WorldWideScience

Sample records for model t-3 gem-t3

  1. A geopotential model from satellite tracking, altimeter, and surface gravity data: GEM-T3

    Science.gov (United States)

    Lerch, F. J.; Nerem, R. S.; Putney, B. H.; Felsentreger, T. L.; Sanchez, B. V.; Marshall, J. A.; Klosko, S. M.; Patel, G. B.; Williamson, R. G.; Chinn, D. S.

    1994-01-01

    An improved model of Earth's gravitational field, Goddard Earth Model T-3 (GEM-T3), has been developed from a combination of satellite tracking, satellite altimeter, and surface gravimetric data. GEM-T3 provides a significant improvement in the modeling of the gravity field at half wavelengths of 400 km and longer. This model, complete to degree and order 50, yields more accurate satellite orbits and an improved geoid representation than previous Goddard Earth Models. GEM-T3 uses altimeter data from GEOS 3 (1975-1976), Seasat (1978) and Geosat (1986-1987). Tracking information used in the solution includes more than 1300 arcs of data encompassing 31 different satellites. The recovery of the long-wavelength components of the solution relies mostly on highly precise satellite laser ranging (SLR) data, but also includes Tracking Network (TRANET) Doppler, optical, and satellite-to-satellite tracking acquired between the ATS 6 and GEOS 3 satellites. The main advances over GEM-T2 (beyond the inclusion of altimeter and surface gravity information which is essential for the resolution of the shorter wavelength geoid) are some improved tracking data analysis approaches and additional SLR data. Although the use of altimeter data has greatly enhanced the modeling of the ocean geoid between 65 deg N and 60 deg S latitudes in GEM-T3, the lack of accurate detailed surface gravimetry leaves poor geoid resolution over many continental regions of great tectonic interest (e.g., Himalayas, Andes). Estimates of polar motion, tracking station coordinates, and long-wavelength ocean tidal terms were also made (accounting for 6330 parameters). GEM-T3 has undergone error calibration using a technique based on subset solutions to produce reliable error estimates. The calibration is based on the condition that the expected mean square deviation of a subset gravity solution from the full set values is predicted by the solutions' error covariances. Data weights are iteratively adjusted until

  2. T3 and cardiac myocyte cell: a theoretical model.

    Science.gov (United States)

    Athanasios, Tsatsaris; Antonios, Baldoukas; Antonios, Loumousiotis; Eustathios, Koukounaris; Maria, Giota; Despina, Perrea

    2013-08-01

    In the last decades, the outstanding role of Thyroid gland in regulating both physiological and pathological operation of cardiovascular system has been acknowledged worldwide. Three main domains of Thyroid function, that is to say, euthyroidism -hyperthyroidism-hypothyroidism, have a direct impact on cardiac response through a variety of mechanisms. Cellular pathways mediate in cardiac contractility, cardiac output, cardiac rhythm, arterial blood pressure and peripheral vessel resistance. Particular biochemical algorithms exist not only between Thyroid hormones' serum concentration and thyroid gland but also between the hormones' serum level and heart muscle genes. These biochemical pathways primarily regulate the appropriate secretion of levothyroxine (T4) and triiodothyronine(T3) via Thyroid- Stimulating-Hormone(TSH) pituitary system, and secondly adjust the cardiac function. In this study, a mathematic model has been developed describing significant aspects of positive or negative feedback mechanisms of THYRO-CARDIAC (THY-CAR) system along with potential applications of novel up-to-date patents in this area of research.

  3. Magnetic properties of the α -T3 model: Magneto-optical conductivity and the Hofstadter butterfly

    Science.gov (United States)

    Illes, E.; Nicol, E. J.

    2016-09-01

    The α -T3 model interpolates between the pseudospin S =1 /2 honeycomb lattice of graphene and the pseudospin S =1 dice lattice via parameter α . We present calculations of the magnetic properties of this hybrid pseudospin model, namely the absorptive magneto-optical conductivity and the Hofstadter butterfly spectra. In the magneto-optics curves, signatures of the hybrid system include a doublet structure present in the peaks, resulting from differing Landau level energies in the K and K' valleys. In the Hofstadter spectra, we detail the evolution of the Hofstadter butterfly as it changes its periodicity by a factor of three as we vary between the two limiting cases of the α -T3 model.

  4. A heat capacity model of T(3/2) dependence for quantum dots.

    Science.gov (United States)

    Roslee, Amirul Edham; Muzakir, Saifful Kamaluddin; Ismail, Jamil; Yusoff, Mashitah M; Jose, Rajan

    2016-12-21

    This article addresses the heat capacity of quantum dots (QDs) using density functional theory (DFT). By analyzing the evolution of phonon density of states and heat capacity as CdSe clusters grow from a molecular cluster into larger quantum confined solids, we have shown that their heat capacity does not fit very well with the Debye T(3) model. We observed that the number of phonon modes, which is discrete, increases as the particles grow, and the dispersion relation shows a quadratic behavior in contrast to the bulk solids whose dispersion relation is linear and equal to the sound velocity. The phonon density of states showed a square root variation with respect to frequency whereas that of the bulk is a quadratic variation of frequency. From the observed variation in the phonon density of states and holding the fact that the atomic vibrations in solids are elastically coupled, we have re-derived the expression for total energy of the QDs and arrive at a T(3/2) model of heat capacity, which fits very well to the observed heat capacity data. These results give promising directions in the understanding of the evolution of the thermophysical properties of solids.

  5. Uniqueness of the Fock quantization of the Gowdy $T^3$ model

    CERN Document Server

    Cortez, J; Velhinho, J M; Cortez, Jeronimo; Marugan, Guillermo A. Mena; Velhinho, Jose M.

    2007-01-01

    After its reduction by a gauge-fixing procedure, the family of linearly polarized Gowdy $T^3$ cosmologies admit a scalar field description whose evolution is governed by a Klein-Gordon type equation in a flat background in 1+1 dimensions with the spatial topology of $S^1$, though in the presence of a time-dependent potential. The model is still subject to a homogeneous constraint, which generates $S^1$-translations. Recently, a Fock quantization of this scalar field was introduced and shown to be unique under the requirements of unitarity of the dynamics and invariance under the gauge group of $S^1$-translations. In this work, we extend and complete this uniqueness result by considering other possible scalar field descriptions, resulting from reasonable field reparameterizations of the induced metric of the reduced model. In the reduced phase space, these alternate descriptions can be obtained by means of a time-dependent scaling of the field, the inverse scaling of its canonical momentum, and the possible ad...

  6. 3D Finite Element Modelling of Drilling Process of Al2024-T3 Alloy with solid tooling and Experimental Validation

    DEFF Research Database (Denmark)

    Davoudinejad, Ali; Tosello, Guido

    2017-01-01

    Drilling is an indispensable process for many manufacturing industries due to its importance for assembling components. This study presents a 3D finite element modelling (3D FEM) approach for drilling process of aluminium 2024-T3. The 3D model of drilling tools for two facet HSSCo and four facet...

  7. 3D Finite Element Modelling of Drilling Process of Al2024-T3 Alloy with solid tooling and Experimental Validation

    DEFF Research Database (Denmark)

    Davoudinejad, Ali; Tosello, Guido

    Drilling is an indispensable process for many manufacturing industries due to the importance of the process for assembling components. This study presents a 3D finite element modeling (3D FEM) approach for drilling process of aluminum 2024-T3. The 3D model of tool for two facet HSSCo and four facet...

  8. Numerical modeling of AA2024-T3 friction stir welding process for residual stress evaluation, including softening effects

    DEFF Research Database (Denmark)

    Sonne, Mads Rostgaard; Carlone, Pierpaolo; Palazzo, Gaetano S.;

    2014-01-01

    In the present paper, a numerical finite element model of the precipitation hardenable AA2024-T3 aluminum alloy, consisting of a heat transfer analysis based on the Thermal Pseudo Mechanical model for heat generation, and a sequentially coupled quasi-static stress analysis is proposed. Metallurgi...

  9. Experimental investigations of material models for Ti-6A1-4V and 2024-T3

    Energy Technology Data Exchange (ETDEWEB)

    Leseur, D

    1999-05-03

    This report describes studies of the deformation and failure behavior of Ti-6Al-4V and 2024-T3 aluminum. Data was obtained at high strain rates and large strains using the split Hopkinson pressure bar technique. This information, plus additional data from the literature, was used to critically evaluate the ability of the Johnson Cook material model to represent the deformation and failure response of Ti-6AMV and 2024-T3 under conditions relevant to simulations of engine containment and the influence of uncontained engine debris on aircraft structures. This model is being used in the DYNA3D finite element code, which is being developed/validated for evaluating aircraft/engine designs relative to the federal airworthiness standards and for improving mitigation/containment technology. The results of the experimental work reported here were used to define a new set of material constants for the strength component of the Johnson Cook model for Ti-6Al-4V and 2024-T3. The capabilities and limitations of the model are reviewed. The model can accurately represent the stress-strain response of the materials. The major concern with the Johnson Cook material model is its ability to accurately represent the stress - strain rate response at strain rates greater than 10{sup 3}-10{sup 4} s{sup {minus}1}. Additional work is also needed to adequately account for failure via shear localization, which was the dominant failure mode at high strain rates in both materials. Failure modeling in both Ti-6Al-N and 2024-T3 will be considered further in future reports.

  10. The effect of hardening laws and thermal softening on modeling residual stresses in FSW of aluminum alloy 2024-T3

    DEFF Research Database (Denmark)

    Sonne, Mads Rostgaard; Tutum, Cem Celal; Hattel, Jesper Henri;

    2013-01-01

    or kinematic hardening together with the metallurgical softening model were applied in order to give a first impression of the tendencies in residual stresses in friction stir welds when choosing different hardening and softening behaviors. Secondly, real friction stir butt welding of aluminum alloy 2024-T3...... were simulated and compared with experimentally obtained results for both temperatures and residual stresses (using the slitting method). The comparisons showed good agreement regarding temperatures whereas the residual stress comparisons indicated different sensitivities for the cold and hot welding...

  11. Review and Study of Physics Driven Pitting Corrosion Modeling in 2024-T3 Aluminum Alloys (Postprint)

    Science.gov (United States)

    2015-05-01

    measuring the maximum pit size and its effect on the fatigue life. Microstructure and corrosion currents were neglected. In this paper we present the...Figure 2 Appearance of the pits on the surface of 7075 -T6 (Wang et al., 2003) Proc. of SPIE Vol. 9437 94372E-2 2 Distribution Statement A. Approved... density (Harlow and Wei, 1994). It can be seen that the definition of pitting current Ip is the key element in the pitting corrosion modeling, which

  12. Loganin inhibits the inflammatory response in mouse 3T3L1 adipocytes and mouse model.

    Science.gov (United States)

    Li, Yang; Li, Zheng; Shi, Lei; Zhao, Chenxu; Shen, Bingyu; Tian, Ye; Feng, Haihua

    2016-07-01

    Atherosclerosis is a chronic inflammatory disease of the vascular walls. ApoCIII is an independent factor which promotes atherosclerotic processes. This study aimed to investigate whether Loganin administration inhibits the inflammatory response in vitro and in vivo. In the apoCIII-induced mouse adipocytes, the levels of cytokines, including TNF-α, MCP-1 and IL-6 were determined by enzyme-linked immunosorbent assay and their gene expressions were measured through RT-PCR. The phosphorylation of nuclear factor-κB (NF-κB) proteins was analyzed by Western blotting. Our results showed that Loganin markedly decreased TNF-α, MCP-1 and IL-6 concentrations as well as their gene expressions. Western blotting analysis indicated that Loganin suppressed the activation of NF-κB signaling. In the Tyloxapol-treated mouse model, Loganin reduced the contents of TC and TG in mouse serum. The results of Oil Red-O Staining showed that Loganin reduced the production of lipid droplets. So it is suggested that Loganin might be a potential therapeutic agent for preventing the inflammation stress in vitro and in vivo.

  13. Modelling the residual stresses and microstructural evolution in Friction Stir Welding of AA2024-T3 including the Wagner-Kampmann precipitation model

    DEFF Research Database (Denmark)

    Sonne, Mads Rostgaard; Hattel, Jesper Henri

    In this work, a numerical finite element model for friction stir welding of 2024-T3 aluminum alloy, consisting of a heat transfer analysis and a sequentially coupled quasi-static stress analysis is proposed. Metallurgical softening of the material is properly considered and included...

  14. Performance of an Optimized Eta Model Code on the Cray T3E and a Network of PCs

    Science.gov (United States)

    Kouatchou, Jules; Rancic, Miodrag; Geiger, Jim

    2000-01-01

    In the year 2001, NASA will launch the satellite TRIANA that will be the first Earth observing mission to provide a continuous, full disk view of the sunlit Earth. As a part of the HPCC Program at NASA GSFC, we have started a project whose objectives are to develop and implement a 3D cloud data assimilation system, by combining TRIANA measurements with model simulation, and to produce accurate statistics of global cloud coverage as an important element of the Earth's climate. For simulation of the atmosphere within this project we are using the NCEP/NOAA operational Eta model. In order to compare TRIANA and the Eta model data on approximately the same grid without significant downscaling, the Eta model will be integrated at a resolution of about 15 km. The integration domain (from -70 to +70 deg in latitude and 150 deg in longitude) will cover most of the sunlit Earth disc and will continuously rotate around the globe following TRIANA. The cloud data assimilation is supposed to run and produce 3D clouds on a near real-time basis. Such a numerical setup and integration design is very ambitious and computationally demanding. Thus, though the Eta model code has been very carefully developed and its computational efficiency has been systematically polished during the years of operational implementation at NCEP, the current MPI version may still have problems with memory and efficiency for the TRIANA simulations. Within this work, we optimize a parallel version of the Eta model code on a Cray T3E and a network of PCs (theHIVE) in order to improve its overall efficiency. Our optimization procedure consists of introducing dynamically allocated arrays to reduce the size of static memory, and optimizing on a single processor by splitting loops to limit the number of streams. All the presented results are derived using an integration domain centered at the equator, with a size of 60 x 60 deg, and with horizontal resolutions of 1/2 and 1/3 deg, respectively. In accompanying

  15. Murine 3T3-L1 adipocyte cell differentiation model: validated reference genes for qPCR gene expression analysis.

    Directory of Open Access Journals (Sweden)

    Tatjana Arsenijevic

    Full Text Available BACKGROUND: Analysis of gene expression at the mRNA level, using real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR, mandatorily requires reference genes (RGs as internal controls. However, increasing evidences have shown that RG expression may vary considerably under experimental conditions. We sought for an appropriate panel of RGs to be used in the 3T3-L1 cell line model during their terminal differentiation into adipocytes. To this end, the expression levels of a panel of seven widely used RG mRNAs were measured by qRT-PCR. The 7 RGs evaluated were ß-actin (ACTB, glyceraldehyde-3-phosphate dehydrogenase (GAPDH, hypoxanthine phosphoribosyl-transferase I (HPRT, ATP synthase H+ transporting mitochondrial F1 complex beta subunit (ATP-5b, tyrosine 3-monooxygenase/tryptophan 5- monooxygenase activation protein, zeta polypeptide (Ywhaz, Non-POU-domain containing octamer binding protein (NoNo, and large ribosomal protein L13a (RPL. METHODOLOGY/PRINCIPAL FINDINGS: Using three Excel applications, GeNorm, NormFinder and BestKeeper, we observed that the number and the stability of potential RGs vary significantly during differentiation of 3T3-L1 cells into adipocytes. mRNA expression analyses using qRT-PCR revealed that during the entire differentiation program, only NoNo expression is relatively stable. Moreover, the RG sets that were acceptably stable were different depending on the phase of the overall differentiation process (i.e. mitotic clonal expansion versus the terminal differentiation phase. RPL, ACTB, and Ywhaz, are suitable for terminal differentiation, whereas ATP-5b and HPRT, are suitable during mitotic clonal expansion. CONCLUSION: Our results demonstrate that special attention must be given to the choice of suitable RGs during the various well defined phases of adipogenesis to ensure accurate data analysis and that the use of several RGs is absolutely required. Consequently, our data show for the first time

  16. A thyroid hormone challenge in hypothyroid rats identifies T3 regulated genes in the hypothalamus and in models with altered energy balance and glucose homeostasis.

    Science.gov (United States)

    Herwig, Annika; Campbell, Gill; Mayer, Claus-Dieter; Boelen, Anita; Anderson, Richard A; Ross, Alexander W; Mercer, Julian G; Barrett, Perry

    2014-11-01

    expressed in animal models of long- and short-term body weight regulation. This study identified genes regulated by T3 in the hypothalamus, a key area of the brain involved in homeostasis and neuroendocrine functions. These include genes hitherto not known to be regulated by thyroid status.

  17. An Atmospheric General Circulation Model with Chemistry for the CRAY T3E: Design, Performance Optimization and Coupling to an Ocean Model

    Science.gov (United States)

    Farrara, John D.; Drummond, Leroy A.; Mechoso, Carlos R.; Spahr, Joseph A.

    1998-01-01

    The design, implementation and performance optimization on the CRAY T3E of an atmospheric general circulation model (AGCM) which includes the transport of, and chemical reactions among, an arbitrary number of constituents is reviewed. The parallel implementation is based on a two-dimensional (longitude and latitude) data domain decomposition. Initial optimization efforts centered on minimizing the impact of substantial static and weakly-dynamic load imbalances among processors through load redistribution schemes. Recent optimization efforts have centered on single-node optimization. Strategies employed include loop unrolling, both manually and through the compiler, the use of an optimized assembler-code library for special function calls, and restructuring of parts of the code to improve data locality. Data exchanges and synchronizations involved in coupling different data-distributed models can account for a significant fraction of the running time. Therefore, the required scattering and gathering of data must be optimized. In systems such as the T3E, there is much more aggregate bandwidth in the total system than in any particular processor. This suggests a distributed design. The design and implementation of a such distributed 'Data Broker' as a means to efficiently couple the components of our climate system model is described.

  18. An Atmospheric General Circulation Model with Chemistry for the CRAY T3E: Design, Performance Optimization and Coupling to an Ocean Model

    Science.gov (United States)

    Farrara, John D.; Drummond, Leroy A.; Mechoso, Carlos R.; Spahr, Joseph A.

    1998-01-01

    The design, implementation and performance optimization on the CRAY T3E of an atmospheric general circulation model (AGCM) which includes the transport of, and chemical reactions among, an arbitrary number of constituents is reviewed. The parallel implementation is based on a two-dimensional (longitude and latitude) data domain decomposition. Initial optimization efforts centered on minimizing the impact of substantial static and weakly-dynamic load imbalances among processors through load redistribution schemes. Recent optimization efforts have centered on single-node optimization. Strategies employed include loop unrolling, both manually and through the compiler, the use of an optimized assembler-code library for special function calls, and restructuring of parts of the code to improve data locality. Data exchanges and synchronizations involved in coupling different data-distributed models can account for a significant fraction of the running time. Therefore, the required scattering and gathering of data must be optimized. In systems such as the T3E, there is much more aggregate bandwidth in the total system than in any particular processor. This suggests a distributed design. The design and implementation of a such distributed 'Data Broker' as a means to efficiently couple the components of our climate system model is described.

  19. Chlamydia induces anchorage independence in 3T3 cells and detrimental cytological defects in an infection model.

    Directory of Open Access Journals (Sweden)

    Andrea E Knowlton

    Full Text Available Chlamydia are gram negative, obligate intracellular bacterial organisms with different species causing a multitude of infections in both humans and animals. Chlamydia trachomatis is the causative agent of the sexually transmitted infection (STI Chlamydia, the most commonly acquired bacterial STI in the United States. Chlamydial infections have also been epidemiologically linked to cervical cancer in women co-infected with the human papillomavirus (HPV. We have previously shown chlamydial infection results in centrosome amplification and multipolar spindle formation leading to chromosomal instability. Many studies indicate that centrosome abnormalities, spindle defects, and chromosome segregation errors can lead to cell transformation. We hypothesize that the presence of these defects within infected dividing cells identifies a possible mechanism for Chlamydia as a cofactor in cervical cancer formation. Here we demonstrate that infection with Chlamydia trachomatis is able to transform 3T3 cells in soft agar resulting in anchorage independence and increased colony formation. Additionally, we show for the first time Chlamydia infects actively replicating cells in vivo. Infection of mice with Chlamydia results in significantly increased cell proliferation within the cervix, and in evidence of cervical dysplasia. Confocal examination of these infected tissues also revealed elements of chlamydial induced chromosome instability. These results contribute to a growing body of data implicating a role for Chlamydia in cervical cancer development and suggest a possible molecular mechanism for this effect.

  20. Accepting the T3D

    Energy Technology Data Exchange (ETDEWEB)

    Rich, D.O.; Pope, S.C.; DeLapp, J.G.

    1994-10-01

    In April, a 128 PE Cray T3D was installed at Los Alamos National Laboratory`s Advanced Computing Laboratory as part of the DOE`s High-Performance Parallel Processor Program (H4P). In conjunction with CRI, the authors implemented a 30 day acceptance test. The test was constructed in part to help them understand the strengths and weaknesses of the T3D. In this paper, they briefly describe the H4P and its goals. They discuss the design and implementation of the T3D acceptance test and detail issues that arose during the test. They conclude with a set of system requirements that must be addressed as the T3D system evolves.

  1. In vitro and in vivo study of microporous ceramics using MC3T3 cells, CAM assay and a pig animal model.

    Science.gov (United States)

    Tomco, Marek; Petrovova, Eva; Giretova, Maria; Almasiova, Viera; Holovska, Katarina; Cigankova, Viera; Jenca, Andrej; Jencova, Janka; Jenca, Andrej; Boldizar, Martin; Balazs, Kosa; Medvecky, Lubomir

    2017-09-01

    Bone tissue engineering combines biomaterials with biologically active factors and cells to hold promise for reconstructing craniofacial defects. In this study the biological activity of biphasic hydroxyapatite ceramics (HA; a bone substitute that is a mixture of hydroxyapatite and β-tricalcium phosphate in fixed ratios) was characterized (1) in vitro by assessing the growth of MC3T3 mouse osteoblast lineage cells, (2) in ovo by using the chick chorioallantoic membrane (CAM) assay and (3) in an in vivo pig animal model. Biocompatibility, bioactivity, bone formation and biomaterial degradation were detected microscopically and by radiology and histology. HA ceramics alone demonstrated great biocompatibility on the CAM as well as bioactivity by increased proliferation and alkaline phosphatase secretion of mouse osteoblasts. The in vivo implantation of HA ceramics with bone marrow mesenchymal stem cells (MMSCs) showed de novo intramembranous bone healing of critical-size bone defects in the right lateral side of pig mandibular bodies after 3 and 9 weeks post-implantation. Compared with the HA ceramics without MMSCs, the progress of bone formation was slower with less-developed features. This article highlights the clinical use of microporous biphasic HA ceramics despite the unusually shaped elongated micropores with a high length/width aspect ratio (up to 20) and absence of preferable macropores (>100 µm) in bone regenerative medicine.

  2. T3 may be a better agent than T4 in the critically ill hypothyroid patient: evaluation of transport across the blood-brain barrier in a primate model

    Energy Technology Data Exchange (ETDEWEB)

    Chernow, B.; Burman, K.D.; Johnson, D.L.; McGuire, R.A.; O' Brian, J.T.; Wartofsky, L.; Georges, L.P.

    1983-02-01

    Thyroid hormone transport across the blood brain barrier in hypothyroid patients is clinically important yet poorly understood. To study this question, 200 micrograms of thyroxine (T4), 100 micrograms of 3,5,3'-triiodothyronine (T3) and 100 micrograms of 3,3',5'-triiodothyronine (reverse T3) were administered separately to 3 baboons, first iv and at a later date intrathecally (IT). Six animals were used. Three received the iv injections and three received the IT injections. In each of the 18 experiments, cerebrospinal fluid (CSF) and serum specimens were collected serially for 6 h after injection. Mean maximal elevations from baseline in CSF iodothyronine levels were 100 +/- 10 ng/dl after iv T4, 3921 +/- 293 ng/dl after iv T3 and 31 +/- 17 ng/dl after iv reverse T3. When given IT in the same dosages, the mean maximal increases in serum iodothyronine concentrations were: 1670 +/- 600 ng/dl for T4, 806 +/- 405 ng/dl for T3, and 210 +/- 43 ng/dl for reverse T3. In every animal studied, rapid bidirectional transfer of T3 from serum to CSF and CSF to serum occurred, whereas iv T4 resulted in delayed minimal increments in CSF T4 concentration. Isotopic experiments were also performed and the results analyzed using a kinetic model. When /sup 125/I-T3 was given iv, the equilibrium point in CSF was observed within 90 min with 1.7% of the administered dose/L able to be counted in CSF at any moment in time. When labeled T4 was given iv, only 0.6% of the administered dose/L was counted in CSF and the equilibrium point was not reached until 360 min. These data suggest: T4, T3, and reverse T3 are all capable of bidirectional transfer across the blood brain barrier, T3 may be a better agent than T4 in treating patients with myxedema coma because T3 crosses more rapidly and more completely from serum to CSF.

  3. A Thyroid Hormone Challenge in Hypothyroid Rats Identifies T3 Regulated Genes in the Hypothalamus and in Models with Altered Energy Balance and Glucose Homeostasis

    Science.gov (United States)

    Herwig, Annika; Campbell, Gill; Mayer, Claus-Dieter; Boelen, Anita; Anderson, Richard A.; Ross, Alexander W.; Mercer, Julian G.

    2014-01-01

    Background: The thyroid hormone triiodothyronine (T3) is known to affect energy balance. Recent evidence points to an action of T3 in the hypothalamus, a key area of the brain involved in energy homeostasis, but the components and mechanisms are far from understood. The aim of this study was to identify components in the hypothalamus that may be involved in the action of T3 on energy balance regulatory mechanisms. Methods: Sprague Dawley rats were made hypothyroid by giving 0.025% methimazole (MMI) in their drinking water for 22 days. On day 21, half the MMI-treated rats received a saline injection, whereas the others were injected with T3. Food intake and body weight measurements were taken daily. Body composition was determined by magnetic resonance imaging, gene expression was analyzed by in situ hybridization, and T3-induced gene expression was determined by microarray analysis of MMI-treated compared to MMI-T3-injected hypothalamic RNA. Results: Post mortem serum thyroid hormone levels showed that MMI treatment decreased circulating thyroid hormones and increased thyrotropin (TSH). MMI treatment decreased food intake and body weight. Body composition analysis revealed reduced lean and fat mass in thyroidectomized rats from day 14 of the experiment. MMI treatment caused a decrease in circulating triglyceride concentrations, an increase in nonesterified fatty acids, and decreased insulin levels. A glucose tolerance test showed impaired glucose clearance in the thyroidectomized animals. In the brain, in situ hybridization revealed marked changes in gene expression, including genes such as Mct8, a thyroid hormone transporter, and Agrp, a key component in energy balance regulation. Microarray analysis revealed 110 genes to be up- or downregulated with T3 treatment (±1.3-fold change, phypothalamus, a key area of the brain involved in homeostasis and neuroendocrine functions. These include genes hitherto not known to be regulated by thyroid status. PMID:25087834

  4. Pre-osteoblastic MC3T3-E1 cells promote breast cancer growth in bone in a murine xenograft model

    Institute of Scientific and Technical Information of China (English)

    Thomas M. Bodenstine; Benjamin H. Beck; Xuemei Cao; Leah M. Cook; Aimen Ismai; J. Kent Powers; Andrea M. Mastro; Danny R. Welch

    2011-01-01

    The bones are the most common sites of breast cancer metastasis. Upon arrival within the bone microenvironment, breast cancer cells coordinate the activities of stromal cells, resulting in an increase in osteoclast activity and bone matrix degradation. In late stages of bone metastasis, breast cancer cells induce apoptosis in osteoblasts, which further exacerbates bone loss. However, in early stages, breast cancer cells induce osteoblasts to secrete inflammatory cytokines purported to drive tumor progression. To more thoroughly evaluate the role of osteoblasts in early stages of breast cancer metastasis to the bones, we used green fluorescent protein-labeled human breast cancer cell lines MDA-MB-231 and MDA-MB-435, which both induce osteolysis after intra-femoral injection in athymic mice, and the murine pre-osteoblastic cell line MC3T3-E1 to modulate osteoblast populations at the sites of breast cancer metastasis. Breast cancer cells were injected directly into the femur with or without equal numbers of MC3T3-E1 cells. Tumors grew significantly larger when co-injected with breast cancer cells and MC3T3-E1 cells than injected with breast cancer cells alone. Osteolysis was induced in both groups, indicating that MC3T3-E1 cells did not block the ability of breast cancer cells to cause bone destruction. MC3T3-E1 cells promoted tumor growth out of the bone into the extraosseous stroma. These data suggest that breast cancer cells and osteoblasts communicate during early stages of bone metastasis and promote tumor growth.

  5. $T^3$-interferometer for atoms

    CERN Document Server

    Zimmermann, M; Roura, A; Schleich, W P; DeSavage, S A; Davis, J P; Srinivasan, A; Narducci, F A; Werner, S A; Rasel, E M

    2016-01-01

    The quantum mechanical propagator of a massive particle in a linear gravitational potential derived already in 1927 by Earle H. Kennard \\cite{Kennard,Kennard2} contains a phase that scales with the third power of the time $T$ during which the particle experiences the corresponding force. Since in conventional atom interferometers the internal atomic states are all exposed to the same acceleration $a$, this $T^3$-phase cancels out and the interferometer phase scales as $T^2$. In contrast, by applying an external magnetic field we prepare two different accelerations $a_1$ and $a_2$ for two internal states of the atom, which translate themselves into two different cubic phases and the resulting interferometer phase scales as $T^3$. We present the theoretical background for, and summarize our progress towards experimentally realizing such a novel atom interferometer.

  6. T 3-Interferometer for atoms

    Science.gov (United States)

    Zimmermann, M.; Efremov, M. A.; Roura, A.; Schleich, W. P.; DeSavage, S. A.; Davis, J. P.; Srinivasan, A.; Narducci, F. A.; Werner, S. A.; Rasel, E. M.

    2017-04-01

    The quantum mechanical propagator of a massive particle in a linear gravitational potential derived already in 1927 by Kennard [2, 3] contains a phase that scales with the third power of the time T during which the particle experiences the corresponding force. Since in conventional atom interferometers the internal atomic states are all exposed to the same acceleration a, this T^3-phase cancels out and the interferometer phase scales as T^2. In contrast, by applying an external magnetic field we prepare two different accelerations a_1 and a_2 for two internal states of the atom, which translate themselves into two different cubic phases and the resulting interferometer phase scales as T^3. We present the theoretical background for, and summarize our progress towards experimentally realizing such a novel atom interferometer.

  7. Toxicological evaluation of the topoisomerase inhibitor, etoposide, in the model animal Caenorhabditis elegans and 3T3-L1 normal murine cells.

    Science.gov (United States)

    Lee, So Young; Kim, Joo Yeon; Jung, Yu-Jin; Kang, Kyungsu

    2017-06-01

    Etoposide, a topoisomerase II inhibitor, has been widely used as a clinical anticancer drug to treat diverse cancer patients. Since not only rapidly dividing cancer cells but also the cells of normal human tissues and every living organism in environmental ecosystems have topoisomerases, it is crucial to study the toxicity of etoposide in other organisms in addition to cancer cells. In this study, we evaluated the toxicity of etoposide in both a soil nematode, Caenorhabditis elegans, and 3T3-L1 normal murine cells. Etoposide significantly retarded the growth, egg laying, and hatching in C. elegans. Etoposide also affected the reproductive gonad tissue, decreased the number of germ cells and induced abnormally enlarged nuclei in C. elegans. In addition, etoposide inhibited 3T3-L1 cell proliferation, with IC50 values of 37.8 ± 7.3 and 9.8 ± 1.8 μM after 24 and 48 hours of treatment, respectively, via the induction of cell cycle arrest at the G2/M phase and apoptotic cell death. Etoposide also induced nuclear enlargement in 3T3-L1 normal murine cells. The reproductive toxicity and abnormal nuclear morphological changes seemed to correlate with the adverse effects of etoposide. We suggest that these experimental platforms, i.e., the toxicological evaluation of both nematodes and 3T3-L1 cells, may be useful to study the mechanisms underlying the side effects of chemicals, including topoisomerase inhibitors. © 2017 Wiley Periodicals, Inc.

  8. 钩藤散对NIH-3T3细胞衰老模型增殖与凋亡的影响%Effects of Gouteng San on the Proliferation and Apoptosis of NIH-3T3 Cell Aging Model

    Institute of Scientific and Technical Information of China (English)

    黄厚才; 钟荣玲; 曹鹏; 宋捷; 杨德功; 夏智

    2012-01-01

    目的:观察钩藤散对NIH-3T3细胞衰老模型增殖与凋亡的影响.方法:NIH-3T3细胞接种在含10%胎牛血清的DMEM培养至20代,然后分为年轻、空白、模型3个组,传至30代时,用H2O2处理,β-半乳糖苷酶染色试剂盒染色,观察衰老细胞形态、衰老指征,确认衰老模型是否成功建立;用钩藤散(17,8.5,4.25,2.15 g·kg-1)ig SD大鼠,每天2次,连续给药3d,于末次ig1 h后麻醉、腹主动脉采血,制备含药血清;用钩藤散含药血清处理衰老模型,观察衰老细胞形态、衰老指征、增殖与凋亡等指标.结果:模型组细胞较对照组形态有显著改变,细胞衰老比例明显增加(P<0.05),因此H2O2氧化应激方法可成功建立细胞衰老模型;钩藤散含药血清能明显减少细胞形态的改变、改善衰老指征、促进细胞增殖(P<0.01)、降低细胞凋亡(P<0.01),尤其10倍剂量组最为明显.结论:钩藤散能有效促进细胞增殖,降低细胞凋亡,从而防止衰老.%Objective; To observe the regulatory effect of Gouteng San on the proliferation and apoptosis of NIH-3T3 cell lines. Method; NIH-3T3 cells were cultivated in DMEM with 10% fetal calf serum and three groups such as youth, blank and model group were used. Cells at generation 30 were treated with hydrogen peroxide ( H2 O2) .and then β-galactosidase dyeing kit was applied to observe cell morphology and aging indications to assure whether the aging mode was successfully established. Oral administration of Gouteng San was started for three constitutive days, twice daily. One hour after last administration, rats were anesthetized, and we drew blood from abdominal aorta for the preparation of Gouteng San medicated serum. NIH-3T3 cells were treated with Gouteng San medicated serum to observe cell morphology, aging indications, proliferation and apoptosis. Result; The cell morphology of model was changed and the proportion of cell senescence was increased significantly in model group

  9. The two triiodothyronines (T3 and rT3). Thyroid biosynthesis of T3 and rT3 and peripheral metabolism of thyroxine (author's transl).

    Science.gov (United States)

    Roche, J; Michel, R

    1977-01-01

    Thirty per cent of the iodine in thyroglobulin is present as iodothyronines. L-thyroxine (T4) represents 90-95% of hormonal iodine, 3,5,3'-triodo-L-thyronine (T3) contains at the most two per cent of it, 3,3'5'-triodo-L-thyronine (rT3) even less, as well as traces of 3,3'-diodo-L-thyronine. The plasma concentration of T4 is about 8 microgram per 100 ml, in the case of T3 it is 120 ng and 25 ng for rT3. The cell nucleus preferentially binds T3 and rT3 and there are also some specific mitochondrial proteins which possess a high affinity for T3. L-thyroxine is dehalogenated peripherically to T3, to take care of most of the requirements in T3. The enrichment of the plasma in rT3 has been shown to occur under various experimental and pathological conditoins. The blood level of T3 varies in inverse ratio to the level of rT3 and it shows that the peripheral formation of one is compensated for by the other. The excess of the prehormone T4 is metabolised as 3,5,3',5'-tetraiodothyroacetic acid (TetrAc); its level in the blood varies in the same way as the level of T3, in particular it decreases during starvation.

  10. Effects of berberine on PI-3K p85 protein expression in insulin-resistant cell model in 3T3-L1 adipocytes%小檗碱对3T3-L1胰岛素抵抗细胞模型PI-3K p85蛋白表达的影响

    Institute of Scientific and Technical Information of China (English)

    易屏; 陆付耳; 陈广; 徐丽君; 董慧; 王开富

    2008-01-01

    目的:研究小檗碱对3T3-L1胰岛素抵抗细胞模型PI-3K p85蛋白表达的影响,探讨小檗碱改善胰岛素抵抗的分子机制.方法:分别以0.5 mmol/L软脂酸与25 mmol/L葡萄糖加0.6 nmmol/L胰岛素诱导3T3-L1脂肪细胞产生胰岛素抵抗,予以小檗碱进行干预,同时以阿司匹林作为阳性对照,以2-脱氧-[3H]-D-葡萄糖摄入法观察葡萄糖的转运率,用Western blot检测PI-3K p85蛋白的表达.结果:0.5 mmol/L软脂酸作用24 h或25 mmol/L葡萄糖加0.6 nmmol/L胰岛素作用18 h分别使3T3-L1脂肪细胞胰岛素刺激的葡萄糖转运抑制67%和60%,Westem blot显示PI-3K p85蛋白表达减少,与正常对照组比较有统计学意义(P<0.01);同时加入小檗碱则可逆转上述效应使P1-3K p85蛋白表达增加,与模型组比较有明显差异(P<0.01),并且PI-3K p85蛋白的表达与小檗碱的剂量和作用时间呈依赖关系.结论:小檗碱可以明显改善游离脂肪酸和高糖诱导的胰岛素抵抗,其分子机制可能与小檗碱提高PI-3K p85蛋白的表达有关.

  11. Mitochondrial T3 receptor and targets.

    Science.gov (United States)

    Wrutniak-Cabello, Chantal; Casas, François; Cabello, Gérard

    2017-02-03

    The demonstration that TRα1 mRNA encodes a nuclear thyroid hormone receptor and two proteins imported into mitochondria with molecular masses of 43 and 28 kDa has brought new clues to better understand the pleiotropic influence of iodinated hormones. If p28 activity remains unknown, p43 binds to T3 responsive elements occurring in the organelle genome, and, in the T3 presence, stimulates mitochondrial transcription and the subsequent synthesis of mitochondrial encoded proteins. This influence increases mitochondrial activity and through changes in the mitochondrial/nuclear cross talk affects important nuclear target genes regulating cell proliferation and differentiation, oncogenesis, or apoptosis. In addition, this pathway influences muscle metabolic and contractile phenotype, as well as glycaemia regulation. Interestingly, according to the process considered, p43 exerts opposite or cooperative effects with the well-known T3 pathway, thus allowing a fine tuning of the physiological influence of this hormone. Copyright © 2017 Elsevier B.V. All rights reserved.

  12. Numerical Tokamak Turbulence Calculations on the CRAY T3E

    Energy Technology Data Exchange (ETDEWEB)

    Lynch, V.E., Leboeuf, J.N., Carreras, B.A. [Oak Ridge National Lab., TN (United States)], Alvarez, J.D., Garcia, L. [Universidad `Carlos III` de Madrid (Spain)

    1997-12-31

    Full cross section calculations of ion-temperature-gradient-driven turbulence with Landau closure are being carried out as part of the Numerical Tokamak Turbulence Project, one of the U.S. Department of Energy`s Phase II Grand Challenges. To include the full cross section of a magnetic fusion device like the tokamak requires more memory and CPU time than is available on the National Energy Research Scientific Computing Center`s (NERSC`s) shared-memory vector machines such as the CRAY C90 and J90. Calculations of cylindrical multi-helicity ion-temperature-gradient-driven turbulence were completed on NERSC`s 160-processor distributed-memory CRAY T3E parallel computer with 256 Mbytes of memory per processor. This augurs well for yet more memory and CPU intensive calculations on the next-generation T3E at NERSC. This paper presents results on benchmarks with the current T3E at NERSC. Physics results pertaining to plasma confinement at the core of tokamaks subject to ion-temperature-gradient-driven-turbulence are also highlighted. Results at this resolution covering this extent of physical time were previously unattainable. Work is in progress to increase the resolution, improve the performance of the parallel code, and include toroidal geometry in these calculations in anticipation of the imminent arrival of a fully configured,512-processor, T3E-900 model.

  13. A T3 and T7 recombinant phage acquires efficient adsorption and a broader host range.

    Directory of Open Access Journals (Sweden)

    Tiao-Yin Lin

    Full Text Available It is usually thought that bacteriophage T7 is female specific, while phage T3 can propagate on male and female Escherichia coli. We found that the growth patterns of phages T7M and T3 do not match the above characteristics, instead showing strain dependent male exclusion. Furthermore, a T3/7 hybrid phage exhibits a broader host range relative to that of T3, T7, as well as T7M, and is able to overcome the male exclusion. The T7M sequence closely resembles that of T3. T3/7 is essentially T3 based, but a DNA fragment containing part of the tail fiber gene 17 is replaced by the T7 sequence. T3 displays inferior adsorption to strains tested herein compared to T7. The T3 and T7 recombinant phage carries altered tail fibers and acquires better adsorption efficiency than T3. How phages T3 and T7 recombine was previously unclear. This study is the first to show that recombination can occur accurately within only 8 base-pair homology, where four-way junction structures are identified. Genomic recombination models based on endonuclease I cleavages at equivalent and nonequivalent sites followed by strand annealing are proposed. Retention of pseudo-palindromes can increase recombination frequency for reviving under stress.

  14. Motion of compressible magnetic fluids in T^3

    Directory of Open Access Journals (Sweden)

    Weiping Yan

    2013-10-01

    Full Text Available This article shows the existence of weak time-periodic motion of a three-dimensional system of compressible magnetic fluid driven by time-dependent external forces in a torus T^3. The model consists of the mass conservation equation, the linear momentum equation, the angular momentum equation, the Bloch-Torrey type equation and the magnetostatic equation. This analysis is based on the Faedo-Galerkin method and weak compactness techniques.

  15. Increased Oxidative Stress in Cultured 3T3-L1 Cells was Attenuated by Berberine Treatment.

    Science.gov (United States)

    Dong, Shi-Fen; Yasui, Naomi; Negishb, Hiroko; Kishimoto, Aya; Sun, Jian-Ning; Ikeda, Katsumi

    2015-06-01

    The 3T3-L1 cell line is one of the most well-characterized and reliable models for studying adipocytes. Increased oxidative stress in accumulated fat was found in 3T3-L1 cells. Berberine, an isoquinoline alkaloid, could suppress fat deposition in 3T3-L1 cells; however, whether berberine suppresses increased oxidative stress is not well known. In this study, we observed the effect of berberine on increased oxidative stress in 3T3-L1 cells. 3T3-L1 cells were cultured and treated with berberine (5-20 μM) from day 3 to day 8. We confirmed that berberine markedly inhibited fat accumulation and lipid droplets in 3T3-L1 adipocytes and decreased triglyceride content. Berberine inhibited increased oxidative stress in 3T3-L1 cells by suppressing reactive oxygen species (ROS) production, and increased glutathione peroxidase (GPx) gene expression and GPx activity. Berberine also markedly reduced adipokines secreted by adipocytes, including leptin and resistin.

  16. A characterization of the SPARC T3-4 system

    NARCIS (Netherlands)

    van Tol, M.W.

    2011-01-01

    This technical report covers a set of experiments on the 64-core SPARC T3-4 system, comparing it to two similar AMD and Intel systems. Key characteristics as maximum integer and floating point arithmetic throughput are measured as well as memory throughput, showing the scalability of the SPARC T3-4

  17. Modulation of nuclear T3 binding by T3 in a human hepatocyte cell-line (Chang-liver) - T3 stimulation of cell growth but not of malic enzyme, glucose-6-phosphatdehydrogenase or 6-phosphogluconate-dehydrogenase

    DEFF Research Database (Denmark)

    Matzen, L E; Kristensen, S R; Kvetny, J

    1991-01-01

    The T3 modulation of nuclear T3 binding (NBT3), the T3 effect on cell growth, and the T3 and insulin effects on malic enzyme (ME), glucose-6-phosphat-dehydrogenase (G6PD) and 6-phosphogluconat-dehydrogenase (G6PD) were studied in a human hepatocyte cell-line (Chang-liver). T3 was bound to a high...

  18. Comparison of only T3 and T3-T4 sympathectomy for axillary hyperhidrosis regarding treatment effect and compensatory sweating.

    Science.gov (United States)

    Yuncu, Gökhan; Turk, Figen; Ozturk, Gökhan; Atinkaya, Cansel

    2013-08-01

    Patients diagnosed with axillary hyperhidrosis can face psychosocial issues that can ultimately hinder their quality of life both privately and socially. The routine treatment for axillary hyperhidrosis is T3-T4 sympathectomy, but compensatory sweating is a serious side effect that is commonly seen with this approach. This study was designed to evaluate whether a T3 sympathectomy was effective for the treatment of axillary hyperhidrosis and whether this treatment led to less compensatory sweating than T3-T4 sympathectomies among our 60-patient population. One hundred and twenty endoscopic thoracic sympathectomies were performed on 60 patients who had axillary hyperhidrosis. The sympathectomies were accomplished by means of a single-lumen endotracheal tube and a single port. The axillary hyperhidrosis patients were randomly divided into two groups with 17 patients in Group 1 undergoing T3-T4 sympathectomies and 43 in Group 2 undergoing only T3 sympathectomies. We analysed the data associated with the resolution of axillary hyperhidrosis, the degree of patient satisfaction with the surgical outcome and the quality of life in parallel with compensatory sweating after the procedure as reported by the patient and confirmed by the examiner. Moreover, the results were compared statistically. No statistically significant difference was observed between the groups based on age (P=0.56), gender (P=0.81), duration of the surgery (P=0.35) or postoperative satisfaction levels (P=0.45). However, the incidence and degree of compensatory sweating were lower in the T3 group than the T3-T4 group at the 1-year follow-up (P=0.008). T3 sympathectomy was as effective as T3-T4 sympathectomy for the treatment of axillary hyperhidrosis based on the patients' reported postoperative satisfaction, and the T3 group demonstrated lower compensatory sweating at the 1-year follow-up.

  19. Clinical Studies of Postoperative Arterial Infusion Chemotherapy in Patients with Pathologic T3 Esophageal Squamous Carcinoma

    Institute of Scientific and Technical Information of China (English)

    Baodong Liu; Zongjun Dong; Xiuyi Zhi; Qingsheng Xu

    2006-01-01

    OBJECTIVE To evaluate how arterial infusion chemotherapy after radical surgery influences long-term survival of patients with pathologic T3 (pT3) esophageal squamous carcinoma.METHODS We divided 190 patients with pathologic pT3 esophageal squamous carcinoma, confirmed by consecutive radical surgery, into an experimental group (surgery + intra-arterial infusion, 56 T3N0M0 and 52 T3N1M0 cases), and the remaining patients into a control group (surgery alone, 48 T3N0M0 and 34 T3N1M0 cases). The experimental group was sub-grouped into 56 cases (26 T3N0M0 and 30 T3N1M0 cases) receiving 1 or 2 periods of chemotherapy, while 52 cases (30 T3N0M0 and 22 T3N1M0 cases) underwent 3 or more than 3 periods of chemotherapy. We used one to seven courses of selected arterial infusion chemotherapy of cisplatin (80 mg/m2 of body-surface area) and fluorouracil (800 mg/m2) with or without epirubicin at 3~4 weeks post operation. The interval between each period was 3~4 weeks. All cases were followed-up for more than 5 years. Survival rates were calculated by the Kaplan-Meier methods and survival differences between patients with and without selected arterial infusion chemotherapy were compared with the Log-rank test. Prognostic variables were entered into a Cox regression analysis model controlling for age, site, lymph node status, and treatment received.RESULTS The overall survival rates were not significantly different between the experimental group and the control group, but there was better survival for patients who received 3 or more than 3 courses of chemotherapy. Lymph node status (N) was an important factor in the prognosis.CONCLUSION Trans-catheter arterial infusion chemotherapy is a safe and effective method of therapy. Postoperative selective arterial infusion chemotherapy can improve the survival rate in patients with esophageal squamous carcinoma who were previously treated by radical surgery.However, this modality of therapy needs further investigation.

  20. Triple3 Redundant Spacecraft Subsystems (T3RSS) Project

    Data.gov (United States)

    National Aeronautics and Space Administration — T3RSS is the system engineer's tool that allows a systematic approach to ensuring that even if one or more failures occur in a single component or subsystem, then...

  1. Phosphatidylcholine induces apoptosis of 3T3-L1 adipocytes

    Directory of Open Access Journals (Sweden)

    Li Hailan

    2011-12-01

    Full Text Available Abstract Background Phosphatidylcholine (PPC formulation is used for lipolytic injection, even though its mechanism of action is not well understood. Methods The viability of 3T3-L1 pre-adipocytes and differentiated 3T3-L1 cells was measured after treatment of PPC alone, its vehicle sodium deoxycholate (SD, and a PPC formulation. Western blot analysis was performed to examine PPC-induced signaling pathways. Results PPC, SD, and PPC formulation significantly decreased 3T3-L1 cell viability in a concentration-dependent manner. PPC alone was not cytotoxic to CCD-25Sk human fibroblasts at concentrations Conclusions PPC results in apoptosis of 3T3-L1 cells.

  2. T3: an international project for teacher training

    Directory of Open Access Journals (Sweden)

    Stefania Manca

    1996-01-01

    Full Text Available Presentation of T3, a project funded by the European Commission that focuses on providing distance training for teachers of mathematics, science, environmental education and foreign languages.

  3. Deiodination and conjugation of T3 and other iodothyronines

    OpenAIRE

    Eelkman Rooda, S.J.

    1988-01-01

    textabstractIodine enters the thyroid follicular cells as inorganic iodide and is incorporated through a series of metabolic steps into the main thyroid hormones thyroxine (T4 ) and 3,3',5-triiodothyronine (T3 ). Consecutive steps in this metabolic sequence are: 1) active transport of iodide by the thyroid follicular cell; 2) iodination of tyrosyl residues of thyroglobulin at the apical membrane; 3) coupling of mono- and diiodotyrosine molecules within thyroglobulin to form T4 and T 3 ; 4) pr...

  4. Defending plasma T3 is a biological priority.

    Science.gov (United States)

    Abdalla, Sherine M; Bianco, Antonio C

    2014-11-01

    Triiodothyronine (T3), the active form of thyroid hormone is produced predominantly outside the thyroid parenchyma secondary to peripheral tissue deiodination of thyroxine (T4), with thyroid. In healthy individuals, plasma T3 is regulated by the negative feedback loop of the hypothalamus-pituitary-thyroid axis and by homoeostatic changes in deiodinase expression. Therefore, with the exception of a minimal circadian rhythmicity, serum T3 levels are stable over long periods of time. Studies in rodents indicate that different levels of genetic disruption of the feedback mechanism and deiodinase system are met with increase in serum T4 and thyroid-stimulating hormone (TSH) levels, while serum T3 levels remain stable. These findings have focused attention on serum T3 levels in patients with thyroid disease, with important clinical implications affecting therapeutic goals and choice of therapy for patients with hypothyroidism. Although monotherapy with levothyroxine is the standard of care for hypothyroidism, not all patients normalize serum T3 levels with many advocating for combination therapy with levothyroxine and liothyronine. The latter could be relevant for a significant number of patients that remain symptomatic on monotherapy with levothyroxine, despite normalization of serum TSH levels. © 2014 John Wiley & Sons Ltd.

  5. Prolonged induction activates Cebpα independent adipogenesis in NIH/3T3 cells.

    Directory of Open Access Journals (Sweden)

    Hsiao-Yun Shao

    Full Text Available 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.

  6. Mediator subunit MED1 is a T3-dependent and T3-independent coactivator on the thyrotropin β gene promoter

    Energy Technology Data Exchange (ETDEWEB)

    Matsui, Keiji; Oda, Kasumi; Mizuta, Shumpei; Ishino, Ruri; Urahama, Norinaga; Hasegawa, Natsumi [Laboratory of Hematology, Division of Medical Biophysics, Kobe University Graduate School of Health Sciences, 7-10-2 Tomogaoka, Suma-ku, Kobe 654-0142 (Japan); Roeder, Robert G. [Laboratory of Biochemistry and Molecular Biology, The Rockefeller University, 1230 York Avenue, New York, NY 10065 (United States); Ito, Mitsuhiro, E-mail: itomi@med.kobe-u.ac.jp [Laboratory of Hematology, Division of Medical Biophysics, Kobe University Graduate School of Health Sciences, 7-10-2 Tomogaoka, Suma-ku, Kobe 654-0142 (Japan); Laboratory of Biochemistry and Molecular Biology, The Rockefeller University, 1230 York Avenue, New York, NY 10065 (United States); Department of Family and Community Medicine, Kobe University Graduate School of Medicine, 7-5-1 Kusunoki-cho, Chuo-ku, Kobe 654-0142 (Japan); Consolidated Research Institute for Advanced Science and Medical Care, Waseda University, 3-4-1 Okubo, Shinjuku-ku, Tokyo 159-8555 (Japan)

    2013-10-11

    Highlights: •MED1 is a bona fide T3-dependent coactivator on TSHB promoter. •Mice with LxxLL-mutant MED1 have attenuated TSHβ mRNA and thyroid hormone levels. •MED1 activates TSHB promoter T3-dependently in cultured cells. •T3-dependent MED1 action is enhanced when SRC1/SRC2 or HDAC2 is downregulated. •MED1 is also a T3-independent GATA2/Pit1 coactivator on TSHB promoter. -- Abstract: The MED1 subunit of the Mediator transcriptional coregulator complex is a nuclear receptor-specific coactivator. A negative feedback mechanism of thyroid-stimulating hormone (TSH, or thyrotropin) expression in the thyrotroph in the presence of triiodothyronine (T3) is employed by liganded thyroid hormone receptor β (TRβ) on the TSHβ gene promoter, where conventional histone-modifying coactivators act as corepressors. We now provide evidence that MED1 is a ligand-dependent positive cofactor on this promoter. TSHβ gene transcription was attenuated in MED1 mutant mice in which the nuclear receptor-binding ability of MED1 was specifically disrupted. MED1 stimulated GATA2- and Pit1-mediated TSHβ gene promoter activity in a ligand-independent manner in cultured cells. MED1 also stimulated transcription from the TSHβ gene promoter in a T3-dependent manner. The transcription was further enhanced when the T3-dependent corepressors SRC1, SRC2, and HDAC2 were downregulated. Hence, MED1 is a T3-dependent and -independent coactivator on the TSHβ gene promoter.

  7. ExsE Is a Negative Regulator for T3SS Gene Expression in Vibrio alginolyticus

    Science.gov (United States)

    Liu, Jinxin; Lu, Shao-Yeh; Orfe, Lisa H.; Ren, Chun-Hua; Hu, Chao-Qun; Call, Douglas R.; Avillan, Johannetsy J.; Zhao, Zhe

    2016-01-01

    Type III secretion systems (T3SSs) contribute to microbial pathogenesis of Vibrio species, but the regulatory mechanisms are complex. We determined if the classic ExsACDE protein-protein regulatory model from Pseudomonas aeruginosa applies to Vibrio alginolyticus. Deletion mutants in V. alginolyticus demonstrated that, as expected, the T3SS is positively regulated by ExsA and ExsC and negatively regulated by ExsD and ExsE. Interestingly, deletion of exsE enhanced the ability of V. alginolyticus to induce host-cell death while cytotoxicity was inhibited by in trans complementation of this gene in a wild-type strain, a result that differs from a similar experiment with Vibrio parahaemolyticus ExsE. We further showed that ExsE is a secreted protein that does not contribute to adhesion to Fathead minnow epithelial cells. An in vitro co-immunoprecipitation assay confirmed that ExsE binds to ExsC to exert negative regulatory effect on T3SS genes. T3SS in V. alginolyticus can be activated in the absence of physical contact with host cells and a separate regulatory pathway appears to contribute to the regulation of ExsA. Consequently, like ExsE from P. aeruginosa, ExsE is a negative regulator for T3SS gene expression in V. alginolyticus. Unlike the V. parahaemolyticus orthologue, however, deletion of exsE from V. alginolyticus enhanced in vitro cytotoxicity. PMID:27999769

  8. Isoproterenol Increases Uncoupling, Glycolysis, and Markers of Beiging in Mature 3T3-L1 Adipocytes.

    Science.gov (United States)

    Miller, Colette N; Yang, Jeong-Yeh; England, Emily; Yin, Amelia; Baile, Clifton A; Rayalam, Srujana

    2015-01-01

    Beta-adrenergic activation stimulates uncoupling protein 1 (UCP1), enhancing metabolic rate. In vitro, most work has studied brown adipocytes, however, few have investigated more established adipocyte lines such as the murine 3T3-L1 line. To assess the effect of beta-adrenergic activation, mature 3T3-L1s were treated for 6 or 48 hours with or without isoproterenol (10 and 100 μM) following standard differentiation supplemented with thyroid hormone (T3; 1 nM). The highest dose of isoproterenol increased lipid content following 48 hours of treatment. This concentration enhanced UCP1 mRNA and protein expression. The increase in UCP1 following 48 hours of isoproterenol increased oxygen consumption rate. Further, coupling efficiency of the electron transport chain was disturbed and an enhancement of glycolytic rate was measured alongside this, indicating an attempt to meet the energy demands of the cell. Lastly, markers of beige adipocytes (protein content of CD137 and gene transcript of CITED1) were also found to be upregulated at 48 hours of isoproterenol treatment. This data indicates that mature 3T3-L1 adipocytes are responsive to isoproterenol and induce UCP1 expression and activity. Further, this finding provides a model for further pharmaceutical and nutraceutical investigation of UCP1 in 3T3-L1s.

  9. Isoproterenol Increases Uncoupling, Glycolysis, and Markers of Beiging in Mature 3T3-L1 Adipocytes.

    Directory of Open Access Journals (Sweden)

    Colette N Miller

    Full Text Available Beta-adrenergic activation stimulates uncoupling protein 1 (UCP1, enhancing metabolic rate. In vitro, most work has studied brown adipocytes, however, few have investigated more established adipocyte lines such as the murine 3T3-L1 line. To assess the effect of beta-adrenergic activation, mature 3T3-L1s were treated for 6 or 48 hours with or without isoproterenol (10 and 100 μM following standard differentiation supplemented with thyroid hormone (T3; 1 nM. The highest dose of isoproterenol increased lipid content following 48 hours of treatment. This concentration enhanced UCP1 mRNA and protein expression. The increase in UCP1 following 48 hours of isoproterenol increased oxygen consumption rate. Further, coupling efficiency of the electron transport chain was disturbed and an enhancement of glycolytic rate was measured alongside this, indicating an attempt to meet the energy demands of the cell. Lastly, markers of beige adipocytes (protein content of CD137 and gene transcript of CITED1 were also found to be upregulated at 48 hours of isoproterenol treatment. This data indicates that mature 3T3-L1 adipocytes are responsive to isoproterenol and induce UCP1 expression and activity. Further, this finding provides a model for further pharmaceutical and nutraceutical investigation of UCP1 in 3T3-L1s.

  10. Teachers Teaching Teachers (T3) [TM]. Vol. 2 No. 7

    Science.gov (United States)

    Richardson, Joan, Ed.

    2007-01-01

    This issue of "Teachers Teaching Teachers" ("T3") focuses on coaches' role in the professional development of teachers. It contains the following articles: (1) An Excerpt from "Taking the Lead" (Joellen Killion and Cindy Harrison); (2) Be Like a Virus and Connect (Bill Ferriter); (3) No. 1 Resource Has a Human Face (Joellen Killion); (4) With This…

  11. Deiodination and conjugation of T3 and other iodothyronines

    NARCIS (Netherlands)

    S.J. Eelkman Rooda

    1988-01-01

    textabstractIodine enters the thyroid follicular cells as inorganic iodide and is incorporated through a series of metabolic steps into the main thyroid hormones thyroxine (T4 ) and 3,3',5-triiodothyronine (T3 ). Consecutive steps in this metabolic sequence are: 1) active transport of iodide by

  12. Reply to ``Comment on `T3 specific-heat anomaly in network solids' ''

    Science.gov (United States)

    Phillips, J. C.

    1987-02-01

    My paper asserted that a wide range of data supports a structural model of melt-quenched vitreous silica based on defective cristobalite microcrystallites. These defects may contribute to the apparent ``excess T3'' term in the specific heat. This model is supported by recent data on porous vitreous silica prepared by the sol-gel method. The older model, based on analogies with the vibrational spectrum of crystalline Ge, has become less plausible in the overall context of presently available data.

  13. Isobaric multiplet mass equation in the A =31 ,T =3 /2 quartets

    Science.gov (United States)

    Bennett, M. B.; Wrede, C.; Brown, B. A.; Liddick, S. N.; Pérez-Loureiro, D.; Bardayan, D. W.; Chen, A. A.; Chipps, K. A.; Fry, C.; Glassman, B. E.; Langer, C.; Larson, N. R.; McNeice, E. I.; Meisel, Z.; Ong, W.; O'Malley, P. D.; Pain, S. D.; Prokop, C. J.; Schwartz, S. B.; Suchyta, S.; Thompson, P.; Walters, M.; Xu, X.

    2016-06-01

    Background: The observed mass excesses of analog nuclear states with the same mass number A and isospin T can be used to test the isobaric multiplet mass equation (IMME), which has, in most cases, been validated to a high degree of precision. A recent measurement [Kankainen et al., Phys. Rev. C 93, 041304(R) (2016), 10.1103/PhysRevC.93.041304] of the ground-state mass of 31Cl led to a substantial breakdown of the IMME for the lowest A =31 ,T =3 /2 quartet. The second-lowest A =31 ,T =3 /2 quartet is not complete, due to uncertainties associated with the identity of the 31S member state. Purpose: Our goal is to populate the two lowest T =3 /2 states in 31S and use the data to investigate the influence of isospin mixing on tests of the IMME in the two lowest A =31 ,T =3 /2 quartets. Methods: Using a fast 31Cl beam implanted into a plastic scintillator and a high-purity Ge γ -ray detection array, γ rays from the 31Cl(β γ )31S sequence were measured. Shell-model calculations using USDB and the recently-developed USDE interactions were performed for comparison. Results: Isospin mixing between the 31S isobaric analog state (IAS) at 6279.0(6) keV and a nearby state at 6390.2(7) keV was observed. The second T =3 /2 state in 31S was observed at Ex=7050.0 (8 ) keV. Calculations using both USDB and USDE predict a triplet of isospin-mixed states, including the lowest T =3 /2 state in 31P, mirroring the observed mixing in 31S, and two isospin-mixed triplets including the second-lowest T =3 /2 states in both 31S and 31P. Conclusions: Isospin mixing in 31S does not by itself explain the IMME breakdown in the lowest quartet, but it likely points to similar isospin mixing in the mirror nucleus 31P, which would result in a perturbation of the 31P IAS energy. USDB and USDE calculations both predict candidate 31P states responsible for the mixing in the energy region slightly above Ex=6400 keV. The second quartet has been completed thanks to the identification of the second 31S T

  14. [Controversy over treatment options in clinical T3 prostate cancer].

    Science.gov (United States)

    Nishimura, Kazuo; Yamamoto, Yoshiyuki; Nakayama, Masashi; Kakimoto, Ken-ichi

    2011-12-01

    Clinical stage T3 prostate cancer is a locally advanced disease at risk of having micrometastasis.Since clinical T staging is potentially inaccurate, PSA and Gleason score are also added at biopsy for risk stratification. The recurrence rate after radical prostatectomy alone or radiotherapy alone is generally high. Therefore, multimodal treatment is required. Recent multiple randomized trials have shown survival benefits of radiotherapy combined with long-term hormonal therapy. Dose escalation utilizing intensity modulated radiotherapy (IMRT) or brachytherapy has the potential benefit of exerting local control. Since previous trials used conventional external radiotherapy, the optimal duration of hormonal therapy combined with dose escalated radiotherapy remains unknown. On the other hand, some patients can be cured by radical prostatectomy alone, and approximately half of patients with adverse features after surgery can be rescued by adjuvant radiotherapy. Primary hormonal therapy is used widely and has shown favorable results in patients with T3 prostate cancer, particularly in Japan. Based on the life expectancy and comorbidity of an individual patient, hormonal therapy can be chosen as a primary treatment. There are controversies over treatment options such as surgery, radiotherapy, and hormonal therapy in clinical T3 prostate cancer. The clinical results of these treatments are reviewed and several unresolved issues are addressed here.

  15. Hormone (ACTH, T3) content of immunophenotyped lymphocyte subpopulations.

    Science.gov (United States)

    Pállinger, Éva; Kiss, Gergely Attila; Csaba, György

    2016-12-01

    Cells of the immune system synthesize, store, and secrete polypeptide and amino acid type hormones, which also influence their functions, having receptors for different hormones. In the present experiment immunophenotyped immune cells isolated from bone marrow, thymus, and peritoneal fluid of mice were used for demonstrating the adrenocorticotropic hormone (ACTH) and triiodothyronine (T3) hormone production of differentiating immune cells. Both hormones were found in each cell type, and in each maturation state, which means that all cells are participating in the hormonal function of the immune system. The lineage-independent presence of ACTH and T3 in differentiating hematopoietic cells denotes that their expression ubiquitous during lymphocyte development. Higher ACTH and T3 content of B cells shows that these cells are the most hormonally active and suggests that the hormones may have an autocrine regulatory role in B cell development. Developing T cells showed heterogeneous hormone production which was associated with their maturation state. Differences in the hormone contents of immune cells isolated from different organs indicate that their hormone production is defined by their differentiation or maturation state, however, possibly also by the local microenvironment.

  16. Growth stimulation of 3T3 fibroblasts by Cystatin

    Energy Technology Data Exchange (ETDEWEB)

    Quan Sun (Michigan State Univ., East Lansing (United States) Beijing Medical Univ. (China))

    1989-01-01

    Treatment of cultures of mouse 3T3 fibroblasts with Cystatin C, a thiol-proteinase inhibitor isolated from chicken egg white, resulted in an enhanced rate of cell proliferation. This stimulation was demonstrated using two independent assay systems: (a) assessment of total cell number and (b) measurement of ({sup 3}H)thymidine incorporated into acid-precipitable DNA. In both assays, the dose-response curves of Cystatin stimulation showed a rising function that plateaued at a concentration of {approximately}120 {mu}g/ml. The addition of Cystatin to cultures of Kirsten murine sarcoma virus-transformed 3T3 cells also enhanced DNA synthesis in these target cells. Control experiments showed that the presence of Cystatin did not alter the level of binding of radioactively labeled epidermal growth factor and platelet derived growth factor to 3T3 cells. These results argue against the possibility that the observed growth stimulation by Cystatin was due to growth factor contamination of the Cystatin preparation.

  17. Temperature distribution study during the friction stir welding process of Al2024-T3 aluminum alloy

    Science.gov (United States)

    Yau, Y. H.; Hussain, A.; Lalwani, R. K.; Chan, H. K.; Hakimi, N.

    2013-08-01

    Heat flux characteristics are critical to good quality welding obtained in the important engineering alloy Al2024-T3 by the friction stir welding (FSW) process. In the present study, thermocouples in three different configurations were affixed on the welding samples to measure the temperatures: in the first configuration, four thermocouples were placed at equivalent positions along one side of the welding direction; the second configuration involved two equivalent thermocouple locations on either side of the welding path; while the third configuration had all the thermocouples on one side of the layout but with unequal gaps from the welding line. A three-dimensional, non-linear ANSYS computational model, based on an approach applied to Al2024-T3 for the first time, was used to simulate the welding temperature profiles obtained experimentally. The experimental thermal profiles on the whole were found to be in agreement with those calculated by the ANSYS model. The broad agreement between the two kinds of profiles validates the basis for derivation of the simulation model and provides an approach for the FSW simulation in Al2024-T3 and is potentially more useful than models derived previously.

  18. T3PS: Tool for Parallel Processing in Parameter Scans

    CERN Document Server

    Maurer, Vinzenz

    2015-01-01

    T3PS is a program that can be used to quickly design and perform parameter scans while easily taking advantage of the multi-core architecture of current processors. It takes an easy to read and write parameter scan definition file format as input. Based on the parameter ranges and other options contained therein, it distributes the calculation of the parameter space over multiple processes and possibly computers. The derived data is saved in a plain text file format readable by most plotting software. The supported scanning strategies include: grid scan, random scan, Markov Chain Monte Carlo, numerical optimization. Several example parameter scans are shown and compared with results in the literature.

  19. Aspartame downregulates 3T3-L1 differentiation.

    Science.gov (United States)

    Pandurangan, Muthuraman; Park, Jeongeun; Kim, Eunjung

    2014-10-01

    Aspartame is an artificial sweetener used as an alternate for sugar in several foods and beverages. Since aspartame is 200 times sweeter than traditional sugar, it can give the same level of sweetness with less substance, which leads to lower-calorie food intake. There are reports that consumption of aspartame-containing products can help obese people lose weight. However, the potential role of aspartame in obesity is not clear. The present study investigated whether aspartame suppresses 3T3-L1 differentiation, by downregulating phosphorylated peroxisome proliferator-activated receptor γ (p-PPARγ), peroxisome proliferator-activated receptor γ (PPARγ), fatty acid-binding protein 4 (FABP4), CCAAT/enhancer-binding protein α (C/EBPα), and sterol regulatory element-binding protein 1 (SREBP1), which are critical for adipogenesis. The 3T3-L1 adipocytes were cultured and differentiated for 6 d in the absence and presence of 10 μg/ml of aspartame. Aspartame reduced lipid accumulation in differentiated adipocytes as evidenced by Oil Red O staining. qRT-PCR analysis showed that the PPARγ, FABP4, and C/EBPα mRNA expression was significantly reduced in the aspartame-treated adipocytes. Western blot analysis showed that the induction of p-PPARγ, PPARγ, SREBP1, and adipsin was markedly reduced in the aspartame-treated adipocytes. Taken together, these data suggest that aspartame may be a potent substance to alter adipocyte differentiation and control obesity.

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

    Science.gov (United States)

    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.

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

    Energy Technology Data Exchange (ETDEWEB)

    Lai, Jing [The First Affiliated Hospital, Jinan University, Guangzhou, Guangdong (China); Liu, Gexiu [Institute of Hematology, School of Medicine, Jinan University, Guangzhou, Guangdong (China); Yan, Guoyao [The First Affiliated Hospital, Jinan University, Guangzhou, Guangdong (China); He, Dongmei [Institute of Hematology, School of Medicine, Jinan University, Guangzhou, Guangdong (China); Zhou, Ying [The First Affiliated Hospital, Jinan University, Guangzhou, Guangdong (China); Chen, Shengting, E-mail: shengtingchen@sina.cn [The First Affiliated Hospital, Jinan University, Guangzhou, Guangdong (China)

    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.

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

    Science.gov (United States)

    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.

