Cross sections for 12C+12C→12C(0+2)+12C(g.s.) using breathing mode doorways

International Nuclear Information System (INIS)

Ahmed, M.U.; Beres, W.P.

1982-01-01

A previously derived projection operator method is applied to the calculation of the cross section for 12 C+ 12 C→ 12 C(0 + 2 )+ 12 C(g.s.) with a breathing mode model being used to describe the 0 + 2 (7.68 MeV) state of 12 C. The relationship to processes leading to alpha particle channels is discussed. The cross section for 12 C+ 12 C→ 12 C(3 - )+ 12 C(g.s.) is also calculated and possible correlations with inelastic scattering to the 0 + 2 and 2 + states of 12 C are discussed. The results for both 0 + 2 and 3 - inelastic scattering are in reasonable agreement with experiment

Bioenergetic Profile Experiment using C2C12 Myoblast Cells

Science.gov (United States)

Nicholls, David G.; Darley-Usmar, Victor M.; Wu, Min; Jensen, Per Bo; Rogers, George W.; Ferrick, David A.

2010-01-01

The ability to measure cellular metabolism and understand mitochondrial dysfunction, has enabled scientists worldwide to advance their research in understanding the role of mitochondrial function in obesity, diabetes, aging, cancer, cardiovascular function and safety toxicity. Cellular metabolism is the process of substrate uptake, such as oxygen, glucose, fatty acids, and glutamine, and subsequent energy conversion through a series of enzymatically controlled oxidation and reduction reactions. These intracellular biochemical reactions result in the production of ATP, the release of heat and chemical byproducts, such as lactate and CO2 into the extracellular environment. Valuable insight into the physiological state of cells, and the alteration of the state of those cells, can be gained through measuring the rate of oxygen consumed by the cells, an indicator of mitochondrial respiration - the Oxygen Consumption Rate - or OCR. Cells also generate ATP through glycolysis, i.e.: the conversion of glucose to lactate, independent of oxygen. In cultured wells, lactate is the primary source of protons. Measuring the lactic acid produced indirectly via protons released into the extracellular medium surrounding the cells, which causes acidification of the medium provides the Extra-Cellular Acidification Rate - or ECAR. In this experiment, C2C12 myoblast cells are seeded at a given density in Seahorse cell culture plates. The basal oxygen consumption (OCR) and extracellular acidification (ECAR) rates are measured to establish baseline rates. The cells are then metabolically perturbed by three additions of different compounds (in succession) that shift the bioenergetic profile of the cell. This assay is derived from a classic experiment to assess mitochondria and serves as a framework with which to build more complex experiments aimed at understanding both physiologic and pathophysiologic function of mitochondria and to predict the ability of cells to respond to stress and

Mechanical stimulation of C2C12 cells increases m-calpain expression and activity, focal adhesion plaque degradation and cell fusion

DEFF Research Database (Denmark)

Grossi, Alberto; Karlsson, Anders Hans; Lawson, Moira A.

2005-01-01

Abstract Mechanical Stimulation of C2C12 Cells Increases m-calpain Expression and Activity, Focal Adhesion Plaque Degradation and Cell Fusion A. Grossi, A. H. Karlsson, M. A. Lawson; Department of Dairy and Food Science, Royal Veterinary and Agricultural University, Frederiksberg C, Denmark...... Myogenesis is a complex sequence of events, including the irreversible transition from the proliferation-competent myoblast stage into fused, multinucleated myotubes. During embryonic development, myogenic differentiation is regulated by positive and negative signals from surrounding tissues. Stimulation due...... to the activity of ubiquitous proteolytic enzymes known as calpains has been reported. Whether there is a link between stretch- or load induced signaling and calpain expression and activation is not known. Using a magnetic bead stimulation assay and C2C12 mouse myoblasts cell population, we have demonstrated...

DRAGON, a GPI-anchored membrane protein, inhibits BMP signaling in C2C12 myoblasts.

Science.gov (United States)

Kanomata, Kazuhiro; Kokabu, Shoichiro; Nojima, Junya; Fukuda, Toru; Katagiri, Takenobu

2009-06-01

Bone morphogenetic proteins (BMPs) induce osteoblastic differentiation of myoblasts via binding to cell surface receptors. Repulsive guidance molecules (RGMs) have been identified as BMP co-receptors. We report here that DRAGON/RGMb, a member of the RGM family, suppressed BMP signaling in C2C12 myoblasts via a novel mechanism. All RGMs were expressed in C2C12 cells that were differentiated into myocytes and osteoblastic cells, but RGMc was not detected in immature cells. In C2C12 cells, only DRAGON suppressed ALP and Id1 promoter activities induced by BMP-4 or by constitutively activated BMP type I receptors. This inhibition by DRAGON was dependent on the secretory form of the von Willbrand factor type D domain. DRAGON even suppressed BMP signaling induced by constitutively activated Smad1. Over-expression of neogenin did not alter the inhibitory capacity of DRAGON. Taken together, these findings indicate that DRAGON may be an inhibitor of BMP signaling in C2C12 myoblasts. We also suggest that a novel molecule(s) expressed on the cell membrane may mediate the signal transduction of DRAGON in order to suppress BMP signaling in C2C12 myoblasts.

Actin-associated protein palladin is required for migration behavior and differentiation potential of C2C12 myoblast cells

Energy Technology Data Exchange (ETDEWEB)

Nguyen, Ngoc Uyen Nhi; Liang, Vincent Roderick; Wang, Hao-Ven, E-mail: hvwang@mail.ncku.edu.tw

2014-09-26

Highlights: • Palladin is involved in myogenesis in vitro. • Palladin knockdown by siRNA increases myoblast proliferation, viability and differentiation. • Palladin knockdown decreases C2C12 myoblast migration ability. - Abstract: The actin-associated protein palladin has been shown to be involved in differentiation processes in non-muscle tissues. However, but its function in skeletal muscle has rarely been studied. Palladin plays important roles in the regulation of diverse actin-related signaling in a number of cell types. Since intact actin-cytoskeletal remodeling is necessary for myogenesis, in the present study, we pursue to investigate the role of actin-associated palladin in skeletal muscle differentiation. Palladin in C2C12 myoblasts is knocked-down using specific small interfering RNA (siRNA). The results show that down-regulation of palladin decreased migratory activity of mouse skeletal muscle C2C12 myoblasts. Furthermore, the depletion of palladin enhances C2C12 vitality and proliferation. Of note, the loss of palladin promotes C2C12 to express the myosin heavy chain, suggesting that palladin has a role in the modulation of C2C12 differentiation. It is thus proposed that palladin is required for normal C2C12 myogenesis in vitro.

Actin-associated protein palladin is required for migration behavior and differentiation potential of C2C12 myoblast cells

International Nuclear Information System (INIS)

Nguyen, Ngoc Uyen Nhi; Liang, Vincent Roderick; Wang, Hao-Ven

2014-01-01

Highlights: • Palladin is involved in myogenesis in vitro. • Palladin knockdown by siRNA increases myoblast proliferation, viability and differentiation. • Palladin knockdown decreases C2C12 myoblast migration ability. - Abstract: The actin-associated protein palladin has been shown to be involved in differentiation processes in non-muscle tissues. However, but its function in skeletal muscle has rarely been studied. Palladin plays important roles in the regulation of diverse actin-related signaling in a number of cell types. Since intact actin-cytoskeletal remodeling is necessary for myogenesis, in the present study, we pursue to investigate the role of actin-associated palladin in skeletal muscle differentiation. Palladin in C2C12 myoblasts is knocked-down using specific small interfering RNA (siRNA). The results show that down-regulation of palladin decreased migratory activity of mouse skeletal muscle C2C12 myoblasts. Furthermore, the depletion of palladin enhances C2C12 vitality and proliferation. Of note, the loss of palladin promotes C2C12 to express the myosin heavy chain, suggesting that palladin has a role in the modulation of C2C12 differentiation. It is thus proposed that palladin is required for normal C2C12 myogenesis in vitro

Zein nanoparticle as a novel BMP6 derived peptide carrier for enhanced osteogenic differentiation of C2C12 cells.

Science.gov (United States)

Hadavi, Mahvash; Hasannia, Sadegh; Faghihi, Shahab; Mashayekhi, Farhad; Homazadeh, Homayoun; Mostofi, Seyed Behrooz

2018-01-26

Zein nanoparticles as a carrier system for BMP6-derived peptide were prepared by liquid-liquid phase separation procedure and characterized with SEM, DLS, FTIR and thermogravimetric methods. After peptide encapsulation, nanoparticle size increased from 236.3 ± 92.2 nm to 379.4 ± 116.8 nm. The encapsulation efficiency of peptide was 72.6% and the release of peptide from Zein nanoparticles was partly sustained in trypsin containing phosphate buffered saline (pH 7.4) for up to 14 days. Peptide-loaded nanoparticles showed similar cell viability compared with blank ones. ALP activity of C2C12 cells treated with peptide-loaded nanoparticles (500 µg/mL) was evaluated 7, 14, 21 and 28 days after culture. In peptide-loaded nanoparticles, ALP activity was significantly higher (p < .05) compared with other groups at day 14. Alizarin Red S staining showed, C2C12 cells behind peptide-loaded nanoparticles had significantly (p < .05) higher calcium deposition at day 21. The results of RT-qPCR show that the BMP-6 peptide activated expression of RUNX2 as a transcription factor. In turn, RUNX2 regulates SPP1 and BGLAP gene expression, as osteogenic marker genes. The results confirm that the peptide-loaded Zein nanoparticles, as osteoinductive material, may be used to repair small area of bone defects, with low load bearing.

Conditions Inducing Excessive O-GlcNAcylation Inhibit BMP2-Induced Osteogenic Differentiation of C2C12 Cells.

Science.gov (United States)

Gu, Hanna; Song, Mina; Boonanantanasarn, Kanitsak; Baek, Kyunghwa; Woo, Kyung Mi; Ryoo, Hyun-Mo; Baek, Jeong-Hwa

2018-01-09

Hyperglycemic conditions in diabetic patients can affect various cellular functions, including the modulation of osteogenic differentiation. However, the molecular mechanisms by which hyperglycemia affects osteogenic differentiation are yet to be clarified. This study aimed to investigate whether the aberrant increase in protein O -linked-β- N -acetylglucosamine glycosylation ( O -GlcNAcylation) contributes to the suppression of osteogenic differentiation due to hyperglycemia. To induce osteogenic differentiation, C2C12 cells were cultured in the presence of recombinant human bone morphogenetic protein 2 (BMP2). Excessive protein O -GlcNAcylation was induced by treating C2C12 cells with high glucose, glucosamine, or N -acetylglucosamine concentrations or by O -GlcNAc transferase (OGT) overexpression. The effect of O -GlcNAcylation on osteoblast differentiation was then confirmed by examining the expression levels of osteogenic marker gene mRNAs, activity of alkaline phosphatase, and transcriptional activity of Runx2, a critical transcription factor for osteoblast differentiation and bone formation. Cell treatment with high glucose, glucosamine or N -acetylglucosamine increased O -GlcNAcylation of Runx2 and the total levels of O -GlcNAcylated proteins, which led to a decrease in the transcriptional activity of Runx2, expression levels of osteogenic marker genes (Runx2, osterix, alkaline phosphatase, and type I collagen), and activity of alkaline phosphatase. These inhibitory effects were rescued by lowering protein O -GlcNAcylation levels by adding STO45849, an OGT inhibitor, or by overexpressing β- N -acetylglucosaminidase. Our findings suggest that excessive protein O -GlcNAcylation contributes to high glucose-suppressed osteogenic differentiation.

BAMBI Promotes C2C12 Myogenic Differentiation by Enhancing Wnt/β-Catenin Signaling

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Qiangling Zhang

2015-08-01

Full Text Available Bone morphogenic protein and activin membrane-bound inhibitor (BAMBI is regarded as an essential regulator of cell proliferation and differentiation that represses transforming growth factor-β and enhances Wnt/β-catenin signaling in various cell types. However, its role in skeletal muscle remains largely unknown. In the current study, we found that the expression level of BAMBI peaked in the early differentiation phase of the C2C12 rodent myoblast cell line. Knockdown of BAMBI via siRNA inhibited C2C12 differentiation, indicated by repressed MyoD, MyoG, and MyHC expression as well as reductions in the differentiation and fusion indices. BAMBI knockdown reduced the activity of Wnt/β-catenin signaling, as characterized by the decreased nuclear translocation of β-catenin and the lowered transcription of Axin2, which is a well-documented target gene of the Wnt/β-catenin signaling pathway. Furthermore, treatment with LiCl, an activator of Wnt/β-catenin signaling, rescued the reduction in C2C12 differentiation caused by BAMBI siRNA. Taken together, our data suggest that BAMBI is required for normal C2C12 differentiation, and that its role in myogenesis is mediated by the Wnt/β-catenin pathway.

Effect of Curcumin on Phosphorylation of AMPK and ACC in C2C12 Skeletal Muscle Cells

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F Ghanbarzadeh

2013-10-01

Full Text Available Introduction: AMP activated protein kinase (AMPK as key regulators of cell metabolism, plays a major role in the activation of catabolic pathways, such as glucose transport and fatty acid oxidation. Thus, activation of this pathway can be used in the treatment of diabetes and metabolic syndrome. Many studied proposed the effectiveness of the polyphenols present in rhizomes of turmeric (curcumin on diabetes and its related complications. Therefore, this study investigated the effects of curcumin as an activator of AMPK pathway in C2C12 muscle cells. Methods: This study was done on C2C12 skeletal muscle cell line. The cells were classified into two distinct groups: first group was treated with 40µM curcumin and the second one with 0.1% DMSO as a negative control. The phosphorylated (AMPK and phosphorylated acetyl COA carboxylase (ACC were evaluated and compared by Western blotting technique. Results: intracellular phosphorylated AMPK protein content in Curcumin-treated group was 132.6% and ACC protein phosphorylated was 366.47%. Conclusion: This study showed that the levels of phosphorylated AMPK and ACC protein in cells treated with curcumin are higher than the negative control. Thus curcumin can be regarded as an activator of AMPK activity in these cells and can assist as a potential target for making anti diabetic medecine that has a synergistic activity with insulin.

Graphene-Based Patterning and Differentiation of C2C12 Myoblasts

DEFF Research Database (Denmark)

Bajaj, Piyush; Rivera, Jose A; Marchwiany, Daniel

2014-01-01

This study aims at generating highly aligned functional myotubes using graphene as the underlying scaffold. Graphene not only supports the growth of C2C12 muscle cells but also enhances its differentiation and leads to spontaneous patterning of myotubes....

Effect of curcumin Extract on Ttranslocation of Glut 4 in C2C12 Myotubes

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J Zavarreza

2013-06-01

Full Text Available Introduction: Curcumin is a major phenolic compound of Curcuma longa, which has long been used in traditional Indian medicine. Recently, curcumin has been reported to have antihyperglycemic activity in animal models. However, the molecular basis of this action has not been adequatedly described. In the present study the antihyperglycemic effect of curcumin was examined using C2C12 myoblast cells. Methods: The effects of curcumin were investigated in C2C12 myotubes by treating the cells with 40 µM of curcumin for 1.5 h. C2C12 myotubes were homogenized and the subcellular fractionation was prepared using ultracentrifugation; Then protein assay was performed using Bradford method and Glut4 determination was done using SDS-PAGE. Moreover, western immunoblotting techniques were exerted for semi-quantitative measurement. Data analysis was performed via gene tools software of Gel documentation and SPSS. An ANOVA test was used to compare three groups together. Results: Comparison of Glut4 levels in C2C12 myotubes showed that myotubes which were exposed to1.5 hours of 40 µM curcunin had higher Glut4 percentages in both cytosolic and membrane fractions and Glut4 percentages were significant with a confidence interval (CI of 95% ( P<0.05 . Conclusion: The study results showed that curcumin can strongly induce the increase of Glut4 translocation in differentiated C2C12 cells, indicating its possible regulatory role in the glucose metabolism of skeletal muscle cells

First intron of nestin gene regulates its expression during C2C12 myoblast ifferentiation

Institute of Scientific and Technical Information of China (English)

Hua Zhong; Zhigang Jin; Yongfeng Chen; Ting Zhang; Wei Bian; Xing Cui; Naihe Jing

2008-01-01

Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China Nestin is an intermediate filament protein expressed in neural progenitor cells and in developing skeletal muscle. Nestin has been widely used as a neural progenitor cell marker. It is well established that the specific expression of the nestin gene in neural progenitor cells is conferred by the neural-specific enhancer located in the second intron of the nestin gene. However, the transcriptional mechanism of nestin expression in developing muscle is still unclear. In this study, we identified a muscle cell-specific enhancer in the first intron of mouse nestin gene in mouse myoblast C2C12 cells.We localized the core enhancer activity to the 291-661 region of the first intron, and showed that the two E-boxes in the core enhancer region were important for enhancer activity in differentiating C2C12 cells. We also showed that MyoD protein was involved in the regulation of nestin expression in the myogenic differentiation of C2C12 cells.

Correlation between the 12C+12C, 12C+13C, and 13C+13C fusion cross sections

Science.gov (United States)

Notani, M.; Esbensen, H.; Fang, X.; Bucher, B.; Davies, P.; Jiang, C. L.; Lamm, L.; Lin, C. J.; Ma, C.; Martin, E.; Rehm, K. E.; Tan, W. P.; Thomas, S.; Tang, X. D.; Brown, E.

2012-01-01

The fusion cross section for 12C+13C has been measured down to Ec.m.=2.6 MeV, at which the cross section is of the order of 20 nb. By comparing the cross sections for the three carbon isotope systems, 12C+12C, 12C+13C, and 13C+13C, it is found that the cross sections for 12C+13C and 13C+13C provide an upper limit for the fusion cross section of 12C+12C over a wide energy range. After calibrating the effective nuclear potential for 12C+12C using the 12C+13C and 13C+13C fusion cross sections, it is found that a coupled-channels calculation with the ingoing wave boundary condition (IWBC) is capable of predicting the major peak cross sections in 12C+12C. A qualitative explanation for this upper limit is provided by the Nogami-Imanishi model and by level density differences among the compound nuclei. It is found that the strong resonance found at 2.14 MeV in 12C+12C exceeds this upper limit by a factor of more than 20. The preliminary result from the most recent measurement shows a much smaller cross section at this energy, which agrees with our predicted upper limit.

Thimerosal-induced apoptosis in mouse C2C12 myoblast cells occurs through suppression of the PI3K/Akt/survivin pathway.

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Wen-Xue Li

Full Text Available BACKGROUND: Thimerosal, a mercury-containing preservative, is one of the most widely used preservatives and found in a variety of biological products. Concerns over its possible toxicity have reemerged recently due to its use in vaccines. Thimerosal has also been reported to be markedly cytotoxic to neural tissue. However, little is known regarding thimerosal-induced toxicity in muscle tissue. Therefore, we investigated the cytotoxic effect of thimerosal and its possible mechanisms on mouse C2C12 myoblast cells. METHODOLOGY/PRINCIPAL FINDINGS: The study showed that C2C12 myoblast cells underwent inhibition of proliferation and apoptosis after exposure to thimerosal (125-500 nM for 24, 48 and 72 h. Thimerosal caused S phase arrest and induced apoptosis as assessed by flow cytometric analysis, Hoechst staining and immunoblotting. The data revealed that thimerosal could trigger the leakage of cytochrome c from mitochondria, followed by cleavage of caspase-9 and caspase-3, and that an inhibitor of caspase could suppress thimerosal-induced apoptosis. Thimerosal inhibited the phosphorylation of Akt(ser473 and survivin expression. Wortmannin, a PI3K inhibitor, inhibited Akt activity and decreased survivin expression, resulting in increased thimerosal-induced apoptosis in C2C12 cells, while the activation of PI3K/Akt pathway by mIGF-I (50 ng/ml increased the expression of survivin and attenuated apoptosis. Furthermore, the inhibition of survivin expression by siRNA enhanced thimerosal-induced cell apoptosis, while overexpression of survivin prevented thimerosal-induced apoptosis. Taken together, the data show that the PI3K/Akt/survivin pathway plays an important role in the thimerosal-induced apoptosis in C2C12 cells. CONCLUSIONS/SIGNIFICANCE: Our results suggest that in C2C12 myoblast cells, thimerosal induces S phase arrest and finally causes apoptosis via inhibition of PI3K/Akt/survivin signaling followed by activation of the mitochondrial apoptotic

Caspase activity and apoptotic signaling in proliferating C2C12 cells following cisplatin or A23187 exposure

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Darin Bloemberg

2016-06-01

Full Text Available Investigating cell death signaling using cell culture is commonly performed to examine the effects of novel pharmaceuticals or to further characterize discrete cellular signaling pathways. Here, we provide data regarding the cell death response to either cisplatin or A23187 in sub-confluent C2C12 cells, by utilizing several concentrations and incubation times for each chemical. These data include an assessment of the activation of the proteolytic enzymes caspase-3, caspase-8, caspase-9, calpain, and cathepsin B/L. Additionally, the expression of the apoptosis-regulating proteins Bax, Bcl2, and p53 are presented.

Classification of C2C12 cells at differentiation by convolutional neural network of deep learning using phase contrast images.

Science.gov (United States)

Niioka, Hirohiko; Asatani, Satoshi; Yoshimura, Aina; Ohigashi, Hironori; Tagawa, Seiichi; Miyake, Jun

2018-01-01

In the field of regenerative medicine, tremendous numbers of cells are necessary for tissue/organ regeneration. Today automatic cell-culturing system has been developed. The next step is constructing a non-invasive method to monitor the conditions of cells automatically. As an image analysis method, convolutional neural network (CNN), one of the deep learning method, is approaching human recognition level. We constructed and applied the CNN algorithm for automatic cellular differentiation recognition of myogenic C2C12 cell line. Phase-contrast images of cultured C2C12 are prepared as input dataset. In differentiation process from myoblasts to myotubes, cellular morphology changes from round shape to elongated tubular shape due to fusion of the cells. CNN abstract the features of the shape of the cells and classify the cells depending on the culturing days from when differentiation is induced. Changes in cellular shape depending on the number of days of culture (Day 0, Day 3, Day 6) are classified with 91.3% accuracy. Image analysis with CNN has a potential to realize regenerative medicine industry.

Morphometric studies with attached mouse C3H/10T 1/2 cells

International Nuclear Information System (INIS)

Geard, C.R.; Harding, T.

1981-01-01

Studies of in vitro transformation using the Syrian hamster embryo cell system and the mouse C3H/10T 1/2 cell system form an integral part of this laboratory's activities. As part of the studies with the mouse cell line we have monitored the behavior of these cells in culture in order to ascertain those variables which might influence the expression of transformation. The study of transformed cells versus normal cells could lead to insight into an earlier definition of transformation that the clonal morphological change currently in use. This present report details the changes in cellular morphology with time in culture of normal mouse C3H/10T 1/2 cells from early passages (9 to 13) and x-ray transformed cells which have been maintained in culture for three years

IGF-1 prevents simvastatin-induced myotoxicity in C2C12 myotubes.

Science.gov (United States)

Bonifacio, Annalisa; Sanvee, Gerda M; Brecht, Karin; Kratschmar, Denise V; Odermatt, Alex; Bouitbir, Jamal; Krähenbühl, Stephan

2017-05-01

Statins are generally well tolerated, but treatment with these drugs may be associated with myopathy. The mechanisms of statin-associated myopathy are not completely understood. Statins inhibit AKT phosphorylation by an unclear mechanism, whereas insulin-like growth factor (IGF-1) activates the IGF-1/AKT signaling pathway and promotes muscle growth. The aims of the study were to investigate mechanisms of impaired AKT phosphorylation by simvastatin and to assess effects of IGF-1 on simvastatin-induced myotoxicity in C2C12 myotubes. C2C12 mouse myotubes were exposed to 10 μM simvastatin and/or 10 ng/mL IGF-1 for 18 h. Simvastatin inhibited the IGF-1/AKT signaling pathway, resulting in increased breakdown of myofibrillar proteins, impaired protein synthesis and increased apoptosis. Simvastatin inhibited AKT S473 phosphorylation, indicating reduced activity of mTORC2. In addition, simvastatin impaired stimulation of AKT T308 phosphorylation by IGF-1, indicating reduced activation of the IGF-1R/PI3K pathway by IGF-1. Nevertheless, simvastatin-induced myotoxicity could be at least partially prevented by IGF-1. The protective effects of IGF-1 were mediated by activation of the IGF-1R/AKT signaling cascade. Treatment with IGF-1 also suppressed muscle atrophy markers, restored protein synthesis and inhibited apoptosis. These results were confirmed by normalization of myotube morphology and protein content of C2C12 cells exposed to simvastatin and treated with IGF-1. In conclusion, impaired activity of AKT can be explained by reduced function of mTORC2 and of the IGF-1R/PI3K pathway. IGF-1 can prevent simvastatin-associated cytotoxicity and metabolic effects on C2C12 cells. The study gives insight into mechanisms of simvastatin-associated myotoxicity and provides potential targets for therapeutic intervention.

Cartap-induced cytotoxicity in mouse C2C12 myoblast cell line and the roles of calcium ion and oxidative stress on the toxic effects.

Science.gov (United States)

Liao, Jiunn-Wang; Kang, Jaw-Jou; Jeng, Chian-Ren; Chang, Shao-Kuang; Kuo, Ming-Jang; Wang, Shun-Cheng; Liu, Michael R S; Pang, Victor Fei

2006-02-15

Our previous study has demonstrated that instead of neuromuscular blockage cartap, an organonitrogen insecticide, could cause a marked irreversible Ca2+-dependent contracture in both isolated mouse and rabbit phrenic nerve-diaphragms. We further examined the potential of direct myocytotoxicity of cartap and the possible roles of calcium ion and oxidative stress on cartap-induced muscle cell injury using the mouse myoblast cell line, C2C12. Cartap exerted a dose- and time-dependent cytotoxic effect in C2C12 cells measured by MTT colorimetric assay and trypan blue dye exclusion. The extracellular activities of both creatine kinase (CK) and lactate dehydrogenase (LDH) were elevated in the cartap-treated groups at or greater than 100 microM. The isoenzymatic profiles showed that the elevations were mainly due to CK-3, LDH-3, and LDH-4. Following the addition of 0.5-2.5mM EGTA, a Ca2+ chelator, or 30-100 microM verapamil, an L-type Ca2+ channel blocker, the cartap-induced reduction in MTT metabolic rate of C2C12 cells was significantly restored in a dose-dependent manner in both EGTA and verapamil-treated cells. Furthermore, EGTA could significantly reduce the cartap-induced elevation in the levels of total extracellular CK and LDH activities. Additionally, cartap significantly increased the level of endogenous reactive oxygen species (ROS) in C2C12 cells in a dose- and time-dependent manner. The cartap-induced ROS generation could be significantly inhibited by antioxidants, including Vitamins C and E, catalase, and superoxide dismutase, with catalase the most effective. EGTA could significantly inhibit cartap-induced ROS generation in a dose-dependent manner. The results suggested that cartap could induce ROS generation in C2C12 cells via a Ca2+-dependent mechanism resulting in subsequent cytotoxicity, at least partially, to C2C12 cells. It is speculated that both Ca2+ and Ca2+-induced ROS may also play the central role on the myogenic contracture and myofiber injury

17β-Estradiol Protects Mitochondrial Functions through Extracellular-Signal-Regulated Kinase in C2C12 Muscle Cells

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Ana C. Ronda

2013-10-01

Full Text Available Background/Aims: We have previously shown that exposure to 17β-estradiol (E2 prior to induction of apoptosis with H2O2 protects skeletal muscle cells against oxidative damage. However, the mechanism involved in the protective action of the hormone is poorly understood. In the present study, we focused on the mechanism by which ERK mediates this survival effect in connection with COXIV activity and mitochondrial membrane potential. Methods: Immunocytochemistry, Western blot, cytochrome c oxidase complex IV (COXIV activity, coimmunoprecipitation and JC-1 dye by flow cytometry were carried out using C2C12 myoblasts as experimental model. Results: E2 is able to activate ERK and then induces its translocation to mitochondria. Using the pharmacological inhibitor of ERK activation U0126 we show that E2, through ERK activation, is able to enhance COXIV activity. Moreover, the hormone increases the interaction between COXIV and ERK. Also, we found that hydrogen peroxide decreases COXIV activity and that preincubation of the cells with E2 prior to induction of apoptosis prevents this effect. In addition, we observe that the estrogen inhibits the collapse of mitochondrial membrane potential induced by H2O2, involving ERK and COXIV. Conclusion: Our data demonstrate that E2 promotes ERK activation and translocation to mitochondria preventing the decline in COXIV activity and in turn, alteration of mitochondrial membrane potential by oxidative stress, in C2C12 myoblasts.

Partial inelasticity coefficients of negative pions in p, d, α, 12C + 12C and p, 12C + 181Ta collisions at 4.2 GeV/c per nucleon

International Nuclear Information System (INIS)

Olimov, K.K.; Olimov, K.; Gulamov, K.G.; Olimov, A.K.; Lutpullaev, S.L.; Yuldashev, B.S.; Haseeb, M.Q.

2015-01-01

The partial inelasticity coefficients of the negative pions were determined in minimum bias p, d, α, 12 C + 12 C and p, 12 C + 18 Ta collisions at 4.2A GeV/c taking into account the average number of participant nucleons of a projectile nucleus. In nucleus–nucleus collisions, the average values of partial inelasticity coefficients (〈K(π-)〉) of the negative pions did not depend on the mass numbers of projectile and target nuclei. Increase of 〈K(π - )〉 in going from p + 12 C to d, α, 12 C + 12 C collisions was due to an additional source of production of fast negative pions in nucleus–nucleus collisions — a charge exchange conversion of one or more neutrons of a projectile nucleus into a proton and π - . Linking the experimental results of the present analysis at intermediate energy with those obtained at high and ultra-high energies, it was concluded that the average values of partial inelasticity coefficients of pions in nucleon–nucleus and nucleus–nucleus collisions manifest a transitive behavior. At intermediate energies, the values of 〈K(π - )〉 were smaller by a factor of two and more as compared to those at high energies, and they increased further with increasing incident energy, reaching a plateau at E 0 > 100A GeV. (author)

C1-2 arthrography

Energy Technology Data Exchange (ETDEWEB)

Chevrot, A [Service de Radiologie B, Hopital Cochin, 75 - Paris (France); Cermakova, E [Service de Radiologie B, Hopital Cochin, 75 - Paris (France); Vallee, C [Service de Radiologie B, Hopital Cochin, 75 - Paris (France); Chancelier, M D [Service de Radiologie B, Hopital Cochin, 75 - Paris (France); Chemla, N [Service de Radiologie B, Hopital Cochin, 75 - Paris (France); Rousselin, B [Service de Radiologie B, Hopital Cochin, 75 - Paris (France); Langer-Cherbit, A [Service de Radiologie B, Hopital Cochin, 75 - Paris (France)

1995-08-01

One hundred patients with the following conditions were studied: cervical pain or neuralgia without radiographic changes, osteoarthritis, rheumatoid arthritis, ankylosing spondylarthritis and diverse conditions. The technique consists of lateral puncture of the posterior aspect of the C1-2 joint with a 20-gauge needle under fluoroscopic control, arthrography using 1 ml contrast medium, and a 1-ml long-acting steroid injection subsequently. The articular cavity has an anterior and a posterior recess. Sometimes the posterior recess is large. In 18% of cases the contralateral joint also opacifies. C1-2 arthrography appears to be an efficient and safe technique for the treatment of upper cervical pain due to C1-2 articular disorders. (orig.)

C1-2 arthrography

International Nuclear Information System (INIS)

Chevrot, A.; Cermakova, E.; Vallee, C.; Chancelier, M.D.; Chemla, N.; Rousselin, B.; Langer-Cherbit, A.

1995-01-01

One hundred patients with the following conditions were studied: cervical pain or neuralgia without radiographic changes, osteoarthritis, rheumatoid arthritis, ankylosing spondylarthritis and diverse conditions. The technique consists of lateral puncture of the posterior aspect of the C1-2 joint with a 20-gauge needle under fluoroscopic control, arthrography using 1 ml contrast medium, and a 1-ml long-acting steroid injection subsequently. The articular cavity has an anterior and a posterior recess. Sometimes the posterior recess is large. In 18% of cases the contralateral joint also opacifies. C1-2 arthrography appears to be an efficient and safe technique for the treatment of upper cervical pain due to C1-2 articular disorders. (orig.)

Characterization of an acute muscle contraction model using cultured C2C12 myotubes.

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Yasuko Manabe

Full Text Available A cultured C2C12 myotube contraction system was examined for application as a model for acute contraction-induced phenotypes of skeletal muscle. C2C12 myotubes seeded into 4-well rectangular plates were placed in a contraction system equipped with a carbon electrode at each end. The myotubes were stimulated with electric pulses of 50 V at 1 Hz for 3 ms at 997-ms intervals. Approximately 80% of the myotubes were observed to contract microscopically, and the contractions lasted for at least 3 h with electrical stimulation. Calcium ion (Ca²⁺ transient evoked by the electric pulses was detected fluorescently with Fluo-8. Phosphorylation of protein kinase B/Akt (Akt, 5' AMP-activated protein kinase (AMPK, p38 mitogen-activated protein kinase (p38, and c-Jun NH2-terminal kinase (JNK1/2, which are intracellular signaling proteins typically activated in exercised/contracted skeletal muscle, was observed in the electrically stimulated C2C12 myotubes. The contractions induced by the electric pulses increased glucose uptake and depleted glycogen in the C2C12 myotubes. C2C12 myotubes that differentiated after exogenous gene transfection by a lipofection or an electroporation method retained their normal contractile ability by electrical stimulation. These findings show that our C2C12 cell contraction system reproduces the muscle phenotypes that arise invivo (exercise, in situ (hindlimb muscles in an anesthetized animal, and invitro (dissected muscle tissues in incubation buffer by acute muscle contraction, demonstrating that the system is applicable for the analysis of intracellular events evoked by acute muscle contraction.

Heavy ion irradiation induces autophagy in irradiated C2C12 myoblasts and their bystander cells

International Nuclear Information System (INIS)

Hino, Mizuki; Tajika, Yuki; Hamada, Nobuyuki

2010-01-01

Autophagy is one of the major processes involved in the degradation of intracellular materials. Here, we examined the potential impact of heavy ion irradiation on the induction of autophagy in irradiated C2C12 mouse myoblasts and their non-targeted bystander cells. In irradiated cells, ultrastructural analysis revealed the accumulation of autophagic structures at various stages of autophagy (id est (i.e.) phagophores, autophagosomes and autolysosomes) within 20 min after irradiation. Multivesicular bodies (MVBs) and autolysosomes containing MVBs (amphisomes) were also observed. Heavy ion irradiation increased the staining of microtubule-associated protein 1 light chain 3 and LysoTracker Red (LTR). Such enhanced staining was suppressed by an autophagy inhibitor 3-methyladenine. In addition to irradiated cells, bystander cells were also positive with LTR staining. Altogether, these results suggest that heavy ion irradiation induces autophagy not only in irradiated myoblasts but also in their bystander cells. (author)

Efeitos do ultra-som terapêutico contínuo sobre a proliferação e viabilidade de células musculares C2C12 Effects of continuous therapeutic ultrasound on proliferation and viability of C2C12 muscle cells

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Paola Pelegrineli Artilheiro

2010-06-01

Full Text Available O ultra-som terapêutico (US é um recurso bioestimulante utilizado para propiciar reparo muscular de melhor qualidade e menor duração, mas o potencial terapêutico do US contínuo não está totalmente estabelecido. O objetivo deste trabalho foi avaliar o efeito do US contínuo sobre a proliferação e viabilidade de células musculares precursoras (mioblastos C2C12. Mioblastos C2C12 foram cultivados em meio de cultura contendo 10% de soro fetal bovino e irradiados com US contínuo nas freqüências de 1 e 3 MHz nas intensidades de 0,2 e 0,5 W/cm2, durante 2 e 5 minutos. A viabilidade e proliferação celular foram avaliadas após 24, 48 e 72 h de incubação. Grupos não-irradiados serviram como controle. Foram realizados experimentos independentes em cada condição acima, e os dados obtidos submetidos à análise estatística. Os resultados mostram que não houve diferença estatisticamente significativa na proliferação e viabilidade celular entre os mioblastos tratados com US e as culturas controles após os diferentes períodos de incubação, em todos os parâmetros avaliados. Conclui-se que o US contínuo, nos parâmetros avaliados, não foi capaz de alterar a proliferação e viabilidade dos mioblastos.Therapeutic ultrasound (US is a biophysical stimulation resource widely used in order to promote better, faster muscle repair, but the effectiveness of continuous US in treating injuries is not fully established. The aim of the present in vitro study was to assess the effects of continuous ultrasound on viability and proliferation of skeletal muscle precursor cells (C2C12 myoblasts. C2C12 myoblasts were cultured in a medium containing 10% foetal bovine serum and irradiated with continuous ultrasound at 1 and 3 MHz frequencies, at intensities of 0.2 and 0.5 W/cm² for 2 and 5 minutes. Cell viability and proliferation were assessed after different incubation periods (24, 48 and 72 h. Non-irradiated groups served as control and data were

Wnt/β-catenin signaling changes C2C12 myoblast proliferation and differentiation by inducing Id3 expression

International Nuclear Information System (INIS)

Zhang, Long; Shi, Songting; Zhang, Juan; Zhou, Fangfang; Dijke, Peter ten

2012-01-01

Highlights: ► Expression of Id3 but not Id1 is induced by Wnt3a stimulation in C2C12 cells. ► Wnt3a induces Id3 expression via canonical Wnt/β-catenin pathway. ► Wnt3a-induced Id3 expression does not depend on BMP signaling activation. ► Induction of Id3 expression is critical determinant in Wnt3a-induced cell proliferation and differentiation. -- Abstract: Canonical Wnt signaling plays important roles in regulating cell proliferation and differentiation. In this study, we report that inhibitor of differentiation (Id)3 is a Wnt-inducible gene in mouse C2C12 myoblasts. Wnt3a induced Id3 expression in a β-catenin-dependent manner. Bone morphogenetic protein (BMP) also potently induced Id3 expression. However, Wnt-induced Id3 expression occurred independent of the BMP/Smad pathway. Functional studies showed that Id3 depletion in C2C12 cells impaired Wnt3a-induced cell proliferation and alkaline phosphatase activity, an early marker of osteoblast cells. Id3 depletion elevated myogenin induction during myogenic differentiation and partially impaired Wnt3a suppressed myogenin expression in C2C12 cells. These results suggest that Id3 is an important Wnt/β-catenin induced gene in myoblast cell fate determination.

NF-κB mediates the transcription of mouse calsarcin-1 gene, but not calsarcin-2, in C2C12 cells

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Mu Yulian

2007-03-01

Full Text Available Abstract Background The calsarcins comprise a novel family of muscle-specific calcineurin-interaction proteins that play an important role in modulating both the function and substrate specificity of calcineurin in muscle cells. The expression of calsarcin-1 (CS-1 is restricted to slow-twitch skeletal muscle fibres, whereas that of both calsarcin-2 (CS-2 and calsarcin-3 (CS-3 is enriched in fast-twitch fibres. However, the transcriptional control of this selective expression has not been previously elucidated. Results Our real-time RT-PCR analyses suggest that the expression of CS-1 and CS-2 is increased during the myogenic differentiation of mouse C2C12 cells. Promoter deletion analysis further suggests that an NF-κB binding site within the CS-1 promoter is responsible for the up-regulation of CS-1 transcription, but no similar mechanism was evident for CS-2. These findings are further supported by the results of EMSA analysis, as well as by overexpression and inhibition experiments in which NF-κB function was blocked by treatment with its inhibitor, PDTC. In addition, the overexpression of NFATc4 induces both the CS-1 and CS-2 promoters, whereas MEF2C only activates CS-1. Conclusion Our present data suggest that NF-κB is required for the transcription of mouse CS-1 but not CS-2, and that the regulation of the calsarcins is mediated also by the NFAT and MEF2 transcription factors. These results provide new insights into the molecular mechanisms governing transcription in specific muscle fibre cells. The calsarcins may also serve as a valuable mechanistic tool to better understand the regulation of calcineurin signalling during muscle differentiation.

