Science.gov

Sample records for 3t siemens trio

  1. Trios

    2011-09-29

    The Trilinos I/O support (Trios) software provides general I/O support for applications using Trilinos libraries. In particular, this software will include parallel I/O support to read and write selected Trilinos data structures to commonly-used file formats such as netCDF, hdf5, and Exodus.

  2. TRIO experiment

    SciTech Connect

    Clemmer, R.G.; Finn, P.A.; Malecha, R.F.; Misra, B.; Billone, M.C.; Bowers, D.L.; Fischer, A.K.; Greenwood, L.R.; Mattas, R.F.; Tam, S.W.

    1984-09-01

    The TRIO experiment is a test of in-situ tritium recovery and heat transfer performance of a miniaturized solid breeder blanket assembly. The assembly (capsule) was monitored for temperature and neutron flux profiles during irradiation and a sweep gas flowed through the capsule to an anaytical train wherein the amounts of tritium in its various chemical forms were determined. The capsule was designed to operate at different temperatures and sweep gas conditions. At the end of the experiment the amount of tritium retained in the solid was at a concentration of less than 0.1 wppM. More than 99.9% of tritium generated during the experiment was successfully recovered. The results of the experiment showed that the tritium inventories at the beginning and at the end of the experiment follow a relationship which appears to be characteristic of intragranular diffusion.

  3. Multimodal imaging: Simultaneous EEG in a 3T Hybrid MR-PET system

    NASA Astrophysics Data System (ADS)

    Neuner, I.; Warbrick, T.; Tellmann, L.; Rota Kops, E.; Arrubla, J.; Boers, F.; Herzog, H.; Shah, N. J.

    2013-02-01

    The new generation of integrated MR-PET systems allows the simultaneous acquisition of MR and PET data. While MR delivers structural data with an excellent spatial resolution, the advantage of PET is its information on a molecular level. However, both modalities have a low temporal resolution. Thus, for pharmacological studies or patients who suffer from treatment resistant epilepsy the combination of yet another modality such as EEG could be desirable. We tested the feasibility of evoked visual potentials in a 3T Hybrid MR-PET system (Siemens Germany) in comparison to a standalone 3T Trio System (Siemens Germany). A T2*-weighted EPI sequence was used: TR: 2.2 s, TE: 30 ms, FOV: 200 mm, slice thickness 3, 36 slices in a healthy volunteer (male, 27 years old) using an MR-compatible 32-channel EEG system (Brainproducts, Munich, Germany). We applied 200 trials of visual stimulation from a white and black checkerboard. Visual evoked potentials were analyzed using Brain Vision Analyzer (Brainproducts, Munich, Germany). Gradient correction and cardioballistic artefact correction were performed as implemented in Vision Analyzer. Visual event related potentials were successfully recorded at the 3T Hybrid MR-PET system. Both curves differ slightly in shape and latency due to the following factors: the distance from the screen varies slightly and the size of the field of view of the subjects is smaller in the 3T MR-PET system in comparison to the 3T stand alone system. Extending the 3T MR-PET Hybrid system to 3T Hybrid MR-PET-EEG is feasible and adds another tool to clinical neuroimaging and research.

  4. TRIO-01 experiment

    SciTech Connect

    Clemmer, R.G.; Finn, P.A.; Billone, M.C.; Misra, B.; Greenwood, L.R.; Dyer, F.F.; Dudley, I.T.; Bate, L.C.; Clemmer, E.D.; Fisher, P.W.

    1984-02-10

    The TRIO-01 experiment was designed to test in-situ tritium recovery and heat transfer performance of a candidate solid breeder, ..gamma..-LiAlO/sub 2/. The results showed that nearly all the tritium generated was recovered. Only <0.1 wppM tritium remained in the solid after irradiation testing. The heat transfer performance showed that temperature profiles can be effectively controlled.

  5. Trilinos I/O Support (Trios)

    DOE PAGESBeta

    Oldfield, Ron A.; Sjaardema, Gregory D.; Lofstead II, Gerald F.; Kordenbrock, Todd

    2012-01-01

    Trilinos I/O Support (Trios) is a new capability area in Trilinos that serves two important roles: (1) it provides and supports I/O libraries used by in-production scientific codes; (2) it provides a research vehicle for the evaluation and distribution of new techniques to improve I/O on advanced platforms. This paper provides a brief overview of the production-grade I/O libraries in Trios as well as some of the ongoing research efforts that contribute to the experimental libraries in Trios.

  6. New Expedition 32 Trio Arrives at Station

    NASA Video Gallery

    Expedition 32 Flight Engineers Suni Williams, Yuri Malenchenko and Aki Hoshide have arrived at the International Space Station after two days in orbit. The new trio docked its Soyuz TMA-05M spacecr...

  7. Trio gene is required for mouse learning ability.

    PubMed

    Zong, Wen; Liu, Shuoyang; Wang, Xiaotong; Zhang, Jian; Zhang, Tingting; Liu, Ziyi; Wang, Dongdong; Zhang, Aizhen; Zhu, Minsheng; Gao, Jiangang

    2015-05-22

    Trio is a guanine nucleotide exchange factor with multiple guanine nucleotide exchange factor domains. Trio regulates cytoskeleton dynamics and actin remodeling and is involved in cell migration and axonal guidance in neuronal development. The null allele of the Trio gene led to embryonic lethality, and Trio null embryos displayed aberrant organization in several regions of the brain at E18.5, including hippocampus. Nestin-Trio-/- mice, in which the Trio gene was deleted specifically in the neuronal system by the Nestin-Cre system, displayed severe phenotypes, including low survival rate, ataxia and multiple developmental defects of the cerebellum. All Nestin-Trio-/- mice died before reaching adulthood, which hinders research on Trio gene function in adult mice. Thus, we generated EMX1-Trio-/- mice by crossing Trio-floxed mice with EMX1-Cre mice in which Cre is expressed in the brain cortex and hippocampus. EMX1-Trio-/- mice can survive to adulthood. Trio gene deletion results in smaller brains, an abnormal hippocampus and disordered granule cells in the dentate gyrus (DG) and cornu ammonis (CA). Behavior tests showed that Trio deletion interfered with the hippocampal-dependent spatial learning in the mice, suggesting that Trio plays critical roles in the learning ability of adult mice. We conclude that the Trio gene regulates the neuronal development of the hippocampus and that it affects the intelligence of adult mice. PMID:25727174

  8. TRIO: Turbulent Response in Oxygen

    NASA Technical Reports Server (NTRS)

    Hecht, J. H.; Clemmons, J. H.; Katz, N.; Ben-Ami, M.; Carranza, P. A.; Larsen, M.

    2005-01-01

    This project was designed to build on the results from the successful launch of the Turbulent Oxygen Mixing Experiment (TOMEX) mother-daughter (instrumented and chemical-release) payload (21.126) that was launched in October 2000 from the White Sands Missile Range. The overall science objective was to investigate the evolution of the atmospheric response. at altitudes between 80 and 120 km, to the presence of unstable regions with vertical scales of the order of 1 to 10 km. TRIO was designed to use Na lidar measurements from the MAUUMALT observation on MAUI with a launch of a payload from Pacific Missile Range Facility (PMRF), located on Kauai. During this project, Aerospace participated in a Mission Initiation Conference. put together a science requirements document. performed a site visit to PMRF. prepared a CDR document. and developed a production and calibration procedure for one of the payload instruments. the 3-channel photometer. Unfortunately. NASA decided to terminate the program because of unforeseen (by NASA) range costs. This CDR document represents our view of this project at termination and provides a roadmap to perform this experiment should it be proposed again.

  9. TRIO (Triplet Ionospheric Observatory) Mission

    NASA Astrophysics Data System (ADS)

    Lee, D.; Seon, J.; Jin, H.; Kim, K.; Lee, J.; Jang, M.; Pak, S.; Kim, K.; Lin, R. P.; Parks, G. K.; Halekas, J. S.; Larson, D. E.; Eastwood, J. P.; Roelof, E. C.; Horbury, T. S.

    2009-12-01

    Triplets of identical cubesats will be built to carry out the following scientific objectives: i) multi-observations of ionospheric ENA (Energetic Neutral Atom) imaging, ii) ionospheric signature of suprathermal electrons and ions associated with auroral acceleration as well as electron microbursts, and iii) complementary measurements of magnetic fields for particle data. Each satellite, a cubesat for ion, neutral, electron, and magnetic fields (CINEMA), is equipped with a suprathermal electron, ion, neutral (STEIN) instrument and a 3-axis magnetometer of magnetoresistive sensors. TRIO is developed by three institutes: i) two CINEMA by Kyung Hee University (KHU) under the WCU program, ii) one CINEMA by UC Berkeley under the NSF support, and iii) three magnetometers by Imperial College, respectively. Multi-spacecraft observations in the STEIN instruments will provide i) stereo ENA imaging with a wide angle in local times, which are sensitive to the evolution of ring current phase space distributions, ii) suprathermal electron measurements with narrow spacings, which reveal the differential signature of accelerated electrons driven by Alfven waves and/or double layer formation in the ionosphere between the acceleration region and the aurora, and iii) suprathermal ion precipitation when the storm-time ring current appears. In addition, multi-spacecraft magnetic field measurements in low earth orbits will allow the tracking of the phase fronts of ULF waves, FTEs, and quasi-periodic reconnection events between ground-based magnetometer data and upstream satellite data.

  10. Supervillin Binds the Rac/Rho-GEF Trio and Increases Trio-mediated Rac1 Activation

    PubMed Central

    Son, Kyonghee; Smith, Tara C.; Luna, Elizabeth J.

    2015-01-01

    We investigated cross-talk between the membrane-associated, myosin II-regulatory protein supervillin and the actin-regulatory small GTPases Rac1, RhoA, and Cdc42. Supervillin knockdown reduced Rac1-GTP loading, but not the GTP loading of RhoA or Cdc42, in HeLa cells with normal levels of the Rac1-activating protein Trio. No reduction in Rac1-GTP loading was observed when supervillin levels were reduced in Trio-depleted cells. Conversely, overexpression of supervillin isoform 1 (SV1) or, especially, isoform 4 (SV4) increased Rac1 activation. Inhibition of the Trio-mediated Rac1 guanine nucleotide exchange (GEF) activity with ITX3 partially blocked the SV4-mediated increase in Rac1-GTP. Both SV4 and SV1 co-localized with Trio at or near the plasma membrane in ruffles and cell surface projections. Two sequences within supervillin bound directly to Trio spectrin repeats 4–7: SV1-171, which contains N-terminal residues found in both SV1 and SV4 and the SV4-specific differentially spliced coding exons 3, 4, and 5 within SV4 (SV4-E345; SV4 amino acids 276 – 669). In addition, SV4-E345 interacted with the homologous sequence in rat kalirin (repeats 4–7, amino acids 531 – 1101). Overexpressed SV1-174 and SV4-E345 affected Rac1-GTP loading, but only in cells with endogenous levels of Trio. Trio residues 771 – 1057, which contain both supervillin-interaction sites, exerted a dominant-negative effect on cell spreading. Supervillin and Trio knockdowns, separately or together, inhibited cell spreading, suggesting that supervillin regulates the Rac1 guanine nucleotide exchange activity of Trio, and potentially also kalirin, during cell spreading and lamellipodia extension. PMID:25655724

  11. Siemens and siemens-like processes for producing photovoltaics: Energy payback time and lifetime carbon emissions

    NASA Astrophysics Data System (ADS)

    Jiao, Yubo; Salce, Alex; Ben, Wade; Jiang, Feng; Ji, Xiaoyang; Morey, Evan; Lynch, David

    2011-01-01

    Polysilicon photovoltaics will play a significant role in meeting the world's shortfall in electrical energy this century. The photovoltaic industry relies on high-purity silicon produced in the Siemens process. New Siemens-like processes (which convert metallurgical silicon to trichlorosilane and deposit purified silicon through the decomposition of silane) and metallurgical processes for producing solar silicon are under development. Their energy payback time and lifetime carbon emissions are reviewed. The history of development of Siemens and Siemens-like processes is summarized.

  12. New Siemens Research Turbine - time lapse

    SciTech Connect

    2009-01-01

    The National Renewable Energy Laboratory (NREL) and Siemens Energy Inc. recently commissioned a new 2.3 megawatt Siemens wind turbine at NREL's National Wind Technology Center. This video shows a time lapse of the installation. The turbine is the centerpiece of a multi-year project to study the performance and aerodynamics of a new class of large, land-based machines — in what will be the biggest government-industry research partnership for wind power generation ever undertaken in the U.S.

  13. The system design of TRIO cinema Mission

    NASA Astrophysics Data System (ADS)

    Jin, Ho; Seon, Jongho; Kim, Khan-Hyuk; Lee, Dong-Hun; Kim, Kap-Sung; Lin, Robert; Parks, George; Tindall, Craig; Horbury, T. S.; Larson, Davin; Sample, John

    TRIO (Triplet Ionospheric Observatory) CINEMA ( Cubesat for Ion, Neutral, Electron, MAg-netic fields) is a space science mission with three identical cubesats. The main scientific objec-tives are a multi-observation of ionospheric ENA (Energetic Neutral Atom) imaging, ionospheric signature of suprathermal electrons and ions and complementary measurements of magnetic fields for particle data. For this, Main payloads consist of a suprathermal electron, ion, neutral (STEIN) instrument and a 3-axis magnetometer of magnetoresistive sensors. The CINEMA is a 3-unit CubeSat, which translates to a 10 cm x 10 cm x 30 cm in volume and no more than four kilograms in mass. An attitude control system (ACS) uses torque coils, a sun sensor and the magnetometers and spin CINEMA spcaecraft 4 rpm with the spin axis perpendicular to the ecliptic plane. CINEMA will be placed into a high inclination low earth orbit that crosses the auroral zone and cusp. Three institutes are collaborating to develop CINEMA cubesats: i) two cubesats by Kyung Hee University (KHU) under their World Class University (WCU) program, ii) one cubesat by UC Berkeley under the NSF support, and iii) three magnetometers are provide by Imperial College, respectively. In this paper, we describe the system design and their performance of TR IO cinema mission. TRIO cinema's development of miniature in-strument and spacecraft spinning operation will play an important role for future nanosatellite space missions

  14. Siemens Foundation and the STEM Challenge

    ERIC Educational Resources Information Center

    Harper-Taylor, Jeniffer

    2010-01-01

    For more than 12 years, the Siemens Foundation has found unique ways to partner with organizations to support educational initiatives in science, technology, engineering and mathematics (STEM) in the United States. Its focus is clear--to educate the next generation of innovators by supporting math and science education from grade school to grad…

  15. 34 CFR 642.1 - Training Program for Federal TRIO Programs.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 34 Education 3 2010-07-01 2010-07-01 false Training Program for Federal TRIO Programs. 642.1... POSTSECONDARY EDUCATION, DEPARTMENT OF EDUCATION TRAINING PROGRAM FOR FEDERAL TRIO PROGRAMS General § 642.1 Training Program for Federal TRIO Programs. The Training Program for Federal TRIO Programs—referred to...

  16. Abdominal adipose tissue quantification on water-suppressed and non-water-suppressed MRI at 3T using semi-automated FCM clustering algorithm

    NASA Astrophysics Data System (ADS)

    Valaparla, Sunil K.; Peng, Qi; Gao, Feng; Clarke, Geoffrey D.

    2014-03-01

    Accurate measurements of human body fat distribution are desirable because excessive body fat is associated with impaired insulin sensitivity, type 2 diabetes mellitus (T2DM) and cardiovascular disease. In this study, we hypothesized that the performance of water suppressed (WS) MRI is superior to non-water suppressed (NWS) MRI for volumetric assessment of abdominal subcutaneous (SAT), intramuscular (IMAT), visceral (VAT), and total (TAT) adipose tissues. We acquired T1-weighted images on a 3T MRI system (TIM Trio, Siemens), which was analyzed using semi-automated segmentation software that employs a fuzzy c-means (FCM) clustering algorithm. Sixteen contiguous axial slices, centered at the L4-L5 level of the abdomen, were acquired in eight T2DM subjects with water suppression (WS) and without (NWS). Histograms from WS images show improved separation of non-fatty tissue pixels from fatty tissue pixels, compared to NWS images. Paired t-tests of WS versus NWS showed a statistically significant lower volume of lipid in the WS images for VAT (145.3 cc less, p=0.006) and IMAT (305 cc less, p<0.001), but not SAT (14.1 cc more, NS). WS measurements of TAT also resulted in lower fat volumes (436.1 cc less, p=0.002). There is strong correlation between WS and NWS quantification methods for SAT measurements (r=0.999), but poorer correlation for VAT studies (r=0.845). These results suggest that NWS pulse sequences may overestimate adipose tissue volumes and that WS pulse sequences are more desirable due to the higher contrast generated between fatty and non-fatty tissues.

  17. A novel strategy for sibship determination in trio sibling model

    PubMed Central

    Lee, James Chun-I; Lin, Yen-Yang; Tsai, Li-Chin; Lin, Chun-Yen; Huang, Tsun-Ying; Chu, Pao-Ching; Yu, Yu-Jen; Linacre, Adrian; Hsieh, Hsing-Mei

    2012-01-01

    Aim To use a virtually simulated population, generated from published allele frequencies based on 15 short tandem repeats (STR), to evaluate the efficacy of trio sibship testing and sibling assignment for forensic purposes. Methods Virtual populations were generated using 15 STR loci to create a large number of related and unrelated genotypes (10 000 trio combinations). Using these virtual populations, the probability of related and unrelated profiles can be compared to determine the chance of inclusions of being siblings if they are true siblings and the chance of inclusion if they are unrelated. Two specific relationships were tested – two reference siblings were compared to a third true sibling (3S trio, sibling trio) and two reference siblings were compared to an unrelated individual (2S1U trio, non-sibling trio). Results When the likelihood ratio was greater than 1, 99.87% of siblings in the 3S trio population were considered as siblings (sensitivity); 99.88% of non-siblings in the 2S1U trio population were considered as non-siblings (specificity); 99.9% of both populations were identified correctly as siblings and non-siblings; and the accuracy of the test was 99.88%. Conclusions The high sensitivity and specificity figures when using two known siblings compared to a putative sibling are significantly greater than when using only one known relative. The data also support the use of increasing number of loci allowing for greater confidence in genetic identification. The system established in this study could be used as the model for evaluating and simulating the cases with multiple relatives. PMID:22911526

  18. Whole genome analysis of a Vietnamese trio.

    PubMed

    Hai, Dang Thanh; Thanh, Nguyen Dai; Trang, Pham Thi Minh; Quang, Le Si; Hang, Phan Thi Thu; Cuong, Dang Cao; Phuc, Hoang Kim; Duc, Nguyen Huu; Dong, Do Duc; Minh, Bui Quang; Son, Pham Bao; Vinh, Le Sy

    2015-03-01

    We here present the first whole genome analysis of an anonymous Kinh Vietnamese (KHV) trio whose genomes were deeply sequenced to 30-fold average coverage. The resulting short reads covered 99.91 percent of the human reference genome (GRCh37d5). We identified 4,719,412 SNPs and 827,385 short indels that satisfied the Mendelian inheritance law. Among them, 109,914 (2.3 percent) SNPs and 59,119 (7.1 percent) short indels were novel. We also detected 30,171 structural variants of which 27,604 (91.5 percent) were large indels. There were 6,681 large indels in the range 0.1-100 kbp occurring in the child genome that were also confirmed in either the father or mother genome. We compared these large indels against the DGV database and found that 1,499 (22.44 percent) were KHV specific. De novo assembly of high-quality unmapped reads yielded 789 contigs with the length greater than or equal to 300 bp. There were 235 contigs from the child genome of which 199 (84.7 percent) were significantly matched with at least one contig from the father or mother genome. Blasting these 199 contigs against other alternative human genomes revealed 4 novel contigs. The novel variants identified from our study demonstrated the necessity of conducting more genome-wide studies not only for Kinh but also for other ethnic groups in Vietnam. PMID:25740146

  19. 76 FR 59167 - Siemens Medical Solutions USA, Inc., Oncology Care Systems Division, Concord, CA; Siemens Medical...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-09-23

    ... Register on April 23, 2010 (75 FR 21355). The Department's Notice was amended on July 29, 2011 to include... the Federal Register on August 12, 2011 (76 FR 50269). The workers are engaged in employment related... Employment and Training Administration Siemens Medical Solutions USA, Inc., Oncology Care Systems...

  20. Status of SOFC development at Siemens

    SciTech Connect

    Drenckhahn, W.; Blum, L.; Greiner, H.

    1996-12-31

    The Siemens SOFC development programme reached an important milestone in June 1995. A stack operating with hydrogen and oxygen produced a peak power of 10.7 kW at a current density of 0.7 A/cm{sup 2} and was running for more than 1400 hours. The SOFC configuration is based on a flat metal separator plate using the multiple cell array design. Improved PENs, functional layer and joining technique were implemented. Based on this concept, a 100 kW plant was designed The SOFC development at Siemens has been started in 1990 after a two years preparation phase. The first period with the goal of the demonstration of a 1 kW SOFC stack operation ended in 1993. This important milestone was finally reached in the begin of 1994. The second project phase with the final milestone of a 20 kW module operation will terminate at the end of 1996. This result will form a basis for the next phase in which a 50 to 100 kW pilot plant will be built and tested.

  1. A Trio of Super-Earths

    NASA Astrophysics Data System (ADS)

    2008-06-01

    Today, at an international conference, a team of European astronomers announced a remarkable breakthrough in the field of extra-solar planets. Using the HARPS instrument at the ESO La Silla Observatory, they have found a triple system of super-Earths around the star HD 40307. Moreover, looking at their entire sample studied with HARPS, the astronomers count a total of 45 candidate planets with a mass below 30 Earth masses and an orbital period shorter than 50 days. This implies that one solar-like star out of three harbours such planets. A trio of Super-Earths ESO PR Photo 19a/08 A trio of Super-Earths "Does every single star harbour planets and, if yes, how many?" wonders planet hunter Michel Mayor from Geneva Observatory. "We may not yet know the answer but we are making huge progress towards it." Since the discovery in 1995 of a planet around the star 51 Pegasi by Mayor and Didier Queloz, more than 270 exoplanets have been found, mostly around solar-like stars. Most of these planets are giants, such as Jupiter or Saturn, and current statistics show that about 1 out of 14 stars harbours this kind of planet. "With the advent of much more precise instruments such as the HARPS spectrograph on ESO's 3.6-m telescope at La Silla, we can now discover smaller planets, with masses between 2 and 10 times the Earth's mass," says Stéphane Udry, one of Mayor's colleagues. Such planets are called super-Earths, as they are more massive than the Earth but less massive than Uranus and Neptune (about 15 Earth masses). The group of astronomers have now discovered a system of three super-Earths around a rather normal star, which is slightly less massive than our Sun, and is located 42 light-years away towards the southern Doradus and Pictor constellations. "We have made very precise measurements of the velocity of the star HD 40307 over the last five years, which clearly reveal the presence of three planets," says Mayor. The planets, having 4.2, 6.7, and 9.4 times the mass of the

  2. TRIO/Special Services Program Evaluation. Final Report, 1980-81.

    ERIC Educational Resources Information Center

    Read, Sherry

    The TRIO/Special Services Program at the University of Minnesota is described, and a national evaluation of TRIO programs (Upward Bound, Talent Search, and Special Services) is briefly summarized. The TRIO/Special Services Program at the University of Minnesota is located within The General College, which is the open enrollment unit of the…

  3. 75 FR 65711 - High School Equivalency Program and College Assistance Migrant Program, The Federal TRIO Programs...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-10-26

    ...The Secretary amends the regulations, and establishes new regulations, for the High School Equivalency Program and College Assistance Migrant Program (HEP and CAMP); the Federal TRIO programs (TRIO programs--Training program for Federal TRIO programs (Training), Talent Search (TS), Educational Opportunity Centers (EOC), Upward Bound (UB), Student Support Services (SSS), and the Ronald E.......

  4. A Tutor Handbook for TRIO Programs. Operation Success.

    ERIC Educational Resources Information Center

    de Silva, Deema; Freund, Elizabeth

    This handbook for tutors participating in the federally funded Operation Success Special Services Program, one of the college level TRIO programs designed to aid students from disadvantaged backgrounds, was compiled with the cooperation of tutors at the Wichita State University. The first section describes the needs of the students being tutored…

  5. Unique and Effective Practices for TRIO Student Support Services Programs.

    ERIC Educational Resources Information Center

    Walsh, Julia

    The paper discusses the various services provided through the federally funded TRIO Student Support Services (SSS) program at Kankakee Community College (KCC) (Illinois). The program helps simplify the educational process for low-income, first-generation and disabled college students. The goal is to ensure qualified students get the support they…

  6. Financial Education in TRIO Programs. Institutional Policy Brief

    ERIC Educational Resources Information Center

    Yang, Hannah; Kezar, Adrianna

    2009-01-01

    To address some of the financial challenges facing low-income students, federal policymakers enacted a provision in the 2008 Higher Education Opportunity Act (HEOA) that makes financial literacy a required service of all TRIO programs (or, in the case of McNair, simply makes permissible). Effective August 2008, these programs started offering…

  7. Evaluation of the Siemens Minifonator vibrotactile aid.

    PubMed

    Weisenberger, J M

    1989-03-01

    The Siemens Hearing Instruments Minifonator, a single-channel, wrist-worn vibrotactile aid, was evaluated in a laboratory setting with hearing-impaired adults. Eight subjects, with hearing loss greater than 70 dB in the better ear, were administered a test battery including sound-field detection, speech awareness threshold, environmental sound identification, syllable rhythm and stress categorization, and sentence identification subtests. Performance on each subtest was compared for hearing-aided and tactile-aided conditions. Mean performance levels for the two conditions were comparable for all but one subtest. However, closer inspection revealed that the data for the hearing-aided condition were bimodal: some subjects proved to be good hearing aid users and other performed at chance levels with their hearing aids. Performance in the tactile-aided conditions did not show such variability and mean levels fell between the hearing-aided levels for "good" and "poor" hearing aid users. The effects of training with the device were assessed for two normal-hearing subjects, who were trained in 1-hour daily sessions over a several week period on the environmental sounds and syllable rhythm and stress subtests. Results indicated substantial improvements in performance over the course of training. The implications of these results for long-term use of the device are discussed. PMID:2704198

  8. Focal Cortical Lesion Detection in Multiple Sclerosis: 3T DIR versus 7T FLASH-T2*

    PubMed Central

    Nielsen, A. Scott; Kinkel, R. Philip; Tinelli, Emanuele; Benner, Thomas; Cohen-Adad, Julien; Mainero, Caterina

    2014-01-01

    Purpose To evaluate the inter-rater agreement of cortical lesion detection using 7T FLASH-T2* and 3T DIR sequences. Materials and Methods Twenty-six patients with multiple sclerosis were scanned on a human 7T (Sidemen’s) and 3T MRI (TIM Trio, Sidemen’s) to acquire 3T DIR/MEMPR and 7T FLASH-T2* sequences. Four independent reviewers scored and categorized cortical lesions in the bilateral pre-central gyri (motor strips) as leukocortical, intracortical, or subpial. Inter-rater agreement was assessed according to lesion category using the kappa statistic. The sensitivity of recent MAGNIMS consensus guidelines for cortical lesion detection using 3T DIR was assessed with 7T FLASH-T2* as the reference gold standard. Results Inter-rater agreement at 7T was excellent compared to 3T (k=0.97 vs. 0.12). FLASH-T2* at 7T detected subpial lesions while 3T DIR did not. The predicted sensitivity of 3T DIR sequence for cortical lesions in vivo is modest (range of 13.6 to 18.3%). Conclusion 7T FLASH-T2* detects more cortical—particularly subpial—lesions compared to 3T DIR. In the absence of DIR/post-mortem data, 7T FLASH-T2* is a suitable gold-standard instrument and should be incorporated into future consensus guidelines. PMID:22045554

  9. TRIO-01 experiment: in-situ tritium-recovery results

    SciTech Connect

    Clemmer, R.G.; Finn, P.A.; Billone, M.C.; Misra, B.; Arons, R.M.; Poeppel, R.B.; Dyer, F.F.; Dudley, I.T.; Bate, L.C.; Clemmer, E.D.

    1983-08-01

    The TRIO-01 experiment is a test of in-situ tritium recovery from ..gamma..-LiAlO/sub 2/ with test conditions chosen to simulate those anticipated in fusion power reactors. A status report is presented which describes qualitatively the results observed during the irradiation phase of the experiment. Both the rate of tritium release and the chemical forms of tritium were measured using a helium sweep gas which flowed past the breeder material to a gas analysis system.

  10. TRIO-01 experiment: in-situ tritium recovery results

    SciTech Connect

    Clemmer, R.G.; Finn, P.A.; Billone, M.C.; Misra, B.; Arons, R.M.; Poeppel, R.B.; Dyer, F.F.; Dudley, I.T.; Bate, L.C.; Clemmer, E.D.

    1983-10-01

    The TRIO-01 experiment is a test of in-situ tritium recovery from ..gamma..-LiAlO/sub 2/ with test conditions chosen to simulate those anticipated in fusion power reactors. A status report is presented which describes qualitatively the results observed during the irradiation phase of the experiment. Both the rate of tritium release and the chemical forms of tritium were measured using a helium sweep gas which flowed past the breeder material to a gas analysis system.

  11. TRIO-01 experiment: in-situ tritium recovery results

    SciTech Connect

    Clemmer, R.G.; Finn, P.A.; Billone, M.C.; Misra, B.; Greenwood, L.R.; Dyer, F.F.; Dudley, I.T.; Bate, L.C.; Clemmer, E.D.; Fisher, P.W.

    1983-01-01

    The TRIO-01 experiment was designed to test in-situ tritium recovery and heat transfer performance of a candidate solid breeder, ..gamma..-LiAlO/sub 2/. The results showed that nearly all the tritium generated was recovered. Only < 0.1 wppM tritium remained in the solid after irradiation testing. The heat transfer performance showed that temperature profiles can be effectively controlled.

  12. TRIO: Burst Buffer Based I/O Orchestration

    SciTech Connect

    Wang, Teng; Oral, H Sarp; Pritchard, Michael; Wang, Bin; Yu, Weikuan

    2015-01-01

    The growing computing power on leadership HPC systems is often accompanied by ever-escalating failure rates. Checkpointing is a common defensive mechanism used by scientific applications for failure recovery. However, directly writing the large and bursty checkpointing dataset to parallel filesystem can incur significant I/O contention on storage servers. Such contention in turn degrades the raw bandwidth utilization of storage servers and prolongs the average job I/O time of concurrent applications. Recently burst buffer has been proposed as an intermediate layer to absorb the bursty I/O traffic from compute nodes to storage backend. But an I/O orchestration mechanism is still desired to efficiently move checkpointing data from bursty buffers to storage backend. In this paper, we propose a burst buffer based I/O orchestration framework, named TRIO, to intercept and reshape the bursty writes for better sequential write traffic to storage severs. Meanwhile, TRIO coordinates the flushing orders among concurrent burst buffers to alleviate the contention on storage server bandwidth. Our experimental results reveal that TRIO can deliver 30.5% higher bandwidth and reduce the average job I/O time by 37% on average for data-intensive applications in various checkpointing scenarios.

  13. Privacy Impact Assessment for the TRIO Programs Annual Performance Report (APR) System

    ERIC Educational Resources Information Center

    US Department of Education, 2008

    2008-01-01

    The TRIO Programs Annual Performance Report (APR) System collects individual student records on individuals served by the following Federal TRIO Programs: Upward Bound (which includes regular Upward Bound (UB), Upward Bound Math-Science (UBMS), and Veterans Upward Bound (VUB)); Student Support Services (SSS); and the Ronald E. McNair Post…

  14. National Studies Find TRIO Programs Effective at Increasing College Enrollment and Graduation

    ERIC Educational Resources Information Center

    Pell Institute for the Study of Opportunity in Higher Education, 2009

    2009-01-01

    Since 1997, four national studies conducted by the U.S. Department of Education have been completed on the Federal TRIO programs. Data from these studies demonstrate that participation in a TRIO program has a significant impact on the educational outcomes of low-income, first-generation students and students with disabilities. This paper provides…

  15. 77 FR 28360 - Applications for New Awards; Training Program for Federal TRIO Programs

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-05-14

    ... From the Federal Register Online via the Government Publishing Office DEPARTMENT OF EDUCATION Applications for New Awards; Training Program for Federal TRIO Programs AGENCY: Office of Postsecondary Education, Department of Education. ACTION: Notice. Overview Information Training Program for Federal TRIO Programs (Training Program)....

  16. The Integrated Course of Study in the General College TRIO Program.

    ERIC Educational Resources Information Center

    Hixson, Bruce, Ed.

    1982-01-01

    The TRIO program at the General College, University of Minnesota, is addressed in this newsletter issue, based on presentations made at the 1981 annual meeting of the Mid-American Association of Educational Opportunity Program Personnel. The name TRIO stands for three federal programs: (1) Upward Bound, which assists high school students from…

  17. 3T MR imaging of the brain.

    PubMed

    DeLano, Mark C; Fisher, Charles

    2006-02-01

    The advent of very high field clinical scanners that operate at 3T is taking structural and functional imaging to new levels and is reinvigorating clinical spectroscopy, fMR imaging, and noncontrast-enhanced methods of MRA. Most of the challenges that are related to 3T imaging have been addressed to facilitate routine clinical imaging. An awareness of the complexities that underlie the solutions to these challenges is important to the continued improvements to the 3T platform so that its maximal potential can be reached. The development of the multichannel-head coils and the improvement in the design of body coils, concurrently with the development of multichannel capabilities that enable parallel imaging, have benefited all field platforms. Perhaps the added value of parallel imaging has been greatest at 3T where the additional signal can be exploited. The definition of very high field is a moving target, and may be well on its way to 7.0 T, although in terms of the current clinical state of the art, 3T is our current reference. PMID:16530636

  18. New Expedition 35 Trio Checks out Soyuz - Duration: 6 minutes, 38 seconds.

    NASA Video Gallery

    The newest Expedition 35 trio is in Kazakhstan preparing for its March 28 launch and docking to the International Space Station. Ground personnel at the Baikonur Cosmodrome are readying the Soyuz T...

  19. Index of Human Well-Being for the U.S.: A TRIO Approach

    EPA Science Inventory

    In 2011, the US Environmental Protection Agency’s (EPA) Office of Research and Development program, Sustainable and Healthy Communities, coined the term TRIO (Total Resources Impact Outcome) to represent approaches that fully incorporate all three pillars of community susta...

  20. An Index of Human Well-being for the U.S.: A TRIO Approach

    EPA Science Inventory

    : In 2011, EPA’s Office of Research and Development program, Sustainable and Healthy Communities, coined the term TRIO (Total Resources Impact Outcome) to represent approaches the fully incorporate all three pillars of community sustainability – environmental, economic and social...

  1. Optimal clinical implementation of the Siemens virtual wedge

    SciTech Connect

    Walker, C.P.; Richmond, N.D.; Lambert, G.D

    2003-09-30

    Installation of a modern high-energy Siemens Primus linear accelerator at the Northern Centre for Cancer Treatment (NCCT) provided the opportunity to investigate the optimal clinical implementation of the Siemens virtual wedge filter. Previously published work has concentrated on the production of virtual wedge angles at 15 deg., 30 deg., 45 deg., and 60 deg. as replacements for the Siemens hard wedges of the same nominal angles. However, treatment plan optimization of the dose distribution can be achieved with the Primus, as its control software permits the selection of any virtual wedge angle from 15 degree sign to 60 degree sign in increments of 1 deg. The same result can also be produced from a combination of open and 60 deg. wedged fields. Helax-TMS models both of these modes of virtual wedge delivery by the wedge angle and the wedge fraction methods respectively. This paper describes results of timing studies in the planning of optimized patient dose distributions by both methods and in the subsequent treatment delivery procedures. Employment of the wedge fraction method results in the delivery of small numbers of monitor units to the beam's central axis; therefore, wedge profile stability and delivered dose with low numbers of monitor units were also investigated. The wedge fraction was proven to be the most efficient method when the time taken for both planning and treatment delivery were taken into consideration, and is now used exclusively for virtual wedge treatment delivery in Newcastle. It has also been shown that there are no unfavorable dosimetric consequences from its practical implementation.

  2. Seeman-Bohlin linkage for Siemens x-ray diffractometer

    NASA Astrophysics Data System (ADS)

    Banerjee, R. L.; Richard, Aldérice

    1991-04-01

    A mechanical linkage attached to a Siemens x-ray diffractometer is designed and constructed to convert it to a Seeman-Bohlin diffractometer, with special emphasis on its suitability for x-ray characterization of polycrystalline thin films. The apparatus employs a pyrolitic graphite monochromator crystal using the full focusing Johansson principle for the diffracted beam to obtain a high intensity low background diffraction pattern. The performance of this Seeman-Bohlin diffractometer is illustrated by recording the diffraction pattern of a 100-Å-thick thin film of gold.

  3. Christ siemens touraine syndrome: A rare case report.

    PubMed

    Retnakumari, N; Varghese, Manuja; Kannan, V P

    2016-01-01

    Christ-Siemens-Touraine (CST) is a rare hereditary disorder of X-linked recessive trait, characterized by abnormal development of two or more structures or tissues of ectodermal origin. The common clinical findings include hypodontia, hypohydrosis, hypotrichosis, and onychodysplasia. Although hypodontia is common, anodontia is a rare feature. Most of the patients are suffering from social rejection and consequent psychological trauma because of the facial dysmorphism and absence of multiple teeth. Oral rehabilitation is of prime importance for such patients. This article presents a case in a 5½-year-old boy presenting with altered manifestations affecting almost all the ectodermal structures like skin, hair, nails, teeth, sebaceous glands, sweat glands, salivary glands, mammary glands, and tear glands. He also had complete anodontia and dry mouth. A multidisciplinary treatment was given to the patient with the collaboration of various health professionals. The child gained confidence and was relieved from the psychological impact following the prosthetic rehabilitation. PMID:27080972

  4. Mexican American First-Generation/Low-Income Students: A Rural Community College, TRiO Student Support Services Experience

    ERIC Educational Resources Information Center

    O'Meara, Daniel J.

    2012-01-01

    This study is an ethnographic inquiry into the beliefs and perceptions of first-generation/low-income Mexican American students in a rural community college located near the U.S.-Mexico border. It explored their experiences as TRiO Student Support Services participants. TRiO Student Support Services plays an increasingly vital role helping…

  5. 77 FR 31390 - Manufacturer of Controlled Substances; Notice of Registration; Siemens Healthcare Diagnostics, Inc.

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-05-25

    ..., 77 FR 5847, Siemens Healthcare Diagnostics Inc., 100 GBC Drive, Mail Stop 514, Newark, Delaware 19702, made application by renewal to the Drug Enforcement Administration (DEA) to be registered as a bulk... Enforcement Administration Manufacturer of Controlled Substances; Notice of Registration; Siemens...

  6. 77 FR 72409 - Manufacturer of Controlled Substances; Notice of Application: Siemens Healthcare Diagnostics, Inc.

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-12-05

    ... Enforcement Administration Manufacturer of Controlled Substances; Notice of Application: Siemens Healthcare... notice that on November 7, 2012, Siemens Healthcare Diagnostics, Inc., Attn: RA, 100 GBC Drive, Mail Stop 514, Newark, Delaware 19702, made application by renewal to the Drug Enforcement Administration...

  7. ETV Report: Siemens Model V-40R-A150 Open Channel UV System

    EPA Science Inventory

    Verification testing of the Siemens Barrier Sunlight V-40R-A150 UV System was completed at the UV Validation and Research Center of New York (UV Center), located in Johnstown, NY. The V-40R System supplied by Siemens utilizes 40 high-output, low-pressure amalgam lamps, oriented ...

  8. The Rho-GEF Trio regulates a novel pro-inflammatory pathway through the transcription factor Ets2.

    PubMed

    Van Rijssel, Jos; Timmerman, Ilse; Van Alphen, Floris P J; Hoogenboezem, Mark; Korchynskyi, Olexandr; Geerts, Dirk; Geissler, Judy; Reedquist, Kris A; Niessen, Hans W M; Van Buul, Jaap D

    2013-06-15

    Inflammation is characterized by endothelium that highly expresses numerous adhesion molecules to trigger leukocyte extravasation. Central to this event is increased gene transcription. Small Rho-GTPases not only control the actin cytoskeleton, but are also implicated in gene regulation. However, in inflammation, it is not clear how this is regulated. Here, we show that the guanine-nucleotide exchange factor Trio expression is increased upon inflammatory stimuli in endothelium. Additionally, increased Trio expression was found in the vessel wall of rheumatoid arthritis patients. Trio silencing impaired VCAM-1 expression. Finally, we excluded that Trio-controlled VCAM-1 expression used the classical NFκB or MAP-kinase pathways, but rather acts on the transcriptional level by increasing phosphorylation and nuclear translocalization of Ets2. These data implicate Trio in regulating inflammation and provide novel targets for therapeutic purposes to treat inflammatory diseases such as rheumatoid arthritis. PMID:23789107

  9. Bootstraps: Federal Trio Programs, if Funded, Could Help Close Income Gap

    ERIC Educational Resources Information Center

    Jean, Reggie

    2011-01-01

    Since 1964, the federal government has had two successful programs that have helped Americans from low-income and first-generation college backgrounds (whose parents never enrolled in higher education) prepare for and earn their college degrees, helping to stop the cycle of poverty. The federally funded TRIO programs (Upward Bound, Veterans Upward…

  10. Increasing Underrepresented High School Students' College Transitions and Achievements: TRIO Educational Opportunity Programs

    ERIC Educational Resources Information Center

    Cowan Pitre, Charisse; Pitre, Paul

    2009-01-01

    The American education agenda suggests a clear commitment to the development of programs and practices to increase equitable participation in higher education. During a time when equity-based policy initiatives are under attack in the United States, governmental TRIO Programs remain one proven pathway for ensuring college preparedness and access…

  11. A Profile of the Federal TRIO Programs and Child Care Access Means Parents in School Program

    ERIC Educational Resources Information Center

    US Department of Education, 2008

    2008-01-01

    To help fulfill the goal of No Child Left Behind (NCLB), high-quality postsecondary education opportunities must be available to all students. In keeping with this goal, the Federal TRIO programs and Child Care Access Means Parents in School (CCAMPIS) program provide outreach and support to help low-income and first-generation college students, as…

  12. Analysis of Case-Parent Trios Using a Loglinear Model with Adjustment for Transmission Ratio Distortion

    PubMed Central

    Huang, Lam O.; Infante-Rivard, Claire; Labbe, Aurélie

    2016-01-01

    Transmission of the two parental alleles to offspring deviating from the Mendelian ratio is termed Transmission Ratio Distortion (TRD), occurs throughout gametic and embryonic development. TRD has been well-studied in animals, but remains largely unknown in humans. The Transmission Disequilibrium Test (TDT) was first proposed to test for association and linkage in case-trios (affected offspring and parents); adjusting for TRD using control-trios was recommended. However, the TDT does not provide risk parameter estimates for different genetic models. A loglinear model was later proposed to provide child and maternal relative risk (RR) estimates of disease, assuming Mendelian transmission. Results from our simulation study showed that case-trios RR estimates using this model are biased in the presence of TRD; power and Type 1 error are compromised. We propose an extended loglinear model adjusting for TRD. Under this extended model, RR estimates, power and Type 1 error are correctly restored. We applied this model to an intrauterine growth restriction dataset, and showed consistent results with a previous approach that adjusted for TRD using control-trios. Our findings suggested the need to adjust for TRD in avoiding spurious results. Documenting TRD in the population is therefore essential for the correct interpretation of genetic association studies.

  13. A News Game Called TRIO: A Task for Reporting, Interviewing and Observing.

    ERIC Educational Resources Information Center

    Talbott, Albert D.; And Others

    The reason for creating the Task for Reporting, Interviewing, and Observing (TRIO) was to make selective perception and metaphoric transformation come alive for students. This paper includes the experiences in designing, implementing, and trying out the exercise, a description of the exercise, a summary of the participants' play, and suggestions…

  14. Using Constructivist Career Development to Improve Career Decision Self-Efficacy in TRiO Students

    ERIC Educational Resources Information Center

    Grier-Reed, Tabitha; Ganuza, Zoila

    2012-01-01

    Although more high school graduates are attending college, many are not graduating (The Bill and Melinda Gates Foundation, 2004). First-generation, low-income, and underrepresented students are especially at risk for falling through the cracks. To help address this issue, programs such as TRiO Student Support Services (SSS) assist…

  15. 75 FR 37415 - Office of Postsecondary Education; Overview Information; Training Program for Federal TRIO...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-06-29

    ... Office of Postsecondary Education; Overview Information; Training Program for Federal TRIO Programs (Training Program) Notice inviting applications for new awards for fiscal year (FY) 2010. Catalog of Federal... Purpose of Program: The Training Program provides grants to train the staff and leadership...

  16. Fast detection of de novo copy number variants from SNP arrays for case-parent trios

    PubMed Central

    2012-01-01

    Background In studies of case-parent trios, we define copy number variants (CNVs) in the offspring that differ from the parental copy numbers as de novo and of interest for their potential functional role in disease. Among the leading array-based methods for discovery of de novo CNVs in case-parent trios is the joint hidden Markov model (HMM) implemented in the PennCNV software. However, the computational demands of the joint HMM are substantial and the extent to which false positive identifications occur in case-parent trios has not been well described. We evaluate these issues in a study of oral cleft case-parent trios. Results Our analysis of the oral cleft trios reveals that genomic waves represent a substantial source of false positive identifications in the joint HMM, despite a wave-correction implementation in PennCNV. In addition, the noise of low-level summaries of relative copy number (log R ratios) is strongly associated with batch and correlated with the frequency of de novo CNV calls. Exploiting the trio design, we propose a univariate statistic for relative copy number referred to as the minimum distance that can reduce technical variation from probe effects and genomic waves. We use circular binary segmentation to segment the minimum distance and maximum a posteriori estimation to infer de novo CNVs from the segmented genome. Compared to PennCNV on simulated data, MinimumDistance identifies fewer false positives on average and is comparable to PennCNV with respect to false negatives. Genomic waves contribute to discordance of PennCNV and MinimumDistance for high coverage de novo calls, while highly concordant calls on chromosome 22 were validated by quantitative PCR. Computationally, MinimumDistance provides a nearly 8-fold increase in speed relative to the joint HMM in a study of oral cleft trios. Conclusions Our results indicate that batch effects and genomic waves are important considerations for case-parent studies of de novo CNV, and that the

  17. Monte Carlo study of Siemens PRIMUS photoneutron production

    NASA Astrophysics Data System (ADS)

    Pena, J.; Franco, L.; Gómez, F.; Iglesias, A.; Pardo, J.; Pombar, M.

    2005-12-01

    Neutron production in radiotherapy facilities has been studied from the early days of modern linacs. Detailed studies are now possible using photoneutron capabilities of general-purpose Monte Carlo codes at energies of interest in medical physics. The present work studies the effects of modelling different accelerator head and room geometries on the neutron fluence and spectra predicted via Monte Carlo. The results from the simulation of a 15 MV Siemens PRIMUS linac show an 80% increase in the fluence scored at the isocentre when, besides modelling the components neccessary for electron/photon simulations, other massive accelerator head components are included. Neutron fluence dependence on inner treatment room volume is analysed showing that thermal neutrons have a 'gaseous' behaviour and then a 1/V dependence. Neutron fluence maps for three energy ranges, fast (E > 0.1 MeV), epithermal (1 eV < E < 0.1 MeV) and thermal (E < 1 eV), are also presented and the influence of the head components on them is discussed.

  18. Type testing of the Siemens Plessey electronic personal dosemeter.

    PubMed

    Hirning, C R; Yuen, P S

    1995-07-01

    This paper presents the results of a laboratory assessment of the performance of a new type of personal dosimeter, the Electronic Personal Dosemeter made by Siemens Plessey Controls Limited. Twenty pre-production dosimeters and a reader were purchased by Ontario Hydro for the assessment. Tests were performed on radiological performance, including reproducibility, accuracy, linearity, detection threshold, energy response, angular response, neutron response, and response time. There were also tests on the effects of a variety of environmental factors, such as temperature, humidity, pulsed magnetic and electric fields, low- and high-frequency electromagnetic fields, light exposure, drop impact, vibration, and splashing. Other characteristics that were tested were alarm volume, clip force, and battery life. The test results were compared with the relevant requirements of three standards: an Ontario Hydro standard for personal alarming dosimeters, an International Electrotechnical Commission draft standard for direct reading personal dose monitors, and an International Electrotechnical Commission standard for thermoluminescence dosimetry systems for personal monitoring. In general, the performance of the Electronic Personal Dosemeter was found to be quite acceptable: it met most of the relevant requirements of the three standards. However, the following deficiencies were found: slow response time; sensitivity to high-frequency electromagnetic fields; poor resistance to dropping; and an alarm that was not loud enough. In addition, the response of the electronic personal dosimeter to low-energy beta rays may be too low for some applications. Problems were experienced with the reliability of operation of the pre-production dosimeters used in these tests. PMID:7790213

  19. Performance and beam characteristics of the Siemens Primus linear accelerator.

    PubMed

    Sontag, M R; Steinberg, T H

    1999-05-01

    Siemens Primus is a small footprint, klystron driven medical linear accelerator incorporating a compact solid state modulator. A double focused multileaf collimator (MLC) replaces the lower jaw. The first Primus in the world was installed at St. Jude Children's Research Hospital in early 1997 with x-ray energies of 6 and 15 MV and electron energies of 8, 10, 12, 15, 18, and 21 MeV. The 10 cm depth dose for a 100 cm SSD 10 X 10 cm2 beam is 68% and 77% for 6 and 15 MV x rays, respectively. For both x-ray energies, beam flatness is slightly better than the manufacturers specification of 3% and beam symmetry is considerably better than 1%. The double focus design of the MLC produces a sharp penumbra (5-7 mm at 6 MV and 6-8 mm at 15 MV), increasing modestly with beam size. MLC leaf leakage is less than 1.25%. The depths of the 80% depth dose for the six electron energies of 8, 10, 12, 15, 18, and 21 MeV are 2.6, 3.2, 4.0, 4.9, 6.0, and 7.4 cm, respectively. Beam flatness is typically 2%-3% for all electron energies except 21 MeV, where it reaches 4% for a 25 X 25 cm2 cone. Electron beam symmetry is better than 1% for all energies except 21 MeV, where it is equal to 1%. The results are stored electronically and may be retrieved using anonymous ftp from the American Institute of Physics, Physics Auxiliary Publication Service. PMID:10360534

  20. 78 FR 19017 - Manufacturer of Controlled Substances; Notice of Registration; Siemens Healthcare Diagnostics, Inc.

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-03-28

    ..., 77 FR 72409, Siemens Healthcare Diagnostics Inc., Attn: RA, 100 GBC Drive, Mail Stop 514, Newark... Tetrahydrocannabinols (7370) I Ecgonine (9180) II Morphine (9300) II The company plans to produce the listed...

  1. 78 FR 41911 - Foreign-Trade Zone 161-Sedgwick County, Kansas; Authorization of Production Activity; Siemens...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-07-12

    ... Activity; Siemens Energy, Inc. (Wind Turbine Nacelles and Hubs); Hutchinson, Kansas On March 7, 2013... (78 FR 20888, April 8, 2013). The FTZ Board has determined that no further review of the activity...

  2. Siemens programmable variable speed DC drives applied to wet and dry expansion engines

    SciTech Connect

    Markley, Daniel J.

    1997-07-01

    This document describes the technical details of the Siemens SIMOREG line of DC variable speed drives as applied to Fermilab wet and dry mechanical expander engines. The expander engines are used throughout the lab in Helium refrigerator installations.

  3. Novel variation and de novo mutation rates in population-wide de novo assembled Danish trios

    PubMed Central

    Besenbacher, Søren; Liu, Siyang; Izarzugaza, José M. G.; Grove, Jakob; Belling, Kirstine; Bork-Jensen, Jette; Huang, Shujia; Als, Thomas D.; Li, Shengting; Yadav, Rachita; Rubio-García, Arcadio; Lescai, Francesco; Demontis, Ditte; Rao, Junhua; Ye, Weijian; Mailund, Thomas; Friborg, Rune M.; Pedersen, Christian N. S.; Xu, Ruiqi; Sun, Jihua; Liu, Hao; Wang, Ou; Cheng, Xiaofang; Flores, David; Rydza, Emil; Rapacki, Kristoffer; Damm Sørensen, John; Chmura, Piotr; Westergaard, David; Dworzynski, Piotr; Sørensen, Thorkild I. A.; Lund, Ole; Hansen, Torben; Xu, Xun; Li, Ning; Bolund, Lars; Pedersen, Oluf; Eiberg, Hans; Krogh, Anders; Børglum, Anders D.; Brunak, Søren; Kristiansen, Karsten; Schierup, Mikkel H.; Wang, Jun; Gupta, Ramneek; Villesen, Palle; Rasmussen, Simon

    2015-01-01

    Building a population-specific catalogue of single nucleotide variants (SNVs), indels and structural variants (SVs) with frequencies, termed a national pan-genome, is critical for further advancing clinical and public health genetics in large cohorts. Here we report a Danish pan-genome obtained from sequencing 10 trios to high depth (50 × ). We report 536k novel SNVs and 283k novel short indels from mapping approaches and develop a population-wide de novo assembly approach to identify 132k novel indels larger than 10 nucleotides with low false discovery rates. We identify a higher proportion of indels and SVs than previous efforts showing the merits of high coverage and de novo assembly approaches. In addition, we use trio information to identify de novo mutations and use a probabilistic method to provide direct estimates of 1.27e−8 and 1.5e−9 per nucleotide per generation for SNVs and indels, respectively. PMID:25597990

  4. Hsc70 chaperone activity underlies Trio GEF function in axon growth and guidance induced by netrin-1

    PubMed Central

    DeGeer, Jonathan; Kaplan, Andrew; Mattar, Pierre; Morabito, Morgane; Stochaj, Ursula; Kennedy, Timothy E.; Debant, Anne; Cayouette, Michel; Fournier, Alyson E.

    2015-01-01

    During development, netrin-1 is both an attractive and repulsive axon guidance cue and mediates its attractive function through the receptor Deleted in Colorectal Cancer (DCC). The activation of Rho guanosine triphosphatases within the extending growth cone facilitates the dynamic reorganization of the cytoskeleton required to drive axon extension. The Rac1 guanine nucleotide exchange factor (GEF) Trio is essential for netrin-1–induced axon outgrowth and guidance. Here, we identify the molecular chaperone heat shock cognate protein 70 (Hsc70) as a novel Trio regulator. Hsc70 dynamically associated with the N-terminal region and Rac1 GEF domain of Trio. Whereas Hsc70 expression supported Trio-dependent Rac1 activation, adenosine triphosphatase–deficient Hsc70 (D10N) abrogated Trio Rac1 GEF activity and netrin-1–induced Rac1 activation. Hsc70 was required for netrin-1–mediated axon growth and attraction in vitro, whereas Hsc70 activity supported callosal projections and radial neuronal migration in the embryonic neocortex. These findings demonstrate that Hsc70 chaperone activity is required for Rac1 activation by Trio and this function underlies netrin-1/DCC-dependent axon outgrowth and guidance. PMID:26323693

  5. Haplotype analysis of the COMT-ARVCF gene region in Israeli anorexia nervosa family trios.

    PubMed

    Michaelovsky, Elena; Frisch, Amos; Leor, Shani; Stein, Daniel; Danziger, Yardena; Carel, Cynthia; Fennig, Silvana; Mimouni, Marc; Klauck, Sabine M; Benner, Axel; Poustka, Annemarie; Apter, Alan; Weizman, Abraham

    2005-11-01

    Anorexia nervosa (AN) is a severe and complex psychiatric disorder with a significant genetic contribution. Previously, we found an association between AN and the 158Val/Met polymorphism of the catechol-O-methyltransferase (COMT) gene in a family-based study of 51 Israeli AN trios. In the present study, we extended the original sample to include 85 family trios [66 AN restricting (AN-R) and 19 bingeing/purging (AN-BP) subtype] and performed a family-based transmission disequilibrium test (TDT) analysis for five SNPs in the COMT and two in the adjacent ARVCF gene. Association was found between AN-R and several SNPs in the COMT-ARVCF region including the 158Val/Met polymorphism. TDT analysis of 5-SNP haplotypes in AN-R trios revealed an overall statistically significant transmission disequilibrium (P < 0.001). Specifically, haplotype B [COMT-186C-408G-472G(158Val)-ARVCF-659C(220Pro)-524T(175Val)] was preferentially transmitted (P < 0.001) from parents of AN-R patients to their affected daughters, while haplotype A [COMT-186T-408C-472A(158Met)-ARVCF-659T(220Leu)-524C(175Ala)] was preferentially (P = 0.01) not transmitted. Haplotype B was associated with increased risk (RR 3.38; 0.95CI 1.98-6.43) while haplotype A exhibited a protective effect (RR 0.40; 0.95CI 0.21-0.70) for AN-R. Preferential transmission of the risk alleles and haplotypes from the parents was mostly contributed by the fathers. No significant transmission disequilibrium of alleles or haplotypes was found for AN-BP trios. The risk and protective haplotypes may carry molecular variations in the COMT gene or its vicinity that are relevant to the pathophysiology of restrictive anorexia nervosa in the Israeli-Jewish population. PMID:16118784

  6. The bacterial DnaA-trio replication origin element specifies single-stranded DNA initiator binding.

    PubMed

    Richardson, Tomas T; Harran, Omar; Murray, Heath

    2016-06-16

    DNA replication is tightly controlled to ensure accurate inheritance of genetic information. In all organisms, initiator proteins possessing AAA+ (ATPases associated with various cellular activities) domains bind replication origins to license new rounds of DNA synthesis. In bacteria the master initiator protein, DnaA, is highly conserved and has two crucial DNA binding activities. DnaA monomers recognize the replication origin (oriC) by binding double-stranded DNA sequences (DnaA-boxes); subsequently, DnaA filaments assemble and promote duplex unwinding by engaging and stretching a single DNA strand. While the specificity for duplex DnaA-boxes by DnaA has been appreciated for over 30 years, the sequence specificity for single-strand DNA binding has remained unknown. Here we identify a new indispensable bacterial replication origin element composed of a repeating trinucleotide motif that we term the DnaA-trio. We show that the function of the DnaA-trio is to stabilize DnaA filaments on a single DNA strand, thus providing essential precision to this binding mechanism. Bioinformatic analysis detects DnaA-trios in replication origins throughout the bacterial kingdom, indicating that this element is part of the core oriC structure. The discovery and characterization of the novel DnaA-trio extends our fundamental understanding of bacterial DNA replication initiation, and because of the conserved structure of AAA+ initiator proteins these findings raise the possibility of specific recognition motifs within replication origins of higher organisms. PMID:27281207

  7. Design, fabrication and testing of the gas analysis system for the tritium recovery experiment, TRIO-01

    SciTech Connect

    Finn, P.A.; Reedy, G.T.; Homa, M.I.; Clemmer, R.G.; Pappas, G.; Slawecki, M.A.; Graczyk, D.G.; Bowers, D.L.; Clemmer, E.D.

    1983-01-01

    The tritium recovery experiment, TRIO-01, required a gas analysis system which detected the form of tritium, the amount of tritium (differential and integral), and the presence and amount of other radioactive species. The system had to handle all contingencies and function for months at a time unattended during weekend operation. The designed system, described herein, consisted of a train of components which could be grouped as desired to match tritium release behavior.

  8. A gradient-boosting approach for filtering de novo mutations in parent–offspring trios

    PubMed Central

    Liu, Yongzhuang; Li, Bingshan; Tan, Renjie; Zhu, Xiaolin; Wang, Yadong

    2014-01-01

    Motivation: Whole-genome and -exome sequencing on parent–offspring trios is a powerful approach to identifying disease-associated genes by detecting de novo mutations in patients. Accurate detection of de novo mutations from sequencing data is a critical step in trio-based genetic studies. Existing bioinformatic approaches usually yield high error rates due to sequencing artifacts and alignment issues, which may either miss true de novo mutations or call too many false ones, making downstream validation and analysis difficult. In particular, current approaches have much worse specificity than sensitivity, and developing effective filters to discriminate genuine from spurious de novo mutations remains an unsolved challenge. Results: In this article, we curated 59 sequence features in whole genome and exome alignment context which are considered to be relevant to discriminating true de novo mutations from artifacts, and then employed a machine-learning approach to classify candidates as true or false de novo mutations. Specifically, we built a classifier, named De Novo Mutation Filter (DNMFilter), using gradient boosting as the classification algorithm. We built the training set using experimentally validated true and false de novo mutations as well as collected false de novo mutations from an in-house large-scale exome-sequencing project. We evaluated DNMFilter’s theoretical performance and investigated relative importance of different sequence features on the classification accuracy. Finally, we applied DNMFilter on our in-house whole exome trios and one CEU trio from the 1000 Genomes Project and found that DNMFilter could be coupled with commonly used de novo mutation detection approaches as an effective filtering approach to significantly reduce false discovery rate without sacrificing sensitivity. Availability: The software DNMFilter implemented using a combination of Java and R is freely available from the website at http

  9. An Efficient Trio-Based Mini-Haplotyping Method for Genetic Diagnosis of Phenylketonuria

    PubMed Central

    Talebi, Saeed; Entezam, Mona; Mohajer, Neda; Kazemi-sefat, Golnaz-Ensieh; Razipour, Masoumeh; Ahmadloo, Somayeh; Setoodeh, Aria; Keramatipour, Mohammad

    2016-01-01

    Objective The phenylalanine hydroxylase (PAH) locus has high linkage disequilibrium. Haplotypes related to this locus may thus be considered sufficiently informative for genetic diagnosis and carrier screening using multi-allelic markers. In this study, we present an efficient method for haplotype analysis of PAH locus using multiplexing dyes. In addition, we explain how to resolve the dye shift challenge in multiplex short tandem repeat (STR) genotyping. Materials and Methods One hundred family trios were included in this descriptive study. The forward primer of a tetra-nucleotide STR and the reverse primer of a variable number tandem repeat (VNTR) were labeled with three different non-overlapping dyes 5-carboxyfluorescein (FAM), 6-carboxy-N,N,N’,N’-tetramethylrhodamine (HEX) and 6-carboxy-N,N,N’,N’-tetramethylrhodamine (TAMRA). The polymerase chain reaction (PCR) products from each family trio were multiplexed for capillary electrophoresis and results were analyzed using Peak Scanner software. Results Multiplexing trio products decreased the cost significantly. The TAMRA labeled products had a significant predictable shift (migrated at a slower electrophoretic rate) relative to the HEX and FAM labeled products. Through our methodology we achieve, the less inter-dye shift than intra-dye shift variance. Correcting the dye shift in the labeled products, according to the reference allele size, significantly decreased the inter-dye variability (P<0.001). Conclusion Multiplexing trio products helps to detect and resolve the dye shift accurately in each family, which otherwise would result in diagnostic error. The dye system of FAM, HEX and TAMRA is more feasible and cheaper than other dye systems. PMID:27540528

  10. Assessing and comparison of different machine learning methods in parent-offspring trios for genotype imputation.

    PubMed

    Mikhchi, Abbas; Honarvar, Mahmood; Kashan, Nasser Emam Jomeh; Aminafshar, Mehdi

    2016-06-21

    Genotype imputation is an important tool for prediction of unknown genotypes for both unrelated individuals and parent-offspring trios. Several imputation methods are available and can either employ universal machine learning methods, or deploy algorithms dedicated to infer missing genotypes. In this research the performance of eight machine learning methods: Support Vector Machine, K-Nearest Neighbors, Extreme Learning Machine, Radial Basis Function, Random Forest, AdaBoost, LogitBoost, and TotalBoost compared in terms of the imputation accuracy, computation time and the factors affecting imputation accuracy. The methods employed using real and simulated datasets to impute the un-typed SNPs in parent-offspring trios. The tested methods show that imputation of parent-offspring trios can be accurate. The Random Forest and Support Vector Machine were more accurate than the other machine learning methods. The TotalBoost performed slightly worse than the other methods.The running times were different between methods. The ELM was always most fast algorithm. In case of increasing the sample size, the RBF requires long imputation time.The tested methods in this research can be an alternative for imputation of un-typed SNPs in low missing rate of data. However, it is recommended that other machine learning methods to be used for imputation. PMID:27049046

  11. Results from Operational Testing of the Siemens Smart Grid-Capable Electric Vehicle Supply Equipment

    SciTech Connect

    Bennett, Brion

    2015-05-01

    The Idaho National Laboratory conducted testing and analysis of the Siemens smart grid capable electric vehicle supply equipment (EVSE), which was a deliverable from Siemens for the U.S. Department of Energy FOA-554. The Idaho National Laboratory has extensive knowledge and experience in testing advanced conductive and wireless charging systems though INL’s support of the U.S. Department of Energy’s Advanced Vehicle Testing Activity. This document details the findings from the EVSE operational testing conducted at the Idaho National Laboratory on the Siemens smart grid capable EVSE. The testing conducted on the EVSE included energy efficiency testing, SAE J1772 functionality testing, abnormal conditions testing, and charging of a plug-in vehicle.

  12. Normal findings on brain FLAIR MRI scans at 3T

    PubMed Central

    Neema, Mohit; Guss, Zachary D.; Stankiewicz, James M.; Arora, Ashish; Healy, Brian C.; Bakshi, Rohit

    2010-01-01

    BACKGROUND AND PURPOSE Fluid attenuated inversion recovery (FLAIR) MR imaging of the brain has become a routine tool for assessing lesions in patients with suspected neurologic disorders. There is growing interest in 3T brain FLAIR MR imaging but little normative data are available. The purpose of this study was to evaluate the frequency and topography of cerebral hyperintensities seen with FLAIR MR imaging of the brain at 3T in a normal population and compare those findings to 1.5T. MATERIALS AND METHODS Whole-brain 2D FLAIR MR imaging was performed in 22 healthy controls (mean age, 44 ± 8 years; range, 30–53 years) at 3T. Fifteen of these subjects also underwent 2D FLAIR at 1.5T, with similar optimized parameters and voxel size. Cerebral hyperintense areas, including discrete foci, anterior and posterior periventricular capping, diffuse parenchymal hyperintensity, septal hyperintensity, corticospinal tract hyperintensity, and CSF flow artifacts were assessed. The Spearman rank test assessed the correlation between discrete hyperintense foci and age. The Wilcoxon signed rank test compared foci detectability at 3T versus 1.5T. RESULTS FLAIR at 3T commonly showed hyperintensities such as discrete foci (mean, 10.68 per subject; at least 1 present in 68% of subjects), anterior and posterior periventricular capping, diffuse posterior white matter hyperintensity, septal hyperintensity, corticospinal tract hyperintensity, and ventricular CSF flow artifacts. FLAIR at 3T showed a higher hyperintense foci volume (170 ± 243 versus 93 ± 152 mm3, P < .01) and number (9.4 ± 13 versus 5.5 ± 9.2, P < .01) than at 1.5T. No significant differences (P = .68) in the length/diameter of individual discrete hyperintense foci were seen between 3T and 1.5T. Discrete foci volume (r = 0.72 at 3T, r = 0.70 at 1.5T) and number (r = 0.74 at 3T; r = 0.69 at 1.5T) correlated with age to a similar degree on both platforms. All discrete foci were confined to the noncallosal supratentorial

  13. Clinical 3T MR imaging: mastering the challenges.

    PubMed

    Tanenbaum, Lawrence N

    2006-02-01

    3T MRI is ready to meet the needs of clinical practice. SAR limitations are minimized by technical advances and surface coils are available for all core applications. With appropriate adjustments to scanning protocols, one can master the challenges of scanning at 3T; studies of the brain, spine, chest, abdomen, pelvis, vasculature, and extremities can be consistently higher in quality than are those obtained at 1.5T. The superior studies that are obtainable at 3T have great appeal to clinicians who are sophisticated about MR technology in areas, such as neurology, orthopedics, vascular surgery, and oncology,and encourage a shift in referrals toward practices that invest in higher field technology. The greater sensitivity to magnetic susceptibility offers unique benefits in functional neuroimaging, and available software/hardware packages enhance clinical setting feasibility, which adds a source of new referrals. The greater overall signal of 3T can be manipulated to make scanning more comfortable and with less motion artifact because scan times could be half as long. Spectacular anatomic delineation that is provided by high-definition scanning at true 1024 resolution can improve preoperative assessment and may improve sensitivity to smaller lesions. 3T provides practices with an advantage that is sought increasingly by high field strength purchasers in a competitive market. Only cost considerations stand in the way of the eventual domination of 3T systems in the high field strength market. PMID:16530631

  14. A Trio of modulators for the Fermilab tevatron electron lens project

    SciTech Connect

    David W. Wildman et al.

    2001-07-25

    Three high voltage modulators used during testing and operation of the Tevatron Electron Lens (TEL) at Fermilab will be described. Short high voltage (0 to {approximately} 20kV) pulses from these modulators vary the anode-cathode voltage of the TEL electron gun to control the magnitude of the electron beam current. The trio of modulators include a low repetition rate MOSFET-based pulser, a fast ionization device, and a high average power tetrode modulator. The characteristics of each device will be discussed and typical outputs from each type of modulator is shown.

  15. Power of sib-pair and sib-trio linkage analysis with assortative mating and multiple disease loci

    SciTech Connect

    Sribney, W.M.; Swift, M. )

    1992-10-01

    Sib-pair linkage analysis has been proposed for identifying genes that predispose to common diseases. The authors have shown that the presence of assortative mating and multiple disease-susceptibility loci (genetic heterogeneity) can increase the required sample size for affected-affected sib pairs several fold over the sample size required under random mating. They propose a new test statistic based on sib trios composed of either one unaffected and two affected siblings or one affected and two unaffected siblings. The sample-size requirements under assortative mating and multiple disease loci for these sib-trio statistics are much smaller, under most conditions, than the corresponding sample sizes for sib pairs. Study designs based on data from sib trios with one or two affected members are recommended whenever assortative mating and genetic heterogeneity are suspected. 31 refs.

  16. ETV Report:Siemens Model H-4XE-HO Open Channel UV System

    EPA Science Inventory

    Verification testing of the Siemens Barrier Sunligt H-4XE-HO UV System was completed at the UV Validation and Research Center of New York (UV Center), located in Johnstown, NY. The H-4XE System utilizes 16 high-output, low-pressure lamps oriented horizontally and parallel to the...

  17. Basic dosimetric verification in water of the anisotropic analytical algorithm for Varian, Elekta and Siemens linacs.

    PubMed

    Cozzi, Luca; Nicolini, Giorgia; Vanetti, Eugenio; Clivio, Alessandro; Glashörster, Marco; Schiefer, Hans; Fogliata, Antonella

    2008-01-01

    Since early 2007 a new version of the Anisotropic Analytical Algorithm (AAA) for photon dose calculations was released by Varian Medical Systems for clinical usage on Elekta linacs and also, with some restrictions, for Siemens linacs. Basic validation studies were performed and reported for three beams. 4,6 and 15 MV for an Elekta Synergy, 6 and 15 MV for a Siemens Primus and, as a reference, for 6 and 15 MV from a Varian Clinac 2100C/D. Generally AAA calculations reproduced well measured data and small deviations were observed for open and wedged fields. PDD curves showed in average differences between calculation and measurement smaller than 1% or 1.2 mm for Elekta beams, 1% or 1.8 mm for Siemens beams and 1% or 1 mm for Varian beams. Profiles in the flattened region matched measurements with deviations smaller than 1% for Elekta and Varian beams, 2% for Siemens. Percentage differences in Output Factors were observed as small as 1% in average. PMID:18705613

  18. An Allele of Sequoia Dominantly Enhances a Trio Mutant Phenotype to Influence Drosophila Larval Behavior

    PubMed Central

    Liebl, Eric C.

    2013-01-01

    The transition of Drosophila third instar larvae from feeding, photo-phobic foragers to non-feeding, photo-neutral wanderers is a classic behavioral switch that precedes pupariation. The neuronal network responsible for this behavior has recently begun to be defined. Previous genetic analyses have identified signaling components for food and light sensory inputs and neuropeptide hormonal outputs as being critical for the forager to wanderer transition. Trio is a Rho-Guanine Nucleotide Exchange Factor integrated into a variety of signaling networks including those governing axon pathfinding in early development. Sequoia is a pan-neuronally expressed zinc-finger transcription factor that governs dendrite and axon outgrowth. Using pre-pupal lethality as an endpoint, we have screened for dominant second-site enhancers of a weakly lethal trio mutant background. In these screens, an allele of sequoia has been identified. While these mutants have no obvious disruption of embryonic central nervous system architecture and survive to third instar larvae similar to controls, they retain forager behavior and thus fail to pupariate at high frequency. PMID:24376789

  19. Quantifiler® Trio Kit and forensic samples management: a matter of degradation.

    PubMed

    Vernarecci, Stefano; Ottaviani, Enrica; Agostino, Alessandro; Mei, Elisabetta; Calandro, Lisa; Montagna, Paola

    2015-05-01

    DNA collected from crime scenes may have experienced different levels of degradation. This is mainly due to sample exposure to different environmental factors. The impact of DNA degradation on short tandem repeat (STR) profiling can lead to partial or null information and in some cases, the identification of the trace may fail. The availability of a system enabling the assessment not only of the quantity of the DNA but also of its quality in terms of degradation would result in shorter time for sample processing, more reliable identifications and cost reduction by predicting the quality of the DNA profiles prior to STR analysis. We report here a study on 181 selected degraded DNA samples extracted from real crime scene evidence. The selected samples were processed by combining the use of a new commercial quantification kit (Quantifiler® Trio) with a new 24 marker multiplex PCR amplification kit (Globalfiler® Kit). Applying different statistical analyses we investigated the reliability of the Degradation Index provided by the Quantifiler® Trio in determining the level of DNA degradation in a forensic sample. This useful information can be used to predict the quality of the profile obtained after STR amplification. The combination of such a quantification kit with different PCR protocols allowed us to define practical guidelines for processing degraded forensic DNA samples with a simplified and comprehensive approach. PMID:25544252

  20. Teaching Trio

    ERIC Educational Resources Information Center

    Olson, Cathy Applefeld

    2012-01-01

    For the Tower-Tannerts of Texas, music education runs in the family, and that's been the case for more than 100 years. Matriarch Mollie Gregory Tower is a lecturer at Texas State University and a former elementary school teacher, textbook writer, music supervisor, and president of the Texas Music Educators Conference (TMEC). Her daughter Debbie…

  1. Conditioning adaptive combination of P-values method to analyze case-parent trios with or without population controls

    PubMed Central

    Lin, Wan-Yu; Liang, Yun-Chieh

    2016-01-01

    Detection of rare causal variants can help uncover the etiology of complex diseases. Recruiting case-parent trios is a popular study design in family-based studies. If researchers can obtain data from population controls, utilizing them in trio analyses can improve the power of methods. The transmission disequilibrium test (TDT) is a well-known method to analyze case-parent trio data. It has been extended to rare-variant association testing (abbreviated as “rvTDT”), with the flexibility to incorporate population controls. The rvTDT method is robust to population stratification. However, power loss may occur in the conditioning process. Here we propose a “conditioning adaptive combination of P-values method” (abbreviated as “conADA”), to analyze trios with/without unrelated controls. By first truncating the variants with larger P-values, we decrease the vulnerability of conADA to the inclusion of neutral variants. Moreover, because the test statistic is developed by conditioning on parental genotypes, conADA generates valid statistical inference in the presence of population stratification. With regard to statistical methods for next-generation sequencing data analyses, validity may be hampered by population stratification, whereas power may be affected by the inclusion of neutral variants. We recommend conADA for its robustness to these two factors (population stratification and the inclusion of neutral variants). PMID:27341039

  2. Composite fermions, trios, and quartets in a Fermi-Bose mixture

    SciTech Connect

    Kagan, M.Yu.; Brodsky, I.V.; Efremov, D.V.; Klaptsov, A.V.

    2004-08-01

    We consider a model of a Fermi-Bose mixture with strong hard-core repulsion between particles of the same sort and attraction between particles of different sorts. In this case, besides the standard anomalous averages of the type , , and , a pairing between fermions and bosons of the type bc is possible. This pairing corresponds to the creation of composite fermions in the system. At low temperatures and equal densities of fermions and bosons composite fermions are further paired in quartets. At higher temperatures trios, which consist of composite fermions and elementary bosons, are also present in the system. Our investigations are important in connection with the recent observation of weakly bound dimers in magnetic and optical dipole traps at ultralow temperatures and with the observation of the collapse of a Fermi gas in an attractive Fermi-Bose mixture of neutral particles.

  3. [Frequency quotient of paternity index Qf in a case of replacement of the natural father within a trio with his brother].

    PubMed

    Berent, Jarosław; Jacewicz, Renata; Szram, Stefan

    2005-01-01

    Cases of trios in which the natural father's near blood-relative instead of the natural father are among the hardest to give an expert opinion on. The goal of this study was to determine with what frequencies specific given values of paternity index appear in trios with the natural father and with the natural father's brother, and whether these frequencies can be the basis for discovering the substitution of the natural father by a brother in a given trio examined. The study material was the population of genotypes investigated in the AmpF/STR SGM Plus system. It was seen that the higher the paternity index values, the more frequently they appear in trios with the natural father, and less frequently in trios with the natural father's brother. This dependency has a general character and obtained in all the systems studied. The frequency quotients Qf (defined as the frequency of a given PI value in trios with the natural father in relation to the "reduced" frequency of the PI value in trios with the brother of the natural father) are found in the region of c. 0.6 to c. 1.4, whereby extreme values are rare, and the most frequent are cases where Qf oscillates within modest limits around the value 1. This fact shows that the majority of cases, calculated Qf values contribute little to resolving the question of whether the defendant--or his brother--is the child's father. PMID:16080432

  4. Sinusoidal Siemens star spatial frequency response measurement errors due to misidentified target centers

    SciTech Connect

    Birch, Gabriel Carisle; Griffin, John Clark

    2015-07-23

    Numerous methods are available to measure the spatial frequency response (SFR) of an optical system. A recent change to the ISO 12233 photography resolution standard includes a sinusoidal Siemens star test target. We take the sinusoidal Siemens star proposed by the ISO 12233 standard, measure system SFR, and perform an analysis of errors induced by incorrectly identifying the center of a test target. We show a closed-form solution for the radial profile intensity measurement given an incorrectly determined center and describe how this error reduces the measured SFR of the system. As a result, using the closed-form solution, we propose a two-step process by which test target centers are corrected and the measured SFR is restored to the nominal, correctly centered values.