  3. Tensile and shear failure mechanisms of 2024-T3 aluminum alloy%2024-T3铝合金拉伸及剪切断裂行为

    Institute of Scientific and Technical Information of China (English)

    姜薇; 李亚智; 苏杰; 束一秀

    2015-01-01

    2024铝合金材料在拉伸和扭转载荷作用下表现出截然不同的失效机理。结合试验和数值方法,研究了应力状态对2024⁃T3铝合金韧性断裂行为的影响规律。首先,对圆棒和薄壁圆筒试验件分别进行了拉伸和扭转试验,从断面形貌以及断裂应变与应力状态间关系两个方面,考察了应力状态对2024⁃T3铝合金断裂机理的影响规律。然后,基于Gurson理论在商业有限元软件ABAQUS中开发了同时适用于拉伸和剪切断裂模式的细观损伤本构,对2024⁃T3铝合金的弹塑性响应和裂纹扩展路径进行了数值分析。与试验结果对比研究表明,本文发展的细观损伤本构能够较好预测延性金属材料在多种应力状态下的损伤破坏过程。%Under tensile and shear loading conditions, 2024 aluminum alloy exhibits two types of distinctive ductile rupture mechanisms. The growth and internal necking of voids governs the rupture mechanism in tension dominated loading mode, while the internal shearing in the ligaments between voids dominants for shear conditions. To investigate the influence of stress states on the material ductility of 2024⁃T3 aluminum alloy, tensile experiments of a smooth round bar and three notched round bars with different notch root radii as well as a pure torsion experiment were performed. Based on the modification of Gurson model by Nahshon and Hutchinson, a void⁃based meso⁃damage constitutive relationship which can deal with both tensile and shear problems was developed and implemented in commercial software ABAQUS. The tensile and shear fracture behaviors of 2024⁃T3 aluminum alloy including the load⁃displacement response and crack propagation path were analyzed using the proposed approach and compared with experi⁃mental data. It is shown that the proposed approach can be used to predict the failure of ductile materials under complex loading conditions.

  4. F1t3 RECEPTOR EXPRESSION ON THE SURFACE OF MALIGNANT HEMATOPOIETIC CELLS AND RESPONSES TO F1t3 LIGAND STIMULATION

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    Objective: To investigate the F1t3 receptor expression on the surface of malignant hematopoietic cells, the effect of TNFa and dexamethasone (DXM) on its expression and the responses of those cells to recombinant human F1t3 ligand (rhFL). Methods: Eighteen malignant hematopoietic cell lines were determined for the F1t3 receptor expression by flow cytometric analysis. The effect of rhFL on the proliferation of malignant hematopoietic cells in vitro was measured using MTT assay. Results: The expressions of F1t3 receptor on the surface of Raji, Daudi, HL-60, 8266 and XG-6 cells were detected by flow cytometric analysis. Following incubation with 20 ng/ml TNFa for 24h, the number of F1t3 receptor positive cells decreased in Raji and 8266, increased in HL-60 and XG-6, and no difference in Daudi cells. After incubation with 10-6 mol/L DXM for 24h, the number of F1t3 receptor positive cells decreased in all the 5 F1t3 receptor positive cell lines. rhFL stimulated the proliferation of HL-60 and Raji cells. Conclusion: For most of the malignant hematopoietic cells, there was neither the expression of F1t3 receptor nor the response to rhFL. DXM may be useful to reduce the effect of FL on the proliferation of some F1t3 receptor positive malignant hematopoietic cells in vitro and in vivo.

  5. Protection of 2024-T3 aluminium alloy by corrosion resistant phytic acid conversion coating

    Science.gov (United States)

    Shi, Hongwei; Han, En-Hou; Liu, Fuchun; Kallip, Silvar

    2013-09-01

    The corrosion protection properties of environmentally friendly phytic acid conversion coatings were studied on 2024-T3 aluminium alloy. The films were prepared under acidic conditions with various pH values and characterised by SEM, EDS, ATR-FTIR and electrochemical techniques. The results indicate that the conversion coatings obtained by immersing the alloy in phytic acid solutions at pH from 3 to 5.5 provide excellent corrosion resistance. ATR-FTIR confirms that the film is formed by deposition of reaction products between Al3+ and phosphate groups in phytic acid molecules. The conformation models of the deposition film are proposed.

  6. Message passing performance of Intel Paragon, IBM SP1 and Cray T3D using PVM

    Energy Technology Data Exchange (ETDEWEB)

    Manke, J.W.; Patterson, J.C. [Boeing Computer Services, Seattle, WA (United States)

    1995-12-01

    For distributed applications on MPP machinse, message passing performance is a critical factor in the overall speed and scalability of the application. This is particularly true when all-to-all communication between nodes is required. In this study we measured the message passing performance of our PVM implementation of an all-to-all communication method called recursive doubling. The measurements were done on an Intel Paragon, IBM SP1 and CRAY T3D. Using a model of message passing times for recursive doubling, we developed several measures that can be used to compare the message passing performance of MPP machines.

  7. Evaluation of chylomicron effect on ASP production in 3T3-L1 adipocytes.

    Science.gov (United States)

    Gao, Ying; Gauvreau, Danny; Cui, Wei; Lapointe, Marc; Paglialunga, Sabina; Cianflone, Katherine

    2011-02-01

    In the past few years, there has been increasing interest in the production and physiological role of acylation-stimulating protein (ASP), identical to C3adesArg, a product of the alternative complement pathway generated through C3 cleavage. Recent studies in C3 (-/-) mice that are ASP deficient have demonstrated a role for ASP in postprandial triglyceride clearance and fat storage. The aim of the present study was to establish a cell model and sensitive ELISA assay for the evaluation of ASP production using 3T3-L1 adipocytes. 3T3-L1 preadipocytes were differentiated into adipocytes, then cultured in different media such as serum-free (SF), Dulbecco's modified Eagle's medium (DMEM)/F12 + 10% fetal calf serum (FBS), and at varying concentrations of chylomicrons and insulin + chylomicrons up to 48 h. ASP production in SF and DMEM/F12 + 10% FBS was compared. Chylomicrons stimulated ASP production in a concentration- and time-dependent manner. By contrast, chylomicron treatment had no effect on the production of C3, the precursor protein of ASP, which was constant over 48 h. Addition of insulin (100 nM) to a low-dose of chylomicrons (100 µg TG/ml) significantly increased ASP production compared with chylomicrons alone at 48 h (P ASP to C3 remained constant, indicating no change in the ratio of C3 cleaved to generate ASP. This study demonstrated that 3T3-L1 adipocyte is a useful model for the evaluation of C3 secretion and ASP production by using a sensitive mouse-specific ELISA assay. The stimulation of ASP production with chylomicrons demonstrates a physiologically relevant response, and provides a strategy for further studies on ASP production and function.

  8. Effect of Black Soybean Koji Extract on Glucose Utilization and Adipocyte Differentiation in 3T3-L1 Cells

    Directory of Open Access Journals (Sweden)

    Chi-Chang Huang

    2014-05-01

    Full Text Available Adipocyte differentiation and the extent of subsequent fat accumulation are closely related to the occurrence and progression of diseases such as insulin resistance and obesity. Black soybean koji (BSK is produced by the fermentation of black soybean with Aspergilllus awamori. Previous study indicated that BSK extract has antioxidative and multifunctional bioactivities, however, the role of BSK in the regulation of energy metabolism is still unclear. We aimed to investigate the effect of glucose utilization on insulin-resistant 3T3-L1 preadipocytes and adipogenesis-related protein expression in differentiated adipocytes with BSK treatment. Cytoxicity assay revealed that BSK did not adversely affect cell viability at levels up to 200 µg/mL. The potential for glucose utilization was increased by increased glucose transporter 1 (GLUT1, GLUT4 and protein kinase B (AKT protein expression in insulin-resistant 3T3-L1 cells in response to BSK treatment. Simultaneously, BSK inhibited lipid droplet accumulation in differentiated 3T3-L1 cells. The inhibitory effect of adipogenesis was associated with downregulated peroxisome proliferator-activated receptor g (PPARγ level and upregulated Acrp30 protein expression. Our results suggest that BSK extract could improve glucose uptake by modulating GLUT1 and GLUT4 expression in a 3T3-L1 insulin-resistance cell model. In addition, BSK suppressed differentiation and lipid accumulation in mature 3T3-L1 adipocytes, which may suggest its potential for food supplementation to prevent obesity and related metabolic abnormalities.

  9. Berberine Alleviates Olanzapine-Induced Adipogenesis via the AMPKα–SREBP Pathway in 3T3-L1 Cells

    OpenAIRE

    Yanjie Li; Xiaomin Zhao; Xiyu Feng; Xuemei Liu; Chao Deng; Chang-Hua Hu

    2016-01-01

    The aim of this study was to investigate the mechanisms underlying the inhibitory effects of berberine (BBR) on olanzapine (OLZ)-induced adipogenesis in a well-replicated 3T3-L1 cell model. Oil-Red-O (ORO) staining showed that BBR significantly decreased OLZ-induced adipogenesis. Co-treatment with OLZ and BBR decreased the accumulation of triglyceride (TG) and total cholesterol (TC) by 55.58% ± 3.65% and 49.84% ± 8.31%, respectively, in 3T3-L1 adipocytes accompanied by reduced expression of S...

  10. Comparison of only T3 and T3–T4 sympathectomy for axillary hyperhidrosis regarding treatment effect and compensatory sweating

    Science.gov (United States)

    Yuncu, Gökhan; Turk, Figen; Ozturk, Gökhan; Atinkaya, Cansel

    2013-01-01

    OBJECTIVES Patients diagnosed with axillary hyperhidrosis can face psychosocial issues that can ultimately hinder their quality of life both privately and socially. The routine treatment for axillary hyperhidrosis is T3–T4 sympathectomy, but compensatory sweating is a serious side effect that is commonly seen with this approach. This study was designed to evaluate whether a T3 sympathectomy was effective for the treatment of axillary hyperhidrosis and whether this treatment led to less compensatory sweating than T3–T4 sympathectomies among our 60-patient population. METHODS One hundred and twenty endoscopic thoracic sympathectomies were performed on 60 patients who had axillary hyperhidrosis. The sympathectomies were accomplished by means of a single-lumen endotracheal tube and a single port. The axillary hyperhidrosis patients were randomly divided into two groups with 17 patients in Group 1 undergoing T3–T4 sympathectomies and 43 in Group 2 undergoing only T3 sympathectomies. We analysed the data associated with the resolution of axillary hyperhidrosis, the degree of patient satisfaction with the surgical outcome and the quality of life in parallel with compensatory sweating after the procedure as reported by the patient and confirmed by the examiner. Moreover, the results were compared statistically. RESULTS No statistically significant difference was observed between the groups based on age (P = 0.56), gender (P = 0.81), duration of the surgery (P = 0.35) or postoperative satisfaction levels (P = 0.45). However, the incidence and degree of compensatory sweating were lower in the T3 group than the T3–T4 group at the 1-year follow-up (P = 0.008). CONCLUSIONS T3 sympathectomy was as effective as T3–T4 sympathectomy for the treatment of axillary hyperhidrosis based on the patients’ reported postoperative satisfaction, and the T3 group demonstrated lower compensatory sweating at the 1-year follow-up. PMID:23644731

  11. Early life trauma is associated with decreased peripheral levels of thyroid-hormone T3 in adolescents.

    Science.gov (United States)

    Machado, T D; Salum, G A; Bosa, V L; Goldani, M Z; Meaney, M J; Agranonik, M; Manfro, G G; Silveira, P P

    2015-12-01

    An adverse early life environment can induce changes on behavioral and metabolic responses later in life. Recent studies in rats showed that the quality of maternal care as measured by high levels of pup licking and grooming (LG) was associated with changes in the relationship between the precursor thyroid-hormone T4 and the more active T3. Here we investigated if early exposure to childhood abuse is associated with thyroid-hormone levels in human adolescents. Given the empirical evidence from animal models showing that good maternal care was associated with increased conversion of T4 to T3, we hypothesized that early adversity would be associated with a decreased peripheral conversion of T4 to T3. A sample of 80 adolescents (10-18 years) participated in this study. We used the Childhood Trauma Questionnaire to investigate early life stress. We calculate the body mass index (BMI) assessing weight and height and sexual maturation stage was determined by self-assessment. Blood samples were collected to measure T3 and T4 levels. ANCOVA were used to evaluate the influence of the Physical Abuse domain of the Childhood Trauma Questionnaire as the early life stress variable in T3 and T4 separately, adjusted for potential confounders such as pubertal status, gender, socioeconomic status and BMI. Early life trauma was associated with reduced T3 levels in adolescents, when adjusted for potential confounders (p=0.013), but not with peripheral T4 levels (p=0.625). We extended findings from animal models showing that adverse early experience persistently impacts on the individual's responses to stress, which is marked by an abnormal metabolism of thyroid hormones. Further studies are needed to further investigate the nature of such associations.

  12. Effect of triiodothyronine( T3) on apoptosis of endothelial cells and interventional study with osteoprotegerin

    Institute of Scientific and Technical Information of China (English)

    王浩华

    2014-01-01

    Objective To explore the effect and mechanism of osteoprotegerin on apoptosis in human umbilical vein endothelial cells(HUVECs)induced by triiodothyronine(T3)with different concentrations.Methods HUVECs were cultured in control group(0 nmol/L T3),low-concentration group(0.1 nmol/L T3),physiological-concentration group(1 nmol/L T3),high-concentration group(10 nmol/L T3)with OPG for 48 h.The cell apoptosis was measured by cytometry and Hoechst 33258.The expressions of p-IκKβ,IκKβ,p-

  13. Long-term L-Triiodothyronine (T3) treatment in stable systolic heart failure patients

    DEFF Research Database (Denmark)

    Holmager, Pernille; Schmidt, Ulla; Mark, Peter

    2015-01-01

    during T3-treatment and neither did the neurohormonal profile. There were no side-effects in terms of cardiac arrhythmias and no change in resting heart rate. CONCLUSIONS: This study does not support the hypothesis that oral T3 treatment might be beneficial to patients with chronic, stable systolic HF......BACKGROUND: Chronic heart failure (HF) is characterized by reduced serum T3 levels and increased activity of the T3 degrading enzyme deiodinase D3. This may result in an intracellular composition of the cardiomyocyte mimicking that of hypothyroidism. Short-term T3-administration to systolic HF...... patients might be beneficial. QUESTION: Does long-term treatment with T3 have a beneficial effect on cardiac function and neurohormonal activation in chronic systolic HF patients with serum T3 levels below 1·6 nmol/l? DESIGN: A randomized, double-blind, cross-over, placebo-controlled intervention study...

  14. Length quantization of DNA partially expelled from heads of a bacteriophage T3 mutant

    Energy Technology Data Exchange (ETDEWEB)

    Serwer, Philip, E-mail: serwer@uthscsa.edu [Department of Biochemistry, The University of Texas Health Science Center, 7703 Floyd Curl Drive, San Antonio, TX 78229-3900 (United States); Wright, Elena T. [Department of Biochemistry, The University of Texas Health Science Center, 7703 Floyd Curl Drive, San Antonio, TX 78229-3900 (United States); Liu, Zheng; Jiang, Wen [Markey Center for Structural Biology, Department of Biological Sciences, Purdue University, West Lafayette, IN 47907 (United States)

    2014-05-15

    DNA packaging of phages phi29, T3 and T7 sometimes produces incompletely packaged DNA with quantized lengths, based on gel electrophoretic band formation. We discover here a packaging ATPase-free, in vitro model for packaged DNA length quantization. We use directed evolution to isolate a five-site T3 point mutant that hyper-produces tail-free capsids with mature DNA (heads). Three tail gene mutations, but no head gene mutations, are present. A variable-length DNA segment leaks from some mutant heads, based on DNase I-protection assay and electron microscopy. The protected DNA segment has quantized lengths, based on restriction endonuclease analysis: six sharp bands of DNA missing 3.7–12.3% of the last end packaged. Native gel electrophoresis confirms quantized DNA expulsion and, after removal of external DNA, provides evidence that capsid radius is the quantization-ruler. Capsid-based DNA length quantization possibly evolved via selection for stalling that provides time for feedback control during DNA packaging and injection. - Graphical abstract: Highlights: • We implement directed evolution- and DNA-sequencing-based phage assembly genetics. • We purify stable, mutant phage heads with a partially leaked mature DNA molecule. • Native gels and DNase-protection show leaked DNA segments to have quantized lengths. • Native gels after DNase I-removal of leaked DNA reveal the capsids to vary in radius. • Thus, we hypothesize leaked DNA quantization via variably quantized capsid radius.

  15. Isobaric multiplet mass equation in the $A=31$ $T = 3/2$ quartets

    CERN Document Server

    Bennett, M B; Brown, B A; Liddick, S N; Pérez-Loureiro, D; Bardayan, D W; Chen, A A; Chipps, K A; Fry, C; Glassman, B E; Langer, C; Larson, N R; McNeice, E I; Meisel, Z; Ong, W; O'Malley, P D; Pain, S D; Prokop, C J; Schwartz, S B; Suchyta, S; Thompson, P; Walters, M; Xu, X

    2016-01-01

    The observed mass excesses of analog nuclear states with the same mass number $A$ and isospin $T$ can be used to test the isobaric multiplet mass equation (IMME), which has, in most cases, been validated to a high degree of precision. A recent measurement [Kankainen et al., Phys. Rev. C 93 041304(R) (2016)] of the ground-state mass of $^{31}$Cl led to a substantial breakdown of the IMME for the lowest $A = 31, T = 3/2$ quartet. The second-lowest $A = 31, T = 3/2$ quartet is not complete, due to uncertainties associated with the identity of the $^{31}$S member state. Using a fast $^{31}$Cl beam implanted into a plastic scintillator and a high-purity Ge $\\gamma$-ray detection array, $\\gamma$ rays from the $^{31}$Cl$(\\beta\\gamma)$$^{31}$S sequence were measured. Shell-model calculations using USDB and the recently-developed USDE interactions were performed for comparison. Isospin mixing between the $^{31}$S isobaric analog state (IAS) at 6279.0(6) keV and a nearby state at 6390.2(7) keV was observed. The second ...

  16. Berberine inhibits 3T3-L1 adipocyte differentiation through the PPARgamma pathway.

    Science.gov (United States)

    Huang, Cheng; Zhang, Yuebo; Gong, Zhenwei; Sheng, Xiaoyan; Li, Zongmeng; Zhang, Wei; Qin, Ying

    2006-09-22

    Berberine (BBR), a compound purified from Cortidis rhizoma, reduces serum cholesterol, triglycerides, and LDL-cholesterol of hypercholesterolemic patients and high fat diet fed animals, and increases hepatic LDLR mRNA and protein levels through a post-transcriptional mechanism. BBR also enhances the hypoglycemic action of insulin in diabetic animal models. Here, we show that BBR inhibits the differentiation of 3T3-L1 preadipocytes induced by DM and suppresses the mitotic clonal expansion of 3T3-L1 preadipocytes in a time- and dose-dependent manner. Gene expression analysis and Western blot analysis reveal that the BBR inhibits the mRNA and protein levels of adipogenesis related transcription factors PPARgamma and C/EBPalpha and their upstream regulator, C/EBPbeta. Reporter gene assays demonstrate that the full-length PPARgamma and alpha transcription activities are inhibited by BBR. Using real-time PCR, we have also found that the PPAR target genes that are involved in adipocyte differentiation, such as aP2, CD36, ACO, LPL, and other adipocyte markers, are suppressed by BBR. These studies suggest that BBR works on multiple molecular targets as an inhibitor of PPARgamma and alpha, and is a potential weight reducing, hypolipidemic, and hypoglycemic drug.

  17. Folate receptor {alpha} regulates cell proliferation in mouse gonadotroph {alpha}T3-1 cells

    Energy Technology Data Exchange (ETDEWEB)

    Yao, Congjun; Evans, Chheng-Orn [Department of Neurosurgery and Laboratory of Molecular Neurosurgery and Biotechnology, Emory University, School of Medicine, Atlanta, Georgia (United States); Stevens, Victoria L. [Epidemiology and Surveillance Research, American Cancer Society, Atlanta, Georgia (United States); Owens, Timothy R. [Emory University, School of Medicine, Atlanta, Georgia (United States); Oyesiku, Nelson M., E-mail: noyesik@emory.edu [Department of Neurosurgery and Laboratory of Molecular Neurosurgery and Biotechnology, Emory University, School of Medicine, Atlanta, Georgia (United States)

    2009-11-01

    We have previously found that the mRNA and protein levels of the folate receptor alpha (FR{alpha}) are uniquely over-expressed in clinically human nonfunctional (NF) pituitary adenomas, but the mechanistic role of FR{alpha} has not fully been determined. We investigated the effect of FR{alpha} over-expression in the mouse gonadotroph {alpha}T3-1 cell line as a model for NF pituitary adenomas. We found that the expression and function of FR{alpha} were strongly up-regulated, by Western blotting and folic acid binding assay. Furthermore, we found a higher cell growth rate, an enhanced percentage of cells in S-phase by BrdU assay, and a higher PCNA staining. These observations indicate that over-expression of FR{alpha} promotes cell proliferation. These effects were abrogated in the same {alpha}T3-1 cells when transfected with a mutant FR{alpha} cDNA that confers a dominant-negative phenotype by inhibiting folic acid binding. Finally, by real-time quantitative PCR, we found that mRNA expression of NOTCH3 was up-regulated in FR{alpha} over-expressing cells. In summary, our data suggests that FR{alpha} regulates pituitary tumor cell proliferation and mechanistically may involve the NOTCH pathway. Potentially, this finding could be exploited to develop new, innovative molecular targeted treatment for human NF pituitary adenomas.

  18. Analysis of the tool plunge in friction stir welding - comparison of aluminium alloys 2024 T3 and 2024 T351

    Directory of Open Access Journals (Sweden)

    Veljić Darko

    2016-01-01

    Full Text Available Temperature, plastic strain and heat generation during the plunge stage of the friction stir welding (FSW of high-strength aluminium alloys 2024 T3 and 2024 T351 are considered in this work. The plunging of the tool into the material is done at different rotating speeds. A three-dimensional finite element (FE model for thermomechanical simulation is developed. It is based on arbitrary Lagrangian-Eulerian formulation, and Johnson-Cook material law is used for modelling of material behaviour. From comparison of the numerical results for alloys 2024 T3 and 2024 T351, it can be seen that the former has more intensive heat generation from the plastic deformation, due to its higher strength. Friction heat generation is only slightly different for the two alloys. Therefore, temperatures in the working plate are higher in the alloy 2024 T3 for the same parameters of the plunge stage. Equivalent plastic strain is higher for 2024 T351 alloy, and the highest values are determined under the tool shoulder and around the tool pin. For the alloy 2024 T3, equivalent plastic strain is the highest in the influence zone of the tool pin. [Projekat Ministarstva nauke Republike Srbije, br. TR 34016 i br. TR 35006

  19. Piggy-back Hepatic Transplant Technique and Veno-venous Bypass Without Cardiac Arrest: A Multidisciplinary Approach in Borderline T3b/T3c Renal Tumors

    Directory of Open Access Journals (Sweden)

    Nechifor-Boila IA

    2015-06-01

    Full Text Available Surgery for renal cell carcinomas with tumor thrombus extending in the Inferior Vena Cava (IVC can be particularly challenging, especially in the retrohepatic and intraatrial situations (T3b and T3c. Classically, these tumors require the intraoperative use of cardio-pulmonary by-pass (CPB and deep hypothermic circulatory arrest (DHCA, that can result in specific complications (stroke, platelet dysfunction, with increased postoperative morbidity rates.

  20. Parallel magnetohydrodynamics on the Cray T3D

    NARCIS (Netherlands)

    Meijer, P. M.; Poedts, S.; Goedbloed, J. P.

    1996-01-01

    The equations of magnetohydrodynamics (MHD) are discussed in the framework of parallel computing. Both linear and nonlinear MHD models are addressed. Special attention is given to the parallellisation of the kernels of the existing sequential MHD codes. These kernels involve matrix-vector multiplica

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

    Energy Technology Data Exchange (ETDEWEB)

    Dong, Yan; Hirane, Miku; Araki, Mutsumi [Division of Cancer Biology and Bioinformatics, Department of Life Science, Faculty of Science and Engineering, Kinki University, 3-4-1, Kowakae, Higashiosaka, Osaka 577-8502 (Japan); Fukushima, Nobuyuki [Division of Molecular Neurobiology, Department of Life Science, Faculty of Science and Engineering, Kinki University, 3-4-1, Kowakae, Higashiosaka, Osaka 577-8502 (Japan); Tsujiuchi, Toshifumi, E-mail: ttujiuch@life.kindai.ac.jp [Division of Cancer Biology and Bioinformatics, Department of Life Science, Faculty of Science and Engineering, Kinki University, 3-4-1, Kowakae, Higashiosaka, Osaka 577-8502 (Japan)

    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}.

  2. Correlation between Serum T3 Resin Uptake and Serum Lipid Profile

    Directory of Open Access Journals (Sweden)

    Dinarvand, GH. ( MSc

    2015-05-01

    Full Text Available Background and Objective: T3 resin uptake test indicates the availability of thyroxin binding globulin (TBG, which is the carrier of a large amount of T3 and T4 in blood. We aimed to study the relationship between serum T3 Resin Uptake and serum lipids profile. Material and Methods: The participants aged 14.80 ± 38.29 were both males (59; 29.5% and females (141; 70.5 %. T3 Resin Uptake was measured by Immunoassay method and serum lipid profile components (cholesterol, triglycerides, HDL-C, LDL-C, and VLDL were measured by the kit. Results: Significant negative correlation was observed between serum T3 Resin Uptake and cholesterol, triglycerides and LDL-C (P≤ 0.05, but that was not the case for HDL-C and VLDL. Conclusion: Serum T3 Resin Uptake can be evaluated to assess the risk factors related to cardiovascular disorders

  3. Evaluation of chylomicron effect on ASP production in 3T3-L1 adipocytes

    Institute of Scientific and Technical Information of China (English)

    Ying Gao; Danny Gauvreau; Wei Cui; Marc Lapointe; Sabina Paglialunga; Katherine Cianflone

    2011-01-01

    In the past few years,there has been increasing interest in the production and physiological role of acylation-stimu-lating protein(ASP),identical to C3adesArg,a product of the alternative complement pathway generated through C3 cleavage.Recent studies in C3(-/-)mice that are ASP deficient have demonstrated a role for ASP in postprandial triglyceride clearance and fat storage.The aim of the present study was to establish a cell model and sensitive ELISA assay for the evaluation of ASP production using 3T3-L1 adipocytes.3T3-L1 preadipocytes were differentiated into adipocytes,then cultured in different media such as serum-free(SF),Dulbecco's modified Eagle's medium(DMEM)/F12+10% fetal calf serum (FBS),and at varying concentrations of chylomicrons and insulin+chylomicrons up to 48 h.ASP production in SF and DMEM/F12+10% FBS was compared.Chylomicrons stimulated ASP production in a concen-tration- and time-dependent manner.By contrast,chylo-micron treatment had no effect on the production of C3,the precursor protein of ASP,which was constant over 48 h.Addition of insulin(100 nM)to a low-dose of chylomicrons(100 μg TG/ml)significantly increased ASP production compared with chylomicrons alone at 48 h(P<0.001).Furthermore,addition of insulin significantly increased C3 secretion at both 18 and 48 h of incubation (P<0.05,P<0.001,respectively).Overall,the proportion of ASP to C3 remained constant,indicating no change in the ratio of C3 cleaved to generate ASP.This study demonstrated that 3T3-L1 adipocyte is a useful model for the evaluation of C3 secretion and ASP production by using a sensitive mouse-specific ELISA assay.The stimulation of ASP production with chylomicrons demonstrates a physiologically relevant response,and provides a strategy for further studies on ASP production and function.

  4. T3 affects expression of collagen I and collagen cross-linking in bone cell cultures

    OpenAIRE

    Varga, F.; Rumpler, M.; Zoehrer, R.; Turecek, C.; Spitzer, S.; Thaler, R; Paschalis, E.P.; Klaushofer, K.

    2010-01-01

    Thyroid hormones (T3, T4) have a broad range of effects on bone, however, its role in determining the quality of bone matrix is poorly understood. In-vitro, the immortalized mouse osteoblast-like cell line MC3T3-E1 forms a tissue like structure, consisting of several cell layers, whose formation is affected by T3 significantly. In this culture system, we investigated the effects of T3 on cell multiplication, collagen synthesis, expression of genes related to the collagen cross-linking process...

  5. Multiple Crack Growth Prediction in AA2024-T3 Friction Stir Welded Joints, Including Manufacturing Effects

    DEFF Research Database (Denmark)

    Carlone, Pierpaolo; Citarella, Roberto; Sonne, Mads Rostgaard

    2016-01-01

    boundary element method (FEM-DBEM) procedure, coupling the welding process simulation to the subsequent crack growth assessment, is proposed and applied to simulate multiple crack propagation, with allowance for manufacturing effects. The friction stir butt welding process of the precipitation hardened AA......2024-T3 alloy was simulated using a thermo-mechanical FEM model to predict the process induced residual stress field and material softening. The computed stress field was transferred to a DBEM environment and superimposed to the stress field produced by a remote fatigue traction load applied......A great deal of attention is currently paid by several industries toward the friction stir welding process to realize lightweight structures. Within this aim, the realistic prediction of fatigue behavior of welded assemblies is a key factor. In this work an integrated finite element method - dual...

  6. Multi-institutional analysis of T3 subtypes and adjuvant radiotherapy effects in resected T3N0 non-small cell lung cancer patients

    Energy Technology Data Exchange (ETDEWEB)

    Choi, Yun Seon; Lee, Ik Jae; Cho, Jae Ho [Dept. of Radiation Oncology, Severance Hospital, Yonsei University College of Medicine, Seoul (Korea, Republic of)

    2015-06-15

    We evaluated the prognostic significance of T3 subtypes and the role of adjuvant radiotherapy in patients with resected the American Joint Committee on Cancer stage IIB T3N0M0 non-small cell lung cancer (NSCLC). T3N0 NSCLC patients who underwent resection from January 1990 to October 2009 (n = 102) were enrolled and categorized into 6 subgroups according to the extent of invasion: parietal pleura chest wall invasion, mediastinal pleural invasion, diaphragm invasion, separated tumor nodules in the same lobe, endobronchial tumor <2 cm distal to the carina, and tumor-associated collapse. The median overall survival (OS) and disease-free survival (DFS) were 55.3 months and 51.2 months, respectively. In postoperative T3N0M0 patients, the tumor size was a significant prognostic factor for survival (OS, p = 0.035 and DFS, p = 0.035, respectively). Patients with endobronchial tumors within 2 cm of the carina also showed better OS and DFS than those in the other T3 subtypes (p = 0.018 and p = 0.016, respectively). However, adjuvant radiotherapy did not cause any improvement in survival (OS, p = 0.518 and DFS, p = 0.463, respectively). Only patients with mediastinal pleural invasion (n = 25) demonstrated improved OS and DFS after adjuvant radiotherapy (n = 18) (p = 0.012 and p = 0.040, respectively). The T3N0 NSCLC subtype that showed the most favorable prognosis is the one with endobronchial tumors within 2 cm of the carina. Adjuvant radiotherapy is not effective in improving survival outcome in resected T3N0 NSCLC.

  7. Triiodothyronine (T3)-associated upregulation and downregulation of nuclear T3 binding in the human fibroblast cell (MRC-5)--stimulation of malic enzyme, glucose-6-phosphate-dehydrogenase, and 6-phosphogluconate-dehydrogenase by insulin, but not by T3

    DEFF Research Database (Denmark)

    Matzen, L E; Kristensen, S R; Kvetny, J

    1991-01-01

    The specific nuclear binding of triiodothyronine (T3) (NBT3) and the activity of malic enzyme (ME), glucose-6-phosphate-dehydrogenase (G6PD), and 6-phosphogluconate-dehydrogenase (6PGD) were studied in the human fibroblast cell (MRC-5). The overall apparent binding affinity (Ka) was 2.7 x 10(9) L...

  8. Berberine reverses free-fatty-acid-induced insulin resistance in 3T3-L1 adipocytes through targeting IKKβ

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    AIM:To investigate the effects and molecular mechanisms of berberine on improving insulin resistance induced by free fatty acids (FFAs) in 3T3-LI adipocytes.METHODS:The model of insulin resistance in 3T3-L1 adipocytes was established by adding palmic acid (0.5 mmol/L) to the culture medium.Berberine treatment was performed at the same time.Glucose uptake rate was determined by the 2-deoxy-[3H]-Dglucose method.The levels of IkB kinase beta (IKKβ)Ser181 phosphorylation,insulin receptor substrate1(IRS-1) Ser307 phosphorylation,expression of IKKβ,IRS-1,nuclear transcription factor kappaB p65 (NF-κB p65),phosphatidylinositol-3-kinase p85(PI-3K p85) and glucose transporter 4 (GLUT4) proteins were detected by Western blotting.The distribution of NF-κB p65 proteins inside the adipocytes was observed through confocal laser scanning microscopy(CLSM).RESULTS:After the intervention of palmic acid for 24 h,the insulin-stimulated glucose transport in 3T3-L1 adipocytes was inhibited by 67%.Meanwhile,the expression of IRS-1 and PI-3K p85 protein was reduced,while the levels of IKKβ Ser181 and IRS-1 Ser307 phosphorylation,and nuclear translocation of NF-κB p65 protein were increased.However,the above indexes,which indicated the existence of insulin resistance,were reversed by berberine although the expression of GLUT4,IKKβ and total NF-κB p65 protein were not changed during this study.CONCLUSION:Insulin resistance induced by FFAs in 3T3-L1 adipocytes can be improved by berberine.Berberine reversed free-fatty-acid-induced insulin resistance in 3T3-L1 adipocytes through targeting IKKβ.

  9. The time development of hadronic showers and the T3B experiment

    Energy Technology Data Exchange (ETDEWEB)

    Soldner, Christian

    2013-06-06

    , the time of arrival of photons on the light sensor could be determined with sub-nanosecond precision. Embedded in a custom calibration and analysis framework, this allows for a precise study of shower timing on the nanosecond level. The T3B experiment could prove an increased contribution of the delayed shower component in tungsten with respect to steel via a detailed study of the time distribution of energy depositions. In addition, it is observed that the relative importance of late energy depositions increases with radial distance from the shower axis. This increase is substantially more pronounced in tungsten with respect to steel. It could be shown that the standard hadronic shower model QGSPBERT, used for shower simulations at the LHC as well as for most CLIC physics studies, overestimates the delayed shower evolution systematically, while high precision extensions using precise neutron tracking models can reproduce the shower timing adequately. No significant difference in the delayed shower contribution was observed for different particle energies in a range between 60 GeV and 180 GeV.

  10. T3i: A Tool for Generating and Querying Test Suites for Java

    NARCIS (Netherlands)

    Prasetya, I.S.W.B.|info:eu-repo/dai/nl/12691690X

    2015-01-01

    T3i is an automated unit-testing tool to test Java classes. To expose interactions T3i generates test-cases in the form of sequences of calls to the methods of the target class. What separates it from other testing tools is that it treats test suites as first class objects and allows users to e.g.

  11. T3i: A Tool for Generating and Querying Test Suites for Java

    NARCIS (Netherlands)

    Prasetya, I.S.W.B.

    2015-01-01

    T3i is an automated unit-testing tool to test Java classes. To expose interactions T3i generates test-cases in the form of sequences of calls to the methods of the target class. What separates it from other testing tools is that it treats test suites as first class objects and allows users to e.g. c

  12. T3, a Combinator-based Random Testing Tool for Java: Benchmarking

    NARCIS (Netherlands)

    Prasetya, I.S.W.B.

    2014-01-01

    T3 is the next generation of the light weight automated testing tool T2 for Java. In the heart T3 is still a random testing tool; but it now comes with some new features: pair-wise testing, concurrent generators, and a combinator-based approach ala QuickCheck. This paper presents the result of bench

  13. T24 HRAS transformed NIH/3T3 mouse cells (GhrasT-NIH/3T3) in serial tumorigenic in vitro/in vivo passages give rise to increasingly aggressive tumorigenic cell lines T1-A and T2-A and metastatic cell lines T3-HA and T4-PA.

    Science.gov (United States)

    Ray, Durwood B; Merrill, Gerald A; Brenner, Frederic J; Lytle, Laurie S; Lam, Tan; McElhinney, Aaron; Anders, Joel; Rock, Tara Tauber; Lyker, Jennifer Kier; Barcus, Scott; Leslie, Kara Hust; Kramer, Jill M; Rubenstein, Eric M; Pryor Schanz, Karen; Parkhurst, Amy J; Peck, Michelle; Good, Kimberly; Granath, Kristi Lemke; Cifra, Nicole; Detweiler, Jessalee Wantz; Stevens, Laura; Albertson, Richard; Deir, Rachael; Stewart, Elisabeth; Wingard, Katherine; Richardson, Micah Rose; Blizard, Sarah B; Gillespie, Lauren E; Kriley, Charles E; Rzewnicki, Daniel I; Jones, David H

    2016-01-01

    Cancer cells often arise progressively from "normal" to "pre-cancer" to "transformed" to "local metastasis" to "metastatic disease" to "aggressive metastatic disease". Recent whole genome sequencing (WGS) and spectral karyotyping (SKY) of cancer cells and tumorigenic models have shown this progression involves three major types of genome rearrangements: ordered small step-wise changes, more dramatic "punctuated evolution" (chromoplexy), and large catastrophic steps (chromothripsis) which all occur in random combinations to generate near infinite numbers of stochastically rearranged metastatic cancer cell genomes. This paper describes a series of mouse cell lines developed sequentially to mimic this type of progression. This starts with the new GhrasT-NIH/Swiss cell line that was produced from the NIH/3T3 cell line that had been transformed by transfection with HRAS oncogene DNA from the T24 human bladder carcinoma. These GhrasT-NIH/Swiss cells were injected s.c. into NIH/Swiss mice to produce primary tumors from which one was used to establish the T1-A cell line. T1-A cells injected i.v. into the tail vein of a NIH/Swiss mouse produced a local metastatic tumor near the base of the tail from which the T2-A cell line was established. T2-A cells injected i.v. into the tail vein of a nude NIH/Swiss mouse produced metastases in the liver and one lung from which the T3-HA (H=hepatic) and T3-PA (P=pulmonary) cell lines were developed, respectively. T3-HA cells injected i.v. into a nude mouse produced a metastasis in the lung from which the T4-PA cell line was established. PCR analysis indicated the human T24 HRAS oncogene was carried along with each in vitro/in vivo transfer step and found in the T2-A and T4-PA cell lines. Light photomicrographs indicate that all transformed cells are morphologically similar. GhrasT-NIH/Swiss cells injected s.c. produced tumors in 4% of NIH/Swiss mice in 6-10 weeks; T1-A cells injected s.c. produced tumors in 100% of NIH/Swiss mice in 7

  14. Low T3 syndrome is a strong prognostic predictor in diffuse large B cell lymphoma.

    Science.gov (United States)

    Gao, Rui; Liang, Jin-Hua; Wang, Li; Zhu, Hua-Yuan; Wu, Wei; Wu, Jia-Zhu; Xia, Yi; Cao, Lei; Fan, Lei; Yang, Tao; Li, Jian-Yong; Xu, Wei

    2017-02-01

    The aim of this study was to evaluate the prognostic effect of low triiodothyronine (T3) syndrome on patients with diffuse large B cell lymphoma (DLBCL). A hundred and eighty-eight patients with detailed thyroid hormone levels at diagnosis of DLBCL were enrolled. Low T3 syndrome was defined as a low serum free T3 (FT3) level with low or normal serum free tetraiodothyronine (FT4) and thyroid stimulating hormone levels. Multivariate Cox regression analysis was used to screen prognostic factors associated with progression-free survival (PFS) and overall survival (OS). Receiver-operator characteristic curves and the corresponding areas under the curve were calculated to assess the predictive accuracy of International Prognostic Index (IPI) and low T3 syndrome. Twenty-four patients were diagnosed with low T3 syndrome, which was associated with worse PFS and OS in the rituximab era. It was an independent prognostic factor for PFS and OS, especially for those with IPI 0-2, extranodal sites ≤1 and stage III-IV. Synchronously low FT3 and FT4 had poorer survival outcome compared to only low FT3 and adding criterion of low T3 syndrome improved the prognostic capacity of IPI for predicting PFS and OS in DLBCL. Low T3 syndrome was found to be a strong prognostic predictor in DLBCL.

  15. Sequence and analysis of the gene for bacteriophage T3 RNA polymerase.

    Science.gov (United States)

    McGraw, N J; Bailey, J N; Cleaves, G R; Dembinski, D R; Gocke, C R; Joliffe, L K; MacWright, R S; McAllister, W T

    1985-01-01

    The RNA polymerases encoded by bacteriophages T3 and T7 have similar structures, but exhibit nearly exclusive template specificities. We have determined the nucleotide sequence of the region of T3 DNA that encodes the T3 RNA polymerase (the gene 1.0 region), and have compared this sequence with the corresponding region of T7 DNA. The predicted amino acid sequence of the T3 RNA polymerase exhibits very few changes when compared to the T7 enzyme (82% of the residues are identical). Significant differences appear to cluster in three distinct regions in the amino-terminal half of the protein. Analysis of the data from both enzymes suggests features that may be important for polymerase function. In particular, a region that differs between the T3 and T7 enzymes exhibits significant homology to the bi-helical domain that is common to many sequence-specific DNA binding proteins. The region that flanks the structural gene contains a number of regulatory elements including: a promoter for the E. coli RNA polymerase, a potential processing site for RNase III and a promoter for the T3 polymerase. The promoter for the T3 RNA polymerase is located only 12 base pairs distal to the stop codon for the structural gene. PMID:3903658

  16. Aberrant CBFA2T3B gene promoter methylation in breast tumors

    Directory of Open Access Journals (Sweden)

    Bais Anthony J

    2004-08-01

    Full Text Available Abstract Background The CBFA2T3 locus located on the human chromosome region 16q24.3 is frequently deleted in breast tumors. CBFA2T3 gene expression levels are aberrant in breast tumor cell lines and the CBFA2T3B isoform is a potential tumor suppressor gene. In the absence of identified mutations to further support a role for this gene in tumorigenesis, we explored whether the CBFA2T3B promoter region is aberrantly methylated and whether this correlates with expression. Results Aberrant hypo and hypermethylation of the CBFA2T3B promoter was detected in breast tumor cell lines and primary breast tumor samples relative to methylation index interquartile ranges in normal breast counterpart and normal whole blood samples. A statistically significant inverse correlation between aberrant CBFA2T3B promoter methylation and gene expression was established. Conclusion CBFA2T3B is a potential breast tumor suppressor gene affected by aberrant promoter methylation and gene expression. The methylation levels were quantitated using a second-round real-time methylation-specific PCR assay. The detection of both hypo and hypermethylation is a technicality regarding the methylation methodology.

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

    Energy Technology Data Exchange (ETDEWEB)

    Lai, Peng-Yeh [Institute of Molecular Biology and Department of Life Science, National Chung Cheng University, Chiayi 621, Taiwan, ROC (China); Tsai, Chong-Bin [Institute of Molecular Biology and Department of Life Science, National Chung Cheng University, Chiayi 621, Taiwan, ROC (China); Department of Ophthalmology, Chiayi Christian Hospital, Chiayi 600, Taiwan, ROC (China); Tseng, Min-Jen, E-mail: biomjt@ccu.edu.tw [Institute of Molecular Biology and Department of Life Science, National Chung Cheng University, Chiayi 621, Taiwan, ROC (China)

    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.

  18. C2C12 myotubes inhibit the proliferation and differentiation of 3T3-L1 preadipocytes by reducing the expression of glucocorticoid receptor gene

    Energy Technology Data Exchange (ETDEWEB)

    Chu, Weiwei; Wei, Wei; Yu, Shigang; Han, Haiyin; Shi, Xiaoli [College of Animal Science and Technology, Nanjing Agricultural University, Nanjing 210095 (China); Sun, Wenxing [College of Animal Science and Technology, Nanjing Agricultural University, Nanjing 210095 (China); College of Public Health, Nantong University, Nantong 226019 (China); Gao, Ying [College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095 (China); Zhang, Lifan [College of Animal Science and Technology, Nanjing Agricultural University, Nanjing 210095 (China); Chen, Jie, E-mail: jiechen@njau.edu.cn [College of Animal Science and Technology, Nanjing Agricultural University, Nanjing 210095 (China)

    2016-03-25

    Obesity is a well-established risk factor to health for its relationship with insulin resistance, diabetes and metabolic syndrome. Myocyte-adipocyte crosstalk model plays a significant role in studying the interaction of muscle and adipose development. Previous related studies mainly focus on the effects of adipocytes on the myocytes activity, however, the influence of myotubes on the preadipocytes development remains unclear. The present study was carried out to settle this issue. Firstly, the co-culture experiment showed that the proliferation, cell cycle, and differentiation of 3T3-L1 preadipocytes were arrested, and the apoptosis was induced, by differentiated C2C12 myotubes. Next, the sensitivity of 3T3-L1 preadipocytes to glucocorticoids (GCs), which was well known as cell proliferation, differentiation, apoptosis factor, was decreased after co-cultured with C2C12 myotubes. What's more, our results showed that C2C12 myotubes suppressed the mRNA and protein expression of glucocorticoid receptor (GR) in 3T3-L1 preadipocytes, indicating the potential mechanism of GCs sensitivity reduction. Taken together, we conclude that C2C12 myotubes inhibited 3T3-L1 preadipocytes proliferation and differentiation by reducing the expression of GR. These data suggest that decreasing GR by administration of myokines may be a promising therapy for treating patients with obesity or diabetes. - Highlights: • C2C12 myotubes inhibited proliferation and differentiation of 3T3-L1 preadipocytes. • C2C12 myotubes arrested cell cycle of 3T3-L1 preadipocytes. • C2C12 myotubes induced apoptosis of 3T3-L1 preadipocytes. • C2C12 inhibit 3T3-L1 cells by reducing the expression of glucocorticoid receptor gene.

  19. The effects of Ganoderma lucidum herba pharmacopuncture on 3T3-L1 preadipocyte differentiation

    Directory of Open Access Journals (Sweden)

    Chea-woo Lee

    2008-09-01

    Full Text Available Objective : The purpose of this study is to investigate the effects of Ganoderma lucidum herba pharmacopuncture (GHP on the adipogenesis in 3T3-L1 preadipocytes. Methods : 3T3- L1 preadipocytes were differentiated with adipogenic reagents by incubating for 2 days in the absence or presence of GHP ranging from 1 and 2%. The effect of GHP on cell proliferation of 3T3-L1 preadipocytes was investigated using MTT assay. The effect of GHP on adipogenesis was examined by Oil red O staining and measuring glycerol-3-phosphate dehydrogenase (GPDH and intracellular triglyceride (TG content. Results : Following results were obtained from the preadipocyte proliferation and adipocyte differentiation of 3T3-L1. We observed no effect of GHP on preadipocyte proliferation. GHP inhibited adipogenesis, the activity of GPDH and accumulation of intracellular TG content. Conclusions : These results suggest that GHP inhibit differentiation of preadipocyte.

  20. Low T3 State Is Correlated with Cardiac Mitochondrial Impairments after Ischemia Reperfusion Injury: Evidence from a Proteomic Approach

    Directory of Open Access Journals (Sweden)

    Francesca Forini

    2015-11-01

    Full Text Available Mitochondria are major determinants of cell fate in ischemia/reperfusion injury (IR and common effectors of cardio-protective strategies in cardiac ischemic disease. Thyroid hormone homeostasis critically affects mitochondrial function and energy production. Since a low T3 state (LT3S is frequently observed in the post infarction setting, the study was aimed to investigate the relationship between 72 h post IR T3 levels and both the cardiac function and the mitochondrial proteome in a rat model of IR. The low T3 group exhibits the most compromised cardiac performance along with the worst mitochondrial activity. Accordingly, our results show a different remodeling of the mitochondrial proteome in the presence or absence of a LT3S, with alterations in groups of proteins that play a key role in energy metabolism, quality control and regulation of cell death pathways. Overall, our findings highlight a relationship between LT3S in the early post IR and poor cardiac and mitochondrial outcomes, and suggest a potential implication of thyroid hormone in the cardio-protection and tissue remodeling in ischemic disease.

  1. Hydroxytyrosol Inhibits Cannabinoid CB1 Receptor Gene Expression in 3T3-L1 Preadipocyte Cell Line.

    Science.gov (United States)

    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.

  2. Overexpression of GalNAc-transferase GalNAc-T3 promotes pancreatic cancer cell growth.

    Science.gov (United States)

    Taniuchi, K; Cerny, R L; Tanouchi, A; Kohno, K; Kotani, N; Honke, K; Saibara, T; Hollingsworth, M A

    2011-12-01

    O-linked glycans of secreted and membrane-bound proteins have an important role in the pathogenesis of pancreatic cancer by modulating immune responses, inflammation and tumorigenesis. A critical aspect of O-glycosylation, the position at which proteins are glycosylated with N-acetyl-galactosamine on serine and threonine residues, is regulated by the substrate specificity of UDP-GalNAc:polypeptide N-acetylgalactosaminyl-transferases (GalNAc-Ts). Thus, GalNAc-Ts regulate the first committed step in O-glycosylated protein biosynthesis, determine sites of O-glycosylation on proteins and are important for understanding normal and carcinoma-associated O-glycosylation. We have found that one of these enzymes, GalNAc-T3, is overexpressed in human pancreatic cancer tissues and suppression of GalNAc-T3 significantly attenuates the growth of pancreatic cancer cells in vitro and in vivo. In addition, suppression of GalNAc-T3 induces apoptosis of pancreatic cancer cells. Our results indicate that GalNAc-T3 is likely involved in pancreatic carcinogenesis. Modification of cellular glycosylation occurs in nearly all types of cancer as a result of alterations in the expression levels of glycosyltransferases. We report guanine the nucleotide-binding protein, α-transducing activity polypeptide-1 (GNAT1) as a possible substrate protein of GalNAc-T3. GalNAc-T3 is associated with O-glycosylation of GNAT1 and affects the subcellular distribution of GNAT1. Knocking down endogenous GNAT1 significantly suppresses the growth/survival of PDAC cells. Our results imply that GalNAc-T3 contributes to the function of O-glycosylated proteins and thereby affects the growth and survival of pancreatic cancer cells. Thus, substrate proteins of GalNAc-T3 should serve as important therapeutic targets for pancreatic cancers.

  3. Replication of Equine Infectious Anemia Virus in Engineered Mouse NIH 3T3 Cells ▿

    OpenAIRE

    Zhang, Baoshan; Montelaro, Ronald C.

    2008-01-01

    We employed the equine lentivirus equine infectious anemia virus (EIAV) to investigate the cellular restrictions for lentivirus replication in murine NIH 3T3 cells. The results of these studies demonstrate that NIH 3T3 cells expressing the EIAV receptor ELR1 and equine cyclin T1 supported productive replication of EIAV and produced infectious virions at levels similar to those found in a reference permissive equine cell line. The studies presented here demonstrate, for the first time, differe...

  4. High-dose Resveratrol Inhibits Insulin Signaling Pathway in 3T3-L1 Adipocytes

    OpenAIRE

    Lee, Haemi; Kim, Jae-Woo

    2013-01-01

    Background Insulin resistance is a major factor in the development of metabolic syndrome and is associated with central obesity and glucose intolerance. Resveratrol, a polyphenol found in fruits, has been shown to improve metabolic conditions. Although it has been widely studied how resveratrol affects metabolism, little is known about how resveratrol regulates lipogenesis with insulin signaling in 3T3-L1 adipocytes. Methods: We treated differentiated 3T3-L1 adipocytes with resveratrol to obs...

  5. T3 inhibits the calcification of vascular smooth muscle cells and the potential mechanism.

    Science.gov (United States)

    Chang, Xiaodan; Zhang, Baohong; Lihua, Li; Feng, Zhichun

    2016-01-01

    This study aimed to investigate the potential molecular mechanism underlying the T3 induced vascular calcification and phenotype transformation of vascular smooth muscle cells (VSMCs). Rat thoracic aortic smooth muscle cells (A7r5) were cultured in vitro and randomly assigned into normal control group, calcification group, T3 group and inhibitor group. When compared with normal control group, the osteocalcin content, ALP activity, Osterix and Runx2 mRNA expression and OPN protein expression increased significantly (P<0.01), and the protein expression of SMα and SM22α reduced dramatically in A7r5 cells of calcification group (P<0.01). After T3 treatment, the osteocalcin content and ALP activity reduced markedly, mRNA expression of Osterix and Runx2 and OPN protein expression reduced significantly. However, MMI (inhibitor of T3) was able to block the above effects of T3. When compared with calcification group, Osterix and Runx2 mRNA expression and OPN protein expression increased markedly (P<0.01). In addition, the protein expression of ERK1/2, p-ERK, Akt and p-Akt increased significantly in calcification group. In the presence of integrin αvβ3/ERK blocker (PD98059) and/or PI3K/Akt antagonist (LY294002), T3 was still able to inhibit the calcification, and this effect was similar to that after treatment with inhibitors alone. Moreover, LY294002 had a better inhibitory effect as compared to PD98059. T3 may act on PI3K/Akt signaling pathway to inhibit the phenotype transformation of VSMC, which then suppresses the calcium/phosphate induced calcification of rat VSMCs. Thus, T3 is an endogenous molecule that can protect the blood vessels against calcification.

  6. High-dose Resveratrol Inhibits Insulin Signaling Pathway in 3T3-L1 Adipocytes

    OpenAIRE

    Lee, Haemi; Kim, Jae-Woo

    2013-01-01

    Background Insulin resistance is a major factor in the development of metabolic syndrome and is associated with central obesity and glucose intolerance. Resveratrol, a polyphenol found in fruits, has been shown to improve metabolic conditions. Although it has been widely studied how resveratrol affects metabolism, little is known about how resveratrol regulates lipogenesis with insulin signaling in 3T3-L1 adipocytes. Methods: We treated differentiated 3T3-L1 adipocytes with resveratrol to obs...

  7. Uncoupling of 3T3-L1 gene expression from lipid accumulation during adipogenesis

    OpenAIRE

    Temple, Karla A.; Basko, Xheni; Allison, Margaret B.; Brady, Matthew J.

    2007-01-01

    Adipocyte differentiation comprises altered gene expression and increased triglyceride storage. To investigate the interdependency of these two events, 3T3-L1 cells were differentiated in the presence of glucose or pyruvate. All adipocytic proteins examined were similarly increased between the two conditions. In contrast, 3T3-L1 adipocytes differentiated with glucose exhibited significant lipid accumulation, which was largely suppressed in the presence of pyruvate. Subsequent addition of gluc...

  8. QUANTITATIVE STUDY ON APOPTOSIS INDUCED BY ELECTROMAGNETIC PULSES IN NIH- 3T3 FIBROBLASTS

    Institute of Scientific and Technical Information of China (English)

    ZHAO Mei - lan; CAO Xiao - zhe; WANG De - wen; GU Qing- yang; LIU Jie

    2002-01-01

    Aim: To observe the apoptotic changes following exposure to EMP and to explore the possible injury mechanism in NIH - 3T3 fibroblasts. Methods: Following NIH - 3T3 cells were exposed to EMP,the proliferation and viability of NIH - 3T3 fibroblasts were estimated by hemacytometer and MTT Measurements. Apoptotic rate was measured by flow cytometer. The imnmohistochemical SP method was used to determine bcl- 2, p53. The positively stained cells were analyzed with CMIAS- Ⅱ image analysis system at a magnification 400. All data were analyzed by SPSS8.0 software. Results: The proliferation and viability of NIH- 3T3 cells were markedly inhibited and significant apoptosis was induced following exposure to EMP.EMP could increase the number of non- adherent cells, the highest ratio (33.9%) of non- adherent cells also occurred at 6h. The As70 value of MTT decreased immediately at 1 h, 6h following the cells were exposed as compared with the control (P < 0.05). The highest ratio of apoptosis was 15.07% at 6h, then decreased gradually. Down - regulation of bcl - 2 and up - regulation of p53 were induced by EMP ( P < 0.05). Conclusions: EMP could promote apoptosis of NIH - 3T3 fibroblasts. EMP could also down - regulate bcl - 2 level and up - regulate p53 level in NIH - 3T3 fibroblasts. Bcl - 2 and p53 gene may take part in the process of apoptosis.

  9. Long-term physiological T3 supplementation in hypertensive heart disease in rats.

    Science.gov (United States)

    Weltman, Nathan Y; Pol, Christine J; Zhang, Youhua; Wang, Yibo; Koder, Adrienne; Raza, Sarah; Zucchi, Riccardo; Saba, Alessandro; Colligiani, Daria; Gerdes, A Martin

    2015-09-15

    Animal studies suggest that hypertension leads to cardiac tissue hypothyroidism, a condition that can by itself lead to heart failure. We have previously shown that short-term thyroid hormone treatment in Spontaneously Hypertensive Heart Failure (SHHF) rats near heart failure is beneficial. This study tested the hypothesis that therapeutic, long-term T3 treatment in SHHF rats can prevent or attenuate cardiac dysfunction. Female SHHF rats were treated orally with a physiological T3 dose (0.04 μg/ml) from 12 to 24 mo of age. Age-matched female SHHF and Wistar-Kyoto rats served as hypertensive and normotensive controls, respectively. SHHF rats had reduced serum free thyroid hormone levels and cardiac tissue T3 levels, LV dysfunction, and elevated LV collagen content compared with normotensive controls. Restoration of serum and cardiac tissue thyroid hormone levels in T3-treated rats was associated with no change in heart rate, but strong trends for improvement in LV systolic function and collagen levels. For instance, end-systolic diameter, fractional shortening, systolic wall stress, and LV collagen levels were no longer significantly different from controls. In conclusion, longstanding hypertension in rats led to chronic low serum and cardiac tissue thyroid hormone levels. Long-term treatment with low-dose T3 was safe. While cardiac dysfunction could not be completely prevented in the absence of antihypertensive treatment, T3 may offer additional benefits as an adjunct therapy with possible improvement in diastolic function. Copyright © 2015 the American Physiological Society.

  10. Commonalities and differences of T3SSs in rhizobia and plant pathogenic bacteria.

    Science.gov (United States)

    Tampakaki, Anastasia P

    2014-01-01

    Plant pathogenic bacteria and rhizobia infect higher plants albeit the interactions with their hosts are principally distinct and lead to completely different phenotypic outcomes, either pathogenic or mutualistic, respectively. Bacterial protein delivery to plant host plays an essential role in determining the phenotypic outcome of plant-bacteria interactions. The involvement of type III secretion systems (T3SSs) in mediating animal- and plant-pathogen interactions was discovered in the mid-80's and is now recognized as a multiprotein nanomachine dedicated to trans-kingdom movement of effector proteins. The discovery of T3SS in bacteria with symbiotic lifestyles broadened its role beyond virulence. In most T3SS-positive bacterial pathogens, virulence is largely dependent on functional T3SSs, while in rhizobia the system is dispensable for nodulation and can affect positively or negatively the mutualistic associations with their hosts. This review focuses on recent comparative genome analyses in plant pathogens and rhizobia that uncovered similarities and variations among T3SSs in their genetic organization, regulatory networks and type III secreted proteins and discusses the evolutionary adaptations of T3SSs and type III secreted proteins that might account for the distinguishable phenotypes and host range characteristics of plant pathogens and symbionts.

  11. Induction of adipocyte differentiation by polybrominated diphenyl ethers (PBDEs in 3T3-L1 cells.

    Directory of Open Access Journals (Sweden)

    Emily W Y Tung

    Full Text Available 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.

  12. Induction of adipocyte differentiation by polybrominated diphenyl ethers (PBDEs) in 3T3-L1 cells.

    Science.gov (United States)

    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.

  13. Anodic galvanostatic polarization of AA2024-T3 aircraft alloy in conventional mineral acids

    Energy Technology Data Exchange (ETDEWEB)

    Kozhukharov, S., E-mail: stephko1980@abv.bg [Department of Chemical Sciences, University of Chemical Technology and Metallurgy, 8 “Kliment Okhridski” Blvd, 1756, Sofia (Bulgaria); Girginov, Ch. [Department of Chemical Sciences, University of Chemical Technology and Metallurgy, 8 “Kliment Okhridski” Blvd, 1756, Sofia (Bulgaria); Avramova, I. [Institute of General and Inorganic Chemistry, Bulgarian Academy of Science, 11 “Georgi Bonchev” Str., 1113, Sofia (Bulgaria); Machkova, M. [Department of Chemical Sciences, University of Chemical Technology and Metallurgy, 8 “Kliment Okhridski” Blvd, 1756, Sofia (Bulgaria)

    2016-09-01

    The present study is devoted to the determination of the impact of the anodization of AA2024-T3 alloys in HCl, HNO{sub 3}, H{sub 2}SO{sub 4} or H{sub 3}PO{sub 4} on the samples’ surface morphology and properties. Subsequent systematic assessments were performed by Scanning Electron Microscopy (SEM), Energy Dispersion X-Ray Spectroscopy (EDX) and X-ray Photoelectron Spectroscopy (XPS). These observations were combined with Linear Voltammetry (LVA) and Electrochemical Impedance Spectroscopy (EIS) after 48 and 168 h of exposure to a 3.5% NaCl model corrosive medium. The main result is, that completely different effects were observed in accordance to the acid used. It was established that the monoprotonic acids have a deep destructive effect due to dissolution of the alloy components, whereas the polyprotonic ones possess either indistinguishable influence, or surface film formation. - Highlights: • AA2024 was polarized anodically in 15%{sub wt} acid solutions at 15 mA cm{sup −2} for 2 h. • Four mineral acids were selected for investigation: HCl, HNO{sub 3}, H{sub 2}SO{sub 4} and H{sub 3}PO{sub 4}. • SEM, EDX and XPS were applied for morphological description. • Electrochemical characterizations were performed by EIS and linear voltammetry. • The acid used predetermines completely different interaction with the AA2024 alloy.

  14. Effects of Methylmercury exposure in 3T3-L1 Adipocytes

    Directory of Open Access Journals (Sweden)

    Theresa Vertigan

    2017-02-01

    Full Text Available Mercury-containing compounds are environmental pollutants that have become increasingly consequential in the Arctic regions of North America due to processes of climate change increasing their release and availability at northern latitudes. Currently, the form of mercury known to be most detrimental to human health is methylmercury, CH3Hg+, which is found in the environment and accumulates in the tissues of piscivores, including those consumed by Alaska Natives through subsistence gathering. Much is known about the neurotoxicity of methylmercury after exposure to high concentrations, but little is known about toxicity to other tissues and cell types, particularly for long-term exposure and the lower concentrations that would occur through fish consumption. Effects of methylmercury exposure on 3T3-L1 adipocytes in culture were assessed using assays for cytotoxicity and an ELISA assay for vascular endothelial growth factor (VEGF, a signaling molecule shown to be important for maintaining metabolic status in adipose tissue. Results showed that exposure to methylmercury leads to significant toxicity in adipocytes at exposures of 100 ng/mL during later stages of differentiation, but lower methylmercury concentrations produced little to no toxicity. Results also showed that VEGF secretion is elevated in adipocytes exposed to methylmercury after the process of differentiating into mature, fat-storing cells. These results provide a basis for further exploration into metabolic consequences of methylmercury exposure on specific cell types and cell models.

  15. Illudins C2 and C3 stimulate lipolysis in 3T3-L1 adipocytes and suppress adipogenesis in 3T3-L1 preadipocytes.

    Science.gov (United States)

    Kim, Sun-Ok; Sakchaisri, Krisada; Asami, Yukihiro; Ryoo, In-Ja; Choo, Soo-Jin; Yoo, Ick-Dong; Soung, Nak-Kyun; Kim, Young Sang; Jang, Jae-Hyuk; Kim, Bo Yeon; Ahn, Jong Seog

    2014-04-25

    The secondary metabolites illudins C2 (1) and C3 (2), obtained from the culture broth of Coprinus atramentarius, have been shown to possess antimicrobial activity. In the present study, we discovered novel biological activities of 1 and 2 in lipolysis of differentiated 3T3-L1 adipocytes and adipogenesis of 3T3-L1 preadipocytes. Compounds 1 and 2 exhibit a dose-dependent increase in glycerol release and thereby reduce intracellular lipid accumulation. The stimulatory effects of 1 and 2 on lipolysis are prevented by cAMP-dependent protein kinase (PKA) and extracellular signal-regulated kinase (ERK) inhibitors. Compounds 1 and 2 down-regulated perilipin and also affected the mRNA and protein levels of adipose triglyceride lipase (ATGL) and hormone-sensitive lipase (HSL). However, 1 and 2 treatment leads to a significant increase in PKA-mediated phosphorylation of HSL at S563 and S660. In addition, 1 and 2 treatment in 3T3-L1 preadipocytes induces down-regulation of the critical transcription factors, CCAAT/enhancer binding protein α and β (C/EBPα and C/EBPβ), and peroxisome proliferator activated receptor γ (PPARγ), which are required for adipogenesis, and accordingly inhibits adipogenesis. These results suggest that 1 and 2 might be useful for treating obesity due to their modulatory effects on fat by affecting adipocyte differentiation and fat mobilization.

  16. Lipid droplets fusion in adipocyte differentiated 3T3-L1 cells: A Monte Carlo simulation

    Energy Technology Data Exchange (ETDEWEB)

    Boschi, Federico, E-mail: federico.boschi@univr.it [Department of Neurological and Movement Sciences, University of Verona, Strada Le Grazie 8, 37134 Verona (Italy); Department of Computer Science, University of Verona, Strada Le Grazie 15, 37134 Verona (Italy); Rizzatti, Vanni; Zamboni, Mauro [Department of Medicine, Geriatric Section, University of Verona, Piazzale Stefani 1, 37126 Verona (Italy); Sbarbati, Andrea [Department of Neurological and Movement Sciences, University of Verona, Strada Le Grazie 8, 37134 Verona (Italy)

    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

  17. Low T3 syndrome in canine babesiosis associated with increased serum IL-6 concentration and azotaemia.

    Science.gov (United States)

    Zygner, Wojciech; Gójska-Zygner, Olga; Bąska, Piotr; Długosz, Ewa

    2015-06-30

    Low triiodothyronine (T3) syndrome, also named euthyroid sick syndrome or non-thyroidal illness syndrome, has been recognized in canine babesiosis caused by Babesia rossi, where it manifested by lowering of the serum thyrotropin (TSH), total thyroxin (TT4) and free thyroxin (FT4) concentrations. This syndrome has also been observed in critical diseases in humans and animals, and the severity of the disease is considered an important factor in lowering of thyroid hormone concentrations. Interleukin-6 (IL-6) plays a role in the development of low T3 syndrome by causing a decrease in deiodinases 1 and 2 activity and increased activity of deiodinase 3, enzymes involved in the conversion of thyroxin (T4) to T3. The purpose of this study was to compare the concentrations of serum thyroid hormones and TSH between healthy dogs and dogs with babesiosis, and to determine correlations between serum IL-6 concentration and serum total T3 (TT3), TT4, FT4, and TSH concentrations, and the level of azotaemia in dogs with babesiosis. The concentrations of IL-6, TT3, TT4, FT4, TSH, urea and creatinine were determined in 13 dogs with canine babesiosis caused by Babesia canis and in 10 healthy dogs. The results of this study showed decreases in TT3, TT4, FT4, and TSH and increases in IL-6, urea and creatinine concentrations in affected dogs in comparison to healthy dogs. The concentration of IL-6 was negatively correlated with TT3 and TSH concentrations and the TT3 concentration was negatively correlated with serum urea and creatinine concentrations. This study showed low T3 syndrome in canine babesiosis, which was confirmed by the determination of the T3 concentration, and demonstrates that in canine babesiosis the T3 concentration is associated with IL-6 concentration. Copyright © 2015 Elsevier B.V. All rights reserved.

  18. Comparative studies on the structural proteins of T3 and T7 phages

    DEFF Research Database (Denmark)

    Issinger, O G; Falk, H

    1976-01-01

    A comparison of the coat protein patterns of the wild types of the related phages T3 and T7 on SDS polyacrylamide gel electrophoresis was carried out. After infection of the nonpermissive host with T7 amber mutants in genes 7, 11, 12, 13 or 17 and T3 amber mutants in genes 11, 12, 13 and 17...... respectively, noninfectious, DNA containing particles were produced. The protein pattern, as well as electron microscopy of defective particles of T3 and T7 led to the conclusion that the proteins specified by genes 11, 12, 13 and 17 are tail proteins whereas the proteins coded by genes 7, 8, 10, 14, 15 and 16...... enter the head structure. The "serum blocking protein" (gene 17 product) seems to play a different role in the assembly of T3 and T7 tails, since T3 particles defective in gene 17 did not show any tail structure connected with the head whereas T7 particles defective in gene 17 had a tail which looked...

  19. mdm-2 gene amplification in 3T3-L1 preadipocytes.

    Science.gov (United States)

    Berberich, S J; Litteral, V; Mayo, L D; Tabesh, D; Morris, D

    1999-05-01

    In this study the regulation of the murine double minute-2 (mdm-2) gene was examined in NIH 3T3-L1 preadipocytes. The 3T3-L1 cell line, under proper conditions, has the capacity to differentiate from fibroblasts into adipocytes [15]. A recent report demonstrated that mdm-2 overexpression could block myogenesis [12]. While examining the regulation of the mdm-2 gene during adipogenesis, it was discovered that 3T3-L1 cells possess a 36-fold elevation of mdm-2 mRNA relative to A31 cells, another immortalized Balb/c 3T3 fibroblast cell line that lacks the capacity to differentiate. Based on Southern blot analysis, the increase in mdm-2 mRNA was the result of a mdm-2 gene amplification. The level of Mdm-2 protein in undifferentiated 3T3-L1 cells was elevated relative to A31 fibroblasts and resulted from translation of mRNA transcripts initiating from the p53-independent P1 promoter. We also examined how mdm-2 and p53 levels changed as undifferentiated fibroblasts converted to adipocytes. While mdm-2 mRNA levels remained elevated, p53 mRNA, protein, and DNA-binding activity decreased. These results suggest that adipogenesis is unaffected by elevated Mdm-2 levels and that the overexpression of mdm-2 mRNA is predominantly p53 independent.

  20. Stair-climbing capabilities of USU's T3 ODV mobile robot

    Science.gov (United States)

    Robinson, D. Reed; Wood, Carl G.

    2001-09-01

    A six-wheeled autonomous omni-directional vehicle (ODV) called T3 has been developed at Utah State University's (USU) Center for Self-Organizing and Intelligent Systems (CSOIS). This paper focuses on T3's ability to climb stairs using its unique configuration of 6 independently driven and steered wheels and active suspension height control. The ability of T3, or any similar vehicle, to climb stairs is greatly dependent on the chassis orientation relative to the stairs. Stability criteria is developed for any vehicle dimensions and orientation, on any staircase. All possible yaw and pitch angles on various staircases are evaluated to find vehicle orientations that will allow T3 to climb with the largest margin of stability. Different controller types are investigated for controlling vertical wheel movement with the objective of keeping all wheels in contact with the stairs, providing smooth load transfer between loaded and unloaded wheels, and maintaining optimum chassis pitch and roll angles. A controller is presented that uses feedback from wheel loading, vertical wheel position, and chassis orientation sensors. The implementation of the controller is described, and T3's stair climbing performance is presented and evaluated.

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

    Energy Technology Data Exchange (ETDEWEB)

    Papadopoulos, T.; Pfeifer, U. (Univ. of Wuerzburg (West Germany))

    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.

  2. Cytotoxicity of Titanate-Calcium Complexes to MC3T3 Osteoblast-Like Cells

    Directory of Open Access Journals (Sweden)

    Yen-Wei Chen

    2016-01-01

    Full Text Available Monosodium titanates (MST are a relatively novel form of particulate titanium dioxide that have been proposed for biological use as metal sorbents or delivery agents, most recently calcium (II. In these roles, the toxicity of the titanate or its metal complex is crucial to its biological utility. The aim of this study was to determine the cytotoxicity of MST and MST-calcium complexes with MC3T3 osteoblast-like cells; MST-Ca(II complexes could be useful to promote bone formation in various hard tissue applications. MC3T3 cells were exposed to native MST or MST-Ca(II complexes for 24–72 h. A CellTiter-Blue® assay was employed to assess the metabolic activity of the cells. The results showed that MST and MST-Ca(II suppressed MC3T3 metabolic activity significantly in a dose-, time-, and cell-density-dependent fashion. MST-Ca(II suppressed MC3T3 metabolism in a statistically identical manner as native MST at all concentrations. We concluded that MST and MST-Ca(II are significantly cytotoxic to MC3T3 cells through a mechanism yet unknown; this is a potential problem to the biological utility of these complexes.

