water 8 model or other model...

  10. 40 CFR 264.94 - Concentration limits.

    Science.gov (United States)

    2010-07-01

    ... ground-water users; (v) The current and future uses of ground water in the area; (vi) The existing quality of ground water, including other sources of contamination and their cumulative impact on the ground-water quality; (vii) The potential for health risks caused by human exposure to waste constituents...

  11. 40 CFR 264.93 - Hazardous constituents.

    Science.gov (United States)

    2010-07-01

    ... rates of ground-water users; (v) The current and future uses of ground water in the area; (vi) The existing quality of ground water, including other sources of contamination and their cumulative impact on the ground-water quality; (vii) The potential for health risks caused by human exposure to waste...

  12. Kaempferol 7-O-β-D-glucoside isolated from the leaves of Cudrania tricuspidata inhibits LPS-induced expression of pro-inflammatory mediators through inactivation of NF-κB, AP-1, and JAK-STAT in RAW 264.7 macrophages.

    Science.gov (United States)

    Lee, Seung-Bin; Shin, Ji-Sun; Han, Hee-Soo; Lee, Hwi-Ho; Park, Jong Cheol; Lee, Kyung-Tae

    2018-03-25

    Kaempferol 7-O-β-D-glucoside (KPG), a natural flavonol isolated from Cudrania tricuspidata, has been reported to exert anti-cancer effects; however, its anti-inflammatory effects have not yet been reported. In this study, we demonstrate the suppressive effect of KPG on the production of nitric oxide (NO), prostaglandin E 2 (PGE 2 ), tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and interleukin-6 (IL-6) in lipopolysaccharide (LPS)-induced RAW 264.7 macrophages and mouse bone marrow-derived macrophages. KPG downregulated the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) at the protein level and iNOS, COX-2, TNF-α, IL-1β, and IL-6 at the mRNA level in LPS-treated RAW 264.7 macrophages. Moreover, we elucidated the underlying molecular mechanism, demonstrating that KPG attenuated LPS-induced nuclear factor-κB (NF-κB) activation by decreasing p65 nuclear translocation, inhibiting κBα (IκBα) phosphorylation/degradation and IκB kinaseα/β (IKKα/β) phosphorylation. KPG additionally reduced LPS-induced activator protein-1 (AP-1) activity by inhibiting c-Fos expression in the nucleus, though c-Jun was not affected. Furthermore, we revealed that KPG significantly abrogated the LPS-induced phosphorylation of signal transducer and activator of transcription (STAT) 1 (Ser 727, Tyr 701) and STAT3 (Tyr 705) through inhibiting the phosphorylation of Janus kinase (JAK) 1 and JAK2, its upstream activating proteins. Taken together, our data suggest that KPG induces anti-inflammatory activity by blocking NF-κB, AP-1, and JAK-STAT signaling pathways in LPS-treated RAW 264.7 macrophages, thus suppressing inflammatory mediators. Copyright © 2018 Elsevier B.V. All rights reserved.

  13. Synthesis of ceramic powders of La9,56 (SiO4)6O2,34 and La9,8Si5,7MgO,3O26,4 by modified sol-gel process

    International Nuclear Information System (INIS)

    Lira, Sabrina Lopes; Paiva, Mayara Rafaela Soares; Misso, Agatha Matos; Elias, Daniel Ricco; Yamagata, Chieko

    2012-01-01

    Lanthanum silicate oxyapatite materials are promising for application as electrolyte in solid oxide fuel cells because of high ionic conductivity at temperatures between 600 deg C and 800 deg C. In this work, oxyapatites with the composition La 9,56 (SiO 4 ) 6 O 2,34 , and La 9,8 Si 5,7 Mg 0,3 O 26,4 were synthesized by using the sol-gel method, followed by precipitation. Initially, the gel of silica was synthesized from sodium silicate solution, by acid catalysis using lanthanum and magnesium chloride solution. Then, the La and Mg hydroxides were precipitated with NaOH in the gel. The powders were characterized by X-ray diffraction (XRD), scanning electron microscopy (SEM) and measurements of specific surface area. The crystalline oxyapatite phase of La 9,56 (SiO 4 ) 6 O 2,34 , and was La 9,8 Si 5,7 Mg 0,3 O 26,4 obtained by calcination at 900 deg C for 2 and 1h respectively (author)