PDGF-induced migration of synthetic vascular smooth muscle cells through c-Src-activated L-type Ca2+ channels with full-length CaV1.2 C-terminus.

Science.gov (United States)

Guo, Xiaoguang; Kashihara, Toshihide; Nakada, Tsutomu; Aoyama, Toshifumi; Yamada, Mitsuhiko

2018-06-01

In atherosclerosis, vascular smooth muscle cells (VSMC) migrate from the media toward the intima of the arteries in response to cytokines, such as platelet-derived growth factor (PDGF). However, molecular mechanism underlying the PDGF-induced migration of VSMCs remains unclear. The migration of rat aorta-derived synthetic VSMCs, A7r5, in response to PDGF was potently inhibited by a Ca V 1.2 channel inhibitor, nifedipine, and a Src family tyrosine kinase (SFK)/Abl inhibitor, bosutinib, in a less-than-additive manner. PDGF significantly increased Ca V 1.2 channel currents without altering Ca V 1.2 protein expression levels in A7r5 cells. This reaction was inhibited by C-terminal Src kinase, a selective inhibitor of SFKs. In contractile VSMCs, the C-terminus of Ca V 1.2 is proteolytically cleaved into proximal and distal C-termini (PCT and DCT, respectively). Clipped DCT is noncovalently reassociated with PCT to autoinhibit the channel activity. Conversely, in synthetic A7r5 cells, full-length Ca V 1.2 (Ca V 1.2FL) is expressed much more abundantly than truncated Ca V 1.2. In a heterologous expression system, c-Src activated Ca V 1.2 channels composed of Ca V 1.2FL but not truncated Ca V 1.2 (Ca V 1.2Δ1763) or Ca V 1.2Δ1763 plus clipped DCT. Further, c-Src enhanced the coupling efficiency between the voltage-sensing domain and activation gate of Ca V 1.2FL channels by phosphorylating Tyr1709 and Tyr1758 in PCT. Compared with Ca V 1.2Δ1763, c-Src could more efficiently bind to and phosphorylate Ca V 1.2FL irrespective of the presence or absence of clipped DCT. Therefore, in atherosclerotic lesions, phenotypic switching of VSMCs may facilitate pro-migratory effects of PDGF on VSMCs by suppressing posttranslational Ca V 1.2 modifications.

MicroRNA, miR-374b, directly targets Myf6 and negatively regulates C2C12 myoblasts differentiation

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Ma, Zhiyuan; Sun, Xiaorui; Xu, Dequan; Xiong, Yuanzhu; Zuo, Bo, E-mail: zuobo@mail.hzau.edu.cn

2015-11-27

Myogenesis is a complex process including myoblast proliferation, differentiation and myotube formation and is controlled by myogenic regulatory factors (MRFs), MyoD, MyoG, Myf5 and Myf6 (also known as MRF4). MicroRNA is a kind of ∼22 nt-long non-coding small RNAs, and act as key transcriptional or post-transcriptional regulators of gene expression. Identification of miRNAs involved in the regulation of muscle genes could improve our understanding of myogenesis process. In this study, we investigated the regulation of Myf6 gene by miRNAs. We showed that miR-374b specifically bound to the 3'untranslated region (UTR) of Myf6 and down-regulated the expression of Myf6 gene at both mRNA and protein level. Furthermore, miR-374b is ubiquitously expressed in the tissues of adult C57BL6 mouse, and the mRNA abundance increases first and then decreases during C2C12 myoblasts differentiation. Over-expression of miR-374b impaired C2C12 cell differentiation, while inhibiting miR-374b expression by 2′-O-methyl antisense oligonucleotides promoted C2C12 cell differentiation. Taken together, our findings identified miR-374b directly targets Myf6 and negatively regulates myogenesis. - Highlights: • MiR-374b directly targets 3′UTR of Myf6. • MiR-374b negatively regulates Myf6 in C2C12 cells. • MiR-374b abundance significiently changes during C2C12 cells differentiation. • MiR-374b negatively regulates C2C12 cells differentiation.

Cytoprotective Role of Nrf2 in Electrical Pulse Stimulated C2C12 Myotube.

Directory of Open Access Journals (Sweden)

Masaki Horie

Full Text Available Regular physical exercise is central to a healthy lifestyle. However, exercise-related muscle contraction can induce reactive oxygen species and reactive nitrogen species (ROS/RNS production in skeletal muscle. The nuclear factor-E2-related factor-2 (Nrf2 transcription factor is a cellular sensor for oxidative stress. Regulation of nuclear Nrf2 signaling regulates antioxidant responses and protects organ structure and function. However, the role of Nrf2 in exercise- or contraction-induced ROS/RNS production in skeletal muscle is not clear. In this study, using differentiated C2C12 cells and electrical pulse stimulation (EPS of muscle contraction, we explored whether Nrf2 plays a role in the skeletal muscle response to muscle contraction-induced ROS/RNS. We found that EPS (40 V, 1 Hz, 2 ms stimulated ROS/RNS accumulation and Nrf2 activation. We also showed that expression of NQO1, HO-1 and GCLM increased after EPS-induced muscle contraction and was remarkably suppressed in cells with Nrf2 knockdown. We also found that the antioxidant N-acetylcysteine (NAC significantly attenuated Nrf2 activation after EPS, whereas the nitric oxide synthetase inhibitor Nω-nitro-L-arginine methyl ester (L-NAME did not. Furthermore, Nrf2 knockdown after EPS markedly decreased ROS/RNS redox potential and cell viability and increased expression of the apoptosis marker Annexin V in C2C12 myotubes. These results indicate that Nrf2 activation and expression of Nrf2 regulated-genes protected muscle against the increased ROS caused by EPS-induced muscle contraction. Thus, our findings suggest that Nrf2 may be a key factor for preservation of muscle function during muscle contraction.

Increased radiation-induced transformation in C3H/10T1/2 cells after transfer of an exogenous c-myc gene

International Nuclear Information System (INIS)

Sorrentino, V.; Drozdoff, V.; Zeitz, L.; Fleissner, E.

1987-01-01

C3H/10T 1/2 cells were infected with a retroviral vector expressing a mouse c-myc oncogene and a drug-selection marker. The resulting cells, morphologically indistinguishable from C3H/10T l/1, displayed a greatly enhanced sensitivity to neoplastic transformation by ionizing radiation or by a chemical carcinogen. Constitutive expression of myc therefore appears to synergize with an initial carcinogenic event, providing a function analogous to a subsequent event that apparently is required for the neoplastic transformation of these cells. This cell system should prove useful in exploring early stages in radiation-induced transformation

Molecular resonances in sub-Coulomb energy region (12C-12C, 12C-24Mg, 12C-9Be systems)

International Nuclear Information System (INIS)

Takimoto, Kiyohiko; Shimomura, Susumu; Tanaka, Makoto; Murakami, Tetsuya; Fukada, Mamoru; Sakaguchi, Atsushi

1982-01-01

Molecular resonance in sub-Coulomb energy region was studied on 12 C- 12 C, 12 C- 24 Mg and 12 C- 9 Be systems. The excitation functions and the angular distributions were measured on the reactions 12 C( 12 C, 8 Besub(g,s,)) 16 Osub(g,s,), 24 Mg( 12 C, α) 32 S and 9 Be ( 12 C, 8 Besub(g,s,)) 13 Csub(g,s,). Sub-Coulomb resonances were observed in all systems and the contribution of the 12 Csub(2nd)*(0 + , 7.65 MeV) state is proposed. (author)

Screening effects on 12C+12C fusion reaction

Science.gov (United States)

Koyuncu, F.; Soylu, A.

2018-05-01

One of the important reactions for nucleosynthesis in the carbon burning phase in high-mass stars is the 12C+12C fusion reaction. In this study, we investigate the influences of the nuclear potentials and screening effect on astrophysically interesting 12C+12C fusion reaction observables at sub-barrier energies by using the microscopic α–α double folding cluster (DFC) potential and the proximity potential. In order to model the screening effects on the experimental data, a more general exponential cosine screened Coulomb (MGECSC) potential including Debye and quantum plasma cases has been considered in the calculations for the 12C+12C fusion reaction. In the calculations of the reaction observables, the semi-classical Wentzel-Kramers-Brillouin (WKB) approach and coupled channel (CC) formalism have been used. Moreover, in order to investigate how the potentials between 12C nuclei produce molecular cluster states of 24Mg, the normalized resonant energy states of 24Mg cluster bands have been calculated for the DFC potential. By analyzing the results produced from the fusion of 12C+12C, it is found that taking into account the screening effects in terms of MGECSC is important for explaining the 12C+12C fusion data, and the microscopic DFC potential is better than the proximity potential in explaining the experimental data, also considering that clustering is dominant for the structure of the 24Mg nucleus. Supported by the Turkish Science and Research Council (TÜBİTAK) with (117R015)

Retinoid acid-induced microRNA-27b-3p impairs C2C12 myoblast proliferation and differentiation by suppressing α-dystrobrevin

Energy Technology Data Exchange (ETDEWEB)

Li, Nan; Tang, Yi; Liu, Bo; Cong, Wei; Liu, Chao, E-mail: liuchao_19760711@yahoo.com; Xiao, Jing, E-mail: xiaoj@dmu.edu.cn

2017-01-15

We previously reported that excess retinoic acid (RA) resulted in hypoplastic and derangement of myofilaments in embryonic tongue by inhibiting myogenic proliferation and differentiation through CamKIID pathway. Our further studies revealed that the expression of a series of miRNAs was altered by RA administration in embryonic tongue as well as in C2C12 cells. Thus, if excess RA impairs myogenic proliferation and differentiation through miRNAs is taken into account. In present study, miR-27b-3p was found up-regulated in RA-treated C2C12 cells as in embryonic tongue, and predicted to target the 3′UTR of α-dystrobrevin (DTNA). Luciferase reporter assays confirmed the direct interaction between miR-27b-3p and the 3′UTR of DTNA. MiR-27b-3p mimics recapitulated the RA repression on DTNA expression, C2C12 proliferation and differentiation, while the miR-27b-3p inhibitor circumvented these defects resulting from excess RA. As expected, the effects of siDTNA on C2C12 were coincided with those by RA treatment or miR-27b-3p mimics. Therefore, these findings indicated that excess RA inhibited the myoblast proliferation and differentiation by up-regulating miR-27b-3p to target DTNA, which implied a new mechanism in myogenic hypoplasia. - Highlights: • A mechanism that RA results in tongue deformity by disrupting the myogenesis. • A non-muscle specific miR mediating the RA suppression on tongue myogenesis. • A target gene of non-muscle specific miR involved in RA induced tongue deformity.

Mechanical stimulation of C2C12 cells increases m-calpain expression, focal adhesion plaque protein degradation

DEFF Research Database (Denmark)

Grossi, Alberto; Karlsson, Anders H; Lawson, Moira Ann

2008-01-01

. Stimulation due to stretch- or load-induced signaling is now beginning to be understood as a factor which affects gene sequences, protein synthesis and an increase in Ca2+ influx in myocytes. Evidence of the involvement of Ca2+ -dependent activity in myoblast fusion, cell membrane and cytoskeleton component...... reorganization due to the activity of the ubiquitous proteolytic enzymes, calpains, has been reported. Whether there is a link between stretch- or load-induced signaling and calpain expression and activation is not known. Using a magnetic bead stimulation assay and C2C12 mouse myoblasts cell population, we have...... demonstrated that mechanical stimulation via laminin receptors leads to an increase in m-calpain expression, but no increase in the expression of other calpain isoforms. Our study revealed that after a short period of stimulation, m-calpain relocates into focal adhesion complexes and is followed by a breakdown...

Sodium arsenite represses the expression of myogenin in C2C12 mouse myoblast cells through histone modifications and altered expression of Ezh2, Glp, and Igf-1

Energy Technology Data Exchange (ETDEWEB)

Hong, Gia-Ming [Environmental Toxicology Graduate Program, Clemson University, 132 Long Hall, Clemson, SC 29634 (United States); Present address: The University of Chicago, Section of Hematology/Oncology, 900 E. 57th Street, Room 7134, Chicago, IL 60637 (United States); Bain, Lisa J., E-mail: lbain@clemson.edu [Environmental Toxicology Graduate Program, Clemson University, 132 Long Hall, Clemson, SC 29634 (United States); Department of Biological Sciences, Clemson University, 132 Long Hall, Clemson, SC 29634 (United States)

2012-05-01

Arsenic is a toxicant commonly found in water systems and chronic exposure can result in adverse developmental effects including increased neonatal death, stillbirths, and miscarriages, low birth weight, and altered locomotor activity. Previous studies indicate that 20 nM sodium arsenite exposure to C2C12 mouse myocyte cells delayed myoblast differentiation due to reduced myogenin expression, the transcription factor that differentiates myoblasts into myotubes. In this study, several mechanisms by which arsenic could alter myogenin expression were examined. Exposing differentiating C2C12 cells to 20 nM arsenic increased H3K9 dimethylation (H3K9me2) and H3K9 trimethylation (H3K9me3) by 3-fold near the transcription start site of myogenin, which is indicative of increased repressive marks, and reduced H3K9 acetylation (H3K9Ac) by 0.5-fold, indicative of reduced permissive marks. Protein expression of Glp or Ehmt1, a H3-K9 methyltransferase, was also increased by 1.6-fold in arsenic-exposed cells. In addition to the altered histone remodeling status on the myogenin promoter, protein and mRNA levels of Igf-1, a myogenic growth factor, were significantly repressed by arsenic exposure. Moreover, a 2-fold induction of Ezh2 expression, and an increased recruitment of Ezh2 (3.3-fold) and Dnmt3a (∼ 2-fold) to the myogenin promoter at the transcription start site (− 40 to + 42), were detected in the arsenic-treated cells. Together, we conclude that the repressed myogenin expression in arsenic-exposed C2C12 cells was likely due to a combination of reduced expression of Igf-1, enhanced nuclear expression and promoter recruitment of Ezh2, and altered histone remodeling status on myogenin promoter (− 40 to + 42). -- Highlights: ► Igf-1 expression is decreased in C2C12 cells after 20 nM arsenite exposure. ► Arsenic exposure alters histone remodeling on the myogenin promoter. ► Glp expression, a H3–K9 methyltransferase, was increased in arsenic-exposed cells. ► Ezh2

Comparison of heat and/or radiation sensitivity and membrane composition of seven X-ray-transformed C3H 10T1/2 cell lines and normal C3H 10T1/2 cells

International Nuclear Information System (INIS)

Raaphorst, G.P.; Vadasz, J.A.; Azzam, E.I.; Sargent, M.D.; Borsa, J.; Einspenner, M.

1985-01-01

C3H 10T1/2 mouse embryo cells were transformed by X-irradiation, and seven transformed clones were isolated and propagated as cell lines. Some of these cell lines produced tumors in syngeneic mice and grew in agarose while the normal C3H 10T1/2 cell line did not possess these characteristics. Exponentially growing cell cultures with comparable cell-cycle distributions as measured by flow cytometry were tested for heat and X-ray sensitivity. The heat and X-ray sensitivity varied randomly compared to the normal cell line. One cell line was more heat resistant and one more heat sensitive than the normal cell line, and the others had sensitivities comparable to the normal cell line. Measurements on some of the biochemical parameters of the particulate fraction of cells after sonication and 24,000 X g centrifugation showed that altered thermal sensitivity was not correlated with protein, cholesterol, or phospholipid content of this fraction

Curcumin enhances the mitomycin C-induced cytotoxicity via downregulation of MKK1/2-ERK1/2-mediated Rad51 expression in non-small cell lung cancer cells

International Nuclear Information System (INIS)

Ko, Jen-Chung; Tsai, Min-Shao; Weng, Shao-Hsing; Kuo, Ya-Hsun; Chiu, Yu-Fan; Lin, Yun-Wei

2011-01-01

Curcumin (diferuloylmethane), a major active component of turmeric (Curcuma longa), has been reported to suppress the proliferation of a wide variety of tumor cells. Rad51 is a key protein in the homologous recombination (HR) pathway of DNA double-strand break repair, and HR represents a novel target for cancer therapy. A high expression of Rad51 has been reported in chemo- or radio-resistant carcinomas. Therefore, in the current study, we will examine whether curcumin could enhance the effects of mitomycin C (MMC), a DNA interstrand cross-linking agent, to induce cytotoxicity by decreasing Rad51 expression. Exposure of two human non-small lung cancer (NSCLC) cell lines (A549 and H1975) to curcumin could suppress MMC-induced MKK1/2-ERK1/2 signal activation and Rad51 protein expression. Enhancement of ERK1/2 activation by constitutively active MKK1/2 (MKK1/2-CA) increased Rad51 protein levels in curcumin and MMC co-treated human lung cells. Moreover, the synergistic cytotoxic effect induced by curcumin combined with MMC was decreased by MKK1-CA-mediated enhancement of ERK1/2 activation by a significant degree. In contrast, MKK1/2 inhibitor, U0126 was shown to augment the cytotoxicity of curcumin and MMC through downregulation of ERK1/2 activation and Rad51 expression. Depletion of endogenous Rad51 expression by siRad51 RNA transfection significantly enhanced MMC and/or curcumin induced cell death and cell growth inhibition. In contrast, an overexpression of Rad51 protected lung cancer cells from synergistic cytotoxic effects induced by curcumin and MMC. We concluded that Rad51 inhibition may be an additional action mechanism for enhancing the chemosensitization of MMC by curcumin in NSCLC. - Highlights: → Curcumin downregulates MKK-ERK-mediated Rad51 expression. → Curcumin enhances mitomycin C-induced cytotoxicity. → Rad51 protects cells from cytotoxic effects induced by curcumin and mitomycin C. → Rad51 inhibition enhances the chemosensitization of

Expressional studies of the aldehyde oxidase (AOX1) gene during myogenic differentiation in C2C12 cells

Energy Technology Data Exchange (ETDEWEB)

Kamli, Majid Rasool; Kim, Jihoe; Pokharel, Smritee; Jan, Arif Tasleem [School of Biotechnology, Yeungnam University, Gyeongsan 712-749 (Korea, Republic of); Lee, Eun Ju [School of Biotechnology, Yeungnam University, Gyeongsan 712-749 (Korea, Republic of); Bovine Genome Resources Bank, Yeungnam University, Gyeongsan 712-749 (Korea, Republic of); Choi, Inho, E-mail: inhochoi@ynu.ac.kr [School of Biotechnology, Yeungnam University, Gyeongsan 712-749 (Korea, Republic of); Bovine Genome Resources Bank, Yeungnam University, Gyeongsan 712-749 (Korea, Republic of)

2014-08-08

Highlights: • AOX1 contributes to the formation of myotube. • Silencing of AOX1 reduces myotube formation. • AOX1 regulates MyoG gene expression. • AOX1 contributes to myogenesis via H{sub 2}O{sub 2}. - Abstract: Aldehyde oxidases (AOXs), which catalyze the hydroxylation of heterocycles and oxidation of a wide variety of aldehydic compounds, have been present throughout evolution from bacteria to humans. While humans have only a single functional aldehyde oxidase (AOX1) gene, rodents are endowed with four AOXs; AOX1 and three aldehyde oxidase homologs (AOH1, AOH2 and AOH3). In continuation of our previous study conducted to identify genes differentially expressed during myogenesis using a microarray approach, we investigated AOX1 with respect to its role in myogenesis to conceptualize how it is regulated in C2C12 cells. The results obtained were validated by silencing of the AOX1 gene. Analysis of their fusion index revealed that formation of myotubes showed a marked reduction of up to 40% in AOX1{sub kd} cells. Expression of myogenin (MYOG), one of the marker genes used to study myogenesis, was also found to be reduced in AOX1{sub kd} cells. AOX1 is an enzyme of pharmacological and toxicological importance that metabolizes numerous xenobiotics to their respective carboxylic acids. Hydrogen peroxide (H{sub 2}O{sub 2}) produced as a by-product in this reaction is considered to be involved as a part of the signaling mechanism during differentiation. An observed reduction in the level of H{sub 2}O{sub 2} among AOX1{sub kd} cells confirmed production of H{sub 2}O{sub 2} in the reaction catalyzed by AOX1. Taken together, these findings suggest that AOX1 acts as a contributor to the process of myogenesis by influencing the level of H{sub 2}O{sub 2}.

Effective myotube formation in human adipose tissue-derived stem cells expressing dystrophin and myosin heavy chain by cellular fusion with mouse C2C12 myoblasts

Energy Technology Data Exchange (ETDEWEB)

Eom, Young Woo [Cell Therapy and Tissue Engineering Center, Wonju College of Medicine, Yonsei Univ., Wonju (Korea, Republic of); Biomedical Research Institute, Lifeliver Co., Ltd., Suwon (Korea, Republic of); Lee, Jong Eun; Yang, Mal Sook; Jang, In Keun; Kim, Hyo Eun; Lee, Doo Hoon; Kim, Young Jin [Biomedical Research Institute, Lifeliver Co., Ltd., Suwon (Korea, Republic of); Park, Won Jin [Dr. Park' s Aesthetic Clinic, Seoul (Korea, Republic of); Kong, Jee Hyun; Shim, Kwang Yong [Department of Hematology-Oncology, Wonju College of Medicine, Yonsei Univ., Wonju (Korea, Republic of); Lee, Jong In, E-mail: oncochem@yonsei.ac.kr [Department of Hematology-Oncology, Wonju College of Medicine, Yonsei Univ., Wonju (Korea, Republic of); Kim, Hyun Soo, E-mail: khsmd@unitel.co.kr [Department of Hematology-Oncology, Wonju College of Medicine, Yonsei Univ., Wonju (Korea, Republic of)

2011-04-29

Highlights: {yields} hASCs were differentiated into skeletal muscle cells by treatment with 5-azacytidine, FGF-2, and the supernatant of cultured hASCs. {yields} Dystrophin and MyHC were expressed in late differentiation step by treatment with the supernatant of cultured hASCs. {yields} hASCs expressing dystrophin and MyHC contributed to myotube formation during co-culture with mouse myoblast C2C12 cells. -- Abstract: Stem cell therapy for muscular dystrophies requires stem cells that are able to participate in the formation of new muscle fibers. However, the differentiation steps that are the most critical for this process are not clear. We investigated the myogenic phases of human adipose tissue-derived stem cells (hASCs) step by step and the capability of myotube formation according to the differentiation phase by cellular fusion with mouse myoblast C2C12 cells. In hASCs treated with 5-azacytidine and fibroblast growth factor-2 (FGF-2) for 1 day, the early differentiation step to express MyoD and myogenin was induced by FGF-2 treatment for 6 days. Dystrophin and myosin heavy chain (MyHC) expression was induced by hASC conditioned medium in the late differentiation step. Myotubes were observed only in hASCs undergoing the late differentiation step by cellular fusion with C2C12 cells. In contrast, hASCs that were normal or in the early stage were not involved in myotube formation. Our results indicate that stem cells expressing dystrophin and MyHC are more suitable for myotube formation by co-culture with myoblasts than normal or early differentiated stem cells expressing MyoD and myogenin.

Effective myotube formation in human adipose tissue-derived stem cells expressing dystrophin and myosin heavy chain by cellular fusion with mouse C2C12 myoblasts

International Nuclear Information System (INIS)

Eom, Young Woo; Lee, Jong Eun; Yang, Mal Sook; Jang, In Keun; Kim, Hyo Eun; Lee, Doo Hoon; Kim, Young Jin; Park, Won Jin; Kong, Jee Hyun; Shim, Kwang Yong; Lee, Jong In; Kim, Hyun Soo

2011-01-01

Highlights: → hASCs were differentiated into skeletal muscle cells by treatment with 5-azacytidine, FGF-2, and the supernatant of cultured hASCs. → Dystrophin and MyHC were expressed in late differentiation step by treatment with the supernatant of cultured hASCs. → hASCs expressing dystrophin and MyHC contributed to myotube formation during co-culture with mouse myoblast C2C12 cells. -- Abstract: Stem cell therapy for muscular dystrophies requires stem cells that are able to participate in the formation of new muscle fibers. However, the differentiation steps that are the most critical for this process are not clear. We investigated the myogenic phases of human adipose tissue-derived stem cells (hASCs) step by step and the capability of myotube formation according to the differentiation phase by cellular fusion with mouse myoblast C2C12 cells. In hASCs treated with 5-azacytidine and fibroblast growth factor-2 (FGF-2) for 1 day, the early differentiation step to express MyoD and myogenin was induced by FGF-2 treatment for 6 days. Dystrophin and myosin heavy chain (MyHC) expression was induced by hASC conditioned medium in the late differentiation step. Myotubes were observed only in hASCs undergoing the late differentiation step by cellular fusion with C2C12 cells. In contrast, hASCs that were normal or in the early stage were not involved in myotube formation. Our results indicate that stem cells expressing dystrophin and MyHC are more suitable for myotube formation by co-culture with myoblasts than normal or early differentiated stem cells expressing MyoD and myogenin.

Sodium arsenite alters cell cycle and MTHFR, MT1/2, and c-Myc protein levels in MCF-7 cells

International Nuclear Information System (INIS)

Ruiz-Ramos, Ruben; Lopez-Carrillo, Lizbeth; Albores, Arnulfo; Hernandez-Ramirez, Raul U.; Cebrian, Mariano E.

2009-01-01

There is limited available information on the effects of arsenic on enzymes participating in the folate cycle. Therefore, our aim was to evaluate the effects of sodium arsenite on the protein levels of methylenetetrahydrofolate reductase (MTHFR) and dihydrofolate reductase (DHFR) and its further relationship with the expression MT1/2 and c-myc in MCF-7 cells. Arsenite treatment (0-10 μM) for 4 h decreased MTHFR levels in a concentration-dependent fashion without significant effects on DHFR. The effects on MTHFR were observed at arsenite concentrations not significantly affecting cell viability. We also observed an increase in S-phase recruitment at all concentrations probed. Lower concentrations (< 5 μM) induced cell proliferation, showing a high proportion of BrdU-stained cells, indicating a higher DNA synthesis rate. However, higher concentrations (≥ 5 μM) or longer treatment periods induced apoptosis. Arsenite also induced dose-dependent increases in MT1/2 and c-Myc protein levels. The levels of MTHFR were inversely correlated to MT1/2 and c-Myc overexpression and increased S-phase recruitment. Our findings indicate that breast epithelial cells are responsive to arsenite and suggest that exposure may pose a risk for breast cancer. The reductions in MTHFR protein levels contribute to understand the mechanisms underlying the induction of genes influencing growth regulation, such as c-myc and MT1/2. However, further research is needed to ascertain if the effects here reported following short-time and high-dose exposure are relevant for human populations chronically exposed to low arsenic concentrations.

Functional aspects of dexamethasone upregulated nicotinic acetylcholine receptors in C2C12 myotubes

NARCIS (Netherlands)

Maestrone, E; Lagostena, L; Henning, RH; DenHertog, A; Nobile, M

Three days of treatment with the glucocorticoid dexamethasone (1 nM-mu M) induced a concentration-dependent up-regulation of muscle nicotinic acetylcholine receptor (nAChR) in C2C12 mouse myotubes (EC(50)=10+/-7.3 nM), as assessed by [H-3]alpha-BuTx binding. The maximum increase in binding amounted

Distal C terminus of CaV1.2 channels plays a crucial role in the neural differentiation of dental pulp stem cells.

Directory of Open Access Journals (Sweden)

Jianping Ge

Full Text Available L-type voltage-dependent CaV1.2 channels play an important role in the maintenance of intracellular calcium homeostasis, and influence multiple cellular processes. C-terminal cleavage of CaV1.2 channels was reported in several types of excitable cells, but its expression and possible roles in non-excitable cells is still not clear. The aim of this study was to determine whether distal C-terminal fragment of CaV1.2 channels is present in rat dental pulp stem cells and its possible role in the neural differentiation of rat dental pulp stem cells. We generated stable CaV1.2 knockdown cells via short hairpin RNA (shRNA. Rat dental pulp stem cells with deleted distal C-terminal of CaV1.2 channels lost the potential of differentiation to neural cells. Re-expression of distal C-terminal of CaV1.2 rescued the effect of knocking down the endogenous CaV1.2 on the neural differentiation of rat dental pulp stem cells, indicating that the distal C-terminal of CaV1.2 is required for neural differentiation of rat dental pulp stem cells. These results provide new insights into the role of voltage-gated Ca(2+ channels in stem cells during differentiation.

C2-Ceramide Induces Cell Death and Protective Autophagy in Head and Neck Squamous Cell Carcinoma Cells

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Wenyuan Zhu

2014-02-01

Full Text Available Ceramides are second messengers involved in several intracellular processes in cancer cells, amongst others. The aim of this study was to evaluate the anti-tumor efficacy of C2-ceramide (C2-Cer; N-acetyl-D-sphingosine by investigating cell death and autophagy in head and neck squamous cell carcinoma (HNSCC cells. C2-Cer showed concentration-dependent cytotoxicity in HN4 and HN30 cell lines. It simultaneously induced caspase-3-independent apoptosis and programmed necrosis. C2-Cer markedly increased the expression level of microtubule-associated protein 1 light chain 3B (LC3B type II associated with protective autophagy. An autophagy inhibitor enhanced C2-Cer-mediated cytotoxicity, while a programmed-necrosis inhibitor produced the opposite effect. Furthermore, C2-Cer up-regulated the phosphorylation of extracellular signal-regulated kinase 1/2, but down-regulated its downstream substrate phospho-mammalian target of rapamycin (p-mTOR during the autophagy process. These results suggested that C2-Cer exerts anti-tumor effects by inducing programmed apoptosis and necrosis in HNSCC, and these cytotoxic effects are enhanced by an autophagy inhibitor.

Microscopic investigation of the 12C + 12C interaction

International Nuclear Information System (INIS)

Baye, D.; Pecher, N.; Brussels Univ.

1982-01-01

The 12 C + 12 C system is studied in the framework of the generator coordinate method. Each 12 C nucleus is described by a closed psub(3/2) subshell. Phase shifts and resonances are determined for several effective two-body interactions involving a spin-orbit term. The existence and properties of simple local equivalent potentials for the 12 C + 12 C collision are discussed. The 12 C + 12 C system is too light to be well described by potentials independent of the angular momentum or weakly dependent on it. (orig.)

The transcription factors CREB and c-Fos play key roles in NCAM-mediated neuritogenesis in PC12-E2 cells

DEFF Research Database (Denmark)

Jessen, U; Novitskaya, V; Pedersen, N

2001-01-01

The neural cell adhesion molecule (NCAM) stimulates axonal outgrowth by activation of the Ras-mitogen activated protein kinase (MAPK) pathway and by generation of arachidonic acid. We investigated whether the transcription factors, cyclic-AMP response-element binding protein (CREB) and c-Fos play...... roles in this process by estimating NCAM-dependent neurite outgrowth from PC12-E2 cells grown in co-culture with NCAM-negative or NCAM-positive fibroblasts. PC12-E2 cells were transiently transfected with expression plasmids encoding wild-type or dominant negative forms of CREB and c-Fos or an activated...... form of the MAPK kinase, MEK2. Alternatively, PC12-E2 cells were treated with arachidonic acid, the cAMP analogue dBcAMP, or protein kinase A (PKA) inhibitors. The negative forms of CREB and c-Fos inhibited neurite outgrowth mediated by NCAM, arachidonic acid, dBcAMP, or MEK2. Neither CREB nor c...

THE {sup 12}C + {sup 12}C REACTION AND THE IMPACT ON NUCLEOSYNTHESIS IN MASSIVE STARS

Energy Technology Data Exchange (ETDEWEB)

Pignatari, M. [Department of Physics, University of Basel, Klingelbergstrasse 82, CH-4056 Basel (Switzerland); Hirschi, R.; Bennett, M. [Astrophysics Group, EPSAM Institute, Keele University, Keele, ST5 5BG (United Kingdom); Wiescher, M.; Beard, M. [Department of Physics, University of Notre Dame, Notre Dame, IN 46556 (United States); Gallino, R. [Universita' di Torino, Torino, Via Pietro Giuria 1, I-10126 Torino (Italy); Fryer, C.; Rockefeller, G. [Computational Physics and Methods (CCS-2), LANL, Los Alamos, NM 87545 (United States); Herwig, F.; Timmes, F. X., E-mail: marco.pignatari@unibas.ch [The Joint Institute for Nuclear Astrophysics, Notre Dame, IN 46556 (United States)

2013-01-01

Despite much effort in the past decades, the C-burning reaction rate is uncertain by several orders of magnitude, and the relative strength between the different channels {sup 12}C({sup 12}C, {alpha}){sup 20}Ne, {sup 12}C({sup 12}C, p){sup 23}Na, and {sup 12}C({sup 12}C, n){sup 23}Mg is poorly determined. Additionally, in C-burning conditions a high {sup 12}C+{sup 12}C rate may lead to lower central C-burning temperatures and to {sup 13}C({alpha}, n){sup 16}O emerging as a more dominant neutron source than {sup 22}Ne({alpha}, n){sup 25}Mg, increasing significantly the s-process production. This is due to the chain {sup 12}C(p, {gamma}){sup 13}N followed by {sup 13}N({beta} +){sup 13}C, where the photodisintegration reverse channel {sup 13}N({gamma}, p){sup 12}C is strongly decreasing with increasing temperature. Presented here is the impact of the {sup 12}C+{sup 12}C reaction uncertainties on the s-process and on explosive p-process nucleosynthesis in massive stars, including also fast rotating massive stars at low metallicity. Using various {sup 12}C+{sup 12}C rates, in particular an upper and lower rate limit of {approx}50,000 higher and {approx}20 lower than the standard rate at 5 Multiplication-Sign 10{sup 8} K, five 25 M {sub Sun} stellar models are calculated. The enhanced s-process signature due to {sup 13}C({alpha}, n){sup 16}O activation is considered, taking into account the impact of the uncertainty of all three C-burning reaction branches. Consequently, we show that the p-process abundances have an average production factor increased up to about a factor of eight compared with the standard case, efficiently producing the elusive Mo and Ru proton-rich isotopes. We also show that an s-process being driven by {sup 13}C({alpha}, n){sup 16}O is a secondary process, even though the abundance of {sup 13}C does not depend on the initial metal content. Finally, implications for the Sr-peak elements inventory in the solar system and at low metallicity are

Synthetic Swan band profile of (1,0) of 12C12C and (0,0) of 12C12C and 12C13C in comets

International Nuclear Information System (INIS)

Swamy, K.S.K.

1987-01-01

The statistical equilibrium calculations of the 12 C 13 C molecule based on the resonance fluorescence process give similar results to those of the normal molecule. Therefore the assumption that the observed intensities of bands of the normal and the isotopic molecule differ only by their abundance ratio is reasonable. The synthetic profile of the (1,0) Swan band of 12 C 13 C (0,0) band of 12 C 12 C and 12 C 13 C have been calculated. The relative merits of using the rotational structure of the (1,0) or (0,0) band for the determination of the isotopic ratio 12 C/ 13 C is discussed briefly. (author)

[TRPM8 mediates PC-12 neuronal cell apoptosis induced by oxygen-glucose deprivation through cAMP-PKA/UCP4 signaling].

Science.gov (United States)

Li, Hong-Wei; Zhou, Bin; Zhang, Hai-Hong

2016-08-20

To explore the molecular mechanism responsible for apoptosis of PC-12 neuronal cells induced by oxygen-glucose deprivation (OGD). PC12 cells were exposed to OGD for 24 h to simulate ischemia-reperfusion injury. Flow cytometry was employed detect the cell apoptosis, and the expresions of TRPM8, UCP4, cAMP and PKA in the exposed cells were detected with RT-PCR and Western blotting. The changes in the expressions of Bax, Bcl-2, cAMP, PKA and UCP4 proteins were detected in the exposed cells in resposne to inhibition of TRPM8 and cAMP-PKA signal or over-expression of UCP4. OGD for 24 induced obvious apoptosis in PC-12 cells and caused TRPM8 over-expression and inhibition of UCP4 and cAMP-PKA signaling. Inhibiting TRPM8 expression reduced the cell apoptosis and up-regulated cAMP, p-PKA and UCP4 in the cells exposed to OGD. In cells exposed to OGD, inhibition of TRPM8 and cAMP-PKA signaling suppressed the expressio of UCP4 and increased the cell apoptosis. TRPM8 mediates OGD-induced PC12 cell apoptosis through cAMP-PKA/UCP4 signaling.

Measurements of Ay(θ) for 12C(n,n)12C from En=2.2 to 8.5 MeV

International Nuclear Information System (INIS)

Roper, C.D.; Tornow, W.; Braun, R.T.; Chen, Q.; Crowell, A.; Trotter, D. Gonzalez; Howell, C.R.; Salinas, F.; Setze, R.; Walter, R.L.; Chen Zemin; Tang Hongqing; Zhou Zuying

2005-01-01

The analyzing power A y (θ) for neutron elastic scattering from 12 C has been measured for 33 neutron energies between E n =2.2 and 8.5 MeV in the angular range from 25 deg. to 145 deg. in the laboratory system. The primary motivation for these measurements is the need for an accurate knowledge of A y (θ) for 12 C(n,n) 12 C elastic scattering to enable corrections to high-precision neutron-proton and neutron-deuteron A y (θ) data in the neutron-energy range below E n =30 MeV. In their own right, 12 C(n,n) 12 C A y (θ) data are of crucial importance for improving both the parametrization of n- 12 C scattering and our knowledge of the level scheme of 13 C. The present A y (θ) data are compared with published data and previous phase-shift-analysis results

Investigation of low spin resonances in the 12C+12C and 16O+12C systems

International Nuclear Information System (INIS)

Uzureau, J.L.; Fieni, J.M.; Cocu, F.; Michaudon, A.

1979-01-01

The excitation functions of the 12 C( 12 C,α) 20 Ne and 12 C( 16 O,α) 24 Mg reactions were measured at different angles below the Coulomb barrier. Resonant structures were found at Esub(cm)=5.80 MeV in the 12 C+ 12 C system and at Esub(cm)=6.10 MeV in the 16 O+ 12 C system. From the analysis of angular distributions measured at the previous resonant energies, values of respectively Jsup(π)=0 + and 2 + have been deduced [fr

A fractionation method to identify qauntitative changes in protein expression mediated by IGF-1 on the proteome of murine C2C12 myoblasts

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Friedmann Theodore

2009-08-01

Full Text Available Abstract Although much is known about signal transduction downstream of insulin-like growth factor-1 (IGF-1, relatively little is known about the global changes in protein expression induced by this hormone. In this study, the acute effects of IGF-1 on the proteome of murine C2C12 cells were examined. Cells were treated with IGF-1 for up to 24 hours, lysed, and fractionated into cytosolic, nuclear, and insoluble portions. Proteins from the cytosolic fraction were further separated using a new batch ion-exchange chromatography method to reduce sample complexity, followed by two-dimensional (2D electrophoresis, and identification of selected proteins by mass spectrometry. PDQuest software was utilized to identify and catalogue temporal changes in protein expression during IGF-1 stimulation. In response to IGF-1 stimulation, expression of 23 proteins increased at least three-fold and expression of 17 proteins decreased at least three-fold compared with control un-stimulated C2C12 cells. Changes in expression of selected proteins from each group, including Rho-GDI, cofillin, RAD50, enolase, IκB kinase b (IκBKb and Hsp70 were confirmed by Western blotting. Additionally, the position of 136 'landmark' proteins whose expression levels and physicochemical properties did not change appreciably or consistently during IGF-1 treatment were mapped and identified. This characterization of large-scale changes in protein expression in response to growth factor stimulation of C2C12 cells will further help to establish a comprehensive understanding of the networks and pathways involved in the action of IGF-1.

Influence of the PDE5 inhibitor tadalafil on redox status and antioxidant defense system in C2C12 skeletal muscle cells.

