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Sample records for multicellular spheroids exposed

  1. Unsuccessful mitosis in multicellular tumour spheroids.

    Science.gov (United States)

    Molla, Annie; Couvet, Morgane; Coll, Jean-Luc

    2017-04-25

    Multicellular spheroids are very attractive models in oncology because they mimic the 3D organization of the tumour cells with their microenvironment. We show here using 3 different cell types (mammary TSA/pc, embryonic kidney Hek293 and cervical cancer HeLa), that when the cells are growing as spheroids the frequency of binucleated cells is augmented as occurs in some human tumours.We therefore describe mitosis in multicellular spheroids by following mitotic markers and by time-lapse experiments. Chromosomes alignment appears to be correct on the metaphasic plate and the passenger complex is well localized on centromere. Moreover aurora kinases are fully active and histone H3 is phosphorylated on Ser 10. Consequently, the mitotic spindle checkpoint is satisfied and, anaphase proceeds as illustrated by the transfer of survivin on the spindle and by the segregation of the two lots of chromosomes. However, the segregation plane is not well defined and oscillations of the dividing cells are observed. Finally, cytokinesis fails and the absence of separation of the two daughter cells gives rise to binucleated cells.Division orientation is specified during interphase and persists throughout mitosis. Our data indicate that the cancer cells, in multicellular spheroids, lose their ability to regulate their orientation, a feature commonly encountered in tumours.Moreover, multicellular spheroid expansion is still sensitive to mitotic drugs as pactlitaxel and aurora kinase inhibitors. The spheroids thus represent a highly relevant model for studying drug efficiency in tumours.

  2. Mass Spectrometry Analyses of Multicellular Tumor Spheroids.

    Science.gov (United States)

    Acland, Mitchell; Mittal, Parul; Lokman, Noor A; Klingler-Hoffmann, Manuela; Oehler, Martin K; Hoffmann, Peter

    2018-05-01

    Multicellular tumor spheroids (MCTS) are a powerful biological in vitro model, which closely mimics the 3D structure of primary avascularized tumors. Mass spectrometry (MS) has established itself as a powerful analytical tool, not only to better understand and describe the complex structure of MCTS, but also to monitor their response to cancer therapeutics. The first part of this review focuses on traditional mass spectrometry approaches with an emphasis on elucidating the molecular characteristics of these structures. Then the mass spectrometry imaging (MSI) approaches used to obtain spatially defined information from MCTS is described. Finally the analysis of primary spheroids, such as those present in ovarian cancer, and the great potential that mass spectrometry analysis of these structures has for improved understanding of cancer progression and for personalized in vitro therapeutic testing is discussed. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  3. Multicellular spheroids as an in vitro tumor model

    International Nuclear Information System (INIS)

    Kozubek, S.; Erzgraber, G.

    1982-01-01

    Experiments with fractionated irradiation of multicellular spheroids were performed. Our data as well as the data of other works have been evaluated by means of simple mathematical formulae on the basis of several hypothesis. The spheroids are shown to exhibit similar behaviour as in vivo carcinomas. They offer the possibility of investigation of quantitative correlations for practical purposes

  4. Differential thermo-resistance of multicellular tumor spheroids

    International Nuclear Information System (INIS)

    Khoei, S.; Goliaei, B.; Neshasteh-Rize, A.

    2004-01-01

    Many cell lines, when cultured under proper conditions, can form three dimensional structures called multicellular spheroids. These spheroids resemble in vivo tumor models in several aspects. Therefore, studying growth characteristics and behavior of spheroids is beneficial in understanding the behavior of tumors under various experimental conditions. In this work, we have studied the growth properties, along with the thermal characteristics of spheroids of Du 145 human prostate carcinoma cell lines and compared the results to monolayer cultures of these cells. For this purpose, The Du 145 cells were cultured either as monolayer or spheroids. At various times after initiation of cultures, the growth properties of spheroids as a function of seeding cell number was determined. To evaluate the thermal characteristics of spheroids, they were heated at various stages of growth at 43 d ig c for various periods. The thermal response was judged by the survival fraction of colony forming cells in spheroids or monolayer culture following heat treatment. The results showed spheroids were more resistant to heat than monolayer cultures at all stages of development. However, the extent of this thermal resistant was dependent on the age, and consequently, the size of the spheroid. The result suggests that the differential thermal resistance of the spheroid cultures develop gradually during the growth of spheroid cultures of Du 145 cell line

  5. Cryopreservation of organotypic multicellular spheroids from human gliomas

    NARCIS (Netherlands)

    Kaaijk, P.; van den Berg, F.; van Amstel, P.; Troost, D.