  3. Using Inertial Fusion Implosions to Measure the T + 3He Fusion Cross Section at Nucleosynthesis-Relevant Energies

    Science.gov (United States)

    Zylstra, A. B.; Herrmann, H. W.; Johnson, M. Gatu; Kim, Y. H.; Frenje, J. A.; Hale, G.; Li, C. K.; Rubery, M.; Paris, M.; Bacher, A.; Brune, C. R.; Forrest, C.; Glebov, V. Yu.; Janezic, R.; McNabb, D.; Nikroo, A.; Pino, J.; Sangster, T. C.; Séguin, F. H.; Seka, W.; Sio, H.; Stoeckl, C.; Petrasso, R. D.

    2016-07-01

    Light nuclei were created during big-bang nucleosynthesis (BBN). Standard BBN theory, using rates inferred from accelerator-beam data, cannot explain high levels of 6Li in in low-metallicity stars. Using high-energy-density plasmas we measure the T (3He, ,γ )6Li reaction rate, a candidate for anomalously high 6Li production; we find that the rate is too low to explain the observations, and different than values used in common BBN models. This is the first data directly relevant to BBN, and also the first use of laboratory plasmas, at comparable conditions to astrophysical systems, to address a problem in nuclear astrophysics.

  4. Using Inertial Fusion Implosions to Measure the T+^{3}He Fusion Cross Section at Nucleosynthesis-Relevant Energies.

    Science.gov (United States)

    Zylstra, A B; Herrmann, H W; Johnson, M Gatu; Kim, Y H; Frenje, J A; Hale, G; Li, C K; Rubery, M; Paris, M; Bacher, A; Brune, C R; Forrest, C; Glebov, V Yu; Janezic, R; McNabb, D; Nikroo, A; Pino, J; Sangster, T C; Séguin, F H; Seka, W; Sio, H; Stoeckl, C; Petrasso, R D

    2016-07-15

    Light nuclei were created during big-bang nucleosynthesis (BBN). Standard BBN theory, using rates inferred from accelerator-beam data, cannot explain high levels of ^{6}Li in low-metallicity stars. Using high-energy-density plasmas we measure the T(^{3}He,γ)^{6}Li reaction rate, a candidate for anomalously high ^{6}Li production; we find that the rate is too low to explain the observations, and different than values used in common BBN models. This is the first data directly relevant to BBN, and also the first use of laboratory plasmas, at comparable conditions to astrophysical systems, to address a problem in nuclear astrophysics.

  5. Calcification of MC3T3-E1 cells on titanium and zirconium.

    Science.gov (United States)

    Umezawa, Takayuki; Chen, Peng; Tsutsumi, Yusuke; Doi, Hisashi; Ashida, Maki; Suzuki, Shoichi; Moriyama, Keiji; Hanawa, Takao

    2015-01-01

    To confirm similarity of hard tissue compatibility between titanium and zirconium, calcification of MC3T3-E1 cells on titanium and zirconium was evaluated in this study. Mirror-polished titanium (Ti) and zirconium (Zr) disks and zirconium-sputter deposited titanium (Zr/Ti) were employed in this study. The surface of specimens were characterized using scanning electron microscopy and X-ray diffraction. Then, the cellular proliferation, differentiation and calcification of MC3T3-E1 cells on specimens were investigated. The surface of Zr/Ti was much smoother and cleaner than those of Ti and Zr. The proliferation of the cell was the same among three specimens, while the differentiation and calcification on Zr/Ti were faster than those on Ti and Zr. Therefore, Ti and Zr showed the identical hard tissue compatibility according to the evaluation with MC3T3-E1 cells. Sputter deposition may improve cytocompatibility.

  6. QUANTIFICATION OF SHEAR DAMAGE EVOLUTION IN ALUMINIUM ALLOY 2024T3

    Institute of Scientific and Technical Information of China (English)

    Tang Chak-yin; Fan Jianping; Tsui Chi-pong; Lee Tai-chiu; Chan Luen-chow; Rao Bin

    2007-01-01

    Shear damage may occur in the process of metal machining such as blanking and cutting, where localized shear deformation is developed. Experimental findings indicate that microscopic shear damage evolution in aluminium alloy 2024T3 (Al 2024T3) is a multi-stage mechanism, including particle cracking, micro-shear banding, matrix microcracking and coalescence of microcracks. This study is an attempt to use a set of equations to describe the multi-stage shear damage evolution in Al 2024T3. The shear damage variables in terms of multi-couple parameters of a power-law hardening material have been defined. An evolution curve of shearing damage has been calculated from experimental data. The values of the shear damage variable at different stages of damage have also been calculated. By making use of the findings, the relation between the microscopic shear damage evolution and the macroscopic shear response of the material has been discussed.

  7. NO-mediated regulation of GLUT by T3 and FSH in rat granulosa cells.

    Science.gov (United States)

    Tian, Ye; Ding, Yu; Liu, Juan; Heng, Dai; Xu, Kaili; Liu, Wenbo; Zhang, Cheng

    2017-03-17

    Thyroid hormones (THs) are important for normal reproductive function. Although 3,5,3'-triiodothyronine (T3) enhances follicle-stimulating hormone (FSH)-induced preantral follicle growth and granulosa cells development in vitro, little is known about the molecular mechanisms regulating ovarian development via glucose. In this study, we investigated whether and how T3 combines with FSH to regulate glucose transporter protein (GLUT) expression and glucose uptake in granulosa cells. Here, we present evidence that T3 and FSH co-treatment significantly increased GLUT-1/GLUT-4 expression, and translocation in cells, as well as glucose uptake. These changes were accompanied by upregulation of NOS3 expression, total NOS and NOS3 activity and NO content in granulosa cells. Furthermore, we found that activation of the mTOR and PI3K/Akt pathway is required for the regulation of GLUT expression, translocation, and glucose uptake by hormones. We also found that L-arginine (L-arg) up-regulated GLUT-1/GLUT-4 expression and translocation, which were related to increased glucose uptake, however, these responses were significantly blocked by L-NAME. In addition, inhibiting NO production attenuated T3 and FSH-induced GLUT expression, translocation, and glucose uptake in granulosa cells. Our data demonstrate that T3 and FSH co-treatment potentiates cellular glucose uptake via GLUT upregulation and translocation, which are mediated through the activation of the mTOR/PI3K/Akt pathway. Meanwhile, NOS3/NO are also involved in this regulatory system. These findings suggest that GLUT is a novel mediator of T3 and FSH-induced follicular development.

  8. Low T3 syndrome in neuromyelitis optica spectrum disorder: Associations with disease activity and disability.

    Science.gov (United States)

    Cho, Eun Bin; Min, Ju-Hong; Cho, Hye-Jin; Seok, Jin Myoung; Lee, Hye Lim; Shin, Hee Young; Lee, Kwang-Ho; Kim, Byoung Joon

    2016-11-15

    Neuromyelitis optica (NMO) sometimes coexists with serological marker-positive, non-organ-specific autoimmune disorders. We evaluated the prevalence of thyroid dysfunction and anti-thyroid antibodies in patients with NMO spectrum disorder (NMOSD) and investigated the associations between thyroid dysfunction/autoimmunity and clinical features of NMOSD. Forty-nine NMOSD patients with anti-aquaporin-4 antibody and 392 age- and sex-matched healthy controls were included. We measured the levels of thyroid hormones and anti-thyroid antibodies. The prevalence of clinical hypothyroidism, subclinical hyperthyroidism, and low T3 syndrome were higher in patients with NMOSD (4.1%, 12.2%, and 20.4%, respectively) compared with healthy controls (0.3%, 2.8%, and 0.5%, respectively; p=0.034, p=0.001, and p<0.001, respectively). However, anti-thyroperoxidase antibody (anti-TPO)-positivity did not significantly differ between NMOSD patients (20.4%) and controls (11.5%). Low T3 syndrome was more prevalent among patients during an attack (N=10/19, 52.6%) than those in remission (N=1/30, 3.3%). In addition, patients with low T 3 syndrome had significantly higher EDSS scores at the last visits as well as at sampling compared to those without low T3 syndrome. T3 levels were inversely correlated with EDSS score at the last visit after adjustment for age, sex, disease duration, clinical status (attack vs. remission), oral prednisolone use, iv methylprednisolone use, other immunosuppressive agents use, and the location of lesion (ρ=-0.416, p=0.010). Our study suggests that thyroid dysfunction is frequent in patients with NMOSD; particularly, serum T3 levels may be a useful indicator of disease activity and disability in NMOSD. Copyright © 2016 Elsevier B.V. All rights reserved.

  9. Establishment of an Enzyme Linked Immunosorbent Assay for Total Triiodothyronine (T3

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    A sensitive and specific ELISA for total triiodothyronine (T3) is established. The anh-T3 antibody iscoated on the microtiter plate, the T4 antigen is conjugated to the biotin. The label is horseradishperoxidase(HRP) conjugate of streptavidin. TMB-H2O2 solution is used as the substrate of HRP.Thesensitivity of the assay is 0.2 ng/mL, the intra-assay CVs and the intre-assay CVs of 3 samples are lower

  10. Uncoupling of 3T3-L1 gene expression from lipid accumulation during adipogenesis.

    Science.gov (United States)

    Temple, Karla A; Basko, Xheni; Allison, Margaret B; Brady, Matthew J

    2007-02-06

    Adipocyte differentiation comprises altered gene expression and increased triglyceride storage. To investigate the interdependency of these two events, 3T3-L1 cells were differentiated in the presence of glucose or pyruvate. All adipocytic proteins examined were similarly increased between the two conditions. In contrast, 3T3-L1 adipocytes differentiated with glucose exhibited significant lipid accumulation, which was largely suppressed in the presence of pyruvate. Subsequent addition of glucose to the latter cells restored lipid accumulation and acute rates of insulin-stimulated lipogenesis. These data indicate that extracellular energy is required for induction of adipocytic proteins, while only glucose sustained the parallel increase in triglyceride storage.

  11. File list: ALL.Oth.10.AllAg.3T3_fibroblasts [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  12. File list: His.Oth.50.AllAg.3T3_fibroblasts [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  13. File list: InP.Oth.20.AllAg.3T3_fibroblasts [Chip-atlas[Archive

    Lifescience Database Archive (English)

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    Lifescience Database Archive (English)

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  15. File list: His.EmF.10.AllAg.NIHSLASH3T3 [Chip-atlas[Archive

    Lifescience Database Archive (English)

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    Lifescience Database Archive (English)

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    Lifescience Database Archive (English)

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    Lifescience Database Archive (English)

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  1. File list: Oth.EmF.10.AllAg.NIHSLASH3T3 [Chip-atlas[Archive

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  9. File list: ALL.Oth.50.AllAg.3T3_fibroblasts [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  10. Mango fruit peel and flesh extracts affect adipogenesis in 3T3-L1 cells.

    Science.gov (United States)

    Taing, Meng-Wong; Pierson, Jean-Thomas; Hoang, Van L T; Shaw, Paul N; Dietzgen, Ralf G; Gidley, Michael J; Roberts-Thomson, Sarah J; Monteith, Gregory R

    2012-08-01

    Obesity is associated with many chronic disease states, such as diabetes mellitus, coronary disease and certain cancers, including those of the breast and colon. There is a growing body of evidence that links phytochemicals with the inhibition of adipogenesis and protection against obesity. Mangoes (Mangifera indica L.) are tropical fruits that are rich in a diverse array of bioactive phytochemicals. In this study, methanol extracts of peel and flesh from three archetypal mango cultivars; Irwin, Nam Doc Mai and Kensington Pride, were assessed for their effects on a 3T3-L1 pre-adipocyte cell line model of adipogenesis. High content imaging was used to assess: lipid droplets per cell, lipid droplet area per cell, lipid droplet integrated intensity, nuclei count and nuclear area per cell. Mango flesh extracts from the three cultivars did not inhibit adipogenesis; peel extracts from both Irwin and Nam Doc Mai, however, did so with the Nam Doc Mai extract most potent at inhibiting adipogenesis. Peel extract from Kensington Pride promoted adipogenesis. The inhibition of adipogenesis by Irwin (100 μg mL(-1)) and Nam Doc Mai peel extracts (50 and 100 μg mL(-1)) was associated with an increase in the average nuclear area per cell; similar effects were seen with resveratrol, suggesting that these extracts may act through pathways similar to resveratrol. These results suggest that differences in the phytochemical composition between mango cultivars may influence their effectiveness in inhibiting adipogenesis, and points to mango fruit peel as a potential source of nutraceuticals.

  11. 望京SOHO中心T3塔楼结构设计%Structural design on T3 tower of Wangjing SOHO Center

    Institute of Scientific and Technical Information of China (English)

    张克; 江坤生; 周坚荣; 朱勇军

    2013-01-01

    With the help of three-dimensional analysis software ETABS,three-dimensional nonlinear structural analysis software PERFORM-3D,and PKPM series software SATWE and PMSAP,and etc.,the structure of T3 tower of Wangjing SOHO Center was analyzed comprehensively.The structural dynamic response under the standard response spectrum and the actual seismic action and the plastic development situation of key parts and key components were analyzed.Seismic performance evaluation was carried out,and reasonable strengthening measures were proposed to the project design.%借助三维分析软件ETABS、三维非线性结构分析软件PERFORM-3D及PKPM系列软件SATWE和PMSAP等,对望京SOHO中心T3塔楼进行了全面的结构分析.分析了结构在规范反应谱及实际地震作用下的动力响应及关键部位、关键构件的塑性发展情况,对结构进行了抗震性能评价,并对工程设计提出合理的加强措施.

  12. Methylglyoxal induces oxidative stress and mitochondrial dysfunction in osteoblastic MC3T3-E1 cells.

    Science.gov (United States)

    Suh, K S; Choi, E M; Rhee, S Y; Kim, Y S

    2014-02-01

    Methylglyoxal is a reactive dicarbonyl compound produced by glycolytic processing and identified as a precursor of advanced glycation end products. The elevated methylglyoxal levels in patients with diabetes are believed to contribute to diabetic complications, including bone defects. The objective of this study was to evaluate the effect of methylglyoxal on the function of osteoblastic MC3T3-E1 cells. The data indicated that methylglyoxal decreased osteoblast differentiation and induced osteoblast cytotoxicity. Pretreatment of MC3T3-E1 cells with aminoguanidine (a carbonyl scavenger), Trolox (an antioxidant), and cyclosporin A (a blocker of the mitochondrial permeability transition pore) prevented methylglyoxal-induced cytotoxicity in MC3T3-E1 cells. However, BAPTA/AM (an intracellular Ca(2+) chelator) and dantrolene (an inhibitor of endoplasmic reticulum Ca(2+) release) did not reverse the cytotoxic effect of methylglyoxal. Methylglyoxal increased the formation of intracellular reactive oxygen species, mitochondrial superoxide, and cardiolipin peroxidation in osteoblastic MC3T3-E1 cells. Methylglyoxal also decreased the mitochondrial membrane potential and intracellular ATP and nitric oxide levels, suggesting that carbonyl stress-induced loss of mitochondrial integrity contributes to the cytotoxicity of methylglyoxal. Furthermore, the results demonstrated that methylglyoxal induced protein adduct formation, inactivation of glyoxalase I, and activation of glyoxalase II. Aminoguanidine reversed all aforementioned effects of methylglyoxal. Taken together, these data support the notion that high methylglyoxal concentrations have detrimental effects on osteoblasts through a mechanism involving oxidative stress and mitochondrial dysfunction.

  13. The Relationship between Selenium and T3 in Selenium Supplemented and Nonsupplemented Ewes and Their Lambs

    Directory of Open Access Journals (Sweden)

    Abd Elghany Hefnawy

    2014-01-01

    Full Text Available Twenty pregnant ewes were selected and classified into two groups. The first group received subcutaneous selenium supplementation (0.1 mg of sodium selenite/kg BW at the 8th and 5th weeks before birth and 1st week after birth while the other was control group without selenium injection. Maternal plasma and serum samples were collected weekly from the 8th week before birth until the 8th week after birth and milk samples were taken from ewes weekly, while plasma and serum samples were collected at 48 hours, 1st, 2nd, 3rd, 5th, and 8th weeks after birth from the newborn lambs. Results demonstrated significant positive relationship between maternal plasma selenium and serum T3 in supplemented and control ewes (r=0.69 to 0.72, P<0.05. There was significant (P<0.001 increase in T3 in supplemented ewes and their lambs until the 8th week after birth. There was positive relationship between milk, selenium concentration, and serum T3 in the newborn lambs of the supplemented group (r=0.84, P<0.01, while the relationship was negative in the control one (r=-0.89, P<0.01. Muscular and thyroid pathological changes were independent of selenium supplementation. Selenium supplementation was important for maintaining T3 in ewes and newborn lambs until the 8th week after birth.

  14. Increased Association of Dynamin Ⅱ with Myosin Ⅱ in Ras Transformed NIH3T3 Cells

    Institute of Scientific and Technical Information of China (English)

    Soon-Jeong JEONG; Su-Gwan KIM; Jiyun YOO; Mi-Young HAN; Joo-Cheol PARK; Heung-Joong KIM; Seong Soo KANG; Baik-Dong CHOI; Moon-Jin JEONG

    2006-01-01

    Dynamin has been implicated in the formation of nascent vesicles through both endocytic and secretory pathways. However, dynamin has recently been implicated in altering the cell membrane shape during cell migration associated with cytoskeleton-related proteins. Myosin Ⅱ has been implicated in maintaining cell morphology and in cellular movement. Therefore, reciprocal immunoprecipitation was carried out to identify the potential relationship between dynamin Ⅱ and myosin Ⅱ. The dynamin Ⅱ expression level was higher when co-expressed with myosin Ⅱ in Ras transformed NIH3T3 cells than in normal NIH3T3 cells.Confocal microscopy also confirmed the interaction between these two proteins. Interestingly, exposing the NIH3T3 cells to platelet-derived growth factor altered the interaction and localization of these two proteins.The platelet-derived growth factor treatment induced lamellipodia and cell migration, and dynamin Ⅱ interacted with myosin Ⅱ. Grb2, a 24 kDa adaptor protein and an essential element of the Ras signaling pathway,was found to be associated with dynamin Ⅱ and myosin Ⅱ gene expression in the Ras transformed NIH3T3 cells. These results suggest that dynamin Ⅱ acts as an intermediate messenger in the Ras signal transduction pathway leading to membrane ruffling and cell migration.

  15. Structure-activity relations in binding of perfluoroalkyl compounds to human thyroid hormone T3 receptor.

    Science.gov (United States)

    Ren, Xiao-Min; Zhang, Yin-Feng; Guo, Liang-Hong; Qin, Zhan-Fen; Lv, Qi-Yan; Zhang, Lian-Ying

    2015-02-01

    Perfluoroalkyl compounds (PFCs) have been shown to disrupt thyroid functions through thyroid hormone receptor (TR)-mediated pathways, but direct binding of PFCs with TR has not been demonstrated. We investigated the binding interactions of 16 structurally diverse PFCs with human TR, their activities on TR in cells, and the activity of perfluorooctane sulfonate (PFOS) in vivo. In fluorescence competitive binding assays, most of the 16 PFCs were found to bind to TR with relative binding potency in the range of 0.0003-0.05 compared with triiodothyronine (T3). A structure-binding relationship for PFCs was observed, where fluorinated alkyl chain length longer than ten, and an acid end group were optimal for TR binding. In thyroid hormone (TH)-responsive cell proliferation assays, PFOS, perfluorohexadecanoic acid, and perfluorooctadecanoic acid exhibited agonistic activity by promoting cell growth. Furthermore, similar to T3, PFOS exposure promoted expression of three TH upregulated genes and inhibited three TH downregulated genes in amphibians. Molecular docking analysis revealed that most of the tested PFCs efficiently fit into the T3-binding pocket in TR and formed a hydrogen bond with arginine 228 in a manner similar to T3. The combined in vitro, in vivo, and computational data strongly suggest that some PFCs disrupt the normal activity of TR pathways by directly binding to TR.

  16. 11 beta-hydroxysteroid dehydrogenase type 1 promotes differentiation of 3T3-L1 preadipocyte

    Institute of Scientific and Technical Information of China (English)

    Yun LIU; Yan SUN; Ting ZHU; Yu XIE; Jing YU; Wen-lan SUN; Guo-xian DING; Gang HU

    2007-01-01

    Aim: To investigate the relationship between 11 beta-hydroxysteroid dehydroge-nase type 1 (1 lbeta-HSD1), a potential link between obesity and type 2 diabetes,and preadipocyte differentiation. Methods: Mouse 11beta-HSD1 siRNA plasmids were transfected into 3T3-L1 preadipocytes (a cell line derived from mouse Swiss3T3 cells that were isolated from mouse embryo), for examination of the effect of targeted 11 beta-HSD1 inhibition on differentiation of 3T3-L1 cells. Dif-ferentiation was stimulated with 3-isobutyl-1-methyxanthine, insulin, and dexamethasone. The transcription level of the genes was detected by real-time PCR. Results: Lipid accumulation was significantly inhibited in cells transfected with mouse 11beta-HSD1 siRNA compared with non-transfected 3T3-L1 cells.Fewer lipid droplets were detected in the transfected cells both prior to stimulation and after stimulation with differentiation-inducing reagents. The expression of adipocyte differentiation-associated markers such as lipoprotein lipase and fatty acid synthetase were downregulated in the transfected cells. Similarly, the expres-sion of preadipocyte factor-1, an inhibitor of adipocyte differentiation, was downregulated upon stimulation of differentiation and had no changes in the transfected cells. Conclusion: 11 beta-HSD1 can promote preadipocyte differentiation. Based on this, we propose that 11 beta-HSD1 may be an important candidate mediator of obesity and obesity-induced insulin resistance.

  17. Preoperative chemoradiation of locally advanced T3 rectal cancer combined with an endorectal boost

    DEFF Research Database (Denmark)

    Jakobsen, Anders; Mortensen, John P; Bisgaard, Claus

    2006-01-01

    PURPOSE: To investigate the effect and feasibility of concurrent radiation and chemotherapy combined with endorectal brachytherapy in T3 rectal cancer with complete pathologic remission as end point. METHODS AND MATERIALS: The study included 50 patients with rectal adenocarcinoma. All patients had...

  18. Octanoate and decanoate induce apoptosis in 3T3-L1 adipocytes.

    Science.gov (United States)

    Yang, Jeong-Yeh; Rayalam, Srujana; Della-Fera, Mary Anne; Ambati, Suresh; Baile, Clifton A

    2009-10-01

    The effect of octanoate and decanoate, respectively, eight- and 10-carbon medium-chain fatty acids (MCFAs), on apoptotic signaling in 3T3-L1 adipocytes was investigated. 3T3-L1 adipocytes were treated with various concentrations of octanoate or decanoate. Cell viability, apoptosis, and expression of apoptosis-related proteins were investigated. Results indicated that both octanoate and decanoate decreased viability, increased apoptosis, and increased reactive oxygen species production. Immunoblotting analysis showed an increase in the levels of cytoplasmic cytochrome c and cleaved poly(ADP-ribose) polymerase by octanoate and decanoate. Concomitantly, we observed that pro-caspase-3 was decreased, resulting in the induced accumulation of the cleaved form of caspase-3 by both octanoate and decanoate. In addition, both octanoate and decanoate increased the expression of pro-apoptotic Bax with an accompanied decrease of anti-apoptotic Bcl-2. These results show that octanoate and decanoate mediate adipocyte apoptosis via a caspase-dependent mitochondrial pathway in 3T3-L1 adipocytes. MCFAs thus decrease adipocyte number by initiating the apoptotic process in 3T3-L1 adipocytes.

  19. Central diabetes insipidus preceding acute myeloid leukemia with t(3;12)(q26;p12)

    NARCIS (Netherlands)

    Nieboer, P; Vellenga, E; Adriaanse, R; van de Loosdrecht, AA

    2000-01-01

    A 52-year-old woman presented with polyuria and polydipsia. ii diagnosis of central diabetes insipidus (DI) was made, which turned out to be the first sign of acute myeloid leukemia (AML). Cytogenetic analysis revealed a balanced translocation between chromosome 3 and 12 t(3;12)(q26;p12). The patien

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

    Science.gov (United States)

    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.

  1. PREDICTION FOR FORMING LIMIT OF AL2024T3 SHEET BASED ON DAMAGE THEORY USING FINITE ELEMENT METHOD

    Institute of Scientific and Technical Information of China (English)

    Tang C.Y.; Fan Jianping; Tsui C.P.

    2006-01-01

    This paper presents the application of anisotropic damage theory to the study of forming limit diagram of Al2024T3 aluminum alloy sheet. In the prediction of limiting strains of the aluminum sheet structure, a finite element cell model has been constructed. The cell model consists of two phases, the aluminum alloy matrix and the intermetallic cluster. The material behavior of the aluminum alloy matrix is described with a fully coupled elasto-plastic damage constitutive equation. The intermetallic cluster is assumed to be elastic and brittle. By varying the stretching ratio, the limiting strains of the sheet under biaxial stretching have been predicted by using the necking criterion proposed. The prediction is in good agreement with the experimental findings. Moreover, the finite element cell model can provide information for understanding the microscopic damage mechanism of the aluminum alloy. Over-estimation of the limit strains may result if the effect of material damage is ignored in the sheet metal forming study.

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

    Science.gov (United States)

    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.

  3. Berberine activates GLUT1-mediated glucose uptake in 3T3-L1 adipocytes.

    Science.gov (United States)

    Kim, So Hui; Shin, Eun-Jung; Kim, Eun-Do; Bayaraa, Tsenguun; Frost, Susan Cooke; Hyun, Chang-Kee

    2007-11-01

    It has recently been known that berberine, an alkaloid of medicinal plants, has anti-hyperglycemic effects. To explore the mechanism underlying this effect, we used 3T3-L1 adipocytes for analyzing the signaling pathways that contribute to glucose transport. Treatment of berberine to 3T3-L1 adipocytes for 6 h enhanced basal glucose uptake both in normal and in insulin-resistant state, but the insulin-stimulated glucose uptake was not augmented significantly. Inhibition of phosphatidylinositol 3-kinase (PI 3-K) by wortmannin did not affect the berberine effect on basal glucose uptake. Berberine did not augment tyrosine phosphorylation of insulin receptor (IR) and insulin receptor substrate (IRS)-1. Further, berberine had no effect on the activity of the insulin-sensitive downstream kinase, atypical protein kinase C (PKCzeta/lambda). However, interestingly, extracellular signal-regulated kinases (ERKs), which have been known to be responsible for the expression of glucose transporter (GLUT)1, were significantly activated in berberine-treated 3T3-L1 cells. As expected, the level of GLUT1 protein was increased both in normal and insulin-resistant cells in response to berberine. But berberine affected the expression of GLUT4 neither in normal nor in insulin-resistant cells. In addition, berberine treatment increased AMP-activated protein kinase (AMPK) activity in 3T3-L1 cells, which has been reported to be associated with GLUT1-mediated glucose uptake. Together, we concluded that berberine increases glucose transport activity of 3T3-L1 adipocytes by enhancing GLUT1 expression and also stimulates the GLUT1-mediated glucose uptake by activating GLUT1, a result of AMPK stimulation.

  4. The effect of MAGE-A1 gene on NIH3T3 cells

    Institute of Scientific and Technical Information of China (English)

    Jingjun Sun; Jin Zhu; Zhenning Qiu; Yuhua Li; Guipeng Ding; Yi Zhu; Zhenqing Feng; Xiaohong Guan

    2005-01-01

    Objective: Melanoma antigen genes(MAGE) genes have been found in many kinds of tumor tissue, but not in normal tissue except testis and placentas. The Ags encoded by MAGE genes therefore are strictly tumor-specific. The most current researches associated with these genes focus on the tumor vaccination using these Ags. Few reports are concerning these genes' functions. In this study, we investigated the role of MAGE-A1 gene on NIH3T3 cells after transferring with it. Methods: Clone the MAGE-A1 into the plasmids pEGFP-C3 and pcDNA3.1, then transfer the reconstructed plasmids and primary plasmids into the NIH3T3 cells using a new transfer reagent FuGENE 6. Selecting the positively transferred cells by G418. Identified by RT-PCR, Western blot, Immunocytochemistry,Laser Scanning Confocal Microscope and Fluoroscope. The cells mobile ability was measured with Millicell-PCF. The cell cycle and apoptosis were measured with Flow Cytometry. Results: The apoptosis rate of NIH3T3 cells that transferred with control plasmid pcDNA3.1was 13.4% and the raitos that stay in S phase and G2-M phase were 5.68% and 1.04% respectively. The apoptosis rate of NIH3T3 cells that transferred with pcDNA3.1-A1 was 0.90% and the ratios that stayed in S phase and G2-M phase were 19.31% and 13.47% respectively. The apoptosis rate of the cells that transferred with control plasmid pEGFP-C3 was 1.87 %, a little higher than 1.47 % of those transferred with pEGFP-C3-A1. Conclusion: The MAGE-A1 gene may enhance the cell cycle, inhibit the apoptosis and raise the mobile ability of NIH3T3 cells.

  5. Environmental heat effects on growth, plasma T3, and postheat compensatory effects on Holstein calves.

    Science.gov (United States)

    Baccari, F; Johnson, H D; Hahn, G L

    1983-07-01

    Five Holstein heifers, 5 months of age, were housed in the Missouri Climatic Laboratory and subjected to an experiment to measure the effects of heat stress on rates of growth, plasma triiodothyronine (T3) levels, and ability to compensate in rate of gain and thyroid function following the stress period. The experiment consisted of 3 weeks at thermoneutral (TN1), followed by 5 weeks of individually controlled heat stress conditions (32.5 to 34 degrees C) dependent on heat tolerance of individual animals. This was followed by a 4-week thermoneutral, postheat compensatory period (TN2). Average daily gains were significantly depressed during the heat stress period (HS). Following heat stress the average body weights attained the projected or expected levels within a 21- to 28-day period following return of animals to thermoneutral conditions. Ratios of feed intake/body weight (w0.75) were reduced during heat stress treatment indicating the thermal inhibition. Ratios or amount of feed intake per unit of gain were greater during HS treatment indicating less weight gain per unit of daily feed intake. Plasma T3 was reduced during heat treatment similarly to daily weight gain. Following the postheat treatment period (TN2) plasma T3 increased markedly as did daily weight gains to demonstrate strong compensatory responses in both measures. In summary, these results demonstrated parallel and positive changes of plasma T3 with daily weight gain during thermoneutral, heat, and postheat compensatory periods, and an inverse relationship of rectal temperature to weight gain and plasma T3.

  6. Role of T3SS-1 SipD Protein in Protecting Mice against Non-typhoidal Salmonella Typhimurium

    Science.gov (United States)

    Jneid, Bakhos; Moreau, Karine; Plaisance, Marc; Rouaix, Audrey; Dano, Julie

    2016-01-01

    Background Salmonella enterica species are enteric pathogens that cause severe diseases ranging from self-limiting gastroenteritis to enteric fever and sepsis in humans. These infectious diseases are still the major cause of morbidity and mortality in low-income countries, especially in children younger than 5 years and immunocompromised adults. Vaccines targeting typhoidal diseases are already marketed, but none protect against non-typhoidal Salmonella. The existence of multiple non-typhoidal Salmonella serotypes as well as emerging antibiotic resistance highlight the need for development of a broad-spectrum protective vaccine. All Salmonella spp. utilize two type III Secretion Systems (T3SS 1 and 2) to initiate infection, allow replication in phagocytic cells and induce systemic disease. T3SS-1, which is essential to invade epithelial cells and cross the barrier, forms an extracellular needle and syringe necessary to inject effector proteins into the host cell. PrgI and SipD form, respectively, the T3SS-1 needle and the tip complex at the top of the needle. Because they are common and highly conserved in all virulent Salmonella spp., they might be ideal candidate antigens for a subunit-based, broad-spectrum vaccine. Principal Findings We investigated the immunogenicity and protective efficacy of PrgI and SipD administered by subcutaneous, intranasal and oral routes, alone or combined, in a mouse model of Salmonella intestinal challenge. Robust IgG (in all immunization routes) and IgA (in intranasal and oral immunization routes) antibody responses were induced against both proteins, particularly SipD. Mice orally immunized with SipD alone or SipD combined with PrgI were protected against lethal intestinal challenge with Salmonella Typhimurium (100 Lethal Dose 50%) depending on antigen, route and adjuvant. Conclusions and Significance Salmonella T3SS SipD is a promising antigen for the development of a protective Salmonella vaccine, and could be developed for

  7. Ginseng (Panax quinquefolius Reduces Cell Growth, Lipid Acquisition and Increases Adiponectin Expression in 3T3-L1 Cells

    Directory of Open Access Journals (Sweden)

    Chia-Rou Yeo

    2011-01-01

    Full Text Available An American ginseng (Panax quinquefolius extract (GE that contained a quantifiable amount of ginsenosides was investigated for the potential to inhibit proliferation, affect the cell cycle, influence lipid acquisition and adiponectin expression in 3T3-L1 cells. Six fingerprint ginsenosides were quantified by high performance liquid chromatography and the respective molecular weights were confirmed by LC-ESI-MS analysis. The extract contained Rg1 (347.3 ± 99.7 μg g−1, dry weight, Re (8280.4 ± 792.3 μg g−1, Rb1 (1585.8 ± 86.8 μg g−1, Rc (32.9 ± 8 μg g−1, Rb2 (62.6 ± 10.6 μg g−1 and Rd (90.4 ± 3.2 μg g−1. The GE had a dose-dependent effect on 3T3-L1 cell growth, the LC50 value was determined to be 40.3 ± 5 μg ml−1. Cell cycle analysis showed modest changes in the cell cycle. No significant changes observed in both G1 and G2/M phases, however there was a significant decrease (P<.05 in the S phase after 24 and 48 h treatment. Apoptotic cells were modest but significantly (P<.05 increased after 48 h (3.2 ± 1.0% compared to untreated control cells (1.5 ± 0.1%. Lipid acquisition was significantly reduced (P<.05 by 13 and 22% when treated at concentrations of 20.2 and 40.3 μg ml−1 compared to untreated control cells. In relation to adiponectin activation, western blot analysis showed that the protein expression was significantly (P<.05 increased at concentrations tested. A quantified GE reduced the growth of 3T3-L1 cells, down-regulated the accumulation of lipid and up-regulated the expression of adiponectin in the 3T3-L1 adipocyte cell model.

  8. Multiscale approach to micro/macro fatigue crack growth in 2024-T3 aluminum panel

    Science.gov (United States)

    Sih, G. C.

    2014-01-01

    When two contacting solid surfaces are tightly closed and invisible to the naked eye, the discontinuity is said to be microscopic regardless of whether its length is short or long. By this definition, it is not sufficient to distinguish the difference between a micro- and macro-crack by using the length parameter. Microcracks in high strength metal alloys have been known to be several centimeters or longer. Considered in this work is a dual scale fatigue crack growth model where the main crack can be micro or macro but there prevails an inherent microscopic tip region that is damaged depending on the irregularities of the microstructure. This region is referred to as the "micro-tip" and can be simulated by a sharp wedge with different angles in addition to mixed boundary conditions. The combination is sufficient to model microscopic entities in the form of voids, inclusions, precipitations, interfaces, in addition to subgrain imperfections, or cluster of dislocations. This is accomplished by using the method of "singularity representation" such that closed form asymptotic solutions can be obtained for the development of fatigue crack growth rate relations with three parameters. They include: (1) the crack surface tightness σ* represented by σ o/ σ ∞ = 0.3-0.5 for short cracks in region I, and 0.1-0.2 for long cracks in region II, (2) the micro/macro material properties reflected by the shear modulus ratio µ* (=µmicro/µmacro varying between 2 and 5) and (3) the most sensitive parameter d* being the micro-tip characteristic length d* (= d/ d o) whose magnitude decreases in the direction of region I→II. The existing fatigue crack growth data for 2024-T3 and 7075-T6 aluminum sheets are used to reinterpret the two-parameter d a/d N= C(Δ K) n relation where Δ K has now been re-derived for a microcrack with surfaces tightly in contact. The contact force will depend on the mean stress σm or mean stress ratio R as the primary parameter and on the stress

  9. Reassessment of the toxin profile of Cylindrospermopsis raciborskii T3 and function of putative sulfotransferases in synthesis of sulfated and sulfonated PSP toxins.

    Science.gov (United States)

    Soto-Liebe, Katia; Murillo, Alejandro A; Krock, Bernd; Stucken, Karina; Fuentes-Valdés, Juan J; Trefault, Nicole; Cembella, Allan; Vásquez, Mónica

    2010-12-01

    The toxigenic freshwater cyanobacterium Cylindrospermopsis raciborskii T3 has been used as a model to study and elucidate the biosynthetic pathway of tetrahydropurine neurotoxins associated with paralytic shellfish poisoning (PSP). There are nevertheless several inconsistencies and contradictions in the toxin profile of this strain as published by different research groups, and claimed to include carbamoyl (STX, NEO, GTX2/3), decarbamoyl (dcSTX), and N-sulfocarbamoyl (C1/2, B1) derivatives. Our analysis of the complete genome of another PSP toxin-producing cyanobacterium, Raphidiopsis brookii D9, which is closely related to C. raciborskii T3, resolved many issues regarding the correlation between biosynthetic pathways, corresponding genes and the T3 toxin profile. The putative sxt gene cluster in R. brookii D9 has a high synteny with the T3 sxt cluster, with 100% nucleotide identity among the shared genes. We also compared the PSP toxin profile of the strains by liquid chromatography coupled to mass spectrometry (LC-MS/MS). In contrast to published reports, our reassessment of the PSP toxin profile of T3 confirmed production of only STX, NEO and dcNEO. We gained significant insights via correlation between specific sxt genes and their role in PSP toxin synthesis in both D9 and T3 strains. In particular, analysis of sulfotransferase functions for SxtN (N-sulfotransferase) and SxtSUL (O-sulfotransferase) enzymes allowed us to propose an extension of the PSP toxin biosynthetic pathway from STX to the production of the derivatives GTX2/3, C1/2 and B1. This is a significantly revised view of the genetic mechanisms underlying synthesis of sulfated and sulfonated STX analogues in toxigenic cyanobacteria.

  10. The corrosion protection of AA2024-T3 aluminium alloy by leaching of lithium-containing salts from organic coatings.

    Science.gov (United States)

    Visser, Peter; Liu, Yanwen; Zhou, Xiaorong; Hashimoto, Teruo; Thompson, George E; Lyon, Stuart B; van der Ven, Leendert G J; Mol, Arjan J M C; Terryn, Herman A

    2015-01-01

    Lithium carbonate and lithium oxalate were incorporated as leachable corrosion inhibitors in model organic coatings for the protection of AA2024-T3. The coated samples were artificially damaged with a scribe. It was found that the lithium-salts are able to leach from the organic coating and form a protective layer in the scribe on AA2024-T3 under neutral salt spray conditions. The present paper shows the first observation and analysis of these corrosion protective layers, generated from lithium-salt loaded organic coatings. The scribed areas were examined by scanning and transmission electron microscopy before and after neutral salt spray exposure (ASTM-B117). The protective layers typically consist of three different layered regions, including a relatively dense layer near the alloy substrate, a porous middle layer and a flake-shaped outer layer, with lithium uniformly distributed throughout all three layers. Scanning electron microscopy and white light interferometry surface roughness measurements demonstrate that the formation of the layer occurs rapidly and, therefore provides an effective inhibition mechanism. Based on the observation of this work, a mechanism is proposed for the formation of these protective layers.

  11. The mixed-lineage kinase DLK is a key regulator of 3T3-L1 adipocyte differentiation.

    Directory of Open Access Journals (Sweden)

    Jean-Philippe Couture

    Full Text Available BACKGROUND: The mixed-lineage kinase (MLK family member DLK has been proposed to serve as a regulator of differentiation in various cell types; however, its role in adipogenesis has not been investigated. In this study, we used the 3T3-L1 preadipocyte cell line as a model to examine the function of DLK in adipocyte differentiation. METHODS AND FINDINGS: Immunoblot analyses and kinase assays performed on 3T3-L1 cells showed that the expression and activity of DLK substantially increase as differentiation occurs. Interestingly, DLK appears crucial for differentiation since its depletion by RNA interference impairs lipid accumulation as well as expression of the master regulators of adipogenesis C/EBPalpha and PPARgamma2 at both the mRNA and protein levels. In contrast, neither the expression nor the DNA binding activity of C/EBPbeta, an activator for C/EBPalpha and PPARgamma, is affected by DLK loss. CONCLUSIONS: Taken together, these results suggest that DLK is important for expression of mature adipocyte markers and that its action most likely takes place via regulation of C/EBPbeta transcriptional activity and/or initiation of C/EBPalpha and PPARgamma2 gene transcription.

  12. Transformation of BALB/c 3T3 cells in vitro by the fungicides captan, captafol and folpet.

    Science.gov (United States)

    Perocco, P; Colacci, A; Del Ciello, C; Grilli, S

    1995-10-01

    Cytotoxic and cell-transforming activities of the three fungicides, captan, captafol and folpet, have been studied in an experimental in vitro model by exposing BALB/c 3T3 cells to the chemicals with or without S-9 mix-induced bioactivation. Cytotoxicity of the three compounds was reduced in the presence of the metabolizing system. Each assayed pesticide displayed cell-transforming ability in the presence of the metabolizing system. The relative efficiency was: captafol > captan > folpet. Cell transformation was considered to be due to carcinogenesis-promoting activity. These data, obtained in a medium-term (6-8 weeks) experimental model, contribute to a better understanding of the action of the three pesticides in the multistep carcinogenesis process and provide more information concerning the oncogenic risk of these xenobiotic compounds for humans.

  13. Proof of the cosmic no-hair conjecture in the T^3-Gowdy symmetric Einstein-Vlasov setting

    CERN Document Server

    Andréasson, Håkan

    2013-01-01

    The currently preferred models of the universe undergo accelerated expansion induced by dark energy. One model for dark energy is a positive cosmological constant. It is consequently of interest to study Einstein's equations with a positive cosmological constant coupled to matter satisfying the ordinary energy conditions; the dominant energy condition etc. Due to the difficulty of analysing the behaviour of solutions to Einstein's equations in general, it is common to either study situations with symmetry, or to prove stability results. In the present paper, we do both. In fact, we analyse, in detail, the future asymptotic behaviour of T^3-Gowdy symmetric solutions to the Einstein-Vlasov equations with a positive cosmological constant. In particular, we prove the cosmic no-hair conjecture in this setting. However, we also prove that the solutions are future stable (in the class of all solutions). Some of the results hold in a more general setting. In fact, we obtain conclusions concerning the causal structure...

  14. SLC30A3 (ZnT3 oligomerization by dityrosine bonds regulates its subcellular localization and metal transport capacity.

    Directory of Open Access Journals (Sweden)

    Gloria Salazar

    Full Text Available Non-covalent and covalent homo-oligomerization of membrane proteins regulates their subcellular localization and function. Here, we described a novel oligomerization mechanism affecting solute carrier family 30 member 3/zinc transporter 3 (SLC30A3/ZnT3. Oligomerization was mediated by intermolecular covalent dityrosine bonds. Using mutagenized ZnT3 expressed in PC12 cells, we identified two critical tyrosine residues necessary for dityrosine-mediated ZnT3 oligomerization. ZnT3 carrying the Y372F mutation prevented ZnT3 oligomerization, decreased ZnT3 targeting to synaptic-like microvesicles (SLMVs, and decreased resistance to zinc toxicity. Strikingly, ZnT3 harboring the Y357F mutation behaved as a "gain-of-function" mutant as it displayed increased ZnT3 oligomerization, targeting to SLMVs, and increased resistance to zinc toxicity. Single and double tyrosine ZnT3 mutants indicate that the predominant dimeric species is formed between tyrosine 357 and 372. ZnT3 tyrosine dimerization was detected under normal conditions and it was enhanced by oxidative stress. Covalent species were also detected in other SLC30A zinc transporters localized in different subcellular compartments. These results indicate that covalent tyrosine dimerization of a SLC30A family member modulates its subcellular localization and zinc transport capacity. We propose that dityrosine-dependent membrane protein oligomerization may regulate the function of diverse membrane protein in normal and disease states.

  15. Gene expression analysis and microdialysis suggest hypothalamic triiodothyronine (T3) gates daily torpor in Djungarian hamsters (Phodopus sungorus).

    Science.gov (United States)

    Bank, Jonathan H H; Cubuk, Ceyda; Wilson, Dana; Rijntjes, Eddy; Kemmling, Julia; Markovsky, Hanna; Barrett, Perry; Herwig, Annika

    2017-07-01

    Thyroid hormones play an important role in regulating seasonal adaptations of mammals. Several studies suggested that reduced availability of 3,3',5-triiodothyronine (T3) in the hypothalamus is required for the physiological adaptation to winter in Djungarian hamsters. We have previously shown that T3 is involved in the regulation of daily torpor, but it remains unclear, whether T3 affects torpor by central or peripheral mechanisms. To determine the effect of T3 concentrations within the hypothalamus in regulating daily torpor, we tested the hypothesis that low hypothalamic T3 metabolism would favour torpor and high T3 concentrations would not. In experiment 1 gene expression in torpid hamsters was assessed for transporters carrying thyroid hormones between cerebrospinal fluid and hypothalamic cells and for deiodinases enzymes, activating or inactivating T3 within hypothalamic cells. Gene expression analysis suggests reduced T3 in hypothalamic cells during torpor. In experiment 2, hypothalamic T3 concentrations were altered via microdialysis and torpor behaviour was continuously monitored by implanted body temperature transmitters. Increased T3 concentrations in the hypothalamus reduced expression of torpor as well as torpor bout duration and depth. Subsequent analysis of gene expression in the ependymal layer of the third ventricle showed clear up-regulation of T3 inactivating deiodinase 3 but no changes in several other genes related to photoperiodic adaptations in hamsters. Finally, serum analysis revealed that increased total T3 serum concentrations were not necessary to inhibit torpor expression. Taken together, our results are consistent with the hypothesis that T3 availability within the hypothalamus significantly contributes to the regulation of daily torpor via a central pathway.

  16. Cell Volume Regulation and Signaling in 3T3-L1 Pre-adipocytes and Adipocytes

    DEFF Research Database (Denmark)

    Eduardsen, Kathrine; Larsen, Susanne; Novak, Ivana

    2011-01-01

    Caveolae have been implicated in sensing of cell volume perturbations, yet evidence is still limited and findings contradictory. Here, we investigated the possible role of caveolae in cell volume regulation and volume sensitive signaling in an adipocyte system with high (3T3-L1 adipocytes......) and osmotic cell shrinkage (RVI), accompanied by increased swelling-activated taurine efflux. However, caveolin-1 distribution was not detectably altered after osmotic swelling or shrinkage, and caveolae integrity, as studied by cholesterol depletion or expression of dominant negative Cav-1, was not required...... no detectable effect on InsR, yet increased ERK1/2 phosphorylation. In conclusion, differentiated 3T3-L1 adipocytes exhibit greatly accelerated RVD and RVI responses and increased swelling-activated taurine efflux compared to pre-adipocytes. Furthermore, in pre-adipocytes, Cav-1/caveolae integrity...

  17. Pleiotrophin Transforms NIH 3T3 Cells and Induces Tumors in Nude Mice

    Science.gov (United States)

    Chauhan, Anil K.; Li, Yue-Sheng; Deuel, Thomas F.

    1993-01-01

    The pleiotrophin (PTN) gene (Ptn) encodes an 18-kDa protein that is highly conserved among mammalian species and that functions as a weak mitogen and promotes neurite-outgrowth activity in vitro. To further investigate the role PTN plays in regulating cell growth, we overexpressed the bovine PTN cDNA and now show that PTN phenotypically transforms NIH 3T3 cells, as evidence by increased cell number at confluence, focus formation, anchorage-independent growth, and tumor formation in the nude muse. The results demonstrate that the Ptn gene has the potential to regulate NIH 3T3 cell growth and suggest that PTN may influence abnormal cell growth in vivo.

  18. Astrócitos e harmônio da tireóide (T3)

    OpenAIRE

    Aguiar, Cláudia Beatriz Nedel Mendes de

    2008-01-01

    Tese (doutorado) - Universidade Federal de Santa Catarina, Centro de Ciências Biológicas. Programa de Pós-graduação em Neurociências. O hormônio da tireóide (T3) exerce uma importante influência no desenvolvimento e maturação do SNC de mamíferos. Os efeitos do T3 no desenvolvimento neuronal são mediados pelos astrócitos, que secretam fatores solúveis e outras moléculas, como proteínas de MEC, modulando o crescimento de neuritos, proliferação e migração neuronal. Os astrócitos também partic...

  19. Evaluation of T3B fingerprinting for identification of clinical and environmental Sporothrix species.

    Science.gov (United States)

    Oliveira, Manoel Marques Evangelista; Franco-Duarte, Ricardo; Romeo, Orazio; Pais, Célia; Criseo, Giuseppe; Sampaio, Paula; Zancope-Oliveira, Rosely Maria

    2015-03-01

    In this study, PCR fingerprinting using the universal primer T3B was applied to distinguish among clinical and environmental species of the Sporothrix complex, Sporothrix brasiliensis, S. globosa, S. mexicana, S. pallida, S. luriei and S. schenckii sensu stricto. The T3B fingerprinting generated clearly distinct banding patterns, allowing the correct identification of all 43 clinical and environmental isolates at the species level, what was confirmed by partial calmodulin gene sequence analyses. This technique is reproducible and provides the identification of all species of the Sporothrix complex with sufficient accuracy to be applied in clinical mycology laboratories as well as in epidemiological studies in order to obtain a better understanding of the epidemiology of sporotrichosis.

  20. Revalidation of the isobaric multiplet mass equation at A=53, T=3/2

    Directory of Open Access Journals (Sweden)

    J. Su

    2016-05-01

    Full Text Available The T=3/2 isobaric analog state (IAS in 53Co is firmly established through a comprehensive measurement of β-delayed γ and proton decay of 53Ni. The determined excitation energy of 53Co IAS combined with the mass of 53Co generates a precise mass excess of −38333.6(27 keV for the 53Co IAS, which is 70(18 keV lower than the previously adopted value. The new result solves a problem raised by incorrect assignments of the 53Co IAS of unexpected deviation from the isobaric multiplet mass equation (IMME at A=53, T=3/2.

  1. Investigation of Droplet Deposition for Suspensions Usable for Thermoplastic 3D Printing (T3DP)

    Science.gov (United States)

    Scheithauer, Uwe; Johne, Robert; Weingarten, Steven; Schwarzer, Eric; Richter, Hans-Jürgen; Moritz, Tassilo; Michaelis, Alexander

    2017-09-01

    Thermoplastic 3D printing (T3DP) is an additive manufacturing (AM) technology, which can be used for the production of dense single- and especially multi-material components. This becomes possible because of the combination of the precise deposition of small droplets of molten thermoplastic suspensions containing ceramic or metal particles, and a curing mechanism caused on cool down increasing the viscosity. In this paper, the droplet formation behavior of zirconia suspensions for T3DP (82 and 84 wt.%) was investigated. The droplet fusion factor (dff) is introduced to calculate the necessary distance between two droplets to form filament-like structures by fusion of adjacent droplets. Filament-like structures with a smooth surface and a nearly homogeneous cross section were manufactured for both suspensions with a dff of 44% or higher.

  2. The intracellular mechanism of alpha-fetoprotein promoting the proliferation of NIH 3T3 cells

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    AIM The existence and properties of alpha-fetoprotein (AFP) receptor on the surface of NIH 3T3 cells and the effects of AFP on cellular signal transduction pathway were investigated. METHODS The effect of AFP on the proliferation of NIH 3T3 cells was measured by incorporation of 3H-TdR. Receptor-binding assay of 125I-AFP was performed to detect the properties of AFP receptor in NIH 3T3 cells. The influences of AFP on the [cAMP]i and the activities of protein kinase A (PKA) were determined. Western blot was used to detect the change of K-ras P21 protein expression. RESULTS The proliferation of NIH 3T3 cells treated with 0-80 mg/L of AFP was significantly enhanced. The Scatchard analysis indicated that there were two classes of binding sites with KD of 2.722×10-9M (Bmax=12810 sites per cell) and 8.931× 10-SM (Bmax=l19700 sites per cell) respectively. In the presence of AFP (20 mg/L), the content of cAMP and activities of PKA were significantly elevated . The level of K-ras P21 protein was upregulated by AFP at the concentration of 20 mg/L. The monoclonal antibody against AFP could reverse the effects of AFP on the cAMP content, PKA activity and the expression of K-ras p21 gene. CONCLUSION The effect of AFP on the cell proliferation was achieved by binding its receptor to trigger the signal transduction pathway of cAMP-PKA and alter the expression of K- ras p21 gene.

  3. Preoperative radiation therapy for upper rectal cancer T3,T4/Nx: selectivity essential.

    Science.gov (United States)

    Popek, Sarah; Tsikitis, Vassiliki Liana; Hazard, Lisa; Cohen, Alfred M

    2012-06-01

    This review explores the current available literature regarding the role of neoadjuvant therapy for upper locally advanced rectal cancers (≥10 cm-15 cm). Although there is a paucity of data evaluating the outcomes of preoperative chemoradiation for upper rectal cancers the authors suggest that T3N0 tumors will not likely benefit from radiation and that treatment of T4N0 should be individualized. Copyright © 2012. Published by Elsevier Inc.

  4. Comparative effect of Citrus sinensis and carbimazole on serum T 4 , T 3 and TSH levels

    Directory of Open Access Journals (Sweden)

    Okon Uduak Uduak

    2014-01-01

    Full Text Available Background: There are previous independent reports on the anti-thyroid property of Citrus sinensis. This isoflavones and phenolic acid-rich natural agent is widely consumed as dietary supplement, thus the need to investigate its comparative effect with a standard anti-thyroid drug on T 4 , T 3 and thyroid stimulating hormone (TSH levels. Objective: To compare the effect of Citrus sinensis and carbimazole (CARB on blood levels of thyroid hormones (T 4 and T 3 and TSH. Materials and Methods: Male wistar albino rats weighing 100-150 g were employed in this research. The rats were randomly assigned to four groups of seven rats per group. Group I served as control and were administered distilled water while groups II-IV were administered with 1500 mg/kg of Citrus sinensis (fresh orange juice; FOJ, 0.1 μg/g of levothyroxine (LVT and 0.01 mg/g of CARB, respectively, per oral once daily for 28 days. The animals were sacrificed under chloroform anaesthesia and blood sample collected by cardiac puncture and processed by standard method to obtain serum. TSH, T 4 and T 3 were assayed with the serum using ARIA II automated radioimmunoassay instrument. Results: The results showed that TSH level was significantly (P < 0.05 decreased in LVT treated group compared with the FOJ group. T 4 was significantly (P < 0.05 decreased in the FOJ and CARB groups compared with the control and LVT groups. LVT significantly increased T 4 when compared with FOJ group. T 3 was significantly (P < 0.05 decreased in the CARB group compared with the control. Conclusion: These findings suggest that FOJ alters thyroid hormones metabolism to reduce their serum levels with a compensatory elevations of TSH level in a direction similar to CARB.

  5. Stevioside from Stevia rebaudiana Bertoni Increases Insulin Sensitivity in 3T3-L1 Adipocytes

    Directory of Open Access Journals (Sweden)

    Nabilatul Hani Mohd-Radzman

    2013-01-01

    Full Text Available Stevioside from Stevia rebaudiana has been reported to exert antihyperglycemic effects in both rat and human subjects. There have been few studies on these effects in vitro. In this paper, radioactive glucose uptake assay was implemented in order to assess improvements in insulin sensitivity in 3T3-L1 cells by elevation of glucose uptake following treatment with stevioside. Oil Red-O staining and MTT assay were utilized to confirm adipocyte differentiation and cell viability, respectively. Findings from this research showed a significant increase in absorbance values in mature adipocytes following Oil Red-O staining, confirming the differentiation process. Stevioside was noncytotoxic to 3T3-L1 cells as cell viability was reduced by a maximum of 17%, making it impossible to determine its IC50. Stevioside increased glucose uptake activities by 2.1 times (p<0.001 in normal conditions and up to 4.4 times (p<0.001 in insulin-resistant states. At times, this increase was higher than that seen in positive control group treated with rosiglitazone maleate, an antidiabetic agent. Expressions of pY20 and p-IRS1 which were measured via Western blot were improved by stevioside treatment. In conclusion, stevioside has direct effects on 3T3-L1 insulin sensitivity via increase in glucose uptake and enhanced expression of proteins involved in insulin-signalling pathway.

  6. Rubi Fructus (Rubus coreanus) Inhibits Differentiation to Adipocytes in 3T3-L1 Cells.

    Science.gov (United States)

    Jeong, Mi-Young; Kim, Hye-Lin; Park, Jinbong; An, Hyo-Jin; Kim, Sung-Hoon; Kim, Su-Jin; So, Hong-Seob; Park, Raekil; Um, Jae-Young; Hong, Seung-Heon

    2013-01-01

    Rubi Fructus (RF) is known to exert several pharmacological effects including antitumor, antioxidant, and anti-inflammatory activities. However, its antiobesity effect has not been reported yet. This study was focused on the antidifferentiation effect of RF extract on 3T3-L1 preadipocytes. When 3T3-L1 preadipocytes were differentiating into adipocytes, 10-100  μ g/mL of RF was added. Next, the lipid contents were quantified by Oil Red O staining. RF significantly reduced lipid accumulation and downregulated the expression of peroxisome proliferator-activated receptor γ (PPAR γ ), CCAAT0-enhancer-binding proteins α (C/EBP α ), adipocyte fatty acid-binding protein 2 (aP2), resistin, and adiponectin in ways that were concentration dependent. Moreover, RF markedly upregulated liver kinase B1 and AMP-activated protein kinase (AMPK). Interestingly, pretreatment with AMPK α siRNA and RF downregulated the expression of PPAR γ and C/EBP α protein as well as the adipocyte differentiation. Our study shows that RF is capable of inhibiting the differentiation of 3T3-L1 adipocytes through the modulation of PPAR γ , C/EBP α , and AMPK, suggesting that it has a potential for therapeutic application in the treatment or prevention of obesity.

  7. T3/T4 thoracic sympathictomy and compensatory sweating in treatment of palmar hyperhidrosis

    Institute of Scientific and Technical Information of China (English)

    YANG Jie; TAN Jia-ju; YE Guo-lin; GU Wei-quan; WANG Jun; LIU Yan-guo

    2007-01-01

    Background Compensatory sweating (CS) is one of the most common postoperative complications after thoracic sympathectomy, sympathicotomy or endoscopic sympathetic block (ESB) for palmar hyperhidrosis. This study was conducted to examine the relevance between CS and the sympathetic segment being transected in the surgical treatment of palmar hyperhidrosis, and thus to detect the potential mechanism of the occurrence of CS.Methods Between October 2004 and June 2006, 163 patients with primary hyperhidrosis were randomly divided into two groups, T3 sympathicotomy (78 patients) and T4 sympathicotomy(85), who were operated upon under general anesthesia via single lumen intubation and intercostal video-mediastinoscopy (VM).Results No morbidity or mortality occurred. Palmar hyperhidrosis was cured in all patients. Follow-up(mean (13.8±6.2)months) showed no recurrence of palmar hyperhidrosis. The difference of rates of mild CS in groups T3 and T4 was of no statistical significance. The rate of moderate CS was significantly lower in group T4 than in group T3. No severe CS occurred.Conclusion The rates of occurrence and severity of CS are lowered with the lower sympathetic chain being transected.

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

    Science.gov (United States)

    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

  9. Effects of 6-Hydroxyflavone on Osteoblast Differentiation in MC3T3-E1 Cells

    Directory of Open Access Journals (Sweden)

    Chien-Hung Lai

    2014-01-01

    Full Text Available Osteoblast differentiation plays an essential role in bone integrity. Isoflavones and some flavonoids are reported to have osteogenic activity and potentially possess the ability to treat osteoporosis. However, limited information concerning the osteogenic characteristics of hydroxyflavones is available. This study investigates the effects of various hydroxyflavones on osteoblast differentiation in MC3T3-E1 cells. The results showed that 6-hydroxyflavone (6-OH-F and 7-hydroxyflavone (7-OH-F stimulated ALP activity. However, baicalein and luteolin inhibited ALP activity and flavone showed no effect. Up to 50 μM of each compound was used for cytotoxic effects study; flavone, 6-OH-F, and 7-OH-F had no cytotoxicity on MC3T3-E1 cells. Moreover, 6-OH-F activated AKT and serine/threonine kinases (also known as protein kinase B or PKB, extracellular signal-regulated kinases (ERK 1/2, and the c-Jun N-terminal kinase (JNK signaling pathways. On the other hand, 7-OH-F promoted osteoblast differentiation mainly by activating ERK 1/ 2 signaling pathways. Finally, after 5 weeks of 6-OH-F induction, MC3T3-E1 cells showed a significant increase in the calcein staining intensity relative to merely visible mineralization observed in cells cultured in the osteogenic medium only. These results suggested that 6-OH-F could activate AKT, ERK 1/2, and JNK signaling pathways to effectively promote osteoblastic differentiation.

  10. Cyclopia maculata (honeybush tea) stimulates lipolysis in 3T3-L1 adipocytes.

    Science.gov (United States)

    Pheiffer, Carmen; Dudhia, Zulfaqar; Louw, Johan; Muller, Christo; Joubert, Elizabeth

    2013-10-15

    We have previously, for the first time, demonstrated that hot water extracts of Cyclopia maculata and Cyclopia subternata, endemic South African plants that are consumed as herbal teas, inhibit adipogenesis in 3T3-L1 adipocytes. The aim of this study was to extend the anti-obesity investigations of these plants by quantifying lipolysis in mature 3T3-L1 adipocytes. Glycerol concentration in culture supernatants was used as a marker of adipocyte lipolysis. Isoproterenol, a β-adrenergic agonist and a known lipolytic agent, was used as a positive control in our assays. Lipolysis was stimulated by all extracts, although statistical significance was noted for fermented (oxidised) C. maculata only. A concentration of 80μg/ml of C. maculata extract induced maximal lipolysis (1.8-fold, plipolysis was accompanied by an increase in the expression of hormone sensitive lipase (1.6-fold, plipolysis in mature 3T3-L1 adipocytes, providing further support for the anti-obesity effects of Cyclopia spp. Copyright © 2013 Elsevier GmbH. All rights reserved.

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

    Energy Technology Data Exchange (ETDEWEB)

    Takahashi, Nobuhiko, E-mail: ntkhs@hoku-iryo-u.ac.jp [Department of Internal Medicine, School of Dentistry, Health Sciences University of Hokkaido, 1757, Kanazawa, Ishikari-Toubetsu, Hokkaido 061-0023 (Japan); Division of Gastroenterology and Hematology/Oncology, Department of Medicine, Asahikawa Medical University, 2-1-1-1, Midorigaoka-Higashi, Asahikawa, Hokkaido 078-8510 (Japan); Yoshizaki, Takayuki [Innovation Center, Kagoshima University, 1-21-40, Korimoto, Kagoshima 890-0065 (Japan); Hiranaka, Natsumi; Suzuki, Takeshi [Department of Internal Medicine, School of Dentistry, Health Sciences University of Hokkaido, 1757, Kanazawa, Ishikari-Toubetsu, Hokkaido 061-0023 (Japan); Yui, Tomoo; Akanuma, Masayoshi [Department of Fixed Prosthodontics and Oral Implantology, School of Dentistry, Health Sciences University of Hokkaido, 1757, Kanazawa, Ishikari-Toubetsu, Hokkaido 061-0023 (Japan); Kanazawa, Kaoru [Department of Dental Anesthesiology, School of Dentistry, Health Sciences University of Hokkaido, 1757, Kanazawa, Ishikari-Toubetsu, Hokkaido 061-0023 (Japan); Yoshida, Mika; Naito, Sumiyoshi [Department of Clinical Laboratory, Health Sciences University of Hokkaido, 1757, Kanazawa, Ishikari-Toubetsu, Hokkaido 061-0023 (Japan); Fujiya, Mikihiro; Kohgo, Yutaka [Division of Gastroenterology and Hematology/Oncology, Department of Medicine, Asahikawa Medical University, 2-1-1-1, Midorigaoka-Higashi, Asahikawa, Hokkaido 078-8510 (Japan); Ieko, Masahiro [Department of Internal Medicine, School of Dentistry, Health Sciences University of Hokkaido, 1757, Kanazawa, Ishikari-Toubetsu, Hokkaido 061-0023 (Japan)

    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.

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

    Directory of Open Access Journals (Sweden)

    Chian-Jiun Liou

    2015-01-01

    Full Text Available 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.

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

    Directory of Open Access Journals (Sweden)

    Sae-Rom Yoo

    2015-01-01

    Full Text Available 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.

  14. Evaluation of cytotoxicity and gelatinases activity in 3T3 fibroblast cell by root repair materials

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    Varol Basak

    2016-09-01

    Full Text Available The aim of this study was to investigate the effects of calcium silicate-based products on cytotoxicity in the 3T3 fibroblast and gelatinolytic activity of matrix metalloproteinases (MMPs. 3T3 fibroblasts were incubated directly with Ortho Mineral trioxide aggregate (MTA, BioAggregate, Biodentine, MTA Plus, MTA Angelus and MTA Cerkamed for 24 hours and seven days. The cytotoxicity was determined using an MTT assay. Supernatants were collected to determine MMP-2 and MMP-9. Data were analysed using IBM SPSS 22. Seventh day extracts of Ortho MTA and Biodentine showed reduced cell viability. Specific characterization of MMPs in cell culture demonstrated that MMP-2 (62 kPa in the cell culture supernatants by gelatin zymography showed induced expression in four out of seven groups by 3T3 cells. No MMP-9 expression was observed. The cytotoxicity of materials revealed a significant difference in cell viability between the groups on the first and seventh days. The results of this study revealed minor cytotoxic effects for Ortho MTA and Biodentine. This study suggests that endodontic sealers induced production of MMP-2. MMP-9 might be expressed in small amounts when compared with MMP-2.

  15. Aspirin Breaks the Crosstalk between 3T3-L1 Adipocytes and 4T1 Breast Cancer Cells by Regulating Cytokine Production.

    Directory of Open Access Journals (Sweden)

    Chia-Chien Hsieh

    Full Text Available Breast cancer is one of the most common cancers in women worldwide. The obesity process is normally accompanied by chronic, low-grade inflammation. Infiltration by inflammatory cytokines and immune cells provides a favorable microenvironment for tumor growth, migration, and metastasis. Epidemiological evidence has shown that aspirin is an effective agent against several types of cancer. The aim of this study is to investigate the anti-inflammatory and anti-cancer effects of aspirin on 3T3-L1 adipocytes, 4T1 murine breast cancer cells, and their crosstalk. The results showed that aspirin treatment inhibited differentiation and lipid accumulation by 3T3-L1 preadipocytes, and decreased the secretion of the inflammatory adipokine MCP-1 after stimulation with tumor necrosis factor (TNF-α or conditioned medium from RAW264.7 cells. In 4T1 cells, treatment with aspirin decreased cell viability and migration, possibly by suppressing MCP-1 and VEGF secretion. Subsequently, culture of 4T1 cells in 3T3-L1 adipocyte-conditioned medium (Ad-CM and co-culture of 3T3-L1 and 4T1 cells using a transwell plate were performed to clarify the relationship between these two cell lines. Aspirin exerted its inhibitory effects in the transwell co-culture system, as well as the conditioned-medium model. Aspirin treatment significantly inhibited the proliferation of 4T1 cells, and decreased the production of MCP-1 and PAI-1 in both the Ad-CM model and co-culture system. Aspirin inhibited inflammatory MCP-1 adipokine production by 3T3-L1 adipocytes and the cell growth and migration of 4T1 cells. It also broke the crosstalk between these two cell lines, possibly contributing to its chemopreventive properties in breast cancer. This is the first report that aspirin's chemopreventive activity supports the potential application in auxiliary therapy against obesity-related breast cancer development.

  16. Aspirin Breaks the Crosstalk between 3T3-L1 Adipocytes and 4T1 Breast Cancer Cells by Regulating Cytokine Production.

    Science.gov (United States)

    Hsieh, Chia-Chien; Huang, Yu-Shan

    2016-01-01

    Breast cancer is one of the most common cancers in women worldwide. The obesity process is normally accompanied by chronic, low-grade inflammation. Infiltration by inflammatory cytokines and immune cells provides a favorable microenvironment for tumor growth, migration, and metastasis. Epidemiological evidence has shown that aspirin is an effective agent against several types of cancer. The aim of this study is to investigate the anti-inflammatory and anti-cancer effects of aspirin on 3T3-L1 adipocytes, 4T1 murine breast cancer cells, and their crosstalk. The results showed that aspirin treatment inhibited differentiation and lipid accumulation by 3T3-L1 preadipocytes, and decreased the secretion of the inflammatory adipokine MCP-1 after stimulation with tumor necrosis factor (TNF)-α or conditioned medium from RAW264.7 cells. In 4T1 cells, treatment with aspirin decreased cell viability and migration, possibly by suppressing MCP-1 and VEGF secretion. Subsequently, culture of 4T1 cells in 3T3-L1 adipocyte-conditioned medium (Ad-CM) and co-culture of 3T3-L1 and 4T1 cells using a transwell plate were performed to clarify the relationship between these two cell lines. Aspirin exerted its inhibitory effects in the transwell co-culture system, as well as the conditioned-medium model. Aspirin treatment significantly inhibited the proliferation of 4T1 cells, and decreased the production of MCP-1 and PAI-1 in both the Ad-CM model and co-culture system. Aspirin inhibited inflammatory MCP-1 adipokine production by 3T3-L1 adipocytes and the cell growth and migration of 4T1 cells. It also broke the crosstalk between these two cell lines, possibly contributing to its chemopreventive properties in breast cancer. This is the first report that aspirin's chemopreventive activity supports the potential application in auxiliary therapy against obesity-related breast cancer development.

  17. Supraphysiological cyclic dosing of sustained release T3 in order to reset low basal body temperature.

    Science.gov (United States)

    Friedman, Michael; Miranda-Massari, Jorge R; Gonzalez, Michael J

    2006-03-01

    The use of sustained release tri-iodothyronine (SR-T3) in clinical practice, has gained popularity in the complementary and alternative medical community in the treatment of chronic fatigue with a protocol (WT3) pioneered by Dr. Denis Wilson. The WT3 protocol involves the use of SR-T3 taken orally by the patient every 12 hours according to a cyclic dose schedule determined by patient response. The patient is then weaned once a body temperature of 98.6 degrees F has been maintained for 3 consecutive weeks. The symptoms associated with this protocol have been given the name Wilson's Temperature Syndrome (WTS). There have been clinical studies using T3 in patients who are euthyroid based on normal TSH values. However, this treatment has created a controversy in the conventional medical community, especially with the American Thyroid Association, because it is not based on a measured deficiency of thyroid hormone. However, just as estrogen and progesterone are prescribed to regulate menstrual cycles in patients who have normal serum hormone levels, the WT3 therapy can be used to regulate metabolism despite normal serum thyroid hormone levels. SR-T3 prescription is based exclusively on low body temperature and presentation of symptoms. Decreased T3 function exerts widespread effects throughout the body. It can decrease serotonin and growth hormone levels and increase the number of adrenal hormone receptor sites. These effects may explain some of the symptoms observed in WTS. The dysregulation of neuroendocrine function may begin to explain such symptoms as alpha intrusion into slow wave sleep, decrease in blood flow to the brain, alterations in carbohydrate metabolism, fatigue, myalgia and arthralgia, depression and cognitive dysfunction. Despite all thermoregulatory control mechanisms of the body and the complex metabolic processes involved, WT3 therapy seems a valuable tool to re-establish normal body functions. We report the results of 11 patients who underwent the

  18. Increased thyroidal T4 to T3 conversion in autonomously functioning thyroid adenoma: from euthyroidism to thyrotoxicosis.