Science.gov (United States)

Duranti, Guglielmo; Ceci, Roberta; Sgrò, Paolo; Sabatini, Stefania; Di Luigi, Luigi

2017-05-01

Phosphodiesterase type 5 inhibitors (PDE5Is), widely known for their beneficial effects onto male erectile dysfunction, seem to exert favorable effects onto metabolism as well. Tadalafil exposure increases oxidative metabolism of C2C12 skeletal muscle cells. A rise in fatty acid (FA) metabolism, requiring more oxygen, could induce a larger reactive oxygen species (ROS) release as a byproduct thus leading to a redox imbalance. The aim of this study was to determine how PDE5I tadalafil influences redox status in skeletal muscle cells to match the increasing oxidative metabolism. To this purpose, differentiated C2C12 skeletal muscle cells were treated with tadalafil and analyzed for total antioxidant capacity (TAC) and glutathione levels as marker of redox status; enzyme activity of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) engaged in antioxidant defense; and lipid peroxidation (TBARS) and protein carbonyls (PrCar) as markers of oxidative damage. Tadalafil increased total intracellular glutathione (tGSH), CAT, SOD, and GPx enzymatic activities while no changes were found in TAC. A perturbation of redox status, as showed by the decrease in the ratio between reduced/oxidized glutathione (GSH/GSSG), was observed. Nevertheless, it did not cause any change in TBARS and PrCar levels probably due to the enhancement in the antioxidant enzymatic network. Taken together, these data indicate that tadalafil, besides improving oxidative metabolism, may be beneficial to skeletal muscle cells by enhancing the enzymatic antioxidant system capacity.

Toxic mechanisms of 3-monochloropropane-1,2-diol on progesterone production in R2C rat leydig cells.

Science.gov (United States)

Sun, Jianxia; Bai, Shun; Bai, Weibin; Zou, Feiyan; Zhang, Lei; Su, Zhijian; Zhang, Qihao; Ou, Shiyi; Huang, Yadong

2013-10-16

3-Monochloropropane-1,2-diol (3-MCPD) is a well-known food processing contaminant that has been shown to impede the male reproductive function. However, its mechanism of action remains to be elucidated. In this study, the effects of 3-MCPD on progesterone production were investigated using R2C Leydig cells. 3-MCPD caused concentration-dependent inhibition of cell viability at the IC25, IC50, and IC75 levels of 1.027, 1.802, and 3.160 mM, respectively. Single cell gel/comet assay and atomic force microscopy assay showed that 3-MCPD significantly induced early apoptosis. In addition, 3-MCPD significantly reduced progesterone production by reducing the expression of cytochrome P450 side-chain cleavage enzyme, steroidogenic acute regulatory protein, and 3β-hydroxysteroid dehydrogenase in R2C cells. The change in steroidogenic acute regulatory protein expression was highly consistent with progesterone production. Furthermore, the mitochondrial membrane potential and cAMP significantly decreased.

Leucine-induced activation of translational initiation is partly regulated by the branched-chain α-keto acid dehydrogenase complex in C2C12 cells

International Nuclear Information System (INIS)

Nakai, Naoya; Shimomura, Yoshiharu; Tamura, Tomohiro; Tamura, Noriko; Hamada, Koichiro; Kawano, Fuminori; Ohira, Yoshinobu

2006-01-01

Branched-chain amino acid leucine has been shown to activate the translational regulators through the mammalian target of rapamycin. However, the leucine's effects are self-limiting because leucine promotes its own disposal by an oxidative pathway. The irreversible and rate-limiting step in the leucine oxidation pathway is catalyzed by the branched-chain α-keto acid dehydrogenase (BCKDH) complex. The complex contains E1 (α2β2), E2, and E3 subunits, and its activity is abolished by phosphorylation of the E1α subunit by BCKDH kinase. The relationship between the activity of BCKDH complex and leucine-mediated activation of the protein translation was investigated using the technique of RNA interference. The activity of BCKDH complex in C2C12 cell was modulated by transfection of small interfering RNA (siRNA) for BCKDH E2 subunit or BCKDH kinase. Transfection of siRNAs decreased the mRNA expression and protein amount of corresponding gene. Suppression of either E2 subunit or kinase produced opposite effects on the cell proliferation and the activation of translational regulators by leucine. Suppression of BCKDH kinase for 48 h resulted in decreasing cell proliferation. In contrast, E2 suppression led to increased amount of total cellular protein. The phosphorylation of p70 S6 kinase by leucine was increased in E2-siRNA transfected C2C12 cells, whereas the leucine's effect was diminished in kinase-siRNA transfected cells. These results suggest that the activation of the translational regulators by leucine was partly regulated by the activity of BCKDH complex

Comparison of aggregation behaviors between ionic liquid-type imidazolium gemini surfactant [C12-4-C12im]Br2 and its monomer [C12mim]Br on silicon wafer.

Science.gov (United States)

Ao, Mingqi; Xu, Guiying; Pang, Jinyu; Zhao, Taotao

2009-09-01

The aggregation of ionic liquid-type imidazolium gemini surfactant [C(12)-4-C(12)im]Br(2) on silicon wafer, which is compared with its monomer [C(12)mim]Br, have been studied. AFM morphology images and contact angle measurements suggest that the aggregations of [C(12)-4-C(12)im]Br(2) and [C(12)mim]Br on silicon wafer follow different mechanisms. Below the critical surface aggregation concentrations (CSAC), both surfactant molecules are adsorbed with their hydrophobic tails facing the air. But above the CSAC, [C(12)-4-C(12)im]Br(2) molecules finally form a bilayer structure with hydrophilic head groups facing the air, whereas [C(12)mim]Br molecules form a multilayer structure, and with increasing its concentration, the layer numbers increase with the hydrophobic chains and hydrophilic head groups facing the air by turns. Besides, the watery wettability of [C(12)-4-C(12)im]Br(2)-treated silica surface is lower than that of [C(12)mim]Br at the concentration of 5.0 cmc, and the infrared spectroscopy suggests that the poorer watery wettability of [C(12)-4-C(12)im]Br(2) may be relative to the less-ordered packing of methylene chains inside the aggregate. These different aggregation behaviors for the two surfactants ascribe to the different molecular structures and electrostatic interactions. This work would have certain theoretical guidance meaning on the modification of solid surface.

Leucine Modulates Mitochondrial Biogenesis and SIRT1-AMPK Signaling in C2C12 Myotubes

Directory of Open Access Journals (Sweden)

Chunzi Liang

2014-01-01

Full Text Available Previous studies from this laboratory demonstrate that dietary leucine protects against high fat diet-induced mitochondrial impairments and stimulates mitochondrial biogenesis and energy partitioning from adipocytes to muscle cells through SIRT1-mediated mechanisms. Moreover, β-hydroxy-β-methyl butyrate (HMB, a metabolite of leucine, has been reported to activate AMPK synergistically with resveratrol in C2C12 myotubes. Therefore, we hypothesize that leucine-induced activation of SIRT1 and AMPK is the central event that links the upregulated mitochondrial biogenesis and fatty acid oxidation in skeletal muscle. Thus, C2C12 myotubes were treated with leucine (0.5 mM, alanine (0.5 mM, valine (0.5 mM, EX527 (SIRT1 inhibitor, 25 μM, and Compound C (AMPK inhibitor, 25 μM alone or in combination to determine the roles of AMPK and SIRT1 in leucine-modulation of energy metabolism. Leucine significantly increased mitochondrial content, mitochondrial biogenesis-related genes expression, fatty acid oxidation, SIRT1 activity and gene expression, and AMPK phosphorylation in C2C12 myotubes compared to the controls, while EX527 and Compound C markedly attenuated these effects. Furthermore, leucine treatment for 24 hours resulted in time-dependent increases in cellular NAD+, SIRT1 activity, and p-AMPK level, with SIRT1 activation preceding that of AMPK, indicating that leucine activation of SIRT1, rather than AMPK, is the primary event.

File list: InP.Myo.20.AllAg.C2C12 [Chip-atlas[Archive

Lifescience Database Archive (English)

Full Text Available InP.Myo.20.AllAg.C2C12 mm9 Input control Muscle C2C12 SRX262224,SRX262225,SRX148229...695944 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/InP.Myo.20.AllAg.C2C12.bed ...

Excitation functions of the systems 12C+14C and 13C+12C

International Nuclear Information System (INIS)

Haindl, E.

1975-01-01

The excitation functions of the systems 12 C+ 14 C and 13 C+ 12 C are investigated for different exit channels. The excitation functions measured do not show correlated structures as in the system 12 C+ 12 C. (WL/AK) [de

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.

Pannexin channels mediate the acquisition of myogenic commitment in C2C12 reserve cells promoted by P2 receptor activation

Science.gov (United States)

Riquelme, Manuel A.; Cea, Luis A.; Vega, José L.; Puebla, Carlos; Vargas, Aníbal A.; Shoji, Kenji F.; Subiabre, Mario; Sáez, Juan C.

2015-01-01

The acquisition of myoblast commitment to the myogenic linage requires rises in intracellular free Ca2+ concentration ([Ca2+]i). Putative cell membrane pathways involved in these [Ca2+]i increments are P2 receptors (P2Rs) as well as connexin (Cx) and/or pannexin (Panx) hemichannels and channels (Cx HChs and Panx Chs), respectively, which are known to permeate Ca2+. Reserve cells (RCs) are uncommitted myoblasts obtained from differentiated C2C12 cell cultures, which acquire commitment upon replating. Regarding these cells, we found that extracellular ATP increases the [Ca2+]i via P2Rs. Moreover, ATP increases the plasma membrane permeability to small molecules and a non-selective membrane current, both of which were inhibited by Cx HCh/Panx1Ch blockers. However, RCs exposed to divalent cation-free saline solution, which is known to activate Cx HChs (but not Panx Chs), did not enhance membrane permeability, thus ruling out the possible involvement of Cx HChs. Moreover, ATP-induced membrane permeability was inhibited with blockers of P2Rs that activate Panx Chs. In addition, exogenous ATP induced the expression of myogenic commitment and increased MyoD levels, which was prevented by the inhibition of P2Rs or knockdown of Panx1 Chs. Similarly, increases in MyoD levels induced by ATP released by RCs were inhibited by Panx Ch/Cx HCh blockers. Myogenic commitment acquisition thus requires a feed-forward mechanism mediated by extracellular ATP, P2Rs, and Panx Chs. PMID:26000275

Hydrothermal studies in a new diamond anvil cell up to 10 GPa and from -190°C to 1200°C

Science.gov (United States)

Bassett, W. A.; Shen, A. H.; Bucknum, M.; Chou, I.-Ming

1993-06-01

The new hydrothermal diamond anvil cell (HDAC) has been designed for optical microscopy and X-ray diffraction at pressures up to 10 GPa and temperatures between -190°C and 1200°C. Laser light reffected from the top and bottom anvil faces and the top and bottom solid sample faces produce interference fringes that provide a very sensitive means of monitoring the volume of sample chamber and for observing volume and refractive index changes in solid samples due to transitions and reactions. Synchrotron radiation has been used to make X-ray diffraction patterns of samples under hydrothermal conditions. Individual heaters and individual thermocouples provide temperature control with an accuracy of ±0.5°C. Liquid nitrogen directly introduced into the HDAC has been used to reduce the sample temperature to -190°C. The α-β phase boundary of quartz has been used to calculate the transition pressures from measured transition temperatures. With this method we have redetermined 5 isochores of H2O up to 850°C and 1.2 GPa at which the solution rate of the quartz became so rapid that the quartz dissolved completely before the α-β transition could be observed. When silica solutions were cooled, opal spherules and rods formed.

Synthesis of 14C analogue of 1,2-diaryl-[2-14C]-pyrroles

International Nuclear Information System (INIS)

Saemian, N.; Shirvani, G.; Matloubi, H.

2007-01-01

Three 1,2-diaryl pyrroles selective COX-2 inhibitors, 2-(4-fluorophenyl)-5-methyl-1-(4-methylsulfonyl-phenyl)-1H pyrrole, 2-(4-fluorophenyl)-1- [4-(methylsulfonyl) phenyl]-1H-pyrrole and 4-[2-(4-fluorophenyl)-1H-pyrrol-1-yl]benzenesulfon-amide, all three labeled with 14 C in the 2-position were prepared from para-fluoro-benzaldehyde-[carbonyl- 14 C]. (author)

FOXO1 and GSK-3β Are Main Targets of Insulin-Mediated Myogenesis in C2C12 Muscle Cells.

Science.gov (United States)

Litwiniuk, Anna; Pijet, Barbara; Pijet-Kucicka, Maja; Gajewska, Małgorzata; Pająk, Beata; Orzechowski, Arkadiusz

2016-01-01

Myogenesis and muscle hypertrophy account for muscle growth and adaptation to work overload, respectively. In adults, insulin and insulin-like growth factor 1 stimulate muscle growth, although their links with cellular energy homeostasis are not fully explained. Insulin plays critical role in the control of mitochondrial activity in skeletal muscle cells, and mitochondria are essential for insulin action. The aim of this study was to elucidate molecular mechanism(s) involved in mitochondrial control of insulin-dependent myogenesis. The effects of several metabolic inhibitors (LY294002, PD98059, SB216763, LiCl, rotenone, oligomycin) on the differentiation of C2C12 myoblasts in culture were examined in the short-term (hours) and long-term (days) experiments. Muscle cell viability and mitogenicity were monitored and confronted with the activities of selected genes and proteins expression. These indices focus on the roles of insulin, glycogen synthase kinase 3 beta (GSK-3β) and forkhead box protein O1 (FOXO1) on myogenesis using a combination of treatments and inhibitors. Long-term insulin (10 nM) treatment in "normoglycemic" conditions led to increased myogenin expression and accelerated myogenesis in C2C12 cells. Insulin-dependent myogenesis was accompanied by the rise of mtTFA, MtSSB, Mfn2, and mitochondrially encoded Cox-1 gene expressions and elevated levels of proteins which control functions of mitochondria (kinase--PKB/AKT, mitofusin 2 protein--Mfn-2). Insulin, via the phosphatidylinositol 3-kinase (PI3-K)/AKT-dependent pathway reduced transcription factor FOXO1 activity and altered GSK-3β phosphorylation status. Once FOXO1 and GSK-3β activities were inhibited the rise in Cox-1 gene action and nuclear encoded cytochrome c oxidase subunit IV (COX IV) expressions were observed, even though some mRNA and protein results varied. In contrast to SB216763, LiCl markedly elevated Mfn2 and COX IV protein expression levels when given together with insulin. Thus

FOXO1 and GSK-3β Are Main Targets of Insulin-Mediated Myogenesis in C2C12 Muscle Cells.

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Anna Litwiniuk

Full Text Available Myogenesis and muscle hypertrophy account for muscle growth and adaptation to work overload, respectively. In adults, insulin and insulin-like growth factor 1 stimulate muscle growth, although their links with cellular energy homeostasis are not fully explained. Insulin plays critical role in the control of mitochondrial activity in skeletal muscle cells, and mitochondria are essential for insulin action. The aim of this study was to elucidate molecular mechanism(s involved in mitochondrial control of insulin-dependent myogenesis. The effects of several metabolic inhibitors (LY294002, PD98059, SB216763, LiCl, rotenone, oligomycin on the differentiation of C2C12 myoblasts in culture were examined in the short-term (hours and long-term (days experiments. Muscle cell viability and mitogenicity were monitored and confronted with the activities of selected genes and proteins expression. These indices focus on the roles of insulin, glycogen synthase kinase 3 beta (GSK-3β and forkhead box protein O1 (FOXO1 on myogenesis using a combination of treatments and inhibitors. Long-term insulin (10 nM treatment in "normoglycemic" conditions led to increased myogenin expression and accelerated myogenesis in C2C12 cells. Insulin-dependent myogenesis was accompanied by the rise of mtTFA, MtSSB, Mfn2, and mitochondrially encoded Cox-1 gene expressions and elevated levels of proteins which control functions of mitochondria (kinase--PKB/AKT, mitofusin 2 protein--Mfn-2. Insulin, via the phosphatidylinositol 3-kinase (PI3-K/AKT-dependent pathway reduced transcription factor FOXO1 activity and altered GSK-3β phosphorylation status. Once FOXO1 and GSK-3β activities were inhibited the rise in Cox-1 gene action and nuclear encoded cytochrome c oxidase subunit IV (COX IV expressions were observed, even though some mRNA and protein results varied. In contrast to SB216763, LiCl markedly elevated Mfn2 and COX IV protein expression levels when given together with insulin

Role of androgen receptor on cyclic mechanical stretch-regulated proliferation of C2C12 myoblasts and its upstream signals: IGF-1-mediated PI3K/Akt and MAPKs pathways.

Science.gov (United States)

Ma, Yiming; Fu, Shaoting; Lu, Lin; Wang, Xiaohui

2017-07-15

To detect the effects of androgen receptor (AR) on cyclic mechanical stretch-modulated proliferation of C2C12 myoblasts and its pathways: roles of IGF-1, PI3K and MAPK. C2C12 were randomly divided into five groups: un-stretched control, six or 8 h of fifteen percent stretch, and six or 8 h of twenty percent stretch. Cyclic mechanical stretch of C2C12 were completed using a computer-controlled FlexCell Strain Unit. Cell proliferation and IGF-1 concentration in medium were detected by CCK8 and ELISA, respectively. Expressions of AR and IGF-1R, and expressions and activities of PI3K, p38 and ERK1/2 in stretched C2C12 cells were determined by Western blot. ①The proliferation of C2C12 cells, IGF-1 concentration in medium, expressions of AR and IGF-1R, and activities of PI3K, p38 and ERK1/2 were increased by 6 h of fifteen percent stretch, while decreased by twenty percent stretch for six or 8 h ②The fifteen percent stretch-increased proliferation of C2C12 cells was reversed by AR inhibitor, Flutamide. ③The increases of AR expression, activities of PI3K, p38 and ERK1/2 resulted from fifteen percent stretch were attenuated by IGF-1 neutralizing antibody, while twenty percent stretch-induced decreases of the above indicators were enhanced by recombinant IGF-1. ④Specific inhibitors of p38, ERK1/2 and PI3K all decreased the expression of AR in fifteen percent and twenty percent of stretched C2C12 cells. Cyclic mechanical stretch modulated the proliferation of C2C12 cells, which may be attributed to the alterations of AR via IGF-1-PI3K/Akt and IGF-1-MAPK (p38, ERK1/2) pathways in C2C12 cells. Copyright © 2017 Elsevier B.V. All rights reserved.

miR-25-3p, Positively Regulated by Transcription Factor AP-2α, Regulates the Metabolism of C2C12 Cells by Targeting Akt1

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Feng Zhang

2018-03-01

Full Text Available miR-25, a member of the miR-106b-25 cluster, has been reported as playing an important role in many biological processes by numerous studies, while the role of miR-25 in metabolism and its transcriptional regulation mechanism remain unclear. In this study, gain-of-function and loss-of-function assays demonstrated that miR-25-3p positively regulated the metabolism of C2C12 cells by attenuating phosphoinositide 3-kinase (PI3K gene expression and triglyceride (TG content, and enhancing the content of adenosine triphosphate (ATP and reactive oxygen species (ROS. Furthermore, the results from bioinformatics analysis, dual luciferase assay, site-directed mutagenesis, qRT-PCR, and Western blotting demonstrated that miR-25-3p directly targeted the AKT serine/threonine kinase 1 (Akt1 3′ untranslated region (3′UTR. The core promoter of miR-25-3p was identified, and the transcription factor activator protein-2α (AP-2α significantly increased the expression of mature miR-25-3p by binding to its core promoter in vivo, as indicated by the chromatin immunoprecipitation (ChIP assay, and AP-2α binding also downregulated the expression of Akt1. Taken together, our findings suggest that miR-25-3p, positively regulated by the transcription factor AP-2α, enhances C2C12 cell metabolism by targeting the Akt1 gene.

Particle-gamma coincidence measurements in 12C+12C and 12C+Pb collisions at 2.1 GeV/nucleon incident energy

International Nuclear Information System (INIS)

Roche, G.; Koontz, R.; Mulera, T.; Pugh, H.G.; Schroeder, L.S.; Hallman, T.; Madansky, L.; Carroll, J.; Chang, C.C.; Kirk, P.N.; Krebs, G.; Vicente, J.

1985-01-01

A particle-gamma coincidence experiment has been performed with a 2.1 GeV per nucleon 12 C beam from the Bevalac. Data were taken with C and Pb targets. The γ-ray spectra are almost independent of the energy or the kind of charged particles detected in coincidence, mainly protons and deuterons. These γ-ray spectra are interpreted as resulting from π 0 decay, and are consistent with known π 0 production rates. A search for a possible decay of singly-charged anomalons into a gamma and a deuteron (or unbound proton-neutron system) has been done by studying the γp and γd invariant mass distributions. The upper limits for such a process are found to be 2 to 20% of the deuteron production rate, for anomalon masses for 200 to 400 MeV above the deuteron mass, with an anomalon mean lifetime of up to 10 -9 s, depending on which kind of decay process is considered. (orig.)

Preparation of no-carrier-added [1-11C]ethylene and [1-11C]1,2-dibromoethane as new labelling agents

International Nuclear Information System (INIS)

Shah, F.; Pike, V.W.; Dowsett, K.

1997-01-01

A method is described for the preparation of NCA [1- 11 C] ethylene based on the passage of [1- 11 C]ethanol over heated (550 o C) quartz glass in a stainless steel tube (in preference to dehydration by catalysis on γ-alumina or pyrolysis). The [1- 11 C]ethanol is prepared from cyclotron-produced NCA [ 11 C]carbon dioxide by 11 C-carboxylation of methylmagnesium bromide, freshly prepared in dibutyl ether, and reduction of the adduct with lithium aluminium hydride in diglyme. The use of involatile solvents avoids the formation of carrier ethylene and radioactive and stable diethyl ether by cracking processes over the heated catalyst. The preparation takes 21 min from the end of radionuclide production and has a radiochemical yield of 44%, decay-corrected from [ 11 C]carbon dioxide. NCA [1- 11 C] ethylene is converted quantitatively into [1- 11 C]1,2-dibromoethane when collected in a solution of bromine in carbon tetrachloride. The NCA [1- 11 C]ethylene and [1- 11 C]1,2-dibromoethane may serve as new and useful labelling agents. (Author)

Up-Regulation of CYP2C19 Expression by BuChang NaoXinTong via PXR Activation in HepG2 Cells.

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Hong Sun

Full Text Available Cytochrome P450 2C19 (CYP2C19 is an important drug-metabolizing enzyme (DME, which is responsible for the biotransformation of several kinds of drugs such as proton pump inhibitors, platelet aggregation inhibitors and antidepressants. Previous studies showed that Buchang NaoXinTong capsules (NXT increased the CYP2C19 metabolic activity in vitro and enhanced the antiplatelet effect of clopidogrel in vivo. However, the underlying molecular mechanism remained unclear. In the present study, we examined whether Pregnane X receptor (PXR plays a role in NXT-mediated regulation of CYP2C19 expression.We applied luciferase assays, real-time quantitative PCR (qPCR, Western blotting and cell-based analysis of metabolic activity experiments to investigate the NXT regulatory effects on the CYP2C19 promoter activity, the mRNA/ protein expression and the metabolic activity.Our results demonstrated that NXT significantly increased the CYP2C19 promoter activity when co-transfected with PXR in HepG2 cells. Mutations in PXR responsive element abolished the NXT inductive effects on the CYP2C19 promoter transcription. Additionally, NXT incubation (150 and 250μg/mL also markedly up-regulated endogenous CYP2C19 mRNA and protein levels in PXR-transfected HepG2 cells. Correspondingly, NXT leaded to a significant enhancement of the CYP2C19 catalytic activity in PXR-transfected HepG2 cells.In summary, this is the first study to suggest that NXT could induce CYP2C19 expression via PXR activation.

Suppression of X-ray induced transformation by vitamin E in mouse C3H/10T1/2 cells

International Nuclear Information System (INIS)

Radner, B.S.; Kennedy, A.R.

1986-01-01

Vitamin E (d-α-tocopherol) was shown to decrease X-ray induced transformation in mouse C3H/10 1/2 cells. The d-α-tocopherol was active in the form of succinate diluted in ethanol, but was inactive at the highest non-toxic concentration of the pure substance dissolved in oil and diluted in acetone. Vitamin E succinate was effective when present only for the early portion of the radiation transformation assay period, indicating that its effect may be reversible. Vitamin E did not supress the growth and expression of transformed C3H/10 1/2 cells as foci when transformed cells were surrounded by a monolayer of normal cells. (author)

Hes6 is required for actin cytoskeletal organization in differentiating C2C12 myoblasts

Energy Technology Data Exchange (ETDEWEB)

Malone, Caroline M.P.; Domaschenz, Renae; Amagase, Yoko [MRC Cancer Cell Unit, Hutchison-MRC Research Centre, Addenbrooke' s Hospital, Cambridge CB2 0XZ (United Kingdom); Dunham, Ian [EMBL-European Bioinformatics Institute (EBI), Wellcome Trust Genome Campus, Hinxton, Cambridge CB10 1SD (United Kingdom); Murai, Kasumi [MRC Cancer Cell Unit, Hutchison-MRC Research Centre, Addenbrooke' s Hospital, Cambridge CB2 0XZ (United Kingdom); Jones, Philip H., E-mail: phj20@cam.ac.uk [MRC Cancer Cell Unit, Hutchison-MRC Research Centre, Addenbrooke' s Hospital, Cambridge CB2 0XZ (United Kingdom)

2011-07-01

Hes6 is a member of the hairy-enhancer-of-split family of transcription factors that regulate proliferating cell fate in development and is known to be expressed in developing muscle. Here we investigate its function in myogenesis in vitro. We show that Hes6 is a direct transcriptional target of the myogenic transcription factors MyoD and Myf5, indicating that it is integral to the myogenic transcriptional program. The localization of Hes6 protein changes during differentiation, becoming predominantly nuclear. Knockdown of Hes6 mRNA levels by siRNA has no effect on cell cycle exit or induction of myosin heavy chain expression in differentiating C2C12 myoblasts, but F-actin filament formation is disrupted and both cell motility and myoblast fusion are reduced. The knockdown phenotype is rescued by expression of Hes6 cDNA resistant to siRNA. These results define a novel role for Hes6 in actin cytoskeletal dynamics in post mitotic myoblasts.

Hes6 is required for actin cytoskeletal organization in differentiating C2C12 myoblasts

International Nuclear Information System (INIS)

Malone, Caroline M.P.; Domaschenz, Renae; Amagase, Yoko; Dunham, Ian; Murai, Kasumi; Jones, Philip H.

2011-01-01

Hes6 is a member of the hairy-enhancer-of-split family of transcription factors that regulate proliferating cell fate in development and is known to be expressed in developing muscle. Here we investigate its function in myogenesis in vitro. We show that Hes6 is a direct transcriptional target of the myogenic transcription factors MyoD and Myf5, indicating that it is integral to the myogenic transcriptional program. The localization of Hes6 protein changes during differentiation, becoming predominantly nuclear. Knockdown of Hes6 mRNA levels by siRNA has no effect on cell cycle exit or induction of myosin heavy chain expression in differentiating C2C12 myoblasts, but F-actin filament formation is disrupted and both cell motility and myoblast fusion are reduced. The knockdown phenotype is rescued by expression of Hes6 cDNA resistant to siRNA. These results define a novel role for Hes6 in actin cytoskeletal dynamics in post mitotic myoblasts.

Bonding wood-saxon potential and the mechanism of resonance states in the ''1''2C+''1''2C system

International Nuclear Information System (INIS)

Kim, G.; Khaydarov, R.R.

2001-01-01

In present work the ''1''2C+''1''2C system are investigated in the realistic Woods--Saxon potential with Coulomb interaction. The comparison of the calculated states with the experimental data has shown, that the observed (identified) resonances may be explained by the single-channel description, i.e., as potential resonances. The quadrupole moments and transition probabilities for low-laying states have been calculated

Suppression of transformed foci, induced by alpha radiation of C3H 10T1/2 cells, by untransformed cells

International Nuclear Information System (INIS)

Lloyd, E.L.; Gemmell, M.A.; Henning, C.B.

1978-01-01

The C3H 10T1/2 CL8 cell line obtained from a mouse embryo has been widely used for screening chemical carcinogens. Transformed foci are easily distinguishable in this system as crisscrossed, piled-up cells which stain more deeply than the surrounding untransformed cells. When these foci are ringcloned and subcultured, they have been shown to give rise to malignant tumors in C3H immunodepressed mice. Previous work showed that such malignant transformations, which occurred with a dose dependent frequency, could be induced by alpha particle irradiation. The present study, in turn, demonstrates that the expression of these transformations can be completely suppressed by co-cultivating the transformed cells with a large number of untransformed cells. The precise ratio of the number of untransformed cells to transformed cells to give complete suppression was found to vary in different experiments. Maximum effects were seen when a small number of transformed cells in low passage were used. These experiments may provide at least a partial explanation for the greatly increased frequency of transformations per cell irradiated in vitro, compared with the number of tumors observed after irradiation of the same number of cells in vivo. In addition, if conditions could be optimized whereby transformed foci could reproducibly be eliminated by the use of a known number of untransformed cells, this might have important applications in the prevention and treatment of certain human cancers

Quasi-exclusive measurement of /sup 12/C(/sup 12/C, 3. cap alpha. )X at 2. 1 GeV/nucleon

Energy Technology Data Exchange (ETDEWEB)

Engelage, J; Baumgartner, M; Greiner, D E; Lindstrom, P J; Olson, D L; Wada, R; Crawford, H J; Webb, M L

1986-05-29

A study of the reaction /sup 12/C(/sup 12/C,3..cap alpha..)X at 2.1 GeV/nucleon has been completed. The energy and momentum transferred to the /sup 12/C projectile and the cross section for the dissociation of /sup 12/C into three alpha particles have been measured, 9.7 (+5.0/-2.5) millibarns. It is found that the results from this analysis are inconsistent with the predictions of current theoretical models for peripheral relativistic heavy ion collisions.

On centrality and rapidity dependences of transverse momentum spectra of negative pions in 12C + 12C collisions at 4.2 GeV/c per nucleon

International Nuclear Information System (INIS)

Iqbal, A.; Haseeb, M.Q.; Olimov, K.K.; Khan, Imran; Yuldashev, B.S.

2014-01-01

The dependences of the experimental transverse momentum spectra of the negative pions, produced in minimum bias 12 C + 12 C collisions at a momentum of 4.2A GeV/c, on the collision centrality and the pion rapidity range were studied. To analyze quantitatively the change in the p t spectra of π - mesons with the changes of collision centrality and pion rapidity range, the extracted p t spectra were fitted by Hagedorn, Boltzmann, simple exponential and Gaussian functions. The values of the extracted spectral temperatures T 1 and T 2 were consistently larger for the p t spectra of π - mesons coming from midrapidity range as compared to those of the negative pions generated in the target and projectile fragmentation regions. The spectral temperatures T 1 and T 2 extracted from fitting the p t spectra of π - mesons in range p t = 0.1–1.2 GeV/c practically coincided with each other in peripheral, semicentral and central 12 C + 12 C collision events, and thus did not show any collision centrality dependence. However, the values of T 1 and T 2 extracted from fitting in range p t = 0.1–0.7 GeV/c were consistently and noticeably larger in case of central collisions as compared to peripheral and semicentral 12 C + 12 C collisions. Hagedorn and Boltzmann functions provided significantly better fits of the transverse momentum spectra of the negative pions with the physically acceptable values of the extracted temperatures as compared to Gaussian and simple exponential functions. (author)

Orosomucoid binds insulin and IGF1 and reduces hormone stimulated protein synthesis and glucose metabolism in C2C12 myotubes

Science.gov (United States)

Previous research has indicated that orosomuciod (ORM1) may enhance insulin response in 3T3-L1 adipocytes. The present study was undertaken to determine if ORM1 can modify muscle metabolism by examining glucose oxidation and protein synthesis in the C2C12 muscle cell line. Cells were used for expe...

THE GALACTIC R CORONAE BOREALIS STARS: THE C2 SWAN BANDS, THE CARBON PROBLEM, AND THE 12C/13C RATIO

International Nuclear Information System (INIS)

Hema, B. P.; Pandey, Gajendra; Lambert, David L.

2012-01-01

Observed spectra of R Coronae Borealis (RCB) and hydrogen-deficient carbon (HdC) stars are analyzed by synthesizing the C 2 Swan bands (1, 0), (0, 0), and (0, 1) using our detailed line list and the Uppsala model atmospheres. The (0, 1) and (0, 0) C 2 bands are used to derive the 12 C abundance, and the (1, 0) 12 C 13 C band to determine the 12 C/ 13 C ratios. The carbon abundance derived from the C 2 Swan bands is about the same for the adopted models constructed with different carbon abundances over the range 8.5 (C/He = 0.1%) to 10.5 (C/He = 10%). Carbon abundances derived from C I lines are about a factor of four lower than the carbon abundance of the adopted model atmosphere over the same C/He interval, as reported by Asplund et al., who dubbed the mismatch between adopted and derived C abundance as the 'carbon problem'. In principle, the carbon abundances obtained from C 2 Swan bands and that assumed for the model atmosphere can be equated for a particular choice of C/He that varies from star to star. Then, the carbon problem for C 2 bands is eliminated. However, such C/He ratios are in general less than those of the extreme helium stars, the seemingly natural relatives to the RCB and HdC stars. A more likely solution to the C 2 carbon problem may lie in a modification of the model atmosphere's temperature structure. The derived carbon abundances and the 12 C/ 13 C ratios are discussed in light of the double degenerate and the final flash scenarios.

BPN, a marine-derived PTP1B inhibitor, activates insulin signaling and improves insulin resistance in C2C12 myotubes.

Science.gov (United States)

Xu, Qi; Luo, Jiao; Wu, Ning; Zhang, Renshuai; Shi, Dayong

2018-01-01

Insulin resistance is a key feature of type 2 diabetes mellitus (T2DM) and is characterized by defects in insulin signaling. Protein tyrosine phosphatase 1B (PTP1B) is a major negative regulator of insulin signaling cascade and has attracted intensive investigation in recent T2DM therapy study. BPN, a marine-derived bromophenol compound, was isolated from the red alga Rhodomela confervoides. This study investigated the effects of BPN on the insulin signaling pathway in insulin-resistant C2C12 myotubes by inhibiting PTP1B. Molecular docking study and analysis of small- molecule interaction with PTP1B all showed BPN inhibited PTP1B activity via binding to the catalytic site through hydrogen bonds. We then found that BPN permeated into C2C12 myotubes, on the one hand, activated insulin signaling in an insulin-independent manner in C2C12 cells; on the other hand, ameliorated palmitate-induced insulin resistance through augmenting insulin sensitivity. Moreover, our studies also showed that PTP1B inhibition by BPN increased glucose uptake in normal and insulin-resistant C2C12 myotubes through glucose transporter 4 (GLUT4) translocation. Taken together, BPN activates insulin signaling and alleviates insulin resistance and represents a potential candidate for further development as an antidiabetic agent. Copyright © 2017 Elsevier B.V. All rights reserved.

17 CFR 240.15c1-2 - Fraud and misrepresentation.

Science.gov (United States)

2010-04-01

... 17 Commodity and Securities Exchanges 3 2010-04-01 2010-04-01 false Fraud and misrepresentation. 240.15c1-2 Section 240.15c1-2 Commodity and Securities Exchanges SECURITIES AND EXCHANGE COMMISSION... Securities Exchange Act of 1934 Rules Relating to Over-The-Counter Markets § 240.15c1-2 Fraud and...

Complete fusion of the 12C+12O, 14N+12C and 15N+12C systems

International Nuclear Information System (INIS)

Conjeaud, M.; Gary, S.; Harar, S.; Wieleczko, J.P.

1978-01-01

Cross sections for evaporation residues following the complete fusion of the 12 C+ 12 C, 14 N+ 12 C and 15 N+ 12 C systems have been measured with a E-ΔE counter telescope in a wide range of incident energies. They are fairly well reproduced by evaporation calculations based on the statistical theory. The total fusion excitation function of the 12 C+ 12 C system shows strong structure, which is compared to the predictions of the reaction cross sections derived from coupled channel calculations and to the integrated inelastic cross sections. Critical angular momenta have been obtained from the fusion cross-section data and these values are discussed in the framework of compound nucleus and entrance channel effects. A striking difference is observed between the fusion cross sections of the 14 N+ 12 C and 15 N+ 12 C systems and shows the importance of the valence nucleons of colliding ions in the fusion process. A possible interpretation might be the influence of the yrast line of the compound nuclei. (Auth.)

Particle-gamma coincidence measurements in /sup 12/C+/sup 12/C and /sup 12/C+Pb collisions at 2. 1 GeV/nucleon incident energy

Energy Technology Data Exchange (ETDEWEB)

Roche, G.; Koontz, R.; Mulera, T.; Pugh, H.G.; Schroeder, L.S. (Lawrence Berkeley Lab., CA (USA)); Hallman, T.; Madansky, L. (Johns Hopkins Univ., Baltimore, MD (USA)); Carroll, J. (California Univ., Los Angeles (USA)); Chang, C.C. (Maryland Univ., College Park (USA)); Kirk, P.N.

1985-06-24

A particle-gamma coincidence experiment has been performed with a 2.1 GeV per nucleon /sup 12/C beam from the Bevalac. Data were taken with C and Pb targets. The ..gamma..-ray spectra are almost independent of the energy or the kind of charged particles detected in coincidence, mainly protons and deuterons. These ..gamma..-ray spectra are interpreted as resulting from ..pi../sup 0/ decay, and are consistent with known ..pi../sup 0/ production rates. A search for a possible decay of singly-charged anomalons into a gamma and a deuteron (or unbound proton-neutron system) has been done by studying the ..gamma..p and ..gamma..d invariant mass distributions. The upper limits for such a process are found to be 2 to 20% of the deuteron production rate, for anomalon masses for 200 to 400 MeV above the deuteron mass, with an anomalon mean lifetime of up to 10/sup -9/ s, depending on which kind of decay process is considered.

Integral measurement of the $^{12}$C(n,p)$^{12}$B reaction up to 10 GeV

CERN Document Server

Žugec, P; Bosnar, D; Ventura, A; Mengoni, A; Altstadt, S; Andrzejewski, J; Audouin, L; Barbagallo, M; Bécares, V; Bečvář, F; Belloni, F; Berthoumieux, E; Billowes, J; Boccone, V; Brugger, M; Calviani, M; Calviño, F; Cano-Ott, D; Carrapiço, C; Cerutti, F; Chiaveri, E; Chin, M; Cortés, G; Cortés-Giraldo, M.A; Cosentino, L; Diakaki, M; Domingo-Pardo, C; Dressler, R; Duran, I; Eleftheriadis, C; Ferrari, A; Finocchiaro, P; Fraval, K; Ganesan, S; García, A R; Giubrone, G; Gómez-Hornillos, M B; Gonçalves, I F; González-Romero, E; Griesmayer, E; Guerrero, C; Gunsing, F; Gurusamy, P; Heinitz, S; Jenkins, D G; Jericha, E; Käppeler, F; Karadimos, D; Kivel, N; Kokkoris, M; Krtička, M; Kroll, J; Langer, C; Lederer, C; Leeb, H; Leong, L S; Meo, S Lo; Losito, R; Manousos, A; Marganiec, J; Martínez, T; Massimi, C; Mastinu, P; Mastromarco, M; Mendoza, E; Milazzo, P M; Mingrone, F; Mirea, M; Mondalaers, W; Musumarra, A; Paradela, C; Pavlik, A; Perkowski, J; Plompen, A; Praena, J; Quesada, J; Rauscher, T; Reifarth, R; Riego, A; Roman, F; Rubbia, C; Sarmento, R; Saxena, A; Schillebeeckx, P; Schmidt, S; Schumann, D; Tagliente, G; Tain, J L; Tarrío, D; Tassan-Got, L; Tsinganis, A; Valenta, S; Vannini, G; Variale, V; Vaz, P; Versaci, R; Vermeulen, M J; Vlachoudis, V; Vlastou, R; Wallner, A; Ware, T; Weigand, M; Weiß, C; Wright, T

2016-01-01

The integral measurement of the $^{12}$C(n,p)$^{12}$B reaction was performed at the neutron time of flight facility n_TOF at CERN. The total number of $^{12}$B nuclei produced per neutron pulse of the n_TOF beam was determined using the activation technique in combination with a time of flight technique. The cross section is integrated over the n_TOF neutron energy spectrum from reaction threshold at 13.6 MeV to 10 GeV. Having been measured up to 1 GeV on basis of the $^{235}$U(n,f) reaction, the neutron energy spectrum above 200 MeV has been reevaluated due to the recent extension of the cross section reference for this particular reaction, which is otherwise considered a standard up to 200 MeV. The results from the dedicated GEANT4 simulations have been used to evaluate the neutron flux from 1 GeV up to 10 GeV. The experimental results related to the $^{12}$C(n,p)$^{12}$B reaction are compared with the evaluated cross sections from major libraries and with the predictions of different GEANT4 models, which m...