    1996-01-01

    Fresh human glioma tissue can be cultured on agarose to form organotypic multicellular spheroids (OMS). The major advantage of OMS is the preservation of the cellular heterogeneity and the tumour architecture, which is lost in conventional monolayer cultures. The present study was undertaken to

  6. Heavy-ion radiobiology of multicellular tumor spheroids

    International Nuclear Information System (INIS)

    Rodriguez, A.; Alpen, E.L.

    1980-01-01

    Experiments reported here were conducted with carbon ions, neon ions, and argon ions using rat brain gliosarcoma (9L) and Chinese hamster lung V79 cells grown as multicellular spheroids. Our studies were designed to evaluate high-LET radiation survival characteristics of cells grown in this relatively organized tissue-like environment. Our primary objectives were to determine the RBE values in plateau and spread Bragg peak regions of the carbon, neon, and argon beams, and evaluate with high and low LET radiation, the role of spheroid architecture in postirradiation survival of cells grown in this format

  7. Response of the MG-63 human osteosarcoma cell line grown as multicellular spheroids to neutron irradiation

    International Nuclear Information System (INIS)

    Kubota, Nobuo; Kakehi, Masae; Matsubara, Shou; Koike, Sachiko; Ando, Koichi.

    1993-01-01

    Multicellular tumor spheroids are composed of the mixed populations of cells with regard to cell proliferation, nutrition, oxygenation and radiosensitivity. Human osteogenic sarcoma is generally considered clinically radioresistant. However, the in vitro cell survival curves for human osteogenic sarcoma cell lines do not differ from those of other tumor cell lines. In this study, the responses of human osteogenic sarcoma cell line to gamma ray and neutrons were investigated by using spheroid system. The spheroids of the osteogenic sarcoma cell line are considered to be a good in vitro model of radioresistant tumors. The purpose of this study is to measure the response of the spheroids to fast neutron irradiation. MG-63 human osteogenic sarcoma cell line was used for this study. The cell line was cultured in alpha-MEM with supplement. Cell survival was estimated after the trypsinization of spheroids 24 hours after irradiation. The method of measuring spheroid cure is explained. The mean number of surviving cells per spheroid can be obtained from the mean clonogenic number and cell survival curve. The cell survival of MG-63 spheroids exposed to gamma ray and neutrons and the dose effect curves for spheroid cure after irradiation are shown. (K.I.)

  8. Method for the determination of oxygen consumption rates and diffusion coefficients in multicellular spheroids

    OpenAIRE

    Mueller-Klieser, W.

    1984-01-01

    A method has been developed for the quantitative evaluation of oxygen tension (PO2) distributions in multicellular spheroids measured with O2-sensitive microelectrodes. The experimental data showed that multicellular tumor spheroids in stirred growth media were characterized by a diffusion-depleted zone surrounding the spheroids. This zone was elicited by an unstirred layer of medium next to the spheroid leading to a continuous decrease in the PO2 values from the bulk medium towards the spher...

  9. Radiobiological studies of cells in multicellular spheroids using a sequential trypsinization technique

    International Nuclear Information System (INIS)

    Giesbrecht, J.L.; Wilson, W.R.; Hill, R.P.

    1981-01-01

    The radiation response of V79 Chinese hamster cells grown as multicellular spheroids has been investigated by determining survival curves for treatment under a variety of different oxygen concentrations. Spheroids were irradiated under fully oxygenated conditions in air-equilibrated medium at 37 0 C, in medium exposed to lower oxygen tension (5% O 2 ) for times varying from 1 hr to 3 days, or under anoxic conditions. For comparison with the spheroids, using identical treatment conditions, V79 cells were grown in suspension as a subconfluent monolayer attached to Sephadex (microcarrier) beads and irradiated under fully oxygenated or anoxic conditions. The radiation response of cells at different depths within the spheroid was investigated by using a sequential trypsinization technique developed to remove eight or nine shells of cells successively from the spheroid surface. When irradiation was given under fully oxygenated conditions the outer few cell layers were more sensitive than the inner cells, a finding which is not understood. As expected the inner cells in spheroids irradiated in air (at 37 0 C) or in 5% O 2 are more resistant than the outer cells. For an acute exposure to 5% O 2 (1 hr) in the inner cells displayed full radiobiological hypoxia; however, for chronic exposures to low oxygen this was not the case. These results with the sequential trypsinization procedure suggest that the radiation response of cells in spheroids is more complex than anticipted

  10. Multicellular tumor spheroid interactions with bone cells and bone

    International Nuclear Information System (INIS)

    Wezeman, F.H.; Guzzino, K.M.; Waxler, B.