  5. Sinusoidal Siemens star spatial frequency response measurement errors due to misidentified target centers

    DOE PAGESBeta

    Birch, Gabriel Carisle; Griffin, John Clark

    2015-07-23

    Numerous methods are available to measure the spatial frequency response (SFR) of an optical system. A recent change to the ISO 12233 photography resolution standard includes a sinusoidal Siemens star test target. We take the sinusoidal Siemens star proposed by the ISO 12233 standard, measure system SFR, and perform an analysis of errors induced by incorrectly identifying the center of a test target. We show a closed-form solution for the radial profile intensity measurement given an incorrectly determined center and describe how this error reduces the measured SFR of the system. As a result, using the closed-form solution, we proposemore » a two-step process by which test target centers are corrected and the measured SFR is restored to the nominal, correctly centered values.« less

  6. GeoEtrim, SharpQ and Epix: Trio of Tools for Geospatial Image Analysis

    NASA Astrophysics Data System (ADS)

    Topan, H.; Özendi, M.; Cam, A.; Oruç, M.

    2014-09-01

    This paper presents an overview of three approaches developed in Matlab for geospatial analysis of images. GeoEtrim has two subpackages, GeoSpot and GeoFigcon. GeoSpot aims to perform bundle adjustment of stereo linear array remotely sensed images considering their interior and exterior orientation parameters. The correlation among orientation parameters, their validation and efficiency on the final accuracy can be estimated. The current version is available for SPOT-5 HRG level 1A stereoimages, reaching ±1 pixel accuracy at ICPs (Independent Check Points). GeoFigcon is the other sub-package of GeoEtrim, developed for estimation of georeferencing accuracy of orthoimages generated by various sensor-independent mathematical models and RFM (Rational Function Model). Using GeoFigcon, one can estimate the combined effect of the accuracy of transformation parameters estimated/updated by GCPs and DEM accuracy on the georeferencing accuracy of orthoimages. The experiments with IKONOS Geo, QuickBird OrthoReady Standard, OrbView-3 Basic and Pléiades-1A Primary prove that using RFM produces higher accuracy than using sensor-independent transformation models. Moreover, accuracy varies reducing from geometric centre of GCPs accommodating the high profile of topography. SharpQ generates pan-sharp images using the methods PCA (Principal Component Analysis), Brovey, and IHS (Intensity Hue Saturation), validates their quality with quantitative analysis by the methods CC (Correlation Coefficient), RMSE (Root Mean Square Error), RASE (Relative Average Spectral Error), SAM (Spectral Angle Mapper) and ERGAS (Erreur Relative Globale Adimensionnelle de Synthése). epix, the last member of the trio, can be used for estimation of effective GSD (Ground Sampling Distance) value of original or generated (such as pan-sharp) images, depending the principle of ESF (Edge Spread Function). So the real geometric resolution can be estimated for any kind of image. This trio is being still developed

  7. TRIO Platform: A Novel Low Profile In vivo Imaging Support and Restraint System for Mice

    PubMed Central

    Voziyanov, Vladislav; Kemp, Benjamin S.; Dressel, Chelsea A.; Ponder, Kayla; Murray, Teresa A.

    2016-01-01

    High resolution, in vivo optical imaging of the mouse brain over time often requires anesthesia, which necessitates maintaining the animal's body temperature and level of anesthesia, as well as securing the head in an optimal, stable position. Controlling each parameter usually requires using multiple systems. Assembling multiple components into the small space on a standard microscope stage can be difficult and some commercially available parts simply do not fit. Furthermore, it is time-consuming to position an animal in the identical position over multiple imaging sessions for longitudinal studies. This is especially true when using an implanted gradient index (GRIN) lens for deep brain imaging. The multiphoton laser beam must be parallel with the shaft of the lens because even a slight tilt of the lens can degrade image quality. In response to these challenges, we have designed a compact, integrated in vivo imaging support system to overcome the problems created by using separate systems during optical imaging in mice. It is a single platform that provides (1) sturdy head fixation, (2) an integrated gas anesthesia mask, and (3) safe warm water heating. This THREE-IN-ONE (TRIO) Platform has a small footprint and a low profile that positions a mouse's head only 20 mm above the microscope stage. This height is about one half to one third the height of most commercially available immobilization devices. We have successfully employed this system, using isoflurane in over 40 imaging sessions with an average of 2 h per session with no leaks or other malfunctions. Due to its smaller size, the TRIO Platform can be used with a wider range of upright microscopes and stages. Most of the components were designed in SOLIDWORKS® and fabricated using a 3D printer. This additive manufacturing approach also readily permits size modifications for creating systems for other small animals. PMID:27199633

  8. Paternity investigation among known false trios: ABO, Rh, MNSs, Kell, Duffy, Kidd, and HLA systems.

    PubMed

    Salaru, N N

    1993-11-01

    This paternity study was performed with trios in which the putative father was not the biological father (NBF), in order to evaluate adjustment of genetic markers employed to disclose non biological fathers for the population, and the biological meaning of likelihood of paternity in casework. All 923 generated trios had ABO, Rh, MNS, Kell and HLA systems tested; 372 of them also had Duffy and Kidd systems tested. The most powerful exclusion system was HLA, followed, in this order, by ABO, Rh, Duffy, MNSs, Kidd, and Kell. Taking into account the Indian/black/white historical miscegenation background in the population, an improvement in the performance of red blood cells as disclosers of non biological fathers could be achieved, if particular additional sera were used. In the group tested with seven different systems, direct exclusions were observed in 90.31%, and they were single system exclusions in 26.61%. In order to avoid the remote possibility of mutation, it is suggested that the number of used systems be increased. Indirect exclusions were verified in 8.87% and only 0.81% of NBF were not excluded at all. In this last group, probabilities of paternity were calculated and two values greater than 95% were obtained. To be able to accomplish the "visum et repertum" duty and to assist the court, the expert should equally emphasize: a) the probability of paternity of the alleged father and the possibility of finding an unexcluded NBF; b) the actual performance of systems used to uncover NBF, together with the probabilities of paternity of those who were not discovered; c) the previous referenced trend of probabilities of paternity of true and of non-biological fathers to cluster in distinct class intervals of likelihood of paternity. PMID:8263491

  9. TRIO Platform: A Novel Low Profile In vivo Imaging Support and Restraint System for Mice.

    PubMed

    Voziyanov, Vladislav; Kemp, Benjamin S; Dressel, Chelsea A; Ponder, Kayla; Murray, Teresa A

    2016-01-01

    High resolution, in vivo optical imaging of the mouse brain over time often requires anesthesia, which necessitates maintaining the animal's body temperature and level of anesthesia, as well as securing the head in an optimal, stable position. Controlling each parameter usually requires using multiple systems. Assembling multiple components into the small space on a standard microscope stage can be difficult and some commercially available parts simply do not fit. Furthermore, it is time-consuming to position an animal in the identical position over multiple imaging sessions for longitudinal studies. This is especially true when using an implanted gradient index (GRIN) lens for deep brain imaging. The multiphoton laser beam must be parallel with the shaft of the lens because even a slight tilt of the lens can degrade image quality. In response to these challenges, we have designed a compact, integrated in vivo imaging support system to overcome the problems created by using separate systems during optical imaging in mice. It is a single platform that provides (1) sturdy head fixation, (2) an integrated gas anesthesia mask, and (3) safe warm water heating. This THREE-IN-ONE (TRIO) Platform has a small footprint and a low profile that positions a mouse's head only 20 mm above the microscope stage. This height is about one half to one third the height of most commercially available immobilization devices. We have successfully employed this system, using isoflurane in over 40 imaging sessions with an average of 2 h per session with no leaks or other malfunctions. Due to its smaller size, the TRIO Platform can be used with a wider range of upright microscopes and stages. Most of the components were designed in SOLIDWORKS® and fabricated using a 3D printer. This additive manufacturing approach also readily permits size modifications for creating systems for other small animals. PMID:27199633

  10. Pinnacle Health / Zynx Health / Siemens Medical Solutions A Study of Integration of Evidence Based Nursing Content

    PubMed Central

    Matter, Sheri; Brown, Cindy; Button, Patricia S.; Kennedy, Rosemary

    2006-01-01

    In 2005, Pinnacle Health System, Zynx Health, and Siemens Medical Solutions developed a partnership to conduct a study to explore the opportunities and challenges associated with the integration of evidence-based knowledge within the EHR with the goal of creating repeatable methodologies for integrating nursing knowledge within the EHR. The two-phase study involved access to referential evidence-based content, as well as integration of customized evidence-based plans of care within the documentation applications of the EHR.

  11. Plasma system of the GOL-3T facility

    SciTech Connect

    Arzhannikov, A. V.; Burdakov, A. V.; Burmasov, V. S.; Ivanov, I. A.; Kuznetsov, S. A.; Kuklin, K. N.; Mekler, K. I.; Polosatkin, S. V.; Postupaev, V. V. Rovenskikh, A. F.; Sinitsky, S. L.; Sklyarov, V. F.

    2015-11-15

    The plasma system and diagnostics of the new facility GOL-3T are described. This facility is the final result of the first stage in the deep upgrade of the GOL-3 multiple-mirror system, which has operated at the Budker Institute of Nuclear Physics since 1988. The upgrade project supposes creation of two new independent facilities at the site of GOL-3. The GOL-3T facility is intended to study the physics of beam—plasma interaction and generation of subterahertz electromagnetic radiation during the collective relaxation of a high-power relativistic electron beam with a duration of 5–10 μs. Studies on the physics of multiple-mirror plasma confinement in axisymmetric magnetic systems will be continued in a new range of experiment parameters at the second facility, named GOL-NB.

  12. SIEMENS ADVANCED QUANTRA FTICR MASS SPECTROMETER FOR ULTRA HIGH RESOLUTION AT LOW MASS

    SciTech Connect

    Spencer, W; Laura Tovo, L

    2008-07-08

    The Siemens Advanced Quantra Fourier Transform Ion Cyclotron Resonance (FTICR) mass spectrometer was evaluated as an alternative instrument to large double focusing mass spectrometers for gas analysis. High resolution mass spectrometers capable of resolving the common mass isomers of the hydrogen isotopes are used to provide data for accurate loading of reservoirs and to monitor separation of tritium, deuterium, and helium. Conventional double focusing magnetic sector instruments have a resolution that is limited to about 5000. The Siemens FTICR instrument achieves resolution beyond 400,000 and could possibly resolve the tritium ion from the helium-3 ion, which differ by the weight of an electron, 0.00549 amu. Working with Y-12 and LANL, SRNL requested Siemens to modify their commercial Quantra system for low mass analysis. To achieve the required performance, Siemens had to increase the available waveform operating frequency from 5 MHz to 40 MHz and completely redesign the control electronics and software. However, they were able to use the previous ion trap, magnet, passive pump, and piezo-electric pulsed inlet valve design. NNSA invested $1M in this project and acquired four systems, two for Y-12 and one each for SRNL and LANL. Siemens claimed a $10M investment in the Quantra systems. The new Siemens Advanced Quantra demonstrated phenomenal resolution in the low mass range. Resolution greater than 400,000 was achieved for mass 2. The new spectrometer had a useful working mass range to 500 Daltons. However, experiments found that a continuous single scan from low mass to high was not possible. Two useful working ranges were established covering masses 1 to 6 and masses 12 to 500 for our studies. A compromise performance condition enabled masses 1 to 45 to be surveyed. The instrument was found to have a dynamic range of about three orders of magnitude and quantitative analysis is expected to be limited to around 5 percent without using complex fitting algorithms

  13. Rare erroneous results on the Siemens Dimension Vista® platform due to urine carryover: A warning to current users.

    PubMed

    Karger, Amy B; Senn, Christine; Skogseth, Kathy; Floodman, Stacy

    2016-07-01

    In 2014, the West Bank Laboratory at the University of Minnesota, part of the non-profit Fairview Health Services system, went live with two Siemens Dimension Vista® 500 instruments. In the first few months after go-live, the lab began receiving reports of rare, erroneous results from clinicians. After further investigation it was noted that most of the errors being reported followed a consistent pattern with significantly elevated potassium, BUN and creatinine. After months of unsuccessful troubleshooting with Siemens, our lab and others within the Fairview Health Services system began hypothesizing that urine carryover was occurring due to the pattern of elevated analytes, and asked Siemens to investigate this hypothesis. In March 2016 Siemens confirmed a software defect which omits an aliquot probe rinse resulting in sample carryover in rare cases. Our objective is to alert current users of the Siemens Dimension Vista® instruments to this rare but alarming phenomenon, discuss the frequency and impact of the erroneous results at our institution, and detail recommended steps for users of Siemens Dimension Vista® instruments to help retrospectively determine whether they have experienced similar erroneous results. PMID:27174361

  14. Standardized quantitative measurements of wrist cartilage in healthy humans using 3T magnetic resonance imaging

    PubMed Central

    Zink, Jean-Vincent; Souteyrand, Philippe; Guis, Sandrine; Chagnaud, Christophe; Fur, Yann Le; Militianu, Daniela; Mattei, Jean-Pierre; Rozenbaum, Michael; Rosner, Itzhak; Guye, Maxime; Bernard, Monique; Bendahan, David

    2015-01-01

    AIM: To quantify the wrist cartilage cross-sectional area in humans from a 3D magnetic resonance imaging (MRI) dataset and to assess the corresponding reproducibility. METHODS: The study was conducted in 14 healthy volunteers (6 females and 8 males) between 30 and 58 years old and devoid of articular pain. Subjects were asked to lie down in the supine position with the right hand positioned above the pelvic region on top of a home-built rigid platform attached to the scanner bed. The wrist was wrapped with a flexible surface coil. MRI investigations were performed at 3T (Verio-Siemens) using volume interpolated breath hold examination (VIBE) and dual echo steady state (DESS) MRI sequences. Cartilage cross sectional area (CSA) was measured on a slice of interest selected from a 3D dataset of the entire carpus and metacarpal-phalangeal areas on the basis of anatomical criteria using conventional image processing radiology software. Cartilage cross-sectional areas between opposite bones in the carpal region were manually selected and quantified using a thresholding method. RESULTS: Cartilage CSA measurements performed on a selected predefined slice were 292.4 ± 39 mm2 using the VIBE sequence and slightly lower, 270.4 ± 50.6 mm2, with the DESS sequence. The inter (14.1%) and intra (2.4%) subject variability was similar for both MRI methods. The coefficients of variation computed for the repeated measurements were also comparable for the VIBE (2.4%) and the DESS (4.8%) sequences. The carpus length averaged over the group was 37.5 ± 2.8 mm with a 7.45% between-subjects coefficient of variation. Of note, wrist cartilage CSA measured with either the VIBE or the DESS sequences was linearly related to the carpal bone length. The variability between subjects was significantly reduced to 8.4% when the CSA was normalized with respect to the carpal bone length. CONCLUSION: The ratio between wrist cartilage CSA and carpal bone length is a highly reproducible standardized

  15. 3-T MR-guided brachytherapy for gynecologic malignancies.

    PubMed

    Kapur, Tina; Egger, Jan; Damato, Antonio; Schmidt, Ehud J; Viswanathan, Akila N

    2012-11-01

    Gynecologic malignancies are a leading cause of death in women worldwide. Standard treatment for many primary and recurrent gynecologic cancer cases includes external-beam radiation followed by brachytherapy. Magnetic resonance (MR) imaging is beneficial in diagnostic evaluation, in mapping the tumor location to tailor radiation dose and in monitoring the tumor response to treatment. Initial studies of MR guidance in gynecologic brachytherapy demonstrate the ability to optimize tumor coverage and reduce radiation dose to normal tissues, resulting in improved outcomes for patients. In this article, we describe a methodology to aid applicator placement and treatment planning for 3 Tesla (3-T) MR-guided brachytherapy that was developed specifically for gynecologic cancers. This methodology has been used in 18 cases from September 2011 to May 2012 in the Advanced Multimodality Image Guided Operating (AMIGO) suite at Brigham and Women's Hospital. AMIGO comprises state-of-the-art tools for MR imaging, image analysis and treatment planning. An MR sequence using three-dimensional (3D)-balanced steady-state free precession in a 3-T MR scanner was identified as the best sequence for catheter identification with ballooning artifact at the tip. 3D treatment planning was performed using MR images. Items in development include software designed to support virtual needle trajectory planning that uses probabilistic bias correction, graph-based segmentation and image registration algorithms. The results demonstrate that 3-T MR image guidance has a role in gynecologic brachytherapy. These novel developments have the potential to improve targeted treatment to the tumor while sparing the normal tissues. PMID:22898699

  16. Malformations of cortical development: 3T magnetic resonance imaging features

    PubMed Central

    Battal, Bilal; Ince, Selami; Akgun, Veysel; Kocaoglu, Murat; Ozcan, Emrah; Tasar, Mustafa

    2015-01-01

    Malformation of cortical development (MCD) is a term representing an inhomogeneous group of central nervous system abnormalities, referring particularly to embriyological aspect as a consequence of any of the three developmental stages, i.e., cell proliferation, cell migration and cortical organization. These include cotical dysgenesis, microcephaly, polymicrogyria, schizencephaly, lissencephaly, hemimegalencephaly, heterotopia and focal cortical dysplasia. Since magnetic resonance imaging is the modality of choice that best identifies the structural anomalies of the brain cortex, we aimed to provide a mini review of MCD by using 3T magnetic resonance scanner images. PMID:26516429

  17. Travelling wave magnetic resonance imaging at 3 T

    NASA Astrophysics Data System (ADS)

    Vazquez, F.; Martin, R.; Marrufo, O.; Rodriguez, A. O.

    2013-08-01

    Waveguides have been successfully used to generate magnetic resonance images at 7 T with whole-body systems. The bore diameter limits the magnetic resonance signal transmitted because its specific cut-off frequency is greater than the majority of resonant frequencies in magnetic resonance imaging and spectroscopy. This restriction can be overcome by using a parallel-plate waveguide whose cut-off frequency is zero for the transverse electromagnetic modes and it can propagate any frequency. To study the potential benefits of travelling-wave excitation for whole-body imaging at 3 T, we compare numerical simulations of the principal mode propagation for a parallel-plate waveguide filled with a cylindrical phantom and two surface coils for all simulations at 1.5 T, 3 T, 4.7, 7 T, and 9.4 T. The principal mode shows very little variation of the field magnitude along the propagation direction at 3 T when compared to other higher resonant frequencies. Unlike the standard method for travelling-wave magnetic resonance imaging, a parallel-plate waveguide prototype was built and used together with a whole-body birdcage coil for signal transmission and a pair of circular coils for reception. Experimental B1 mapping was computed to investigate the feasibility of this approach and, the point spread function method was used to measure the imager performance. Human leg images were acquired to experimentally validate this approach. The numerical magnetic field and specific absorption rate of a simulated leg were computed and results are within the safety limits. The B1 mapping and point spread function results showed that it is possible to conduct travelling-wave imaging experiments with good imager performance. Human leg images were also obtained with the whole-body birdcage coil for comparison purposes. The simulated and in vivo travelling-wave results of the human leg correspond very well for the signal received. A similar image signal-to-noise ratio was observed for the

  18. Exome sequencing of case-unaffected-parents trios reveals recessive and de novo genetic variants in sporadic ALS

    PubMed Central

    Steinberg, Karyn Meltz; Yu, Bing; Koboldt, Daniel C.; Mardis, Elaine R.; Pamphlett, Roger

    2015-01-01

    The contribution of genetic variants to sporadic amyotrophic lateral sclerosis (ALS) remains largely unknown. Either recessive or de novo variants could result in an apparently sporadic occurrence of ALS. In an attempt to find such variants we sequenced the exomes of 44 ALS-unaffected-parents trios. Rare and potentially damaging compound heterozygous variants were found in 27% of ALS patients, homozygous recessive variants in 14% and coding de novo variants in 27%. In 20% of patients more than one of the above variants was present. Genes with recessive variants were enriched in nucleotide binding capacity, ATPase activity, and the dynein heavy chain. Genes with de novo variants were enriched in transcription regulation and cell cycle processes. This trio study indicates that rare private recessive variants could be a mechanism underlying some case of sporadic ALS, and that de novo mutations are also likely to play a part in the disease. PMID:25773295

  19. Disparate access to treatment regimens in chronic hepatitis C patients: data from the TRIO network.

    PubMed

    Younossi, Z M; Bacon, B R; Dieterich, D T; Flamm, S L; Kowdley, K; Milligan, S; Tsai, N; Nezam, A

    2016-06-01

    Despite the clinical success in the real-world of all oral hepatitis C virus (HCV) therapy with response rates approaching that seen in the clinical trials, access has been limited by many payers with discussion of prioritization of treatment based upon AASLD guidelines. We evaluated patients in the TRIO network who were prescribed sofosbuvir (SOF)-based regimens to determine reasons for not starting treatment. Trio Health is a disease management company that works in partnership with academic medical centres, community physicians and specialty pharmacies in the United States to optimize care for HCV. Data for 3841 patients prescribed a sofosbuvir-containing regimen between December 2013 and September 2014 were obtained through this programme. Of the entire group, 315 (8%) patients did not start the prescribed sofosbuvir-containing therapy. A total of 141 (45%) of the nonstart patients had a commercial plan as their primary insurance, 137 (44%) were primarily covered by Medicaid, 17 (5%) were primarily covered by Medicare, and 20 (6%) were either without coverage or coverage was not specified. Reasons for nonstarts were varied and overlapping. Only 15 patients (5% of nonstarts) did not start because they were unreachable or failed to complete required testing. Another 39 patients who did not start (12%) were following their physicians' direction to either wait for new treatment options or to hold treatment for an unspecified reason. Insurance-related processes and financial reasons accounted for 254 (81%) of the 315 nonstarts. The remaining 7 (2%) patients did not have a specified reason for not starting treatment. Nonstart rates were highest in the Medicaid-covered population at 35%. Medicare and Commercial nonstart rates were 2% and 6%, respectively. In a matched comparison, patients with commercial coverage were 6.5 times as likely to start SOF-based therapy compared to patients with Medicaid. Despite high SVR rates of SOF-based regimens in clinical practice

  20. Disease and Polygenic Architecture: Avoid Trio Design and Appropriately Account for Unscreened Control Subjects for Common Disease

    PubMed Central

    Peyrot, Wouter J.; Boomsma, Dorret I.; Penninx, Brenda W.J.H.; Wray, Naomi R.

    2016-01-01

    Genome-wide association studies (GWASs) are an optimal design for discovery of disease risk loci for diseases whose underlying genetic architecture includes many common causal loci of small effect (a polygenic architecture). We consider two designs that deserve careful consideration if the true underlying genetic architecture of the trait is polygenic: parent-offspring trios and unscreened control subjects. We assess these designs in terms of quantification of the total contribution of genome-wide genetic markers to disease risk (SNP heritability) and power to detect an associated risk allele. First, we show that trio designs should be avoided when: (1) the disease has a lifetime risk > 1%; (2) trio probands are ascertained from families with more than one affected sibling under which scenario the SNP heritability can drop by more than 50% and power can drop as much as from 0.9 to 0.15 for a sample of 20,000 subjects; or (3) assortative mating occurs (spouse correlation of the underlying liability to the disorder), which decreases the SNP heritability but not the power to detect a single locus in the trio design. Some studies use unscreened rather than screened control subjects because these can be easier to collect; we show that the estimated SNP heritability should then be scaled by dividing by (1 − K × u)2 for disorders with population prevalence K and proportion of unscreened control subjects u. When omitting to scale appropriately, the SNP heritability of, for example, major depressive disorder (K = 0.15) would be underestimated by 28% when none of the control subjects are screened. PMID:26849113

  1. 3 T magnetic resonance imaging of the musculoskeletal system.

    PubMed

    Guglielmi, G; Biccari, N; Mangano, F; Toffanin, R

    2010-06-01

    The increasing distribution of high-field (3 T) magnetic resonance (MR) systems for clinical use has been accompanied by the need to fully understand the advantages and disadvantages that the increase in signal quality confers. Continuous development of the coils is required to fully express the potential of these systems, especially given the synergy between parallel imaging and the recent multichannel phased-array coils, which are able to improve image quality, spatial resolution and diagnostic accuracy in musculoskeletal imaging. The increase in signal offered by the high field makes possible improved visualisation of bone, cartilage, tendons and ligaments. This advantage, together with increased spatial resolution, is particularly useful when studying joints or some of their components, the evaluation of which has produced suboptimal results in non arthrographic examinations such as the glenoid labrum of the shoulder and the articular cartilage of the knee. Thanks to the greater signal-to-noise ratio and improved spatial resolution, MR imaging at 3 T is able to notably increase diagnostic performance in the musculoskeletal setting, with a consequent improvement in patient treatment and management. PMID:20177987

  2. Cannabidiol promotes browning in 3T3-L1 adipocytes.

    PubMed

    Parray, Hilal Ahmad; Yun, Jong Won

    2016-05-01

    Recruitment of the brown-like phenotype in white adipocytes (browning) and activation of existing brown adipocytes are currently being investigated as a means to combat obesity. Thus, a wide variety of dietary agents that contribute to browning of white adipocytes have been identified. The present study was designed to investigate the effects of cannabidiol (CBD), a major nonpsychotropic phytocannabinoid of Cannabis sativa, on induction of browning in 3T3-L1 adipocytes. CBD enhanced expression of a core set of brown fat-specific marker genes (Ucp1, Cited1, Tmem26, Prdm16, Cidea, Tbx1, Fgf21, and Pgc-1α) and proteins (UCP1, PRDM16, and PGC-1α). Increased expression of UCP1 and other brown fat-specific markers contributed to the browning of 3T3-L1 adipocytes possibly via activation of PPARγ and PI3K. In addition, CBD increased protein expression levels of CPT1, ACSL, SIRT1, and PLIN while down-regulating JNK2, SREBP1, and LPL. These data suggest possible roles for CBD in browning of white adipocytes, augmentation of lipolysis, thermogenesis, and reduction of lipogenesis. In conclusion, the current data suggest that CBD plays dual modulatory roles in the form of inducing the brown-like phenotype as well as promoting lipid metabolism. Thus, CBD may be explored as a potentially promising therapeutic agent for the prevention of obesity. PMID:27067870

  3. A 3T Sodium and Proton Composite Array Breast Coil

    PubMed Central

    Kaggie, Joshua D.; Hadley, J. Rock; Badal, James; Campbell, John R.; Park, Daniel J.; Parker, Dennis L.; Morrell, Glen; Newbould, Rexford D.; Wood, Ali F.; Bangerter, Neal K.

    2013-01-01

    Purpose The objective of this study was to determine whether a sodium phased array would improve sodium breast MRI at 3T. The secondary objective was to create acceptable proton images with the sodium phased array in place. Methods A novel composite array for combined proton/sodium 3T breast MRI is compared to a coil with a single proton and sodium channel. The composite array consists of a 7-channel sodium receive array, a larger sodium transmit coil, and a 4-channel proton transceive array. The new composite array design utilizes smaller sodium receive loops than typically used in sodium imaging, uses novel decoupling methods between the receive loops and transmit loops, and uses a novel multi-channel proton transceive coil. The proton transceive coil reduces coupling between proton and sodium elements by intersecting the constituent loops to reduce their mutual inductance. The coil used for comparison consists of a concentric sodium and proton loop with passive decoupling traps. Results The composite array coil demonstrates a 2–5x improvement in SNR for sodium imaging and similar SNR for proton imaging when compared to a simple single-loop dual resonant design. Conclusion The improved SNR of the composite array gives breast sodium images of unprecedented quality in reasonable scan times. PMID:24105740

  4. Characterization of hyaluronate binding proteins isolated from 3T3 and murine sarcoma virus transformed 3T3 cells

    SciTech Connect

    Turley, E.A.; Moore, D.; Hayden, L.J.

    1987-06-02

    A hyaluronic acid binding fraction was purified from the supernatant media of both 3T3 and murine sarcoma virus (MSV) transformed 3T3 cultures by hyaluronate and immunoaffinity chromatography. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis resolved the hyaluronate affinity-purified fraction into three major protein bands of estimated molecular weight (M/sub r,e/) 70K, 66K, and 56K which contained hyaluronate binding activity and which were termed hyaluronate binding proteins (HABP). Hyaluronate affinity chromatography combined with immunoaffinity chromatography, using antibody directed against the larger HABP, allowed a 20-fold purification of HABP. Fractions isolated from 3T3 supernatant medium also contained additional binding molecules in the molecular weight range of 20K. This material was present in vanishingly small amounts and was not detected with a silver stain or with (/sup 35/S)methionine label. The three protein species isolated by hyaluronate affinity chromatography (M/sub r,e/ 70K, 66K, and 56K) were related to one another since they shared antigenic determinants and exhibited similar pI values. In isocratic conditions, HABP occurred as aggregates of up to 580 kilodaltons. Their glycoprotein nature was indicated by their incorporation of /sup 3/H-sugars. Enzyme-linked immunoadsorbent assay showed they were antigenically distinct from other hyaluronate binding proteins such as fibronectin, cartilage link protein, and the hyaluronate binding region of chondroitin sulfate proteoglycan. The results are discussed with regard both to the functional significance of hyaluronate-cell surface interactions in transformed as well as normal cells and to the relationship of HABP to other reported hyaluronate binding proteins.

  5. Performance evaluation of fluorescence tomography in a Siemens Inveon multimodality scanner

    NASA Astrophysics Data System (ADS)

    Lu, Yujie; Darne, Chinmay; Tan, I.-Chih; Zhu, Banghe; Rasmussen, John; Sevick-Muraca, Eva M.

    2014-05-01

    A tri-modal (PET/CT/Optical) small animal tomographic imaging system was developed by integrating our advanced non-contact intensified CCD (ICCD) frequency-domain fluorescence imaging components into a Siemens Inveon scanner. We performed a performance evaluation of the developed imaging system by using the developed regularization-free high-order radiative-transfer-based reconstruction algorithm and custom solid phantoms. Our results show that frequency-domain photon migration (FDPM) fluorescence tomography can achieve better tomographic images with less artifacts and more precise fluorescent source localization compared to the continuous-wave counterpart. The developed multimodal tomographic imaging system provides a powerful tool for translational biomedical research.

  6. The series of siemens SST-200 to SST-900 steam turbines in Russia

    NASA Astrophysics Data System (ADS)

    Kasilov, V. F.

    2015-04-01

    The chronology of cooperation between the transnational German concern Siemens and Russia is given. The designs of the SST-200-SST-900 series of industrial steam turbines for electrical capacities ranging from 10 to 180 MW that were installed at thermal power plants and industrial facilities of Russia in the period 2004-2014 are reviewed. The design features of the SST-600 steam turbine used as part of the PGU-200 combined-cycle plant installed at the Southwest cogeneration station are considered.

  7. Whole-exome sequencing in undiagnosed genetic diseases: interpreting 119 trios

    PubMed Central

    Zhu, Xiaolin; Petrovski, Slavé; Xie, Pingxing; Ruzzo, Elizabeth K.; Lu, Yi-Fan; McSweeney, K. Melodi; Ben-Zeev, Bruria; Nissenkorn, Andreea; Anikster, Yair; Oz-Levi, Danit; Dhindsa, Ryan S.; Hitomi, Yuki; Schoch, Kelly; Spillmann, Rebecca C.; Heimer, Gali; Marek-Yagel, Dina; Tzadok, Michal; Han, Yujun; Worley, Gordon; Goldstein, Jennifer; Jiang, Yong-Hui; Lancet, Doron; Pras, Elon; Shashi, Vandana; McHale, Duncan; Need, Anna C.; Goldstein, David B.

    2015-01-01

    Purpose: Despite the recognized clinical value of exome-based diagnostics, methods for comprehensive genomic interpretation remain immature. Diagnoses are based on known or presumed pathogenic variants in genes already associated with a similar phenotype. Here, we extend this paradigm by evaluating novel bioinformatics approaches to aid identification of new gene–disease associations. Genet Med 17 10, 774–781. Methods: We analyzed 119 trios to identify both diagnostic genotypes in known genes and candidate genotypes in novel genes. We considered qualifying genotypes based on their population frequency and in silico predicted effects we also characterized the patterns of genotypes enriched among this collection of patients. Genet Med 17 10, 774–781. Results: We obtained a genetic diagnosis for 29 (24%) of our patients. We showed that patients carried an excess of damaging de novo mutations in intolerant genes, particularly those shown to be essential in mice (P = 3.4 × 10−8). This enrichment is only partially explained by mutations found in known disease-causing genes. Genet Med 17 10, 774–781. Conclusion: This work indicates that the application of appropriate bioinformatics analyses to clinical sequence data can also help implicate novel disease genes and suggest expanded phenotypes for known disease genes. These analyses further suggest that some cases resolved by whole-exome sequencing will have direct therapeutic implications. Genet Med 17 10, 774–781. PMID:25590979

  8. Challenging the spin accumulation interpretation of local ``3T'' measurements

    NASA Astrophysics Data System (ADS)

    Tinkey, Holly; Appelbaum, Ian; Li, Pengke

    2015-03-01

    The recent observation of magnetoresistance in local ``three-terminal'' (3T) measurements on ferromagnet/ insulator/ semiconductor junctions have spawned many claims of direct bulk spin injection or ``accumulation''. We present a self-consistent model to rigorously calculate expected voltage changes due to electrochemical potential splitting from spin accumulation driven by pure elastic tunnel injection in such junctions, and find that the experimentally observed magnetoresistance vastly exceeds theoretical predictions in all doping, temperature, and bias voltage regimes. Our own experimental measurements using inelastic electron tunneling spectroscopy reveal that extrinsic impurities and defects within the junctions are responsible for the observed magnetoresistance signals, which cannot possibly be attributed to spin dephasing of polarized bulk electrons from elastic injection as claimed by proponents of the method. Supported by the Maryland Nanocenter, the Office of Naval Research Contract N000141110637, the National Science Foundation contracts ECCS-0901941 and ECCS-1231855 and Graduate Fellowship DGE13221, and the Defense Threat Reduction Agency contract HDTRA1.

  9. Uniform DT 3T burn: computations and sensitivities

    SciTech Connect

    Vold, Erik; Hryniw, Natalia; Hansen, Jon A; Kesler, Leigh A; Li, Frank

    2011-01-27

    A numerical model was developed in C to integrate the nonlinear deutrium-tritium (DT) burn equations in a three temperature (3T) approximation for spatially uniform test problems relevant to Inertial Confinement Fusion (ICF). Base model results are in excellent agreement with standard 3T results. Data from NDI, SESAME, and TOPS databases is extracted to create fits for the reaction rate parameter, the Planck opacity, and the coupling frequencies of the plasma temperatures. The impact of different fits (e.g., TOPS versus SESAME opacity data, higher order polynomial fits ofNDI data for the reaction rate parameter) were explored, and sensitivity to several model inputs are presented including: opacity data base, Coulomb logarithm, and Bremsstrahlung. Sensitivity to numerical integration time step size, and the relative insensitivity to the discretized numerics and numerical integration method was demonstrated. Variations in the IC for densities and temperatures were explored, showing similar DT burn profiles in most cases once ignition occurs. A coefficient multiplying the Compton coupling term (default, A = 1) can be adjusted to approximate results from more sophisticated models. The coefficient was reset (A = 0.4) to match the maximum temperatures resulting from standard multi-group simulations of the base case test problem. Setting the coefficient to a larger value, (A = 0.6) matches maximum ion temperatures in a kinetic simulation of a high density ICF-like regime. Matching peak temperatures does not match entire temperature-time profiles, indicating the Compton coefficient is density and time dependent as the photon distribution evolves. In the early time burn during the ignition of the DT, the present model with modified Compton coupling provides a very simple method to obtain a much improved match to the more accurate solution from the multi-group radiation model for these DT burn regimes.

  10. The 3T3-L1 adipocyte glycogen proteome

    PubMed Central

    2013-01-01

    Background Glycogen is a branched polysaccharide of glucose residues, consisting of α-1-4 glycosidic linkages with α-1-6 branches that together form multi-layered particles ranging in size from 30 nm to 300 nm. Glycogen spatial conformation and intracellular organization are highly regulated processes. Glycogen particles interact with their metabolizing enzymes and are associated with a variety of proteins that intervene in its biology, controlling its structure, particle size and sub-cellular distribution. The function of glycogen in adipose tissue is not well understood but appears to have a pivotal role as a regulatory mechanism informing the cells on substrate availability for triacylglycerol synthesis. To provide new molecular insights into the role of adipocyte glycogen we analyzed the glycogen-associated proteome from differentiated 3T3-L1-adipocytes. Results Glycogen particles from 3T3-L1-adipocytes were purified using a series of centrifugation steps followed by specific elution of glycogen bound proteins using α-1,4 glucose oligosaccharides, or maltodextrins, and tandem mass spectrometry. We identified regulatory proteins, 14-3-3 proteins, RACK1 and protein phosphatase 1 glycogen targeting subunit 3D. Evidence was also obtained for a regulated subcellular distribution of the glycogen particle: metabolic and mitochondrial proteins were abundant. Unlike the recently analyzed hepatic glycogen proteome, no endoplasmic proteins were detected, along with the recently described starch-binding domain protein 1. Other regulatory proteins which have previously been described as glycogen-associated proteins were not detected, including laforin, the AMPK beta-subunit and protein targeting to glycogen (PTG). Conclusions These data provide new molecular insights into the regulation of glycogen-bound proteins that are associated with the maintenance, organization and localization of the adipocyte glycogen particle. PMID:23521774

  11. Effect of filters and reconstruction algorithms on I-124 PET in Siemens Inveon PET scanner

    NASA Astrophysics Data System (ADS)

    Ram Yu, A.; Kim, Jin Su

    2015-10-01

    Purpose: To assess the effects of filtering and reconstruction on Siemens I-124 PET data. Methods: A Siemens Inveon PET was used. Spatial resolution of I-124 was measured to a transverse offset of 50 mm from the center FBP, 2D ordered subset expectation maximization (OSEM2D), 3D re-projection algorithm (3DRP), and maximum a posteriori (MAP) methods were tested. Non-uniformity (NU), recovery coefficient (RC), and spillover ratio (SOR) parameterized image quality. Mini deluxe phantom data of I-124 was also assessed. Results: Volumetric resolution was 7.3 mm3 from the transverse FOV center when FBP reconstruction algorithms with ramp filter was used. MAP yielded minimal NU with β =1.5. OSEM2D yielded maximal RC. SOR was below 4% for FBP with ramp, Hamming, Hanning, or Shepp-Logan filters. Based on the mini deluxe phantom results, an FBP with Hanning or Parzen filters, or a 3DRP with Hanning filter yielded feasible I-124 PET data.Conclusions: Reconstruction algorithms and filters were compared. FBP with Hanning or Parzen filters, or 3DRP with Hanning filter yielded feasible data for quantifying I-124 PET.

  12. Combining case-control and case-trio data from the same population in genetic association analyses: overview of approaches and illustration with a candidate gene study.

    PubMed

    Infante-Rivard, Claire; Mirea, Lucia; Bull, Shelley B

    2009-09-01

    In genetic association studies, investigators compare allele or genotype frequencies in unrelated case and control subjects or examine preferential allele transmissions from parents to affected offspring. In many genetic case-control studies, the collection of DNA material extends to relatives such as parents of cases. Thus, case-control and case-parent trio association analyses are possible. Whereas the goal of collecting genetic information from family members in a study initially designed as a case-control study is to enrich the genetic analysis, increase power, or address concern about population structure bias, methods of combining genetic data from unrelated case and control subjects with genetic trio data from the same study population are not well known. A number of hybrid approaches have been developed that utilize such data together. In this paper, the authors describe key features of genetic case-control and case-parent trio studies and review commonly used methods of genetic analysis for case-parent trio designs. In addition, they provide a pragmatic review of statistical methods and available software for existing hybrid approaches that combine various components of case-control and genetic trio data. The application of all methods is illustrated using a candidate gene study of childhood leukemia that included case-control subjects and their parents. PMID:19635737

  13. Whole-Genome Sequencing of a Canine Family Trio Reveals a FAM83G Variant Associated with Hereditary Footpad Hyperkeratosis.

    PubMed

    Sayyab, Shumaila; Viluma, Agnese; Bergvall, Kerstin; Brunberg, Emma; Jagannathan, Vidhya; Leeb, Tosso; Andersson, Göran; Bergström, Tomas F

    2016-03-01

    Over 250 Mendelian traits and disorders, caused by rare alleles have been mapped in the canine genome. Although each disease is rare in the dog as a species, they are collectively common and have major impact on canine health. With SNP-based genotyping arrays, genome-wide association studies (GWAS) have proven to be a powerful method to map the genomic region of interest when 10-20 cases and 10-20 controls are available. However, to identify the genetic variant in associated regions, fine-mapping and targeted resequencing is required. Here we present a new approach using whole-genome sequencing (WGS) of a family trio without prior GWAS. As a proof-of-concept, we chose an autosomal recessive disease known as hereditary footpad hyperkeratosis (HFH) in Kromfohrländer dogs. To our knowledge, this is the first time this family trio WGS-approach has been used successfully to identify a genetic variant that perfectly segregates with a canine disorder. The sequencing of three Kromfohrländer dogs from a family trio (an affected offspring and both its healthy parents) resulted in an average genome coverage of 9.2X per individual. After applying stringent filtering criteria for candidate causative coding variants, 527 single nucleotide variants (SNVs) and 15 indels were found to be homozygous in the affected offspring and heterozygous in the parents. Using the computer software packages ANNOVAR and SIFT to functionally annotate coding sequence differences, and to predict their functional effect, resulted in seven candidate variants located in six different genes. Of these, only FAM83G:c155G > C (p.R52P) was found to be concordant in eight additional cases, and 16 healthy Kromfohrländer dogs. PMID:26747202

  14. Whole-Genome Sequencing of a Canine Family Trio Reveals a FAM83G Variant Associated with Hereditary Footpad Hyperkeratosis

    PubMed Central

    Sayyab, Shumaila; Viluma, Agnese; Bergvall, Kerstin; Brunberg, Emma; Jagannathan, Vidhya; Leeb, Tosso; Andersson, Göran; Bergström, Tomas F.

    2016-01-01

    Over 250 Mendelian traits and disorders, caused by rare alleles have been mapped in the canine genome. Although each disease is rare in the dog as a species, they are collectively common and have major impact on canine health. With SNP-based genotyping arrays, genome-wide association studies (GWAS) have proven to be a powerful method to map the genomic region of interest when 10–20 cases and 10–20 controls are available. However, to identify the genetic variant in associated regions, fine-mapping and targeted resequencing is required. Here we present a new approach using whole-genome sequencing (WGS) of a family trio without prior GWAS. As a proof-of-concept, we chose an autosomal recessive disease known as hereditary footpad hyperkeratosis (HFH) in Kromfohrländer dogs. To our knowledge, this is the first time this family trio WGS-approach has been used successfully to identify a genetic variant that perfectly segregates with a canine disorder. The sequencing of three Kromfohrländer dogs from a family trio (an affected offspring and both its healthy parents) resulted in an average genome coverage of 9.2X per individual. After applying stringent filtering criteria for candidate causative coding variants, 527 single nucleotide variants (SNVs) and 15 indels were found to be homozygous in the affected offspring and heterozygous in the parents. Using the computer software packages ANNOVAR and SIFT to functionally annotate coding sequence differences, and to predict their functional effect, resulted in seven candidate variants located in six different genes. Of these, only FAM83G:c155G > C (p.R52P) was found to be concordant in eight additional cases, and 16 healthy Kromfohrländer dogs. PMID:26747202

  15. Dosimetric Characteristics of 6 MV Modified Beams by Physical Wedges of a Siemens Linear Accelerator.

    PubMed

    Zabihzadeh, Mansour; Birgani, Mohammad Javad Tahmasebi; Hoseini-Ghahfarokhi, Mojtaba; Arvandi, Sholeh; Hoseini, Seyed Mohammad; Fadaei, Mahbube

    2016-01-01

    Physical wedges still can be used as missing tissue compensators or filters to alter the shape of isodose curves in a target volume to reach an optimal radiotherapy plan without creating a hotspot. The aim of this study was to investigate the dosimetric properties of physical wedges filters such as off-axis photon fluence, photon spectrum, output factor and half value layer. The photon beam quality of a 6 MV Primus Siemens modified by 150 and 450 physical wedges was studied with BEAMnrc Monte Carlo (MC) code. The calculated present depth dose and dose profile curves for open and wedged photon beam were in good agreement with the measurements. Increase of wedge angle increased the beam hardening and this effect was more pronounced at the heal region. Using such an accurate MC model to determine of wedge factors and implementation of it as a calculation algorithm in the future treatment planning systems is recommended. PMID:27221838

  16. GEANT4 Application for the Simulation of the Head of a Siemens Primus Linac

    SciTech Connect

    Cortes-Giraldo, M. A.; Quesada, J. M.; Gallardo, M. I.

    2010-04-26

    The Monte Carlo simulation of the head of a Siemens Primus Linac used at Virgen Macarena Hospital (Sevilla, Spain) has been performed using the code GEANT4, version 9.2. In this work, the main features of the application built by our group are presented. They are mainly focused in the optimization of the performance of the simulation. The geometry, including the water phantom, has been entirely wrapped by a shielding volume which discards all the particles escaping far away through its walls. With this, a factor of four in the time spent by the simulation can be saved. An interface to read and write phase-space files in IAEA format has been also developed to save CPU time in our simulations. Finally, some calculations of the dose absorption in the water phantom have been done and compared with the results given by EGSnrc and with experimental data obtained for the calibration of the machine.

  17. Siemens solar CIS photovoltaic module and system performance at the National Renewable Energy Laboratory

    SciTech Connect

    Strand, T.; Kroposki, B.; Hansen, R.; Willett, D.

    1996-05-01

    This paper evaluates the individual module and array performance of Siemens Solar Industries copper indium diselenide (CIS) polycrystalline thin-film technology. This is accomplished by studying module and array performance over time. Preliminary temperature coefficients for maximum power, maximum-power voltage, maximum-power current, open-circuit voltage, short-circuit current, and fill factor are determined at both the module and array level. These coefficients are used to correct module/array performance to 25{degrees}C to evaluate stability. The authors show that CIS exhibits a strong inverse correlation between array power and back-of-module temperature. This is due mainly to the narrow bandgap of the CIS material, which results in a strong inverse correlation between voltage and temperature. They also show that the temperature-corrected module and array performance has been relatively stable over the evaluation interval ({approx}2 years).

  18. Meta-analysis of 2,104 trios provides support for 10 new genes for intellectual disability.

    PubMed

    Lelieveld, Stefan H; Reijnders, Margot R F; Pfundt, Rolph; Yntema, Helger G; Kamsteeg, Erik-Jan; de Vries, Petra; de Vries, Bert B A; Willemsen, Marjolein H; Kleefstra, Tjitske; Löhner, Katharina; Vreeburg, Maaike; Stevens, Servi J C; van der Burgt, Ineke; Bongers, Ernie M H F; Stegmann, Alexander P A; Rump, Patrick; Rinne, Tuula; Nelen, Marcel R; Veltman, Joris A; Vissers, Lisenka E L M; Brunner, Han G; Gilissen, Christian

    2016-09-01

    To identify candidate genes for intellectual disability, we performed a meta-analysis on 2,637 de novo mutations, identified from the exomes of 2,104 patient-parent trios. Statistical analyses identified 10 new candidate ID genes: DLG4, PPM1D, RAC1, SMAD6, SON, SOX5, SYNCRIP, TCF20, TLK2 and TRIP12. In addition, we show that these genes are intolerant to nonsynonymous variation and that mutations in these genes are associated with specific clinical ID phenotypes. PMID:27479843

  19. Clinical experience with shock-wave lithotripsy using the Siemens Modularis Vario lithotripter

    PubMed Central

    Hassouna, Mohamed E.; Oraby, Samir; Sameh, Wael; El-Abbady, Ahmed

    2011-01-01

    Purpose To assess the effectiveness of a lithotripter (Modularis Vario; Siemens, AG Healthcare, Munich, Germany) in the management of renal and ureteric stones. Patients and methods In all, 1146 adult patients with renal or ureteric stones were treated at one urological centre using the latest model of the Modularis Vario lithotripter. The effectiveness of lithotripsy and re-treatment rate were assessed. Data were obtained on stone location, stone size, shock wave usage, success rate, and complications. Results Between May 2007 and November 2009, 698 patients with renal stones and 448 with ureteric stones underwent extracorporeal shock-wave lithotripsy (ESWL). The mean (SD) renal stone size was 12.8 (3.8) mm; a mean of 1.36 sessions was required, with a mean (SD) number of 3744 (1961) shocks delivered per renal stone. After 3 months, the success rate defined as the patient being stone-free or with residual fragments of <4 mm; for renal stones the rate was 91.1%, with a 6.9% complication rate in the form of steinstrasse and severe renal colic. The mean (SD) ureteric stone size was 10.4 (2.7) mm. A mean of 1.37 sessions was required, with a mean (SD) of 4551 (2467) shocks delivered for each ureteric stone. The success rate for ureteric stones was 89.5%, with a 5.6% complication rate. The overall efficiency quotient was 0.66. Conclusion The Siemens Modularis Vario lithotripter is a safe and effective machine for treating renal and ureteric stones. PMID:26579276

  20. Modeling and simulation of silicon epitaxial growth in Siemens CVD reactor

    NASA Astrophysics Data System (ADS)

    Ni, Haoyin; Lu, Shijie; Chen, Caixia

    2014-10-01

    Siemens CVD reactor is an important chemical device for the production of polysilicon. The chemical and physical phenomenon involved in the reactor is very complex. Understanding the multispecies thermal fluid transport and its interaction with the gas/surface reactions is crucial for an optimal design and operation of the reactor. In the present paper, a mathematical model was constructed to describe the fluid dynamics, the heat and mass transfer and the reaction kinetics of the epitaxial growth process in industrial CVD reactors. A modified reaction kinetics model was used to represent the gas phase and surface reactions. The kinetics model was validated using the published experimental data obtained in a temperature range similar to the industrial CVD processes of silicon productions. The epitaxial growth of silicon in a Siemens reactor was simulated using commercial Computational Fluid Dynamics (CFD) software ANSYS FLUENT. The distributions of gas velocity, temperature and species concentrations in the reactor were predicted numerically. Based on the numerical simulation results, a sensitivity analysis was carried out to determine the key factors influencing the growth rate in industrial CVD reactors. Under the conditions of fixed heating power applied to three different rod diameters of 50 mm, 80 mm and 100 mm, the simulated results show, when the rods' diameter is 50 mm, the surface temperature is high and the gas temperature is low, the growth rate of silicon is determined by the transport of gas species. When the rods' diameter increases to 80 mm, the averaged surface temperature decreases to 1361 K, the surface reaction rate and transport of gas species control the growth rate of Si together. When the rods' diameter is 100 mm, the surface temperature decreases further, the rates of surface reactions become the control factor of deposition rate of Si.

  1. 26 CFR 54.9802-3T - Additional requirements prohibiting discrimination based on genetic information (temporary).

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 26 Internal Revenue 17 2014-04-01 2014-04-01 false Additional requirements prohibiting discrimination based on genetic information (temporary). 54.9802-3T Section 54.9802-3T Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY (CONTINUED) MISCELLANEOUS EXCISE TAXES (CONTINUED) PENSION EXCISE TAXES § 54.9802-3T...

  2. 26 CFR 54.9802-3T - Additional requirements prohibiting discrimination based on genetic information (temporary).

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 26 Internal Revenue 17 2012-04-01 2012-04-01 false Additional requirements prohibiting discrimination based on genetic information (temporary). 54.9802-3T Section 54.9802-3T Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY (CONTINUED) MISCELLANEOUS EXCISE TAXES (CONTINUED) PENSION EXCISE TAXES § 54.9802-3T...

  3. 26 CFR 54.9802-3T - Additional requirements prohibiting discrimination based on genetic information (temporary).

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 26 Internal Revenue 17 2013-04-01 2013-04-01 false Additional requirements prohibiting discrimination based on genetic information (temporary). 54.9802-3T Section 54.9802-3T Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY (CONTINUED) MISCELLANEOUS EXCISE TAXES (CONTINUED) PENSION EXCISE TAXES § 54.9802-3T...

  4. 26 CFR 1.1471-3T - Identification of payee (temporary).

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 26 Internal Revenue 12 2014-04-01 2014-04-01 false Identification of payee (temporary). 1.1471-3T Section 1.1471-3T Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY (CONTINUED) INCOME TAX (CONTINUED) INCOME TAXES (CONTINUED) Information Reporting by Foreign Financial Institutions § 1.1471-3T Identification of payee...

  5. 49 CFR 178.45 - Specification 3T seamless steel cylinder.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 49 Transportation 3 2011-10-01 2011-10-01 false Specification 3T seamless steel cylinder. 178.45... PACKAGINGS Specifications for Cylinders § 178.45 Specification 3T seamless steel cylinder. (a) Type, size, and service pressure. A DOT 3T cylinder is a seamless steel cylinder with a minimum water capacity...

  6. 49 CFR 178.45 - Specification 3T seamless steel cylinder.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 49 Transportation 3 2013-10-01 2013-10-01 false Specification 3T seamless steel cylinder. 178.45... PACKAGINGS Specifications for Cylinders § 178.45 Specification 3T seamless steel cylinder. (a) Type, size, and service pressure. A DOT 3T cylinder is a seamless steel cylinder with a minimum water capacity...

  7. 49 CFR 178.45 - Specification 3T seamless steel cylinder.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 49 Transportation 3 2014-10-01 2014-10-01 false Specification 3T seamless steel cylinder. 178.45... PACKAGINGS Specifications for Cylinders § 178.45 Specification 3T seamless steel cylinder. (a) Type, size, and service pressure. A DOT 3T cylinder is a seamless steel cylinder with a minimum water capacity...

  8. 49 CFR 178.45 - Specification 3T seamless steel cylinder.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 49 Transportation 3 2012-10-01 2012-10-01 false Specification 3T seamless steel cylinder. 178.45... PACKAGINGS Specifications for Cylinders § 178.45 Specification 3T seamless steel cylinder. (a) Type, size, and service pressure. A DOT 3T cylinder is a seamless steel cylinder with a minimum water capacity...

  9. 26 CFR 1.892-3T - Income of foreign governments (temporary regulations).

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 26 Internal Revenue 9 2011-04-01 2011-04-01 false Income of foreign governments (temporary regulations). 1.892-3T Section 1.892-3T Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY (CONTINUED) INCOME TAX (CONTINUED) INCOME TAXES (CONTINUED) Miscellaneous Provisions § 1.892-3T Income of foreign governments...

  10. 26 CFR 1.702-3T - 4-Year spread (temporary).

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 26 Internal Revenue 8 2011-04-01 2011-04-01 false 4-Year spread (temporary). 1.702-3T Section 1.702-3T Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY (CONTINUED) INCOME TAX (CONTINUED) INCOME TAXES (CONTINUED) Partners and Partnerships § 1.702-3T 4-Year spread (temporary). (a) Applicability. This section applies to...

  11. 26 CFR 1.25-3T - Qualified mortgage credit certificate (Temporary).

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 26 Internal Revenue 1 2012-04-01 2012-04-01 false Qualified mortgage credit certificate (Temporary). 1.25-3T Section 1.25-3T Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY INCOME TAX INCOME TAXES Changes in Rates During A Taxable Year § 1.25-3T Qualified mortgage credit certificate (Temporary). (a) Definition of...

  12. 26 CFR 1.108(i)-3T - Rules for the deduction of OID (temporary).

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ....108(i)-3T Section 1.108(i)-3T Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY....108(i)-3T Rules for the deduction of OID (temporary). (a) Deemed debt-for-debt exchanges—(1) In general. For purposes of section 108(i)(2) (relating to deferred OID deductions that arise in certain...

  13. 26 CFR 1.103(n)-3T - Private activity bond limit (temporary).

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 26 Internal Revenue 2 2010-04-01 2010-04-01 false Private activity bond limit (temporary). 1.103(n)-3T Section 1.103(n)-3T Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY....103(n)-3T Private activity bond limit (temporary). Q-1: What is the “State ceiling”? A-1: In...

  14. 26 CFR 1.103(n)-3T - Private activity bond limit (temporary).

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 26 Internal Revenue 2 2011-04-01 2011-04-01 false Private activity bond limit (temporary). 1.103(n)-3T Section 1.103(n)-3T Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY....103(n)-3T Private activity bond limit (temporary). Q-1: What is the “State ceiling”? A-1: In...

  15. 26 CFR 1.103(n)-3T - Private activity bond limit (temporary).

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 26 Internal Revenue 2 2013-04-01 2013-04-01 false Private activity bond limit (temporary). 1.103(n)-3T Section 1.103(n)-3T Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY....103(n)-3T Private activity bond limit (temporary). Q-1: What is the “State ceiling”? A-1: In...

  16. 26 CFR 1.103(n)-3T - Private activity bond limit (temporary).

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 26 Internal Revenue 2 2014-04-01 2014-04-01 false Private activity bond limit (temporary). 1.103(n)-3T Section 1.103(n)-3T Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY....103(n)-3T Private activity bond limit (temporary). Q-1: What is the “State ceiling”? A-1: In...

  17. 26 CFR 1.103(n)-3T - Private activity bond limit (temporary).

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 26 Internal Revenue 2 2012-04-01 2012-04-01 false Private activity bond limit (temporary). 1.103(n)-3T Section 1.103(n)-3T Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY....103(n)-3T Private activity bond limit (temporary). Q-1: What is the “State ceiling”? A-1: In...

  18. 26 CFR 1.921-3T - Temporary regulations; Foreign sales corporation general rules.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 26 Internal Revenue 10 2010-04-01 2010-04-01 false Temporary regulations; Foreign sales corporation general rules. 1.921-3T Section 1.921-3T Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY (CONTINUED) INCOME TAX (CONTINUED) INCOME TAXES Earned Income of Citizens of United States § 1.921-3T Temporary regulations;...

  19. 26 CFR 1.132-3T - Qualified employee discount-1985 through 1988 (temporary).

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... (temporary). 1.132-3T Section 1.132-3T Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY (CONTINUED) INCOME TAX (CONTINUED) INCOME TAXES (CONTINUED) Items Specifically Excluded from Gross Income § 1.132-3T Qualified employee discount—1985 through 1988 (temporary). (a) In general—(1) Definition....

  20. 26 CFR 1.132-3T - Qualified employee discount-1985 through 1988 (temporary).

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... (temporary). 1.132-3T Section 1.132-3T Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY (CONTINUED) INCOME TAX (CONTINUED) INCOME TAXES (CONTINUED) Items Specifically Excluded from Gross Income § 1.132-3T Qualified employee discount—1985 through 1988 (temporary). (a) In general—(1) Definition....

  1. Removal of Microbial Contaminants in Drinking Water: Siemens Corporation Memcor® L10V Ultrafiltration Module

    EPA Science Inventory

    The Siemens Memcor L10V UF module was tested for removal of Cryptosporidium parvum oocyste, endospores of the bacteria Bacillus atrophaeus, and the MS2 coliphage virus according to the requirements of the EPA Long-Term 2 Enchanced Surface Water Treatment Rule (LT2ESWTR). Five m...

  2. Removal of Microbial Contaminants in Drinking Water: Siemens Corporation Memcor® L20V Ultrafiltration Module

    EPA Science Inventory

    The Siemens Memcor L20V UF module was tested for removal of endospores of the bacteria Bacillus atrophaeus and the MS2 coliphage virus according to the requirements of the EPA Long-Term 2 Enchanced Surface Water Treatment Rule (LT2ESWTR). B. atrophaeus served as a surrogate for ...

  3. Removal of Microbial Contaminants in Drinking Water: Siemens Corporation Memcor® S10V Ultrafiltration Module

    EPA Science Inventory

    The Siemens Memcor S10V UF module was tested for removal of endospores of the bacteria Bacillus atrophaeus and the MS2 coliphage virus according to the requirements of the EPA Long-Term 2 Enchanced Surface Water Treatment Rule (LT2ESWTR). B. atrophaeus served as a surrogate for ...

  4. Trio of Stellar Occultations by Pluto One Year Prior to New Horizons' Arrival

    NASA Astrophysics Data System (ADS)

    Pasachoff, Jay M.; Person, Michael J.; Bosh, Amanda S.; Sickafoose, Amanda A.; Zuluaga, Carlos; Kosiarek, Molly R.; Levine, Stephen E.; Osip, David J.; Schiff, Avery; Seeger, Christina H.; Babcock, Bryce A.; Rojo, Patricio; Servajean, Elise

    2016-04-01

    We observed occultations by Pluto during a predicted series of events in 2014 July with the 1 m telescope of the Mt. John Observatory in New Zealand. The predictions were based on updated astrometry obtained in the previous months at the USNO, CTIO, and Lowell Observatories. We successfully detected occultations by Pluto of an R = 18 mag star on July 23 (14:23:32 ± 00:00:04 UTC to 14:25:30 ± 00:00:04 UTC), with a drop of 75% of the unocculted stellar signal, and of an R = 17 star on July 24 (11:41:30 ± 00:00:08 UTC to 11:43:28 ± 00:00:08 UTC), with a drop of 80% of the unocculted stellar signal, both with 20 s exposures with our frame-transfer Portable Occultation, Eclipse, and Transit System. Since Pluto had a geocentric velocity of 22.51 km s-1 on July 23 and 22.35 km s-1 on July 24, these intervals yield limits on the chord lengths (surface and lower atmosphere) of 2700 ± 130 km and 2640 ± 250 km, respectively, indicating that the events were near central, and therefore provide astrometric constraints on the prediction method. Our coordinated observations with the 4 m AAT in Australia on July 23 and the 6.5 m Magellan/Clay on Las Campanas, the 4.1 m Southern Astrophysical Research Telescope on Cerro Pachön, the 2.5 m DuPont on Las Campanas (LCO), the 0.6 m SARA-South on Cerro Tololo of the Southeastern Association for Research in Astronomy (SARA), the MPI/ESO 2.2 m on La Silla, and the 0.45 m Cerro Calán telescope and 0.36 telescope in Constitución in Chile on July 27 and 31, which would have provided higher-cadence observations for studies of Pluto’s atmosphere, were largely foiled by clouds, but led to detection with the LCO Magellan/Clay and DuPont Telescopes on July 31 of the grazing occultation of a previously unknown 15th-magnitude star, completing the trio of occultations successfully observed and reported in this paper.

  5. Posttraumatic Stress Disorder: Structural Characterization with 3-T MR Imaging.

    PubMed

    Li, Shiguang; Huang, Xiaoqi; Li, Lingjiang; Du, Fei; Li, Jing; Bi, Feng; Lui, Su; Turner, Jessica A; Sweeney, John A; Gong, Qiyong

    2016-08-01

    Purpose To explore cerebral alterations related to the emergence of posttraumatic stress disorder (PTSD) by using three-dimensional T1-weighted imaging and also to explore the relationship of gray and white matter abnormalities and the anatomic changes with clinical severity and duration of time since the trauma. Materials and Methods Informed consent was provided, and the prospective study was approved by the ethics committee of the West China Hospital. Recruited were 67 patients with PTSD and 78 adult survivors without PTSD 7-15 months after a devastating earthquake in western China. All participants underwent magnetic resonance (MR) imaging with a 3-T imager to obtain anatomic images. Cortical thickness and volumes of 14 subcortical gray matter structures and five subregions of the corpus callosum were analyzed with software. Statistical differences between patients with PTSD and healthy survivors were evaluated with a general linear model. Averaged data from the regions with volumetric or cortical thickness differences between groups were extracted in each individual to examine correlations between morphometric measures and clinical profiles. Results Patients with PTSD showed greater cortical thickness in the right superior temporal gyrus, inferior parietal lobule, and left precuneus (P < .05; Monte Carlo null-z simulation corrected) and showed reduced volume in the posterior portion of the corpus callosum (F = 6.167; P = .014) compared with healthy survivors of the earthquake. PTSD severity was positively correlated with cortical thickness in the left precuneus (r = 0.332; P = .008). The volumes of posterior corpus callosum were negatively correlated with PTSD ratings in all survivors (r = -0.210; P = .013) and with cortical thickness of the left precuneus in patients with PTSD (r = -0.302; P = .017). Conclusion Results indicate that patients with PTSD had alterations in both cerebral gray matter and white matter compared with individuals who experienced

  6. Multi-Parametric Neuroimaging Reproducibility: A 3T Resource Study

    PubMed Central

    Landman, Bennett A.; Huang, Alan J.; Gifford, Aliya; Vikram, Deepti S.; Lim, Issel Anne L.; Farrell, Jonathan A.D.; Bogovic, John A.; Hua, Jun; Chen, Min; Jarso, Samson; Smith, Seth A.; Joel, Suresh; Mori, Susumu; Pekar, James J.; Barker, Peter B.; Prince, Jerry L.; van Zijl, Peter C.M.

    2010-01-01

    Modern MRI image processing methods have yielded quantitative, morphometric, functional, and structural assessments of the human brain. These analyses typically exploit carefully optimized protocols for specific imaging targets. Algorithm investigators have several excellent public data resources to use to test, develop, and optimize their methods. Recently, there has been an increasing focus on combining MRI protocols in multi-parametric studies. Notably, these have included innovative approaches for fusing connectivity inferences with functional and/or anatomical characterizations. Yet, validation of the reproducibility of these interesting and novel methods has been severely hampered by the limited availability of appropriate multi-parametric data. We present an imaging protocol optimized to include state-of-the-art assessment of brain function, structure, micro-architecture, and quantitative parameters within a clinically feasible 60 minute protocol on a 3T MRI scanner. We present scan-rescan reproducibility of these imaging contrasts based on 21 healthy volunteers (11 M/10 F, 22–61 y/o). The cortical gray matter, cortical white matter, ventricular cerebrospinal fluid, thalamus, putamen, caudate, cerebellar gray matter, cerebellar white matter, and brainstem were identified with mean volume-wise reproducibility of 3.5%. We tabulate the mean intensity, variability and reproducibility of each contrast in a region of interest approach, which is essential for prospective study planning and retrospective power analysis considerations. Anatomy was highly consistent on structural acquisition (~1–5% variability), while variation on diffusion and several other quantitative scans was higher (~<10%). Some sequences are particularly variable in specific structures (ASL exhibited variation of 28% in the cerebral white matter) or in thin structures (quantitative T2 varied by up to 73% in the caudate) due, in large part, to variability in automated ROI placement. The

  7. Construction of a Thermal Vacuum Chamber for Environment Test of Triple CubeSat Mission TRIO-CINEMA

    NASA Astrophysics Data System (ADS)

    Jeon, Jeheon; Lee, Seongwhan; Yoon, Seyoung; Seon, Jongho; Jin, Ho; Lee, Donghun; Lin, Robert P.

    2013-12-01

    TRiplet Ionospheric Observatory-CubeSat for Ion, Neutron, Electron & MAgnetic fields (TRIO-CINEMA) is a CubeSat with 3.14 kg in weight and 3-U (10 × 10 × 30 cm) in size, jointly developed by Kyung Hee University and UC Berkeley to measure magnetic fields of near Earth space and detect plasma particles. When a satellite is launched into orbit, it encounters ultrahigh vacuum and extreme temperature. To verify the operation and survivability of the satellite in such an extreme space environment, experimental tests are conducted on the ground using thermal vacuum chamber. This paper describes the temperature control device and monitoring system suitable for CubeSat test environment using the thermal vacuum chamber of the School of Space Research, Kyung Hee University. To build the chamber, we use a general purpose thermal analysis program and NX 6.0 TMG program. We carry out thermal vacuum tests on the two flight models developed by Kyung Hee University based on the thermal model of the TRIO-CINEMA satellite. It is expected from this experiment that proper operation of the satellite in the space environment will be achieved.

  8. Lysophosphatidic acid receptor-5 negatively regulates cellular responses in mouse fibroblast 3T3 cells

    SciTech Connect

    Dong, Yan; Hirane, Miku; Araki, Mutsumi; Fukushima, Nobuyuki; Tsujiuchi, Toshifumi

    2014-04-04

    Highlights: • LPA{sub 5} inhibits the cell growth and motile activities of 3T3 cells. • LPA{sub 5} suppresses the cell motile activities stimulated by hydrogen peroxide in 3T3 cells. • Enhancement of LPA{sub 5} on the cell motile activities inhibited by LPA{sub 1} in 3T3 cells. • The expression and activation of Mmp-9 were inhibited by LPA{sub 5} in 3T3 cells. • LPA signaling via LPA{sub 5} acts as a negative regulator of cellular responses in 3T3 cells. - Abstract: Lysophosphatidic acid (LPA) signaling via G protein-coupled LPA receptors (LPA{sub 1}–LPA{sub 6}) mediates a variety of biological functions, including cell migration. Recently, we have reported that LPA{sub 1} inhibited the cell motile activities of mouse fibroblast 3T3 cells. In the present study, to evaluate a role of LPA{sub 5} in cellular responses, Lpar5 knockdown (3T3-L5) cells were generated from 3T3 cells. In cell proliferation assays, LPA markedly stimulated the cell proliferation activities of 3T3-L5 cells, compared with control cells. In cell motility assays with Cell Culture Inserts, the cell motile activities of 3T3-L5 cells were significantly higher than those of control cells. The activity levels of matrix metalloproteinases (MMPs) were measured by gelatin zymography. 3T3-L5 cells stimulated the activation of Mmp-2, correlating with the expression levels of Mmp-2 gene. Moreover, to assess the co-effects of LPA{sub 1} and LPA{sub 5} on cell motile activities, Lpar5 knockdown (3T3a1-L5) cells were also established from Lpar1 over-expressing (3T3a1) cells. 3T3a1-L5 cells increased the cell motile activities of 3T3a1 cells, while the cell motile activities of 3T3a1 cells were significantly lower than those of control cells. These results suggest that LPA{sub 5} may act as a negative regulator of cellular responses in mouse fibroblast 3T3 cells, similar to the case for LPA{sub 1}.

  9. Release and disposal of materials during decommissioning of Siemens MOX fuel fabrication plant at Hanau, Germany

    SciTech Connect

    Koenig, Werner; Baumann, Roland

    2007-07-01

    In September 2006, decommissioning and dismantling of the Siemens MOX Fuel Fabrication Plant in Hanau were completed. The process equipment and the fabrication buildings were completely decommissioned and dismantled. The other buildings were emptied in whole or in part, although they were not demolished. Overall, the decommissioning process produced approximately 8500 Mg of radioactive waste (including inactive matrix material); clearance measurements were also performed for approximately 5400 Mg of material covering a wide range of types. All the equipment in which nuclear fuels had been handled was disposed of as radioactive waste. The radioactive waste was conditioned on the basis of the requirements specified for the projected German final disposal site 'Schachtanlage Konrad'. During the pre-conditioning, familiar processes such as incineration, compacting and melting were used. It has been shown that on account of consistently applied activity containment (barrier concept) during operation and dismantling, there has been no significant unexpected contamination of the plant. Therefore almost all the materials that were not a priori destined for radioactive waste were released without restriction on the basis of the applicable legal regulations (chap. 29 of the Radiation Protection Ordinance), along with the buildings and the plant site. (authors)

  10. Sensitivity analysis of an asymmetric Monte Carlo beam model of a Siemens Primus accelerator.

    PubMed

    Schreiber, Eric C; Sawkey, Daren L; Faddegon, Bruce A

    2012-01-01

    The assumption of cylindrical symmetry in radiotherapy accelerator models can pose a challenge for precise Monte Carlo modeling. This assumption makes it difficult to account for measured asymmetries in clinical dose distributions. We have performed a sensitivity study examining the effect of varying symmetric and asymmetric beam and geometric parameters of a Monte Carlo model for a Siemens PRIMUS accelerator. The accelerator and dose output were simulated using modified versions of BEAMnrc and DOSXYZnrc that allow lateral offsets of accelerator components and lateral and angular offsets for the incident electron beam. Dose distributions were studied for 40 × 40 cm² fields. The resulting dose distributions were analyzed for changes in flatness, symmetry, and off-axis ratio (OAR). The electron beam parameters having the greatest effect on the resulting dose distributions were found to be electron energy and angle of incidence, as high as 5% for a 0.25° deflection. Electron spot size and lateral offset of the electron beam were found to have a smaller impact. Variations in photon target thickness were found to have a small effect. Small lateral offsets of the flattening filter caused significant variation to the OAR. In general, the greatest sensitivity to accelerator parameters could be observed for higher energies and off-axis ratios closer to the central axis. Lateral and angular offsets of beam and accelerator components have strong effects on dose distributions, and should be included in any high-accuracy beam model. PMID:22402376

  11. GEANT4 Application for the Simulation of the Head of a Siemens Primus Linac

    NASA Astrophysics Data System (ADS)

    Cortés-Giraldo, M. A.; Quesada, J. M.; Gallardo, M. I.

    2010-04-01

    The Monte Carlo simulation of the head of a Siemens Primus Linac used at Virgen Macarena Hospital (Sevilla, Spain) has been performed using the code GEANT4 [1-2], version 9.2. In this work, the main features of the application built by our group are presented. They are mainly focused in the optimization of the performance of the simulation. The geometry, including the water phantom, has been entirely wrapped by a shielding volume which discards all the particles escaping far away through its walls. With this, a factor of four in the time spent by the simulation can be saved. An interface to read and write phase-space files in IAEA format has been also developed to save CPU time in our simulations [3-4]. Finally, some calculations of the dose absorption in the water phantom have been done and compared with the results given by EGSnrc [5] and with experimental data obtained for the calibration of the machine.

  12. The measurement of photoneutrons in the vicinity of a Siemens Primus linear accelerator.

    PubMed

    Lin, J P; Chu, T C; Lin, S Y; Liu, M T

    2001-09-01

    This study involves the measurement of photoneutron contamination emitted from a Siemens Primus medical linear accelerator by using BD-PND bubble detectors. Various bubble detectors were arranged around the linac head with the interval of I m and at the same height as the isocenter to measure the dose equivalent distribution in the treatment room. The measurements were performed for 15 MV X-rays with 40 x 40cm2 and 0 x 0cm2 fields and for 15,18, and 21 MeV electrons with 25 x 25 cm2 electron cone. Neutron dose equivalent rate at the points of measurement in the treatment room decreased with increasing distance to the isocenter. The maximum neutron dose equivalents were at the isocenter, and the values for 15MV 40 x 40 and 0 x 0 cm2 were 1843+/-90 and 169.9+/-59.9 microSv per Gy X-ray, respectively. The values for 15, 18 and 21 MeV electrons with 25 x 25 cm2 cones were 100.0+/-20.4, 262.7+/-61.2 and 349.0+/-29.6 microSv per Gy electron, respectively. The neutron contamination of electrons less than 12 MeV was below the detection limit. PMID:11515653

  13. 26 CFR 1.444-3T - Manner and time of making section 444 election (temporary).

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 26 Internal Revenue 6 2012-04-01 2012-04-01 false Manner and time of making section 444 election (temporary). 1.444-3T Section 1.444-3T Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY (CONTINUED) INCOME TAX (CONTINUED) INCOME TAXES (CONTINUED) Accounting Periods § 1.444-3T Manner and time of making section 444 election...

  14. Investigation of the mechanical performance of Siemens linacs components during arc: gantry, MLC, and electronic portal imaging device

    PubMed Central

    Rowshanfarzad, Pejman; Häring, Peter; Riis, Hans L; Zimmermann, Sune J; Ebert, Martin A

    2015-01-01

    Background In radiotherapy treatments, it is crucial to monitor the performance of linac components including gantry, collimation system, and electronic portal imaging device (EPID) during arc deliveries. In this study, a simple EPID-based measurement method is suggested in conjunction with an algorithm to investigate the stability of these systems at various gantry angles with the aim of evaluating machine-related errors in treatments. Methods The EPID sag, gantry sag, changes in source-to-detector distance (SDD), EPID and collimator skewness, EPID tilt, and the sag in leaf bank assembly due to linac rotation were separately investigated by acquisition of 37 EPID images of a simple phantom with five ball bearings at various gantry angles. A fast and robust software package was developed for automated analysis of image data. Three Siemens linacs were investigated. Results The average EPID sag was within 1 mm for all tested linacs. Two machines showed >1 mm gantry sag. Changes in the SDD values were within 7.5 mm. EPID skewness and tilt values were <1° in all machines. The maximum sag in leaf bank assembly was <1 mm. Conclusion The method and software developed in this study provide a simple tool for effective investigation of the behavior of Siemens linac components with gantry rotation. Such a comprehensive study has been performed for the first time on Siemens machines. PMID:26604840

  15. An automation of design and modelling tasks in NX Siemens environment with original software - generator module

    NASA Astrophysics Data System (ADS)

    Zbiciak, M.; Grabowik, C.; Janik, W.

    2015-11-01

    Nowadays the design constructional process is almost exclusively aided with CAD/CAE/CAM systems. It is evaluated that nearly 80% of design activities have a routine nature. These design routine tasks are highly susceptible to automation. Design automation is usually made with API tools which allow building original software responsible for adding different engineering activities. In this paper the original software worked out in order to automate engineering tasks at the stage of a product geometrical shape design is presented. The elaborated software works exclusively in NX Siemens CAD/CAM/CAE environment and was prepared in Microsoft Visual Studio with application of the .NET technology and NX SNAP library. The software functionality allows designing and modelling of spur and helicoidal involute gears. Moreover, it is possible to estimate relative manufacturing costs. With the Generator module it is possible to design and model both standard and non-standard gear wheels. The main advantage of the model generated in such a way is its better representation of an involute curve in comparison to those which are drawn in specialized standard CAD systems tools. It comes from fact that usually in CAD systems an involute curve is drawn by 3 points that respond to points located on the addendum circle, the reference diameter of a gear and the base circle respectively. In the Generator module the involute curve is drawn by 11 involute points which are located on and upper the base and the addendum circles therefore 3D gear wheels models are highly accurate. Application of the Generator module makes the modelling process very rapid so that the gear wheel modelling time is reduced to several seconds. During the conducted research the analysis of differences between standard 3 points and 11 points involutes was made. The results and conclusions drawn upon analysis are shown in details.