Metabolic profiling of heat or anoxic stress in mouse C2C12 myotubes using multinuclear magnetic resonance spectroscopy

DEFF Research Database (Denmark)

Straadt, Ida K; Young, Jette F; Petersen, Bent O

2010-01-01

to anaerobic metabolism due to inhibition of the aerobic pathway in the mitochondria. Conversely, lower levels of unlabeled ((12)C) lactate were apparent at increasing severity of stress, which indicate that lactate is released from the myotubes to the medium. In conclusion, the metabolites identified......In the present study, the metabolic effects of heat and anoxic stress in myotubes from the mouse cell line C2C12 were investigated by using a combination of (13)C, (1)H, and (31)P nuclear magnetic resonance (NMR) spectroscopy and enrichment with [(13)C]-glucose. Both the (13)C and the (1)H NMR...... spectra showed reduced levels of the amino acids alanine, glutamate, and aspartate after heat or anoxic stress. The decreases were smallest at 42 degrees C, larger at 45 degrees C, and most pronounced after anoxic conditions. In addition, in both the (1)H and the (31)P NMR spectra, decreases in the high...

Study for the charge symmetric systems, 12C+13N and 12C+13C with the orthogonalized coupled-reaction-channel method

International Nuclear Information System (INIS)

Imanishi, B.; Denisov, V.; Motobayashi, T.

1996-10-01

The charge-symmetric scattering systems, 12 C+ 13 N and 12 C+ 13 C have been investigated by using the orthogonalized coupled-reaction-channel (OCRC) method with the basis functions of the elastic, inelastic and transfer channels defined by the single-particle states, 1p1/2, 2s1/2, 1d5/2 and 1d3/2 of the valence nucleon in 13 N or 13 C. The data of the elastic scattering of 13 N on 12 C measured by Lienard et al. have been explained consistently with the data of the elastic and inelastic scattering of the 12 C+ 13 C system. The CRC effects both on the above systems are very strong, although those on the 12 C+ 13 N system are fairly weaker than the 12 C+ 13 C system. The role of the highly excited single-particle states 1d3/2 is particularly important in the formation of a specific CRC scheme, i.e., the formation of the covalent molecules due to the hybridization caused by the mixing of the different parity single-particle states. The fusion cross sections of the 12 C+ 13 C system at energies below the Coulomb barrier are strongly enhanced as a result of the strong CRC effects as compared with those of the 12 C+ 12 C system, while in 12 C+ 13 N system the enhancement of the sub-barrier fusion has not been observed. The above absorption mechanism for the 12 C+ 13 C system explains the lack of the molecular-resonance phenomena observed in the 12 C+ 12 C system. We check the effects of the dipole (E1) transition of the valence nucleon in 13 N (and also in 13 C) due to the core-core Coulomb interaction in the scattering at sub-barrier energies. The effects are not appreciable. (author)

MicroRNA-27a Is Induced by Leucine and Contributes to Leucine-Induced Proliferation Promotion in C2C12 Cells

Directory of Open Access Journals (Sweden)

Guangmang Liu

2013-07-01

Full Text Available Leucine, a branched chain amino acid, is well known to stimulate protein synthesis in skeletal muscle. However, the role of leucine in myoblast proliferation remains unclear. In this study, we found that leucine could promote proliferation of C2C12 cells. Moreover, expressions of miR-27a and myostatin (a bona fide target of miR-27a were upregulated and downregulated, respectively, following leucine treatment. We also found that miR-27a loss-of-function by transfection of a miR-27a inhibitor suppressed the promotion of myoblast proliferation caused by leucine. Our results suggest that miR-27a is induced by leucine and contributes to leucine-induced proliferation promotion of myoblast.

cAMP-dependent cell differentiation triggered by activated CRHR1 in hippocampal neuronal cells.

Science.gov (United States)

Inda, Carolina; Bonfiglio, Juan José; Dos Santos Claro, Paula A; Senin, Sergio A; Armando, Natalia G; Deussing, Jan M; Silberstein, Susana

2017-05-16

Corticotropin-releasing hormone receptor 1 (CRHR1) activates the atypical soluble adenylyl cyclase (sAC) in addition to transmembrane adenylyl cyclases (tmACs). Both cAMP sources were shown to be required for the phosphorylation of ERK1/2 triggered by activated G protein coupled receptor (GPCR) CRHR1 in neuronal and neuroendocrine contexts. Here, we show that activated CRHR1 promotes growth arrest and neurite elongation in neuronal hippocampal cells (HT22-CRHR1 cells). By characterising CRHR1 signalling mechanisms involved in the neuritogenic effect, we demonstrate that neurite outgrowth in HT22-CRHR1 cells takes place by a sAC-dependent, ERK1/2-independent signalling cascade. Both tmACs and sAC are involved in corticotropin-releasing hormone (CRH)-mediated CREB phosphorylation and c-fos induction, but only sAC-generated cAMP pools are critical for the neuritogenic effect of CRH, further highlighting the engagement of two sources of cAMP downstream of the activation of a GPCR, and reinforcing the notion that restricted cAMP microdomains may regulate independent cellular processes.

Search for 12 C+ 12 C clustering in 24 Mg ground state

Indian Academy of Sciences (India)

Home; Journals; Pramana – Journal of Physics; Volume 88; Issue 2. Search for 12C+12C clustering in 24Mg ground state. B N JOSHI ARUN K JAIN D C BISWAS B V JOHN Y K GUPTA L S DANU R P VIND G K PRAJAPATI S MUKHOPADHYAY A SAXENA. Regular Volume 88 Issue 2 February 2017 Article ID 29 ...

Excited states in 22Mg via the 12C(12C,2n)22Mg reaction

International Nuclear Information System (INIS)

Jewett, Cybele; Baktash, Cyrus; Bardayan, Daniel W.; Blackmon, Jeff C.; Chipps, K.; Galindo-Uribarri, Alfredo; Greife, U.; Gross, Carl J.; Jones, K. L.; Liang, Junjien; Livesay, Jake; Kozub, R. L.; Nesaraja, Caroline D; Radford, David C.; Sarazin, F.; Smith, Michael Scott; Thomas, J. S.; Yu, Chang-Hong

2007-01-01

The 12C(12C, 2n)22Mg reaction was measured with the CLARION array and the RMS separator at the Holifield Facility of Oak Ridge National Laboratory. This experiment was performed to gather more information on the excited states in 22Mg, which might be of relevance to recent radioactive ion beam measurements of the astrophysically important 21Na(p,γ)22Mg reaction. The results are compared to direct measurements, transfer experiments and a competing experiment performed with Gammasphere

CO2 and 12C:13C Isotopic Ratios on Phoebe and Iapetus

Science.gov (United States)

Clark, R. N.; Brown, R. H.; Cruikshank, D. P.

2016-12-01

Cassini VIMS has obtained spatially resolved 0.35 to 5.1 micron reflectance spectra of Saturn's satellites beginning with the Phoebe fly-by in 2004 and a close fly-by of Iapetus in 2007. Both surfaces contain relatively abundant CO2. The new (2016 RC19) calibration of VIMS has provided a significant increase in the data quality, such that isotopic absorption bands in CO2 are now well defined. CO2 on Saturn's icy satellites is trapped (Cruikshank et al., 2010, Icarus v206 p561; Pinilla-Alonso et al. 2011, Icarus v211, p75i), predominantly in the dark material (Clark et al. 2012, Icarus v218 p831). Clark et al. modeled the CO2 abundance as 2.8% on Iapetus and 3.7% on Phoebe. The main 12CO2 band in VIMS spectra on Iapetus occurs at 4.253 microns and Phoebe at 4.266 microns. The 13CO2 absorption is strong on Phoebe at 4.367 microns and weak on Iapetus at 4.387 microns. Converting the Phoebe, Iapetus, and a lab reflectance spectrum (of trace H2O-CO2 mixture on a diffuse substrate), we derive preliminary values for the ratio of the equivalent widths of the 12C and 13C absorptions as 19±2 on Phoebe, 82±8 on Iapetus, and 98±10 for the laboratory spectrum. These ratios are related to the 12C/13C ratio, but there may be effects due to intra-molecular and inter-molecular coupling that will contribute to systematic errors in the isotopic abundances derived using equivalent-width measurements that we've yet to quantify. We Believe the effects are small, and will be attempting to quantify them in the future. For comparison, the terrestrial value of the 12C/13C ratio is 90.17, and vibrational coupling may explain the slightly high lab mixture result. The local interstellar medium is 69±15 (Boogert et al., 2000, A&A). Because the CO2 bands on Phoebe and Iapetus dark material have different positions, and because the observed 13C absorption strengths are so different, the surface evolutions must be different. The large enrichment in 13C on Phoebe argues for significant

Spinal instrumentation for unstable C1-2 injury.

Science.gov (United States)

Mizuno, J; Nakagawa, H

1999-06-01

Seventeen patients with unstable C1-2 injuries were treated between 1990 and 1997. Various methods of instrumentation surgery were performed in 16 patients, excluding a case of atlantoaxial rotatory fixation. Posterior stabilization was carried out in 14 cases using Halifax interlaminar clamp, Sof'wire or Danek cable, or more recently, transarticular screws. Transodontoid anterior screw fixation was performed in four cases of odontoid process fractures, with posterior instrumentation in two cases because of malunion. Rigid internal fixation by instrumentation surgery for the unstable C1-2 injury avoids long-term application of a Halo brace and facilitates early rehabilitation. However, the procedure is technically demanding with the risk of neural and vascular injuries, particularly with posterior screw fixation. Sagittal reconstruction of thin-sliced computed tomography scans at the C1-2 region, neuronavigator, and intraoperative fluoroscopy are essential to allow preoperative surgical planning and intraoperative guidance.

Double-strand break induction and DNA damage response after {sup 12}C ion and photon radiation in U87 glioblastoma cells; Doppelstrangbruch-Induktion und DNA-Schadensantwort nach {sup 12}C-Ionen- und Photonenstrahlung in U87 Glioblastomzellen

Energy Technology Data Exchange (ETDEWEB)

Lopez Perez, Ramon

2015-04-22

for {sup 12}C ion compared to photon radiation. Pan-nuclear γH2AX caused by ionizing radiation can be linked to the induction of small DNA fragments. These results indicated that the clustering of DSBs, rather than their sheer number, is decicive for the high relative biological effectiveness of {sup 12}C ion radiation. Further hints for clustered DSBs after {sup 12}C ion radiation were found by application of superresolution microscopy to analyze repair foci after immunofluorescent staining of γH2AX and the HRR enzyme pBRCA1. Therefore, cells were irradiated with individual or only few {sup 12}C ions under a flat angle. With the help of structured illumination microscopy (SIM), the foci and their colocalization could be measured with high accuracy beyond the classical resolution limit in 3D. Besides primary foci induced along the ion trajectory, new secondary foci appeared after 26h. They were scattered offside the ion track and were presumably caused by unrepaired non-DSB lesions in the course of DNA replication. In accordance with the dependency of such secondary DSBs on repair by break-induced replication, a HRR subpathway, colocalization between γH2AX and pBRCA1 was particularly high in the secondary foci. The inner structure of γH2AX foci was further analyzed with SPDM{sub Phymod}, another superresolution microscopy technique. This revealed that the foci, that can reach sizes up to more than 1,5 μm{sup 3}, possess a fine substructure rather than being homogenous. The results indicated that the foci are composed of several subfoci in the range of 100 nm with partially elongated shape. In accordance with the organization of chromatin in densely packed loops with interchromatin spaces, different structures and distances between the subfoci could be observed.

Inference of past atmospheric delta13C and P/sub CO2/ from 13C/12C measurements in tree rings

International Nuclear Information System (INIS)

Leavitt, S.W.

1982-01-01

Carbon dioxide release from fossil-fuel burning is significant enough that we may soon experience perceptible changes in climate with important human consequences. An accurate reconstruction of past 13 C/ 12 C ratios of atmospheric CO 2 may provide key constraints on the historical activity of the biosphere as CO 2 source or sink. Tree rings appear to be a repository of this information but there is much noise in the collection of previous reconstructions, presumably associated with site selection, radial variability, choice of representative wood chemical constituent, and subtle effects of climate on fractionation. This study attempts to avoid these pitfalls and develop a 50-yr delta 13 C/sub ATM/ record from juniper trees (genus Juniperus), in fact, by taking advantage of the influence of climate on fractionation. Trees were harvested from suitable sites in close proximity to weather stations with monthly records of temperature and precipitation. The most useful relationships for at most 7 of the 10 sites were delta 13 C with December temperature or precipitation, because the coefficients were nearly constant from one interval to the next and the intercepts differed. Local pollution effects are believed responsible for the three anomalous sites. The separation of these regression lines of different intervals is interpreted as the response of the trees to the changing delta 13 C of atmospheric CO 2 so that delta 13 C/sub ATM/ curves are constructed from this spacing. The shape of the best-fit reconstruction suggests the biosphere has acted as CO 2 source to about 1965 and may now be a net sink

[2,4-(13)C]β-hydroxybutyrate metabolism in astrocytes and C6 glioblastoma cells.

Science.gov (United States)

Eloqayli, Haytham; Melø, Torun M; Haukvik, Anne; Sonnewald, Ursula

2011-08-01

This study was undertaken to determine if the ketogenic diet could be useful for glioblastoma patients. The hypothesis tested was whether glioblastoma cells can metabolize ketone bodies. Cerebellar astrocytes and C6 glioblastoma cells were incubated in glutamine and serum free medium containing [2,4-(13)C]β-hydroxybutyrate (BHB) with and without glucose. Furthermore, C6 cells were incubated with [1-(13)C]glucose in the presence and absence of BHB. Cell extracts were analyzed by mass spectrometry and media by (1)H magnetic resonance spectroscopy and HPLC. Using [2,4-(13)C]BHB and [1-(13)C]glucose it could be shown that C6 cells, in analogy to astrocytes, had efficient mitochondrial activity, evidenced by (13)C labeling of glutamate, glutamine and aspartate. However, in the presence of glucose, astrocytes were able to produce and release glutamine, whereas this was not accomplished by the C6 cells, suggesting lack of anaplerosis in the latter. We hypothesize that glioblastoma cells kill neurons by not supplying the necessary glutamine, and by releasing glutamate.

Effects of IGFBP-2 on proliferation and differentiation in neural stem cell line C17.2

Directory of Open Access Journals (Sweden)

Deng Y

2017-07-01

Full Text Available Yujia Deng,1 Lei Wang,1,2 Lite Ge,1,3 Da Duan,1 Yi Zhuo,1 Ting Yuan,1 Weiping Yan,1 Peiqi Huang,1 Xiaohua Teng,1 Ming Lu1,3 1Department of Neurosurgery, The Second Affiliated Hospital of Hunan Normal University (163 Hospital of the People’s Liberation Army, Changsha, 2Department of Neurosurgery, Affiliated Haikou Hospital, Xiangya School of Central South University, Haikou, 3Key Laboratory of Protein Chemistry and Developmental Biology of Ministry of Education, College of Life Sciences, Hunan Normal University, Changsha, People’s Republic of China Objective: Insulin-like growth factor binding protein-2 (IGFBP-2, a member of a highly conserved family of six insulin-like growth factor binding proteins (IGFBPs, can regulate several cellular processes through IGF-dependent or IGF-independent pathway. Recent studies have provided solid evidence for the importance to delineate that olfactory ensheathing cells (OEC-conditioned medium (OCM can not only facilitate the differentiation of neural stem cell line (C17.2 into neurons, but also promote the survival and proliferation. We have previously reported that IGFBP-2 was detected in OCM. This study is designed to investigate the roles of IGFBP-2 for the regulation of C17.2 differentiation and proliferation.Methods and results: IGFBP-2 was identified and upregulated in OCM to compare with astrocytes-conditioned medium by shotgun proteomics and semiquantitative proteomic analysis. In order to investigate whether exogenous IGFBP-2 could stimulate proliferation in C17.2 cells and differentiate it into glia or neuron, we used various concentrations of IGFBP-2 to induce C17.2 cells which were cultured in DMEM/F12. The results showed that exogenous IGFBP-2 can promote proliferation in C17.2 cells, but had little effect on differentiation. Interestingly, we also found that IGFBP-2 could induce C17.2 cells to differentiate into astrocytes, while inhibiting their differentiation into neurons in a dose

Reassessment of the C-13/C-12 and C-14/C-12 isotopic fractionation ratio and its impact on high-precision radiocarbon dating

NARCIS (Netherlands)

Fahrni, Simon M.; Southon, John R.; Santos, Guaciara M.; Palstra, Sanne W. L.; Meijer, Harro A. J.; Xu, Xiaomei

2017-01-01

The vast majority of radiocarbon measurement results (C-14/C-12 isotopic ratios or sample activities) are corrected for isotopic fractionation processes (measured as C-13/C-12 isotopic ratios) that occur in nature, in sample preparation and measurement. In 1954 Harmon Craig suggested a value of 2.0

Direct Light-up of cAMP Derivatives in Living Cells by Click Reactions

Directory of Open Access Journals (Sweden)

Yan Xu

2013-10-01

Full Text Available 8-Azidoadenosine 3′,5′-cyclic monophosphate (8-azido cAMP was directly detected in living cells, by applying Cu-free azide-alkyne cycloaddition to probe cAMP derivatives by fluorescence light-up. Fluorescence emission was generated by two non-fluorescent molecules, 8-azido cAMP as a model target and difluorinated cyclooctyne (DIFO reagent as a probe. The azide-alkyne cycloaddition reaction between 8-azido cAMP and DIFO induces fluorescence in 8-azido cAMP. The fluorescence emission serves as a way to probe 8-azido cAMP in cells.

Application of inverse kinematics to 1H+23Na→12C+12C

International Nuclear Information System (INIS)

Bennett, S.J.; Clarke, N.M.; Freer, M.; Fulton, B.R.; Hall, S.J.; Karban, O.; Murgatroyd, J.T.; Tungate, G.; Gyapong, G.J.; Jarvis, N.S.; Watson, D.L.; Rae, W.D.M.; Smith, A.E.; Lilley, J.S.; Woods, P.J.; Page, R.D.

1993-01-01

Some experimental techniques used in the measurement and analysis of inverse kinematic reactions are described and applied to a kinematically complete study of the 1 H( 23 Na, 12 C) 12 C reaction at an incident energy of 180 MeV. An excitation function is obtained for the energy range of 18.5 to 19.2 MeV in 24 Mg*; equivalent to proton energies from 7.1 MeV to 7.8 MeV for the 23 Na(p, 12 C) 12 C reaction. (orig.)

C1-2 vertebral anomalies in 22q11.2 microdeletion syndrome

Energy Technology Data Exchange (ETDEWEB)

Konen, Osnat; Armstrong, Derek; Padfield, Nancy; Blaser, Susan [Hospital for Sick Children, Diagnostic Imaging, Toronto (Canada); Clarke, Howard [Hospital for Sick Children, Plastic Surgery, Toronto (Canada); Weksberg, Rosanna [Hospital for Sick Children, Clinical and Metabolic Genetics, Toronto (Canada)

2008-07-15

Chromosome 22q11.2 microdeletion syndrome (22q11DS) is characterized by cleft palate, cardiac anomalies, characteristic facies, high prevalence of skeletal anomalies and learning disability. To evaluate the prevalence of craniovertebral junction anomalies in children with 22q11DS and compare these findings to those in nonsyndromic children with velopharyngeal insufficiency (VPI). Sequential CT scans performed for presurgical carotid assessment in 76 children (45 children positive for chromosome 22q11.2 deletion and 31 negative for the deletion) with VPI were retrospectively evaluated for assessment of C1-2 anomalies. C1-2 vertebral anomalies, specifically midline C1 defects, uptilted or upswept posterior elements of C2 and fusions of C2-3, were nearly universal in our cohort of 22q11DS patients with VPI. They were strikingly absent in the majority of non-22q11DS patients with VPI. C1-2 vertebral anomalies, particularly those listed above, are important radiographic markers for 22q11DS. (orig.)

C1-2 vertebral anomalies in 22q11.2 microdeletion syndrome

International Nuclear Information System (INIS)

Konen, Osnat; Armstrong, Derek; Padfield, Nancy; Blaser, Susan; Clarke, Howard; Weksberg, Rosanna

2008-01-01

Chromosome 22q11.2 microdeletion syndrome (22q11DS) is characterized by cleft palate, cardiac anomalies, characteristic facies, high prevalence of skeletal anomalies and learning disability. To evaluate the prevalence of craniovertebral junction anomalies in children with 22q11DS and compare these findings to those in nonsyndromic children with velopharyngeal insufficiency (VPI). Sequential CT scans performed for presurgical carotid assessment in 76 children (45 children positive for chromosome 22q11.2 deletion and 31 negative for the deletion) with VPI were retrospectively evaluated for assessment of C1-2 anomalies. C1-2 vertebral anomalies, specifically midline C1 defects, uptilted or upswept posterior elements of C2 and fusions of C2-3, were nearly universal in our cohort of 22q11DS patients with VPI. They were strikingly absent in the majority of non-22q11DS patients with VPI. C1-2 vertebral anomalies, particularly those listed above, are important radiographic markers for 22q11DS. (orig.)

Inhibition of protein kinase C induces differentiation in Neuro-2a cells

International Nuclear Information System (INIS)

Minana, M.D.; Felipo, V.; Grisolia, S.

1990-01-01

1-(5-Isoquinolinylsulfonyl)-2-methylpiperazine (H7), a potent inhibitor of protein kinase C, induced neuritogenesis in Neuro-2a cells, whereas N-(2-guanidinoethyl)-5-isoquinolinesulfonamide (HA 1004), which inhibits more efficiently cAMP- and cGMP-dependent protein kinases, did not. The effect, noticeable after 3 hr, was maximum (13-fold increase at 500 μM H7) between 1 and 3 days and was maintained over 2 months. In controls, 90% of the cells were undifferentiated, whereas after 3 hr with 500 μM H7 only 25% of the cells remained undifferentiated. DNA synthesis decreased as the number of differentiated cells increased. Differentiation is also functional since acetylcholinesterase activity increased ∼7-fold after 48 hr with 500 μM H7. Phorbol 12-myristate 13-acetate, a specific activator of protein kinase C, prevented or reversed the induction of neuritogenesis and the inhibition of DNA synthesis by H7. There is a good correlation between the level of protein kinase C and the percentage of differentiated cells. The results indicate that protein kinase C may play a key role in the control of differentiation of neural cells. Some possible clinical implications are briefly discussed

Induction of apoptosis by 3-amino-6-(3-aminopropyl)-5,6-dihydro-5,11-dioxo-11H-indeno[1,2-c]isoquinoline via modulation of MAPKs (p38 and c-Jun N-terminal kinase) and c-Myc in HL-60 human leukemia cells.

Science.gov (United States)

Park, Eun-Jung; Kiselev, Evgeny; Conda-Sheridan, Martin; Cushman, Mark; Pezzuto, John M

2012-03-23

Recently, we reported that 3-amino-6-(3-aminopropyl)-5,6-dihydro-5,11-dioxo-11H-indeno[1,2-c]isoquinoline (AM6-36), sharing structural similarity with naturally occurring isoquinolines, induced activities mediated by retinoid X receptor (RXR) response element accompanied by antiproliferative effects on breast cancer cells. To further characterize the biologic potential of AM6-36, we currently report studies conducted with HL-60 human leukemia cells. AM6-36 significantly inhibited cellular proliferation in a dose- and time-dependent manner with an IC(50) value of 86 nM. When evaluated at low test concentrations (≤0.25 μM), AM6-36 induced arrest in the G2/M phase of the cell cycle. At higher concentrations (1 and 2 μM), the response shifted to apoptosis, which was consistent with the effect of AM6-36 on other apoptotic signatures including an increase of apoptotic annexin V(+) 7-AAD(-) cells, loss of mitochondrial membrane potential, induction of poly(ADP-ribose) polymerase cleavage, and activation of several caspases. These apoptotic effects are potentially due to up-regulation of p38 MAPK and JNK phosphorylation and down-regulation of c-Myc oncogene expression. Taken together, AM6-36 might serve as an effective anticancer agent by inducing G2/M cell cycle arrest and apoptosis through the activation of MAPKs and inhibition of c-Myc.

Identification of rabbit cytochromes P450 2C1 and 2C2 as arachidonic acid epoxygenases.

Science.gov (United States)

Laethem, R M; Koop, D R

1992-12-01

Microsomes prepared from COS-1 cells transiently expressing rabbit cytochromes P450 2C1 and 2C2 catalyzed the metabolism of arachidonic acid to predominantly 11,12- and 14,15-epoxyeicosatrienoic acids (EETs) when microsomal epoxide hydrolase activity was inhibited by 0.2 mM 1,2-epoxy-3,3,3-trichloropropane. P450 2C2 catalyzed the formation of 11,12-EET and 14,15-EET at a ratio of 3.0 and also produced 19-hydroxyeicosatetraenoic acid (19-HETE). The 11,12-EET, 14,15-EET, and 19-HETE represented 48.3, 15.9, and 12.8%, respectively, of the total metabolites formed. P450 2C1 produced a similar but distinct ratio of 11,12-EET to 14,15-EET (2.0) and did not produce any detectable 19-HETE. The 11,12-EET and 14,15-EET represented 63.0 and 31.1%, respectively, of the total metabolites formed. The 8,9- and 5,6-EETs were not detected with either enzyme. The ratio of the 11,12-EET to 14,15-EET was 1.5 with P450 2CAA, a P450 arachidonic acid epoxygenase (P450 2CAA) that had an amino-terminal sequence identical to that of P450 2C2 [J. Biol. Chem. 267:5552-5559 (1992)]. P450 2C1, 2C2, and 2CAA metabolized lauric acid. The ratio of omega-1- to omega-hydroxylated laurate was 3.6, 3.4, and 2.4 for P450 2CAA, P450 2C2, and P450 2C1, respectively. Purified P450 2CAA had a slightly greater apparent molecular weight than expressed P450 2C2 on sodium dodecyl sulfate-polyacrylamide gels. The results clearly establish that rabbit P450 2C1 and 2C2 are arachidonic acid epoxygenases, and they suggest that P450 2CAA and 2C2 are very similar but may not be identical isoforms.

Differential regulation of iPLA2beta splice variants by in vitro ischemia in C2C12 myotubes

DEFF Research Database (Denmark)

Poulsen, K. A.; Kolko, M.; Lambert, I. H.

2006-01-01

In this study we investigated the activity, expression and regulation of iPLA2 during ischemia in mouse C2C12 myotubes. Here, we show that in vitro ischemia, i.e. oxygen deprivation and glucose starvation, induces an iPLA2 activity that is totally reversed by siRNA knock down of iPLA2£], indicating...... preferential activation of iPLA2£]. The activity of the native iPLA2£] tetramer has in humans been proposed to be negatively regulated by interactions with catalytic inactive splice variants of the full-length protein. These variants, characterized by the presence exon 9a, have however not been identified...... of this transcript would be a C-terminally truncated î50 kDa protein lacking the catalytic site. qPCR indicated that, while the total iPLA2£] mRNA level in C2C12 myotubes increased weakly within 1-2 hours of in vitro ischemia, the transcript containing the mouse exon 9a was rapidly down regulated. In addition...

Proton spectra from 6.3 GeV/c deuteron break-up on H, D, C, Al and Bi nuclei

International Nuclear Information System (INIS)

Azhgirey, L.S.; Ignatenko, M.A.; Ivanov, V.V.; Kuznetsov, A.S.; Mescheryakov, M.G.; Razin, S.V.; Stoletov, G.D.; Vzorov, I.K.; Zhmyrov, V.N.

1977-01-01

The proton spectra from deuteron break-up on H, D, 12 C, 27 Al and 209 Bi nuclei were measured at an angle of 103 mrad (lab. system) in the momentum interval from 2.6 to 3.6 GeV/c. The measurements were made at the JINR synchrophasotron by one-arm magnetic spectrometer on-line with a computer. The extracted 6.3 GeV/c deuteron beam was incident on targets of CH 2 , CD 2 , C, Al and Bi at the thickness from 0.8 to 2 g/cm 2 . The flux of deuteron beam was equal to 5x10 8 -5x10 9 particles/pulse with a typical pulse length of 300 ms. The repetition rate was one pulse every 10 s. Proton peaks with the maximum at about 3.1 GeV/c and 280-300 MeV/c FWHM dominate in all the measured spectra. Experimental proton spectra are compared with calculations with the Reid, the Moravcsik Gartenhaus, the Hulten and the Gauss wave functions. The results, on the whole, are reasonably reproduced in the framework of the Glauber multiple scattering model taking into account the relativistic deformation of the deuteron wave function

Dielectron production in {sup 12}C+{sup 12}C collisions at 1 GeV per nucleon; Dielektronenproduktion in {sup 12}C+{sup 12}C Kollisionen bei 1 GeV pro Nukleon

Energy Technology Data Exchange (ETDEWEB)

Pachmayer, Yvonne C.

2008-08-22

This thesis contains the experimental results on dielectron production in {sup 12}C+{sup 12}C collisions at 1 GeV/u recorded with HADES. Within this work, the analysis is demonstrated, showing that leptons are efficiently reconstructed and hadrons very well discriminated. The described pair analysis shows that the combinatorial background is successfully reduced and the amount of true electron-positron pairs is not decreased. After subtracting the combinatorial background, the efficiency-corrected and normalized invariant-mass, transverse momentum and rapidity distributions of the dileptons are investigated. In the invariant-mass region 0.2-0.6 GeV/c{sup 2} the measured pair yield shows a strong excess above the contribution expected from hadron decays after freeze-out according to predictions. Together with the results obtained in {sup 12}C+{sup 12}C at 2 GeV/u it becomes evident that the overshoot of the data increases with decreasing beam energy. A detailed analysis shows that the beam energy dependence of the excess yield, i.e. the pair yield above the known eta contribution, integrated over the 0.15-0.5 GeV/c{sup 2} mass range, scales like pion production. (orig.)

Probing the possibility of a C-12/C-13 galactic abundance gradient

International Nuclear Information System (INIS)

Hawkins, I.

1990-01-01

High S/N (equal to or greater than 500) observations of interstellar CH+ with the 3.0 m telescope were performed at Lick Observatory and with the 4.0 m telescope at CTIO, of the reddened, early-type stars HD 183143, HD 24432, and HD 157038 in an effort to probe the existence of a C-12/C-13 abundance gradient in our Galaxy. Previous very high quality optical observations of interstellar CH+ toward five stars within 1 kpc of the Sun have yielded a precise weighted mean C-12/C-13 isotope ratio of 43 plus or minus 4 (1 sigma) (Hawkins, Jura, and Meyer 1985; Hawkins and Jura 1987). The isotope ratios derived toward four lines of sight in the local ISM are uniform within 12 percent. The similarity among these carbon isotope ratios determined in diffuse clouds possessing different physical conditions precludes the possibility that the CH+ molecule is suffering from isotope selective effects in these regions. The precise C-12/C-13 derivable from high quality observations of (12)CH+ and (13)CH+ provide the unique opportunity to probe the homogeneity of the ISM in a large scale and the history of nucleosynthesis in our Galaxy. Since CH+ seems to be the most sensitive probe of C-12/C-13 in the diffuse ISM, observations toward more distant stars located up to 2.5 kpc from the Sun are the best way to study the possibility of a Galactic (C-12/C-13) abundance gradient. The researcher obtained 4232 angstrom data toward all three of the stars mentioned above, 3957 angstrom data toward HD 183143 and HD 157038, and 3745 angstrom data toward HD 157038. Because of the poorer quality of the HD 24432 spectrum, and its weaker CH+ absorption lines, the satellite (13)CH+ line was not detected, and thus only a lower limit on the C-12/C-13 ratio toward this star was obtained. The results obtained from careful reduction and analysis of the data toward these stars in the Northern and Southern Hemispheres are presented

Search for alpha particles emitted at rest in the break-up of the 12C-α-12C molecule-like configuration

International Nuclear Information System (INIS)

Scheurer, J.N.; Bertault, D.; Caussanel, M.; Quebert, J.L.; Fouan, J.P.

1978-01-01

A yield of alpha particles emitted at rest is clearly observed in 16 O+ 12 C at several incident energies. These alpha particles are detected by two methods: i) the alpha particle is considered as a missing mass in the detection of two 12 C nuclei in coincidence; ii) the alpha particle is detected at zero degree with a velocity due to centre of mass motion. Such a yield is assigned to a linear chain formation of the type 12 C-α- 12 C and an excitation function between 40 and 65 MeV is given. The emission due to Coulomb effects is emphasized in the discussion to give the chief explanation of the coincidence results

Study of the p+{sup 12}C reaction at energies up to 30 MeV

Energy Technology Data Exchange (ETDEWEB)

Harada, Masahide; Yamamoto, A.; Yoshioka, S. [Kyushu Univ., Fukuoka (Japan)] [and others

1998-03-01

Double differential cross sections of charged-particles emitted in the p+{sup 12}C reaction were measured in the energy region from 14 to 26 MeV. The observed continuous components of emitted protons and {alpha}-particles were analyzed by assuming sequential decay of intermediate reaction products and/or simultaneous breakup process. It was found that the three body simultaneous decay, p+{alpha}+{sup 8}Be, and the sequential decay via p+{sup 12}C{sup *}{sub 3-} and {alpha}+{sup 9}B{sub g.s.} are most important in the proton-induced breakup of {sup 12}C for energies up to 30 MeV. (author)

MM1+2C sporadic Creutzfeldt-Jakob disease presenting as rapidly progressive nonfluent aphasia.

Science.gov (United States)

Allegri, Ricardo F; Bartoloni, Leonardo; Iturry, Mónica; Romero, Carlos; Begué, Christián; Sevlever, Gustavo; Taratuto, Ana Lía

2014-01-01

We report a 77-year-old man, presenting with progressive aphasia as an initial symptom, who developed severe dementia over the course of 20 months. Frontal cortex PrPSc western blot was type 2 and codon 129 was MM; brain neuropathology showed cortical vacuoles with perivacuolar PrP immunostaining characteristic of MM2C. Cerebellum showed focal coarse, patchy staining in different sections of the molecular layer, diffuse fine punctuate and coarse PrP immunopositive deposits in the granule cell layer, and focal synaptic immunostaining in the molecular layer, suggestive of MM1+2C by histotyping. This clinical presentation has not yet been described in an MM1+2C subtype by histotyping.

Metabolic and morphological alterations induced by proteolysis-inducing factor from Walker tumour-bearing rats in C2C12 myotubes

International Nuclear Information System (INIS)

Yano, Claudia L; Ventrucci, Gislaine; Field, William N; Tisdale, Michael J; Gomes-Marcondes, Maria Cristina C

2008-01-01

Patients with advanced cancer suffer from cachexia, which is characterised by a marked weight loss, and is invariably associated with the presence of tumoral and humoral factors which are mainly responsible for the depletion of fat stores and muscular tissue. In this work, we used cytotoxicity and enzymatic assays and morphological analysis to examine the effects of a proteolysis-inducing factor (PIF)-like molecule purified from ascitic fluid of Walker tumour-bearing rats (WF), which has been suggested to be responsible for muscle atrophy, on cultured C 2 C 12 muscle cells. WF decreased the viability of C 2 C 12 myotubes, especially at concentrations of 20–25 μg.mL -1 . There was an increase in the content of the pro-oxidant malondialdehyde, and a decrease in antioxidant enzyme activity. Myotubes protein synthesis decreased and protein degradation increased together with an enhanced in the chymotrypsin-like enzyme activity, a measure of functional proteasome activity, after treatment with WF. Morphological alterations such as cell retraction and the presence of numerous cells in suspension were observed, particularly at high WF concentrations. These results indicate that WF has similar effects to those of proteolysis-inducing factor, but is less potent than the latter. Further studies are required to determine the precise role of WF in this experimental model

Metabolic and morphological alterations induced by proteolysis-inducing factor from Walker tumour-bearing rats in C2C12 myotubes

Directory of Open Access Journals (Sweden)

Tisdale Michael J

2008-01-01

Full Text Available Abstract Background Patients with advanced cancer suffer from cachexia, which is characterised by a marked weight loss, and is invariably associated with the presence of tumoral and humoral factors which are mainly responsible for the depletion of fat stores and muscular tissue. Methods In this work, we used cytotoxicity and enzymatic assays and morphological analysis to examine the effects of a proteolysis-inducing factor (PIF-like molecule purified from ascitic fluid of Walker tumour-bearing rats (WF, which has been suggested to be responsible for muscle atrophy, on cultured C2C12 muscle cells. Results WF decreased the viability of C2C12 myotubes, especially at concentrations of 20–25 μg.mL-1. There was an increase in the content of the pro-oxidant malondialdehyde, and a decrease in antioxidant enzyme activity. Myotubes protein synthesis decreased and protein degradation increased together with an enhanced in the chymotrypsin-like enzyme activity, a measure of functional proteasome activity, after treatment with WF. Morphological alterations such as cell retraction and the presence of numerous cells in suspension were observed, particularly at high WF concentrations. Conclusion These results indicate that WF has similar effects to those of proteolysis-inducing factor, but is less potent than the latter. Further studies are required to determine the precise role of WF in this experimental model.

Study of the alpha+12C →16O capture via 12C(16O,12C)16O transfer reaction

International Nuclear Information System (INIS)

Morais, M.C.; Lichtenthaeler, R.; Arazi, A.; Hojman, D.; Cardona, M.A.; Fimiani, L.; Carnelli, P.F.; Marti, G.V.; Pacheco, A.J.; Heimann, D. Martinez; Negri, A.E.; Capurro, O.A.; Niello, J.O. Fernandez; Montero, P.