  14. Preparation and characterization of Bi26-2xMn2xMo10O69-d and Bi26.4Mn0.6Mo10-2yMe2yO69-d (Me = V, Fe Solid Solutions

    Directory of Open Access Journals (Sweden)

    Z. A. Mikhaylovskaya

    2017-09-01

    Full Text Available Single phase samples of bismuth molybdate, Bi26Mo10O69, doped with Mn on the bismuth sublattice and V, Fe on the molybdenum sublattice were found to crystallize in the triclinic Bi26Mo10O69 structure at low doping levels and in the monoclinic Bi26Mo10O69 structure - at higher dopant concentration. The assumption that all Mn ions have an oxidation state of +2 was confirmed by means of magnetic measurement results analysis using Curie-Weiss law. Conductivity was investigated using impedance spectroscopy. The conductivity of Bi26.4Mn0.6Mo9.6Fe0.4O69-d was 1.2*10-2 S*cm-1 at 973 K and 2.2*10-4 S*cm-1 at 623 K, and the conductivity of Bi26.4Mn0.6Mo9.2V0.8O69-d was 2.2*10-3 S*cm-1 at 973 K and 2.2*10-5 S*cm-1 at 623 K.

  15. Ethyl acetate extract from Asparagus cochinchinensis exerts anti-inflammatory effects in LPS-stimulated RAW264.7 macrophage cells by regulating COX-2/iNOS, inflammatory cytokine expression, MAP kinase pathways, the cell cycle and anti-oxidant activity

    Science.gov (United States)

    Lee, Hyun Ah; Koh, Eun Kyoung; Sung, Ji Eun; Kim, Ji Eun; Song, Sung Hwa; Kim, Dong Seob; Son, Hong Joo; Lee, Chung Yeoul; Lee, Hee Seob; Bae, Chang Joon; Hwang, Dae Youn

    2017-01-01

    Asparagus cochinchinesis (A. cochinchinesis) is a medicine traditionally used to treat fever, cough, kidney disease, breast cancer, inflammatory disease and brain disease in northeast Asian countries. Although numerous studies of the anti-inflammatory effects of A. cochinchinesis have been conducted, the underlying mechanisms of such effects in macrophages remain to be demonstrated. To investigate the mechanism of suppressive effects on the inflammatory response in macrophages, alterations of the nitric oxide (NO) level, the cell viability, inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) expression levels, inflammatory cytokine expression, the mitogen-activated protein kinase (MAPK) signaling pathway, cell cycle arrest and reactive oxygen species (ROS) levels were measured in lipopolysaccharide (LPS)-activated RAW264.7 cells following treatment with ethyl acetate extract from A. cochinchinesis root (EaEAC). RAW264.7 cells pretreated two different concentrations of EaEAC prior to LPS treatment exhibited no significant toxicity. The concentration of NO was significantly decreased in the EaEAC + LPS treated group compared with the vehicle + LPS treated group. A similar decrease in mRNA transcript level of COX-2, iNOS, pro-inflammatory cytokines [tumor necrosis factor-α and interleukin (IL)-1β] and anti-inflammatory cytokines (IL-6 and IL-10) was detected in the EaEAC + LPS treated group compared with the vehicle + LPS treated group, although the decrease rate varied. Enhancement of the phosphorylation of MAPK family members following LPS treatment was partially rescued in the EaEAC pretreated group, and the cell cycle was arrested at the G2/M phase. Furthermore, the EaEAC pretreated group exhibited a reduced level of ROS generation compared with the vehicle + LPS treated group. Taken together, these results suggest that EaEAC suppresses inflammatory responses through inhibition of NO production, COX-2 expression and ROS production, as well as

  16. Ethyl acetate extract from Asparagus cochinchinensis exerts anti‑inflammatory effects in LPS‑stimulated RAW264.7 macrophage cells by regulating COX‑2/iNOS, inflammatory cytokine expression, MAP kinase pathways, the cell cycle and anti-oxidant activity.