  16. Secondary neutron spectra from modern Varian, Siemens, and Elekta linacs with multileaf collimators

    PubMed Central

    Howell, Rebecca M.; Kry, Stephen F.; Burgett, Eric; Hertel, Nolan E.; Followill, David S.

    2009-01-01

    Neutrons are a by-product of high-energy x-ray radiation therapy (threshold for [γ,n] reactions in high-Z material ∼7 MeV). Neutron production varies depending on photon beam energy as well as on the manufacturer of the accelerator. Neutron production from modern linear accelerators (linacs) has not been extensively compared, particularly in terms of the differences in the strategies that various manufacturers have used to implement multileaf collimators (MLCs) into their linac designs. However, such information is necessary to determine neutron dose equivalents for different linacs and to calculate vault shielding requirements. The purpose of the current study, therefore, was to measure the neutron spectra from the most up-to-date linacs from three manufacturers: Varian 21EX operating at 15, 18, and 20 MV, Siemens ONCOR operating at 15 and 18 MV, and Elekta Precise operating at 15 and 18 MV. Neutron production was measured by means of gold foil activation in Bonner spheres. Based on the measurements, the authors determined neutron spectra and calculated the average energy, total neutron fluence, ambient dose equivalent, and neutron source strength. The shapes of the neutron spectra did not change significantly between accelerators or even as a function of treatment energy. However, the neutron fluence, and therefore the ambient dose equivalent, did vary, increasing with increasing treatment energy. For a given nominal treatment energy, these values were always highest for the Varian linac. The current study thus offers medical physicists extensive information about the neutron production of MLC-equipped linacs currently in operation and provides them information vital for accurate comparison and prediction of neutron dose equivalents and calculation of vault shielding requirements. PMID:19810475

  17. Monte Carlo Simulation of Siemens ONCOR Linear Accelerator with BEAMnrc and DOSXYZnrc Code

    PubMed Central

    Jabbari, Keyvan; Anvar, Hossein Saberi; Tavakoli, Mohammad Bagher; Amouheidari, Alireza

    2013-01-01

    The Monte Carlo method is the most accurate method for simulation of radiation therapy equipment. The linear accelerators (linac) are currently the most widely used machines in radiation therapy centers. In this work, a Monte Carlo modeling of the Siemens ONCOR linear accelerator in 6 MV and 18 MV beams was performed. The results of simulation were validated by measurements in water by ionization chamber and extended dose range (EDR2) film in solid water. The linac's X-ray particular are so sensitive to the properties of primary electron beam. Square field size of 10 cm × 10 cm produced by the jaws was compared with ionization chamber and film measurements. Head simulation was performed with BEAMnrc and dose calculation with DOSXYZnrc for film measurements and 3ddose file produced by DOSXYZnrc analyzed used homemade MATLAB program. At 6 MV, the agreement between dose calculated by Monte Carlo modeling and direct measurement was obtained to the least restrictive of 1%, even in the build-up region. At 18 MV, the agreement was obtained 1%, except for in the build-up region. In the build-up region, the difference was 1% at 6 MV and 2% at 18 MV. The mean difference between measurements and Monte Carlo simulation is very small in both of ONCOR X-ray energy. The results are highly accurate and can be used for many applications such as patient dose calculation in treatment planning and in studies that model this linac with small field size like intensity-modulated radiation therapy technique. PMID:24672765

  18. 26 CFR 1.6042-3T - Dividends subject to reporting (temporary).

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 26 Internal Revenue 13 2014-04-01 2014-04-01 false Dividends subject to reporting (temporary). 1.6042-3T Section 1.6042-3T Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY... paragraph (b)(1)(iii) may be reportable, for purposes of chapter 3 of the Internal Revenue Code...

  19. 26 CFR 1.702-3T - 4-Year spread (temporary).

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 26 Internal Revenue 8 2010-04-01 2010-04-01 false 4-Year spread (temporary). 1.702-3T Section 1... (CONTINUED) INCOME TAXES Partners and Partnerships § 1.702-3T 4-Year spread (temporary). (a) Applicability... spread. A partner may elect out of the rules of paragraph (b) of this section by meeting the...

  20. 26 CFR 1.702-3T - 4-Year spread (temporary).

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 26 Internal Revenue 8 2012-04-01 2012-04-01 false 4-Year spread (temporary). 1.702-3T Section 1... (CONTINUED) INCOME TAXES (CONTINUED) Partners and Partnerships § 1.702-3T 4-Year spread (temporary). (a... taxable year for the first taxable year beginning after December 31, 1986 (partnership's year of...

  1. Active form Notch4 promotes the proliferation and differentiation of 3T3-L1 preadipocytes

    SciTech Connect

    Lai, Peng-Yeh; Tsai, Chong-Bin; Department of Ophthalmology, Chiayi Christian Hospital, Chiayi 600, Taiwan, ROC ; Tseng, Min-Jen

    2013-01-18

    Highlights: ► Notch4IC modulates the ERK pathway and cell cycle to promote 3T3-L1 proliferation. ► Notch4IC facilitates 3T3-L1 differentiation by up-regulating proadipogenic genes. ► Notch4IC promotes proliferation during the early stage of 3T3-L1 adipogenesis. ► Notch4IC enhances differentiation during subsequent stages of 3T3-L1 adipogenesis. -- Abstract: Adipose tissue is composed of adipocytes, which differentiate from precursor cells in a process called adipogenesis. Many signal molecules are involved in the transcriptional control of adipogenesis, including the Notch pathway. Previous adipogenic studies of Notch have focused on Notch1 and HES1; however, the role of other Notch receptors in adipogenesis remains unclear. Q-RT-PCR analyses showed that the augmentation of Notch4 expression during the differentiation of 3T3-L1 preadipocytes was comparable to that of Notch1. To elucidate the role of Notch4 in adipogenesis, the human active form Notch4 (N4IC) was transiently transfected into 3T3-L1 cells. The expression of HES1, Hey1, C/EBPδ and PPARγ was up-regulated, and the expression of Pref-1, an adipogenic inhibitor, was down-regulated. To further characterize the effect of N4IC in adipogenesis, stable cells expressing human N4IC were established. The expression of N4IC promoted proliferation and enhanced differentiation of 3T3-L1 cells compared with those of control cells. These data suggest that N4IC promoted proliferation through modulating the ERK pathway and the cell cycle during the early stage of 3T3-L1 adipogenesis and facilitated differentiation through up-regulating adipogenic genes such as C/EBPα, PPARγ, aP2, LPL and HSL during the middle and late stages of 3T3-L1 adipogenesis.

  2. Comparison of reconstruction methods and quantitative accuracy in Siemens Inveon PET scanner

    NASA Astrophysics Data System (ADS)

    Ram Yu, A.; Kim, Jin Su; Kang, Joo Hyun; Moo Lim, Sang

    2015-04-01

    PET reconstruction is key to the quantification of PET data. To our knowledge, no comparative study of reconstruction methods has been performed to date. In this study, we compared reconstruction methods with various filters in terms of their spatial resolution, non-uniformities (NU), recovery coefficients (RCs), and spillover ratios (SORs). In addition, the linearity of reconstructed radioactivity between linearity of measured and true concentrations were also assessed. A Siemens Inveon PET scanner was used in this study. Spatial resolution was measured with NEMA standard by using a 1 mm3 sized 18F point source. Image quality was assessed in terms of NU, RC and SOR. To measure the effect of reconstruction algorithms and filters, data was reconstructed using FBP, 3D reprojection algorithm (3DRP), ordered subset expectation maximization 2D (OSEM 2D), and maximum a posteriori (MAP) with various filters or smoothing factors (β). To assess the linearity of reconstructed radioactivity, image quality phantom filled with 18F was used using FBP, OSEM and MAP (β =1.5 & 5 × 10-5). The highest achievable volumetric resolution was 2.31 mm3 and the highest RCs were obtained when OSEM 2D was used. SOR was 4.87% for air and 3.97% for water, obtained OSEM 2D reconstruction was used. The measured radioactivity of reconstruction image was proportional to the injected one for radioactivity below 16 MBq/ml when FBP or OSEM 2D reconstruction methods were used. By contrast, when the MAP reconstruction method was used, activity of reconstruction image increased proportionally, regardless of the amount of injected radioactivity. When OSEM 2D or FBP were used, the measured radioactivity concentration was reduced by 53% compared with true injected radioactivity for radioactivity <16 MBq/ml. The OSEM 2D reconstruction method provides the highest achievable volumetric resolution and highest RC among all the tested methods and yields a linear relation between the measured and true

  3. A method of computer aided design with self-generative models in NX Siemens environment

    NASA Astrophysics Data System (ADS)

    Grabowik, C.; Kalinowski, K.; Kempa, W.; Paprocka, I.

    2015-11-01

    Currently in CAD/CAE/CAM systems it is possible to create 3D design virtual models which are able to capture certain amount of knowledge. These models are especially useful in an automation of routine design tasks. These models are known as self-generative or auto generative and they can behave in an intelligent way. The main difference between the auto generative and fully parametric models consists in the auto generative models ability to self-organizing. In this case design model self-organizing means that aside from the possibility of making of automatic changes of model quantitative features these models possess knowledge how these changes should be made. Moreover they are able to change quality features according to specific knowledge. In spite of undoubted good points of self-generative models they are not so often used in design constructional process which is mainly caused by usually great complexity of these models. This complexity makes the process of self-generative time and labour consuming. It also needs a quite great investment outlays. The creation process of self-generative model consists of the three stages it is knowledge and information acquisition, model type selection and model implementation. In this paper methods of the computer aided design with self-generative models in NX Siemens CAD/CAE/CAM software are presented. There are the five methods of self-generative models preparation in NX with: parametric relations model, part families, GRIP language application, knowledge fusion and OPEN API mechanism. In the paper examples of each type of the self-generative model are presented. These methods make the constructional design process much faster. It is suggested to prepare this kind of self-generative models when there is a need of design variants creation. The conducted research on assessing the usefulness of elaborated models showed that they are highly recommended in case of routine tasks automation. But it is still difficult to distinguish

  4. Ketoprofen S(+) enantiomer inhibits prostaglandin production and cell growth in 3T6 fibroblast cultures.

    PubMed

    Sánchez, T; Moreno, J J

    1999-04-01

    The ketoprofen S(+) enantiomer inhibits with great stereoselectivity both prostaglandin H synthase isoenzymes. Thus, the biological effects of ketoprofen on inflammation are due almost entirely to the S(+) isomer. Here, we report that the S(+) enantiomer, at doses that inhibit prostaglandin synthesis, is effective in reducing DNA synthesis and 3T6 fibroblast growth. Our data suggest that prostaglandins are involved in the control of 3T6 fibroblast growth and that the effect of the ketoprofen S(+) enantiomer on 3T6 proliferation is correlated with its effects on prostaglandin H synthase and prostaglandin production. PMID:10323281

  5. Optimization of Coronary Whole-Heart MRA Free Breathing Technique at 3T

    PubMed Central

    Gharib, Ahmed M.; Abd-Elmoniem, Khaled Z.; Herzka, Daniel A.; Ho, Vincent B.; Locklin, Julie; Tzatha, Efstathia; Stuber, Matthias; Pettigrew, Roderic I

    2011-01-01

    Four different techniques for 3T whole-heart coronary MRA using free-breathing 3D segmented parallel imaging and adiabatic T2-Prep were assessed. Coronary MRA at 3T is improved by shortening the acquisition window more than employing the highest spatial resolution. Double oblique whole-heart acquisitions result in better overall image quality and allow for better delineation of the LAD. It is possible to attain shorter acquisition windows and a smaller voxel size at 3T than previously reported at 1.5T. PMID:21871751

  6. On (2,3,t)-generations for the Conway group Co1

    NASA Astrophysics Data System (ADS)

    Ali, Faryad

    2013-09-01

    It is well known that all sporadic simple groups are (2,3,t)-generated, with the exception of M11, M22, M23 and McL. A group G is said to be (2,3,t)-generated if it can be generated by an involution x and an element y of order 3 such that o(xy) = t. In the present article, we investigate all (2,3,t)-generations for the Conway's first sporadic simple group Co1, where t is any odd divisor of |Co1|. Computations are carried out with the aid of computer algebra system GAP-Groups, Algorithms and Programming.

  7. Application of the Trio-Tri-Star Carpal Wrist for use in a Solar Array Tracking Mechanism for the Momentum-eXchange/Electrodynamic Reboost (MXER) Tether Concept

    NASA Technical Reports Server (NTRS)

    Robertson, Jeffrey

    2006-01-01

    This paper describes the application of the Trio-Tri-Star Carpal Wrist to the Momentum Exchange Electro-Dynamic Re-boost (MXER) tether, an advanced space transportation concept being developed by the In-Space Propulsion Technology Office at NASA s Marshall Space Flight Center in Huntsville, Alabama. Dr. Joseph Bonometti and Mr. Kirk Sorensen are the principal engineers. In the paper, a brief overview of the MXER concept is given, with an emphasis on the design problem that this wrist is designed to address. The Trio-Tri-Star Carpal Wrist, a three degree of freedom parallel manipulator, invented by Dr. Stephen J. Canfield of Tennessee Tech University, is described with an overview of wrist geometry, kinematics, and stress analysis. A working model of the wrist was assembled at MSFC using Dr. Canfield s prototype to demonstrate its operation. Finally, a design description and supporting analysis of a MXER flight concept wrist is given, with recommendations for future development work.

  8. Mutational analysis of 33 autosomal short tandem repeat (STR) loci in southwest Chinese Han population based on trio parentage testing.

    PubMed

    Jin, Bo; Su, Qin; Luo, Haibo; Li, Yingbi; Wu, Jin; Yan, Jing; Hou, Yiping; Liang, Weibo; Zhang, Lin

    2016-07-01

    Mutation rates and 95% CI of 33 short tandem repeat (STR) loci (D1S2142, D2S1338, D2S441, D3S1358, D3S1754, D5S818, D6S1043, D7S3048, D7S820, D8S1132, D8S1179, D10S1248, D11S2368, D12S391, D13S1492, D13S317, D13S325, D14S306, D15S659, D16S539, D18S1364, D18S51, D19S433, D20S161, D21S11, D22GATA198B05, CSF1PO, FGA, Penta D, Penta E, TH01, TPOX, and vWA) were investigated through more than 424,000 parent-child meiotic transfers obtained from 10636 trios parentage testing cases in southwest Chinese Han population. Overall, 297, including 292 single-step, 4 double-step and 1 triple-step mutation events were observed. The average mutation rate was 0.70×10(-3). Most of the locus-specific mutation rates (varied from 0.20×10(-3) to 1.96×10(-3)) were lower than the other datasets (p<0.05). Mutations of 7 loci are reported for the first time. Mutation rates varied with population from different ethnicities and geographical regions. There was no significant difference between mutation expansion and contraction (∼1.04:1). Paternal origin mutations occurred more frequently than maternal origin ones (∼5.02:1). In addition, mutation rates indicated positive correlation with the expected heterozygosity (He) and geometric mean of longest run of perfect repeats (LRPR), respectively. Short alleles showed a trend toward mutation gain while long alleles trended toward mutation loss. A credible forensic dataset for locus-specific mutation rates of 33 loci has been established based upon strict inclusion criteria of large-sized parents/child-trio cases. PMID:27045978

  9. Correlation analysis between S3T and SFT/MTK vector magnetograms

    NASA Astrophysics Data System (ADS)

    Liang, H. F.; Sin, S. A.; Ma, L.

    2008-07-01

    Three components of the vector magnetic field, flux density B, inclination γ and azimuth χ of the active region NOAA10507 are derived from the two-dimensional Stokes spectral data obtained by the S3T at the Yunnan Observatory. The distributions of the longitudinal magnetic field and the transverse magnetic field are contoured on the basis of the three components. The distributions indicate that the active region is a very complicated sunspot group which is mainly composed of five sunspots, including one of negative polarity and four positive ones. Comparing the vector magnetograms obtained by S3T and the SFT/MTK, it is found that there is basic agreement on the longitudinal fields of S3T and SFT/MTK magnetograms with a correlation coefficient ρ=0.842, and the two distributions of transverse magnetic field obtained by S3T and SFT/MTK have correlation coefficients, ρ=0.423 and ρ=0.72.

  10. Regulatory T Cells in Melanoma Revisited by a Computational Clustering of FOXP3+ T Cell Subpopulations

    PubMed Central

    Fujii, Hiroko; Josse, Julie; Tanioka, Miki; Miyachi, Yoshiki; Husson, François

    2016-01-01

    CD4+ T cells that express the transcription factor FOXP3 (FOXP3+ T cells) are commonly regarded as immunosuppressive regulatory T cells (Tregs). FOXP3+ T cells are reported to be increased in tumor-bearing patients or animals and are considered to suppress antitumor immunity, but the evidence is often contradictory. In addition, accumulating evidence indicates that FOXP3 is induced by antigenic stimulation and that some non-Treg FOXP3+ T cells, especially memory-phenotype FOXP3low cells, produce proinflammatory cytokines. Accordingly, the subclassification of FOXP3+ T cells is fundamental for revealing the significance of FOXP3+ T cells in tumor immunity, but the arbitrariness and complexity of manual gating have complicated the issue. In this article, we report a computational method to automatically identify and classify FOXP3+ T cells into subsets using clustering algorithms. By analyzing flow cytometric data of melanoma patients, the proposed method showed that the FOXP3+ subpopulation that had relatively high FOXP3, CD45RO, and CD25 expressions was increased in melanoma patients, whereas manual gating did not produce significant results on the FOXP3+ subpopulations. Interestingly, the computationally identified FOXP3+ subpopulation included not only classical FOXP3high Tregs, but also memory-phenotype FOXP3low cells by manual gating. Furthermore, the proposed method successfully analyzed an independent data set, showing that the same FOXP3+ subpopulation was increased in melanoma patients, validating the method. Collectively, the proposed method successfully captured an important feature of melanoma without relying on the existing criteria of FOXP3+ T cells, revealing a hidden association between the T cell profile and melanoma, and providing new insights into FOXP3+ T cells and Tregs. PMID:26864030

  11. In vivo CEST Imaging of Creatine (CrCEST) in Skeletal Muscle at 3T

    PubMed Central

    Kogan, Feliks; Haris, Mohammad; Debrosse, Catherine; Singh, Anup; Nanga, Ravi P.; Cai, Kejia; Hariharan, Hari; Reddy, Ravinder

    2013-01-01

    Purpose To characterize the chemical exchange saturation transfer (CEST) based technique to measure free creatine (Cr), a key component of muscle energy metabolism, distribution in skeletal muscle with high spatial resolution before and after exercise at 3T. Methods CrCEST saturation parameters were empirically optimized for 3T. CEST, T2, magnetization transfer ratio (MTR) and 31P magnetic resonance spectroscopy (MRS) acquisitions of the lower leg were performed before and after mild plantar flexion exercise on a 3T whole-body MR scanner on 6 healthy volunteers. Results The feasibility of imaging Cr changes in skeletal muscle following plantar flexion exercise using CrCEST was demonstrated at 3T. This technique exhibited good spatial resolution and was able to differentiate differences in muscle utilization among subjects. CrCEST results were compared with 31P MRS results showing good agreement in the Cr and PCr recovery kinetics. A relationship of 0.45 % CrCESTasym/mM Cr was observed across all subjects. Conclusion Demonstrated the CrCEST technique could be applied at 3T to measure dynamic changes in creatine in in vivo muscle. The widespread availability and clinical applicability of 3T scanners has the potential to clinically advance this method. PMID:24925857

  12. [Envelope protein of Jaagsiekte sheep retrovious expressed in NIH3T3 cells promotes cell proliferation].

    PubMed

    DU, Fangyuan; Chen, Dayong; Zhang, Yufei; Sun, Xiaolin; Guo, Wenqing; Liu, Shuying

    2016-09-01

    Objective To explore the influence of the exogenous Jaagsiekte sheep retrovious (exJSRV) envelope protein (Env) on NIH3T3 cell proliferation. Methods A recombinant plasmid pcDNA4/myc-His/exJSRV- env carrying exJSRV- env gene was constructed, and then the correctness of the recombinant plasmid was identified by PCR, restriction enzyme digestion and sequencing. The recombinant plasmid pcDNA4/myc-His/exJSRV- env was transiently transfected into NIH3T3 cells by Lipofectamine(TM) LTX. After the transfection of the recombinant plasmid, the expression of exJSRV- env was detected by reverse transcription PCR and Western blotting. The effect of Env on cell proliferation was investigated by CCK-8 assay and plate colony formation assay. Results The recombinant eukaryotic expression plasmid containing exJSRV- env was successfully constructed as identified by PCR, restriction enzyme identification and sequencing. After the recombinant plasmid was transiently transfected into NIH3T3 cells, reverse transcription PCR and Western blotting showed the expression of exJSRV- env , and Env promoted NIH3T3 cell proliferation significantly. Conclusion JSRV Env was expressed successfully in the NIH3T3 cells and promoted the proliferation of NIH3T3 cells. PMID:27609573

  13. Analysis of case-parent trios at a locus with a deletion allele: association of GSTM1 with autism

    PubMed Central

    Buyske, Steven; Williams, Tanishia A; Mars, Audrey E; Stenroos, Edward S; Ming, Sue X; Wang, Rong; Sreenath, Madhura; Factura, Marivic F; Reddy, Chitra; Lambert, George H; Johnson, William G

    2006-01-01

    Background Certain loci on the human genome, such as glutathione S-transferase M1 (GSTM1), do not permit heterozygotes to be reliably determined by commonly used methods. Association of such a locus with a disease is therefore generally tested with a case-control design. When subjects have already been ascertained in a case-parent design however, the question arises as to whether the data can still be used to test disease association at such a locus. Results A likelihood ratio test was constructed that can be used with a case-parents design but has somewhat less power than a Pearson's chi-squared test that uses a case-control design. The test is illustrated on a novel dataset showing a genotype relative risk near 2 for the homozygous GSTM1 deletion genotype and autism. Conclusion Although the case-control design will remain the mainstay for a locus with a deletion, the likelihood ratio test will be useful for such a locus analyzed as part of a larger case-parent study design. The likelihood ratio test has the advantage that it can incorporate complete and incomplete case-parent trios as well as independent cases and controls. Both analyses support (p = 0.046 for the proposed test, p = 0.028 for the case-control analysis) an association of the homozygous GSTM1 deletion genotype with autism. PMID:16472391

  14. WES in a family trio suggests involvement of TECPR2 in a complex form of progressive motor neuron disease.

    PubMed

    Covone, A E; Fiorillo, C; Acquaviva, M; Trucco, F; Morana, G; Ravazzolo, R; Minetti, C

    2016-08-01

    We have performed whole-exome sequencing in a family trio with a 16-year-old girl suffering of progressive motor neuron disease. There was no family history of the disease and no parental consanguinity. Our exome analysis indicated the proband as a compound heterozygote for two missense variants in the TECPR2 gene according to a recessive mode of inheritance. The TECPR2 gene has been reported as a positive regulator of autophagy which is an essential mechanism for maintaining neuron homeostasis and survival and plays a key role in major adult and pediatric neurodegenerative diseases. Variants in this gene have been found responsible for a recently described form of hereditary spastic paraplegia called SPG49 in two previous reports. We propose that both variants causing amino acid substitution, p.Leu684Val and p.Thr903Met, inherited in trans-phase compound heterozygote form, can be responsible for the phenotype observed in our patient. We also consider the possible contribution of a heterozygous variant in the SPG7 gene. Sanger sequencing confirmed the segregation of variants within the family tree including the patient's unaffected brother. PMID:27406698

  15. Temperature induced modulation of lipid oxidation and lipid accumulation in palmitate-mediated 3T3-L1 adipocytes and 3T3-L1 adipocytes.

    PubMed

    Lin, Xiaofen; Li, Yi; Leung, Polly Hangmei; Li, Jiashen; Hu, Junyan; Liu, Xuan; Li, Zhi

    2016-05-01

    Human skin temperature can vary widely depending on anatomical location and ambient temperature. It is also known that local changes in skin and subcutaneous temperature can affect fat metabolism. This study aimed to explore the potential effects of surrounding thermal environment on fat by investigating cell viability, lipid oxidation, and lipid accumulation in 3T3-L1 adipocytes and palmitate-treated adipocytes after 4h incubation. No significant differences of viability in 3T3-L1 adipocytes were detected under different temperature conditions. Despite no significant increase being observed under warm temperature (39°C) conditions, a similarly significant suppression of intracellular reactive oxygen species (ROS) and lipid peroxidation were found in 3T3-L1 adipocytes and palmitate-treated adipocytes under 4h exposure to cooler temperatures of 31-33°C (P<0.01). ROS, chemically reactive molecules containing oxygen, are currently understood to be a major contributor to oxidantive stress in obesity. Additionally, cooler temperatures (31-33°C) could improve the size of lipid droplets in 3T3-L1 adipocytes (P<0.01), but no significant effect was generated by temperature change on lipid droplets in palmitate-treated adipocytes. In the palmitate-induced adiposity model, although excessive ROS and lipid peroxidation has been attenuated by temperature decrease (P<0.01), it still does not positively modulate lipid droplet size (P>0.05) and remedy the palmitate damage induced cell death (P<0.01). These findings provide preliminary support for potential interventions based on temperature manipulation for cell metabolism of adipocytes. PMID:27157327

  16. Prolonged Induction Activates Cebpα Independent Adipogenesis in NIH/3T3 Cells

    PubMed Central

    Shao, Hsiao-Yun; Hsu, Hsue-Yin; Wu, Kuan-Sju; Hee, Siow-Wey; Chuang, Lee-Ming; Yeh, Jih-I

    2013-01-01

    Background 3T3-L1 cells are widely used to study adipogenesis and insulin response. Their adipogenic potential decreases with time in the culture. Expressing exogenous genes in 3T3-L1 cells can be challenging. This work tries to establish and characterize an alternative model of cultured adipocytes that is easier to work with than the 3T3-L1 cells. Methodology/Principal Findings Induced cells were identified as adipocytes based on the following three characteristics: (1) Accumulation of triglyceride droplets as demonstrated by oil red O stain. (2) Transport rate of 2-deoxyglucose increased after insulin stimulation. (3) Expression of fat specific genes such as Fabp4 (aP2), Slc2a4 (Glut4) and Pparg (PPARγ). Among the cell lines induced under different conditions in this study, only NIH/3T3 cells differentiated into adipocytes after prolonged incubation in 3T3-L1 induction medium containing 20% instead of 10% fetal bovine serum. Rosiglitazone added to the induction medium shortened the incubation period from 14 to 7 days. The PI3K/AKT pathway showed similar changes upon insulin stimulation in these two adipocytes. C/EBPα mRNA was barely detectable in NIH/3T3 adipocytes. NIH/3T3 adipocytes induced in the presence of rosiglitazone showed higher 2-deoxyglucose transport rate after insulin stimulation, expressed less Agt (angiotensinogen) and more PPARγ. Knockdown of C/EBPα using shRNA blocked 3T3-L1 but not NIH/3T3 cell differentiation. Mouse adipose tissues from various anatomical locations showed comparable levels of C/EBPα mRNA. Conclusions/Significance NIH/3T3 cells were capable of differentiating into adipocytes without genetic engineering. They were an adipocyte model that did not require the reciprocal activation between C/EBPα and PPARγ to differentiate. Future studies in the C/EBPα independent pathways leading to insulin responsiveness may reveal new targets to diabetes treatment. PMID:23326314

  17. Alterations in insulin binding accompanying differentiation of 3T3-L1 preadipocytes.

    PubMed Central

    Reed, B C; Kaufmann, S H; Mackall, J C; Student, A K; Lane, M D

    1977-01-01

    Expression of the adipocyte phenotype by differentiating 3T3-L1 preadipocytes occurs upon exposure of the cells to insulin. Differentiation-linked changes in 125I-labeled insulin binding to 3T3-L1 cells were monitored and compared with those in nondifferentiating 3T3-C2 controls treated similarly. Without chronic insulin treatment, 3T3-L1 cells failed to express the adipocyte phenotype but maintained a level of 25,000-35,000 insulin-binding sites per cell. Treatment of 3T3-L1 cells with insulin resulted in an initial suppression of insulin binding followed by a 12-fold increase that paralleled the appearance of differentiated cells. A maximum of 170,000 insulin-binding sites per cell was attained for a population in which greater than 75% of the cells had differentiated. The increase of insulin receptor level appears to be differentiation-dependent and is not a general response of cells to the culture conditions. 3T3-C2 cells maintained in the presence of insulin for 30 days exhibited the undifferentiated phenotype and suppressed levels of insulin binding (35,000 sites per cell). The binding capacity of 3T3-L1 cells for epidermal growth factor remained unchanged between 25,000 and 40;000 sites per cell and was independent of the state of differentiation. Thus, induction by insulin in receptor-specific changes. Insulin receptors increase in number but epidermal growth factor receptors remain constant. PMID:303773

  18. The effect of myostatin on proliferation and lipid accumulation in 3T3-L1 preadipocytes.

    PubMed

    Zhu, Hui Juan; Pan, Hui; Zhang, Xu Zhe; Li, Nai Shi; Wang, Lin Jie; Yang, Hong Bo; Gong, Feng Ying

    2015-06-01

    Myostatin is a critical negative regulator of skeletal muscle development, and has been reported to be involved in the progression of obesity and diabetes. In the present study, we explored the effects of myostatin on the proliferation and differentiation of 3T3-L1 preadipocytes by using 3-[4,5-dimethylthiazol-2-yl] 2,5-diphenyl tetrazolium bromide spectrophotometry, intracellular triglyceride (TG) assays, and real-time quantitative RT-PCR methods. The results indicated that recombinant myostatin significantly promoted the proliferation of 3T3-L1 preadipocytes and the expression of proliferation-related genes, including Cyclin B2, Cyclin D1, Cyclin E1, Pcna, and c-Myc, and IGF1 levels in the medium of 3T3-L1 were notably upregulated by 35.2, 30.5, 20.5, 33.4, 51.2, and 179% respectively (all P<0.01) in myostatin-treated 3T3-L1 cells. Meanwhile, the intracellular lipid content of myostatin-treated cells was notably reduced as compared with the non-treated cells. Additionally, the mRNA levels of Pparγ, Cebpα, Gpdh, Dgat, Acs1, Atgl, and Hsl were significantly downregulated by 22-76% in fully differentiated myostatin-treated adipocytes. Finally, myostatin regulated the mRNA levels and secretion of adipokines, including Adiponectin, Resistin, Visfatin, and plasminogen activator inhibitor-1 (PAI-1) in 3T3-L1 adipocytes (all P<0.001). Above all, myostatin promoted 3T3-L1 proliferation by increasing the expression of cell-proliferation-related genes and by stimulating IGF1 secretion. Myostatin inhibited 3T3-L1 adipocyte differentiation by suppressing Pparγ and Cebpα expression, which consequently deceased lipid accumulation in 3T3-L1 cells by inhibiting the expression of critical lipogenic enzymes and by promoting the expression of lipolytic enzymes. Finally, myostatin modulated the expression and secretion of adipokines in fully differentiated 3T3-L1 adipocytes. PMID:25878062

  19. Protein turnover and cellular autophagy in growing and growth-inhibited 3T3 cells

    SciTech Connect

    Papadopoulos, T.; Pfeifer, U. )

    1987-07-01

    The relationship between growth, protein degradation, and cellular autophagy was tested in growing and in growth-inhibited 3T3 cell monolayers. For the biochemical evaluation of DNA and protein metabolism, growth-inhibited 3T3 cell monolayers with high cell density and growing 3T3 cell monolayers with low cell density were labeled simultaneously with ({sup 14}C)thymidine and ({sup 3}H)leucine. The evaluation of the DNA turnover and additional ({sup 3}H)thymidine autoradiography showed that 24 to 5% of 3T3 cells continue to replicate even in the growth-inhibited state, where no accumulation of protein and DNA can be observed. Cell loss, therefore, has to be assumed to compensate for the ongoing cell proliferation. When the data of protein turnover were corrected for cell loss, it was found that the rate constant of protein synthesis in nongrowing monolayers was reduced to half the value found in growing monolayers. Simultaneously, the rate constant of protein degradation in nongrowing monolayers was increased to about 1.5-fold the value of growing monolayers. These data are in agreement with the assumption that cellular autophagy represents a major pathway of regulating protein degradation in 3T3 cells and that the regulation of autophagic protein degradation is of relevance for the transition from a growing to a nongrowing state.

  20. Epac, not PKA catalytic subunit, is required for 3T3-L1 preadipocyte differentiation.

    PubMed

    Ji, Zhenyu; Mei, Fang C; Cheng, Xiaodong

    2010-01-01

    Cyclic AMP plays a critical role in adipocyte differentiation and maturation. However, it is not clear which of the two intracellular cAMP receptors, exchange protein directly activated by cAMP/cAMP-regulated guanine nucleotide exchange factor or protein kinase A/cAMP-dependent protein kinase, is essential for cAMP-mediated adipocyte differentiation. In this study, we utilized a well-defined adipose differentiation model system, the murine preadipocyte line 3T3-L1, to address this issue. We showed that knocking down Epac expression in 3T3-L1 cells using lentiviral based small hairpin RNAs down-regulated peroxisome proliferator-activated receptor gamma expression and dramatically inhibited adipogenic conversion of 3T3-L1 cells while inhibiting PKA catalytic subunit activity by two mechanistically distinct inhibitors, heat stable protein kinase inhibitor and H89, had no effect on 3T3-L1 adipocyte differentiation. Moreover, cAMP analog selectively activating Epac was not able to stimulate adipogenic conversion. Our study demonstrated that while PKA catalytic activity is dispensable, activation of Epac is necessary but not sufficient for adipogenic conversion of 3T3-L1 cells. PMID:20036887

  1. DNA Methylation Suppresses Leptin Gene in 3T3-L1 Adipocytes

    PubMed Central

    Kuroda, Masashi; Tominaga, Ayako; Nakagawa, Kasumi; Nishiguchi, Misa; Sebe, Mayu; Miyatake, Yumiko; Kitamura, Tadahiro; Tsutsumi, Rie; Harada, Nagakatsu; Nakaya, Yutaka; Sakaue, Hiroshi

    2016-01-01

    Leptin is a key regulator of energy intake and expenditure. This peptide hormone is expressed in mouse white adipose tissue, but hardly expressed in 3T3-L1 adipocytes. Using bisulfite sequencing, we found that CpG islands in the leptin promoter are highly methylated in 3T3-L1cells. 5-azacytidine, an inhibitor of DNA methyltransferase, markedly increased leptin expression as pre-adipocytes matured into adipocytes. Remarkably, leptin expression was stimulated by insulin in adipocytes derived from precursor cells exposed to 5-azacytidine, but suppressed by thiazolidinedione and dexamethasone. In contrast, adipocytes derived from untreated precursor cells were unresponsive to both 5-azacytidine and hormonal stimuli, although lipid accumulation was sufficient to boost leptin expression in the absence of demethylation. Taken together, the results suggest that leptin expression in 3T3-L1 cells requires DNA demethylation prior to adipogenesis, transcriptional activation during adipogenesis, and lipid accumulation after adipogenesis. PMID:27494408

  2. Effect of Biodegradable Shape-Memory Polymers on Proliferation of 3T3 Cells

    NASA Astrophysics Data System (ADS)

    Xu, Shuo-Gui; Zhang, Peng; Zhu, Guang-Ming; Jiang, Ying-Ming

    2011-07-01

    This article evaluates the in vitro biocompatibility for biodegradable shape-memory polymers (BSMP) invented by the authors. 3T3 cells (3T3-Swiss albino GNM 9) of primary and passaged cultures were inoculated into two kinds of carriers: the BSMP carrier and the control group carrier. Viability, proliferation, and DNA synthesis (the major biocompatibility parameters), were measured and evaluated for both the BSMP and naked carrier via cell growth curve analysis, MTT colorimetry and addition of 3H-TdR to culture media. The results showed that there was no difference between the BSMP carrier and the control dish in terms of viability, proliferation, and metabolism of the 3T3 cells. Overall, the BSMP carrier provides good biocompatibility and low toxicity to cells in vitro, and could indicate future potential for this medium as a biological material for implants in vivo.

  3. Transformation of human cells by DNAs ineffective in transformation of NIH 3T3 cells

    SciTech Connect

    Sutherland, B.M.; Bennett, P.B.; Freeman, A.G.; Moore, S.P.; Strickland, P.T.

    1985-04-01

    Neonatal human foreskin fibroblasts can be transformed to anchorage-independent growth by transfection with DNAs inefficient in transforming NIH 3T3 cells. Human cells transfected with DNA from GM 1312, a multiple myeloma cell line, or MOLT-4, a permanent lymphoblast line, grow without anchorage at a much higher frequency than do the parental cells and their DNAs can transform human cell recipients to anchorage-independent growth; they have extended but not indefinite life spans and are nontumorigenic. Human fibroblasts are also transformed by DNAs from two multiple myeloma lines that also transform 3T3 cells; however, restriction analysis suggests that different transforming genes in this DNA are acting in the human and murine systems. These results indicate that the human cell transfection system allows detection of transforming genes not effective in the 3T3 system and points out the possibility of detection of additional transforming sequences even in DNAs that do transform murine cells.

  4. 17 CFR 270.6e-3(T) - Temporary exemptions for flexible premium variable life insurance separate accounts.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 17 Commodity and Securities Exchanges 3 2013-04-01 2013-04-01 false Temporary exemptions for flexible premium variable life insurance separate accounts. 270.6e-3(T) Section 270.6e-3(T) Commodity and Securities Exchanges SECURITIES AND EXCHANGE COMMISSION (CONTINUED) RULES AND REGULATIONS, INVESTMENT COMPANY ACT OF 1940 § 270.6e-3(T)...

  5. 17 CFR 240.15b7-3T - Operational capability in a Year 2000 environment.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... Year 2000 environment. 240.15b7-3T Section 240.15b7-3T Commodity and Securities Exchanges SECURITIES... § 240.15b7-3T Operational capability in a Year 2000 environment. (a) This section applies to every... control; (ii) Have not verified your Year 2000 remediation efforts through reasonable internal testing...

  6. 17 CFR 240.15b7-3T - Operational capability in a Year 2000 environment.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... Year 2000 environment. 240.15b7-3T Section 240.15b7-3T Commodity and Securities Exchanges SECURITIES... § 240.15b7-3T Operational capability in a Year 2000 environment. (a) This section applies to every... control; (ii) Have not verified your Year 2000 remediation efforts through reasonable internal testing...

  7. 17 CFR 240.15b7-3T - Operational capability in a Year 2000 environment.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... Year 2000 environment. 240.15b7-3T Section 240.15b7-3T Commodity and Securities Exchanges SECURITIES... § 240.15b7-3T Operational capability in a Year 2000 environment. (a) This section applies to every... control; (ii) Have not verified your Year 2000 remediation efforts through reasonable internal testing...

  8. 26 CFR 1.367(a)-3T - Treatment of transfers of stock or securities to foreign corporations (temporary).

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 26 Internal Revenue 4 2013-04-01 2013-04-01 false Treatment of transfers of stock or securities to foreign corporations (temporary). 1.367(a)-3T Section 1.367(a)-3T Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY (CONTINUED) INCOME TAX (CONTINUED) INCOME TAXES (CONTINUED) Effects on Corporation § 1.367(a)-3T Treatment...

  9. 17 CFR 275.206(3)-3T - Temporary rule for principal trades with certain advisory clients.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... required by 17 CFR 240.10b-10, a conspicuous, plain English statement informing the advisory client that... trades with certain advisory clients. 275.206(3)-3T Section 275.206(3)-3T Commodity and Securities... 1940 § 275.206(3)-3T Temporary rule for principal trades with certain advisory clients. (a)...

  10. Trio engagement via plasma membrane phospholipids and the myristoyl moiety governs HIV-1 matrix binding to bilayers

    PubMed Central

    Vlach, Jiri; Saad, Jamil S.

    2013-01-01

    Localization of the HIV type-1 (HIV-1) Gag protein on the plasma membrane (PM) for virus assembly is mediated by specific interactions between the N-terminal myristoylated matrix (MA) domain and phosphatidylinositol-(4,5)-bisphosphate [PI(4,5)P2]. The PM bilayer is highly asymmetric, and this asymmetry is considered crucial in cell function. In a typical mammalian cell, the inner leaflet of the PM is enriched in phosphatidylserine (PS) and phosphatidylethanolamine (PE) and contains minor populations of phosphatidylcholine (PC) and PI(4,5)P2. There is strong evidence that efficient binding of HIV-1 Gag to membranes is sensitive not only to lipid composition and net negative charge, but also to the hydrophobic character of the acyl chains. Here, we show that PS, PE, and PC interact directly with MA via a region that is distinct from the PI(4,5)P2 binding site. Our NMR data also show that the myristoyl group is readily exposed when MA is bound to micelles or bicelles. Strikingly, our structural data reveal a unique binding mode by which the 2′-acyl chain of PS, PE, and PC lipids is buried in a hydrophobic pocket whereas the 1′-acyl chain is exposed. Sphingomyelin, a major lipid localized exclusively on the outer layer of the PM, does not bind to MA. Our findings led us to propose a trio engagement model by which HIV-1 Gag is anchored to the PM via the 1′-acyl chains of PI(4,5)P2 and PS/PE/PC and the myristoyl group, which collectively bracket a basic patch projecting toward the polar leaflet of the membrane. PMID:23401539

  11. Analytic power and sample size calculation for the genotypic transmission/disequilibrium test in case-parent trio studies

    PubMed Central

    Neumann, Christoph; Taub, Margaret A.; Younkin, Samuel G.; Beaty, Terri H.; Ruczinski, Ingo; Schwender, Holger

    2014-01-01

    Case-parent trio studies considering genotype data from children affected by a disease and from their parents are frequently used to detect single nucleotide polymorphisms (SNPs) associated with disease. The most popular statistical tests in this study design are transmission/disequlibrium tests (TDTs). Several types of these tests have been developed, e.g., procedures based on alleles or genotypes. Therefore, it is of great interest to examine which of these tests have the highest statistical power to detect SNPs associated with disease. Comparisons of the allelic and the genotypic TDT for individual SNPs have so far been conducted based on simulation studies, since the test statistic of the genotypic TDT was determined numerically. Recently, it, however, has been shown that this test statistic can be presented in closed form. In this article, we employ this analytic solution to derive equations for calculating the statistical power and the required sample size for different types of the genotypic TDT. The power of this test is then compared with the one of the corresponding score test assuming the same mode of inheritance as well as the allelic TDT based on a multiplicative mode of inheritance, which is equivalent to the score test assuming an additive mode of inheritance. This is, thus, the first time that the power of these tests are compared based on equations, yielding instant results and omitting the need for time-consuming simulation studies. This comparison reveals that the tests have almost the same power, with the score test being slightly more powerful. PMID:25123830

  12. The East Australian, Tasmantid, and Lord Howe Volcanic Chains: Possible mechanisms behind a trio of hotspot trails

    NASA Astrophysics Data System (ADS)

    Kalnins, L. M.; Cohen, B. E.; Fitton, J. G.; Mark, D. F.; Richards, F. D.; Barfod, D. N.

    2015-12-01

    The east Australian and Tasman Sea region is home to a unique example of intraplate volcanism: three long-lived, sub-parallel volcanic chains spaced only about 500 km apart. Here we present new 40Ar/39Ar results from the centre chain, the Tasmantid Seamounts, and show that the chain is strongly age-progressive, with an excellent correspondence to the age of the continental East Australian Volcanic Chain to the west and to the more limited ages available for the Lord Howe Seamount Chain to the east. Results from the Louisiade Plateau at the northern end of the Tasmantid chain suggest that it is composed of basalts of the correct age to be a large igneous province formed by the impact of the Tasmantid plume head reaching the lithosphere. This record of relative movement between the plate and the magma source over the last 55 Ma shows two clear deflections from the overall linear trend, one at 26--23 Ma, also observed in the continental chain and linked with the Ontong-Java Plateau jamming the South Melanesian subduction zone, and another at 50--43 Ma, beyond the end of the continental record and contemporaneous with the Hawaiian-Emperor bend. How does such a unique trio of volcanic chains form? The clear age progression, long lifespan, and tie to the Louisiade Plateau are classic indicators of deep-seated plumes, but it is difficult to explain how three separate plumes could remain stable for over 30 Ma when separated by little more than the radii of the plume conduits. Here we examine alternative possible explanations for this volcanic pattern, including small plumes rising from a single deep-seated plume pooling at the 660 km discontinuity, a single plume splitting around a subducting slab fragment, and small-scale convection triggered by topography on the lithosphere-asthenosphere boundary.

  13. New Clinically Feasible 3T MRI Protocol to Discriminate Internal Brain Stem Anatomy.

    PubMed

    Hoch, M J; Chung, S; Ben-Eliezer, N; Bruno, M T; Fatterpekar, G M; Shepherd, T M

    2016-06-01

    Two new 3T MR imaging contrast methods, track density imaging and echo modulation curve T2 mapping, were combined with simultaneous multisection acquisition to reveal exquisite anatomic detail at 7 canonical levels of the brain stem. Compared with conventional MR imaging contrasts, many individual brain stem tracts and nuclear groups were directly visualized for the first time at 3T. This new approach is clinically practical and feasible (total scan time = 20 minutes), allowing better brain stem anatomic localization and characterization. PMID:26869471

  14. Advances in hybrid MR-PET at 3 T and 9.4 T in humans

    NASA Astrophysics Data System (ADS)

    Jon Shah, N.; Mauler, Jörg; Neuner, Irene; Oros-Peusquens, Ana-Maria; Romanzetti, Sandro; Vahedipour, Kaveh; Felder, Jörg; Celik, Avdo; Iida, Hidehiro; Langen, Karl-Josef; Herzog, Hans

    2013-02-01

    Hybrid MR-PET data acquisition in simultaneous mode confers a number of advantages at 3 T and 9.4 T. From an MR perspective, the potential for ultra-high resolution structural imaging is discussed and example images of the cerebellum with an isotropic resolution of 320 μm are presented. Further, metabolic imaging is discussed and high-resolution images of the sodium distribution are demonstrated. Examples of tumour imaging on a 3 T MR-PET system are included and discussed.

  15. Characterization of copy number variants for CCL3L1 gene in rheumatoid arthritis for French trio families and Tunisian cases and controls.

    PubMed

    Ben Kilani, Mohamed Sahbi; Achour, Yosser; Perea, Javier; Cornelis, François; Bardin, Thomas; Chaudru, Valérie; Maalej, Abdellatif; Petit-Teixeira, Elisabeth

    2016-08-01

    Analyses of copy number variants (CNVs) for candidate genes in complex diseases are currently a promising research field. CNVs of C-C chemokine ligand 3-like 1 (CCL3L1) gene are candidate genomic factors in rheumatoid arthritis (RA). We investigated CCL3L1 CNVs association with a case-control study in Tunisians and a transmission analysis in French trio families. Relative copy number (rCN) of CCL3L1 gene was quantified by droplet digital PCR (ddPCR) in 100 French trio families (RA patients and their two parents) and in 166 RA cases and 102 healthy controls from Tunisia. We calculated odds ratio (OR) to investigate association risk for CCL3L1 CNVs in RA. rCN identified varied from 0 to 4 in the French population and from 0 to 7 in the Tunisian population. A significant difference was observed in the distribution of these rCNs between the two populations (p = 2.34 × 10(-10)), as when rCN from French and Tunisian RA patients were compared (p = 2.83 × 10(-5)). CNVs transmission in French RA trios allowed the characterization of genotypes with the presence of tandem duplication and triplication on the same chromosome. RA association tests highlighted a protective effect of rCN = 5 for CCL3L1 gene in the Tunisian population (OR = 0.056; CI 95 % [0.01-0.46]). Characterization of CCL3L1 CNVs with ddPCR methodology highlighted specific CN genotypes in a French family sample. A copy number polymorphism of a RA candidate gene was quantified, and its significant association with RA was revealed in a Tunisian sample. PMID:26728148

  16. Cdc42 and the Guanine Nucleotide Exchange Factors Ect2 and Trio Mediate Fn14-Induced Migration and Invasion of Glioblastoma Cells

    PubMed Central

    Fortin, Shannon P.; Ennis, Matthew J.; Schumacher, Cassie A.; Zylstra-Diegel, Cassandra R.; Williams, Bart O.; Ross, Julianna T.D.; Winkles, Jeffrey A.; Loftus, Joseph C.; Symons, Marc H.; Tran, Nhan L.

    2012-01-01

    Malignant glioblastomas are characterized by their ability to infiltrate into normal brain. We previously reported that binding of the multifunctional cytokine TNF-like weak inducer of apoptosis (TWEAK) to its receptor fibroblast growth factor–inducible 14 (Fn14) induces glioblastoma cell invasion via Rac1 activation. Here, we show that Cdc42 plays an essential role in Fn14-mediated activation of Rac1. TWEAK-treated glioma cells display an increased activation of Cdc42, and depletion of Cdc42 using siRNA abolishes TWEAK-induced Rac1 activation and abrogates glioma cell migration and invasion. In contrast, Rac1 depletion does not affect Cdc42 activation by Fn14, showing that Cdc42 mediates TWEAK-stimulated Rac1 activation. Furthermore, we identified two guanine nucleotide exchange factors (GEF), Ect2 and Trio, involved in TWEAK-induced activation of Cdc42 and Rac1, respectively. Depletion of Ect2 abrogates both TWEAK-induced Cdc42 and Rac1 activation, as well as subsequent TWEAK-Fn14–directed glioma cell migration and invasion. In contrast, Trio depletion inhibits TWEAK-induced Rac1 activation but not TWEAK-induced Cdc42 activation. Finally, inappropriate expression of Fn14 or Ect2 in mouse astrocytes in vivo using an RCAS vector system for glial-specific gene transfer in G-tva transgenic mice induces astrocyte migration within the brain, corroborating the in vitro importance of the TWEAK-Fn14 signaling cascade in glioblastoma invasion. Our results suggest that the TWEAK-Fn14 signaling axis stimulates glioma cell migration and invasion through two GEF-GTPase signaling units, Ect2-Cdc42 and Trio-Rac1. Components of the Fn14-Rho GEF-Rho GTPase signaling pathway present innovative drug targets for glioma therapy. PMID:22571869

  17. Viscoelastic analysis of high molecular weight, alkali-denatured DNA from mouse 3T3 cells.

    PubMed Central

    Uhlenhopp, E L

    1975-01-01

    Alkaline lysates of mouse 3T3 cells showed viscoelastic properties characteristic of very large molecules of single-stranded DNA. The viscoelastic retardation time and the sensitivity to low doses of nitrogen mustard and of X-irradiation suggest a molecular weight in excess of 10-10 daltons. Contact-inhibited cells yielded larger single strands than actively growing cells. PMID:235335

  18. 26 CFR 1.469-3T - Passive activity credit (temporary).

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 26 Internal Revenue 6 2014-04-01 2014-04-01 false Passive activity credit (temporary). 1.469-3T... activity credit (temporary). (a) Computation of passive activity credit. The taxpayer's passive activity credit for the taxable year is the amount (if any) by which— (1) The sum of all of the taxpayer's...

  19. 26 CFR 1.469-3T - Passive activity credit (temporary).

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 26 Internal Revenue 6 2013-04-01 2013-04-01 false Passive activity credit (temporary). 1.469-3T... activity credit (temporary). (a) Computation of passive activity credit. The taxpayer's passive activity credit for the taxable year is the amount (if any) by which— (1) The sum of all of the taxpayer's...

  20. 26 CFR 1.469-3T - Passive activity credit (temporary).

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 26 Internal Revenue 6 2012-04-01 2012-04-01 false Passive activity credit (temporary). 1.469-3T... activity credit (temporary). (a) Computation of passive activity credit. The taxpayer's passive activity credit for the taxable year is the amount (if any) by which— (1) The sum of all of the taxpayer's...

  1. Olanzapine induces SREBP-1-related adipogenesis in 3T3-L1 cells.

    PubMed

    Yang, Li-Hung; Chen, Tzer-Ming; Yu, Sung-Tsai; Chen, Yen-Hui

    2007-09-01

    Olanzapine is a second-generation atypical antipsychotic drug (AAPD). Major side effects of olanzapine are weight gain and development of diabetes mellitus, which are risk factors of cardiovascular diseases. The possible causes of metabolic adverse effects are known as poor satiety and increased food intake due to blockade of receptors such as 5-HT(2C) in CNS. In this study, we examine the effect of olanzapine on peripheral adipogenesis using cultured 3T3-L1 cell model. Olanzapine increased triacylglyceride (TG) accumulation during 3T3-L1 preadipocyte differentiation to mature adipocyte phenotype. TG accumulation was accompanied by overexpression of fatty acid synthase and adiponectin that are the downstream genes of sterol regulatory element binding protein-1 (SREBP-1), one of the key transcription factors in lipid homeostasis. We further consisted that mostly SREBP-1 and at a lesser extent peroxisome proliferator-activated receptor gamma (PPAR-gamma), but not CCAAT/enhancer binding protein-alpha (C/EBP-alpha), were overexpressed and activated in 3T3-L1 adipocytes exposed to olanzapine. Furthermore, we showed that olanzapine enhanced the activity of SRE-1-containing LDLR promoter in transfected 3T3-L1 adipocytes and HepG2 cells. Taken together, olanzapine may cause body weight gain not only through influencing CNS receptors, but also affecting the peripheral adipogenesis regulated by SREBP-1. PMID:17651982

  2. Anti-Obesity Effects of Starter Fermented Kimchi on 3T3-L1 Adipocytes

    PubMed Central

    Lee, Kyung-Hee; Song, Jia-Le; Park, Eui-Seong; Ju, Jaehyun; Kim, Hee-Young; Park, Kun-Young

    2015-01-01

    The anti-obesity effects of starter (Leuconostoc mesenteroides+Lactobacillus plantarum) fermented kimchi on 3T3-L1 adipocyte were studied using naturally fermented kimchi (NK), a functional kimchi (FK, NK supplemented with green tea), and FK supplemented with added starters (FKS). Oil red O staining and cellular levels of triglyceride (TG) and glycerol were used to evaluate the in vitro anti-obesity effects of these kimchis in 3T3-L1 cells. The expressions of adipogenesis/lipogenesis-related genes of peroxisome proliferator-active receptor (PPAR)-γ, CCAAT/enhance-binding protein (C/EBP)-α, and fatty acid synthase (FAS) were determined by RT-PCR. Kimchis, especially FKS, markedly decreased TG levels and increased levels of intracellular glycerol and lipid lipolysis. In addition, FKS also reduced the mRNA levels of PPAR-γ, C/EBP-α, and FAS, which are related to adipogenesis/lipogenesis in 3T3-L1 cells. These results suggest the anti-obesity effects of FKS were to due to enhanced lipolysis and reduced adipogenesis/lipogenesis in 3T3-L1 adipocytes. PMID:26770918

  3. Complete Genome Sequence of the Novel Leech Symbiont Mucinivorans hirudinis M3T

    PubMed Central

    Bomar, Lindsey

    2015-01-01

    Mucinivorans hirudinis M3T was isolated from the digestive tract of the medicinal leech, Hirudo verbana, and is the type species of a new genus within the Rikenellaceae. Here, we report the complete annotated genome sequence of this bacterium. PMID:25657285

  4. 26 CFR 1.469-3T - Passive activity credit (temporary).

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 26 Internal Revenue 6 2011-04-01 2011-04-01 false Passive activity credit (temporary). 1.469-3T... activity credit (temporary). (a) Computation of passive activity credit. The taxpayer's passive activity credit for the taxable year is the amount (if any) by which— (1) The sum of all of the taxpayer's...

  5. Effect of Gambisan on the Inhibition of Adipogenesis in 3T3-L1 Adipocytes

    PubMed Central

    Kang, Jung Won; Nam, Dongwoo; Kim, Kun Hyung; Huh, Jeong-Eun; Lee, Jae-Dong

    2013-01-01

    This study was conducted to explore the antiadipogenic effect and possible mechanism of Gambisan on 3T3-L1 cells. For quality control, Gambisan was standardized by HPLC and the standard compounds ephedrine, epigallocatechin-3-gallate, and caffeine were screened. Cultured 3T3-L1 cells that had been induced to differentiate were treated with various concentrations of Gambisan or its major component extracts (Ephedra intermedia Schrenk, Atractylodes lancea DC., and Thea sinensis L.) for 72 hours for MTT assay to determine cell viability or 10 days for LDH assay, triglyceride assay, DNA content measurement, Oil red O staining, RT-PCR, and western blot. Gambisan significantly inhibited adipogenesis in 3T3-L1 cells by reducing triglyceride contents and lipid accumulation in a dose-dependent manner without obvious cytotoxicity. Viability and DNA content in 3T3-L1 cells treated with Gambisan were significantly higher than cells treated with the major component extracts at every concentration. The anti-adipogenic effects of Gambisan appeared to be mediated by a significant downregulation of the expression of lipoprotein lipase mRNA and PPARγ, C/EBPα, and SREBP-1 protein apart from the expression of hormone-sensitive lipase. Gambisan could act as a possible therapeutic agent for obesity. However, further studies including in vivo assays and clinical trials are needed to confirm the efficacy, safety and mechanisms of the antiobesity effects of Gambisan. PMID:24069055

  6. 26 CFR 1.883-3T - Treatment of controlled foreign corporations (temporary).

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... (CONTINUED) INCOME TAX (CONTINUED) INCOME TAXES Foreign Corporations § 1.883-3T Treatment of controlled... Internal Revenue Service (see § 601.601(d)(2) of this chapter). (3) Ownership statements from... information as specified in guidance published by the Internal Revenue Service (see § 601.601(d)(2) of...

  7. 26 CFR 1.25-3T - Qualified mortgage credit certificate (Temporary).

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... applicable to underpayments of tax due to fraud (including rules relating to the statute of limitations... TAX INCOME TAXES Changes in Rates During A Taxable Year § 1.25-3T Qualified mortgage credit... the applicant's Federal tax returns for the preceding 3 years and examines each statement to...

  8. 26 CFR 1.25-3T - Qualified mortgage credit certificate (Temporary).

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... applicable to underpayments of tax due to fraud (including rules relating to the statute of limitations... TAX INCOME TAXES Changes in Rates During A Taxable Year § 1.25-3T Qualified mortgage credit... the applicant's Federal tax returns for the preceding 3 years and examines each statement to...

  9. 26 CFR 1.25-3T - Qualified mortgage credit certificate (Temporary).

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... applicable to underpayments of tax due to fraud (including rules relating to the statute of limitations... TAX INCOME TAXES Changes in Rates During A Taxable Year § 1.25-3T Qualified mortgage credit... the applicant's Federal tax returns for the preceding 3 years and examines each statement to...

  10. 26 CFR 1.25-3T - Qualified mortgage credit certificate (Temporary).

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... applicable to underpayments of tax due to fraud (including rules relating to the statute of limitations... TAX INCOME TAXES Changes in Rates During A Taxable Year § 1.25-3T Qualified mortgage credit... the applicant's Federal tax returns for the preceding 3 years and examines each statement to...

  11. 26 CFR 1.954-3T - Foreign base company sales income (temporary).

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... (CONTINUED) INCOME TAX (CONTINUED) INCOME TAXES Controlled Foreign Corporations § 1.954-3T Foreign base... 26 Internal Revenue 10 2010-04-01 2010-04-01 false Foreign base company sales income (temporary... similar establishment has substantially the same tax effect as if such branch or similar...

  12. 26 CFR 1.444-3T - Manner and time of making section 444 election (temporary).

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 26 Internal Revenue 6 2013-04-01 2013-04-01 false Manner and time of making section 444 election... (CONTINUED) INCOME TAX (CONTINUED) INCOME TAXES (CONTINUED) Accounting Periods § 1.444-3T Manner and time of... manner and at the time provided in this section. (b) Manner and time of making election—(1) General...

  13. 26 CFR 1.444-3T - Manner and time of making section 444 election (temporary).

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 26 Internal Revenue 6 2010-04-01 2010-04-01 false Manner and time of making section 444 election... (CONTINUED) INCOME TAX (CONTINUED) INCOME TAXES Accounting Periods § 1.444-3T Manner and time of making... at the time provided in this section. (b) Manner and time of making election—(1) General rule....

  14. 26 CFR 1.444-3T - Manner and time of making section 444 election (temporary).

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 26 Internal Revenue 6 2011-04-01 2011-04-01 false Manner and time of making section 444 election... (CONTINUED) INCOME TAX (CONTINUED) INCOME TAXES (CONTINUED) Accounting Periods § 1.444-3T Manner and time of... manner and at the time provided in this section. (b) Manner and time of making election—(1) General...

  15. 26 CFR 1.444-3T - Manner and time of making section 444 election (temporary).

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 26 Internal Revenue 6 2014-04-01 2014-04-01 false Manner and time of making section 444 election... (CONTINUED) INCOME TAX (CONTINUED) INCOME TAXES (CONTINUED) Accounting Periods § 1.444-3T Manner and time of... manner and at the time provided in this section. (b) Manner and time of making election—(1) General...

  16. Fluorescence lifetime imaging of lipids during 3T3-L1 cell differentiation

    NASA Astrophysics Data System (ADS)

    Song, Young Sik; Won, Young Jae; Lee, Sang-Hak; Kim, Dug Young

    2014-03-01

    Obesity is becoming a big health problem in these days. Since increased body weight is due to increased number and size of the triglyceride-storing adipocytes, many researchers are working on differentiation conditions and processes of adipocytes. Adipocytes also work as regulators of whole-body energy homeostasis by secreting several proteins that regulate processes as diverse as haemostasis, blood pressure, immune function, angiogenesis and energy balance. 3T3-L1 cells are widely used cell line for studying adipogenesis because it can differentiate into an adipocyte-like phenotype under appropriate conditions. In this paper, we propose an effective fluorescence lifetime imaging technique which can easily distinguish lipids in membrane and those in lipid droplets. Nile red dyes are attached to lipids in 3T3-L1 cells. Fluorescence lifetime images were taken for 2 week during differentiation procedure of 3T3-L1 cells into adipocytes. We used 488 nm pulsed laser with 5MHz repetition rate and emission wavelength is 520 nm of Nile Red fluorescent dye. Results clearly show that the lifetime of Nile red in lipid droplets are smaller than those in cell membrane. Our results suggest that fluorescence lifetime imaging can be a very powerful tool to monitor lipid droplet formation in adipocytes from 3T3-L1 cells.

  17. Quantitative PET imaging with the 3T MR-BrainPET

    NASA Astrophysics Data System (ADS)

    Weirich, C.; Scheins, J.; Lohmann, P.; Tellmann, L.; Byars, L.; Michel, C.; Rota Kops, E.; Brenner, D.; Herzog, H.; Shah, N. J.

    2013-02-01

    The new hybrid imaging technology of MR-PET allows for simultaneous acquisition of versatile MRI contrasts and the quantitative metabolic imaging with PET. In order to achieve the quantification of PET images with minimal residual error the application of several corrections is crucial. In this work we present our results on quantification with the 3T MR BrainPET scanner.

  18. 26 CFR 1.181-3T - Qualified film or television production (temporary).

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... with respect to the production in order to demonstrate the eligibility of the production under section... 26 Internal Revenue 3 2010-04-01 2010-04-01 false Qualified film or television production... (continued) § 1.181-3T Qualified film or television production (temporary). (a) In general. The...

  19. Antiadopogenic effects of rice hull smoke extract in 3T3-L1 cells

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The present study investigates the inhibitory effects of a rice hull smoke extract (RHSE) against adipogenesis in 3T3-L1 pre-adipocyte cells. At concentrations of 0.1% and 0.5% RHSE, MDI-induced cells were shown to reduce their cellular lipid content by about 72% and 88%, respectively, compared to ...

  20. Osteogenic gene expression of murine osteoblastic (MC3T3-E1) cells under cyclic tension

    NASA Astrophysics Data System (ADS)

    Kao, C. T.; Chen, C. C.; Cheong, U.-I.; Liu, S. L.; Huang, T. H.

    2014-08-01

    Low-level laser therapy (LLLT) can promote cell proliferation. The remodeling ability of the tension side of orthodontic teeth affects post-orthodontic stability. The purpose of the present study was to investigate the osteogenic effects of LLLT on osteoblast-like cells treated with a simulated tension system that provides a mechanical tension regimen. Murine osteoblastic (MC3T3-E1) cells were cultured in a Flexcell strain unit with programmed loads of 12% elongation at a frequency of 0.5 Hz for 24 and 48 h. The cultured cells were treated with a low-level diode laser using powers of 5 J and 10 J. The proliferation of MC3T3-E1 cells was determined using the Alamar Blue assay. The expression of osteogenic genes (type I collagen (Col-1), osteopontin (OPN), osteocalcin (OC), osteoprotegerin (OPG), receptor activator of nuclear factor kappa B ligand (RANKL), bone morphologic protein (BMP-2), and bone morphologic protein (BMP-4)) in MC3T3-E1 cells was analyzed using reverse transcription polymerase chain reaction (RT-PCR). The data were analyzed using one-way analysis of variance. The proliferation rate of tension-cultured MC3T3-E1 cells under 5 J and 10 J LLLT increased compared with that of the control group (p < 0.05). Prominent mineralization of the MC3T3-E1 cells was visible using a von Kossa stain in the 5 J LLLT group. Osteogenic genes (Col-1, OC, OPG and BMP-2) were significantly expressed in the MC3T3-E1 cells treated with 5 J and 10 J LLLT (p < 0.05). LLLT in tension-cultured MC3T3-E1 cells showed synergistic osteogenic effects, including increases in cell proliferation and Col-1, OPN, OC, OPG and BMP-2 gene expression. LLLT might be beneficial for bone remodeling on the tension side of orthodontics.

  1. Differential effect of L3T4+ cells on recovery from total-body irradiation

    SciTech Connect

    Pantel, K.; Nakeff, A. )

    1990-09-01

    We have examined the importance of L3T4+ (murine equivalent to CD4+) cells for hematopoietic regulation in vivo in unperturbed mice and mice recovering from total-body irradiation (TBI) using a cytotoxic monoclonal antibody (MoAb) raised with the GK 1.5 hybridoma. Ablating L3T4+ cells in normal (unperturbed) B6D2F1 mice substantially decreased the S-phase fraction (determined by in vivo hydroxyurea suicide) of erythroid progenitor cells (erythroid colony-forming units, CFU-E) as compared to the pretreatment level (10% +/- 14.1% (day 3 following depletion) vs 79.8% +/- 15.9%, respectively) with a corresponding decrease in the marrow content of CFU-E at this time to approximately 1% of the pretreatment value. Although the S-phase fraction of CFU-GM was decreased to 2.2% +/- 3.1% 3 days after L3T4+ cell ablation from the 21.3% +/- 8.3% pretreatment value, CFU-GM cellularity showed little change over the 3 days following anti-L3T4 treatment. Anti-L3T4 MoAb treatment had little or no effect on either the S-phase fraction or the marrow content of hematopoietic stem cells (spleen colony-forming units, CFU-S) committed to myeloerythroid differentiation. Ablating L3T4+ cells prior to a single dose of 2 Gy TBI resulted in significantly reduced marrow contents of CFU-S on day 3 and granulocyte-macrophage colony-forming units (CFU-GM) on day 6 following TBI, with little or no effect on the corresponding recovery of CFU-E. The present findings provide the first in vivo evidence that L3T4+ cells are involved in: (1) maintaining the proliferative activity of CFU-E and CFU-GM in unperturbed mice and (2) supporting the restoration of CFU-S and CFU-GM following TBI-induced myelosuppression.

  2. CLOCK promotes 3T3-L1 cell proliferation via Wnt signaling.

    PubMed

    Zhu, Zhu; Hua, Bingxuan; Xu, Lirong; Yuan, Gongsheng; Li, Ermin; Li, Xiaobo; Sun, Ning; Yan, Zuoqin; Lu, Chao; Qian, Ruizhe

    2016-07-01

    Circadian genes control most of the physiological functions including cell cycle. Cell proliferation is a critical factor in the differentiation of progenitor cells. However, the role of Clock gene in the regulation of cell cycle via wingless-type (Wnt) pathway and the relationship between Clock and adipogenesis are unclear. We found that the circadian locomotor output cycles kaput (Clock) regulated the proliferation and the adipogenesis of 3T3-L1 preadipocytes. We found that Clock attenuation inhibited the viability of 3T3-L1 preadipocytes in the cell counting kit 8. The expression of c-Myc and Cyclin D1 decreased dramatically in 3T3-L1 when Clock was silenced with short interfering RNA and was also decreased in fat tissue and adipose tissue-derived stem cells of Clock(Δ19) mice. Clock directly controls the expression of the components of Wnt signal transduction pathway, which was verified by serum shock, chromatin immunoprecipitation, Western blot, and quantitative real-time polymerase chain reaction (qRT-PCR). Furthermore, IWR-1, a Wnt signal pathway inhibitor, inhibited the cell cycle promotion by CLOCK, which was detected by cell viability assay, flow cytometry, and qRT-PCR. Therefore, CLOCK transcription control of Wnt signaling promotes cell cycle progression in 3T3-L1 preadipocytes. Clock inhibited the adipogenesis on day 2 in 3T3-L1 cells via Oil Red O staining and qRT-PCR detection and probably related to cellular differentiation. These data provide evidence that the circadian gene Clock regulates the proliferation of preadipocytes and affects adipogenesis. © 2016 IUBMB Life, 68(7):557-568, 2016. PMID:27194636

  3. Human c-fgr induces a monocyte-specific enzyme in NIH 3T3 cells

    SciTech Connect

    Inoue, Kazushi; Akiyama, Tetsu; Toyoshima, Kumao ); Wongsasant, Budsaba )

    1991-12-01

    The mutant c-fgr protein (p58{sup c-fgr/F523}) containing Phe-523 instead of Tyr-523 exhibited transforming activity in NIH 3T3 cells like other protein-tyrosine kinases of the src family, but normal p58{sup c-fgr} (p58{sup c-fgr/wt}) did not. The mutant protein showed tyrosine kinase activity threefold higher than that of the normal protein in vitro. Surprisingly, transfection of the normal c-fgr gene into NIH 3T3 cells resulted in induction of sodium fluoride (NaF)-sensitive {alpha}-naphthyl butyrate esterase ({alpha}-NBE), marker enzyme of cells of monocytic origin, which was not induced in v-src-, v-fgr-, or lyn-transfected NIH 3T3 cells. The NaF-sensitive {alpha}-NBE induced in c-fgr transfectants was shown by isoelectric focusing to have a pI of 5.2 to 5.4, a range which was the same as those for thioglycolate-induced murine peritoneal macrophages and 1{alpha}, 25-dihydroxyvitamin D{sub 3}-treated WEHI-3B cells. Immunoblotting studies with antophosphotyrosine antibodies revealed that 58-, 62-, 75-, 120-, 200-, and 230-kDa proteins were commonly phosphorylated at tyrosine residues in NIH 3T3 cells transfected with normal and mutated c-fgr, while 95-kDa protein was significantly phosphorylated at tyrosine residues in NIH 3T3 cells transfected with normal and mutated c-fgr, while 95-kDa protein was significantly phosphorylated at tyrosine residues in cells transfected with the mutated c-fgr. These findings suggest that tyrosine phosphorylation of specific cellular substrate proteins is important in induction of NaF-sensitive {alpha}-NBE and cell transformation by p58{sup c-fgr}.

  4. Induction of pyruvate carboxylase apoenzyme and holoenzyme in 3T3-L1 cells during differentiation

    PubMed Central

    Freytag, Svend O.; Utter, Merton F.

    1980-01-01

    The specific activity of pyruvate carboxylase [pyruvate:carbon-dioxide ligase (ADP-forming); EC 6.4.1.1] in 3T3-L1 cells increases approximately 20-fold when these cells differentiate to an adipocyte-like form [Mackall, J. C. & Lane, M. D. (1977) Biochem. Biophys. Res. Commun. 79, 720-725]. A specific antibody to the purified rat liver enzyme quantitatively precipitated pyruvate carboxylase from 3T3-L1 crude homogenates. Use of this immunological technique permitted us to demonstrate that the increase in pyruvate carboxylase activity is due to an increase in the intracellular concentration of the enzyme. The content of pyruvate carboxylase in differentiated 3T3-L1 cells is sufficiently high (1-2% of total protein) that the increase in this large protein (subunit Mr = 130,000) can be visualized when 3T3-L1 crude extracts are subjected to electrophoresis on sodium dodecyl sulfate/polyacrylamide gels. When 3T3-L1 cells differentiated in the presence of avidin, they contained less than 5% of the pyruvate carboxylase activity of cells that differentiated in the absence of avidin. However, the immunoprecipitable pyruvate carboxylase content of the avidin-treated cells was essentially the same as that of cells that differentiated without avidin. Full activity of the enzyme was rapidly restored in the avidin-treated cells upon the addition of excess biotin. The recovery of activity was closely correlated with the incorporation of [14C]biotin into immunoprecipitable pyruvate carboxylase. The rapidity with which the activity was restored and the insensitivity of the process to inhibitors of protein synthesis strongly suggest that the apoenzyme of pyruvate carboxylase accumulates during differentiation in the presence of avidin. Images PMID:6929488

  5. 3-T MRI safety assessments of magnetic dental attachments and castable magnetic alloys

    PubMed Central

    Miyata, K; Abe, Y; Ishii, T; Ishigami, T; Ohtani, K; Nagai, E; Ohyama, T; Umekawa, Y; Nakabayashi, S

    2015-01-01

    Objectives: To assess the safety of different magnetic dental attachments during 3-T MRI according to the American Society for Testing and Materials F2182-09 and F2052-06e1 standard testing methods and to develop a method to determine MRI compatibility by measuring magnetically induced torque. Methods: The temperature elevations, magnetically induced forces and torques of a ferromagnetic stainless steel keeper, a coping comprising a keeper and a cast magnetic alloy coping were measured on MRI systems. Results: The coping comprising a keeper demonstrated the maximum temperature increase (1.42 °C) for the whole-body-averaged specific absorption rate and was calculated as 2.1 W kg−1 with the saline phantom. All deflection angles exceeded 45°. The cast magnetic alloy coping had the greatest deflection force (0.33 N) during 3-T MRI and torque (1.015 mN m) during 0.3-T MRI. Conclusions: The tested devices showed minimal radiofrequency (RF)-induced heating in a 3-T MR environment, but the cast magnetic alloy coping showed a magnetically induced deflection force and torque approximately eight times that of the keepers. For safety, magnetic dental attachments should be inspected before and after MRI and large prostheses containing cast magnetic alloy should be removed. Although magnetic dental attachments may pose no great risk of RF-induced heating or magnetically induced torque during 3-T MRI, their magnetically induced deflection forces tended to exceed acceptable limits. Therefore, the inspection of such devices before and after MRI is important for patient safety. PMID:25785821

  6. Diagnosis of an imprinted-gene syndrome by a novel bioinformatics analysis of whole-genome sequences from a family trio.