2009-01-01

Full text: The 12 C(α γ) 16 O reaction is one of the most important in nuclear astrophysics. The rate of this reaction influences the subsequent nucleosynthesis of heavier elements in massive stars and consequently determines the stellar evolution. The 12 C(α γ) 16 O reaction cross section is extremely small at stellar temperature, this is mainly due to the Gamow energy (∼ 300KeV at T= 2x10 8 K) be much lower than the Coulomb barrier, E C.B. : = 3:142MeV. The α+ 12 C → 16 O at low energies is dominated by subthreshold states (1 - , 7:12MeV) and (2 + , 6:92MeV) of the 16 O, thus spectroscopic factors measurements of these states are very important. We propose to use the α-transfer reaction 12 C( 16 O, 12 C) 16 O * at very forward angles to obtain those spectroscopic factors. In this way, measurements of elastic and inelastic angular distributions for the 12 C+ 16 O system were performed at the TANDAR Laboratory in Buenos Aires, using an 16 O beam of E lab = 46.3MeV. These angular distributions were obtained using 8 adjacent surface barrier detectors and the measurements were performed from 5 deg to 48.5 deg with steps of 1 deg. The scattering chamber has an angular precision better than 0.1 deg. The transfer angular distributions are being analysed by DWBA using the computer code FRESCO in order to obtain the two spectroscopic factors. (author)

Raman spectroscopy of isotopically pure ({sup 12}C, {sup 13}C) and isotopically mixed ({sup 12.5}C) diamond single crystals at ultrahigh pressures

Energy Technology Data Exchange (ETDEWEB)

Enkovich, P. V., E-mail: enkovich@hppi.troitsk.ru; Brazhkin, V. V.; Lyapin, S. G.; Novikov, A. P. [Russian Academy of Sciences, Troitsk, Institute for High-Pressure Physics (Russian Federation); Kanda, H. [National Institute for Materials Science (Japan); Stishov, S. M. [Russian Academy of Sciences, Troitsk, Institute for High-Pressure Physics (Russian Federation)

2016-09-15

The Raman scattering by isotopically pure {sup 12}C and {sup 13}C diamond single crystals and by isotopically mixed {sup 12.5}C diamond single crystals is studied at a high accuracy. The studies are performed over a wide pressure range up to 73 GPa using helium as a hydrostatic pressure-transferring medium. It is found that the quantum effects, which determine the difference between the ratio of the Raman scattering frequencies in the {sup 12}C and {sup 13}C diamonds and the classical ratio (1.0408), increase to 30 GPa and then decrease. Thus, inversion in the sign of the quantum contribution to the physical properties of diamond during compression is detected. Our data suggest that the maximum possible difference between the bulk moduli of the {sup 12}C and {sup 13}C diamonds is 0.15%. The investigation of the isotopically mixed {sup 12.5}C diamond shows that the effective mass, which determines the Raman frequency, decreases during compression from 12.38 au at normal pressure to 12.33 au at 73 GPa.

IMPROVED LINE DATA FOR THE SWAN SYSTEM 12C13C ISOTOPOLOGUE

International Nuclear Information System (INIS)

Ram, Ram S.; Brooke, James S. A.; Bernath, Peter F.; Sneden, Christopher; Lucatello, Sara

2014-01-01

We present new, accurate predictions for rotational line positions, excitation energies, and transition probabilities of the 12 C 13 C isotopologue Swan d 3 Π-a 3 Π system 0-0, 0–1, 0–2, 1–0, 1–1, 1–2, 2–0, 2–1, and 2–2 vibrational bands. The line positions and energy levels were predicted through new analyses of published laboratory data for the 12 C 13 C lines. Transition probabilities were derived from recent computations of transition dipole moments and related quantities. The 12 C 13 C line data were combined with similar data for 12 C 2, reported in a companion paper, and applied to produce synthetic spectra of carbon-rich metal-poor stars that have strong C 2 Swan bands. The matches between synthesized and observed spectra were used to estimate band head positions for a few of the 12 C 13 C vibrational bands and to verify that the new computed line data match observed spectra. The much weaker C 2 lines of the bright red giant Arcturus were also synthesized in the band head regions

Absolute cross sections measurement for the 12C + 12C system at astrophysically relevant energies

International Nuclear Information System (INIS)

Barron-Palos, L.; Aguilera, E.F.; Aspiazu, J.; Huerta, A.; Martinez-Quiroz, E.; Monroy, R.; Moreno, E.; Murillo, G.; Ortiz, M.E.; Policroniades, R.; Varela, A.; Chavez, E.

2006-01-01

The 12 C + 12 C fusion reaction has been studied in the center-of-mass energy range of 2.25 to 6.01 MeV. Through the detection of gamma rays from the first excited states of the residual nuclei 20 Ne, 23 Na and 23 Mg, absolute cross sections for the 12 C( 12 C,-bar α), 12 C( 12 C,-bar p) and 12 C( 12 C,-bar n) reactions have been obtained. In this new measurement, the energy dependence of the S-factor is found to increase as the energy decreases below 3 MeV in the center of mass. This tendency was observed in previous measurements by Mazarakis et al., and has since then become a subject of controversy. In this work, where the cross sections are measured at even lower energies, we confirm the rise in the S-factor toward the energy region relevant for star evolution and nucleosynthesis calculations (E c.m. =1-3 MeV)

Structure in the 12C+ 12C and 8Be+16O fission width distribution of 24Mg observed via the reaction 12C(16O,α)24Mg(→X+Y)

International Nuclear Information System (INIS)

Lazzarini, A.J.; Steadman, S.G.; Ledoux, R.J.; Sperduto, A.; Young, G.R.; Van Bibber, K.; Cosman, E.R.

1983-01-01

Prominent gross and intermediate width structures are observed in the 12 C+ 12 C, 12 C+ 12 C((2 + ), 1 C((2 + )+ 12 C((2 + ), 8 Be+ 16 O, and 8 Be+ 16 O + (3 - , O + ) decay channels following 24 Mg* population via the 12 C( 16 O,α) 24 Mg reaction at E/sub c.m./ = 33 MeV. Evidence that the 12 C( 16 O,α) 24 Mg reaction populates states in 24 Mg which are associated with 12 C+ 12 C resonances is presented in the form of correlation analyses between the α+ 12 C+ 12 C three-body spectra and previously measured 12 C+ 12 C elastic and inelastic excitation functions. Direct determination of 12 C+ 12 C widths from these measurements is obscured by a background of other strong transitions which appear to be present in the 12 C( 16 O,α) 24 Mg singles spectrum

Radiation-induced transformation in oncogene primed C3H/10T1/2 cells; a new system for analysis of multi-step transformation in vitro

International Nuclear Information System (INIS)

Drozdoff, V.V.

1988-01-01

Several established rodent cell lines, such as C3H/10T1/2 fibroblasts, have been developed to study radiation and chemically-induced malignant transformation. Most experimental evidence has supported the idea that transformation in 10T1/2 cells involved at least two steps but that the apparent frequency of transformation depends on the density of plated cells. A new approach is presented here for studying radiation-induced transformation. An oncogene primed cell system (C3H-myc) was developed by introducing a constitutively active mouse c-myc gene into 10T1/2 cells. A primary goal was to determine if the introduction of an activated oncogene could substitute for one of the required steps in radiation-induced transformation. Results are presented that show that the expression of the exogenous myc gene significantly increased the frequency of radiation-induced transformation in these cells. Subculture experiments performed to analyze the kinetics of transformation in C3H-myc cells and reconstruction experiments allowing the effects of normal cells on radiation-induced transformants to be determined indicated that transformed cells arose very shortly after irradiation. These results support the conclusion that a radiation-induced event can complement the effect of myc in C3H-myc cells and directly result in transformation. This system thus provides an opportunity to isolate early steps in radiation-induced transformation and should facilitate the identification and analysis of these events

JAZF1 promotes proliferation of C2C12 cells, but retards their myogenic differentiation through transcriptional repression of MEF2C and MRF4—Implications for the role of Jazf1 variants in oncogenesis and type 2 diabetes

Energy Technology Data Exchange (ETDEWEB)

Yuasa, Katsutoshi; Aoki, Natsumi; Hijikata, Takao, E-mail: hijikata@musashino-u.ac.jp

2015-08-15

Single-nucleotide polymorphisms associated with type 2 diabetes (T2D) have been identified in Jazf1, which is also involved in the oncogenesis of endometrial stromal tumors. To understand how Jazf1 variants confer a risk of tumorigenesis and T2D, we explored the functional roles of JAZF1 and searched for JAZF1 target genes in myogenic C2C12 cells. Consistent with an increase of Jazf1 transcripts during myoblast proliferation and their decrease during myogenic differentiation in regenerating skeletal muscle, JAZF1 overexpression promoted cell proliferation, whereas it retarded myogenic differentiation. Examination of myogenic genes revealed that JAZF1 overexpression transcriptionally repressed MEF2C and MRF4 and their downstream genes. AMP deaminase1 (AMPD1) was identified as a candidate for JAZF1 target by gene array analysis. However, promoter assays of Ampd1 demonstrated that mutation of the putative binding site for the TR4/JAZF1 complex did not alleviate the repressive effects of JAZF1 on promoter activity. Instead, JAZF1-mediated repression of Ampd1 occurred through the MEF2-binding site and E-box within the Ampd1 proximal regulatory elements. Consistently, MEF2C and MRF4 expression enhanced Ampd1 promoter activity. AMPD1 overexpression and JAZF1 downregulation impaired AMPK phosphorylation, while JAZF1 overexpression also reduced it. Collectively, these results suggest that aberrant JAZF1 expression contributes to the oncogenesis and T2D pathogenesis. - Highlights: • JAZF1 promotes cell cycle progression and proliferation of myoblasts. • JAZF1 retards myogenic differentiation and hypertrophy of myotubes. • JAZF1 transcriptionally represses Mef2C and Mrf4 expression. • JAZF1 has an impact on the phosphorylation of AMPK.

Independent AMP and NAD signaling regulates C2C12 differentiation and metabolic adaptation.

Science.gov (United States)

Hsu, Chia George; Burkholder, Thomas J

2016-12-01

The balance of ATP production and consumption is reflected in adenosine monophosphate (AMP) and nicotinamide adenine dinucleotide (NAD) content and has been associated with phenotypic plasticity in striated muscle. Some studies have suggested that AMPK-dependent plasticity may be an indirect consequence of increased NAD synthesis and SIRT1 activity. The primary goal of this study was to assess the interaction of AMP- and NAD-dependent signaling in adaptation of C2C12 myotubes. Changes in myotube developmental and metabolic gene expression were compared following incubation with 5-aminoimidazole-4-carboxamide ribonucleotide (AICAR) and nicotinamide mononucleotide (NMN) to activate AMPK- and NAD-related signaling. AICAR showed no effect on NAD pool or nampt expression but significantly reduced histone H3 acetylation and GLUT1, cytochrome C oxidase subunit 2 (COX2), and MYH3 expression. In contrast, NMN supplementation for 24 h increased NAD pool by 45 % but did not reduce histone H3 acetylation nor promote mitochondrial gene expression. The combination of AMP and NAD signaling did not induce further metabolic adaptation, but NMN ameliorated AICAR-induced myotube reduction. We interpret these results as indication that AMP and NAD contribute to C2C12 differentiation and metabolic adaptation independently.

Search for 12 C+ 12 C clustering in 24 Mg ground state

Indian Academy of Sciences (India)

In the backdrop of many models, the heavy cluster structure of the ground state of 24 Mg has been probed experimentally for the first time using the heavy cluster knockout reaction 24 Mg( 12 C, 212 C) 12 C in thequasifree scattering kinematic domain. In the ( 12 C, 212 C) reaction, the direct 12 C-knockout cross-section was ...

Comparative Analysis of Osteogenic/Chondrogenic Differentiation Potential in Primary Limb Bud-Derived and C3H10T1/2 Cell Line-Based Mouse Micromass Cultures

Directory of Open Access Journals (Sweden)

Róza Zákány

2013-08-01

Full Text Available Murine micromass models have been extensively applied to study chondrogenesis and osteogenesis to elucidate pathways of endochondral bone formation. Here we provide a detailed comparative analysis of the differentiation potential of micromass cultures established from either BMP-2 overexpressing C3H10T1/2 cells or mouse embryonic limb bud-derived chondroprogenitor cells, using micromass cultures from untransfected C3H10T1/2 cells as controls. Although the BMP-2 overexpressing C3H10T1/2 cells failed to form chondrogenic nodules, cells of both models expressed mRNA transcripts for major cartilage-specific marker genes including Sox9, Acan, Col2a1, Snorc, and Hapln1 at similar temporal sequence, while notable lubricin expression was only detected in primary cultures. Furthermore, mRNA transcripts for markers of osteogenic differentiation including Runx2, Osterix, alkaline phosphatase, osteopontin and osteocalcin were detected in both models, along with matrix calcification. Although the adipogenic lineage-specific marker gene FABP4 was also expressed in micromass cultures, Oil Red O-positive cells along with PPARγ2 transcripts were only detected in C3H10T1/2-derived micromass cultures. Apart from lineage-specific marker genes, pluripotency factors (Nanog and Sox2 were also expressed in these models, reflecting on the presence of various mesenchymal lineages as well as undifferentiated cells. This cellular heterogeneity has to be taken into consideration for the interpretation of data obtained by using these models.

Connective tissue growth factor confers drug resistance in breast cancer through concomitant up-regulation of Bcl-xL and cIAP1.

Science.gov (United States)

Wang, Ming-Yang; Chen, Pai-Sheng; Prakash, Ekambaranellore; Hsu, Hsing-Chih; Huang, Hsin-Yi; Lin, Ming-Tsan; Chang, King-Jen; Kuo, Min-Liang

2009-04-15

Connective tissue growth factor (CTGF) expression is elevated in advanced breast cancer and promotes metastasis. Chemotherapy response is only transient in most metastatic diseases. In the present study, we examined whether CTGF expression could confer drug resistance in human breast cancer. In breast cancer patients who received neoadjuvant chemotherapy, CTGF expression was inversely associated with chemotherapy response. Overexpression of CTGF in MCF7 cells (MCF7/CTGF) enhanced clonogenic ability, cell viability, and resistance to apoptosis on exposure to doxorubicin and paclitaxel. Reducing the CTGF level in MDA-MB-231 (MDA231) cells by antisense CTGF cDNA (MDA231/AS cells) mitigated this drug resistance capacity. CTGF overexpression resulted in resistance to doxorubicin- and paclitaxel-induced apoptosis by up-regulation of Bcl-xL and cellular inhibitor of apoptosis protein 1 (cIAP1). Knockdown of Bcl-xL or cIAP1 with specific small interfering RNAs abolished the CTGF-mediated resistance to apoptosis induced by the chemotherapeutic agents in MCF7/CTGF cells. Inhibition of extracellular signal-regulated kinase (ERK)-1/2 effectively reversed the resistance to apoptosis as well as the up-regulation of Bcl-xL and cIAP1 in MCF7/CTGF cells. A neutralizing antibody against integrin alpha(v)beta(3) significantly attenuated CTGF-mediated ERK1/2 activation and up-regulation of Bcl-xL and cIAP1, indicating that the integrin alpha(v)beta(3)/ERK1/2 signaling pathway is essential for CTGF functions. The Bcl-xL level also correlated with the CTGF level in breast cancer patients. We also found that a COOH-terminal domain peptide from CTGF could exert activities similar to full-length CTGF, in activation of ERK1/2, up-regulation of Bcl-xL/cIAP1, and resistance to apoptosis. We conclude that CTGF expression could confer resistance to chemotherapeutic agents through augmenting a survival pathway through ERK1/2-dependent Bcl-xL/cIAP1 up-regulation.

Dielectron production in 12C+12C collisions at 1 GeV per nucleon

International Nuclear Information System (INIS)

Pachmayer, Yvonne C.

2008-01-01

This thesis contains the experimental results on dielectron production in 12 C+ 12 C collisions at 1 GeV/u recorded with HADES. Within this work, the analysis is demonstrated, showing that leptons are efficiently reconstructed and hadrons very well discriminated. The described pair analysis shows that the combinatorial background is successfully reduced and the amount of true electron-positron pairs is not decreased. After subtracting the combinatorial background, the efficiency-corrected and normalized invariant-mass, transverse momentum and rapidity distributions of the dileptons are investigated. In the invariant-mass region 0.2-0.6 GeV/c 2 the measured pair yield shows a strong excess above the contribution expected from hadron decays after freeze-out according to predictions. Together with the results obtained in 12 C+ 12 C at 2 GeV/u it becomes evident that the overshoot of the data increases with decreasing beam energy. A detailed analysis shows that the beam energy dependence of the excess yield, i.e. the pair yield above the known eta contribution, integrated over the 0.15-0.5 GeV/c 2 mass range, scales like pion production. (orig.)

Simulation of 200-400 MeV/u "1"2C + "1"2C elastic scattering on SHARAQ spectrometer

International Nuclear Information System (INIS)

Yu Lei; Zhang Gaolong; Terashima, S.; Le Xiaoyun; Tanihata, I.

2015-01-01

In order to further obtain the information of three-body force (TBF) from 200-400 MeV/u "1"2C + "1"2C elastic scattering, we plan to perform this experiment on a SHARAQ spectrometer. Based on the experimental condition of the Radioactive Ion Beam Factory (RIBF)-SHARAQ facility, a simulation is given to find a compromise between the better energy and angular resolutions, and higher yield by optimizing the target thickness, beam transport mode, beam intensity and angular step. From the simulation, we found that the beam quality mainly limits the improvements of energy and angular resolutions. A beam tracking system as well as a lateral and angular dispersion-matching technique are adopted to reduce the influence of beam quality. According to the two angular settings of SHARAQ as well as the expected cross sections on the basis of the theoretical model, the energy and angular resolutions, and statistical accuracy are estimated. (authors)

Changes in vegetation phenology are not reflected in atmospheric CO2 and 13 C/12 C seasonality.

Science.gov (United States)

Gonsamo, Alemu; D'Odorico, Petra; Chen, Jing M; Wu, Chaoyang; Buchmann, Nina

2017-10-01

Northern terrestrial ecosystems have shown global warming-induced advances in start, delays in end, and thus increased lengths of growing season and gross photosynthesis in recent decades. The tradeoffs between seasonal dynamics of two opposing fluxes, CO 2 uptake through photosynthesis and release through respiration, determine the influence of the terrestrial ecosystem on the atmospheric CO 2 and 13 C/ 12 C seasonality. Here, we use four CO 2 observation stations in the Northern Hemisphere, namely Alert, La Jolla, Point Barrow, and Mauna Loa Observatory, to determine how changes in vegetation productivity and phenology, respiration, and air temperature affect both the atmospheric CO 2 and 13 C/ 12 C seasonality. Since the 1960s, the only significant long-term trend of CO 2 and 13 C/ 12 C seasonality was observed at the northern most station, Alert, where the spring CO 2 drawdown dates advanced by 0.65 ± 0.55 days yr -1 , contributing to a nonsignificant increase in length of the CO 2 uptake period (0.74 ± 0.67 days yr -1 ). For Point Barrow station, vegetation phenology changes in well-watered ecosystems such as the Canadian and western Siberian wetlands contributed the most to 13 C/ 12 C seasonality while the CO 2 seasonality was primarily linked to nontree vegetation. Our results indicate significant increase in the Northern Hemisphere soil respiration. This means, increased respiration of 13 C depleted plant materials cancels out the 12 C gain from enhanced vegetation activities during the start and end of growing season. These findings suggest therefore that parallel warming-induced increases both in photosynthesis and respiration contribute to the long-term stability of CO 2 and 13 C/ 12 C seasonality under changing climate and vegetation activity. The summer photosynthesis and the soil respiration in the dormant seasons have become more vigorous which lead to increased peak-to-through CO 2 amplitude. As the relative magnitude of the increased

11 CFR 100.11 - State (2 U.S.C. 431(12)).

Science.gov (United States)

2010-01-01

... 11 Federal Elections 1 2010-01-01 2010-01-01 false State (2 U.S.C. 431(12)). 100.11 Section 100.11 Federal Elections FEDERAL ELECTION COMMISSION GENERAL SCOPE AND DEFINITIONS (2 U.S.C. 431) General Definitions § 100.11 State (2 U.S.C. 431(12)). State means each State of the United States, the District of...

The effect of eicosapentaenoic and docosahexaenoic acid on protein synthesis and breakdown in murine C2C12 myotubes

International Nuclear Information System (INIS)

Kamolrat, Torkamol; Gray, Stuart R.

2013-01-01

Highlights: ► EPA can enhance protein synthesis and retard protein breakdown in muscle cells. ► These effects were concurrent with increases in p70s6k and FOXO3a phosphorylation. ► EPA may be a useful tool in the treatment of muscle wasting conditions. -- Abstract: Eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) have been found to stimulate protein synthesis with little information regarding their effects on protein breakdown. Furthermore whether there are distinct effects of EPA and DHA remains to be established. The aim of the current study was to determine the distinct effects of EPA and DHA on protein synthesis, protein breakdown and signalling pathways in C2C12 myotubes. Fully differentiated C2C12 cells were incubated for 24 h with 0.1% ethanol (control), 50 μM EPA or 50 μM DHA prior to experimentation. After serum (4 h) and amino acid (1 h) starvation cells were stimulated with 2 mM L-leucine and protein synthesis measured using 3 H-labelled phenylalanine. Protein breakdown was measured using 3 H-labelled phenylalanine and signalling pathways (Akt, mTOR, p70S6k, 4EBP1, rps6 and FOXO3a) via Western blots. Data revealed that after incubation with EPA protein synthesis was 25% greater (P < 0.05) compared to control cells, with no effect of DHA. Protein breakdown was 22% (P < 0.05) lower, compared to control cells, after incubation with EPA, with no effect of DHA. Analysis of signalling pathways revealed that both EPA and DHA incubation increased (P < 0.05) p70s6k phosphorylation, EPA increased (P < 0.05) FOXO3a phosphorylation, with no alteration in other signalling proteins. The current study has demonstrated distinct effects of EPA and DHA on protein metabolism with EPA showing a greater ability to result in skeletal muscle protein accretion

The effect of eicosapentaenoic and docosahexaenoic acid on protein synthesis and breakdown in murine C2C12 myotubes

Energy Technology Data Exchange (ETDEWEB)

Kamolrat, Torkamol [Musculoskeletal Research Programme, Institute of Medical Sciences, University of Aberdeen, AB25 2ZD (United Kingdom); Gray, Stuart R., E-mail: s.r.gray@abdn.ac.uk [Musculoskeletal Research Programme, Institute of Medical Sciences, University of Aberdeen, AB25 2ZD (United Kingdom)

2013-03-22

Highlights: ► EPA can enhance protein synthesis and retard protein breakdown in muscle cells. ► These effects were concurrent with increases in p70s6k and FOXO3a phosphorylation. ► EPA may be a useful tool in the treatment of muscle wasting conditions. -- Abstract: Eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) have been found to stimulate protein synthesis with little information regarding their effects on protein breakdown. Furthermore whether there are distinct effects of EPA and DHA remains to be established. The aim of the current study was to determine the distinct effects of EPA and DHA on protein synthesis, protein breakdown and signalling pathways in C2C12 myotubes. Fully differentiated C2C12 cells were incubated for 24 h with 0.1% ethanol (control), 50 μM EPA or 50 μM DHA prior to experimentation. After serum (4 h) and amino acid (1 h) starvation cells were stimulated with 2 mM L-leucine and protein synthesis measured using {sup 3}H-labelled phenylalanine. Protein breakdown was measured using {sup 3}H-labelled phenylalanine and signalling pathways (Akt, mTOR, p70S6k, 4EBP1, rps6 and FOXO3a) via Western blots. Data revealed that after incubation with EPA protein synthesis was 25% greater (P < 0.05) compared to control cells, with no effect of DHA. Protein breakdown was 22% (P < 0.05) lower, compared to control cells, after incubation with EPA, with no effect of DHA. Analysis of signalling pathways revealed that both EPA and DHA incubation increased (P < 0.05) p70s6k phosphorylation, EPA increased (P < 0.05) FOXO3a phosphorylation, with no alteration in other signalling proteins. The current study has demonstrated distinct effects of EPA and DHA on protein metabolism with EPA showing a greater ability to result in skeletal muscle protein accretion.

Up-regulation of cytosolic phospholipase A2α expression by N,N-diethyldithiocarbamate in PC12 cells; involvement of reactive oxygen species and nitric oxide

International Nuclear Information System (INIS)

Akiyama, Nobuteru; Nabemoto, Maiko; Hatori, Yoshio; Nakamura, Hiroyuki; Hirabayashi, Tetsuya; Fujino, Hiromichi; Saito, Takeshi; Murayama, Toshihiko

2006-01-01

Disulfiram (an alcohol-aversive drug) and related compounds are known to provoke several side effects involving behavioral and neurological complications. N,N-diethyldithiocarbamate (DDC) is considered as one of the main toxic species of disulfiram and acts as an inhibitor of superoxide dismutase. Since arachidonic acid (AA) formation is regulated by reactive oxygen species (ROS) and related to toxicity in neuronal cells, we investigated the effects of DDC on AA release and expression of the α type of cytosolic phospholipase A 2 (cPLA 2 α) in PC12 cells. Treatment with 80-120 μM DDC that causes a moderate increase in ROS levels without cell toxicity stimulated cPLA 2 α mRNA and its protein expression. The expression was mediated by extracellular-signal-regulated kinase (ERK1/2), one of the mitogen-activated protein kinases. Treatment with N G nitro-L-arginine methyl ester (an inhibitor of nitric oxide synthase, 1 mM) and oxy-hemoglobin (a scavenger of nitric oxide, 2 mg/mL) abolished the DDC-induced responses (ERK1/2 phosphorylation and cPLA 2 α expression). We also showed DDC-induced up-regulation of the mRNA expression of lipocortin 1, an inhibitor of PLA 2 . Furthermore, DDC treatment of the cells enhanced Ca 2+ -ionophore-induced AA release in 30 min, although the effect was limited. Changes in AA metabolism in DDC-treated cells may have a potential role in mediating neurotoxic actions of disulfiram. In this study, we show the first to demonstrate the up-regulation of cPLA 2 α expression by DDC treatment in neuronal cells

Patients Lacking a KIR-Ligand of HLA Group C1 or C2 Have a Better Outcome after Umbilical Cord Blood Transplantation

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Carmen Martínez-Losada

2017-07-01

Full Text Available Donor natural killer (NK cells can destroy residual leukemic cells after allogeneic hematopoietic stem cell transplantation. This effect is based on the interaction of killer-cell immunoglobulin-like receptors (KIR of donor NK cells with ligands of the major histocompatibility complex found on the surface of the target cells. HLA-C1 subtypes provide the ligand for KIR2DL2 and KIR2DL3 and the HLA-C2 subtypes for KIR2DL1. We have studied the probability of relapse (PR after single-unit unrelated cord blood transplantation (UCBT in relation to the potential graft-vs.-leukemia effect mediated by NK cells present in the umbilical cord blood (UCB by analyzing KIR-ligand and HLA-C typing of the receptor. Data from 33 consecutive patients given a single unit UCBT were included. We have considered two groups of patients based on the absence or the presence of one of the C-ligands for inhibitory KIR and the incompatibility HLA-C1/2 between UCB and patients. Group 1 (n = 21: the patient lacks a C-ligand for inhibitory KIR present in UCB NK cells, i.e., patients homozygous C1/C1 or C2/C2. Group 2 (n = 12: patients heterozygous C1/C2 in which KIR-mediated graft-vs.-leukemia effect is not expected (presence of both C ligands for inhibitory KIR in the receptor. With a median follow-up post-UCBT of 93 months, patients with absence of a C-ligand for inhibitory KIRs (Group 1 showed a lower actuarial PR than patients with both C-ligands (group 2: 21 ± 10 vs. 68 ± 18% at 2 year and 36 ± 13 vs. 84 ± 14% at 5 years (p = 0.025, respectively. In patients with acute lymphoblastic leukemia, the 2-year PR was 36 ± 21% for group 1 and 66 ± 26% for 2 (p = 0.038. Furthermore, group 1 had a lower incidence of grades II–IV acute graft-vs.-host disease (p = 0.04. In the setting of UCBT, the absence of a C-ligand (C1 or C2 of inhibitory KIR in the patient is associated with lower PR, which is probably due to the graft

Characterization and Functional Analysis of Extracellular Vesicles and Muscle-Abundant miRNAs (miR-1, miR-133a, and miR-206 in C2C12 Myocytes and mdx Mice.

Directory of Open Access Journals (Sweden)

Yasunari Matsuzaka

Full Text Available Duchenne muscular dystrophy (DMD is a progressive neuromuscular disorder. Here, we show that the CD63 antigen, which is located on the surface of extracellular vesicles (EVs, is associated with increased levels of muscle-abundant miRNAs, namely myomiRs miR-1, miR-133a, and miR-206, in the sera of DMD patients and mdx mice. Furthermore, the release of EVs from the murine myoblast C2C12 cell line was found to be modulated by intracellular ceramide levels in a Ca2+-dependent manner. Next, to investigate the effects of EVs on cell survival, C2C12 myoblasts and myotubes were cultured with EVs from the sera of mdx mice or C2C12 cells overexpressing myomiRs in presence of cellular stresses. Both the exposure of C2C12 myoblasts and myotubes to EVs from the serum of mdx mice, and the overexpression of miR-133a in C2C12 cells in presence of cellular stress resulted in a significant decrease in cell death. Finally, to assess whether miRNAs regulate skeletal muscle regeneration in vivo, we intraperitoneally injected GW4869 (an inhibitor of exosome secretion into mdx mice for 5 and 10 days. Levels of miRNAs and creatine kinase in the serum of GW4869-treated mdx mice were significantly downregulated compared with those of controls. The tibialis anterior muscles of the GW4869-treated mdx mice showed a robust decrease in Evans blue dye uptake. Collectively, these results indicate that EVs and myomiRs might protect the skeletal muscle of mdx mice from degeneration.

The 12C+12C → 8Begs+16Ogs reaction at Ecm=27 to 36 MeV

International Nuclear Information System (INIS)

Aliotta, M.; Lattuada, M.; Spitaleri, C.

1996-01-01

The 12 C + 12 C → 8 Be gs + 16 O gs reaction has been experimentally investigated at c.m. energies between 27 and 36 MeV. A resonance is present at 32.5 MeV, i.e. at the same energy of the resonance previously observed [1] in the 12 C(0 + 2 ) + 12 C(0 + 2 ) exit channel, but its width is about three times narrower. Moreover the data indicate an enhancement of the cross section at a c.m. energy close to 29 MeV. These results are discussed and compared to the predictions of the Band Crossing Model obtained for different exit channels of the 12 C + 12 C reaction. (orig.)

Cloning of the cDNA for human 12-lipoxygenase

International Nuclear Information System (INIS)

Izumi, T.; Hoshiko, S.; Radmark, O.; Samuelsson, B.

1990-01-01

A full-length cDNA clone encoding 12-lipoxygenase was isolated from a human platelet cDNA library by using a cDNA for human reticulocyte 15-lipoxygenase as probe for the initial screening. The cDNA had an open reading frame encoding 662 amino acid residues with a calculated molecular weight of 75,590. Three independent clones revealed minor heterogeneities in their DNA sequences. Thus, in three positions of the deduced amino acid sequence, there is a choice between two different amino acids. The deduced sequence from the clone plT3 showed 65% identity with human reticulocyte 15-lipoxygenase and 42% identity with human leukocyte 5-lipoxygenase. The 12-lipoxygenase cDNA recognized a 3.0-kilobase mRNA species in platelets and human erythroleukemia cells (HEL cells). Phorbol 12-tetradecanoyl 13-acetate induced megakaryocytic differentiation of HEL cells and 12-lipoxygenase activity and increased mRNA for 12-lipoxygenase. The identity of the cloned 12-lipoxygenase was assured by expression in a mammalian cell line (COS cells). Human platelet 12-lipoxygenase has been difficult to purify to homogeneity. The cloning of this cDNA will increase the possibilities to elucidate the structure and function of this enzyme

Inhibiting c-Jun N-terminal kinase partially attenuates caffeine-dependent cell death without alleviating the caffeine-induced reduction in mitochondrial respiration in C2C12 skeletal myotubes

International Nuclear Information System (INIS)

Downs, R.M.; Hughes, M.A.; Kinsey, S.T.; Johnson, M.C.; Baumgarner, B.L.

2016-01-01

Caffeine is a widely consumed stimulant that has previously been shown to promote cytotoxic stress and even cell death in numerous mammalian cell lines. Thus far there is little information available regarding the toxicity of caffeine in skeletal muscle cells. Our preliminary data revealed that treating C2C12 myotubes with 5 mM caffeine for 6 h increased nuclear fragmentation and reduced basal and maximal oxygen consumption rate (OCR) in skeletal myotubes. The purpose of this study was to further elucidate the pathways by which caffeine increased cell death and reduced mitochondrial respiration. We specifically examined the role of c-Jun N-terminal kinase (JNK), which has previously been shown to simultaneously increase caspase-dependent cell death and reduce mitochondrial respiration in other mammalian cell lines. We found that caffeine promoted a dose-dependent increase in cell death in multinucleated myotubes but did not in mononucleated myoblasts. The addition of 10 μM Z-DEVD-FMK, a specific inhibitor of executioner caspases, completely inhibited caffeine-dependent cell death. Further, the addition of 400 μM dantrolene, a specific ryanodine receptor (RYR) inhibitor, prevented the caffeine-dependent increase in cell death and the reduction in basal and maximal OCR. We also discovered that caffeine treatment significantly increased the phosphorylation of JNK and that the addition of 30 μM SP600125 (JNKi), a specific JNK inhibitor, partially attenuated caffeine-induced cell death without preventing the caffeine-dependent reduction in basal and maximal OCR. Our results suggest that JNK partially mediates the increase in caspase-dependent cell death but does not contribute to reduced mitochondrial respiration in caffeine-treated skeletal muscle cells. We conclude that caffeine increased cell death and reduced mitochondrial respiration in a calcium-dependent manner by activating the RYR and promoting reticular calcium release. - Highlights: • Caffeine

Transcriptional regulation of miR-146b by C/EBPβ LAP2 in esophageal cancer cells

International Nuclear Information System (INIS)

Li, Junxia; Shan, Fabo; Xiong, Gang; Wang, Ju-Ming; Wang, Wen-Lin; Xu, Xueqing; Bai, Yun

2014-01-01

Highlights: • MiR-146b promotes esophageal cancer cell proliferation. • MiR-146b inhibits esophageal cancer cell apoptosis. • C/EBPβ directly binds to miR-146b promoter conserved region. • MiR-146b is up-regulated by C/EBPβ LAP2 transcriptional activation. - Abstract: Recent clinical study indicated that up-regulation of miR-146b was associated with poor overall survival of patients in esophageal squamous cell carcinoma. However, the underlying mechanism of miR-146b dysregulation remains to be explored. Here we report that miR-146b promotes cell proliferation and inhibits cell apoptosis in esophageal cancer cell lines. Mechanismly, two C/EBPβ binding motifs are located in the miR-146b promoter conserved region. Among the three isoforms of C/EBPβ, C/EBPβ LAP2 positively regulated miR-146b expression and increases miR-146b levels in a dose-dependent manner through transcription activation of miR-146b gene. Together, these results suggest a miR-146b regulatory mechanism involving C/EBPβ, which may contribute to the up-regulation of miR-146b in esophageal squamous cell carcinoma

Integral measurement of the {sup 12}C(n, p){sup 12}B reaction up to 10 GeV

Energy Technology Data Exchange (ETDEWEB)

Zugec, P.; Bosnar, D. [University of Zagreb, Department of Physics, Faculty of Science, Zagreb (Croatia); Colonna, N.; Barbagallo, M.; Mastromarco, M.; Tagliente, G.; Variale, V. [Istituto Nazionale di Fisica Nucleare, Bari (Italy); Ventura, A. [Istituto Nazionale di Fisica Nucleare, Bologna (Italy); Mengoni, A. [ENEA, Bologna (Italy); Altstadt, S.; Langer, C.; Lederer, C.; Reifarth, R.; Schmidt, S.; Weigand, M. [Johann-Wolfgang-Goethe Universitaet, Frankfurt (Germany); Andrzejewski, J.; Marganiec, J.; Perkowski, J. [Uniwersytet Lodzki, Lodz (Poland); Audouin, L.; Leong, L.S.; Tassan-Got, L. [Centre National de la Recherche Scientifique/IN2P3 - IPN, Orsay (France); Becares, V.; Cano-Ott, D.; Garcia, A.R.; Gonzalez-Romero, E.; Martinez, T.; Mendoza, E. [Centro de Investigaciones Energeticas Medioambientales y Tecnologicas (CIEMAT), Madrid (Spain); Becvar, F.; Krticka, M.; Kroll, J.; Valenta, S. [Charles University, Prague (Czech Republic); Belloni, F.; Mondalaers, W.; Plompen, A.; Schillebeeckx, P. [European Commission JRC, Institute for Reference Materials and Measurements, Geel (Belgium); Berthoumieux, E.; Fraval, K.; Gunsing, F. [CEA/Saclay - IRFU, Gif-sur-Yvette (France); Billowes, J.; Ware, T.; Wright, T. [University of Manchester, Manchester (United Kingdom); Boccone, V.; Brugger, M.; Calviani, M.; Cerutti, F.; Chiaveri, E.; Chin, M.; Ferrari, A.; Guerrero, C.; Losito, R.; Roman, F.; Rubbia, C.; Tsinganis, A.; Versaci, R.; Vlachoudis, V.; Weiss, C. [CERN, Geneva (Switzerland); Calvino, F.; Cortes, G.; Gomez-Hornillos, M.B.; Riego, A. [Universitat Politecnica de Catalunya, Barcelona (Spain); Carrapico, C.; Goncalves, I.F.; Sarmento, R.; Vaz, P. [Universidade de Lisboa, C2TN-Instituto Superior Tecnico, Lisboa (Portugal); Cortes-Giraldo, M.A.; Praena, J.; Quesada, J. [Universidad de Sevilla, Sevilla (Spain); Cosentino, L.; Finocchiaro, P. [INFN - Laboratori Nazionali del Sud, Catania (Italy); Diakaki, M.; Karadimos, D.; Kokkoris, M.; Vlastou, R. [National Technical University of Athens (NTUA), Athens (Greece); Domingo-Pardo, C.; Giubrone, G.; Tain, J.L. [CSIC-Universidad de Valencia, Instituto de Fisica Corpuscular, Valencia (Spain); Dressler, R.; Heinitz, S.; Kivel, N.; Schumann, D. [Paul Scherrer Institut, Villigen (Switzerland); Duran, I.; Tarrio, D. [Universidade de Santiago de Compostela, Santiago de Compostela (Spain); Eleftheriadis, C.; Manousos, A. [Aristotle University of Thessaloniki, Thessaloniki (Greece); Ganesan, S.; Gurusamy, P.; Saxena, A. [Bhabha Atomic Research Centre (BARC), Mumbai (India); Griesmayer, E.; Jericha, E.; Leeb, H. [Atominstitut der Oesterreichischen Universitaeten, Technische Universitaet Wien, Wien (Austria); Jenkins, D.G.; Vermeulen, M.J. [University of York, York, Heslington (United Kingdom); Kaeppeler, F. [Karlsruhe Institute of Technology (KIT), Institut fuer Kernphysik, Karlsruhe (Germany); Lo Meo, S. [Istituto Nazionale di Fisica Nucleare, Bologna (Italy); ENEA, Bologna (Italy); Massimi, C.; Mingrone, F.; Vannini, G. [Dipartimento di Fisica, Universita di Bologna (IT); INFN, Bologna (IT); Mastinu, P. [Laboratori Nazionali di Legnaro, Istituto Nazionale di Fisica Nucleare, Legnaro (IT); Milazzo, P.M. [Istituto Nazionale di Fisica Nucleare, Trieste (IT); Mirea, M. [Horia Hulubei National Institute of Physics and Nuclear Engineering - IFIN HH, Magurele (RO); Musumarra, A. [Universita di Catania, Dipartimento di Fisica e Astronomia DFA, Catania (IT); INFN-Laboratori Nazionali del Sud, Catania (IT); Paradela, C. [European Commission JRC, Institute for Reference Materials and Measurements, Geel (BE); Universidade de Santiago de Compostela, Santiago de Compostela (ES); Pavlik, A. [Faculty of Physics, University of Vienna, Wien (AT); Rauscher, T. [University of Hertfordshire, Centre for Astrophysics Research, School of Physics, Astronomy and Mathematics, Hatfield (GB); University of Basel, Department of Physics, Basel (CH); Wallner, A. [Faculty of Physics, University of Vienna, Wien (AT); Australian National University, Research School of Physics and Engineering, Canberra (AU)

2016-04-15

The integral measurement of the {sup 12}C(n, p){sup 12}B reaction was performed at the neutron time-of-flight facility nTOF at CERN. The total number of {sup 12}B nuclei produced per neutron pulse of the nTOF beam was determined using the activation technique in combination with a time-of-flight technique. The cross section is integrated over the nTOF neutron energy spectrum from reaction threshold at 13.6 MeV to 10 GeV. Having been measured up to 1GeV on basis of the {sup 235}U(n, f) reaction, the neutron energy spectrum above 200 MeV has been re-evaluated due to the recent extension of the cross section reference for this particular reaction, which is otherwise considered a standard up to 200 MeV. The results from the dedicated GEANT4 simulations have been used to evaluate the neutron flux from 1 GeV up to 10 GeV. The experimental results related to the {sup 12}C(n, p){sup 12}B reaction are compared with the evaluated cross sections from major libraries and with the predictions of different GEANT4 models, which mostly underestimate the {sup 12}B production. On the contrary, a good reproduction of the integral cross section derived from measurements is obtained with TALYS-1.6 calculations, with optimized parameters. (orig.)