    Science.gov (United States)

    Lee, Hyun Ah; Koh, Eun Kyoung; Sung, Ji Eun; Kim, Ji Eun; Song, Sung Hwa; Kim, Dong Seob; Son, Hong Joo; Lee, Chung Yeoul; Lee, Hee Seob; Bae, Chang Joon; Hwang, Dae Youn

    2017-04-01

    Asparagus cochinchinesis (A. cochinchinesis) is a medicine traditionally used to treat fever, cough, kidney disease, breast cancer, inflammatory disease and brain disease in northeast Asian countries. Although numerous studies of the anti‑inflammatory effects of A. cochinchinesis have been conducted, the underlying mechanisms of such effects in macrophages remain to be demonstrated. To investigate the mechanism of suppressive effects on the inflammatory response in macrophages, alterations of the nitric oxide (NO) level, the cell viability, inducible nitric oxide synthase (iNOS) and cyclooxygenase‑2 (COX‑2) expression levels, inflammatory cytokine expression, the mitogen-activated protein kinase (MAPK) signaling pathway, cell cycle arrest and reactive oxygen species (ROS) levels were measured in lipopolysaccharide (LPS)-activated RAW264.7 cells following treatment with ethyl acetate extract from A. cochinchinesis root (EaEAC). RAW264.7 cells pretreated two different concentrations of EaEAC prior to LPS treatment exhibited no significant toxicity. The concentration of NO was significantly decreased in the EaEAC + LPS treated group compared with the vehicle + LPS treated group. A similar decrease in mRNA transcript level of COX‑2, iNOS, pro-inflammatory cytokines [tumor necrosis factor‑α and interleukin (IL)‑1β] and anti‑inflammatory cytokines (IL‑6 and IL‑10) was detected in the EaEAC + LPS treated group compared with the vehicle + LPS treated group, although the decrease rate varied. Enhancement of the phosphorylation of MAPK family members following LPS treatment was partially rescued in the EaEAC pretreated group, and the cell cycle was arrested at the G2/M phase. Furthermore, the EaEAC pretreated group exhibited a reduced level of ROS generation compared with the vehicle + LPS treated group. Taken together, these results suggest that EaEAC suppresses inflammatory responses through inhibition of NO production, COX‑2 expression

  17. Distribution of CCR5-Delta32, CCR5 promoter 59029 A/G, CCR2-64I and SDF1-3’A genetic polymorphisms in HIV-1 infected and uninfected patients in the West Region of Cameroon

    Science.gov (United States)

    2013-01-01

    Background Genetic variants of the genes encoding Human Immunodeficiency Virus-1 (HIV-1) co-receptors and their ligands, like CC-Chemokine Receptor 5 delta 32 mutation (CCR5-Delta32), CCR5 promoter A/G (Adenine/Guanine), CC-Chemokine Receptor 2 mutation 64 isoleucine (CCR2-64I) and the Stromal cell-derived Factor 3’A mutation (SDF1-3’A), are involved in the susceptibility to HIV-1 infection and progression. The prevalence of these mutations varies by Region. However, little is known about their distribution in the population of Dschang, located in the West Region of Cameroon. The prevalence of HIV in the West Region of Cameroon is lower than elsewhere in Cameroon. The objectives of this study were to determine the distribution of four AIDS Related Gene (ARG) variants in HIV-infected and non-infected population of Cameroon especially in the West Region and to estimate the contribution of these variants to the susceptibility or resistance to HIV infection. We also aimed to evaluate the effectiveness of genotyping using dried blood spot (DBS) samples. Methods A total of 179 participants were recruited from two hospitals in Dschang in the West Region of Cameroon. Their genotypes for CCR5-Delta32, CCR5 promoter 59029A/G, CCR2-64I and SDF1-3’A were analyzed using polymerase chain reaction (PCR) and restriction fragment length polymorphisms. Results A total of 179 participants were enrolled in the study. Among them, 32 (17.9%) were HIV positive and 147 (82.1%) were HIV negative. The allelic frequencies of these genes were: 0%, 49.72%, 17.6% and 100% respectively for CCR5-Delta32, CCR5 promoter 59029A/G, CCR2-64I and SDF1-3’A. No individual was found to carry the CCR5-Delta 32 mutation. All participants recruited were heterozygous for the SDF1-3’A allele. Conclusion Our data suggest that the CCR5-Delta32 cannot account for the protection as it was completely absent in our population. SDF1-3’A variants, may be in association with other polymorphisms, may account