    PubMed

    Bodian, Dale L; Solomon, Benjamin D; Khromykh, Alina; Thach, Dzung C; Iyer, Ramaswamy K; Link, Kathleen; Baker, Robin L; Baveja, Rajiv; Vockley, Joseph G; Niederhuber, John E

    2014-11-01

    Whole-genome sequencing and whole-exome sequencing are becoming more widely applied in clinical medicine to help diagnose rare genetic diseases. Identification of the underlying causative mutations by genome-wide sequencing is greatly facilitated by concurrent analysis of multiple family members, most often the mother-father-proband trio, using bioinformatics pipelines that filter genetic variants by mode of inheritance. However, current pipelines are limited to Mendelian inheritance patterns and do not specifically address disorders caused by mutations in imprinted genes, such as forms of Angelman syndrome and Beckwith-Wiedemann syndrome. Using publicly available tools, we implemented a genetic inheritance search mode to identify imprinted-gene mutations. Application of this search mode to whole-genome sequences from a family trio led to a diagnosis for a proband for whom extensive clinical testing and Mendelian inheritance-based sequence analysis were nondiagnostic. The condition in this patient, IMAGe syndrome, is likely caused by the heterozygous mutation c.832A>G (p.Lys278Glu) in the imprinted gene CDKN1C. The genotypes and disease status of six members of the family are consistent with maternal expression of the gene, and allele-biased expression was confirmed by RNA-Seq for the heterozygotes. This analysis demonstrates that an imprinted-gene search mode is a valuable addition to genome sequence analysis pipelines for identifying disease-causative variants. PMID:25614875

  7. Diagnosis of an imprinted-gene syndrome by a novel bioinformatics analysis of whole-genome sequences from a family trio

    PubMed Central

    Bodian, Dale L; Solomon, Benjamin D; Khromykh, Alina; Thach, Dzung C; Iyer, Ramaswamy K; Link, Kathleen; Baker, Robin L; Baveja, Rajiv; Vockley, Joseph G; Niederhuber, John E

    2014-01-01

    Whole-genome sequencing and whole-exome sequencing are becoming more widely applied in clinical medicine to help diagnose rare genetic diseases. Identification of the underlying causative mutations by genome-wide sequencing is greatly facilitated by concurrent analysis of multiple family members, most often the mother–father–proband trio, using bioinformatics pipelines that filter genetic variants by mode of inheritance. However, current pipelines are limited to Mendelian inheritance patterns and do not specifically address disorders caused by mutations in imprinted genes, such as forms of Angelman syndrome and Beckwith–Wiedemann syndrome. Using publicly available tools, we implemented a genetic inheritance search mode to identify imprinted-gene mutations. Application of this search mode to whole-genome sequences from a family trio led to a diagnosis for a proband for whom extensive clinical testing and Mendelian inheritance-based sequence analysis were nondiagnostic. The condition in this patient, IMAGe syndrome, is likely caused by the heterozygous mutation c.832A>G (p.Lys278Glu) in the imprinted gene CDKN1C. The genotypes and disease status of six members of the family are consistent with maternal expression of the gene, and allele-biased expression was confirmed by RNA-Seq for the heterozygotes. This analysis demonstrates that an imprinted-gene search mode is a valuable addition to genome sequence analysis pipelines for identifying disease-causative variants. PMID:25614875

  8. Fipronil promotes adipogenesis via AMPKα-mediated pathway in 3T3-L1 adipocytes.

    PubMed

    Sun, Quancai; Qi, Weipeng; Yang, Jeremy J; Yoon, Kyong Sup; Clark, John M; Park, Yeonhwa

    2016-06-01

    Emerging evidence suggests that organochlorine, organophosphorus and neonicotinoid insecticide exposure may be linked to the development of obesity and type 2 diabetes. However, there is no knowledge of the potential influence of fipronil, which belongs to the phenylpyrazole chemical family, on obesity. Thus, the goal of this study was to determine the role of fipronil in adipogenesis using 3T3-L1 adipocytes. Fipronil treatment, at 10 μM, increased fat accumulation in 3T3-L1 adipocytes as well as promoted key regulators of adipocyte differentiation (CCAAT/enhancer-binding protein α and peroxisome proliferator-activated receptor gamma-γ), and key regulators of lipogenesis (acetyl-CoA carboxylase and fatty acid synthase). The activation of AMPKα with 5-aminoimidazole-4-carboxamide ribonucleotide (AICAR) abolished effects of fipronil on increased adipogenesis. These results suggest that fipronil alters adipogenesis and results in increased lipid accumulation through a AMPKα-mediated pathway. PMID:27103584

  9. Genome sequence of the organohalide-respiring Dehalogenimonas alkenigignens type strain (IP3-3(T)).

    PubMed

    Key, Trent A; Richmond, Dray P; Bowman, Kimberly S; Cho, Yong-Joon; Chun, Jongsik; da Costa, Milton S; Rainey, Fred A; Moe, William M

    2016-01-01

    Dehalogenimonas alkenigignens IP3-3(T) is a strictly anaerobic, mesophilic, Gram negative staining bacterium that grows by organohalide respiration, coupling the oxidation of H2 to the reductive dehalogenation of polychlorinated alkanes. Growth has not been observed with any non-polyhalogenated alkane electron acceptors. Here we describe the features of strain IP3-3(T) together with genome sequence information and its annotation. The 1,849,792 bp high-quality-draft genome contains 1936 predicted protein coding genes, 47 tRNA genes, a single large subunit rRNA (23S-5S) locus, and a single, orphan, small unit rRNA (16S) locus. The genome contains 29 predicted reductive dehalogenase genes, a large majority of which lack cognate genes encoding membrane anchoring proteins. PMID:27340512

  10. MANTIS-3T: a low-cost light-weight turreted spectral sensor

    NASA Astrophysics Data System (ADS)

    Dirbas, Joseph; Mireles, Tony; Davies, Adam; Schoonmaker, Jon; Lovett, Alexander R.

    2005-05-01

    PAR Government Systems Corporation (PAR) has developed a low-cost, low-weight, low-profile, mission-adaptable multispectral imaging system utilizing mass-produced commercial off-the-shelf (COTS) components, for the purpose of providing continuous real-time multispectral data collection for mine counter measures (MCM), intelligence, surveillance, and reconnaissance study applications aboard low-cost, light manned and unmanned aircraft platforms. The mission adaptable narrowband tunable imaging system (MANTIS) has been integrated into a small 5" turret currently employed on a variety of small UAV platforms. The turreted MANTIS (MANTIS-3T) provides remote operator control to adjust gain, exposure, and pointing commands. The MANTIS-3T sensor will be used to collect imagery over calibration and test targets. Integration strategies and planned data collections are presented.

  11. The Holy Grail in diagnostic neuroradiology: 3T or 3D?

    PubMed Central

    Pouwels, Petra J. W.; Wattjes, Mike P.

    2010-01-01

    Many technical developments keep occurring in the field of MRI that could benefit image acquisition in the field of diagnostic neuroradiology. While there is much focus on the potential advantages of 3T and higher field strengths, it is often unclear whether these are cosmetic only, or convey clinically relevant diagnostic value. The increased signal-to-noise at 3T is certainly beneficial in different ways particularly for the acquisition of isotropic 3D sequences like FLAIR. Single-slab 3D sequences can now be obtained with multiple contrasts in clinically attainable data acquisition times and could revolutionize MRI to evolve into a fundamentally multi-planar technique, rather similar to what has happened with the introduction of multi-detector row CT. PMID:21181406

  12. Expression of Nanog gene promotes NIH3T3 cell proliferation

    SciTech Connect

    Zhang Jingyu; Wang Xia; Chen Bing; Suo Guangli; Zhao Yanhong; Duan Ziyuan; Dai Jianwu . E-mail: jwdai@genetics.ac.cn

    2005-12-16

    Cells are the functional elements in tissue engineering and regenerative medicine. A large number of cells are usually needed for these purposes. However, there are numbers of limitations for in vitro cell proliferation. Nanog is an important self-renewal determinant in embryonic stem cells. However, it remains unknown whether Nanog will influence the cell cycle and cell proliferation of mature cells. In this study, we expressed Nanog in NIH3T3 cells and showed that expression of Nanog in NIH3T3 promoted cells to enter into S phase and enhanced cell proliferation. This suggests that Nanog gene might function in a similar fashion in mature cells as in ES cells. In addition, it may provide an approach for in vitro cell expansion.

  13. Recommended protocol for the BALB/c 3T3 cell transformation assay.

    PubMed

    Sasaki, Kiyoshi; Bohnenberger, Susanne; Hayashi, Kumiko; Kunkelmann, Thorsten; Muramatsu, Dai; Phrakonkham, Pascal; Poth, Albrecht; Sakai, Ayako; Salovaara, Susan; Tanaka, Noriho; Thomas, B Claire; Umeda, Makoto

    2012-04-11

    The present protocol has been developed for the BALB/c 3T3 cell transformation assay (CTA), following the prevalidation study coordinated by the European Centre for the Validation of Alternative Methods (ECVAM) and reported in this issue (Tanaka et al. [16]). Based upon the experience gained from this effort and as suggested by the Validation Management Team (VMT), some acceptance and assessment criteria have been refined compared to those used during the prevalidation study. The present protocol thus describes cell culture maintenance, the dose-range finding (DRF) experiment and the transformation assay, including cytotoxicity and morphological transformation evaluation. Use of this protocol and of the associated photo catalogue included in this issue (Sasaki et al. [17]) is recommended for the future conduct of the BALB/c 3T3 CTA. PMID:22212201

  14. A novel RF resonator for human-body MRI at 3 T

    NASA Astrophysics Data System (ADS)

    Son, Hyeok-Woo; Cho, Young-Ki; Yoo, Hyoungsuk

    2014-03-01

    A square-slot-loaded (SSL) radio-frequency (RF) resonator using a microstrip transmission line (MTL) is designed for human-body magnetic resonance imaging (MRI) at 3 T MRI. The SSL RF resonator shows improved RF magnetic fields resulting in more homogenous fields near the center of the phantom than traditional RF resonators using MTL. A multichannel body coil using the SSL RF resonators is also simulated and provides improved parallel excitation performance. In addition, RF shimming for homogenization can be effectively controlled by adjusting the inputs to the eight resonators. Numerical results are obtained by using a spherical phantom and a realistic human-body model at 3 T to calculate the B {1/+} fields.

  15. Human papillomavirus type 16 DNA-induced malignant transformation of NIH 3T3 cells

    SciTech Connect

    Yasumoto, S.; Burkhardt, A.L.; Doniger, J.; DiPaolo, J.A.

    1986-02-01

    A biological function for human papillomavirus 16 (HPV 16) DNA was demonstrated by transformation of NIH 3T3 cells. HPV 16 DNA has been found frequently in genital cancer and has been classified as a papillomavirus on the basis of DNA homology. A recombinant HPV 16 DNA (pSHPV16d), which contains a head-to-tail dimer of the full-length HPV 16 genome, induced morphologic transformation; the transformed cells were tumorigenic in nude mice. Expression of transforming activity was unique because of the long latency period (more than 4 weeks) required for induction of morphologic transformation and because the transfected DNA existed primarily in a multimeric form with some rearrangement. Furthermore, virus-specific RNAs were expressed in the transformants. The transformation of NIH 3T3 cells provides a model for analyzing the functions of HPV 16, which is associated with cervical carcinomas.

  16. Polyclonal B-cell stimulation by L3T4+ T cells in experimental leishmaniasis.

    PubMed Central

    Lohoff, M; Matzner, C; Röllinghoff, M

    1988-01-01

    The well-established polyclonal B-cell stimulation in the lymphoid organs in mice infected with Leishmania major is thought to be dependent on T cells. Here we present clear experimental evidence that this is indeed the case by showing that BALB/c-derived, L3T4-positive L. major-specific T cells induce syngeneic B cells to polyclonal proliferation and immunoglobulin production. PMID:3260894

  17. Philips 3T Intera Magnetic Resonance Imaging System and Upgrade of existing MRI equipment

    SciTech Connect

    Evanochko, William T

    2004-05-14

    The objective of this proposal was twofold. First, upgrade existing MRI equipment, specifically a research 4.1T whole-body system. Second, purchase a clinical, state-of-the-art 3T MRI system tailored specifically to cardiovascular and neurological applications. This project was within the guidelines of ''Medical Applications and Measurement Science''. The goals were: [1] to develop beneficial applications of magnetic resonance imaging; [2] discover new applications of MR strategies for medical research; and [2] apply them for clinical diagnosis. Much of this proposal searched for breakthroughs in this noninvasive and nondestructive imaging technology. Finally, this proposal's activities focused on research in the basic science of chemistry, biochemistry, physics, and engineering as applied to bioengineering. The centerpiece of this grant was our 4.1T ultra-high field whole-body nuclear magnetic resonance system and the newly acquired state-of-the-art, heart and head dedicated 3T clinical MRI system. We have successfully upgraded the equipment for the 4.1T system so that it is now state-of-the-art with new gradient and radio frequency amplifiers. We also purchase a unique In Vivo EKG monitoring unit that will permit tracking clinical quality EKG signals while the patient is in a high field MR scanner. Important upgrades of a peripheral vascular coil and a state-of-the-art clinical workstation for processing complex heart images were implemented. The most recent acquisition was the purchase of a state-of-the-art Philips 3T Intera clinical MRI system. This system is unique in that the magnet is only 5 1/2 feet long compare to over 12 feet long magnet of our 4.1T MRI system. The 3T MRI system is fully functional and its use and applications are already greatly benefiting the UAB with 200-300 micron resolution brain images and diagnostic quality MR angiography of coronary arteries in less than 5 minutes.

  18. Ultrasound associated uptake of chitosan nanoparticles in MC3T3-E1 cells

    NASA Astrophysics Data System (ADS)

    Wu, Junyi

    Chitosan is a natural linear polysaccharide that has been well known for its applications in drug delivery system due to its unique physicochemical and biological properties. However, challenges still remain for it to become a fully realized therapeutic agent. In this study, we investigated the uptake of chitosan nanoparticles (CNP) under the ultrasound stimulation, using a model cell culture system (MC3T3-E1 mouse pre-osteoblasts). The CNP were fabricated by an ionic gelation method and were lyophilized prior to characterization and delivery to cells. Particle size and zeta potential were measured using Dynamic Light Scattering (DLS); the efficiency of chitosan complexation was measured using the ninhydrin assay. Cytotoxicity was examined by neutral red assay within 48 hours after delivery. The effect of ultrasound (US) on the efficiency of nanoparticle delivery to the MC3T3-E1 cells was examined at 1MHz and at either 1 or 2 W/cm2. Fluorescein isothiocyanate (FITC)-conjugated-CNP were used to visualize the internalized particles within the cytosol. The uptake of FITC-CNP exhibits a dose and time dependent effect, a strong FITC fluorescence was detected at the concentration of 500microg/mL under fluorescence microscope. Ultrasound assisted uptake of FITC-CNP performed a significant positive effect at 2W/cm2 with 60s of ultrasound exposure time. CNP displayed a slightly decrease in cell viability from 25microg/mL to 100microg/mL, while higher concentration of CNP facilitates the proliferation of MC3T3-E1 cells. Less than 10% of reduction in cell viability was observed for US at 1W/cm2 and 2W/cm2 with 30s and 60s of exposure time, which suggest a mild effect of US to MC3T3-E1 cell line.

  19. Extract of Chaga mushroom (Inonotus obliquus) stimulates 3T3-L1 adipocyte differentiation.

    PubMed

    Joo, Jeong In; Kim, Dong Hyun; Yun, Jong Won

    2010-11-01

    Chaga mushroom (Inonotus obliquus) has long been used as a folk medicine due to its numerous biological functions such as antibacterial, antiallergic, antiinflammatory and antioxidative activities. In the present study, it was found that the I. obliquus hot water extract (IOWE) activated adipogenesis of 3T3-L1 preadipocytes. Even in the absence of adipogenic stimuli by insulin, the IOWE strongly induced adipogenesis of 3T3-L1 preadipocytes. The major constituent of IOWE was glucose-rich polysaccharides with a molecular mass of 149  kDa. IOWE enhanced the differentiation of 3T3-L1 preadipocytes, increasing TG (triacylglycerol) accumulation that is critical for acquisition of the adipocyte phenotype, in a dose-dependent manner. IOWE stimulated gene expression of C/EBPα (CCAAT/enhancer-binding protein α) and PPARγ (peroxisome proliferator-activated receptors γ) during adipocyte differentiation, and induced the expression of PPARγ target genes such as aP2 (adipocyte protein 2), LPL (lipoprotein lipase) and CD36 (fatty acid translocase). Immunoblot analysis revealed that IOWE increased the expression of adipogenic makers such as PPARγ and GLUT4 (glucose transporter 4). The luciferase reporter assay demonstrated that IOWE did not exhibit PPARγ ligand activity. Although these results require further investigation, the ability of natural mushroom product to increase PPARγ transcriptional activities may be expected to be therapeutic targets for dyslipidemia and type 2 diabetes. PMID:21031614

  20. Semicarbazide-sensitive amine oxidase activation promotes adipose conversion of 3T3-L1 cells.

    PubMed Central

    Mercier, N; Moldes, M; El Hadri, K; Fève, B

    2001-01-01

    Semicarbazide-sensitive amine oxidase (SSAO) is an amine oxidase related to the copper-containing amine oxidase family. The tissular form of SSAO is located at the plasma membrane, and is mainly expressed in vascular smooth muscle cells and adipocytes. Recent studies have suggested that SSAO could activate glucose transport in fat cells. In the present work, we investigated the potential role of a chronic SSAO activation on adipocyte maturation of the 3T3-L1 pre-adipose cell line. Exposure of post-confluent 3T3-L1 pre-adipocytes to methylamine, a physiological substrate of SSAO, promoted adipocyte differentiation in a time- and dose-dependent manner. This effect could be related to SSAO activation, since it was antagonized in the presence of the SSAO inhibitor semicarbazide, but not in the presence of the monoamine oxidase inhibitor pargyline. In addition, methylamine-induced adipocyte maturation was mimicked by 3T3-L1 cell treatment with other SSAO substrates. Finally, the large reversion of methylamine action by catalase indicated that hydrogen peroxide generated by SSAO was involved, at least in part, in the modulation of adipocyte maturation. Taken together, our results suggest that SSAO may contribute to the control of adipose tissue development. PMID:11513731

  1. Fat-suppression techniques for 3-T MR imaging of the musculoskeletal system.

    PubMed

    Del Grande, Filippo; Santini, Francesco; Herzka, Daniel A; Aro, Michael R; Dean, Cooper W; Gold, Garry E; Carrino, John A

    2014-01-01

    Fat suppression is an important technique in musculoskeletal imaging to improve the visibility of bone-marrow lesions; evaluate fat in soft-tissue masses; optimize the contrast-to-noise ratio in magnetic resonance (MR) arthrography; better define lesions after administration of contrast material; and avoid chemical shift artifacts, primarily at 3-T MR imaging. High-field-strength (eg, 3-T) MR imaging has specific technical characteristics compared with lower-field-strength MR imaging that influence the use and outcome of various fat-suppression techniques. The most commonly used fat-suppression techniques for musculoskeletal 3-T MR imaging include chemical shift (spectral) selective (CHESS) fat saturation, inversion recovery pulse sequences (eg, short inversion time inversion recovery [STIR]), hybrid pulse sequences with spectral and inversion-recovery (eg, spectral adiabatic inversion recovery and spectral attenuated inversion recovery [SPAIR]), spatial-spectral pulse sequences (ie, water excitation), and the Dixon techniques. Understanding the different fat-suppression options allows radiologists to adopt the most appropriate technique for their clinical practice. PMID:24428292

  2. Ramie Leaf Extracts Suppresses Adipogenic Differentiation in 3T3-L1 Cells and Pig Preadipocytes

    PubMed Central

    Lee, Joomin; Kim, Ah-Ra; Lee, Jae-Joon

    2016-01-01

    The present study was carried out to evaluate the anti-obesity effect of different concentrations of extracts of hot air-dried ramie leaf (HR) and freeze-dried ramie leaf (FR) in 3T3-L1 cells and pig preadipocytes. To analyze the effect on cell proliferation, cells were treated with 25 μg/mL or 100 μg/mL HR or FR extract for 2 days. Cell differentiation was evaluated by measuring glycerol-3-phosphate dehydrogenase and lipoprotein lipase (LPL) activities and intracellular triglyceride content. Treatment with either HR or FR extracts inhibited the proliferation of 3T3-L1 cells and pig preadipocytes in a dose-dependent manner. HR extract treatment inhibited the differentiation of both cell types more effectively than FR treatment. The extent of triglyceride accumulation decreased significantly in both cells following either HR or FR treatment. Furthermore, LPL activity significantly decreased after treatment with HR or FR extract. These results indicated that HR and FR extracts may inhibit proliferation and differentiation of 3T3-L1 cells and pig preadipocytes. Further studies are needed to explore the anti-obesity effect of HR and FR extracts. PMID:26954122

  3. WEHI-3 cells inhibit adipocyte differentiation in 3T3-L1 cells

    SciTech Connect

    Lai, Jing; Liu, Gexiu; Yan, Guoyao; He, Dongmei; Zhou, Ying; Chen, Shengting

    2015-06-26

    By investigating the anti-adipogenic effects of WEHI-3 cells – a murine acute myelomonocytic leukemia cell line – we sought to improve the efficiency of hematopoietic stem cell transplantation (HSCT). Analysis of Oil Red O staining and the expression of adipogenic genes, including PPARγ, C/EBPα, FAS and LPL, indicated that WEHI-3 cells significantly inhibited 3T3-L1 mouse preadipocyte cells from differentiating into adipocytes. In vivo, fat vacuoles in mice injected with WEHI-3 cells were also remarkably reduced in the murine bone marrow pimelosis model. Moreover, the key gene in the Rho signaling pathway, ROCKII, and the key gene in the Wnt signaling pathway, β-catenin, were both upregulated compared with the control group. siRNA-mediated knockdown of ROCKII and β-catenin reversed these WEHI-3-mediated anti-adipogenic effects. Taken together, these data suggest that WEHI-3 cells exert anti-adipogenic effects and that both ROCKII and β-catenin are involved in this process. - Highlights: • WEHI-3, an acute myelomonocytic leukemia cell line, inhibited 3T3-L1 preadipocyte from differentiating into adipocyte. • WEHI-3 cells can arrest 3T3-L1 cells in G0/G1 phase by secreting soluble factors and thus inhibit their proliferation. • WEHI-3 cells reduced bone marrow pimelosis in the murine model. • Both ROCKII and β-catenin were involved in the WEHI-3-mediated anti-adipogenic effects.

  4. Effect of Mangiferin and Mahanimbine on Glucose Utilization in 3T3-L1 cells

    PubMed Central

    Kumar, B Dinesh; Krishnakumar, K; Jaganathan, Saravana Kumar; Mandal, Mahitosh

    2013-01-01

    Background: Stem barks of Mangifera indica contain a rich content of mangiferin (xanthone glucoside), whereas Murraya koenigii leaves contain rich sources of mahanimbine (carbazole alkaloid) and used traditionally for the treatment of diabetes. Objective: To investigate the effects of mangiferin (xanthone glucoside) and mahanimbine (carbazole alkaloid) on glucose utilization in 3T3-L1 cells. Materials and Methods: Mangiferin was isolated from stem barks of Mangifera indica and mahanimbine was isolated from Murraya koenigii leaves. These isolated compounds were subjected to MTT assay and glucose utilization test with 3T3-L1 cells. Results: Treatment of the 3T3-L1 cells with mangiferin and mahanimbine increased the glucose utilization in a dose-dependent manner. At a concentration of 1 mM, mangniferin showed 2-fold increase in glucose utilization compared with untreated control. In case of mahanimbine, the observed effect at 1 mM was almost equivalent to positive control (insulin at 1 μM). Moreover, MTT assay showed that both of these compounds were less toxic at a concentration of 1 mM (nearly 75% cells are viable). Conclusion: The present results indicated that these natural products (mangiferin and mahanimbine) exhibited potential ethnomedical uses in management of diabetes. PMID:23661997

  5. Fat-Suppression Techniques for 3-T MR Imaging of the Musculoskeletal System1

    PubMed Central

    Del Grande, Filippo; Santini, Francesco; Herzka, Daniel A.; Aro, Michael R.; Dean, Cooper W.; Gold, Garry E.; Carrino, John A.

    2015-01-01

    Fat suppression is an important technique in musculoskeletal imaging to improve the visibility of bone-marrow lesions; evaluate fat in soft-tissue masses; optimize the contrast-to-noise ratio in magnetic resonance (MR) arthrography; better define lesions after administration of contrast material; and avoid chemical shift artifacts, primarily at 3-T MR imaging. High-field-strength (eg, 3-T) MR imaging has specific technical characteristics compared with lower-field-strength MR imaging that influence the use and outcome of various fat-suppression techniques. The most commonly used fat-suppression techniques for musculoskeletal 3-T MR imaging include chemical shift (spectral) selective (CHESS) fat saturation, inversion recovery pulse sequences (eg, short inversion time inversion recovery [STIR]), hybrid pulse sequences with spectral and inversion-recovery (eg, spectral adiabatic inversion recovery and spectral attenuated inversion recovery [SPAIR]), spatial-spectral pulse sequences (ie, water excitation), and the Dixon techniques. Understanding the different fat-suppression options allows radiologists to adopt the most appropriate technique for their clinical practice. PMID:24428292

  6. Rapamycin inhibits clonal expansion and adipogenic differentiation of 3T3-L1 cells.

    PubMed Central

    Yeh, W C; Bierer, B E; McKnight, S L

    1995-01-01

    Differentiating 3T3-L1 cells express an immunophilin early during the adipocyte conversion program as described in this issue [Yeh, W.-C., Li, T.-K., Bierer, B. E. & McKnight, S. L. (1995) Proc. Natl. Acad. Sci. USA 92, 11081-11085]. The temporal expression profile of this protein, designated FK506-binding protein (FKBP) 51, is concordant with the clonal-expansion period undertaken by 3T3-L1 cells after exposure to adipogenic hormones. Having observed FKBP51 synthesis early during adipogenesis, we tested the effects of three immunosuppressive drugs--cyclosporin A, FK506, and rapamycin--on the terminal-differentiation process. Adipocyte conversion was not affected by either cyclosporin A or FK506 and yet was significantly reduced by rapamycin at drug concentrations as low as 10 nM. Clonal expansion was impeded in drug-treated cultures, as was the accumulation of cytoplasmic lipid droplets normally seen late during differentiation. Rapamycin treatment likewise inhibited the expression of CCAAT/enhancer binding protein alpha, a transcription factor required for 3T3-L1 cell differentiation. All three of these effects were reversed by high FK506 concentrations, indicating that the operative inhibitory event was mediated by an immunophilin-rapamycin complex. Images Fig. 1 Fig. 2 Fig. 3 PMID:7479942

  7. Endoplasmic reticulum stress suppresses lipin-1 expression in 3T3-L1 adipocytes

    SciTech Connect

    Takahashi, Nobuhiko; Hiranaka, Natsumi; Suzuki, Takeshi; Yui, Tomoo; Akanuma, Masayoshi; Kanazawa, Kaoru; Yoshida, Mika; Naito, Sumiyoshi; Fujiya, Mikihiro; Kohgo, Yutaka

    2013-02-01

    Highlights: ► Lipin-1 involves lipid metabolism, adipocyte differentiation, and inflammation. ► Adipose lipin-1 expression is reduced in obesity. ► ER stress suppresses lipin-1 expression in 3T3-L1 adipocytes. ► Activation of PPAR-γ recovers ER stress-induced lipin-1 reduction. -- Abstract: Lipin-1 plays crucial roles in the regulation of lipid metabolism and cell differentiation in adipocytes. In obesity, adipose lipin-1 mRNA expression is decreased and positively correlated with systemic insulin sensitivity. Amelioration of the lipin-1 depletion might be improved dysmetabolism. Although some cytokines such as TNF-α and interleukin-1β reduces adipose lipin-1 expression, the mechanism of decreased adipose lipin-1 expression in obesity remains unclear. Recently, endoplasmic reticulum (ER) stress is implicated in the pathogenesis of obesity. Here we investigated the role of ER stress on the lipin-1 expression in 3T3-L1 adipocytes. We demonstrated that lipin-1 expression was suppressed by the treatment with ER stress inducers (tunicamycin and thapsigargin) at transcriptional level. We also showed that constitutive lipin-1 expression could be maintained by peroxisome proliferator-activated receptor-γ in 3T3-L1 adipocytes. Activation of peroxisome proliferator-activated receptor-γ recovered the ER stress-induced lipin-1 suppression. These results suggested that ER stress might be involved in the pathogenesis of obesity through lipin-1 depletion.

  8. Sclerostin Enhances Adipocyte Differentiation in 3T3-L1 Cells.

    PubMed

    Ukita, Mayumi; Yamaguchi, Taihiko; Ohata, Noboru; Tamura, Masato

    2016-06-01

    Sclerostin, a secreted protein encoded by the Sost gene, is produced by osteocytes and is inhibited by osteoblast differentiation and bone formation. Recently, a functional association between bone and fat tissue has been suggested, and a correlation between circulating sclerostin levels and lipid metabolism has been reported in humans. However, the effects of sclerostin on adipogenesis remain unexplored. In the present study, we examined the role of sclerostin in regulating adipocyte differentiation using 3T3-L1 preadipocytes. In these cells, sclerostin enhanced adipocyte-specific gene expression and the accumulation of lipid deposits. Sclerostin also upregulated CCAAT/enhancer binding protein β expression but not cell proliferation and caspase-3/7 activities. Sclerostin also attenuated canonical Wnt3a-inhibited adipocyte differentiation. Recently, the transcriptional modulator TAZ has been involved in the canonical Wnt signaling pathway. Sclerostin reduced TAZ-responsive transcriptional activity and TAZ-responsive gene expression. Transfection of 3T3-L1 cells with TAZ siRNA increased the lipid deposits and adipogenic gene expression. These results show that sclerostin upregulates adipocyte differentiation in 3T3-L1 cells, suggesting a possible role for the osteocyte-derived sclerostin as a regulator of fat metabolism and as a reciprocal regulator of bone and adipose tissues metabolism. J. Cell. Biochem. 117: 1419-1428, 2016. © 2015 Wiley Periodicals, Inc. PMID:26553151

  9. Ginkgolide C Suppresses Adipogenesis in 3T3-L1 Adipocytes via the AMPK Signaling Pathway

    PubMed Central

    Liou, Chian-Jiun; Lai, Xuan-Yu; Chen, Ya-Ling; Wang, Chia-Ling; Wei, Ciao-Han; Huang, Wen-Chung

    2015-01-01

    Ginkgolide C, isolated from Ginkgo biloba leaves, is a flavone reported to have multiple biological functions, from decreased platelet aggregation to ameliorating Alzheimer disease. The study aim was to evaluate the antiadipogenic effect of ginkgolide C in 3T3-L1 adipocytes. Ginkgolide C was used to treat differentiated 3T3-L1 cells. Cell supernatant was collected to assay glycerol release, and cells were lysed to measure protein and gene expression related to adipogenesis and lipolysis by western blot and real-time PCR, respectively. Ginkgolide C significantly suppressed lipid accumulation in differentiated adipocytes. It also decreased adipogenesis-related transcription factor expression, including peroxisome proliferator-activated receptor and CCAAT/enhancer-binding protein. Furthermore, ginkgolide C enhanced adipose triglyceride lipase and hormone-sensitive lipase production for lipolysis and increased phosphorylation of AMP-activated protein kinase (AMPK), resulting in decreased activity of acetyl-CoA carboxylase for fatty acid synthesis. In coculture with an AMPK inhibitor (compound C), ginkgolide C also improved activation of sirtuin 1 and phosphorylation of AMPK in differentiated 3T3-L1 cells. The results suggest that ginkgolide C is an effective flavone for increasing lipolysis and inhibiting adipogenesis in adipocytes through the activated AMPK pathway. PMID:26413119

  10. Traditional Herbal Formula Oyaksungi-San Inhibits Adipogenesis in 3T3-L1 Adipocytes

    PubMed Central

    Seo, Chang-Seob; Shin, Hyeun-Kyoo

    2015-01-01

    Background. Oyaksungi-san (OYSGS) is a herbal formula that has been used for treating cardiovascular diseases in traditional Asian medicine. Here, we investigated the antiadipogenic effect of OYSGS extract in 3T3-L1 adipose cells. Methods. 3T3-L1 preadipocytes were differentiated into adipocytes with or without OYSGS. After differentiation, we measured Oil Red O staining, glycerol-3-phosphate dehydrogenase (GPDH) activity, leptin production, mRNA, and protein levels of adipogenesis-related factors. Results. OYSGS extract dramatically inhibited intracellular lipid accumulation in the differentiated adipocytes. It also significantly suppressed the (GPDH) activity, triglyceride (TG) content, and leptin production by reducing the expression of adipogenesis-related genes including lipoprotein lipase, fatty acid binding protein 4, CCAAT/enhancer-binding protein-alpha (C/EBP-α), and peroxisome proliferator-activated receptor gamma (PPAR-γ). Furthermore, OYSGS clearly enhanced phosphorylation of AMP-activated protein kinase (AMPK) as well as its substrate acetyl CoA (ACC) carboxylase. Conclusions. Our results demonstrate that OYSGS negatively controls TG accumulation in 3T3-L1 adipocytes. We suggest antiadipogenic activity of OYSGS and its potential benefit in preventing obesity. PMID:25802547

  11. Cardiovascular magnetic resonance imaging of isolated perfused pig hearts in a 3T clinical MR scanner

    PubMed Central

    Chiribiri, Amedeo; Ishida, Masaki; Morton, Geraint; Paul, Matthias; Hussain, Shazia T.; Bigalke, Boris; Perera, Divaka; Schaeffter, Tobias; Nagel, Eike

    2012-01-01

    Purpose An isolated perfused pig heart model has recently been proposed for the development of novel methods in standard clinical magnetic resonance (MR) scanners. The original set-up required the electrical system to be within the safe part of the MR-room, which introduced significant background noise. The purpose of the current work was to refine the system to overcome this limitation so that all electrical parts are completely outside the scanner room. Methods Four pig hearts were explanted under terminal anaesthesia from large white cross landrace pigs. All hearts underwent cardiovascular magnetic resonance (CMR) scanning in the MR part of a novel combined 3T MR and x-ray fluoroscopy (XMR) suite. CMR scanning included real-time k-t SENSE functional imaging, k-t SENSE accelerated perfusion imaging and late gadolinium enhancement imaging. Interference with image quality was assessed by spurious echo imaging and compared to noise levels acquired while operating the electrical parts within the scanner room. Results Imaging was performed successfully in all hearts. The system proved suitable for isolated heart perfusion in a novel 3T XMR suite. No significant additional noise was introduced into the scanner room by our set-up. Conclusions We have substantially improved a previous version of an isolated perfused pig heart model and made it applicable for MR imaging in a state of the art clinical 3T XMR imaging suite. The use of this system should aid novel CMR sequence development and translation into clinical practice. PMID:24265875

  12. Coronary artery anomalies and variants: technical feasibility of assessment with coronary MR angiography at 3 T.

    PubMed

    Gharib, Ahmed M; Ho, Vincent B; Rosing, Douglas R; Herzka, Daniel A; Stuber, Matthias; Arai, Andrew E; Pettigrew, Roderic I

    2008-04-01

    The purpose of this study was to prospectively use a whole-heart three-dimensional (3D) coronary magnetic resonance (MR) angiography technique specifically adapted for use at 3 T and a parallel imaging technique (sensitivity encoding) to evaluate coronary arterial anomalies and variants (CAAV). This HIPAA-compliant study was approved by the local institutional review board, and informed consent was obtained from all participants. Twenty-two participants (11 men, 11 women; age range, 18-62 years) were included. Ten participants were healthy volunteers, whereas 12 participants were patients suspected of having CAAV. Coronary MR angiography was performed with a 3-T MR imager. A 3D free-breathing navigator-gated and vector electrocardiographically-gated segmented k-space gradient-echo sequence with adiabatic T2 preparation pulse and parallel imaging (sensitivity encoding) was used. Whole-heart acquisitions (repetition time msec/echo time msec, 4/1.35; 20 degrees flip angle; 1 x 1 x 2-mm acquired voxel size) lasted 10-12 minutes. Mean examination time was 41 minutes +/- 14 (standard deviation). Findings included aneurysms, ectasia, arteriovenous fistulas, and anomalous origins. The 3D whole-heart acquisitions developed for use with 3 T are feasible for use in the assessment of CAAV. PMID:18372470

  13. Ramie Leaf Extracts Suppresses Adipogenic Differentiation in 3T3-L1 Cells and Pig Preadipocytes.

    PubMed

    Lee, Joomin; Kim, Ah-Ra; Lee, Jae-Joon

    2016-09-01

    The present study was carried out to evaluate the anti-obesity effect of different concentrations of extracts of hot air-dried ramie leaf (HR) and freeze-dried ramie leaf (FR) in 3T3-L1 cells and pig preadipocytes. To analyze the effect on cell proliferation, cells were treated with 25 μg/mL or 100 μg/mL HR or FR extract for 2 days. Cell differentiation was evaluated by measuring glycerol-3-phosphate dehydrogenase and lipoprotein lipase (LPL) activities and intracellular triglyceride content. Treatment with either HR or FR extracts inhibited the proliferation of 3T3-L1 cells and pig preadipocytes in a dose-dependent manner. HR extract treatment inhibited the differentiation of both cell types more effectively than FR treatment. The extent of triglyceride accumulation decreased significantly in both cells following either HR or FR treatment. Furthermore, LPL activity significantly decreased after treatment with HR or FR extract. These results indicated that HR and FR extracts may inhibit proliferation and differentiation of 3T3-L1 cells and pig preadipocytes. Further studies are needed to explore the anti-obesity effect of HR and FR extracts. PMID:26954122

  14. 17 CFR 240.15b7-3T - Operational capability in a Year 2000 environment.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 17 Commodity and Securities Exchanges 3 2011-04-01 2011-04-01 false Operational capability in a Year 2000 environment. 240.15b7-3T Section 240.15b7-3T Commodity and Securities Exchanges SECURITIES... § 240.15b7-3T Operational capability in a Year 2000 environment. (a) This section applies to...

  15. EPAS1 promotes adipose differentiation in 3T3-L1 cells.

    PubMed

    Shimba, Shigeki; Wada, Taira; Hara, Shuntaro; Tezuka, Masakatsu

    2004-09-24

    Adipose differentiation is regulated by several transcription factors, such as the CAAT/enhancer-binding protein family and peroxisome proliferator activator (PPAR) gamma2. Several recent studies have shown that the basic helix-loop-helix-PAS superfamily is also involved in the regulation of adipose differentiation. In this study, we investigated the roles played by EPAS1 (endothelial PAS domain protein 1) in adipogenesis. EPAS1, also referred to as hypoxia-inducible factor 2alpha, is a transcription factor known to play essential roles in catecholamine homeostasis, vascular remodeling, and the maintenance of reactive oxygen species, and so forth. During adipose differentiation in 3T3-L1 cells, the level of EPAS1 mRNA began to increase 6 days after the induction, and EPAS1 was highly expressed in differentiated cells. To examine whether EPAS1 is involved in adipogenesis, we first isolated stable clones from 3T3-L1 cells in which we could induce the expression of an EPAS1 C-terminal deletion mutant (designated EPAS1-(1-485)) with the insect hormone. The induction of EPAS1-(1-485) allowed the cells to accumulate only minimum amounts of intracellular lipid droplets. Consistent with the morphological observations, a minimum amount of aP2 and PPARgamma2 mRNA was induced in the EPAS1-(1-485) cells. We then examined whether or not EPAS1 was able to promote adipogenesis in NIH 3T3 cells, a relatively nonadipogenic cell line. Overexpression of EPAS1 in NIH 3T3 cells induced a significant amount of lipid accumulation compared with that of the control cells in the presence of the PPARgamma ligand. The results were also confirmed by measuring the expression of adipocyte-related genes. Adenovirus-mediated EPAS1-(1-485) expression resulted in the reduction of basal and insulin-dependent glucose transport in 3T3-L1 adipocytes. The mechanism involved the transcriptional regulation of GLUT1, GLUT4, and IRS3 expression by EPAS1. Taken together, these results suggest that EPAS1 plays

  16. Development of a Polysilicon Process Based on Chemical Vapor Deposition of Dichlorosilane in an Advanced Siemen's Reactor

    NASA Technical Reports Server (NTRS)

    Arevidson, A. N.; Sawyer, D. H.; Muller, D. M.

    1983-01-01

    Dichlorosilane (DCS) was used as the feedstock for an advanced decomposition reactor for silicon production. The advanced reactor had a cool bell jar wall temperature, 300 C, when compared to Siemen's reactors previously used for DCS decomposition. Previous reactors had bell jar wall temperatures of approximately 750 C. The cooler wall temperature allows higher DCS flow rates and concentrations. A silicon deposition rate of 2.28 gm/hr-cm was achieved with power consumption of 59 kWh/kg. Interpretation of data suggests that a 2.8 gm/hr-cm deposition rate is possible. Screening of lower cost materials of construction was done as a separate program segment. Stainless Steel (304 and 316), Hastalloy B, Monel 400 and 1010-Carbon Steel were placed individually in an experimental scale reactor. Silicon was deposited from trichlorosilane feedstock. The resultant silicon was analyzed for electrically active and metallic impurities as well as carbon. No material contributed significant amounts of electrically active or metallic impurities, but all contributed carbon.

  17. Neutron measurements with ultra-thin 3D silicon sensors in a radiotherapy treatment room using a Siemens PRIMUS linac

    NASA Astrophysics Data System (ADS)

    Guardiola, C.; Gómez, F.; Fleta, C.; Rodríguez, J.; Quirion, D.; Pellegrini, G.; Lousa, A.; Martínez-de-Olcoz, L.; Pombar, M.; Lozano, M.

    2013-05-01

    The accurate detection and dosimetry of neutrons in mixed and pulsed radiation fields is a demanding instrumental issue with great interest both for the industrial and medical communities. In recent studies of neutron contamination around medical linacs, there is a growing concern about the secondary cancer risk for radiotherapy patients undergoing treatment in photon modalities at energies greater than 6 MV. In this work we present a promising alternative to standard detectors with an active method to measure neutrons around a medical linac using a novel ultra-thin silicon detector with 3D electrodes adapted for neutron detection. The active volume of this planar device is only 10 µm thick, allowing a high gamma rejection, which is necessary to discriminate the neutron signal in the radiotherapy peripheral radiation field with a high gamma background. Different tests have been performed in a clinical facility using a Siemens PRIMUS linac at 6 and 15 MV. The results show a good thermal neutron detection efficiency around 2% and a high gamma rejection factor.

  18. A bidirectional ACR-NEMA interface between the VA's DHCP Integrated Imaging System and the Siemens-Loral PACS.

    PubMed Central

    Kuzmak, P. M.; Dayhoff, R. E.

    1992-01-01

    There is a wide range of requirements for digital hospital imaging systems. Radiology needs very high resolution black and white images. Other diagnostic disciplines need high resolution color imaging capabilities. Images need to be displayed in many locations throughout the hospital. Different imaging systems within a hospital need to cooperate in order to show the whole picture. At the Baltimore VA Medical Center, the DHCP Integrated Imaging System and a commercial Picture Archiving and Communication System (PACS) work in concert to provide a wide-range of departmental and hospital-wide imaging capabilities. An interface between the DHCP and the Siemens-Loral PACS systems enables patient text and image data to be passed between the two systems. The interface uses ACR-NEMA 2.0 Standard messages extended with shadow groups based on draft ACR-NEMA 3.0 prototypes. A Novell file server, accessible to both systems via Ethernet, is used to communicate all the messages. Patient identification information, orders, ADT, procedure status, changes, patient reports, and images are sent between the two systems across the interface. The systems together provide an extensive set of imaging capabilities for both the specialist and the general practitioner. PMID:1482906

  19. Photo neutron dose equivalent rate in 15 MV X-ray beam from a Siemens Primus Linac.

    PubMed

    Ghasemi, A; Pourfallah, T Allahverdi; Akbari, M R; Babapour, H; Shahidi, M

    2015-01-01

    Fast and thermal neutron fluence rates from a 15 MV X-ray beams of a Siemens Primus Linac were measured using bare and moderated BF3 proportional counter inside the treatment room at different locations. Fluence rate values were converted to dose equivalent rate (DER) utilizing conversion factors of American Association of Physicist in Medicine's (AAPM) report number 19. For thermal neutrons, maximum and minimum DERs were 3.46 × 10(-6) (3 m from isocenter in +Y direction, 0 × 0 field size) and 8.36 × 10(-8) Sv/min (in maze, 40 × 40 field size), respectively. For fast neutrons, maximum DERs using 9" and 3" moderators were 1.6 × 10(-5) and 1.74 × 10(-5) Sv/min (2 m from isocenter in +Y direction, 0 × 0 field size), respectively. By changing the field size, the variation in thermal neutron DER was more than the fast neutron DER and the changes in fast neutron DER were not significant in the bunker except inside the radiation field. This study showed that at all points and distances, by decreasing field size of the beam, thermal and fast neutron DER increases and the number of thermal neutrons is more than fast neutrons. PMID:26170555

  20. Comparison of two respiration monitoring systems for 4D imaging with a Siemens CT using a new dynamic breathing phantom

    NASA Astrophysics Data System (ADS)

    Vásquez, A. C.; Runz, A.; Echner, G.; Sroka-Perez, G.; Karger, C. P.

    2012-05-01

    Four-dimensional computed tomography (4D-CT) requires breathing information from the patient, and for this, several systems are available. Testing of these systems, under realistic conditions, requires a phantom with a moving target and an expandable outer contour. An anthropomorphic phantom was developed to simulate patient breathing as well as lung tumor motion. Using the phantom, an optical camera system (GateCT) and a pressure sensor (AZ-733V) were simultaneously operated, and 4D-CTs were reconstructed with a Siemens CT using the provided local-amplitude-based sorting algorithm. The comparison of the tumor trajectories of both systems revealed discrepancies up to 9.7 mm. Breathing signal differences, such as baseline drift, temporal resolution and noise level were shown not to be the reason for this. Instead, the variability of the sampling interval and the accuracy of the sampling rate value written on the header of the GateCT-signal file were identified as the cause. Interpolation to regular sampling intervals and correction of the sampling rate to the actual value removed the observed discrepancies. Consistently, the introduction of sampling interval variability and inaccurate sampling rate values into the header of the AZ-733V file distorted the tumor trajectory for this system. These results underline the importance of testing new equipment thoroughly, especially if components of different manufacturers are combined.

  1. Deposition reactors for solar grade silicon: A comparative thermal analysis of a Siemens reactor and a fluidized bed reactor

    NASA Astrophysics Data System (ADS)

    Ramos, A.; Filtvedt, W. O.; Lindholm, D.; Ramachandran, P. A.; Rodríguez, A.; del Cañizo, C.

    2015-12-01

    Polysilicon production costs contribute approximately to 25-33% of the overall cost of the solar panels and a similar fraction of the total energy invested in their fabrication. Understanding the energy losses and the behaviour of process temperature is an essential requirement as one moves forward to design and build large scale polysilicon manufacturing plants. In this paper we present thermal models for two processes for poly production, viz., the Siemens process using trichlorosilane (TCS) as precursor and the fluid bed process using silane (monosilane, MS). We validate the models with some experimental measurements on prototype laboratory reactors relating the temperature profiles to product quality. A model sensitivity analysis is also performed, and the effects of some key parameters such as reactor wall emissivity and gas distributor temperature, on temperature distribution and product quality are examined. The information presented in this paper is useful for further understanding of the strengths and weaknesses of both deposition technologies, and will help in optimal temperature profiling of these systems aiming at lowering production costs without compromising the solar cell quality.

  2. Synthesis and quality control of fluorodeoxyglucose and performance assessment of Siemens MicroFocus 220 small animal PET scanner

    NASA Astrophysics Data System (ADS)

    Phaterpekar, Siddhesh Nitin

    The scope of this article is to cover the synthesis and quality control procedures involved in production of Fludeoxyglucose (18F--FDG). The article also describes the cyclotron production of 18F radioisotope and gives a brief overview on operations and working of a fixed energy medical cyclotron. The quality control procedures for FDG involve radiochemical and radionuclidic purity tests, pH tests, chemical purity tests, sterility tests, endotoxin tests. Each of these procedures were carried out for multiple batches of FDG with a passing rate of 95% among 20 batches. The article also covers the quality assurance steps for the Siemens MicroPET Focus 220 Scanner using a Jaszczak phantom. We have carried out spatial resolution tests on the scanner, with an average transaxial resolution of 1.775mm with 2-3mm offset. Tests involved detector efficiency, blank scan sinograms and transmission sinograms. A series of radioactivity distribution tests are also carried out on a uniform phantom, denoting the variations in radioactivity and uniformity by using cylindrical ROIs in the transverse region of the final image. The purpose of these quality control tests is to make sure the manufactured FDG is biocompatible with the human body. Quality assurance tests are carried on PET scanners for efficient performance, and to make sure the quality of images acquired is according to the radioactivity distribution in the subject of interest.

  3. Photo neutron dose equivalent rate in 15 MV X-ray beam from a Siemens Primus Linac

    PubMed Central

    Ghasemi, A.; Pourfallah, T. Allahverdi; Akbari, M. R.; Babapour, H.; Shahidi, M.

    2015-01-01

    Fast and thermal neutron fluence rates from a 15 MV X-ray beams of a Siemens Primus Linac were measured using bare and moderated BF3 proportional counter inside the treatment room at different locations. Fluence rate values were converted to dose equivalent rate (DER) utilizing conversion factors of American Association of Physicist in Medicine's (AAPM) report number 19. For thermal neutrons, maximum and minimum DERs were 3.46 × 10-6 (3 m from isocenter in +Y direction, 0 × 0 field size) and 8.36 × 10-8 Sv/min (in maze, 40 × 40 field size), respectively. For fast neutrons, maximum DERs using 9” and 3” moderators were 1.6 × 10-5 and 1.74 × 10-5 Sv/min (2 m from isocenter in +Y direction, 0 × 0 field size), respectively. By changing the field size, the variation in thermal neutron DER was more than the fast neutron DER and the changes in fast neutron DER were not significant in the bunker except inside the radiation field. This study showed that at all points and distances, by decreasing field size of the beam, thermal and fast neutron DER increases and the number of thermal neutrons is more than fast neutrons. PMID:26170555

  4. Reauthorization of the Higher Education Act, TRIO and HEP/CAMP Programs, Volume 5. Hearing before the Subcommittee on Postsecondary Education of the Committee on Education and Labor. House of Representatives, Ninety-Ninth Congress, First Session.

    ERIC Educational Resources Information Center

    Congress of the U.S., Washington, DC. House Committee on Education and Labor.

    A hearing on reauthorization of the Higher Education Act of 1965 is presented that focuses on programs that provide services to students, including the TRIO Programs: Upward Bound, Talent Search, Educational Opportunity Centers, Special Services, and the training authority. It is proposed that secondary and postsecondary schools be helped to…

  5. 26 CFR 1.367(b)-3T - Repatriation of foreign corporate assets in certain nonrecognition transactions (temporary).

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 26 Internal Revenue 4 2014-04-01 2014-04-01 false Repatriation of foreign corporate assets in certain nonrecognition transactions (temporary). 1.367(b)-3T Section 1.367(b)-3T Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY (CONTINUED) INCOME TAX (CONTINUED) INCOME TAXES (CONTINUED) Effects on Corporation §...

  6. 26 CFR 1.67-3T - Allocation of expenses by real estate mortgage investment conduits (temporary).

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... investment conduits (temporary). 1.67-3T Section 1.67-3T Internal Revenue INTERNAL REVENUE SERVICE... mortgage investment conduits (temporary). (a) Allocation of allocable investment expenses—(1) In general. A real estate mortgage investment conduit or REMIC (as defined in section 860D) shall allocate to each...

  7. 26 CFR 1.67-3T - Allocation of expenses by real estate mortgage investment conduits (temporary).

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... investment conduits (temporary). 1.67-3T Section 1.67-3T Internal Revenue INTERNAL REVENUE SERVICE... mortgage investment conduits (temporary). (a) Allocation of allocable investment expenses—(1) In general. A real estate mortgage investment conduit or REMIC (as defined in section 860D) shall allocate to each...

  8. The aporphine alkaloid boldine induces adiponectin expression and regulation in 3T3-L1 cells.

    PubMed

    Yu, Bangning; Cook, Carla; Santanam, Nalini

    2009-10-01

    Adiponectin is an adipokine secreted by differentiated adipocytes. Clinical studies suggest a negative correlation between oxidative stress and adiponectin levels in patients with metabolic syndrome or cardiovascular disease. Natural compounds that can prevent oxidative stress mediated inhibition of adiponectin may be potentially therapeutic. Boldine, an aporphine alkaloid abundant in the medicinal plant Peumus boldus, is a powerful antioxidant. The current study demonstrates the effects of boldine on the expression of adiponectin and its regulators, CCAAT/enhancer binding protein-alpha (C/EBPalpha) and peroxisome proliferator-activated receptor (PPAR)-gamma, in 3T3-L1 cells. Differentiated 3T3-L1 adipocytes were exposed to either hydrogen peroxide (H(2)O(2)) (100 microM) or tumor necrosis factor-alpha (TNFalpha) (1 ng/mL) for 24 hours in the presence or absence of increasing concentrations of boldine (5-100 microM). Quantitative polymerase chain reaction showed that both the oxidants decreased the mRNA levels of adiponectin, PPARgamma, and C/EBPalpha to half of the control levels. Boldine, at all concentrations, counteracted the inhibitory effect of H(2)O(2) or TNFalpha and increased the expression of adiponectin and its regulators. The effect of boldine on adiponectin expression was biphasic, with the lower concentrations (5-25 microM) having a larger inductive effect compared to higher concentrations (50-100 microM). Boldine treatment alone in the absence of H(2)O(2) or TNFalpha was also able to induce adiponectin at the inductive phase of adipogenesis. Peroxisome proliferator response element-luciferase promoter transactivity analysis showed that boldine interacts with the PPAR response element and could potentially modulate PPAR responsive genes. Our results indicate that boldine is able to modulate the expression of adiponectin and its regulators in 3T3-L1 cells and has the potential to be beneficial in obesity-related cardiovascular disease. PMID:19857072

  9. The Aporphine Alkaloid Boldine Induces Adiponectin Expression and Regulation in 3T3-L1 Cells

    PubMed Central

    Yu, Bangning; Cook, Carla

    2009-01-01

    Abstract Adiponectin is an adipokine secreted by differentiated adipocytes. Clinical studies suggest a negative correlation between oxidative stress and adiponectin levels in patients with metabolic syndrome or cardiovascular disease. Natural compounds that can prevent oxidative stress mediated inhibition of adiponectin may be potentially therapeutic. Boldine, an aporphine alkaloid abundant in the medicinal plant Peumus boldus, is a powerful antioxidant. The current study demonstrates the effects of boldine on the expression of adiponectin and its regulators, CCAAT/enhancer binding protein-α (C/EBPα) and peroxisome proliferator-activated receptor (PPAR)-γ, in 3T3-L1 cells. Differentiated 3T3-L1 adipocytes were exposed to either hydrogen peroxide (H2O2) (100 μM) or tumor necrosis factor-α (TNFα) (1 ng/mL) for 24 hours in the presence or absence of increasing concentrations of boldine (5–100 μM). Quantitative polymerase chain reaction showed that both the oxidants decreased the mRNA levels of adiponectin, PPARγ, and C/EBPα to half of the control levels. Boldine, at all concentrations, counteracted the inhibitory effect of H2O2 or TNFα and increased the expression of adiponectin and its regulators. The effect of boldine on adiponectin expression was biphasic, with the lower concentrations (5–25 μM) having a larger inductive effect compared to higher concentrations (50–100 μM). Boldine treatment alone in the absence of H2O2 or TNFα was also able to induce adiponectin at the inductive phase of adipogenesis. Peroxisome proliferator response element-luciferase promoter transactivity analysis showed that boldine interacts with the PPAR response element and could potentially modulate PPAR responsive genes. Our results indicate that boldine is able to modulate the expression of adiponectin and its regulators in 3T3-L1 cells and has the potential to be beneficial in obesity-related cardiovascular disease. PMID:19857072

  10. Nebivolol stimulates mitochondrial biogenesis in 3T3-L1 adipocytes

    SciTech Connect

    Huang, Chenglin; Chen, Dongrui; Xie, Qihai; Yang, Ying; Shen, Weili

    2013-08-16

    Highlights: •Nebivolol may act as a partial agonist of β3-adrenergic receptor (AR). •Nebivolol stimulates mitochondrial DNA replication and protein expression. •Nebivolol promotes mitochondrial synthesis via activation of eNOS by β3-AR. -- Abstract: Nebivolol is a third-generation β-adrenergic receptor (β-AR) blocker with additional beneficial effects, including the improvement of lipid and glucose metabolism in obese individuals. However, the underlying mechanism of nebivolol’s role in regulating the lipid profile remains largely unknown. In this study, we investigated the role of nebivolol in mitochondrial biogenesis in 3T3-L1 adipocytes. Exposure of 3T3-L1 cells to nebivolol for 24 h increased mitochondrial DNA copy number, mitochondrial protein levels and the expression of transcription factors involved in mitochondrial biogenesis, including PPAR-γ coactivator-1α (PGC-1α), Sirtuin 3 (Sirt3), mitochondrial transcription factor A (Tfam) and nuclear related factor 1 (Nrf1). These changes were accompanied by an increase in oxygen consumption and in the expression of genes involved in fatty acid oxidation and antioxidant enzymes in 3T3-L1 adipocytes, including nebivolol-induced endothelial nitric oxide synthase (eNOS), as well as an increase in the formation of cyclic guanosine monophosphate (cGMP). Pretreatment with NG-nitro-L-arginine methyl ester (l-NAME) attenuated nebivolol-induced mitochondrial biogenesis, as did the soluble guanylate cyclase inhibitor, ODQ. Treatment with nebivolol and β3-AR blocker SR59230A markedly attenuated PGC-1α, Sirt3 and manganese superoxide dismutase (MnSOD) protein levels in comparison to treatment with nebivolol alone. These data indicate that the mitochondrial synthesis and metabolism in adipocytes that is promoted by nebivolol is primarily mediated through the eNOS/cGMP-dependent pathway and is initiated by the activation of β3-AR receptors.

  11. Occurrence and control of sporadic proliferation in growth arrested Swiss 3T3 feeder cells.

    PubMed

    Chugh, Rishi Man; Chaturvedi, Madhusudan; Yerneni, Lakshmana Kumar

    2015-01-01

    Growth arrested Swiss mouse embryonic 3T3 cells are used as feeders to support the growth of epidermal keratinocytes and several other target cells. The 3T3 cells have been extensively subcultured owing to their popularity and wide distribution in the world and, as a consequence selective inclusion of variants is a strong possibility in them. Inadvertently selected variants expressing innate resistance to mitomycin C may continue to proliferate even after treatment with such growth arresting agents. The failure of growth arrest can lead to a serious risk of proliferative feeder contamination in target cell cultures. In this study, we passaged Swiss 3T3 cells (CCL-92, ATCC) by different seeding densities and incubation periods. We tested the resultant cultures for differences in anchorage-independent growth, resumption of proliferation after mitomycin C treatment and occurrence of proliferative feeder contaminants in an epidermal keratinocyte co-culture system. The study revealed subculture dependent differential responses. The cultures of a particular subculture procedure displayed unique cell size distribution and disintegrated completely in 6 weeks following mitomycin C treatment, but their repeated subculture resulted in feeder regrowth as late as 11 weeks after the growth arrest. In contrast, mitomycin C failed to inhibit cell proliferation in cultures of the other subculture schemes and also in a clone that was established from a transformation focus of super-confluent culture. The resultant proliferative feeder cells contaminated the keratinocyte cultures. The anchorage-independent growth appeared in late passages as compared with the expression of mitomycin C resistance in earlier passages. The feeder regrowth was prevented by identifying a safe subculture protocol that discouraged the inclusion of resistant variants. We advocate routine anchorage-independent growth assay and absolute confirmation of feeder disintegration to qualify feeder batches and

  12. Lysophosphatidic acid induces chemotaxis in MC3T3-E1 osteoblastic cells.

    PubMed

    Masiello, Lisa M; Fotos, Joseph S; Galileo, Deni S; Karin, Norman J

    2006-07-01

    Lysophosphatidic acid (LPA) is a bioactive lipid that has pleiotropic effects on a variety of cell types and enhances the migration of endothelial and cancer cells, but it is not known if this lipid can alter osteoblast motility. We performed transwell migration assays using MC3T3-E1 osteoblastic cells and found LPA to be a potent chemotactic agent. Quantitative time-lapse video analysis of osteoblast migration after wounds were introduced into cell monolayers indicated that LPA stimulated both migration velocity and the average migration distance per cell. LPA also elicited substantial changes in cell shape and actin cytoskeletal structure; lipid-treated cells contained fewer stress fibers and displayed long membrane processes that were enriched in F-actin. Quantitative RT-PCR analysis showed that MC3T3-E1 cells express all four known LPA-specific G-protein-coupled receptors (LPA1-LPA4) with a relative mRNA abundance of LPA1>LPA4>LPA2>LPA3. LPA-induced changes in osteoblast motility and morphology were antagonized by both pertussis toxin and Ki16425, a subtype-specific blocker of LPA1 and LPA3 receptor function. Cell migration in many cell types is linked to changes in intracellular Ca2+. Ki16425 also inhibited LPA-induced Ca2+ signaling in a dose-dependent manner, suggesting a link between LPA-induced Ca2+ transients and osteoblast chemotaxis. Our data show that LPA stimulates MC3T3-E1 osteoblast motility via a mechanism that is linked primarily to the G-protein-coupled receptor LPA1. PMID:16487757

  13. Lysophosphatidic acid induces chemotaxis in MC3T3-E1 osteoblastic cells

    SciTech Connect

    Masiello, Lisa M.; Fotos, Joseph S.; Galileo, Deni S.; Karin, Norm J.

    2006-07-01

    Lysophosphatidic acid (LPA) is a bioactive lipid that has pleiotropic effects on a variety of cell types and enhances the migration of endothelial and cancer cells, but it is not known if this lipid can alter osteoblast motility. We performed transwell migration assays using MC3T3-E1 osteoblastic cells and found LPA to be a potent chemotactic agent. Quantitative time-lapse video analysis of osteoblast migration after wounds were introduced into cell monolayers indicated that LPA stimulated both migration velocity and the average migration distance per cell. LPA also elicited substantial changes in cell shape and actin cytoskeletal structure; lipid-treated cells contained fewer stress fibers and displayed long membrane processes that were enriched in F-actin. Quantitative RT-PCR analysis showed that MC3T3-E1 cells express all four known LPA-specific G protein-coupled receptors (LPA1-LPA4) with a relative mRNA abundance of LPA1 > LPA4 > LPA2 >> LPA3. LPA-induced changes in osteoblast motility and morphology were antagonized by both pertussis toxin and Ki16425, a subtype-specific blocker of LPA1 and LPA3 receptor function. Cell migration in many cell types is linked to changes in intracellular Ca2+. Ki16425 also inhibited LPA-induced Ca2+ signaling in a dose-dependent manner, suggesting a link between LPA-induced Ca2+ transients and osteoblast chemotaxis. Our data show that LPA stimulates MC3T3-E1 osteoblast motility via a mechanism that is linked primarily to the G protein-coupled receptor LPA1.

  14. 3T diffusion-weighted MRI of the thyroid gland with reduced distortion: preliminary results

    PubMed Central

    Nagala, S; Priest, A N; McLean, M A; Jani, P; Graves, M J

    2013-01-01

    Objective: Single-shot diffusion-weighted (DW) echo planar imaging (EPI), which is commonly used for imaging the thyroid, is characterised by severe blurring and distortion. The objectives of this work were: 1, to show that a reduced-field of view (r-FOV) DW EPI technique can improve image quality; and 2, to investigate the effect of different reconstruction strategies on the resulting apparent diffusion coefficients (ADCs). Methods: We implemented a single-shot, r-FOV DW EPI technique with a two-dimensional radiofrequency excitation pulse for DW imaging of the thyroid at 3T. Images were reconstructed using root sum of squares (SOS) and an optimal-B1 reconstruction (OBR). Phantom and in vivo experiments were performed to compare r-FOV and conventional full-FOV DW EPI with root SOS and OBR. Results: r-FOV with OBR substantially improved image quality at 3T. In phantoms, r-FOV gave more accurate ADCs than full-FOV. In vivo r-FOV always gave lower ADC values with respect to the full-FOV technique irrespective of the reconstruction used and whether only two or multiple b-values were used to compute the ADCs. Conclusion: r-FOV DW EPI can reduce image blurring and distortion at the expense of a low signal-to-noise ratio. OBR is a promising reconstruction technique for accurate ADC measurements in lower signal-to-noise ratio regimes, although further studies are needed to characterise its performance. Advances in knowledge: DW imaging of the thyroid at 3T could potentially benefit from r-FOV acquisition strategies, such as the r-FOV DW EPI technique proposed in this paper. PMID:23770539

  15. Changes in laser-induced fluorescence responses of 3T3 fibroblasts to repetitive thermal stress

    NASA Astrophysics Data System (ADS)

    Beuthan, J.; Dressler, C.; Zabarylo, U.; Minet, O.

    2009-04-01

    The combined experimental use of laser-induced autofluorescence of cellular metabolites and methodological fundamentals of systems biology will provide access to biological thermal stress analysis on a sub cellular level. A test setup incorporating a pulsed nitrogen laser was realized with which autofluorescence of the coenzyme NADH could be measured in living 3T3 cells. The cells were subjected to different temperature stress at repetitive time intervals. When subjected to a simple mathematical analysis, the NADH concentration change measured through autofluorescence in biological cells exhibited approximate concentration-equivalent balance curves. These results add up to the fundamental know-how about the dosimetry of thermally therapeutic methods.

  16. NYU3T: teaching, technology, teamwork: a model for interprofessional education scalability and sustainability.

    PubMed

    Djukic, Maja; Fulmer, Terry; Adams, Jennifer G; Lee, Sabrina; Triola, Marc M

    2012-09-01

    Interprofessional education is a critical precursor to effective teamwork and the collaboration of health care professionals in clinical settings. Numerous barriers have been identified that preclude scalable and sustainable interprofessional education (IPE) efforts. This article describes NYU3T: Teaching, Technology, Teamwork, a model that uses novel technologies such as Web-based learning, virtual patients, and high-fidelity simulation to overcome some of the common barriers and drive implementation of evidence-based teamwork curricula. It outlines the program's curricular components, implementation strategy, evaluation methods, and lessons learned from the first year of delivery and describes implications for future large-scale IPE initiatives. PMID:22920424

  17. Vinculin expression in MC3T3-E1 cells in response to mechanical stimulus

    PubMed Central

    Cora-Cruz, J.J.; Diffoot-Carlo, N.; Sundaram, P.A.

    2015-01-01

    Loading frequency is known to influence the expression of the focal adhesions of the adherent cells. A small cyclical tensile force was transmitted to mouse pre-osteoblast MC3T3-E1 cells through PDMS substrates of varying stiffness. Changes in cell behavior with respect to proliferation and characteristics of focal adhesions were quantified through immunofluorescence labeling of vinculin. Amount of inactive vinculin was higher on substrates subjected to cyclic stimulation when compared with the results of the static substrates, whereas the number and area of focal adhesion points underwent a reduction. Inactive vinculin appears as a cloud in the cytoplasm in the vicinity of the nucleus. PMID:26858974

  18. Improved SNR of magnetic resonance microimaging using a cooled resonance circuit at 0.3T.

    PubMed

    Nakano, Hiroshi; Nakai, Toshiharu

    2011-01-01

    Because it is noninvasive, magnetic resonance microimaging (MRMI) can be used for 3-dimensional measurement of living tissues for cell engineering. Thermal noise in the resonance circuit of the radiofrequency (RF) system of the MRMI cannot become ignored as the signal diminishes in accordance with decreasing sample size, and cooling the RF coil of the receiver circuit can effectively reduce thermal noise. We used a low temperature normal conductor circuit to reduce noise and confirmed improved signal-to-noise ratio for a conventional microimaging system at low B(0) field (0.3T) with low cost. PMID:22214912

  19. The 3' end CCA of mature tRNA is an antideterminant for eukaryotic 3'-tRNase.

    PubMed Central

    Mohan, A; Whyte, S; Wang, X; Nashimoto, M; Levinger, L

    1999-01-01

    Cytoplasmic tRNAs undergo posttranscriptional 5' and 3' end processing in the eukaryotic nucleus, and CCA (which forms the mature 3' end of all tRNAs) must be added by tRNA nucleotidyl transferase before tRNA can be aminoacylated and utilized in translation. Eukaryotic 3'-tRNase can endonucleolytically remove a 3' end trailer by cleaving on the 3' side of the discriminator base (the unpaired nucleotide 3' of the last base pair of the acceptor stem). This reaction proceeds despite a wide range in length and sequence of the 3' end trailer, except that mature tRNA containing the 3' terminal CCA is not a substrate for mouse 3'-tRNase (Nashimoto, 1997, Nucleic Acids Res 25:1148-1154). Herein, we extend this result with Drosophila and pig 3'-tRNase, using Drosophila melanogaster tRNAHis as substrate. Mature tRNA is thus prevented from recycling through 3' end processing. We also tested a series of tRNAs ending at the discriminator base (-), with one C added (+C), two Cs added (+CC), and CCA added (+CCA) as 3'-tRNase inhibitors. Inhibition was competitive with both Drosophila and pig 3'-tRNase. The product of the 3'-tRNase reaction (-) is a good 3'-tRNase inhibitor, with a KI approximately two times KM for the normal 3'-tRNase substrate. KI increases with each nucleotide added beyond the discriminator base, until when tRNA+CCA is used as inhibitor, KI is approximately forty times the substrate KM. The 3'-tRNase can thus remain free to process precursors with 3' end trailers because it is barely inhibited by tRNA+CCA, ensuring that tRNA can progress to aminoacylation. The active site of 3'-tRNase may have evolved to make an especially poor fit with tRNA+CCA. PMID:10024176

  20. Measurement and control system for cryogenic helium gas bearing turbo-expander experimental platform based on Siemens PLC S7-300

    NASA Astrophysics Data System (ADS)

    Li, J.; Xiong, L. Y.; Peng, N.; Dong, B.; Wang, P.; Liu, L. Q.

    2014-01-01

    An experimental platform for cryogenic Helium gas bearing turbo-expanders is established at the Technical Institute of Physics and Chemistry, Chinese Academy of Sciences. This turbo-expander experimental platform is designed for performance testing and experimental research on Helium turbo-expanders with different sizes from the liquid hydrogen temperature to the room temperature region. A measurement and control system based on Siemens PLC S7-300 for this turbo-expander experimental platform is developed. Proper sensors are selected to measure such parameters as temperature, pressure, rotation speed and air flow rate. All the collected data to be processed are transformed and transmitted to S7-300 CPU. Siemens S7-300 series PLC CPU315-2PN/DP is as master station and two sets of ET200M DP remote expand I/O is as slave station. Profibus-DP field communication is established between master station and slave stations. The upper computer Human Machine Interface (HMI) is compiled using Siemens configuration software WinCC V6.2. The upper computer communicates with PLC by means of industrial Ethernet. Centralized monitoring and distributed control is achieved. Experimental results show that this measurement and control system has fulfilled the test requirement for the turbo-expander experimental platform.

  1. Development of a Measurement and Control System for a 40l/h Helium Liquefier based on Siemens PLC S7-300

    NASA Astrophysics Data System (ADS)

    Li, J.; Liu, L. Q.; Xu, X. D.; Liu, T.; Li, Q.; Hu, Z. J.; Wang, B. M.; Xiong, L. Y.; Dong, B.; Yan, T.

    A 40l/h Helium Liquefier has been commissioned by the Technical Institute of Physics and Chemistry, Chinese Academy of Sciences. A measurement and control system based on Siemens PLC S7-300 for this Helium Liquefier is developed. Proper sensors are selected, for example, three types of transmitters are adopted respectively according to detailed temperature measurement requirements. Siemens S7-300 PLC CPU315-2PN/DP operates as a master station and three sets of ET200 M DP remote expand I/O operate asslave stations. Profibus-DP field communication is used between the master station and the slave stations. The upper computer HMI(Human Machine Interface) is compiled using Siemens configuration software WinCC V7.0. The upper computer communicates with PLC by means of industrial Ethernet. A specific control logic for this Helium Liquefier is developed. The control of the suction and discharge pressures of the compressor and the control of the turbo-expanders loop are being discussed in this paper. Following the commissioning phase, the outlet temperature of the second stage turbine has reached 8.6K and the temperature before the throttle valve has reached 13.1K.

  2. Measurement and control system for cryogenic helium gas bearing turbo-expander experimental platform based on Siemens PLC S7-300

    SciTech Connect

    Li, J.; Xiong, L. Y.; Peng, N.; Dong, B.; Liu, L. Q.; Wang, P.

    2014-01-29

    An experimental platform for cryogenic Helium gas bearing turbo-expanders is established at the Technical Institute of Physics and Chemistry, Chinese Academy of Sciences. This turbo-expander experimental platform is designed for performance testing and experimental research on Helium turbo-expanders with different sizes from the liquid hydrogen temperature to the room temperature region. A measurement and control system based on Siemens PLC S7-300 for this turbo-expander experimental platform is developed. Proper sensors are selected to measure such parameters as temperature, pressure, rotation speed and air flow rate. All the collected data to be processed are transformed and transmitted to S7-300 CPU. Siemens S7-300 series PLC CPU315-2PN/DP is as master station and two sets of ET200M DP remote expand I/O is as slave station. Profibus-DP field communication is established between master station and slave stations. The upper computer Human Machine Interface (HMI) is compiled using Siemens configuration software WinCC V6.2. The upper computer communicates with PLC by means of industrial Ethernet. Centralized monitoring and distributed control is achieved. Experimental results show that this measurement and control system has fulfilled the test requirement for the turbo-expander experimental platform.

  3. Hippo-independent activation of YAP by the GNAQ uveal melanoma oncogene through a trio-regulated rho GTPase signaling circuitry.

    PubMed

    Feng, Xiaodong; Degese, Maria Sol; Iglesias-Bartolome, Ramiro; Vaque, Jose P; Molinolo, Alfredo A; Rodrigues, Murilo; Zaidi, M Raza; Ksander, Bruce R; Merlino, Glenn; Sodhi, Akrit; Chen, Qianming; Gutkind, J Silvio

    2014-06-16

    Mutually exclusive activating mutations in the GNAQ and GNA11 oncogenes, encoding heterotrimeric Gαq family members, have been identified in ∼ 83% and ∼ 6% of uveal and skin melanomas, respectively. However, the molecular events underlying these GNAQ-driven malignancies are not yet defined, thus limiting the ability to develop cancer-targeted therapies. Here, we focused on the transcriptional coactivator YAP, a critical component of the Hippo signaling pathway that controls organ size. We found that Gαq stimulates YAP through a Trio-Rho/Rac signaling circuitry promoting actin polymerization, independently of phospholipase Cβ and the canonical Hippo pathway. Furthermore, we show that Gαq promotes the YAP-dependent growth of uveal melanoma cells, thereby identifying YAP as a suitable therapeutic target in uveal melanoma, a GNAQ/GNA11-initiated human malignancy. PMID:24882515

  4. Effect of pycnogenol on glucose transport in mature 3T3-L1 adipocytes.

    PubMed

    Lee, Hee-Hyun; Kim, Kui-Jin; Lee, Ok-Hwan; Lee, Boo-Yong

    2010-08-01

    Pycnogenol, a procyanidins-enriched extract of Pinus maritima bark, possesses antidiabetic properties, which improves the altered parameters of glucose metabolism that are associated with type 2 diabetes mellitus (T2DM). Since the insulin-stimulated antidiabetic activities of natural bioactive compounds are mediated by GLUT4 via the phosphatidylinositol-3-kinase (PI3K) and/or p38 mitogen activated protein kinase (p38-MAPK) pathway, the effects of pycnogenol were examined on the molecular mechanism of glucose uptake by the glucose transport system. 3T3-L1 adipocytes were treated with various concentrations of pycnogenol, and glucose uptake was examined using a non-radioisotope enzymatic assay and by molecular events associated with the glucose transport system using semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR). The results show that pycnogenol increased glucose uptake in fully differentiated 3T3-L1 adipocytes and increased the relative abundance of both GLUT4 and Akt mRNAs through the PI3K pathway in a dose dependent manner. Furthermore, pycnogenol restored the PI3K antagonist-induced inhibition of glucose uptake in the presence of wartmannin, an inhibitor of the PI3K. Overall, these results indicate that pycnogenol may stimulate glucose uptake via the PI3K dependent tyrosine kinase pathways involving Akt. Further the results suggest that pycnogenol might be useful in maintaining blood glucose control. PMID:20658573

  5. Bombesin stimulation of DNA synthesis and cell division in cultures of Swiss 3T3 cells.

    PubMed Central

    Rozengurt, E; Sinnett-Smith, J

    1983-01-01

    Bombesin is shown to be a potent mitogen for Swiss 3T3 cells. At nanomolar concentrations the peptide markedly enhances the ability of fresh serum to stimulate DNA synthesis in confluent and quiescent cultures of these cells. In the presence of a low concentration (3.5%) of serum, bombesin stimulates 3T3 cell proliferation. In serum-free medium, bombesin induces DNA synthesis in the absence of any other added growth factor; half-maximal effect is obtained at 1 nM. The mitogenic effect of bombesin is dependent on dose and time, is mimicked by litorin, and is markedly potentiated by insulin, colchicine, platelet-derived growth factor, and fibroblast-derived growth factor. These mitogens increase the maximal response elicited by bombesin and decrease the bombesin concentration required to produce half-maximal effect (from 1 nM to 0.3 nM). In contrast, vasopressin, phorbol esters, or cAMP increasing agents fail to enhance the maximal level of DNA synthesis induced by bombesin. Bombesin and litorin may provide useful model peptides for studies on the mechanism(s) by which extracellular ligands control cell proliferation. PMID:6344074

  6. Poly(L-lactide) crystallization topography directs MC3T3-E1 cells response.

    PubMed

    Li, Wenqiang; Lu, Lu; Jiao, Yanpeng; Zhang, Chaowen; Zhou, Changren

    2016-09-01

    Biomaterial surface topography significantly influences cellular form and function. Using poly(L-lactic acid) films with normal spherulites, banded spherulites, and amorphous surfaces as model substrates, we conducted a systematic assessment of the role for polymer crystallization induced surface morphologies on cell growth and contact guidance. Microscopy and image analysis showed that the MC3T3-E1 cells spread out in a random fashion on the amorphous substrate. At 24 h post-seeding, MC3T3-E1 cells on both types of spherulite surfaces were elongated and aligned along the spherulite radius direction. For the banded spherulite surface with radial stripes and coupling annular grooves, the cell orientation and cell nuclear localization were related to the grooves structure. With increasing time, this orientation preference was weaker. These results demonstrate that the patterning of polymer crystallization structure provide important signals for guiding cells to exhibit characteristic orientation and morphology especially in the early stages of regeneration. PMID:27376548

  7. Persistent induction of cyclooxygenase in p60 sup v-src -transformed 3T3 fibroblasts

    SciTech Connect

    Han, Jiawen; Sadowski, H.; Young, D.A.; Macara, I.G. )

    1990-05-01

    A BALB/c 3T3 cell line infected with the temperature-sensitive Rous sarcoma virus strain LA90 has been used to investigate early, p60{sup v-src}-dependent changes in gene expression (protein synthesis). Giant two-dimensional electrophoresis, which can resolve >3,000 polypeptides from ({sup 35}S)methionine-labeled cell lysates, was used to detect the induction of a p72-74 (72-74 kDa) doublet (pI 7.5) after activation of p60{sup v-src} at 35{degree}C. Antiserum against cyclooxygenase (prostaglandin synthase or prostaglandin endoperoxide synthase) specifically immunoprecipitated the p72-74 doublet. The p72-74 doublet was also induced by platelet-derived growth factor and by phorbol 12-myristate 13-acetate and was elevated in an NIH 3T3 cell line transformed by wild-type src. Activation of p60{sup v-src} caused a persistant increase in p72-74, whereas the effect of the growth factor was transient. These dissimilar kinetics of induction were paralleled by changes in cyclooxygenase activity. Although induction of this enzyme may not be directly involved in transformation, the data support the view that oncogenic transformation may result, not from expression of transformation-specific genes, but from persistent changes in the expression of genes normally induced only transiently during passage from the G{sub 0} stage of the cell cycle.