    LENUS (Irish Health Repository)

    Solter, M

    2012-01-31

    AIM: The aim was to investigate whether the intrathyroid conversion of T4 to T3 in autonomously functioning thyroid adenoma (AFTA) tissue could influence serum T3 levels and suppression of TSH, especially in patients with borderline thyroid function. PATIENTS AND METHODS: In ten patients with AFTA, thyroidal conversion of T4 to T3 was investigated in nodular and paranodular, TSH-suppressed tissue. All patients had normal serum T4 and suppressed TSH. Serum T3 was normal in six, and borderline or slightly increased in four. AFTA and paranodular tissues were surgically removed and frozen at -70 degrees C, then homogenized in a glass homogenizer, centrifuged at 100,000xg, and particulate fraction collected as a pellet. Analysis mixture consisted of thyroid enzyme suspension in 50 mumol\\/L TRIS buffer with 5 mumol DTT and 200 muL 1.3 mumol T4. Incubation was performed at 37 degrees C and the generation of T3 measured after 5, 10, 20 and 40 minutes respectively. RESULTS: T3 production (pmol\\/mg protein) was significantly higher in AFTA than in paranodular tissues (8.8 1.2\\/Mean +\\/- SE\\/vs. 1.8 +\\/- 0.2; p<0.01), and excessively high (9.8, 14.1, 14.2 and 15.0) in four patients with borderline or slightly supranormal serum T3. A significant correlation was found between serum T3 concentrations and T3 generation (T4 conversion) in AFTA tissues. CONCLUSION: Results suggest that increased thyroidal T4 to T3 conversion in AFTA tissue could be involved in an increased delivery of T3, increased serum T3 and suppressed serum TSH, particularly in patients with the disease evolving from euthyroid to an early hyperthyroid phase.

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    Lifescience Database Archive (English)

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    Lifescience Database Archive (English)

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  3. Comparison of bilateral T3 sympathectomy versus right T3 combined with left T4 sympa-thectomy in treatment of primary palmar hyperhidrosis%左 T4右 T3与双侧 T3交感神经链切断术治疗手汗症疗效比较

    Institute of Scientific and Technical Information of China (English)

    陈光春; 文毅; 廖小勇; 邱干; 郭伟华; 陈景松; 滕树炎; 陈洁; 吴维微

    2014-01-01

    Objective To evaluate the effect and side effect of right T 3 combined with left T 4 sympathectomy in the surgical management of primary palmar hyperhidrosis (PPH)through randomized controlled tria1.Methods From February 2012 to Apr 2014, 45 patients(19 males and 26 females)with PPH were enrolled in the study and randomly assigned into Group A (n=23) who recieved right T3 combined with left T4 sympathectomy and and Group B(n=22) who underwent bilateral T3 sympathectomy.The curative effects and side effects were compared between the tow groups .Results All procedures were carried out successfully without severe morbidity or mortality.After a mean follow-up period of 12.6 months,symptoms of PPH were improved in all patients .The left hands in Group A had more wet palms compared with the left hands in Group B (47.8%vs 9.1%,P0.05)and patients’ satisfaction (91.3%vs 95.5%,P>0.05)between Group A and B showed no statistically significant difference.Compensatory hyperhidrosis(CH) occurred in 66.7%of patients, and no patient was affected with severe CH .Moderate CH occurred in 3 cases and 9 cases respectively in groups A and B , and there was significant difference between the groups (13.0%vs 40.9%,P<0.05).Conclusions The procedure of right T3 combined with left T4 sympathectomy is an effective method which can improve quality of life and reduce compensatory hyperhidrosis .%目的:探讨左T4右T3交感神经链切断术治疗手汗症的可行性。方法2012-02至2014-04对45例手汗症行胸腔镜下胸交感神经链切断术,男19例,女26例,年龄15~42岁。患者自行选择哪种手术方式而随机进入两个组:A组为左T4右T3切断23例,B组为双侧T3链切断22例。结果所有手术均顺利完成,无严重并发症和围术期死亡病例。所有患者平均随访(12.6±9.5)个月。患者手掌多汗症状均改善。 A组9例(47.8%)的左侧手掌, A组2例(8.7%)和 B组2例(9.1%)的双侧手掌

  4. Thyroid of lake sturgeon, Acipenser fulvescens. II. Deiodination properties, distribution, and effects of diet, growth, and a T3 challenge.

    Science.gov (United States)

    Plohman, James C; Dick, Terry A; Eales, J Geoffrey

    2002-01-01

    The authors studied the properties and tissue distribution of thyroid hormone (TH) deiodination activities measured in vitro at subnanomolar substrate levels for cultured 2-year-old lake sturgeon held at 12 to 15 degrees. We also studied the deiodination responses to an exogenous 3,5,3'-triiodothyronine (T3) challenge and to a diet-induced growth suppression. Thyroxine (T4) outer-ring deiodination (T4ORD), T4 inner-ring deiodination (T4IRD), T3IRD, and 3,3',5'-triiodothyronine (rT3)ORD activities were evident in liver and intestine. Their properties resembled those of teleosts. T3IRD and T4IRD activities predominated in brain. Low or negligible deiodination in any form occurred in gill, skeletal muscle, kidney, notochord, or immature gonad. Only T4ORD activity was evident in the thyroid, suggesting that it secretes some T3. T3ORD and rT3IRD activities were undetectable in any tissues. Hepatic T4ORD activity varied during the photophase and was highest during late morning. A dietary T3 challenge that doubled plasma T3 levels decreased hepatic T4ORD activity without altering any other deiodination pathways in liver, intestine, or brain. A diet change from trout pellets to ocean zooplankton reduced somatic growth and plasma T3 levels and increased hepatic and intestinal T3IRD activities and hepatic rT3ORD activity but did not alter hepatic or intestinal T4ORD activity. The authors conclude that plasma T3 in lake sturgeon can be derived both from the thyroid and from hepatic (and intestinal) T4ORD activity, which varies with sampling time and downregulates in response to a T3 challenge. However, a reduction in plasma T3 accompanying a change in diet and reduced growth was not due to a decrease in T4ORD activity; rather, it was due to an increase in hepatic and intestinal T3IRD activities. These results suggest a difference in emphasis in thyroidal regulation between sturgeon and certain teleosts.

  5. Optical properties of hybrid T3Pyr/SiO2/3C-SiC nanowires.

    Science.gov (United States)

    Fabbri, Filippo; Rossi, Francesca; Melucci, Manuela; Manet, Ilse; Attolini, Giovanni; Favaretto, Laura; Zambianchi, Massimo; Salviati, Giancarlo

    2012-12-17

    A new class of nanostructured hybrid materials is developed by direct grafting of a model thiophene-based organic dye on the surface of 3C-SiC/SiO2 core/shell nanowires. TEM-EDX analysis reveals that the carbon distribution is more spread than it would be, considering only the SiC core size, suggesting a main contribution from C of the oligothiophene framework. Further, the sulfur signal found along the treated wires is not detected in the as-grown samples. In addition, the fluorescent spectra are similar for the functionalized nanostructures and T3Pyr in solution, confirming homogeneous molecule grafting on the nanowire surface. Chemical and luminescence characterizations confirm a homogeneous functionalization of the nanowires. In particular, the fluorophore retains its optical properties after functionalization.

  6. PPARγ partial agonist GQ-16 strongly represses a subset of genes in 3T3-L1 adipocytes

    Energy Technology Data Exchange (ETDEWEB)

    Milton, Flora Aparecida [Faculdade de Ciências da Saúde, Laboratório de Farmacologia Molecular, Universidade de Brasília (Brazil); Genomic Medicine, Houston Methodist Research Institute, Houston, TX (United States); Cvoro, Aleksandra [Genomic Medicine, Houston Methodist Research Institute, Houston, TX (United States); Amato, Angelica A. [Faculdade de Ciências da Saúde, Laboratório de Farmacologia Molecular, Universidade de Brasília (Brazil); Sieglaff, Douglas H.; Filgueira, Carly S.; Arumanayagam, Anithachristy Sigamani [Genomic Medicine, Houston Methodist Research Institute, Houston, TX (United States); Caro Alves de Lima, Maria do; Rocha Pitta, Ivan [Laboratório de Planejamento e Síntese de Fármacos – LPSF, Universidade Federal de Pernambuco (Brazil); Assis Rocha Neves, Francisco de [Faculdade de Ciências da Saúde, Laboratório de Farmacologia Molecular, Universidade de Brasília (Brazil); Webb, Paul, E-mail: pwebb@HoustonMethodist.org [Genomic Medicine, Houston Methodist Research Institute, Houston, TX (United States)

    2015-08-28

    Thiazolidinediones (TZDs) are peroxisome proliferator-activated receptor gamma (PPARγ) agonists that improve insulin resistance but trigger side effects such as weight gain, edema, congestive heart failure and bone loss. GQ-16 is a PPARγ partial agonist that improves glucose tolerance and insulin sensitivity in mouse models of obesity and diabetes without inducing weight gain or edema. It is not clear whether GQ-16 acts as a partial agonist at all PPARγ target genes, or whether it displays gene-selective actions. To determine how GQ-16 influences PPARγ activity on a gene by gene basis, we compared effects of rosiglitazone (Rosi) and GQ-16 in mature 3T3-L1 adipocytes using microarray and qRT-PCR. Rosi changed expression of 1156 genes in 3T3-L1, but GQ-16 only changed 89 genes. GQ-16 generally showed weak effects upon Rosi induced genes, consistent with partial agonist actions, but a subset of modestly Rosi induced and strongly repressed genes displayed disproportionately strong GQ-16 responses. PPARγ partial agonists MLR24 and SR1664 also exhibit disproportionately strong effects on transcriptional repression. We conclude that GQ-16 displays a continuum of weak partial agonist effects but efficiently represses some negatively regulated PPARγ responsive genes. Strong repressive effects could contribute to physiologic actions of GQ-16. - Highlights: • GQ-16 is an insulin sensitizing PPARγ ligand with reduced harmful side effects. • GQ-16 displays a continuum of weak partial agonist activities at PPARγ-induced genes. • GQ-16 exerts strong repressive effects at a subset of genes. • These inhibitor actions should be evaluated in models of adipose tissue inflammation.

  7. A resistin binding peptide selected by phage display inhibits 3T3-L1 preadipocyte differentiation

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    Background Resistin, a newly discovered cysteine-rich hormone secreted mainly by adipose tissues, has been proposed to form a biochemical link between obesity and type 2 diabetes. However, the resistin receptor has not yet been identified. This study aimed to identify resistin binding proteins/receptor.Methods Three cDNA fragments with the same 11 bp 5' sequence were found by screening a cDNA phage display library of rat multiple tissues. As the reading frames of the same 11 bp 5' sequence were interrupted by a TGA stop codon, plaque lift assay was consequently used to prove the readthrough phenomenon. The stop codon in the same 11 bp 5' sequence was replaced by tryptophan, and the binding activity of the coded peptide [AWIL, which was designated as resistin binding peptide (RBP)] with resistin was identified by the confocal microscopy technique and the affinity chromatography experiment. pDual GC-resistin and pDual GC-resistin binding peptide were co-transfected into 3T3-L1 cells to confirm the function of resistin binding peptide.Results Three cDNA fragments with the same 11 bp 5' sequence were found. The TGA stop codon in reading frames of the same 11 bp 5' sequence was proved to be readthroughed. The binding activity of RBP with resistin was consequently identified. The expression of the resistin binding peptide in 3T3-L1 preadipocytes expressing pDual GC-resistin significantly inhibited the adipogenic differentiation.Conclusion RBP could effectively rescue the promoted differentiation of resistin overxepressed 3T3-L1 preadipocyte.

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

    Energy Technology Data Exchange (ETDEWEB)

    Huang, Chenglin; Chen, Dongrui; Xie, Qihai [State Key Laboratory of Medical Genomics, Shanghai Key Laboratory of Vascular Biology, Department of Hypertension, Ruijin Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai 200025 (China); Yang, Ying, E-mail: yangying_sh@yahoo.com [Department of Endocrine and Metabolic Diseases, Shanghai Institute of Endocrine and Metabolic Diseases, Shanghai Clinical Center for Endocrine and Metabolic Diseases, Ruijin Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai 200025 (China); Shen, Weili, E-mail: weili_shen@hotmail.com [State Key Laboratory of Medical Genomics, Shanghai Key Laboratory of Vascular Biology, Department of Hypertension, Ruijin Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai 200025 (China)

    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.

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

    Science.gov (United States)

    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.

  10. Tea catechins modulate the glucose transport system in 3T3-L1 adipocytes.

    Science.gov (United States)

    Ueda, Manabu; Furuyashiki, Takashi; Yamada, Kayo; Aoki, Yukiko; Sakane, Iwao; Fukuda, Itsuko; Yoshida, Ken-Ichi; Ashida, Hitoshi

    2010-11-01

    In this study, we investigated the effects of tea catechins on the translocation of glucose transporter (GLUT) 4 in 3T3-L1 adipocytes. We found that the ethyl acetate fraction of green tea extract, containing abundant catechins, most decreased insulin-induced glucose uptake activity in 3T3-L1 cells. When the cells were treated with 50 μM catechins in the absence or presence of insulin for 30 min, nongallate-type catechins increased glucose uptake activity without insulin, whereas gallate-type catechins decreased insulin-induced glucose uptake activity. (-)-Epicatechin (EC) and (-)-epigallocatechin (EGC), nongallate-type catechins, increased glucose uptake activity in the dose- and time-dependent manner, whereas (-)-catechin 3-gallate (Cg) and (-)-epigallocatechin 3-gallate (EGCg), gallate-type catechins, decreased insulin-induced glucose uptake activity in the dose- and time-dependent manner. When the cells were treated with 50 μM catechins for 30 min, EC and EGC promoted GLUT4 translocation, whereas Cg and EGCg decreased the insulin-induced translocation in the cells. EC and EGC increased phosphorylation of PKCλ/ζ without phosphorylation of insulin receptor (IR) and Akt. Wortmannin and LY294002, inhibitors for phosphatidylinositol 3'-kinase (PI3K), decreased EC- and EGC-induced glucose uptake activity in the cells. Cg and EGCg decreased phosphorylation of PKCλ/ζ in the presence of insulin without affecting insulin-induced phosphorylation of IR, and Akt. Therefore, EC and EGC promote the translocation of GLUT4 through activation of PI3K, and Cg and EGCg inhibit insulin-induced translocation of GLUT4 by the insulin signaling pathway in 3T3-L1 cells.

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

    Energy Technology Data Exchange (ETDEWEB)

    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.

  12. Measurement of thyroid hormones (thyroxine, T4; triiodothyronine, T3) in captive nondomestic felids.

    Science.gov (United States)

    Rodini, Débora Cattaruzzi; Felippe, Erika Cristiane Gutierrez; Oliveira, Cláudio Alvarenga

    2007-03-01

    The aim of this research was to obtain basic values for the evaluation of thyroid function in nondomestic felids. Serum thyroid hormone concentrations (thyroxine, T4; triiodothyronine, T3) were measured by radioimmunoassay in 145 cats, representing nine species of captive nondomestic felids: jaguar (Panthera onca), n = 49; puma (Puma concolor), n = 10; ocelot (Leopardus pardalis), n = 22; oncilla (Leopardus tigrinus), n = 12; geoffroy (Oncifelis geoffroyi), n = 4; jaguarundi (Herpailurus yaguarondi), n = 8; margay (Leopardus wiedii), n = 7; lion (Panthera leo), n = 26; and tiger (Panthera tigris), n = 7. For each species, mean +/- SEM of T3 and T4, respectively, were as follows: jaguar, 0.56 +/- 0.03 and 9.7 +/- 0.8 ng/ml; puma, 0.67 +/- 0.04 and 11.2 +/- 1.2 ng/ml; ocelot, 0.48 +/- 0.03 and 13.8 +/- 1.5 ng/ml; oncilla, 0.43 +/- 0.03 and 10.0 +/- 1.6 ng/ml; geoffroy, 0.44 +/- 0.04 and 8.0 +/- 0.16 ng/ml; jaguarundi, 0.7 +/- 0.03 and 5.0 +/- 1.0 ng/ml; margay, 0.48 +/- 0.04 and 12.2 +/- 2.3 ng/ml; lion, 0.43 +/- 0.02 and 5.7 +/- 2.6 ng/ml; and tiger, 0.66 +/- 0.03 and 12.6 +/- 0.9 ng/ml. Within species, T3 and T4 concentrations did not differ (P > 0.05) between males and females.

  13. Effects of bovine colostral ultrafiltrates on growth and differentiation of 3T3-L1 preadipocytes.

    Science.gov (United States)

    Lee, Seong-Ho; Hossner, Kim L

    2002-12-01

    This study was designed to compare the effects of whole and size-fractionated bovine colostrum with bovine calf serum (BCS) on the growth and differentiation of 3T3-L1 fibroblasts. High (HMW) and low (LMW)-molecular-mass ultrafiltrate fractions of colostrum were prepared from defatted colostrum (COL) by diafiltration through membranes with a molecular-mass cut-off of 30 kDa. Incorporation of [(3)H]thymidine into the cells was used as a reflection of DNA synthesis/cell proliferation. The growth-promoting activity of LMW was 2.3- and 2.5-fold higher than COL and HMW, respectively (P insulin-like growth factor (IGF)-I, IGF-II and platelet-derived growth factor AB stimulated 3T3-L1 cells, antibodies to these factors did not inhibit the LMW effects. The LMW fraction was about twice as effective as COL and HMW in stimulating differentiation of the cells into adipocytes, but maximal differentiation was only 60% of that seen with 10% fetal bovine serum (FBS). Treatment with COL, HMW, IGF-I and insulin induced peroxisome-proliferator-activated receptor gamma RNA, but levels were about half of that with 10% FBS treatment and LMW induction was 80% of FBS. Low amounts of leptin mRNA were detected in adipocytes and abundance did not differ between treatments with BCS, hormones or COL fractions. This study showed that bovine colostral LMW stimulated the growth and differentiation of 3T3-L1 preadipocytes and may be a useful serum substitute to support the growth of these cells.

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

    Energy Technology Data Exchange (ETDEWEB)

    Cui, Ju, E-mail: juzi.cui@gmail.com [The Key Laboratory of Geriatrics, Beijing Hospital and Beijing Institute of Geriatrics, Beijing (China); Pang, Jing; Lin, Ya-Jun; Jiang, Ping; Gong, Huan [The Key Laboratory of Geriatrics, Beijing Hospital and Beijing Institute of Geriatrics, Beijing (China); Wang, Zai [Institute of Clinical Medical Sciences, China-Japan Friendship Hospital, Beijing (China); Li, Jian; Cai, Jian-Ping [The Key Laboratory of Geriatrics, Beijing Hospital and Beijing Institute of Geriatrics, Beijing (China); Huang, Jian-Dong, E-mail: jdhuang@hku.hk [School of Biomedical Sciences and Shenzhen Institute of Research and Innovation, The University of Hong Kong, Pokfulam (Hong Kong); The Centre for Synthetic Biology Engineering Research, Shenzhen Institutes of Advanced Technology, Shenzhen (China); Zhang, Tie-Mei, E-mail: tmzhang126@126.com [The Key Laboratory of Geriatrics, Beijing Hospital and Beijing Institute of Geriatrics, Beijing (China)

    2016-08-05

    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. - Highlights: • The KIF5B level was up regulated during 3T3-L1 adipogenesis. • Endogenous KIF5B and adiponectin were partially colicalized. • Adiponectin-containing vesicles were co-immunoprecipitated with KIF5B. • The secretion of adiponectin, but not leptin, is dependent on functional KIF5B.

  15. Hydrogen sulfide promotes adipogenesis in 3T3L1 cells.

    Directory of Open Access Journals (Sweden)

    Chin-Yi Tsai

    Full Text Available The effect of hydrogen sulfide (H2S on differentiation of 3T3L1-derived adipocytes was examined. Endogenous H2S was increased after 3T3L1 differentiation. The expression of the H2S-synthesising enzymes, cystathionine γ-lyase (CSE, cystathionine β-synthase (CBS and 3-mercaptopyruvate sulfurtransferase (3-MST, was increased in a time-dependent manner during 3T3L1 differentiation. Expression of genes associated with adipogenesis related genes including fatty acid binding protein 4 (FABP4/aP2, a key regulator of this process, was increased by GYY4137 (a slow-releasing H2S donor compound and sodium hydrosulfide (NaHS, a classical H2S donor but not by ZYJ1122 or time-expired NaHS. Furthermore expression of these genes were reduced by aminooxyacetic acid (AOAA, CBS inhibitor, DL-propargylglycine (PAG, CSE inhibitor as well as by CSE small interference RNA (siCSE and siCBS. The size and number of lipid droplets in mature adipocytes was significantly increased by both GYY4137 and NaHS, which also impaired the ability of CL316,243 (β3-agonist to promote lipolysis in these cells. In contrast, AOAA and PAG had the opposite effect. Taken together, we show that the H2S-synthesising enzymes CBS, CSE and 3-MST are endogenously expressed during adipogenesis and that both endogenous and exogenous H2S modulate adipogenesis and adipocyte maturation.

  16. Development of short fatigue cracks in aluminum alloy 2524-T3 specimens

    Science.gov (United States)

    Botvina, L. R.; Nesterenko, G. I.; Soldatenkov, A. P.; Demina, Yu. A.; Sviridov, A. A.

    2017-04-01

    The development of short fatigue cracks in a 2524-T3 alloy is studied under cyclic tension conditions. Flat specimens with a stress concentrator in the form of a central hole are analyzed. The replica technique is used to determine the microcrack parameters and to estimate the cyclic damage characteristics of the alloy in the stress concentrator zone. The experimental results are compared to the fatigue lives estimated by a calculation-experimental method using the NASGRO software package. The experimental fatigue life at the stage of short crack initiation is found to be significantly shorter than the calculated fatigue life.

  17. Nih-3T3 Fibroblast Studied by Fourier Transform Infrared Spectroscopy

    CERN Document Server

    Iovenitti, Marco

    2009-01-01

    In this work I present the study of the behaviour response of mouse fibroblasts NIH-3T3 under UVB radiation using Fourier transform infrared spectroscopy (FT-IR), the preferred method of infrared spectroscopy. FT-IR, in fact, it is convenient to study molecular cell processes because it has the potential to provide the identification of the vibrational modes of some of the major compounds (lipid, proteins and nucleic acids) without being invasive in the biomaterials. The results show that apoptotic process is induced by UVB radiation.

  18. Effects of dietary phytoestrogens on plasma testosterone and triiodothyronine (T3 levels in male goat kids

    Directory of Open Access Journals (Sweden)

    Ekstedt Elisabeth

    2009-12-01

    Full Text Available Abstract Background Exposure to xenoestrogens in humans and animals has gained increasing attention due to the effects of these compounds on reproduction. The present study was undertaken to investigate the influence of low-dose dietary phytoestrogen exposure, i.e. a mixture of genistein, daidzein, biochanin A and formononetin, on the establishment of testosterone production during puberty in male goat kids. Methods Goat kids at the age of 3 months received either a standard diet or a diet supplemented with phytoestrogens (3 - 4 mg/kg/day for ~3 months. Plasma testosterone and total and free triiodothyronine (T3 concentrations were determined weekly. Testicular levels of testosterone and cAMP were measured at the end of the experiment. Repeated measurement analysis of variance using the MIXED procedure on the generated averages, according to the Statistical Analysis System program package (Release 6.12, 1996, SAS Institute Inc., Cary, NC, USA was carried out. Results No significant difference in plasma testosterone concentration between the groups was detected during the first 7 weeks. However, at the age of 5 months (i.e. October 1, week 8 phytoestrogen-treated animals showed significantly higher testosterone concentrations than control animals (37.5 nmol/l vs 19.1 nmol/l. This elevation was preceded by a rise in plasma total T3 that occurred on September 17 (week 6. A slightly higher concentration of free T3 was detected in the phytoestrogen group at the same time point, but it was not until October 8 and 15 (week 9 and 10 that a significant difference was found between the groups. At the termination of the experiment, testicular cAMP levels were significantly lower in goats fed a phytoestrogen-supplemented diet. Phytoestrogen-fed animals also had lower plasma and testicular testosterone concentrations, but these differences were not statistically significant. Conclusion Our findings suggest that phytoestrogens can stimulate testosterone

  19. An MPI implementation of the SPAI preconditioner on the T3E

    Energy Technology Data Exchange (ETDEWEB)

    Barnard, Stephen T.; Bernardo, Luis M.; Simon, Horst D.

    1997-09-08

    The authors describe and test spai_1.1, a parallel MPIimplementation of the sparse approximate inverse (SPAI) preconditioner.They show that SPAI can be very effective for solving a set of very largeand difficult problems on a Cray T3E. The results clearly show the valueof SPAI (and approximate inverse methods in general) as the viablealternative to ILU-type methods when facing very large and difficultproblems. The authorsstrengthen this conclusion by showing that spai_1.1also has very good scaling behavior.

  20. T3PS v1.0: Tool for Parallel Processing in Parameter Scans

    Science.gov (United States)

    Maurer, Vinzenz

    2016-01-01

    T3PS is a program that can be used to quickly design and perform parameter scans while easily taking advantage of the multi-core architecture of current processors. It takes an easy to read and write parameter scan definition file format as input. Based on the parameter ranges and other options contained therein, it distributes the calculation of the parameter space over multiple processes and possibly computers. The derived data is saved in a plain text file format readable by most plotting software. The supported scanning strategies include: grid scan, random scan, Markov Chain Monte Carlo, numerical optimization. Several example parameter scans are shown and compared with results in the literature.

  1. Characterization of Catalase from Psychrotolerant Psychrobacter piscatorii T-3 Exhibiting High Catalase Activity

    OpenAIRE

    Hidetoshi Matsuyma; Isao Yumoto; Hideyuki Kimoto; Kazuaki Yoshimune

    2012-01-01

    A psychrotolerant bacterium, strain T-3 (identified as Psychrobacter piscatorii), that exhibited an extraordinarily high catalase activity was isolated from the drain pool of a plant that uses H2O2 as a bleaching agent. Its cell extract exhibited a catalase activity (19,700 U·mg protein−1) that was higher than that of Micrococcus luteus used for industrial catalase production. Catalase was approximately 10% of the total proteins in the cell extract of the strain. The catalase (PktA) was purif...

  2. Colchicine inhibits epidermal growth factor degradation in 3T3 cells.

    OpenAIRE

    Brown, K. D.; Friedkin, M; Rozengurt, E

    1980-01-01

    Colchicine (2 microM) did not affect the initial rate of association of 125I-labeled epidermal growth factor (125I-EGF) to Swiss 3T3 cells but continued incubation (up to 24 hr) led to an increase in cell-associated radioactivity. The effect is also produced by Colcemid, vinblastine, and podophyllotoxin but not by lumicolchicine. Disruption of microtubules with colchicine does not alter the rate of "down regulation" of EGF receptors, suggesting the binding and internalization of the factor pr...

  3. Overexpression of NYGGF4 (PID1) induces mitochondrial impairment in 3T3-L1 adipocytes.

    Science.gov (United States)

    Zhao, Yaping; Zhang, Chunmei; Chen, Xiaohui; Gao, Chunlin; Ji, Chenbo; Chen, Fukun; Zhu, Chun; Zhu, Jingai; Wang, Jialin; Qian, Lingmei; Guo, Xirong

    2010-07-01

    NYGGF4 is a recently discovered gene that is involved in obesity-associated insulin resistance. The exact mechanism by which NYGGF4 induces insulin resistance has not yet been fully elucidated. In this study, we demonstrated that the overexpression of NYGGF4 in 3T3-L1 adipocytes decreased mitochondrial mass, mitochondrial DNA, and intracellular ATP synthesis. In addition, NYGGF4 overexpression also led to an imbalance of the mitochondrial dynamics and excess intracellular ROS production. Collectively, our results indicated that the overexpression of NYGGF4 caused mitochondrial dysfunction in adipocytes, which might be responsible for the development of NYGGF4-induced insulin resistance.

  4. Berberine Alleviates Olanzapine-Induced Adipogenesis via the AMPKα-SREBP Pathway in 3T3-L1 Cells.

    Science.gov (United States)

    Li, Yanjie; Zhao, Xiaomin; Feng, Xiyu; Liu, Xuemei; Deng, Chao; Hu, Chang-Hua

    2016-11-09

    The aim of this study was to investigate the mechanisms underlying the inhibitory effects of berberine (BBR) on olanzapine (OLZ)-induced adipogenesis in a well-replicated 3T3-L1 cell model. Oil-Red-O (ORO) staining showed that BBR significantly decreased OLZ-induced adipogenesis. Co-treatment with OLZ and BBR decreased the accumulation of triglyceride (TG) and total cholesterol (TC) by 55.58% ± 3.65% and 49.84% ± 8.31%, respectively, in 3T3-L1 adipocytes accompanied by reduced expression of Sterol regulatory element binding proteins 1 (SREBP1), fatty acid synthase (FAS), peroxisome proliferator activated receptor-γ (PPARγ), SREBP2, low-density lipoprotein receptor (LDLR), and hydroxymethylglutaryl-coenzyme A reductase (HMGR) genes compared with OLZ alone. Consistently, the co-treatment downregulated protein levels of SREBP1, SREBP2, and LDLR by 57.71% ± 9.42%, 73.05% ± 11.82%, and 59.46% ± 9.91%, respectively. In addition, co-treatment reversed the phosphorylation level of AMP-activated protein kinase-α (AMPKα), which was reduced by OLZ, determined via the ratio of pAMPKα:AMPKα (94.1%) compared with OLZ alone. The results showed that BBR may prevent lipid metabolism disorders caused by OLZ by reversing the degree of SREBP pathway upregulated and the phosphorylation of AMPKα downregulated. Collectively, these results indicated that BBR could be used as a potential adjuvant to prevent dyslipidemia and obesity caused by the use of second-generation antipsychotic medication.

  5. Berberine Alleviates Olanzapine-Induced Adipogenesis via the AMPKα–SREBP Pathway in 3T3-L1 Cells

    Science.gov (United States)

    Li, Yanjie; Zhao, Xiaomin; Feng, Xiyu; Liu, Xuemei; Deng, Chao; Hu, Chang-Hua

    2016-01-01

    The aim of this study was to investigate the mechanisms underlying the inhibitory effects of berberine (BBR) on olanzapine (OLZ)-induced adipogenesis in a well-replicated 3T3-L1 cell model. Oil-Red-O (ORO) staining showed that BBR significantly decreased OLZ-induced adipogenesis. Co-treatment with OLZ and BBR decreased the accumulation of triglyceride (TG) and total cholesterol (TC) by 55.58% ± 3.65% and 49.84% ± 8.31%, respectively, in 3T3-L1 adipocytes accompanied by reduced expression of Sterol regulatory element binding proteins 1 (SREBP1), fatty acid synthase (FAS), peroxisome proliferator activated receptor-γ (PPARγ), SREBP2, low-density lipoprotein receptor (LDLR), and hydroxymethylglutaryl-coenzyme A reductase (HMGR) genes compared with OLZ alone. Consistently, the co-treatment downregulated protein levels of SREBP1, SREBP2, and LDLR by 57.71% ± 9.42%, 73.05% ± 11.82%, and 59.46% ± 9.91%, respectively. In addition, co-treatment reversed the phosphorylation level of AMP-activated protein kinase-α (AMPKα), which was reduced by OLZ, determined via the ratio of pAMPKα:AMPKα (94.1%) compared with OLZ alone. The results showed that BBR may prevent lipid metabolism disorders caused by OLZ by reversing the degree of SREBP pathway upregulated and the phosphorylation of AMPKα downregulated. Collectively, these results indicated that BBR could be used as a potential adjuvant to prevent dyslipidemia and obesity caused by the use of second-generation antipsychotic medication. PMID:27834848

  6. Berberine Alleviates Olanzapine-Induced Adipogenesis via the AMPKα–SREBP Pathway in 3T3-L1 Cells

    Directory of Open Access Journals (Sweden)

    Yanjie Li

    2016-11-01

    Full Text Available The aim of this study was to investigate the mechanisms underlying the inhibitory effects of berberine (BBR on olanzapine (OLZ-induced adipogenesis in a well-replicated 3T3-L1 cell model. Oil-Red-O (ORO staining showed that BBR significantly decreased OLZ-induced adipogenesis. Co-treatment with OLZ and BBR decreased the accumulation of triglyceride (TG and total cholesterol (TC by 55.58% ± 3.65% and 49.84% ± 8.31%, respectively, in 3T3-L1 adipocytes accompanied by reduced expression of Sterol regulatory element binding proteins 1 (SREBP1, fatty acid synthase (FAS, peroxisome proliferator activated receptor-γ (PPARγ, SREBP2, low-density lipoprotein receptor (LDLR, and hydroxymethylglutaryl-coenzyme A reductase (HMGR genes compared with OLZ alone. Consistently, the co-treatment downregulated protein levels of SREBP1, SREBP2, and LDLR by 57.71% ± 9.42%, 73.05% ± 11.82%, and 59.46% ± 9.91%, respectively. In addition, co-treatment reversed the phosphorylation level of AMP-activated protein kinase-α (AMPKα, which was reduced by OLZ, determined via the ratio of pAMPKα:AMPKα (94.1% compared with OLZ alone. The results showed that BBR may prevent lipid metabolism disorders caused by OLZ by reversing the degree of SREBP pathway upregulated and the phosphorylation of AMPKα downregulated. Collectively, these results indicated that BBR could be used as a potential adjuvant to prevent dyslipidemia and obesity caused by the use of second-generation antipsychotic medication.

  7. Derivation of a restriction map of bacteriophage T3 DNA and comparison with the map of bacteriophage T7 DNA.

    Science.gov (United States)

    Bailey, J N; Dembinski, D R; McAllister, W T

    1980-01-01

    The DNA of bacteriophage T3 was characterized by cleavage with seven restriction endonucleases. AvaI, XbaI, BglII, and HindIII each cut T3 DNA at 1 site, KpnI cleaved it at 2 sites, MboI cleaved it at 9 sites, and HpaI cleaved it at 17 sites. The sizes of the fragments produced by digestion with these enzymes were determined by using restriction fragments of T7 DNA as molecular weight standards. As a result of this analysis, the size of T3 DNA was estimated to be 38.74 kilobases. The fragments were ordered with respect to each other and to the genetic map to produce a restriction map of T3 DNA. The location and occurrence of the restriction sites in T3 DNA are compared with those in the DNA of the closely related bacteriophage T7. Images PMID:6251266

  8. Model remediation of contaminated sites as illustrated by the example of the TNT-remediation project Stadtallendorf, Hessen, Germany. Part 1: Final report; Part 2: Documentation and balancing of the measures; Part 3: Documentation of citizens' participation and public relations activities in Stadtallendorf; Modellhafte Sanierung von Altlasten am Beispiel des TNT-Sanierungsprojektes Stadtallendorf/Hessen. T. 1: Abschlussbericht. T. 2: Dokumentation und Bilanzierung von Auswirkungen der Sanierung. T. 3: Dokumentation der Buergerbeteiligung und Oeffentlichkeitsarbeit in Stadtallendorf

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    1999-07-01

    The project comprised the following stages: 1. Development of a technology for purification of soils contaminated with explosives. The process comprised soil washing, hot steam treatment, and thermal treatment of residues; 2. Projecting, construction and two years of operation of a soil purification plant with a rated throughput of 20 t/h and a total throughput of 130,000 t; 3. Remediation of two representative sites on DAG terrain (TNT production chain and filling stations); 4. Verification of the findings in an accompanying scientific programme. The site under investigation was formerly occupied by the Dynamit AG (DAG) Stadtallendorf, where TNT, contaminated waste water and acids were produced, and contamination levels were high in several areas. Today, the area is a residential area. In addition to the investigations described here, the waste water discharge system of the DAG was modernized. [German] Das FuE-Vorhaben Stadtallendorf wurde mit folgender Zielsetzung begonnen: (1) Entwicklung einer geeigneten und moeglichst umweltvertraeglichen Technik zur Reinigung von Boeden mit sprengstoffspezifischen Kontaminationen; hierzu wurde in Anlehnung an die Ausschreibung des Bundes ein kombiniertes Bodenbehandlungsverfahren vorgesehen, das aus folgenden Komponenten bestand: - Bodenwaesche, - Heissdampfbehandlung, - Thermische Reststoffbehandlung. (2) Planung, Bau und zweijaehriger Betrieb einer entsprechenden grosstechnischen Bodenreinigungsanlage (Nenndurchsatz von 20 t/Stunde; Gesamtdurchsatz 130.000 t). (3) Bodensanierung von zwei repraesentativen Arealen im DAG-Gelaende (TNT-Produktionskette und Fuellstellenbetrieb). (4) Absicherung der Ergebnisse in einem wissenschaftlichen Begleitprogramm. Gegenstand der Phase 2 des Vorhabens war der vollstaendige Prozess der Planung und Umsetzung der Sanierung eines (Teils eines) Planungsraumes des Ruestungsaltstandortes Dynamit AG (DAG) Stadtallendorf. Es handelte sich dabei um einen Bereich, der als Standort fuer TNT

  9. Characterization of VAMP isoforms in 3T3-L1 adipocytes: implications for GLUT4 trafficking.

    Science.gov (United States)

    Sadler, Jessica B A; Bryant, Nia J; Gould, Gwyn W

    2015-02-01

    The fusion of GLUT4-containing vesicles with the plasma membrane of adipocytes is a key facet of insulin action. This process is mediated by the formation of functional soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) complexes between the plasma membrane t-SNARE complex and the vesicle v-SNARE or VAMP. The t-SNARE complex consists of Syntaxin4 and SNAP23, and whereas many studies identify VAMP2 as the v-SNARE, others suggest that either VAMP3 or VAMP8 may also fulfil this role. Here we characterized the levels of expression, distribution, and association of all the VAMPs expressed in 3T3-L1 adipocytes to provide the first systematic analysis of all members of this protein family for any cell type. Despite our finding that all VAMP isoforms form SDS-resistant SNARE complexes with Syntaxin4/SNAP23 in vitro, a combination of levels of expression (which vary by >30-fold), subcellular distribution, and coimmunoprecipitation analyses lead us to propose that VAMP2 is the major v-SNARE involved in GLUT4 trafficking to the surface of 3T3-L1 adipocytes.

  10. TC10 is regulated by caveolin in 3T3-L1 adipocytes.

    Directory of Open Access Journals (Sweden)

    Dave Bridges

    Full Text Available BACKGROUND: TC10 is a small GTPase found in lipid raft microdomains of adipocytes. The protein undergoes activation in response to insulin, and plays a key role in the regulation of glucose uptake by the hormone. METHODOLOGY/PRINCIPAL FINDINGS: TC10 requires high concentrations of magnesium in order to stabilize guanine nucleotide binding. Kinetic analysis of this process revealed that magnesium acutely decreased the nucleotide release and exchange rates of TC10, suggesting that the G protein may behave as a rapidly exchanging, and therefore active protein in vivo. However, in adipocytes, the activity of TC10 is not constitutive, indicating that mechanisms must exist to maintain the G protein in a low activity state in untreated cells. Thus, we searched for proteins that might bind to and stabilize TC10 in the inactive state. We found that Caveolin interacts with TC10 only when GDP-bound and stabilizes GDP binding. Moreover, knockdown of Caveolin 1 in 3T3-L1 adipocytes increased the basal activity state of TC10. CONCLUSIONS/SIGNIFICANCE: Together these data suggest that TC10 is intrinsically active in vivo, but is maintained in the inactive state by binding to Caveolin 1 in 3T3-L1 adipocytes under basal conditions, permitting its activation by insulin.

  11. Digital Image Analysis of Reactive Oxygen Species and CA2+ in Mouse 3T3 Fibroblasts

    Science.gov (United States)

    Xu, Hongzhi; Liu, Dongwu; Chen, Zhiwei

    Recently, analysis of digital images with confocal microscope has become a routine technique and indispensable tool for cell biological studies and molecular investigations. Because the light emitted from the point out-of-focus is blocked by the pinhole and can not reach the detector, thus only an image of the fluorescence from the focal plane is imaged. In present studies, we use the probes 2', 7'-dichlorof luorescein diacetate (H2DCF-DA) and Fluo-3 AM to research reactive oxygen species (ROS) and Ca2+ in mouse 3T3 fibroblasts, respectively. Our results indicate that the distribution of ROS and Ca2+ were clearly seen in mouse 3T3 fibroblasts. Moreover, we acquired and quantified the fluorescence intensity of ROS and Ca2+ with Leica Confocal Software. It was found that the quantified fluorescence intensity of ROS and Ca2+ was 123.30.26±8.99 and 125.13±12.16, respectively. Taken together, our results indicate that it is a good method to research the distribution and fluorescence intensity of ROS and Ca2+ in cultured cells with confocal microscope.

  12. Pitting corrosion of Al2024-T3 in sodium chloride solution

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    Pitting corrosion behavior of Al2024-T3 in sodium chloride solution was investigated by using potentiodynamic scanning (PDS) measurements and electrochemical impedance spectroscopy (EIS) technique. When pitting corrosion of the alloy occurs, there exists a passive region in the anodic branch of PDS polarization curve, which is enlarged with the increasing of immersion time due to the competition of the halide ions with OH- ions to adsorb on the oxide film to form the corrosion products film and the increase of pitting corrosion area. Two capacitive semicircles were observed in complex plane plot. For more extensive pitting and general corrosion of Al2024-T3, the passive region in PDS disappeared, while another depressed semicircle was observed in Nyquist plot because of the formation of corrosion products film. On the other hand, the low frequency inductive loop, which had often been regarded as a manifestation of pitting or formation and precipitation of a salt film, was not observed, which indicates that the low frequency inductive loop can not be the characteristic of pitting corrosion or the formation of salt film. The results also show that higher reactant CPE exponent values will correspond to more extensive transformation of a metal surface by very localized corrosion, while general corrosion can result in a smaller CPE exponent value.

  13. Behavior of MC3T3-E1 Osteoblast Cultured on Chitosan Modified with Polyvinylpyrrolidone

    Institute of Scientific and Technical Information of China (English)

    XI Jing; GAO Yuan; KONG Lijun; GONG Yandao; ZHAO Nanming; ZHANG Xiufang

    2005-01-01

    The physical and chemical properties of four kinds of modified chitosan materials made by blending chitosan with polyvinylpyrrolidone (PVP) were investigated. All four of these modified chitosan materials were hydrophilic with water contact angles ranging from 59° to 69°. Fourier transform-infrared spectra of the modified materials showed a new band at 1288 cm-1, implying formation of a surface physical interpenetrating network structure. Enzyme linked immunosorbent assay results indicated that much less fibronectin was adsorbed on the modified materials than on only chitosan. The viability of MC3T3-E1 osteoblasts cultured on the materials was assessed by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl- 2H-tetrazolium bromide assay. The results show that adding PVP10000 into the chitosan promotes adhesion of MC3T3-E1 osteoblasts on the modified materials, but has no effect on cell growth and proliferation; while adding PVP40000 reduces cell adhesion, growth, and proliferation. The results suggest that the increased hydrophilicity of the material surface does not always improve its biocompatibility, which will influence the selection and design of biomaterials.

  14. Effects of salvia miltiorrhiza bunge (SMB) on MC3T3-E1 cells.

    Science.gov (United States)

    Ding, Y; Soma, S; Takano-Yamamoto, T; Matsumoto, S; Sakuda, M

    1995-12-01

    In previous research, we found that the Salvia Miltiorrhiza Bunge (SMB), a traditional Chinese medicinal herb, accelerates orthodontic tooth movement. In the present study, to characterize the actions of SMB on bone remodeling, we investigated the effect of SMB on DNA synthesis and alkaline phosphatase activity of murine osteoblast like cell-clone, MC3T3-E1 cells in vitro. Treatment of the cells with SMB for 72 hours caused significant increase in ALPase activity. It was found that SMB increased ALPase activity in a dose-dependent manner, and up to maximum at the concentration of 5.0 mg/ml. At that concentration, ALPase activity was about 135% greater than that of control. SMB at 5.0 mg/ml significantly stimulated ALPase activity of the cells in multilayer on day 8 or calcification stage on day 16, but inhibited it in the sparse stage on day 2 or subconfluency stage on day 4. SMB had no effect on DNA synthesis in any stage of culture. ALPase activity significantly increased at 48 hours and was up to 300% greater than that of control activity at 96 hours. These findings suggest that SMB directly stimulates ALPase activity of MC3T3-E1 cells in multilayer stage or calcification stage without any effect on proliferation. Locally administrated SMB may affect the differentiation of osteoblasts in vivo.

  15. Assessment of combined toxicity of heavy metals from industrial wastewaters on Photobacterium phosphoreum T3S

    Science.gov (United States)

    Zeb, BibiSaima; Ping, Zheng; Mahmood, Qaisar; Lin, Qiu; Pervez, Arshid; Irshad, Muhammad; Bilal, Muhammad; Bhatti, Zulfiqar Ahmad; Shaheen, Shahida

    2017-07-01

    This research work is focusing on the toxicities of heavy metals of industrial origin to anaerobic digestion of the industrial wastewater. Photobacterium phosphoreum T3S was used as an indicator organism. The acute toxicities of heavy metals on P. phosphoreum T3S were assessed during 15-min half inhibitory concentration (IC50) as indicator at pH 5.5-6. Toxicity assays involved the assessment of multicomponent mixtures using TU and MTI approaches. The results of individual toxicity indicated that the toxicity of Cd, Cu and Pb on P. phosphoreum increased with increasing concentrations and there was a linear correlation. The 15-min IC50 values of Cd, Cu and Pb were 0.537, 1.905 and 1.231 mg/L, respectively, and their toxic order was Cd > Pb > Cu. The combined effects of Cd, Cu and Pb were assayed by equivalent concentration mixing method. The results showed that the combined effects of Cd + Cu, Cd + Pb, Cu + Pb, Cd + Cu + Pb were antagonistic, antagonistic and partly additive. The combined effect of three heavy metals was partly additive.

  16. Cholinergic interneurons mediate fast VGluT3-dependent glutamatergic transmission in the striatum.

    Science.gov (United States)

    Higley, Michael J; Gittis, Aryn H; Oldenburg, Ian A; Balthasar, Nina; Seal, Rebecca P; Edwards, Robert H; Lowell, Bradford B; Kreitzer, Anatol C; Sabatini, Bernardo L

    2011-04-22

    The neurotransmitter glutamate is released by excitatory projection neurons throughout the brain. However, non-glutamatergic cells, including cholinergic and monoaminergic neurons, express markers that suggest that they are also capable of vesicular glutamate release. Striatal cholinergic interneurons (CINs) express the Type-3 vesicular glutamate transporter (VGluT3), although whether they form functional glutamatergic synapses is unclear. To examine this possibility, we utilized mice expressing Cre-recombinase under control of the endogenous choline acetyltransferase locus and conditionally expressed light-activated Channelrhodopsin2 in CINs. Optical stimulation evoked action potentials in CINs and produced postsynaptic responses in medium spiny neurons that were blocked by glutamate receptor antagonists. CIN-mediated glutamatergic responses exhibited a large contribution of NMDA-type glutamate receptors, distinguishing them from corticostriatal inputs. CIN-mediated glutamatergic responses were insensitive to antagonists of acetylcholine receptors and were not seen in mice lacking VGluT3. Our results indicate that CINs are capable of mediating fast glutamatergic transmission, suggesting a new role for these cells in regulating striatal activity.

  17. Cholinergic interneurons mediate fast VGluT3-dependent glutamatergic transmission in the striatum.

    Directory of Open Access Journals (Sweden)

    Michael J Higley

    Full Text Available The neurotransmitter glutamate is released by excitatory projection neurons throughout the brain. However, non-glutamatergic cells, including cholinergic and monoaminergic neurons, express markers that suggest that they are also capable of vesicular glutamate release. Striatal cholinergic interneurons (CINs express the Type-3 vesicular glutamate transporter (VGluT3, although whether they form functional glutamatergic synapses is unclear. To examine this possibility, we utilized mice expressing Cre-recombinase under control of the endogenous choline acetyltransferase locus and conditionally expressed light-activated Channelrhodopsin2 in CINs. Optical stimulation evoked action potentials in CINs and produced postsynaptic responses in medium spiny neurons that were blocked by glutamate receptor antagonists. CIN-mediated glutamatergic responses exhibited a large contribution of NMDA-type glutamate receptors, distinguishing them from corticostriatal inputs. CIN-mediated glutamatergic responses were insensitive to antagonists of acetylcholine receptors and were not seen in mice lacking VGluT3. Our results indicate that CINs are capable of mediating fast glutamatergic transmission, suggesting a new role for these cells in regulating striatal activity.

  18. Triiodothyronine (T3) effects on cardiovascular system in patients with heart failure.

    Science.gov (United States)

    Pingitore, Alessandro; Iervasi, Giorgio

    2008-01-01

    Interest in the role of thyroid hormones (TH) in heart failure is steadily increasing due to evidence for a physiological, homeostatic role of TH and the effects of altered TH metabolism on the cardiovascular system, particularly in presence of heart failure. Experimental studies have shown that altered TH metabolism modifies cardiovascular homeostasis by inducing alterations of cardiac histology, cardiomyocyte morphology and gene expression and consequently, of diastolic and systolic myocardial function. Clinical studies have shown that mild forms of thyroid dysfunction, both primary (subclinical hypothyroidism and subclinical hyperthyroidism) and secondary (low T(3) syndrome) have negative prognostic impact in patients with heart failure. In these patients, the administration of synthetic triiodothyronine (T(3)) was well tolerated and induced significant improvement in cardiac function without increased heart rate and metabolic demand. Large multicenter, placebo-controlled prospective studies are necessary to evaluate the safety and prognostic effects of chronic treatment with TH replacement therapy in patients with heart failure. The article also discusses recent patents in this field.

  19. The "T3 Support Centre" (Teaching, Technology and Testing - Not just another help desk

    Directory of Open Access Journals (Sweden)

    Carol Miles

    2005-10-01

    Full Text Available Many faculty members embrace the challenge of responding to rising student demands for more technically advanced course supports by offering their courseware through a variety of media. However, it is often difficult for them to find the time required to become proficient in the use of the software packages, course management systems and web technologies at their disposal. These new realities of teaching point to the need for support systems for faculty members that go beyond the traditional computer services "help desk" with a more comprehensive support service that actually becomes involved in the development and modification of technology-based course materials and computerized test marking and analysis. Increasing demand for these types of services at Carleton University resulted in the establishment of the T3 (Teaching...Technology...Testing Support Centre. The service offers faculty members extended-hour phone-in and walk-in support as well as a variety of resources such as Scantron and Item Analysis service for multiple choice exams, the use of scanners and colour printers, as well as a variety of teaching publications and contacts. This paper details the planning, administration, and services offered of the T3 Service, including advice those attempting to establish a similar service. Usage statistics from the first year of operations will be delineated.

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

    Science.gov (United States)

    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.

  1. Acanthamoeba genotypes T3 and T4 as causative agents of amoebic keratitis in Mexico.

    Science.gov (United States)

    Omaña-Molina, Maritza; Vanzzini-Zago, Virginia; Hernandez-Martinez, Dolores; Gonzalez-Robles, Arturo; Salazar-Villatoro, Lizbeth; Ramirez-Flores, Elizabeth; Oregon-Miranda, Eric; Lorenzo-Morales, Jacob; Martinez-Palomo, Adolfo

    2016-02-01

    Free-living amoebae (FLA) are widely distributed worldwide. Some genera included in this group act as opportunistic pathogens causing fatal encephalitis and Acanthamoeba keratitis (AK), a sight-threatening infection of the cornea associated with the use of soft contact lenses that could even end in blindness if an early diagnosis and treatment are not achieved. Furthermore, the numbers of AK cases keep rising worldwide mainly due to an increase of contact lens wearers and lack of hygiene in the maintenance of lenses and their cases. In Mexico, no cases of AK have been described so far although the isolation of other pathogenic FLA such as Naegleria fowleri and Balamuthia mandrillaris from both clinical and environmental sources has been reported. The present study reports two cases of Acanthamoeba keratitis diagnosed in two patients admitted to the Hospital "Luis Sánchez Bulnes" for Blindness Prevention in Mexico City, Mexico. Corneal scrapes and contact lenses were checked for the presence of Acanthamoeba strains in both patients. Strains were axenized after initial isolation to classify at the genotype level. After sequencing the diagnostic fragment 3 (DF3) region located on the 18S ribosomal DNA (rDNA) gene of Acanthamoeba, genotype T3 and genotype T4 were identified in clinical case 1 and 2, respectively. To our knowledge, these are the first reported cases of AK in Mexico in the literature and the first description of Acanthamoeba genotypes T3 and T4 as causative agents of amoebic infection.

  2. Agarose Gel Electrophoresis Reveals Structural Fluidity of a Phage T3 DNA Packaging Intermediate

    Science.gov (United States)

    Serwer, Philip; Wright, Elena T.

    2012-01-01

    We find a new aspect of DNA packaging-associated structural fluidity for phage T3 capsids. The procedure is (1) glutaraldehyde cross-linking of in vivo DNA packaging intermediates for stabilization of structure and then (2) determining of effective radius by two-dimensional agarose gel electrophoresis (2d-AGE). The intermediates are capsids with incompletely packaged DNA (ipDNA) and without an external DNA segment; these intermediates are called ipDNA-capsids. We initially increase production of ipDNA-capsids by raising NaCl concentration during in vivo DNA packaging. By 2d-AGE, we find a new state of contracted shell for some particles of one previously identified ipDNA-capsid. The contracted shell-state is found when ipDNA length/mature DNA length (F) is above 0.17, but not at lower F. Some contracted-shell ipDNA-capsids have the phage tail; others do not. The contracted-shell ipDNA-capsids are explained by premature DNA maturation cleavage that makes accessible a contracted-shell intermediate of a cycle of the T3 DNA packaging motor. The analysis of ipDNA-capsids, rather than intermediates with uncleaved DNA, provides a simplifying strategy for a complete biochemical analysis of in vivo DNA packaging. PMID:22222979

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

    Science.gov (United States)

    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. Copyright © 2016 Elsevier Ltd. All rights reserved.

  4. Wogonin enhances intracellular adiponectin levels and suppresses adiponectin secretion in 3T3-L1 adipocytes.

    Science.gov (United States)

    Yang, Tan; Liu, Hua; Zhao, Bo; Xia, Zhongyuan; Zhang, Yemin; Zhang, Deling; Li, Mingxin; Cao, Yingkang; Zhang, Zhijiang; Bi, Yongyi; Wang, Changhua

    2017-01-30

    As an insulin sensitizer and modulator of inflammatory responses, adiponectin has become a therapeutic target for insulin resistance, diabetes, and diabetes-related complications. Wogonin possesses anti-oxidative, anti-inflammatory, and anti-diabetic abilities. However, its effect on generation and secretion of adiponectin is ill-defined in adipocytes. Here, we demonstrated that wogonin administration augmented intracellular adiponectin levels and attenuated adiponectin release in a dose- and time-dependent manner in mature 3T3-L1 adipocytes, along with a suppression of PKCδ phosphorylation. Wogonin treatment also prevented PKCδ overexpression-induced reduction of intracellular adiponectin levels and enhancement of adiponectin release. In addition, wogonin supplementation dramatically increased AMPK phosphorylation and SirT1 expression. Inhibition of either AMPK or SirT1 mitigated wogonin action on adiponectin production and release. Furthermore, inhibition of AMPK by its specific inhibitor markedly reduced wogonin-enhanced mRNA and protein expressions of SirT1. These results suggested that wogonin regulated expression and secretion of adiponectin via PKCδ/AMPK/SirT1 signaling pathway in mature 3T3-L1 adipocytes.

  5. Combination therapy with catechins and caffeine inhibits fat accumulation in 3T3-L1 cells

    Science.gov (United States)

    Zhu, Xiaojuan; Yang, Licong; Xu, Feng; Lin, Lezhen; Zheng, Guodong

    2017-01-01

    Catechins and caffeine, which are green tea components, have a slimming effect; however, the combinational effect of fat metabolism in 3T3-L1 cells remains unclear. In the present study, 3T3-L1 cells were treated with catechins and caffeine in combination, and it was found that combination therapy with catechins and caffeine markedly reduced intracellular fat accumulation, mRNA expression levels of peroxisome proliferator-activated receptor-γ and CCAAT/enhancer-binding protein α in the early stage of cell differentiation were significantly reduced, and mRNA expression of fatty acid synthetase(FAS) andglycerol-3-phosphate dehydrogenase protein expression levels of FAS were downregulated. Noradrenaline-induced lipolysis was enhanced by caffeine, which markedly increased the protein expression of adipose triglyceride lipase and hormone sensitive lipase. These results indicated that combination therapy with catechins and caffeine synergistically inhibited lipid accumulation by regulating the gene and protein expression levels of lipid metabolism-related enzymes. Therefore, catechins and caffeine combination therapy has potential as a functional food that may be used to prevent obesity and lifestyle-associated diseases. PMID:28352352

  6. Protein kinase A suppresses the differentiation of 3T3-L1 preadipocytes

    Institute of Scientific and Technical Information of China (English)

    Fuqiang Li; Dongmei Wang; Yiran Zhou; Bo Zhou; Yanan Yang; Hehua Chen; Jianguo Song

    2008-01-01

    cAMP and protein kinase A (PKA) are widely known as signaling molecules that are important for the induction of adipogenesis. Here we show that a strong increase in the amount of cAMP inhibits the adipogenesis of 3T3-L1 fibroblast cells. Stimulation of PKA activity suppresses adipogenesis and, in contrast, inhibition of PKA activity markedly accelerates the adipogenic process. As adipogenesis progresses, there is a significant increase in the expression level of PKA regulatory subunits and a corresponding decrease in PKA activity. Moreover, treatment of 3T3-L1 cells with epidermal growth factor (EGF) stimulates PKA activity and blocks adipogenesis. Inhibition of PKA activity abolishes this suppressive effect of EGF on adipogenesis. Moreover, activation of PKA induces serine/threonine phosphorylation, reduces tyrosine phosphorylation of insulin receptor substrate 1 (IRS-1) and the association between PKA and IRS-1. Taken together, our study demonstrates that PKA has a pivotal role in the suppression of adipogenesis. cAMP at high concentrations can suppress adipogenesis through PKA activation. These findings could be important and useful for understanding the mechanisms of adipogenesis and the relevant physiological events.

  7. The "T3 Support Centre" (Teaching, Technology and Testing - Not just another help desk

    Directory of Open Access Journals (Sweden)

    Carol Miles

    2005-10-01

    Full Text Available Many faculty members embrace the challenge of responding to rising student demands for more technically advanced course supports by offering their courseware through a variety of media. However, it is often difficult for them to find the time required to become proficient in the use of the software packages, course management systems and web technologies at their disposal. These new realities of teaching point to the need for support systems for faculty members that go beyond the traditional computer services "help desk" with a more comprehensive support service that actually becomes involved in the development and modification of technology-based course materials and computerized test marking and analysis. Increasing demand for these types of services at Carleton University resulted in the establishment of the T3 (Teaching...Technology...Testing Support Centre. The service offers faculty members extended-hour phone-in and walk-in support as well as a variety of resources such as Scantron and Item Analysis service for multiple choice exams, the use of scanners and colour printers, as well as a variety of teaching publications and contacts. This paper details the planning, administration, and services offered of the T3 Service, including advice those attempting to establish a similar service. Usage statistics from the first year of operations will be delineated.

  8. Characterization of Catalase from Psychrotolerant Psychrobacter piscatorii T-3 Exhibiting High Catalase Activity

    Directory of Open Access Journals (Sweden)

    Hidetoshi Matsuyma

    2012-02-01

    Full Text Available A psychrotolerant bacterium, strain T-3 (identified as Psychrobacter piscatorii, that exhibited an extraordinarily high catalase activity was isolated from the drain pool of a plant that uses H2O2 as a bleaching agent. Its cell extract exhibited a catalase activity (19,700 U·mg protein−1 that was higher than that of Micrococcus luteus used for industrial catalase production. Catalase was approximately 10% of the total proteins in the cell extract of the strain. The catalase (PktA was purified homogeneously by only two purification steps, anion exchange and hydrophobic chromatographies. The purified catalase exhibited higher catalytic efficiency and higher sensitivity of activity at high temperatures than M. luteus catalase. The deduced amino acid sequence showed the highest homology with catalase of Psycrobacter cryohalolentis, a psychrotolelant bacterium obtained from Siberian permafrost. These findings suggest that the characteristics of the PktA molecule reflected the taxonomic relationship of the isolate as well as the environmental conditions (low temperatures and high concentrations of H2O2 under which the bacterium survives. Strain T-3 efficiently produces a catalase (PktA at a higher rate than Exiguobacterium oxidotolerans, which produces a very strong activity of catalase (EktA at a moderate rate, in order to adapt to high concentration of H2O2.

  9. Anti-adipogenic effect of mulberry leaf ethanol extract in 3T3-L1 adipocytes.

    Science.gov (United States)

    Yang, Soo Jin; Park, Na-Young; Lim, Yunsook

    2014-12-01

    Adipogenesis is part of the cell differentiation process in which undifferentiated fibroblasts (pre-adipocytes) become mature adipocytes with the accumulation of lipid droplets and subsequent cell morphological changes. Several transcription factors and food components have been suggested to be involved in adipogenesis. The aim of this study was to determine whether mulberry leaf ethanol extract (MLEE) affects adipogenesis in 3T3-L1 adipocytes. The 3T3-L1 adipocytes were treated with different doses of MLEE for 8 days starting 2 days post-confluence. Cell viability, fat accumulation, and adipogenesis-related factors including CCAAT-enhancer-binding protein alpha (C/EBPα), peroxisome proliferator-activated receptor gamma (PPARγ), PPARγ coactivator 1 alpha (PGC-1α), fatty acid synthase (FAS), and adiponectin were analyzed. Results showed that MLEE treatments at 10, 25, 50, and 100 µg/ml had no effect on cell morphology and viability. Without evident toxicity, all MLEE treated cells had lower fat accumulation compared with control as shown by lower absorbances of Oil Red O stain. MLEE at 50 and 100 µg/ml significantly reduced protein levels of PPARγ, PGC-1α, FAS, and adiponectin in differentiated adipocytes. Furthermore, protein level of C/EBPα was significantly decreased by the treatment of 100 µg/ml MLEE. These results demonstrate that MLEE treatment has an anti-adipogenic effect in differentiated adipocytes without toxicity, suggesting its potential as an anti-obesity therapeutic.

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

    Science.gov (United States)

    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.

  11. Left ventricular function by speckle-tracking echocardiography in patients with low-T3 syndrome and acute myocardial infarction.

    Science.gov (United States)

    Jankauskienė, Edita; Orda, Paulius; Rumbinaitė, Eglė; Žaliaduonytė-Pekšienė, Diana; Steponavičiutė, Rasa; Krasauskienė, Aurelija; Vaškelytė, Jolanta Justina; Bunevičius, Robertas

    2015-01-01

    Low-T3 syndrome is common in patients with acute myocardial infarction (AMI). Recent experimental and clinical data have suggested a potential negative impact of low-T3 syndrome on myocardial function in patients with AMI. The aim of this study was to assess left ventricular (LV) myocardial function in patients with low-T3 syndrome and to investigate the association between hormonal profile and the severity of LV dysfunction using speckle-tracking echocardiography (STE). In 130 patients with first-onset ST-segment elevation acute myocardial infarction (STEMI), conventional 2D and speckle-tracking echocardiography within 48-72h after the hospitalization was performed, and blood samples for TSH, fT4, fT3, and anti-TPO levels were obtained to investigate thyroid hormone production within 24h and on the fourth day after the onset of STEMI symptoms. The patients were divided into two groups according to their serum level of fT3: group 1 with fT3 concentration below 3.2pmol/L (n=34) and group 2 with normal fT3 (>3.2pmol/L) level (n=96). LV ejection fraction (EF) tended to be lower in the low fT3 group. The systolic longitudinal strain did not differ between the groups, but the late diastolic longitudinal strain rate was lower in group 1 (P=0.011). The systolic basal LV rotation positively correlated with the level of fT3 (r=0.4; Pspeckle-tracking echocardiography were impaired in patients with low-T3 syndrome after AMI. Copyright © 2015 Lithuanian University of Health Sciences. Production and hosting by Elsevier Urban & Partner Sp. z o.o. All rights reserved.

  12. Lack of action of exogenously administered T3 on the fetal rat brain despite expression of the monocarboxylate transporter 8.

    Science.gov (United States)

    Grijota-Martínez, Carmen; Díez, Diego; Morreale de Escobar, Gabriella; Bernal, Juan; Morte, Beatriz

    2011-04-01

    Mutations of the monocarboxylate transporter 8 gene (MCT8, SLC16A2) cause the Allan-Herndon-Dudley syndrome, an X-linked syndrome of severe intellectual deficit and neurological impairment. Mct8 transports thyroid hormones (T4 and T3), and the Allan-Herndon-Dudley syndrome is likely caused by lack of T3 transport to neurons during critical periods of fetal brain development. To evaluate the role of Mct8 in thyroid hormone action in the fetal brain we administered T4 or T3 to thyroidectomized pregnant dams treated with methyl-mercapto-imidazol to produce maternal and fetal hypothyroidism. Gene expression was then measured in the fetal cerebral cortex. T4 increased Camk4, Sema3c, and Slc7a3 expression, but T3 was without effect. To investigate the cause for the lack of T3 action we analyzed the expression of organic anion transport polypeptide (Oatp14, Slco1c1), a T4 transporter, and Mct8 (Slc16a2), a T4 and T3 transporter, by confocal microscopy. Both proteins were present in the brain capillaries forming the blood-brain barrier and in the epithelial cells of the choroid plexus forming the blood-cerebrospinal fluid barrier. It is concluded that T4 from the maternal compartment influences gene expression in the fetal cerebral cortex, possibly after transport via organic anion transporter polypeptide and/or Mct8, and conversion to T3 in the astrocytes. On the other hand, T3 does not reach the target neurons despite the presence of Mct8. The data indicate that T4, through local deiodination, provides most T3 in the fetal rat brain. The role of Mct8 as a T3 transporter in the fetal rat brain is therefore uncertain.

  13. 利培酮抑制3T3-L1前脂肪细胞的分化%Risperidone inhibits 3T3-L1 pre-adipocytes differentiation

    Institute of Scientific and Technical Information of China (English)

    张高丽; 张弋; 于海川; 张新雅

    2016-01-01

    Objective To investigate the influence of risperidone on differentiation of 3T3‐L1 pre‐adipocytes .Methods 3T3‐L1 pre‐adipocytes were induced to differentiate into mature adipocytes by adopting the classic hormone cocktail method and observed by the oil red O staining .Meanwhile ,the inducing medium was added with risperidone for studying its influence on 3T3‐L1 pre‐adi‐pocytes differentiation .Results 3T3‐L1 pre‐adipocytes were successfully differentiated into the mature adipocytes ,0 .1 ,1 ,10μmol/L risperidone all could inhibit the differentiation of 3T3‐L1 pre‐adipocytes .Conclusion Risperidone can inhibit the differentiation of 3T3‐L1 pre‐adipocytes .%目的:研究利培酮对3T3‐L1前脂肪细胞分化的影响。方法采用经典的激素鸡尾酒法诱导3T3‐L1前脂肪细胞分化为成熟的脂肪细胞,油红O染色观察。向诱导培养基中加入利培酮研究其对3T3‐L1前脂肪细胞分化的影响。结果用激素鸡尾酒法成功地将3T3‐L1前脂肪细胞诱导为成熟的脂肪细胞。0.1、1、10μmol/L的利培酮均能够抑制3T3‐L1前脂肪细胞的分化。结论利培酮能够抑制3T3‐L1前脂肪细胞的分化。

  14. 小檗碱对白介素-6诱导胰岛素抵抗3T3-L1脂肪细胞脂联素表达的影响%Effect of Berberine on Gene Expression of Adiponectin by IL-6 Induced Insulin Resistance in 3T3-L1 Adipocyte

    Institute of Scientific and Technical Information of China (English)

    章常华; 李冰涛; 屈飞; 魏学鑫; 余日跃; 徐国良

    2012-01-01

    Objective; To explore the effect of berberine on adiponectin gene mRNA expression in 3T3-Ll adipocyte of insulin resistant induced by interleukin-6 (IL-6). Method; To build 3T3-L1 adipocyte model of insulin resistant induced by interleukin-6. 3T3-L1 adipocytes were exposed to 20 μg -L-1 IL-6 for 48 h, the 3T3-L1 adipocytes with insulin resistant were divided into six groups randomly; normal control groups, model groups, pioglitazone (50 (μmol -L-1 ) , and low, middle, hight doses of berberine groups (10, 20, 50 (μmol -L-1) , the amount of glucose consumption was measured by detecting the glucose content in cell culture supernatants with glucose oxidase assay, the effect of berberine on glucose uptake of 3T3-L1 adipocytes was observed, and cell model of insulin resistant was identified. mRNA expression of adiponection gene was determined by real-time quantitative reverse-transcription polymerase chain reaction. Result; There was statistical difference between model group and normal control group in their amount of glucose consumption and gene expression of adiponectin, which was decreased markedly in model group (P < 0. 05 ). Pioglitazone (50 (μmol -L-1 ) , and low, middle, high doses of berberine groups (10, 20, 50 (μmol -L-1 ) markedly increased both amount of glucose consumption and gene expression of adiponectin. Conclusion; Berberine can increase mRNA expression of adiponection gene and improve insulin resistant in 3T3-L1 adipocytes.%目的:观察小檗碱对白介素-6(IL-6)诱导胰岛素抵抗的3T3-L1脂肪细胞脂联素表达的影响.方法:选用IL-6诱导3T3-L1脂肪细胞胰岛素抵抗(IR)模型.以20 μg·L-1IL-6培养48 h,将3T3-L1脂肪细胞随机分为正常对照组、模型组、吡格列酮组(50μmol·L-1)和小檗碱高、中、低剂量组(10,20,50 μmol·L-1),以葡萄糖氧化酶法测定葡萄糖消耗量,观察小檗碱对脂肪细胞葡萄糖摄取的影响,鉴定IR模型;采用实时荧光定量PCR技术测定脂肪细胞脂联

  15. Effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin on adipogenic differentiation and insulin-induced glucose uptake in 3T3-L1 cells

    Energy Technology Data Exchange (ETDEWEB)

    Hsu, Hsin-Fen [Division of Environmental Health and Occupational Medicine, National Health Research Institutes, Zhunan, Miaoli County 35053, Taiwan (China); Tsou, Tsui-Chun, E-mail: tctsou@nhri.org.tw [Division of Environmental Health and Occupational Medicine, National Health Research Institutes, Zhunan, Miaoli County 35053, Taiwan (China); Chao, How-Ran [Department of Environmental Science and Engineering, National Pingtung University of Science and Technology, Neipu 912, Pingtung, Taiwan (China); Kuo, Ya-Ting; Tsai, Feng-Yuan; Yeh, Szu-Ching [Division of Environmental Health and Occupational Medicine, National Health Research Institutes, Zhunan, Miaoli County 35053, Taiwan (China)

    2010-10-15

    Dioxin exposure has been positively associated with human type II diabetes. Because lipophilic dioxins accumulate mainly in adipose tissue, this study aimed to determine if dioxins induce metabolic dysfunction in fat cells. Using 3T3-L1 cells as an in vitro model, we analyzed the effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), a model dioxin, on adipogenic differentiation, glucose uptake, and lipolysis. TCDD inhibited adipogenic differentiation, as determined by using oil droplet formation and adipogenic marker gene expression, including PPAR{gamma} (peroxisome proliferator-activated receptor {gamma}), C/EBP{alpha} (CCAAT/enhancer-binding protein {alpha}), and Glut4 (glucose transporter type 4). Effects of TCDD on glucose uptake were evaluated using fully differentiated 3T3-L1 adipocytes, revealing that TCDD significantly attenuated insulin-induced glucose uptake dose dependently. Inhibition of aryl hydrocarbon receptor (AhR) by {alpha}-naphthoflavone ({alpha}-NF), an AhR inhibitor, did not prevent the inhibitory effect of TCDD on glucose uptake, suggesting that TCDD attenuates insulin-induced glucose uptake in an AhR-independent manner. Effects of TCDD on lipolysis were determined using glycerol release assay. We found that TCDD had no marked effect on isoproterenol-induced glycerol release in fully differentiated 3T3-L1 adipocytes. These results provide in vitro evidence of TCDD's effects on fat cell metabolism, suggesting dioxin exposure in development of insulin resistance and type II diabetes.