Hydroxylation of methylated DNA by TET1 in chondrocyte differentiation of C3H10T1/2 cells

Directory of Open Access Journals (Sweden)

Ryo Ito

2016-03-01

Full Text Available DNA methylation is closely involved in the regulation of cellular differentiation, including chondrogenic differentiation of mesenchymal stem cells. Recent studies showed that Ten–eleven translocation (TET family proteins converted 5-methylcytosine (5mC to 5-hydroxymethylcytosine, 5-formylcytosine and 5carboxylcytosine by oxidation. These reactions constitute potential mechanisms for active demethylation of methylated DNA. However, the relationship between the DNA methylation patterns and the effects of TET family proteins in chondrocyte differentiation is still unclear. In this study, we showed that DNA hydroxylation of 5mC was increased during chondrocytic differentiation of C3H10T1/2 cells and that the expression of Tet1 was particularly enhanced. Moreover, knockdown experiments revealed that the downregulation of Tet1 expression caused decreases in chondrogenesis markers such as type 2 and type 10 collagens. Furthermore, we found that TET proteins had a site preference for hydroxylation of 5mC on the Insulin-like growth factor 1 (Igf1 promoter in chondrocytes. Taken together, we showed that the expression of Tet1 was specifically facilitated in chondrocyte differentiation and Tet1 can regulate chondrocyte marker gene expression presumably through its hydroxylation activity for DNA.

Potent and Selective Covalent Quinazoline Inhibitors of KRAS G12C

Energy Technology Data Exchange (ETDEWEB)

Zeng, Mei; Lu, Jia; Li, Lianbo; Feru, Frederic; Quan, Chunshan; Gero, Thomas W.; Ficarro, Scott B.; Xiong, Yuan; Ambrogio, Chiara; Paranal, Raymond M.; Catalano, Marco; Shao, Jay; Wong, Kwok-Kin; Marto, Jarrod A.; Fischer, Eric S.; Jänne, Pasi A.; Scott, David A.; Westover, Kenneth D.; Gray, Nathanael S. (DFCI); (UTSMC); (Harvard-Med); (NYUSM)

2017-08-01

Targeted covalent small molecules have shown promise for cancers driven by KRAS G12C. Allosteric compounds that access an inducible pocket formed by movement of a dynamic structural element in KRAS, switch II, have been reported, but these compounds require further optimization to enable their advancement into clinical development. We demonstrate that covalent quinazoline-based switch II pocket (SIIP) compounds effectively suppress GTP loading of KRAS G12C, MAPK phosphorylation, and the growth of cancer cells harboring G12C. Notably we find that adding an amide substituent to the quinazoline scaffold allows additional interactions with KRAS G12C, and remarkably increases the labeling efficiency, potency, and selectivity of KRAS G12C inhibitors. Structural studies using X-ray crystallography reveal a new conformation of SIIP and key interactions made by substituents located at the quinazoline 2-, 4-, and 7-positions. Optimized lead compounds in the quinazoline series selectively inhibit KRAS G12C-dependent signaling and cancer cell growth at sub-micromolar concentrations.

Angular correlation, spin alignment, and systematics of mis-matched {sup 12}C+{sup 12}C inelastic scattering resonances

Energy Technology Data Exchange (ETDEWEB)

Wuosmaa, A.H.; Wiedenhoever, I.; Caggiano, J.; Carpenter, M.P.; Devlin, M.; Heinz, A.; Janssens, R.V.F.; Kondev, F.; Lauritsen, T.; Sarantites, D.G.; Sobotka, L.G.; Battacharyya, P

2003-10-09

Particle gamma-ray angular correlation measurements have been used to study the spin alignment and magnetic-substate population parameters for the 2{sup +}{sub 1} (4.443 MeV) state in {sup 12}C, populated in the {sup 12}C({sup 12}C,{sup 12}C[0{sup +}{sub 2}]){sup 12}C(2{sup +}{sub 1}) inelastic scattering reaction in the vicinity of a prominent, narrow peak in the scattering excitation function. The data show a strong alignment of the spin with the orbital angular momentum, and suggest that the cross section peak corresponds to a spin 14{sup +} resonance at E{sub c.m.}=28.0 MeV. This energy is close to that where a strong peak is also observed in the 0{sup +}{sub 1}+0{sup +}{sub 2} excitation function. A comparison between the data for these two channels lends some support to recent theoretical calculations of resonance behavior for angular-momentum-mismatched channels in {sup 12}C+{sup 12}C inelastic scattering.

The role of three-body coulomb fields versus final state interactions in the decay of 12C-α-12C

International Nuclear Information System (INIS)

Quebert, J.L.; Bertault, D.; Scheurer, J.N.; Fouan, J.P.

1980-01-01

The alpha emission in 16 O + 12 C→ 12 C + α + 12 C has been thoroughly studied in the region of the rapidity plot: Ysub(α)=Ysub(c.m.). The three-body coulomb fields, as well as configurations close to alignment, account for the alpha yield which is observed. The apparent competition between direct and sequential decays is well explained by the coulomb break-up

1,2-Diacylglycerols, but not phorbol esters, activate a potential inhibitory pathway for protein kinase C in GH3 pituitary cells. Evidence for involvement of a sphingomyelinase.

Science.gov (United States)

Kolesnick, R N; Clegg, S

1988-05-15

It has been suggested that sphingoid bases may serve as physiologic inhibitors of protein kinase C. Because 1,2-diacylglycerols, but not phorbol esters, enhance sphingomyelin degradation via a sphingomyelinase in GH3 pituitary cells (Kolesnick, R. N. (1987) J. Biol. Chem. 262, 16759-16762), the effects of phorbol esters, 1,2-diacylglycerols, and sphingomyelinase on protein kinase C activation were assessed. Under basal conditions, the inactive cytosolic form of protein kinase C predominated. 1,2-Diacylglycerols stimulated transient protein kinase C redistribution to the membrane. 1,2-Dioctanoylglycerol (200 micrograms/ml) reduced cytosolic protein kinase C activity to 67% of control from 72 to 48 pmol.min-1.10(6) cells-1 and enhanced membrane-bound activity to 430% of control from 6 to 25 pmol.min-1.10(6) cells-1 after 4 min of stimulation. Thereafter, protein kinase C activity returned to the cytosol. In contrast, the phorbol ester, 12-O-tetradecanoylphorbol-13-acetate (TPA), stimulated redistribution to the membrane without return to the cytosol. Exogenous sphingomyelinase reduced membrane-bound protein kinase C activity to 30% of control, yet did not alter cytosolic activity. Sphingomyelinase, added after phorbol ester-induced redistribution was completed, restored activity to the cytosol. In these studies, TPA (10(-8) M) reduced cytosolic activity to 62% of control and elevated membrane-bound protein kinase C activity to 650% of control. Sphingomyelinase restored cytosolic activity to 84% of control and reduced membrane-bound activity to 297% of control. Similarly, the free sphingoid bases, sphingosine, sphinganine, and phytosphingosine, reversed phorbol ester-induced protein kinase C redistribution. Since 1,2-diacylglycerols activate a sphingomyelinase and sphingomyelinase action can reverse protein kinase C activation, these studies suggest that a pathway involving a sphingomyelinase might comprise a physiologic negative effector system for protein kinase C

Metabolic and morphological alterations induced by proteolysis-inducing factor from Walker tumour-bearing rats in C{sub 2}C{sub 12} myotubes

Energy Technology Data Exchange (ETDEWEB)

Yano, Claudia L; Ventrucci, Gislaine [Departamento de Fisiologia e Biofísica, Instituto de Biologia, Universidade Estadual de Campinas (UNICAMP), CP 6109, 13083-970, Campinas, São Paulo (Brazil); Field, William N; Tisdale, Michael J [Cancer Research Laboratory, Pharmaceutical Sciences Research Institute, Aston University, Birmingham, B4 7ET (United Kingdom); Gomes-Marcondes, Maria Cristina C [Departamento de Fisiologia e Biofísica, Instituto de Biologia, Universidade Estadual de Campinas (UNICAMP), CP 6109, 13083-970, Campinas, São Paulo (Brazil)

2008-01-28

Patients with advanced cancer suffer from cachexia, which is characterised by a marked weight loss, and is invariably associated with the presence of tumoral and humoral factors which are mainly responsible for the depletion of fat stores and muscular tissue. In this work, we used cytotoxicity and enzymatic assays and morphological analysis to examine the effects of a proteolysis-inducing factor (PIF)-like molecule purified from ascitic fluid of Walker tumour-bearing rats (WF), which has been suggested to be responsible for muscle atrophy, on cultured C{sub 2}C{sub 12} muscle cells. WF decreased the viability of C{sub 2}C{sub 12} myotubes, especially at concentrations of 20–25 μg.mL{sup -1}. There was an increase in the content of the pro-oxidant malondialdehyde, and a decrease in antioxidant enzyme activity. Myotubes protein synthesis decreased and protein degradation increased together with an enhanced in the chymotrypsin-like enzyme activity, a measure of functional proteasome activity, after treatment with WF. Morphological alterations such as cell retraction and the presence of numerous cells in suspension were observed, particularly at high WF concentrations. These results indicate that WF has similar effects to those of proteolysis-inducing factor, but is less potent than the latter. Further studies are required to determine the precise role of WF in this experimental model.

The CYP2E1 inhibitor DDC up-regulates MMP-1 expression in hepatic stellate cells via an ERK1/2- and Akt-dependent mechanism.

Science.gov (United States)

Liu, Tianhui; Wang, Ping; Cong, Min; Xu, Youqing; Jia, Jidong; You, Hong

2013-06-05

DDC (diethyldithiocarbamate) could block collagen synthesis in HSC (hepatic stellate cells) through the inhibition of ROS (reactive oxygen species) derived from hepatocyte CYP2E1 (cytochrome P450 2E1). However, the effect of DDC on MMP-1 (matrix metalloproteinase-1), which is the main collagen degrading matrix metalloproteinase, has not been reported. In co-culture experiments, we found that DDC significantly enhanced MMP-1 expression in human HSC (LX-2) that were cultured with hepatocyte C3A cells either expressing or not expressing CYP2E1. The levels of both proenzyme and active MMP-1 enzyme were up-regulated in LX-2 cells, accompanied by elevated enzyme activity of MMP-1 and decreased collagen I, in both LX-2 cells and the culture medium. H2O2 treatment abrogated DDC-induced MMP-1 up-regulation and collagen I decrease, while catalase treatment slightly up-regulated MMP-1 expression. These data suggested that the decrease in ROS by DDC was partially responsible for the MMP-1 up-regulation. ERK1/2 (extracellular signal-regulated kinase 1/2), Akt (protein kinase B) and p38 were significantly activated by DDC. The ERK1/2 inhibitor (U0126) and Akt inhibitor (T3830) abrogated the DDC-induced MMP-1 up-regulation. In addition, a p38 inhibitor (SB203580) improved MMP-1 up-regulation through the stimulation of ERK1/2. Our data indicate that DDC significantly up-regulates the expression of MMP-1 in LX-2 cells which results in greater MMP-1 enzyme activity and decreased collagen I. The enhancement of MMP-1 expression by DDC was associated with H2O2 inhibition and coordinated regulation by the ERK1/2 and Akt pathways. These data provide some new insights into treatment strategies for hepatic fibrosis.

Nuclear molecules in the systems 12C+12C and 16O+16O

International Nuclear Information System (INIS)

Tiereth, W.

1986-01-01

For the two heavy ion systems 12 C+ 12 C and 16 O+ 16 O by means of scattering experiments studies on the phenomena of nuclear molecules were performed. For the 12 C+ 12 C system in the range of the Coulomb wall precision measurements of the elastic scattering were performed. In the two energy ranges of 5.5 ≤ E c.m. ≤ 6.65 MeV and 6.95 ≤ E c.m ≤ 7.375 MeV where the existence of many quasimolecular resonances is known in energy steps of ΔE c.m. = 25 keV angular distributions in the range 10 0 ≤ θ c.m. ≤ 90 0 with Δθ c.m. = 1 0 were measured. For several of the found quasimolecular resonances the spin and the carbon width could be uniquely determined. This was reached by a partial wave analysis of the obtained data. Supplemented were the measurements of the elastic scattering by measurements of the excitation function of the θ 0 exit channel of the reaction 12 ( 12 C, θ) 20 Ne in the energy range from 5.2 ≤ E c.m. ≤ 6.1 MeV. At some known resonances in the 16 O+ 16 O system in the energy range from 15.5 MeV to 18 MeV the spin and the elastic width of these structures could be determined. For this a constrained partial wave analysis on complete angular distributions of the elastic scattering (9 0 ≤ θ c.m. ≤ 90 0 ) which were measured in energy steps of ΔE c.m. = 50 keV was performed. In the 16 O+ 16 O system besides the proof for the existence of a nuclear glory scattering was given. (orig.) [de

Limiting efficiency of generalized realistic c-Si solar cells coupled to ideal up-converters

Science.gov (United States)

Johnson, Craig M.; Conibeer, Gavin J.

2012-11-01

The detailed balance model of photovoltaic up-conversion is revised for the specific case of a c-Si solar cell under the AM1.5G solar spectrum. The limiting efficiency of an ideal solar cell with a band gap of 1.117 eV may be increased from approximately 33% to 40% with ideal up-conversion. However, real solar cells do not demonstrate the step-function absorption characteristic assumed in the standard detailed balance model. Here, we use tabulated Si refractive index data to develop a generalized model of a realistic conventional c-Si solar cell. The model incorporates optical design and material parameters such as free carrier absorption that have a non-trivial impact on the operation of the up-conversion layer. While these modifications are shown to decrease the absolute limiting efficiency, the benefit of up-conversion is shown to be relatively greater.

Transformation of mouse embryo (C3H 10T1/2) cells by alpha particles

International Nuclear Information System (INIS)

Lloyd, E.L.; Gemmell, A.; Henning, C.B.; Gemmell, D.S.; Zabransky, B.J.

1977-01-01

Mammalian cells in culture (C3H mouse 10T1/2 cells) have been shown here for the first time to be transformed by alpha irradiation when cells were irradiated with 5.6 MeV alpha particles from a Tandem Van de Graaff machine. Malignant tumors were induced following inoculation of the transformed cells into syngeneic hosts. Unirradiated control cells injected at the same concentration have, so far, failed to produce tumors. The morphology of the transformed foci was remarkably similar to that obtained by x rays and chemicals but different from virally transformed cells. When the cells were seeded at low density in the exponential growth phase, the transformation frequency per surviving cell increased approximately as the cube of the dose and peaked at an alpha particle fluence between 1.5 and 2.5 x 10 7 alpha particles per cm 2 (205 to 342 rads). The frequency of the transformation was found to be greatly dependent on the number of cells per dish irradiated. Irradiation of larger numbers resulted in much lower frequencies of transformation. The maximum transformation frequency observed in nine separate experiments was 4 percent of the surviving cells. At doses greater than 200 rads the transformation frequency per surviving cell remained constant. The present results permit us to conclude that alpha irradiation may, indeed, be able to exert a direct effect on the genome of the cell to produce malignancy without any external immunological or hormonal influences

Cell density dependence of transformation frequencies in C3H10T1/2 cells exposed to X-rays

Energy Technology Data Exchange (ETDEWEB)

Bettega, D; Calzolari, P; Ottolenghi, A; Lombardi, L T [Milan Univ. (Italy). Ist. di Fisica; Rimoldi, E [Milan Univ. (Italy). Ist. di Radiologia Veterinaria

1989-12-01

The effects of cell density on transformation frequencies were studied in C3H10T1/2 cells exposed to 0.5 and 7 Gy of 200 kVp X-rays. Initial cell density strongly influenced transformation frequency; this decreased by a factor of between 4 and 10 when the initial seeding density was changed from 50 to 2500 cells/10 cm diameter Petri dish. The data were fitted with two equations: (a) an allometric function represented on a log-log scale by a straight line and (b) a sigmoidal function with plateaux between 50 and 250 cells/dish and above 600. The two curves are compared and their probabilities discussed. Our data indicate that the region between 50 and 250 cells/dish would be the most suitable region for dose-effect measurements. A study of the growth curves at 0.5 and 8.5 Gy shows that cell growth rates are not influenced by initial cell density. (author).

The (gas + liquid) critical properties and phase behaviour of some binary alkanol (C2-C5) + alkane (C5-C12) mixtures

International Nuclear Information System (INIS)

Morton, David W.; Lui, Matthew P.W.; Young, Colin L.

2003-01-01

Previously, the investigation of the (gas + liquid) critical properties of (alkanol + alkane) mixtures has focussed on (primary alkanol + straight chain alkane) mixtures. The experimental data available for (alkanol + alkane) mixtures, which include secondary or tertiary alcohols and/or branched chain alkanes, are extremely limited. This work extends the existing body of data on (alkanol + alkane) mixtures to include mixtures containing these components. Here the (gas + liquid) critical temperatures of 29 {alkanol (C 2 -C 5 ) + alkane (C 5 -C 12 )} mixtures are reported. All the (gas + liquid) critical lines for the binary mixtures studied are continuous, indicating they obey either Type I or Type II phase behaviour

Up-regulation of granzyme B and perforin by staphylococcal enterotoxin C2 mutant induces enhanced cytotoxicity in Hepa1–6 cells

Energy Technology Data Exchange (ETDEWEB)

Zhang, Guojun [Institute of Applied Ecology, Chinese Academy of Sciences, No.72 Wenhua Road Shenhe Dis., Shenyang, Liaoning (China); University of Chinese Academy of Sciences, Beijing (China); Xu, Mingkai, E-mail: mkxu@iae.ac.cn [Institute of Applied Ecology, Chinese Academy of Sciences, No.72 Wenhua Road Shenhe Dis., Shenyang, Liaoning (China); Zhang, Huiwen [Institute of Applied Ecology, Chinese Academy of Sciences, No.72 Wenhua Road Shenhe Dis., Shenyang, Liaoning (China); Song, Yubo [Institute of Applied Ecology, Chinese Academy of Sciences, No.72 Wenhua Road Shenhe Dis., Shenyang, Liaoning (China); University of Chinese Academy of Sciences, Beijing (China); Wang, Jian; Zhang, Chenggang [Institute of Applied Ecology, Chinese Academy of Sciences, No.72 Wenhua Road Shenhe Dis., Shenyang, Liaoning (China)

2016-12-15

Staphylococcal enterotoxin C2 (SEC2), a member of bacterial superantigen, is one of the most potent known activators of T lymphocytes. With this property, SEC2 has already been used in clinic as a tumor immunotherapy agent in China. To increase the antitumor activity, a SEC2 mutant named ST-4 (GKVTG102-106WWH) with amino acid substitutions in T cell receptor (TCR)-binding domain was generated by site-directed mutagenesis, and the molecular mechanism of the enhanced antitumor activity was investigated. Results showed that ST-4 could activate much more Vβ 8.2 and 8.3 T cells and NK cells compared with SEC2, and exhibited significantly enhanced immunocyte stimulation and antitumor activity in vitro. The synthetic peptide sequencing the residues of mutant TCR-binding domain could competitively inhibit the immunocyte stimulation activity of ST-4. Most importantly, ST-4 up-regulated granzyme B and perforin at both mRNA and protein levels. We also found that expression of proapoptotic proteins cytochrome c, BAX and activation of caspase-3, 9 was up-regulated, and antiapoptotic protein Bcl-xL was down-regulated in the treatment with either ST-4 or SEC2. When granzyme B inhibitor or perforin inhibitor is presented, tumor cell viability was significantly rescued. Taken together, we demonstrate that increased ST-4-TCR recognition contributed to massive T cells and NK cells activation. These activated cells released up-regulated granzyme B and perforin, which induced the enhanced tumor cells apoptosis by mitochondrial apoptotic pathway, and ultimately led to enhanced tumor cell growth inhibition. ST-4 may be a promising candidate for antitumor clinic usage in future. - Highlights: • We obtained a SEC2 mutant ST-4 with enhanced superantigen and antitumor activity. • Increased ST-4-TCR recognition contributed to massive T cells and NK cells activation. • Up-regulated GzmB and PRF1 in T cell by ST-4 induced enhanced tumor cells apoptosis. • Enhanced tumor cell apoptosis

Angular correlation measurements for {sup 12}C{sup 12}C,{sup 12}C{sup 12}C 3{sup -} scattering

Energy Technology Data Exchange (ETDEWEB)

Wuosmaa, A.H.; Betts, R.R.; Freer, M.

1995-08-01

Previous studies of inelastic {sup 12}C + {sup 12}C scattering to a variety of final states identified significant resonance behavior in a number of different reaction channels. These resonances can be interpreted as either potential scattering resonances, or as population of cluster structures in the compound nucleus {sup 24}Mg, or as some interplay between the two mechanisms. Currently, for many of these resonances the situation remains unclear. One example is a large peak observed in the excitation function for the 3{sup -} - g.s. excitation, identified in previous work performed at the Daresbury Laboratory in England. This peak is observed at the same center-of-mass energy as one observed in the O{sub 2}{sup +}-O{sub 2}{sup +} inelastic scattering channel. That structure was suggested to correspond to exotic deformed configurations in the compound nucleus {sup 24}Mg. As the peak in the 3{sup -} + g.s. exit channel occurs at precisely the same energy as the purported resonance, it is tempting to associate the two. Before such an association can be confirmed or ruled out, further information must be obtained about the 3{sup -} + g.s. structure. In particular, it is important to determine the angular momenta that dominate the 3{sup -} + g.s. structure.

Radiation-induced genetic instability: no association with changes in radiosensitivity or cell cycle checkpoints in C3H 10T1/2 mouse fibroblasts

International Nuclear Information System (INIS)

Crompton, N.E.A.; Emery, G.C.; Shi Yuquan; Sigg, M.; Blattmann, H.

1998-01-01

We investigated various phenotypic characteristics of radiation-induced morphologically transformed C3H 10T1/2 mouse fibroblasts. The cells were treated with 8 Gy x-rays, and type II/III foci were isolated. Cell lines were developed from these foci, and subsequently clones were established from these focal lines. The clones were examined for DNA content, radiosensitivity and inducible cell cycle arrests. Besides the morphological changes associated with the transformed state, the major difference between the isolated focal lines or derived clones and the parental C3H 10T1/2 line was one of ploidy. The transformed cells often displayed aneuploid and multiple polyploid populations. No change in the radiosensitivity of the transformed cells was observed. Furthermore, the two major radiation- and staurosporine-induced G1 and G2 cell cycle arrests observed in the parental cell line were also observed in the morphological transformants, suggesting that checkpoint function was normal. (orig.)

Kinetics of C3H 10T1/2 mouse cells

International Nuclear Information System (INIS)

Metting, N.F.; Mortenson, E.; Braby, L.A.; Nelson, J.M.

1987-01-01

The 10T1/2 cell line has been brought to PNL because of its usefulness as an in vitro biological system for transformation assay, and because a fast repair process in response to irradiation has been observed in these cells. Before using the cells in radiobiological studies, it was important to observe the growth kinetics of our cultures and verify their similarity to those reported by other laboratories in which they are used, so that our subsequent measurements could be compared with and added to the body of scientific work already compiled. To become familiar with the cell morphology and cloning characteristics, we conducted an experiment in which a number of flasks were seeded with about 350 cells each. Each day, starting with the second day after plating, a flask was stained and the number of clones plus the number of cells in each clone were counted. There are several types of cell morphologies within the population. Flow cytometry (FCM) studies were used to assess cell cycle characteristics, Again, a number of flasks were seeded, but this time with a large number of exponentially growing cells. Measurements indicated that the cultures were probably more or less synchronized at their initiation by plating efficiency effects, that they maintained a loose synchrony by phases through about the eighth day and that before at least the eleventh day they began to pile up in G1 phase

One-pot synthesis of 4,8-dibromobenzo[1,2-c;4,5-c']bis[1,2,5]thiadiazole.

Science.gov (United States)

Tam, Teck Lip; Li, Hairong; Wei, Fengxia; Tan, Ke Jie; Kloc, Christian; Lam, Yeng Ming; Mhaisalkar, Subodh G; Grimsdale, Andrew C

2010-08-06

A one-step synthesis of 4,8-dibromobenzo[1,2-c;4,5-c']bis[1,2,5]thiadiazole with use of 1,2,4,5-tetraaminobenzene tetrahydrobromide and thionyl bromide in good yield is reported. This unit can then be used in the synthesis of low bandgap materials via palladium-catalyzed coupling reactions. The approach offers a quick and easy way to prepare low bandgap materials as compared to the current literature methods.

Double-step processes in the 12C(p,d)11C reaction at 45 MeV

International Nuclear Information System (INIS)

Couvert, Pierre.

1974-01-01

12 C(p,d) 11 C pick-up reaction was performed with a 45 MeV proton beam. A 130keV energy resolution was obtained and angular distributions of nine of the ten first levels of 11 C have been extracted within a large angular range. Assuming only neutron direct transfert, the strong relative excitation of high spin levels cannot be reproduced by a DWBA analysis. The double-step process assumption seems to be verified by a systematical analysis of the (p,d) reaction mechanisms. This analysis is done in the coupled-channel formalism for the five first negative parity states of 11 C. The 3/2 - ground state is essentially populated by the direct transfer of a Psub(3/2) neutron. The contribution of a double-step process, via the 2 + inelastic excitation of 12 C, is important for the four other states. A mechanism which assumes a deuteron inelastic scattering on the 11 C final nucleus after the neutron transfer cannot be neglected and improves the fits when it is taken into account [fr

C5a regulates IL-12+ DC migration to induce pathogenic Th1 and Th17 cells in sepsis.

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Ning Ma

Full Text Available OBJECTIVE: It is well known that complement system C5a is excessively activated during the onset of sepsis. However, it is unclear whether C5a can regulate dentritic cells (DCs to stimulate adaptive immune cells such as Th1 and Th17 in sepsis. METHODS: Sepsis was induced by cecal ligation and puncture (CLP. CLP-induced sepsis was treated with anti-C5a or IL-12. IL-12(+DC, IFNγ(+Th1, and IL-17(+Th17 cells were analyzed by flow cytometry. IL-12 was measured by ELISA. RESULTS: Our studies here showed that C5a induced IL-12(+DC cell migration from the peritoneal cavity to peripheral blood and lymph nodes. Furthermore, IL-12(+DC cells induced the expansion of pathogenic IFNγ(+Th1 and IL-17(+Th17 cells in peripheral blood and lymph nodes. Moreover, IL-12, secreted by DC cells in the peritoneal cavity, is an important factor that prevents the development of sepsis. CONCLUSION: Our data suggests that C5a regulates IL-12(+DC cell migration to induce pathogenic Th1 and Th17 cells in sepsis.

Pycnonuclear 12C+12C reaction at zero temperature

International Nuclear Information System (INIS)

Gasques, L R; Beard, M; Chamon, L C; Wiescher, M

2005-01-01

We present pycnonuclear reaction calculations for a one-component ionic crystal at zero temperature considering different theoretical approaches. The rates depend directly on the determination of the astrophysical S-factor at low energies, which has been obtained through the barrier penetration formalism. A totally parameter-free model for the real part of the nuclear interaction has been employed in the calculation of 12 C+ 12 C fusion cross sections

The {sup 12}C/{sup 13}C Ratio in Sgr B2(N): Constraints for Galactic Chemical Evolution and Isotopic Chemistry

Energy Technology Data Exchange (ETDEWEB)

Halfen, D. T.; Ziurys, L. M. [Department of Chemistry and Biochemistry, University of Arizona, 1305 E. 4th Street, Tucson, AZ 85719 (United States); Woolf, N. J., E-mail: halfend@email.arizona.edu [Department of Astronomy, Arizona Radio Observatory, University of Arizona, 933 North Cherry Avenue, Tucson, AZ 85721 (United States)

2017-08-20

A study has been conducted of {sup 12}C/{sup 13}C ratios in five complex molecules in the Galactic center. H{sub 2}CS, CH{sub 3}CCH, NH{sub 2}CHO, CH{sub 2}CHCN, and CH{sub 3}CH{sub 2}CN and their {sup 13}C-substituted species have been observed in numerous transitions at 1, 2, and 3 mm, acquired in a spectral-line survey of Sgr B2(N), conducted with the telescopes of the Arizona Radio Observatory (ARO). Between 22 and 54 individual, unblended lines for the {sup 12}C species and 2–54 for {sup 13}C-substituted analogs were modeled in a global radiative transfer analysis. All five molecules were found to consistently exhibit two velocity components near V {sub LSR} ∼ 64 and 73 km s{sup −1}, with column densities ranging from N {sub tot} ∼ 3 × 10{sup 14} − 4 × 10{sup 17} cm{sup −2} and ∼2 × 10{sup 13} − 1 × 10{sup 17} cm{sup −2} for the {sup 12}C and {sup 13}C species, respectively. Based on 14 different isotopic combinations, ratios were obtained in the range {sup 12}C/{sup 13}C = 15 ± 5 to 33 ± 13, with an average value of 24 ± 7, based on comparison of column densities. These measurements better anchor the {sup 12}C/{sup 13}C ratio at the Galactic center, and suggest a slightly revised isotope gradient of {sup 12}C/{sup 13}C = 5.21(0.52) D {sub GC} + 22.6(3.3). As indicated by the column densities, no preferential {sup 13}C enrichment was found on the differing carbon sites of CH{sub 3}CCH, CH{sub 2}CHCN, and CH{sub 3}CH{sub 2}CN. Because of the elevated temperatures in Sgr B2(N), {sup 13}C isotopic substitution is effectively “scrambled,” diminishing chemical fractionation effects. The resulting ratios thus reflect stellar nucleosynthesis and Galactic chemical evolution, as is likely the case for most warm clouds.

Inelastic 12C(12C, 12C)12C scattering via the Glauber optical potential at E/A=30 MeV and 85 MeV

International Nuclear Information System (INIS)

Chauvin, J.; Lebrun, D.; Durand, F.; Buenerd, M.

1984-07-01

It is shown that some approximations implied in the optical limit of the Glauber theory are justified for low energies heavy-ions surface collisions. The Glauber optical potential -with no free parameters- leads to reasonable predictions for both elastic and inelastic scattering in the 12 C + 12 C system

The 12C/ 13C isotopic ratio in Titan hydrocarbons from Cassini/CIRS infrared spectra

Science.gov (United States)

Nixon, C. A.; Achterberg, R. K.; Vinatier, S.; Bézard, B.; Coustenis, A.; Irwin, P. G. J.; Teanby, N. A.; de Kok, R.; Romani, P. N.; Jennings, D. E.; Bjoraker, G. L.; Flasar, F. M.

2008-06-01

We have analyzed infrared spectra of Titan recorded by the Cassini Composite Infrared Spectrometer (CIRS) to measure the isotopic ratio 12C/ 13C in each of three chemical species in Titan's stratosphere: CH 4, C 2H 2 and C 2H 6. This is the first measurement of 12C/ 13C in any C 2 molecule on Titan, and the first measurement of 12CH 4/ 13CH 4 (non-deuterated) on Titan by remote sensing. Our spectra cover five widely-spaced latitudes, 65° S to 71° N and we have searched for both latitude variability of 12C/ 13C within a given species, and also for differences between the 12C/ 13C in the three gases. For CH 4 alone, we find C12/C13=76.6±2.7 (1- σ), essentially in agreement with the 12CH 4/ 13CH 4 measured by the Huygens Gas Chromatograph/Mass Spectrometer instrument (GCMS) [Niemann, H.B., and 17 colleagues, 2005. Nature 438, 779-784]: 82.3±1.0, and also with measured values in H 13CN and 13CH 3D by CIRS at lower precision [Bézard, B., Nixon, C., Kleiner, I., Jennings, D., 2007. Icarus 191, 397-400; Vinatier, S., Bézard, B., Nixon, C., 2007. Icarus 191, 712-721]. For the C 2 species, we find C12/C13=84.8±3.2 in C 2H 2 and 89.8±7.3 in C 2H 6, a possible trend of increasingly value with molecular mass, although these values are both compatible with the Huygens GCMS value to within error bars. There are no convincing trends in latitude. Combining all fifteen measurements, we obtain a value of C12/C13=80.8±2.0, also compatible with GCMS. Therefore, the evidence is mounting that 12C/ 13C is some 8% lower on Titan than on the Earth (88.9, inorganic standard), and lower than typical for the outer planets ( 88±7 [Sada, P.V., McCabe, G.H., Bjoraker, G.L., Jennings, D.E., Reuter, D.C., 1996. Astrophys. J. 472, 903-907]). There is no current model for this enrichment, and we discuss several mechanisms that may be at work.

Ordering effects in benzo[1,2-b:4,5-b']difuran-thieno[3,4-c]pyrrole-4,6- dione polymers with >7% solar cell efficiency

KAUST Repository

Warnan, Julien; Cabanetos, Clement; El Labban, Abdulrahman; Hansen, Michael Ryan; Tassone, Christopher J.; Toney, Michael F.; Beaujuge, Pierre

2014-01-01

Benzo[1,2-b:4,5-b']difuran-thieno[3,4-c]pyrrole-4,6-dione (PBDFTPD) polymers prepared by microwave-assisted synthesis can achieve power conversion efficiencies (PCEs) >7% in bulk-heterojunction solar cells with phenyl-C61/71-butyric acid methyl

Jaridonin-induced G2/M phase arrest in human esophageal cancer cells is caused by reactive oxygen species-dependent Cdc2-tyr15 phosphorylation via ATM–Chk1/2–Cdc25C pathway

Energy Technology Data Exchange (ETDEWEB)

Ma, Yong-Cheng [Clinical Pharmacology Laboratory, Henan Province People' s Hospital, No. 7, Wei Wu Road, Zhengzhou, Henan (China); Su, Nan [Department of Quality Detection and Management, Henan University of Animal Husbandry and Economy, Zhengzhou, Henan (China); Shi, Xiao-Jing; Zhao, Wen; Ke, Yu [School of Pharmaceutical Sciences, Zhengzhou University, No. 100, Science Avenue, Zhengzhou, Henan (China); Zi, Xiaolin [Department of Urology, University of California, Irvine, Orange, CA (United States); Department of Pharmacology, University of California, Irvine, Orange, CA (United States); Department of Pharmaceutical Sciences, University of California, Irvine, Orange, CA (United States); Zhao, Ning-Min; Qin, Yu-Hua; Zhao, Hong-Wei [Clinical Pharmacology Laboratory, Henan Province People' s Hospital, No. 7, Wei Wu Road, Zhengzhou, Henan (China); Liu, Hong-Min, E-mail: liuhm@zzu.edu.cn [School of Pharmaceutical Sciences, Zhengzhou University, No. 100, Science Avenue, Zhengzhou, Henan (China)

2015-01-15

Jaridonin, a novel diterpenoid from Isodon rubescens, has been shown previously to inhibit proliferation of esophageal squamous cancer cells (ESCC) through G2/M phase cell cycle arrest. However, the involved mechanism is not fully understood. In this study, we found that the cell cycle arrest by Jaridonin was associated with the increased expression of phosphorylation of ATM at Ser1981 and Cdc2 at Tyr15. Jaridonin also resulted in enhanced phosphorylation of Cdc25C via the activation of checkpoint kinases Chk1 and Chk2, as well as in increased phospho-H2A.X (Ser139), which is known to be phosphorylated by ATM in response to DNA damage. Furthermore, Jaridonin-mediated alterations in cell cycle arrest were significantly attenuated in the presence of NAC, implicating the involvement of ROS in Jaridonin's effects. On the other hand, addition of ATM inhibitors reversed Jaridonin-related activation of ATM and Chk1/2 as well as phosphorylation of Cdc25C, Cdc2 and H2A.X and G2/M phase arrest. In conclusion, these findings identified that Jaridonin-induced cell cycle arrest in human esophageal cancer cells is associated with ROS-mediated activation of ATM–Chk1/2–Cdc25C pathway. - Highlights: • Jaridonin induced G2/M phase arrest through induction of redox imbalance. • Jaridonin increased the level of ROS through depleting glutathione in cell. • ATM–Chk1/2–Cdc25C were involved in Jaridonin-induced cell cycle arrest. • Jaridonin selectively inhibited cancer cell viability and cell cycle progression.

Neutron electric form factor up to Q2 = 1.47 GeV/c2

International Nuclear Information System (INIS)

Madey, Richard; Semenov, Andrei; Taylor, S.; Aghalaryan, Aram; Crouse, Erick; MacLachlan, Glen; Plaster, Bradley; Shigeyuki Tajima; William Tireman; Chenyu Yan; Abdellah Ahmidouch; Brian Anderson; Hartmuth Arenhovel; Razmik Asaturyan; Baker, O.; Alan Baldwin; Herbert Breuer; Roger Carlini; Christy, E.; Steve Churchwell; Leon Cole; Areg Danagoulian; Donal Day; Mostafa Elaasar; Rolf Ent; Manouchehr Farkhondeh; Howard Fenker; John Finn; Liping Gan; Kenneth Garrow; Paul Gueye; Calvin Howell; Bitao Hu; Mark Jones; James Kelly; Cynthia Keppel; Mahbubul Khandaker; Wooyoung Kim; Stanley Kowalski; Allison Lung; David Mack; Manley, D.; Pete Markowitz; Joseph Mitchell; Hamlet Mkrtchyan; Allena Opper; Charles Perdrisat; Vina Punjabi; Brian Raue; Tilmann Reichelt; Joerg Reinhold; Julie Roche; Yoshinori Sato; Irina Semenova; Wonick Seo; Neven Simicevic; Smith, G.; Samuel Stepanyan; Vardan Tadevosyan; Liguang Tang; Paul Ulmer; William Vulcan; Watson, J. W.; Steven Wells; Frank Wesselmann; Stephen Wood; Chen Yan; Seunghoon Yang; Lulin Yuan; Wei-Ming Zhang; Hong Guo Zhu; Xiaofeng Zhu

2003-01-01

The ratio of the electric to the magnetic form factor of the neutron, g /equiv G En /G Mn , was measured via recoil polarimetry (R.G. Arnold, C.E. Carlson, F. Gross, Phys. Rev. C 23, 363 (1981)) from the quasielastic 2 H (/mathop(e)/limitse' /mathop(n)/limits) 1H reaction at three values of Q 2 (viz, 0.45, 1.15, and 1.47 (GeV/c) 2 ) in Hall C of the Thomas Jefferson National Accelerator Facility. The data reveal that GEn continues to follow the Galster parameterization up to Q 2 = 1.15 (GeV/c) 2 and rises above the Galster parameterization at Q 2 = 1.47 (GeV/c) 2

Differential cross section measurement of elastic scattering 12C(p,p)12C in the astrophysical range of energy

International Nuclear Information System (INIS)

Baktibayev, M.K.; Burminskii, V.P.; Burtebaev, N.; Dzazairov -Kakhramanov, V.; Hassan, S.F.; Satpaev, N.K.; Zazulin, D.M.