  8. RA induces the neural-like cells generated from epigenetic modified NIH/3T3 cells.

    PubMed

    Zhang, Xi-Mei; Li, Qiu-Ming; Su, Dong-Ju; Wang, Ning; Shan, Zhi-Yan; Jin, Lian-Hong; Lei, Lei

    2010-03-01

    Recently, differentiated somatic cells had been reprogrammed to pluripotential state in vitro, and various tissue cells had been elicited from those cells. Epigenetic modifications allow differentiated cells to perpetuate the molecular memory needed for the cells to retain their identity. DNA methylation and histone deacetylation are important patterns involved in epigenetic modification, which take critical roles in regulating DNA expression. In this study, we dedifferentiated NIH/3T3 fibroblasts by 5-aza-2-deoxycytidine (5-aza-dC) and Trichstatin A (TSA) combination, and detected gene expression pattern, DNA methylation level, and differentiation potential of reprogrammed cells. As the results, embryonic marker Sox2, klf4, c-Myc and Oct4 were expressed in reprogrammed NIH/3T3 fibroblasts. Total DNA methylation level was significant decreased after the treatment. Moreover, exposure of the reprogrammed cells to all trans-retinoic acid (RA) medium elicited the generation of neuronal class IIIbeta-tubulin-positive, neuron-specific enolase (NSE)-positive, nestin-positive, and neurofilament light chain (NF-L)-positive neural-like cells. PMID:19263240

  9. The effect of 3 T MRI on microleakage of amalgam restorations

    PubMed Central

    Yilmaz, S; Misirlioğlu, M

    2013-01-01

    Objectives: To evaluate the effects of 3 T magnetic field on microleakage of amalgam restorations containing three different types of silver (Ag). Methods: 60 extracted teeth were restored with three different types of amalgam filling materials. Restored teeth were sectioned mesiodistally and divided into experimental and control groups. Experimental groups were exposed to a magnetic field of 3 T for 20 min. All samples were plunged into 2% basic fuchsin solution and examined under a digital microscope by three different observers with regard to microleakage. Results: Statistical analysis showed significant differences in microleakage between the groups exposed to MRI and controls, whereas differences in microleakage between amalgam types were insignificant. Conclusions: The primary risk of MRI systems arises from the effects of its strong magnetic field on objects containing ferromagnetic materials. An MRI of 1.5 T is known to be safe for amalgam restorations. However, our research indicates that MRI is not completely devoid of any effects on amalgam restorations. PMID:23674614

  10. An optically powered CMOS tracking system for 3 T magnetic resonance environment.

    PubMed

    Sarioglu, Baykal; Tumer, Murat; Cindemir, Umut; Camli, Berk; Dundar, Gunhan; Ozturk, Cengizhan; Yalcinkaya, Arda D

    2015-02-01

    In this work, a fully optical Complementary Metal Oxide Semiconductor (CMOS) based catheter tracking system designed for 3 T Magnetic Resonance Imaging (MRI) environment is presented. The system aims to solve the Radio Frequency (RF) induced heating problem present in conventional wired catheter tracking systems used in MRI. It is based on an integrated circuit, consisting of a receiver and an optical power supply unit. The optical power supply unit includes a single on-chip photodiode and a DC-DC converter that boosts the low photodiode voltage output to voltages greater than 1.5 V. Through an optically driven switch, the accumulated charge on an a storage capacitor is transferred to the rest of the system. This operation is novel in the way that it is fully optical and the switch control is done through modulation of the applied light. An on-chip local oscillator signal for the receiver is avoided by application of an RF signal that is generated by the MRI machine at the receiving period. The signals received by a micro-coil antenna are processed by the on-chip direct conversion receiver. The processed signal is then transferred, also optically, to the outside world for tracking purposes. The frequency encoding method is used for MRI tracking. Operation with various levels of external optical power does not generate noticeble temperature increase in the system. The overall system is successfully tested in a 3 T MRI machine to demonstrate its full operation. PMID:24893369

  11. Polyamine metabolism is involved in adipogenesis of 3T3-L1 cells.

    PubMed

    Ishii, Ikumi; Ikeguchi, Yoshihiko; Mano, Hiroshi; Wada, Masahiro; Pegg, Anthony E; Shirahata, Akira

    2012-02-01

    Polyamines spermidine and spermine are known to be required for mammalian cell proliferation and for embryonic development. Alpha-difluoromethylornithine (DFMO), an inhibitor of ornithine decarboxylase (ODC) a limiting enzyme of polyamine biosynthesis, depleted the cellular polyamines and prevented triglyceride accumulation and differentiation in 3T3-L1 cells. In this study, to explore the function of polyamines in adipogenesis, we examined the effect of polyamine biosynthesis inhibitors on adipocyte differentiation and lipid accumulation of 3T3-L1 cells. The spermidine synthase inhibitor trans-4-methylcyclohexylamine (MCHA) increased spermine/spermidine ratios, whereas the spermine synthase inhibitor N-(3-aminopropyl)-cyclohexylamine (APCHA) decreased the ratios in the cells. MCHA was found to decrease lipid accumulation and GPDH activity during differentiation, while APCHA increased lipid accumulation and GPDH activity indicating the enhancement of differentiation. The polyamine-acetylating enzyme, spermidine/spermine N(1)-acetyltransferase (SSAT) activity was increased within a few hours after stimulus for differentiation, and was found to be elevated by APCHA. In mature adipocytes APCHA decreased lipid accumulation while MCHA had the opposite effect. An acetylpolyamine oxidase and spermine oxidase inhibitor MDL72527 or an antioxidant N-acetylcysteine prevented the promoting effect of APCHA on adipogenesis. These results suggest that not only spermine/spermidine ratios but also polyamine catabolic enzyme activity may contribute to adipogenesis. PMID:21809076

  12. Oxidative changes and apoptosis induced by 1800-MHz electromagnetic radiation in NIH/3T3 cells.

    PubMed

    Hou, Qingxia; Wang, Minglian; Wu, Shuicai; Ma, Xuemei; An, Guangzhou; Liu, Huan; Xie, Fei

    2015-03-01

    To investigate the potential adverse effects of mobile phone radiation, we studied reactive oxygen species (ROS), DNA damage and apoptosis in mouse embryonic fibroblasts (NIH/3T3) after intermittent exposure (5 min on/10 min off, for various durations from 0.5 to 8 h) to an 1800-MHz GSM-talk mode electromagnetic radiation (EMR) at an average specific absorption rate of 2 W/kg. A 2',7'-dichlorofluorescin diacetate fluorescence probe was used to detect intracellular ROS levels, immunofluorescence was used to detect γH2AX foci as a marker for DNA damage, and flow cytometry was used to measure apoptosis. Our results showed a significant increase in intracellular ROS levels after EMR exposure and it reached the highest level at an exposure time of 1 h (p < 0.05) followed by a slight decrease when the exposure continued for as long as 8 h. No significant effect on the number of γH2AX was detected after EMR exposure. The percentage of late-apoptotic cells in the EMR-exposed group was significantly higher than that in the sham-exposed groups (p < 0.05). These results indicate that an 1800-MHz EMR enhances ROS formation and promotes apoptosis in NIH/3T3 cells. PMID:24665905

  13. High-affinity receptors for peptides of the bombesin family in Swiss 3T3 cells

    SciTech Connect

    Zachary, I.; Rozengurt, E.

    1985-11-01

    Gastrin-releasing peptide (GRP) labeled with /sup 125/I at tyrosine-15 (/sup 125/I-GRP) binds to intact quiescent Swiss 3T3 cells in a specific and saturable manner. Scatchard analysis indicates the presence of a single class of high-affinity binding sites of Kd = 0.5 X 10(-9) M and a value for the number of sites per cell of about 100,000. /sup 125/I-GRP binding was not inhibited by other mitogens for these cells, and cell lines that are mitogenically unresponsive to GRP do not exhibit specific GRP binding. Structure-activity relationships show a close parallel between the ability of a range of GRP-related peptides to both inhibit GRP binding and to stimulate mitogenesis. Further, GRP binding is selectively blocked in a competitive fashion by a novel bombesin antagonist, (D-Arg1, D-Pro2, D-Trp7,9, Leu11) substance P. In addition, this compound selectively inhibits GRP and bombesin-induced mitogenesis. These results demonstrate that the mitogenic response of Swiss 3T3 cells to peptides of the bombesin family is mediated by a class of receptors distinct from those of other mitogens for these cells.

  14. Conventional kinesin KIF5B mediates adiponectin secretion in 3T3-L1 adipocytes.

    PubMed

    Cui, Ju; Pang, Jing; Lin, Ya-Jun; Jiang, Ping; Gong, Huan; Wang, Zai; Li, Jian; Cai, Jian-Ping; Huang, Jian-Dong; Zhang, Tie-Mei

    2016-08-01

    Insulin stimulates adiponectin secretion and glucose transporter type 4 (GLUT4) translocation in adipocyte to regulate metabolism homeostasis. Similar to GLUT4 translocation, intracellular trafficking and release of adiponectin in adipocytes relies on the trans-Golgi network and endosomal system. Recent studies show that the heavy chain of conventional kinesin (KIF5B) mediates GLUT4 translocation in murine 3T3-L1 adipocytes, however, the motor machinery involved in mediating intracellular trafficking and release of adiponectin is unknown. Here, we examined the role of KIF5B in the regulation of adiponectin secretion. The KIF5B level was up-regulated during 3T3-L1 adipogenesis. This increase in cytosolic KIF5B was synchronized with the induction of adiponectin. Endogenous KIF5B and adiponectin were partially colocalized at the peri-nuclear and cytosolic regions. In addition, adiponectin-containing vesicles were co-immunoprecipitated with KIF5B. Knockdown of KIF5B resulted in a marked inhibition of adiponectin secretion and overexpression of KIF5B enhanced adiponectin release, whereas leptin secretion was not affected by changes in KIF5B expression. These data suggest that the secretion of adiponectin, but not leptin, is dependent on functional KIF5B. PMID:27264953

  15. Bombesin, vasopressin, and endothelin rapidly stimulate tyrosine phosphorylation in intact Swiss 3T3 cells

    SciTech Connect

    Zachary, I.; Gil, J.; Lehmann, W.; Sinnett-Smith, J.; Rozengurt, E. )

    1991-06-01

    The mitogenic neuropeptides bombesin and vasopressin markedly increased tyrosine and serine phosphorylation of multiple substrates in quiescent Swiss 3T3 fibroblasts, including two major bands of M{sub r} 90,000 and 115,000. Tyrosine phosphorylation of these proteins was increased as judged by immunoprecipitation of {sup 32}P{sub i}-labeled cells and immunoblotting of unlabeled cells with monoclonal antiphosphotyrosine antibodies, elution with phenyl phosphate, and phospho amino acid analysis. Phosphotyrosyl proteins generated by bombesin and vasopressin did not correspond either by apparent molecular weight or by immunological and biochemical criteria to several known tyrosine kinase substrates, including phospholipase C{sub {gamma}}, the microtubule-associated protein 2 kinase, GTPase-activating protein, or phosphatidylinositol kinase. The effect was rapid (within seconds), concentration dependent, and inhibited by specific receptor antagonists for both bombesin and vasopressin. The endothelin-related peptide, vasoactive intestinal contractor, also elicited a rapid and concentration-dependent tyrosine/serine phosphorylation of a similar set of substrates. These results demonstrate that neuropeptides, acting through receptors linked to GTP-binding proteins, stimulate tyrosine phosphorylation of a common set of substrates in quiescent Swiss 3T3 cells and suggest the existence of an additional signal transduction pathway in neuropeptide-induced mitogenesis.

  16. The Depletion of Nuclear Glutathione Impairs Cell Proliferation in 3t3 Fibroblasts

    PubMed Central

    Markovic, Jelena; Mora, Nancy J.; Broseta, Ana M.; Gimeno, Amparo; de-la-Concepción, Noelia; Viña, José; Pallardó, Federico V.

    2009-01-01

    Background Glutathione is considered essential for survival in mammalian cells and yeast but not in prokaryotic cells. The presence of a nuclear pool of glutathione has been demonstrated but its role in cellular proliferation and differentiation is still a matter of debate. Principal Findings We have studied proliferation of 3T3 fibroblasts for a period of 5 days. Cells were treated with two well known depleting agents, diethyl maleate (DEM) and buthionine sulfoximine (BSO), and the cellular and nuclear glutathione levels were assessed by analytical and confocal microscopic techniques, respectively. Both agents decreased total cellular glutathione although depletion by BSO was more sustained. However, the nuclear glutathione pool resisted depletion by BSO but not with DEM. Interestingly, cell proliferation was impaired by DEM, but not by BSO. Treating the cells simultaneously with DEM and with glutathione ethyl ester to restore intracellular GSH levels completely prevented the effects of DEM on cell proliferation. Conclusions Our results demonstrate the importance of nuclear glutathione in the control of cell proliferation in 3T3 fibroblasts and suggest that a reduced nuclear environment is necessary for cells to progress in the cell cycle. PMID:19641610

  17. Stereo ENA Imaging of the Ring Current and Multi-point Measurements of Suprathermal Particles and Magnetic Fields by TRIO-CINEMA

    NASA Astrophysics Data System (ADS)

    Lin, R. P.; Sample, J. G.; Immel, T. J.; Lee, D.; Horbury, T. S.; Jin, H.; SEON, J.; Wang, L.; Roelof, E. C.; Lee, E.; Parks, G. K.; Vo, H.

    2012-12-01

    The TRIO (Triplet Ionospheric Observatory) - CINEMA (Cubesat for Ions, Neutrals, Electrons, & Magnetic fields) mission consists of three identical 3-u cubesats to provide high sensitivity, high cadence, stereo measurements of Energetic Neutral Atoms (ENAs) from the Earth's ring current with ~1 keV FWHM energy resolution from ~4 to ~200 keV, as well as multi-point in situ measurements of magnetic fields and suprathermal electrons (~2 -200 keV) and ions (~ 4 -200 keV) in the auroral and ring current precipitation regions in low Earth orbit (LEO). A new Suprathermal Electron, Ion, Neutral (STEIN) instrument, using a 32-pixel silicon semiconductor detector with an electrostatic deflection system to separate ENAs from ions and from electrons below 30 keV, will sweep over most of the sky every 15 s as the spacecraft spins at 4 rpm. In addition, inboard and outboard (on an extendable 1m boom) miniature magnetoresistive sensor magnetometers will provide high cadence 3-axis magnetic field measurements. An S-band transmitter will be used to provide ~8 kbps orbit-average data downlink to the ~11m diameter antenna of the Berkeley Ground Station.The first CINEMA (funded by NSF) is scheduled for launch on August 14, 2012 into a 65 deg. inclination LEO. Two more identical CINEMAs are being developed by Kyung Hee University (KHU) in Korea under the World Class University (WCU) program, for launch in November 2012 into a Sun-synchronous LEO to form TRIO-CINEMA. A fourth CINEMA is being developed for a 2013 launch into LEO. This LEO constellation of nanosatellites will provide unique measurements highly complementary to NASA's RBSP and THEMIS missions. Furthermore, CINEMA's development of miniature particle and magnetic field sensors, and cubesat-size spinning spacecraft may be important for future constellation space missions. Initial results from the first CINEMA will be presented if available.

  18. Comparison of Quantifiler(®) Trio and InnoQuant™ human DNA quantification kits for detection of DNA degradation in developed and aged fingerprints.

    PubMed

    Goecker, Zachary C; Swiontek, Stephen E; Lakhtakia, Akhlesh; Roy, Reena

    2016-06-01

    The development techniques employed to visualize fingerprints collected from crime scenes as well as post-development ageing may result in the degradation of the DNA present in low quantities in such evidence samples. Amplification of the DNA samples with short tandem repeat (STR) amplification kits may result in partial DNA profiles. A comparative study of two commercially available quantification kits, Quantifiler(®) Trio and InnoQuant™, was performed on latent fingerprint samples that were either (i) developed using one of three different techniques and then aged in ambient conditions or (ii) undeveloped and then aged in ambient conditions. The three fingerprint development techniques used were: cyanoacrylate fuming, dusting with black powder, and the columnar-thin-film (CTF) technique. In order to determine the differences between the expected quantities and actual quantities of DNA, manually degraded samples generated by controlled exposure of DNA standards to ultraviolet radiation were also analyzed. A total of 144 fingerprint and 42 manually degraded DNA samples were processed in this study. The results indicate that the InnoQuant™ kit is capable of producing higher degradation ratios compared to the Quantifiler(®) Trio kit. This was an expected result since the degradation ratio is a relative value specific for a kit based on the length and extent of amplification of the two amplicons that vary from one kit to the other. Additionally, samples with lower concentrations of DNA yielded non-linear relationships of degradation ratio with the duration of aging, whereas samples with higher concentrations of DNA yielded quasi-linear relationships. None of the three development techniques produced a noticeably different degradation pattern when compared to undeveloped fingerprints, and therefore do not impede downstream DNA analysis. PMID:27107968

  19. Effect of ursodeoxycholic acid treatment on the altered progesterone and bile acid homeostasis in the mother-placenta-foetus trio during cholestasis of pregnancy

    PubMed Central

    Estiú, Maria C; Monte, Maria J; Rivas, Laura; Moirón, Maria; Gomez-Rodriguez, Laura; Rodriguez-Bravo, Tomas; Marin, Jose JG; Macias, Rocio IR

    2015-01-01

    Aim Intrahepatic cholestasis of pregnancy (ICP) is characterized by pruritus and elevated bile acid concentrations in maternal serum. This is accompanied by an enhanced risk of intra-uterine and perinatal complications. High concentrations of sulphated progesterone metabolites (PMS) have been suggested to be involved in the multifactorial aetiopathogenesis of ICP. The aim of this study was to investigate further the mechanism accounting for the beneficial effect of oral administration of ursodeoxycholic acid (UDCA), which is the standard treatment, regarding bile acid and PMS homeostasis in the mother-placenta-foetus trio. Method Using HPLC-MS/MS bile acids and PMS were determined in maternal and foetal serum and placenta. The expression of ABC proteins in placenta was determined by real time quantitative PCR (RT-QPCR) and immunofluorescence. Results In ICP, markedly increased concentrations of bile acids (tauroconjugates > glycoconjugates >> unconjugated), progesterone and PMS in placenta and maternal serum were accompanied by enhanced concentrations in foetal serum of bile acids, but not of PMS. UDCA treatment reduced bile acid accumulation in the mother-placenta-foetus trio, but had no significant effect on progesterone and PMS concentrations. ABCG2 mRNA abundance was increased in placentas from ICP patients vs. controls and remained stable following UDCA treatment, despite an apparent further increase in ABCG2. Conclusion UDCA administration partially reduces ICP-induced bile acid accumulation in mothers and foetuses despite the lack of effect on concentrations of progesterone and PMS in maternal serum. Up-regulation of placental ABCG2 may play an important role in protecting the foetus from high concentrations of bile acids and PMS during ICP. PMID:25099365

  20. Performance Evaluation of Siemens ADVIA Centaur and Roche MODULAR Analytics E170 Total 25-OH Vitamin D Assays

    PubMed Central

    Chen, Yu; Kinney, Lois; Božović, Andrea; Smith, Hilary; Tarr, Heather; Diamandis, Eleftherios P.; LeBlanc, Adrien

    2014-01-01

    Objectives To evaluate the newly developed Roche MODULAR Analytics E170 Total Vitamin D and the Siemens ADVIA Centaur® Vitamin D Total assays. Materials and Methods Assays were evaluated using the Clinical and Laboratory Standards Institute protocols. Split patient samples were compared with LC-MS/MS and DiaSorin LIAISON assays (n=79 including 15 specimens with detectable endogenous 25-OH vitamin D2). Assay accuracy was also evaluated using the Vitamin D External Quality Assessment Scheme samples. Results The ADVIA Centaur and E170 assays demonstrated maximum total CVs of 14.1% and 5.9%, respectively. Both showed excellent linearity (R2 >0.99). The ADVIA Centaur assay demonstrated interference with bilirubin at 800 μmol/L, hemolysis at 1.25 g/L, and triglycerides at 2.8 mmol/L. Compared to LC-MS/MS, the ADVIA Centaur assay demonstrated a R2 value of 0.893, average bias of −8.8%; the E170 assay an R2 value of 0.872, average bias of 14.3% with underestimation of 25-OH vitamin D2. Compared to the LIAISON assay, the ADVIA Centaur assay demonstrated an R2 value of 0.781, average bias of −17.3%; the E170 assay an R2 value of 0.823, average bias of 11.4%. The ADVIA Centaur and E170 assays demonstrated a biases of <20% in 10/10 and 8/10 samples, respectively. Conclusions The ADVIA Centaur and E170 vitamin D assays demonstrated acceptable linearity, imprecision, and accuracy. The E170 assay demonstrated consistent underestimation of 25-OH vitamin D2 levels. Compared with LC-MS/MS, the ADVIA Centaur assay demonstrated a higher R2 value and a smaller average bias than the E170 assay. PMID:22705028

  1. Caveolin-3 T78M and T78K missense mutations lead to different phenotypes in vivo and in vitro.

    PubMed

    Traverso, Monica; Gazzerro, Elisabetta; Assereto, Stefania; Sotgia, Federica; Biancheri, Roberta; Stringara, Silvia; Giberti, Laura; Pedemonte, Marina; Wang, Xiabo; Scapolan, Sara; Pasquini, Elisabetta; Donati, Maria A; Zara, Federico; Lisanti, Michael P; Bruno, Claudio; Minetti, Carlo

    2008-03-01

    Caveolins are the principal protein components of caveolae, invaginations of the plasma membrane involved in cell signaling and trafficking. Caveolin-3 (Cav-3) is the muscle-specific isoform of the caveolin family and mutations in the CAV3 gene lead to a large group of neuromuscular disorders. In unrelated patients, we identified two distinct CAV3 mutations involving the same codon 78. Patient 1, affected by dilated cardiomyopathy and limb girdle muscular dystrophy (LGMD)-1C, shows an autosomal recessive mutation converting threonine to methionine (T78M). Patient 2, affected by isolated familiar hyperCKemia, shows an autosomal dominant mutation converting threonine to lysine (T78K). Cav-3 wild type (WT) and Cav-3 mutations were transiently transfected into Cos-7 cells. Cav-3 WT and Cav-3 T78M mutant localized at the plasma membrane, whereas Cav-3 T78K was retained in a perinuclear compartment. Cav-3 T78K expression was decreased by 87% when compared with Cav-3 WT, whereas Cav-3 T78M protein levels were unchanged. To evaluate whether Cav-3 T78K and Cav-3 T78M mutants behaved with a dominant negative pattern, Cos-7 cells were cotransfected with green fluorescent protein (GFP)-Cav-3 WT in combination with either mutant or WT Cav-3. When cotransfected with Cav-3 WT or Cav-3 T78M, GFP-Cav-3 WT was localized at the plasma membrane, as expected. However, when cotransfected with Cav-3 T78K, GFP-Cav-3 WT was retained in a perinuclear compartment, and its protein levels were reduced by 60%, suggesting a dominant negative action. Accordingly, Cav-3 protein levels in muscles from a biopsy of patient 2 (T78K mutation) were reduced by 80%. In conclusion, CAV3 T78M and T78K mutations lead to distinct disorders showing different clinical features and inheritance, and displaying distinct phenotypes in vitro. PMID:18253147

  2. Ursolic Acid Inhibits Adipogenesis in 3T3-L1 Adipocytes through LKB1/AMPK Pathway

    PubMed Central

    He, Yonghan; Li, Ying; Zhao, Tiantian; Wang, Yanwen; Sun, Changhao

    2013-01-01

    Background Ursolic acid (UA) is a triterpenoid compound with multiple biological functions. This compound has recently been reported to possess an anti-obesity effect; however, the mechanisms are less understood. Objective As adipogenesis plays a critical role in obesity, the present study was conducted to investigate the effect of UA on adipogenesis and mechanisms of action in 3T3-L1 preadipocytes. Methods and Results The 3T3-L1 preadipocytes were induced to differentiate in the presence or absence of UA for 6 days. The cells were determined for proliferation, differentiation, fat accumulation as well as the protein expressions of molecular targets that regulate or are involved in fatty acid synthesis and oxidation. The results demonstrated that ursolic acid at concentrations ranging from 2.5 µM to 10 µM dose-dependently attenuated adipogenesis, accompanied by reduced protein expression of CCAAT element binding protein β (C/EBPβ), peroxisome proliferator-activated receptor γ (PPARγ), CCAAT element binding protein α (C/EBPα) and sterol regulatory element binding protein 1c (SREBP-1c), respectively. Ursolic acid increased the phosphorylation of acetyl-CoA carboxylase (ACC) and protein expression of carnitine palmitoyltransferase 1 (CPT1), but decreased protein expression of fatty acid synthase (FAS) and fatty acid-binding protein 4 (FABP4). Ursolic acid increased the phosphorylation of AMP-activated protein kinase (AMPK) and protein expression of (silent mating type information regulation 2, homolog) 1 (Sirt1). Further studies demonstrated that the anti-adipogenic effect of UA was reversed by the AMPK siRNA, but not by the Sirt1 inhibitor nicotinamide. Liver kinase B1 (LKB1), the upstream kinase of AMPK, was upregulated by UA. When LKB1 was silenced with siRNA or the inhibitor radicicol, the effect of UA on AMPK activation was diminished. Conclusions Ursolic acid inhibited 3T3-L1 preadipocyte differentiation and adipogenesis through the LKB1/AMPK pathway

  3. Drug-induced Parkinsonism versus Idiopathic Parkinson Disease: Utility of Nigrosome 1 with 3-T Imaging.

    PubMed

    Sung, Young Hee; Noh, Young; Lee, Jongho; Kim, Eung Yeop

    2016-06-01

    Purpose To explore the utility of nigrosome 1 with 3-T magnetic resonance (MR) imaging to differentiate idiopathic Parkinson disease (IPD) from drug-induced parkinsonism (DIP). Materials and Methods The institutional review board approved this study, and participants gave informed consent. This study enrolled patients with DIP (n = 20) and IPD (n = 29) who underwent N-3-fluoropropyl-2-β-carbomethoxy-3-β-(4-iodophenyl)nortropane ((18)F-FP-CIT) positron emission tomography (PET) and healthy participants (n = 20). All participants underwent 0.5 × 0.5 × 1.0 mm(3) oblique axial three-dimensional multiecho-data image combination imaging to view the nigrosome 1 with 3-T imaging. Two reviewers independently assessed the nigrosome 1 without clinical information. DIP was diagnosed when no abnormality was seen at (18)F-FP-CIT PET. Diagnostic sensitivity, specificity, and accuracy of the nigrosome 1 imaging were evaluated between the IPD and DIP patients and between the IPD patients and healthy participants. Interrater agreement was assessed with Cohen κ. Results Both reviewers agreed in 63 of 69 participants (91.3%) for the presence of any abnormality on either side of the nigrosome 1 (κ = 0.825). Findings in all 29 IPD patients (100%) and three of 20 DIP patients (15%) were rated as abnormal and in 17 of 20 DIP patients (85%) they were interpreted as normal on the basis of imaging of the nitgrosome 1 (sensitivity, 100% (29 of 29); specificity, 85.0% (17 of 20); accuracy, 93.9% (46 of 49) between IPD and DIP patients). Findings in 3 of 20 healthy participants (15.0%) were interpreted as abnormal on the basis of imaging the nigrosome 1 while in the other 17 of 20 healthy participants (85.0%) they were rated as normal (sensitivity, 100% [29 of 29]; specificity, 85.0% [17 of 20]; accuracy, 93.9% [46 of 49] between IPD patients and healthy participants [κ = 0.831]). Conclusion The imaging of nigrosome 1 with 3-T imaging can differentiate DIP from IPD with high accuracy and

  4. Eight-channel transmit/receive body MRI coil at 3T.

    PubMed

    Vernickel, P; Röschmann, P; Findeklee, C; Lüdeke, K-M; Leussler, Ch; Overweg, J; Katscher, U; Grässlin, I; Schünemann, K

    2007-08-01

    Multichannel transmit magnetic resonance imaging (MR) systems have the potential to compensate for signal-intensity variations occurring at higher field strengths due to wave propagation effects in tissue. Methods such as RF shimming and local excitation in combination with parallel transmission can be applied to compensate for these effects. Moreover, parallel transmission can be applied to ease the excitation of arbitrarily shaped magnetization patterns. The implementation of these methods adds new requirements in terms of MRI hardware. This article describes the design of a decoupled eight-element transmit/receive body coil for 3T. The setup of the coil is explained, starting with standard single-channel resonators. Special focus is placed on the decoupling of the elements to obtain independent RF resonators. After a brief discussion of the underlying theory, the properties and limitations of the coil are outlined. Finally, the functionality and capabilities of the coil are demonstrated using RF measurements as well as MRI sequences. PMID:17654592

  5. Expression of an exogenous eukaryotic DNA methyltransferase gene induces transformation of NIH 3T3 cells.

    PubMed Central

    Wu, J; Issa, J P; Herman, J; Bassett, D E; Nelkin, B D; Baylin, S B

    1993-01-01

    Abnormal regional increases in DNA methylation, which have potential for causing gene inactivation and chromosomal instability, are consistently found in immortalized and tumorigenic cells. Increased DNA methyltransferase activity, which is also a characteristic of such cells, is a candidate to mediate these abnormal DNA methylation patterns. We now show that, in NIH 3T3 mouse fibroblasts, constitutive overexpression of an exogenous mouse DNA methyltransferase gene results in a marked increase in overall DNA methylation which is accompanied by tumorigenic transformation. These transformation changes can also be elicited by dexamethasone-inducible expression of an exogenous DNA methyltransferase gene. Our findings provide strong evidence that the increase in DNA methyltransferase activity associated with tumor progression could be a key step in carcinogenesis and provide a model system that can be used to further study this possibility. Images Fig. 1 Fig. 2 PMID:8415627

  6. Stimulation of protein phosphatase activity by insulin and growth factors in 3T3 cells

    SciTech Connect

    Chan, C.P.; McNall, S.J.; Krebs, E.G.; Fischer, E.H. )

    1988-09-01

    Incubation of Swiss mouse 3T3-D1 cells with physiological concentrations of insulin resulted in a rapid and transient activation of protein phosphatase activity as measure by using ({sup 32}P)phosphorylase {alpha} as substrate. Activation reached a maximum level (140% of control value) within 5 min of addition and returned to control levels within 20 min. The effect of insulin was dose-dependent with half-maximal activation occurring at {approx}5 nM insulin. This activity could be completely inhibited by addition of the heat-stable protein inhibitor 2, which suggests the presence of an activated type-1 phosphatase. Similar effects on phosphatase activity were seen when epidermal growth factor and platelet-derived growth factor were tested. These results suggest that some of the intracellular effects caused by insulin and growth factors are mediated through the activation of a protein phosphatase.

  7. Anthraquinones from Morinda officinalis roots enhance adipocyte differentiation in 3T3-L1 cells.

    PubMed

    Liu, Qing; Kim, Seon Beom; Ahn, Jong Hoon; Hwang, Bang Yeon; Kim, Sung Yeon; Lee, Mi Kyeong

    2012-01-01

    To search for anti-diabetic and insulin-sensitising natural products, the effect on adipocyte differentiation was investigated by assessing fat accumulation in 3T3-L1 preadipocytes using Oil Red O staining. Fractionation and separation of n-hexane and CHCl₃ fractions of Morinda officinalis (Rubiaceae) using several chromatographic methods led to the isolation of three anthraquinones, 1,2-dimethoxyanthraquinone (1), alizarin-2-methyl ether (2) and rubiadin-1-methyl ether (3). Among them, alizarin-2-methyl ether (2) showed the strongest enhancing activity, followed by rubiadin-1-methyl ether (3) and 1,2-dimethoxyanthraquinone (1). At a concentration of 100 µM, alizarin-2-methyl ether (2) enhanced adipocyte differentiation by up to 131% (compared to insulin-treated cells). Thus, these compounds could be beneficial in the treatment of diabetes. PMID:22008000

  8. Hepatitis C virus nonstructural protein NS3 transforms NIH 3T3 cells.

    PubMed Central

    Sakamuro, D; Furukawa, T; Takegami, T

    1995-01-01

    Clinical evidence suggests that hepatitis C virus (HCV) is etiologically involved in hepatic cancer and liver cirrhosis. To investigate whether the HCV nonstructural protein NS3 has oncogenic activity, NIH 3T3 cells were transfected with an expression vector containing cDNA for the 5'- or 3'-half sequence of the HCV genome segment encoding NS3. Only cells transfected with the 5'-half cDNA rapidly proliferated, lost contact inhibition, grew anchorage independently in soft agar, and formed tumors in nude mice. PCR analysis confirmed the presence of the 5'-half DNA in the transfectants. These results suggest that the 5' region of the HCV genome segment encoding NS3 is involved in cell transformation. PMID:7745741

  9. Prednisolone induces the Wnt signalling pathway in 3T3-L1 adipocytes

    PubMed Central

    Fleuren, Wilco W. M.; Linssen, Margot M. L.; Toonen, Erik J. M.; van der Zon, Gerard C. M.; Guigas, Bruno; de Vlieg, Jacob; Dokter, Wim H. A.; Ouwens, D. Margriet

    2013-01-01

    Synthetic glucocorticoids are potent anti-inflammatory drugs but show dose-dependent metabolic side effects such as the development of insulin resistance and obesity. The precise mechanisms involved in these glucocorticoid-induced side effects, and especially the participation of adipose tissue in this are not completely understood. We used a combination of transcriptomics, antibody arrays and bioinformatics approaches to characterize prednisolone-induced alterations in gene expression and adipokine secretion, which could underlie metabolic dysfunction in 3T3-L1 adipocytes. Several pathways, including cytokine signalling, Akt signalling, and Wnt signalling were found to be regulated at multiple levels, showing that these processes are targeted by prednisolone. These results suggest that mechanisms by which prednisolone induce insulin resistance include dysregulation of wnt signalling and immune response processes. These pathways may provide interesting targets for the development of improved glucocorticoids. PMID:23506355

  10. 3-T MRI detects inflammatory stenosis of the vertebral artery in giant cell arteritis.

    PubMed

    Geiger, J; Uhl, M; Peter, H H; Langer, M; Bley, T A

    2008-05-01

    Giant cell arteritis (GCA) is a granulomatous vasculitis. Early diagnosis is important for the initiation of corticosteroid treatment because the arteritis can result in blindness. In most of the cases, the superficial cranial arteries are affected. However, extracranial involvement of various arteries is known. Here, we report a case of histologically proven GCA with an inflammatory stenosis of the right vertebral artery. For complete evaluation of the extension of the disease, an optimized protocol of high-resolution magnetic resonance imaging at 3 T in combination with contrast-enhanced magnetic resonance angiography was performed. This non-invasive method facilitates the differentiation of inflamed and healthy segments of small cranial arteries, may help to find appropriate sites for biopsy, and allows the assessment of affected extracranial vessels. In this patient case, even the cause of vertebral stenosis--inflammatory versus arteriosclerotic--could be elucidated. PMID:18172573

  11. Reduction of 3T3 Fibroblast Adhesion on SS316L by Methyl-Terminated SAMs

    PubMed Central

    Raman, Aparna; Gawalt, Ellen S.

    2010-01-01

    Inhibiting the non-specific adhesion of cells and proteins to biomaterials such as stents, catheters and guide wires is an important interfacial issue that needs to be addressed in order to reduce surface-related implant complications. Medical grade stainless steel 316L was used as a model system to address this issue. To alter the interfacial property of the implant, self assembled monolayers of long chain phosphonic acids with −CH3, −COOH, −OH tail groups were formed on the native oxide surface of medical grade stainless steel 316L. The effect of varying the tail groups on 3T3 fibroblast adhesion was investigated. The methyl terminated phosphonic acid significantly prevented cell adhesion however presentation of hydrophilic tail groups at the interface did not significantly reduce cell adhesion when compared to the control stainless steel 316L. PMID:21461313

  12. Visual evoked potential (VEP) measured by simultaneous 64-channel EEG and 3T fMRI.

    PubMed

    Bonmassar, G; Anami, K; Ives, J; Belliveau, J W

    1999-06-23

    We present the first simultaneous measurements of evoked potentials (EPs) and fMRI hemodynamic responses to visual stimulation. Visual evoked potentials (VEPs) were recorded both inside and outside the static 3T magnetic field, and during fMRI examination. We designed, constructed, and tested a non-magnetic 64-channel EEG recording cap. By using a large number of EEG channels it is possible to design a spatial filter capable of removing the artifact noise present when recording EEG/EPs within a strong magnetic field. We show that the designed spatial filter is capable of recovering the ballistocardiogram-contaminated original EEG signal. Isopotential plots of the electrode array recordings at the peak of the VEP response (approximately 100ms) correspond well with simultaneous fMRI observed activated areas of primary and secondary visual cortices. PMID:10501528

  13. Kibizu concentrated liquid suppresses the accumulation of lipid droplets in 3T3-L1 cells.

    PubMed

    Inoue, Chisato; Kozaki, Tomomi; Morita, Yukiko; Shirouchi, Bungo; Fukami, Katsuya; Shimizu, Kuniyoshi; Sato, Masao; Katakura, Yoshinori

    2015-08-01

    Adipocyte size is closely related to the occurrence of diabetes, metabolic syndrome, and insulin resistance. Thus, researchers are searching for active substances that function to reduce adipocyte size. In the present study, we focused on sugar cane vinegar, Kibizu, and evaluated the function of Kibizu to reduce adipocyte size by using an in vitro model system, because people in Amami Oshima famous for longevity regularly consume Kibizu. Results showed that Kibizu treatment significantly reduced the size and number of lipid droplets in 3T3-L1 cells, relative to treatment with Kurozu, another traditional vinegar. Results of an extraction experiment suggest that the active components in Kibizu are lipophilic and hydrophobic. In addition, an in vivo experiment on rats treated with Kibizu showed that the active components were contained in large vein blood. Results of an additional in vivo experiment suggest that metabolites generated by Kibizu-treated rats are primarily contained or modified specifically in the large vein blood. PMID:25672941

  14. Measurement of brain glutamate using TE-averaged PRESS at 3T.

    PubMed

    Hurd, Ralph; Sailasuta, Napapon; Srinivasan, Radhika; Vigneron, Daniel B; Pelletier, Daniel; Nelson, Sarah J

    2004-03-01

    A method is introduced that provides improved in vivo spectroscopic measurements of glutamate (Glu), glutamine (Gln), choline (Cho), creatine (Cre), N-acetyl compounds (NAtot, NAA + NAAG), and the inositols (mI and sI). It was found that at 3T, TE averaging, the f1 = 0 slice of a 2D J-resolved spectrum, yielded unobstructed signals for Glu, Glu + Gln (Glx), mI, NA(tot), Cre, and Cho. The C4 protons of Glu at 2.35 ppm, and the C2 protons of Glx at 3.75 ppm were well resolved and yielded reliable measures of Glu/Gln stasis. Apparent T1/T2 values were obtained from the raw data, and metabolite tissue levels were determined relative to a readily available standard. A repeatibility error of <5%, and a coefficient of variation (CV) of <10% were observed for brain Glu levels in a study of six normal volunteers. PMID:15004781

  15. Regulation of p53 in NIH3T3 mouse fibroblasts following hyperosmotic stress.

    PubMed

    Lambert, Ian Henry; Enghoff, Maria Stine; Brandi, Marie-Luise; Hoffmann, Else Kay

    2015-06-01

    The aim of this project was to analyze the regulation of p53 expression in NIH3T3 fibroblasts under the influence of increasing hyperosmotic stress. Expression of p53 showed a biphasic response pattern in NIH3T3 cells under increasing osmotic stress (337 mOsm to 737 mOsm) with a maximum at 587 mOsm. Under isotonic conditions p53 expression increased after addition of the proteasome inhibitor MG132 indicating that cellular p53 levels in unperturbed cells is kept low by proteasomal degradation. However, under hypertonic conditions p53 synthesis as well as p53 degradation were significantly reduced and it is demonstrated that the increase in p53 expression observed when tonicity is increased from 337 to 587 mOsm reflects that degradation is more inhibited than synthesis, whereas the decrease in p53 expression at higher tonicities reflects that synthesis is more inhibited than degradation. The activity of the p53 regulating proteins p38 MAP kinase and the ubiquitin ligase MDM2 were studied as a function of increasing osmolarity. MDM2 protein expression was unchanged at all osmolarities, whereas MDM2 phosphorylation (Ser(166)) increased at osmolarities up to 537 mOsm and remained constant at higher osmolarities. Phosphorylation of p38 increased at osmolarities up to 687 mOsm which correlated with an increased phosphorylation of p53 (Ser(15)) and the decreased p53 degradation. Caspase-3 activity increased gradually with hypertonicity and at 737 mOsm both Caspase-3 activity and annexin V binding are high even though p53 expression and activity are low, indicating that initiation of apoptosis under severe hypertonic conditions is not strictly controlled by p53. PMID:26056062

  16. Carcinogenic potential of metal nanoparticles in BALB/3T3 cell transformation assay.

    PubMed

    Sighinolfi, G L; Artoni, E; Gatti, A M; Corsi, L

    2016-05-01

    Metal-based nanoparticles (NPs), are currently used in many application fields including consumer products, pharmaceuticals, and biomedical treatments. In spite to their wide applications, an in-depth study of their potential toxic effects is still lacking. The aim of the present research was to investigate the potential initiator or promoter-like activity of different metallic NPs such as gold, iron, cobalt, and cerium using the Balb/3T3 two-stage transformation assay. The results indicated that all the selected metallic NPs, except for cobalt, when used as initiators did not induce any transformation in Balb/3T3 cell line. Moreover, Au and Fe3 O4 NPs, when used in place of the tumor promoter treatment TPA, increased significantly the number of Foci/dish as compared to the MCA treatment alone. The number of Foci/dish was 2.6 for Au NPs and 2.13 for Fe3 O4 ones, similar to those obtained by the positive control treatment (MCA + TPA), whereas 1.27 for MCA treatment alone. On the contrary, CeO2 NPs did not show any difference in the number of Foci/dish, as compared to MCA alone, but it decreased the number of foci by 65% in comparison to the positive control (MCA + TPA). As expected, cobalt NPs showed an increased cytotoxicity and only a few surviving cells were found at the time of analysis showing a number of Foci/dish of 0.13. For the first time, our data clearly showed that Au and Fe3 O4 NPs act as promoters in the two stage transformational assay, suggesting the importance to fully investigate the NPs carcinogenic potential with different models. PMID:25358123

  17. Superior Cervical Sympathetic Ganglion: Normal Imaging Appearance on 3T-MRI

    PubMed Central

    Lee, Joo Yeon; Song, Joon Seon; Song, Min Jeong; Hwang, Seung-Jun; Yoon, Ra Gyoung; Jang, Seung Won; Park, Ji Eun; Heo, Young Jin; Choi, Young Jun; Baek, Jung Hwan

    2016-01-01

    Objective To identify superior cervical sympathetic ganglion (SCSG) and describe their characteristic MR appearance using 3T-MRI. Materials and Methods In this prospective study, we recruited 53 consecutive patients without history of head and neck irradiation. Using anatomic location based on literature review, both sides of the neck were evaluated to identify SCSGs in consensus. SCSGs were divided into definite (medial to internal carotid artery [ICA] and lateral to longus capitis muscle [LCM]) and probable SCSGs based on relative location to ICA and LCM. Two readers evaluated signal characteristics including intraganglionic hypointensity of all SCSGs and relative location of probable SCSGs. Interrater and intrarater agreements were quantified using unweighted kappa. Results Ninety-one neck sites in 53 patients were evaluated after exclusion of 15 neck sites with pathology. Definite SCSGs were identified at 66 (73%) sites, and probable SCSGs were found in 25 (27%). Probable SCSGs were located anterior to LCM in 16 (18%), lateral to ICA in 6 (7%), and posterior to ICA in 3 (3%). Intraganglionic hypointensity was identified in 82 (90%) on contrast-enhanced fat-suppressed T1-weighted images. There was no statistical difference in the relative location between definite and probable SCSGs of the right and left sides with intragnalionic hypointensity on difference pulse sequences. Interrater and intrarater agreements on the location and intraganglionic hypointensity were excellent (κ-value, 0.749–1.000). Conclusion 3T-MRI identified definite SCSGs at 73% of neck sites and varied location of the remaining SCSGs. Intraganglionic hypointensity was a characteristic feature of SCSGs. PMID:27587954

  18. Regulation of p53 in NIH3T3 mouse fibroblasts following hyperosmotic stress

    PubMed Central

    Lambert, Ian Henry; Enghoff, Maria Stine; Brandi, Marie-Luise; Hoffmann, Else Kay

    2015-01-01

    The aim of this project was to analyze the regulation of p53 expression in NIH3T3 fibroblasts under the influence of increasing hyperosmotic stress. Expression of p53 showed a biphasic response pattern in NIH3T3 cells under increasing osmotic stress (337 mOsm to 737 mOsm) with a maximum at 587 mOsm. Under isotonic conditions p53 expression increased after addition of the proteasome inhibitor MG132 indicating that cellular p53 levels in unperturbed cells is kept low by proteasomal degradation. However, under hypertonic conditions p53 synthesis as well as p53 degradation were significantly reduced and it is demonstrated that the increase in p53 expression observed when tonicity is increased from 337 to 587 mOsm reflects that degradation is more inhibited than synthesis, whereas the decrease in p53 expression at higher tonicities reflects that synthesis is more inhibited than degradation. The activity of the p53 regulating proteins p38 MAP kinase and the ubiquitin ligase MDM2 were studied as a function of increasing osmolarity. MDM2 protein expression was unchanged at all osmolarities, whereas MDM2 phosphorylation (Ser166) increased at osmolarities up to 537 mOsm and remained constant at higher osmolarities. Phosphorylation of p38 increased at osmolarities up to 687 mOsm which correlated with an increased phosphorylation of p53 (Ser15) and the decreased p53 degradation. Caspase-3 activity increased gradually with hypertonicity and at 737 mOsm both Caspase-3 activity and annexin V binding are high even though p53 expression and activity are low, indicating that initiation of apoptosis under severe hypertonic conditions is not strictly controlled by p53. PMID:26056062

  19. In-bore setup and software for 3T MRI-guided transperineal prostate biopsy

    NASA Astrophysics Data System (ADS)

    Tokuda, Junichi; Tuncali, Kemal; Iordachita, Iulian; Song, Sang-Eun; Fedorov, Andriy; Oguro, Sota; Lasso, Andras; Fennessy, Fiona M.; Tempany, Clare M.; Hata, Nobuhiko

    2012-09-01

    MRI-guided prostate biopsy in conventional closed-bore scanners requires transferring the patient outside the bore during needle insertion due to the constrained in-bore space, causing a safety hazard and limiting image feedback. To address this issue, we present our custom-made in-bore setup and software to support MRI-guided transperineal prostate biopsy in a wide-bore 3 T MRI scanner. The setup consists of a specially designed tabletop and a needle-guiding template with a Z-frame that gives a physician access to the perineum of the patient at the imaging position and allows the physician to perform MRI-guided transperineal biopsy without moving the patient out of the scanner. The software and Z-frame allow registration of the template, target planning and biopsy guidance. Initially, we performed phantom experiments to assess the accuracy of template registration and needle placement in a controlled environment. Subsequently, we embarked on our clinical trial (N = 10). The phantom experiments showed that the translational errors of the template registration along the right-left (RP) and anterior-posterior (AP) axes were 1.1 ± 0.8 and 1.4 ± 1.1 mm, respectively, while the rotational errors around the RL, AP and superior-inferior axes were (0.8 ± 1.0)°, (1.7 ± 1.6)° and (0.0 ± 0.0)°, respectively. The 2D root-mean-square (RMS) needle-placement error was 3 mm. The clinical biopsy procedures were safely carried out in all ten clinical cases with a needle-placement error of 5.4 mm (2D RMS). In conclusion, transperineal prostate biopsy in a wide-bore 3T scanner is feasible using our custom-made tabletop setup and software, which supports manual needle placement without moving the patient out of the magnet.

  20. Melatonin Suppresses Autophagy Induced by Clinostat in Preosteoblast MC3T3-E1 Cells

    PubMed Central

    Yoo, Yeong-Min; Han, Tae-Young; Kim, Han Sung

    2016-01-01

    Microgravity exposure can cause cardiovascular and immune disorders, muscle atrophy, osteoporosis, and loss of blood and plasma volume. A clinostat device is an effective ground-based tool for simulating microgravity. This study investigated how melatonin suppresses autophagy caused by simulated microgravity in preosteoblast MC3T3-E1 cells. In preosteoblast MC3T3-E1 cells, clinostat rotation induced a significant time-dependent increase in the levels of the autophagosomal marker microtubule-associated protein light chain (LC3), suggesting that autophagy is induced by clinostat rotation in these cells. Melatonin treatment (100, 200 nM) significantly attenuated the clinostat-induced increases in LC3 II protein, and immunofluorescence staining revealed decreased levels of both LC3 and lysosomal-associated membrane protein 2 (Lamp2), indicating a decrease in autophagosomes. The levels of phosphorylation of mammalian target of rapamycin (p-mTOR) (Ser2448), phosphorylation of extracellular signal-regulated kinase (p-ERK), and phosphorylation of serine-threonine protein kinase (p-Akt) (Ser473) were significantly reduced by clinostat rotation. However, their expression levels were significantly recovered by melatonin treatment. Also, expression of the Bcl-2, truncated Bid, Cu/Zn- superoxide dismutase (SOD), and Mn-SOD proteins were significantly increased by melatonin treatment, whereas levels of Bax and catalase were decreased. The endoplasmic reticulum (ER) stress marker GRP78/BiP, IRE1α, and p-PERK proteins were significantly reduced by melatonin treatment. Treatment with the competitive melatonin receptor antagonist luzindole blocked melatonin-induced decreases in LC3 II levels. These results demonstrate that melatonin suppresses clinostat-induced autophagy through increasing the phosphorylation of the ERK/Akt/mTOR proteins. Consequently, melatonin appears to be a potential therapeutic agent for regulating microgravity-related bone loss or osteoporosis. PMID:27070587

  1. Mobile phone base station radiation does not affect neoplastic transformation in BALB/3T3 cells.

    PubMed

    Hirose, H; Suhara, T; Kaji, N; Sakuma, N; Sekijima, M; Nojima, T; Miyakoshi, J

    2008-01-01

    A large-scale in vitro study focusing on low-level radiofrequency (RF) fields from mobile radio base stations employing the International Mobile Telecommunication 2000 (IMT-2000) cellular system was conducted to test the hypothesis that modulated RF fields affect malignant transformation or other cellular stress responses. Our group previously reported that DNA strand breaks were not induced in human cells exposed to 2.1425 GHz Wideband Code Division Multiple Access (W-CDMA) radiation up to 800 mW/kg from mobile radio base stations employing the IMT-2000 cellular system. In the current study, BALB/3T3 cells were continuously exposed to 2.1425 GHz W-CDMA RF fields at specific absorption rates (SARs) of 80 and 800 mW/kg for 6 weeks and malignant cell transformation was assessed. In addition, 3-methylcholanthrene (MCA)-treated cells were exposed to RF fields in a similar fashion, to assess for effects on tumor promotion. Finally, the effect of RF fields on tumor co-promotion was assessed in BALB/3T3 cells initiated with MCA and co-exposed to 12-O-tetradecanoylphorbol-13-acetate (TPA). At the end of the incubation period, transformation dishes were fixed, stained with Giemsa, and scored for morphologically transformed foci. No significant differences in transformation frequency were observed between the test groups exposed to RF signals and the sham-exposed negative controls in the non-, MCA-, or MCA plus TPA-treated cells. Our studies found no evidence to support the hypothesis that RF fields may affect malignant transformation. Our results suggest that exposure to low-level RF radiation of up to 800 mW/kg does not induce cell transformation, which causes tumor formation. PMID:17694516

  2. Oleanolic acid reduces markers of differentiation in 3T3-L1 adipocytes.

    PubMed

    Sung, Hye-Young; Kang, Sang-Wook; Kim, Jung-Lye; Li, Jing; Lee, Eun-Sook; Gong, Ju-Hyun; Han, Seoung Jun; Kang, Young-Hee

    2010-12-01

    Oleanolic acid is a triterpenoid compound that is widely present in vegetables, medicinal herbs, and other plants and has potent antioxidant and antiinflammatory properties. However, the potential of oleanolic acid to offset obesity is not clear. This study tested the hypothesis that oleanolic acid suppresses the differentiation of 3T3-L1 adipocytes by downregulating cellular induction of peroxisome proliferators-activated receptor γ (PPARγ) and cytidine-cytidine-adenosine-adenosine-thymidine (CCAAT) enhancer binding protein α (C/EBPα). The 3T3-L1 adipocytes were cultured and differentiated in Dulbecco modified Eagle medium containing 10% fetal bovine serum for 6 to 8 days in the absence and presence of 1 to 25 μmol/L oleanolic acid according to differentiating protocols. Nontoxic oleanolic acid, at 25 μmol/L or less, dose-dependently attenuated lipid accumulation in differentiated adipocytes as evidenced by Oil Red O staining. Western blot analysis showed that the induction of PPARγ and C/EBPα was markedly attenuated in differentiated and oleanolic acid-treated adipocytes at their transcriptional messenger RNA levels. Furthermore, this study examined whether oleanolic acid dampened the induction of visfatin, a proinflammatory and visceral fat-specific adipokine expressed in adipocytes. Visfatin expression was inhibited in differentiated adipocytes exposed to a PPARγ inhibitor GW9662. In addition, the visfatin production was significantly repressed in 25 μmol/L oleanolic acid-treated adipocytes, possibly through blocking PPARγ activation. These results demonstrate that oleanolic acid may be a promising agent to disturb adipocyte differentiation and suppress obesity-associated inflammation. PMID:21147366

  3. Mechanisms and prevention of thermal injury from gamma radiosurgery headframes during 3T MR imaging.

    PubMed

    Bennett, Marcus C; Wiant, David B; Gersh, Jacob A; Dolesh, Wendy; Ding, X; Best, Ryan C M; Bourland, J D

    2012-01-01

    Magnetic resonance imaging (MRI) is regularly used for stereotactic imaging of Gamma Knife (GK) radiosurgery patients for GK treatment planning. MRI-induced thermal injuries have occurred and been reported for GK patients with attached metallic headframes. Depending on the specific MR imaging and headframe conditions, a skin injury from MRI-induced heating can potentially occur where the four headframe screws contact the skin surface of the patient's head. Higher MR field strength has a greater heating potential. Two primary heating mechanisms, electromagnetic induction and the antenna effect, are possible. In this study, MRI-induced heating from a 3T clinical MRI scanner was investigated for stereotactic headframes used in gamma radiosurgery and neurosurgery. Using melons as head phantoms, optical thermometers were used to characterize the temperature profile at various points of the melon headframe composite as a function of two 3T MR pulse sequence protocols. Different combinations of GK radiosurgery headframe post and screw designs were tested to determine best and worst combinations for MRI-induced heating. Temperature increases were measured for all pulse sequences tested, indicating that the potential exists for MRI-induced skin heating and burns at the headframe attachment site. This heating originates with electromagnetic induction caused by the RF fields inducing current in a loop formed by the headframe, mounting screws, and the region of the patient's head located between any of the two screws. This induced current is then resistively dissipated, with the regions of highest resistance, located at the headframe screw-patient head interface, experiencing the most heating. Significant heating can be prevented by replacing the metallic threads holding the screw with electrically insulated nuts, which is the heating prevention and patient safety recommendation of the GK manufacturer. Our results confirm that the manufacturer's recommendation to use

  4. In vitro BALB/3T3 cell transformation assay of nonoxynol-9 and 1,4-dioxane

    SciTech Connect

    Sheu, C.W.; Moreland, F.M.; Lee, J.K.; Dunkel, V.C.

    1988-01-01

    The spermicidal surfactant nonoxynol-9 (Igepal CO-630, GAF Corp.) and a potential impurity, 1,4-dioxane, were tested in the in vitro cell transformation assay using BALB/3T3 cells. Two treatment periods, 48 hr and 13 days, were used. Nonoxynol-9, tested at levels up to 10 /sup +/g/ml, did not induce transformation, whereas dioxane was very active in the induction type II foci in the cultured BALB/3T3 cells.

  5. Suppressed intrinsic catalytic activity of GLUT1 glucose transporters in insulin-sensitive 3T3-L1 adipocytes

    SciTech Connect

    Harrison, S.A.; Buxton, J.M.; Czech, M.P. )

    1991-09-01

    Previous studies indicated that the erythroid-type (GLUT1) glucose transporter isoform contributes to basal but not insulin-stimulated hexose transport in mouse 3T3-L1 adipocytes. In the present studies it was found that basal hexose uptake in 3T3-L1 adipocytes was about 50% lower than that in 3T3-L1 or CHO-K1 fibroblasts. Intrinsic catalytic activities of GLUT1 transporters in CHO-K1 and 3T3-L1 cells were compared by normalizing these hexose transport rates to GLUT1 content on the cell surface, as measured by two independent methods. Cell surface GLUT1 levels in 3T3-L1 fibroblasts and adipocytes were about 10- and 25-fold higher, respectively, than in CHO-K1 fibroblasts, as assessed with an anti-GLUT1 exofacial domain antiserum, delta. The large excess of cell surface GLUT1 transporters in 3T3-L1 adipocytes relative to CHO-K1 fibroblasts was confirmed by GLUT1 protein immunoblot analysis and by photoaffinity labeling (with 3-({sup 125}I)iodo-4-azidophenethylamido-7-O-succinyldeacetylforskolin) of glucose transporters in isolated plasma membranes. Thus, GLUT1 intrinsic activity is markedly reduced in 3T3-L1 fibroblasts compared with the CHO-K1 fibroblasts, and further reduction occurs upon differentiation to adipocytes. The authors conclude that a mechanism that markedly suppresses basal hexose transport catalyzed by GLUT1 is a major contributor to the dramatic insulin sensitivity of glucose uptake in 3T3-L1 adipocytes.

  6. Goldvalve detachable balloon: "in vitro" assessment of safety and imaging artifacts in a 3-T MR system.

    PubMed

    Dabus, Guilherme; Gerstle, Ronald J; Goodman, James; Cross, DeWitte T; Moran, Christopher J; Derdeyn, Colin P

    2006-06-01

    The Goldvalve balloon is the only currently available detachable balloon. We undertook a study to determine safety and imaging artifacts in a MR environment. We found no evidence for heating or deflection of the balloon in a comprehensive series of in vitro experiments at 3 T. MR imaging at field strengths up to 3 T of patients with implanted Goldvalve balloons is safe. Imaging artifacts are minimal. PMID:16758155

  7. SU-C-201-07: Validation of a GATE Gamma Camera Model for the Siemens Symbia

    SciTech Connect

    Mikell, J; Siman, W; Kappadath, S; Mourtada, F

    2015-06-15

    Purpose: To develop a simulation model of a clinical gamma camera/SPECT system and to validate the model using experimental and published measurements from the clinical system. Methods: Geant4 Application for Tomographic Emission (GATE) was used to create a model of the Siemens Symbia gamma camera. A modular model was implemented that allows specifying combinations of crystal thickness (3/8”, 5/8”) and collimator (LEHR, MELP, HE). Shielding, energy resolution, intrinsic resolution, crystal thickness, and collimator properties were set based on manufacturer specifications. Validation of the model was performed by simulating NEMA 2007 gamma camera tests including spatial resolution and sensitivity for Tc99; these were compared with experimental and published data for the scanner. The simulated energy spectra of a Tc99 line source in acrylic blocks was visually compared with the corresponding experimental acquisition. For a 4 cm diameter sphere filled with Tc99, the attenuation maps were generated from simulation data, and the photopeak and scatter window were extracted from GATE output using ROOT to create DICOM files to use in the clinical reconstruction. Results: Simulated spatial resolutions for LEHR 3/8” crystal at 0, 10 cm, 10 cm (with scatter), and 30 cm were 4, 6.7, 7.9, and 14.5 mm FWHM; these were 9% less than published data. For 5/8” crystal the spatial resolutions were 4.5, 7.0, 8.5, and 14.7 mm FWHM; these were 4% to 10% less than published data. Simulated sensitivity was within 3.5% of published data for both LEHR 3/8” and 5/8”. The simulated energy spectra matched the photopeak and scatter window well, but did overestimate the counts below 90 keV. The simulated attenuation map and projection data were successfully reconstructed with the clinical software, and the passed visual inspection. Conclusions: Validation of a specific clinical scanner allows future studies of quantification accuracy for both planar and SPECT imaging. Research

  8. Insulin regulation of protein biosynthesis in differentiated 3T3 adipocytes. Regulation of glyceraldehyde-3-phosphate dehydrogenase

    SciTech Connect

    Alexander, M.; Curtis, G.; Avruch, J.; Goodman, H.M.

    1985-10-05

    The effect of insulin on protein biosynthesis was examined in differentiated 3T3-L1 and 3T3-F442A adipocytes. Insulin altered the relative rate of synthesis of specific proteins independent of its ability to hasten conversion of the fibroblast (preadipocyte) phenotype to the adipocyte phenotype. Although more than one pattern of response to insulin was observed, the authors focused on the induction of a Mr 33,000 protein which was identified as the glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Exposure of 3T3 adipocytes to insulin throughout differentiation specifically increased GAPDH activity and protein content by 2- to 3-fold as compared to 3T3 adipocytes differentiated in the absence of insulin. These changes in enzyme activity and content could be accounted for by a 4-fold increase in the relative rate of synthesis of GAPDH and a 9-fold increase in hybridizable mRNA levels. Within 2 h of insulin addition to 3T3 adipocytes differentiated in the absence of hormone, hybridizable GAPDH mRNA levels increased 3-fold, and within 24 h GAPDH mRNA levels increased 8-fold, and (TVS) methionine incorporation into GAPDH protein increased 5-fold. These studies demonstrate that insulin, as the sole hormonal perturbant, can increase the synthesis of certain 3T3 adipocyte proteins by altering the cellular content of a specific mRNA.

  9. Anti-obesity effect of Blumea balsamifera extract in 3T3-L1 preadipocytes and adipocytes.

    PubMed

    Kubota, Hiroaki; Kojima-Yuasa, Akiko; Morii, Risako; Huang, Xuedan; Norikura, Toshio; Rho, Sook-Nyung; Matsui-Yuasa, Isao

    2009-01-01

    Obesity, the leading metabolic disease in the world, is a serious health problem in industrialized countries. We investigated the anti-obesity effect of Blumea balsamifera extract on adipocyte differentiation of 3T3-L1 preadipocytes and anti-obesity effect of 3T3-L1 adipocytes. We found that treatment with an extract of Blumea balsamifera suppressed lipid accumulation and glycerol-3-phosphate dehydrogenase (GPDH) activity without affecting cell viability in 3T3-L1 preadipocytes and adipocytes. Furthermore, Blumea balsamifera extract brought significant attenuation of expressions of key adipogenic transcription factors, including peroxisome proliferator-activated receptor (PPAR)gamma, CCAAT element binding protein (C/EBPs) and leptin, however, induced up-regulation of adiponectin at the protein level in 3T3-L1 preadipocytes and adipocytes. These results suggest that Blumea balsamifera extract may block adipogenesis, at least in part, by decreasing key adipogenic transcription factors in 3T3-L1 preadipocytes and may have antiatherogenic, anti-inflammatory, and antidiabetic effects through up-regulation of adiponectin in 3T3-L1 adipocytes. PMID:19885945

  10. Resistance to oncogenic transformation in revertant R1 of human ras-transformed NIH 3T3 cells

    SciTech Connect

    Kuzumaki, N.; Ogiso, Y.; Oda, A.; Fujita, H.; Suzuki, H.; Sato, C.; Mullauer, L.

    1989-05-01

    A flat revertant, R1, was isolated from human activated c-Ha-ras-1 (hu-ac-Ha-ras) gene-transformed NIH 3T3 cells (EJ-NIH 3T3) treated with mutagens. R1 contained unchanged transfected hu-ac-Ha-ras DNA and expressed high levels of hu-ac-Ha-ras-specific mRNA and p21 protein. Transfection experiments revealed that NIH 3T3 cells could be transformed by DNA from R1 cells but R1 cells could not be retransformed by Kirsten sarcoma virus, DNA from EJ-NIH 3T3 cells, hu-ac-Ha-ras, v-src, v-mos, simian virus 40 large T antigen, or polyomavirus middle T antigen. Somatic cell hybridization studies showed that R1 was not retransformed by fusion with NIH 3T3 cells and suppressed anchorage independence of EJ-NIH 3T3 and hu-ac-Ha-ras gene-transformed rat W31 cells in soft agar. These results suggest that the reversion and resistance to several oncogenes in R1 is due n not to cellular defects in the production of the transformed phenotype but rather to enhancement of cellular mechanisms that suppress oncogenic transformation.

  11. Coculture with BJ fibroblast cells inhibits the adipogenesis and lipogenesis in 3T3-L1 cells

    SciTech Connect

    Jeong, Hyun Jeong; Park, Sahng Wook; Kim, Hojeong; Park, Sang-Kyu; Yoon, Dojun

    2010-02-19

    Mouse or human fibroblasts are commonly used as feeder cells to prevent differentiation in stem or primary cell culture. In the present study, we addressed whether fibroblasts can affect the differentiation of adipocytes. We found that the differentiation of 3T3-L1 preadipocytes was strongly suppressed when the cells were cocultured with human fibroblast (BJ) cells. BrdU incorporation analysis indicated that mitotic clonal expansion, an early event required for 3T3-L1 cell adipogenesis, was not affected by BJ cells. The 3T3-L1 cell expression levels of peroxisome proliferator-activated receptor {gamma}2, CCAAT/enhancer-binding protein alpha (C/EBP{alpha}), sterol regulatory element binding protein-1c, and Krueppel-like factor 15, but not those of C/EBP{beta} or C/EBP{delta}, were decreased by coculture with BJ cells. When mature 3T3-L1 adipocytes were cocultured with BJ cells, their lipid contents were significantly reduced, with decreased fatty acid synthase expression and increased phosphorylated form of acetyl-CoA carboxylase 1. Our data indicate that coculture with BJ fibroblast cells inhibits the adipogenesis of 3T3-L1 preadipocytes and decreases the lipogenesis of mature 3T3-L1 adipocytes.

  12. ATF3 inhibits adipocyte differentiation of 3T3-L1 cells

    SciTech Connect

    Jang, Min Kyung; Kim, Cho Hee; Seong, Je Kyung; Jung, Myeong Ho

    2012-04-27

    Highlights: Black-Right-Pointing-Pointer Overexpression of ATF3 inhibits adipocyte differentiation in 3T3-L1 cells. Black-Right-Pointing-Pointer Overexpression of ATF3 represses C/EBP{alpha} expression. Black-Right-Pointing-Pointer ATF3 directly binds to mouse C/EBP{alpha} promoter spanning from -1928 to -1907. Black-Right-Pointing-Pointer ATF3 may play a role in hypoxia-mediated inhibition of adipocyte differentiation. -- Abstract: ATF3 is a stress-adaptive gene that regulates proliferation or apoptosis under stress conditions. However, the role of ATF3 is unknown in adipocyte cells. Therefore, in this study, we investigated the functional role of ATF3 in adipocytes. Both lentivirus-mediated overexpression of ATF3 and stably-overexpressed ATF3 inhibited adipocyte differentiation in 3T3-L1 cells, as revealed by decreased lipid staining with oil red staining and reduction in adipogenic genes. Thapsigargin treatment and overexpression of ATF3 decreased C/EBP{alpha} transcript and repressed the activity of the 3.6-kb mouse C/EBP{alpha} promoter, demonstrating that ATF3 downregulates C/EBP{alpha} expression. Transfection studies using mutant constructs containing 5 Prime -deletions in the C/EBP{alpha} promoter revealed that a putative ATF/CRE element, GGATGTCA, is located between -1921 and -1914. Electrophoretic mobility shift assay and chromatin immunoprecipitation assay demonstrated that ATF3 directly binds to mouse C/EBP{alpha} promoter spanning from -1928 to -1907. Both chemical hypoxia-mimetics or physical hypoxia led to reduce the C/EBP{alpha} mRNA and repress the promoter activity of the C/EBP{alpha} gene, whereas increase ATF3 mRNA, suggesting that ATF3 may contribute to the inhibition of adipocyte differentiation in hypoxia through downregulation of C/EBP{alpha} expression. Collectively, these results demonstrate that ATF3 represses the C/EBP{alpha} gene, resulting in inhibition of adipocyte differentiation, and thus plays a role in hypoxia-mediated inhibition

  13. Nitric Oxide-Induced Autophagy in MC3T3-E1 Cells is Associated with Cytoprotection via AMPK Activation

    PubMed Central

    Yang, Jung Yoon; Park, Min Young; Park, Sam Young; Yoo, Hong Il; Kim, Min Seok; Kim, Jae Hyung

    2015-01-01

    Nitric oxide (NO) is important in the regulation of bone remodeling, whereas high concentration of NO promotes cell death of osteoblast. However, it is not clear yet whether NO-induced autophagy is implicated in cell death or survival of osteoblast. The present study is aimed to examine the role of NO-induced autophagy in the MC3T3-E1 cells and their underlying molecular mechanism. The effect of sodium nitroprusside (SNP), an NO donor, on the cytotoxicity of the MC3T3-E1 cells was determined by MTT assay and expression of apoptosis or autophagy associated molecules was evaluated by western blot analysis. The morphological observation of autophagy and apoptosis by acridine orange stain and TUNEL assay were performed, respectively. Treatment of SNP decreased the cell viability of the MC3T3-E1 cells in dose- and time-dependent manner. SNP increased expression levels of p62, ATG7, Beclin-1 and LC3-II, as typical autophagic markers and augmented acidic autophagolysosomal vacuoles, detected by acridine orange staining. However, pretreatment with 3-methyladenine (3MA), the specific inhibitor for autophagy, decreased cell viability, whereas increased the cleavage of PARP and caspase-3 in the SNP-treated MC3T3-E1 cells. AMP-activated protein kinase (AMPK), a major autophagy regulatory kinase, was activated in SNP-treated MC3T3-E1 cells. In addition, pretreatment with compound C, an inhibitor of AMPK, decreased cell viability, whereas increased the number of apoptotic cells, cleaved PARP and caspase-3 levels compared to those of SNP-treated MC3T3-E1 cells. Taken together, it is speculated that NO-induced autophagy functions as a survival mechanism via AMPK activation against apoptosis in the MC3T3-E1 cells. PMID:26557017

  14. Evidence that downregulation of hexose transport limits intracellular glucose in 3T3-L1 fibroblasts

    SciTech Connect

    Whitesell, R.R.; Regen, D.M.; Pelletier, D.; Abumrad, N.A. )

    1990-10-01

    Measurements of initial glucose entry rate and intracellular glucose concentration in cultured cells are difficult because of rapid transport relative to intracellular volume and a substantial extracellular space from which glucose cannot be completely removed by quick exchanges of medium. In 3T3-L1 cells, we obtained good estimates of initial entry of ({sup 14}C)methylglucose and D-({sup 14}C)glucose with (1) L-({sup 3}H)glucose as an extracellular marker together with the ({sup 14}C)glucose or ({sup 14}C)methylglucose in the substrate mixture, (2) sampling times as short as 2 s, (3) ice-cold phloretin-containing medium to stop uptake and rinse away the extracellular label, and (4) nonlinear regression of time courses. Methylglucose equilibrated in two phases--the first with a half-time of 1.7 s and the second with a half-time of 23 s; it eventually equilibrated in an intracellular space of 8 microliters/mg protein. Entry of glucose remained almost linear for 10 s, making its transport kinetics easier to study (Km = 5.7 mM, Vmax = 590 nmol.s-1.ml-1 cell water). Steady-state intracellular glucose concentration was 75-90% of extracellular glucose concentration. Cells grown in a high-glucose medium (24 mM) exhibited a 67% reduction of glucose-transport activity and a 50% reduction of steady-state ratio of intracellular glucose to extracellular glucose.

  15. Increased Visibility of Deep Medullary Veins in Leukoaraiosis: A 3-T MRI Study

    PubMed Central

    Yan, Shenqiang; Wan, Jinping; Zhang, Xuting; Tong, Lusha; Zhao, Song; Sun, Jianzhong; Lin, Yuehan; Shen, Chunhong; Lou, Min

    2013-01-01

    Cerebral venous collagenosis has been implicated in leading to white matter hyperintensities (WMHs) via venous ischemia. We sought to determine whether cerebral venous dilation or ischemia correlate with the severity of WMHs by quantitative in vivo imaging techniques. This was an investigator-initiated prospective single-center study. We reviewed clinical, laboratory data from 158 consecutive WMHs patients and 50 controls, and measured the number of voxels of deep medullary veins (DMVs) on susceptibility-weighted image and assessed the WMH volume (as a marker of the severity of WMHs) on a 3-T magnetic resonance system. We then performed the logistic-regression analysis and partial Pearson’s correlation analysis to examine the association between the venous voxel count and WMH volume. The number of voxels of DMVs was significantly higher in WMHs than in controls. Increased number of voxels of DMVs was independently associated with both WMH volume of the whole brain and coregistered regional WMH volume after adjusting for age and number of lacunes. Our study indicates that cerebral deep venous insufficiency or ischemia play a role in the pathogenesis of WMHs, which may provide prognostic information on patients with WMHs and may have implications for therapeutic interventions. PMID:25071553

  16. Shikonin inhibits fat accumulation in 3T3-L1 adipocytes.

    PubMed

    Lee, Haeyong; Kang, Ryunhwa; Yoon, Yoosik

    2010-03-01

    Shikonin, 5,6-dihydroxyflavone-7-glucuronic acid, is the main ingredient of Lithospermum erythrorhizon Sieb. et Zucc, and was reported to have various biological activities including antiinflammatory, anticancer, antimicrobial and others. This study aimed to elucidate, for the first time, the antiobesity activity of shikonin and its mechanism of action. Shikonin was found to inhibit fat droplet formation and triglyceride accumulation in 3T3-L1 adipocytes. The half inhibitory concentration, IC(50), for the inhibition of triglyceride accumulation was found to be 1.1 microM. The expression of genes involved in lipid metabolism, such as FABP4 and LPL, were significantly inhibited following shikonin treatment. Shikonin also inhibited the ability of PPAR gamma and C/EBP alpha, the major transcription factors of adipogenesis, to bind to their target DNA sequences. The expressions of mRNA and protein of PPAR gamma and C/EBPa were significantly down-regulated following shikonin treatment. Among the upstream regulators of adipogenesis, only SREBP1C was found to be down-regulated by shikonin. The results of this study suggest that shikonin down-regulates the expression of SREBP1C and subsequently the expression of PPAR gamma and C/EBP alpha. Together, these changes result in the down-regulation of lipid metabolizing enzymes and reduced fat accumulation. PMID:19610030

  17. Mutations preventing expression of sup3 tRNASer nonsense suppressors of Schizosaccharomyces pombe.

    PubMed Central

    Pearson, D; Willis, I; Hottinger, H; Bell, J; Kumar, A; Leupold, U; Söll, D

    1985-01-01

    Suppression of nonsense codons in Schizosaccharomyces pombe by sup3-e tRNASerUGA or sup3-i tRNASerUAA is reduced or abolished by mutations within the suppressor locus. Twenty-five suppressor-inactive sup3-e genes and thirteen mutant sup3-i genes were isolated from S. pombe genomic clone banks by colony hybridization. Sequence analysis of these revertant alleles corroborates genetic evidence for mutational hotspots within the sup3 tRNA gene. Fifteen types of point mutations or insertions were found. Many of these replace bases which are highly or completely conserved in eucaryotic tRNA genes. Transcription of the altered sup3 genes in a Saccharomyces cerevisiae extract enabled the identification of mutations which affect the rate of 5'-end maturation or splicing of the tRNA precursors or both. A total of seven mutations were found which alter transcriptional efficiencies. Of these, five are located outside the internal transcription control regions. Images PMID:3921825

  18. MicroRNA-23a regulates 3T3-L1 adipocyte differentiation.

    PubMed

    Shen, Linyuan; Zhang, Yi; Du, Jingjing; Chen, Li; Luo, Jia; Li, Xuewei; Li, Mingzhou; Tang, Guoqing; Zhang, Shunhua; Zhu, Li

    2016-01-10

    MicroRNAs (miRNAs) are small, non-coding RNAs, which are involved in regulation of a variety of biological processes. Since previous studies regarding the role of miRNAs in the regulation of adipogenic differentiation have shown that miRNA-27a, one member of miRNA-23a∼27a∼24 cluster, could suppress adipogenesis. We now investigated whether miRNA-23a regulates adipogenic differentiation. In the present study, we showed that the expression of miRNA-23a is decreased during the process of adipogenic differentiation. Over-expression of miRNA-23a decreased lipid accumulation and triglyceride content in 3T3-L1 adipocytes. Our results also demonstrated that miRNA-23a decreases mRNA levels of adipocyte-specific genes involved in lipogenic transcription, fatty acid synthesis and fatty acid transport. These findings suggested miRNA-23a to be a new type of adipogenic depressor and to play an important role in regulating adipocyte differentiation. PMID:26415879

  19. Puerarin enhances adipocyte differentiation, adiponectin expression, and antioxidant response in 3T3-L1 cells.

    PubMed

    Lee, Ok-Hwan; Seo, Dong-Ho; Park, Cheon-Seok; Kim, Young-Cheul

    2010-01-01

    Puerarin, a major isoflavone glycoside from Kudzu root (Pueraria lobata), has been reported to exert antihyperglycemic and antioxidant effects and thus have pharmacological actions in the treatment of diabetes and cardiovascular diseases. We investigated the effects of puerarin on the changes of key gene expression associated with adipocyte differentiation and insulin sensitivity and link to cellular antioxidant response pathways. Puerarin treatment significantly enhanced differentiation of 3T3-L1 preadipocytes accompanying increased lipid accumulation and glucose-6-phosphate dehydrogenase (G6PDH) activity. At a molecular level, puerarin upregulated mRNA expression of peroxisome proliferator-activated receptor γ (PPARγ) and its target genes, an adipocyte-specific fatty acid binding protein (aP2) and GLUT4. Puerarin also caused a significant increase in mRNA level of adiponectin, an important insulin-sensitizing adipocytokine that is downregulated in insulin-resistant and diabetic states. In addition, treatment with puerarin was found to upregulate mRNA levels of G6PDH, glutathione reductase, and catalase, all of which are important for endogenous antioxidant responses. These data suggest that the hypoglycemic effects of puerarin can be attributed to the upregulation of PPARγ and its downstream target genes, GLUT4 and adiponectin expression, leading to increased glucose utilization. Puerarin may also be effective in preventing the rise of oxidative stress during adipocyte differentiation by increasing endogenous antioxidant responses. PMID:20806284

  20. Interleukin 1 amplifies receptor-mediated activation of phospholipase A2 in 3T3 fibroblasts.

    PubMed Central

    Burch, R M; Connor, J R; Axelrod, J

    1988-01-01

    Human recombinant interleukin 1 alpha (IL-1 alpha) and IL-1 beta stimulated prostaglandin E2 synthesis in 3T3 fibroblasts in a time- and concentration-dependent manner. Enhanced prostaglandin E2 synthesis after IL-1 treatment was apparent by 1 hr and continued to increase for at least 2 days. Half-maximal stimulation occurred at 0.5 pM IL-1 alpha or IL-1 beta, and both interleukins were equally effective, with maximal stimulation occurring in response to 5-10 pM IL-1. In contrast to IL-1, bradykinin stimulation of prostaglandin E2 synthesis is rapid; its effect is maximal by 5 min. In cells that had been pretreated with IL-1 for 24 hr, prostaglandin E2 synthesis in response to bradykinin was amplified more than 10-fold. IL-1 also amplified the receptor-mediated formation of prostaglandin E2 by bombesin and thrombin. The lymphokine did not affect bradykinin receptor number or affinity. IL-1 treatment induced phospholipase A2 and cyclooxygenase but not phospholipase C or prostaglandin E isomerase. It also enhanced bradykinin-stimulated GTPase activity, suggesting possible induction of the GTP-binding regulatory protein coupled to the bradykinin receptor. Thus, IL-1 enhanced receptor-mediated release of prostaglandin E2 in response to bradykinin, bombesin, and thrombin by increasing the cellular levels of phospholipase A2, cyclooxygenase, and GTP-binding regulatory protein(s). PMID:2901097

  1. Complete genome sequence of Veillonella parvula type strain (Te3T)

    SciTech Connect

    Gronow, Sabine; Welnitz, Sabine; Lapidus, Alla L.; Nolan, Matt; Ivanova, N; Glavina Del Rio, Tijana; Copeland, A; Chen, Feng; Tice, Hope; Pitluck, Sam; Cheng, Jan-Fang; Saunders, Elizabeth H; Brettin, Thomas S; Han, Cliff; Detter, J. Chris; Bruce, David; Goodwin, Lynne A.; Land, Miriam L; Hauser, Loren John; Chang, Yun-Juan; Jeffries, Cynthia; Pati, Amrita; Mavromatis, K; Mikhailova, Natalia; Chen, Amy; Palaniappan, Krishna; Chain, Patrick S. G.; Rohde, Manfred; Goker, Markus; Bristow, James; Eisen, Jonathan; Markowitz, Victor; Hugenholtz, Philip; Kyrpides, Nikos C; Klenk, Hans-Peter; Lucas, Susan

    2010-01-01

    Veillonella parvula (Veillon and Zuber 1898) Pr vot 1933 is the type species of the genus Veillonella in the family Veillonellaceae within the order Clostridiales. The species V. parvula is of interest because it is frequently isolated from dental plaque in the human oral cavity and can cause opportunistic infections. The species is strictly anaerobic and grows as small cocci which usually occur in pairs. Veillonellae are characterized by their unusual metabolism which is centered on the activity of the enzyme methylmalonyl-CoA decarboxylase. Strain Te3T, the type strain of the species, was isolated from the human intestinal tract. Here we describe the features of this organism, together with the complete genome sequence, and annotation. This is the first complete genome sequence of a member of the large clostridial family Veillonellaceae, and the 2,132,142 bp long single replicon genome with its 1859 protein-coding and 61 RNA genes is part of the Genomic Encyclopedia of Bacteria and Archaea project.