  16. Resveratrol potentiates genistein's antiadipogenic and proapoptotic effects in 3T3-L1 adipocytes.

    Science.gov (United States)

    Rayalam, Srujana; Della-Fera, Mary Anne; Yang, Jeong-Yeh; Park, Hea Jin; Ambati, Suresh; Baile, Clifton A

    2007-12-01

    Genistein (G) and resveratrol (R) individually inhibit adipogenesis in 3T3-L1 adipocytes and induce apoptosis in cancer cells. We investigated whether the combination of G and R resulted in enhanced effects on adipogenesis, lipolysis, and apoptosis in 3T3-L1 cells. Preadipocytes and mature adipocytes were treated with G and R individually at 50 and 100 micromol/L (G100; R100) and in combination. Both in preadipocytes and mature adipocytes, G and R individually decreased cell viability dose-dependently, but G100 + R100 further decreased viability by 59 +/- 0.97% (P < 0.001) and 69.7 +/- 1.2% (P < 0.001) after 48 h compared with G100 and R100, respectively. G100 + R100 induced apoptosis 242 +/- 8.7% (P < 0.001) more than the control after 48 h, whereas G100 and R100 individually increased apoptosis only 46 +/- 9.2 and 46 +/- 7.9%, respectively. G and R did not modulate mitogen-activated protein kinase expression by themselves, but G100 + R100 increased Jun-N-terminal kinase phosphorylation by 38.8 +/- 4.4% (P < 0.001) and decreased extracellular signal-regulating kinase phosphorylation by 48 +/- 3.4% (P < 0.001). Individually, G and R at 25 micromol/L (G25; R25) decreased lipid accumulation by 30 +/- 1.7% and 20.07 +/- 4.27%, respectively (P < 0.001). However, G25 + R25 decreased lipid accumulation by 77.9 +/- 3.4% (P < 0.001). Lipolysis assay revealed that neither G25 nor R25 induced lipolysis, whereas G25 + R25 significantly increased lipolysis by 25.5 +/- 4.6%. The adipocyte-specific proteins PPARgamma and CCAAT/enhancer binding protein-alpha were downregulated after treatment with G + R, but no effect was observed with individual compounds. These results indicate that G and R in combination produce enhanced effects on inhibiting adipogenesis, inducing apoptosis, and promoting lipolysis in 3T3-L1 adipocytes. Thus, the combination of G and R is more potent in exerting antiobesity effects than the individual compounds.

  17. Ursolic acid inhibits adipogenesis in 3T3-L1 adipocytes through LKB1/AMPK pathway.

    Directory of Open Access Journals (Sweden)

    Yonghan He

    Full Text Available 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

  18. Effects of T3 treatment on brown adipose tissue and energy expenditure in a patient with craniopharyngioma and hypothalamic obesity.

    Science.gov (United States)

    van Santen, Hanneke M; Schouten-Meeteren, Antoinette Y; Serlie, Mireille; Meijneke, Ruud W H; van Trotsenburg, A S; Verberne, Hein; Holleman, Frits; Fliers, Eric

    2015-01-01

    Patients treated for childhood craniopharyngioma often develop hypothalamic obesity (HO), which has a huge impact on the physical condition and quality of life of these patients. Treatment for HO thus far has been disappointing, and although several different strategies have been attempted, all interventions had only transient effects. Since thyroid hormones increase energy expenditure metabolism (thyroid hormone induced thermogenesis), it was speculated that treatment with tri-iodothyronine (T3) may be beneficial. In 2002, a case report was published on reduction of body weight after T3 treatment for HO. No studies have been reported since. Recent experimental studies in rodents showed that T3 increases brown adipose tissue (BAT) activity via (pre)sympathetic pathways between the hypothalamus and BAT. Our aim was to investigate whether T3 treatment increases BAT activity in a patient with HO resulting from (treatment of) childhood craniopharyngioma. Thyroxine treatment for central hypothyroidism was switched to T3 monotherapy. Serum T3 and free thyroxine (FT4) concentrations were measured twice weekly for 2 months. ¹²³I-MIBG and ¹⁸F-FDG-PET after induction of non-shivering thermogenesis for the assessment of sympathetic and metabolic activity of BAT as well as indirect calorimetry for assessment of resting energy expenditure were performed before and during T3 treatment. No change in sympathetic and metabolic BAT activity, energy expenditure, or BMI was seen during T3 treatment despite the expected changes in thyroid hormone plasma concentrations. We conclude that T3 monotherapy does not seem to be effective in decreasing HO in childhood craniopharyngioma.

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

    Science.gov (United States)

    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; Alkema, Wynand

    2013-05-01

    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.

  20. Cell shrinkage as a signal to apoptosis in NIH 3T3 fibroblasts

    DEFF Research Database (Denmark)

    Friis, Martin B; Friborg, Christel R; Schneider, Linda

    2005-01-01

    Cell shrinkage is a hallmark of the apoptotic mode of programmed cell death, but it is as yet unclear whether a reduction in cell volume is a primary activation signal of apoptosis. Here we studied the effect of an acute elevation of osmolarity (NaCl or sucrose additions, final osmolarity 687...... mosmol l(-1)) on NIH 3T3 fibroblasts to identify components involved in the signal transduction from shrinkage to apoptosis. After 1.5 h the activity of caspase-3 started to increase followed after 3 h by the appearance of many apoptotic-like bodies. The caspase-3 activity increase was greatly enhanced...... in cells expressing a constitutively active G protein, Rac (RacV12A3 cell), indicating that Rac acts upstream to caspase-3 activation. The stress-activated protein kinase, p38, was significantly activated by phosphorylation within 30 min after induction of osmotic shrinkage, the phosphorylation being...

  1. Tissue transglutaminase expression affects hypusine metabolism in BALB/c 3T3 cells.

    Science.gov (United States)

    Beninati, S; Gentile, V; Caraglia, M; Lentini, A; Tagliaferri, P; Abbruzzese, A

    1998-10-16

    Post-translational formation of hypusine in eukaryotic initiation factor 5A (eIF-5A) is essential for cell viability. Recently, we showed that hypusine protein is an in vitro substrate for transglutaminases (TGases). We report the effect of tissue TGase expression on the in vivo hypusine metabolic pathway. The stable expression of tTGase in BALB/c 3T3 cells induced a 100-fold reduction of hypusine levels and a 50% increase of gamma-glutamyl-omega-hypusine formation. Such changes were paralleled by a consistent decrease in the free polyamine pool and an enhancement of their excretion and of the formation of their gamma-glutamyl derivatives. These effects occurred together with a significant reduction of cell proliferation. In this report we suggest, for the first time, that tTGase affects hypusine metabolism, thus regulating the eIF-5A activity and cell proliferation.

  2. Irradiation of Polystyrene and Polypropylene to study NIH 3T3 fibroblasts adhesion

    Energy Technology Data Exchange (ETDEWEB)

    Arbeitman, C.R. [Gerencia de Investigacion y Aplicaciones, TANDAR-CNEA (Argentina); Grosso, M.F. del, E-mail: delgrosso@tandar.cnea.gov.a [Gerencia de Investigacion y Aplicaciones, TANDAR-CNEA (Argentina); Consejo Nacional de Investigaciones Cientificas y Tecnicas, CONICET (Argentina); Ibanez, I. [Consejo Nacional de Investigaciones Cientificas y Tecnicas, CONICET (Argentina); Gerencia de Aplicaciones Tecnologicas de la Energia Nuclear, Dpto. de Radiobiologia, TANDAR-CNEA (Argentina); Garcia Bermudez, G. [Gerencia de Investigacion y Aplicaciones, TANDAR-CNEA (Argentina); Consejo Nacional de Investigaciones Cientificas y Tecnicas, CONICET (Argentina); Escuela de Ciencia y Tecnologia, UNSAM (Argentina); Duran, H. [Consejo Nacional de Investigaciones Cientificas y Tecnicas, CONICET (Argentina); Gerencia de Aplicaciones Tecnologicas de la Energia Nuclear, Dpto. de Radiobiologia, TANDAR-CNEA (Argentina); Escuela de Ciencia y Tecnologia, UNSAM (Argentina); Chappa, V.C. [Gerencia de Investigacion y Aplicaciones, TANDAR-CNEA (Argentina); Consejo Nacional de Investigaciones Cientificas y Tecnicas, CONICET (Argentina); Mazzei, R. [U.A. Tecnologicas y Agropecuarias, CNEA, Dpto. Ing. Quimica, UTN FRBA, Bs. As. (Argentina); Behar, M. [Instituto de Fisica, Universidade Federal do Rio Grande do Sul, Porto Alegre (Brazil)

    2010-10-01

    When polymers are irradiated with heavy ions new chemical groups are created in a few microns of the material. The irradiation changed the polarity and wettability on the surface so that could enhance the biocompatibility of the modified polymer. The study of chemistry and nanoscale topography of the biomaterial is important in determining its potential applications in medicine and biotechnology, because their strong influence on cell function, adhesion and proliferation. In this study, thin films of Polystyrene and Polypropylene samples were modified by irradiation with low energy ion beams (30-150 keV) and swift heavy ions both with various fluences and energies. The changes were evaluated with different methods. Adhesion of NIH 3T3 fibroblasts onto unirradiated and irradiated surfaces has been studied by in vitro techniques. The correlations between physicochemical properties as a function of different irradiations parameters were compared with cell adhesion on the modified polymer surface.

  3. Fructose promotes the differentiation of 3T3-L1 adipocytes and accelerates lipid metabolism.

    Science.gov (United States)

    Legeza, Balázs; Balázs, Zoltán; Odermatt, Alex

    2014-01-31

    Excessive fructose consumption and elevated glucocorticoids contribute to metabolic syndrome. We show that fructose as the only carbohydrate source is sufficient for the differentiation of 3T3-L1 fibroblasts into adipocytes. Differentiation of cells in fructose containing medium resulted in increased 11β-hydroxysteroid dehydrogenase 1 (11β-HSD1) expression and activity. Experiments with transfected HEK-293 cells suggested more efficient NADPH generation by fructose compared with glucose in the endoplasmic reticulum (ER). Adipocytes differentiated in the presence of fructose showed increased FABP4 expression, C/EBPα to C/EBPβ ratio and lipolysis. Thus, excessive fructose may cause adverse metabolic effects by enhancing 11β-HSD1 activity and increasing lipolysis in adipocytes. Copyright © 2014 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.

  4. Friction stir spot welding of 2024-T3 aluminum alloy with SiC nanoparticles

    Energy Technology Data Exchange (ETDEWEB)

    Paidar, Moslem; Sarab, Mahsa Laali [Islamic Azad University, Tehran (Iran, Islamic Republic of)

    2016-01-15

    In this study, the Friction stir spot welding (FSSW) of 2024-T3 aluminum alloy with 1.6 mm thickness was investigated. The effects of the silicon carbide (SiC) nanoparticles on the metallurgical and mechanical properties were discussed. The effects of particles on tension shear and wear tests were also investigated. The process was conducted at a constant rotational speed of 1000 rpm. Results showed that adding SiC nanoparticles to the weld during FSSW had a major effect on the mechanical properties. In fact, the addition of nanoparticles as barriers prevented grain growth in the Stir zone (SZ). The data obtained in the tensile-shear and wear tests showed that tensile-shear load and wear resistance increased with the addition of SiC nanoparticles, which was attributed to the fine grain size produced in the SZ.

  5. Pinless Friction Stir Welding of AA2024-T3 Joint and Its Failure Modes

    Institute of Scientific and Technical Information of China (English)

    李文亚; 李锦锋; 张志函; 高大路; 王卫兵; 栾国红

    2014-01-01

    The joining of aluminum alloy sheets with thickness less than 2.0 mm is difficult via conventional friction stir welding owing to the defects in the joint, such as root flaw, keyhole and lazy S. In the present research, a newly designed pinless tool with involute grooves on its shoulder surface was applied to weld 1.5 mm thick AA2024-T3. The effects of the rotating speed and welding speed on the microstructure and mechanical properties of the joints were ana-lyzed. The experimental results showed that the root flaw and keyhole were successfully eliminated. The lazy S was also eliminated under the optimized welding parameters. The maximum tensile strength of the joints was 326 MPa, which is about 74.1% that of the base material. Moreover, all the tensile samples fractured from the retreating side. Two fracture modes were observed during the tensile tests, which are related with the lazy S.

  6. Six new chalcones from Angelica keiskei inducing adiponectin production in 3T3-L1 adipocytes.

    Science.gov (United States)

    Ohnogi, Hiromu; Kudo, Yoko; Tahara, Kenichi; Sugiyama, Katsumi; Enoki, Tatsuji; Hayami, Shoko; Sagawa, Hiroaki; Tanimura, Yuko; Aoi, Wataru; Naito, Yuji; Kato, Ikunoshin; Yoshikawa, Toshikazu

    2012-01-01

    Angelica keiskei (Ashitaba in Japanese), a traditional herb in Japan, contains abundant prenylated chalcones. It has been reported that the chalcones from A. keiskei showed such bioactivities as anti-bacterial, anti-cancer and anti-diabetic effects. Xanthoangelol, 4-hydroxyderricin and six new chalcones were isolated in this study from an ethanol extract of A. keiskei by octadecyl silyl (ODS) and silica gel chromatography, and identified by 1D- and 2D-nuclear magnetic resonance (NMR) and high-resolution mass spectrometric analyses. The chalcones from A. keiskei markedly increased the expression of the adiponectin gene and the production of adiponectin in 3T3-L1 adipocytes. These results suggest that the chalcones from A. keiskei might be useful for preventing the metabolic syndrome.

  7. Hormone and pharmaceutical regulation of ASP production in 3T3-L1 adipocytes.

    Science.gov (United States)

    Gao, Ying; Gauvreau, Danny; Cianflone, Katherine

    2010-04-01

    Several studies have demonstrated increases in acylation stimulating protein (ASP), and precursor protein C3 in obesity, diabetes and dyslipidemia, however the nature of the regulation is unknown. To evaluate chronic hormonal and pharmaceutical mediated changes in ASP and potential mechanisms, 3T3-L1 adipocytes were treated with physiological concentrations of relevant hormones and drugs currently used in treatment of metabolic diseases for 48 h. Medium ASP production and C3 secretion were evaluated in relation to changes in adipocyte lipid metabolism (cellular triglyceride (TG) mass, non-esterified fatty acid (NEFA) release and real-time FA uptake). Chylomicrons increased ASP production (up to 411 +/- 133% P ASP production (-53 to -85%, P ASP (-54% to -100%, P ASP were also associated with decreased TG mass (maximal -60%, P ASP profiles in subjects, and suggest that ASP may be regulated through precursor C3 availability, convertase activity and differentiation status. Copyright 2010 Wiley-Liss, Inc.

  8. Tumor budding is a strong and reproducible prognostic marker in T3N0 colorectal cancer.

    LENUS (Irish Health Repository)

    Wang, Lai Mun

    2012-02-01

    BACKGROUND: Tumor budding along the advancing front of colorectal adenocarcinoma is an early event in the metastatic process. A reproducible, prognostic budding scoring system based on outcomes in early stage colorectal cancer has not been established. DESIGN: One hundred twenty-eight T3N0M0 colorectal carcinoma patients with known outcome were identified. Tumor budding was defined as isolated tumor cells or clusters of <5 cells at the invasive tumor front. Tumor bud counts were generated in 5 regions at 200x by 2 pathologists (conventional bud count method). The median bud count per case was used to divide cases into low (median=0) and high budding (median > or =1) groups. Forty cases were reevaluated to assess reproducibility using the conventional and a novel rapid bud count method. RESULTS: Fifty-seven (45%) carcinomas had high and 71 (55%) had low budding scores. High budding was associated with an infiltrative growth pattern (P<0.0001) and lymphovascular invasion (P=0.005). Five-year cancer-specific survival was significantly poorer in high compared with low budding groups: 63% versus 91%, respectively, P<0.0001. Multivariate analysis demonstrated tumor budding to be independently prognostic (hazard ratio=4.76, P<0.001). Interobserver agreement was at least equivalent comparing the conventional to the rapid bud count methods: 87.5% agreement (kappa=0.75) versus 92.5% agreement (kappa=0.85), respectively. CONCLUSIONS: Tumor budding is a strong, reproducible, and independent prognostic marker of outcome that is easily assessed on hematoxylin and eosin slides. This may be useful for identifying the subset of T3N0M0 patients at high risk of recurrence who may benefit from adjuvant therapy.

  9. Comparison of T2 and T3 sympathectomy for compensatory sweating on palmar hyperhidrosis

    Science.gov (United States)

    Turkyilmaz, Atila; Karapolat, Sami; Seyis, Kubra Nur; Tekinbas, Celal

    2017-01-01

    Abstract Background: An otherwise successfully performed endoscopic thoracic sympathectomy (ETS) to treat palmar hyperhidrosis (PH) often has a serious side effect: compensatory sweating (CS). This side effect occurs in other parts of the body to a disturbing extent. The objective of this study is to determine whether there is a relationship between the level of ETS performed on patients with PH, and the occurrence and severity of postoperational CS. Methods: Between January 2014 and January 2015, ETS procedures were performed on 25 randomly selected consecutive subjects (group A) at T2 level, and on another 25 subjects (group B) at T3 level, who all felt severely handicapped due to PH. All subjects were assessed in terms of their demographic characteristics including gender and age, as well as postoperative complications, short-term results, side effects, recurrence of symptoms, and long-term results. Results: The symptoms disappeared in all subjects in short-term, and no recurrence was seen in their short or long-term follow-ups. At the end of year one, CS developed at a rate of 12% in group A and 8% in group B, particularly in their back and abdominal regions. The overall satisfaction with the procedure in year one was 96% in group A and 100% in group B. Conclusion: When an ETS performed at T2 or T3 level for PH involves only the interruption of the sympathetic chain, with a limitation on the range of dissection and avoidance of any damage to ganglia, sweating is stopped completely. No recurrence of PH is encountered, and CS develops only at low rates and severities. PMID:28422886

  10. Overexpression of the mitochondrial T3 receptor induces skeletal muscle atrophy during aging.

    Directory of Open Access Journals (Sweden)

    François Casas

    Full Text Available In previous studies, we characterized a new hormonal pathway involving a mitochondrial T3 receptor (p43 acting as a mitochondrial transcription factor. In in vitro and in vivo studies, we have shown that p43 increases mitochondrial transcription and mitochondrial biogenesis. In addition, p43 overexpression in skeletal muscle stimulates mitochondrial respiration and induces a shift in metabolic and contractile features of muscle fibers which became more oxidative.Here we have studied the influence of p43 overexpression in skeletal muscle of mice during aging. We report that p43 overexpression initially increased mitochondrial mass. However, after the early rise in mitochondrial DNA occurring at 2 months of age in transgenic mice, we observed a progressive decrease of mitochondrial DNA content which became 2-fold lower at 23 months of age relatively to control animals. Moreover, p43 overexpression induced an oxidative stress characterized by a strong increase of lipid peroxidation and protein oxidation in quadriceps muscle, although antioxidant enzyme activities (catalase and superoxide dismutase were stimulated. In addition, muscle atrophy became detectable at 6 months of age, probably through a stimulation of the ubiquitin proteasome pathway via two muscle-specific ubiquitin ligases E3, Atrogin-1/MAFbx and MuRF1.Taken together, these results demonstrate that a prolonged stimulation of mitochondrial activity induces muscle atrophy. In addition, these data underline the importance of a tight control of p43 expression and suggest that a deregulation of the direct T3 mitochondrial pathway could be one of the parameters involved in the occurrence of sarcopenia.

  11. Melatonin Suppresses Autophagy Induced by Clinostat in Preosteoblast MC3T3-E1 Cells.

    Science.gov (United States)

    Yoo, Yeong-Min; Han, Tae-Young; Kim, Han Sung

    2016-04-08

    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.

  12. Resveratrol induces apoptosis and inhibits adipogenesis in 3T3-L1 adipocytes.

    Science.gov (United States)

    Rayalam, Srujana; Yang, Jeong-Yeh; Ambati, Suresh; Della-Fera, Mary Anne; Baile, Clifton A

    2008-10-01

    Resveratrol, a phytoallexin, has recently been reported to slow aging by acting as a sirtuin activator. Resveratrol also has a wide range of pharmacological effects on adipocytes. In this study, we investigated the effects of resveratrol on adipogenesis and apoptosis using 3T3-L1 cells. In mature adipocytes, 100 and 200 microM resveratrol decreased cell viability dose-dependently by 23 +/- 2.7%, and 75.3 +/- 2.8% (p < 0.0001), respectively, after 48 h treatment, and 100 microM resveratrol increased apoptosis by 76 +/- 8.7% (p < 0.0001). Resveratrol at 25 and 50 microM decreased lipid accumulation in maturing preadipocytes significantly by 43 +/- 1.27% and 94.3 +/- 0.3% (p < 0.0001) and decreased cell viability by 25 +/- 1.3% and 70.4 +/- 1.6% (p < 0.0001), respectively. In order to understand the anti-adipogenic effects of resveratrol, maturing 3T3-L1 preadipocytes were treated with 25 microM resveratrol and the change in the expression of several adipogenic transcription factors and enzymes was investigated using real-time RT-PCR. Resveratrol down-regulated the expression of PPAR gamma, C/EBP alpha, SREBP-1c, FAS, HSL, LPL and up-regulated the expression of genes regulating mitochondrial activity (SIRT3, UCP1 and Mfn2). These results indicate that resveratrol may alter fat mass by directly affecting cell viability and adipogenesis in maturing preadipocytes and inducing apoptosis in adipocytes and thus may have applications for the treatment of obesity.

  13. Correlations between age,Charlson score and outcome in clinical unilateral T3a prostate cancer

    Institute of Scientific and Technical Information of China (English)

    Chao-Yu Hsu; Steven Joniau; Raymond Oyen; Tania Roskams; Hein Van Poppel

    2009-01-01

    According to the European Association of Urology (EAU) guidelines,a life expectancy of>10 years is considered an important factor in the treatment of prostate cancer.The Charlson score is used to predict mortality based on comorbidities.The purpose of this study was to investigate the relationship between age,Charlson score and outcome in patients with cT3a prostate cancer.Between 1987 and 2004,200 patients,who were with clinical T3a prostate cancer and who underwent radical prostatectomy (RP),were previously detected by digital rectal examination (DRE).Patients were categorized into two age groups (<65 and≥65 years old).Patients were also divided into two groups according to Charlson score (=0 and≥1).Both age and Charlson score were analyzed regarding their predictive power of patients' outcomes.The mean follow-up period was 70.6 months,and the mean age of patients was 63.3 years.In all,106 patients were<65 years old and 94 patients were≥65 years old.Age was a significant predictor of overall survival (OS).A Charlson score of O was found in 110 patients,and of≥1 in 90 patients.Charlson score was not a significant predictor of biochemical progression-free survival (BPFS),clinical progression-free survival (CPFS) or OS.Cox multivariate analysis showed that margin status was a significant independent factor in BPFS,and cancer volume was a significant independent factor in CPFS.Charlson score does not influence the outcome in patients with clinical locally advanced prostate cancer.Age may influence OS.RP can be performed in motivated healthy older patients.However,the patients need to be counseled regarding possible surgery-related side effects,such as urinary incontinence and erectile dysfunction,which are age-and comorbidity-dependent.

  14. Lack of action of exogenously administered T3 on the fetal rat brain despite expression of the monocarboxylate transporter 8

    OpenAIRE

    Grijota Martínez, María del Carmen; Díez, Diego; Morreale de Escobar, Gabriella; Bernal, Juan; Morte, Beatriz

    2011-01-01

    Mutations of the monocarboxylate transporter 8 gene (MCT8, SLC16A2) cause the Allan-Herndon-Dudley syndrome, an X-linked syndrome of severe intellectual deficit and neurological impairment. Mct8 transports thyroid hormones (T4 and T3), and the Allan-Herndon-Dudley syndrome is likely caused by lack of T3 transport to neurons during critical periods of fetal brain development. To evaluate the role of Mct8 in thyroid hormone action in the fetal brain we administered T4 or T3 to thyroidectomized ...

  15. Clinical significance of BNP,cTnI,T3 combined detection in elderly heart failure%BNP、cTnI、T3联合检测对老年心力衰竭的临床意义

    Institute of Scientific and Technical Information of China (English)

    罗友军; 周新媚; 肖嫒

    2014-01-01

    Objective To investigate the clinical significance of BNP, cTnI, T3 combined detection in elderly heart failure. Methods A total of 138 cases of elderly patients with heart failure (HF group) selected from October 2012 to December 2013 in our hospital were retrospectⅣe analyzed, according to the New York Heart Association (NYHA) classification standards will HF cardiac function of patients 60 cases of class Ⅱ, Ⅲ-Ⅳ grade 78 cases. The control group was 70 cases, were selected to our hospital for physical examination of healthy subjects on the same period with HF patients. BNP, cTnI, T3 were alone tested and joint detected. Results BNP, cTnI levels of NYHA Ⅱ level, NY-HAⅢ-Ⅳ-class before treatment were higher than the control group , T3 levels were lower than the control group;BNP, cTnI levels of NYHAⅢ-Ⅳ-class before treatment were higher than NYHA Ⅱ level, T3 levels were lower than NYHA Ⅱ level; BNP NYHAⅢ-Ⅳ level after treatment, cTnI levels before treatment appeared reduce, T3 levels ap-pear higher, the difference was significant (P<0.05). BNP+cTnI+T3 detect sensitⅣe were higher than BNP, cTnI, T3 alone testing, the difference was significant(P<0.05). Conclusion BNP, cTnI, T3 joint detection has high sensitⅣity for early diagnosis of HF has important significance.%目的:探讨BNP、cTnI、T3联合检测对老年心力衰竭的临床意义。方法回顾性分析2012年10月~2013年12月期间在我院住院治疗的138例老年心力衰竭患者(HF组),按照美国纽约心脏病学会(NYHA)分级标准将HF组患者的心功能分为Ⅱ级60例,Ⅲ~Ⅳ级78例。对照组70例,均为与HF组患者同期来我院进行健康体检的健康研究对象。对患者BNP、cTnI、T3进行单独检测及联合检测。结果 NYHA Ⅱ级、NYHAⅢ~Ⅳ级治疗前的BNP、cTnI水平均高于对照组,T3水平低于对照组;NYHAⅢ~Ⅳ级治疗前的BNP、cTnI水平均高于NYHA Ⅱ级, T3水平低于NYHAⅡ级;NYHA

  16. Development of an 'In vitro' system for the caption essay of T3; Desarrollo de un sistema 'In vitro' para el ensayo de captacion de T3

    Energy Technology Data Exchange (ETDEWEB)

    Lavalley, C.; Ferro, G.; Zambrano, F.; Lezama, J

    1990-02-15

    Triiodothyronine uptake (T3U) is a qualitative technique for evaluation of the unsaturation capacity of thyroid binding globulin (TBG). This paper presents results related to a T3 standardized serum and the integration of T3-I-125, and adsorbent for labelled hormone. Labelled hormone were prepared by the chloramine T method and then purified by high performance liquid chromatography. The specific activity was 500 {mu} Ci/ {mu} g. Various adsorbents such as: Norit A Charcoal, calcium silicate, talc, bovine serum albumin macroaggregated (BSAM) were used in different buffers as: Tris-HCl, barbital and Michaelis. Standardized serum was prepared by mixing different euthyroid sera. Best conditions for T3U assays were achieved with 15 mg/ml. BSAM at pH 8.6 in presence of Tris-HCl buffer for hypothyroid and hyperthyroid sera, for which we obtained < 0.9 {+-} 0.04 and > 1.1 {+-} 0.05 respectively as a T3U index with a 3.0 % of coefficient variation. The reagents so prepared can be conveniently used for T3U assays. (Author)

  17. Archive of Geosample Data and Information from the University of Wisconsin Ice Island T3 Core Collection

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — From 1963 to 1972, 349 piston cores were collected from the Arctic Ocean using Ice Island T3 as a sampling platform and sent to the University of Wisconsin-Madison...

  18. Troponin T3 expression in skeletal and smooth muscle is required for growth and postnatal survival: characterization of Tnnt3(tm2a(KOMP)Wtsi) mice.

    Science.gov (United States)

    Ju, Yawen; Li, Jie; Xie, Chao; Ritchlin, Christopher T; Xing, Lianping; Hilton, Matthew J; Schwarz, Edward M

    2013-09-01

    The troponin complex, which consists of three regulatory proteins (troponin C, troponin I, and troponin T), is known to regulate muscle contraction in skeletal and cardiac muscle, but its role in smooth muscle remains controversial. Troponin T3 (TnnT3) is a fast skeletal muscle troponin believed to be expressed only in skeletal muscle cells. To determine the in vivo function and tissue-specific expression of Tnnt3, we obtained the heterozygous Tnnt3+/flox/lacZ mice from Knockout Mouse Project (KOMP) Repository. Tnnt3(lacZ/+) mice are smaller than their WT littermates throughout development but do not display any gross phenotypes. Tnnt3(lacZ/lacZ) embryos are smaller than heterozygotes and die shortly after birth. Histology revealed hemorrhagic tissue in Tnnt3(lacZ/lacZ) liver and kidney, which was not present in Tnnt3(lacZ/+) or WT, but no other gross tissue abnormalities. X-gal staining for Tnnt3 promoter-driven lacZ transgene expression revealed positive staining in skeletal muscle and diaphragm and smooth muscle cells located in the aorta, bladder, and bronchus. Collectively, these findings suggest that troponins are expressed in smooth muscle and are required for normal growth and breathing for postnatal survival. Moreover, future studies with this mouse model can explore TnnT3 function in adult muscle function using the conditional-inducible gene deletion approach

  19. Influence of Mechanical Force on Bone Matrix Proteins in Ovariectomised Mice and Osteoblast-like MC3T3-E1 Cells

    Science.gov (United States)

    ZHANG, MENG; ISHIKAWA, SHINTARO; INAGAWA, TOMOKO; IKEMOTO, HIDESHI; GUO, SHIYU; SUNAGAWA, MASATAKA; HISAMITSU, TADASHI

    2017-01-01

    Aim: To investigate the effect of mechanical stress on periostin and semaphorin-3A expression in a murine model of postmenopausal osteoporosis and in osteoblast-like MC3T3-E1 cells. Materials and Methods: Female mice were divided into three groups and treated with a sham operation, ovariectomy (OVX) or OVX plus treadmill training (OVX+Run). After 10 weeks, tibias were used for histological analysis. MC3T3-E1 cells were burdened by mechanical stress using a centrifuge or were treated with periostin, and the production of biologically-active semaphorin-3A was examined in vitro. Results: In OVX+Run group tibias, the number of tartrate-resistant acid phosphatase-positive osteoclasts was lower than in the OVX group, and the expression of periostin and semaphorin-3A was higher. In MC3T3-E1 cells, centrifugal stress significantly increased periostin and semaphorin-3A mRNA expression. Treatment with periostin increased the semaphorin-3A level. Conclusion: We speculate that mechanical load may increase periostin production in osteoblasts, and periostin may inhibit osteoclast differentiation by its effects on semaphorin-3A. Our results support the concept of a positive correlation between exercise and inhibition of osteoclasts in post-menopausal osteoporosis. PMID:28064225

  20. The prognostic impact of preoperative blood monocyte count in pathological T3N0M0 rectal cancer without neoadjuvant chemoradiotherapy.

    Science.gov (United States)

    Zhang, Lu-Ning; Xiao, Weiwei; OuYang, Pu-Yun; You, Kaiyun; Zeng, Zhi-Fan; Ding, Pei-Rong; Pan, Zhi-Zhong; Xu, Rui-Hua; Gao, Yuan-Hong

    2015-09-01

    It remains controversial whether adjuvant therapy should be delivered to pathological T3N0M0 rectal cancer without neoadjuvant chemoradiotherapy. Thus identification of patients at high risk is of particular importance. Herein, we aimed to evaluate whether the absolute peripheral blood monocyte count can stratify the pathological T3N0M0M0 rectal cancer patients in survival. A total of 270 pathological T3N0M0 rectal cancer patients with total mesorectal excision-principle radical resection were included. The optimal cut-off value of preoperative monocyte count was determined by receiver operating characteristic curve analysis. Overall survival and disease-free survival between low- and high-monocyte were estimated by Kaplan-Meier method and Cox regression model. The optimal cut-off value for monocyte count was 595 mm(3). In univariate analysis, patients with monocyte counts higher than 595/mm(3) had significantly inferior 5-year overall survival (79.2 vs 94.2 %, P = 0.006) and disease-free survival (67.8 vs 86.0 %, P count remained to be associated with poor overall survival (HR = 2.55, 95 % CI 1.27-5.10; P = 0.008) and disease-free survival (HR = 2.63, 95 % CI 1.48-4.69; P = 0.001). Additionally, the significant association of monocyte count with disease-free survival was hardly influenced in the subgroup analysis, whereas this correlation was restricted to the males and patients with normal carcinoembryonic antigen (CEA) level (count is independently predictive of worse survival of pathological T3N0M0 rectal cancer patients without neoadjuvant chemoradiotherapy. Postoperative adjuvant therapy might be considered for patients with high-monocyte count.

  1. Suppressed intrinsic catalytic activity of GLUT1 glucose transporters in insulin-sensitive 3T3-L1 adipocytes

    Energy Technology Data Exchange (ETDEWEB)

    Harrison, S.A.; Buxton, J.M.; Czech, M.P. (Univ. of Massachusetts Medical Center, Worcester (United States))

    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.

  2. Coculture with BJ fibroblast cells inhibits the adipogenesis and lipogenesis in 3T3-L1 cells

    Energy Technology Data Exchange (ETDEWEB)

    Jeong, Hyun Jeong [Department of Biochemistry, Kwandong University College of Medicine, Gangneung, Gangwondo 210-701 (Korea, Republic of); Park, Sahng Wook [Department of Biochemistry and Molecular Biology, Center for Chronic Metabolic Disease Research, Brain Korea 21 Project for Medical Science, Yonsei University College of Medicine, Seoul 120-752 (Korea, Republic of); Kim, Hojeong [Department of Anatomy, Kwandong University College of Medicine, Gangneung, Gangwondo 210-701 (Korea, Republic of); Park, Sang-Kyu, E-mail: 49park@kd.ac.kr [Department of Biochemistry, Kwandong University College of Medicine, Gangneung, Gangwondo 210-701 (Korea, Republic of); Yoon, Dojun, E-mail: mozart@kd.ac.kr [Department of Biochemistry, Kwandong University College of Medicine, Gangneung, Gangwondo 210-701 (Korea, Republic of)

    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.

  3. Berberine reduces the expression of adipogenic enzymes and inflammatory molecules of 3T3-L1 adipocyte.

    Science.gov (United States)

    Choi, Bong-Hyuk; Ahn, In-Sook; Kim, Yu-Hee; Park, Ji-Won; Lee, So-Young; Hyun, Chang-Kee; Do, Myoung-Sool

    2006-12-31

    Berberine (BBR), an isoquinoline alkaloid, has a wide range of pharmacological effects, yet its exact mechanism is unknown. In order to understand the anti-adipogenic effect of BBR, we studied the change of expression of several adipogenic enzymes of 3T3-L1 cells by BBR treatment. First, we measured the change of leptin and glycerol in the medium of 3T3-L1 cells treated with 1 micrometer, 5 micrometer and 10 micrometer concentrations of BBR. We also measured the changes of adipogenic and lipolytic factors of 3T3-L1. In 3T3-L1 cells, both leptin and adipogenic factors (SREBP-1c, C/EBP-alpha, PPAR-gamma, fatty acid synthase, acetyl-CoA carboxylase, acyl-CoA synthase and lipoprotein lipase) were reduced by BBR treatment. Glycerol secretion was increased, whereas expression of lipolytic enzymes (hormone-sensitive lipase and perilipin) mRNA was slightly decreased. Next, we measured the change of inflammation markers of 3T3-L1 cells by BBR treatment. This resulted in the down-regulation of mRNA level of inflammation markers such as TNF-alpha, IL-6, C- reactive protein and haptoglobin. Taken together, our data shows that BBR has both anti-adipogenic and anti-inflammatory effects on 3T3-L1 adipocytes, and the anti-adipogenic effect seems to be due to the down-regulation of adipogenic enzymes and transcription factors.

  4. NIH 3T3 cells malignantly transformed by mot—2 show inactivation and cytoplasmic sequestration of the p53 protein

    Institute of Scientific and Technical Information of China (English)

    WADHWA; SYUICHITAKANO; 等

    1999-01-01

    In previous studies we have reported that a high level of expression of mot-2 protein results in malignant transformation of NIH 3T3 cells as analyzed by anchorage independent growth and nude mice assays [Kaul et al.,Oncogene,17,907-11,1998].Mot-2 was found to interact with tumor suppressor protein p53.The transient overexpression of mot-2 was inhibitory to transcriptional activation function of p53 [Wadhwa et al.,J.Biol.Chem.,273,2958691,1998].We demonstrate here that mot-2 transfected stable clone of NIH 3T3 that showed malignant properties indeed show inactivation of p53 function as assayed by exogenous p53 dependent reporter.The expression level of p53 in response to UV-irradiation was lower in NIH 3T3/mot-2 as compared to NIH 3T3 cells and also exhibited delay in reachingpeak.Furthermore,upon serum starvation p53 was seen to translocate to the nucleus in NIH 3T3,but not in its mot-3 derivative.The data suggests that mot-2 mediated cytoplasmic sequestration and inactivation of p53 may operate,at least in part,for malignant phenotype of NIH 3T3/mot-2 cells.

  5. Improvement of the BALB/c-3T3 cell transformation assay: a tool for investigating cancer mechanisms and therapies.

    Science.gov (United States)

    Poburski, Doerte; Thierbach, René

    2016-01-01

    The identification of cancer preventive or therapeutic substances as well as carcinogenic risk assessment of chemicals is nowadays mostly dependent on animal studies. In vitro cell transformation assays mimic different stages of the in vivo neoplastic process and represent an excellent alternative to study carcinogenesis and therapeutic options. In the BALB/c-3T3 two-stage transformation assay cells are chemically transformed by treatment with MCA and TPA, along with the final Giemsa staining of morphological aberrant foci. In addition to the standard method we can show, that it is possible to apply other chemicals in parallel to identify potential preventive or therapeutic substances during the transformation process. Furthermore, we successfully combined the BALB/c cell transformation assay with several endpoint applications for protein analysis (immunoblot, subcellular fractionation and immunofluorescence) or energy parameter measurements (glucose and oxygen consumption) to elucidate cancer mechanisms in more detail. In our opinion the BALB/c cell transformation assay proves to be an excellent model to investigate alterations in key proteins or energy parameters during the different stages of transformation as well as therapeutic substances and their mode of action.

  6. Attachment of 3T3 and MDBK cells onto poly(EGDMA/HEMA) based microbeads and their biologically modified forms.

    Science.gov (United States)

    Ayhan, H; Gürhan, I; Pişkin, E

    2000-03-01

    Poly(EGDMA/HEMA) based microbeads were prepared by suspension polymerization. A comonomer, i.e., 2-hydroxyethylmethacrylate (HEMA) was included in the recipe in order to have functional hydroxyl groups on the microbead surfaces. Toluene was used in the polymerization formulations to introduce porosity into the matrix. Hydroxyl groups were first oxidized with NaIO4, and then two biological molecules, namely collagen and fibronectin were immobilized by using glutaraldehyde. A spacer-arm, i.e., hexamethylene diamine, was also used in some cases. More protein molecules were immobilized onto more swellable microbeads using spacer-arm. Higher amounts of collagen were immobilized, more than fibronectin immobilization. Attachment of two cell lines (i.e., 3T3 and MDBK cell lines) on these microbeads with a wide variety of surface properties was studied in vitro culture media. Attachments of both cells even onto the plain microbeads were significant. More cells did attach to more swellable microbeads. Introducing both fibronectin and collagen onto the microbeads caused significant increase in the cell attachment. More cells attached to the microbeads carrying fibronectin covalently attached onto the microbeads through the spacer-arm molecules. Fibronectine was better than collagen for high attachment values. The mathematical model proposed successfully simulated attachment kinetics.

  7. Importância do co-cultivo com fibroblastos de camundongo 3T3 para estabelecer cultura de suspensão de células epiteliais do limbo humano Importance of 3T3 feeder layer to establish epithelial cultures from cell suspension obtained from corneo-scleral rims

    Directory of Open Access Journals (Sweden)

    Priscila Cardoso Cristovam

    2008-10-01

    segments was placed with the epithelial side up on the bottom of a 6-well culture plate (Group A. The other two fragments were trypsinized and the obtained cell suspension was cultured with (Group B or without (Group C irradiaded 3T3 cells. The cells were cultured in supplemental hormonal epithelial medium (SHEM, the epithelial migration and clone formation in groups A, B and C were evaluated with phase contrast microscopy and rodamine B staining. RESULTS: Epithelial cell growth was observed in 4/6 rims (Group A. All epithelial cell suspensions that were cultured with 3T3 cells (Group B formed clones. No adhesion or true clone formation (holo- or meroclones was observed in the cell suspensions that were cultivated without 3T3 (Group C (p=0.009. CONCLUSIONS: Epithelial cell suspension obtained from corneo-scleral rims in this model needs to be cultivated with 3T3 cells in order to form clones and establish limbal epithelial cell colonies with the potential to be used for ocular surface reconstruction.

  8. mVps45 knockdown selectively modulates VAMP expression in 3T3-L1 adipocytes.

    Science.gov (United States)

    Sadler, Jessica B A; Roccisana, Jennifer; Virolainen, Minttu; Bryant, Nia J; Gould, Gwyn W

    2015-01-01

    Insulin stimulates the delivery of glucose transporter-4 (GLUT4)-containing vesicles to the surface of adipocytes. Depletion of the Sec1/Munc18 protein mVps45 significantly abrogates insulin-stimulated glucose transport and GLUT4 translocation. Here we show that depletion of mVps45 selectively reduced expression of VAMPs 2 and 4, but not other VAMP isoforms. Although we did not observe direct interaction of mVps45 with any VAMP isoform; we found that the cognate binding partner of mVps45, Syntaxin 16 associates with VAMPs 2, 4, 7 and 8 in vitro. Co-immunoprecipitation experiments in 3T3-L1 adipocytes revealed an interaction between Syntaxin 16 and only VAMP4. We suggest GLUT4 trafficking is controlled by the coordinated expression of mVps45/Syntaxin 16/VAMP4, and that depletion of mVps45 regulates VAMP2 levels indirectly, perhaps via reduced trafficking into specialized subcellular compartments.

  9. Hsp90 chaperones PPARγ and regulates differentiation and survival of 3T3-L1 adipocytes.

    Science.gov (United States)

    Nguyen, M T; Csermely, P; Sőti, C

    2013-12-01

    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.

  10. ENHANCEMENT OF NIH3T3 CELL PROLIFERATION BY EXPRESSING MACROPHAGE COLONY STIMULATING FACTOR IN NUCLEI

    Institute of Scientific and Technical Information of China (English)

    曹震宇; 吴克复; 李戈; 林永敏; 张斌; 郑国光

    2003-01-01

    Objective: To explore the effects of nuclear M-CSF on the process of tumorigenesis. Methods: Functional part of M-CSF cDNA was inserted into an eukaryotic expression plasmid pCMV/myc/nuc, which can add three NLS to the C-terminal of the expressed protein and direct the protein into the cell nuclei. The constructed plasmid was transferred into NIH3T3 cells and the cell clones were selected by G-418 selection. Cell clones stable expressing target protein were identified by RT-PCR, ABC immunohistochemistry assay and Western blot. Cell growth kinetics analyses through growth curves, cell doubling time, MTT test and anti-sense oligodeoxynucleotide (ASODN) inhibiting cell growth test were performed to identify cells proliferation potential. Results: The transfected cells showed elevated proliferation potential over the control cells. Conclusion: Abnormal appearance of M-CSF in nucleus could enhance cell proliferation, which suggests that cytokine isoforms within cell nucleus might play transcription factor-like role.

  11. Corrosion inhibition mechanisms of aluminum alloy 2024-T3 by selected non-chromate inhibitors

    Science.gov (United States)

    Lopez-Garrity, Omar A.

    The pursuit to find a chromate-alternative has led to the development of several chromate-free aerospace primers and coating systems that offer good protection. However, fundamental understanding of the functionality of the chromate-free pigments that are embedded within these coating systems is lacking. The objective of this study was to understand the fundamental mechanism of corrosion inhibition of aluminum alloy 2024-T3 by molybdate (MoO 42-), silicate (SiO32-), and praseodymium (Pr3+) with the goal of developing the kind of understanding that was accomplished for chromate. Furthermore, since most inhibiting conversion coatings and pigments act by releasing soluble species into the local environment, it was of interest to understand the mechanism of inhibition in aqueous 0.1 M NaCl solution. The mechanism of inhibition of AA2024-T3 by the select non-chromate inhibitors was investigated using various electrochemical, microscopic and spectroscopic techniques. Naturally aerated polarization curves showed that molybdate provided mixed inhibition in near-neutral pH and at a threshold concentration of 0.1 M. The largest effect was a 250 mV increase in the breakdown potential associated with pitting and a 350 mV decrease in the open-circuit potential (OCP). In addition, electrochemical impedance indicated that the corrosion inhibition mechanism is oxygen-dependent owing to the protection afforded by Mo(VI) species. It was proposed that the corrosion inhibition of AA2024-T3 by molybdate may occur following a two-step process whereby molybdate is rapidly reduced to MoO.(OH)2 over the intermetallic particles and is subsequently oxidized to intermediate molybdenum oxides (e.g. Mo4O11) in the presence of oxygen which is reduced. This in turn may lead to a local acidification, promoting the condensation and polymerization of molybdate species in solution to form polymolybdate species (Mo7O24 6- and Mo8O264-). Furthermore, S-phase particle dissolution is decreased

  12. Adhesion, Proliferation and Migration of NIH/3T3 Cells on Modified Polyaniline Surfaces

    Science.gov (United States)

    Rejmontová, Petra; Capáková, Zdenka; Mikušová, Nikola; Maráková, Nela; Kašpárková, Věra; Lehocký, Marián; Humpolíček, Petr

    2016-01-01

    Polyaniline shows great potential and promises wide application in the biomedical field thanks to its intrinsic conductivity and material properties, which closely resemble natural tissues. Surface properties are crucial, as these predetermine any interaction with biological fluids, proteins and cells. An advantage of polyaniline is the simple modification of its surface, e.g., by using various dopant acids. An investigation was made into the adhesion, proliferation and migration of mouse embryonic fibroblasts on pristine polyaniline films and films doped with sulfamic and phosphotungstic acids. In addition, polyaniline films supplemented with poly (2-acrylamido-2-methyl-1-propanesulfonic) acid at various ratios were tested. Results showed that the NIH/3T3 cell line was able to adhere, proliferate and migrate on the pristine polyaniline films as well as those films doped with sulfamic and phosphotungstic acids; thus, utilization of said forms in biomedicine appears promising. Nevertheless, incorporating poly (2-acrylamido-2-methyl-1-propanesulfonic) acid altered the surface properties of the polyaniline films and significantly affected cell behavior. In order to reveal the crucial factor influencing the surface/cell interaction, cell behavior is discussed in the context of the surface energy of individual samples. It was clearly demonstrated that the lesser the difference between the surface energy of the sample and cell, the more cyto-compatible the surface is. PMID:27649159

  13. Investigation on a Sol-gel Coating Containing Inhibitors on 2024-T3 Aluminum Alloy

    Institute of Scientific and Technical Information of China (English)

    SHI Hong-wei; LIU Fu-chun; HAN En-hou; SUN Ming-cheng

    2006-01-01

    For a long time, chromate incorporated conversion coatings have been drawn special attention in corrosion protection of aircraft-used aluminum alloys. However,ever-increasing environmental pressures requires that non-chromate conversion coatings be developed because of the detrimental carcinogenic effects of the chromate compounds. In recent years, the sol-gel coatings doped with inhibitors were developed to replace chromate conversion coatings, and showed real promise. A sol-gel coating was prepared and its anti-corrosion behavior was investigated using the potentiodynamic scanning (PDS) and the electrochemical impedance spectroscopy (EIS). It is found that the sol-gel coating obtained by the hydrolysis and condensation of 3-glycidoxypropyltrimethoxysilane (GPTMS) and tetramethoxysilane (TMOS) is prone to form defects if cured at the room temperature, whereas if cured at a higher temperature (100 ℃), these flaws can be avoided. Furthermore, it can be seen that addition of anti-foam agents and surfactants will reduce the faults if cured at the room temperature. Effects of the corrosion inhibitors, CeCl3 and mercaptobenzothiazole (MBT), in the sol-gel coatings on 2024-T3 aluminum alloy were also investigated. Results show that the corrosion resistance of the sol-gel coatings containing CeCl3 proves to be better than that of the pure and MBT added sol-gel coatings by the electrochemical methods.

  14. Intracytoplasmic triglyceride accumulation produced by dexamethasone in adult rat hepatocytes cultivated on 3T3 cells.

    Science.gov (United States)

    Mendoza-Figueroa, T; Hernandez, A; De Lourdes Lopez, M; Kuri-Harcuch, W

    1988-11-30

    Glucocorticoids, such as hydrocortisone (HC) and dexamethasone (DEX), when administered to rats, induce lipid accumulation within hepatocytes (fatty liver). To investigate whether glucocorticoids can produce triglyceride (TG) accumulation as they do in vivo and the involved mechanisms, we have used primary cultures of rat hepatocytes which synthesized and secrete triglycerides into the culture medium. Hepatocytes cultivated on a feeder layer of lethally treated 3T3 cells were exposed for 2 weeks to micromolar concentrations of DEX. This glucocorticoid caused morphological alterations and cells accumulated lipid droplets in their cytoplasm; the TG content increased up to 6-fold in a concentration- and time-dependent manner. The removal of [14C]acetic or [14C]oleic acid from the culture medium was not altered in the cultures treated with 50 micrograms/ml DEX but the incorporation of [14C]acetic and [14C]oleic acid into TG in these cultures was about 13-fold and 60% higher than in non-treated cells, respectively. On the other hand, hepatocytes treated with 50 micrograms/ml DEX for 2 weeks showed a 16-fold decrease in TG release and 40% inhibition in protein export, whereas synthesis of total cellular proteins was not altered. Our results show that corticosteroids, such as DEX, caused lipid accumulation in liver cells through an increased synthesis and/or esterification of fatty acids, but mostly through a decrease in the secretion of TG.

  15. Cultivation of adult rat hepatocytes on 3T3 cells: expression of various liver differentiated functions.

    Science.gov (United States)

    Kuri-Harcuch, W; Mendoza-Figueroa, T

    1989-08-01

    Adult rat hepatocytes were maintained in culture for at least 1 month without losing the expression of their differentiated functions; they were cultured on lethally treated 3T3 fibroblasts inoculated at 35,000 cells/cm2 with medium containing 10-25 micrograms/ml hydrocortisone. Hepatocytes showed their typical morphology; they formed bile canaliculi, microvilli, and intercellular junctions with desmosomes and nexus; some formed structures that may resemble the perisinusoidal space of Disse. In addition, they showed DNA synthesis and expressed some liver-specific functions. They synthesized albumin and other proteins, which were exported to the culture medium. Like parenchymal liver cells in vivo, de novo fatty acid synthesis and esterification took place, and more than 80% of the lipids synthesized by the hepatocytes were secreted into the medium as triglycerides; they also showed cytochrome-P450 activity that was inducible with phenobarbital, suggesting that the hepatocytes have the capacity to metabolize drugs. These culture conditions allow the study of various hepatocyte differentiated functions, and they may provide the means to analyze the effect on liver of hormones, viruses and hepatotoxic chemicals and drugs; they may also indicate conditions adequate for serial growth of hepatocytes.

  16. Culture of proliferating and differentiating fat-storing cells in 3T3-conditioned medium.

    Science.gov (United States)

    Mendoza-Figueroa, T; Argüello, C; Kuri-Harcuch, W

    1988-01-01

    There is growing evidence suggesting that hepatic fat-storing cells (FSC) or Ito cells have an important function in vitamin A storage and metabolism and in the synthesis of connective tissue components in normal liver and during fibrogenesis. The purified FSC acquire a fibroblastic morphology and their vitamin A content decreases in culture. We cultivated cells under in vitro conditions that allowed the expression of FSC morphological and functional characteristics for 3-4 weeks of primary culture. Cells were isolated from rat liver by the collagenase-perfusion method without further purification and cultured with 3T3-conditioned medium, which seemed to stimulate the selective proliferation of the FSC. After 8-10 days, round and stellate cells grew actively from a few precursor cells in the primary culture and were not subcultivated; the stellate cells had the ability to become round and vice versa and were highly motile. The cells had intracytoplasmic lipid droplets, a well developed rough endoplasmic reticulum, Golgi complex, numerous vesicles filled with electron-dense material, and extracellular matrix (ECM) components on their surface. Both stellate and round cells showed the presence of desmin by immunofluorescence and vitamin A autofluorescence, but lacked peroxidase activity. The culture conditions we describe allowed the selective proliferation of cells with morphological and functional characteristics of the FSC in the normal liver, raising the possibility of studying FSC proliferation and differentiation.

  17. Evaluating protection systems against marine corrosion of aeronautic alloy Alclad 2024-T3

    Directory of Open Access Journals (Sweden)

    William Aperador Chaparro

    2011-05-01

    Full Text Available  This paper shows how two coating systems were obtained as an alternative for protection against corrosion of al clad 2024-T3which is used in battery compartment manufacture for T-41 aircraft. Such systems consist of three types of organic resin: a first layer of P-115 polyester resin as the first coating on both systems, and a second layer of Hetron 197-3 polyester resin in the first system and vinyl-ester resin in the second one. Scanning electron microscopy (SEM was used for surface morphology analysis, showing the roughness produced by surface treatment. The coatings were electrochemically characterised by electro chemical impedance spectroscopy (EIS and Tafel polarization curves; it was found that both systems had good performance against corrosion in a marine environment and the chemical surface preparation system had a superior protective pattern for Alodine5700 + 197-3 Hetron, a 1.42x10-12mpycorrosion rate being obtained while substratum rate was 1.59x10-7 mpy. 

  18. Echinacea purpurea root extract enhances the adipocyte differentiation of 3T3-L1 cells.

    Science.gov (United States)

    Shin, Dong-Mi; Choi, Kyeong-Mi; Lee, Youn-Sun; Kim, Wonkyun; Shin, Kyong-Oh; Oh, Seikwan; Jung, Jae-Chul; Lee, Mi Kyeong; Lee, Yong-Moon; Hong, Jin Tae; Yun, Yeo-Pyo; Yoo, Hwan-Soo

    2014-06-01

    Echinacea purpurea has been shown to have anti-diabetic activities; for example, it activates peroxisome proliferator-activated receptor γ (PPARγ) and increases insulin-stimulated glucose uptake. Adipogenesis has been used to study the insulin signaling pathway and to screen anti-diabetic compounds. The present study was conducted to investigate the effects of an ethanol extract of E. purpurea (EEEP) and its constituents on the insulin-induced adipocyte differentiation of 3T3-L1 preadipocytes. When adipocyte differentiation was induced with insulin plus 3-isobutyl-1-methylxanthine and dexamethasone, the accumulation of lipid droplets and the cellular triglyceride content were significantly increased by EEEP. The expressions of PPARγ and C/EBPα in adipocytes treated with EEEP were gradually increased as compared with control cells. Fat accumulation and triglyceride content of adipocytes treated with dodeca-2(E),4(E)-dienoic acid isobutylamide were significantly increased as compared with control cells. The expressions of PPARγ and C/EBPα in adipocytes treated with dodeca-2(E),4(E)-dienoic acid isobutylamide were significantly higher than in control cells. These results suggest EEEP promotes the adipogenesis that is partially induced by insulin and that dodeca-2(E),4(E)-dienoic acid isobutylamide appears to be responsible for EEEP-enhanced adipocyte differentiation.

  19. Effect of CrO3 Sealing Time on Anodized A12024-T3

    Science.gov (United States)

    Korda, Akhmad A.; Hidayat, R. Z.

    2016-08-01

    The effect of CrO3 sealing time on anodized aluminum alloy has been investigated. A1 2024-T3 were used as substrate. Anodizing was carried out using chromic acid. CrO3 sealing was conducted in CrO3 solution for 30, 60, 90, 120 and 150 minutes. As comparison, other specimens were also prepared as anodized and boiled water sealing. Thickness of the coating was observed by optical microscope. Anodized and sealing layer was analyzed by X- ray diffraction. The hardness of as anodized, boiled water sealing and CrO3 sealing were compared. The highest hardness is achieved by CrO3 sealed specimen and followed by boiled water sealing and as anodized specimens. The longer the processes of CrO3 sealing the higher layer thickness and therefore the higher hardness of the oxide layer. The best resistance to electrolyte penetration is achieved by the CrO3 sealed specimen followed by boiled water sealed and as anodized specimens. The higher thickness of oxide layer, the higher the resistance against electrolyte penetration.

  20. Hierarchical polymeric scaffolds support the growth of MC3T3-E1 cells.

    Science.gov (United States)

    Akbarzadeh, Rosa; Minton, Joshua A; Janney, Cara S; Smith, Tyler A; James, Paul F; Yousefi, Azizeh-Mitra

    2015-02-01

    Tissue engineering makes use of the principles of biology and engineering to sustain 3D cell growth and promote tissue repair and/or regeneration. In this study, macro/microporous scaffold architectures have been developed using a hybrid solid freeform fabrication/thermally induced phase separation (TIPS) technique. Poly(lactic-co-glycolic acid) (PLGA) dissolved in 1,4-dioxane was used to generate a microporous matrix by the TIPS method. The 3D-bioplotting technique was used to fabricate 3D macroporous constructs made of polyethylene glycol (PEG). Embedding the PEG constructs inside the PLGA solution prior to the TIPS process and subsequent extraction of PEG following solvent removal (1,4-dioaxane) resulted in a macro/microporous structure. These hierarchical scaffolds with a bimodal pore size distribution (300 μm) contained orthogonally interconnected macro-channels generated by the extracted PEG. The diameter of the macro-channels was varied by tuning the dispensing parameters of the 3D bioplotter. The in vitro cell culture using murine MC3T3-E1 cell line for 21 days demonstrated that these scaffolds could provide a favorable environment to support cell adhesion and growth.

  1. Modulation of Osteogenesis in MC3T3-E1 Cells by Different Frequency Electrical Stimulation.

    Directory of Open Access Journals (Sweden)

    Yu Wang

    Full Text Available 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.

  2. Influence of friction stir welding parameters on properties of 2024 T3 aluminium alloy joints

    Directory of Open Access Journals (Sweden)

    Eramah Abdsalam M.

    2014-01-01

    Full Text Available The aim of this work is to analyse the process of friction stir welding (FSW of 3mm thick aluminium plates made of high strength aluminium alloy - 2024 T3, as well as to assess the mechanical properties of the produced joints. FSW is a modern procedure which enables joining of similar and dissimilar materials in the solid state, by the combined action of heat and mechanical work. This paper presents an analysis of the experimental results obtained by testing the butt welded joints. Tensile strength of the produced joints is assessed, as well as the distribution of hardness, micro-and macrostructure through the joints (in the base material, nugget, heat affected zone and thermo-mechanically affected zone. Different combinations of the tool rotation speed and the welding speed are used, and the dependence of the properties of the joints on these parameters of welding technology is determined. [Projekat Ministarstva nauke Republike Srbije, br. TR 34018 i br. TR 35006

  3. Fisetin induces Sirt1 expression while inhibiting early adipogenesis in 3T3-L1 cells.

    Science.gov (United States)

    Kim, Sang Chon; Kim, Yoo Hoon; Son, Sung Wook; Moon, Eun-Yi; Pyo, Suhkneung; Um, Sung Hee

    2015-11-27

    Fisetin (3,7,3',4'-tetrahydroxyflavone) is a naturally found flavonol in many fruits and vegetables and is known to have anti-aging, anti-cancer and anti-viral effects. However, the effects of fisetin on early adipocyte differentiation and the epigenetic regulator controlling adipogenic transcription factors remain unclear. Here, we show that fisetin inhibits lipid accumulation and suppresses the expression of PPARγ in 3T3-L1 cells. Fisetin suppressed early stages of preadipocyte differentiation, and induced expression of Sirt1. Depletion of Sirt1 abolished the inhibitory effects of fisetin on intracellular lipid accumulation and on PPARγ expression. Mechanistically, fisetin facilitated Sirt1-mediated deacetylation of PPARγ and FoxO1, and enhanced the association of Sirt1 with the PPARγ promoter, leading to suppression of PPARγ transcriptional activity, thereby repressing adipogenesis. Lowering Sirt1 levels reversed the effects of fisetin on deacetylation of PPARγ and increased PPARγ transactivation. Collectively, our results suggest the effects of fisetin in increasing Sirt1 expression and in epigenetic control of early adipogenesis. Copyright © 2015 Elsevier Inc. All rights reserved.

  4. Withaferin A induces apoptosis and inhibits adipogenesis in 3T3-L1 adipocytes.

    Science.gov (United States)

    Park, Hea Jin; Rayalam, Srujana; Della-Fera, Mary Anne; Ambati, Suresh; Yang, Jeong-Yeh; Baile, Clifton A

    2008-01-01

    Withaferin A (WA), a highly oxygenated steroidal lactone that is found in the medicinal plant Withania somnifera (also called ashwagandha) has been reported to have anti-tumor, anti-angiogenesis, and pro-apoptotic activity. We investigated the effects of WA on viability, apoptosis and adipogenesis in 3T3-L1 adipocytes. Pre- and post-confluent preadipocytes and mature adipocytes were treated with WA (1-25 microM) up to 24 hrs. Viability and apoptosis were measured by CellTiter-Blue Cell Viability Assay and single strand DNA ELISA Assay, respectively. WA decreased viability and induced apoptosis in all stages of cells. Induction of apoptosis by WA in mature adipocytes was mediated by increased ERK1/2 phosphorylation and altered Bax and Bcl2 protein expression. The effect of WA on adipogenesis was examined by AdipoRed Assay after treating with WA (0.1-1 microM) during the differentiation period. WA decreased lipid accumulation in a dose-dependent manner and decreased the expression of peroxisome proliferator-activated receptor gamma, CCAAT/enhancer binding protein alpha and adipocyte fatty acid binding protein. The effects on apoptosis and lipid accumulation were also confirmed with Hoechst staining and Oil Red O staining, respectively. These results show that WA acts on adipocytes to reduce cell viability and adipogenesis and also induce apoptosis.

  5. Effects of leptin on apoptosis and adipogenesis in 3T3-L1 adipocytes.

    Science.gov (United States)

    Ambati, Suresh; Kim, Hye-Kyeong; Yang, Jeong-Yeh; Lin, Ji; Della-Fera, Mary Anne; Baile, Clifton A

    2007-02-01

    Leptin has been demonstrated to induce adipose tissue apoptosis, which can contribute to the decrease of adiposity, after either central nervous system or peripheral administration. However, it is not known whether leptin acts only centrally to initiate a signal or can also act directly on adipocytes to induce apoptosis. The objective of this study was to determine the direct effect of leptin on adipocyte apoptosis and adipogenesis in vitro using 3T3-L1 cell lines. An ELISA for single stranded DNA, which is highly specific for apoptotic cells, was used to quantify apoptosis. Preconfluent preadipocytes treated with 10(-9), 10(-8), 10(-7), and 10(-6)M leptin showed inhibitory effects on cell viability, and similar observations were also found in maturing preadipocytes treated during day 0-2 and day 2-4 of maturation. After 48 h incubation with 10(-6)M leptin, LDH release was increased by 24.3% (p<0.05) in preconfluent preadipocytes and by 108.5% (p<0.01) in maturing preadipocytes. However, ssDNA analysis revealed no increased apoptosis in preconfluent or maturing preadipocytes or in mature adipocytes treated with leptin. Leptin significantly reduced lipid accumulation and GPDH activity in maturing preadipocytes, demonstrating an inhibitory effect of leptin on adipogenesis. These results indicate that leptin does not act directly to induce adipocyte apoptosis, but can act directly to inhibit maturation of preadipocytes.

  6. Anti-obesity effects of xanthohumol plus guggulsterone in 3T3-L1 adipocytes.

    Science.gov (United States)

    Rayalam, Srujana; Yang, Jeong-Yeh; Della-Fera, Mary Anne; Park, Hea Jin; Ambati, Suresh; Baile, Clifton A

    2009-08-01

    Xanthohumol (XN) and guggulsterone (GS) have each been shown to inhibit adipogenesis and induce apoptosis in adipocytes. In the present study effects of the combination of XN + GS on 3T3-L1 adipocyte apoptosis and adipogenesis were investigated. Mature adipocytes were treated with XN and GS individually and in combination. XN and GS individually decreased cell viability, but XN + GS caused an enhanced decrease in viability and potentiated induction of apoptosis. Likewise, XN + GS caused a potentiated increase in caspase-3/7 activation, whereas neither of the compounds showed any effect individually. In addition, western blot analysis revealed that XN + GS increased Bax expression and decreased Bcl-2 expression, whereas individual compounds did not show any significant effect. XN and GS both decreased lipid accumulation. Individually, XN at 1.5 microM and GS at 3.12 microM decreased lipid accumulation by 26 +/- 4.5% (P < .001) each, whereas XN1.5 + GS3.12 decreased lipid accumulation by 78.2 +/- 1.8% (P < .001). Moreover, expression of the adipocyte-specific proteins was down-regulated with XN1.5 + GS3.12, but no effect was observed with the individual compounds. Finally, XN + GS caused an enhanced stimulation of lipolysis. Thus, combination of XN and GS is more potent in exerting anti-obesity effects than additive effects of the individual compounds.

  7. MC3T3-E1 Cells on Titanium Surfaces with Nanometer Smoothness and Fibronectin Immobilization

    Directory of Open Access Journals (Sweden)

    Tohru Hayakawa

    2012-01-01

    Full Text Available The present study was aimed to evaluate the viability and total protein contents of osteoblast-like cells on the titanium surface with different surface mechanical treatment, namely, nanometer smoothing (Ra: approximately 2.0 nm and sandblasting (Ra: approximately 1.0 μm, and biochemical treatment, namely, with or without fibronectin immobilization. Fibronectin could be easily immobilized by tresyl chloride-activation technique. MC3T3-E1 cells were seeded on the different titanium surfaces. Cell viability was determined by MTT assay. At 1 day of cell culture, there were no significant differences in cell viability among four different titanium surfaces. At 11 days, sandblasted titanium surface with fibronectin immobilization showed the significantly highest cell viability than other titanium surface. No significant differences existed for total protein contents among four different titanium surfaces at 11 days of cell culture. Scanning electron microscopy observation revealed that smoothness of titanium surface produced more spread cell morphologies, but that fibronectin immobilization did not cause any changes of the morphologies of attached cells. Fibronectin immobilization provided greater amount of the number of attached cells and better arrangement of attached cells. In conclusion, the combination of sandblasting and fibronectin immobilization enhanced the cell viability and fibronectin immobilization providing better arrangements of attached cells.

  8. The role of Akt on Arsenic trioxide suppression of 3T3-L1 preadipocyte differentiation

    Institute of Scientific and Technical Information of China (English)

    Zhi Xin WANG; Chun Sun JIANG; Lei LIU; Xiao Hui WANG; Hai Jing JIN; Qiao WU; Quan CHEN

    2005-01-01

    The present study investigates the molecular details of how arsenic trioxide inhibits preadipocyte differentiation and examines the role of Akt/PKB in regulation of differentiation and apoptosis. Continual exposure of arsenic trioxide, at the clinic achievable dosage that does not induce apoptosis, suppressed 3T3-L1 cell differentiation into fat cells by inhibiting the expression of PPARγ and C/EBPα and disrupting the interaction between PPARγ and RXRα, which determines the programming of the adipogenic genes. Interestingly, if we treated the cells for 12 or 24 h and then withdrew arsenic trioxide, the cells were able to differentiate to the comparable levels of untreated cells as assayed by the activity of GAPDH, the biochemical marker of preadipocyte differentiation. Long term treatment blocked the differentiation and the activity of GAPDH could not recover to the comparable levels of untreated cells. Continual exposure of arsenic trioxide caused accumulation in G2/M phase and the accumulation of p21. We found that arsenic trioxide induced the expression and the phosphorylation of Akt/PKB and it inhibited the interaction between Akt/PKB and PPARγ. Akt/PKB inhibitor appears to block the arsenic trioxide suppression of differentiation. Our results suggested that Akt/PKB may play a role in suppression of apoptosis and negatively regulate preadipocyte differentiation.

  9. [Thyroxine (T4) and tri-iodothyronine (T3) determinations: techniques and value in the assessment of thyroid function].

    Science.gov (United States)

    Sapin, R; Schlienger, J-L

    2003-01-01

    Hormonal production of the thyroid gland is constituted of thyroxine or T4 (80%) and triiodothyronine or T3 (20%). In the circulation, whole T4 originates from thyroid secretion but most of T3 (80%) is produced extrathyroidally from T4 deiodination. Conversion of T4 to T3 may be influenced by various conditions and circulating T3 is a less reliable reflection of thyroid hormone production than T4. In serum most of T4 and T3 is bound to binding proteins and only 0.02% of T4 and 0.3% of T3 is free. Because of their higher diagnostic performance, free T4 (FT4) and free T3 (FT3) measurements have superseded total (free + bound) hormone determination. Total hormone measurements remain useful for research studies or in case of severe hyperthyroidism. Equilibrium dialysis/RIA is considered as the reference method for free hormone measurements. Routine clinical laboratories use automated direct two-step or one-step immunoassays with a high molecular weight ligand or labelled antibody. Free hormone measurement remains technically demanding, especially in sera from severe non-thyroid ill patients with low serum thyroxine binding capacity. Interference from anti-thyroid hormone antibodies and familial dysalbuminemic hyperthyroxinemia depends on the assay method, but is now less marked and less frequently detected. To be able to correctly interpret the results of an assay, it is necessary to assess its performance in biologically and clinically well-characterised serum samples. FT4, and FT3 measurements, if FT4 is normal and hyperthyroidism suspected, are used to confirm and assess the level of hypo and hyperthyroidism (overt or subclinical). When the thyroidal status is unstable (first months of a thyroid treatment, altered L-T4 dose, subacute thyroiditis) or when the hypothalamic-pituitary function is disturbed (central hypothyroidism), TSH determination is diagnostically misleading and only free hormone measurements are reliable for thyroid function assessment.