2004-01-01

Full text: The fulfillment of planned works on measurements of differential cross sections of elastic scattering of protons on nuclear 12 C at the energy region of 350†1050 keV suggests the preparation of thin self - supporting carbon target. The self - supporting target is necessary in order to perform investigations in the total angular range. In the future last data will be used in order to determine optical potentials and scattering phases for this nuclear in the energy range of astrophysical interest. There was prepared target layer of the 12 C with natural composition of carbon and of thickness of 17.4 μg/cm 2 . The spraying was conducted in the vacuum evaporation installation (VUP - 4) by an electron bombardment method. Carbon was sprayed on a glass plate with previously deposited of layer salt. After a heating during 12 hours at the temperature of 150 o C the film of carbon was floated from glass plate and self - supporting target has been picked up on the specially prepared target frame. In order to determine thickness of target there was used the resonance chamber, installed in the protons channel of the accelerator RAC - 2 - 1 (INP NNC RK), with the help of which there was measured energy loss of the protons beam during the passage through target, disposed in the central chamber. For this purpose there was used the reaction 27 Al(p,γ) 28 Si with narrow resonance with E R = 992 keV and with detection of gamma-quanta with E γ = 1779 keV. On shift of the resonance E R =992 keV in the reaction 27 Al(p,γ) 28 Si, which takes place owing to protons energy loss in the thickness of carbon film, and using table values of brake quantities S(E p )[MeV·cm 2 /g] [1], there was determined thickness of this fine film. Such the method allows to determine thicknesses of films in the interval of (10 † 100) mcg/cm 2 with the accuracy of not worse than 5%. In the present work there were carried out measurements of angular distributions of cross sections of the

The 12C/13C Isotopic Ratio in Planetary Nebulae as Deduced from IUE Data

Science.gov (United States)

Miskey, C. L.; Feibelman, W. A.; Bruhweiler, F. C.

2000-05-01

The relative abundances of C, N, and O and the isotopic ratio of 12C/13C represent tracers of nucleosynthesis in intermediate stars with main-sequence masses between 0.6 and 8.0 solar masses in our Galaxy. Determining these abundances and the isotopic 12C/13C ratio in planetary nebulae (PNe) represent perhaps the best means to discern exactly how the ISM is enriched by CNO stellar nucleosynthesis. Walsh et al. (1996) and Clegg et al. (1997), using the Hubble Space Telescope, have derived the isotopic 12C/13C abundance ratio in the galactic carbon-rich PN, NGC 3918, and placed marginal constraints on it for the Magellanic PNe, N2 (SMC) and N122 (LMC). This was done using the well-known 12C 3P-1S (J=1-0 and J=2-0) transitions of C+2 at 1906.68 Angstroms and 1908.77 Angstroms and a J=0-0 transition at 1909.6 Angstroms, which is strictly forbidden in 12C. The finite nuclear spin of 13C (I=1/2) permits a corresponding F=1/2-1/2 electric dipole transition not seen in 12C. Since the 1909.6 Angstroms line is well separated from the other two 12C transitions, it provides an important means of determining 12C/13C in planetary nebulae. We have just completed a search of archival International Ultraviolet Explorer (IUE) high-dispersion spectra of approximately three dozen PNe, and derived 12C/13C ratios of 39 and 23 for the galactic PNe, NGC 2440 and NGC 6302, respectively. These are values much lower than the solar value of 89. In the other objects, the limited S/N of the IUE data indicate 12C/13C ratio upper limits much higher than 50. The implications of these results and their pertinence to stellar evolution are discussed.

Hypomorphic Smn knockdown C2C12 myoblasts reveal intrinsic defects in myoblast fusion and myotube morphology

International Nuclear Information System (INIS)

Shafey, Dina; Cote, Patrice D.; Kothary, Rashmi

2005-01-01

Dosage of the survival motor neuron (SMN) protein has been directly correlated with the severity of disease in patients diagnosed with spinal muscular atrophy (SMA). It is also clear that SMA is a neurodegenerative disorder characterized by the degeneration of the α-motor neurons in the anterior horn of the spinal cord and atrophy of the associated skeletal muscle. What is more controversial is whether it is neuronal and/or muscle-cell-autonomous defects that are responsible for the disease per se. Although motor neuron degeneration is generally accepted as the primary event in SMA, intrinsic muscle defects in this disease have not been ruled out. To gain a better understanding of the influence of SMN protein dosage in muscle, we have generated a hypomorphic series of myoblast (C2C12) stable cell lines with variable Smn knockdown. We show that depletion of Smn in these cells resulted in a decrease in the number of nuclear 'gems' (gemini of coiled bodies), reduced proliferation with no increase in cell death, defects in myoblast fusion, and malformed myotubes. Importantly, the severity of these abnormalities is directly correlated with the decrease in Smn dosage. Taken together, our work supports the view that there is an intrinsic defect in skeletal muscle cells of SMA patients and that this defect contributes to the overall pathogenesis in this devastating disease

Excitation functions of inelastic and transfer channels in {sup 12} C+{sup 12} C around E{sub c.m.} = 32.5 MeV

Energy Technology Data Exchange (ETDEWEB)

Szilner, S.; Basrak, Z.; Freeman, R.M.; Haas, F.; Beck, C. [Strasbourg-1 Univ., 67 (France). Centre de Recherches Nucleaires]|[Strasbourg-1 Univ., 67 (France); Morsad, A. [Universite Hassan II, Casablanca (Morocco). Faculte des Sciences

1996-12-31

A prominent and wide resonance centered at E{sub c.m.} = 32.5 MeV has recently been found in the (0{sub 2}{sup +}, 0{sub 2}{sup +}) inelastic channel of the {sup 12}C+{sup 12}C reaction. It has been suggested that it corresponds to a 6{alpha}-particle-chain state in {sup 24}Mg. In the present work we study {sup 12}C+{sup 12}C excitation functions between center-of-mass energies of 30 and 35 MeV in steps of 250 keV for weakly populated outgoing channels. We present the inelastic channels to the states above the {alpha}-particle decay threshold, (0{sub 1}{sup +},0{sub 2}{sup +}), (0{sub 1}{sup +},3{sub 1}{sup -}), and (0{sub 1}{sup +},4{sub 1}{sup 4}), and the one- and two-nucleon transfer channels. In the inelastic and the transfer channels we observe correlated intermediate-width structures at E{sub c.m.} = 31, 32.5, and 33.5 MeV, whose widths are appreciably smaller than the width measured in the (0{sub 2}{sup +},0{sub 2}{sup +}) channel. Our E{sub c.m.} = 31, 32.5, and 33.5 MeV, whose widths are appreciably smaller than the width measured in the (0{sub 2}{sup +},0{sub 2}{sup +}) channel. Our E{sub c.m.} = 32.5 MeV angular distribution of the (0{sub 1}{sup +},0{sub 2}{sup +})channel exhibits oscillatory behavior and, unlike that of the (0{sup +}{sub 2}, 0{sup +}{sub 2}) channel, does not display enhancement around {Theta}{sub c.m.} = 90 deg. Data were collected via the kinematic coincidence technique. For data reduction we use a novel approach allowing for the extraction of results on non-binary channels. (authors). 39 refs.

C30F12.4 influences oogenesis, fat metabolism, and lifespan in C. elegans

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Lu Wang

2016-09-01

Full Text Available ABSTRACT Reproduction, fat metabolism, and longevity are intertwined regulatory axes; recent studies in C. elegans have provided evidence that these processes are directly coupled. However, the mechanisms by which they are coupled and the reproductive signals modulating fat metabolism and lifespan are poorly understood. Here, we find that an oogenesis-enriched gene, c30f12.4, is specifically expressed and located in germ cells and early embryos; when the gene is knocked out, oogenesis is disrupted and brood size is decreased. In addition to the reproductive phenotype, we find that the loss of c30f12.4 alters fat metabolism, resulting in decreased fat storage and smaller lipid droplets. Meanwhile, c30f12.4 mutant worms display a shortened lifespan. Our results highlight an important role for c30f12.4 in regulating reproduction, fat homeostasis, and aging in C. elegans, which helps us to better understand the relationship between these processes.

Suspension state increases reattachment of breast cancer cells by up-regulating lamin A/C.

Science.gov (United States)

Zhang, Xiaomei; Lv, Yonggang

2017-12-01

Extravasation is a rate-limiting step of tumor metastasis, for which adhesion to endothelium of circulating tumor cells (CTCs) is the prerequisite. The suspension state of CTCs undergoing detachment from primary tumor is a persistent biomechanical cue, which potentially regulates the biophysical characteristics and cellular behaviors of tumor cells. In this study, breast tumor cells MDA-MB-231 in suspension culture condition were used to investigate the effect of suspension state on reattachment of CTCs. Our study demonstrated that suspension state significantly increased the adhesion ability of breast tumor cells. In addition, suspension state markedly promoted the formation of stress fibers and focal adhesions and reduced the motility in reattached breast cancer cells. Moreover, lamin A/C was reversibly accumulated at posttranscriptional level under suspension state, improving the cell stiffness of reattached breast cancer cells. Disruption of actin cytoskeleton by cytochalasin D caused lamin A/C accumulation. Conversely, decreasing actomyosin contraction by ROCK inhibitor Y27632 reduced lamin A/C level. Knocking down lamin A/C weakened the suspension-induced increase of adhesion, and also abolished the suspension-induced decrease of motility and increase of stress fibers and focal adhesion in reattaching tumor cells, suggesting a crucial role of lamin A/C. In conclusion, it was demonstrated that suspension state promoted the reattachment of breast tumor cells by up-regulating lamin A/C via cytoskeleton disruption. These findings highlight the important role of suspension state for tumor cells in tumor metastasis. Copyright © 2017 Elsevier B.V. All rights reserved.

L-Arginine Enhances Protein Synthesis by Phosphorylating mTOR (Thr 2446 in a Nitric Oxide-Dependent Manner in C2C12 Cells

Directory of Open Access Journals (Sweden)

Ruxia Wang

2018-01-01

Full Text Available Muscle atrophy may arise from many factors such as inactivity, malnutrition, and inflammation. In the present study, we investigated the stimulatory effect of nitric oxide (NO on muscle protein synthesis. Primarily, C2C12 cells were supplied with extra L-arginine (L-Arg in the culture media. L-Arg supplementation increased the activity of inducible nitric oxide synthase (iNOS, the rate of protein synthesis, and the phosphorylation of mTOR (Thr 2446 and p70S6K (Thr 389. L-NAME, an NOS inhibitor, decreased NO concentrations within cells and abolished the stimulatory effect of L-Arg on protein synthesis and the phosphorylation of mTOR and p70S6K. In contrast, SNP (sodium nitroprusside, an NO donor, increased NO concentrations, enhanced protein synthesis, and upregulated mTOR and p70S6K phosphorylation, regardless of L-NAME treatment. Blocking mTOR with rapamycin abolished the stimulatory effect of both L-Arg and SNP on protein synthesis and p70S6K phosphorylation. These results indicate that L-Arg stimulates protein synthesis via the activation of the mTOR (Thr 2446/p70S6K signaling pathway in an NO-dependent manner.

Sub-Coulomb α Transfers on C12 and the C12(α, γ)O16 S Factor

International Nuclear Information System (INIS)

Brune, C. R.; Geist, W. H.; Kavanagh, R. W.; Veal, K. D.

1999-01-01

The 12 C( α, γ) 16 O reaction is crucial for the understanding of He burning in massive stars, but the low-energy cross section is highly uncertain. To address this problem we have measured at sub-Coulomb energies total cross sections for the 12 C( 6 Li, d) 16 O and 12 C( 7 Li, t) 16 O reactions to the bound 2 + and 1 - states of 16 O . The data are analyzed to obtain the reduced α widths of these states. Together with capture and phase-shift data, these results provide for a more accurate determination of the low-energy 12C(α, γ) 16 O S factor: S E1 (0.3 MeV) =101±17 keV b and S E2 (0.3 MeV) =42 +16 -23 keV b for the E1 and E2 multipole components of the reaction. (c) 1999 The American Physical Society

Fibroblast growth factor-2 up-regulates the expression of nestin through the Ras–Raf–ERK–Sp1 signaling axis in C6 glioma cells

International Nuclear Information System (INIS)

Chang, Kai-Wei; Huang, Yuan-Li; Wong, Zong-Ruei; Su, Peng-Han; Huang, Bu-Miin; Ju, Tsai-Kai; Yang, Hsi-Yuan

2013-01-01

Highlights: •Nestin expression in C6 glioma cells is induced by FGF-2. •Nestin expression is induced by FGF-2 via de novo RNA and protein synthesis. •The FGFR inhibitor SU5402 blocks the FGF-2-induced nestin expression. •The mRNA of FGFR1 and 3 are detected in C6 glioma cells. •Ras–Raf–ERK–Sp1 signaling pathway is responsibe for FGF-2-induced nestin expression. -- Abstract: Nestin is a 240-kDa intermediate filament protein expressed mainly in neural and myogenic stem cells. Although a substantial number of studies have focused on the expression of nestin during development of the central nervous system, little is known about the factors that induce and regulate its expression. Fibroblast growth factor-2 (FGF-2) is an effective mitogen and stimulates the proliferation and differentiation of a subset of nestin-expressing cells, including neural progenitor cells, glial precursor cells, and smooth muscle cells. To assess whether FGF-2 is a potent factor that induces the expression of nestin, C6 glioma cells were used. The results showed that nestin expression was up-regulated by FGF-2 via de novo RNA and protein synthesis. Our RT-PCR results showed that C6 glioma cells express FGFR1/3, and FGFRs is required for FGF-2-induced nestin expression. Further signaling analysis also revealed that FGF-2-induced nestin expression is mediated through FGFR–MAPK–ERK signaling axis and the transcriptional factor Sp1. These findings provide new insight into the regulation of nestin in glial system and enable the further studies on the function of nestin in glial cells

Fibroblast growth factor-2 up-regulates the expression of nestin through the Ras–Raf–ERK–Sp1 signaling axis in C6 glioma cells

Energy Technology Data Exchange (ETDEWEB)

Chang, Kai-Wei [Institute of Molecular and Cellular Biology, National Taiwan University, Taipei 106, Taiwan (China); Huang, Yuan-Li [Department of Biotechnology, College of Health Science, Asia University, Taichung 413, Taiwan (China); Wong, Zong-Ruei; Su, Peng-Han [Institute of Molecular and Cellular Biology, National Taiwan University, Taipei 106, Taiwan (China); Huang, Bu-Miin [Department of Cell Biology and Anatomy, National Cheng-Kung University, Tainan 701, Taiwan (China); Ju, Tsai-Kai [Instrumentation Center, National Taiwan University, Taipei 106, Taiwan (China); Technology Commons, College of Life Science, National Taiwan University, Taipei 106, Taiwan (China); Yang, Hsi-Yuan, E-mail: hyhy@ntu.edu.tw [Institute of Molecular and Cellular Biology, National Taiwan University, Taipei 106, Taiwan (China)

2013-05-17

Highlights: •Nestin expression in C6 glioma cells is induced by FGF-2. •Nestin expression is induced by FGF-2 via de novo RNA and protein synthesis. •The FGFR inhibitor SU5402 blocks the FGF-2-induced nestin expression. •The mRNA of FGFR1 and 3 are detected in C6 glioma cells. •Ras–Raf–ERK–Sp1 signaling pathway is responsibe for FGF-2-induced nestin expression. -- Abstract: Nestin is a 240-kDa intermediate filament protein expressed mainly in neural and myogenic stem cells. Although a substantial number of studies have focused on the expression of nestin during development of the central nervous system, little is known about the factors that induce and regulate its expression. Fibroblast growth factor-2 (FGF-2) is an effective mitogen and stimulates the proliferation and differentiation of a subset of nestin-expressing cells, including neural progenitor cells, glial precursor cells, and smooth muscle cells. To assess whether FGF-2 is a potent factor that induces the expression of nestin, C6 glioma cells were used. The results showed that nestin expression was up-regulated by FGF-2 via de novo RNA and protein synthesis. Our RT-PCR results showed that C6 glioma cells express FGFR1/3, and FGFRs is required for FGF-2-induced nestin expression. Further signaling analysis also revealed that FGF-2-induced nestin expression is mediated through FGFR–MAPK–ERK signaling axis and the transcriptional factor Sp1. These findings provide new insight into the regulation of nestin in glial system and enable the further studies on the function of nestin in glial cells.

Amino acids and insulin act additively to regulate components of the ubiquitin-proteasome pathway in C2C12 myotubes

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Lomax Michael A

2007-03-01

Full Text Available Abstract Background The ubiquitin-proteasome system is the predominant pathway for myofibrillar proteolysis but a previous study in C2C12 myotubes only observed alterations in lysosome-dependent proteolysis in response to complete starvation of amino acids or leucine from the media. Here, we determined the interaction between insulin and amino acids in the regulation of myotube proteolysis Results Incubation of C2C12 myotubes with 0.2 × physiological amino acids concentration (0.2 × PC AA, relative to 1.0 × PC AA, significantly increased total proteolysis and the expression of 14-kDa E2 ubiquitin conjugating enzyme (p Conclusion In a C2C12 myotube model of myofibrillar protein turnover, amino acid limitation increases proteolysis in a ubiquitin-proteasome-dependent manner. Increasing amino acids or leucine alone, act additively with insulin to down regulate proteolysis and expression of components of ubiquitin-proteasome pathway. The effects of amino acids on proteolysis but not insulin and leucine, are blocked by inhibition of the mTOR signalling pathway.

CHARACTERIZATION OF P2-PURINOCEPTOR MEDIATED CYCLIC-AMP FORMATION IN MOUSE C2C12 MYOTUBES

NARCIS (Netherlands)

HENNING, RH; DUIN, M; DENHERTOG, A; NELEMANS, A

1 The formation of adenosine 3':5'-cyclic monophosphate (cyclic AMP) and inositol(1,4,5)trisphosphate (Ins(1,4,5)P3), induced by ATP and other nucleotides was investigated in mouse C2Cl2 myotubes. 2 ATP (100 muM) and ATPgammaS (100 muM) caused a sustained increase in cyclic AMP content of the cells,

Oxidative stress-induced metabolic changes in mouse C2C12 myotubes studied with high-resolution 13C, 1H, and 31P NMR spectroscopy

DEFF Research Database (Denmark)

Straadt, Ida K; Young, Jette F; Petersen, Bent O

2010-01-01

In this study, stress in relation to slaughter was investigated in a model system by the use of (13)C, (1)H, and (31)P nuclear magnetic resonance (NMR) spectroscopy for elucidating changes in the metabolites in C2C12 myotubes exposed to H(2)O(2)-induced stress. Oxidative stress resulted in lower...... to lower levels of the unlabeled ((12)C) lactate were identified in the (1)H spectra after stress exposure. These data indicate an increase in de novo synthesis of alanine, concomitant with a release of lactate from the myotubes to the medium at oxidative stress conditions. The changes in the metabolite...

Chromosomal aberrations in the bone marrow cells of mice induced by accelerated {sup 12}C{sup 6+} ions

Energy Technology Data Exchange (ETDEWEB)

Ma Xiaofei [Department of Heavy Ion Radiation Biology and Medicine, Institute of Modern Physics, Chinese Academy of Sciences, Lanzhou 730000 (China); School of Nuclear Science and Technology, Lanzhou University, Lanzhou 730000 (China); Key Laboratory of Heavy Ion Radiation Medicine of Chinese Academy of Sciences, Lanzhou 730000 (China); Key Laboratory of Heavy Ion Radiation Medicine of Gansu Province, Lanzhou 730000 (China); Graduate University of Chinese Academy of Sciences, Beijing 100049 (China); Zhang Hong, E-mail: zhangh@impac.ac.cn [Department of Heavy Ion Radiation Biology and Medicine, Institute of Modern Physics, Chinese Academy of Sciences, Lanzhou 730000 (China); Key Laboratory of Heavy Ion Radiation Medicine of Chinese Academy of Sciences, Lanzhou 730000 (China); Key Laboratory of Heavy Ion Radiation Medicine of Gansu Province, Lanzhou 730000 (China); Wang Zhenhua; Min Xianhua; Liu Yang; Wu Zhenhua [Department of Heavy Ion Radiation Biology and Medicine, Institute of Modern Physics, Chinese Academy of Sciences, Lanzhou 730000 (China); Key Laboratory of Heavy Ion Radiation Medicine of Chinese Academy of Sciences, Lanzhou 730000 (China); Key Laboratory of Heavy Ion Radiation Medicine of Gansu Province, Lanzhou 730000 (China); Sun Chao [Department of Heavy Ion Radiation Biology and Medicine, Institute of Modern Physics, Chinese Academy of Sciences, Lanzhou 730000 (China); Key Laboratory of Heavy Ion Radiation Medicine of Chinese Academy of Sciences, Lanzhou 730000 (China); Key Laboratory of Heavy Ion Radiation Medicine of Gansu Province, Lanzhou 730000 (China); Graduate University of Chinese Academy of Sciences, Beijing 100049 (China); Hu Bitao [School of Nuclear Science and Technology, Lanzhou University, Lanzhou 730000 (China)

2011-11-01

Highlights: {yields} 220 MeV/u {sup 12}C{sup 6+} ions is 1.5 times more effective than X-rays in inducing chromosomal aberration in bone marrow cell. {yields} The ratio of dose averaged liner energy transfer is approach the RBE. {yields} {sup 12}C{sup 6+} ions could induce severe mitosis delay. {yields} The cell cycle is not recovered 72 h following irradiation. - Abstract: The whole bodies of 6-week-old male Kun-Ming mice were exposed to different doses of {sup 12}C{sup 6+} ions or X-rays. Chromosomal aberrations of the bone marrow (gaps, terminal deletions and breaks, fragments, inter-chromosomal fusions and sister-chromatid union) were scored in metaphase 9 h after exposure, corresponding to cells exposed in the G{sub 2}-phase of the first mitosis cycle. Dose-response relationships for the frequency of chromosomal aberrations were plotted both by linear and linear-quadratic equations. The data showed that there was a dose-related increase in the frequency of chromosomal aberrations in all treated groups compared to controls. Linear-quadratic equations were a good fit for both radiation types. The compound theory of dual radiation action was applied to decipher the bigger curvature (D{sup 2}) of the dose-response curves of X-rays compared to those of {sup 12}C{sup 6+} ions. Different distributions of the five types of aberrations and different degrees of homogeneity were found between {sup 12}C{sup 6+} ion and X-ray irradiation and the possible underlying mechanism for these phenomena were analyzed according to the differences in the spatial energy deposition of both types of radiation.

Synergistic inhibition of the APC/C by the removal of APC15 in HCT116 cells lacking UBE2C

DEFF Research Database (Denmark)

Garvanska, Dimitriya H; Larsen, Marie Sofie Yoo; Nilsson, Jakob

2016-01-01

that has been shown to depend on the APC/C E2 enzymes, UBE2C and UBE2S. Here we investigate the in vivo role of the APC/C E2 enzymes in SAC silencing using CRISPR/Cas9 genetically engineered HCT116 UBE2C or UBE2S null cell lines. Using live cell assays, we show that UBE2C and UBE2S make a minor...... contribution to SAC silencing in HCT116 cells. Strikingly in cells specifically lacking UBE2C, we observe a strong synergistic inhibition of mitotic progression when we stabilize the MCC on the APC/C by depleting APC15, potentially reflecting increased competition between the MCC and the remaining initiating E...

Atmospheric histories and growth trends of C4F10, C5F12, C6F14, C7F16 and C8F18

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R. F. Weiss

2012-05-01

Full Text Available Atmospheric observations and trends are presented for the high molecular weight perfluorocarbons (PFCs: decafluorobutane (C4F10, dodecafluoropentane (C5F12, tetradecafluorohexane (C6F14, hexadecafluoroheptane (C7F16 and octadecafluorooctane (C8F18. Their atmospheric histories are based on measurements of 36 Northern Hemisphere and 46 Southern Hemisphere archived air samples collected between 1973 to 2011 using the Advanced Global Atmospheric Gases Experiment (AGAGE "Medusa" preconcentration gas chromatography-mass spectrometry systems. A new calibration scale was prepared for each PFC, with estimated accuracies of 6.8% for C4F10, 7.8% for C5F12, 4.0% for C6F14, 6.6% for C7F16 and 7.9% for C8F18. Based on our observations the 2011 globally averaged dry air mole fractions of these heavy PFCs are: 0.17 parts-per-trillion (ppt, i.e., parts per 1012 for C4F10, 0.12 ppt for C5F12, 0.27 ppt for C6F14, 0.12 ppt for C7F16 and 0.09 ppt for C8F18. These atmospheric mole fractions combine to contribute to a global average radiative forcing of 0.35 mW m−2, which is 6% of the total anthropogenic PFC radiative forcing (Montzka and Reimann, 2011; Oram et al., 2012. The growth rates of the heavy perfluorocarbons were largest in the late 1990s peaking at 6.2 parts per quadrillion (ppq, i.e., parts per 1015 per year (yr for C4F10, at 5.0 ppq yr−1 for C5F12 and 16.6 ppq yr−1 for C6F14 and in the early 1990s for C7F16 at 4.7 ppq yr−1 and in the mid 1990s for C8F18 at 4.8 ppq yr−1. The 2011 globally averaged mean atmospheric growth rates of these PFCs are subsequently lower at 2.2 ppq yr−1 for C4F10, 1.4 ppq yr−1 for C5F12, 5.0 ppq yr−1 for C6F14, 3.4 ppq yr−1 for C7F16 and 0.9 ppq yr−1 for C8F18. The more recent slowdown in the growth rates suggests that emissions are declining as compared to the 1980s and 1990s.

Ethane's 12C/13C Ratio in Titan: Implications for Methane Replenishment

Science.gov (United States)

Jennings, Donald E.; Nixon, C. A.; Romani, P. N.; Bjoraker, G. L.; Sada, P. V.; Lunsford, A. W.; Boyle, R. J.; Hesman, B. E.; McCabe, G. H.

2009-01-01

As the .main destination of carbon in the destruction of methane in the atmosphere of Titan, ethane provides information about the carbon isotopic composition of the reservoir from which methane is replenished. If the amount of methane entering the atmosphere is presently equal to the amount converted to ethane, the 12C/13C ratio in ethane should be close to the ratio in the reservoir. We have measured the 12C/13C ratio in ethane both with Cassini CIRS(exp 1) and from the ground and find that it is very close to the telluric standard and outer planet values (89), consistent with a primordial origin for the methane reservoir. The lower 12C/13C ratio measured for methane by Huygens GCMS (82.3) can be explained if the conversion of CH4 to CH3 (and C2H6) favors 12C over 13C with a carbon kinetic isotope effect of 1.08. The time required for the atmospheric methane to reach equilibrium, i.e., for replenishment to equal destruction, is approximately 5 methane atmospheric lifetimes.

Cyclic stretch induced miR-146a upregulation delays C2C12 myogenic differentiation through inhibition of Numb

International Nuclear Information System (INIS)

Kuang Wei; Tan Jiali; Duan Yinzhong; Duan Jianmin; Wang Weijian; Jin Fang; Jin Zuolin; Yuan Xiao; Liu Yanpu

2009-01-01

Proliferation and differentiation of muscle stem cells must be tightly regulated by intrinsic and extrinsic signals for effective regeneration and adaptive response. MicroRNAs have been implicated as potent regulators in diverse biological processes at the level of posttranscriptional repression. In this study, we found that miR-146a was significantly upregulated upon a 48-h cyclic stretch of 5% elongation/10cycles/min. Importantly, miR-146 was predicted to base-pair with sequences in the 3' UTR of Numb, which promotes satellite cell differentiation towards muscle cells by inhibiting Notch signaling. Through reporter assay and exogenous expression experiment, we confirmed Numb was inhibited by miR-146a. Inhibition of miR-146a by antago-miR-146a rescued the expression of Numb and facilitated the differentiation of C2C12 at a cost of compromised proliferation. Thus, for the first time, we propose a role of miR-146a in skewing the balance of muscle differentiation and proliferation through inhibiting the expression of Numb.

12C(d,p) 13C reaction at Esub(d) = 30 MeV to the positive-parity states in 13C

International Nuclear Information System (INIS)

Ohnuma, H.; Hoshino, N.; Mikoshiba, O.

1985-07-01

The 12 C(d, p) 13 C reaction has been studied at Esub(d) = 30 MeV. All the known positive-parity states of 13 C below 10 MeV in excitation energy, including the 7/2 + and 9/2 + states, are observed in this reaction. The angular distributions for these positive-parity bound and unbound states are analyzed in CCBA frame work. The 13 C wave functions, which reproduce the resonant and non-resonant scattering of neutrons from 12 C, also give good accounts of the experimentally observed angular distributions and energy spectra of outgoing protons in the 12 C(d, p) 13 C reaction. In most cases the cross section magnitude and the angular distribution shape are primarily determined by the 0 + x j component, even if it is only a small fraction of the total wave function. An exception is the 7/2 + state, where the main contribution comes from the 2 + x dsub(5/2) component. The inclusion of the 4 + state in 12 C and the gsub(9/2) and gsub(7/2) neutron components in the n + 12 C system has very small effects on the low-spin states, but is indispensable for a good fit to the 7/2 + and 9/2 + angular distributions. The transitions to the negative-parity states, 1/2 1 - , 3/2 1 - , 5/2 - , 7/2 - and 1/2 3 - , are also observed experimentally, and analyzed by DWBA. (author)

Schwann Cells Metabolize Extracellular 2′,3′-cAMP to 2′-AMP

Science.gov (United States)

Verrier, Jonathan D.; Kochanek, Patrick M.

2015-01-01

The 3′,5′-cAMP–adenosine pathway (3′,5′-cAMP→5′-AMP→adenosine) and the 2′,3′-cAMP–adenosine pathway (2′,3′-cAMP→2′-AMP/3′-AMP→adenosine) are active in the brain. Oligodendrocytes participate in the brain 2′,3′-cAMP–adenosine pathway via their robust expression of 2′,3′-cyclic nucleotide 3′-phosphodiesterase (CNPase; converts 2′,3′-cAMP to 2′-AMP). Because Schwann cells also express CNPase, it is conceivable that the 2′,3′-cAMP–adenosine pathway exists in the peripheral nervous system. To test this and to compare the 2′,3′-cAMP–adenosine pathway to the 3′,5′-cAMP–adenosine pathway in Schwann cells, we examined the metabolism of 2′,3′-cAMP, 2′-AMP, 3′-AMP, 3′,5′-cAMP, and 5′-AMP in primary rat Schwann cells in culture. Addition of 2′,3′-cAMP (3, 10, and 30 µM) to Schwann cells increased levels of 2′-AMP in the medium from 0.006 ± 0.002 to 21 ± 2, 70 ± 3, and 187 ± 10 nM/µg protein, respectively; in contrast, Schwann cells had little ability to convert 2′,3′-cAMP to 3′-AMP or 3′,5′-cAMP to either 3′-AMP or 5′-AMP. Although Schwann cells slightly converted 2′,3′-cAMP and 2′-AMP to adenosine, they did so at very modest rates (e.g., 5- and 3-fold, respectively, more slowly compared with our previously reported studies in oligodendrocytes). Using transected myelinated rat sciatic nerves in culture medium, we observed a time-related increase in endogenous intracellular 2′,3′-cAMP and extracellular 2′-AMP. These findings indicate that Schwann cells do not have a robust 3′,5′-cAMP–adenosine pathway but do have a 2′,3′-cAMP–adenosine pathway; however, because the pathway mostly involves 2′-AMP formation rather than 3′-AMP, and because the conversion of 2′-AMP to adenosine is slow, metabolism of 2′,3′-cAMP mostly results in the accumulation of 2′-AMP. Accumulation of 2′-AMP in peripheral nerves postinjury could have

Minocycline enhances mitomycin C-induced cytotoxicity through down-regulating ERK1/2-mediated Rad51 expression in human non-small cell lung cancer cells.

Science.gov (United States)

Ko, Jen-Chung; Wang, Tai-Jing; Chang, Po-Yuan; Syu, Jhan-Jhang; Chen, Jyh-Cheng; Chen, Chien-Yu; Jian, Yun-Ting; Jian, Yi-Jun; Zheng, Hao-Yu; Chen, Wen-Ching; Lin, Yun-Wei

2015-10-01

Minocycline is a semisynthetic tetracycline derivative; it has anti-inflammatory and anti-cancer effects distinct from its antimicrobial function. However, the molecular mechanism of minocycline-induced cytotoxicity in non-small cell lung cancer (NSCLC) cells has not been identified. Rad51 plays a central role in homologous recombination and high levels of Rad51 expression are observed in chemo- or radioresistant carcinomas. Our previous studies have shown that the MKK1/2-ERK1/2 signal pathway maintains the expression of Rad51 in NSCLC cells. In this study, minocycline treatment inhibited cell viability and proliferation of two NSCLC cells, A549 and H1975. Treatment with minocycline decreased Rad51 mRNA and protein levels through MKK1/2-ERK1/2 inactivation. Furthermore, expression of constitutively active MKK1 (MKK1-CA) vectors significantly rescued the decreased Rad51 protein and mRNA levels in minocycline-treated NSCLC cells. However, combined treatment with MKK1/2 inhibitor U0126 and minocycline further decreased the Rad51 expression and cell viability of NSCLC cells. Knocking down Rad51 expression by transfection with small interfering RNA of Rad51 enhanced the cytotoxicity and cell growth inhibition of minocycline. Mitomycin C (MMC) is typically used as a first or second line regimen to treat NSCLC. Compared to a single agent alone, MMC combined with minocycline resulted in cytotoxicity and cell growth inhibition synergistically in NSCLC cells, accompanied with reduced activation of phospho-ERK1/2, and reduced Rad51 protein levels. Overexpression of MKK1-CA or Flag-tagged Rad51 could reverse the minocycline and MMC-induced synergistic cytotoxicity. These findings may have implications for the rational design of future drug regimens incorporating minocycline and MMC for the treatment of NSCLC. Copyright © 2015 Elsevier Inc. All rights reserved.

Emission of particles in the 12 C + 12 C fusion

International Nuclear Information System (INIS)

Martinez Q, E.; Aguilera, E.F.; Rosales, P.

2002-01-01

A fusion process analysis of the 12 C + 12 C reaction is done, using the LILITA program. The analysis consisted mainly in varying the value of the Levels density parameter, determining on this way the value of such parameter which reproduces better the contribution of the different channels of fusion-evaporation of particles for this system at different energies. Moreover a comparison with measures done in the Instituto Nacional de Investigaciones Nucleares is realized. (Author)

Application of inverse kinematics to [sup 1]H+[sup 23]Na[yields][sup 12]C+[sup 12]C

Energy Technology Data Exchange (ETDEWEB)

Bennett, S.J. (School of Physics and Space Research, Birmingham Univ. (United Kingdom)); Clarke, N.M. (School of Physics and Space Research, Birmingham Univ. (United Kingdom)); Freer, M. (School of Physics and Space Research, Birmingham Univ. (United Kingdom)); Fulton, B.R. (School of Physics and Space Research, Birmingham Univ. (United Kingdom)); Hall, S.J. (School of Physics and Space Research, Birmingham Univ. (United Kingdom)); Karban, O. (School of Physics and Space Research, Birmingham Univ. (United Kingdom)); Murgatroyd, J.T. (School of Physics and Space Research, Birmingham Univ. (United Kingdom)); Tungate, G. (School of Physics and Space Research, Birmingham Univ. (United Kingdom)); Gyapong, G.J. (Dept. of Physics, York Univ. (United Kingdom)); Jarvis, N.S. (Dept. of Physics, York Univ. (United Kingdom)); Watson, D.L. (Dept. of Physics, York Univ. (United Kingdom)); Rae, W.D.M. (Dept. of Nuclear Physics, Oxford Univ. (United Kingdom)); Smith, A.E. (Dept. of Nuclear; Charissa Collaboration

1993-08-15

Some experimental techniques used in the measurement and analysis of inverse kinematic reactions are described and applied to a kinematically complete study of the [sup 1]H([sup 23]Na, [sup 12]C)[sup 12]C reaction at an incident energy of 180 MeV. An excitation function is obtained for the energy range of 18.5 to 19.2 MeV in [sup 24]Mg*; equivalent to proton energies from 7.1 MeV to 7.8 MeV for the [sup 23]Na(p, [sup 12]C)[sup 12]C reaction. (orig.)

The effects of ultraviolet light on host cell reactivation and plaque size of Herpes simplex virus type 1 in C3H/10T1/2 mouse cells

International Nuclear Information System (INIS)

Montes, J.G.; Taylor, W.D.

1986-01-01

Herpes simplex virus-type 1 (HSV-1) plaque-forming ability and plaque size were measured on (C3H/10T1/2) cell monolayers as functions of pretreatment dose with UV light at different times before inoculation with virus, in order to determine if UV-enhanced reactivation (ER) of UV-irradiated virus, as well as associated phenomena, could be obtained in this cell system. The number of virus plaques observed (i.e. the capacity of the cells to support virus growth) and the size of the plaques were found to increase substantially with pretreatment of the cells with UV light. However, no significant ER was observed. Therefore, the mechanisms responsible for the increases in plaque size and cell capacity seem to be independent of those responsible for ER. In work by others, C3H/10T1/2 cells have been transformed by UV light at doses similar to those used in this study; the absence of ER of UV-irradiated virus in this study indicates that the mechanism underlying ER is not required for transformation. (author)

Serum Amyloid A Induces Toll-Like Receptor 2-Dependent Inflammatory Cytokine Expression and Atrophy in C2C12 Skeletal Muscle Myotubes.

Science.gov (United States)

Passey, Samantha L; Bozinovski, Steven; Vlahos, Ross; Anderson, Gary P; Hansen, Michelle J

2016-01-01

Skeletal muscle wasting is an important comorbidity of Chronic Obstructive Pulmonary Disease (COPD) and is strongly correlated with morbidity and mortality. Patients who experience frequent acute exacerbations of COPD (AECOPD) have more severe muscle wasting and reduced recovery of muscle mass and function after each exacerbation. Serum levels of the pro-inflammatory acute phase protein Serum Amyloid A (SAA) can rise more than 1000-fold in AECOPD and are predictively correlated with exacerbation severity. The direct effects of SAA on skeletal muscle are poorly understood. Here we have examined SAA effects on pro-inflammatory cachectic cytokine expression (IL-6 and TNFα) and atrophy in C2C12 myotubes. SAA increased IL-6 (31-fold) and TNFα (6.5-fold) mRNA levels compared to control untreated cells after 3h of SAA treatment, and increased secreted IL-6 protein at 24h. OxPAPC, a dual TLR2 and TLR4 inhibitor, reduced the response to SAA by approximately 84% compared to SAA alone, and the TLR2 neutralising antibody T2.5 abolished SAA-induced expression of IL-6, indicating that SAA signalling in C2C12 myotubes is primarily via TLR2. SAA also reduced myotube width by 10-13% and induced a 2.5-fold increase in the expression of the muscle atrophy gene Atrogin-1, suggesting direct effects of SAA on muscle wasting. Blocking of TLR2 inhibited the SAA-induced decrease in myotube width and Atrogin-1 gene expression, indicating that SAA induces atrophy through TLR2. These data demonstrate that SAA stimulates a robust pro-inflammatory response in skeletal muscle myotubes via the TLR2-dependent release of IL-6 and TNFα. Furthermore, the observed atrophy effects indicate that SAA could also be directly contributing to the wasting and poor recovery of muscle mass. Therapeutic strategies targeting this SAA-TLR2 axis may therefore ameliorate muscle wasting in AECOPD and a range of other inflammatory conditions associated with loss of muscle mass.

The 12C/13C ratio in stellar atmospheres. VI. Five luminous cool stars

International Nuclear Information System (INIS)

Hinkle, K.H.; Lambert, D.L.; Snell, R.L.