  2. Which pulse sequence is optimal for myo-Inositol detection at 3T?

    PubMed Central

    Hancu, Ileana

    2010-01-01

    Optimized myo-Inositol (mI) detection is important for diagnosing and monitoring a multitude of pathological conditions of the brain. Simulations are presented in this work, performed to decide which pulse sequence has the most significant advantage in terms of improving repeatability and accuracy of mI measurements at 3T over the pulse sequence used typically in the clinic, a TE=35ms PRESS sequence. Five classes of pulse sequences, four previously suggested for optimized mI detection (a short TE PRESS, a Carr-Purcell PRESS sequence, an optimized STEAM sequence, an optimized zero quantum filter), and one optimized for mI detection in this work (a single quantum filter) were compared to a standard, TE=35ms pulse sequence. While limiting the SNR of an acquisition to the equivalent SNR of a spectrum acquired in 5min from a 8cc voxel, it was found through simulations that the most repeatable mI measurements would be obtained with a Carr-Purcell sequence. This sequence was implemented in a clinical scanner, and improved mI measurements were demonstrated in vivo. PMID:19006101

  3. Segmenting time-lapse phase contrast images of adjacent NIH 3T3 cells.

    PubMed

    Chalfoun, J; Kociolek, M; Dima, A; Halter, M; Cardone, A; Peskin, A; Bajcsy, P; Brady, M

    2013-01-01

    We present a new method for segmenting phase contrast images of NIH 3T3 fibroblast cells that is accurate even when cells are physically in contact with each other. The problem of segmentation, when cells are in contact, poses a challenge to the accurate automation of cell counting, tracking and lineage modelling in cell biology. The segmentation method presented in this paper consists of (1) background reconstruction to obtain noise-free foreground pixels and (2) incorporation of biological insight about dividing and nondividing cells into the segmentation process to achieve reliable separation of foreground pixels defined as pixels associated with individual cells. The segmentation results for a time-lapse image stack were compared against 238 manually segmented images (8219 cells) provided by experts, which we consider as reference data. We chose two metrics to measure the accuracy of segmentation: the 'Adjusted Rand Index' which compares similarities at a pixel level between masks resulting from manual and automated segmentation, and the 'Number of Cells per Field' (NCF) which compares the number of cells identified in the field by manual versus automated analysis. Our results show that the automated segmentation compared to manual segmentation has an average adjusted rand index of 0.96 (1 being a perfect match), with a standard deviation of 0.03, and an average difference of the two numbers of cells per field equal to 5.39% with a standard deviation of 4.6%. PMID:23126432

  4. Resting state BOLD functional connectivity at 3T: spin echo versus gradient echo EPI.

    PubMed

    Chiacchiaretta, Piero; Ferretti, Antonio

    2015-01-01

    Previous evidence showed that, due to refocusing of static dephasing effects around large vessels, spin-echo (SE) BOLD signals offer an increased linearity and promptness with respect to gradient-echo (GE) acquisition, even at low field. These characteristics suggest that, despite the reduced sensitivity, SE fMRI might also provide a potential benefit when investigating spontaneous fluctuations of brain activity. However, there are no reports on the application of spin-echo fMRI for connectivity studies at low field. In this study we compared resting state functional connectivity as measured with GE and SE EPI sequences at 3T. Main results showed that, within subject, the GE sensitivity is overall larger with respect to that of SE, but to a less extent than previously reported for activation studies. Noteworthy, the reduced sensitivity of SE was counterbalanced by a reduced inter-subject variability, resulting in comparable group statistical connectivity maps for the two sequences. Furthermore, the SE method performed better in the ventral portion of the default mode network, a region affected by signal dropout in standard GE acquisition. Future studies should clarify if these features of the SE BOLD signal can be beneficial to distinguish subtle variations of functional connectivity across different populations and/or treatments when vascular confounds or regions affected by signal dropout can be a critical issue. PMID:25749359

  5. Induction of Adipocyte Differentiation by Polybrominated Diphenyl Ethers (PBDEs) in 3T3-L1 Cells

    PubMed Central

    Tung, Emily W. Y.; Boudreau, Adèle; Wade, Michael G.; Atlas, Ella

    2014-01-01

    Polybrominated diphenyl ethers (PBDEs) are a class of brominated flame retardants that were extensively used in commercial products. PBDEs are ubiquitous environmental contaminants that are both lipophilic and bioaccumulative. Effects of PBDEs on adipogenesis were studied in the 3T3-L1 preadipocyte cell model in the presence and absence of a known adipogenic agent, dexamethasone (DEX). A PBDE mixture designed to mimic body burden of North Americans was tested, in addition to the technical mixture DE-71 and the individual congener BDE-47. The mixture, DE-71, and BDE-47 all induced adipocyte differentiation as assessed by markers for terminal differentiation [fatty acid binding protein 4 (aP2) and perilipin] and lipid accumulation. Characterization of the differentiation process in response to PBDEs indicated that adipogenesis induced by a minimally effective dose of DEX was enhanced by these PBDEs. Moreover, C/EBPα, PPARγ, and LXRα were induced late in the differentiation process. Taken together, these data indicate that adipocyte differentiation is induced by PBDEs; they act in the absence of glucocorticoid and enhance glucocorticoid-mediated adipogenesis. PMID:24722056

  6. Modulation of Osteogenesis in MC3T3-E1 Cells by Different Frequency Electrical Stimulation

    PubMed Central

    Wang, Yu; Cui, Haitao; Wu, Zhenxu; Wu, Naipeng; Wang, Zongliang; Chen, Xuesi; Wei, Yen; Zhang, Peibiao

    2016-01-01

    Electrical stimulation (ES) is therapeutic to many bone diseases, from promoting fracture regeneration to orthopedic intervention. The application of ES offers substantial therapeutic potential, while optimal ES parameters and the underlying mechanisms responsible for the positive clinical impact are poorly understood. In this study, we assembled an ES cell culture and monitoring device. Mc-3T3-E1 cells were subjected to different frequency to investigate the effect of osteogenesis. Cell proliferation, DNA synthesis, the mRNA levels of osteosis-related genes, the activity of alkaline phosphatase (ALP), and intracellular concentration of Ca2+ were thoroughly evaluated. We found that 100 Hz could up-regulate the mRNA levels of collagen I, collagen II and Runx2. On the contrary, ES could down-regulate the mRNA levels of osteopontin (OPN). ALP activity assay and Fast Blue RR salt stain showed that 100 Hz could accelerate cells differentiation. Compared to the control group, 100 Hz could promote cell proliferation. Furthermore, 1 Hz to 10 Hz could improve calcium deposition in the intracellular matrix. Overall, these results indicate that 100Hz ES exhibits superior potentialities in osteogenesis, which should be beneficial for the clinical applications of ES for the treatment of bone diseases. PMID:27149625

  7. Monitoring fractional anisotropy in developing rabbit brain using MR diffusion tensor imaging at 3T

    NASA Astrophysics Data System (ADS)

    Jao, Jo-Chi; Yang, Yu-Ting; Hsiao, Chia-Chi; Chen, Po-Chou

    2016-03-01

    The aim of this study was to investigate the factional anisotropy (FA) in various regions of developing rabbit brain using magnetic resonance diffusion tensor imaging (MR DTI) at 3 T. A whole-body clinical MR imaging (MRI) scanner with a 15-channel high resolution knee coil was used. An echo-planar-imaging (EPI)-DTI pulse sequence was performed. Five 5 week-old New Zealand white (NZW) rabbits underwent MRI once per week for 24 weeks. After scanning, FA maps were obtained. ROIs (regions of interests) in the frontal lobe, parietal & temporal lobe, and occipital lobe were measured. FA changes with time were evaluated with a linear regression analysis. The results show that the FA values in all lobes of the brain increased linearly with age. The ranking of FA values was FA(frontal lobe) < FA(parietal & temporal lobe) > FA(occipital lobe). There was significant difference (p < 0.05) among these lobes. FA values are associated with the nerve development and brain functions. The FA change rate could be a biomarker to monitor the brain development. Understanding the FA values of various lobes during development could provide helpful information to diagnosis the abnormal syndrome earlier and have a better treatment and prognosis. This study established a brain MR-DTI protocol for rabbits to investigate the brain anatomy during development using clinical MRI. This technique can be further applied to the pre-clinical diagnosis, treatment, prognosis and follow-up of brain lesions.

  8. Most short DNA molecules isolated from 3T3 cells are not nascent.

    PubMed Central

    Kowalski, J; Denhardt, D T

    1978-01-01

    The population of short DNA molecules (less than 10(3) nucleotides) in 3T3 cells has been studied using in vivo and in vitro pulse labeling techniques and in vitro end-labeling. There is a large number of molecules of less than 100 nucleotides present in equal numbers in both Go and S phase cells. In S phase cells, most of these molecules are not replicating intermediates because they do not become density-labeled after a moderate period of substitution of BrdUMP, although they are detected by end-labeling in vitro. This population includes the nascent Okazaki pieces that can be labeled in a short pulse with [3H]dThd or [3H]dTTP, however, these represent less than 10% of the total population. Alkaline hydrolysis of the molecules that had been end-labeled with 32P using [gamma32P]ATP and polynucleotide kinase did not reveal significant release of [32P] 2'(3'), 5' ribonucleoside diphosphates. PMID:724517

  9. Contributed Review: Nuclear magnetic resonance core analysis at 0.3 T

    SciTech Connect

    Mitchell, Jonathan Fordham, Edmund J.

    2014-11-15

    Nuclear magnetic resonance (NMR) provides a powerful toolbox for petrophysical characterization of reservoir core plugs and fluids in the laboratory. Previously, there has been considerable focus on low field magnet technology for well log calibration. Now there is renewed interest in the study of reservoir samples using stronger magnets to complement these standard NMR measurements. Here, the capabilities of an imaging magnet with a field strength of 0.3 T (corresponding to 12.9 MHz for proton) are reviewed in the context of reservoir core analysis. Quantitative estimates of porosity (saturation) and pore size distributions are obtained under favorable conditions (e.g., in carbonates), with the added advantage of multidimensional imaging, detection of lower gyromagnetic ratio nuclei, and short probe recovery times that make the system suitable for shale studies. Intermediate field instruments provide quantitative porosity maps of rock plugs that cannot be obtained using high field medical scanners due to the field-dependent susceptibility contrast in the porous medium. Example data are presented that highlight the potential applications of an intermediate field imaging instrument as a complement to low field instruments in core analysis and for materials science studies in general.

  10. Cytoplasmic pH influences cytoplasmic calcium in MC3T3-E1 osteoblast cells

    NASA Technical Reports Server (NTRS)

    Lin, H. S.; Hughes-Fulford, M.; Kumegawa, M.; Pitts, A. C.; Snowdowne, K. W.

    1993-01-01

    We found that the cytoplasmic concentration of calcium (Cai) of MC3T3-E1 osteoblasts was influenced by the type of pH buffer we used in the perfusing medium, suggesting that intracellular pH (pHi) might influence Cai. To study this effect, the Cai and pHi were monitored as we applied various experimental conditions known to change pHi. Exposure to NH4Cl caused a transient increase in both pHi and Cai without a change in extracellular pH (pHo). Decreasing pHo and pHi by lowering the bicarbonate concentration of the medium decreased Cai, and increasing pHi by the removal of 5% CO2 increased Cai. Clamping pHi to known values with 10 microM nigericin, a potassium proton ionophore, also influenced Cai: acid pHi lowered Cai, whereas alkaline pHi increased it. The rise in Cai appears to be very sensitive to the extracellular concentration of calcium, suggesting the existence of a pH-sensitive calcium influx mechanism. We conclude that physiologic changes in pH could modulate Cai by controlling the influx of calcium ions and could change the time course of the Cai transient associated with hormonal activation.

  11. Using High Spatial Resolution to Improve BOLD fMRI Detection at 3T

    PubMed Central

    Claise, Béatrice; Jean, Betty

    2015-01-01

    For different functional magnetic resonance imaging experiments using blood oxygenation level-dependent (BOLD) contrast, the acquisition of T2*-weighted scans at a high spatial resolution may be advantageous in terms of time-course signal-to-noise ratio and of BOLD sensitivity when the regions are prone to susceptibility artifacts. In this study, we explore this solution by examining how spatial resolution influences activations elicited when appetizing food pictures are viewed. Twenty subjects were imaged at 3 T with two different voxel volumes, 3.4 μl and 27 μl. Despite the diminution of brain coverage, we found that high-resolution acquisition led to a better detection of activations. Though known to suffer to different degrees from susceptibility artifacts, the activations detected by high spatial resolution were notably consistent with those reported in published activation likelihood estimation meta-analyses, corresponding to taste-responsive regions. Furthermore, these regions were found activated bilaterally, in contrast with previous findings. Both the reduction of partial volume effect, which improves BOLD contrast, and the mitigation of susceptibility artifact, which boosts the signal to noise ratio in certain regions, explained the better detection noted with high resolution. The present study provides further evidences that high spatial resolution is a valuable solution for human BOLD fMRI, especially for studying food-related stimuli. PMID:26550990

  12. Improved localisation for 2-hydroxyglutarate detection at 3T using long-TE semi-LASER

    PubMed Central

    Berrington, Adam; Voets, Natalie L.; Plaha, Puneet; Larkin, Sarah J.; Mccullagh, James; Stacey, Richard; Yildirim, Muhammed; Schofield, Christopher J.; Jezzard, Peter; Cadoux-Hudson, Tom; Ansorge, Olaf; Emir, Uzay E.

    2016-01-01

    2-hydroxyglutarate (2-HG) has emerged as a biomarker of tumour cell IDH mutations that may enable the differential diagnosis of glioma patients. At 3 Tesla, detection of 2-HG with magnetic resonance spectroscopy is challenging because of metabolite signal overlap and a spectral pattern modulated by slice selection and chemical shift displacement. Using density matrix simulations and phantom experiments, an optimised semi-LASER scheme (TE = 110 ms) improves localisation of the 2-HG spin system considerably compared to an existing PRESS sequence. This results in a visible 2-HG peak in the in vivo spectra at 1.9 ppm in the majority of IDH mutated tumours. Detected concentrations of 2-HG were similar using both sequences, although the use of semi-LASER generated narrower confidence intervals. Signal overlap with glutamate and glutamine, as measured by pairwise fitting correlation was reduced. Lactate was readily detectable across glioma patients using the method presented here (mean CLRB: (10±2)%). Together with more robust 2-HG detection, long TE semi-LASER offers the potential to investigate tumour metabolism and stratify patients in vivo at 3T. PMID:27547821

  13. WEHI-3 cells inhibit adipocyte differentiation in 3T3-L1 cells.

    PubMed

    Lai, Jing; Liu, Gexiu; Yan, Guoyao; He, Dongmei; Zhou, Ying; Chen, Shengting

    2015-06-26

    By investigating the anti-adipogenic effects of WEHI-3 cells - a murine acute myelomonocytic leukemia cell line - we sought to improve the efficiency of hematopoietic stem cell transplantation (HSCT). Analysis of Oil Red O staining and the expression of adipogenic genes, including PPARγ, C/EBPα, FAS and LPL, indicated that WEHI-3 cells significantly inhibited 3T3-L1 mouse preadipocyte cells from differentiating into adipocytes. In vivo, fat vacuoles in mice injected with WEHI-3 cells were also remarkably reduced in the murine bone marrow pimelosis model. Moreover, the key gene in the Rho signaling pathway, ROCKII, and the key gene in the Wnt signaling pathway, β-catenin, were both upregulated compared with the control group. siRNA-mediated knockdown of ROCKII and β-catenin reversed these WEHI-3-mediated anti-adipogenic effects. Taken together, these data suggest that WEHI-3 cells exert anti-adipogenic effects and that both ROCKII and β-catenin are involved in this process. PMID:25911323

  14. Antidiabetic thiazolidinediones inhibit leptin (ob) gene expression in 3T3-L1 adipocytes.

    PubMed Central

    Kallen, C B; Lazar, M A

    1996-01-01

    Lack of leptin (ob) protein causes obesity in mice. The leptin gene product is important for normal regulation of appetite and metabolic rate and is produced exclusively by adipocytes. Leptin mRNA was induced during the adipose conversion of 3T3-L1 cells, which are useful for studying adipocyte differentiation and function under controlled conditions. We studied leptin regulation by antidiabetic thiazolidinedione compounds, which are ligands for the adipocyte-specific nuclear receptor peroxisome proliferator-activated receptor gamma (PPARgamma) that regulates the transcription of other adipocyte-specific genes. Remarkably, leptin gene expression was dramatically repressed within a few hours after thiazolidinedione treatment. The ED50 for inhibition of leptin expression by the thiazolidinedione BRL49653 was between 5 and 50 nM, similar to its Kd for binding to PPARgamma. The relatively weak, nonthiazolidinedione PPAR activator WY 14,643 also inhibited leptin expression, but was approximately 1000 times less potent than BRL49653. These results indicate that antidiabetic thiazolidinediones down-regulate leptin gene expression with potencies that correlate with their abilities to bind and activate PPARgamma. Images Fig. 1 Fig. 2 Fig. 3 Fig. 4 PMID:8650171

  15. MRBrainS Challenge: Online Evaluation Framework for Brain Image Segmentation in 3T MRI Scans

    PubMed Central

    Mendrik, Adriënne M.; Vincken, Koen L.; Kuijf, Hugo J.; Breeuwer, Marcel; Bouvy, Willem H.; de Bresser, Jeroen; Alansary, Amir; de Bruijne, Marleen; Carass, Aaron; El-Baz, Ayman; Jog, Amod; Katyal, Ranveer; Khan, Ali R.; van der Lijn, Fedde; Mahmood, Qaiser; Mukherjee, Ryan; van Opbroek, Annegreet; Paneri, Sahil; Pereira, Sérgio; Rajchl, Martin; Sarikaya, Duygu; Smedby, Örjan; Silva, Carlos A.; Vrooman, Henri A.; Vyas, Saurabh; Wang, Chunliang; Zhao, Liang; Biessels, Geert Jan; Viergever, Max A.

    2015-01-01

    Many methods have been proposed for tissue segmentation in brain MRI scans. The multitude of methods proposed complicates the choice of one method above others. We have therefore established the MRBrainS online evaluation framework for evaluating (semi)automatic algorithms that segment gray matter (GM), white matter (WM), and cerebrospinal fluid (CSF) on 3T brain MRI scans of elderly subjects (65–80 y). Participants apply their algorithms to the provided data, after which their results are evaluated and ranked. Full manual segmentations of GM, WM, and CSF are available for all scans and used as the reference standard. Five datasets are provided for training and fifteen for testing. The evaluated methods are ranked based on their overall performance to segment GM, WM, and CSF and evaluated using three evaluation metrics (Dice, H95, and AVD) and the results are published on the MRBrainS13 website. We present the results of eleven segmentation algorithms that participated in the MRBrainS13 challenge workshop at MICCAI, where the framework was launched, and three commonly used freeware packages: FreeSurfer, FSL, and SPM. The MRBrainS evaluation framework provides an objective and direct comparison of all evaluated algorithms and can aid in selecting the best performing method for the segmentation goal at hand. PMID:26759553

  16. Hippocampal blood flow in normal aging measured with arterial spin labeling at 3T

    PubMed Central

    Rusinek, Henry; Brys, Miroslaw; Glodzik, Lidia; Switalski, Remigiusz; Tsui, Wai-Hon; Haas, Francois; Mcgorty, Kellyanne; Chen, Qun; de Leon, Mony J.

    2011-01-01

    Due to methodological difficulties related to the small size, variable distribution of hippocampal arteries, and the location of the hippocampus in the proximity of middle cranial fossa, little is known about hippocampal blood flow (HBF). We have tested the utility of a pulsed ASL sequence based on multi-shot TrueFISP to measure HBF in 34 normal volunteers (17 women, 17 men, 26-92 years old). Flow sensitivity to a mild hypercapnic challenge was also examined. Coregistered 3D MPRAGE sequence was used to eliminate from hippocampal and cortical regions of interest all voxel with <75% of gray matter. Large blood vessels were also excluded. HBF in normal volunteers averaged 61.2±9.0 ml/(100g min). There was no statistically significant age or gender effect. Under a mild hypercapnia challenge (end tidal CO2 pressure increase of 6.8±1.9 mg Hg over the baseline), HBF response was 14.1±10.8 ml/(100g min), whereas cortical gray matter flow increased by 18.0±12.2 ml/(100g min). Flow response among women was significantly larger than in the men. The average absolute difference between two successive HBF measures was 3.6 ml/(100g min), or 5.4%. The 3T TrueFISP ASL method offers a HBF measurement strategy that combines good spatial resolution, sensitivity, and minimal image distortions. PMID:20939094

  17. In vivo detection of citrate in brain tumors by 1H MRS at 3T

    PubMed Central

    Choi, Changho; Ganji, Sandeep K.; Madan, Akshay; Hulsey, Keith M.; An, Zhongxu; Zhang, Song; Pinho, Marco C.; DeBerardinis, Ralph J.; Bachoo, Robert M.; Maher, Elizabeth A.

    2014-01-01

    Purpose To test whether the citrate is elevated in adult patients with gliomas using 1H MRS at 3T in vivo. Methods Thirty-four adult patients were enrolled in the study, including 6 subjects with glioblastomas, 8 subjects with astrocytomas (5 WHO grade-3 and 3 grade-2), and 20 subjects with oligodendrogliomas (5 grade-3 and 15 grade-2). Five healthy volunteers were studied for baseline citrate data. Single-voxel localized spectra were collected with PRESS TE = 35 and 97 ms and analyzed with LCModel using numerically calculated basis spectra that include the effects of the PRESS radio-frequency and gradient pulses. Results Citrate was not measurable by MRS in healthy brain, but was detected in tumor patients at both echo times. The citrate concentration was estimated to be as high as 1.8 mM with reference to water at 42 M, with CRLB as low as 5%. The mean citrate level was 0.7±0.4 mM (mean±SD, n=32) with median CRLB of ~12%. No correlation was identified between citrate concentration and tumor grade or histological type. Conclusion Citrate was increased in the majority of gliomas in adult patients. The elevated citrate in our data indicates an altered metabolic state of tumor relative to healthy brain. PMID:24123337

  18. Aldosterone perturbs adiponectin and PAI-1 expression and secretion in 3T3-L1 adipocytes.

    PubMed

    Li, P; Zhang, X-N; Pan, C-M; Sun, F; Zhu, D-L; Song, H-D; Chen, M-D

    2011-06-01

    Aldosterone is considered as a new cardiovascular risk factor that plays an important role in metabolic syndrome; however, the underlying mechanism of these effects is not clear. Hypoadiponectinemia and elevated circulating concentration of plasminogen activator inhibitor-1 (PAI-1) are causally associated with obesity-related insulin resistance and cardiovascular disease. The aim of the present study is to investigate the effect of aldosterone on the production of adiponectin and PAI-1 in 3T3-L1 adipocytes. Northern and Western blot analyses revealed that aldosterone treatment inhibited adiponectin mRNA expression and secretion and simultaneously enhanced PAI-1 mRNA expression and secretion in a time- and dose-dependent manner. Rosiglitazone did not prevent aldosterone's effect on adiponectin or PAI-1 expression. In contrast, tumor necrosis factor (TNF)-α produced dramatic synergistic effects on adiponectin and PAI-1 expression when added together with aldosterone. Furthermore, the effects of aldosterone on adiponectin and PAI-1 expression appear to be mediated through glucocorticoid receptor (GR) but not mineralocorticoid receptor (MR). These results suggest that the effects of aldosterone on adiponectin and PAI-1 production are one of the underlying mechanisms linking it to insulin resistance, metabolic syndrome and cardiovascular disease. PMID:21667402

  19. Sphingosine-1-phosphate inhibits the adipogenic differentiation of 3T3-L1 preadipocytes.

    PubMed

    Moon, Myung-Hee; Jeong, Jae-Kyo; Lee, You-Jin; Seol, Jae-Won; Park, Sang-Youel

    2014-10-01

    Sphingosine-1-phosphate (S1P) is a pluripotent lipid mediator that transmits signals through G-protein-coupled receptors to control diverse biological processes. The novel biological activity of S1P in the adipogenesis of 3T3-L1 preadipocytes was identified in the present study. S1P significantly decreased lipid accumulation in maturing preadipocytes in a dose‑dependent manner. In order to understand the anti‑adipogenic effects of S1P, preadipocytes were treated with S1P, and the change in the expression of several adipogenic transcription factors and enzymes was investigated using quantitative RT-PCR. S1P downregulated the transcriptional levels of the peroxisome proliferator-activated receptor γ, CCAAT/enhancer binding proteins and adiponectin, which are markers of adipogenic differentiation. The effects of S1P on the levels of mitogen‑activated protein kinase (MAPK) signals in preadipocytes were also investigated. The activation of JNK and p38 were downregulated by S1P treatment in human preadipocytes. In conclusion, the results of this study suggest that S1P alters fat mass by directly affecting adipogenesis. This is mediated by the downregulation of adipogenic transcription factors and by inactivation of the JNK and p38 MAPK pathways. Thus, selective targeting of the S1P receptors and sphingosine kinases may have clinical applications for the treatment of obesity. PMID:25050633

  20. MC3T3-E1 Cell Response to Pure Titanium, Zirconia and Nano-Hydroxyapatite

    NASA Astrophysics Data System (ADS)

    Lee, Dong-Hwan; Han, Jung-Suk; Yang, Jae-Ho; Lee, Jai-Bong; Kim, Dae-Joon

    Titanium, zirconia and HAp were known as good biocompatible materials for tissue engineering. Osteblastic cell response is influence by the surface topography of material and its chemical composition as well. To evaluate the influence of different chemical compositions on osteoblast-like cells the specimens were polished until they have almost identical surface roughness. The commercially pure titanium, zirconia/alumina composite and nano-sized hydroxyapatite (HAp) specimens synthesized by hydrothermal method were used to evaluate the cell attachment, proliferation and differentiation. Confocal laser microscopy was used measurement of surface roughness, and flourescence microscopy and SEM were used to evaluate initial cell attachment and morphology after 3 hours. MTS assay was performed for cell proliferation after 1, 3, 7 days and ALP assay was used for cell differentiation after 7, 10, 14 days of cell culture period. Surface topography of nano-HAp specimen was almost identical compared with those of titanium and zirconia specimen. Under this condition, proliferation and differentiation of MC3T3-E1 cells was not significantly different with those on titanium and zirconia specimen. However, cells on Nano-HAp specimen showed quicker and more active cellular reaction for attachment when measured by the expression of adhesion proteins through confocal laser microscopy. The results suggested that the new nano-sized HAp can be applied as a suitable material for skeletal tissue engineering.

  1. Automatic tuned MRI RF coil for multinuclear imaging of small animals at 3T.

    PubMed

    Muftuler, L Tugan; Gulsen, Gultekin; Sezen, Kumsal D; Nalcioglu, Orhan

    2002-03-01

    We have developed an MRI RF coil whose tuning can be adjusted automatically between 120 and 128 MHz for sequential spectroscopic imaging of hydrogen and fluorine nuclei at field strength 3 T. Variable capacitance (varactor) diodes were placed on each rung of an eight-leg low-pass birdcage coil to change the tuning frequency of the coil. The diode junction capacitance can be controlled by the amount of applied reverse bias voltage. Impedance matching was also done automatically by another pair of varactor diodes to obtain the maximum SNR at each frequency. The same bias voltage was applied to the tuning varactors on all rungs to avoid perturbations in the coil. A network analyzer was used to monitor matching and tuning of the coil. A Pentium PC controlled the analyzer through the GPIB bus. A code written in LABVIEW was used to communicate with the network analyzer and adjust the bias voltages of the varactors via D/A converters. Serially programmed D/A converter devices were used to apply the bias voltages to the varactors. Isolation amplifiers were used together with RF choke inductors to provide isolation between the RF coil and the DC bias lines. We acquired proton and fluorine images sequentially from a multicompartment phantom using the designed coil. Good matching and tuning were obtained at both resonance frequencies. The tuning and matching of the coil were changed from one resonance frequency to the other within 60 s. PMID:11945031

  2. Platelet-derived growth factor (PDGF) stimulates glycogen synthase activity in 3T3 cells

    SciTech Connect

    Chan, C.P.; Bowen-Pope, D.F.; Ross, R.; Krebs, E.G.

    1986-05-01

    Hormonal regulation of glycogen synthase, an enzyme that can be phosphorylated on multiple sites, is often associated with changes in its phosphorylation state. Enzyme activation is conventionally monitored by determining the synthase activity ratio ((activity in the absence of glucose 6-P)/(activity in the presence of glucose 6-P)). Insulin causes an activation of glycogen synthase with a concomitant decrease in its phosphate content. In a previous report, the authors showed that epidermal growth factor (EGF) increases the glycogen synthase activity ratio in Swiss 3T3 cells. The time and dose-dependency of this response was similar to that of insulin. Their recent results indicate that PDGF also stimulates glycogen synthase activity. Enzyme activation was maximal after 30 min. of incubation with PDGF; the time course observed was very similar to that with insulin and EGF. At 1 ng/ml (0.03nM), PDGF caused a maximal stimulation of 4-fold in synthase activity ratio. Half-maximal stimulation was observed at 0.2 ng/ml (6 pM). The time course of changes in enzyme activity ratio closely followed that of /sup 125/I-PDGF binding. The authors data suggest that PDGF, as well as EFG and insulin, may be important in regulating glycogen synthesis through phosphorylation/dephosphorylation mechanisms.

  3. Hindered Diffusion of Inert Tracer Particles in the Cytoplasm of Mouse 3T3 Cells

    NASA Astrophysics Data System (ADS)

    Luby-Phelps, Katherine; Castle, Philip E.; Lansing Taylor, D.; Lanni, Frederick

    1987-07-01

    Using fluorescence recovery after photobleaching, we have studied the diffusion of fluorescein-labeled, size-fractionated Ficoll in the cytoplasmic space of living Swiss 3T3 cells as a probe of the physical chemical properties of cytoplasm. The results reported here corroborate and extend the results of earlier experiments with fluorescein-labeled, size-fractionated dextran: diffusion of nonbinding particles in cytoplasm is hindered in a size-dependent manner. Extrapolation of the data suggests that particles larger than 260 angstrom in radius may be completely nondiffusible in the cytoplasmic space. In contrast, diffusion of Ficoll in protein solutions of concentration comparable to the range reported for cytoplasm is not hindered in a size-dependent manner. Although we cannot at present distinguish among several physical chemical models for the organization of cytoplasm, these results make it clear that cytoplasm possesses some sort of higher-order intermolecular interactions (structure) not found in simple aqueous protein solutions, even at high concentration. These results also suggest that, for native cytoplasmic particles whose smallest radial dimension approaches 260 angstrom, size may be as important a determinant of cytoplasmic diffusibility as binding specificity. This would include most endosomes, polyribosomes, and the larger multienzyme complexes.

  4. Nobiletin enhances differentiation and lipolysis of 3T3-L1 adipocytes

    SciTech Connect

    Saito, Takeshi; Abe, Daigo; Sekiya, Keizo . E-mail: ksekiya@affrc.go.jp

    2007-06-01

    Nobiletin is a polymethoxylated flavone found in certain citrus fruits. Here we demonstrate that nobiletin enhance differentiation of 3T3-L1 preadipocytes. Nobiletin dose-dependently increased accumulation of lipid droplets in adipocytes. Quantitative RT-PCR analyses indicated that nobiletin increased the expression of genes critical for acquisition of the adipocyte phenotype. Some of them were known peroxisome proliferator activated receptor {gamma} (PPAR{gamma}) targets and PPAR{gamma} itself, however, nobiletin did not exhibit PPAR{gamma} ligand activity. We observed the expression of CCAAT/enhancer binding protein {beta} (C/EBP{beta}), a transcription factor for PPAR{gamma}, was increased by nobiletin. The activation of cAMP-responsive element binding protein (CREB) and extracellular signal-regulated kinase (ERK), which play important roles in C/EBP{beta} expression were also potentiated by nobiletin. Furthermore, nobiletin stimulated lipolysis in differentiated adipocytes, which is known to be stimulated by cAMP pathway. These results suggested that nobiletin enhanced both differentiation and lipolysis of adipocyte through activation of signaling cascades mediated by cAMP/CREB.

  5. Manganese superoxide dismutase knock-down in 3T3-L1 preadipocytes impairs subsequent adipogenesis.

    PubMed

    Krautbauer, Sabrina; Eisinger, Kristina; Hader, Yvonne; Neumeier, Markus; Buechler, Christa

    2014-08-01

    Adipogenesis is associated with the upregulation of the antioxidative enzyme manganese superoxide dismutase (MnSOD) suggesting a vital function of this enzyme in adipocyte maturation. In the current work, MnSOD was knocked-down with small-interference RNA in preadipocytes to study its role in adipocyte differentiation. In mature adipocytes differentiated from these cells, proteins characteristic for mature adipocytes, which are strongly induced in late adipogenesis like adiponectin and fatty acid-binding protein 4, are markedly reduced. Triglycerides begin to accumulate after about 6 days of the induction of adipogenesis, and are strongly diminished in cells with low MnSOD. Proteins upregulated early during differentiation, like fatty acid synthase and cytochrome C oxidase-4, are not altered. Cell viability, insulin-mediated phosphorylation of Akt, antioxidative capacity (AOC), superoxide levels, and heme oxygenase 1 with the latter being induced upon oxidative stress are not affected. L-Buthionine-(S,R)-sulfoximine (BSO) depletes glutathione and modestly lowers AOC of mature adipocytes. Addition of BSO to 3T3-L1 cells 3 days after the initiation of differentiation impairs triglyceride accumulation and expression of proteins induced in late adipogenesis. Of note, proteins that increased early during adipogenesis are also diminished, suggesting that BSO causes de-differentiation of these cells. Preadipocyte proliferation is not considerably affected by low MnSOD and BSO. These data suggest that glutathione and MnSOD are essential for adipogenesis. PMID:24740755

  6. A 64-channel 3T array coil for accelerated brain MRI

    PubMed Central

    Keil, Boris; Blau, James N.; Biber, Stephan; Hoecht, Philipp; Tountcheva, Veneta; Setsompop, Kawin; Triantafyllou, Christina; Wald, Lawrence L.

    2012-01-01

    A 64-channel brain array coil was developed and compared to a 32-channel array constructed with the same coil former geometry in order to precisely isolate the benefit of the two-fold increase in array coil elements. The constructed coils were developed for a standard clinical 3T MRI scanner and used a contoured head-shape curved former around the occipital pole and tapered in at the neck to both improve sensitivity and patient comfort. Additionally, the design is a compact, split-former design intended for robust daily use. Signal-to-noise ratio (SNR) and noise amplification (G-factor) for parallel imaging were quantitatively evaluated in human imaging and compared to a size and shape-matched 32-channel array coil. For unaccelerated imaging, the 64-channel array provided similar SNR in the brain center to the 32-channel array and 1.3-fold more SNR in the brain cortex. Reduced noise amplification during highly parallel imaging of the 64-channel array provided the ability to accelerate at approximately one unit higher at a given noise amplification compared to the sized-matched 32-channel array. For example, with a 4-fold acceleration rate, the central brain and cortical SNR of the 64-channel array was 1.2 and 1.4-fold higher, respectively, compared to the 32-channel array. The characteristics of the coil are demonstrated in accelerated brain imaging. PMID:22851312

  7. Protein kinase C activation by anthracyclines in Swiss 3T3 cells.

    PubMed

    Lanzi, C; Gambetta, R A; Perego, P; Banfi, P; Franzi, A; Guazzoni, L; Zunino, F

    1991-01-01

    The effects of the anti-cancer anthracyclines doxorubicin and daunorubicin on the activity of protein kinase C (PKC) were examined in intact Swiss 3T3 cells. The 2 drugs stimulated the phosphorylation of an 80K phosphoprotein found to be identical to that generated in response to the PKC activator 12-O-tetradecanoylphorbol-13-acetate as indicated by gel electrophoresis and peptide mapping. The effect of doxorubicin was dose-dependent in the range 10(-5) to 10(-3) M and was not associated with a detectable translocation of PKC activity from cytosol to the cell membrane. Doxorubicin and daunorubicin were found to increase the incorporation of phosphate into phosphatidic acid, phosphatidylinositol 4-monophosphate and phosphatidyl inositol 4,5-bisphosphate. In addition, the anthracyclines induced a rise in inositol phosphates, thus indicating a stimulation of the breakdown of phosphoinositides. These data are consistent with an indirect mechanism of PKC activation by anthracyclines. We propose that diacylglycerol, which is derived from the hydrolysis of phospholipids, (including the phosphoinositides), by activation of phospholipases, could mediate PKC activation. The described effects, involving cell-signal-transducing pathways, emphasize a new aspect of the cellular actions of these anti-tumor agents. PMID:1845961

  8. Transformation of NIH 3T3 cells by cotransfection with c-src and nuclear oncogenes.

    PubMed Central

    Shalloway, D; Johnson, P J; Freed, E O; Coulter, D; Flood, W A

    1987-01-01

    pp60c-src, the cellular homolog of the Rous sarcoma virus transforming protein, does not completely transform cells even when present at high levels, but has been shown to be involved in polyomavirus-induced transformation when activated by polyomavirus middle T (pmt)-antigen binding. Here we show that cotransfection, but not solo transfection, of expression plasmids for c-src and either adenovirus E1A, v-myc, c-myc, or the 5' half of polyomavirus large T (pltN) antigen into NIH 3T3 cells induces anchorage-independent growth, enhanced focus formation, and, for pltN cotransfection, tumorigenicity in adult NFS mice. Enhancement of transformation was not observed with polyomavirus small t (pst) antigen. Cotransfection of c-src with pltN induced modification of pp60c-src that altered its electrophoretic mobility and in vivo phosphorylation state and stimulated its in vitro kinase activity. Similar alterations were not seen after c-src-E1A cotransfection, suggesting that at least two different mechanisms of enhancement are involved. Images PMID:2446117

  9. In vivo 13 carbon metabolic imaging at 3T with hyperpolarized 13C-1-pyruvate.

    PubMed

    Kohler, S J; Yen, Y; Wolber, J; Chen, A P; Albers, M J; Bok, R; Zhang, V; Tropp, J; Nelson, S; Vigneron, D B; Kurhanewicz, J; Hurd, R E

    2007-07-01

    We present for the first time dynamic spectra and spectroscopic images acquired in normal rats at 3T following the injection of (13)C-1-pyruvate that was hyperpolarized by the dynamic nuclear polarization (DNP) method. Spectroscopic sampling was optimized for signal-to-noise ratio (SNR) and for spectral resolution of (13)C-1-pyruvate and its metabolic products (13)C-1-alanine, (13)C-1-lactate, and (13)C-bicarbonate. Dynamic spectra in rats were collected with a temporal resolution of 3 s from a 90-mm axial slab using a dual (1)H-(13)C quadrature birdcage coil to observe the combined effects of metabolism, flow, and T(1) relaxation. In separate experiments, spectroscopic imaging data were obtained during a 17-s acquisition of a 20-mm axial slice centered on the rat kidney region to provide information on the spatial distribution of the metabolites. Conversion of pyruvate to lactate, alanine, and bicarbonate occurred within a minute of injection. Alanine was observed primarily in skeletal muscle and liver, while pyruvate, lactate, and bicarbonate concentrations were relatively high in the vasculature and kidneys. In contrast to earlier work at 1.5 T, bicarbonate was routinely observed in skeletal muscle as well as the kidney and vasculature. PMID:17659629

  10. Hsp90 chaperones PPARγ and regulates differentiation and survival of 3T3-L1 adipocytes

    PubMed Central

    Nguyen, M T; Csermely, P; Sőti, C

    2013-01-01

    Adipose tissue dysregulation has a major role in various human diseases. The peroxisome proliferator-activated receptor-γ (PPARγ) is a key regulator of adipocyte differentiation and function, as well as a target of insulin-sensitizing drugs. The Hsp90 chaperone stabilizes a diverse set of signaling ‘client' proteins, thereby regulates various biological processes. Here we report a novel role for Hsp90 in controlling PPARγ stability and cellular differentiation. Specifically, we show that the Hsp90 inhibitors geldanamycin and novobiocin efficiently impede the differentiation of murine 3T3-L1 preadipocytes. Geldanamycin at higher concentrations also inhibits the survival of both developing and mature adipocytes, respectively. Further, Hsp90 inhibition disrupts an Hsp90-PPARγ complex, leads to the destabilization and proteasomal degradation of PPARγ, and inhibits the expression of PPARγ target genes, identifying PPARγ as an Hsp90 client. A similar destabilization of PPARγ and a halt of adipogenesis also occur in response to protein denaturing stresses caused by a single transient heat-shock or proteasome inhibition. Recovery from stress restores PPARγ stability and adipocyte differentiation. Thus, our findings reveal Hsp90 as a critical stress-responsive regulator of adipocyte biology and offer a potential therapeutic target in obesity and the metabolic syndrome. PMID:24096869

  11. In vivo conductivity imaging of canine male pelvis using a 3T MREIT system

    NASA Astrophysics Data System (ADS)

    Kim, H. J.; Jeong, W. C.; Kim, Y. T.; Minhas, A. S.; Lee, T. H.; Lim, C. Y.; Park, H. M.; Seo, J. K.; Woo, E. J.

    2010-04-01

    The prostate is an imaging area of growing concern related with aging. Prostate cancer and benign prostatic hyperplasia are the most common diseases and significant cause of death for elderly men. Hence, the conductivity imaging of the male pelvis is a challenging task with a clinical significance. In this study, we performed in vivo MREIT imaging experiments of the canine male pelvis using a 3T MRI scanner. Adopting carbon-hydrogel electrodes and a multi-echo pulse sequence, we could inject as much as 10 mA current in a form of 51 ms pulse into the pelvis. Collecting magnetic flux density data inside the pelvis subject to multiple injection currents, we reconstructed cross-sectional conductivity images using a MREIT software package CoReHA. Scaled conductivity images of the prostate show a clear contrast between the central and peripheral zones which are related with prostate diseases including cancer and benign prostatic hyperplasia. In our future work, we will focus on prostate cancer model animal experiments.

  12. Differentiation of the insulin-sensitive glucose transporter in 3T3-L1 adipocytes

    SciTech Connect

    Frost, S.C.; Baly, D.L.; Cushman, S.W.; Lane, M.D.; Simpson, I.A.

    1986-05-01

    3T3-L1 fibroblasts differentiate in culture to resemble adipocytes both morphologically and biochemically. Insulin-sensitive glucose transport, as measured by 2-deoxy-(1-/sup 14/C)- glucose uptake in the undifferentiated cell is small (2X). In contrast, the rate of glucose transport in fully differentiated cells is elevated 15-fold over basal in the presence of insulin. To determine if this is due to an increase in the number of transporters/cell or accessibility to the transporters, the number of transporters was measured in subcellular fractions over differentiation using a /sup 3/H-cytochalasin B binding assay. The increase in the rate of insulin-sensitive glucose transport directly parallels an increase in the number of transporters which reside in an insulin-responsive intracellular compartment. This observation was confirmed by identifying the transporters by immunoblotting using an antibody generated against the human erythrocyte transporter. The molecular weight of this transporter increases over differentiation from a single band of 40kDa to a heterogeneous triplet of 40, 44 and 48kDa. These data suggest that the transporter undergoes differential processing and that the functional, insulin-responsive transporter may be different from the insulin-insensitive (basal) transporter.

  13. (-)-Ternatin inhibits adipogenesis and lipid metabolism in 3T3-L1 cells.

    PubMed

    Ito, Masahiko; Ito, Junko; Kitazawa, Hidefumi; Shimamura, Ken; Fukami, Takehiro; Tokita, Shigeru; Shimokawa, Kenichiro; Yamada, Kaoru; Kanatani, Akio; Uemura, Daisuke

    2009-06-01

    (-)-Ternatin, a highly N-methylated cyclic peptide, inhibits fat accumulation in 3T3-L1 cells and reduces fat mass in mice. However, the mechanism for its anti-adipogenic effect has remained unknown. To examine the mechanism used by (-)-ternatin to inhibit adipocyte differentiation, we examined the effects of (-)-ternatin and [l-Ala(4)]ternatin, an inactive analog of (-)-ternatin, on the expression of adipocyte markers and lipogenic enzymes. We found that (-)-ternatin potently reduced mRNA expression of several adipocyte markers in a dose-dependent manner, whereas [l-Ala(4)]ternatin showed no effects. At the immediate early phase, (-)-ternatin, but not [l-Ala(4)]ternatin, reduced the expression of Srebp1c, Fas, Acc2 and C/EBP-alpha while showing no effects on C/EBP-beta and C/EBP-delta. These results suggest that (-)-ternatin affects the mid-to late differentiation stages of adipocytes. Consistent with the decreased expression of lipogenic enzymes, (-)-ternatin potently inhibited triglyceride synthesis. Intriguingly, (-)-ternatin also inhibited triglyceride synthesis in rat primary hepatocytes, suggesting that the potential action sites for (-)-ternatin are shared by adipocytes and liver. Although the target molecule of (-)-ternatin remains unknown, our data suggest that (-)-ternatin and its potential target might provide a new therapeutic approach to metabolic disorders. PMID:19463739

  14. Increased NY-ESO-1 expression and reduced infiltrating CD3+ T cells in cutaneous melanoma.

    PubMed

    Giavina-Bianchi, Mara; Giavina-Bianchi, Pedro; Sotto, Mirian Nacagami; Muzikansky, Alona; Kalil, Jorge; Festa-Neto, Cyro; Duncan, Lyn M

    2015-01-01

    NY-ESO-1 is a cancer-testis antigen aberrantly expressed in melanomas, which may serve as a robust and specific target in immunotherapy. NY-ESO-1 antigen expression, tumor features, and the immune profile of tumor infiltrating lymphocytes were assessed in primary cutaneous melanoma. NY-ESO-1 protein was detected in 20% of invasive melanomas (16/79), rarely in in situ melanoma (1/10) and not in benign nevi (0/20). Marked intratumoral heterogeneity of NY-ESO-1 protein expression was observed. NY-ESO-1 expression was associated with increased primary tumor thickness (P = 0.007) and inversely correlated with superficial spreading melanoma (P < 0.02). NY-ESO-1 expression was also associated with reduced numbers and density of CD3+ tumor infiltrating lymphocytes (P = 0.017). When NY-ESO-1 protein was expressed, CD3+ T cells were less diffusely infiltrating the tumor and were more often arranged in small clusters (P = 0.010) or as isolated cells (P = 0.002) than in large clusters of more than five lymphocytes. No correlation of NY-ESO-1 expression with gender, age, tumor site, ulceration, lymph node sentinel status, or survival was observed. NY-ESO-1 expression in melanoma was associated with tumor progression, including increased tumor thickness, and with reduced tumor infiltrating lymphocytes. PMID:25954764

  15. Administration of purified anti-L3T4 monoclonal antibody impairs the resistance of mice to Listeria monocytogenes infection.

    PubMed Central

    Czuprynski, C J; Brown, J F; Young, K M; Cooley, A J

    1989-01-01

    Anti-L3T4 monoclonal antibody (GK1.5) treatment significantly impaired the antilisteria resistance of mice as manifested by increased recovery of listeriae from the spleens and livers of anti-L3T4-treated mice and by greater severity of damage to the liver and other organs. Anti-L3T4-treated mice demonstrated a profound decrease in their T cell-mediated responses to Listeria monocytogenes and its products; they failed to develop delayed type hypersensitivity to soluble listeria antigens in vivo, and their spleen cells proliferated poorly in response to stimulation by either mitogens or listeria antigens in vitro. Spleen cells from control listeria-infected mice produced significant amounts of gamma interferon when stimulated with listeria antigens in vitro, whereas under the same conditions spleen cells from anti-L3T4-treated listeria-infected mice failed to produce detectable gamma interferon. Anti-L3T4 treatment resulted in a slight increase in serum colony-stimulating activity as compared with control listeria-infected mice, probably as a result of the increased bacterial burden in these animals. Despite the dramatic decrease in T-cell activities, anti-L3T4-treated mice succeeded in clearing L. monocytogenes from the spleen and liver in a manner that was only slightly delayed as compared with control listeria-infected mice. In addition, anti-L3T4-treated listeria-immunized mice exhibited a moderate degree of enhanced resistance to rechallenge with L. monocytogenes, and their spleen cells were able to transfer a limited degree of antilisteria resistance to naive recipient mice. Both of these responses, however, were diminished as compared with those of control listeria-immunized mice in the same experiments. Although these observations establish a critical role for L3T4+ cells in the development of maximal resistance to listeriosis, they also suggest that compensatory mechanisms may allow mice to develop considerable antilisteria resistance even when L3T4+ cell

  16. Shp2 suppresses the adipogenic differentiation of preadipocyte 3T3-L1 cells at an early stage

    PubMed Central

    Tao, J; Zheng, L; Meng, M; Li, Y; Lu, Z

    2016-01-01

    Tyrosine phosphatase protein Shp2 is a potential therapeutic target for obesity. However, the mechanism of Shp2 during adipogenesis is not fully understood. The present study investigated the role of Shp2 in the terminal differentiation of preadipocytes. The results showed that Shp2 suppressed adipocyte differentiation in 3T3-L1 cells; overexpression of Shp2 reduced lipid droplet production in 3T3-L1 cells, whereas Shp2 knockdown increased lipid droplet production in 3T3-L1 cells. Furthermore, inhibition of Shp2 activity also enhanced adipocyte differentiation. Interestingly, Shp2 expression was specifically decreased early during differentiation in response to stimulation with the dexamethasone–methylisobutylxanthine–insulin (DMI) hormone cocktail. During the first 2 days of differentiation, Shp2 overexpression impaired the DMI-induced phosphorylation of signal transducer and activator of transcription 3 (STAT3) in 3T3-L1 cells and blocked the peak expression of CCAAT/enhancer-binding proteins β and δ during preadipocyte differentiation. In conclusion, Shp2 downregulated the early stages of hormone-induced differentiation of 3T3-L1 cells and inhibited the expression of the first wave of transcription factors by suppressing the DMI-induced STAT3 signaling pathway. These discoveries point to a novel role of Shp2 during adipogenesis and support the hypothesis that Shp2 could be a therapeutic target for the control of obesity. PMID:27551539

  17. Cellular uptake and fate of fibroin microspheres loaded with randomly fragmented DNA in 3T3 cells

    PubMed Central

    Lee, Jin Sil; Hur, Won

    2016-01-01

    Purified fibroin protein can be obtained in large quantities from silk fibers and processed to form microscopic particles as delivery vehicles for therapeutic agents. In this study, we demonstrated that fibroin microspheres were taken up by 3T3 cells, localized in the nonlysosomal compartment, and secreted from the cytoplasm after medium replenishment. DNA-loaded microspheres were taken up by >95% of 3T3 cells. DNA cargo had no influence on the intracellular trafficking of microspheres, while fluorescently labeled cargo DNA was observed in the lysosomal compartment and in the microspheres. These results indicate that fibroin microspheres can travel through 3T3 cells without making any contact with the lysosomal compartments. The amount of DNA loaded in the microspheres taken up by 3T3 cells was estimated up to 831.0 pg/cell. Thus, fibroin microspheres can deliver a large amount of randomly fragmented DNA (<10 kb) into the cytoplasmic compartment of 3T3 cells. PMID:27257379

  18. STAT5A expression in Swiss 3T3 cells promotes adipogenesis in vivo in an athymic mice model system.

    PubMed

    Stewart, William C; Pearcy, Lisa A; Floyd, Z Elizabeth; Stephens, Jacqueline M

    2011-09-01

    Many studies from our laboratories and others have shown that STAT5 expression and activity are increased during adipogenesis of murine and human adipocytes. Ectopic expression of STAT5A in fibroblasts or preadipocytes can confer or enhance adipogenesis. To determine whether STAT5A also plays a role in adipogenesis in vivo, we injected athymic mice with Swiss 3T3 cells expressing an empty pBABE retrovirus, Swiss cells expressing a pBABE retrovirus-containing STAT5A, or 3T3-F442A preadipocytes. Athymic mice injected with either 3T3-F442A cells or Swiss 3T3 cells expressing STAT5A resulted in fat pad formation at the site of injection. However, mice injected with Swiss cells containing the parent retroviral vector did not have any observable fat pads. An analysis of the ectopic fat pads obtained from the Swiss 3T3 STAT5A mice revealed abundant expression of peroxisome proliferator-activated receptor-γ (PPAR-γ) and adiponectin. The protein levels of both of these fat cell markers were comparable to expression levels in epididymal fat pads. These results demonstrate that STAT5A can promote adipogenesis in vivo in this model system which supports a role of this transcription factor in adipocyte development in the whole animal. PMID:21494231

  19. Mouse osteoblastic cell line (MC3T3-E1) expresses extracellular calcium (Ca2+o)-sensing receptor and its agonists stimulate chemotaxis and proliferation of MC3T3-E1 cells

    NASA Technical Reports Server (NTRS)

    Yamaguchi, T.; Chattopadhyay, N.; Kifor, O.; Butters, R. R. Jr; Sugimoto, T.; Brown, E. M.; O'Malley, B. W. (Principal Investigator)

    1998-01-01

    The calcium-sensing receptor (CaR) is a G protein-coupled receptor that plays key roles in extracellular calcium ion (Ca2+o) homeostasis in parathyroid gland and kidney. Osteoblasts appear at sites of osteoclastic bone resorption during bone remodeling in the "reversal" phase following osteoclastic resorption and preceding bone formation. Bone resorption produces substantial local increases in Ca2+o that could provide a signal for osteoblasts in the vicinity, leading us to determine whether such osteoblasts express the CaR. In this study, we used the mouse osteoblastic, clonal cell line MC3T3-E1. Both immunocytochemistry and Western blot analysis, using an antiserum specific for the CaR, detected CaR protein in MC3T3-E1 cells. We also identified CaR transcripts in MC3T3-E1 cells by Northern analysis using a CaR-specific riboprobe and by reverse transcription-polymerase chain reaction with CaR-specific primers, followed by nucleotide sequencing of the amplified products. Exposure of MC3T3-E1 cells to high Ca2+o (up to 4.8 mM) or the polycationic CaR agonists, neomycin and gadolinium (Gd3+), stimulated both chemotaxis and DNA synthesis in MC3T3-E1 cells. Therefore, taken together, our data strongly suggest that the osteoblastic cell line MC3T3-E1 possesses both CaR protein and mRNA very similar, if not identical, to those in parathyroid and kidney. Furthermore, the CaR in these osteoblasts could play a key role in regulating bone turnover by stimulating the proliferation and migration of such cells to sites of bone resorption as a result of local release of Ca2+o.

  20. A Family-Based Association Study of CYP11A1 and CYP11B1 Gene Polymorphisms With Autism in Chinese Trios.

    PubMed

    Deng, Hong-Zhu; You, Cong; Xing, Yu; Chen, Kai-Yun; Zou, Xiao-Bing

    2016-05-01

    Autism spectrum disorder is a group of neurodevelopmental disorders with the higher prevalence in males. Our previous studies have indicated lower progesterone levels in the children with autism spectrum disorder, suggesting involvement of the cytochrome P-450scc gene (CYP11A1) and cytochrome P-45011beta gene (CYP11B1) as candidate genes in autism spectrum disorder. The aim of this study was to investigate the family-based genetic association between single-nucleotide polymorphisms, rs2279357 in the CYP11A1 gene and rs4534 and rs4541 in the CYP11B1 gene and autism spectrum disorder in Chinese children, which were selected according to the location in the coding region and 5' and 3' regions and minor allele frequencies of greater than 0.05 in the Chinese populations. The transmission disequilibrium test and case-control association analyses were performed in 100 Chinese Han autism spectrum disorder family trios. The genotype and allele frequency of the 3 single-nucleotide polymorphisms had no statistical difference between the children with autism spectrum disorder and their parents (P> .05). Transmission disequilibrium test analysis showed transmission disequilibrium of CYP11A1 gene rs2279357 single-nucleotide polymorphisms (χ(2)= 5.038,P< .001). Our findings provide further support for the hypothesis that a susceptibility gene for autism spectrum disorder exists within or near the CYP11A1 gene in the Han Chinese population. PMID:26690694

  1. Trio study and meta-analysis support the association of genetic variation at the serotonin transporter with early-onset obsessive-compulsive disorder

    PubMed Central

    Walitza, Susanne; Marinova, Zoya; Grünblatt, Edna; Lazic, Stanley E.; Remschmidt, Helmut; Vloet, Timo D.; Wendland, Jens R.

    2014-01-01

    Despite compelling evidence for major genetic contributions to the etiology of obsessive-compulsive disorder (OCD), few genetic variants have been consistently associated with this debilitating illness. Molecular genetic studies in children and adolescents with OCD are of particular interest, since early onset of the disease has been observed to be associated with increased familiality. We replicate here for the first time in early-onset OCD patients, a previously reported association of OCD with the common gain-of-function LA allele at the serotonin transporter linked polymorphic region known as 5-HTTLPR in a collection of parent-offspring trios. The present meta-analysis of this recently refined serotonin transporter gene variant revealed further support for the LA allele conferring increased genetic susceptibility to OCD. We conclude that the 5-HTTLPR is currently the single best supported risk variant for OCD, in regards of early-onset OCD, albeit of modest effect size and the possibility that the conferred risk might not be specific to OCD. PMID:25093702

  2. Influence of the artefact reduction algorithm of Picasso Trio CBCT system on the diagnosis of vertical root fractures in teeth with metal posts

    PubMed Central

    Bezerra, I S Q; Neves, F S; Vasconcelos, T V; Ambrosano, G M B

    2015-01-01

    Objectives: To assess the influence of the artefact reduction algorithm (AR) available on the Picasso Trio 3D® imaging system (Vatech, Hwaseong, Republic of Korea) on image quality [greyscale values, contrast-to-noise ratio (CNR) and artefact formation] and diagnosis of vertical root fractures (VRFs) in the teeth with intracanal metal posts. Methods: 30 uniradicular teeth had their crowns removed and their roots endodontically treated to receive intracanal metal posts. In 20 teeth, both complete (n = 10) and incomplete (n = 10) VRFs were created. Each tooth was scanned twice, with and without AR activation. The mean and variation of greyscale values, as well as CNR, were calculated for all images. Subsequently, an evaluator compared the amount of artefact (cupping, white streaks and dark bands) in all images. Five evaluators rated for VRF presence using a five-point scale. Results: Mean greyscale values and CNR were significantly decreased in images acquired with the AR. The usage of the algorithm promoted an overall reduction of image artefacts. Regarding the diagnosis of complete and incomplete VRFs, the use of the AR had an overall negative impact on specificity and accuracy. Conclusions: While indeed reducing artefact formation, the use of the AR, instead of improving the impact on the diagnosis of VRFs in teeth with intracanal metal posts, had a negative impact on the diagnosis. PMID:25764360

  3. The UNC-73/Trio RhoGEF-2 domain is required in separate isoforms for the regulation of pharynx pumping and normal neurotransmission in C. elegans.

    PubMed

    Steven, Robert; Zhang, Lijia; Culotti, Joseph; Pawson, Tony

    2005-09-01

    In both Caenorhabditis elegans and Drosophila, UNC-73/Trio functions in axon guidance by signaling through the Rac GTPase to regulate cytoskeletal rearrangements necessary for growth cone migrations. Here, we show that the complex C. elegans unc-73 gene encodes at least eight differentially expressed UNC-73 intracellular protein isoforms. Previously reported mutations affecting UNC-73 isoforms encoding the Rac-specific RhoGEF-1 domain cause uncoordinated movement, correlating with defects in axon guidance. Mutations in isoforms encoding the Rho-specific RhoGEF-2 domain, which we describe here, result in L1 stage larval lethality with no associated axon guidance defects. Isoform-specific rescue experiments reveal separate functions for the various RhoGEF-2-containing UNC-73 isoforms, which would not likely be discovered by conventional genetic screening. UNC-73 D1 and D2 appear to function redundantly in pharynx muscle to regulate the rate and strength of pharynx pumping, and in the HSN neurons and vulval muscles to control egg laying. Isoforms C1, C2, E, and F act redundantly within the nervous system to regulate the speed of locomotion. The multiple UNC-73 isoforms containing Rac- and Rho-specific RhoGEF domains therefore have distinct physiological functions. In addition to its previously identified role involving RhoGEF-1 in migrating cells and growth cones, our data indicate that UNC-73 signals through RhoGEF-2 to regulate pharynx and vulva musculature and to modulate synaptic neurotransmission. PMID:16140983

  4. CREB Activation Induces Adipogenesis in 3T3-L1 Cells

    PubMed Central

    Reusch, Jane E. B.; Colton, Lilliester A.; Klemm, Dwight J.

    2000-01-01

    Obesity is the result of numerous, interacting behavioral, physiological, and biochemical factors. One increasingly important factor is the generation of additional fat cells, or adipocytes, in response to excess feeding and/or large increases in body fat composition. The generation of new adipocytes is controlled by several “adipocyte-specific” transcription factors that regulate preadipocyte proliferation and adipogenesis. Generally these adipocyte-specific factors are expressed only following the induction of adipogenesis. The transcription factor(s) that are involved in initiating adipocyte differentiation have not been identified. Here we demonstrate that the transcription factor, CREB, is constitutively expressed in preadipocytes and throughout the differentiation process and that CREB is stimulated by conventional differentiation-inducing agents such as insulin, dexamethasone, and dibutyryl cAMP. Stably transfected 3T3-L1 preadipocytes were generated in which we could induce the expression of either a constitutively active CREB (VP16-CREB) or a dominant-negative CREB (KCREB). Inducible expression of VP16-CREB alone was sufficient to initiate adipogenesis as determined by triacylglycerol storage, cell morphology, and the expression of two adipocyte marker genes, peroxisome proliferator activated receptor gamma 2, and fatty acid binding protein. Alternatively, KCREB alone blocked adipogenesis in cells treated with conventional differentiation-inducing agents. These data indicate that activation of CREB was necessary and sufficient to induce adipogenesis. Finally, CREB was shown to bind to putative CRE sequences in the promoters of several adipocyte-specific genes. These data firmly establish CREB as a primary regulator of adipogenesis and suggest that CREB may play similar roles in other cells and tissues. PMID:10629058

  5. Analysis of Suppressor and Non-Suppressor FOXP3+ T Cells in HIV-1-Infected Patients

    PubMed Central

    Arruvito, Lourdes; Baz, Plácida; Billordo, Luis A.; Lasala, Maria B.; Salomón, Horacio; Geffner, Jorge; Fainboim, Leonardo

    2012-01-01

    Recently, it was shown that peripheral blood FOXP3+CD4+ T cells are composed of three phenotypic and functionally distinct subpopulations. Two of them having in vitro suppressive effects were characterized as resting Treg cells (rTregs) and activated Treg cells (aTregs). A third subset, identified as FOXP3+ non-Tregs, does not display any suppressor activity and produce high levels of Th1 and Th17 cytokines upon stimulation. In the present study we focus on the characteristics of these three subsets of FOXP3+CD4+ T cells in untreated HIV-1-infected patients. We found that the absolute counts of rTregs, aTregs and FOXP3+ non-Tregs were reduced in HIV-1 patients compared with healthy donors. The relative frequency of rTregs and aTregs was similar in HIV-1 patients and healthy donors, while the frequency of FOXP3+ non-Tregs was significantly higher in HIV-1 patients, reaching a maximum in those patients with the lower values of CD4 counts. Contrasting with the observations made in FOXP3- CD4+ T cells, we did not find a negative correlation between the number of rTregs, aTregs or FOXP3+ non-Tregs and virus load. Studies performed with either whole PBMCs or sorted aTregs and FOXP3+ non-Tregs cells showed that these two populations of FOXP3+ T cells were highly permissive to HIV-1 infection. Upon infection, FOXP3+ non-Tregs markedly down-regulates its capacity to produce Th1 and Th17 cytokines, however, they retain the ability to produce substantial amounts of Th2 cytokines. This suggests that FOXP3+ non-Tregs might contribute to the polarization of CD4+ T cells into a Th2 profile, predictive of a poor outcome of HIV-1-infected patients. PMID:23285102

  6. Normal-appearing brain tissue analysis in radiologically isolated syndrome using 3 T MRI.

    PubMed

    Labiano-Fontcuberta, Andrés; Mato-Abad, Virginia; Álvarez-Linera, Juan; Hernández-Tamames, Juan Antonio; Martínez-Ginés, María Luisa; Aladro, Yolanda; Ayuso, Lucía; Domingo-Santos, Ángela; Benito-León, Julián

    2016-07-01

    To date, it remains largely unknown whether there is in radiologically isolated syndrome (RIS) brain damage beyond visible T2 white matter lesions. We used single- voxel proton magnetic resonance spectroscopy and diffusion tensor imaging (3 T MRI) to analyze normal-appearing brain tissue regions in 18 RIS patients and 18 matched healthy controls. T2-hyperintense lesion volumes and structural brain volumes were also measured. The absolute metabolite concentrations and ratios of total N-acetylaspartate+N-acetylaspartyl glutamate (NAA), choline-containing compounds, myoinositol, and glutamine-glutamate complex to creatine were calculated. Spectral analysis was performed by LCModel. Voxelwise morphometry analysis was performed to localize regions of brain tissue showing significant changes of fractional anisotropy or mean diffusivity. Compared with healthy controls, RIS patients did not show any significant differences in either the absolute concentration of NAA or NAA/Cr ratio in mid-parietal gray matter. A trend toward lower NAA concentrations (-3.35%) was observed among RIS patients with high risk for conversion to multiple sclerosis. No differences in the other metabolites or their ratios were observed. RIS patients showed lower fractional anisotropy only in clusters overlapping lesional areas, namely in the cingulate gyrus bilaterally and the frontal lobe subgyral bilaterally (P < 0.001). Normalized brain and cortical volumes were significantly lower in RIS patients than in controls (P = 0.01 and P = 0.03, respectively). Our results suggest that in RIS, global brain and cortical atrophy are not primarily driven by significant occult microstructural normal appearing brain damage. Longitudinal MRI studies are needed to better understand the pathological processes underlying this novel entity. PMID:27399108

  7. Effects of lipoic acid on lipolysis in 3T3-L1 adipocytes[S

    PubMed Central

    Fernández-Galilea, Marta; Pérez-Matute, Patricia; Prieto-Hontoria, Pedro L; Martinez, J Alfredo; Moreno-Aliaga, Maria J

    2012-01-01

    Lipoic acid (LA) is a naturally occurring compound with beneficial effects on obesity. The aim of this study was to evaluate its effects on lipolysis in 3T3-L1 adipocytes and the mechanisms involved. Our results revealed that LA induced a dose- and time-dependent lipolytic action, which was reversed by pretreatment with the c-Jun N-terminal kinase inhibitor SP600125, the PKA inhibitor H89, and the AMP-activated protein kinase activator AICAR. In contrast, the PI3K/Akt inhibitor LY294002 and the PDE3B antagonist cilostamide enhanced LA-induced lipolysis. LA treatment for 1 h did not modify total protein content of hormone-sensitive lipase (HSL) but significantly increased the phosphorylation of HSL at Ser563 and at Ser660, which was reversed by H89. LA treatment also induced a marked increase in PKA-mediated perilipin phosphorylation. LA did not significantly modify the protein levels of adipose triglyceride lipase or its activator comparative gene identification 58 (CGI-58) and inhibitor G(0)/G(1) switch gene 2 (G0S2). Furthermore, LA caused a significant inhibition of adipose-specific phospholipase A2 (AdPLA) protein and mRNA levels in parallel with a decrease in the amount of prostaglandin E2 released and an increase in cAMP content. Together, these data suggest that the lipolytic actions of LA are mainly mediated by phosphorylation of HSL through cAMP-mediated activation of protein kinase A probably through the inhibition of AdPLA and prostaglandin E2. PMID:22941773

  8. Lithium differentially affects clock gene expression in serum-shocked NIH-3T3 cells.

    PubMed

    Osland, Teresa M; Fernø, Johan; Håvik, Bjarte; Heuch, Ivar; Ruoff, Peter; Lærum, Ole Didrik; Steen, Vidar M

    2011-07-01

    Bipolar disorder has been associated with disturbances in circadian rhythms. Lithium is frequently used in the long-term treatment of bipolar disorder, and has been shown to prolong such rhythms in animals and humans. To examine whether lithium affects the expression of genes regulating the circadian clock, cultured NIH-3T3 cells were synchronized by serum-shocking, and the relative expression of the clock genes Period1 (Per1), Period2 (Per2), Period3 (Per3), Cryptochrome1 (Cry1), Cryptochrome2 (Cry2), Brain and muscle aryl hydrocarbon nuclear translocator-like 1 (Bmal1), Circadian locomotor output cycles kaput (Clock), Rev-Erb-α (Nr1d1), RAR-related orphan receptor α (Ror-α), Glycogen synthase kinase-3β (Gsk-3β), Casein kinase 1-ε (CK1-ε; Csnk1ε), E4 binding protein 4 (E4BP4; Nfil-3) and albumin D-binding protein (Dbp) was examined for three consecutive days in the presence of lithium (20 mM) or vehicle (20 mM NaCl). We found that lithium significantly increased the expression of Per2 and Cry1, whereas Per3, Cry2, Bmal1, E4BP4 and Rev-Erb-α expression was reduced. We also found that lithium prolonged the period of Per2. Taken together, these effects on clock gene expression may be relevant for the effects of lithium on biological rhythms and could also give new leads to further explore its mood-stabilizing actions in the treatment of bipolar disorder. PMID:20837565

  9. 3T MR Spectroscopy Reveals an Imbalance between Excitatory and Inhibitory Neurotransmitters in Amyotrophic Lateral Sclerosis

    PubMed Central

    Foerster, Bradley R.; Pomper, Martin G.; Callaghan, Brian C.; Petrou, Myria; Edden, Richard A.E.; Mohamed, Mona A.; Welsh, Robert C.; Carlos, Ruth C.; Barker, Peter B.; Feldman, Eva L.

    2015-01-01

    Objective To determine whether there are reductions in γ-aminobutyric acid (GABA) and elevations of glutamate + glutamine (Glx) levels in different brain regions of patients with amyotrophic lateral sclerosis (ALS) using proton magnetic resonance spectroscopy (1H-MRS). Design 3T short echo time and GABA-edited 1H-MRS centered on the left motor cortex and left subcortical white matter. Short echo time 1H-MRS was also performed centered on the pons. Data were analyzed using logistic regression, t-tests, and Pearson correlations. Post hoc analyses were performed to investigate differences between riluzole-naïve and riluzole-treated ALS patients. Participants Twenty-nine ALS patients and thirty age- and gender-matched healthy controls (HCs). Results ALS patients had significantly lower levels of GABA in the motor cortex compared to HCs (P<.01). ALS patients also had significantly lower levels of N-acetylaspartate in the motor cortex (P<.01), subcortical white matter (P<.05), and pons (P<.01) and higher levels of myo-inositol in the motor cortex (P<.001) and subcortical white matter (P<.01) compared to HCs. Compared to riluzole-treated ALS patients, riluzole-naïve ALS patients had higher levels of Glx in the motor cortex (P<.05) and pons (P<.01), higher levels of creatine in the motor cortex (P<.001) and subcortical white matter (P=.05), and higher levels of N-acetylaspartate in the motor cortex (P<.01). Conclusion There are reduced levels of GABA in the motor cortex of ALS patients. There are elevations of Glx in riluzole-naïve ALS patients compared to ALS riluzole-treated patients. These results point to an imbalance between excitatory and inhibitory neurotransmission, contributing to the pathogenesis of ALS. PMID:23797905

  10. Compartmentalized Ras Proteins Transform NIH 3T3 Cells with Different Efficiencies▿ †

    PubMed Central

    Cheng, Chiang-Min; Li, Huiling; Gasman, Stéphane; Huang, Jian; Schiff, Rachel; Chang, Eric C.

    2011-01-01

    Ras GTPases were long thought to function exclusively from the plasma membrane (PM). However, a current model suggests that Ras proteins can compartmentalize to regulate different functions, and an oncogenic H-Ras mutant that is restricted to the endomembrane can still transform cells. In this study, we demonstrated that cells transformed by endomembrane-restricted oncogenic H-Ras formed tumors in nude mice. To define downstream targets of endomembrane Ras pathways, we analyzed Cdc42, which concentrates in the endomembrane and has been shown to act downstream of Ras in Schizosaccharomyces pombe. Our data show that cell transformation induced by endomembrane-restricted oncogenic H-Ras was blocked when Cdc42 activity was inhibited. Moreover, H-Ras formed a complex with Cdc42 on the endomembrane, and this interaction was enhanced when H-Ras was GTP bound or when cells were stimulated by growth factors. H-Ras binding evidently induced Cdc42 activation by recruiting and/or activating Cdc42 exchange factors. In contrast, when constitutively active H-Ras was restricted to the PM by fusing to a PM localization signal from the Rit GTPase, the resulting protein did not detectably activate Cdc42 although it activated Raf-1 and efficiently induced hallmarks of Ras-induced senescence in human BJ foreskin fibroblasts. Surprisingly, PM-restricted oncogenic Ras when expressed alone could only weakly transform NIH 3T3 cells; however, when constitutively active Cdc42 was coexpressed, together they transformed cells much more efficiently than either one alone. These data suggest that efficient cell transformation requires Ras proteins to interact with Cdc42 on the endomembrane and that in order for a given Ras protein to fully transform cells, multiple compartment-specific Ras pathways need to work cooperatively. PMID:21189290

  11. Triiodothyronine modulates the expression of leptin and adiponectin in 3T3-L1 adipocytes

    PubMed Central

    de Oliveira, Miriane; Síbio, Maria Teresa De; Olimpio, Regiane Marques Castro; Moretto, Fernanda Cristina Fontes; Luvizotto, Renata de Azevedo Melo; Nogueira, Celia Regina

    2015-01-01

    Objective To study the effect of different doses of triiodothyronine on gene expression of the adipokines leptin and adiponectin, at different times, and to evaluate the difference in expression between the two adipokines in each group. Methods 3T3-L1 adipocytes were incubated with triiodothyronine at physiological dose (10nM) and supraphysiological doses (100nM or 1,000nM), or without triiodothyronine (control, C) for 0.5, 6, or 24 hours. Leptin and adiponectin mRNA was detected using real-time polymerase chain reaction (RT-PCR). One-way analyses of variance, Tukey’s test or Student’s t test, were used to analyze data, and significance level was set at 5%. Results Leptin levels decreased in the 1,000nM-dose group after 0.5 hour. Adiponectin levels dropped in the 10nM-dose group, but increased at the 100nM dose. After 6 hours, both genes were suppressed in all hormone concentrations. After 24 hours, leptin levels increased at 10, 100 and 1,000nM groups as compared to the control group; and adiponectin levels increased only in the 100nM group as compared to the control group. Conclusion These results demonstrated fast actions of triiodothyronine on the leptin and adiponectin expression, starting at 0.5 hour, at a dose of 1,000nM for leptin and 100nM for adiponectin. Triiodothyronine stimulated or inhibited the expression of adipokines in adipocytes at different times and doses which may be useful to assist in the treatment of obesity, assuming that leptin is increased and adiponectin is decreased, in obesity cases. PMID:25993072

  12. Crystal structure and chemistry of lithium-bearing trioctahedral micas-3T

    USGS Publications Warehouse

    Brigatti, M.F.; Kile, D.E.; Poppi, L.