  10. Blueberry peel extracts inhibit adipogenesis in 3T3-L1 cells and reduce high-fat diet-induced obesity.

    Directory of Open Access Journals (Sweden)

    Yuno Song

    Full Text Available This study examined the anti-obesity effect and mechanism of action of blueberry peel extracts (BPE in 3T3-L1 cells and high-fat diet (HFD-induced obese rats. The levels of lipid accumulation were measured, along with the changes in the expression of genes and proteins associated with adipocyte differentiation in 3T3-L1 cells. Evidenced by Oil-red O staining and triglyceride assay, BPE dose-dependently inhibited lipid accumulation at concentrations of 0, 50, and 200 µg/ml. BPE decreased the expression of the key adipocyte differentiation regulator C/EBPβ, as well as the C/EBPα and PPARγ genes, during the differentiation of preadipocytes into adipocytes. Moreover, BPE down-regulated adipocyte-specific genes such as aP2 and FAS compared with control adipocytes. The specific mechanism mediating the effects of BP revealed that insulin-stimulated phosphorylation of Akt was strongly decreased, and its downstream substrate, phospho-GSK3β, was downregulated by BPE treatment in 3T3-L1 cells. Together, these data indicated that BP exerted anti-adipogenic activity by inhibiting the expression of PPARγ and C/EBPβ and the Akt signaling pathway in 3T3-L1 adipocytes. Next, we investigated whether BP extracts attenuated HFD-induced obesity in rats. Oral administration of BPE reduced HFD-induced body weight gain significantly without affecting food intake. The epididymal or perirenal adipose tissue weights were lower in rats on an HFD plus BPE compared with the tissue weights of HFD-induced obese rats. Total cholesterol and triglyceride levels in the rats fed BPE were modestly reduced, and the HDL-cholesterol level was significantly increased in HFD plus BP-fed rats compared with those of HFD-fed rats. Taken together, these results demonstrated an inhibitory effect of BP on adipogenesis through the down-regulation of C/EBPβ, C/EBPα, and PPARγ and the reduction of the phospho-Akt adipogenic factor in 3T3-L1 cells. Moreover, BPE reduced body weight gain

  11. Tmed2 accelerates murine MC3T3-E1 pre-osteoblast proliferation in vitro%Tmed2促进体外小鼠前成骨细胞MC3T3-E1增殖

    Institute of Scientific and Technical Information of China (English)

    熊元; 熊霞辉; 卢雅彬; 朱宁; 陈梅红

    2012-01-01

    目的 研究Tmed2基因对小鼠前成骨细胞增殖的影响.方法1)分别在小鼠MC3T3-E1细胞中过表达和抑制Tmed2,检测细胞增殖情况.2)用雌激素处理细胞后,检测细胞的增殖及Tmed2基因的表达量.荧光实时定量PCR检测mRNA水平,MTS法检测细胞活力和增殖,流式细胞术检测细胞周期,Western blot法检测蛋白水平.结果 过表达Tmed2使MC3T3 -E1细胞的增殖速度加快,细胞周期中S期细胞比例明显增加,且Cyclin A的表达升高.而抑制Tmed2基因表达使MC3T3-E1细胞的增殖速度减慢.雌激素处理使细胞增殖速度加快的同时,Tmed2基因的表达显著增高.结论 Tmed2通过上调Cyclin A的表达,使S期细胞比例增加,加快小鼠前成骨细胞MC3T3-E1的增殖.此外,Tmed2的表达受雌激素的调控,可能参与雌激素促进MC3T3-E1细胞增殖的作用.%Objective To study the role of Tmed2 in murine pre-osteoblast proliferation. Methods 1) Over-express and knock down Tmed2, and the proliferation rate of MC3T3-E1 cell was then detected. 2) MC3T3-E1 cells were treated with p-Estradiol, cell number was then counted and the mRNA level of Tmed2 was analyzed. Quantitative real-time PCR was used to measure the mRNA level, MTS assay was used to assess the cell proliferation rate, flow eytometry was used to figure out the cell cycle distribution, and the protein level was evaluated by Western blot. Results Over-expression of Tmed2 accelerated MC3T3-E1 cell proliferation, and the proportion of cells in S phase markedly increased. The expression of Cyclin A also increased. On the contrary, Knockdown of Tmed2 decelerated MC3T3-E1 cell proliferation. In β-Estradiol promoted MC3T3-E1 proliferation, the mRNA level of Tmed2 remarkably increased. Conclusions Tmed2 accelerates murine pre-osteoblast MC3T3-E1 proliferation through up-regulating Cyclin A expression and therefore increasing the proportion of cells in S phase. In addition, the expression of Tmed2 is regulated by

  12. Corrosion inhibition by inorganic cationic inhibitors on the high strength alumunium alloy, 2024-T3

    Science.gov (United States)

    Chilukuri, Anusha

    The toxicity and carcinogenic nature of chromates has led to the investigation of environmentally friendly compounds that offer good corrosion resistance to AA 2024-T3. Among the candidate inhibitors are rare earth metal cationic (REM) and zinc compounds, which have received much of attention over the past two decades. A comparative study on the corrosion inhibition caused by rare earth metal cations, Ce3+, Pr3+, La3+ and Zn2+ cations on the alloy was done. Cathodic polarization showed that these inhibitor ions suppress the oxygen reduction reaction (ORR) to varying extents with Zn2+ providing the best inhibition. Pr3+ exhibited windows of concentration (100-300 ppm) in which the corrosion rate is minimum; similar to the Ce3+ cation. Scanning Electron Microscopy (SEM) studies showed that the mechanism of inhibition of the Pr3+ ion is also similar to that of the Ce3+ ion. Potentiodynamic polarization experiments after 30 min immersion time showed greatest suppression of oxygen reduction reaction in neutral chloride solutions (pH 7), which reached a maximum at a Zn2+ ion concentration of 5 mM. Anodic polarization experiments after 30 min immersion time, showed no anodic inhibition by the inhibitor in any concentration (0.1 mM - 10 mM) and at any pH. However, anodic polarization of samples immersed after longer immersion times (upto 4 days) in mildly acidic Zn2+ (pH 4) solutions showed significant reduction in anodic kinetics indicating that zinc also acts as a “slow anodic inhibitor”. In contrast to the polarization experiments, coupons exposed to inhibited acidic solutions at pH 4 showed complete suppression of dissolution of Al2CuMg particles compared to zinc-free solutions in the SEM studies. Samples exposed in pH 4 Zn2+-bearing solution exhibited highest polarization resistance which was also observed to increase with time. In deaerated solutions, the inhibition by Zn2+ at pH 4 is not observed as strongly. The ability to make the interfacial electrolyte

  13. Changes of Serum rT3 and Cortisol Levels in Patients with Major Depression Caused by Stress%压力型抑郁症的患者血清rT3和Cortisol的变化研究

    Institute of Scientific and Technical Information of China (English)

    王磊; 立彦

    2016-01-01

    目的 研究探讨反-三碘甲状腺原氨酸(rT3)和皮质醇(Cortisol)在由于压力引发抑郁症患者中的临床应用.方法 本研究纳入39例由于压力引发的抑郁症患者(研究组),以及40例正常体检者(对照组),使用化学发光免疫分析法检测甲状腺功能以及肾上腺功能激素,包括促甲状腺素(TSH)、三碘甲状腺原氨酸(T3)、四碘甲状腺原氨酸(T4)、反-三碘甲状腺原氨酸(rT3)以及皮质醇(Cortisol)含量,并进行比较分析;同时根据病程对研究组进行分组比较.结果 研究组血清rT3以及Cortisol水平显著高于正常对照组(P<0.01),而根据病程的不同,其T3、rT3水平也存在显著差异(P<0.05).结论 压力型抑郁症会导致患者甲状腺以及肾上腺功能异常.

  14. Effects of lipoic acid on lipolysis in 3T3-L1 adipocytes[S

    Science.gov (United States)

    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

  15. Combined effects of genistein, quercetin, and resveratrol in human and 3T3-L1 adipocytes.

    Science.gov (United States)

    Park, Hea Jin; Yang, Jeong-Yeh; Ambati, Suresh; Della-Fera, Mary Anne; Hausman, Dorothy B; Rayalam, Srujana; Baile, Clifton A

    2008-12-01

    The natural compounds genistein (G), quercetin (Q), and resveratrol (R) have been reported to each exhibit anti-adipogenic activities in adipocytes and antiproliferative and pro-apoptotic activities in several cell types. We studied the combined effects of G, Q, and R on adipogenesis and apoptosis in primary human adipocytes (HAs) and 3T3-L1 murine adipocyte (MAs). Combined treatment with 6.25 microM G, 12.5 microM Q, and 12.5 microM R during the 14-day differentiation period caused an enhanced inhibition of lipid accumulation in maturing HAs that was greater than the responses to individual compounds and to the calculated additive response. Glycerol 3-phosphate dehydrogenase activity, a marker of late adipocyte differentiation, was decreased markedly in HAs treated with the combination of G+Q+R. In addition, combined treatment with 50 microM G, 100 microM Q, and 100 microM R for 3 days decreased cell viability and induced apoptosis in early- and mid- phase maturing and lipid-filled mature HAs. In contrast, no compound alone induced apoptosis. Oil Red O stain and Hoechst 33342 stain were performed to confirm the effects on lipid accumulation and apoptosis, respectively. We also determined whether MAs responded to the combination treatment similarly to HAs. As in HAs, G+Q+R treatment decreased lipid accumulation in maturing MAs and increased apoptosis in pre- and lipid-filled mature MAs more than the responses to G, Q, and R when used separately. These results show that lower concentrations of combined treatments with several natural compounds may be useful for treatments for obesity through the suppression of adipogenesis and enhanced adipocyte apoptosis.

  16. Inositol hexakisphosphate inhibits mineralization of MC3T3-E1 osteoblast cultures.

    Science.gov (United States)

    Addison, William N; McKee, Marc D

    2010-04-01

    Inositol hexakisphosphate (IP6, phytic acid) is an endogenous compound present in mammalian cells and tissues. Differentially phosphorylated forms of inositol are well-documented to have important roles in signal transduction, cell proliferation and differentiation, and IP6 in particular has been suggested to inhibit soft tissue calcification (specifically renal and vascular calcification) by binding extracellularly to calcium oxalate and calcium phosphate crystals. However, the effects of IP6 on bone mineralization are largely unknown. In this study, we used MC3T3-E1 osteoblast cultures to examine the effects of exogenous IP6 on osteoblast function and matrix mineralization. IP6 at physiologic concentrations caused a dose-dependent inhibition of mineralization without affecting cell viability, proliferation or collagen deposition. Osteoblast differentiation markers, including tissue-nonspecific alkaline phosphatase activity, bone sialoprotein and osteocalcin mRNA levels, were not adversely affected by IP6 treatment. On the other hand, IP6 markedly increased protein and mRNA levels of osteopontin, a potent inhibitor of crystal growth and matrix mineralization. Inositol alone (without phosphate), as well as inositol hexakis-sulphate, a compound with a high negative charge similar to IP6, had no effect on mineralization or osteopontin induction. Binding of IP6 to mineral crystals from the osteoblast cultures, as well as to synthetic hydroxyapatite crystals, was confirmed by a colorimetric assay for IP6. In summary, IP6 inhibits mineralization of osteoblast cultures by binding to growing crystals through negatively charged phosphate groups and by induction of inhibitory osteopontin expression. These data suggest that IP6 may regulate physiologic bone mineralization by directly acting extracellularly, and by serving as a specific signal at the cellular level for the regulation of osteopontin gene expression.

  17. Suppressive actions of eicosapentaenoic acid on lipid droplet formation in 3T3-L1 adipocytes

    Directory of Open Access Journals (Sweden)

    Sinclair Andrew J

    2010-06-01

    Full Text Available Abstract Background Lipid droplet (LD formation and size regulation reflects both lipid influx and efflux, and is central in the regulation of adipocyte metabolism, including adipokine secretion. The length and degree of dietary fatty acid (FA unsaturation is implicated in LD formation and regulation in adipocytes. The aims of this study were to establish the impact of eicosapentaenoic acid (EPA; C20:5n-3 in comparison to SFA (STA; stearic acid, C18:0 and MUFA (OLA; oleic acid, C18:1n-9 on 3T3-L1 adipocyte LD formation, regulation of genes central to LD function and adipokine responsiveness. Cells were supplemented with 100 μM FA during 7-day differentiation. Results EPA markedly reduced LD size and total lipid accumulation, suppressing PPARγ, Cidea and D9D/SCD1 genes, distinct from other treatments. These changes were independent of alterations of lipolytic genes, as both EPA and STA similarly elevated LPL and HSL gene expressions. In response to acute lipopolysaccharide exposure, EPA-differentiated adipocytes had distinct improvement in inflammatory response shown by reduction in monocyte chemoattractant protein-1 and interleukin-6 and elevation in adiponectin and leptin gene expressions. Conclusions This study demonstrates that EPA differentially modulates adipogenesis and lipid accumulation to suppress LD formation and size. This may be due to suppressed gene expression of key proteins closely associated with LD function. Further analysis is required to determine if EPA exerts a similar influence on LD formation and regulation in-vivo.

  18. The Effects of Topical Application of Thyroid Hormone (Liothyronine, T3 on Skin Wounds in Diabetic Wistar Rats

    Directory of Open Access Journals (Sweden)

    Mahmoud Ali Kaykhaei

    2016-04-01

    Full Text Available Background Efficient treatment of skin ulcers, a leading cause of substantial number of morbidities among diabetic patients, is a subject of matter. Objectives Since current therapies are partially effective and/or expensive and topical liothyronine (T3 was shown to get faster wound healing in mice, the present study was designed to assess its effectiveness in diabetic male rats. Materials and Methods In this experimental study, 30 male wistar rats with mean weight of 242 g were randomly assigned into control (group C (n = 10 and diabetic (n = 20 groups. Diabetes was induced by intraperitoneal (i.p. injection of streptozotocin. Diabetic rats were randomly subdivided into two groups: diabetic T3 group (group A which was treated with 150 ng/day topical T3 and diabetic placebo group (group B received vehicle. Full thickness wound on dorsum was created in each rat (1 cm2. Photographs were taken at baseline, fourth and tenth day to analyze changes in surface areas of wounds. Results Results obtained from the present study showed that baseline surface areas of wounds were similar in all groups. Conversely, wound contraction was significantly better in T3 group in fourth and tenth days compared to placebo group, (P = 0.001, P < 0.00001. Moreover, wound healing was impaired in diabetic placebo group compared to other groups (all P < 0.05. Conclusions This study revealed that topical T3 administration is an effective measure for treatment of ulcers in diabetic male rats.

  19. Troponin T3 regulates nuclear localization of the calcium channel Cavβ1a subunit in skeletal muscle.

    Science.gov (United States)

    Zhang, Tan; Taylor, Jackson; Jiang, Yang; Pereyra, Andrea S; Messi, Maria Laura; Wang, Zhong-Min; Hereñú, Claudia; Delbono, Osvaldo

    2015-08-15

    The voltage-gated calcium channel (Cav) β1a subunit (Cavβ1a) plays an important role in excitation-contraction coupling (ECC), a process in the myoplasm that leads to muscle-force generation. Recently, we discovered that the Cavβ1a subunit travels to the nucleus of skeletal muscle cells where it helps to regulate gene transcription. To determine how it travels to the nucleus, we performed a yeast two-hybrid screening of the mouse fast skeletal muscle cDNA library and identified an interaction with troponin T3 (TnT3), which we subsequently confirmed by co-immunoprecipitation and co-localization assays in mouse skeletal muscle in vivo and in cultured C2C12 muscle cells. Interacting domains were mapped to the leucine zipper domain in TnT3 COOH-terminus (160-244 aa) and Cavβ1a NH2-terminus (1-99 aa), respectively. The double fluorescence assay in C2C12 cells co-expressing TnT3/DsRed and Cavβ1a/YFP shows that TnT3 facilitates Cavβ1a nuclear recruitment, suggesting that the two proteins play a heretofore unknown role during early muscle differentiation in addition to their classical role in ECC regulation.

  20. Autism phenotypes in ZnT3 null mice: Involvement of zinc dyshomeostasis, MMP-9 activation and BDNF upregulation.

    Science.gov (United States)

    Yoo, Min Heui; Kim, Tae-Youn; Yoon, Young Hee; Koh, Jae-Young

    2016-06-29

    To investigate the role of synaptic zinc in the ASD pathogenesis, we examined zinc transporter 3 (ZnT3) null mice. At 4-5 weeks of age, male but not female ZnT3 null mice exhibited autistic-like behaviors. Cortical volume and neurite density were significantly greater in male ZnT3 null mice than in WT mice. In male ZnT3 null mice, consistent with enhanced neurotrophic stimuli, the level of BDNF as well as activity of MMP-9 was increased. Consistent with known roles for MMPs in BDNF upregulation, 2.5-week treatment with minocycline, an MMP inhibitor, significantly attenuated BDNF levels as well as megalencephaly and autistic-like behaviors. Although the ZnT3 null state removed synaptic zinc, it rather increased free zinc in the cytosol of brain cells, which appeared to increase MMP-9 activity and BDNF levels. The present results suggest that zinc dyshomeostasis during the critical period of brain development may be a possible contributing mechanism for ASD.

  1. Effect of T(3) treatment and food ration on hepatic deiodination and conjugation of thyroid hormones in rainbow trout, Oncorhynchus mykiss.

    Science.gov (United States)

    Finnson, K W; Eales, J G

    1999-09-01

    We studied the 7-day effects of 3,5,3'-triiodothyronine (T(3)) hyperthyroidism (induced by 12 ppm T(3) in food) and food ration (0, 0.5, or 2% body weight/day) on in vitro hepatic glucuronidation, sulfation, and deiodination of thyroxine (T(4)), T(3), and 3,3', 5'-triiodothyronine (rT(3)). T(3) treatment doubled plasma T(3) with no change in plasma T(4), depressed hepatic low-K(m) (1 nM) outer-ring deiodination (ORD) of T(4), induced low-K(m) (1 nM) inner-ring deiodination (IRD) of both T(4) and T(3) but did not alter high-K(m) (1 microM) rT(3)ORD, glucuronidation, or sulfation of T(4), T(3), or rT(3). Plasma T(4) levels were greater for 0 and 2% rations than for a 0.5% ration. Fasting decreased low-K(m) T(4)ORD activity and increased high-K(m) rT(3)ORD activity but did not alter T(4)IRD or T(3)IRD activities. T(4), T(3), and rT(3) glucuronidation were greater for 0 and 0.5% rations than for a 2% ration. T(3) glucuronidation was greater for a 0.5% ration than for a 0% ration. T(3) and rT(3) sulfation were greater for a 2% ration than for a 0 or a 0.5% ration; ration did not change T(4) sulfation. We conclude that (i) modest experimental T(3) hyperthyroidism induces T(3) autoregulation by adjusting hepatic low-K(m) ORD and IRD activities but not high-K(m) rT(3)ORD or conjugation activities; (ii) in contrast, ration level changes both deiodination and conjugation pathways, suggesting that the response to ration does not solely reflect altered T(3) production; (iii) deiodination and conjugation appear complementary in regulating thyroidal status in response to ration; and (iv) high-K(m) rT(3)ORD in trout differs from rat type I deiodination in that it does not respond to T(3) hyperthyroidism and it increases, rather than decreases, its activity during fasting. Copyright 1999 Academic Press.

  2. 2024­T3铝合金在模拟海洋大气环境中的腐蚀行为%Corrosion behavior of 2024­T3 aluminum alloy in simulated marine atmospheric environment

    Institute of Scientific and Technical Information of China (English)

    刘艳洁; 王振尧; 柯伟

    2013-01-01

    The corrosion process of 2024­T3 aluminum alloy in marine atmospheric environment was simulated by a cycling salt spraying test. The corrosion evolution of 2024­T3 aluminum alloy as a function of corrosion time (117, 242, 362, 487, 598 h) was investigated by corrosion mass loss test, scanning electron microscopy (SEM), energy­dispersive spectroscopy (EDS) and electrochemical techniques. Consequently, corrosion kinetics, the morphologies of corrosion products and pits, the composition of corrosion products and the electrochemical characters of rust layers were obtained. The influence of the rust layers on the atmospheric corrosion of 2024­T3 aluminum alloy was studied. The results of corrosion kinetics analysis show that the rust layers can well protect 2024­T3 aluminum alloy from corrosion. The results of electrochemical measurement further illustrate that the protective effect of the rust layers is first enhanced, and then weakened, at last enhanced slightly again as the corrosion time increases.%  通过循环盐雾腐蚀实验,模拟2024­T3铝合金在海洋大气环境中的腐蚀过程。采用腐蚀质量损失测试、扫描电镜(SEM)、X射线能谱(EDS)和电化学技术分别对腐蚀117、242、362、487和598 h的2024­T3铝合金试样进行测试分析,得到腐蚀动力学、腐蚀产物和点蚀坑的形貌、腐蚀产物的成分以及表面锈层的电化学特性,研究锈层对2024­T3铝合金大气腐蚀的影响。动力学分析表明,腐蚀过程中2024­T3铝合金的表面形成了具有较好保护性的锈层;电化学测试结果表明,锈层的保护性呈现随腐蚀时间的延长先增强后减弱然后再略增强的变化过程。

  3. 2012 ETA Guidelines: The Use of L-T4 + L-T3 in the Treatment of Hypothyroidism

    DEFF Research Database (Denmark)

    Nygaard, Birte

    2012-01-01

    Background: Data suggest symptoms of hypothyroidism persist in 5–10% of levothyroxine (L-T4)-treated hypothyroid patients with normal serum thyrotrophin (TSH). The use of L-T4 + liothyronine (L-T3) combination therapy in such patients is controversial. The ETA nominated a task force to review....... There is insufficient evidence that L-T4 + L-T3 combination therapy is better than L-T4 monotherapy, and it is recommended that L-T4 monotherapy remains the standard treatment of hypothyroidism. L-T4 + L-T3 combination therapy might be considered as an experimental approach in compliant L-T4-treated hypothyroid...

  4. Cloning and sequencing of murine T3 gamma cDNA from a subtractive cDNA library.

    Science.gov (United States)

    Haser, W G; Saito, H; Koyama, T; Tonegawa, S

    1987-10-01

    The coding sequences of the murine and human T3 gamma chains are of identical length (182 amino acids) and contain a remarkable conservation of residues. The most striking observation is the high degree of homology between the murine and human cytosolic domains (89%), suggesting that the effector function of the T3 complex may be extremely similar or identical within human and murine lymphocytes. Both murine and human T lymphocytes can express two T3 gamma mRNA transcripts, suggesting that a second polyadenylation signal is present downstream. A poly(A) tail is not found in the 3' untranslated region of the murine gamma presented here, indicating that the murine clones analyzed represent mRNA generated by reading through the overlapping poly(A) signals at position 850-860 and possibly terminating at a position that would produce the 1.0 kb transcript.

  5. [The relation between the low T3 syndrome in the clinical course of myocardial infarction and heart failure].

    Science.gov (United States)

    Frączek, Magdalena Maria; Gackowski, Andrzej; Przybylik-Mazurek, Elwira; Nessler, Jadwiga

    2016-06-01

    It has been proven that either excess or deficiency of thyroid hormones has harmful influence on the cardiovascular system function. On the other hand, severe systemic conditions like myocardial infarction or severe heart failure may affect thyroid hormones secretion and their peripheral conversion, leading to low T3 syndrome. Amongst many mechanisms causing T4 to T3 conversion disturbances, important role plays decreased activity of D1 deiodinase and increased activity of D3 deiodinase. The animal research confirmed that thyroid hormones influence cardiomiocytes phenotype and morphology. They inhibit inflammation, apoptosis and cardiac remodelling after myocardial infarction. It was also proven that free triiodothyronine similarly to brain natriuretic peptide predict long-term prognosis in chronic and acute heart failure patients. Potential influence of low T3 syndrome on the course of myocardial infarction and heart failure may have significant impact on the future research on individualization of myocardial infarction and heart failure treatment depending on patient's thyroid status.

  6. Green tea polyphenol (-)-epigallocatechin gallate suppressed the differentiation of murine osteoblastic MC3T3-E1 cells.

    Science.gov (United States)

    Kamon, Masayoshi; Zhao, Ran; Sakamoto, Kazuichi

    2009-12-16

    Recently, various physiological effects of the tea polyphenol catechin for alleviating diseases such as cancer, arteriosclerosis, hyperlipidaemia and osteoporosis have been reported. However, the physiological effect of catechin on bone metabolism remains unclear. We examined the physiological effect of EGCG [(-)-epigallocatechin-3-gallate], which is the main component of green tea catechin, on osteoblast development using the precursor cell line of osteoblasts, MC3T3-E1, and co-culture of the osteoblasts from mouse newborn calvaria and mouse bone marrow cells. Although EGCG did not affect the viability and proliferation of MC3T3-E1 cells, EGCG inhibited the osteoblast differentiation. Furthermore, EGCG did not affect the mineralization of differentiated MC3T3-E1 cells, and reduced osteoclast formation in co-culture. These results suggest that EGCG can effectively suppress bone resorption, and can be used as an effective medicine in the treatment of the symptoms of osteoporosis.

  7. Changes of Serum Trace Elements, AFP, CEA, SF, T3, T4 and IGF- Ⅱ in Different Periods of Rat Liver Cancer

    Institute of Scientific and Technical Information of China (English)

    Hong-xu Zhang; Dan-dan Liu; Bai-jie Jin; Ya-wei Wang; Qj Liu; Ru-bing Duan; Peng Zhao; Ming-xia Ma

    2011-01-01

    Objective:Based on liver cancer model built in SD rats,the contents of trace elements (Cu,Fe,Zn,Ca and Mg),AFP,CEA,SF,TH and IGF-Ⅱ in serum were measured at different stages to explore the molecular changes during the rat liver cancer development.Methods:The SD rat liver cancer model was built by using diethylnitrosamine (DENA) as the mutagen.During 16 weeks of DENA gavage,blood samples were taken in the 14th,28th,56th,77th,105th and 112th days respectively after the first day of gavage with DENA,then the contents of five trace elements (Cu,Fe,Zn,Ca and Mg),T3,T4,IGF-Ⅱ,AFP,CEA and SF in serum were determined.Results:During the development of the rat liver cancer,in the test group,the Cu content significantly increased in serum,while the contents of Fe,Zn and Ca significantly decreased.The content of Mg showed no significant change.AFP and CEA of the test group showed same expression level with the control group; while the content of SF was lower than that of the control group when cancerization appeared.T3 and T4 increased at the first stage and then went down,and the content of IGF-Ⅱ was always high.Conclusion:Cu,Fe,Zn,Ca,T3,T4,SF and IGF-Ⅱ are closely related to the development of liver cancer.The changes of their contents in the development of cancer could enlighten the researches on cancer pathogenesis and prevention.

  8. Effect of Tumor Necrosis Factor-α on Resistin Expression in 3T3-L1 Adipocytes and Its Mechanism

    Institute of Scientific and Technical Information of China (English)

    2005-01-01

    In order to investigate the effect of tumor necrosis factor-α (TNFα) on resistin expression in 3T3-L1 adipocytes, and further explore its mechanisms, the differentiated 3T3-L1 adipocytes were incubated with 0, 1, 10, 100 ng/mL TNFα respectively for 24 h, and then the expression of resistin was determined. The differentiated 3T3-L1 adipocytes were incubated with 100 ng/mL TNFα for 3, 6, 24 h respectively, and then the expression of resistin mRNA was analyzed.3T3-L1 adipocytes were induced to differentiate into mature adipocytes. The cells were randomly divided into 4 groups for culture. In the control group, no drugs were added. Cells of TNFα group were treated with 100 ng/mL TNFα. In Ro-31-8220 group, 5μmol/L protein kinase C inhibitor Ro-31-8220 was added. With TNFα+Ro-31-8220 group, 100 ng/mL TNFα were added 1 h after the addition of 5 μmol/L Ro-31-8220. All adipocytes were cultured for 24 h. Reverse transcriptionpolymerase chain reaction (RT-PCR) and Western blotting were employed to detect the expression of resistin gene. Our results showed that resistin protein and mRNA in 3T3-L1 adipocytes were inhibited by TNFα at different concentrations (P<0.01), and the inhibitory effect increased with the concentration (P<0.01). At the same concentrations, the inhibitory effect increased with time (P <0.01). Ro-31-8220 could inhibit its expression and the inhibitive effect remained unchanged with addition of TNFα(P>0.05). It was concluded that TNFα could inhibit the expression of resistin in 3T3-L1 adipocytes. The mechanism may be that the expression of resistin is partly controlled by protein kinase C signal conduction pathway.

  9. Intervalos de referencia para concentraciones séricas de T3 y T4. Estudio preliminar

    Directory of Open Access Journals (Sweden)

    Cecilia Miranda Pantoja

    2013-12-01

    Full Text Available Fundamento: la determinación cuantitativa de las hormonas tiroideas T3 y T4 reviste gran importancia en el diagnóstico y la evaluación del hipertiroidismo, en especial del hipertiroidismo aislado causado por T3.Objetivo: establecer los intervalos de referencia de T3 y T4 en el laboratorio de medicina nuclear del Hospital General Universitario Dr. Gustavo Aldereguía Lima de Cienfuegos. Métodos: estudio descriptivo y prospectivo realizado mediante el método de radioinmunoanálisis, competencia que se establece entre la T3 y T4 sin marcar, y la marcada por un número limitado de los sitios de unión del anticuerpo específico. Al hacer reaccionar una cantidad fija de trazador y anticuerpo con diferentes cantidades del ligando sin marcar, la cantidad de trazador unido por el anticuerpo será inversamente proporcional a la concentración del ligando sin marcar. Resultados: los valores obtenidos se describen según una distribución gaussiana (media aritmética = 117, desviación estándar =31 para T4; media aritmética = 2,64, desviación estándar = 0,64 para T3, comprobado mediante un test de Chi cuadrado. Los rangos de valores normales obtenidos fueron de de 55 – 178 nmol/L y 1,4 – 3,9 nmol/L para T4 y T3 respectivamente. Conclusiones: los intervalos de referencia obtenidos resultaron más amplios que los propuestos por el productor, sobre todo en el caso de T4.

  10. Regulation of glucose transport in the NIH 3T3 L1 preadipocyte cell line by TCDD.

    OpenAIRE

    1994-01-01

    This study examined the changes in cellular glucose uptake induced by 2,3,7,8 tetrachlorodibenzo-p-dioxin (TCDD) as measured by quantification of intracellular radioactivity in the NIH 3T3 L1 preadipocyte cell line after a 30-minute incubation with the non-metabolizable radioactive analogue of glucose, 3-O-methyl-D-[1-3H] glucose. Treatment of differentiated NIH 3T3 L1 cells with TCDD produced a time- and dose-dependent decrease in the cellular uptake of glucose. Treatment of cells for 3 hr w...

  11. Inhibition of 3T3-L1 adipocyte differentiation by expression of acyl-CoA-binding protein antisense RNA

    DEFF Research Database (Denmark)

    Mandrup, S; Sorensen, R V; Helledie, T

    1998-01-01

    adipocyte differentiation. In this report we describe the effects of expression of high levels of ACBP antisense RNA on the differentiation of 3T3-L1 cells. Pools of 3T3-L1 cells transfected with vectors expressing ACBP antisense RNA showed significantly less lipid accumulation as compared with cells...... of the adipogenic transcription factors peroxisome proliferator-activated receptor gamma and CCAAT/enhancer-binding protein alpha as well as several adipocyte-specific genes was significantly delayed and reduced. The adipogenic potential of antisense-expressing cells was partially restored by transfection...

  12. Soy pinitol acts partly as an insulin sensitizer or insulin mediator in 3T3-L1 preadipocytes

    OpenAIRE

    Do, Gyeong-Min; Choi, Myung-Sook; Kim, Hye-Jin; Woo, Myung-Nam; Lee, Mi-Kyung; Jeon, Seon-Min

    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...

  13. Polyphosphates inhibit extracellular matrix mineralization in MC3T3-E1 osteoblast cultures.

    Science.gov (United States)

    Hoac, Betty; Kiffer-Moreira, Tina; Millán, José Luis; McKee, Marc D

    2013-04-01

    Studies on various compounds of inorganic phosphate, as well as on organic phosphate added by post-translational phosphorylation of proteins, all demonstrate a central role for phosphate in biomineralization processes. Inorganic polyphosphates are chains of orthophosphates linked by phosphoanhydride bonds that can be up to hundreds of orthophosphates in length. The role of polyphosphates in mammalian systems, where they are ubiquitous in cells, tissues and bodily fluids, and are at particularly high levels in osteoblasts, is not well understood. In cell-free systems, polyphosphates inhibit hydroxyapatite nucleation, crystal formation and growth, and solubility. In animal studies, polyphosphate injections inhibit induced ectopic calcification. While recent work has proposed an integrated view of polyphosphate function in bone, little experimental data for bone are available. Here we demonstrate in osteoblast cultures producing an abundant collagenous matrix that normally show robust mineralization, that two polyphosphates (PolyP5 and PolyP65, polyphosphates of 5 and 65 phosphate residues in length) are potent mineralization inhibitors. Twelve-day MC3T3-E1 osteoblast cultures with added ascorbic acid (for collagen matrix assembly) and β-glycerophosphate (a source of phosphate for mineralization) were treated with either PolyP5 or PolyP65. Von Kossa staining and calcium quantification revealed that mineralization was inhibited in a dose-dependent manner by both polyphosphates, with complete mineralization inhibition at 10μM. Cell proliferation and collagen assembly were unaffected by polyphosphate treatment, indicating that polyphosphate inhibition of mineralization results not from cell and matrix effects but from direct inhibition of mineralization. This was confirmed by showing that PolyP5 and PolyP65 bound to synthetic hydroxyapatite in a concentration-dependent manner. Tissue-nonspecific alkaline phosphatase (TNAP, ALPL) efficiently hydrolyzed the two PolyPs as

  14. Functional cross-talk between angiotensin II and epidermal growth factor receptors in NIH3T3 fibroblasts.

    Science.gov (United States)

    De Paolis, Paola; Porcellini, Antonio; Savoia, Carmine; Lombardi, Alessia; Gigante, Bruna; Frati, Giacomo; Rubattu, Speranza; Musumeci, Beatrice; Volpe, Massimo

    2002-04-01

    The main angiotensin (Ang) II subtype receptors (AT1R and AT2R) are involved in cellular growth processes and exert functionally antagonistic effects. To characterize the mechanisms by which Ang II receptors influence growth, by investigating the interactions between Ang II subtype receptors and epidermal growth factor (EGF) receptors on mitogen-activated protein kinase (MAPK) pathway activation. The experiments were performed using a mouse fibroblast cell line, NIH3T3, by transient co-transfection with rat AT1R or AT2R expression vectors, or both. Extracellular-signal-regulated kinase (ERK)1/2 phosphorylation was analysed by western blot and the ERK activity was evaluated using PathDetect, an in-vivo signal transduction pathway trans-reporting system. Selective Ang II receptor antagonists (losartan for AT1R and PD123319 for AT2R) were used to investigate the contributions of each receptor to the response observed. Our data show that, in this cellular model, both Ang II receptors phosphorylate ERK1/2. However, in the cells expressing AT1R, the EGF-induced MAPK pathway was enhanced in the presence of Ang II in a synergistic fashion. In contrast, a reduction of EGF-induced MAPK activation was observed in the cells expressing AT2R. In cells expressing both Ang II subtype receptors, Ang II promoted an enhancement of EGF-induced MAPK activation. However, in the presence of the AT1R antagonist, losartan, the effect of EGF was reduced. These data indicate the existence of an opposite cross-talk of AT1R and AT2R with EGF receptors, and suggest a complex functional interaction between these pathways in the regulation of cellular growth processes.

  15. 小檗碱对3T3-L1脂肪细胞脂联素表达的影响%Effects of berberine on adiponectin mRNA expression in 3T3-L1 adipocyte

    Institute of Scientific and Technical Information of China (English)

    谷卫; 曾文衡; 胡海英

    2005-01-01

    目的:探讨小檗碱和胰岛素对3T3-L1脂肪细胞脂联素表达的影响.方法:检测分别经小檗碱及胰岛素处理后3T3-L1脂肪细胞脂联素表达水平改变,以β-actin为内对照,半定量法RT-PCR测定脂联素mRNA表达.结果:经小檗碱(10 μmol·L-1)干预后3T3-L1脂肪细胞脂联素表达显著增加(P<0.05),加入胰岛素后脂联素的表达受到抑制,高浓度胰岛素有显著抑制作用(P<0.05). 结论:体外实验中,小檗碱使3T3-L1脂肪细胞脂联素的表达增加,而胰岛素可抑制小檗碱增加脂联素表达的作用.

  16. Internal thermal performance measurement of electronic elements in space by T3Ster%使用T3Ster对宇航电子元器件内部热特性的测量

    Institute of Scientific and Technical Information of China (English)

    张立; 汪新刚; 崔福利

    2011-01-01

    文章介绍了使用MicReD公司的热测试仪T3Ster测量元器件内部热特性的方法.T3Ster测试仪可以测试各类IC、LED、散热器、热管等电子器件的热特性以及PCB、导热材料等的热阻、热容及导热系数、接触热阻等热特性.使用T3Ster测试仪对某航天器用电子元器件内部热特性进行了测量,并与器件资料中的热特性数据进行比对,二者相对误差为0.07%,验证了T3Ster测试仪具有测试高可靠度要求的宇航级电子元器件热特性的能力.为宇航电子元器件的热设计与热分析提供重要的试验依据.

  17. Thyroxine (T 4-RIA) and triiodothyronine (T 3-RIA) serum levels in sheep fed on Leucaena Leucocephala; Niveis sericos de tiroxina (T4-RIA) e triiodotinonina (T3-RIA) em ovinos alimentados cm Leucaena Leucocephala LAM

    Energy Technology Data Exchange (ETDEWEB)

    Pessoa, J.M.; Rodriguez, N.M. [Minas Gerais Univ., Belo Horizonte, MG (Brazil). Escola de Veterinaria; Cardoso, W.M. [Maranhao Univ., Sao Luis, MA (Brazil). Escola de Medicina Veterinaria; Velez, C.E.S.

    1988-12-31

    The aim of this work is to study the toxicity of Leucaena leucocephala on thyroxine and triiodothyronine serum concentration. Results indicate that ovine T 4 and T 3 serum levels do not decrease with Leucaena leucocephala feeding, sohen proreided for 41 days. (author). 10 refs, 1 tab.

  18. Possibilities and limitations of combined radio-surgical treatment of large orofacial T3/T4-tumors. Moeglichkeiten und Grenzen der kombiniert radiotherapeutisch-chirurgischen Behandlung grosser orofazialer T3-/T4-Tumoren

    Energy Technology Data Exchange (ETDEWEB)

    Kaercher, K.H. (Universitaetsklinik fuer Strahlentherapie und Strahlenbiologie, Vienna (Austria)); Kaercher, H. (Abteilung fuer Mund-, Kiefer- und Gesichtschirurgie, Univ. Graz (Austria))

    1993-05-01

    The possibilities, limits and good cosmetic and functional results of combined radiotherapy-surgical treatment of large orofacial tumors (T3) are prescribed. The pre- and postoperative splitted radiotherapy with operation in between is favoured. The radical en-bloc-resection with radical dissection is the surgical therapy of choice. The defects have been reconstructed with osteomusculocutaneous flaps. (orig.)

  19. Specific Labeling of Mouse 3T3-L1 Preadipocyte Cell Line with Green Fluorescent Protein%小鼠3T3-L1前脂肪细胞系的特异性标记

    Institute of Scientific and Technical Information of China (English)

    张崇本; 张晓兰; 李成健; 成俊英; 吴显荣

    2004-01-01

    A vector of paP2-promoter-EGFP was constructed and introduced into mouse 3T3-L1 preadipocyte cells, a cell line derived from mouse Swiss3T3 cells that were isolated from mouse embryo, to make the cells labelled with enhanced green fluorescent protein (EGFP) whose expression was controlled by the promoter of adipose-specific gene aP2. The cells were then induced to differentiate and the expression of aP2 was detected by EGFP-microscopy and RT-PCR assays. The EGFP gene was transferred into the mouse 3T3-L1 preadipocyte cells, and EGFP expression and lipid accumulation were observed during differentiation. The expression of aP2 was stable and similar to the expression of EGFP. A preadipocyte cell line expressing EGFP was obtained under the control of the promoter of adipocyte-specific expression gene aP2, and the preadipocyte cell line was specifically labelled. The cell line provides a powerful approach for the research of adipocyte differentiation and for the screening of anti-obesity and anti-diabetes drugs.%用增强绿色荧光蛋白特异性标记小鼠3T3-L1前脂肪细胞系.构建pap2-promoter-EGFP载体,电穿孔转染小鼠3T3-L1前脂肪细胞,显微荧光观察和RT-PCR确认aP2基因的内源表达.EGFP基因转入3T3-L1前脂肪细胞,观察到细胞分化过程中EGFP表达和脂肪积累.RT-PCR分析表明,EGFP代表了稳定而真实的aP2基因的内源性表达.建立了由脂肪组织特异表达基因aP2的表达控制的EGFP标记的小鼠3T3-L1前脂肪细胞系,目前尚未见用同样方法对前脂肪细胞进行特异性标记.该细胞系将为脂肪细胞分化机理研究以及为抗肥胖症和抗糖尿病药物筛选提供有力工具.

  20. Berberine Activates AMPK and Inhibits 3T3-L1 Adipocyte Differentiation%小檗碱激活AMPK抑制3T3-L1脂肪细胞分化

    Institute of Scientific and Technical Information of China (English)

    王宁; 张娟; 建方方; 邓儒元; 唐红菊; 刘赟; 李凤英; 王晓; 周丽斌

    2012-01-01

    目的 探讨小檗碱对3T3-L1脂肪分化的作用是否与激活腺苷酸活化蛋白激酶(AMPK)有关.方法 在3T3-L脂肪细胞分化全程加入小檗碱,以油红O染色检测3T3-L1脂肪细胞胞浆中脂肪的堆积,实时定量PCR检测过氧化物酶体增殖物激活受体γ2(PPARγ2)、CCAAT增强子结合蛋白α(CEBPα)和AMPK的mRNA表达,以Western印迹法检测AMPK和乙酰辅酶A羧化酶(ACC)的磷酸化水平.结果 小檗碱剂量依赖性地抑制3T3-L1脂肪细胞分化,10 μmol/L小檗碱几乎完全抑制胞浆中脂肪的堆积.5 μmol/L小檗碱在脂肪细胞诱导分化1、3、5、7d后均显著降低CEBPα mRNA表达(P<0.05或P<0.01),诱导分化3、5、7d时显著降低PPARγ2的mRNA表达(P<0.05或P<0.01).AMPK的mRNA水平在分化过程中未受小檗碱的明显影响,而小檗碱明显增加其蛋白磷酸化水平,其下游靶基因ACC磷酸化水平也明显增加.结论 小檗碱抑制3T3-L1脂肪细胞的分化可能与其激活AMPK有关.%Objective To investigate whether the effect of berberine ( BBR) on 3T3-L1 adipocyte differentiation is related to AMP activated protein (AMPK) activation. Methods The accumulation of lipid in the cytoplasm of differentiated 3T3-L1 adipocytes was observed by oil red 0 staining. Realtime-PCR was used to detect the mRNA ezpiesBions of PPARγ2, CEBPα, and AMPK. The phosphorylation levels of AMPK and acetyl CoA carboxylase (ACC) were detected by Western blot. Result Berberine inhibited 3T3-L1 adipocyte differentiation in a dose-dependent manner. At the concentration of 10μmol/L berberine, the accumulation of lipid in the cytoplasm of adipocytes was almost inhibited. CEBPa mRNA expression was inhibited by 5μmol/L berberine after 1,3,5, and 7day induction differentiation (P<0.05 or P<0.01) and PPARry2 mRNA expression was decreased by berberine after induction differentiation of 3,5, and 7 day (P<0.05 or P< 0.01). There were no changes of AMPK mRNA level after 3T3-IA cells were incubated with

  1. Effect of Liupao Tea on Glucose and Lipid Metabolism in Palmitate-induced Insulin Resistance 3T3-L1 Adipocytes%六堡茶对胰岛素抵抗3T3-L1脂肪细胞糖脂代谢的影响

    Institute of Scientific and Technical Information of China (English)

    滕翠琴; 刘仲华; 龚受基; 彭雨轩; 马蕊

    2014-01-01

    To investigate the effect of Liupao tea on glucose and lipid metabolism in palmitate-induced insulin resistance 3T3-L1 adipocytes. Glucose intake decreased and insulin resistance was induced in 3T3-L1 adipocytes after incubation in 1mmol/L palmitate for 24 hours. Compared with the model group, the concentrations of glucose and NEFA decreased along with the increase of Liupao tea concentrations in medium. The mRNA expression of key enzyme for glycolysis of glucose (Hexokinase, 6-phosphofructokinase-1, and pyruvatekinase) and the Lipid-Metabolism-Related Enzymes (Sn-glycerol-3-phosphate acyltransferase) significantly increased in the Liupao tea groups comparing with model sample. Meanwhile the glucose transporter 4 mRNA expression was also increased and the acetyl-CoA carboxylase, protein tyrosine phosphatase 1B mRNA expression was decreased. Low doses of Liupao tea reduced carnitine palmitoyl transferase I mRNA expression while high doses significantly raised it. Research proved that Liupao tea were able to increase glucose uptake and activate the glucolipid metabolic pathways.Liupao tea showed the effect of improving insulin resistance induced by palmitate in 3T3 -L1 adipocytes.%采用软脂酸诱导3T3-L1脂肪细胞胰岛素抵抗后,给予不同剂量的六堡茶水提物治疗,探讨六堡茶对胰岛素抵抗3T3-L1脂肪细胞的糖脂代谢的影响。结果表明,1 mmol·L-1的软脂酸对3T3-L1脂肪细胞作用24 h后,葡萄糖摄取量减少,建立胰岛素抵抗模型。六堡茶对模型细胞干预24 h 后,与模型组比较,细胞培养基的残留液中葡萄糖和游离脂肪酸的含量降低;对糖脂代谢相关关键酶(己糖激酶、6-磷酸果糖激酶-1、丙酮酸激酶、甘油-3-磷酸酰基转移酶)和葡萄糖转运子基因表达都有上调作用,并下调乙酰辅酶 A 羧化酶、蛋白酪氨酸磷酸酯酶1B基因表达。低剂量的六堡茶下调肉碱脂酰转移酶-I基因的表达,但是高剂量能

  2. The pathogenicity factor HrpF interacts with HrpA and HrpG to modulate type III secretion system (T3SS) function and t3ss expression in Pseudomonas syringae pv. averrhoi.

    Science.gov (United States)

    Huang, Yi-Chiao; Lin, Yuan-Chuen; Wei, Chia-Fong; Deng, Wen-Ling; Huang, Hsiou-Chen

    2016-09-01

    To ensure the optimal infectivity on contact with host cells, pathogenic Pseudomonas syringae has evolved a complex mechanism to control the expression and construction of the functional type III secretion system (T3SS) that serves as a dominant pathogenicity factor. In this study, we showed that the hrpF gene of P. syringae pv. averrhoi, which is located upstream of hrpG, encodes a T3SS-dependent secreted/translocated protein. Mutation of hrpF leads to the loss of bacterial ability on elicitation of disease symptoms in the host and a hypersensitive response in non-host plants, and the secretion or translocation of the tested T3SS substrates into the bacterial milieu or plant cells. Moreover, overexpression of hrpF in the wild-type results in delayed HR and reduced t3ss expression. The results of protein-protein interactions demonstrate that HrpF interacts directly with HrpG and HrpA in vitro and in vivo, and protein stability assays reveal that HrpF assists HrpA stability in the bacterial cytoplasm, which is reduced by a single amino acid substitution at the 67th lysine residue of HrpF with alanine. Taken together, the data presented here suggest that HrpF has two roles in the assembly of a functional T3SS: one by acting as a negative regulator, possibly involved in the HrpSVG regulation circuit via binding to HrpG, and the other by stabilizing HrpA in the bacterial cytoplasm via HrpF-HrpA interaction prior to the secretion and formation of Hrp pilus on the bacterial surface.

  3. Cell competition in mouse NIH3T3 embryonic fibroblasts is controlled by the activity of Tead family proteins and Myc.

    Science.gov (United States)

    Mamada, Hiroshi; Sato, Takashi; Ota, Mitsunori; Sasaki, Hiroshi

    2015-02-15

    Cell competition is a short-range communication originally observed in Drosophila. Relatively little is known about cell competition in mammals or in non-epithelial cells. Hippo signaling and its downstream transcription factors of the Tead family, control cell proliferation and apoptosis. Here, we established an in vitro model system that shows cell competition in mouse NIH3T3 embryo fibroblast cells. Co-culture of Tead-activity-manipulated cells with normal (wild-type) cells caused cell competition. Cells with reduced Tead activity became losers, whereas cells with increased Tead activity became super-competitors. Tead directly regulated Myc RNA expression, and cells with increased Myc expression also became super-competitors. At low cell density, cell proliferation required both Tead activity and Myc. At high cell density, however, reduction of either Tead activity or Myc was compensated for by an increase in the other, and this increase was sufficient to confer 'winner' activity. Collectively, NIH3T3 cells have cell competition mechanisms similar to those regulated by Yki and Myc in Drosophila. Establishment of this in vitro model system should be useful for analyses of the mechanisms of cell competition in mammals and in fibroblasts. © 2015. Published by The Company of Biologists Ltd.

  4. Soshiho-Tang Aqueous Extract Exerts Antiobesity Effects in High Fat Diet-Fed Mice and Inhibits Adipogenesis in 3T3-L1 Adipocytes

    Directory of Open Access Journals (Sweden)

    Sae-Rom Yoo

    2016-01-01

    Full Text Available Soshiho-tang (SST; sho-saiko-to in Japanese; xiaochaihu-tang in Chinese has generally been used to improve liver fibrosis- and cirrhosis-related symptoms in traditional Korean medicine. Although many studies have investigated the pharmacological properties of SST, its antiobesity effect has not been elucidated. Thus, our present study was carried out to evaluate the antiobesity effect of SST using a high fat diet- (HFD induced obese mouse model and 3T3-L1 adipose cells. C57BL/6J mice were randomly divided into four groups (n=6/group, normal diet (ND, HFD-fed group, and HFD- and SST-fed groups (S200: 200 mg/kg of SST; S600: 600 mg/kg of SST and given HFD with or without SST extract for 8 weeks. 3T3-L1 preadipocytes were differentiated into adipocytes for 8 days with or without SST. In the HFD-fed obese mice, body weight and fat accumulation in adipose tissue were significantly reduced by SST administration. Compared with control-differentiated adipocytes, SST significantly inhibited lipid accumulation by decreasing the triglyceride (TG content and leptin concentration in 3T3-L1 adipocytes. SST also decreased the expression of adipogenesis-related genes including lipoprotein lipase (LPL, fatty acid binding protein 4 (FABP4, CCAAT/enhancer-binding protein-alpha (C/EBP-α, and peroxisome proliferator-activated receptor-gamma (PPAR-γ. Our findings suggest that SST has potential as a nontoxic antiobesity medication.

  5. Case Report of S1Q3T3 Electrocardiographic Abnormality in a Pregnant Asthmatic Patient During Acute Bronchospasm

    Science.gov (United States)

    Arshad, Hafiza; Khan, Rana Rahel; Khaja, Misbahuddin

    2017-01-01

    Patient: Female, 33 Final Diagnosis: S1Q3T3 electrocardiographic abnormality in a pregnant asthmatic during acute bronchospasm Symptoms: Cough • shortness of breath Medication: — Clinical Procedure: EKG Specialty: Pulmonology Objective: Rare co-existance of disease or pathology Background: Asthma is the most common chronic pulmonary disease during pregnancy. Several previous reports have documented reversible electrocardiographic changes during severe acute asthma attacks, including tachycardia, P pulmonale, right bundle branch block, right axis deviation, and ST segment and T wave abnormalities. Case Report: We present the case of a pregnant patient with asthma exacerbation in which acute bronchospasm caused S1Q3T3 abnormality on an electrocardiogram (ECG). The complete workup of ECG findings of S1Q3T3 was negative and correlated with bronchospasm. The S1Q3T3 electrocardiographic abnormality can be seen in acute bronchospasm in pregnant women. The other causes like pulmonary embolism, pneumothorax, acute lung disease, cor pulmonale, and left posterior fascicular block were excluded. Conclusions: Asthma exacerbations are of considerable concern during pregnancy due to their adverse effect on the fetus, and optimization of asthma treatment during pregnancy is vital for achieving good outcomes. Prompt recognition of electrocardiographic abnormality and early treatment can prevent adverse perinatal outcomes. PMID:28144025

  6. Rho family GTP binding proteins are involved in the regulatory volume decrease process in NIH3T3 mouse fibroblasts

    DEFF Research Database (Denmark)

    Pedersen, Stine F; Beisner, Kristine H; Willumsen, Berthe M

    2002-01-01

    The role of Rho GTPases in the regulatory volume decrease (RVD) process following osmotic cell swelling is controversial and has so far only been investigated for the swelling-activated Cl- efflux. We investigated the involvement of RhoA in the RVD process in NIH3T3 mouse fibroblasts, using wild-...

  7. Thyroid-stimulating hormone inhibits adipose triglyceride lipase in 3T3-L1 adipocytes through the PKA pathway.

    Directory of Open Access Journals (Sweden)

    Dongqing Jiang

    Full Text Available Thyroid-stimulating hormone (TSH has been shown to play an important role in the regulation of triglyceride (TG metabolism in adipose tissue. Adipose triglyceride lipase (ATGL is a rate-limiting enzyme controlling the hydrolysis of TG. Thus far, it is unclear whether TSH has a direct effect on the expression of ATGL. Because TSH function is mediated through the TSH receptor (TSHR, TSHR knockout mice (Tshr-/- mice (supplemented with thyroxine were used in this study to determine the effects of TSHR deletion on ATGL expression. These effects were verified in 3T3-L1 adipocytes and potential underlying mechanisms were explored. In the Tshr-/- mice, ATGL expression in epididymal adipose tissue was significantly increased compared with that in Tshr+/+ mice. ATGL expression was observed to increase with the differentiation process of 3T3-L1 preadipocytes. In mature 3T3-L1 adipocytes, TSH significantly suppressed ATGL expression at both the protein and mRNA levels in a dose-dependent manner. Forskolin, which is an activator of adenylate cyclase, suppressed the expression of ATGL in 3T3-L1 adipocytes. The inhibitory effects of TSH on ATGL expression were abolished by H89, which is a protein kinase A (PKA inhibitor. These results indicate that TSH has an inhibitory effect on ATGL expression in mature adipocytes. The associated mechanism is related to PKA activation.

  8. Esophageal motion characteristics in thoracic esophageal cancer: Impact of clinical stage T4 versus stages T1-T3

    Directory of Open Access Journals (Sweden)

    Yuta Kobayashi, MS

    2016-10-01

    Conclusions: The EM and the ITV margins in cT4 were significantly smaller than those in cT1-T3. The NM and the ITV margins of abdominal LNs were much larger than those of cervicothoracic LNs and the esophagus. In clinical radiation therapy planning for esophageal cancer, we should take cT stage into consideration.

  9. Knockdown of LYRM1 rescues insulin resistance and mitochondrial dysfunction induced by FCCP in 3T3-L1 adipocytes.

    Science.gov (United States)

    Zhang, Min; Qin, Zhen-Ying; Dai, Yong-mei; Wang, Yu-Mei; Zhu, Guan-zhong; Zhao, Ya-Ping; Ji, Chen-Bo; Zhu, Jin-Gai; Shi, Chun-Mei; Qiu, Jie; Cao, Xin-Guo; Guo, Xi-Rong

    2014-09-01

    LYR motif-containing 1 (LYRM1) was recently discovered to be involved in adipose tissue homeostasis and obesity-associated insulin resistance. We previously demonstrated that LYRM1 overexpression might contribute to insulin resistance and mitochondrial dysfunction. Additionally, knockdown of LYRM1 enhanced insulin sensitivity and mitochondrial function in 3T3-L1 adipocytes. We investigated whether knockdown of LYRM1 in 3T3-L1 adipocytes could rescue insulin resistance and mitochondrial dysfunction induced by the cyanide p-trifluoromethoxyphenyl-hydrazone (FCCP), a mitochondrion uncoupler, to further ascertain the mechanism by which LYRM1 is involved in obesity-associated insulin resistance. Incubation of 3T3-L1 adipocytes with 1 µM FCCP for 12 h decreased insulin-stimulated glucose uptake, reduced intracellular ATP synthesis, increased intracellular reactive oxygen species (ROS) production, impaired insulin-stimulated Glucose transporter type 4 (GLUT4) translocation, and diminished insulin-stimulated tyrosine phosphorylation of Insulin receptor substrate-1 (IRS-1) and serine phosphorylation of Protein Kinase B (Akt). Knockdown of LYRM1 restored insulin-stimulated glucose uptake, rescued intracellular ATP synthesis, reduced intracellular ROS production, restored insulin-stimulated GLUT4 translocation, and rescued insulin-stimulated tyrosine phosphorylation of IRS-1 and serine phosphorylation of Akt in FCCP-treated 3T3-L1 adipocytes. This study indicates that FCCP-induced mitochondrial dysfunction and insulin resistance are ameliorated by knockdown of LYRM1.

  10. 目前肾细胞癌TNM分期的评价:需要修订T3a期

    Institute of Scientific and Technical Information of China (English)

    叶雄俊; 张小东

    2005-01-01

    近期研究表明,直接侵犯肾上腺的pT3。期肾细胞癌患者生存期比其他pT3。期患者更短,因此建议此类患者应该归入pT4期。作者根据肾周脂肪是否浸润评价pT3a诊断的预后价值。回顾性研究从1975—2000年德国Saarland大学泌尿外科手术的1794例肾细胞癌患者资料,尤其是pT3a期肿瘤(浸润肾周脂肪),以肿瘤特异性生存为主要观察终点,通过单因素和多因素统计学方法分析患者的临床和组织病理学资料。

  11. Metabolic effects of chronic T3 administration in the hypothalamic paraventricular and ventromedial nucleus in male rats

    NARCIS (Netherlands)

    Zhang, Z; Foppen, E; Su, Y; Bisschop, P H; Kalsbeek, A; Fliers, E; Boelen, A

    2016-01-01

    Thyroid hormone is a key regulator of energy metabolism. Apart from its direct effects on peripheral metabolism, thyroid hormone exerts acute metabolic effects via distinct nuclei within the hypothalamus. Recently, we developed a method for chronic and local intra-hypothalamic triiodothyronine (T3)

  12. Effect of cortisol on calpains in the C2C12 and 3T3-L1 cells.

    Science.gov (United States)

    Muthuraman, Pandurangan; Ravikumar, Sambandam; Muthuviveganandavel, Veerappan; Kim, Jongpil

    2014-03-01

    The present study was carried out to understand the effect of cortisol on calpain system in the C2C12 and 3T3-L1 adipocyte cells under co-culture system. Cells were co-cultured by using transwell inserts with a 0.4 μm porous membrane to separate C2C12 and 3T3-L1 preadipocyte cells. Each cell type was grown independently on the transwell plates. Following cell differentiation, inserts containing 3T3-L1 cells were transferred to C2C12 plates. Ten microgram per milliliter of cortisol was added to the medium. Following treatment for 3 days, the cells in the lower well were harvested for analysis. Calpains such as μ-calpain, m-calpain, and calpastatin were selected for the analysis. RT-PCR results indicated the significant increase in the mRNA expression of μ-calpain, m-calpain, and calpastatin. In addition, the confocal microscopical investigation indicated the cortisol treatment increases calpain expression in the C2C12 and 3T3-L1 cells. Taking all these together, cortisol treatment with co-culture system shows most reliable status of calpains expression in the cells, which is quite distinct from one-dimensional monocultured cells.

  13. Piromelatine decreases triglyceride accumulation in insulin resistant 3T3-L1 adipocytes: role of ATGL and HSL.

    Science.gov (United States)

    Wang, Ping-Ping; She, Mei-Hua; He, Ping-Ping; Chen, Wu-Jun; Laudon, Moshe; Xu, Xuan-Xuan; Yin, Wei-Dong

    2013-08-01

    Piromelatine, a novel investigational multimodal sleep medicine, is developed for the treatment of patients with primary and co-morbid insomnia. Piromelatine has been shown to inhibit weight gain and improve insulin sensitivity in high-fat/high-sucrose-fed (HFHS) rats. Considering that piromelatine has also been implicated in lowering of triglyceride levels in HFHS rats, this work elucidated whether this effect involves in the regulation of adipose triglyceride lipase (ATGL) and hormone-sensitive lipase (HSL) in triglyceride (TG) metabolism. In this study, we investigated the effects of piromelatine and MT2 receptors inhibition on TG content, insulin-stimulated glucose uptake, and the expressions of ATGL and HSL in 3T3-L1 adipocytes preincubated in high glucose and high insulin (HGI) conditions. Our results showed that culturing 3T3-L1 adipocytes under HGI conditions increased triglyceride accumulation with concomitant decrease of ATGL and HSL expression, inducing insulin resistance in 3T3-L1 adipocytes. We also found that triglyceride accumulation was significantly inhibited and the levels of ATGL/HSL increased after melatonin or piromelatine treatment. The effects of melatonin/piromelatine (10 nM) were counteracted by pretreatment with the relatively selective MT2 receptor antagonist luzindole (100 nM). In this study, our data demonstrate that piromelatine reverses high glucose and high insulin-induced triglyceride accumulation in 3T3-L1 adipocytes, possibly through up-regulating of ATGL and HSL expression via a melatonin-dependent manner.

  14. The pharmacological effects of morroniside and loganin isolated from Liuweidihuang Wan, on MC3T3-E1 cells.

    Science.gov (United States)

    Li, Manyu; Wang, Wei; Wang, Ping; Yang, Kun; Sun, Hui; Wang, Xijun

    2010-10-21

    Liuweidihuang wan (LW), initially a well-known formula for curing "wu chi wu ruan", is commonly used nowadays for clinical treatment of postmenopausal osteoporosis (PO), but the identity of the effective substance(s) remains unclear. The present study was designed to evaluate the effects of morroniside and loganin isolated from LW on the proliferation, differentiation and apoptosis of MC3T3-E1 cells, as well as the possible mechanism of action. Morroniside and loganin had no effects on the proliferation of MC3T3-E1 cells, but both susbtances could improve the activity of alkaline phosphatase (ALP), and increase the contents of collagen type I and osteocalcin. Simultaneously, the mRNA expression of caspase-3, capase-9, RANKL was down-regulated and that of bcl-2 was up-regulated, which partially explains the anti-osteoporosis mechanism in MC3T3-E1 cells. In conclusion, morroniside and loganin may directly promote the differentiation and inhibit the apoptosis of MC3T3-E1 cells, and accordingly indirectly reduce bone resorption, which makes them promising natural drugs leads for treating PO in the near future.

  15. The Pharmacological Effects of Morroniside and Loganin Isolated from Liuweidihuang Wan, on MC3T3-E1 Cells

    Directory of Open Access Journals (Sweden)

    Xijun Wang

    2010-10-01

    Full Text Available Liuweidihuang Wan (LW, initially a well-known formula for curing “wu chi wu ruan”, is commonly used nowadays for clinical treatment of Postmenopausal Osteoporosis (PO, but the identity of the effective substance(s remains unclear. The present study was designed to evaluate the effects of morroniside and loganin isolated from LW on the proliferation, differentiation and apoptosis of MC3T3-E1 cells, as well as the possible mechanism of action. Morroniside and loganin had no effects on the proliferation of MC3T3-E1 cells, but both susbtances could improve the activity of alkaline phosphatase (ALP, and increase the contents of collagen type I and osteocalcin. Simultaneously, the mRNA expression of caspase-3, capase-9, RANKL was down-regulated and that of bcl-2 was up-regulated, which partially explains the anti-osteoporosis mechanism in MC3T3-E1 cells. In conclusion, morroniside and loganin may directly promote the differentiation and inhibit the apoptosis of MC3T3-E1 cells, and accordingly indirectly reduce bone resorption, which makes them promising natural drugs leads for treating PO in the near future.

  16. Casein kinase 2 regulates the active uptake of the organic osmolyte taurine in NIH3T3 mouse fibroblasts

    DEFF Research Database (Denmark)

    Jacobsen, Jack H; Clement, Christian A; Friis, Martin B;

    2008-01-01

    T to ER but has no detectable effect on TauT protein expression. On the other hand, CK2 inhibition increases the affinity of TauT towards Na(+ )and reduces the Na(+)/taurine stoichiometry for active taurine uptake. It is suggested that CK2 controls the cellular taurine uptake in unperturbated NIH3T3 cells...

  17. Molecular mechanism of 9-cis-retinoic acid inhibition of adipogenesis in 3T3-L1 cells

    Energy Technology Data Exchange (ETDEWEB)

    Sagara, Chiaki; Takahashi, Katsuhiko [Laboratory of Physiological Chemistry, Institute of Medicinal Chemistry, Hoshi University, Shinagawa, Tokyo 142-8501 (Japan); Kagechika, Hiroyuki [School of Biomedical Science, Institute of Biomaterials and Bioengineering, Tokyo Medical and Dental University, Chiyoda, Tokyo 101-0062 (Japan); Takahashi, Noriko, E-mail: t-noriko@hoshi.ac.jp [Laboratory of Physiological Chemistry, Institute of Medicinal Chemistry, Hoshi University, Shinagawa, Tokyo 142-8501 (Japan)

    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.

  18. Integrated FEM-DBEM simulation of crack propagation in AA2024-T3 FSW butt joints considering manufacturing effects

    DEFF Research Database (Denmark)

    Sonne, Mads Rostgaard; Carlone, P.; Citarella, R.

    2015-01-01

    of the welding process of precipitation hardenable AA2024-T3 aluminum alloy is employed to infer the process induced residual stress field. The reliability of the FEM simulations with respect to the induced residual stresses is assessed comparing numerical outcomes with experimental data obtained by means...

  19. Thoracoscopic sympathectomy of T2 and T3 ganglions for palmar hyperhydrosis

    Directory of Open Access Journals (Sweden)

    Toolabi K

    2008-05-01

    Full Text Available Background: Hyperhydrosis, excessive sweating, can profoundly affect the quality of life of the patient, with severe impairment of daily activities, social relationships and occupational activities. The purpose of this study was to evaluate the outcome of thoracoscopic sympathectomy in patients with palmar hyperhydrosis.Methods: In a clinical trial at Imam Khomeini, Milad and Velenjak Hospitals from 2003 to 2006, 33 patients older than five years of age with palmar hyperhydrosis underwent thoracoscopic sympathectomy of T2 and T3 ganglions. The variables regarding operation duration, length of hospital stay and early post-op complications were recorded. In addition, the quality of life of the patients was evaluated using the DLQI questionnaire, while the satisfaction of the patients was evaluated using the visual analogue scale before, immediately after and six months after the operation.Results: The mean age of the patients was 23 years and 63.6% of our patients were female. The sites of hyperhydrosis were hand, foot and armpit in 72.7% of the patients. In 90.91% of the cases, disease onset occurred during childhood and 78.79% of the patients had a family history of hyperhydrosis. Hyperhydrosis impaired function and had social, interpersonal and emotional effects in 72.7% of the cases. The mean duration of the operation was 40.3 minutes and the mean hospital stay was 1.45 days. The most common early post-op complications were atelectasis (7.57%, short-term pulmonary complications (3.03% and pleural effusion (3.03%. Compensatory hyperhydrosis occurred in 60% of the cases, while 18.75% experienced gustatory sweating. Palms were reported to be totally dry immediately after and six months after the operation in 81.82% of the cases, while 18.18% reported acceptable levels of perspiration. The mean level of satisfaction immediately after and six months after the procedure was 0.81. All patients felt improvement in all aspects of their lives

  20. Ghrelin inhibits the apoptosis of MC3T3-E1 cells through ERK and AKT signaling pathway

    Energy Technology Data Exchange (ETDEWEB)

    Liang, Qiu-Hua; Liu, Yuan; Wu, Shan-Shan; Cui, Rong-Rong; Yuan, Ling-Qing, E-mail: allenylq@hotmail.com; Liao, Er-Yuan, E-mail: eyliao@21cn.com

    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. Itm2a silencing rescues lamin A mediated inhibition of 3T3-L1 adipocyte differentiation.

    Science.gov (United States)

    Davies, Stephanie J; Ryan, James; O'Connor, Patrick B F; Kenny, Elaine; Morris, Derek; Baranov, Pavel V; O'Connor, Rosemary; McCarthy, Tommie V

    2017-08-18

    Dysregulation of adipose tissue metabolism is associated with multiple metabolic disorders. One such disease, known as Dunnigan-type familial partial lipodystrophy (FPLD2) is characterized by defective fat metabolism and storage. FPLD2 is caused by a specific subset of mutations in the LMNA gene. The mechanisms by which LMNA mutations lead to the adipose specific FPLD2 phenotype have yet to be determined in detail. We used RNA-Seq analysis to assess the effects of wild-type (WT) and mutant (R482W) lamin A on the expression profile of differentiating 3T3-L1 mouse preadipocytes and identified Itm2a as a gene that was upregulated at 36 h post differentiation induction in these cells. In this study we identify Itm2a as a novel modulator of adipogenesis and show that endogenous Itm2a expression is transiently downregulated during induction of 3T3-L1 differentiation. Itm2a overexpression was seen to moderately inhibit differentiation of 3T3-L1 preadipocytes while shRNA mediated knockdown of Itm2a significantly enhanced 3T3-L1 differentiation. Investigation of PPARγ levels indicate that this enhanced adipogenesis is mediated through the stabilization of the PPARγ protein at specific time points during differentiation. Finally, we demonstrate that Itm2a knockdown is sufficient to rescue the inhibitory effects of lamin A WT and R482W mutant overexpression on 3T3-L1 differentiation. This suggests that targeting of Itm2a or its related pathways, including autophagy, may have potential as a therapy for FPLD2.

  2. Effect of Ganoderma applanatum mycelium extract on the inhibition of adipogenesis in 3T3-L1 adipocytes.

    Science.gov (United States)

    Kim, Ji-Eun; Park, Sung-Jin; Yu, Mi-Hee; Lee, Sam-Pin

    2014-10-01

    Ganoderma applanatum (GA) and related fungal species have been used for over 2000 years in China to prevent and treat various human diseases. However, there is no critical research evaluating the functionality of GA grown using submerged culture technology. This study aimed to evaluate the effects of submerged culture GA mycelium (GAM) and its active components (protocatechualdehyde [PCA]) on preadipocyte differentiation of 3T3-L1 cells. Mouse-derived preadipocyte 3T3-L1 cells were treated with differentiation inducers in the presence or absence of GAM extracts. We determined triglyceride accumulations, glycerol-3-phosphate dehydrogenase (GPDH) activities, and differentiation makers. PCA, the active component of GAM extract, was also used to treat 3T3-L1 cells. The MTT assay showed that the GAM extract (0.01-1 mg/mL) was not toxic to 3T3-L1 preadipocyte. Treatment of cells with GAM extracts and its active components significantly decreased the GPDH activity and lipid accumulation, a marker of adipogenesis, in a dose-dependent manner. Western blot analysis results showed that the protein expression levels of peroxisome proliferator-activated receptor γ (PPARγ), CCAAT/enhancer-binding protein α (C/EBPα), and sterol regulatory element-binding protein 1 (SREBP1) were inhibited by the GAM extract. In addition, adipogenic-specific genes such as perilipin, fatty acid synthase (FAS), fatty acid transport protein 1 (FATP1), and fatty acid-binding protein 4 (FABP4) decreased in a dose-dependent manner. Quantitative high-performance liquid chromatography analysis showed that the GAM extract contained 1.14 mg/g PCA. GAM extracts suppressed differentiation of 3T3-L1 preadipocytes, in part, through altered regulation of PPARγ, C/EBPα, and SREBP1. These results suggest that GAM extracts and PCA may suppress adipogenesis by inhibiting differentiation of preadipocytes.