  18. [Role of bone marrow tyrosine kinase on chromosome X in the production of pro-inflammatory cytokines from mouse mononuclear-macrophages RAW264.7 induced by endotoxin/lipopolysaccharide and its mechanism].

    Science.gov (United States)

    Fang, X; Hu, Y; Wang, Y; Liu, S; Wang, F; Chen, X L

    2017-04-20

    Objective: To investigate the role of bone marrow tyrosine kinase on chromosome X (BMX) in the production of tumor necrosis factor α (TNF-α) and interleukin-1β (IL-1β) from mouse mononuclear-macrophages induced by endotoxin/lipopolysaccharide (LPS) and its related mechanism. Methods: Mouse mononuclear-macrophages RAW264.7 were inoculated in 6-well plates and routinely cultured for the following experiments. (1) Cells were collected and divided into blank control group, LPS control group, and 75, 750, 7 500, 75 000 nmol/L BMX-IN-1 pretreatment groups according to the random number table, with 8 wells in each group. Cells in blank control group were routinely cultured for 25 h. Cells in LPS control group were routinely cultured for 24 h and stimulated by LPS in the final mass concentration (the same below) of 0.1 μg/mL for 1 h. Cells in the latter 4 groups were pretreated with BMX-IN-1 in the final molarity (the same below) of 75, 750, 7 500, 75 000 nmol/L for 24 h and stimulated by 0.1 μg/mL LPS for 1 h. The mRNA expression of TNF-α of cells in each group was determined by real-time fluorescent quantitative reverse transcription polymerase chain reaction (RT-PCR) to screen the optimum concentration of BMX-IN-1. (2) Cells were collected and divided into LPS control group and 2, 4, 8, 12, 18 h BMX-IN-1 pretreatment groups according to the random number table, with 8 wells in each group. Cells in LPS control group were stimulated by 0.1 μg/mL LPS for 1 h. Cells in the latter 5 groups were pretreated with optimum concentration of BMX-IN-1 for 2, 4, 8, 12, 18 h respectively and stimulated by 0.1 μg/mL LPS for 1 h. The mRNA expression of TNF-α of cells in each group was determined by real-time fluorescent quantitative RT-PCR to screen the optimum time for BMX-IN-1 pre-treatment. (3) Cells were collected and divided into blank control group, BMX-IN-1 control group, LPS control group, and BMX-IN-1+ LPS group according to the random number table, with 16 wells in

  19. BPU Simulator

    DEFF Research Database (Denmark)

    Rehr, Martin; Skovhede, Kenneth; Vinter, Brian

    2013-01-01

    in that process. Our goal is to support all execution platforms, and in this work we introduce the Bohrium Processing Unit, BPU, which will be the FPGA backend for Bohrium. The BPU is modeled as a PyCSP application, and the clear advantages of using CSP for simulating a new CPU is described. The current Py......CSP simulator is able to simulate 220 Monte Carlo simulations in less than 35 seconds in the smallest BPU simulation....

  20. 40 CFR 264.1034 - Test methods and procedures.

    Science.gov (United States)

    2010-07-01

    .... In the event that a sample is accidentally lost or conditions occur in which one of the three runs... pipe or other closed system that is used to transfer the waste after generation to the first affected...