    2003-01-01

    Chemical analyses and crystal structure refinements were performed on lithian siderophyllite-3T crystals from granitic pegmatites of the anorogenic Pikes Peak batholith (Colorado) to characterize the crystal chemistry and relations with trioctahedral lithium-bearing micas showing different stacking sequences. Chemical data show that the studied samples fall on the siderophyllite-polylithionite join, closer to the siderophyllite end-member. Single-crystal X-ray refinements were carried out on three samples (two of which were taken from core and rim of the same crystal) in space-group P31 12 (the agreement factor, Robs, varies between 0.034 and 0.036). Mean bond distances and mean electron counts of M1, M2 and M3 octahedral sites indicate an ordered cation distribution with M1 and M3 positions substantially larger than M2. In the sample with the largest iron content, the M2 mean electron count increases as well as the mean distance, whereas remains smaller than or . The tetrahedral cation-oxygen atom mean distances range from 1.614 to 1.638 A?? and from 1.663 to 1.678 A?? for T1 and T2 sites, respectively, being consistent with Al3+ enrichment in the T2 sites. The tetrahedral rotation angle, ??, is generally small (3.1 ??? ?? ??? 4.6??) and decreases with siderophyllite content. As Fe increases, the T1 tetrahedron becomes flatter (112.4 ??? t??1 ??? 110.5??), whereas T2 tetrahedron distortion appears unchanged (110.7 ??? t??T2 ??? 110.9).

  13. Exogenous Sodium Pyruvate Stimulates Adipogenesis of 3T3-L1 Cells.

    PubMed

    Hwang, Ji-Sun; Kim, Song-Yi; Jung, Eun-Hye; Kwon, Mi-Youn; Kim, Kyoung-Hong; Cho, Hyeongjin; Han, Inn-Oc

    2016-01-01

    We investigated the effects of exogenous sodium pyruvate (SP) on adipocyte differentiation, lipid accumulation, and the mRNA expression levels of adipogenesis-related genes in 3T3-L1 pre-adipocytes. Differentiation of pre-adipocytes was induced by MDI (3-isobutyl-1-methylxanthine: IBMX, dexamethasone: DEX, and insulin), in the presence or absence of SP. Adipogenesis was stimulated by SP in a concentration-dependent manner. SP also induced the expression of genes encoding aP2, GLUT4, and adiponectin, but had no effect on cell proliferation. Exogenous glucose did not promote adipogenesis or lipid accumulation. 2-deoxy-D-glucose inhibited adipogenesis initiated by MDI, but failed to influence the effects of SP on adipogenesis, whereas 3-bromopyruvate inhibited adipogenesis regardless of whether SP was present. The pro-adipogenic properties of SP were limited to the early events of adipogenesis. To determine whether SP mimics the adipogenic action of dexamethasone or insulin, we examined the effects of SP on adipogenesis with combinations of IBMX, DEX, and insulin. SP did not improve incomplete lipid accumulation observed in cells grown under IBMX-, DEX-, or insulin-free conditions. Insulin-stimulated ERK1/2 phosphorylation was diminished by SP, while phosphorylation of Akt was increased, correlating with increased glucose uptake in response to insulin. We also observed that SP stimulated immediate early expression of C/EBPβ and C/EBPδ. The PPARγ antagonist GW9662 inhibited adipogenesis. Our findings highlight the adipogenic function of exogenous SP by stimulating early events of adipogenesis. PMID:26053972

  14. A surface-tethered spheroid model for functional evaluation of 3T3-L1 adipocytes.

    PubMed

    Turner, Paul A; Harris, Lacey M; Purser, Christine A; Baker, Rodney C; Janorkar, Amol V

    2014-01-01

    In order to effectively treat obesity, it must be better understood at the cellular level with respect to metabolic state and environmental stress. However, current two-dimensional (2D) in vitro cell culture methods do not represent the in vivo adipose tissue appropriately due to the absence of complex architecture and cellular signaling. Conversely, 3D in vitro cultures have been reported to have optimal results mimicking the adipose tissue in vivo. The main aim of this study was to examine the efficacy of a novel conjugate of a genetically engineered polymer, elastin-like polypeptide (ELP) and a synthetic polymer, polyethyleneimine (PEI), toward creating a 3D preadipocyte culture system. We then used this 3D culture model to study the preadipocyte differentiation and adipocyte maintenance processes when subjected to various dosages of nutritionally relevant free fatty acids with respect to total DNA and protein content, cell viability, and intracellular triglyceride accumulation. Our results showed that 3T3-L1 preadipocytes cultured on the ELP-PEI surface formed 3D spheroids within 72 h, whereas the cells cultured on unmodified tissue culture polystyrene surfaces remained in monolayer configuration. Significant statistical differences were discovered between the 3D spheroid and 2D monolayer culture with respect to the DNA and protein content, fatty acid consumption, and triglyceride accumulation, indicating differences in cellular response. Results indicated that the 3D culture may be a more sensitive modeling technique for in vitro adipocyte culture and provides a platform for future evaluation of 3D in vitro adipocyte function. PMID:24038000

  15. Mitogenic stimuli and phosphatidylinositol (PI) turnover in cultured 3T3 fibroblasts

    SciTech Connect

    Kohler, C.; Petersen, R.

    1986-03-01

    The hydrolysis of PI and polyphosphoinositides by phopholipase C is an early and rapid response to cell activation by a variety of neurotransmitters, hormones, growth factors and pharmacological agonists. The authors have examined the role of PI turnover and the generation of second messengers (diacylglycerol and inositol trisphosphate) in the mitogenic response of cultured Balb/c and Swiss 3T3 cells to polypeptide growth factors. Cells were prelabelled with /sup 3/H inositol for 18-20 hours, washed and suspended in Herpes + Li/sup +/ buffer, and stimulated with platelet-derived growth factor (PDGF), vasopressin, insulin, and other growth factors. PI turnover was measured as the increase in total inositol phosphate (IP) production. IP1, IP2, and IP3 were characterized by sequential elution from a Dowex column. Partially purified PDGF produced a 2-4 fold stimulation of total IP production. This was seen as early as 30 seconds after stimulation and increased for up to 1-2 hours. Balb/c cells were more sensitive than Swiss cells to the mitogenic and PI effects of PDGF. Other mitogenic stimuli had differential effects on PI turnover. Vasopressin (4-400 ng/ml) markedly stimulated PI turnover (3-6 fold) in Swiss, but not Balb/c cells. Insulin (100 ng/ml - 10 ..mu..g/ml) increased total IP to a greater degree in Balb/c cells. Epidermal growth factor (10 ng/ml - 10 ..mu..g/ml) had no effect on PI turnover and fibroblast growth factor (10 ng/ml - 10 ..mu..g/ml) only stimulated at the higher concentrations in Swiss cells. Thrombin (1U/ml - 10 U/ml) produced a 1.5 - 2 fold stimulation in Balb/c cells. Thus, various polypeptide growth factors have differential effects on PI turnover depending on their mitogenic potential and the effector cell type.

  16. High-Resolution 3 T MR Microscopy Imaging of Arterial Walls

    SciTech Connect

    Sailer, Johannes Rand, Thomas; Berg, Andreas; Sulzbacher, Irene; Peloschek, P.; Hoelzenbein, Thomas; Lammer, Johannes

    2006-10-15

    Purpose. To achieve a high spatial resolution in MR imaging that allows for clear visualization of anatomy and even histology and documentation of plaque morphology in in vitro samples from patients with advanced atherosclerosis. A further objective of our study was to evaluate whether T2-weighted high-resolution MR imaging can provide accurate classification of atherosclerotic plaque according to a modified American Heart Association classification. Methods. T2-weighted images of arteries were obtained in 13 in vitro specimens using a 3 T MR unit (Medspec 300 Avance/Bruker, Ettlingen, Germany) combined with a dedicated MR microscopy system. Measurement parameters were: T2-weighted sequences with TR 3.5 sec, TE 15-120 msec; field of view (FOV) 1.4 x 1.4; NEX 8; matrix 192; and slice thickness 600 {mu}m. MR measurements were compared with corresponding histologic sections. Results. We achieved excellent spatial and contrast resolution in all specimens. We found high agreement between MR images and histology with regard to the morphology and extent of intimal proliferations in all but 2 specimens. We could differentiate fibrous caps and calcifications from lipid plaque components based on differences in signal intensity in order to differentiate hard and soft atheromatous plaques. Hard plaques with predominantly intimal calcifications were found in 7 specimens, and soft plaques with a cholesterol/lipid content in 5 cases. In all specimens, hemorrhage or thrombus formation, and fibrotic and hyalinized tissue could be detected on both MR imaging and histopathology. Conclusion. High-resolution, high-field MR imaging of arterial walls demonstrates the morphologic features, volume, and extent of intimal proliferations with high spatial and contrast resolution in in vitro specimens and can differentiate hard and soft plaques.

  17. Trichinella spiralis infection suppressed gut inflammation with CD4(+)CD25(+)Foxp3(+) T cell recruitment.

    PubMed

    Cho, Min Kyoung; Park, Mi Kyung; Kang, Shin Ae; Choi, Seon Hee; Ahn, Soon Cheol; Yu, Hak Sun

    2012-12-01

    In order to know the effect of pre-existing Trichinella spiralis infection on experimentally induced intestinal inflammation and immune responses, we induced colitis in T. spiralis-infected mice and observed the severity of colitis and the levels of Th1, Th2, and regulatory cytokines and recruitment of CD4(+)CD25(+)Foxp3(+) T (regulatory T; T(reg)) cells. Female C57BL/6 mice were infected with 250 muscle larvae; after 4 weeks, induction of experimental colitis was performed using 3% dextran sulfate sodium (DSS). During the induction period, we observed severity of colitis, including weight loss and status of stool, and evaluated the disease activity index (DAI). A significantly low DAI and degree of weight loss were observed in infected mice, compared with uninfected mice. In addition, colon length in infected mice was not contracted, compared with uninfected mice. We also observed a significant increase in production of pro-inflammatory cytokines, IL-6 and IFN-γ, in spleen lymphocytes treated with DSS; however, such an increase was not observed in infected mice treated with DSS. Of particular interest, production of regulatory cytokines, IL-10 and transforming growth factor (TGF)-β, in spleen lymphocytes showed a significant increase in mice infected with T. spiralis. A similar result was observed in mesenteric lymph nodes (MLN). Subsets of the population of T(reg) cells in MLN and spleen showed significant increases in mice infected with T. spiralis. In conclusion, T. spiralis infection can inhibit the DSS-induced colitis in mice by enhancing the regulatory cytokine and T(reg) cells recruitment. PMID:23230342

  18. A Partially Purified Acinetobacter baumannii Phage Preparation Exhibits no Cytotoxicity in 3T3 Mouse Fibroblast Cells

    PubMed Central

    Henein, Alexandra E.; Hanlon, Geoffrey W.; Cooper, Callum J.; Denyer, Stephen P.; Maillard, Jean-Yves

    2016-01-01

    A surge in the level and scale of antibiotic resistance has prompted renewed interest in the application of bacteriophages to treat bacterial infections. However, concerns still exist over their efficacy and safety. Acinetobacter baumannii phage BS46, a member of the family Myoviridae, has previously been shown to be effective in murine models. The cytotoxic effect of this phage was evaluated in mouse fibroblast 3T3 cells using four different assays: trypan blue; staining with Hoechst and propidium iodide; lactate dehydrogenase release; and the MTS assay. The addition of phage concentrations up to 2 × 109 pfu/mL showed little to no impact on the viability of 3T3 cells after 24 h exposure using the different assays. This study demonstrates that phage BS46 is non-cytotoxic to 3T3 cells using four different assays and that appropriate quality assurance protocols for phage therapeutics are required. PMID:27536286

  19. Prolonged treatment with 3-isobutyl-1-methylxanthine improves the efficiency of differentiating 3T3-L1 cells into adipocytes.

    PubMed

    Hua, Yongjie; Ke, Shanshan; Wang, Yao; Irwin, David M; Zhang, Shuyi; Wang, Zhe

    2016-08-15

    Until now, the low efficiency of current protocols or kits for the differentiation of 3T3-L1 preadipocytes makes it difficult to continue the studies of the cellular and molecular mechanisms in adipocytes. Here we present a productive and highly efficient protocol for the differentiation of 3T3-L1 cells that uses a prolonged treatment with 3-isobutyl-1-methylxanthine (IBMX) during the differentiated process. 3T3-L1 cells of unknown passage +3 and unknown passage +7 treated with a prolonged exposure to IBMX showed significantly increased differentiation efficiency by day 15, in contrast to low levels of differentiation seen with protocols that lacked additional IBMX. PMID:27210514

  20. A Partially Purified Acinetobacter baumannii Phage Preparation Exhibits no Cytotoxicity in 3T3 Mouse Fibroblast Cells.

    PubMed

    Henein, Alexandra E; Hanlon, Geoffrey W; Cooper, Callum J; Denyer, Stephen P; Maillard, Jean-Yves

    2016-01-01

    A surge in the level and scale of antibiotic resistance has prompted renewed interest in the application of bacteriophages to treat bacterial infections. However, concerns still exist over their efficacy and safety. Acinetobacter baumannii phage BS46, a member of the family Myoviridae, has previously been shown to be effective in murine models. The cytotoxic effect of this phage was evaluated in mouse fibroblast 3T3 cells using four different assays: trypan blue; staining with Hoechst and propidium iodide; lactate dehydrogenase release; and the MTS assay. The addition of phage concentrations up to 2 × 10(9) pfu/mL showed little to no impact on the viability of 3T3 cells after 24 h exposure using the different assays. This study demonstrates that phage BS46 is non-cytotoxic to 3T3 cells using four different assays and that appropriate quality assurance protocols for phage therapeutics are required. PMID:27536286

  1. 26 CFR 1.904(g)-3T - Ordering rules for the allocation of net operating losses, net capital losses, U.S. source losses...

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ...(g)-3T Section 1.904(g)-3T Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY (CONTINUED) INCOME TAX (CONTINUED) INCOME TAXES Income from Sources Without the United States § 1.904(g)-3T.... The rules must be applied in the order set forth in paragraphs (b) through (g) of this section....

  2. 26 CFR 1.904(g)-3T - Ordering rules for the allocation of net operating losses, net capital losses, U.S. source losses...

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ...(g)-3T Section 1.904(g)-3T Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY....904(g)-3T Ordering rules for the allocation of net operating losses, net capital losses, U.S. source... domestic losses. The rules must be applied in the order set forth in paragraphs (b) through (g) of...

  3. 26 CFR 1.904(g)-3T - Ordering rules for the allocation of net operating losses, net capital losses, U.S. source losses...

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ...(g)-3T Section 1.904(g)-3T Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY....904(g)-3T Ordering rules for the allocation of net operating losses, net capital losses, U.S. source... domestic losses. The rules must be applied in the order set forth in paragraphs (b) through (g) of...

  4. Skin tumor responsiveness to interleukin-2 treatment and CD8 Foxp3+ T cell expansion in an immunocompetent mouse model.

    PubMed

    Foureau, David M; McKillop, Iain H; Jones, Chase P; Amin, Asim; White, Richard L; Salo, Jonathan C

    2011-09-01

    Recombinant human interleukin-2 (rhIL-2) therapy is approved for treating patients with advanced melanoma yet significant responses are observed in only 10-15% of patients. Interleukin-2 induces Foxp3 expression in activated human CD8 T cells in vitro and expands circulating CD8 Foxp3+ T cells in melanoma patients. Employing IL-2 responsive (B16-F1, B16-BL6, JB/MS, MCA-205) and nonresponsive (JB/RH, B16-F10) subcutaneous tumor mouse models, we evaluated CD8 Foxp3+ T cell distribution and changes in response to rhIL-2 (50,000 U, i.p. or s.q., twice daily for 5 days). In tumor-free mice and subcutaneous tumor-bearing mouse models, CD8 Foxp3+ T cells were a rare but naturally occurring cell subset. Primarily located in skin-draining lymph nodes, CD8 Foxp3+ T cells expressed both activated T cell (CD28(+), CD44(+)) and Treg (CTLA4(+), PD1(lo/var), NKG2A(+/var)) markers. Following treatment with rhIL-2, a dramatic increase in CD8 Foxp3+ T cell prevalence was observed in the circulation and tumor-draining lymph nodes (TD.LNs) of animals bearing IL-2 nonresponsive tumors, while no significant changes were observed in the circulation and TD.LNs of animals bearing IL-2 responsive tumors. These findings suggest expansion of CD8 Foxp3+ T cell population in response to rhIL-2 treatment may serve as an early marker for tumor responsiveness to immunotherapy in an immune competent model. Additionally, these data may provide insight to predict response in patients with melanoma undergoing rhIL-2 treatment. PMID:21638127

  5. High glucose induces autophagy of MC3T3-E1 cells via ROS-AKT-mTOR axis.

    PubMed

    Wang, Xiaoju; Feng, Zhengping; Li, Jiling; Chen, Lixue; Tang, Weixue

    2016-07-01

    In the present study, we investigate the function of ROS-AKT-mTOR axis on the apoptosis, proliferation and autophagy of MC3T3-E1 cells, and the proliferation of MC3T3-E1 cells after autophagy inhibition under high glucose conditions. MC3T3-E1 cells cultured in vitro were divided into the following groups: normal control group, N-acetylcysteine (NAC) group, 11.0 mM high glucose group, 11.0 mM high glucose + NAC group, 22.0 mM high glucose group, 22.0 mM high glucose + NAC group, CQ group, 22.0 mM high glucose + CQ group, 3-MA group and 3-MA + 22.0 mM high glucose group. ROS production was measured by DCFH-DA fluorescent probe. Cell proliferation was measured by MTT assay. Cells in different groups were stained with Annexin V-FITC/PI, and then apoptosis rate was detected by flow cytometry. Nucleus morphology was observed under fluorescence microscope after being incubated with Honchest33258. Protein expression was measured using Western blotting and immunofluorescence. Cell apoptosis and proliferation in high glucose group were increased and decreased, respectively, in a dose-dependent manner. Autophagy was significantly induced in high glucose group, even though different concentration of glucose induced autophagy in different stages of autophagy. ROS production in MC3T3-E1 cells was remarkably increased in high glucose group, but not in a dose-dependent manner. NAC, as an antioxidant, reduced ROS production and ameliorated cell apoptosis, proliferation abnormity and autophagy caused by high glucose. Expression of p-AKT and p-mTOR proteins were dramatically decreased in high glucose group, and NAC reversed their expression. In addition, 3-MA, an inhibitor of autophagy, significantly decreased the proliferation of MC3T3-E1 cells. When cocultured with 22.0 mM glucose that induced autophagy, proliferation of MC3T3-E1 cells was not affected compared to 22.0 mM high glucose group. Our present findings reveal that high glucose affects apoptosis

  6. Parasitic Nematode-Induced CD4+Foxp3+T Cells Can Ameliorate Allergic Airway Inflammation

    PubMed Central

    Kang, Shin Ae; Park, Mi-Kyung; Cho, Min Kyoung; Park, Sang Kyun; Jang, Min Seong; Yang, Bo-Gie; Jang, Myoung Ho; Kim, Dong-Hee; Yu, Hak Sun

    2014-01-01

    Background The recruitment of CD4+CD25+Foxp3+T (Treg) cells is one of the most important mechanisms by which parasites down-regulate the immune system. Methodology/Principal Findings We compared the effects of Treg cells from Trichinella spiralis-infected mice and uninfected mice on experimental allergic airway inflammation in order to understand the functions of parasite-induced Treg cells. After four weeks of T. spiralis infection, we isolated Foxp3-GFP-expressing cells from transgenic mice using a cell sorter. We injected CD4+Foxp3+ cells from T. spiralis-infected [Inf(+)Foxp3+] or uninfected [Inf(-)Foxp3+] mice into the tail veins of C57BL/6 mice before the induction of inflammation or during inflammation. Inflammation was induced by ovalbumin (OVA)-alum sensitization and OVA challenge. The concentrations of the Th2-related cytokines IL-4, IL-5, and IL-13 in the bronchial alveolar lavage fluid and the levels of OVA-specific IgE and IgG1 in the serum were lower in mice that received intravenous application of Inf(+)Foxp3+ cells [IV(inf):+(+) group] than in control mice. Some features of allergic airway inflammation were ameliorated by the intravenous application of Inf(-)Foxp3+ cells [IV(inf):+(-) group], but the effects were less distinct than those observed in the IV(inf):+(+) group. We found that Inf(+)Foxp3+ cells migrated to inflammation sites in the lung and expressed higher levels of Treg-cell homing receptors (CCR5 and CCR9) and activation markers (Klrg1, Capg, GARP, Gzmb, OX40) than did Inf(-)Foxp3+ cells. Conclusion/Significance T. spiralis infection promotes the proliferation and functional activation of Treg cells. Parasite-induced Treg cells migrate to the inflammation site and suppress immune responses more effectively than non-parasite-induced Treg cells. The adoptive transfer of Inf(+)Foxp3+ cells is an effective method for the treatment and prevention of allergic airway diseases in mice and is a promising therapeutic approach for the treatment

  7. Lipid droplets fusion in adipocyte differentiated 3T3-L1 cells: A Monte Carlo simulation

    SciTech Connect

    Boschi, Federico; Rizzatti, Vanni; Zamboni, Mauro; Sbarbati, Andrea

    2014-02-15

    Several human worldwide diseases like obesity, type 2 diabetes, hepatic steatosis, atherosclerosis and other metabolic pathologies are related to the excessive accumulation of lipids in cells. Lipids accumulate in spherical cellular inclusions called lipid droplets (LDs) whose sizes range from fraction to one hundred of micrometers in adipocytes. It has been suggested that LDs can grow in size due to a fusion process by which a larger LD is obtained with spherical shape and volume equal to the sum of the progenitors’ ones. In this study, the size distribution of two populations of LDs was analyzed in immature and mature (5-days differentiated) 3T3-L1 adipocytes (first and second populations, respectively) after Oil Red O staining. A Monte Carlo simulation of interaction between LDs has been developed in order to quantify the size distribution and the number of fusion events needed to obtain the distribution of the second population size starting from the first one. Four models are presented here based on different kinds of interaction: a surface weighted interaction (R2 Model), a volume weighted interaction (R3 Model), a random interaction (Random model) and an interaction related to the place where the LDs are born (Nearest Model). The last two models mimic quite well the behavior found in the experimental data. This work represents a first step in developing numerical simulations of the LDs growth process. Due to the complex phenomena involving LDs (absorption, growth through additional neutral lipid deposition in existing droplets, de novo formation and catabolism) the study focuses on the fusion process. The results suggest that, to obtain the observed size distribution, a number of fusion events comparable with the number of LDs themselves is needed. Moreover the MC approach results a powerful tool for investigating the LDs growth process. Highlights: • We evaluated the role of the fusion process in the synthesis of the lipid droplets. • We compared the

  8. Alpha B-crystallin expression in mouse NIH 3T3 fibroblasts: glucocorticoid responsiveness and involvement in thermal protection.

    PubMed Central

    Aoyama, A; Fröhli, E; Schäfer, R; Klemenz, R

    1993-01-01

    alpha B-crystallin, a major soluble protein of vertebrate eye lenses, is a small heat shock protein which transiently accumulates in response to heat shock and other kinds of stress in mouse NIH 3T3 fibroblasts. Ectopic expression of an alpha B-crystallin cDNA clone renders NIH 3T3 cells thermoresistant. alpha B-crystallin accumulates in response to the synthetic glucocorticoid hormone dexamethasone. Dexamethasone-treated NIH 3T3 cells become thermoresistant to the same extent as they accumulate alpha B-crystallin. A cell clone in which alpha B-crystallin is superinduced upon heat shock acquires augmented thermotolerance. Expression of the ras oncogene causes a rapid but transient accumulation of alpha B-crystallin within 1 day. Later, sustained ras oncogene expression suppresses the dexamethasone-mediated alpha B-crystallin accumulation. Thus, oncogenic transformation triggered by the ras oncogene interferes with hormone-mediated accumulation of alpha B-crystallin and concomitant acquisition of thermoresistance. Other known heat shock proteins do not accumulate in response to ectopic alpha B-crystallin expression or to dexamethasone treatment. These results indicate that alpha B-crystallin can protect NIH 3T3 fibroblasts from thermal shock. Images PMID:8441415

  9. 26 CFR 1.67-3T - Allocation of expenses by real estate mortgage investment conduits (temporary).

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 26 Internal Revenue 2 2011-04-01 2011-04-01 false Allocation of expenses by real estate mortgage... Income, Adjusted Gross Income, and Taxable Income § 1.67-3T Allocation of expenses by real estate... real estate mortgage investment conduit or REMIC (as defined in section 860D) shall allocate to each...

  10. 26 CFR 1.67-3T - Allocation of expenses by real estate mortgage investment conduits (temporary).

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 26 Internal Revenue 2 2014-04-01 2014-04-01 false Allocation of expenses by real estate mortgage... Income, Adjusted Gross Income, and Taxable Income § 1.67-3T Allocation of expenses by real estate... real estate mortgage investment conduit or REMIC (as defined in section 860D) shall allocate to each...

  11. Molecular mechanism of 9-cis-retinoic acid inhibition of adipogenesis in 3T3-L1 cells

    SciTech Connect

    Sagara, Chiaki; Takahashi, Katsuhiko; Kagechika, Hiroyuki; Takahashi, Noriko

    2013-03-29

    Highlights: ► We examined the effects of 9-cis-RA on adipogenesis in mouse preadipocyte 3T3-L1. ► 9-cis-RA inhibited lipid accumulation in adipogenetically-induced 3T3-L1 cells. ► A RXR pan-antagonist suppressed the inhibitory effects of 9-cis-RA on adipogenesis. ► This antagonist had no effects on RXRα and PPARγ levels in 9-cis-RA-treated cells. ► 9-cis-RA-induced decrease in both RXRα and PPARγ was independent of RXR activation. -- Abstract: Retinoic acid (RA) signaling is mediated by specific nuclear hormone receptors. Here we examined the effects of 9-cis-RA on adipogenesis in mouse preadipocyte 3T3-L1 cells. 9-cis-RA inhibits the lipid accumulation of adipogenetically induced 3T3-L1 cells. The complex of retinoid X receptor α (RXRα) with peroxisome proliferator-activated receptor γ (PPARγ) is a major transcription factor in the process of adipogenesis, and the levels of these molecules were decreased by 9-cis-RA treatment. A RXR pan-antagonist suppressed 9-cis-RA’s inhibitory effects on adipogenesis, but not on the intracellular levels of both RXRα and PPARγ. These results suggest that 9-cis-RA could inhibit adipogenesis by activating RXR, and decrease both RXR and PPARγs levels in a RXR activation-independent manner.

  12. 26 CFR 1.67-3T - Allocation of expenses by real estate mortgage investment conduits (temporary).

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 26 Internal Revenue 2 2010-04-01 2010-04-01 false Allocation of expenses by real estate mortgage... Income, Adjusted Gross Income, and Taxable Income § 1.67-3T Allocation of expenses by real estate... real estate mortgage investment conduit or REMIC (as defined in section 860D) shall allocate to each...

  13. Technical challenges in 3 T magnetic resonance spectroscopic imaging of the prostate—A single-institution experience

    PubMed Central

    Underwood, Michelle; Boonsirikamchai, Piyaporn; Matin, Surena; Troncoso, Patricia; Ma, Jingfei

    2014-01-01

    The magnetic resonance spectroscopic imaging (MRSI) is the only technique that is currently available in the clinical practice to provide the metabolic status of prostate tissue at the cellular level with a great potential to improve the clinical patient care. Increasing the field strength from 1.5 to 3 T can theoretically provide proportionately higher signal-to-noise ratio (SNR) and improve spectral separation between prostatic metabolite peaks. The technique, however, has been limited to a few academic institutions that are equipped with a team of experts primarily due to due to serious technical challenges in optimizing the spectral quality. High quality shimming is key to the successful MRSI acquisition. Without optimization of the increased field inhomogeneity and radiofrequency (RF) dielectric effect at 3 T, the spectral peak broadening and residual signal from the periprostatic fat tissue may render the overall spectra non-diagnostic. The purpose of this technical note is to present the practical steps of successful acquisition of 3 T MRSI and to address several important technical challenges in minimizing the effect of the increased magnetic field and RF field inhomogeneity in order to obtain highest possible spectral quality based on our initial experience in using 3 T MRSI prototype software. PMID:25202660

  14. Technical challenges in 3 T magnetic resonance spectroscopic imaging of the prostate-A single-institution experience.

    PubMed

    Choi, Haesun; Underwood, Michelle; Boonsirikamchai, Piyaporn; Matin, Surena; Troncoso, Patricia; Ma, Jingfei

    2014-08-01

    The magnetic resonance spectroscopic imaging (MRSI) is the only technique that is currently available in the clinical practice to provide the metabolic status of prostate tissue at the cellular level with a great potential to improve the clinical patient care. Increasing the field strength from 1.5 to 3 T can theoretically provide proportionately higher signal-to-noise ratio (SNR) and improve spectral separation between prostatic metabolite peaks. The technique, however, has been limited to a few academic institutions that are equipped with a team of experts primarily due to due to serious technical challenges in optimizing the spectral quality. High quality shimming is key to the successful MRSI acquisition. Without optimization of the increased field inhomogeneity and radiofrequency (RF) dielectric effect at 3 T, the spectral peak broadening and residual signal from the periprostatic fat tissue may render the overall spectra non-diagnostic. The purpose of this technical note is to present the practical steps of successful acquisition of 3 T MRSI and to address several important technical challenges in minimizing the effect of the increased magnetic field and RF field inhomogeneity in order to obtain highest possible spectral quality based on our initial experience in using 3 T MRSI prototype software. PMID:25202660

  15. 26 CFR 1.367(b)-3T - Repatriation of foreign corporate assets in certain nonrecognition transactions (temporary).

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 26 Internal Revenue 4 2013-04-01 2013-04-01 false Repatriation of foreign corporate assets in...) Effects on Corporation § 1.367(b)-3T Repatriation of foreign corporate assets in certain nonrecognition... (and its direct or indirect owners that would be affected by the election, in the case of an...

  16. 26 CFR 1.367(b)-3T - Repatriation of foreign corporate assets in certain nonrecognition transactions (temporary).

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 26 Internal Revenue 4 2010-04-01 2010-04-01 false Repatriation of foreign corporate assets in... Corporation § 1.367(b)-3T Repatriation of foreign corporate assets in certain nonrecognition transactions... election, the following requirements must be satisfied— (1) The exchanging shareholder (and its direct...

  17. 26 CFR 1.367(b)-3T - Repatriation of foreign corporate assets in certain nonrecognition transactions (temporary).

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 26 Internal Revenue 4 2012-04-01 2012-04-01 false Repatriation of foreign corporate assets in...) Effects on Corporation § 1.367(b)-3T Repatriation of foreign corporate assets in certain nonrecognition... (and its direct or indirect owners that would be affected by the election, in the case of an...

  18. 26 CFR 1.956-3T - Certain trade or service receivables acquired from United States persons (temporary).

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 26 Internal Revenue 10 2010-04-01 2010-04-01 false Certain trade or service receivables acquired... Corporations § 1.956-3T Certain trade or service receivables acquired from United States persons (temporary... includes any trade or service receivable if the trade or service receivable is acquired (directly...

  19. Influence of fat on liver T 1 measurements using modified Look–Locker inversion recovery (MOLLI) methods at 3T

    PubMed Central

    Tunnicliffe, Elizabeth M.; Pavlides, Michael; Robson, Matthew D.

    2016-01-01

    Purpose To characterize the effect of fat on modified Look–Locker inversion recovery (MOLLI) T 1 maps of the liver. The balanced steady‐state free precession (bSSFP) sequence causes water and fat signals to have opposite phase when repetition time (TR) = 2.3 msec at 3T. In voxels that contain both fat and water, the MOLLI T 1 measurement is influenced by the choice of TR. Materials and Methods MOLLI T 1 measurements of the liver were simulated using the Bloch equations while varying the hepatic lipid content (HLC). Phantom scans were performed on margarine phantoms, using both MOLLI and spin echo inversion recovery sequences. MOLLI T 1 at 3T and HLC were determined in patients (n = 8) before and after bariatric surgery. Results At 3T, with HLC in the 0–35% range, higher fat fraction values lead to longer MOLLI T 1 values when TR = 2.3 msec. Patients were found to have higher MOLLI T 1 at elevated HLC (T 1 = 929 ± 97 msec) than at low HLC (T 1 = 870 ± 44 msec). Conclusion At 3T, MOLLI T 1 values are affected by HLC, substantially changing MOLLI T 1 in a clinically relevant range of fat content. J. Magn. Reson. Imaging 2016;44:105–111. PMID:26762615

  20. Ghrelin inhibits the apoptosis of MC3T3-E1 cells through ERK and AKT signaling pathway

    SciTech Connect

    Liang, Qiu-Hua; Liu, Yuan; Wu, Shan-Shan; Cui, Rong-Rong; Yuan, Ling-Qing Liao, Er-Yuan

    2013-11-01

    Ghrelin is a 28-amino-acid peptide that acts as a natural endogenous ligand of the growth hormone secretagogue receptor (GHSR) and strongly stimulates the release of growth hormone from the hypothalamus–pituitary axis. Previous studies have identified the important physiological effects of ghrelin on bone metabolism, such as regulating proliferation and differentiation of osteoblasts, independent of GH/IGF-1 axis. However, research on effects and mechanisms of ghrelin on osteoblast apoptosis is still rare. In this study, we identified expression of GHSR in MC3T3-E1 cells and determined the effects of ghrelin on the apoptosis of osteoblastic MC3T3-E1 cells and the mechanism involved. Our data demonstrated that ghrelin inhibited the apoptosis of osteoblastic MC3T3-E1 cells induced by serum deprivation, as determined by terminal deoxynucleotidyl transferase-mediated deoxyribonucleotide triphosphate nick end-labeling (TUNEL) and ELISA assays. Moreover, ghrelin upregulated Bcl-2 expression and downregulated Bax expression in a dose-dependent manner. Our study also showed decreased activated caspase-3 activity under the treatment of ghrelin. Further study suggested that ghrelin stimulated the phosphorylation of ERK and AKT. Pretreatment of cells with the ERK inhibitor PD98059, PI3K inhibitor LY294002, and GHSR-siRNA blocked the ghrelin-induced activation of ERK and AKT, respectively; however, ghrelin did not stimulate the phosphorylation of p38 or JNK. PD90859, LY294002 and GHSR-siRNA attenuated the anti-apoptosis effect of ghrelin in MC3T3-E1 cells. In conclusion, ghrelin inhibits the apoptosis of osteoblastic MC3T3-E1 cells induced by serum deprivation, which may be mediated by activating the GHSR/ERK and GHSR/PI3K/AKT signaling pathways. - Highlights: • We explored the effects of ghrelin on serum deprivation-induced MC3T3-E1 cells apoptosis. • Both ELISA and TUNEL were used to detect the apoptosis. • The receptor of ghrelin, GHSR, was expressed in MC3T3-E1

  1. Hematopoietic progenitor cells grow on 3T3 fibroblast monolayers that overexpress growth arrest-specific gene-6 (GAS6).

    PubMed

    Dormady, S P; Zhang, X M; Basch, R S

    2000-10-24

    Pluripotential hematopoietic stem cells grow in close association with bone marrow stromal cells, which play a critical role in sustaining hematopoiesis in long-term bone marrow cultures. The mechanisms through which stromal cells act to support pluripotential hematopoietic stem cells are largely unknown. This study demonstrates that growth arrest-specific gene-6 (GAS6) plays an important role in this process. GAS6 is a ligand for the Axl (Ufo/Ark), Sky (Dtk/Tyro3/Rse/Brt/Tif), and Mer (Eyk) family of tyrosine kinase receptors and binds to these receptors via tandem G domains at its C terminus. After translation, GAS6 moves to the lumen of the endoplasmic reticulum, where it is extensively gamma-carboxylated. The carboxylation process is vitamin K dependent, and current evidence suggests that GAS6 must be gamma-carboxylated to bind and activate any of the cognate tyrosine kinase receptors. Here, we show that expression of GAS6 is highly correlated with the capacity of bone marrow stromal cells to support hematopoiesis in culture. Nonsupportive stromal cell lines express little to no GAS6, whereas supportive cell lines express high levels of GAS6. Transfection of the cDNA encoding GAS6 into 3T3 fibroblasts is sufficient to render this previously nonsupportive cell line capable of supporting long-term hematopoietic cultures. 3T3 cells, genetically engineered to stably express GAS6 (GAS6-3T3), produce a stromal layer that supports the generation of colony-forming units in culture (CFU-c) for up to 6 wk. Hematopoietic support by genetically engineered 3T3 is not vitamin K dependent, and soluble recombinant GAS6 does not substitute for coculturing the hematopoietic progenitors with genetically modified 3T3 cells. PMID:11050245

  2. Inhibition of inflammatory signaling pathways in 3T3-L1 adipocytes by apolipoprotein A-I.

    PubMed

    Sultana, Afroza; Cochran, Blake J; Tabet, Fatiha; Patel, Mili; Torres, Luisa Cuesta; Barter, Philip J; Rye, Kerry-Anne

    2016-06-01

    Activation of inflammatory signaling pathways links obesity with metabolic disorders. TLR4-mediated activation of MAPKs and NF-κB are 2 such pathways implicated in obesity-induced inflammation. Apolipoprotein A-I (apoA-I) exerts anti-inflammatory effects on adipocytes by effluxing cholesterol from the cells via the ATP binding cassette transporter A1 (ABCA1). It is not known if these effects involve inhibition of inflammatory signaling pathways by apoA-I. This study asks if apoA-I inhibits activation of MAPKs and NF-κB in mouse 3T3-L1 adipocytes and whether this inhibition is ABCA1 dependent. Incubation of differentiated 3T3-L1 adipocytes with apoA-I decreased cell surface expression of TLR4 by 16 ± 2% and synthesis of the TLR4 adaptor protein, myeloid differentiation primary response 88, by 24 ± 4% in an ABCA1-dependent manner. ApoA-I also inhibited downstream activation of MAPKs, such as ERK, p38MAPK, and JNK, as well as expression of proinflammatory adipokines in bacterial LPS-stimulated 3T3-L1 adipocytes in an ABCA1-dependent manner. ApoA-I, by contrast, suppressed nuclear localization of the p65 subunit of NF-κB by 30 ± 3% in LPS-stimulated 3T3-L1 adipocytes in an ABCA1-independent manner. In conclusion, apoA-I inhibits TLR4-mediated inflammatory signaling pathways in adipocytes by preventing MAPK and NF-κB activation.-Sultana, A., Cochran, B. J., Tabet, F., Patel, M., Cuesta Torres, L., Barter, P. J., Rye, K.-A. Inhibition of inflammatory signaling pathways in 3T3-L1 adipocytes by apolipoprotein A-I. PMID:26965683

  3. Bovine Collagen Peptides Compounds Promote the Proliferation and Differentiation of MC3T3-E1 Pre-Osteoblasts

    PubMed Central

    Liu, JunLi; Zhang, Bing; Song, ShuJun; Ma, Ming; Si, ShaoYan; Wang, YiHu; Xu, BingXin; Feng, Kai; Wu, JiGong; Guo, YanChuan

    2014-01-01

    Objective Collagen peptides (CP) compounds, as bone health supplements, are known to play a role in the treatment of osteoporosis. However, the molecular mechanisms of this process remain unclear. This study aimed to investigate the effects of bovine CP compounds on the proliferation and differentiation of MC3T3-E1 cells. Methods Mouse pre-osteoblast cell line MC3T3-E1 subclone 4 cells were treated with bovine CP compounds. Cell proliferation was analyzed by MTT assays and the cell cycle was evaluated by flow cytometry scanning. Furthermore, MC3T3-E1 cell differentiation was analyzed at the RNA level by real-time PCR and at the protein level by western blot analysis for runt-related transcription factor 2 (Runx2), a colorimetric p-nitrophenyl phosphate assay for alkaline phosphatase (ALP), and ELISA for osteocalcin (OC). Finally, alizarin red staining for mineralization was measured using Image Software Pro Plus 6.0. Results Cell proliferation was very efficient after treatment with different concentrations of bovine CP compounds, and the best concentration was 3 mg/mL. Bovine CP compounds significantly increased the percentage of MC3T3-E1 cells in G2/S phase. Runx2 expression, ALP activity, and OC production were significantly increased after treatment with bovine CP compounds for 7 or 14 days. Quantitative analyses with alizarin red staining showed significantly increased mineralization of MC3T3-E1 cells after treatment with bovine CP compounds for 14 or 21 days. Conclusions Bovine CP compounds increased osteoblast proliferation, and played positive roles in osteoblast differentiation and mineralized bone matrix formation. Taking all the experiments together, our study indicates a molecular mechanism for the potential treatment of osteoarthritis and osteoporosis. PMID:24926875

  4. 6-gingerol prevents adipogenesis and the accumulation of cytoplasmic lipid droplets in 3T3-L1 cells.

    PubMed

    Tzeng, Thing-Fong; Liu, I-Min

    2013-04-15

    6-Gingerol ((S)-5-hydroxy-1-(4-hydroxy-3-methoxyphenyl)-3-decanone) is one of the pungent constituents of Zingiber zerumbet (L) Smith (Zingiberaceae family). In this study, we investigated the effects of 6-gingerol on the inhibition of adipogenesis in 3T3-L1 cells. After treatment with 6-gingerol in differentiation medium for 4 or 8 days, the 3T3-L1 cells were lysed for experimental analysis. Cells were stained with Oil-Red-O to detect oil droplets in adipocytes. The 3T3-L1 cells were lysed and measured for triglyceride contents. The protein expression of adipogenesis-related transcription factor was evaluated by Western blot analysis. 6-Gingerol suppressed oil droplet accumulation and reduced the droplet size in a concentration (5-15 μg/ml)- and time-dependent manner. Treatment of 3T3-L1 cells with 6-gingerol reduced the protein levels of peroxisome proliferator-activated receptor (PPAR)γ and CCAAT/enhancer-binding protein (C/EBP)α. Additionally, the protein levels of fatty acid synthase (FAS) and adipocyte-specific fatty acid binding protein (aP2) decreased upon treatment with 6-gingerol. Meanwhile, 6-gingerol diminished the insulin-stimulated serine phosphorylation of Akt (Ser473) and GSK3β (Ser9). These results suggest that 6-gingerol effectively suppresses adipogenesis and that it exerts its role mainly through the significant down-regulation of PPARγ and C/EBPα and subsequently inhibits FAS and aP2 expression. 6-Gingerol also inhibited differentiation in 3T3-L1 cells by attenuating the Akt/GSK3β pathway. Our findings provide important insights into the mechanisms underlying the anti-adipogenic activity of 6-gingerol. PMID:23369342

  5. Image quality and signal distribution in 1.5-T and 3-T MRI in mild traumatic brain injury patients

    NASA Astrophysics Data System (ADS)

    Rossi, Maija E.; Dastidar, Prasun; Ryymin, Pertti; Ylinen, Aarne; Öhman, Juha; Soimakallio, Seppo; Eskola, Hannu

    2009-02-01

    Clear standards are lacking in the imaging modalities of the deficit in mild traumatic brain injury (MTBI) patients. The purpose of this study is to compare the image quality by signal distribution between 1.5 Tesla and 3 Tesla MRI in turbo spin echo (TSE) and gradient echo (GRE) images in normal hospital settings and to find preferences for which field to use in MTBI patients. We studied 40 MTBI patients with TSE and GRE; 20 patients were imaged at 1.5 T and 20 at 3 T. The imaging parameters were optimized separately for the two scanners. Histograms of the signal distribution in 22 ROIs were fitted to a 1-peak Gaussian model and the resulting peak positions were scaled in respect to the peak positions of genu of the corpus callosum and the caudate nuclei. Correlation of the contrast of the ROIs in reference to genu of the corpus callosum between both the two scanners and the two imaging sequences was good. Image contrast was similar at both in the TSE images; in the GRE images contrast improved from 1.5 T to 3 T. However, based on peak positions and widths, a slight drawback in the separability between the ROIs was observed when 1.5 T MRI was replaced by 3 T. No clear improvement in tissue contrast or separability of 3 T was found compared to 1.5 T. Imaging of MTBI with 3 T should therefore be based on other advantages of high-field imaging, such as improved SNR and spatial resolution.

  6. Bipolar I Disorder and Schizophrenia: A 440–Single-Nucleotide Polymorphism Screen of 64 Candidate Genes among Ashkenazi Jewish Case-Parent Trios

    PubMed Central

    Fallin, M. Daniele ; Lasseter, Virginia K. ; Avramopoulos, Dimitrios ; Nicodemus, Kristin K. ; Wolyniec, Paula S. ; McGrath, John A. ; Steel, Gary ; Nestadt, Gerald ; Liang, Kung-Yee ; Huganir, Richard L. ; Valle, David ; Pulver, Ann E. 

    2005-01-01

    Bipolar, schizophrenia, and schizoaffective disorders are common, highly heritable psychiatric disorders, for which familial coaggregation, as well as epidemiological and genetic evidence, suggests overlapping etiologies. No definitive susceptibility genes have yet been identified for any of these disorders. Genetic heterogeneity, combined with phenotypic imprecision and poor marker coverage, has contributed to the difficulty in defining risk variants. We focused on families of Ashkenazi Jewish descent, to reduce genetic heterogeneity, and, as a precursor to genomewide association studies, we undertook a single-nucleotide polymorphism (SNP) genotyping screen of 64 candidate genes (440 SNPs) chosen on the basis of previous linkage or of association and/or biological relevance. We genotyped an average of 6.9 SNPs per gene, with an average density of 1 SNP per 11.9 kb in 323 bipolar I disorder and 274 schizophrenia or schizoaffective Ashkenazi case-parent trios. Using single-SNP and haplotype-based transmission/disequilibrium tests, we ranked genes on the basis of strength of association (P<.01). Six genes (DAO, GRM3, GRM4, GRIN2B, IL2RB, and TUBA8) met this criterion for bipolar I disorder; only DAO has been previously associated with bipolar disorder. Six genes (RGS4, SCA1, GRM4, DPYSL2, NOS1, and GRID1) met this criterion for schizophrenia or schizoaffective disorder; five replicate previous associations, and one, GRID1, shows a novel association with schizophrenia. In addition, six genes (DPYSL2, DTNBP1, G30/G72, GRID1, GRM4, and NOS1) showed overlapping suggestive evidence of association in both disorders. These results may help to prioritize candidate genes for future study from among the many suspected/proposed for schizophrenia and bipolar disorders. They provide further support for shared genetic susceptibility between these two disorders that involve glutamate-signaling pathways. PMID:16380905

  7. Bipolar I disorder and schizophrenia: a 440-single-nucleotide polymorphism screen of 64 candidate genes among Ashkenazi Jewish case-parent trios.

    PubMed

    Fallin, M Daniele; Lasseter, Virginia K; Avramopoulos, Dimitrios; Nicodemus, Kristin K; Wolyniec, Paula S; McGrath, John A; Steel, Gary; Nestadt, Gerald; Liang, Kung-Yee; Huganir, Richard L; Valle, David; Pulver, Ann E

    2005-12-01

    Bipolar, schizophrenia, and schizoaffective disorders are common, highly heritable psychiatric disorders, for which familial coaggregation, as well as epidemiological and genetic evidence, suggests overlapping etiologies. No definitive susceptibility genes have yet been identified for any of these disorders. Genetic heterogeneity, combined with phenotypic imprecision and poor marker coverage, has contributed to the difficulty in defining risk variants. We focused on families of Ashkenazi Jewish descent, to reduce genetic heterogeneity, and, as a precursor to genomewide association studies, we undertook a single-nucleotide polymorphism (SNP) genotyping screen of 64 candidate genes (440 SNPs) chosen on the basis of previous linkage or of association and/or biological relevance. We genotyped an average of 6.9 SNPs per gene, with an average density of 1 SNP per 11.9 kb in 323 bipolar I disorder and 274 schizophrenia or schizoaffective Ashkenazi case-parent trios. Using single-SNP and haplotype-based transmission/disequilibrium tests, we ranked genes on the basis of strength of association (P<.01). Six genes (DAO, GRM3, GRM4, GRIN2B, IL2RB, and TUBA8) met this criterion for bipolar I disorder; only DAO has been previously associated with bipolar disorder. Six genes (RGS4, SCA1, GRM4, DPYSL2, NOS1, and GRID1) met this criterion for schizophrenia or schizoaffective disorder; five replicate previous associations, and one, GRID1, shows a novel association with schizophrenia. In addition, six genes (DPYSL2, DTNBP1, G30/G72, GRID1, GRM4, and NOS1) showed overlapping suggestive evidence of association in both disorders. These results may help to prioritize candidate genes for future study from among the many suspected/proposed for schizophrenia and bipolar disorders. They provide further support for shared genetic susceptibility between these two disorders that involve glutamate-signaling pathways. PMID:16380905

  8. Measurement of the modulation transfer function of an X-ray microscope based on multiple Fourier orders analysis of a Siemens star.

    PubMed

    Otón, Joaquín; Sorzano, Carlos Oscar S; Marabini, Roberto; Pereiro, Eva; Carazo, Jose M

    2015-04-20

    Soft X-ray tomography (SXT) is becoming a powerful imaging technique to analyze eukaryotic whole cells close to their native state. Central to the analysis of the quality of SXT 3D reconstruction is the estimation of the spatial resolution and Depth of Field of the X-ray microscope. In turn, the characterization of the Modulation Transfer Function (MTF) of the optical system is key to calculate both parameters. Consequently, in this work we introduce a fully automated technique to accurately estimate the transfer function of such an optical system. Our proposal is based on the preprocessing of the experimental images to obtain an estimate of the input pattern, followed by the analysis in Fourier space of multiple orders of a Siemens Star test sample, extending in this way its measured frequency range. PMID:25968993

  9. Nucleosome formation with the testis-specific histone H3 variant, H3t, by human nucleosome assembly proteins in vitro

    PubMed Central

    Tachiwana, Hiroaki; Osakabe, Akihisa; Kimura, Hiroshi; Kurumizaka, Hitoshi

    2008-01-01

    Five non-allelic histone H3 variants, H3.1, H3.2, H3.3, H3t and CENP-A, have been identified in mammals. H3t is robustly expressed in the testis, and thus was assigned as the testis-specific H3 variant. However, recent proteomics and tissue-specific RT-PCR experiments revealed a small amount of H3t expression in somatic cells. In the present study, we purified human H3t as a recombinant protein, and showed that H3t/H4 forms nucleosomes with H2A/H2B by the salt-dialysis method, like the conventional H3.1/H4. We found that H3t/H4 is not efficiently incorporated into the nucleosome by human Nap1 (hNap1), due to its defective H3t/H4 deposition on DNA. In contrast, human Nap2 (hNap2), a paralog of hNap1, promotes nucleosome assembly with H3t/H4. Mutational analyses revealed that the Ala111 residue, which is conserved among H3.1, H3.2 and H3.3, but not in H3t, is the essential residue for the hNap1-mediated nucleosome assembly. These results suggest that H3t may be incorporated into chromatin by a specific chaperone-mediated pathway. PMID:18281699

  10. Osmotically inducible uptake of betaine via amino acid transport system A in SV-3T3 cells.

    PubMed

    Petronini, P G; De Angelis, E; Borghetti, A F; Wheeler, K P

    1994-05-15

    The osmotically inducible uptake of betaine (NNN-trimethylglycine) by SV-3T3 cells has been studied and compared with the similar process in MDCK cells. Betaine uptake by SV-3T3 cells could be described in terms of a saturable, Na(+)-dependent, component plus a small non-saturable, Na(+)-independent, component. Transport was active, producing considerable accumulation of betaine in the cells. After exposure of the cells to hypertonic conditions for 6 h, there was a marked increase in betaine uptake. Kinetic analysis indicated that this increase resulted from an increase in the Vmax. value of the saturable component, from about 88 to 185 nmol of betaine/5 min per mg of protein, the corresponding Km values of about 15 and 10 mM not being significantly different. This induction of transport activity was detectable only after about 2 h exposure of the cells to hypertonic medium, closely paralleling an induction of influx of N-methylaminoisobutyric acid, and was prevented by the presence of cycloheximide. Betaine influx was markedly inhibited by several neutral amino acids, particularly those transported by system A, such as N-methylaminoisobutyric acid and the imino acid proline. A high concentration (25 mM) of betaine also significantly inhibited the uptake of proline by SV-3T3 cells. Although very similar results were obtained with MDCK cells, prolonged exposure of cells to hypertonic conditions revealed distinct differences. When the hypertonic incubation was extended from 6 h to 24 h, betaine transport in SV-3T3 cells either remained the same or decreased, whereas it showed a further marked increase in MDCK cells, and also became sensitive to inhibition by gamma-aminobutyric acid. mRNA for the betaine transporter BGT-1 [Yamauchi, Uchida, Kwon, Preston, Brooks Robey, Garcia-Perez, Burg and Handler (1992) J. Biol. Chem. 267, 649-652] was detectable in MDCK cells exposed to hypertonic medium for 24 h, but not in SV-3T3 cells under any conditions. It is concluded that

  11. The effects of ascorbic acid and iron co-supplementation on the proliferation of 3T3 fibroblasts.

    PubMed

    Collis, C S; Yang, M; Peach, S J; Diplock, A T; Rice-Evans, C

    1996-07-01

    Exposure of 3T3 fibroblasts to FeII reveals a concentration-dependent inhibition of cell proliferation compared to control cells, the apparent threshold for this iron-mediated effect being 5 microM FeII. The inhibition of cell proliferation was accompanied by an enhancement of total malondialdehyde (MDA) levels (as detected directly by hplc) in the cells at higher iron concentrations. The co-supplementation of FeII with varying concentrations of ascorbic acid over the range 5 microM to 240 microM had no significant effect on the threshold for iron toxicity or lipid peroxidation. These results show that there is neither a significant exacerbation of the pro-oxidant effect of FeII nor any protective effect of ascorbate when cultures of 3T3 mouse fibroblasts are exposed to co-supplementation regimes of iron with ascorbic acid. PMID:8814446

  12. FGF-2 signaling induces downregulation of TAZ protein in osteoblastic MC3T3-E1 cells

    SciTech Connect

    Eda, Homare; Aoki, Katsuhiko; Marumo, Keishi; Fujii, Katsuyuki; Ohkawa, Kiyoshi

    2008-02-08

    Transcriptional coactivator with PDZ-binding motif (TAZ) protein is a coactivator of Runx2 and corepressor of PPAR{gamma}. It also induces differentiation of mesenchymal cells into osteoblasts. In this study, we found that FGF-2, which inhibits bone mineralization and stimulates cell proliferation, reduced the TAZ protein expression level in osteoblast-like cells, MC3T3-E1. This reduction was recovered by removing FGF-2 from the culture medium, which also restored the osteoblastic features of MC3T3-E1 cells. Furthermore, FGF-2-induced reduction of TAZ is blocked by a SAPK/JNK-specific inhibitor. These findings suggest that the expression of TAZ protein is involved in osteoblast proliferation and differentiation. This may help elucidate the discrepancies in the effect of FGF-2 and contribute to the understanding of FGF/FGFR-associated craniosynostosis syndrome etiology and treatment.

  13. Ectopic osteogenic tissue formation by MC3T3-E1 cell-laden chitosan/hydroxyapatite composite scaffold.

    PubMed

    Koç, Aysel; Elçin, Ayşe Eser; Elçin, Yaşar Murat

    2016-09-01

    This study evaluates the suitability of a macroporous three-dimensional chitosan/hydroxyapatite (CS/HA) composite as a bone tissue engineering scaffold using MC3T3-E1 cells. The CS/HA scaffold was produced by freeze-drying, and characterized by means of SEM and FTIR. In vitro findings demonstrated that CS/HA supported attachment and proliferation of cells, and stimulated extracellular matrix (ECM) production. Tissue biocompatibility and osteogenic capacity of the cell-laden constructs were evaluated in an ectopic Wistar rat model. In vivo results showed that the MC3T3-E1 cell-laden CS/HA was essentially histocompatible, promoted neovascularization and calcified matrix formation, and secreted osteoblast-specific protein. We conclude that the composite scaffold evaluated has potential for applications in bone regeneration. PMID:25968048

  14. Deoxyactein Isolated from Cimicifuga racemosa protects osteoblastic MC3T3-E1 cells against antimycin A-induced cytotoxicity.

    PubMed

    Choi, Eun Mi

    2013-06-01

    Deoxyactein is one of the major constituents isolated from Cimicifuga racemosa. In the present study, we investigated the protective effects of deoxyactein on antimycin A (mitochondrial electron transport inhibitor)-induced toxicity in osteoblastic MC3T3-E1 cells. Exposure of MC3T3-E1 cells to antimycin A caused significant cell viability loss, as well as mitochondrial membrane potential dissipation, complex IV inactivation, ATP loss, intracellular calcium ([Ca(2+) ]i ) elevation and oxidative stress. Pretreatment with deoxyactein prior to antimycin A exposure significantly reduced antimycin A-induced cell damage by preventing mitochondrial membrane potential dissipation, complex IV inactivation, ATP loss, [Ca(2+) ]i elevation and oxidative stress. Moreover, deoxyactein increased the activation of PI3K (phosphoinositide 3-kinase), Akt (protein kinase B) and CREB (cAMP-response element-binding protein) inhibited by antimycin A. All these data indicate that deoxyactein may reduce or prevent osteoblasts degeneration in osteoporosis or other degenerative disorders. PMID:22180388

  15. Long-term exposure of 3T3 fibroblast cells to endocrine disruptors alters sensitivity to oxidative injury.

    PubMed

    Nishimura, Yuka; Nakai, Yasuyoshi; Tanaka, Aiko; Nagao, Tetsuji; Fukushima, Nobuyuki

    2014-07-01

    When Swiss 3T3 fibroblasts were exposed to bisphenol A (BPA) or nonylphenol (NP) within a range of 0.1-100 nM for 30-45 days, increased resistance to oxidative injury was found. Western blot analysis indicated concomitant increased expression of bcl-2 protein and reduced histone methylation levels in cells after BPA or NP exposure. Using a heterologous expression system, both chemicals could stimulate G protein-coupled receptor 30 (GPR30), a transmembrane estrogen receptor predominantly expressed in 3T3 cells, at lower concentrations, which gave increased survival. Taken together, these results suggest that BPA or NP exposure might cause alterations in cellular activity against oxidative stress, possibly through GPR30. PMID:24604882

  16. Bombesin stimulation of c-fos and c-myc gene expression in cultured of Swiss 3T3 cells

    SciTech Connect

    Palumbo, A.P.; Rossino, P.; Comoglio, P.M.

    1986-11-01

    Bombesin has been show to be a potent mitogen for Swiss 3T3 cells. At nanomolar concentrations it stimulates DNA synthesis in quiescent cultures of 3T3 cells and also induces the expression of c-fos and c-myc mRNA. c-fos mRNA transcripts dramatically increase 15 min after the addition of bombesin, are still abundant after 30-60 min and then decrease. c-myc mRNA induction is detectable later, 1 h after bombesin treatment. Conversely, no changes in c-Ki-ras expression are observed after stimulation with bombesin. These results demonstrate that the increased expression of c-fos and c-myc mRNAs appears to be a common response to diverse agents that induce DNA synthesis and cell proliferation.

  17. Global gene expression profiling of JMJD6- and JMJD4-depleted mouse NIH3T3 fibroblasts

    PubMed Central

    Hu, Yu-Jie; Imbalzano, Anthony N.

    2016-01-01

    Emerging evidence suggests Jumonji domain-containing proteins are epigenetic regulators in diverse biological processes including cellular differentiation and proliferation. RNA interference-based analyses combined with gene expression profiling can effectively characterize the cellular functions of these enzymes. We found that the depletion of Jumonji domain-containing protein 6 (JMJD6) and its paralog protein Jumonji domain-containing protein 4 (JMJD4) individually by small hairpin RNAs (shRNAs) slowed cell proliferation of mouse NIH3T3 fibroblasts. We subsequently performed gene expression profiling on both JMJD6- and JMJD4-depleted mouse NIH3T3 fibroblasts using the Affymetrix GeneChip Mouse Exon 1.0 ST Array. Here we report the gene profiling datasets along with the experimental procedures. The information can be used to further investigate how JMJD6 and JMJD4 affect gene expression and cellular physiology. PMID:27071056

  18. Magnetic Levitation of MC3T3 Osteoblast Cells as a Ground-Based Simulation of Microgravity.

    PubMed

    Hammer, Bruce E; Kidder, Louis S; Williams, Philip C; Xu, Wayne Wenzhong

    2009-11-01

    Diamagnetic samples placed in a strong magnetic field and a magnetic field gradient experience a magnetic force. Stable magnetic levitation occurs when the magnetic force exactly counter balances the gravitational force. Under this condition, a diamagnetic sample is in a simulated microgravity environment. The purpose of this study is to explore if MC3T3-E1 osteoblastic cells can be grown in magnetically simulated hypo-g and hyper-g environments and determine if gene expression is differentially expressed under these conditions. The murine calvarial osteoblastic cell line, MC3T3-E1, grown on Cytodex-3 beads, were subjected to a net gravitational force of 0, 1 and 2 g in a 17 T superconducting magnet for 2 days. Microarray analysis of these cells indicated that gravitational stress leads to up and down regulation of hundreds of genes. The methodology of sustaining long-term magnetic levitation of biological systems are discussed. PMID:20052306

  19. Tumor slices as a model to evaluate doxorubicin in vitro treatment and expression of trios of genes PRSS11, MTSS1, CLPTM1 and PRSS11, MTSS1, SMYD2 in canine mammary gland cancer

    PubMed Central

    Sobral, Renata A; Honda, Suzana T; Katayama, Maria Lucia H; Brentani, Helena; Brentani, M Mitzi; Patrão, Diogo FC; Folgueira, Maria Aparecida AK

    2008-01-01

    Background In women with breast cancer submitted to neoadjuvant chemotherapy based in doxorubicin, tumor expression of groups of three genes (PRSS11, MTSS1, CLPTM1 and PRSS11, MTSS1, SMYD2) have classified them as responsive or resistant. We have investigated whether expression of these trios of genes could predict mammary carcinoma response in dogs and whether tumor slices, which maintain epithelial-mesenchymal interactions, could be used to evaluate drug response in vitro. Methods Tumors from 38 dogs were sliced and cultured with or without doxorubicin 1 μM for 24 h. Tumor cells were counted by two observers to establish a percentage variation in cell number, between slices. Based on these results, a reduction in cell number between treated and control samples ≥ 21.7%, arbitrarily classified samples, as drug responsive. Tumor expression of PRSS11, MTSS1, CLPTM1 and SMYD2, was evaluated by real time PCR. Relative expression results were then transformed to their natural logarithm values, which were spatially disposed according to the expression of trios of genes, comprising PRSS11, MTSS1, CLPTM1 and PRSS11, MTSS1, SMYD2. Fisher linear discrimination test was used to generate a separation plane between responsive and non-responsive tumors. Results Culture of tumor slices for 24 h was feasible. Nine samples were considered responsive and 29 non-responsive to doxorubicin, considering the pre-established cut-off value of cell number reduction ≥ 21.7%, between doxorubicin treated and control samples. Relative gene expression was evaluated and tumor samples were then spatially distributed according to the expression of the trios of genes: PRSS11, MTSS1, CLPTM1 and PRSS11, MTSS1, SMYD2. A separation plane was generated. However, no clear separation between responsive and non-responsive samples could be observed. Conclusion Three-dimensional distribution of samples according to the expression of the trios of genes PRSS11, MTSS1, CLPTM1 and PRSS11, MTSS1, SMYD2 could

  20. Hydroxytyrosol Inhibits Cannabinoid CB1 Receptor Gene Expression in 3T3-L1 Preadipocyte Cell Line.

    PubMed

    Tutino, Valeria; Orlando, Antonella; Russo, Francesco; Notarnicola, Maria

    2016-02-01

    The 3T3-L1 preadipocyte cell line is a well characterized cell model for studying the adipocyte status and the molecular mechanisms involved in differentiation of these cells. 3T3-L1 preadipocytes have the ability to synthesize and degrade endocannabinoid anandamide (AEA) and their differentiation into adipocytes increases the expression of cannabinoid (CB1) and PPAR-γ receptors. Clinically, the blocking stimulation of the endocannabinoid pathway has been one of the first approaches proposed to counteract the obesity and obesity-associated diseases (such as diabetes, metabolic syndrome and cancer). In this connection, here we studied in cultured 3T3-L1 pre-adipocytes the effects of n-3-PUFA, α-Linolenic acid (OM-3), n-6-PUFA, Linoleic acid (OM-6), and hydroxytyrosol (HT) on the expression of CB1 receptor gene and the adipogenesis-related genes PPAR-γ, Fatty Acid Synthase (FAS) and Lipoprotein Lipase (LPL). HT was able to inhibit 3T3-L1 cell differentiation by down-regulating cell proliferation and CB1 receptor gene expression. HT exhibited anti-adipogenic effects, whereas OM-3 and OM-6 exerted an inhibitory action on cell proliferation associated with an induction of the preadipocytes differentiation and CB1 receptor gene expression. Moreover, the expression of FAS and LPL genes resulted increased after treatment with both HT and OM-3 and OM-6. The present study points out that the intake of molecules such as HT, contained in extra virgin olive oil, may be considered also in view of antiobesity and antineoplastic properties by acting directly on the adipose tissue and modulating CB1 receptor gene transcription. PMID:26189725

  1. Inhibition of adipogenesis and leptin production in 3T3-L1 adipocytes by a derivative of meridianin C

    SciTech Connect

    Park, Yu-Kyoung; Lee, Tae-Yoon; Choi, Jong-Soon; Hong, Victor Sukbong; Lee, Jinho; Park, Jong-Wook; Jang, Byeong-Churl

    2014-10-03

    Highlights: • Compound 7b, a meridianin C derivative, inhibits adipogenesis. • Compound 7b inhibits C/EBP-α, PPAR-γ, FAS, STAT-3, and STAT-5 in 3T3-L1 adipocytes. • Compound 7b inhibits leptin, but not adiponectin, expression in 3T3-L1 adipocytes. • Compound 7b thus may have therapeutic potential against obesity. - Abstract: Meridianin C, a marine alkaloid, is a potent protein kinase inhibitor and has anti-cancer activity. We have recently developed a series of meridianin C derivatives (compound 7a–7j) and reported their proviral integration Moloney Murine Leukemia Virus (pim) kinases’ inhibitory and anti-proliferative effects on human leukemia cells. Here we investigated the effect of these meridianin C derivatives on adipogenesis. Strikingly, among the derivatives tested, compound 7b most strongly inhibited lipid accumulation during the differentiation of 3T3-L1 preadipocytes into adipocytes. However, meridianin C treatment was largely cytotoxic to 3T3-L1 adipocytes. On mechanistic levels, compound 7b reduced not only the expressions of CCAAT/enhancer-binding protein-α (C/EBP-α), peroxisome proliferator-activated receptor-γ (PPAR-γ), and fatty acid synthase (FAS) but also the phosphorylation levels of signal transducer and activator of transcription-3 (STAT-3) and STAT-5 during adipocyte differentiation. Moreover, compound 7b repressed leptin, but not adiponectin, expression during adipocyte differentiation. Collectively, these findings demonstrate that a meridianin C derivative inhibits adipogenesis by down-regulating expressions and/or phosphorylations of C/EBP-α, PPAR-γ, FAS, STAT-3 and STAT-5.

  2. Cloning and Expression of CD19, a Human B-Cell Marker in NIH-3T3 Cell Line

    PubMed Central

    Abbasi-Kenarsari, Hajar; Shafaghat, Farzaneh; Baradaran, Behzad; Movassaghpour, Ali Akbar; Shanehbandi, Dariush; Kazemi, Tohid

    2015-01-01

    Background CD19 is a pan B cell marker that is recognized as an attractive target for antibody-based therapy of B-cell disorders including autoimmune disease and hematological malignancies. The object of this study was to stably express the human CD19 antigen in the murine NIH-3T3 cell line aimed to be used as an immunogen in our future study. Methods Total RNA was extracted from Raji cells in which high expression of CD19 was confirmed by flow cytometry. Synthesized cDNA was used for CD19 gene amplification by conventional PCR method using Pfu DNA polymerase. PCR product was ligated to pGEM-T Easy vector and ligation mixture was transformed to DH5α competent bacteria. After blue/white selection, one positive white colony was subjected to plasmid extraction and direct sequencing. Then, CD19 cDNA was sub-cloned into pCMV6-Neo expression vector by double digestion using KpnI and HindIII enzymes. NIH-3T3 mouse fibroblast cell line was subsequently transfected by the construct using Jet-PEI transfection reagent. After 48 hours, surface expression of CD19 was confirmed by flow cytometry and stably transfected cells were selected by G418 antibiotic. Results Amplification of CD19 cDNA gave rise to 1701 bp amplicon confirmed by alignment to reference sequence in NCBI database. Flow cytometric analysis showed successful transient and stable expression of CD19 on NIH-3T3 cells (29 and 93%, respectively). Conclusion Stable cell surface expression of human CD19 antigen in a murine NIH-3T3 cell line may develop a proper immunogene which raises specific anti-CD19 antibody production in the mice immunized sera. PMID:25926951

  3. Effects of C-reactive protein on adipokines genes expression in 3T3-L1 adipocytes

    SciTech Connect

    Yuan, Guoyue; Jia, Jue; Di, Liangliang; Zhou, Libin; Dong, Sijing; Ye, Jingjing; Wang, Dong; Yang, Ling; Wang, Jifang; Li, Lianxi; Yang, Ying; Mao, Chaoming; Chen, Mingdao

    2012-08-03

    Highlights: Black-Right-Pointing-Pointer CRP increases TNF-{alpha} and IL-6 genes expression in matured 3T3-L1 adipocytes. Black-Right-Pointing-Pointer CRP suppresses adiponectin, leptin and PPAR-{gamma} mRNA levels in matured 3T3-L1 cells. Black-Right-Pointing-Pointer Wortmannin reverses effects of CRP on adiponectin, TNF-{alpha} and leptin mRNA levels. Black-Right-Pointing-Pointer CRP may regulate IR, obesity and metabolic syndrome by this mechanism. -- Abstract: Adipose tissue is now recognized to be an important endocrine organ, secreting a variety of adipokines that are involved in the regulation of energy metabolism, insulin resistance and metabolic syndrome. C-reactive protein (CRP) is considered as one of the most sensitive markers of inflammation. A number of studies have shown that elevation of CRP concentrations is an independent predictive parameter of type 2 diabetes mellitus, which is also strongly associated with various components of the metabolic syndrome. The aim of the present study is to investigate the effects of CRP on adipokines genes expression in 3T3-L1 adipocytes. Quantitative real-time PCR analysis revealed that CRP inhibited adiponectin, leptin and peroxisome proliferator-activated receptor-gamma (PPAR-{gamma}) genes expression and raised tumor necrosis factor-{alpha} (TNF-{alpha}) and interleukin-6 (IL-6) mRNA levels in matured 3T3-L1 adipocytes in a dose and time-dependent manner. Pharmacological inhibition of phosphatidylinositol (PI)-3 kinase by wortmannin partially reversed the effects of CRP on adiponectin, TNF-{alpha} and leptin genes expression. These results collectively suggest that CRP regulates adiponectin, TNF-{alpha}, leptin, IL-6 and PPAR-{gamma} genes expression, and that might represent a mechanism by which CRP regulates insulin resistance, obesity and metabolic syndrome.

  4. A comparison of distributional considerations with statistical analysis of resting state fMRI at 3T and 7T

    NASA Astrophysics Data System (ADS)

    Yang, Xue; Holmes, Martha J.; Newton, Allen T.; Morgan, Victoria L.; Landman, Bennett A.

    2012-02-01

    Ultra-high field 7T magnetic resonance imaging (MRI) offers potentially unprecedented spatial resolution of functional activity within the human brain through increased signal and contrast to noise ratios over traditional 1.5T and 3T MRI scanners. However, the effects physiological and imaging artifacts are also greatly increased. Traditional statistical parametric mapping theories based on distributional properties representative of data acquired at lower fields may be inadequate for new 7T data. Herein, we investigate the model fitting residuals based on two 7T and one 3T protocols. We find that model residuals are substantively more non-Gaussian at 7T relative to 3T. Imaging slices that passed through regions with peak inhomogeneity problems (e.g., mid-brain acquisitions for the 7T hippocampus) exhibited visually higher degrees of distortion along with spatially correlated and extreme values of kurtosis (a measure of non- Gaussianity). The impacts of artifacts have been previously addressed for 3T data by estimating the covariance matrix of the regression errors. We further extend the robust estimation approach for autoregressive models and evaluate the qualitative impacts of this technique relative to traditional inference. Clear differences in statistical significance are shown between inferences based on classical versus robust assumptions, which suggest that inferences based on Gaussian assumptions are subject to practical (as well as theoretical) concerns regarding their power and validity. Hence, modern statistical approaches, such as the robust autoregressive model posed herein, are appropriate and suitable for inference with ultra-high field functional magnetic resonance imaging.

  5. Effects of Pueraria lobata Root Ethanol Extract on Adipogenesis and Lipogenesis During 3T3-L1 Differentiation into Adipocytes.

    PubMed

    Lee, Chae Myoung; Yoon, Mi Sook; Kim, Young Chul

    2015-06-01

    We evaluated the inhibitory effect of Pueraria lobata root ethanol extract (PLREE) on lipid accumulation during 3T3-L1 differentiation to adipocytes by measuring the intracellular expression of adipogenic, lipogenic, and lipolytic markers and lipid accumulation. The total polyphenol and flavonoid content of PLREE were 47 and 29 mg/g, respectively. The electron donating capacity of PLREE at 1,000 μg/mL was 48.8%. Treatment of 3T3-L1 preadipocytes with 100, 250, or 500 μg/mL PLREE for 8 days dose-dependently promoted the differentiation of 3T3-L1 cells. In contrast, the lipid content of PLREE-treated cells was significantly reduced by 7.8% (p < 0.05), 35.6% (p < 0.001), and 42.2% (p < 0.001) following treatment with 100, 250, and 500 μg/mL PLREE, respectively, as compared to differentiated control cells. PLREE upregulated peroxisome proliferator-activated receptor γ mRNA and protein, and sterol regulator element-binding protein-1c mRNA levels, but did not affect CCAAT/enhancer binding-protein β and α mRNA levels. PLREE also downregulated acetyl-CoA carboxylase mRNA and protein, fatty acid synthase (FAS) protein, and leptin mRNA levels, but did not affect FAS mRNA expression. PLREE upregulated adipose triglyceride lipase mRNA and protein expression, and hormone-sensitive lipase (HSL) protein expression, but did not affect HSL mRNA expression. In conclusion, we found that PLREE enhanced adipogenesis, but reduced lipogenesis, resulting in decreased lipid accumulation in 3T3-L1 cells. PMID:26191386

  6. Photo catalogue for the classification of foci in the BALB/c 3T3 cell transformation assay.

    PubMed

    Sasaki, Kiyoshi; Bohnenberger, Susanne; Hayashi, Kumiko; Kunkelmann, Thorsten; Muramatsu, Dai; Poth, Albrecht; Sakai, Ayako; Salovaara, Susan; Tanaka, Noriho; Thomas, B Claire; Umeda, Makoto

    2012-04-11

    This catalogue is a display of focus photos representative of the BALB/c 3T3 cell transformation assay (CTA). It is intended as a visual aid for the identification and the scoring of foci in the conduct of the assay. A proper training from experienced personnel together with the protocol reported in this issue and the present photo catalogue will support method transfer and consistency in the assay results. PMID:22331008

  7. Functional expression of 5-HT{sub 2A} receptor in osteoblastic MC3T3-E1 cells

    SciTech Connect

    Hirai, Takao; Kaneshige, Kota; Kurosaki, Teruko; Nishio, Hiroaki

    2010-05-28

    In the previous study, we reported the gene expression for proteins related to the function of 5-hydroxytryptamine (5-HT, serotonin) and elucidated the expression patterns of 5-HT{sub 2} receptor subtypes in mouse osteoblasts. In the present study, we evaluated the possible involvement of 5-HT receptor subtypes and its inactivation system in MC3T3-E1 cells, an osteoblast cell line. DOI, a 5-HT{sub 2A} and 5-HT{sub 2C} receptor selective agonist, as well as 5-HT concentration-dependently increased proliferative activities of MC3T3-E1 cells in their premature period. This effect of 5-HT on cell proliferation were inhibited by ketanserin, a 5-HT{sub 2A} receptor specific antagonist. Moreover, both DOI-induced cell proliferation and phosphorylation of ERK1 and 2 proteins were inhibited by PD98059 and U0126, selective inhibitors of MEK in a concentration-dependent manner. Furthermore, treatment with fluoxetine, a 5-HT specific re-uptake inhibitor which inactivate the function of extracellular 5-HT, significantly increased the proliferative activities of MC3T3-E1 cells in a concentration-dependent manner. Our data indicate that 5-HT fill the role for proliferation of osteoblast cells in their premature period. Notably, 5-HT{sub 2A} receptor may be functionally expressed to regulate mechanisms underlying osteoblast cell proliferation, at least in part, through activation of ERK/MAPK pathways in MC3T3-E1 cells.