    1985-01-01

    In vitro coculture techniques were used to study HSDM1C1 murine fibrosarcoma multicellular tumor spheroid (HSDM1C1-MTS) interactions with mouse calvarial bone cells having osteoblastic characteristics and mouse bone explants. HSDM1C1-MTS attached to confluent bone cell monolayers and their attachment rate was quantified. HSDM1C1-MTS interaction with bone cells was further demonstrated by the release of 3 H-deoxyuridine from prelabeled bone cells during coculture with multicellular tumor spheroids. HSDM1C1-MTS-induced cytotoxicity was mimicked by the addition of 10(-5) M prostaglandin E2 (PGE2) to 3 H-deoxyuridine-labeled bone cells. The effects of low (10(-9) M) and high (10(-5) M) concentrations of PGE2 on bone cell proliferation were also studied. Higher concentrations of PGE2 inhibited bone cell proliferation. HSDM1C1-MTS resorbed living explants in the presence of indomethacin, suggesting that other tumor cell products may also participate in bone resorption. HSDM1C1-MTS caused direct bone resorption as measured by the significantly elevated release of 45 Ca from prelabeled, devitalized calvaria. However, the growth of a confluent bone cell layer on devitalized, 45 Ca-prelabeled calvaria resulted in a significant reduction in the amount of 45 Ca released subsequent to the seeding of HSDM1C1-MTS onto the explants. Bone cells at the bone surface may act as a barrier against invasion and tumor cell-mediated bone resorption. Violation of this cellular barrier is achieved, in part, by tumor cell products

  11. Biomaterial Substrate-Mediated Multicellular Spheroid Formation and Their Applications in Tissue Engineering.

    Science.gov (United States)

    Tseng, Ting-Chen; Wong, Chui-Wei; Hsieh, Fu-Yu; Hsu, Shan-Hui

    2017-12-01

    Three-dimentional (3D) multicellular aggregates (spheroids), compared to the traditional 2D monolayer cultured cells, are physiologically more similar to the cells in vivo. So far there are various techniques to generate 3D spheroids. Spheroids obtained from different methods have already been applied to regenerative medicine or cancer research. Among the cell spheroids created by different methods, the substrate-derived spheroids and their forming mechanism are unique. This review focuses on the formation of biomaterial substrate-mediated multicellular spheroids and their applications in tissue engineering and tumor models. First, the authors will describe the special chitosan substrate-derived mesenchymal stem cell (MSC) spheroids and their greater regenerative capacities in various tissues. Second, the authors will describe tumor spheroids derived on chitosan and hyaluronan substrates, which serve as a simple in vitro platform to study 3D tumor models or to perform cancer drug screening. Finally, the authors will mention the self-assembly process for substrate-derived multiple cell spheroids (co-spheroids), which may recapitulate the heterotypic cell-cell interaction for co-cultured cells or crosstalk between different types of cells. These unique multicellular mono-spheroids or co-spheroids represent a category of 3D cell culture with advantages of biomimetic cell-cell interaction, better functionalities, and imaging possibilities. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  12. Synergistic interaction between cisplatin and gemcitabine in neuroblastoma cell lines and multicellular tumor spheroids

    NARCIS (Netherlands)

    Besançon, Odette G.; Tytgat, Godelieve A. M.; Meinsma, Rutger; Leen, René; Hoebink, Jerry; Kalayda, Ganna V.; Jaehde, Ulrich; Caron, Huib N.; van Kuilenburg, André B. P.

    2012-01-01

    The efficacy and mechanism of action of cisplatin and gemcitabine were investigated in a panel of neuroblastoma cell lines and multicellular tumor spheroids. In neuroblastoma spheroids, the combination of cisplatin and gemcitabine induced a complete cytostasis at clinical relevant concentrations. A

  13. Cell shedding from X-irradiated multicellular spheroids of human lung carcinomas

    International Nuclear Information System (INIS)

    Sakata, K.; Okada, S.; Suzuki, N.; Majima, H.