  1. BKR 26(4) pp. 101-106 (Banjo)

    African Journals Online (AJOL)

    Femi J. Olorunniji

    2014-12-31

    Dec 31, 2014 ... convert starch or disaccharides to ethanol and proteins. (Abate et al., Gonzalez et al., 1998). Also a 2.5 fold increase in amylases by mixed cultures of Bacillus and Zymomonas mobilis has been reported.( Abate et al., 1999) There are available reports on amylase production using a combination of bacteria ...

  2. BKR 26(4) pp. 130-139 (Somade)

    African Journals Online (AJOL)

    Femi J. Olorunniji

    2014-12-31

    Dec 31, 2014 ... Biokemistri. An International Journal of the Nigerian Society for Experimental Biology ... U.S. Environmental Protection Agency (EPA) showed that cyfluthrin ..... Occupational and. Environmental Medicine 67:341–347. Cox, C. (1994) Cyfluthrin; insecticide fact sheet. Journal of. Pesticide Reform 14:28-34.

  3. BKR 26(4) pp. 124-129 (Achuba)

    African Journals Online (AJOL)

    Femi J. Olorunniji

    2014-12-31

    Dec 31, 2014 ... the animal house of Department of Anatomy, Delta State. University, Abraka. The experimental rats were housed in clean wooden cages and left to .... Lipid peroxidation, superoxide dismutase and catalse activities in rabbit fed petroleum contaminated diet. Pakistain Journal of Nutrition, 4(5): 330–335.

  4. 40 CFR 264.552 - Corrective Action Management Units (CAMU).

    Science.gov (United States)

    2010-07-01

    ... hazardous wastes, and all media (including ground water, surface water, soils, and sediments) and debris... difficulty of treatment and analysis of constituents with similar treatment properties. (5) Except as... in place after closure; (C) Potential for releases from the CAMU; (D) Physical and chemical...

  5. 40 CFR 264.1085 - Standards: Surface impoundments.

    Science.gov (United States)

    2010-07-01

    ... materials of construction and designing the cover and closure devices shall include: Organic vapor... closure device in the closed position, as applicable. (B) To remove accumulated sludge or other residues... construction and designing the cover and closure devices shall include: Organic vapor permeability; the effects...

  6. Efficient architecture for global elimination algorithm for H. 264 ...

    Indian Academy of Sciences (India)

    This paper presents a fast block matching motion esti mation algorithm and its architecture. The proposed architecture is based on Global Elimination (GE) Algorithm, which uses pixel averaging to reduce complexity of motion search while keeping performance close to that of full search. GE uses a preprocessing stage ...

  7. 264 Chemical Speciation of Chromium in Various Matrices in South ...

    African Journals Online (AJOL)

    Ignacy cukrowski

    production of chromium metal and ferroalloys, in the use of chromate solutions for electroplating and in the ... involving DTPA was previously optimised mainly for seawater samples16, 19, 28, 36 and water samples with ... 2.1. Apparatus. Adsorptive stripping voltammetric measurements were performed using a Metrohm.

  8. All projects related to | Page 264 | IDRC - International Development ...

    International Development Research Centre (IDRC) Digital Library (Canada)

    2012-05-01

    Women face many barriers to political participation in conflict zones across South Asia, including laws that perpetuate a culture of impunity and lack of justice for victims of violence and abuse by security forces. Start Date: May 1, 2012. End Date: August 19, 2015. Topic: VIOLENCE AGAINST WOMEN, POLITICAL ...

  9. 40 CFR 264.342 - Principal organic hazardous constituents (POHCs).

    Science.gov (United States)

    2010-07-01

    ...) Principal Organic Hazardous Constituents (POHCs) in the waste feed must be treated to the extent required by... organic constituents in the waste and on their concentration or mass in the waste feed, considering the... (CONTINUED) SOLID WASTES (CONTINUED) STANDARDS FOR OWNERS AND OPERATORS OF HAZARDOUS WASTE TREATMENT, STORAGE...

  10. 40 CFR 264.193 - Containment and detection of releases.

    Science.gov (United States)

    2010-07-01

    ..., the type of corrosion or erosion protection used, the rate of corrosion or erosion observed during the... wastes or accumulated liquid out of the system to the soil, ground water, or surface water at any time... facility, including the thickness of soils present between the tank system and ground water; and (iv) All...