  8. The effect of glucose concentration on insulin-induced 3T3-L1 adipose cell differentiation.

    PubMed

    Gagnon, A; Sorisky, A

    1998-03-01

    We examined the effect of glucose concentration on insulin-induced 3T3-L1 adipose cell differentiation. Oil Red O staining of neutral lipid, cellular triglyceride mass, and glycerol phosphate dehydrogenase (GPDH) activity, were greater in 3T3-L1 cells cultured at 5 mM vs. 25 mM glucose. GPDH activity was 2- to 4-fold higher at 5 mM vs. 25 mM glucose over a range of insulin concentrations (0.1 to 100 nM). Insulin-stimulated tyrosine phosphorylation of insulin receptor substrate-1 (IRS-1) was 1.7-fold greater, and insulin-stimulated phosphoinositide 3-kinase association with IRS-1 was 2.3-fold higher, at 5 mM vs. 25 mM glucose. These effects of glucose were not caused by alterations in IRS-1 mass or cell-surface insulin binding. In preadipose cells at 5 mM glucose, expression of the leukocyte antigen-related (LAR) protein tyrosine phosphatase (negative regulator of insulin signaling) was 63% of the level at 25 mM glucose. Our data demonstrate that glucose concentration affects insulin-induced 3T3-L1 adipose cell differentiation as well as differentiation-directed insulin signaling pathways. Alterations in LAR expression potentially may be involved in modulating these responses. PMID:9545023

  9. Carnosic Acid Inhibits Lipid Accumulation in 3T3-L1 Adipocytes Through Attenuation of Fatty Acid Desaturation

    PubMed Central

    Park, Mi-Young; Sung, Mi-Kyung

    2015-01-01

    Background: Excess body fat accumulation contributes to the development of metabolic disorders that can cause adverse health effects. Carnosic acid (CA), a major bioactive component of rosemary (Rosemarinus officinalis), has been suggested to possess anti-adipogenic properties. The present study was conducted to elucidate the mechanism underlying the anti-adipogenic effects of CA. Methods: 3T3-L1 pre-adipocytes were treated with CA (0.1, 1, and 10 μM) from day 0 to day 8 of differentiation. On day 8, biochemical markers of lipid accumulation and the degree of fatty acid desaturation were measured. Results: Oil Red O staining results, triglyceride (TG) accumulation, and glycerol 3-phosphate dehydrogenase activity suggested that CA significantly inhibited lipid accumulation in 3T3-L1 adipocytes. CA significantly decreased mRNA expression of peroxisome proliferator-activated receptor-γ, sterol regulatory element-binding protein 1, and CCAAT/enhancer binding protein-α in a dose-dependent manner. Moreover, it decreased the ratio of both C16:1/C16:0 and C18:1/C18:0, with reduced expression of stearoyl CoA desaturase 1 mRNA and protein. Conclusions: These results suggest that CA efficiently suppressed adipogenesis in 3T3-L1 adipocytes and its action, at least in part, is associated with the downregulation of adipogenesis-related genes and the fatty acid composition of TG accumulated in adipocytes. PMID:25853102

  10. MiR-25 suppresses 3T3-L1 adipogenesis by directly targeting KLF4 and C/EBPα.

    PubMed

    Liang, Wei-Cheng; Wang, Yan; Liang, Pu-Ping; Pan, Xu-Qing; Fu, Wei-Ming; Yeung, Venus Sai-Ying; Lu, Ying-Fei; Wan, David Chi-Cheong; Tsui, Stephen Kwok-Wing; Tsang, Suk-Ying; Ma, Wen-Bin; Zhang, Jin-Fang; Waye, Mary Miu-Yee

    2015-11-01

    In the past decade, miRNA emerges as a vital player in orchestrating gene regulation and maintaining cellular homeostasis. It is well documented that miRNA influences a variety of biological events, including embryogenesis, cell fate decision, and cellular differentiation. Adipogenesis is an organized process of cellular differentiation by which pre-adipocytes differentiate towards mature adipocytes. It has been shown that adipogenesis is tightly modulated by a number of transcription factors such as PPARγ, KLF4, and C/EBPα. However, the molecular mechanisms underlying the missing link between miRNA and adipogenesis-related transcription factors remain elusive. In this study, we unveiled that miR-25, a member of miR-106b-25 cluster, was remarkably downregulated during 3T3-L1 adipogenesis. Restored expression of miR-25 significantly impaired 3T3-L1 adipogenesis and downregulated the expression of serial adipogenesis-related genes. Further experiments presented that ectopic expression of miR-25 did not affect cell proliferation and cell cycle progression. Finally, KLF4 and C/EBPα, two key regulators of adipocyte differentiation, were experimentally identified as bona fide targets for miR-25. These data indicate that miR-25 is a novel negative regulator of adipocyte differentiation and it suppressed 3T3-L1 adipogenesis by targeting KLF4 and C/EBPα, which provides novel insights into the molecular mechanism of miRNA-mediated cellular differentiation. PMID:25923408

  11. Collagen-derived dipeptide prolyl-hydroxyproline promotes differentiation of MC3T3-E1 osteoblastic cells

    SciTech Connect

    Kimira, Yoshifumi; Ogura, Kana; Taniuchi, Yuri; Kataoka, Aya; Inoue, Naoki; Sugihara, Fumihito; Nakatani, Sachie; Shimizu, Jun; Wada, Masahiro; Mano, Hiroshi

    2014-10-24

    Highlights: • Pro-Hyp did not affect MC3T3-E1 cell proliferation and matrix mineralization. • Pro-Hyp significantly increased alkaline phosphatase activity. • Pro-Hyp significantly upregulated gene expression of Runx2, Osterix, and Col1α1. - Abstract: Prolyl-hydroxyproline (Pro-Hyp) is one of the major constituents of collagen-derived dipeptides. The objective of this study was to investigate the effects of Pro-Hyp on the proliferation and differentiation of MC3T3-E1 osteoblastic cells. Addition of Pro-Hyp did not affect MC3T3-E1 cell proliferation and matrix mineralization but alkaline phosphatase activity was significantly increased. Furthermore, cells treated with Pro-Hyp significantly upregulated gene expression of Runx2, Osterix, and Col1α1. These results indicate that Pro-Hyp promotes osteoblast differentiation. This study demonstrates for the first time that Pro-Hyp has a positive effect on osteoblast differentiation with upregulation of Runx2, Osterix, and Collα1 gene expression.

  12. Endogenous lectins from cultured cells: nuclear localization of carbohydrate-binding protein 35 in proliferating 3T3 fibroblasts.

    PubMed Central

    Moutsatsos, I K; Wade, M; Schindler, M; Wang, J L

    1987-01-01

    Proliferating 3T3 mouse fibroblasts contain higher levels of the lectin carbohydrate-binding protein 35 (CBP35) than do quiescent cultures of the same cells. An immunofluorescence study was carried out with a rabbit antiserum directed against CBP35 to map the cellular fluorescence distribution in a large population of cells under different growth conditions. This cytometric analysis showed that the lectin is predominantly localized in the nucleus of the proliferating cells. In quiescent 3T3 cultures, the majority of the cells lost their nuclear staining and underwent a general decrease in the overall fluorescence intensity. Stimulation of serum-starved quiescent 3T3 cells by the addition of serum resulted in an increase in the level of CBP35. The percentage of cells showing distinct punctate intranuclear staining reached a maximum at about the same time as the onset of the first S-phase of the cell cycle. All of these results suggest that CBP35 may be a protein whose presence in the nucleus, in discrete punctate distribution, is coordinated with the proliferation state of the cell. Images PMID:3306680

  13. Aculeatin, a coumarin derived from Toddalia asiatica (L.) Lam., enhances differentiation and lipolysis of 3T3-L1 adipocytes

    SciTech Connect

    Watanabe, Akio; Kato, Tsuyoshi; Ito, Yusuke; Yoshida, Izumi; Harada, Teppei; Mishima, Takashi; Fujita, Kazuhiro; Watai, Masatoshi; Nakagawa, Kiyotaka; Miyazawa, Teruo

    2014-10-31

    Highlights: • Aculeatin promoted adipocyte differentiation. • Aculeatin improved glucose uptake. • Aculeatin enhanced adipocyte lipolysis. - Abstract: Toddalia asiatica (L.) Lam. (T. asiatica) has been utilized traditionally for medicinal purposes such as the treatment of diabetes. Currently, the extract is considered to be a good source of anti-diabetic agents, but the active compounds have yet to be identified. In this study, we investigated the effects of fractionated T. asiatica extracts on the differentiation of 3T3-L1 preadipocytes and identified aculeatin as a potential active agent. When 3T3-L1 preadipocytes were treated with aculeatin isolated from T. asiatica in the presence of insulin, aculeatin increased cellular triglyceride levels and glycerol-3-phosphate dehydrogenase activity. This indicated that aculeatin could enhance the differentiation of preadipocytes into adipocytes. Further analyses using a DNA microarray and real-time quantitative reverse-transcription PCR showed an increase in the expression of peroxisome proliferator-activated receptor-γ target genes (Pparg, Ap2, Cd36, Glut4 and Adipoq) by aculeatin, suggesting that aculeatin enhances the differentiation of 3T3-L1 cells by modulating the expression of genes critical for adipogenesis. Interestingly, after treatment of differentiated adipocytes with aculeatin, glucose uptake and lipolysis were enhanced. Overall, our results suggested that aculeatin is an active compound in T. asiatica for enhancing both differentiation and lipolysis of adipocytes, which are useful for the treatment of lipid abnormalities as well as diabetes.

  14. Coronary Artery Anomalies and Variants: Technical Feasibility of Assessment with Coronary MR Angiography at 3 T1

    PubMed Central

    Gharib, Ahmed M.; Ho, Vincent B.; Rosing, Douglas R.; Herzka, Daniel A.; Stuber, Matthias; Arai, Andrew E.; Pettigrew, Roderic I.

    2008-01-01

    The purpose of this study was to prospectively use a whole-heart three-dimensional (3D) coronary magnetic resonance (MR) angiography technique specifically adapted for use at 3 T and a parallel imaging technique (sensitivity encoding) to evaluate coronary arterial anomalies and variants (CAAV). This HIPAA-compliant study was approved by the local institutional review board, and informed consent was obtained from all participants. Twenty-two participants (11 men, 11 women; age range, 18–62 years) were included. Ten participants were healthy volunteers, whereas 12 participants were patients suspected of having CAAV. Coronary MR angiography was performed with a 3-T MR imager. A 3D free-breathing navigator-gated and vector electrocardiographically–gated segmented k-space gradient-echo sequence with adiabatic T2 preparation pulse and parallel imaging (sensitivity encoding) was used. Whole-heart acquisitions (repetition time msec/echo time msec, 4/1.35; 20° flip angle; 1 × 1 × 2-mm acquired voxel size) lasted 10–12 minutes. Mean examination time was 41 minutes ± 14 (standard deviation). Findings included aneurysms, ectasia, arteriovenous fistulas, and anomalous origins. The 3D whole-heart acquisitions developed for use with 3 T are feasible for use in the assessment of CAAV. © RSNA, 2008 PMID:18372470

  15. MR images of mouse brain using clinical 3T MR scanner and 4CH-Mouse coil

    NASA Astrophysics Data System (ADS)

    Lim, Soo Mee; Park, Eun Mi; Lyoo, In Kyoon; Lee, Junghyun; Han, Bo Mi; Lee, Jeong Kyong; Lee, Su Bin

    2015-07-01

    Objectives: Although small-bore high-field magnets are useful for research in small rodent models,this technology, however, has not been easily accessible to most researchers. This current study, thus,tried to evaluate the usability of 4CH-Mouse coil (Philips Healthcare, Best, the Netherlands) forpreclinical investigations in clinical 3T MR scan environment. We evaluated the effects of ischemicpreconditioning (IP) in the mouse stroke model with clinical 3T MR scanner and 4CH-Mouse coil. Materials and Methods: Experiments were performed on male C57BL/6 mice that either received the IP or sham operation (control). Three different MR sequences including diffusion weighted images (DWI), T2-weighted images (T2WI), and fluid attenuated inversion recovery (FLAIR) were performed on the mouse brains following 24, 72 hours of middle cerebral artery occlusion (MCAO) and analyzed for infarct lesions. Results: The images showed that the IP-treated mouse brains had significantly smaller infarct volumes compared to the control group. Of the MR sequences employed, the T2WI showed the highest level of correlations with postmortem infarct volume measurements. Conclusions: The clinical 3T MR scanner turned out to have a solid potential as a practical tool for imaging small animal brains. MR sequences including DWI, T2WI, FLAIR were obtained with acceptable resolution and in a reasonable time constraint in evaluating a mouse stroke model brain.

  16. Cocoa tea (Camellia ptilophylla) water extract inhibits adipocyte differentiation in mouse 3T3-L1 preadipocytes

    PubMed Central

    Li, Kai Kai; Liu, Chuek Lun; Shiu, Hoi Ting; Wong, Hing Lok; Siu, Wing Sum; Zhang, Cheng; Han, Xiao Qiang; Ye, Chuang Xing; Leung, Ping Chung; Ko, Chun Hay

    2016-01-01

    Cocoa tea (Camellia ptilophylla) is a naturally decaffeinated tea plant. Previously we found that cocoa tea demonstrated a beneficial effect against high-fat diet induced obesity, hepatic steatosis, and hyperlipidemia in mice. The present study aimed to investigate the anti-adipogenic effect of cocoa tea in vitro using preadipocytes 3T3-L1. Adipogenic differentiation was confirmed by Oil Red O stain, qPCR and Western blot. Our results demonstrated that cocoa tea significantly inhibited triglyceride accumulation in mature adipocytes in a dose-dependent manner. Cocoa tea was shown to suppress the expressions of key adipogenic transcription factors, including peroxisome proliferator-activated receptor gamma (PPAR γ) and CCAAT/enhancer binding protein (C/EBP α). The tea extract was subsequently found to reduce the expressions of adipocyte-specific genes such as sterol regulatory element binding transcription factor 1c (SREBP-1c), fatty acid synthase (FAS), Acetyl-CoA carboxylase (ACC), fatty acid translocase (FAT) and stearoylcoenzyme A desaturase-1 (SCD-1). In addition, JNK, ERK and p38 phosphorylation were inhibited during cocoa tea inhibition of 3T3-L1 adipogenic differentiation. Taken together, this is the first study that demonstrates cocoa tea has the capacity to suppress adipogenesis in pre-adipocyte 3T3-L1 similar to traditional green tea PMID:26833256

  17. Strategies of NF-κB signaling modulation by ectromelia virus in BALB/3T3 murine fibroblasts.

    PubMed

    Struzik, Justyna; Szulc-Dąbrowska, Lidia; Winnicka, Anna; Niemiałtowski, Marek

    2015-10-01

    Nuclear factor κB (NF-κB) is a pleiotropic transcription factor that regulates the expression of immune response genes. NF-κB signaling can be disrupted by pathogens that prevent host immune response. In this work, we examined the influence of ectromelia (mousepox) virus (ECTV) on NF-κB signaling in murine BALB/3T3 fibroblasts. Activation of NF-κB via tumor necrosis factor (TNF) receptor 1 (TNFR1) in these cells induces proinflammatory cytokine secretion. We show that ECTV does not recruit NF-κB to viral factories or induce NF-κB nuclear translocation in BALB/3T3 cells. Additionally, ECTV counteracts TNF-α-induced p65 NF-κB nuclear translocation during the course of infection. Inhibition of TNF-α-induced p65 nuclear translocation was also observed in neighboring cells that underwent fusion with ECTV-infected cells. ECTV inhibits the key step of NF-κB activation, i.e. Ser32 phosphorylation and degradation of inhibitor κBα (IκBα) induced by TNF-α. We also observed that ECTV prevents TNF-α-induced Ser536 of p65 phosphorylation in BALB/3T3 cells. Studying TNFR1 signaling provides information about regulation of inflammatory response and cell survival. Unraveling poxviral immunomodulatory strategies may be helpful in drug target identification as well as in vaccine development. PMID:26232502

  18. 4-Hydroxyderricin, as a PPARγ Agonist, Promotes Adipogenesis, Adiponectin Secretion, and Glucose Uptake in 3T3-L1 Cells.

    PubMed

    Li, Yongjia; Goto, Tsuyoshi; Yamakuni, Kanae; Takahashi, Haruya; Takahashi, Nobuyuki; Jheng, Huei-Fen; Nomura, Wataru; Taniguchi, Masahiko; Baba, Kimiye; Murakami, Shigeru; Kawada, Teruo

    2016-07-01

    Adipocyte differentiation plays a pivotal role in maintaining the production of small-size adipocytes with insulin sensitivity, and impaired adipogenesis is implicated in insulin resistance. 4-Hydroxyderricin (4-HD), a phytochemical component of Angelica keiskei, possesses diverse biological properties such as anti-inflammatory, antidiabetic, and antitumor. In the present study, we investigated the effects of 4-HD on adipocyte differentiation. 4-HD promoted lipid accumulation in 3T3-L1 cells, upregulated both peroxisome proliferator-activated receptor (PPAR)-γ mRNA and protein expression, and acted as a ligand for PPARγ in the luciferase assay. Moreover, 4-HD increased the mRNA and protein expression levels of adiponectin. Additionally, it promoted insulin-dependent glucose uptake into 3T3-L1 adipocytes and increased Akt phosphorylation and glucose transporter (GLUT) 4 mRNA expression. In summary, these findings suggest that 4-HD, which promoted adipogenesis and insulin sensitivity in 3T3-L1 cells, might be a phytochemical with potent insulin-sensitizing effects. PMID:27098252

  19. Cocoa tea (Camellia ptilophylla) water extract inhibits adipocyte differentiation in mouse 3T3-L1 preadipocytes.

    PubMed

    Li, Kai Kai; Liu, Chuek Lun; Shiu, Hoi Ting; Wong, Hing Lok; Siu, Wing Sum; Zhang, Cheng; Han, Xiao Qiang; Ye, Chuang Xing; Leung, Ping Chung; Ko, Chun Hay

    2016-01-01

    Cocoa tea (Camellia ptilophylla) is a naturally decaffeinated tea plant. Previously we found that cocoa tea demonstrated a beneficial effect against high-fat diet induced obesity, hepatic steatosis, and hyperlipidemia in mice. The present study aimed to investigate the anti-adipogenic effect of cocoa tea in vitro using preadipocytes 3T3-L1. Adipogenic differentiation was confirmed by Oil Red O stain, qPCR and Western blot. Our results demonstrated that cocoa tea significantly inhibited triglyceride accumulation in mature adipocytes in a dose-dependent manner. Cocoa tea was shown to suppress the expressions of key adipogenic transcription factors, including peroxisome proliferator-activated receptor gamma (PPAR γ) and CCAAT/enhancer binding protein (C/EBP α). The tea extract was subsequently found to reduce the expressions of adipocyte-specific genes such as sterol regulatory element binding transcription factor 1c (SREBP-1c), fatty acid synthase (FAS), Acetyl-CoA carboxylase (ACC), fatty acid translocase (FAT) and stearoylcoenzyme A desaturase-1 (SCD-1). In addition, JNK, ERK and p38 phosphorylation were inhibited during cocoa tea inhibition of 3T3-L1 adipogenic differentiation. Taken together, this is the first study that demonstrates cocoa tea has the capacity to suppress adipogenesis in pre-adipocyte 3T3-L1 similar to traditional green tea. PMID:26833256

  20. Isoflavones in Chickpeas Inhibit Adipocyte Differentiation and Prevent Insulin Resistance in 3T3-L1 Cells.

    PubMed

    Gao, Yue; Yao, Yang; Zhu, Yinging; Ren, Guixing

    2015-11-11

    Diabetes mellitus is a metabolic disease characterized by hyperglycemia arising from defects in insulin secretion. This study investigated the effects of isoflavones in chickpea sprouts germinated in light (IGL) and isoflavones in chickpea seeds (ICS) on insulin resistance through their role in suppression of 3T3-L1 adipocyte differentiation. Results showed that IGL and ICS inhibit the differentiation of 3T3-L1 pre-adipocytes induced by differentiation medium in a dose-dependent manner, and the suppressive effect of IGL was stronger (p < 0.05) than that of ICS, evidenced by a decrease of Oil Red O staining and intracellular triacylglycerol content in the mature adipocytes. IGL and ICS also stimulated glucose uptake significantly (p < 0.05). Besides, IGL and ICS treatment caused a significant decrease in mRNA and protein expression levels of adipogenesis-related transcription factors peroxisome proliferator-activated receptor γ (PPARγ) and CCAAT-enhancer-binding protein α (C/EBPα). Furthermore, the mRNA and protein expression levels of adipocyte fatty acid-binding protein (ap2), lipoprotein lipase (LPL), uncoupling protein-2 (UCP-2), and glucose transporter 4 (Glut4) in 3T3-L1 cells were also markedly down-regulated (p < 0.05). PMID:26494490

  1. Soy pinitol acts partly as an insulin sensitizer or insulin mediator in 3T3-L1 preadipocytes

    PubMed Central

    Do, Gyeong-Min; Choi, Myung-Sook; Kim, Hye-Jin; Woo, Myung-Nam; Lee, Mi-Kyung

    2007-01-01

    The blood glucose-lowering property of pinitol is mediated via the insulin signaling pathway. This study was carried out to evaluate the effects of soy pinitol on adipogenesis in a 3T3-L1 cell line; 3T3-L1 preadipocytes were treated with pinitol (0–1 mM) together with insulin for 9 days. The regulation of lipid metabolism was assessed by oil-red-O staining of intracellular lipids and real-time PCR of adipogenesis-related factors. The inhibition of cell proliferation was estimated by MTT assay. Pinitol treatment did not inhibit lipid accumulation, nor did it affect expression of adipogenesis-related factors, including ADD1, aP2 and FAS, in a dose-dependent manner. Expression of adiponectin, GLUT4, IRS, C/EBPα and PPARγ mRNAs, however, increased in cells treated with 0.5 mM and/or 1 mM pinitol. Pinitol treatment did not affect the inhibition of cell growth and proliferation in a dose-dependent manner. Accordingly, we suggest that pinitol is nontoxic to this cell line, and that it enhances adipogenesis by acting as an insulin sensitizer or insulin mediator via the upregulation of adiponectin, GLUT4, IRS, C/EBPα and PPARγ in 3T3-L1 preadipocytes. PMID:18850231

  2. Nickel-Refining Fumes Induced DNA Damage and Apoptosis of NIH/3T3 Cells via Oxidative Stress

    PubMed Central

    Wang, Yue; Wang, Sheng-Yuan; Jia, Li; Zhang, Lin; Ba, Jing-Chong; Han, Dan; Yu, Cui-Ping; Wu, Yong-Hui

    2016-01-01

    Although there have been numerous studies examining the toxicity and carcinogenicity of nickel compounds in humans and animals, its molecular mechanisms of action are not fully elucidated. In our research, NIH/3T3 cells were exposed to nickel-refining fumes at the concentrations of 0, 6.25, 12.50, 25, 50 and 100 μg/mL for 24 h. Cell viability, cell apoptosis, reactive oxygen species (ROS) level, lactate dehydrogenase (LDH) assay, the level of glutathione (GSH), activities of superoxide dismutase (SOD), catalase (CAT), and malondialdehyde (MDA) level were detected. The exposure of NIH/3T3 cells to nickel-refining fumes significantly reduced cell viability and induced cell apoptotic death in a dose-dependent manner. Nickel-refining fumes significantly increased ROS levels and induced DNA damage. Nickel-refining fumes may induce the changes in the state of ROS, which may eventually initiate oxidative stress, DNA damage and apoptosis of NIH/3T3 cells. PMID:27347984

  3. Modest hypoxia significantly reduces triglyceride content and lipid droplet size in 3T3-L1 adipocytes

    SciTech Connect

    Hashimoto, Takeshi; Yokokawa, Takumi; Endo, Yuriko; Iwanaka, Nobumasa; Higashida, Kazuhiko; Taguchi, Sadayoshi

    2013-10-11

    Highlights: •Long-term hypoxia decreased the size of LDs and lipid storage in 3T3-L1 adipocytes. •Long-term hypoxia increased basal lipolysis in 3T3-L1 adipocytes. •Hypoxia decreased lipid-associated proteins in 3T3-L1 adipocytes. •Hypoxia decreased basal glucose uptake and lipogenic proteins in 3T3-L1 adipocytes. •Hypoxia-mediated lipogenesis may be an attractive therapeutic target against obesity. -- Abstract: Background: A previous study has demonstrated that endurance training under hypoxia results in a greater reduction in body fat mass compared to exercise under normoxia. However, the cellular and molecular mechanisms that underlie this hypoxia-mediated reduction in fat mass remain uncertain. Here, we examine the effects of modest hypoxia on adipocyte function. Methods: Differentiated 3T3-L1 adipocytes were incubated at 5% O{sub 2} for 1 week (long-term hypoxia, HL) or one day (short-term hypoxia, HS) and compared with a normoxia control (NC). Results: HL, but not HS, resulted in a significant reduction in lipid droplet size and triglyceride content (by 50%) compared to NC (p < 0.01). As estimated by glycerol release, isoproterenol-induced lipolysis was significantly lowered by hypoxia, whereas the release of free fatty acids under the basal condition was prominently enhanced with HL compared to NC or HS (p < 0.01). Lipolysis-associated proteins, such as perilipin 1 and hormone-sensitive lipase, were unchanged, whereas adipose triglyceride lipase and its activator protein CGI-58 were decreased with HL in comparison to NC. Interestingly, such lipogenic proteins as fatty acid synthase, lipin-1, and peroxisome proliferator-activated receptor gamma were decreased. Furthermore, the uptake of glucose, the major precursor of 3-glycerol phosphate for triglyceride synthesis, was significantly reduced in HL compared to NC or HS (p < 0.01). Conclusion: We conclude that hypoxia has a direct impact on reducing the triglyceride content and lipid droplet size via

  4. SU-E-T-508: End to End Testing of a Prototype Eclipse Module for Planning Modulated Arc Therapy On the Siemens Platform

    SciTech Connect

    Huang, L; Sarkar, V; Spiessens, S; Rassiah-Szegedi, P; Huang, Y; Salter, B; Zhao, H; Szegedi, M

    2014-06-01

    Purpose: The latest clinical implementation of the Siemens Artiste linac allows for delivery of modulated arcs (mARC) using full-field flattening filter free (FFF) photon beams. The maximum doserate of 2000 MU/min is well suited for high dose treatments such as SBRT. We tested and report on the performance of a prototype Eclipse TPS module supporting mARC capability on the Artiste platform. Method: our spine SBRT patients originally treated with 12/13 field static-gantry IMRT (SGIMRT) were chosen for this study. These plans were designed to satisfy RTOG0631 guidelines with a prescription of 16Gy in a single fraction. The cases were re-planned as mARC plans in the prototype Eclipse module using the 7MV FFF beam and required to satisfy RTOG0631 requirements. All plans were transferred from Eclipse, delivered on a Siemens Artiste linac and dose-validated using the Delta4 system. Results: All treatment plans were straightforwardly developed, in timely fashion, without challenge or inefficiency using the prototype module. Due to the limited number of segments in a single arc, mARC plans required 2-3 full arcs to yield plan quality comparable to SGIMRT plans containing over 250 total segments. The average (3%/3mm) gamma pass-rate for all arcs was 98.5±1.1%, thus demonstrating both excellent dose prediction by the AAA dose algorithm and excellent delivery fidelity. Mean delivery times for the mARC plans(10.5±1.7min) were 50-70% lower than the SGIMRT plans(26±2min), with both delivered at 2000 MU/min. Conclusion: A prototype Eclipse module capable of planning for Burst Mode modulated arc delivery on the Artiste platform has been tested and found to perform efficiently and accurately for treatment plan development and delivered-dose prediction. Further investigation of more treatment sites is being carried out and data will be presented.

  5. Expedition 34 Welcomes New Trio

    NASA Video Gallery

    The Soyuz TMA-07M carrying new Expedition 34 crew members Chris Hadfield, Roman ROmanenko and Tom Marshburn docked to the International Space Station’s Rassvet module at 9:09 a.m. EST on Friday. ...

  6. A Trio of Brownian Donkeys

    NASA Astrophysics Data System (ADS)

    van den Broeck, C.; Cleuren, B.; Kawai, R.; Kambon, M.

    A previously introduced model (B. Cleuren and C. Van den Broeck, Europhys. Lett. 54, 1 (2001)) is studied numerically. Pure negative mobility is found for the minimum number of three interacting walkers.

  7. 26 CFR 1.1092(b)-3T - Mixed straddles; straddle-by-straddle identification under section 1092(b)(2)(A)(i)(I) (temporary).

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... identification under section 1092(b)(2)(A)(i)(I) (temporary). 1.1092(b)-3T Section 1.1092(b)-3T Internal Revenue... section 1092(b)(2)(A)(i)(I) (temporary). (a) In general. Except as otherwise provided, a taxpayer shall... constitute independent verification: (i) Separate account. Placement of one or more positions of a......

  8. 26 CFR 1.905-3T - Adjustments to United States tax liability and to the pools of post-1986 undistributed earnings...

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 26 Internal Revenue 9 2010-04-01 2010-04-01 false Adjustments to United States tax liability and to the pools of post-1986 undistributed earnings and post-1986 foreign income taxes as a result of a foreign tax redetermination (temporary). 1.905-3T Section 1.905-3T Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY...

  9. 26 CFR 1.905-3T - Adjustments to United States tax liability and to the pools of post-1986 undistributed earnings...

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 26 Internal Revenue 9 2014-04-01 2014-04-01 false Adjustments to United States tax liability and to the pools of post-1986 undistributed earnings and post-1986 foreign income taxes as a result of a foreign tax redetermination (temporary). 1.905-3T Section 1.905-3T Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY...

  10. 26 CFR 1.905-3T - Adjustments to United States tax liability and to the pools of post-1986 undistributed earnings...

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 26 Internal Revenue 9 2012-04-01 2012-04-01 false Adjustments to United States tax liability and to the pools of post-1986 undistributed earnings and post-1986 foreign income taxes as a result of a foreign tax redetermination (temporary). 1.905-3T Section 1.905-3T Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY...

  11. 26 CFR 1.905-3T - Adjustments to United States tax liability and to the pools of post-1986 undistributed earnings...

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 26 Internal Revenue 9 2013-04-01 2013-04-01 false Adjustments to United States tax liability and to the pools of post-1986 undistributed earnings and post-1986 foreign income taxes as a result of a foreign tax redetermination (temporary). 1.905-3T Section 1.905-3T Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY...

  12. 26 CFR 1.905-3T - Adjustments to United States tax liability and to the pools of post-1986 undistributed earnings...

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 26 Internal Revenue 9 2011-04-01 2011-04-01 false Adjustments to United States tax liability and to the pools of post-1986 undistributed earnings and post-1986 foreign income taxes as a result of a foreign tax redetermination (temporary). 1.905-3T Section 1.905-3T Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY...

  13. Signal-to-noise ratio increase in carotid atheroma MRI: a comparison of 1.5 and 3 T

    PubMed Central

    Young, V E; Patterson, A J; Tunnicliffe, E M; Sadat, U; Graves, M J; Tang, T Y; Priest, A N; Kirkpatrick, P J; Gillard, J H

    2012-01-01

    Objectives This study reports quantitative comparisons of signal-to-noise ratio (SNR) at 1.5 and 3 T from images of carotid atheroma obtained using a multicontrast, cardiac-gated, blood-suppressed fast spin echo protocol. Methods 18 subjects, with carotid atherosclerosis (>30% stenosis) confirmed on ultrasound, were imaged on both 1.5 and 3 T systems using phased-array coils with matched hardware specifications. T1 weighted (T1W), T2 weighted (T2W) and proton density-weighted (PDW) images were acquired with identical scan times. Multiple slices were prescribed to encompass both the carotid bifurcation and the plaque. Image quality was quantified using the SNR and contrast-to-noise ratio (CNR). A phantom experiment was also performed to validate the SNR method and confirm the size of the improvement in SNR. Comparisons of the SNR values from the vessel wall with muscle and plaque/lumen CNR measurements were performed at a patient level. To account for the multiple comparisons a Bonferroni correction was applied. Results One subject was excluded from the protocol owing to image quality and protocol failure. The mean improvement in SNR in plaque was 1.9, 2.1 and 2.1 in T1W, T2W and PDW images, respectively. All plaque SNR improvements were statistically significant at the p<0.05 level. The phantom experiment reported an improvement in SNR of 2.4 for PDW images. Conclusions Significant gains in SNR can be obtained for carotid atheroma imaging at 3 T compared with 1.5 T. There was also a trend towards increased CNR. However, this was not significant after the application of the Bonferroni correction. PMID:22294703

  14. Widdrol-induced lipolysis is mediated by PKC and MEK/ERK in 3T3-L1 adipocytes.

    PubMed

    Jeong, Hyun Young; Yun, Hee Jung; Kim, Byung Woo; Lee, Eun Woo; Kwon, Hyun Ju

    2015-12-01

    Obesity is a serious medical condition causing various diseases such as heart disease, type-2 diabetes, and cancer. Fat cells (adipocytes) play an important role in the generation of energy through hydrolysis of lipids they accumulate. Therefore, induction of lipolysis (breakdown of lipids into fatty acids and glycerol), is one of the ways to treat obesity. In the present study, we investigated the lipolytic effect of widdrol in 3T3-L1 adipocytes and its mechanism. Widdrol considerably increased the amount of glycerol released from 3T3-L1 adipocytes into the medium in a time- and dose-dependent manner. To determine the mechanism of this effect, we investigated the alterations in glycerol release and protein expression in 3T3-L1 adipocytes treated with widdrol alone or widdrol and inhibitors of proteins involved in the cAMP-dependent pathway or cAMP-independent PKC-MAPK pathway, which are known to induce lipolysis in adipocytes. The adenylyl cyclase inhibitor SQ-22536, PLA2 inhibitor dexamethasone, PI3K inhibitor wortmannin, and PKA inhibitor H-89, which were used to investigate the involvement of the cAMP-dependent pathway, did not affect the lipolytic effect of widdrol. Widdrol-induced phosphorylation of PKC, MEK, and ERK, which are related to the PKC-MAPK pathway, and their phosphorylation was inhibited by their inhibitors (H-7, U0126, and PD-98059, respectively). Moreover, the increase in glycerol release induced by widdrol was almost completely blocked by PKC, MEK, and ERK inhibitors. These results suggest that widdrol induces lipolysis through activation of the PKC-MEK-ERK pathway. PMID:26359088

  15. Effect of botulinum neurotoxin type A (BoNTA) on the morphology and viability of 3T3 murine fibroblasts

    PubMed Central

    Bandala, Cindy; Terán-Melo, Juan Luis; Anaya-Ruiz, Maricruz; Mejía-Barradas, Cesar Miguel; Domínguez-Rubio, Rene; la Garza-Montano, Paloma De; Alfaro-Rodríguez, Alfonso; Lara-Padilla, Eleazar

    2015-01-01

    Aim: BoNTA is used in the treatment of ophthalmological disorders, muscular hyperactivity and pain. In recent years it has been described that BoNTA reduces cellular viability and induces apoptosis in prostate cells lines. Studies about the effect of BoNTA are no well known. There have been studies about the effect of BoNTA on the expression levels of collagenase in fibroblasts, but not on its morphological impact on these cells. The aim of this study was to determine the effect of BoNTA on the morphology and viability of the 3T3 fibroblast cell line. Material and methods: The 3T3 fibroblast cell line was cultured and the experimental group received 10 U BoNTA added to a 0.9% sterile saline solution in a reconstituted vial. The control group received saline solution only. Cultured cells were observed and photographed at 5, 10, 15 and 20 h. Cell viability was evaluated by means of the trypan blue test, and cell proliferation with the Proliferation Assay kit (PROMEGA). Results: The application of BoNTA to 3T3 fibroblast cells induced morphological changes, such as a loss of normal fibroblast morphology. Additionally, we observed the cytoplasmic retraction and spread phenomena. The nuclei showed other important changes with Giemsa staining. Conclusion: The results indicate that BoNTA induced a loss of spindle form, increase in cytoplasmic vesicles, and the presence of nuclear vesicles (compacted chromatin surrounded by a nuclear envelope). This suggests an apoptotic process and decreased cell viability. Further studies are needed to explore the mechanisms of these alterations. PMID:26464704

  16. Radicicol, a heat shock protein 90 inhibitor, inhibits differentiation and adipogenesis in 3T3-L1 preadipocytes

    SciTech Connect

    He, Yonghan; Li, Ying; Zhang, Shuocheng; Perry, Ben; Zhao, Tiantian; Wang, Yanwen; Sun, Changhao

    2013-06-28

    Highlights: •Radicicol suppressed intracellular fat accumulation in 3T3-L1 adipocytes. •Radicicol inhibited the expression of FAS and FABP4. •Radicicol blocked cell cycle at the G1-S phase during cell differentiation. •Radicicol inhibited the PDK1/Akt pathway in adipocyte differentiation. -- Abstract: Heat shock protein 90 (Hsp90) is involved in various cellular processes, such as cell proliferation, differentiation and apoptosis. As adipocyte differentiation plays a critical role in obesity development, the present study investigated the effect of an Hsp90 inhibitor radicicol on the differentiation of 3T3-L1 preadipocytes and potential mechanisms. The cells were treated with different concentrations of radicicol during the first 8 days of cell differentiation. Adipogenesis, the expression of adipogenic transcriptional factors, differentiation makers and cell cycle were determined. It was found that radicicol dose-dependently decreased intracellular fat accumulation through down-regulating the expression of peroxisome proliferator-activated receptor γ (PPAR{sub γ}) and CCAAT element binding protein α (C/EBP{sub α}), fatty acid synthase (FAS) and fatty acid-binding protein 4 (FABP4). Flow cytometry analysis revealed that radicicol blocked cell cycle at G1-S phase. Radicicol redcued the phosphorylation of Akt while showing no effect on β-catenin expression. Radicicol decreased the phosphorylation of phosphoinositide-dependent kinase 1 (PDK1). The results suggest that radicicol inhibited 3T3-L1 preadipocyte differentiation through affecting the PDK1/Akt pathway and subsequent inhibition of mitotic clonal expansion and the expression/activity of adipogenic transcriptional factors and their downstream adipogenic proteins.

  17. Repeatability and sensitivity of T2* measurements in patients with head and neck squamous cell carcinoma at 3T

    PubMed Central

    Panek, Rafal; Welsh, Liam; Dunlop, Alex; Wong, Kee H.; Riddell, Angela M.; Koh, Dow‐Mu; Schmidt, Maria A.; Doran, Simon; Mcquaid, Dualta; Hopkinson, Georgina; Richardson, Cheryl; Nutting, Christopher M.; Bhide, Shreerang A.; Harrington, Kevin J.; Robinson, Simon P.; Newbold, Kate L.

    2016-01-01

    Purpose To determine whether quantitation of T2* is sufficiently repeatable and sensitive to detect clinically relevant oxygenation levels in head and neck squamous cell carcinoma (HNSCC) at 3T. Materials and Methods Ten patients with newly diagnosed locally advanced HNSCC underwent two magnetic resonance imaging (MRI) scans between 24 and 168 hours apart prior to chemoradiotherapy treatment. A multiple gradient echo sequence was used to calculate T2* maps. A quadratic function was used to model the blood transverse relaxation rate as a function of blood oxygenation. A set of published coefficients measured at 3T were incorporated to account for tissue hematocrit levels and used to plot the dependence of fractional blood oxygenation (Y) on T2* values, together with the corresponding repeatability range. Repeatability of T2* using Bland–Altman analysis, and calculation of limits of agreement (LoA), was used to assess the sensitivity, defined as the minimum difference in fractional blood oxygenation that can be confidently detected. Results T2* LoA for 22 outlined tumor volumes were 13%. The T2* dependence of fractional blood oxygenation increases monotonically, resulting in increasing sensitivity of the method with increasing blood oxygenation. For fractional blood oxygenation values above 0.11, changes in T2* were sufficient to detect differences in blood oxygenation greater than 10% (Δ T2* > LoA for ΔY > 0.1). Conclusion Quantitation of T2* at 3T can detect clinically relevant changes in tumor oxygenation within a wide range of blood volumes and oxygen tensions, including levels reported in HNSCC. J. Magn. Reson. Imaging 2016;44:72–80. PMID:26800280

  18. Growth hormone promoted tyrosyl phosphorylation of growth hormone receptors in murine 3T3-F442A fibroblasts and adipocytes

    SciTech Connect

    Foster, C.M.; Shafer, J.A.; Rozsa, F.W.; Wang, X.; Lewis, S.D.; Renken, D.A.; Natale, J.E.; Schwartz, J.; Carter-Su, C.

    1988-01-12

    Because many growth factor receptors are ligand-activated tyrosine protein kinases, the possibility that growth hormone (GH), a hormone implicated in human growth, promotes tyrosyl phosphorylation of its receptor was investigated. /sup 125/I-Labeled human GH was covalently cross-linked to receptors in intact 3T3-F442A fibroblasts, a cell line which differentiates into adipocytes in response to GH. The cross-linked cells were solubilized and passed over a column of phosphotyrosyl binding antibody immobilized on protein A-Sepharose. Immunoadsorbed proteins were eluted with a hapten (p-nitrophenyl phosphate) and analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autoradiography. The eluate from the antibody column contained in M/sub r/ 134,000 /sup 125/I-GH-receptor complex. A similar result was obtained when the adipocyte form of 3T3-F442A cells was used in place of fibroblast form. O-Phosphotyrosine prevented /sup 125/I-GH-receptor complexes from binding to the antibody column, whereas O-phosphoserine and O-phosphothreonine did not. In studies of GH-promoted phosphorylation in 3T3-F442A fibroblasts labeled metabolically with (/sup 32/P)P/sub i/, GH was shown to stimulate formation of a /sup 32/P-labeled protein which bound to immobilized phosphotyrosyl binding antibodies. The molecular weight of 114,000 obtained for this protein is similar to that expected for non-cross-linked GH receptor. These observations provide strong evidence that binding of GH to its receptor stimulates phosphorylation of tyrosyl residues in the GH receptor.

  19. TNF-α Induces Caspase-1 Activation Independently of Simultaneously Induced NLRP3 in 3T3-L1 Cells.

    PubMed

    Furuoka, Mana; Ozaki, Kei-Ichi; Sadatomi, Daichi; Mamiya, Sayaka; Yonezawa, Tomo; Tanimura, Susumu; Takeda, Kohsuke

    2016-12-01

    The intracellular cysteine protease caspase-1 is critically involved in obesity-induced inflammation in adipose tissue. A substantial body of evidence from immune cells, such as macrophages, has shown that caspase-1 activation depends largely on a protein complex, called the NLRP3 inflammasome, which consists of the NOD-like receptor (NLR) family protein NLRP3, the adaptor protein ASC, and caspase-1 itself. However, it is not fully understood how caspase-1 activation is regulated within adipocytes upon inflammatory stimuli. In this study, we show that TNF-α-induced activation of caspase-1 is accompanied by robust induction of NLRP3 in 3T3-L1 adipocytes but that caspase-1 activation may not depend on the NLRP3 inflammasome. Treatment of 3T3-L1 cells with TNF-α induced mRNA expression and activation of caspase-1. Although the basal expression of NLRP3 and ASC was undetectable in unstimulated cells, TNF-α strongly induced NLRP3 expression but did not induce ASC expression. Interestingly, inhibitors of the ERK MAP kinase pathway strongly suppressed NLRP3 expression but did not suppress the expression and activation of caspase-1 induced by TNF-α, suggesting that NLRP3 is dispensable for TNF-α-induced caspase-1 activation. Moreover, we did not detect the basal and TNF-α-induced expression of other NLR proteins (NLRP1a, NLRP1b, and NLRC4), which do not necessarily require ASC for caspase-1 activation. These results suggest that TNF-α induces caspase-1 activation in an inflammasome-independent manner in 3T3-L1 cells and that the ERK-dependent expression of NLRP3 may play a role independently of its canonical role as a component of inflammasomes. J. Cell. Physiol. 231: 2761-2767, 2016. © 2016 Wiley Periodicals, Inc. PMID:26989816

  20. Pasteurella multocida toxin (PMT) upregulates CTGF which leads to mTORC1 activation in Swiss 3T3 cells.

    PubMed

    Oubrahim, Hammou; Wong, Allison; Wilson, Brenda A; Chock, P Boon

    2013-05-01

    Pasteurella multocida toxin (PMT) is a mitogenic protein that hijacks cellular signal transduction pathways via deamidation of heterotrimeric G proteins. We previously showed that rPMT activates mTOR signaling via a Gαq/11/PLCβ/PKC mediated pathway, leading in part to cell proliferation and migration. Herein, we show that mTOR and MAPK, but not membrane-associated tyrosine kinases, are activated in serum-starved 3T3 cells by an autocrine/paracrine substance(s) secreted into the conditioned medium following rPMT treatment. Surprisingly, this diffusible factor(s) is capable of activating mTOR and MAPK pathways even in MEF Gαq/11 double knockout cells. Microarray analysis identified connective tissue growth factor (CTGF) mRNA as the most upregulated gene in rPMT-treated serum-starved 3T3 cells relative to untreated cells. These results were further confirmed using RT-PCR and Western blot analyses. In accord with rPMT-induced mTOR activation, upregulation of CTGF protein was observed in WT MEF, but not in Gαq/11 double knockout MEF cells. Although CTGF expression is regulated by TGFβ, rPMT did not activate TGFβ pathway. In addition, MEK inhibitors U0126 or PD98059, but not mTOR specific inhibitors, rapamycin and Torin 1, inhibited rPMT-induced upregulation of CTGF. Importantly, CTGF overexpression in serum-starved 3T3 cells using adenovirus led to phosphorylation of ribosomal protein S6, a downstream target of mTOR. However, despite the ability of CTGF to activate the mTOR pathway, upregulation of CTGF alone could not induce morphological changes as those observed in rPMT-treated cells. Our findings reveal that CTGF plays an important role, but there are additional factors involved in the mitogenic action of PMT. PMID:23415771

  1. A distinct cation-sensing mechanism in MC3T3-E1 osteoblasts functionally related to the calcium receptor.

    PubMed

    Quarles, L D; Hartle, J E; Siddhanti, S R; Guo, R; Hinson, T K

    1997-03-01

    The presence of a cation-sensing mechanism in osteoblasts is suggested by the ability of specific cations to stimulate osteoblastic proliferation in culture and to induce de novo bone formation in some experimental models. Our study examines whether extracellular cations stimulate osteoblasts through the recently identified G protein-coupled calcium receptor (CaR). We found that CaR agonists, calcium (Ca2+), gadolinium (Gd3+), aluminum (Al3+), and neomycin, stimulated DNA synthesis in murine-derived MC3T3-E1 preosteoblasts, whereas magnesium (Mg2+), nickel (Ni2+), cadmium (Cd2+), and zinc (Zn2+) had no effect. With the exception of Mg2+, the cation specificities and apparent affinities were similar to that reported for CaR. CaR agonists also stimulated DNA synthesis in C3HT10(1/2) fibroblasts, but not in mesangial PVG, CHO, hepatic HTC, COS-7 cells, or malignant transformed ROS17/2.8 and UMR-106 osteoblasts. In addition, similar to other growth factors, CaR agonists activated transcription of a serum response element luciferase reporter construct (SRE-Luc) stably transfected into MC3T3-E1 osteoblasts, but had no effect on SRE-Luc transfected into CHO and COS-7 cells. We were unable to detect CaR expression by Northern analysis using a mouse CaR-specific probe or to amplify CaR mRNA by reverse transcribed polymerase chain reaction in MC3T3-E1 osteoblasts. These findings suggest that an extra-cellular cation-sensing mechanism is present in murine-derived osteoblasts that is functionally similar to but molecularly distinct from CaR. PMID:9076582

  2. Nano-hydroxyapatite particles induce apoptosis on MC3T3-E1 cells and tissue cells in SD rats

    NASA Astrophysics Data System (ADS)

    Wang, Liting; Zhou, Gang; Liu, Haifeng; Niu, Xufeng; Han, Jingyun; Zheng, Lisha; Fan, Yubo

    2012-04-01

    While the advantages of nanomaterials are being increasingly recognized, their potential toxicity is drawing more and more attention and concern. In this study, we explore the toxicity mechanism of 20-30 nm rod-shaped hydroxyapatite (HA) nanoparticles in vitro and in vivo. The nanoparticles were prepared by precipitation and characterized by IR, XRD and TEM. Concentrations of 0 μg mL-1, 10 μg mL-1, 100 μg mL-1, 1 mg mL-1, and 10 mg mL-1 were applied to the MC3T3-E1 cells for viability (MTT-test). Based on the characteristic differences of the two methods of cell death, the morphological features of the MC3T3-E1 cell line co-cultured with nano-hydroxyapatite (n-HA) (10 mg mL-1) for 24 h were also observed by TEM. Furthermore, important serum biochemical markers and histopathological examinations were used to evaluate the potential toxicological effect of n-HA on the major organs of SD rats injected intraperitoneally with n-HA (33.3 mg kg-1 body weight). In the results, we found cell growth inhibition and apoptosis in MC3T3-E1 cells co-cultured with n-HA. Moreover, apoptosis but not necrosis was illustrated in liver and renal tissue by using histopathology slices and serum biochemical markers. It suggests that apoptosis may be the possible mechanism of n-HA toxicity and provides a better understanding of the biocompatibility of nanomaterials applied in human bone repair.

  3. Multi-contrast T2(⁎)-relaxometry upon visual stimulation at 3T and 7T.

    PubMed

    Berger, Moritz C; Bachert, Peter; Gröbner, Jens; Nagel, Armin M

    2016-09-01

    This study aims to quantify the mean change of the effective transverse relaxation time T2(⁎) in active brain regions of human volunteers at field strengths of B0=3T and 7T. Besides the mono-exponential signal decay model an extended model is tested that considers mesoscopic field gradients across imaging voxels. Both models are checked for cross-talk and correlations between the parameters. A visual checkerboard-stimulation experiment with pause and stimulation periods of 50s and six repetitions was performed on healthy volunteers. Eleven contrasts were acquired in about 1.47s/1.43s at 3T/7T using a segmented multi-contrast echo-planar imaging (EPI) sequence. Average BOLD-signal time courses were calculated in a multi-step (non-)linear least-squares process. Baseline T2(⁎) values of 37.72ms/24.99ms (47.34ms/33.71ms) with stimulus-correlated changes ∆T2(⁎)of 1.32ms/0.74ms (1.99ms/1.43ms) resulted from the mono-exponential (extended) model for 3T/7T. A dependence of those values on the initial intensity S0 was observed. Stimulus-correlated changes of S0 in the order of 1% were measured at both field strengths. The mono-exponential model was found to be less prone to instabilities in the regression of both parameters. Signal alterations due to inflow were observed. Measured relaxation times agree with values from literature using repetitive stimulation. A strong dependence of the measured relaxation times on the inflow-related model parameter was found for both models. The extended model is applicable to dynamic neurofunctional measurements, but is currently limited due to the low number of contrasts acquired. PMID:27046057

  4. 12/15-lipoxygenase products induce inflammation and impair insulin signaling in 3T3-L1 adipocytes.

    PubMed

    Chakrabarti, Swarup K; Cole, Banumathi K; Wen, Yeshao; Keller, Susanna R; Nadler, Jerry L

    2009-09-01

    Inflammation and insulin resistance associated with visceral obesity are important risk factors for the development of type 2 diabetes, atherosclerosis, and the metabolic syndrome. The 12/15-lipoxygenase (12/15-LO) enzyme has been linked to inflammatory changes in blood vessels that precede the development of atherosclerosis. The expression and role of 12/15-LO in adipocytes have not been evaluated. We found that 12/15-LO mRNA was dramatically upregulated in white epididymal adipocytes of high-fat fed mice. 12/15-LO was poorly expressed in 3T3-L1 fibroblasts and was upregulated during differentiation into adipocytes. Interestingly, the saturated fatty acid palmitate, a major component of high fat diets, augmented expression of 12/15-LO in vitro. When 3T3-L1 adipocytes were treated with the 12/15-LO products, 12-hydroxyeicosatetranoic acid (12(S)-HETE) and 12-hydroperoxyeicosatetraenoic acid (12(S)-HPETE), expression of proinflammatory cytokine genes, including tumor necrosis factor-alpha (TNF-alpha), monocyte chemoattractant protein 1 (MCP-1), interleukin 6 (IL-6), and IL-12p40, was upregulated whereas anti-inflammatory adiponectin gene expression was downregulated. 12/15-LO products also augmented c-Jun N-terminal kinase 1 (JNK-1) phosphorylation, a known negative regulator of insulin signaling. Consistent with impaired insulin signaling, we found that insulin-stimulated 3T3-L1 adipocytes exhibited decreased IRS-1(Tyr) phosphorylation, increased IRS-1(Ser) phosphorylation, and impaired Akt phosphorylation when treated with 12/15-LO product. Taken together, our data suggest that 12/15-LO products create a proinflammatory state and impair insulin signaling in 3T3-L1 adipocytes. Because 12/15-LO expression is upregulated in visceral adipocytes by high-fat feeding in vivo and also by addition of palmitic acid in vitro, we propose that 12/15-LO plays a role in promoting inflammation and insulin resistance associated with obesity. PMID:19521344

  5. Bioconversion of Citrus unshiu peel extracts with cytolase suppresses adipogenic activity in 3T3-L1 cells

    PubMed Central

    Lim, Heejin; Yeo, Eunju; Song, Eunju; Chang, Yun-Hee; Han, Bok-Kyung; Choi, Hyuk-Joon

    2015-01-01

    BACKGROUND/OBJECTIVES Citrus flavonoids have a variety of physiological properties such as anti-oxidant, anti-inflammation, anti-cancer, and anti-obesity. We investigated whether bioconversion of Citrus unshiu with cytolase (CU-C) ameliorates the anti-adipogenic effects by modulation of adipocyte differentiation and lipid metabolism in 3T3-L1 cells. MATERIALS/METHODS Glycoside forms of Citrus unshiu (CU) were converted into aglycoside forms with cytolase treatment. Cell viability of CU and CU-C was measured at various concentrations in 3T3L-1 cells. The anti-adipogenic and lipolytic effects were examined using Oil red O staining and free glycerol assay, respectively. We performed real time-polymerase chain reaction and western immunoblotting assay to detect mRNA and protein expression of adipogenic transcription factors, respectively. RESULTS Treatment with cytolase decreased flavanone rutinoside forms (narirutin and hesperidin) and instead, increased flavanone aglycoside forms (naringenin and hesperetin). During adipocyte differentiation, 3T3-L1 cells were treated with CU or CU-C at a dose of 0.5 mg/ml. Adipocyte differentiation was inhibited in CU-C group, but not in CU group. CU-C markedly suppressed the insulin-induced protein expression of CCAAT/enhancer-binding protein α (C/EBPα) and peroxisome proliferator-activated receptor gamma (PPARγ) as well as the mRNA levels of CEBPα, PPARγ, and sterol regulatory element binding protein 1c (SREBP1c). Both CU and CU-C groups significantly increased the adipolytic activity with the higher release of free glycerol than those of control group in differentiated 3T3-L1 adipocytes. CU-C is particularly superior in suppression of adipogenesis, whereas CU-C has similar effect to CU on stimulation of lipolysis. CONCLUSIONS These results suggest that bioconversion of Citrus unshiu peel extracts with cytolase enhances aglycoside flavonoids and improves the anti-adipogenic metabolism via both inhibition of key adipogenic

  6. True Pathologic Abnormality versus Artifact Foot Position and Magic Angle Artifact in the Peroneal Tendons with 3T Imaging.

    PubMed

    Horn, Deena B; Meyers, Steven; Astor, William

    2015-09-01

    Magnetic resonance imaging is a commonly ordered examination by many foot and ankle surgeons for ankle pain and suspected peroneal tendon pathologic abnormalities. Magic angle artifact is one of the complexities associated with this imaging modality. Magic angle refers to the increased signal on magnetic resonance images associated with the highly organized collagen fibers in tendons and ligaments when they are orientated at a 55° angle to the main magnetic field. We present several examples from a clinical practice setting using 3T imaging illustrating a substantial reduction in magic angle artifact of the peroneal tendon in the prone plantarflexed position compared with the standard neutral (right angle) position. PMID:26429616

  7. Advances in the continuous monitoring of erosion and deposition dynamics: Developments and applications of the new PEEP-3T system

    NASA Astrophysics Data System (ADS)

    Lawler, D. M.

    2008-01-01

    In most episodic erosion and deposition systems, knowledge of the timing of geomorphological change, in relation to fluctuations in the driving forces, is crucial to strong erosion process inference, and model building, validation and development. A challenge for geomorphology, however, is that few studies have focused on geomorphological event structure (timing, magnitude, frequency and duration of individual erosion and deposition events), in relation to applied stresses, because of the absence of key monitoring methodologies. This paper therefore (a) presents full details of a new erosion and deposition measurement system — PEEP-3T — developed from the Photo-Electronic Erosion Pin sensor in five key areas, including the addition of nocturnal monitoring through the integration of the Thermal Consonance Timing (TCT) concept, to produce a continuous sensing system; (b) presents novel high-resolution datasets from the redesigned PEEP-3T system for river bank system of the Rivers Nidd and Wharfe, northern England, UK; and (c) comments on their potential for wider application throughout geomorphology to address these key measurement challenges. Relative to manual methods of erosion and deposition quantification, continuous PEEP-3T methodologies increase the temporal resolution of erosion/deposition event detection by more than three orders of magnitude (better than 1-second resolution if required), and this facility can significantly enhance process inference. Results show that river banks are highly dynamic thermally and respond quickly to radiation inputs. Data on bank retreat timing, fixed with PEEP-3T TCT evidence, confirmed that they were significantly delayed up to 55 h after flood peaks. One event occurred 13 h after emergence from the flow. This suggests that mass failure processes rather than fluid entrainment dominated the system. It is also shown how, by integrating turbidity instrumentation with TCT ideas, linkages between sediment supply and sediment

  8. Catechin and quercetin attenuate adipose inflammation in fructose-fed rats and in 3T3-L1 adipocytes

    PubMed Central

    Vazquez Prieto, Marcela A.; Bettaieb, Ahmed; Rodriguez Lanzi, Cecilia; Soto, Verónica C.; Perdicaro, Diahann J.; Galmarini, Claudio R.; Haj, Fawaz G.; Miatello, Roberto M.; Oteiza, Patricia I.

    2015-01-01

    Scope This study evaluated the capacity of dietary catechin (C), quercetin (Q) and the combination of both (CQ), to attenuate adipose inflammation triggered by high fructose (HFr) consumption in rats and by tumor necrosis factor alpha (TNFα) in 3T3-L1 adipocytes. Methods and results In rats, HFr consumption for 6 wk caused dyslipidemia, insulin resistance, reduced plasma adiponectin, adiposity, and adipose tissue inflammation. Dietary supplementation with 20 mg/kg/d of C, Q and CQ improved all these parameters. In 3T3-L1 adipocytes, C and Q attenuated TNFα-induced elevated protein carbonyls, increased pro-inflammatory cytokine expression (MCP-1, resistin), and decreased adiponectin. The protective effects of C and Q on adipose inflammation are in part associated with their capacity to: i) decrease the activation of the mitogen activated kinases (MAPKs) JNK and p38; and ii) prevent the downregulation of PPARγ. In summary, C and Q, and to a larger extent the combination of both, attenuated adipose pro-inflammatory signaling cascades and regulated the balance of molecules that improve (adiponectin) or impair (TNFα, MCP-1, resistin) insulin sensitivity. Conclusion Together, these findings suggest that dietary Q and C may have potential benefits in mitigating MetS associated adipose inflammation, oxidative stress, and insulin resistance. PMID:25620282

  9. Restoration of murine femoral segmental defect using CTGF-overexpressing MC3T3-E1 cells

    PubMed Central

    Huang, Xiangyu; Li, Yanqiu; Xu, Jiantao; Liu, Kai; Yu, Xin; Cheng, Xin; Xu, Dongdong; Li, Zubing

    2016-01-01

    Connective tissue growth factor (CTGF) is a member of the CCN super family and is reported to widely participate in bone development and regeneration. This study aimed to restore murine femoral segmental defect using CTGF-overexpressing MC3T3-E1 cells. MC3T3-E1 cells were transinfected by lenti-CTGF (LvCTGF) and lenti-negative control (LvNC) virus to obtain stably transinfected cells. Real-time PCR, Western blot, alkaline phosphatase activity assay, and alizarin red staining demonstrated that the overexpression of CTGF enhanced osteogenesis in vitro. Cell migration assay results showed that LvCTGF cells expressed higher migration ability than LvNC cells, while CCK-8 assay revealed no significant difference in cell proliferation. The LvCTGF and LvNC cells were then seeded into a chitosan/β-TCP scaffold and were used to restore a murine femoral segmental defect. Samples were harvested by the end of 2 and 5 weeks respectively. Micro-CT analysis and Masson’s trichrome staining results showed that the LvCTGF-scaffold group expressed better bone healing compared with the LvNC-scaffold and scaffold-only groups. CTGF-overexpressed cells serve as an efficient source of seeding cells for bone regeneration. PMID:27186279

  10. Ex-PRESS glaucoma filter: an MRI compatible metallic orbital foreign body imaged at 1.5 and 3 T

    PubMed Central

    Mabray, M.C.; Uzelac, A.; Talbott, J.F.; Lin, S.C.; Gean, A.D.

    2015-01-01

    AIM To report on the MRI compatibility of the Ex-PRESS glaucoma filtration device, a tiny metallic implant placed into the anterior chamber of the eye that is much smaller than traditional glaucoma shunts, and to educate the radiology community regarding its appearance. MATERIALS AND METHODS Seven patients with Ex-PRESS glaucoma filtration devices were identified that had undergone MRI at San Francisco General Hospital/University of California San Francisco Medical Center by searching and cross-referencing the radiology reporting system and the electronic medical record. MRI images were reviewed for artefact interfering with interpretation. Ophthalmology examinations were reviewed for evidence of complications. RESULTS Eighteen individual MRI examinations were performed during 12 unique MRI events on these 7 patients. 13/18 individual MRI examinations and 7/12 MRI events were performed at 3 T with the others performed at 1.5 T. Mean time from Ex-PRESS implantation to MRI was 17.5 months. Mean time from MRI to first ophthalmology examination was 1.1 months and from MRI to latest ophthalmology examination was 6.6 months. Susceptibility artefact did not interfere with image interpretation and no complications related to MRI were encountered. CONCLUSION The Ex-PRESS glaucoma filtration device appears to be safe for MRI at 1.5 and 3 T and does not produce significant susceptibility artefact to affect diagnostic interpretation adversely. PMID:25735675

  11. CD4+CXCR3+ T cells and plasmacytoid dendritic cells drive accelerated atherosclerosis associated with systemic lupus erythematosus.

    PubMed

    Clement, Marc; Charles, Nicolas; Escoubet, Brigitte; Guedj, Kevin; Chauveheid, Marie-Paule; Caligiuri, Giuseppina; Nicoletti, Antonino; Papo, Thomas; Sacre, Karim

    2015-09-01

    Cardiovascular disease due to accelerated atherosclerosis is the leading cause of death in patients with systemic lupus erythematosus (SLE). Noteworthy, accelerated atherosclerosis in SLE patients appears to be independant of classical Framingham risk factors. This suggests that aggravated atherosclerosis in SLE patients may be a result of increased inflammation and altered immune responses. However, the mechanisms that mediate the acceleration of atherosclerosis in SLE remain elusive. Based on experimental data which includes both humans (SLE patients and control subjects) and rodents (ApoE-/- mice), we herein propose a multi-step model in which the immune dysfunction associated with SLE (i.e. high level of IFN-α production by TLR 9-stimulated pDCs) is associated with, first, an increased frequency of circulating pro inflammatory CD4+CXCR3+ T cells; second, an increased production of CXCR3 ligands by endothelial cells; third, an increased recruitment of pro-inflammatory CD4+CXCR3+ T cells into the arterial wall, and fourth, the development of atherosclerosis. In showing how SLE may promote accelerated atherosclerosis, our model also points to hypotheses for potential interventions, such as pDCs-targeted therapy, that might be studied in the future. PMID:26183767

  12. Berberine inhibits SREBP-1-related clozapine and risperidone induced adipogenesis in 3T3-L1 cells.

    PubMed

    Hu, Yueshan; Kutscher, Eric; Davies, Gareth E

    2010-12-01

    Weight gain is a common and potentially serious complication associated with the treatment of second generation antipsychotics such as clozapine and risperidone. Increased peripheral adipogenesis via the SREBP-1 pathway could be one critical mechanism responsible for antipsychotic drug-induced weight gain. Berberine, a botanical alkaloid, has been shown in our previous studies to inhibit adipogenesis in cell and animal models. MTT was used to determine the cytotoxic effects of clozapine and risperidone in combination with berberine. Differentiation of 3T3-L1 cells was monitored by Oil-Red-O staining and the expression of SREBP-1 and related proteins was determined by real-time RT-PCR and western blotting. The results showed that neither clozapine nor risperidone, alone or in combination with berberine had significant effects on cell viability. Eight days treatment with 15 μM clozapine increased adipogenesis by 37.4% and 50 μM risperidone increased adipogenesis by 26.5% during 3T3-L1 cell differentiation accompanied by increased SREBP-1, PPARγ, C/EBPα, LDLR and Adiponectin gene expression. More importantly, the addition of 8 μM berberine diminished the induction of adipogenesis almost completely accompanied by down-regulated mRNA and protein expression levels of SREBP-1-related proteins. These encouraging results may lead to the use of berberine as an adjuvant to prevent weight gain during second generation antipsychotic medication. PMID:20564506

  13. Synthesis of branched chains with actuation redundancy for eliminating interior singularities of 3T1R parallel mechanisms

    NASA Astrophysics Data System (ADS)

    Li, Shihua; Liu, Yanmin; Cui, Hongliu; Niu, Yunzhan; Zhao, Yanzhi

    2016-03-01

    Although it is common to eliminate the singularity of parallel mechanism by adding the branched chain with actuation redundancy, there is no theory and method for the configuration synthesis of the branched chain with actuation redundancy in parallel mechanism. Branched chains with actuation redundancy are synthesized for eliminating interior singularity of 3-translational and 1- rotational(3T1R) parallel mechanisms. Guided by the discriminance method of hybrid screw group according to Grassmann line geometry, all the possibilities are listed for the occurrence of interior singularities in 3T1R parallel mechanism. Based on the linear relevance of screw system and the principles of eliminating parallel mechanism singularity with actuation redundancy, different types of branched chains with actuation redundancy are synthesized systematically to indicate the layout and the number of the branched chainsinterior with actuation redundancy. A general method is proposed for the configuration synthesis of the branched chains with actuation redundancy of the redundant parallel mechanism, and it builds a solid foundation for the subsequent performance optimization of the redundant actuation parallel mechanism.

  14. Characterization of RNA from Noninfectious Virions Produced by Sarcoma Positive-Leukemia Negative Transformed 3T3 Cells

    PubMed Central

    Phillips, Leo A.; Hollis, Vincent W.; Bassin, Robert H.; Fischinger, Peter J.

    1973-01-01

    RNA from noninfectious virions produced by two established clonal lines of sarcoma positive-leukemia negative (S+L-)-transformed 3T3 cells has been characterized. RNA from virions or nucleoids of S+L--(C243) cells consisted of three to four sizes: ±44 S (6%), 28 S (17%), 18 S (38%), and <18 S (39%). 28S virion RNA contained some virus-specific information demonstrable by RNA·DNA hybridization with a DNA probe derived from the RNA-directed DNA polymerase product of murine sarcoma-leukemia virus, while parallel studies indicated that 28S ribosomal RNA from ribosomal subunits of transformed and nontransformed 3T3 cells did not contain virus-specific information. In contrast to the S+L-(C243) virions, RNA from virions or nucleoids of S+L-(D56) cells consisted of five sizes: 80 S (6%), 68 S (8%), 56 S (5%), 28 S (28%), and <28 S (53%). Thermal dissociation studies suggested that this S+L- genome is comprised of 28S RNA subunits. From these studies we postulate that the 28S viral RNA is most probably the monomeric genome of S+L- virions. PMID:4355380

  15. Anti-obesity and antioxidative effects of purple sweet potato extract in 3T3-L1 adipocytes in vitro.

    PubMed

    Ju, Jae-Hyun; Yoon, Hong-Sup; Park, Hyun-Joon; Kim, Mi-Young; Shin, Hyeun-Kil; Park, Kun-Young; Yang, Jin-Oh; Sohn, Min-Shik; Do, Myoung-Sool

    2011-10-01

    The purpose of the current study was to determine the anti-obesity and anti-inflammatory effects of an extract of purple sweet potatoes (PSPs) on 3T3-L1 adipocytes. For this purpose, differentiated 3T3-L1 adipocytes were treated with a PSP extract at concentrations of 1,000, 2,000, and 3,000 μg/mL for 24 hours. Then, we measured the changes in the sizes of the adipocytes, the secretion of leptin, and the mRNA/protein expression of lipogenic, inflammatory, and lipolytic factors after the treatment with the PSP extract. The PSP extract diminished leptin secretion, indicating that growth of fat droplets was suppressed. The extract also suppressed the expression of mRNAs of lipogenic and inflammatory factors and promoted lipolytic action. The antioxidative activity of the PSP extract was also measured using three different in vitro methods: 1,1-diphenyl-2-picrylhydrazyl free radical scavenging activity, ferric reducing ability potential assay, and chelating activity of transition metal ions. Taken together, our study shows that PSP extract has antilipogenic, anti-inflammatory, and lipolytic effects on adipocytes and has radical scavenging and reducing activity. PMID:21861722

  16. Effects of Scytosiphon lomentaria on osteoblastic proliferation and differentiation of MC3T3-E1 cells

    PubMed Central

    Park, Mi Hwa; Kim, Seoyeon; Cheon, Jihyeon; Lee, Juyeong; Kim, Bo Kyung; Lee, Sang-Hyeon; Kong, Changsuk; Kim, Yuck Yong

    2016-01-01

    BACKGROUND/OBJECTIVES Bone formation and bone resorption continuously occur in bone tissue to prevent the accumulation of old bone, this being called bone remodeling. Osteoblasts especially play a crucial role in bone formation through the differentiation and proliferation. Therefore, in this study, we investigated the effects of Scytosiphon lomentaria extract (SLE) on osteoblastic proliferation and differentiation in MC3T3-E1 cells. MATERIALS/METHODS A cell proliferation assay, alkaline phosphatase (ALP) activity assay, alizarin red staining and protein expression analysis of osteoblastic genes were carried out to assess the osteoblastic proliferation and differentiation. RESULTS The results indicated that treatment of SLE promoted the proliferation of MC3T3-E1 cells and improved ALP activity. And, SLE treatment significantly promoted mineralized nodule formation compared with control. In addition, cells treated with SLE significantly upregulated protein expression of ALP, type 1 collagen, bone morphogenetic protein 2, runt-related transcription factor 2, osterix, and osteoprotegerin. CONCLUSIONS The results demonstrate that SLE promote differentiation inducement and proliferation of osteoblasts and, therefore may help to elucidate the transcriptional mechanism of bone formation and possibly lead to the development of bone-forming drugs. PMID:27087897

  17. Suppression of lipin-1 expression increases monocyte chemoattractant protein-1 expression in 3T3-L1 adipocytes

    SciTech Connect

    Takahashi, Nobuhiko; Hiranaka, Natsumi; Suzuki, Takeshi; Yui, Tomoo; Akanuma, Masayasu; Oka, Kazuya; Kanazawa, Kaoru; Yoshida, Mika; Naito, Sumiyoshi; Fujiya, Mikihiro; Kohgo, Yutaka

    2011-11-11

    Highlights: Black-Right-Pointing-Pointer Lipin-1 affects lipid metabolism, adipocyte differentiation, and transcription. Black-Right-Pointing-Pointer Adipose lipin-1 expression is reduced in obesity. Black-Right-Pointing-Pointer Lipin-1 depletion using siRNA in 3T3-L1 adipocytes increased MCP-1 expression. Black-Right-Pointing-Pointer Lipin-1 is involved in adipose inflammation. -- Abstract: Lipin-1 plays a crucial role in the regulation of lipid metabolism and cell differentiation in adipocytes. Expression of adipose lipin-1 is reduced in obesity, and metabolic syndrome. However, the significance of this reduction remains unclear. This study investigated if and how reduced lipin-1 expression affected metabolism. We assessed mRNA expression levels of various genes related to adipocyte metabolism in lipin-1-depleted 3T3-L1 adipocytes by introducing its specific small interfering RNA. In lipin-1-depleted adipocytes, mRNA and protein expression levels of monocyte chemoattractant protein-1 (MCP-1) were significantly increased, although the other genes tested were not altered. The conditioned media from the cells promoted monocyte chemotaxis. The increase in MCP-1 expression was prevented by treatment with quinazoline or salicylate, inhibitors of nuclear factor-{kappa}B activation. Because MCP-1 is related to adipose inflammation and systemic insulin resistance, these results suggest that a reduction in adipose lipin-1 in obesity may exacerbate adipose inflammation and metabolism.

  18. Macrophage-conditioned medium inhibits differentiation-induced Rb phosphorylation in 3T3-L1 preadipocytes

    SciTech Connect

    Yarmo, Michelle N.; Landry, Anne; Molgat, Andre S.D.; Gagnon, AnneMarie; Sorisky, Alexander

    2009-02-01

    This study examines the mechanisms underlying the anti-adipogenic effect of macrophage-secreted products. 3T3-L1 preadipocytes were induced to differentiate over 8 days in medium conditioned by murine J774 macrophages (MacCM). The inhibitory effect on lipid accumulation and expression of adipogenic markers was diminished when addition of MacCM was delayed to day 2 of differentiation. Clonal expansion, an early event required for 3T3-L1 adipogenesis, was reduced in the presence of MacCM (89%; n = 3; p < 0.001), and BrdU incorporation was impaired by 55% (n = 3; p < 0.01). Activation of ERK1/2 was not affected by MacCM, and neither was the expression of p27{sup kip1}, a cyclin-dependent kinase inhibitor. However, phosphorylation of the retinoblastoma protein (Rb), required for cell cycle progression, was impaired by MacCM (94% inhibition; n = 3; p < 0.01). Differentiation-dependent expression, nuclear localization, and DNA binding ability of C/EBP{beta} were not inhibited by MacCM. Alterations in cell cycle-associated proteins may be important with respect to the anti-adipogenic action of MacCM.

  19. Effect of bisphenol A on SOCS-3 and insulin signaling transduction in 3T3-L1 adipocytes.

    PubMed

    Dai, Yue-E; Chen, Wei; Qi, Humin; Liu, Qian-Qi

    2016-07-01

    The aim of the present study was to investigate whether environmental endocrine disrupting chemical, bisphenol A (BPA), affects secretion of suppressor of cytokine signaling 3 (SOCS-3) and insulin signaling transduction in 3T3-L1 adipocytes. 3T3-L1 adipocytes were treated for 0, 2, 6, 12 and 24 h with BPA at 80 µM in serum‑deprived medium. Reverse transcription-quantitative polymerase chain reaction and western blotting were performed to detect the mRNA expression levels of SOCS‑3 and protein expression levels of SOCS‑3, insulin receptor substrate 1 (IRS‑1), phosphorylated (p)‑IRS‑1, Akt and p‑Akt. The levels of p‑IRS‑1, Akt and p‑Akt in cultures treated for 6 h with BPA were also analyzed by immunofluorescence. The SOCS‑3 mRNA and protein expression levels were decreased in the 6, 12 and 24 h groups. The levels of p‑IRS‑1 and p‑Akt protein were markedly downregulated, while the level of IRS‑1 and Akt protein remained unaltered among these groups, which was consistent with the results observed using immunofluorescence. BPA may inhibit insulin signal transduction and result in the occurrence of insulin resistance via promoting the expression of SOCS-3. PMID:27176707

  20. 5,6-Dehydrokawain from Alpinia zerumbet promotes osteoblastic MC3T3-E1 cell differentiation.

    PubMed

    Kumagai, Momochika; Mishima, Takashi; Watanabe, Akio; Harada, Teppei; Yoshida, Izumi; Fujita, Kazuhiro; Watai, Masatoshi; Tawata, Shinkichi; Nishikawa, Keisuke; Morimoto, Yoshiki

    2016-07-01

    Bone homeostasis is maintained by balancing bone formation and bone resorption, but an imbalance between them is associated with various bone-related diseases such as osteoporosis and rheumatoid arthritis. We found that 5,6-dehydrokawain (DK) and dihydro-5,6-dehydrokawain (DDK), which were isolated as promising compounds from Alpinia zerumbet rhizomes, promote differentiation of osteoblastic MC3T3-E1 cells. DK and DDK increased the alkaline phosphatase activity and matrix mineralization of MC3T3-E1 cells. DK exerts larger effects than DDK. The gene expression of runt-related transcription factor 2 and osterix, which are essential transcription factors in the early period of osteoblast differentiation, was significantly increased by DK treatment. The mRNA level of distal-less homeobox 5 was also enhanced by DK treatment, and DK activated the p38 mitogen-activated protein kinase pathway. Therefore, DK may have clinical potential for preventing osteoporosis, and could be considered as a potential anabolic therapeutic agent. PMID:26940726