    1991-01-01

    We studied the effect of radiation on cell shedding from the surface of multicellular spheroids. Spheroids were produced from two human lung cell lines, one adenocarcinoma (LCT1) and the other small cell carcinoma (LCT2), by using a liquid overlay culture technique. The number of cells shed from both kinds of spheroids did not change significantly when they were irradiated. The number of clonogenic cells shed from both kinds of irradiated spheroids decreased sharply as the dose of irradiation increases. There were no significant differences in clonogenic cell shedding per spheroid between LCT1 and LCT2 spheroids. 400 μm spheroids were more radioresistant to inhibition of clonogenic cell shedding than 250 μm spheroids. Shed cells were more radiosensitive than speroid cells. In these experiments, we did not obtain any results indicating that radiation enchances metastasis. (orig.) [de

  14. Radiobiological investigations of multicellular spheroids as an in vitro tumor model. 4

    International Nuclear Information System (INIS)

    Kopp, J.

    1978-01-01

    Multicellular spheroids of various size were irradiated with a single dose of X-rays or fast neutrons, incubated after irradiation in roller tubes for long time, and observed light-microscopically. The observed effects were found independent of the radiation used and dependent on dose (considering the RBE) and size of spheroids. After irradiation the spheroid surface showed a loosening phenomenon and the spheroid structure changed depending on dose in three various types. Up to the 10% survival fraction the spheroids recovered within some days to the structure of non-irradiated spheroids by the growth of the surviving cells. After higher doses single surviving cells inside the dead spheroid material seemed to migrate from the inner hypoxic into the rim zone. They can start proliferation many days after irradiation leading to repopulation of irradiated spheroids. The volume changes of irradiated spheroids were also dose-dependent, but the measurement of spheroid volume seems to be of limited value for predicting the effectiveness of irradiation because the spheroid volume of irradiated spheroids is not proportional to the number of cells per spheroid. (author)

  15. Rapid formation of size-controllable multicellular spheroids via 3D acoustic tweezers

    OpenAIRE

    Chen, Kejie; Wu, Mengxi; Guo, Feng; Li, Peng; Chan, Chung-Yu; Mao, Zhangming; Li, Sixing; Ren, Liqiang; Zhang, Rui; Huang, Tony Jun

    2016-01-01

    The multicellular spheroid is an important 3D cell culture model for drug screening, tissue engineering, and fundamental biological research. Although several spheroid formation methods have been reported, the field still lacks high-throughput and simple fabrication methods to accelerate its adoption in drug development industry. Surface acoustic wave (SAW) based cell manipulation methods, which are known to be non-invasive, flexible, and high-throughput, have not been successfully developed ...

  16. Daunorubicin and doxorubicin but not BCNU have deleterious effects on organotypic multicellular spheroids of gliomas

    NARCIS (Netherlands)

    Kaaijk, P.; Troost, D.; de Boer, O. J.; van Amstel, P.; Bakker, P. J.; Leenstra, S.; Bosch, D. A.

    1996-01-01

    In the present study organotypic multicellular spheroids (OMS) were used to study the effects of chemotherapeutic agents on malignant gliomas. Compared with the frequently used cell line models, OMS have several advantages with respect to the preservation of the cellular heterogeneity and the

  17. Radiosensitivity of different human tumor cells lines grown as multicellular spheroids determined from growth curves and survival data

    International Nuclear Information System (INIS)

    Schwachoefer, J.H.C.; Crooijmans, R.P.; van Gasteren, J.J.; Hoogenhout, J.; Jerusalem, C.R.; Kal, H.B.; Theeuwes, A.G.

    1989-01-01

    Five human tumor cell lines were grown as multicellular tumor spheroids (MTS) to determine whether multicellular tumor spheroids derived from different types of tumors would show tumor-type dependent differences in response to single-dose irradiation, and whether these differences paralleled clinical behavior. Multicellular tumor spheroids of two neuroblastoma, one lung adenocarcinoma, one melanoma, and a squamous cell carcinoma of the oral tongue, were studied in terms of growth delay, calculated cell survival, and spheroid control dose50 (SCD50). Growth delay and cell survival analysis for the tumor cell lines showed sensitivities that correlated well with clinical behavior of the tumor types of origin. Similar to other studies on melanoma multicellular tumor spheroids our spheroid control dose50 results for the melanoma cell line deviated from the general pattern of sensitivity. This might be due to the location of surviving cells, which prohibits proliferation of surviving cells and hence growth of melanoma multicellular tumor spheroids. This study demonstrates that radiosensitivity of human tumor cell lines can be evaluated in terms of growth delay, calculated cell survival, and spheroid control dose50 when grown as multicellular tumor spheroids. The sensitivity established from these evaluations parallels clinical behavior, thus offering a unique tool for the in vitro analysis of human tumor radiosensitivity