  3. Two Cases of Translocation t(3;6)(p14;p22): A Non Random Chromosomal Abnormality?

    Science.gov (United States)

    Lessard, M; Flandrin, G; Valensi, F; Gluckman, E

    1991-01-01

    It is generally accepted that the short arm of chromosome 6 is a likely site to be involved in chromosomal rearrangements of MDS/ANLL following radio/chemotherapy. We report here two cases of t(3;6)(p14;p22). One patient is a 55 years old male with a previous history of occupational exposure who developed, an acute megakaryoblastic leukemia after a preleukemic phase. Chromosome analysis showed a t(3;6)(p14;p22), associated with del (5)(q14q31), -7, with variations and a trend to hypoploidy. The second patient is a 33 years old man, with chronic myeloid leukemia treated with Hydroxyurea (HU), HU + $aL-IFN and $aL-IFN alone. The first cytogenetic study before treatment, showed a t(9;22)(q34;q11). In the following months the patient had simultaneously t(9;22)(q34;q11) + t(3;6)(p14;p22) in a minority and thereafter in all the mitoses, with progressive deterioration, megakaryocyte abnormalities, but no blast crisis. Our patients are compared with the only 5 other published cases with t(3;6)(p14;p22), who shared some common features, namely a past history of chemo/radiotherapy or exposure to chemical mutagens and an association with other, so-called "secondary" chromosome aberrations, on segments 3p, 5q, 7q, 12p and 17p. We suggest that this uncommon translocation t(3;6) is nonrandom. It is worth noting that band 6p21 is the site of pim 1 oncogen, and that a fragile site is located on band 3p14.

  4. Post-mastectomy radiotherapy in pT3N0M0 breast cancer: is it needed?

    Science.gov (United States)

    Helintö, M; Blomqvist, C; Heikkilä, P; Joensuu, H

    1999-09-01

    It is not been established whether breast cancer patients who have a primary tumor 5 cm or larger but no axillary nodal or distant metastases at the time of the diagnosis (pT3N0M0) benefit from post-operative radiation therapy after mastectomy. We identified 81 patients with T3N0M0 breast cancer out of the total of 4190 breast cancer patients treated in one university radiotherapy department from 1987 to 1994 from the department patient registry, and examined the clinical records and histopathological slides. Only 38 of the 81 patients had true pT3N0M0 breast cancer after the review (0.9% of the 4190 new breast cancer patients registered in the department from 1987 to 1994). Three (60%) of the five patients who were not treated with post-operative radiation therapy developed locoregional recurrence of breast cancer as compared with only three (9%) of the 33 patients who were given post-operative radiotherapy during a median follow-up of 58 months (P = 0.0003). Patients who were given post-operative radiotherapy had a better distant disease-free survival rate (P = 0.04) and overall survival rate (P = 0.03) than the ones who were not treated with radiation therapy after surgery. Of the 29 patients who had chest wall irradiation only, one had in-field recurrence at the surgical scar, one both at the scar and the unirradiated axilla, and only one (3%) solely in the axilla. Patients with true pT3N0M0 breast cancer are rare. The results suggest that women with pT3N0M0 breast cancer benefit from post-operative radiotherapy, but the value of irradiating the dissected ipsilateral axilla remains unsettled.

  5. Rhizobium-legume symbiosis in the absence of Nod factors: two possible scenarios with or without the T3SS.

    Science.gov (United States)

    Okazaki, Shin; Tittabutr, Panlada; Teulet, Albin; Thouin, Julien; Fardoux, Joël; Chaintreuil, Clémence; Gully, Djamel; Arrighi, Jean-François; Furuta, Noriyuki; Miwa, Hiroki; Yasuda, Michiko; Nouwen, Nico; Teaumroong, Neung; Giraud, Eric

    2016-01-01

    The occurrence of alternative Nod factor (NF)-independent symbiosis between legumes and rhizobia was first demonstrated in some Aeschynomene species that are nodulated by photosynthetic bradyrhizobia lacking the canonical nodABC genes. In this study, we revealed that a large diversity of non-photosynthetic bradyrhizobia, including B. elkanii, was also able to induce nodules on the NF-independent Aeschynomene species, A. indica. Using cytological analysis of the nodules and the nitrogenase enzyme activity as markers, a gradient in the symbiotic interaction between bradyrhizobial strains and A. indica could be distinguished. This ranged from strains that induced nodules that were only infected intercellularly to rhizobial strains that formed nodules in which the host cells were invaded intracellularly and that displayed a weak nitrogenase activity. In all non-photosynthetic bradyrhizobia, the type III secretion system (T3SS) appears required to trigger nodule organogenesis. In contrast, genome sequence analysis revealed that apart from a few exceptions, like the Bradyrhizobium ORS285 strain, photosynthetic bradyrhizobia strains lack a T3SS. Furthermore, analysis of the symbiotic properties of an ORS285 T3SS mutant revealed that the T3SS could have a positive or negative role for the interaction with NF-dependent Aeschynomene species, but that it is dispensable for the interaction with all NF-independent Aeschynomene species tested. Taken together, these data indicate that two NF-independent symbiotic processes are possible between legumes and rhizobia: one dependent on a T3SS and one using a so far unknown mechanism.

  6. Rhizobium–legume symbiosis in the absence of Nod factors: two possible scenarios with or without the T3SS

    Science.gov (United States)

    Okazaki, Shin; Tittabutr, Panlada; Teulet, Albin; Thouin, Julien; Fardoux, Joël; Chaintreuil, Clémence; Gully, Djamel; Arrighi, Jean- François; Furuta, Noriyuki; Miwa, Hiroki; Yasuda, Michiko; Nouwen, Nico; Teaumroong, Neung; Giraud, Eric

    2016-01-01

    The occurrence of alternative Nod factor (NF)-independent symbiosis between legumes and rhizobia was first demonstrated in some Aeschynomene species that are nodulated by photosynthetic bradyrhizobia lacking the canonical nodABC genes. In this study, we revealed that a large diversity of non-photosynthetic bradyrhizobia, including B. elkanii, was also able to induce nodules on the NF-independent Aeschynomene species, A. indica. Using cytological analysis of the nodules and the nitrogenase enzyme activity as markers, a gradient in the symbiotic interaction between bradyrhizobial strains and A. indica could be distinguished. This ranged from strains that induced nodules that were only infected intercellularly to rhizobial strains that formed nodules in which the host cells were invaded intracellularly and that displayed a weak nitrogenase activity. In all non-photosynthetic bradyrhizobia, the type III secretion system (T3SS) appears required to trigger nodule organogenesis. In contrast, genome sequence analysis revealed that apart from a few exceptions, like the Bradyrhizobium ORS285 strain, photosynthetic bradyrhizobia strains lack a T3SS. Furthermore, analysis of the symbiotic properties of an ORS285 T3SS mutant revealed that the T3SS could have a positive or negative role for the interaction with NF-dependent Aeschynomene species, but that it is dispensable for the interaction with all NF-independent Aeschynomene species tested. Taken together, these data indicate that two NF-independent symbiotic processes are possible between legumes and rhizobia: one dependent on a T3SS and one using a so far unknown mechanism. PMID:26161635

  7. Hh/Gli antagonist in acute myeloid leukemia with CBFA2T3-GLIS2 fusion gene

    Directory of Open Access Journals (Sweden)

    Riccardo Masetti

    2017-01-01

    Full Text Available Abstract Background CBFA2T3-GLIS2 is a fusion gene found in 17% of non-Down syndrome acute megakaryoblastic leukemia (non-DS AMKL, FAB M7 and in 8% of pediatric cytogenetically normal acute myeloid leukemia (CN-AML, in association with several French-American-British (FAB subtypes. Children with AML harboring this aberration have a poor outcome, regardless of the FAB subtype. This fusion gene drives a peculiar expression pattern and leads to overexpression of some of Hedgehog-related genes. GLI-similar protein 2 (GLIS2 is closely related to the GLI family, the final effectors of classic Hedgehog pathway. These observations lend compelling support to the application of GLI inhibitors in the treatment of AML with the aberration CBFA2T3-GLIS2. GANT61 is, nowadays, the most potent inhibitor of GLI family proteins. Methods We exposed to GANT61 AML cell lines and primary cells positive and negative for CBFA2T3-GLIS2 and analyzed the effect on cellular viability, induction of apoptosis, cell cycle, and expression profile. Results As compared to AML cells without GLIS2 fusion, GANT61 exposure resulted in higher sensitivity of both cell lines and primary AML cells carrying CBFA2T3-GLIS2 to undergo apoptosis and G1 cell cycle arrest. Remarkably, gene expression studies demonstrated downregulation of GLIS2-specific signature genes in both treated cell lines and primary cells, in comparison with untreated cells. Moreover, chromatin immunoprecipitation analysis revealed direct regulation by GLIS2 chimeric protein of DNMT1 and DNMT3B, two genes implicated in important epigenetic functions. Conclusions Our findings indicate that the GLI inhibitor GANT61 may be used to specifically target the CBFA2T3-GLIS2 fusion gene in pediatric AML.

  8. Expression of human insulin gene wrapped with chitosan nanoparticles in NIH3T3 cells and diabetic rats

    Institute of Scientific and Technical Information of China (English)

    Li NIU; Yan-cheng XU; Hai-ying XIE; Zhe DAI; Hui-qin TANG

    2008-01-01

    Aim: To study the expression of human insulin gene wrapped with chitosan nanoparticles in NIH3T3 cells and diabetic rats. Methods: pCMV.Ins, an expression plasmid of the human insulin gene, was constructed. In total, 100 μg pCMV.Ins wrapped with chitosan nanoparticles (chitosan-pCMV.Ins) was transfected to NIH3T3 cells and diabetes rats through lavage and coloclysis, respectively. The transfected cells were grown in Dulbecco's modified Eagle's medium, containing G418, for 72 h after transfection. The clones were selected and continued to grow in G418 medium for 24 d. The expression of human insulin was detected by immunohistochemistry. Human insulin in the culture medium of transfected cells was measured. Fasting blood glucose and plasma human insulin of diabetic rats were measured for 5 d after transfection. RT-PCR and Western blotting were performed to confirm the expression of the human insulin gene in diabetic rats. Results: Approximately 10% of NIH3T3 cells transfected by chitosan-pCMV.Ins expressed human insulin. Human insulin in the culture medium of NIH3T3 cells transfected by chitosan-pCMV.Ins significantly increased compared with that of the control group (P<0.01). Fasting blood glucose levels of the lavage group and the coloclysis group decreased significantly in 5 d (P<0.01) in comparison, while plasma insulin levels were much higher (P<0.01). The human insulin gene mRNA and human insulin were only detected in the lavage and the coloclysis groups. Conclusion: The human insulin gene can be transfected and expressed successfully by chitosan-pCMV.Ins in NIH3T3 cells and diabetes rats, which indicates that chitosan is a promising, non-viral vector for gene expression.

  9. Dlx2基因过表达与前成骨细胞系MC3T3-E1成骨分化过程中的细胞凋亡和周期调控%Effects of Dlx2 overexpression on cell cycle regulation and apoptosis of preosteoblast cells MC3T3-E1 during osteogenic differentiation process

    Institute of Scientific and Technical Information of China (English)

    孙昊; 王旭东; 代杰文; 卢境婷; 沈国芳

    2012-01-01

    背景:Dlx2 基因在颅神经嵴细胞迁徙进入第一鳃弓过程和颅颌面骨骼发育中起重要作用,但Dlx2 基因对成骨细胞分化过程中细胞凋亡和细胞周期调控的影响尚未见报道.目的:观察Dlx2 基因过表达对前成骨细胞系MC3T3-E1 成骨分化过程中细胞凋亡和细胞周期调控的影响.方法:构建反转录病毒pMSCV-puro-Dlx2 并转染矿化诱导液培养下的MC3T3-E1 细胞,构建稳定过表达Dlx2 基因的细胞系MC3T3-E1-Dlx2.RT-PCR 和Western blot 验证Dlx2 基因过表达细胞系的建立.Annexin V/PI 双染色后流式细胞分选检测细胞凋亡,PI/RNase 双染色后流式细胞分选检测细胞周期变化.结果与结论:实验成功构建稳定过表达Dlx2 基因的细胞系MC3T3-E1-Dlx2.发现Dlx2 基因过表达促进细胞凋亡(P < 0.05),同时阻滞细胞周期于G1/G0 期(P < 0.05),减低细胞增殖性,促进细胞分化行使功能.%BACKGROUND: Dlx2 plays an important role in the regulation of cranial neural crest cells migration and craniofacial skeleton development. However, the effects of D1×2 on apoptosis and cell cycle in osteoblast differentiation are not reported. OBJECTIVE: To investigate the effects of D1×2 overexpression on cell cycle regulation and apoptosis in MC3T3-E1 preosteoblastcells differentiation. METHODS: Anti-retroviral pMSCV-puro-Dlx2 was transfected into MC3T3-E1 cell s whi ch were cultured in mineralization induced fluid. MC3T3-E1-Dlx2 cell lines stably overexpressing Dlx2 were constructed. Cell lines of Dlx2 over-expression were determined by RT-PCR and western blot. Apoptosis was detected by flow cytometry after Annexin V/PI and PI/Rnase staining and cell cycle changes were detected by flow cytometry after PI/Rnase staining. RESULTS AND CONCLUSION: MC3T3-E1-D1×2 cell lines was successfully constructed. Cell cycle was blocked in G1/G0 phase (P < 0.05) and apoptosis was up-regulated (P < 0.05) in Dlx2 overexpression cell model. Dlx2 overexpression

  10. A Quantified Ginseng (Panax ginseng C.A. Meyer Extract Influences Lipid Acquisition and Increases Adiponectin Expression in 3T3-L1 Cells

    Directory of Open Access Journals (Sweden)

    Chia-Rou Yeo

    2011-01-01

    Full Text Available A Panax ginseng extract (PGE with a quantified amount of ginsenosides was utilized to investigate its potential to inhibit proliferation, influence lipid acquisition and adiponectin expression in 3T3-L1 cells. Seven fingerprint ginsenosides were quantified using high performance liquid chromatography and their respective molecular weights were further confirmed via LC-ESI-MS analysis from four different extraction methods. Extraction using methanol under reflux produced significantly higher amounts of ginsenosides. The methanol extract consisted of Rg1 (47.40 ± 4.28 mg/g, dry weight of extract, Re (61.62 ± 5.10 mg/g, Rf (6.14 ± 0.28 mg/g, Rb1 (21.73 ± 1.29 mg/g, Rc (78.79 ± 4.15 mg/g, Rb2 (56.80 ± 3.79 mg/g, Rd (5.90 ± 0.41 mg/g. MTT analysis showed that PGE had a concentrationdependent cytotoxic effect on 3T3-L1 preadipocyte and the LC50 value was calculated to be 18.2 ± 5 μg/mL. Cell cycle analysis showed minimal changes in all four phases. Differentiating adipocytes treated with ginseng extract had a visible decrease in lipid droplets formation measured by Oil red O staining. Consequently, triglycerides levels in media significantly (P < 0.05 decreased by 39.5% and 46.1% when treated at concentrations of 1 μg/mL and 10 μg/mL compared to untreated control cells. Western blot analysis showed that the adiponectin protein expression was significantly (P < 0.05 increased at 10 μg/mL, but not at 1 μg/mL. A quantified PGE reduced the growth of 3T3-L1 cells, down-regulated lipid accumulation and up-regulated adiponectin expression in the 3T3-L1 adipocyte cell model.

  11. The clinical experience of T3-4 thoracic sympathectomy in the treatment of hyperhidrosis%T3~4交感神经链切断术治疗手汗症的临床经验

    Institute of Scientific and Technical Information of China (English)

    徐彪; 吴君旭; 周晓; 沙纪名; 赵旭东; 石开虎

    2014-01-01

    目的 总结胸腔镜下T3~4交感神经链切断术治疗原发性手汗症的临床经验.方法 回顾性分析胸腔镜下T3~4交感神经链切断术治疗80例原发性手汗症患者的临床资料.结果 80例患者手术均获成功,术后手掌多汗症状立即消失,手术时间(42.5±15.7) min,住院时间(3.9±0.6)d.无霍纳综合征及死亡病例.62例患者随访6~24个月,发生轻度代偿性多汗26例,均无复发.结论 胸腔镜下T3~4交感神经链切断术是治疗原发性手汗症的一种安全、有效的微创治疗方法.%Objective To summarize the clinical experience of T3-4 thoracic sympathectomy in the treatment of primary hyperhidrosis.Methods The clinical data of 80 patients with primary hyperhidrosis who underwent T3-4 thoracic sympathectomy were analyzed retrospectively.Results The operation was successfully performed on all patients.The symptom of palmar hyperhidrosis vanished in all patients,the operative time was (42.5 ± 15.7) min,the length of stay in hospital was (3.9 ± 0.6) d.No death and Horner syndrome occurred.All the patients were followed up for 6-24 months,compensatory hyperhidrosis was found in 26 patients,and no recurrence was found.Conclusion T3-4 thoracic sympathectomy is a safe and effective minimally invasive treatment for primary hyperhidrosis.

  12. Amelioration of mitochondrial dysfunction-induced insulin resistance in differentiated 3T3-L1 adipocytes via inhibition of NF-κB pathways.

    Science.gov (United States)

    Bakar, Mohamad Hafizi Abu; Sarmidi, Mohamad Roji; Kai, Cheng Kian; Huri, Hasniza Zaman; Yaakob, Harisun

    2014-12-02

    A growing body of evidence suggests that activation of nuclear factor kappa B (NF-κB) signaling pathways is among the inflammatory mechanism involved in the development of insulin resistance and chronic low-grade inflammation in adipose tissues derived from obese animal and human subjects. Nevertheless, little is known about the roles of NF-κB pathways in regulating mitochondrial function of the adipose tissues. In the present study, we sought to investigate the direct effects of celastrol (potent NF-κB inhibitor) upon mitochondrial dysfunction-induced insulin resistance in 3T3-L1 adipocytes. Celastrol ameliorates mitochondrial dysfunction by altering mitochondrial fusion and fission in adipocytes. The levels of oxidative DNA damage, protein carbonylation and lipid peroxidation were down-regulated. Further, the morphology and quantification of intracellular lipid droplets revealed the decrease of intracellular lipid accumulation with reduced lipolysis. Moreover, massive production of the pro-inflammatory mediators tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) were markedly depleted. Insulin-stimulated glucose uptake activity was restored with the enhancement of insulin signaling pathways. This study signified that the treatments modulated towards knockdown of NF-κB transcription factor may counteract these metabolic insults exacerbated in our model of synergy between mitochondrial dysfunction and inflammation. These results demonstrate for the first time that NF-κB inhibition modulates mitochondrial dysfunction induced insulin resistance in 3T3-L1 adipocytes.

  13. Hop and Acacia Phytochemicals Decreased Lipotoxicity in 3T3-L1 Adipocytes, db/db Mice, and Individuals with Metabolic Syndrome

    Directory of Open Access Journals (Sweden)

    Deanna M. Minich

    2010-01-01

    Full Text Available The plant-based compounds rho-iso-alpha acids (RIAA from Humulus lupulus (hops and proanthocyanidins (PAC from Acacia nilotica have been shown to modulate insulin signaling in vitro. We investigated their effects on triglyceride (TG deposition in 3T3-L1 adipocytes, glucose and insulin in obese mouse models, and metabolic syndrome markers in adults with metabolic syndrome. The combination of RIAA and PAC synergistically increased TG content and adiponectin secretion in 3T3-L1 adipocytes under hyperinsulinemic conditions and reduced glucose or insulin in obese mice. In a clinical trial, tablets containing 100 mg RIAA and 500 mg PAC or placebo were administered to metabolic syndrome subjects (3 tablets/day, n=35; 6 tablets/day, n=34; or placebo, n=35 for 12 weeks. Compared to placebo, subjects taking 3 tablets daily showed greater reductions in TG, TG  :  HDL, fasting insulin, and HOMA scores. The combination of RIAA  :  PAC at 1  :  5 (wt  :  wt favorably modulates dysregulated lipids in insulin resistance and metabolic syndrome.

  14. Amelioration of Mitochondrial Dysfunction-Induced Insulin Resistance in Differentiated 3T3-L1 Adipocytes via Inhibition of NF-κB Pathways

    Directory of Open Access Journals (Sweden)

    Mohamad Hafizi Abu Bakar

    2014-12-01

    Full Text Available A growing body of evidence suggests that activation of nuclear factor kappa B (NF-κB signaling pathways is among the inflammatory mechanism involved in the development of insulin resistance and chronic low-grade inflammation in adipose tissues derived from obese animal and human subjects. Nevertheless, little is known about the roles of NF-κB pathways in regulating mitochondrial function of the adipose tissues. In the present study, we sought to investigate the direct effects of celastrol (potent NF-κB inhibitor upon mitochondrial dysfunction-induced insulin resistance in 3T3-L1 adipocytes. Celastrol ameliorates mitochondrial dysfunction by altering mitochondrial fusion and fission in adipocytes. The levels of oxidative DNA damage, protein carbonylation and lipid peroxidation were down-regulated. Further, the morphology and quantification of intracellular lipid droplets revealed the decrease of intracellular lipid accumulation with reduced lipolysis. Moreover, massive production of the pro-inflammatory mediators tumor necrosis factor-α (TNF-α and interleukin-1β (IL-1β were markedly depleted. Insulin-stimulated glucose uptake activity was restored with the enhancement of insulin signaling pathways. This study signified that the treatments modulated towards knockdown of NF-κB transcription factor may counteract these metabolic insults exacerbated in our model of synergy between mitochondrial dysfunction and inflammation. These results demonstrate for the first time that NF-κB inhibition modulates mitochondrial dysfunction induced insulin resistance in 3T3-L1 adipocytes.

  15. MicroRNA-24 promotes 3T3-L1 adipocyte differentiation by directly targeting the MAPK7 signaling

    Energy Technology Data Exchange (ETDEWEB)

    Jin, Min, E-mail: min_jin@zju.edu.cn [Division of Reproductive Medicine & Infertility, The Second Affiliated Hospital, School of Medicine, Zhejiang University, 88#, Jiefang Rd., Hangzhou, Zhejiang, 310009 (China); Wu, Yutao; Wang, Jing [School of Medicine, Zhejiang University, 288# Yuhangtang Rd, Hangzhou, Zhejiang, 310003 (China); Chen, Jian; Huang, Yiting; Rao, Jinpeng; Feng, Chun [Division of Reproductive Medicine & Infertility, The Second Affiliated Hospital, School of Medicine, Zhejiang University, 88#, Jiefang Rd., Hangzhou, Zhejiang, 310009 (China)

    2016-05-20

    Over the past years, MicroRNAs (miRNAs) act as a vital role in harmony with gene regulation and maintaining cellular homeostasis. It is well testified that miRNAshave been involved in numerous physiological and pathological processes, including embryogenesis, cell fate decision, and cellular differentiation. Adipogenesis is an organized process of cellular differentiation by which pre-adipocytes differentiate towards mature adipocytes, and it is tightly modulated by a series of transcription factors such as peroxisome proliferator-activated receptor γ (PPAR-γ) and sterol regulatory-element binding proteins 1 (SREBP1). However, the molecular mechanisms underlying the connection between miRNAs and adipogenesis-related transcription factors remain obscure. In this study, we unveiled that miR- 24 was remarkably upregulated during 3T3-L1 adipogenesis. Overexpression of miR-24 significantly promoted 3T3-L1 adipogenesis, as evidenced by its ability to increase the expression of PPAR-γ and SREBP1, lipid droplet formation and triglyceride (TG) accumulation. Furthermore, we found that neither ectopic expression of miR-24nor miR-24 inhibitor affect cell proliferation and cell cycle progression. Finally, we demonstrated that miR-24 plays the modulational role by directly repressing MAPK7, a key number in the MAPK signaling pathway. These data indicate that miR-24 is a novel positive regulator of adipocyte differentiation by targeting MAPK7, which provides new insights into the molecular mechanism of miRNA-mediated cellular differentiation. -- Highlights: •We firstly found miR-24 was upregulated in 3T3-L1 pre-adipocytes differentiation. •miR-24 promoted 3T3-L1 pre-adipocytes differentiation while silencing the expression of miR-24 had an opposite function. •miR-24 regulated 3T3-L1 differentiation by directly targeting MAPK7 signaling pathway. •miR-24did not affect 3T3-L1 pre-adipocytes cellular proliferation.

  16. Numerical analysis of static performance comparison of friction stir welded versus riveted 2024-T3 aluminum alloy stiffened panels

    Science.gov (United States)

    Shao, Qing; He, Yuting; Zhang, Teng; Wu, Liming

    2014-07-01

    Most researches on the static performance of stiffened panel joined by friction stir welding(FSW) mainly focus on the compression stability rather than shear stability. To evaluate the potential of FSW as a replacement for traditional rivet fastening for stiffened panel assembly in aviation application, finite element method(FEM) is applied to compare compression and shear stability performances of FSW stiffened panels with stability performances of riveted stiffened panels. FEMs of 2024-T3 aluminum alloy FSW and riveted stiffened panels are developed and nonlinear static analysis method is applied to obtain buckling pattern, buckling load and load carrying capability of each panel model. The accuracy of each FEM of FSW stiffened panel is evaluated by stability experiment of FSW stiffened panel specimens with identical geometry and boundary condition and the accuracy of each FEM of riveted stiffened panel is evaluated by semi-empirical calculation formulas. It is found that FEMs without considering weld-induced initial imperfections notably overestimate the static strengths of FSW stiffened panels. FEM results show that, buckling patterns of both FSW and riveted compression stiffened panels represent local buckling of plate between stiffeners. The initial buckling waves of FSW stiffened panel emerge uniformly in each plate between stiffeners while those of riveted panel mainly emerge in the mid-plate. Buckling patterns of both FSW and riveted shear stiffened panels represent local buckling of plate close to the loading corner. FEM results indicate that, shear buckling of FSW stiffened panel is less sensitive to the initial imperfections than compression buckling. Load carrying capability of FSW stiffened panel is less sensitive to the initial imperfections than initial buckling. It can be concluded that buckling loads of FSW panels are a bit lower than those of riveted panels whereas carrying capabilities of FSW panels are almost equivalent to those of riveted

  17. 胸腔镜下T3胸交感神经切断术治疗手汗症62例临床分析%Thoracoscopic T3 Sympathectomy for Palmar Hyperhidrosis: Analysis on 62 Cases

    Institute of Scientific and Technical Information of China (English)

    钟卫权; 吕坤; 廖金文; 廖承辉

    2012-01-01

    Objective To discuss and analyze our clinical experience on thoracoscopic T, sympathectomy for palmar hyperhidrosis. Methods From July 2008 to March 2011, we performed thoracoscopic T3 sympathectomy on totally 62 patients with palmar hyperhidrosis. With incisions made at the 5th intercostal space on the midaxillary line, and 3rd intercostal space on the anterior axillary line as observation and operation holes, respectively, we cut the thoracic sympathetic chain and communicating branches. Results The procedure was completed successfully on all the 62 patients. The mean operation time for bilateral procedure was 24. 3 minutes (19-36 minutes) , and mean hospital stay was 4. 5 days (3-6 days) . Nine patients developed postoperative pleural effusion but were all cured afterwards. Totally 56 patients received follow-up for 3-12 months with a mean of 7. 2 months, during which, 2 patients showed recurrent mild palmar hyperhidrosis and 3 patients had mild compensatory sweating. Conclusion Thoracoscopic T3 sympathectomy is an effective, minimally invasive and safe therapy for palmar hyperhidrosis with a low recurrence rate.%目的 探讨胸腔镜下T3胸交感神经切断术治疗手汗症的疗效. 方法 2008年7月~2011年3月对62例手汗症行胸腔镜下经操作孔电凝钩切断T3胸交感神经及可能存在的交通支. 结果 62例手术均获成功,手术时间(双侧)19~36 min,平均24.3 min.住院3~6d,平均4.5d.术后出现胸腔积液9例,4例行胸腔穿刺抽液,5例经保守治疗治愈.56例术后随访3~12个月,平均7.2月,2例复发轻度手汗,3例出现轻度代偿性出汗. 结论 胸腔镜下T3胸交感神经切断术治疗手汗症疗效确切,不易复发,术后代偿性多汗发生率低,安全性高.

  18. HUBUNGAN KANDUNGAN KLOR SERUM DENGAN HORMON T3/T4 PADA ANAK SEKOLAH DI DAERAH GONDOK ENDEMIK

    Directory of Open Access Journals (Sweden)

    Sukati Saidin

    2012-11-01

    Full Text Available ASSOCIATION OF SERUM CHLOR CONTENT WITH T3/T4 HORMONEIN SCHOOL CHILDREN IN IODINE DEFICIENCY REGION.Background: The National Mapping Survey of IDD (1998 found that 7% of sub districts In Indonesia was regarded as severe endemic goitre area (TGR>30%. The high TGR prevalence, beside as a result of low iodized salt consumption (< 30 ppm, It was assumed as the effect of exposure of goitrogenic agent such as chlorine. Based on observation in Karawang sub district showed people had food habit to consume fish contaminated by insecticide used for killing milk fish predator or salted fish which had also contaminated by insecticide used during process of fish drying. Insecticide raw material consists of chlorine which can not be broken by heat or oxidation. Previous study by Gaitan E. (1986 found that chlorine component could inhibit iodine metabolism to form mono and di-iodotyrosine as precursor of T3 and T4 hormones.Objectives: The aim of this study was to find an association of serum chlorine as a reflection of chlorine consumption from daily food with T3 and T4 hormone.Methods: Research design was case control. Study was conducted in Karawang district, West Java. The subject were elementary school children in the fourth, fifth and sixth grades with positive goitre at grade I and II by palpation. Sample size was 140 children divided into two groups, case group (70 children and control group (70 children. Main data collected was chlorine consumption from daily food, serum chlorine, serum T3 and T4 hormones as well as anthropometries.Results: The result showed that chlorine consumption from food was relatively greater in case group (135.9 ugr/day than in control group (129.9 ug/day but statistically it was not significant. Serum chlorine content in case group (1 14.8 mmol/L was significantly higher than in control group (102.1 mmol/L. Serum T4 hormone in case group (7.3 ug/dl was significantly lower than in control group (9.5 ug/dl. Serum T3 hormone in

  19. Magnetic Levitation of MC3T3 Osteoblast Cells as a Ground-Based Simulation of Microgravity

    Science.gov (United States)

    Kidder, Louis S.; Williams, Philip C.; Xu, Wayne Wenzhong

    2009-01-01

    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

  20. p53-independent upregulation of p21WAF1 in NIH 3T3 cells malignantly transformed by mot-2

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    ot-2 protein is shown to interact with p53 and inhibit its transcriptional activation function.Mot-2 overexpressing stable clones of NIH 3T3 cells were malignantly transformed,however,they had a high level of expression of a p53 downstream gene,P21waf1.The present study was undertaken to elucidate possible molecular mechanism(s) of such upregulation.An increased level of P21waf1 expression was detected in stable transfectants although an exogenous reporter gene driven by P21waf1 promoter exhibited lower activity in these cells suggesting that some post-transcriptional mechanism contributes to upregulation.Western analyses of transient and stable clones revealed that upregulation of P21waf1 in stable NIH 3T3/mot-2 cells may be mediated by cyclin D1 and cdk-2.

  1. 适用于T3工况空调的蒸发器优化设计

    Institute of Scientific and Technical Information of China (English)

    宋培刚

    2012-01-01

    Through air conditioner simulation and test verification, this article will describe the way to increase 6.9% capacity and 5.9% efficiency by optimizing T1 air conditioner evaporator flow distribution within 0 cost increase. This research indicates that adverse flow designed evaporator can also be applied to T3 air conditioners%本文分别通过空调仿真和实验验证,实现了在空调成本不变的情况下,优化T1工况空调蒸发器的流程布置,将空调能力提升6.9%,能效提高5.9%。本文研究表明采用逆流的形式蒸发器,可以很好的适用于T3工况空调。

  2. PLA2 - a major regulator of volume-sensitive taurine release in NIH3T3 fibroblasts

    DEFF Research Database (Denmark)

    Lambert, I. H.

    2006-01-01

    Release of the organic osmolyte taurine efflux from NIH3T3 cells is increased by osmotic cell swelling in hypotonic medium and decreased by osmotic cell shrinkage in hypertonic medium. Release of arachidonic acid is increased under hypotonic conditions if oxidation of the fatty acid via the 5...... and taurine in NIH3T3 cells under isotonic conditions. The potentiating effect of melittin on arachidonic release and taurine efflux is substantially increased in osmotically swollen cells and almost abolished in osmotic shrunken cells. H2O2 potentiates the melittin-induced taurine efflux under isotonic...... conditions but has only a minor effect on the melittin-induced taurine efflux under hypertonic conditions. Bromoenol lactone and manoalide, known inhibitors of Ca2+-independent phospholipase A2 (iPLA2) and secretory phospholipase A2 (sPLA2), respectively, reduce arachidonic acid and taurine release from NIH3...

  3. Germline polymorphisms may act as predictors of response to preoperative chemoradiation in locally advanced T3 rectal tumors

    DEFF Research Database (Denmark)

    Spindler, Karen-Lise G; Nielsen, Jens N; Lindebjerg, Jan;

    2007-01-01

    with locally advanced T3 rectal tumors were analyzed for thymidylate synthase, epidermal growth factor receptor Sp1-216, and epidermal growth factor A61G gene polymorphisms by polymerase chain reaction. Treatment consisted of preoperative radiotherapy (total dose 65 Gy) and concomitant chemotherapy (Uftoral......PURPOSE: Patients with locally advanced T3 rectal tumors who present with complete pathologic response to preoperative chemoradiation have a low rate of local recurrence and an excellent prognosis. Predictive markers for complete pathologic response are needed with the perspective of improving...... individualized treatment of these patients. This study was designed to investigate the predictive value of a new combination of three gene polymorphisms: thymidylate synthase, epidermal growth factor receptor Sp1-216, and epidermal growth factor A61G. METHODS: Pretreatment blood samples from 60 patients...

  4. HuB localizes to polysomes and alters C/EBP-beta expression in 3T3-L1 adipocytes.

    Science.gov (United States)

    Gantt, Kira R; Jain, Renu G; Dudek, Ronald W; Pekala, Phillip H

    2004-01-16

    The RNA binding protein HuB was ectopically expressed in 3T3-L1 preadipocytes and localized primarily to the nucleus, as the cells differentiated HuB redistributed to the cytosol and on analysis localized to the dense polysomes. Electron micrographs confirm association of HuB with the ribosomes in the adipocytes consistent with a proposed role in control of translation and mRNA stability. Examination of the expression of C/EBP-beta in the cells expressing HuB relative to the parental 3T3-L1 adipocytes demonstrated an alteration in the LAP to LIP ratio. The data support a role for endogenous Hu proteins in the differentiation process, potentially affecting the rate of differentiation by controlling the concentration of the dominant negative transcription inhibitor, LIP.

  5. Enhancing T3 and cAMP responsive gene participation in the thermogenic regulation of fuel oxidation pathways

    OpenAIRE

    2010-01-01

    OBJECTIVE: We sought to identify glycolysis, glycogenolysis, lipolysis, Krebs cycle, respiratory chain, and oxidative phosphorylation enzymes simultaneously regulated by T3 and cAMP. MATERIALS AND METHODS: We performed in silico analysis of 56 promoters to search for cis-cAMP (CREB) and cis-thyroid (TRE) response elements, considering UCP1, SERCA2 and glyceraldehyde 3-phosphate dehydrogenase as reference. Only regulatory regions with prior in vitro validation were selected. RESULTS: 29/56 enz...

  6. 13-Methylberberine, a berberine analogue with stronger anti-adipogenic effects on mouse 3T3-L1 cells

    OpenAIRE

    Chow, Yit-Lai; Sogame, Mami; Sato, Fumihiko

    2016-01-01

    Lipid metabolism modulation is a main focus of metabolic syndrome research, an area in which many natural and synthetic chemicals are constantly being screened for in vitro and in vivo activity. Berberine, a benzylisoquinoline plant alkaloid, has been extensively investigated for its anti-obesity effects and as a potential cholesterol and triglyceride-lowering drug. We screened 11 protoberberine and 2 benzophenanthridine alkaloids for their anti-adipogenic effects on 3T3-L1 adipocytes and fou...

  7. Functional expression of 5-HT{sub 2A} receptor in osteoblastic MC3T3-E1 cells

    Energy Technology Data Exchange (ETDEWEB)

    Hirai, Takao; Kaneshige, Kota; Kurosaki, Teruko [Department of Molecular Pharmacology, Faculty of Pharmacy and Pharmaceutical Sciences, Fukuyama University, 1 Gakuen-cho, Fukuyama, Hiroshima 729-0292 (Japan); Nishio, Hiroaki, E-mail: nishio@fupharm.fukuyama-u.ac.jp [Department of Molecular Pharmacology, Faculty of Pharmacy and Pharmaceutical Sciences, Fukuyama University, 1 Gakuen-cho, Fukuyama, Hiroshima 729-0292 (Japan)

    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. Effects of C-reactive protein on adipokines genes expression in 3T3-L1 adipocytes

    Energy Technology Data Exchange (ETDEWEB)

    Yuan, Guoyue, E-mail: yuanguoyue@hotmail.com [Department of Endocrinology, The Affiliated Hospital of Jiangsu University, Zhenjiang, Jiangsu 212001 (China); Jia, Jue; Di, Liangliang [Department of Endocrinology, The Affiliated Hospital of Jiangsu University, Zhenjiang, Jiangsu 212001 (China); Zhou, Libin [Ruijin Hospital, Center of Molecular Medicine, Shanghai Institute of Endocrine and Metabolic Diseases, State Key Laboratory of Medical Genomics, Shanghai Jiaotong University Medical School, 197, Ruijin Road II, Shanghai 200025 (China); Dong, Sijing; Ye, Jingjing; Wang, Dong; Yang, Ling; Wang, Jifang [Department of Endocrinology, The Affiliated Hospital of Jiangsu University, Zhenjiang, Jiangsu 212001 (China); Li, Lianxi [Department of Endocrinology and Metabolism, Shanghai Jiao Tong University Affiliated Sixth People' s Hospital, 600, Yishan Road, Shanghai 200233 (China); Yang, Ying [Ruijin Hospital, Center of Molecular Medicine, Shanghai Institute of Endocrine and Metabolic Diseases, State Key Laboratory of Medical Genomics, Shanghai Jiaotong University Medical School, 197, Ruijin Road II, Shanghai 200025 (China); Mao, Chaoming [Department of Endocrinology, The Affiliated Hospital of Jiangsu University, Zhenjiang, Jiangsu 212001 (China); Chen, Mingdao, E-mail: mingdaochensh@yahoo.com [Ruijin Hospital, Center of Molecular Medicine, Shanghai Institute of Endocrine and Metabolic Diseases, State Key Laboratory of Medical Genomics, Shanghai Jiaotong University Medical School, 197, Ruijin Road II, Shanghai 200025 (China)

    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.

  9. Citrus aurantium flavonoids inhibit adipogenesis through the Akt signaling pathway in 3T3-L1 cells

    Directory of Open Access Journals (Sweden)

    Kim Gon-Sup

    2012-04-01

    Full Text Available Abstract Background Obesity is a health hazard that is associated with a number of diseases and metabolic abnormalities, such as type-2 diabetes, hypertension, dyslipidemia, and coronary heart disease. In the current study, we investigated the effects of Citrus aurantium flavonoids (CAF on the inhibition of adipogenesis and adipocyte differentiation in 3T3-L1 cells. Methods During adipocyte differentiation, 3T3-L1 cells were treated with 0, 10, and 50 μg/ml CAF, and then the mRNA and protein expression of adipogenesis-related genes was assayed. We examined the effect of CAF on level of phosphorylated Akt in 3T3-L1 cells treated with CAF at various concentrations during adipocyte differentiation. Results The insulin-induced expression of C/EBPβ and PPARγ mRNA and protein were significantly down-regulated in a dose-dependent manner following CAF treatment. CAF also dramatically decreased the expression of C/EBPα, which is essential for the acquisition of insulin sensitivity by adipocytes. Moreover, the expression of the aP2 and FAS genes, which are involved in lipid metabolism, decreased dramatically upon treatment with CAF. Interestingly, CAF diminished the insulin-stimulated serine phosphorylation of Akt (Ser473 and GSK3β (Ser9, which may reduce glucose uptake in response to insulin and lipid accumulation. Furthermore, CAF not only inhibited triglyceride accumulation during adipogenesis but also contributed to the lipolysis of adipocytes. Conclusions In the present study, we demonstrate that CAF suppressed adipogenesis in 3T3-L1 adipocytes. Our results indicated that CAF down-regulates the expression of C/EBPβ and subsequently inhibits the activation of PPARγ and C/EBPα. The anti-adipogenic activity of CAF was mediated by the inhibition of Akt activation and GSK3β phosphorylation, which induced the down-regulation of lipid accumulation and lipid metabolizing genes, ultimately inhibiting adipocyte differentiation.

  10. Insertion of the T3 DNA polymerase thioredoxin binding domain enhances the processivity and fidelity of Taq DNA polymerase

    OpenAIRE

    2003-01-01

    Insertion of the T3 DNA polymerase thioredoxin binding domain (TBD) into the distantly related thermostable Taq DNA polymerase at an analogous position in the thumb domain, converts the Taq DNA polymerase from a low processive to a highly processive enzyme. Processivity is dependent on the presence of thioredoxin. The enhancement in processivity is 20–50-fold when compared with the wild-type Taq DNA polymerase or to the recombinant polymerase in the absence of thioredoxin. The recombinant Taq...

  11. Inhibition of adipogenesis and leptin production in 3T3-L1 adipocytes by a derivative of meridianin C

    Energy Technology Data Exchange (ETDEWEB)

    Park, Yu-Kyoung [Department of Molecular Medicine, College of Medicine, Keimyung University, 1095 Dalgubeoldaero, Dalseo-gu, Daegu 704-701 (Korea, Republic of); Lee, Tae-Yoon [Department of Microbiology, College of Medicine, Yeungnam University, 170 Hyunchung-Ro, Nam-gu, Daegu 705-717 (Korea, Republic of); Choi, Jong-Soon [Division of Life Science, Korea Basic Science Institute, 169-148 Gwahakro, Yuseong-gu, Daejeon 305-333 (Korea, Republic of); Hong, Victor Sukbong [Department of Chemistry, College of Natural Sciences, Keimyung University, 1095 Dalgubeoldaero, Dalseo-gu, Daegu 704-701 (Korea, Republic of); Lee, Jinho, E-mail: jinho@gw.kmu.ac.kr [Department of Chemistry, College of Natural Sciences, Keimyung University, 1095 Dalgubeoldaero, Dalseo-gu, Daegu 704-701 (Korea, Republic of); Park, Jong-Wook, E-mail: j303nih@dsmc.or.kr [Department of Immunology, College of Medicine, Keimyung University, 1095 Dalgubeoldaero, Dalseo-gu, Daegu 704-701 (Korea, Republic of); Jang, Byeong-Churl, E-mail: jangbc123@gw.kmu.ac.kr [Department of Molecular Medicine, College of Medicine, Keimyung University, 1095 Dalgubeoldaero, Dalseo-gu, Daegu 704-701 (Korea, Republic of)

    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.

  12. Caracterización de la respuesta a fractura de las aleaciones de aluminio 2024-O y 2024-T3

    OpenAIRE

    Monslave, A.; Morales, R.

    2004-01-01

    The fracture behavior of 2024-0 (annealed) and 2024-T3 (precipitation hardened) aluminum alloys used in aeronautical applications have been characterized. The study of the annealed alloy was carried out through the concept of Essential Specific Work of Fracture, using the ESIS protocol. DENT (Double Edge Notch Tension) samples were used, varying the ligament length in order to determine the relationship between the essential specific work of fracture and the thickness of the material. In the ...

  13. Mammalian ste20-like kinase and SAV1 promote 3T3-L1 adipocyte differentiation by activation of PPARγ.

    Directory of Open Access Journals (Sweden)

    Byoung Hee Park

    Full Text Available The mammalian ste20 kinase (MST signaling pathway plays an important role in the regulation of apoptosis and cell cycle control. We sought to understand the role of MST2 kinase and Salvador homolog 1 (SAV1, a scaffolding protein that functions in the MST pathway, in adipocyte differentiation. MST2 and MST1 stimulated the binding of SAV1 to peroxisome proliferator-activated receptor γ (PPARγ, a transcription factor that plays a key role in adipogenesis. The interaction of endogenous SAV1 and PPARγ was detected in differentiating 3T3-L1 adipocytes. This binding required the kinase activity of MST2 and was mediated by the WW domains of SAV1 and the PPYY motif of PPARγ. Overexpression of MST2 and SAV1 increased PPARγ levels by stabilizing the protein, and the knockdown of SAV1 resulted in a decrease of endogenous PPARγ protein in 3T3-L1 adipocytes. During the differentiation of 3T3-L1 cells into adipocytes, MST2 and SAV1 expression began to increase at 2 days when PPARγ expression also begins to increase. MST2 and SAV1 significantly increased PPARγ transactivation, and SAV1 was shown to be required for the activation of PPARγ by rosiglitazone. Finally, differentiation of 3T3-L1 cells was augmented by MST2 and SAV1 expression and inhibited by knockdown of MST1/2 or SAV1. These results suggest that PPARγ activation by the MST signaling pathway may be a novel regulatory mechanism of adipogenesis.

  14. Inhibitory effect of apocynin on methylglyoxal-mediated glycation in osteoblastic MC3T3-E1 cells.

    Science.gov (United States)

    Suh, Kwang Sik; Rhee, Sang Youl; Kim, Young Seol; Choi, Eun Mi

    2015-04-01

    Methylglyoxal (MG), a highly reactive metabolite of hyperglycemia, can enhance protein glycation, oxidative stress or inflammation. The present study investigated the effects of apocynin on the mechanisms associated with MG toxicity in osteoblastic MC3T3-E1 cells. Pretreatment of MC3T3-E1 cells with apocynin prevented the MG-induced protein glycation and formation of intracellular reactive oxygen species and mitochondrial superoxide in MC3T3-E1 cells. In addition, apocynin increased glutathione levels and restored the activity of glyoxalase I inhibited by MG. These findings suggest that apocynin provide a protective action against MG-induced cell damage by reducing oxidative stress and by increasing the MG detoxification system. Apocynin treatment decreased the levels of proinflammatory cytokines such as tumor necrosis factor-α and interleukin-6 induced by MG. Additionally, the nitric oxide level reduced by MG was significantly increased by apocynin. These findings indicate that apocynin might exert its therapeutic effects via upregulation of glyoxalase system and antioxidant activity. Taken together, apocynin may prove to be an effective treatment for diabetic osteopathy.

  15. Effects of Berberine on Adipose Tissues and Kidney Function in 3T3-L1 Cells and Spontaneously Hypertensive Rats.

    Science.gov (United States)

    Kishimoto, Aya; Dong, Shi-Fen; Negishi, Hiroko; Yasui, Naomi; Sun, Jian-Ning; Ikeda, Katsumi

    2015-09-01

    We aimed to investigate the effect of berberine on adipose tissues, as well as its effect on renal injury in 3T3-L1 cells and spontaneously hypertensive rats. 3T3-L1 cells were cultured and treated with berberine (5-20 pM) from days 3 to 8. Berberine added to the cultured medium could significantly down-regulate transcription factors, including CCAAT/enhancer binding protein β, CCAAT/enhancer binding protein a, and peroxisome pro liferator-activated receptor y, and suppress peroxisome proliferator-activated receptor target genes, such as adipocyte fatty acid binding protein and fatty acid synthase, and inhibit 3T3-Ll fibroblast differentiation to adipocytes. Male spontaneously hypertensive rats received either 150 mg/day of berberine or saline orally for 8 weeks. Compared with the control, berberine-treated rats exhibited significant reductions in body weight gain (p Berberine-treated rats significantly decreased urinary albumin excretion, a marker of renal injury (p berberine decreased the adipose tissues weight and attenuated renal injury in spontaneously hypertensive rats. Based on these results, berberine has an important role in regulating adipose tissues. These results suggest the protective effect of berberine on metabolic syndrome related diseases, such as renal injury.

  16. The influence of staphylococcin T (StT) on the mouse fibroblasts (3T3) viability, an in vitro experiment.

    Science.gov (United States)

    Drewa, Tomasz; Krawczyk, Agnieszka; Gospodarek, Eugenia; Jachna-Sawicka, Katarzyna; Bugalski, Roman; Kierzenkowska-Mila, Celestyna

    2006-01-01

    Staphylococcin T (StT) is a bacteriocin, which can serve as an antibiotic. The influence of StT on the mammalian cells viability should be examined before its possible applications. The aim of this work was to study the influence of StT on mouse fibroblasts viability in vitro. StT was delivered as a deposit in agar. 3T3 fibroblasts were used in viability tests. Cells were incubated with StT for 24 and 48 h. In the first controls, cells grew without the agar block, in the second controls, the agar block contained no StT. Trypan blue exclusion test was performed. Growth inhibition of sensitive strain SAU-209P was observed after 24 h incubation with cell culture medium containing StT. Antimicrobial activity of StT in culture medium was confirmed even after 2 months. The viability of 3T3 cells in the second control group was 50% lower than in the first and 25% higher than in StT treated group during the first day. It was noticed mechanical damage of growing cells caused by agar blocks. The number of cells cultured with StT was similar to that of second control group after 2 days. There were no changes in morphology of cells in all groups. It follows that StT had unimportant influence on the fibroblasts (3T3) viability in culture. StT may be considered for future animal trails.

  17. WSF-P-1, a novel AMPK activator, promotes adiponectin multimerization in 3T3-L1 adipocytes.

    Science.gov (United States)

    Wang, Yao; Zhang, Yudian; Wang, Yunyun; Peng, Han; Rui, Jian; Zhang, Zhijie; Wang, Shifa; Li, Zhen

    2017-08-01

    Adiponectin, an adipokine with insulin-sensitizing effect, is secreted from adipocytes into circulation as high, medium, and low molecular weight forms (HMW, MMW, and LMW). The HMW adiponectin oligomers possess the most potent insulin-sensitizing activity. WSF-P-1(N-methyl-1,2,3,4,5,6-hexahydro-1,1,5,5-tetramethyl-7H-2,4α-methanonaphthalen-7-amine) is derived from natural sesquiterpene longifolene by chemical modifications. We found that WSF-P-1 activates AMPK in both 3T3-L1 adipocytes and 293T cells in this study. Activation of AMPK by WSF-P-1 promotes the assembly of HMW adiponectin and increases the HMW/total ratio of adiponectin in 3T3-L1 adipocytes. We demonstrated that the Ca(2+)-dependent CaMKK signaling pathway is involved in WSF-P-1-induced AMPK activation and adiponectin multimerization. WSF-P-1 also activates GLUT1-mediated glucose uptake in 3T3-L1 adipocytes, making it a potential drug candidate for the treatment of type 2 diabetes, obesity, and other obesity-related metabolic diseases.

  18. Differential response of MC3T3-E1 and human mesenchymal stem cells to inositol hexakisphosphate.

    Science.gov (United States)

    Arriero, María Del Mar; Ramis, Joana M; Perelló, Joan; Monjo, Marta

    2012-01-01

    Inositol hexakisphosphate (IP6) has been found to have an important role in biomineralization. Because the complete mechanism of action of IP6 on osteoblasts is not fully understood and its potential use in the primary prevention of osteoporosis, we examined the direct effect of IP6 on cell viability and differentiation of MC3T3-E1 cells and on differentiation of human umbilical cord mesenchymal stem cells (hUC-MSCs). We show that IP6 has different effects depending on the origin of the cell target. Thus, while IP6 decreased gene expression of osteoblast markers and mineralization in MC3T3-E1 cells without negatively affecting cell viability and ALP activity, an increase in gene expression of ALP was observed in hUC-MSCs committed to the osteoblastic lineage. This increasing effect of IP6 on ALP mRNA expression levels was reversed by the addition of a selective inhibitor of IP6 kinase, suggesting that the effect of IP6 might be due through its pyrophosphorylated derivatives. Besides, Rankl mRNA levels were decreased after IP6 treatment in MC3T3-E1 cells, pointing to a paracrine effect on osteoclasts. Our results indicate that IP6 has different effects on osteoblast differentiation depending on the cell type and origin. However, further studies are needed to examine the net effect of IP6 on bone formation and its potential as novel antiosteoporosis drug. Copyright © 2012 S. Karger AG, Basel.

  19. Prediction of forming limit curve (FLC) for Al-Li alloy 2198-T3 sheet using different yield functions

    Institute of Scientific and Technical Information of China (English)

    Li Xiaoqiang; Song Nan; Guo Guiqiang; Sun Zhonggang

    2013-01-01

    The Forming Limit Curve (FLC) of the third generation aluminum-lithium (Al-Li) alloy 2198-T3 is measured by conducting a hemispherical dome test with specimens of different widths. The theoretical prediction of the FLC of 2198-T3 is based on the M-K theory utilizing respectively the von Mises, Hill’48, Hosford and Barlat 89 yield functions, and the different predicted curves due to different yield functions are compared with the experimentally measured FLC of 2198-T3. The results show that though there are differences among the four predicted curves, yet they all agree well with the experimentally measured curve. In the area near the planar strain state, the predicted curves and experimentally measured curve are very close. The predicted curve based on the Hosford yield function is more accurate under the tension-compression strain states described in the left part of the FLC, while the accuracy is better for the predicted curve based on Hill’48 yield function under the tension-tension strain states shown in the right part.

  20. Chlamydia pneumoniae CopD translocator protein plays a critical role in type III secretion (T3S and infection.

    Directory of Open Access Journals (Sweden)

    David C Bulir

    Full Text Available Pathogenic Gram-negative bacteria use type III secretion (T3S to inject effector proteins into the host cell to create appropriate conditions for infection and intracellular replication. Chlamydia spp. are believed to use T3S to infect their host cell, and the translocator proteins are an essential component of this system. Chlamydia pneumoniae contains genes encoding two sets of translocator proteins; CopB and CopD, and CopB2 and CopD2. In this study, we identified novel interactions between CopD and three type III secretion proteins; namely, CopN, CdsN, and CdsF. We identified a CopD putative chaperone binding motif, PxLxxP, within the N-terminal region (CopD amino acids 120-125, which was necessary for interaction with its putative chaperone LcrH_1. Using size exclusion chromatography, we showed that CopD and LcrH_1 formed higher order structures in solution with CopD and LcrH_1 binding in a ratio of 1∶1, which is unique for T3SS translocator proteins. Lastly, we showed that antibodies to CopD reduced C. pneumoniae infectivity by >95%. Collectively, this data suggests that CopD plays a critical role in pathogenesis and likely functions as a hydrophobic translocator of the type III secretion system in Chlamydia pneumoniae.

  1. Phorbol esters enhance attachment of NIH/3T3 cells to laminin and type IV collagen substrates

    Energy Technology Data Exchange (ETDEWEB)

    Kato, Shigemi; Ben, T.L.; De Luca, L.M. (National Institutes of Health, Bethesda, MD (USA))

    1988-11-01

    The effect of phorbol esters on the adhesive properties of NIH/3T3 mouse fibroblasts was investigated using plastic substrates precoated with the extracellular matrix proteins fibronectin, collagen, and laminin. Treatment with phorbol 12-myristate 13-acetate (PMA) enhanced NIH/3T3 cell attachment to laminin and type IV collagen substrates but had little or no effect on attachment to fibronectin and type I collagen substrates. The effect of PMA in enhancing cell attachment to laminin and type IV collagen substrates was dose dependent between 10{sup {minus}9} and 10{sup {minus}7} M. PMA was effective as early as 30 min; the effect reached a maximum at 2 h and decreased gradually. Phorbol 12, 13-dibenzoate and phorbol 12, 13-diacetate were effective but to a lesser extent and phorbol 12-myristate and phorbol 13-acetate showed little or no effect. These results suggest that PMA may enhance NIH/3T3 cell adhesion through effects on laminin and type IV collagen receptors. Retinoic acid, which itself requires at least 6 h to show an effect on attachment, did not have any effect on cell attachment in 2 h and, if anything, slightly inhibited PMA-enhanced cell attachment to laminin and type IV collagen substrates.

  2. Inhibitory effects of green tea catechin on the lipid accumulation in 3T3-L1 adipocytes.

    Science.gov (United States)

    Lee, Mak-Soon; Kim, Chong-Tai; Kim, In-Hwan; Kim, Yangha

    2009-08-01

    The aim of the present study was to evaluate the effects of green tea (-)-epigallocatechin-3-gallate (EGCG) on the depletion of accumulated fat in differentiated 3T3-L1 adipocytes. Intracellular lipid accumulation was decreased significantly after 24 h of incubation with 10 microm EGCG, while the viability of adipocytes was reported to be unaffected. Under the same experimental conditions, the amount of glycerol released from cells into the medium was increased by 10 microm EGCG. The level of mRNA in the 3T3-L1 adipocytes was analysed by quantitative real-time RT-PCR. EGCG notably increased the mRNA level of hormone sensitive lipase (HSL), which catalyses the rate-limiting stage in hydrolysis of stored triacylglycerol to monoacylglycerol and free fatty acids. In conclusion, the experiment produced results which showed that green tea EGCG effectively depleted fat accumulation via the stimulation of lipolysis and increased HSL gene expression in 3T3-L1 adipocytes. These results may relate to the mechanism by which EGCG modulates lipolysis in adipocytes. Copyright 2009 John Wiley & Sons, Ltd.

  3. On the possibility of generating a 4-neutron resonance with a {\\boldmath $T=3/2$} isospin 3-neutron force

    CERN Document Server

    Hiyama, E; Carbonell, J; Kamimura, M

    2016-01-01

    We consider the theoretical possibility to generate a narrow resonance in the four neutron system as suggested by a recent experimental result. To that end, a phenomenological $T=3/2$ three neutron force is introduced, in addition to a realistic $NN$ interaction. We inquire what should be the strength of the $3n$ force in order to generate such a resonance. The reliability of the three-neutron force in the $T=3/2$ channel is exmined, by analyzing its consistency with the low-lying $T=1$ states of $^4$H, $^4$He and $^4$Li and the $^3{\\rm H} + n$ scattering. The {\\it ab initio} solution of the $4n$ Schr\\"{o}dinger equation is obtained using the complex scaling method with boundary conditions appropiate to the four-body resonances. We find that in order to generate narrow $4n$ resonant states a remarkably attractive $3N$ force in the $T=3/2$ channel is required.

  4. Aculeatin, a coumarin derived from Toddalia asiatica (L.) Lam., enhances differentiation and lipolysis of 3T3-L1 adipocytes

    Energy Technology Data Exchange (ETDEWEB)

    Watanabe, Akio, E-mail: watanabea@jfrl.or.jp [Japan Food Research Laboratories, Osaka 567-0085 (Japan); Food and Biodynamic Chemistry Laboratory, Graduate School of Agricultural Science, Tohoku University, Miyagi 981-8555 (Japan); Kato, Tsuyoshi; Ito, Yusuke; Yoshida, Izumi; Harada, Teppei; Mishima, Takashi; Fujita, Kazuhiro; Watai, Masatoshi [Japan Food Research Laboratories, Osaka 567-0085 (Japan); Nakagawa, Kiyotaka; Miyazawa, Teruo [Food and Biodynamic Chemistry Laboratory, Graduate School of Agricultural Science, Tohoku University, Miyagi 981-8555 (Japan)

    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.

  5. Enhanced effects of guggulsterone plus 1,25(OH)2D3 on 3T3-L1 adipocytes.

    Science.gov (United States)

    Rayalam, Srujana; Della-Fera, Mary Anne; Ambati, Suresh; Boyan, Barbara; Baile, Clifton A

    2007-12-21

    Guggulsterone (GS) and 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] have been shown to influence adipogenesis in 3T3-L1 cells. We investigated the ability of GS and 1,25(OH)2D3, alone and in combination to inhibit adipogenesis and induce apoptosis in 3T3-L1 adipocytes. Maturing preadipocytes were treated with 1,25(OH)2D3 in combination with GS for 6 days during differentiation. GS and 1,25(OH)2D3 each inhibited lipid accumulation, but the combination potentiated the inhibition of lipid accumulation. Apoptosis was increased by 1,25(OH)2D3 while GS had no effect, but GS + 1,25(OH)2D3 increased apoptosis more than either compound alone. Furthermore, GS + 1,25(OH)2D3 caused a potentiated decrease in the expression of aP2 and farnesoid X receptor expression more than either compound alone. In addition, 1,25(OH)2D3 increased vitamin D receptor expression after 6 days, while GS had no effect. GS + 1,25(OH)2D3, however, caused a potentiated increase in the expression of VDR. These findings show that GS potentiates 1,25(OH)2D3's anti-adipogenic and pro-apoptotic effects in maturing 3T3-L1 preadipocytes.

  6. Proinflammatory cytokine production and insulin sensitivity regulated by overexpression of resistin in 3T3-L1 adipocytes

    Directory of Open Access Journals (Sweden)

    Garvey W Timothy

    2006-07-01

    Full Text Available Abstract Resistin is secreted from adipocytes, and high circulating levels have been associated with obesity and insulin resistance. To investigate whether resistin could exert autocrine effects in adipocytes, we expressed resistin gene in 3T3-L1 fibroblasts using a lentiviral vector, and selected several stably-transduced cell lines under blasticidin selection. We observed that 3T3-L1 adipocytes expressing resistin have a decreased gene expression for related transcriptional factors (CCAAT/enhancer binding protein α(C/EBPα , peroxisome proliferator-activated receptor gamma (PPARγ, and adipocyte lipid binding protein (ALBP/aP2 which is one of target genes for the PPARγ during adipocyte differentiation,. Overexpression of resistin increased the levels of three proinflammatory cytokines, tumor necrosis factor alpha (TNFα, interleukin 6 (IL-6 and monocyte chemoattractant protein-1 (MCP-1, which play important roles for insulin resistance, glucose and lipid metabolisms during adipogenesis. Furthermore, overexpressing resistin in adipocytes inhibits glucose transport 4 (GLUT4 activity and its gene expression, reducing insulin's ability for glucose uptake by 30 %. In conclusion, resistin overexpression in stably transduced 3T3-L1 cells resulted in: 1 Attenuation of programmed gene expression responsible for adipogenesis; 2 Increase in expression of proinflammatory cytokines; 3 Decrease in insulin responsiveness of the glucose transport system. These data suggest a new role for resistin as an autocrine/paracrine factor affecting inflammation and insulin sensitivity in adipose tissue.

  7. Soy pinitol acts partly as an insulin sensitizer or insulin mediator in 3T3-L1 preadipocytes.

    Science.gov (United States)

    Do, Gyeong-Min; Choi, Myung-Sook; Kim, Hye-Jin; Woo, Myung-Nam; Lee, Mi-Kyung; Jeon, Seon-Min

    2008-02-01

    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/EBPalpha and PPARgamma 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/EBPalpha and PPARgamma in 3T3-L1 preadipocytes.

  8. Rapamycin/sodium hyaluronate binding on nano-hydroxyapatite coated titanium surface improves MC3T3-E1 osteogenesis

    Science.gov (United States)

    Liu, Chao; Dong, Jian Yong; Yue, Lin Lin; Liu, Shao Hua; Wan, Yi; Liu, Hong; Tan, Wan Ye; Guo, Qian Qian; Zhang, Dong

    2017-01-01

    Endosseous titanium (Ti) implant failure due to poor biocompatibility of implant surface remains a major problem for osseointegration. Improving the topography of Ti surface may enhance osseointegration, however, the mechanism remains unknown. To investigate the effect of modified Ti surface on osteogenesis, we loaded rapamycin (RA) onto nano-hydroxyapatite (HAp) coated Ti surface which was acid-etched, alkali-heated and HAp coated sequentially. Sodium hyaluronate (SH) was employed as an intermediate layer for the load of RA, and a steady release rate of RA was maintained. Cell vitality of MC3T3-E1 was assessed by MTT. Osteogenesis of MC3T3-E1 on this modified Ti surface was evaluated by alkaline phosphatase (ALP) activity, mineralization and related osteogenesis genes osteocalcin (OCN), osteopontin (OPN), Collagen-I and Runx2. The result revealed that RA/SH-loaded nano-HAp Ti surface was innocent for cell vitality and even more beneficial for cell osteogenesis in vitro. Furthermore, osteogenesis of MC3T3-E1 showed significant association with the mammalian target of rapamycin (mTOR) phosphorylation by RA, which required further study about the mechanism. The approach to this modified Ti surface presented in this paper has high research value for the development of Ti-based implant. PMID:28182765

  9. Long-term outcomes of surgery alone versus surgery following preoperative chemoradiotherapy for early T3 rectal cancer

    Science.gov (United States)

    Cho, Seung Hyun; Choi, Gyu-Seog; Kim, Gab Chul; Seo, An Na; Kim, Hye Jung; Kim, Won Hwa; Shin, Kyung-Min; Lee, So Mi; Ryeom, Hunkyu; Kim, See Hyung

    2017-01-01

    Abstract Recently, a few studies have raised the question of whether preoperative chemoradiotherapy (PCRT) is essential for all T3 rectal cancers. This case-matched study aimed to compare the long-term outcomes of surgery alone with those of PCRT + surgery for magnetic resonance imaging (MRI)-assessed T3ab (extramural depth of invasion ≤5 mm) and absent mesorectal fascia invasion (clear MRF) in mid/lower rectal cancer patients. From January 2006 to November 2012, 203 patients who underwent curative surgery alone (n = 118) or PCRT + surgery (n = 85) were enrolled in this retrospective study. A 1:1 propensity score-matched analysis was performed to eliminate the inherent bias. Case-matching covariates included age, sex, body mass index, histologic grade, carcinoembryonic antigen, operation method, follow-up period, tumor height, and status of lymph node metastasis. The end-points were the 5-year local recurrence (LR) rate and disease-free-survival (DFS). After propensity score matching, 140 patients in 70 pairs were included. Neither the 5-year LR rate nor the DFS was significantly different between the 2 groups (the 5-year LR rate, P = 0.93; the 5-year DFS, P = 0.94). The 5-year LR rate of the surgery alone was 2% (95% confidence interval [CI] 0.2%–10.9%) versus 2% (95% CI 0.2%–10.1%) in the PCRT + surgery group. The 5-year DFS of the surgery alone was 87% (95% CI 74.6%–93.7%) versus 88% (95% CI 77.8%–93.9%) in the PCRT + surgery group. In patients with MRI-assessed T3ab and clear MRF mid/lower rectal cancer, the long-term outcomes of surgery alone were comparable with those of the PCRT + surgery. The suggested MRI-assessed T3ab and clear MRF can be used as a highly selective indication of surgery alone in mid/lower T3 rectal cancer. Additionally, in those patients, surgery alone can be tailored to the clinical situation. PMID:28328820

  10. The Differentiation-and Proliferation-Inhibitory Effects of Sporamin from Sweet Potato in 3T3-L1 Preadipocytes

    Institute of Scientific and Technical Information of China (English)

    XIONG Zhi-dong; LI Peng-gao; MU Tai-hua

    2009-01-01

    The aim of this study was to investigate the effect of different concentrations of sporamin on the differentiation and proliferation of 3T3-L1 preadipocytes,providing the theoretical basis for the development of food to treat obesity and diabetes.The isolation and purification of sporamin from sweet potato species 55-2 were performed by ammonium sulphate precipitation in combination with ion-exchange and gel filtration chromatography.With berberine as a positive control,different concentrations of sporamin (0.000,0.125,0.025,0.250,0.500,and 1.000 mg mL-1) were used to treat 3T3-L1 preadipocytes.Intracellular fat accumulation and the degree of adipogenesis were quantified using Oil Red O staining and colorimetry.Preadipocytes differentiation was measured by 3-(4,5-dimethylthiazolyl-2-yl)-2,5-diphenyltetrazolium bromide (MTT)spectrophotometric assay.Two sporamin proteins,which were separated into sporamin A (31 kD) and sporamin B (22 kD),could be purified by ion-exchange and gel filtration chromatography.After being treated by different concentrations of sporamin,the differentiation of 3T3-L1 preadipocytes was significantly inhibited,compared with the positive control.When the sporamin solution concentration was 0.500 mg mL-1,the accumulation of lipid droplets within the cells was significantly decreased and the optical density (OD) value of the solution from destained Oil Red O reached to 0.35,which was the lowest value (P < 0.05).The proliferation of 3T3-L1 preadipocytes was significantly inhibited by treating at higher sporamin concentrations.In addition,the inhibitory effect was more obvious with the prolonged treatment time (P< 0.05).The differentiation and proliferation of 3T3-L1 preadipocytes could be inhibited significantly by the addition of higher concentration sporamin.It was,therefore,suggested that the sporamin was potentially effective for weight loss.

  11. Modest hypoxia significantly reduces triglyceride content and lipid droplet size in 3T3-L1 adipocytes

    Energy Technology Data Exchange (ETDEWEB)

    Hashimoto, Takeshi, E-mail: thashimo@fc.ritsumei.ac.jp [Faculty of Sport and Health Science, Ritsumeikan University, 1-1-1 Nojihigashi, Kusatsu, Shiga 525-8577 (Japan); Yokokawa, Takumi; Endo, Yuriko [Faculty of Sport and Health Science, Ritsumeikan University, 1-1-1 Nojihigashi, Kusatsu, Shiga 525-8577 (Japan); Iwanaka, Nobumasa [Ritsumeikan Global Innovation Research Organization, Ritsumeikan University, 1-1-1 Nojihigashi, Kusatsu, Shiga 525-8577 (Japan); Higashida, Kazuhiko [Faculty of Sport and Health Science, Ritsumeikan University, 1-1-1 Nojihigashi, Kusatsu, Shiga 525-8577 (Japan); Faculty of Sport Science, Waseda University, 2-579-15 Mikajima, Tokorozawa, Saitama 359-1192 (Japan); Taguchi, Sadayoshi [Faculty of Sport and Health Science, Ritsumeikan University, 1-1-1 Nojihigashi, Kusatsu, Shiga 525-8577 (Japan)

    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

  12. Roughness threshold for cell attachment and proliferation on plasma micro-nanotextured polymeric surfaces: the case of primary human skin fibroblasts and mouse immortalized 3T3 fibroblasts

    Science.gov (United States)

    Bourkoula, A.; Constantoudis, V.; Kontziampasis, D.; Petrou, P. S.; Kakabakos, S. E.; Tserepi, A.; Gogolides, E.