1976-01-01

The isotopic abundance ratio, 12 C/ 13 C, is derived from the CO vibration rotation lines at 1.6 and 2.3 μ for five cool luminous stars by a simple curve-of-growth technique. A new analysis of CN lines at 8000 A is also described for α Sco and α Ori. Results derived independently from CO and CN are in agreement. Final results are 12 C/ 13 C=7 +- 2(α Ori), 12 +- 3(α Sco), 7 +- 3(β Peg), 25 +- 7(chi Cyg), 17 +- 4(α Her), and 7 +- 1.5(α Boo). The α Boo analysis provides a check on the CO curve-of-growth technique; the 12 C/ 13 C ratio from the 2.3 μ CO lines is in good agreement with the previously determined ratio from CN and CH lines

Synthesis and fluorescence of new 3-biphenylpyrrolo[1,2-c]pyrimidines

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Marian-Laurentiu Tatu

2017-07-01

Full Text Available New pyrrolo[1,2-c]pyrimidines derivates having a biphenyl moiety at position 3 have been synthesized by 1,3-dipolar cycloaddition of their corresponding N-ylides with activated alkynes. FTIR, 1H and 13C NMR spectroscopy and elemental analysis have been used to characterize the structures of the new nine pyrrolo[1,2-c]pyrimidine derivates. Absorption and fluorescence spectra have been recorded. The appropriate solvent for the photoluminescence properties of the studied compounds has been found to be chloroform:acetonitrile mixture (1:1. The main spectral features such as molar extinction coefficients (ε, Stokes shifts, quantum yields using quinine sulphate as standard, fluorescence quenching in the presence of benzoquinone and Stern-Volmer constants have been calculated. The substituent effects on intensity of absorption, maximum absorbance wavelengths and fluorescence parameters have been discussed. The highest quantum yield value was found for ethyl 3-(4-biphenylyl-7-(3,4-dimethoxybenzoylpyrrolo[1,2-c]pyrimidine-5-carboxylate (0.55. The obtained results suggest that the studied compounds are promising candidates for future study in order to evaluate their use in practical applications in fluorescent chemical sensors.

C-cells in colloid goiter

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Lima Marcus A.

2003-01-01

Full Text Available PURPOSE: The aim of this investigation was to quantitatively evaluate C-cells in colloid goiters, analyzing 36 thyroids that were obtained through thyroidectomy from 24 patients with goiter and 12 normal glands from adult patients without thyroid disease, which were used as the control group. MATERIAL AND METHODS: On average, 6 different thyroid areas were sampled and labeled by immunohistochemistry with a monoclonal anticalcitonin antibody, utilizing the avidin-biotin-peroxidase complex. C-cells were counted in fields measuring 1 square centimeter, and the mean number of cells per field was then calculated. Data were statistically analyzed using the Mann-Whitney test. RESULTS: In the colloid goiter group, the number of C-cells ranged from 0 to 23 per field, while in normal controls they ranged from 20 to 148 per field. CONCLUSIONS: These results demonstrate a significant decrease of C-cell number in the colloid goiter group compared with control group, indicating that the hyperplastic process is restricted to follicular cells, to the detriment of C-cells, which probably cease to receive trophic stimuli.

From 14C/12C measurements towards radiocarbon dating of ice

NARCIS (Netherlands)

Oerlemans, J.; Wal, R.S.W. van de; Roijen, J.J. van; Raynaud, D.; Borg, K. van der; Jong, A.F.M. de; Lipenkov, V.; Huybrechts, P.

1994-01-01

A dry extraction method of CO2 included in glacier ice adds a contamination equivalent to 1.8 μg modern carbon for a 35 μg C sample. This enables radiocarbon dating by accelerator mass spectrometry of 35 μg C samples to about 25 000 BP. Measured 14C/12C ratios are presented for a part of the Vostok

Take the "C" Train

Science.gov (United States)

Lawton, Rebecca

2008-01-01

In this essay, the author recalls several of her experiences in which she successfully pulled her boats out of river holes by throwing herself to the water as a sea-anchor. She learned this trick from her senior guides at a spring training. Her guides told her, "When you're stuck in a hole, take the "C" train."" "Meaning?" The author asked her…

Overexpression of MIP2, a novel WD-repeat protein, promotes proliferation of H9c2 cells

International Nuclear Information System (INIS)

Wei, Xing; Song, Lan; Jiang, Lei; Wang, Guiliang; Luo, Xinjing; Zhang, Bin; Xiao, Xianzhong

2010-01-01

WD40 repeat proteins have a wide range of diverse biological functions including signal transduction, cell cycle regulation, RNA splicing, and transcription. Myocardial ischemic preconditioning up-regulated protein 2 (MIP2) is a novel member of the WD40 repeat proteins superfamily that contains five WD40 repeats. Little is known about its biological role, and the purpose of this study was to determine the role of MIP2 in regulating cellular proliferation. Transfection and constitutive expression of MIP2 in the rat cardiomyoblast cell line H9c2 results in enhanced growth of those cells as measured by cell number and is proportional to the amount of MIP2 expressed. Overexpression of MIP2 results in a shorter cell cycle, as measured by flow cytometry. Collectively, these data suggest that MIP2 may participate in the progression of cell proliferation in H9c2 cells.

Hydroxylamine enhances glucose uptake in C2C12 skeletal muscle cells through the activation of insulin receptor substrate 1.

Science.gov (United States)

Kimura, Taro; Kato, Eisuke; Machikawa, Tsukasa; Kimura, Shunsuke; Katayama, Shinji; Kawabata, Jun

2014-02-28

Diabetes mellitus is a global disease, and the number of patients with it is increasing. Of various agents for treatment, those that directly act on muscle are currently attracting attention because muscle is one of the main tissues in the human body, and its metabolism is decreased in type II diabetes. In this study, we found that hydroxylamine (HA) enhances glucose uptake in C2C12 myotubes. Analysis of HA's mechanism revealed the involvement of IRS1, PI3K and Akt that is related to the insulin signaling pathway. Further investigation about the activation mechanism of insulin receptor or IRS1 by HA may provide a way to develop a novel anti-diabetic agent alternating to insulin. Copyright © 2014 Elsevier Inc. All rights reserved.

Multiple C-terminal tail Ca(2+)/CaMs regulate Ca(V)1.2 function but do not mediate channel dimerization.

Science.gov (United States)

Kim, Eun Young; Rumpf, Christine H; Van Petegem, Filip; Arant, Ryan J; Findeisen, Felix; Cooley, Elizabeth S; Isacoff, Ehud Y; Minor, Daniel L

2010-12-01

Interactions between voltage-gated calcium channels (Ca(V)s) and calmodulin (CaM) modulate Ca(V) function. In this study, we report the structure of a Ca(2+)/CaM Ca(V)1.2 C-terminal tail complex that contains two PreIQ helices bridged by two Ca(2+)/CaMs and two Ca(2+)/CaM-IQ domain complexes. Sedimentation equilibrium experiments establish that the complex has a 2:1 Ca(2+)/CaM:C-terminal tail stoichiometry and does not form higher order assemblies. Moreover, subunit-counting experiments demonstrate that in live cell membranes Ca(V)1.2s are monomers. Thus, contrary to previous proposals, the crystallographic dimer lacks physiological relevance. Isothermal titration calorimetry and biochemical experiments show that the two Ca(2+)/CaMs in the complex have different properties. Ca(2+)/CaM bound to the PreIQ C-region is labile, whereas Ca(2+)/CaM bound to the IQ domain is not. Furthermore, neither of lobes of apo-CaM interacts strongly with the PreIQ domain. Electrophysiological studies indicate that the PreIQ C-region has a role in calcium-dependent facilitation. Together, the data show that two Ca(2+)/CaMs can bind the Ca(V)1.2 tail simultaneously and indicate a functional role for Ca(2+)/CaM at the C-region site.

Spondylolysis of C-2 in children 3 years of age or younger: clinical presentation, radiographic findings, management, and outcomes with a minimum 12-month follow-up.

Science.gov (United States)

Gressot, Loyola V; Vadivelu, Sudhakar; Hwang, Steven W; Fulkerson, Daniel H; Luerssen, Thomas G; Jea, Andrew

2014-02-01

Cervical spondylolysis is a rare condition that results from a pars interarticularis defect. The C-6 level is the most frequently involved site in the cervical spine. Its clinical presentations range from incidental radiographic findings to neck pain and, rarely, neurological deficits. Although 150 patients with subaxial cervical spondylolysis have been reported, a mere 24 adult and pediatric patients with C-2 spondylolysis have been described. The long-term outcomes of very young children with bilateral C-2 spondylolysis are of great interest, yet only a few longitudinal studies exist. The authors retrospectively reviewed 5 cases of bilateral C-2 spondylolysis at Texas Children's Hospital and Riley Children's Hospital; these were combined with 5 other cases in the literature, yielding a total of 10 patients. Data regarding the patients' age, sex, C2-3 angulation and displacement, associated spine anomalies, neurological deficits, treatment, and most recent follow-up were recorded. The patients' ages ranged from 3 to 36 months (mean 12.9 months). There were 6 boys and 4 girls. The C2-3 angulation, displacement, and width of pars defect were measured when available. The mean C2-3 angulation was 9.5° (range 1-34°), the mean C2-3 displacement was 4.78 mm (range 1.1-10.8 mm), and the mean width of the pars defect was 4.16 mm (range 0.9-7 mm). One patient developed myelopathy and spinal cord injury. All 10 of the patients were treated initially with conservative therapy: 3 with close observation alone, 1 with a rigid cervical collar, 4 with a Minerva jacket, 1 with a sternal-occipital-mandibular immobilizer, and 1 with a halo vest. Three patients ultimately underwent surgery for internal fixation due to progressive instability or development of neurological symptoms. All patients were neurologically intact at the last follow-up (mean 44.3 months, range 14-120 months). Based on the literature and the authors' own experience, they conclude that most very young children

Emission of particles in the {sup 12} C + {sup 12} C fusion; Emision de particulas en la fusion de {sup 12} C + {sup 12} C

Energy Technology Data Exchange (ETDEWEB)

Martinez Q, E.; Aguilera, E.F.; Rosales, P. [Instituto Nacional de Investigaciones Nucleares, A.P. 18-1027, 11801 Mexico D.F. (Mexico)

2002-07-01

A fusion process analysis of the {sup 12} C + {sup 12} C reaction is done, using the LILITA program. The analysis consisted mainly in varying the value of the Levels density parameter, determining on this way the value of such parameter which reproduces better the contribution of the different channels of fusion-evaporation of particles for this system at different energies. Moreover a comparison with measures done in the Instituto Nacional de Investigaciones Nucleares is realized. (Author)

properties of Cr(C,N) hard coatings deposited in Ar-C2H2-N2 plasma

International Nuclear Information System (INIS)

Macek, M.; Cekada, M.; Kek, D.; Panjan, P.

2002-01-01

Mechanical properties, microstructure and the average chemical composition of Cr(C,N) hard coatings deposited in Ar-C 2 H 2 -N 2 plasma strongly depends on the partial pressure of the reactive gases (N 2 , C 2 H 2 ) and on the type of the deposition equipment. In this study we report on the properties of Cr(C,N) hard coatings deposited by means of the triode ion plating in the BAI 730 apparatus and those prepared by sputter deposition in Balzers Sputron in the pressure range from 0.12 Pa (pure Ar) up to 0.35 Pa with different ratios (0-100%) between C 2 H 2 and N 2 . At first mechanical properties (microhardness and adhesion) of coatings were analyzed on the common way. Internal stress was measured by the radius of substrate curvature. Chemical composition of coatings was analyzed by means of AES while the Raman and XPS spectroscopy was used to determined the nature of carbon bonding in the Cr(C,N) films. Microstructure was determined by XRD as well as by means of TEM and TED. Chemical state of various elements in the coating has been studied by XPS. The ratio of the carbide bond (C-Cr) against the C-C and C-H bonds was calculated. The existence of the graphite phase in some Cr(C,N) coatings was confirmed by Raman spectroscopy. (Authors)

C-Cbl reverses HER2-mediated tamoxifen resistance in human breast cancer cells.

Science.gov (United States)

Li, Wei; Xu, Ling; Che, Xiaofang; Li, Haizhou; Zhang, Ye; Song, Na; Wen, Ti; Hou, Kezuo; Yang, Yi; Zhou, Lu; Xin, Xing; Xu, Lu; Zeng, Xue; Shi, Sha; Liu, Yunpeng; Qu, Xiujuan; Teng, Yuee

2018-05-02

Tamoxifen is a frontline therapy for estrogen receptor (ER)-positive breast cancer in premenopausal women. However, many patients develop resistance to tamoxifen, and the mechanism underlying tamoxifen resistance is not well understood. Here we examined whether ER-c-Src-HER2 complex formation is involved in tamoxifen resistance. MTT and colony formation assays were used to measure cell viability and proliferation. Western blot was used to detect protein expression and protein complex formations were detected by immunoprecipitation and immunofluorescence. SiRNA was used to examine the function of HER2 in of BT474 cells. An in vivo xenograft animal model was established to examine the role of c-Cbl in tumor growth. MTT and colony formation assay showed that BT474 cells are resistant to tamoxifen and T47D cells are sensitive to tamoxifen. Immunoprecipitation experiments revealed ER-c-Src-HER2 complex formation in BT474 cells but not in T47D cells. However, ER-c-Src-HER2 complex formation was detected after overexpressing HER2 in T47D cells and these cells were more resistant to tamoxifen. HER2 knockdown by siRNA in BT474 cells reduced ER-c-Src-HER2 complex formation and reversed tamoxifen resistance. ER-c-Src-HER2 complex formation was also disrupted and tamoxifen resistance was reversed in BT474 cells by the c-Src inhibitor PP2 and HER2 antibody trastuzumab. Nystatin, a lipid raft inhibitor, reduced ER-c-Src-HER2 complex formation and partially reversed tamoxifen resistance. ER-c-Src-HER2 complex formation was disrupted by overexpression of c-Cbl but not by the c-Cbl ubiquitin ligase mutant. In addition, c-Cbl could reverse tamoxifen resistance in BT474 cells, but the ubiquitin ligase mutant had no effect. The effect of c-Cbl was validated in BT474 tumor-bearing nude mice in vivo. Immunofluorescence also revealed ER-c-Src-HER2 complex formation was reduced in tumor tissues of nude mice with c-Cbl overexpression. Our results suggested that c-Cbl can reverse tamoxifen

c-Jun induces apoptosis of starved BM2 monoblasts by activating cyclin A-CDK2

International Nuclear Information System (INIS)

Vanhara, Petr; Bryja, Vitezslav; Horvath, Viktor; Kozubik, Alois; Hampl, Ales; Smarda, Jan

2007-01-01

c-Jun is one of the major components of the activating protein-1 (AP-1), the transcription factor that participates in regulation of proliferation, differentiation, and apoptosis. In this study, we explored functional interactions of the c-Jun protein with several regulators of the G1/S transition in serum-deprived v-myb-transformed chicken monoblasts BM2. We show that the c-Jun protein induces expression of cyclin A, thus up-regulating activity of cyclin A-associated cyclin-dependent kinase 2 (CDK2), and causing massive programmed cell death of starved BM2cJUN cells. Specific inhibition of CDK2 suppresses frequency of apoptosis of BM2cJUN cells. We conclude that up-regulation of cyclin A expression and CDK2 activity can represent important link between the c-Jun protein, cell cycle machinery, and programmed cell death pathway in leukemic cells

Neisseria meningitidis differentially controls host cell motility through PilC1 and PilC2 components of type IV Pili.

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Philippe C Morand

Full Text Available Neisseria meningitidis is a strictly human pathogen that has two facets since asymptomatic carriage can unpredictably turn into fulminant forms of infection. Meningococcal pathogenesis relies on the ability of the bacteria to break host epithelial or endothelial cellular barriers. Highly restrictive, yet poorly understood, mechanisms allow meningococcal adhesion to cells of only human origin. Adhesion of encapsulated and virulent meningococci to human cells relies on the expression of bacterial type four pili (T4P that trigger intense host cell signalling. Among the components of the meningococcal T4P, the concomitantly expressed PilC1 and PilC2 proteins regulate pili exposure at the bacterial surface, and until now, PilC1 was believed to be specifically responsible for T4P-mediated meningococcal adhesion to human cells. Contrary to previous reports, we show that, like PilC1, the meningococcal PilC2 component is capable of mediating adhesion to human ME180 epithelial cells, with cortical plaque formation and F-actin condensation. However, PilC1 and PilC2 promote different effects on infected cells. Cellular tracking analysis revealed that PilC1-expressing meningococci caused a severe reduction in the motility of infected cells, which was not the case when cells were infected with PilC2-expressing strains. The amount of both total and phosphorylated forms of EGFR was dramatically reduced in cells upon PilC1-mediated infection. In contrast, PilC2-mediated infection did not notably affect the EGFR pathway, and these specificities were shared among unrelated meningococcal strains. These results suggest that meningococci have evolved a highly discriminative tool for differential adhesion in specific microenvironments where different cell types are present. Moreover, the fine-tuning of cellular control through the combined action of two concomitantly expressed, but distinctly regulated, T4P-associated variants of the same molecule (i.e. PilC1 and PilC

Molecular dynamic simulation of the self-assembly of DAP12-NKG2C activating immunoreceptor complex.

Directory of Open Access Journals (Sweden)

Peng Wei

Full Text Available The DAP12-NKG2C activating immunoreceptor complex is one of the multisubunit transmembrane protein complexes in which ligand-binding receptor chains assemble with dimeric signal-transducing modules through non-covalent associations in their transmembrane (TM domains. In this work, both coarse grained and atomistic molecular dynamic simulation methods were applied to investigate the self-assembly dynamics of the transmembrane domains of the DAP12-NKG2C activating immunoreceptor complex. Through simulating the dynamics of DAP12-NKG2C TM heterotrimer and point mutations, we demonstrated that a five-polar-residue motif including: 2 Asps and 2 Thrs in DAP12 dimer, as well as 1 Lys in NKG2C TM plays an important role in the assembly structure of the DAP12-NKG2C TM heterotrimer. Furthermore, we provided clear evidences to exclude the possibility that another NKG2C could stably associate with the DAP12-NKG2C heterotrimer. Based on the simulation results, we proposed a revised model for the self-assembly of DAP12-NKG2C activating immunoreceptor complex, along with a plausible explanation for the association of only one NKG2C with a DAP12 dimer.

In vitro induction of O6-methylguanine-DNA methyltransferase in C3H/10T1/2 cells by X-rays is inhibited by nitrogen

International Nuclear Information System (INIS)

Hofe, E. von; Kennedy, A.R.

1988-01-01

Ionizing radiation is one of the most potent inducers of O 6 -methylguanine-DNA methyltransferase (MT) in rat liver in vivo. In this study we show that MT is readily induced in C3H/10T1/2 cells in culture, which provides a system more amenable to determining the molecular events involved in the induction of this repair enzyme. Maximal induction was observed in logarithmically growing cells 48 h after a dose of 200 rad, similar to the optimal induction time seen in rat liver in vivo. The absolute level of MT observed in C3H/10T1/2 cells which had been at confluence for 24 h was less than in cells in log growth but was still inducible by X-rays, exhibiting an ∼ 2-fold increase over unirradiated cells similar to MT induction in logarithmically growing cells. Irradiating cells under anaerobic conditions abolished MT induction by 100 rad. Cells irradiated with 200 rad under anaerobic conditions exhibited ∼ 70% inhibition of induction compared with aerobically irradiated cells. The possibility that MT may be partially inactivated by interaction with radicals produced by ionizing radiation is discussed. (author)

Fisetin induces G2/M phase cell cycle arrest by inactivating cdc25C-cdc2 via ATM-Chk1/2 activation in human endometrial cancer cells

Directory of Open Access Journals (Sweden)

Zhan-Ying Wang

2015-06-01

Full Text Available Endometrial cancer is one of the most prevalent gynaecological malignancies where, currently available therapeutic options remain limited. Recently phytochemicals are exploited for their efficiency in cancer therapy. The present study investigates the anti-proliferative effect of fisetin, a flavonoid on human endometrial cancer cells (KLE and Hec1 A. Fisetin (20-100 µM effectively reduced the viability of Hec1 A and KLE cells and potentially altered the cell population at G2/M stage. Expression levels of the cell cycle proteins (cyclin B1, p-Cdc2, p-Cdc25C, p-Chk1, Chk2, p-ATM, cyclin B1, H2AX, p21 and p27 were analyzed. Fisetin suppressed cyclin B1 expression and caused inactiva-tion of Cdc25C and Cdc2 by increasing their phosphorylation levels and further activated ATM, Chk1 and Chk2. Increased levels of p21 and p27 were observed as well. These results suggest that fisetin induced G2/M cell cycle arrest via inactivating Cdc25c and Cdc2 through activation of ATM, Chk1 and Chk2.

C3 rho-inhibitor for targeted pharmacological manipulation of osteoclast-like cells.

Directory of Open Access Journals (Sweden)

Andrea Tautzenberger

Full Text Available The C3 toxins from Clostridium botulinum (C3bot and Clostridium limosum (C3lim as well as C3-derived fusion proteins are selectively taken up into the cytosol of monocytes/macrophages where the C3-catalyzed ADP-ribosylation of Rho results in inhibition of Rho-signalling and characteristic morphological changes. Since the fusion toxin C2IN-C3lim was efficiently taken up into and inhibited proliferation of murine macrophage-like RAW 264.7 cells, its effects on RAW 264.7-derived osteoclasts were investigated. C2IN-C3lim was taken up into differentiated osteoclasts and decreased their resorption activity. In undifferentiated RAW 264.7 cells, C2IN-C3lim-treatment significantly decreased their differentiation into osteoclasts as determined by counting the multi-nucleated, TRAP-positive cells. This inhibitory effect was concentration- and time-dependent and most efficient when C2IN-C3lim was applied in the early stage of osteoclast-formation. A single-dose application of C2IN-C3lim at day 0 and its subsequent removal at day 1 reduced the number of osteoclasts in a comparable manner while C2IN-C3lim-application at later time points did not reduce the number of osteoclasts to a comparable degree. Control experiments with an enzymatically inactive C3 protein revealed that the ADP-ribosylation of Rho was essential for the observed effects. In conclusion, the results indicate that Rho-activity is crucial during the early phase of osteoclast-differentiation. Other bone cell types such as pre-osteoblastic cells were not affected by C2IN-C3lim. Due to their cell-type selective and specific mode of action, C3 proteins and C3-fusions might be valuable tools for targeted pharmacological manipulation of osteoclast formation and activity, which could lead to development of novel therapeutic strategies against osteoclast-associated diseases.

Remarks on surfaces with $c_1^2 =2\\chi -1$ having non-trivial 2-torsion

OpenAIRE

MURAKAMI, Masaaki

2013-01-01

We shall show that any complex minimal surface of general type with $c_1^2 = 2\\chi -1$ having non-trivial 2-torsion divisors, where $c_1^2$ and $\\chi$ are the first Chern number of a surface and the Euler characteristic of the structure sheaf respectively, has the Euler characteristic $\\chi$ not exceeding 4. Moreover, we shall give a complete description for the surfaces of the case $\\chi =4$, and prove that the coarse moduli space for surfaces of this case is a unirational variety of dimensi...

Vertebral artery variations at the C1-2 level diagnosed by magnetic resonance angiography

Energy Technology Data Exchange (ETDEWEB)

Uchino, Akira; Saito, Naoko; Watadani, Takeyuki; Okada, Yoshitaka; Kozawa, Eito; Nishi, Naoko; Mizukoshi, Waka; Inoue, Kaiji; Nakajima, Reiko; Takahashi, Masahiro [Saitama Medical University International Medical Center, Department of Diagnostic Radiology, Hidaka, Saitama (Japan)

2012-01-15

The craniovertebral junction is clinically important. The vertebral artery (VA) in its several variations runs within this area. We report the prevalence of these VA variations on magnetic resonance angiography (MRA). We retrospectively reviewed MRA images, obtained using two 1.5-T imagers, of 2,739 patients, and paid special attention to the course and branching of the VA at the level of the C1-2 vertebral bodies. There were three types of VA variation at the C1-2 level: (1) persistent first intersegmental artery (FIA), (2) VA fenestration, and (3) posterior inferior cerebellar artery (PICA) originating from the C1/2 level. The overall prevalence of these three variations was 5.0%. There was no laterality in frequency, but we found female predominance (P < 0.05). We most frequently observed the persistent FIA (3.2%), which was sometimes bilateral. We found VA fenestration (0.9%) and PICA of C1/2 origin (1.1%) with almost equal frequency. Two PICAs of C1/2 origin had no normal VA branch. We frequently observed VA variations at the C1-2 level and with female predominance. The persistent FIA was most prevalent and sometimes seen bilaterally. Preoperative identification of these variations in VA is necessary to avoid complications during surgery at the craniovertebral junction. (orig.)

Altered sensitivity of system A amino acid transport to ouabain in normal and transformed C3H-10T1/2 cells during the cell cycle

International Nuclear Information System (INIS)

Leister, K.J.; Schenerman, M.A.; Racker, E.

1989-01-01

Quiescent C3H-10T1/2 mouse fibroblasts that have not undergone any type of stress have a relatively low rate of 2-aminoisobutyrate (Aib) uptake by means of system A, which is primarily energized by the transmembrane Na + chemical gradient potential. System A activity in these cells is not sensitive to ouabain or proton ionophores. In contrast, methylcholanthrene-transformed and cofluent C3H-10T1/2 cells treated with ouabain utilize the membrane potential generated by the Na + , K + -ATPase pump to drive Aib transport by means of system A as shown by the sensitivity of transport activity to ouabain and proton ionophores. Since glucose is present during the assay, the proton ionophores do not affect the availability of ATP, as indicated by the undiminished uptake of 86 Rb + by the Na + , K + -ATPase pump. As cells progress through the G 1 phase of the cell cycle, they show an increased system A activity prior to entry into the S phase, which is also dependent on the electrogenicity of the Na + , K + -ATPase pump. There appears to be in all these cases a qualitative shift in the bioenergetic mechanism for the uptake of Aib as well as a marked quantitative increase in Aib uptake. The high activity after ouabain treatment was sustained in the transformed cells after removal of the ouabain, whereas in the confluent 10T1/2 cells the rate of uptake decayed rapidly, suggesting a difference in the mode of regulation. The authors conclude that transformed cells and normal cells in late G 1 or under stress make use of the membrane potential generated by the Na + , K + -ATPase pump to drive amino acid uptake by means of system A

Synergistic inhibition of the APC/C by the removal of APC15 in HCT116 cells lacking UBE2C.

Science.gov (United States)

Garvanska, Dimitriya H; Larsen, Marie Sofie Yoo; Nilsson, Jakob

2016-10-15

The spindle assembly checkpoint (SAC) inhibits the anaphase-promoting complex/cyclosome (APC/C) in response to unattached kinetochores by generating a diffusible inhibitor termed the mitotic checkpoint complex (MCC). At metaphase, rapid activation of the APC/C requires removal of the MCC, a process that has been shown to depend on the APC/C E2 enzymes, UBE2C and UBE2S. Here we investigate the in vivo role of the APC/C E2 enzymes in SAC silencing using CRISPR/Cas9 genetically engineered HCT116 UBE2C or UBE2S null cell lines. Using live cell assays, we show that UBE2C and UBE2S make a minor contribution to SAC silencing in HCT116 cells. Strikingly, in cells specifically lacking UBE2C, we observe a strong synergistic inhibition of mitotic progression when we stabilize the MCC on the APC/C by depleting APC15, potentially reflecting increased competition between the MCC and the remaining initiating E2 enzyme UBE2D. In conclusion, we provide in vivo insight into the APC/C E2 module and its interplay with SAC silencing components. © 2016. Published by The Company of Biologists Ltd.

Dafachronic acid inhibits C. elegans germ cell proliferation in a DAF-12-dependent manner.

Science.gov (United States)

Mukherjee, Madhumati; Chaudhari, Snehal N; Balachandran, Riju S; Vagasi, Alexandra S; Kipreos, Edward T

2017-12-15

Dafachronic acid (DA) is a bile acid-like steroid hormone that regulates dauer formation, heterochrony, and lifespan in C. elegans. Here, we describe that DA is an inhibitor of C. elegans germ stem cell proliferation in adult hermaphrodites. Using a C. elegans germ cell primary culture system, we show that DA inhibits the proliferation of germ cells in vitro. Exogenous DA reduces the frequency of large tumors in adult tumorous germline mutants and decreases the proliferation of wild-type germ stem cells in adult hermaphrodites. In contrast, DA has no appreciable effect on the proliferation of larval-stage germ cells in wild type. The inhibition of adult germ cell proliferation by DA requires its canonical receptor DAF-12. Blocking DA production by inactivating the cytochrome P450 DAF-9 increases germ cell proliferation in wild-type adult hermaphrodites and the frequency of large tumors in germline tumorous mutants, suggesting that DA inhibits the rate of germ cell proliferation under normal growth conditions. Copyright © 2017 Elsevier Inc. All rights reserved.

1-Nitropyrene (1-NP) induces apoptosis and apparently a non-apoptotic programmed cell death (paraptosis) in Hepa1c1c7 cells

International Nuclear Information System (INIS)

Asare, Nana; Landvik, Nina E.; Lagadic-Gossmann, Dominique; Rissel, Mary; Tekpli, Xavier; Ask, Kjetil; Lag, Marit; Holme, Jorn A.

2008-01-01

Mechanistic studies of nitro-PAHs (polycyclic aromatic hydrocarbons) of interest might help elucidate which chemical characteristics are most important in eliciting toxic effects. 1-Nitropyrene (1-NP) is the predominant nitrated PAH emitted in diesel exhaust. 1-NP-exposed Hepa1c1c7 cells exhibited marked changes in cellular morphology, decreased proliferation and different forms of cell death. A dramatic increase in cytoplasmic vacuolization was observed already after 6 h of exposure and the cells started to round up at 12 h. The rate of cell proliferation was markedly reduced at 24 h and apoptotic as well as propidium iodide (PI)-positive cells appeared. Electron microscopic examination revealed that the vacuolization was partly due to mitochondria swelling. The caspase inhibitor Z-VAD-FMK inhibited only the apoptotic cell death and Nec-1 (an inhibitor of necroptosis) exhibited no inhibitory effects on either cell death or vacuolization. In contrast, cycloheximide markedly reduced both the number of apoptotic and PI-positive cells as well as the cytoplasmic vacuolization, suggesting that 1-NP induced paraptotic cell death. All the MAPKs; ERK1/2, p38 and JNK, appear to be involved in the death process since marked activation was observed upon 1-NP exposure, and their inhibitors partly reduced the induced cell death. The ERK1/2 inhibitor PD 98057 completely blocked the induced vacuolization, whereas the other MAPKs inhibitors only had minor effects on this process. These findings suggest that 1-NP may cause apoptosis and paraptosis. In contrast, the corresponding amine (1-aminopyrene) elicited only minor apoptotic and necrotic cell death, and cells with characteristics typical of paraptosis were absent

Estradiol up-regulates L-type Ca2+ channels via membrane-bound estrogen receptor/phosphoinositide-3-kinase/Akt/cAMP response element-binding protein signaling pathway.

Science.gov (United States)

Yang, Xiaoyan; Mao, Xiaofang; Xu, Gao; Xing, Shasha; Chattopadhyay, Ansuman; Jin, Si; Salama, Guy

2018-05-01

In long QT syndrome type 2, women are more prone than men to the lethal arrhythmia torsades de pointes. We previously reported that 17β-estradiol (E2) up-regulates L-type Ca 2+ channels and current (I Ca,L ) (∼30%) in rabbit ventricular myocytes by a classic genomic mechanism mediated by estrogen receptor-α (ERα). In long QT syndrome type 2 (I Kr blockade or bradycardia), the higher Ca 2+ influx via I Ca,L causes Ca 2+ overload, spontaneous sarcoplasmic reticulum Ca 2+ release, and reactivation of I Ca,L that triggers early afterdepolarizations and torsades de pointes. The purpose of this study was to investigate the molecular mechanisms whereby E2 up-regulates I Ca,L , which are poorly understood. H9C2 and rat myocytes were incubated with E2 ± ER antagonist, or inhibitors of downstream transcription factors, for 24 hours, followed by western blots of Cav1.2α1C and voltage-clamp measurements of I Ca,L . Incubation of H9C2 cells with E2 (10-100 nM) increased I Ca,L density and Cav1.2α1C expression, which were suppressed by the ER antagonist ICI182,780 (1 μM). Enhanced I Ca,L and Cav1.2α1C expression by E2 was suppressed by inhibitors of phosphoinositide-3-kinase (Pi3K) (30 μM LY294002; P L via plasma membrane ER and by activating Pi3K, Akt, and CREB signaling. The promoter regions of the CACNA1C gene (human-rabbit-rat) contain adjacent/overlapping binding sites for p-CREB and ERα, which suggests a synergistic regulation by these pathways. Copyright © 2018 Heart Rhythm Society. Published by Elsevier Inc. All rights reserved.

Acute high-caffeine exposure increases autophagic flux and reduces protein synthesis in C2C12 skeletal myotubes.

Science.gov (United States)

Hughes, M A; Downs, R M; Webb, G W; Crocker, C L; Kinsey, S T; Baumgarner, Bradley L

2017-04-01

Caffeine is a highly catabolic dietary stimulant. High caffeine concentrations (1-10 mM) have previously been shown to inhibit protein synthesis and increase protein degradation in various mammalian cell lines. The purpose of this study was to examine the effect of short-term caffeine exposure on cell signaling pathways that regulate protein metabolism in mammalian skeletal muscle cells. Fully differentiated C2C12 skeletal myotubes either received vehicle (DMSO) or 5 mM caffeine for 6 h. Our analysis revealed that caffeine promoted a 40% increase in autolysosome formation and a 25% increase in autophagic flux. In contrast, caffeine treatment did not significantly increase the expression of the skeletal muscle specific ubiquitin ligases MAFbx and MuRF1 or 20S proteasome activity. Caffeine treatment significantly reduced mTORC1 signaling, total protein synthesis and myotube diameter in a CaMKKβ/AMPK-dependent manner. Further, caffeine promoted a CaMKII-dependent increase in myostatin mRNA expression that did not significantly contribute to the caffeine-dependent reduction in protein synthesis. Our results indicate that short-term caffeine exposure significantly reduced skeletal myotube diameter by increasing autophagic flux and promoting a CaMKKβ/AMPK-dependent reduction in protein synthesis.

Measurement of charged pions in {sup 12}C+{sup 12}C collisions at 1A GeV and 2A GeV with HADES

Energy Technology Data Exchange (ETDEWEB)

Agakishiev, G.; Destefanis, M.; Gilardi, C.; Kirschner, D.; Kuehn, W.; Lange, J.S.; Metag, V.; Novotny, R.; Pechenov, V.; Pechenova, O.; Perez Cavalcanti, T.; Spataro, S.; Spruck, B. [Justus Liebig Universitaet Giessen, II. Physikalisches Institut, Giessen (Germany); Agodi, C.; Coniglione, R.; Finocchiaro, P.; Maiolino, C.; Piattelli, P.; Sapienza, P. [Laboratori Nazionali del Sud, Istituto Nazionale di Fisica Nucleare, Catania (Italy); Balanda, A.; Kozuch, A.; Przygoda, W. [Jagiellonian University of Cracow, Smoluchowski Institute of Physics, Krakow (Poland); Panstwowa Wyzsza Szkola Zawodowa, Nowy Sacz (Poland); Bellia, G. [Laboratori Nazionali del Sud, Istituto Nazionale di Fisica Nucleare, Catania (Italy); Universita di Catania, Dipartimento di Fisica e Astronomia, Catania (Italy); Belver, D.; Cabanelas, P.; Duran, I.; Garzon, J.A.; Lamas-Valverde, J.; Marin, J. [University of Santiago de Compostela, Departamento de Fisica de Particulas, Santiago de Compostela (Spain); Belyaev, A.; Chernenko, S.; Fateev, O.; Ierusalimov, A.; Zanevsky, Y. [Joint Institute of Nuclear Research, Dubna (Russian Federation); Bielcik, J.; Braun-Munzinger, P.; Galatyuk, T.; Gonzalez-Diaz, D.; Heinz, T.; Holzmann, R.; Koenig, I.; Koenig, W.; Kolb, B.W.; Lang, S.; Muench, M.; Palka, M.; Pietraszko, J.; Rustamov, A.; Schroeder, C.; Schwab, E.; Simon, R.S.; Traxler, M.; Yurevich, S.; Zumbruch, P. [GSI Helmholtzzentrum fuer Schwerionenforschung, Darmstadt (Germany); Blanco, A.; Lopes, L.; Mangiarotti, A. [LIP-Laboratorio de Instrumentacao e Fisica Experimental de Particulas, Coimbra (Portugal); Bortolotti, A.; Michalska, B. [Sezione di Milano, Istituto Nazionale di Fisica Nucleare, Milano (Italy); Boyard, J.L.; Hennino, T.; Moriniere, E.; Ramstein, B.; Roy-Stephan, M.; Sudol, M. [CNRS/IN2P3 - Universite Paris Sud, Institut de Physique Nucleaire, Orsay Cedex (France); Christ, T.; Eberl, T.; Fabbietti, L.; Friese, J.; Gernhaeuser, R.; Jurkovic, M.; Kruecken, R. [and others

2009-04-15

We present the results of a study of charged-pion production in {sup 12}C+{sup 12}C collisions at incident beam energies of 1A GeV and 2A GeV using the HADES spectrometer at GSI. The main emphasis of the HADES program is on the dielectron signal from the early phase of the collision. Here, however, we discuss the data with respect to the emission of charged hadrons, specifically the production of {pi}{sup {+-}} mesons, which are related to neutral pions representing a dominant contribution to the dielectron yield. We have performed the first large-angular-range measurement of the distribution of {pi}{sup {+-}} mesons for the {sup 12}C+{sup 12}C collision system covering a fairly large rapidity interval. The pion yields, transverse-mass and angular distributions are compared with calculations done within a transport model, as well as with existing data from other experiments. The anisotropy of pion production is systematically analyzed. (orig.)

Early Decrease in Respiration and Uncoupling Event Independent of Cytochrome c Release in PC12 Cells Undergoing Apoptosis

Science.gov (United States)

Berghella, Libera; Ferraro, Elisabetta

2012-01-01

Cytochrome c is a key molecule in mitochondria-mediated apoptosis. It also plays a pivotal role in cell respiration. The switch between these two functions occurs at the moment of its release from mitochondria. This process is therefore extremely relevant for the fate of the cell. Since cytochrome c mediates respiration, we studied the changes in respiratory chain activity during the early stages of apoptosis in order to contribute to unravel the mechanisms of cytochrome c release. We found that, during staurosporine (STS)- induced apoptosis in PC12 cells, respiration is affected before the release of cytochrome c, as shown by a decrease in the endogenous uncoupled respiration and an uncoupling event, both occurring independently of cytochrome c release. The decline in the uncoupled respiration occurs also upon Bcl-2 overexpression (which inhibits cytochrome c release), while the uncoupling event is inhibited by Bcl-2. We also observed that the first stage of nuclear condensation during STS-induced apoptosis does not depend on the release of cytochrome c into the cytosol and is a reversibile event. These findings may contribute to understand the mechanisms affecting mitochondria during the early stages of apoptosis and priming them for the release of apoptogenic factors. PMID:22666257

TERAHERTZ SPECTROSCOPY AND GLOBAL ANALYSIS OF THE BENDING VIBRATIONS OF ACETYLENE 12C2D2

International Nuclear Information System (INIS)

Yu Shanshan; Drouin, Brian J.; Pearson, John C.; Pickett, Herbert M.; Lattanzi, Valerio; Walters, Adam

2009-01-01

Two hundred and fifty-one 12 C 2 D 2 transitions have been measured in the 0.2-1.6 THz region of its ν 5 -ν 4 difference band and 202 of them were observed for the first time. The accuracy of these measurements is estimated to be ranging from 50 kHz to 100 kHz. The 12 C 2 D 2 molecules were generated under room temperature by passing 120-150 mTorr D 2 O vapor through calcium carbide (CaC 2 ) powder. A multistate analysis was carried out for the bending vibrational modes ν 4 and ν 5 of 12 C 2 D 2 , which includes the lines observed in this work and prior microwave, far-infrared and infrared data on the pure bending levels. Significantly improved molecular parameters were obtained for 12 C 2 D 2 by adding the new measurements to the old data set, which had only 10 lines with microwave measurement precision. New frequency and intensity predictions have been made based on the obtained molecular parameters. The more precise measurements and new predictions reported here will support the analyses of astronomical observations by the future high-resolution spectroscopy telescopes such as Herschel, SOFIA, and ALMA, which will work in the terahertz spectral region.