  18. Oxygenation and response to irradiation of organotypic multicellular spheroids of human glioma.

    Science.gov (United States)

    Sminia, Peter; Acker, Helmut; Eikesdal, Hans Petter; Kaaijk, Patricia; Enger, Per øvind; Slotman, Ben; Bjerkvig, Rolf

    2003-01-01

    Investigation of the oxygenation status of organotypic multicellular spheroids (OMS) and their response to irradiation. Tumour specimens of glioblastoma multiforme patients (n = 16) were initiated as OMS. Following 20 Gy gamma-irradiation, the cell migratory capacity was evaluated. Spheroid oxygenation was determined by micro-electrode pO2 measurements and pimonidazole immunostaining. Spheroids prepared from established human glioma cell lines were used as a reference. Irradiation inhibited spheroid outgrowth by 12 to 88% relative to the non-irradiated controls. A large interpatient variation was noticed. Oxygen measurements revealed a gradual decrease in pO2 level from the periphery to the core of the spheroids, but the pO2 values remained within an oxygenated range. However, in the cell line spheroids an intermediate layer of hypoxia surrounding the central core was observed. Cell line spheroids with a hypoxic cell fraction and well-oxygenated OMS both show high resistance to irradiation, indicating that hypoxia may not be the biological factor determining the radioresistance of glioma spheroids in vitro.

  19. Rapid formation of size-controllable multicellular spheroids via 3D acoustic tweezers.

    Science.gov (United States)

    Chen, Kejie; Wu, Mengxi; Guo, Feng; Li, Peng; Chan, Chung Yu; Mao, Zhangming; Li, Sixing; Ren, Liqiang; Zhang, Rui; Huang, Tony Jun

    2016-07-05

    The multicellular spheroid is an important 3D cell culture model for drug screening, tissue engineering, and fundamental biological research. Although several spheroid formation methods have been reported, the field still lacks high-throughput and simple fabrication methods to accelerate its adoption in drug development industry. Surface acoustic wave (SAW) based cell manipulation methods, which are known to be non-invasive, flexible, and high-throughput, have not been successfully developed for fabricating 3D cell assemblies or spheroids, due to the limited understanding on SAW-based vertical levitation. In this work, we demonstrated the capability of fabricating multicellular spheroids in the 3D acoustic tweezers platform. Our method used drag force from microstreaming to levitate cells in the vertical direction, and used radiation force from Gor'kov potential to aggregate cells in the horizontal plane. After optimizing the device geometry and input power, we demonstrated the rapid and high-throughput nature of our method by continuously fabricating more than 150 size-controllable spheroids and transferring them to Petri dishes every 30 minutes. The spheroids fabricated by our 3D acoustic tweezers can be cultured for a week with good cell viability. We further demonstrated that spheroids fabricated by this method could be used for drug testing. Unlike the 2D monolayer model, HepG2 spheroids fabricated by the 3D acoustic tweezers manifested distinct drug resistance, which matched existing reports. The 3D acoustic tweezers based method can serve as a novel bio-manufacturing tool to fabricate complex 3D cell assembles for biological research, tissue engineering, and drug development.

  20. IN VITRO INVESTIGATION OF THE TRANSPLANTATION PROSPECTS OF MULTICELLULAR SPHEROID MICROAGGREGATES OF DONOR RETINAL PIGMENT EPITHELIUM

    Directory of Open Access Journals (Sweden)