  11. Efficient architecture for global elimination algorithm for H.264 ...

    Indian Academy of Sciences (India)

    a high compression ratio, a Block Matching Motion Esti- mation (BMME) algorithm is used to remove .... sum engine receives eight incoming pixels as shown on the left-hand side. Two such engines are needed to cal- ... corresponding block sum variable assignment to minimize multiplexing logic. The select signals for the ...

  12. 40 CFR 264.19 - Construction quality assurance program.

    Science.gov (United States)

    2010-07-01

    ...) Geomembranes (flexible membrane liners); (v) Leachate collection and removal systems and leak detection systems... practices, and proper installation of all components (e.g., pipes) according to design specifications; (iii...

  13. 40 CFR 264.113 - Closure; time allowed for closure.

    Science.gov (United States)

    2010-07-01

    ... (CONTINUED) STANDARDS FOR OWNERS AND OPERATORS OF HAZARDOUS WASTE TREATMENT, STORAGE, AND DISPOSAL FACILITIES..., and closure and post-closure plans, and updated cost estimates and demonstrations of financial..., and removing all hazardous sludges to the extent practicable without impairing the integrity of the...

  14. Evidence for lipopolysaccharide-induced differentiation of RAW264 ...

    Indian Academy of Sciences (India)

    Unknown

    Beutler B 2000 Endotoxin, toll-like receptor 4, and the afferent limb of innate immunity; Curr. Opinion Microbiol. 3 23–28. Fenton M J and Golenbock D T 1998 LPS-binding proteins and receptors; J. Leukoc. Biol. 64 25–32. Grabbe S, Kampgen E and Schuler G 2000 Dendritic cells: multi-lineal and multi-functional; Immunol.

  15. Modelling of H.264 MPEG2 TS Traffic Source

    Directory of Open Access Journals (Sweden)

    Stanislav Klucik

    2013-01-01

    Full Text Available This paper deals with IPTV traffic source modelling. Traffic sources are used for simulation, emulation and real network testing. This model is made as a derivation of known recorded traffic sources that are analysed and statistically processed. As the results show the proposed model causes in comparison to the known traffic source very similar network traffic parameters when used in a simulated network.

  16. 40 CFR 264.573 - Design and operating requirements.

    Science.gov (United States)

    2010-07-01

    ..., climatic conditions, the stress of daily operations, e.g., variable and moving loads such as vehicle... leakage to which they are exposed, climatic conditions, the stress of installation, and the stress of..., construct, operate and maintain a run-on control system capable of preventing flow onto the drip pad during...

  17. 28 CFR 2.64 - Youth Corrections Act.

    Science.gov (United States)

    2010-07-01

    ... member who is knowledgeable about the case the program plan and the importance of good conduct and... has personal knowledge of the case shall be present to assist the examiners in their evaluation of the... to obey the law and favorably modified his value system. Participation in these programs will...

  18. Dicty_cDB: SHH264 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available sertion targeting and chromosome engineering clone MHPP142o16. 50 0.13 1 CR023465 |CR023465.1 Forward strand... read from insert in 3'HPRT insertion targeting and chromosome engineering clone MHPP421e10. 50 0.13 1 CR044...130 |CR044130.1 Forward strand read from insert in 3'HPRT insertion targeting and chromosome engineering clo

  19. 40 CFR 264.273 - Design and operating requirements.

    Science.gov (United States)

    2010-07-01

    ... the facility permit how the owner or operator will design, construct, operate, and maintain the land..., construct, operate, and maintain the treatment zone to minimize run-off of hazardous constituents during the... maintain a run-on control system capable of preventing flow onto the treatment zone during peak discharge...

  20. 40 CFR 264.151 - Wording of the instruments.

    Science.gov (United States)

    2010-07-01

    ... the State of ______” or “a national bank”], the “Trustee.” Whereas, the United States Environmental... qualified central depositary even though, when so deposited, such securities may be merged and held in bulk..., or any agency or instrumentality thereof, with a Federal Reserve bank, but the books and records of...