    2016-08-01

    Poly(methyl methacrylate) surfaces have been micro-nanotextured in oxygen plasmas with increasing ion energy, leading to micro-nanotopography characterized by increased root mean square roughness, correlation length and fractal dimension. Primary human skin fibroblasts and mouse immortalized 3T3 fibroblasts were cultured on these surfaces and the number of adhering cells, their proliferation rate and morphology (cytoplasm and nucleus area) were evaluated as a function of roughness height, correlation length, and fractal dimension. A roughness threshold behavior was observed for both types of cells leading to dramatic cell number decrease above this threshold, which is almost similar for the two types of cells, despite their differences in size and stiffness. The results are discussed based on two theoretical models, which are reconciled and unified when the elastic moduli and the size of the cells are taken into account.

  13. 催产素对3T3-L1脂肪细胞糖脂代谢的影响%Effect of oxytocin on glucose and lipid metabolism of 3T3-L1 adipocytes

    Institute of Scientific and Technical Information of China (English)

    朱天一; 钱唯韵; 汤冰倩; 胡浩; 俞淑琴; 孙文君; 袁国跃

    2014-01-01

    目的:观察催产素对3T3-L1脂肪细胞糖脂代谢的影响。方法3T3-L1前脂肪细胞体外培养,并诱导其分化成熟为脂肪细胞。研究催产素对脂肪细胞葡萄糖消耗量以及三酰甘油、游离脂肪酸和甘油的影响。采用实时荧光定量PCR法检测糖脂代谢相关基因GLUT-1、GLUT-4、ATGT、HSL的mRNA表达。结果与对照组比较,催产素20、50、100μg/mL组葡萄糖消耗量有所增加,且表现出剂量相关。催产素组较对照组的三酰甘油降低,而甘油和游离脂肪酸增高。催产素50μg/mL组中脂代谢相关基因HSL表达明显高于对照组,糖代谢相关基因GLUT-4 mRNA表达水平增加。结论催产素处理可减少3T3-L1细胞脂质合成、增加脂质分解作用,并可明显改善脂质积聚。%Objective To study the effect of oxytocin on glucose and lipid metabolism in 3T3-L1 adipocytes. Methods Preadipocytes from 3T3-L1 cell line were cultured in vitro and induced to differentiate to adipocytes. Mature adipocytes were treated with oxytocin. Glucose consumption, triglyceride, free fat acid, and glycerol levels were determined. The mRNA expression of differentiation marker genes such as GLUT-1, GLUT-4, ATGT, and HSL were evaluated by RT-PCR method. Results The glucose consumption in the oxytocin (20, 50, and 100μg/mL) groups were increased with dose-dependent relationship compared with control group. Triglyceride in the oxytocin group was lower than that in control group, while glycerol and free fatty acid decreased. There was significant increase of expression levels of lipid metabolism related gene HSL and sugar metabolism related gene GLUT-4 mRNA in the oxytocin (50μg/mL) group compared with control group. Conclusion Treatment of oxytocin may reduce 3T3-L1 cell lipid synthesis, increase lipid decomposition, and obviously improve lipid accumulation.

  14. A methodology for estimating seasonal cycles of atmospheric CO2 resulting from terrestrial net ecosystem exchange (NEE fluxes using the Transcom T3L2 pulse-response functions

    Directory of Open Access Journals (Sweden)

    K. R. Gurney

    2012-09-01

    Full Text Available We present a method for translating modeled terrestrial net ecosystem exchange (NEE fluxes of carbon into the corresponding seasonal cycles in atmospheric CO2. The method is based on the pulse-response functions from the Transcom 3 Level 2 (T3L2 atmospheric tracer transport model (ATM intercomparison. The new pulse-response method is considerably faster than a full forward ATM simulation, allowing CO2 seasonal cycles to be computed in seconds, rather than the days or weeks required for a forward simulation. Further, the results provide an estimate of the range of transport uncertainty across 13 different ATMs associated with the translation of surface NEE fluxes into an atmospheric signal. We evaluate the method against the results of archived forward ATM simulations from T3L2. The latter are also used to estimate the uncertainties associated with oceanic and fossil fuel influences. We present a regional breakdown at selected monitoring sites of the contribution to the atmospheric CO2 cycle from the 11 different T3L2 land regions. A test case of the pulse-response code, forced by NEE fluxes from the Community Land Model, suggests that for many terrestrial models, discrepancies between model results and observed atmospheric CO2 cycles will be large enough to clearly transcend ATM uncertainties.

  15. Clinical Effect of Low Dose Tetraiodothyronine in Hepatic Fibrosis Activity on Compensated Cirrhosis with T3 Degraded%小剂量甲状腺素对低T_3代偿期肝硬化的影响

    Institute of Scientific and Technical Information of China (English)

    陈洁

    2009-01-01

    目的 探讨代偿期肝硬化甲状腺激素水平的变化及小剂量甲状腺素对低T_3代偿期肝硬化患者肝纤维化的影响.方法 60名伴有T_3降低的代偿期肝硬化患者,随机分为两组,给予相同的保肝、营养支持治疗,治疗组另加左旋甲状腺素片25 mg/d,连续1个月,观察两组临床疗效、肝功能及肝纤维化指标的变化.结果 治疗组同对照组临床疗效无差异,治疗组血清AST、ALT、血清透明质酸(HA)、层粘连蛋白(LN)、Ⅲ型前胶原肽(PC-Ⅲ)、Ⅳ型胶原(C-Ⅳ)水平低于对照组,ALB无变化,A/G比值高于对照组.结论 外源性甲状腺素对T_3代偿期肝硬化患者能减轻肝纤维化的发展.%Objectives To investigate change of Tetraiodothyronine(TH)in serum on compensated cirrhosis with T_3 degraded and observe the effect of TH in hepatic fibrosis activity.Methods Sixty patients were randomly divided into experimental group and control group.The patients in experimental group and control group received same treatment,including protecting hepatic function and nutritional support.TH were taken to experimental group with 25 mg/d for a month.clinical effect were estimated,Hepatic function and hepatic fibrosis activity were measured in serum on all patients when the treatment was over.Results In comparison with control group,the levels of AST、ALT、HA、LN、PC-Ⅲand C-Ⅳin serum were lower,the level of ALB Was indiscriminate,and proportionality of A/G was higher in cirrhosis.Clinical effect was same in two groups.Conclusion The level of TH in serum was decreasing in compensation cirrhosis,and additional TH can decrease hepatic fibrosis activity,but can not improve clinical effect in short.

  16. Porting a 3D-model for the transport of reactive air pollutants to the parallel machine T3D

    NARCIS (Netherlands)

    Kessler, C.; Blom, J.G.; Verwer, J.G.

    1995-01-01

    Air pollution forecasting puts a high demand on the memory and the floating point performance of modern computers. For this kind of problems massively parallel computers are very promising, although the software tools and the I/O facilities on those machines are still under-developed. This report de

  17. Catabolism of Branched Chain Amino Acids Contributes Significantly to Synthesis of Odd-Chain and Even-Chain Fatty Acids in 3T3-L1 Adipocytes.

    Directory of Open Access Journals (Sweden)

    Scott B Crown

    Full Text Available The branched chain amino acids (BCAA valine, leucine and isoleucine have been implicated in a number of diseases including obesity, insulin resistance, and type 2 diabetes mellitus, although the mechanisms are still poorly understood. Adipose tissue plays an important role in BCAA homeostasis by actively metabolizing circulating BCAA. In this work, we have investigated the link between BCAA catabolism and fatty acid synthesis in 3T3-L1 adipocytes using parallel 13C-labeling experiments, mass spectrometry and model-based isotopomer data analysis. Specifically, we performed parallel labeling experiments with four fully 13C-labeled tracers, [U-13C]valine, [U-13C]leucine, [U-13C]isoleucine and [U-13C]glutamine. We measured mass isotopomer distributions of fatty acids and intracellular metabolites by GC-MS and analyzed the data using the isotopomer spectral analysis (ISA framework. We demonstrate that 3T3-L1 adipocytes accumulate significant amounts of even chain length (C14:0, C16:0 and C18:0 and odd chain length (C15:0 and C17:0 fatty acids under standard cell culture conditions. Using a novel GC-MS method, we demonstrate that propionyl-CoA acts as the primer on fatty acid synthase for the production of odd chain fatty acids. BCAA contributed significantly to the production of all fatty acids. Leucine and isoleucine contributed at least 25% to lipogenic acetyl-CoA pool, and valine and isoleucine contributed 100% to lipogenic propionyl-CoA pool. Our results further suggest that low activity of methylmalonyl-CoA mutase and mass action kinetics of propionyl-CoA on fatty acid synthase result in high rates of odd chain fatty acid synthesis in 3T3-L1 cells. Overall, this work provides important new insights into the connection between BCAA catabolism and fatty acid synthesis in adipocytes and underscores the high capacity of adipocytes for metabolizing BCAA.

  18. Catabolism of Branched Chain Amino Acids Contributes Significantly to Synthesis of Odd-Chain and Even-Chain Fatty Acids in 3T3-L1 Adipocytes.

    Science.gov (United States)

    Crown, Scott B; Marze, Nicholas; Antoniewicz, Maciek R

    2015-01-01

    The branched chain amino acids (BCAA) valine, leucine and isoleucine have been implicated in a number of diseases including obesity, insulin resistance, and type 2 diabetes mellitus, although the mechanisms are still poorly understood. Adipose tissue plays an important role in BCAA homeostasis by actively metabolizing circulating BCAA. In this work, we have investigated the link between BCAA catabolism and fatty acid synthesis in 3T3-L1 adipocytes using parallel 13C-labeling experiments, mass spectrometry and model-based isotopomer data analysis. Specifically, we performed parallel labeling experiments with four fully 13C-labeled tracers, [U-13C]valine, [U-13C]leucine, [U-13C]isoleucine and [U-13C]glutamine. We measured mass isotopomer distributions of fatty acids and intracellular metabolites by GC-MS and analyzed the data using the isotopomer spectral analysis (ISA) framework. We demonstrate that 3T3-L1 adipocytes accumulate significant amounts of even chain length (C14:0, C16:0 and C18:0) and odd chain length (C15:0 and C17:0) fatty acids under standard cell culture conditions. Using a novel GC-MS method, we demonstrate that propionyl-CoA acts as the primer on fatty acid synthase for the production of odd chain fatty acids. BCAA contributed significantly to the production of all fatty acids. Leucine and isoleucine contributed at least 25% to lipogenic acetyl-CoA pool, and valine and isoleucine contributed 100% to lipogenic propionyl-CoA pool. Our results further suggest that low activity of methylmalonyl-CoA mutase and mass action kinetics of propionyl-CoA on fatty acid synthase result in high rates of odd chain fatty acid synthesis in 3T3-L1 cells. Overall, this work provides important new insights into the connection between BCAA catabolism and fatty acid synthesis in adipocytes and underscores the high capacity of adipocytes for metabolizing BCAA.

  19. About lamination upper and convexification lower bounds on the free energy of monoclinic shape memory alloys in the context of T 3-configurations and R-phase formation

    Science.gov (United States)

    Fechte-Heinen, R.; Schlömerkemper, A.

    2016-11-01

    This work is concerned with different estimates of the quasiconvexification of multi-well energy landscapes of NiTi shape memory alloys, which models the overall behavior of the material. Within the setting of the geometrically linear theory of elasticity, we consider a formula of the quasiconvexification which involves the so-called energy of mixing.We are interested in lower and upper bounds on the energy of mixing in order to get a better understanding of the quasiconvexification. The lower bound on the energy of mixing is obtained by convexification; it is also called Sachs or Reuß lower bound. The upper bound on the energy of mixing is based on second-order lamination. In particular, we are interested in the difference between the lower and upper bounds. Our numerical simulations show that the difference is in fact of the order of 1% and less in martensitic NiTi, even though both bounds on the energy of mixing were rather expected to differ more significantly. Hence, in various circumstances it may be justified to simply work with the convexification of the multi-well energy, which is relatively easy to deal with, or with the lamination upper bound, which always corresponds to a physically realistic microstructure, as an estimate of the quasiconvexification. In order to obtain a potentially large difference between upper and lower bound, we consider the bounds along paths in strain space which involve incompatible strains. In monoclinic shape memory alloys, three-tuples of pairwise incompatible strains play a special role since they form so-called T 3-configurations, originally discussed in a stress-free setting. In this work, we therefore consider in particular numerical simulations along paths in strain space which are related to these T 3-configurations. Interestingly, we observe that the second-order lamination upper bound along such paths is related to the geometry of the T 3-configurations. In addition to the purely martensitic regime, we also consider

  20. Blister Test for Measurements of Adhesion and Adhesion Degradation of Organic Polymers on AA2024-T3

    Science.gov (United States)

    Rincon Troconis, Brendy Carolina

    A key parameter for the performance of corrosion protective coatings applied to metals is adhesion. Surface preparation prior to coating application is known to be critical, but there is a lack of understanding of what controls adhesion. Numerous techniques have been developed in the last decades to measure the adhesion strength of coatings to metals. Nonetheless, they are generally non-quantitative, non-reproducible, performed in dry conditions, or overestimate adhesion. In this study, a quantitative and reproducible technique, the Blister Test (BT), is used. The BT offers the ability to study the effects of a range of parameters, including the presence or absence of a wetting liquid, and simulates the stress situation in the coating/substrate interface. The effects of roughness and surface topography were studied by the BT and Optical Profilometry, using AA2024-T3 substrates coated with polyvinyl butyral (PVB). Random abrasion generated a surface with lower average roughness than aligned abrasion due to the continual cross abrasion of the grooves. The BT could discern the effects of different mechanical treatments. An adhesion strength indicator was defined and found to be a useful parameter. The effectiveness of standard adhesion techniques such as ASTM D4541 (Pull-off Test) and ASTM D3359 (Tape Test) was compared to the BT. Also, different attempts to measure adhesion and adhesion degradation of organic polymers to AA2024-T3 were tested. The pull-off test does not produce adhesive failure across the entire interface, while the tape test is a very qualitative technique and does not discern between the effects of different coating systems on the adhesion performance. The BT produces adhesive failure of the primer studied, is very reproducible, and is able to rank different coating systems. Therefore, it was found to be superior to the others. The approaches tested for adhesion degradation were not aggressive enough to have a measurable effect. The effects of

  1. Characterization of hypersensitive resistance to bacterial spot race T3 (Xanthomonas perforans) from tomato accession PI 128216.

    Science.gov (United States)

    Robbins, Matthew D; Darrigues, Audrey; Sim, Sung-Chur; Masud, Mohammed Abu Taher; Francis, David M

    2009-09-01

    Bacterial spot of tomato is caused by four species of Xanthomonas. The accession PI 128216 (Solanum pimpinellifolium) displays a hypersensitive reaction (HR) to race T3 strains (predominantely Xanthomonas perforans). We developed an inbred backcross (IBC) population (BC(2)S(5), 178 families) derived from PI 128216 and OH88119 (S. lycopersicum) as the susceptible recurrent parent for simultaneous introgression and genetic analysis of the HR response. These IBC families were evaluated in the greenhouse for HR to race T3 strain Xcv761. The IBC population was genotyped with molecular markers distributed throughout the genome in order to identify candidate loci conferring resistance. We treated the IBC population as a hypothesis forming generation to guide validation in subsequent crosses. Nonparametric analysis identified an association between HR and markers clustered on chromosome 11 (P P > 0.002). Further analysis of the IBC population suggested that markers on chromosome 6 and 11 failed to assort independently, a phenomenon known as gametic phase disequilibrium. Therefore, to validate marker-trait linkages, resistant IBC plants were crossed with OH88119 and BC(3)F(2) progeny were evaluated for HR in the greenhouse. In these subsequent populations, the HR response was associated with the chromosome 11 markers (P 0.25). Independent F(2) families were developed by crossing resistant IBC lines to OH8245, OH88119, and OH7530. These populations were genotyped, organized into classes based on chromosome 11 markers, and evaluated for resistance in the field. The PI 128216 locus on chromosome 11 provided resistance that was dependent on gene dosage and genetic background. These results define a single locus, Rx-4, from PI 128216, which provides resistance to bacterial spot race T3, has additive gene action, and is located on chromosome 11.

  2. Radicicol, a heat shock protein 90 inhibitor, inhibits differentiation and adipogenesis in 3T3-L1 preadipocytes

    Energy Technology Data Exchange (ETDEWEB)

    He, Yonghan [Department of Nutrition and Food Hygiene, Public Health College, Harbin Medical University, 157 Baojian Road, Harbin 150081 (China); Aquatic and Crop Resource Development, Life Sciences Branch, National Research Council Canada, Charlottetown, PE, Canada C1A 4P3 (Canada); State Key Laboratory of Genetic Resources and Evolution, Kunming Institute of Zoology, Chinese Academy of Sciences, Kunming 650223 (China); Li, Ying [Department of Nutrition and Food Hygiene, Public Health College, Harbin Medical University, 157 Baojian Road, Harbin 150081 (China); Zhang, Shuocheng [Aquatic and Crop Resource Development, Life Sciences Branch, National Research Council Canada, Charlottetown, PE, Canada C1A 4P3 (Canada); Perry, Ben [Aquatic and Crop Resource Development, Life Sciences Branch, National Research Council Canada, Charlottetown, PE, Canada C1A 4P3 (Canada); Department of Biomedical Sciences, University of Prince Edward Island, 550 University Avenue, Charlottetown, PE, Canada C1A 4P3 (Canada); Zhao, Tiantian [Aquatic and Crop Resource Development, Life Sciences Branch, National Research Council Canada, Charlottetown, PE, Canada C1A 4P3 (Canada); Department of Psychology, University of Toronto, 1265 Military Trail, Toronto, ON, Canada M1C 1A4 (Canada); Wang, Yanwen, E-mail: yanwen.wang@nrc.ca [Aquatic and Crop Resource Development, Life Sciences Branch, National Research Council Canada, Charlottetown, PE, Canada C1A 4P3 (Canada); Department of Biomedical Sciences, University of Prince Edward Island, 550 University Avenue, Charlottetown, PE, Canada C1A 4P3 (Canada); Sun, Changhao, E-mail: sun2002changhao@yahoo.com [Department of Nutrition and Food Hygiene, Public Health College, Harbin Medical University, 157 Baojian Road, Harbin 150081 (China)

    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.

  3. Overexpression of the mitochondrial T3 receptor p43 induces a shift in skeletal muscle fiber types.

    Directory of Open Access Journals (Sweden)

    François Casas

    Full Text Available In previous studies, we have characterized a new hormonal pathway involving a mitochondrial T3 receptor (p43 acting as a mitochondrial transcription factor and consequently stimulating mitochondrial activity and mitochondrial biogenesis. We have established the involvement of this T3 pathway in the regulation of in vitro myoblast differentiation. We have generated mice overexpressing p43 under control of the human alpha-skeletal actin promoter. In agreement with the previous characterization of this promoter, northern-blot and western-blot experiments confirmed that after birth p43 was specifically overexpressed in skeletal muscle. As expected from in vitro studies, in 2-month old mice, p43 overexpression increased mitochondrial genes expression and mitochondrial biogenesis as attested by the increase of mitochondrial mass and mt-DNA copy number. In addition, transgenic mice had a body temperature 0.8 degrees C higher than control ones and displayed lower plasma triiodothyronine levels. Skeletal muscles of transgenic mice were redder than wild-type animals suggesting an increased oxidative metabolism. In line with this observation, in gastrocnemius, we recorded a strong increase in cytochrome oxidase activity and in mitochondrial respiration. Moreover, we observed that p43 drives the formation of oxidative fibers: in soleus muscle, where MyHC IIa fibers were partly replaced by type I fibers; in gastrocnemius muscle, we found an increase in MyHC IIa and IIx expression associated with a reduction in the number of glycolytic fibers type IIb. In addition, we found that PGC-1alpha and PPARdelta, two major regulators of muscle phenotype were up regulated in p43 transgenic mice suggesting that these proteins could be downstream targets of mitochondrial activity. These data indicate that the direct mitochondrial T3 pathway is deeply involved in the acquisition of contractile and metabolic features of muscle fibers in particular by regulating PGC-1alpha

  4. T3 Regulates a Human Macrophage-Derived TSH-β Splice Variant: Implications for Human Bone Biology.

    Science.gov (United States)

    Baliram, R; Latif, R; Morshed, S A; Zaidi, M; Davies, T F

    2016-09-01

    TSH and thyroid hormones (T3 and T4) are intimately involved in bone biology. We have previously reported the presence of a murine TSH-β splice variant (TSH-βv) expressed specifically in bone marrow-derived macrophages and that exerted an osteoprotective effect by inducing osteoblastogenesis. To extend this observation and its relevance to human bone biology, we set out to identify and characterize a TSH-β variant in human macrophages. Real-time PCR analyses using human TSH-β-specific primers identified a 364-bp product in macrophages, bone marrow, and peripheral blood mononuclear cells that was sequence verified and was homologous to a human TSH-βv previously reported. We then examined TSH-βv regulation using the THP-1 human monocyte cell line matured into macrophages. After 4 days, 46.1% of the THP-1 cells expressed the macrophage markers CD-14 and macrophage colony-stimulating factor and exhibited typical morphological characteristics of macrophages. Real-time PCR analyses of these cells treated in a dose-dependent manner with T3 showed a 14-fold induction of human TSH-βv mRNA and variant protein. Furthermore, these human TSH-βv-positive cells, induced by T3 exposure, had categorized into both M1 and M2 macrophage phenotypes as evidenced by the expression of macrophage colony-stimulating factor for M1 and CCL-22 for M2. These data indicate that in hyperthyroidism, bone marrow resident macrophages have the potential to exert enhanced osteoprotective effects by oversecreting human TSH-βv, which may exert its local osteoprotective role via osteoblast and osteoclast TSH receptors.

  5. Widdrol-induced lipolysis is mediated by PKC and MEK/ERK in 3T3-L1 adipocytes.

    Science.gov (United States)

    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.

  6. Key steps in type III secretion system (T3SS) towards translocon assembly with potential sensor at plant plasma membrane.

    Science.gov (United States)

    Ji, Hongtao; Dong, Hansong

    2015-09-01

    Many plant- and animal-pathogenic Gram-negative bacteria employ the type III secretion system (T3SS) to translocate effector proteins from bacterial cells into the cytosol of eukaryotic host cells. The effector translocation occurs through an integral component of T3SS, the channel-like translocon, assembled by hydrophilic and hydrophobic proteinaceous translocators in a two-step process. In the first, hydrophilic translocators localize to the tip of a proteinaceous needle in animal pathogens, or a proteinaceous pilus in plant pathogens, and associate with hydrophobic translocators, which insert into host plasma membranes in the second step. However, the pilus needs to penetrate plant cell walls in advance. All hydrophilic translocators so far identified in plant pathogens are characteristic of harpins: T3SS accessory proteins containing a unitary hydrophilic domain or an additional enzymatic domain. Two-domain harpins carrying a pectate lyase domain potentially target plant cell walls and facilitate the penetration of the pectin-rich middle lamella by the bacterial pilus. One-domain harpins target plant plasma membranes and may play a crucial role in translocon assembly, which may also involve contrapuntal associations of hydrophobic translocators. In all cases, sensory components in the target plasma membrane are indispensable for the membrane recognition of translocators and the functionality of the translocon. The conjectural sensors point to membrane lipids and proteins, and a phosphatidic acid and an aquaporin are able to interact with selected harpin-type translocators. Interactions between translocators and their sensors at the target plasma membrane are assumed to be critical for translocon assembly.

  7. Expression of the leukemia-associated CBF{beta}/SMMHC chimeric gene causes transformation of 3T3 cells

    Energy Technology Data Exchange (ETDEWEB)

    Hajra, A.; Liu, P.; Collins, E.S. [National Center for Human Genome Research, Bethesda, MD (United States)] [and others

    1994-09-01

    A pericentric inversion of chromosome 16 (inv(16)(p13;q22)) is consistently seen in acute myeloid leukemia of the M4Eo subtype. This inversion fuses almost the entire coding region of the gene encoding of the {beta} subunit of the heterodimeric transcription factor CBF/PEBP2 to the region of the MYH11 gene encoding the rod domain for the smooth muscle myosin heavy chain (SMMHC). To investigate the biological properties of the CBF{beta}/SMMHC fusion protein, we have generated 3T3 cell lines that stably express the CBF{beta}/SMMHC chimeric cDNA or the normal, nonchimeric CBF{beta} and SMMHC cDNAs. 3T3 cells expressing CBF{beta}/SMMHC acquire a transformed phenotype, as indicated by altered cell morphology, formation of foci, and growth in soft agar. Cells constitutively overexpressing the normal CBF{beta} cDNA or the rod region of SMMHC remain nontransformed. Western blot analysis using antibodies to CBF{beta} and the SMMHC rod demonstrates that stably transfected cells express the appropriate chimeric or normal protein. Electrophoretic mobility shift assays reveal that cells transformed by the chimeric cDNA do not have a CBF-DNA complex of the expected mobility, but instead contain a large complex with CBF DNA-binding activity that fails to migrate out of the gel wells. In order to define the regions of CBF{beta}/SMMHC necessary for 3T3 transformation, we have stably transfected cells with mutant CBF{beta}/SMMHC cDNAs containing various deletions of the coding region. Analysis of these cell lines indicates that the transformation property of CBF{beta}/SMMHC requires regions of CBF{beta} known to be necessary for association with the DNA-binding CBF{alpha} subunit, and also requires an intact SMMHC carboxyl terminus, which is necessary for formation of the coiled coil domain of the myosin rod.

  8. Enhancement of ajoene-induced apoptosis by conjugated linoleic acid in 3T3-L1 adipocytes.

    Science.gov (United States)

    Yang, Jeong-Yeh; Della-Fera, Mary Anne; Hausman, Dorothy B; Baile, Clifton A

    2007-06-01

    Ajoene has been shown to induce apoptosis in 3T3-L1 adipocytes. In this report the effects on apoptosis of combinations of ajoene and trans-10, cis-12 conjugated linoleic acid (t10,c12CLA) in 3T3-L1 adipocytes were investigated. Although t10,c12CLA alone had no effect, ajoene plus t10,c12CLA reduced cell viability more than ajoene alone at 24 h (59.1 vs. 85.9% of control, respectively; p<0.05). Compared to treatment with t10,c12CLA, ajoene increased apoptosis 218% after 24 h (p<0.01), whereas ajoene plus t10,c12CLA increased apoptosis 122% over that caused by ajoene alone (p<0.01). Immunoblotting analysis also indicated that ajoene plus t10,c12CLA caused a greater increase in phosphorylation of c-Jun N-terminal kinase (JNK) and Bax expression and a greater release of mitochondrial proteins (cytochrome c, AIF) than additive responses to each compound alone. Ajoene plus t10,c12CLA also increased ROS production more than that resulting from ajoene treatment alone (264 vs 204% after 40 min, respectively; p<0.01). Furthermore, the antioxidant NAC prevented ROS generation and apoptosis by ajoene plus t10,c12CLA. Interestingly, the combination of ajoene and t10,c12CLA increased NF-kappaB activation and decreased the level of phosphorylated Akt more than each compound alone. Altogether, our observations indicate that t10,c12CLA potentiates the effect of ajoene on apoptosis in 3T3-L1 adipocytes.

  9. Effect of hexavalent chromium on proliferation and differentiation to adipocytes of 3T3-L1 fibroblasts.

    Science.gov (United States)

    Martini, Claudia N; Brandani, Javier N; Gabrielli, Matías; Vila, María del C

    2014-06-01

    Heavy metals contamination has become an important risk factor for public health and the environment. Chromium is a frequent industrial contaminant and is also used in orthopaedic joint replacements made from cobalt-chromium-alloy. Since hexavalent chromium (Cr(VI)) was reported as genotoxic and carcinogenic in different mammals, to further evaluate its cytotoxicity, we investigated the effect of this heavy metal in the proliferation and differentiation to adipocytes of 3T3-L1 fibroblasts. These cells, after the addition of a mixture containing insulin, dexamethasone and methylisobutylxanthine, first proliferate, a process known as mitotic clonal expansion (MCE), and then differentiate to adipocytes. In this differentiation process a key transcription factor is induced: peroxisome proliferator-activated receptor gamma (PPAR gamma). We found that treatment of 3T3-L1 fibroblasts with potassium chromate inhibited proliferation in exponentially growing cells and MCE as well as differentiation. A decrease in PPAR gamma content, evaluated by western blot and immunofluorescence, was found in cells differentiated in the presence of chromium. On the other hand, after inhibition of differentiation with chromium, when the metal was removed, differentiation was recovered, which indicates that this may be a reversible effect. We also found an increase in the number of micronucleated cells after treatment with Cr(VI) which is associated with genotoxic effects. According to our results, Cr(VI) is able to inhibit proliferation and differentiation to adipocytes of 3T3-L1 fibroblasts and to increase micronucleated cells, which are all indicative of alterations in cellular physiology and therefore, contributes to further elucidate the cytotoxic effects of this heavy metal. Copyright © 2014 Elsevier Ltd. All rights reserved.

  10. Comparative analysis of different feeder layers with 3T3 fibroblasts for culturing rabbits limbal stem cells

    Directory of Open Access Journals (Sweden)

    Hui-Xian Wang

    2017-07-01

    Full Text Available AIM: To explore the possibility of human umbilical cord mesenchymal stem cells (hUCMSCs, human umbilical vein endothelial cells (hUVECs, human dental pulp stem cells (hDPSCs and human periodontal ligament stem cells (hPDLSCs serving as feeder cells in co-culture systems for the cultivation of limbal stem cells. METHODS: Different feeder layers were cultured in Dulbecco’s modified Eagle’s medium (DMEM/F12 and were treated with mitomycin C. Rabbits limbal stem cells (LSCs were co-cultured on hUCMSCs, hUVECs, hDPSCs, hPDLSCs and NIH-3T3, and then comparative analysis were made between each group to see their respective colony-forming efficiency (CFE assay and immunofluorescence (IPO13,CK3/12. RESULTS: The efficiency of the four type cells in supporting the LSCs morphology and its cellular differentiation was similar to that of NIH-3T3 fibroblasts as demonstrated by the immunostaining properties analysis, with each group exhibiting a similar strong expression pattern of IPO13, but lacking CK3 and CK12 expression in terms of immunostaining. But hUCMSCs, hDPSCs and hPDLSCs feeder layers were superior in promoting colony formation potential of cells when compared to hUVECs and feeder-cell-free culture. CONCLUSION: hUCMSCs, hDPSCs and hPDLSCs can be a suitable alternative to conventional mouse NIH-3T3 feeder cells, so that risk of zoonotic infection can be diminished.

  11. Long-term outcome of hand-assisted laparoscopic nephroureterectomy for pathologic T3 upper urinary tract urothelial carcinoma.

    Science.gov (United States)

    Chung, Shiu-Dong; Chen, Shyh-Chyan; Wang, Shuo-Meng; Chueh, Shih-Chieh; Lai, Ming-Kuen; Huang, Chao-Yuan; Pu, Yeong-Shiau; Huang, Kuo-How; Yu, Hong-Jeng

    2009-01-01

    To determine the feasibility and long-term outcomes of hand-assisted laparoscopic nephroureterectomy (HALNU) compared with open nephroureterectomy (ONU) in the management of pT(3)N0 upper urinary tract urothelial carcinoma (UUT-UC). Between January 1994 and December 2005, 21 patients who underwent HALNU for stage pT(3)N0 UTT-UC were matched and compared with 31 patients who underwent ONU. The oncologic out-comes, including bladder recurrence, recurrence-free survival, cancer-specific survival, and overall survival, were statistically analyzed. The median follow-up period in the HALNU group was 72 months (range 33-111 months) and 115 months in the ONU group (range 24-161 months). Patient age, sex, body mass index, tumor size, specimen weight, and American Society of Anesthesiologists classification showed no significant difference between the two groups. The HALNU group had statistically less blood loss than the ONU group (113 mL versus 487 mL; P = 0.02). The average hospital stay and doses of narcotic analgesics were significantly less in the HALNU group than the ONU group. The complication and bladder recurrence rates were similar between the two groups. The 5-year recurrence-free survival, cancer-specific survival, and overall survival were also comparable in both groups. HALNU is a safe and efficacious procedure with comparable long-term oncologic outcomes in comparison with ONU in treating patients with locally advanced pT(3)N0UUT-UC.

  12. Analysis and Solutions of Common Problems in the Operation of Ufida T3 Software%用友T3软件操作常见问题及对策

    Institute of Scientific and Technical Information of China (English)

    程竞

    2015-01-01

    用友T3软件是会计信息化课程的常用教学软件之一。经过长期教学实践发现学生在操作中经常会出现账套号用错、演示版到期、凭证遗漏等问题,针对这些问题进行分析并探讨出通过数据库、账套输出文件及软件操作技巧等方式来有效解决的对策,供授课教师和软件学习者参考。%Ufida T3 software is one of the commonly-used softwares in accounting information course. Based on a long-term teaching experiment, the author finds out that some problems have appeared frequently, such as mistakes in accounting number, expiration of demo account number, omission of credentials, etc. This paper analyzes these problems and discusses about the effective ways to solve them through database, accounting mode file and improving software operating skills. These solutions can be the references for both teachers and learners.

  13. Divergent in vivo activity of non‐serotonergic and serotonergic VGluT3–neurones in the median raphe region

    Science.gov (United States)

    Domonkos, Andor; Nikitidou Ledri, Litsa; Laszlovszky, Tamás; Cserép, Csaba; Borhegyi, Zsolt; Papp, Edit; Nyiri, Gábor; Freund, Tamás F.

    2016-01-01

    Key points The median raphe is a key subcortical modulatory centre involved in several brain functions, such as regulation of the sleep–wake cycle, emotions and memory storage.A large proportion of median raphe neurones are glutamatergic and implement a radically different mode of communication compared to serotonergic cells, although their in vivo activity is unknown.We provide the first description of the in vivo, brain state‐dependent firing properties of median raphe glutamatergic neurones identified by immunopositivity for the vesicular glutamate transporter type 3 (VGluT3) and serotonin (5‐HT). Glutamatergic populations (VGluT3+/5‐HT– and VGluT3+/5‐HT+) were compared with the purely serotonergic (VGluT3–/5‐HT+ and VGluT3–/5‐HT–) neurones.VGluT3+/5‐HT+ neurones fired similar to VGluT3–/5‐HT+ cells, whereas they significantly diverged from the VGluT3+/5‐HT– population. Activity of the latter subgroup resembled the spiking of VGluT3–/5‐HT– cells, except for their diverging response to sensory stimulation.The VGluT3+ population of the median raphe may broadcast rapidly varying signals on top of a state‐dependent, tonic modulation. Abstract Subcortical modulation is crucial for information processing in the cerebral cortex. Besides the canonical neuromodulators, glutamate has recently been identified as a key cotransmitter of numerous monoaminergic projections. In the median raphe, a pure glutamatergic neurone population projecting to limbic areas was also discovered with a possibly novel, yet undetermined function. In the present study, we report the first functional description of the vesicular glutamate transporter type 3 (VGluT3)‐expressing median raphe neurones. Because there is no appropriate genetic marker for the separation of serotonergic (5‐HT+) and non‐serotonergic (5‐HT–) VGluT3+ neurones, we utilized immunohistochemistry after recording and juxtacellular labelling in anaesthetized rats. VGluT3+/5

  14. Downregulation of the taurine transporter TauT during hypo-osmotic stress in NIH3T3 mouse fibroblasts

    DEFF Research Database (Denmark)

    Hansen, Daniel Bloch; Friis, Martin Barfred; Hoffmann, Else Kay

    2012-01-01

    The present work was initiated to investigate regulation of the taurine transporter TauT by reactive oxygen species (ROS) and the tonicity-responsive enhancer binding protein (TonEBP) in NIH3T3 mouse fibroblasts during acute and long-term (4 h) exposure to low-sodium/hypo-osmotic stress. Taurine...... by ROS under hypo-osmotic, low-sodium conditions, whereas the TauT mRNA level is unaffected. Acute exposure to ROS reduces taurine uptake as a result of modulated TauT transport kinetics. Thus, swelling-induced ROS production could account for the reduced taurine uptake under low-sodium...

  15. HCC国航枢纽控制中心服务T3航站楼

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    3月26日,国航将进行“大搬家”,正式转入首都机场T3航站楼运营。届时,全新的HCC国航枢纽控制中心也将发挥效力,使得值机柜台的开放数目、自助值机以及航班办理手续时间等方面有较大改善和变化。

  16. Property of anodic coatings obtained in an organic, environmental friendly electrolyte on aluminum alloy 2024-T3

    Science.gov (United States)

    Zhang, R. F.; Shi, H. W.; Liu, Z. L.; Zhang, S. F.; Zhang, Y. Q.; Guo, S. B.

    2014-01-01

    Anodic coatings were obtained by micro arc oxidation on aluminum alloy 2024-T3 in a solution containing only 10 g/L sodium phytate. The morphology, composition, structure and corrosion resistance of anodic coatings were systematically investigated. The results show that the working voltage continually increases during 3 min and bright sparks appear after 25 s. Anodic coatings are evenly formed on the substrate and about 2 μm thick. XPS and XRD analyses reveal that the obtained coatings are mainly composed of α-Al2O3 and γ-Al2O3. Compared with the substrate, the corrosion resistance of the anodized aluminum alloys is improved.

  17. Trip report on IAEA Training Workshop on Implementation of Integrated Management Systems for Research Reactors (T3-TR-45496).

    Energy Technology Data Exchange (ETDEWEB)

    Pratt, Richard J.

    2013-11-01

    From 17-21 June 2013, Sandia National Laboratories, Technical Area-V (SNL TA-V) represented the United States Department of Energy/National Nuclear Security Administration (DOE/NNSA) at the International Atomic Energy Agency (IAEA) Training Workshop (T3-TR-45486). This report gives a breakdown of the IAEA regulatory structure for those unfamiliar, and the lessons learned and observations that apply to SNL TA-V that were obtained from the workshop. The Safety Report Series, IAEA workshop final report, and SNL TA-V presentation are included as attachments.

  18. Laparoscopic radical nephrectomy versus open radical nephrectomy in T1-T3 renal tumors: An outcome analysis

    Directory of Open Access Journals (Sweden)

    Arvind P Ganpule

    2008-01-01

    Full Text Available Aims: To compare laparoscopic radical nephrectomy (LRN with open radical nephrectomy (ORN in T1-T3 renal lesions. Materials and Methods: The records of 65 patients who underwent LRN between January 2002 and December 2006 were entered prospectively in a database. The patients were compared with 56 patients who had undergone ORN between January 2000 and December 2005. The two groups were comparable in terms of age, body mass index (BMI and tumor size. LRN was compared with ORN in terms of operative room time, blood loss, complications , analgesic requirement, hospital stay and start of oral intake. The oncologic efficacy was evaluated in stages T1 and T2 in terms of cancer-free and overall survival. Results: The laparoscopy group had a significantly shorter hospital stay (5.72, range 3-23 days vs. 9.18, range 4-23 days, p value: < 0.0001, analgesia requirement (175.65, range 50-550 mg vs. 236, range 0-1100 mg of tramadol, p value: < 0.03, hemoglobin decline (1.55, range 0.1 to 4.4 mg/dl vs. 2.25, range 0.2 - 7 mg/dL, p value: < 0.001 and hematocrit drop (4.83, range 0.3 - 12.9 vs. 7.06 range 2 -18, p value: < 0.0001. The majority of specimens showed renal cell carcinoma. In the laparoscopy group, 29 tumors were T1 stage, 18 were T2, while eight were T3. In the open surgery group, 25 tumors were T1, 19 were T2 and 12 were T3. The cancer-free survival rate at 24 months for ORN and LRN in T1 lesions was 91.7% and 93.15% respectively and the patient survival rate was 100% in both groups. The cancer-free survival rate at 24 months for ORN and LRN in T2 lesions was 88.9% and 94.1%, respectively and the patient survival was 100% and 94%, respectively. After LRN, there was one instance of port site metastasis, local recurrence and distant metastasis. All recurrences were distant after ORN. Conclusion: Laparoscopic radical nephrectomy has advantages in terms of shorter hospitalization and a lower analgesia requirement. It is feasible and produces effective

  19. Heat Shock Protein Augmentation of Root Hot Water Extract on Adipogenic Differentiation in Murine 3T3-L1 Preadipocytes

    Directory of Open Access Journals (Sweden)

    Wenchie Marie L. Lumbera

    2016-03-01

    Full Text Available There is a high association of heat shock on the alteration of energy and lipid metabolism. The alterations associated with thermal stress are composed of gene expression changes and adaptation through biochemical responses. Previous study showed that Angelica gigas Nakai (AGN root extract promoted adipogenic differentiation in murine 3T3-L1 preadipocytes under the normal temperature condition. However, its effect in heat shocked 3T3-L1 cells has not been established. In this study, we investigated the effect of AGN root hot water extract in the adipogenic differentiation of murine 3T3-L1 preadipocytes following heat shock and its possible mechanism of action. Thermal stress procedure was executed within the same stage of preadipocyte confluence (G0 through incubation at 42°C for one hour and then allowed to recover at normal incubation temperature of 37°C for another hour before AGN treatment for both cell viability assay and Oil Red O. Cell viability assay showed that AGN was able to dose dependently (0 to 400 μg/mL increase cell proliferation under normal incubation temperature and also was able to prevent cytotoxicity due to heat shock accompanied by cell proliferation. Confluent preadipocytes were subjected into heat shock procedure, recovery and then AGN treatment prior to stimulation with the differentiation solution. Heat shocked preadipocytes exhibited reduced differentiation as supported by decreased amount of lipid accumulation in Oil Red O staining and triglyceride measurement. However, those heat shocked preadipocytes that then were given AGN extract showed a dose dependent increase in lipid accumulation as shown by both evaluation procedures. In line with these results, real-time polymerase chain reaction (RT-PCR and Western blot analysis showed that AGN increased adipogenic differentiation by upregulating heat shock protection related genes and proteins together with the adipogenic markers. These findings imply the potential of

  20. The Pharmacological Effects of Morroniside and Loganin Isolated from Liuweidihuang Wan, on MC3T3-E1 Cells

    OpenAIRE

    Xijun Wang; Hui Sun; Ping Wang; Kun Yang; Wei Wang; Manyu Li

    2010-01-01

    Liuweidihuang Wan (LW), initially a well-known formula for curing “wu chi wu ruan”, is commonly used nowadays for clinical treatment of Postmenopausal Osteoporosis (PO), but the identity of the effective substance(s) remains unclear. The present study was designed to evaluate the effects of morroniside and loganin isolated from LW on the proliferation, differentiation and apoptosis of MC3T3-E1 cells, as well as the possible mechanism of action. Morroniside and loganin had no effects on the pr...

  1. Inhibitory effects of LPA1 on cell motile activities stimulated by hydrogen peroxide and 2,3-dimethoxy-1,4-naphthoquinone in fibroblast 3T3 cells.

    Science.gov (United States)

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

    2013-11-08

    Reactive oxygen species (ROS) are known to mediate a variety of biological responses, including cell motility. Recently, we indicated that lysophosphatidic acid (LPA) receptor-3 (LPA3) increased cell motile activity stimulated by hydrogen peroxide. In the present study, we assessed the role of LPA1 in the cell motile activity mediated by ROS in mouse fibroblast 3T3 cells. 3T3 cells were treated with hydrogen peroxide and 2,3-dimethoxy-1,4-naphthoquinone (DMNQ) at concentrations of 0.1 and 1 μM for 48 h. In cell motility assays with Cell Culture Inserts, the cell motile activities of 3T3 cells treated with hydrogen peroxide and DMNQ were significantly higher than those of untreated cells. 3T3 cells treated with hydrogen peroxide and DMNQ showed elevated expression levels of the Lpar3 gene, but not the Lpar1 and Lpar2 genes. To investigate the effects of LPA1 on the cell motile activity induced by hydrogen peroxide and DMNQ, Lpar1-overexpressing (3T3-a1) cells were generated from 3T3 cells and treated with hydrogen peroxide and DMNQ. The cell motile activities stimulated by hydrogen peroxide and DMNQ were markedly suppressed in 3T3-a1 cells. These results suggest that LPA signaling via LPA1 inhibits the cell motile activities stimulated by hydrogen peroxide and DMNQ in 3T3 cells.

  2. The anti-apoptotic MAP kinase pathway is inhibited in NIH3T3 fibroblasts with increased expression of phosphatidylinositol transfer protein β

    NARCIS (Netherlands)

    Schenning, M.; van Tiel, C.M.; Wirtz, K.W.A.; Snoek, G.T.

    2007-01-01

    Mouse NIH3T3 fibroblast cells overexpressing phosphatidylinositol transfer protein ß (PI-TPß, SPIß cells) demonstrate a low rate of proliferation and a high sensitivity towards UV-induced apoptosis when compared with wtNIH3T3 cells. In contrast, SPIßS262A cells overexpressing a mutant PI-TPß that la

  3. Dehydrodiconiferyl alcohol isolated from Cucurbita moschata shows anti-adipogenic and anti-lipogenic effects in 3T3-L1 cells and primary mouse embryonic fibroblasts.

    Science.gov (United States)

    Lee, Junghun; Kim, Donghyun; Choi, Jonghyun; Choi, Hyounjeong; Ryu, Jae-Ha; Jeong, Jinhyun; Park, Eun-Jin; Kim, Seon-Hee; Kim, Sunyoung

    2012-03-16

    A water-soluble extract from the stems of Cucurbita moschata, code named PG105, was previously found to contain strong anti-obesity activities in a high fat diet-induced obesity mouse model. One of its biological characteristics is that it inhibits 3T3-L1 adipocyte differentiation. To isolate the biologically active compound(s), conventional solvent fractionation was performed, and the various fractions were tested for anti-adipogenic activity using Oil Red O staining method. A single spot on thin layer chromatography of the chloroform fraction showed a potent anti-adipogenic activity. When purified, the structure of its major component was resolved as dehydrodiconiferyl alcohol (DHCA), a lignan, by NMR and mass spectrometry analysis. In 3T3-L1 cells, synthesized DHCA significantly reduced the expression of several adipocyte marker genes, including peroxisome proliferator-activated receptor γ (Pparg), CCAAT/enhancer-binding protein α (Cebpa), fatty acid-binding protein 4 (Fabp4), sterol response element-binding protein-1c (Srebp1c), and stearoyl-coenzyme A desaturase-1 (Scd), and decreased lipid accumulation without affecting cell viability. DHCA also suppressed the mitotic clonal expansion of preadipocytes (an early event of adipogenesis), probably by suppressing the DNA binding activity of C/EBPβ, and lowered the production level of cyclinA and cyclin-dependent kinase 2 (Cdk2), coinciding with the decrease in DNA synthesis and cell division. In addition, DHCA directly inhibited the expression of SREBP-1c and SCD-1. Similar observations were made, using primary mouse embryonic fibroblasts. Taken together, our data indicate that DHCA may contain dual activities, affecting both adipogenesis and lipogenesis.

  4. 中华鳖对T3菌苗抗原的免疫应答%IMMUNE RESPONSE OF SOFT-SHELLED TURTLE (TRIONYX SINENSIS) TO BACTERIN T3

    Institute of Scientific and Technical Information of China (English)

    杨先乐; 周剑光; 艾晓辉; 柯福恩

    2001-01-01

    The immune response in soft-shelled turtle (Trionyx sinensis) immunized wit h bacterin T3, was determined with indirect agglutination titer (IAT) and perc ent relative protection (PRP). In the vaccinated turtle, the strong immune respo nse to bacterin T3 was produced. Six groups of 1-year-old turtles and 19 gro ups of 2-years-old turtles were respectively vaccinated by bacterin T3. At the 30th day, the PRP and IAT of the 1-year-old turtles were to 618±112(6)% and 4033±1654(6), yet the corresponding values of 2-years-old turtles were 826±152(19)% and 1?9781±7164(19) respectively. Acc or ding to the test, the PRP and IAT of 2-years-old turtles were higher than thos e of 1-year-old turtles. The study demonstrated that the effective phase of im mune response to bacterin T3 was 3 days. At the 20th day, the IAT value is up to 1?768 0±4476(4). Which maintained for about 10 day s and decreased to the initial level on the 60thday. The results proved th at the immunologic memory of the turtles was strong. The secondary enhanced immu n ization began at the 30th day of the primary immunization. The IAT increa sed quickly and was 5?7600±2?7900(4) at the 16th day of the seco n dary immunization, which reached the second peak value that was nine times as hi gh as the primary peak value. The IAT decreased to 4?2667 ±1?4780(3) at the 30th day, 2?9865±1?9553(3) at the 60th da y, and 1? 2800±0(2) at the 70th day. The result still has no significant differen ce compared to the peak value of the secondary immunization (02<P<03), yet having significantly higher than that of immunization (001<P<002) . From these facts, we can concluded that the decreasing speed of antibody of th e secondary immunization was clearly slower than increasing speed. The result sug g ests that the character of immune response of Soft

  5. Effects and Molecular Mechanism of GST-Irisin on Lipolysis and Autocrine Function in 3T3-L1 Adipocytes.

    Directory of Open Access Journals (Sweden)

    Shanshan Gao

    Full Text Available Irisin, which was recently identified as a myokine and an adipokine, transforms white adipose tissue to brown adipose tissue and has increasingly caught the attention of the medical and scientific community. However, the signaling pathway of irisin and the molecular mechanisms responsible for the lipolysis effect remain unclear. In this study, we established an efficient system for the expression and purification of GST-irisin in Escherichia coli. The biological activity of GST-irisin was verified using the cell counting kit-8 assay and by detecting the mRNA expression of uncoupling protein 1. Our data showed that GST-irisin regulates mRNA levels of lipolysis-related genes such as adipose triglyceride lipase and hormone-sensitive lipase and proteins such as the fatty acid-binding protein 4, leading to increased secretion of glycerol and decreased lipid accumulation in 3T3-L1 adipocytes. In addition, exogenous GST-irisin can increase its autocrine function in vitro by regulating the expression of fibronectin type III domain-containing protein 5. GST-irisin could regulate glucose uptake in 3T3-L1 adipocytes. Hence, we believe that recombinant GST-irisin could promote lipolysis and its secretion in vitro and can potentially prevent obesity and related metabolic diseases.

  6. Radioimmunoassay of a human serum growth factor for Balb/c-3T3 cells: derivation from platelets.

    Science.gov (United States)

    Antoniades, H N; Scher, C D

    1977-05-01

    A radioimmunoassay has been developed for the detection and quantification of a human serum polypeptide that has growth-promoting activity for confluent Balb/c-3T3 cells. Antiserum to this growth factor does not recognize antigens in mouse, guinea pig, or bovine serum but does detect some crossreacting antigen in the serum of the New World monkey Cebus albifrons and more in the serum of the Old World rhesus monkeys Macaca mulatta and M. fascicularis, demonstrating that the antigenic determinants of the growth factor have a degree of species specificity. Serum derived from whole human blood contains approximately 770 pg of the growth factor per mg of protein; serum derived from platelet-poor blood contains about 112 pg of the growth factor per mg of protein. As much as 1 microng of the growth factor per mg of protein has been recovered from human platelets by heating them at 100 degrees for 2 min. Approximately 1-2 ng of the growth factor, in either whole serum or platelets, stimulates 5 to 10 X 10(3) confluent Balb/c-3T3 cells to replicate. The heat treatment of platelets allows the purification and quantitative recovery of the growth factor from blood.

  7. Resveratrol metabolites modify adipokine expression and secretion in 3T3-L1 pre-adipocytes and mature adipocytes.

    Directory of Open Access Journals (Sweden)

    Itziar Eseberri

    Full Text Available OBJECTIVE: Due to the low bioavailability of resveratrol, determining whether its metabolites exert any beneficial effect is an interesting issue. METHODS: 3T3-L1 maturing pre-adipocytes were treated during differentiation with 25 µM of resveratrol or with its metabolites and 3T3-L1 mature adipocytes were treated for 24 hours with 10 µM resveratrol or its metabolites. The gene expression of adiponectin, leptin, visfatin and apelin was assessed by Real Time RT-PCR and their concentration in the incubation medium was quantified by ELISA. RESULTS: Resveratrol reduced mRNA levels of leptin and increased those of adiponectin. It induced the same changes in leptin secretion. Trans-resveratrol-3-O-glucuronide and trans-resveratrol-4'-O-glucuronide increased apelin and visfatin mRNA levels. Trans-resveratrol-3-O-sulfate reduced leptin mRNA levels and increased those of apelin and visfatin. CONCLUSIONS: The present study shows for the first time that resveratrol metabolites have a regulatory effect on adipokine expression and secretion. Since resveratrol has been reported to reduce body-fat accumulation and to improve insulin sensitivity, and considering that these effects are mediated in part by changes in the analyzed adipokines, it may be proposed that resveratrol metabolites play a part in these beneficial effects of resveratrol.

  8. Ethanolic extract of Actaea racemosa (black cohosh) potentiates bone nodule formation in MC3T3-E1 preosteoblast cells.

    Science.gov (United States)

    Chan, B Y; Lau, K S; Jiang, B; Kennelly, E J; Kronenberg, F; Kung, A W C

    2008-09-01

    Aceaea racemosa (formerly Cimicifuga racemosa, black cohosh, AR) extracts have been widely used as an alternative to hormonal replacement therapy for menopausal symptoms. Recent evidences suggest AR extracts are also effective in protecting against postmenopausal bone loss. To determine whether AR has any direct anabolic effect on osteoblasts, we investigated the ethanolic extract of AR on bone nodule formation in mouse MC3T3-E1 preosteoblast cells. AR did not stimulate osteoblast proliferation. Rather, at high doses of 1000 ng/mL for 48 h, AR suppressed (7.2+/-0.9% vs. control) osteoblast proliferation. At 500 ng/mL, a significant increase in bone nodule formation was seen with Von Kossa staining. Using quantitative PCR analysis, AR was shown to enhance the gene expression of runx2 and osteocalcin. Co-treatment with ICI 182,780, the selective estrogen receptor antagonist, abolished the stimulatory effect of AR on runx2 and osteocalcin gene induction, as well as on bone nodule formation in MC3T3-E1 cells. This is a first report of the direct effect of AR on enhancement of bone nodule formation in osteoblasts, and this action was mediated via an estrogen receptor-dependent mechanism. The results provide a scientific rationale at the molecular level for the claim that AR can offer effective prevention of postmenopausal bone loss.

  9. Three-dimensional reconstruction of the Shigella T3SS transmembrane regions reveals 12-fold symmetry and novel features throughout

    Energy Technology Data Exchange (ETDEWEB)

    Hodgkinson, J.L.; Wall, J.; Horsley, A.; Stabat, S.; Simon, M.; Johnson, S.; da Fonseca, P. C. A.; Morris, E. P.; Lea, S. M.; Blocker, A. J.

    2009-05-18

    Type III secretion systems (T3SSs) mediate bacterial protein translocation into eukaryotic cells, a process essential for virulence of many Gram-negative pathogens. They are composed of a cytoplasmic secretion machinery and a base that bridges both bacterial membranes, into which a hollow, external needle is embedded. When isolated, the latter two parts are termed the 'needle complex'. An incomplete understanding of the structure of the needle complex has hampered studies of T3SS function. To estimate the stoichiometry of its components, we measured the mass of its subdomains by scanning transmission electron microscopy (STEM). We determined subunit symmetries by analysis of top and side views within negatively stained samples in low-dose transmission electron microscopy (TEM). Application of 12-fold symmetry allowed generation of a 21-25-{angstrom} resolution, three-dimensional reconstruction of the needle complex base, revealing many new features and permitting tentative docking of the crystal structure of EscJ, an inner membrane component.

  10. Benzyl butyl phthalate promotes adipogenesis in 3T3-L1 preadipocytes: A High Content Cellomics and metabolomic analysis.

    Science.gov (United States)

    Yin, Lei; Yu, Kevin Shengyang; Lu, Kun; Yu, Xiaozhong

    2016-04-01

    Benzyl butyl phthalate (BBP) has been known to induce developmental and reproductive toxicity. However, its association with dysregulation of adipogenesis has been poorly investigated. The present study aimed to examine the effect of BBP on the adipogenesis, and to elucidate the underlying mechanisms using the 3T3-L1 cells. The capacity of BBP to promote adipogenesis was evaluated by multiple staining approaches combined with a High Content Cellomics analysis. The dynamic changes of adipogenic regulatory genes and proteins were examined, and the metabolite profile was identified using GC/MC based metabolomic analysis. The High Content analysis showed BBP in contrast with Bisphenol A (BPA), a known environmental obesogen, increased lipid droplet accumulation in a similar dose-dependent manner. However, the size of the lipid droplets in BBP-treated cells was significantly larger than those in cells treated with BPA. BBP significantly induced mRNA expression of transcriptional factors C/EBPα and PPARγ, their downstream genes, and numerous adipogenic proteins in a dose and time-dependent manner. Furthermore, GC/MC metabolomic analysis revealed that BBP exposure perturbed the metabolic profiles that are associated with glyceroneogenesis and fatty acid synthesis. Altogether, our current study clearly demonstrates that BBP promoted the differentiation of 3T3-L1 through the activation of the adipogenic pathway and metabolic disturbance.

  11. Synthesis of polypyrrole nanowires with positive effect on MC3T3-E1 cell functions through electrical stimulation.

    Science.gov (United States)

    He, Yuan; Wang, Shihui; Mu, Jing; Dai, Lingfeng; Zhang, Zhong; Sun, Yanan; Shi, Wei; Ge, Dongtao

    2017-02-01

    Conducting polymer polypyrrole (PPy) possesses good biocompatibility and conductivity and has been used as functional coatings in bone tissue regeneration. In this study, a cholic acid doped PPy nanowires (PPy NWs) coating was electrochemically polymerized on the surface of titanium (Ti). The porous intertwined PPy NWs coating exhibited excellent electrical conductivity and electrochemical activity, better hydrophilicity and higher surface energy. In vitro cell experiments demonstrated that the PPy NWs coating together with a 10μA substrate-mediate electrical stimulation (ES) was capable to positive regulate the functions of MC3T3-E1 such as cell adhesion, proliferation and differentiation. Further long-term functions of cell tests including alkaline phosphatase (ALP) activity, bone-carboxyglutamic acid-containing protein (BGP) and calcium deposition were all thoroughly increased. These confirmed that the combination of PPy NWs and ES could accelerate MC3T3-E1 cells mature and osteogenesis. Hence, the PPy NWs coating was an electro bioactive coating and may have potential applications in the treatment of bone damage repairing and regeneration with ES. Copyright © 2016. Published by Elsevier B.V.

  12. Altered gene expression profiles of NIH3T3 cells regulated by human lung cancer associated gene CT120

    Institute of Scientific and Technical Information of China (English)

    Xiang Huo HE; Jin Jun LI; Yi Hu XIE; Yun Tian TANG; Gen Fu YAO; Wen Xin QIN; Da Fang WAN; Jian Ren GU

    2004-01-01

    CT120, a novel membrane-associated gene implicated in lung carcinogenesis, was previously identified from chromosome 17p13.3 locus, a hot mutation spot involved in human malignancies. In the present study, we further determined that CT120 ectopic expression could promote cell proliferation activity of NIH3T3 cells using MTS assay, and monitored the downstream effects of CT120 in NIH3T3 cells with Atlas mouse cDNA expression arrays. Among 588known genes, 133 genes were found to be upregulated or downregulated by CT120. Two major signaling pathways involved in cell proliferation, cell survival and anti-apoptosis were overexpressed and activated in response to CT120:One is the Raf/MEK/Erk signal cascades and the other is the PI3K/Akt signal cascades, suggesting that CT120 might contribute, at least in part, to the constitutively activation of Erk and Akt in human lung caner cells. In addition, some tumor metastasis associated genes cathepsin B, cathepsin D, cathepsin L, MMP-2/TIMP-2 were also upregulated by CT120, upon which CT120 might be involved in tumor invasiveness and metastasis. In addition, CT120 might play an important role in tumor progression through modulating the expression of some candidate "Lung Tumor Progression"genes including B-Raf, Rab-2, BAX, BAG-1, YB-1, and Cdc42.

  13. Investigation on the corrosion behaviour and microstructure of 2024-T3 Al alloy treated via plasma electrolytic oxidation

    Energy Technology Data Exchange (ETDEWEB)

    Fadaee, Hossein; Javidi, Mehdi, E-mail: mjavidi@yahoo.com

    2014-08-01

    Highlights: • Plasma electrolytic oxidation used to improve corrosion resistance of 2024-T3 alloy. • A conventional DC source with low applied voltage was considered for PEO. • Use of glycerin was effective in enhancement of corrosion resistance. • The surface morphology indicated coatings have a dense structure without cracking. • Degradation of coatings with immersion time was investigated. - Abstract: Plasma electrolytic oxidation treatment was performed on 2024-T3 aluminium alloy in a simple and inexpensive electrolyte. It was found that treatment time of 30 min resulted in the most noble corrosion potential and lowest corrosion current density. The surface and cross-sectional morphology indicated that the coating has a dense structure without any cracking. Furthermore, the result of X-ray diffraction analysis indicated that the coating was formed mainly from α-Al{sub 2}O{sub 3}, γ-Al{sub 2}O{sub 3}, Al{sub 3.21}Si{sub 0.47} and small amounts of amorphous phases. The presence of glycerine in the electrolyte composition resulted in not only stabilising the solution but also improving the corrosion resistance.

  14. Suppression of lipin-1 expression increases monocyte chemoattractant protein-1 expression in 3T3-L1 adipocytes

    Energy Technology Data Exchange (ETDEWEB)

    Takahashi, Nobuhiko, E-mail: ntkhs@hoku-iryo-u.ac.jp [Department of Internal Medicine, School of Dentistry, Health Sciences University of Hokkaido, 1757 Kanazawa, Ishikari-Toubetsu, Hokkaido 061-0023 (Japan); Division of Gastroenterology and Hematology/Oncology, Department of Medicine, Asahikawa Medical University, 2-1-1-1 Midorigaoka-Higashi, Asahikawa, Hokkaido 078-8510 (Japan); Yoshizaki, Takayuki [Innovation Center, Kagoshima University, 1-21-40 Korimoto, Kagoshima 890-0065 (Japan); Hiranaka, Natsumi; Suzuki, Takeshi [Department of Internal Medicine, School of Dentistry, Health Sciences University of Hokkaido, 1757 Kanazawa, Ishikari-Toubetsu, Hokkaido 061-0023 (Japan); Yui, Tomoo; Akanuma, Masayasu; Oka, Kazuya [Department of Fixed Prosthodontics and Oral Implantology, School of Dentistry, Health Sciences University of Hokkaido, 1757 Kanazawa, Ishikari-Toubetsu, Hokkaido 061-0023 (Japan); Kanazawa, Kaoru [Department of Dental Anesthesiology, School of Dentistry, Health Sciences University of Hokkaido, 1757 Kanazawa, Ishikari-Toubetsu, Hokkaido 061-0023 (Japan); Yoshida, Mika; Naito, Sumiyoshi [Department of Clinical Laboratory, Health Sciences University of Hokkaido, 1757 Kanazawa, Ishikari-Toubetsu, Hokkaido 061-0023 (Japan); Fujiya, Mikihiro; Kohgo, Yutaka [Division of Gastroenterology and Hematology/Oncology, Department of Medicine, Asahikawa Medical University, 2-1-1-1 Midorigaoka-Higashi, Asahikawa, Hokkaido 078-8510 (Japan); Ieko, Masahiro [Department of Internal Medicine, School of Dentistry, Health Sciences University of Hokkaido, 1757 Kanazawa, Ishikari-Toubetsu, Hokkaido 061-0023 (Japan)

    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.

  15. Astragaloside IV attenuates lipolysis and improves insulin resistance induced by TNFalpha in 3T3-L1 adipocytes.

    Science.gov (United States)

    Jiang, Boren; Yang, Ying; Jin, Hua; Shang, Wenbin; Zhou, Libin; Qian, Lei; Chen, Mingdao

    2008-11-01

    Increased circulating free fatty acid (FFA) concentrations have been demonstrated to potentially link obesity, insulin resistance and cardiovascular diseases. Astragaloside IV (AS-IV) is a saponin which is widely used in traditional Chinese medicine to treat type 2 diabetes and cardiovascular diseases. The purpose of the present study was to examine the effects of AS-IV on the lipolysis and insulin resistance induced by tumor necrosis factor-alpha (TNFalpha) in cultured 3T3-L1 adipocytes. TNFalpha promotes lipolysis in mammal adipocytes via the mitogen activated protein kinase (MAPK) family resulting in reduced expression/function of perilipin. Application of AS-IV inhibited TNFalpha-induced accelerated lipolysis in a dose-dependent manner, which was compatible with suppressed phosphorylation of ERK1/2 and reversed the downregulation of perilipin. Moreover, TNFalpha induced downregulation of key enzymes in lipogenesis, including LPL, FAS and GPAT, were also attenuated by AS-IV. Further studies showed that AS-IV improved TNFalpha-induced insulin resistance in 3T3-L1 adipocytes. This study provides the first direct evidence of the antilipolytic action of AS-IV in adipocytes, which may allow this agent to decrease the circulating FFA levels, thus increase insulin sensitivity and treat cardiovascular diseases.

  16. Hmgb1 promotes wound healing of 3T3 mouse fibroblasts via RAGE-dependent ERK1/2 activation.

    Science.gov (United States)

    Ranzato, Elia; Patrone, Mauro; Pedrazzi, Marco; Burlando, Bruno

    2010-05-01

    HMGb1 is a nuclear protein playing a role in DNA architecture and transcription. This protein has also been shown to function as a cytokine and to stimulate keratinocyte scratch wound healing. Due to the importance of finding new wound healing molecules, we have studied the effects of HMGb1 on fibroblasts, another major skin cell type, using the NIH 3T3 line. HMGb1 expression in these cells was assessed by Western blot, while its nuclear localization was pointed out by confocal immunofluorescence. HMGb1-induced cell proliferation with a maximum at a concentration of 10 nM, and such a dose also stimulated cell migration and scratch wound healing. Western blot analysis showed that HMGb1 activates ERK1/2, while the use of an anti-RAGE receptor-blocking antibody and of the selective MEK1/2 inhibitor PD98059 blocked ERK1/2 activation and wound healing responses to HMGb1. Taken together data show that HMGb1 promotes 3T3 fibroblast wound healing by inducing cell proliferation and migration, and that this occurs through the activation of the RAGE/MEK/ERK pathway. In conclusion, HMGb1 seems a good candidate for the development of medical treatments to be used on chronic or severe wounds.

  17. A Nanodot Array Modulates Cell Adhesion and Induces an Apoptosis-Like Abnormality in NIH-3T3 Cells

    Directory of Open Access Journals (Sweden)

    Hung Yao-Ching

    2009-01-01

    Full Text Available Abstract Micro-structures that mimic the extracellular substratum promote cell growth and differentiation, while the cellular reaction to a nanostructure is poorly defined. To evaluate the cellular response to a nanoscaled surface, NIH 3T3 cells were grown on nanodot arrays with dot diameters ranging from 10 to 200 nm. The nanodot arrays were fabricated by AAO processing on TaN-coated wafers. A thin layer of platinum, 5 nm in thickness, was sputtered onto the structure to improve biocompatibility. The cells grew normally on the 10-nm array and on flat surfaces. However, 50-nm, 100-nm, and 200-nm nanodot arrays induced apoptosis-like events. Abnormality was triggered after as few as 24 h of incubation on a 200-nm dot array. For cells grown on the 50-nm array, the abnormality started after 72 h of incubation. The number of filopodia extended from the cell bodies was lower for the abnormal cells. Immunostaining using antibodies against vinculin and actin filament was performed. Both the number of focal adhesions and the amount of cytoskeleton were decreased in cells grown on the 100-nm and 200-nm arrays. Pre-coatings of fibronectin (FN or type I collagen promoted cellular anchorage and prevented the nanotopography-induced programed cell death. In summary, nanotopography, in the form of nanodot arrays, induced an apoptosis-like abnormality for cultured NIH 3T3 cells. The occurrence of the abnormality was mediated by the formation of focal adhesions.

  18. 3T3-L1 preadipocytes exhibit heightened monocyte-chemoattractant protein-1 response to acute fatty acid exposure.

    Science.gov (United States)

    Dordevic, Aimee L; Konstantopoulos, Nicky; Cameron-Smith, David

    2014-01-01

    Preadipocytes contribute to the inflammatory responses within adipose tissue. Whilst fatty acids are known to elicit an inflammatory response within adipose tissue, the relative contribution of preadipocytes and mature adipocytes to this is yet to be determined. We aimed to examine the actions of common dietary fatty acids on the acute inflammatory and adipokine response in 3T3-L1 preadipocytes and differentiated mature adipocytes. Gene expression levels of key adipokines in 3T3-L1 preadipocytes and adipocytes were determined following incubation with palmitic acid, myristic acid or oleic acid and positive inflammatory control, lipopolysaccharide for 2 and 4 h. Inflammatory kinase signalling was assessed by analysis of nuclear factor-κB, p38-mitogen-activated protein kinase and c-jun amino-terminal kinase phosphorylation. Under basal conditions, intracellular monocyte chemoattractant protein-1 and interleukin-6 gene expression levels were increased in preadipocytes, whereas mature adipocytes expressed increased gene expression levels of leptin and adiponectin. Fatty acid exposure at 2 and 4 h increased both monocyte chemoattractant protein-1 and interleukin-6 gene expression levels in preadipocytes to greater levels than in mature adipocytes. There was an accompanying increase of inhibitor of κB-α degradation and nuclear factor-κB (p65) (Ser536) phosphorylation with fatty acid exposure in the preadipocytes only. The current study points to preadipocytes rather than the adipocytes as the contributors to both immune cell recruitment and inflammatory adipokine secretion with acute increases in fatty acids.

  19. Curcumin attenuates lipolysis stimulated by tumor necrosis factor-α or isoproterenol in 3T3-L1 adipocytes.

    Science.gov (United States)

    Xie, Xiao-yun; Kong, Po-Ren; Wu, Jin-feng; Li, Ying; Li, Yan-xiang

    2012-12-15

    Curcumin, an active component derived from dietary spice turmeric (Curcuma longa), has been demonstrated antihyperglycemic, antiinflammatory and hypocholesterolemic activities in obesity and diabetes. These effects are associated with decreased level of circulating free fatty acids (FFA), however the mechanism has not yet been elucidated. The flux of FFA and glycerol from adipose tissue to the blood stream primarily depends on the lipolysis of triacylglycerols in the adipocytes. Adipocyte lipolysis is physiologically stimulated by catecholamine hormones. Tumor necrosis factor-α (TNFα) stimulates chronic lipolysis in obesity and type 2 diabetes. In this study, we examined the role of curcumin in inhibiting lipolytic action upon various stimulations in 3T3-L1 adipocytes. Glycerol release from TNFα or isoproterenol-stimulated 3T3-L1 adipocytes in the absence or presence of curcumin was determined using a colorimetric assay (GPO-Trinder). Western blotting was used to investigate the TNFα-induced phosphorylation of MAPK and perilipin expression. Fatcake and cytosolic fractions were prepared to examine the isoproterenol-stimulated hormone-sensitive lipase translocation. Treatment with curcumin attenuated TNFα-mediated lipolysis by suppressing phosphorylation of extracellular signal-related kinase 1/2 (ERK1/2) and reversing the downregulation of perilipin protein in TNFα-stimulated adipocytes (plipolysis response to TNFα and catecholamines. The antilipolytic effect could be a cellular basis for curcumin decreasing plasma FFA levels and improving insulin sensitivity. Copyright © 2012 Elsevier GmbH. All rights reserved.

  20. Catechin and quercetin attenuate adipose inflammation in fructose-fed rats and in 3T3-L1 adipocytes

    Science.gov (United States)

    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