    S. A. Borzenok

    2015-01-01

    Full Text Available Aim. To study in experiment the criteria for transplantability of multicellular spheroid microaggregates of retinal pigment epithelium (RPE, prepared by the method of 3D cell culture. Materials and Methods. 11 donor eyes (6 of adrenaline index «A», 5 of index «B» were used as a source of RPE cell cultures (group «A» – 6 cultures, group «B» – 5 cultures, of which over 2000 RPE spheroids were obtained by the method of three-dimensional cell culture. 1760 spheroids of them were selected for transplantability investigation (960 – group «A», 800 – group «B». Among the selected spheroids were equal numbers of spheroids of different morphology («smooth» and «rough» and of the initial cell seeding number (500, 1000, 5000, 25 000, 125 000 cells per hanging drop. We were taking out 12 spheroids of group «A» and 10 spheroids of group «B» of the 3D culture in terms of 7, 14, 21, 28 days of 3D culture to assess their viability. We were transferring the same number of spheroids in the same terms from 3D to 2D culture conditions to assess their adhesive properties. Viability of cells within spheroids was determined using the Trypan blue exclusion. The presence or absence of adhesion was determined by microscopic observation.Results. «Smooth» spheroids of 7 and 14 days of pretransplantation cultivation and derived from hanging drops containing 500 and 1000 cells showed the highest transplantability (cell viability varied from 0.83 ± 0.38 to 0.94 ± 0.24, a 100% adhesion. «Rough» spheroids were untransplantable in all variants, despite their partial preservation of viability (in comparison to “smooth” ones p < 0.05. 21 and 28 days of pretransplantation culturing and high cell seeding numbers signifi cantly lowered transplantability of obtained spheroids (p > 0.05 for low cell numbers, p < 0.05 for the high ones. Differences in adrenaline indexes A and B of donor eyes which were the primary sources of cellular

  1. Oxygen consumption rate and mitochondrial density in human melanoma monolayer cultures and multicellular spheroids.

    Science.gov (United States)

    Hystad, M E; Rofstad, E K

    1994-05-15

    Rate of oxygen consumption per cell has been shown in previous studies to decrease with increasing depth in the viable rim of multicellular spheroids initiated from rodent cells, human colon-carcinoma cells, and human glioma cells, due to progressive accumulation of quiescent cells during spheroid growth. The purpose of our work was to determine oxygen-consumption profiles in human melanoma spheroids. Monolayer cultures of 4 lines (BEX-c, COX-c, SAX-c, and WIX-c) and spheroid cultures of 2 lines (BEX-c and WIX-c) were subjected to investigation. Spheroids were initiated from monolayer cell cultures and grown in spinner flasks. Rate of oxygen consumption was measured with a Clarke-type electrode. Mitochondrial density was determined by stereological analysis of transmission electron micrographs. Thickness of viable rim and cell packing density were assessed by light microscopy of central spheroid sections. Cell-cycle distribution was determined by analysis of DNA histograms measured by flow cytometry. Cell volume was measured by an electronic particle counter. Rate of oxygen consumption per cell differed by a factor of approximately 1.8 between the 4 cell lines and was positively correlated to total volume of mitochondria per cell. Rate of oxygen consumption per cell and total volume of mitochondria per cell were equal for monolayer cell cultures, 600-microns spheroids and 1,200-microns spheroids of the same line. Mitochondrial density and location in the cell did not differ between cells at the spheroid surface, in the middle of the viable rim and adjacent to the central necrosis. Cell-cycle distribution, cell volume, and cell-packing density in the outer and inner halves of the viable rim were not significantly different. Consequently, the rate of oxygen consumption per cell in inner regions of the viable rim was probably equal to that at the spheroid surface, suggesting that oxygen diffusion distances may be shorter in some melanomas than in many other tumor

  2. Generation of Multicellular Tumor Spheroids with Microwell-Based Agarose Scaffolds for Drug Testing.

    Directory of Open Access Journals (Sweden)

    Xue Gong

    Full Text Available Three dimensional multicellular aggregate, also referred to as cell spheroid or microtissue, is an indispensable tool for in vitro evaluating antitumor activity and drug efficacy. Compared with classical cellular monolayer, multicellular tumor spheroid (MCTS offers a more rational platform to predict in vivo drug efficacy and toxicity. Nevertheless, traditional processing methods such as plastic dish culture with nonadhesive surfaces are regularly time-consuming, laborious and difficult to provide uniform-sized spheroids, thus causing poor reproducibility of experimental data and impeding high-throughput drug screening. In order to provide a robust and effective platform for in vitro drug evaluation, we present an agarose scaffold prepared with the template containing uniform-sized micro-wells in commercially available cell culture plates. The agarose scaffold allows for good adjustment of MCTS size and large-scale production of MCTS. Transparent agarose scaffold also allows for monitoring of spheroid formation under an optical microscopy. The formation of MCTS from MCF-7 cells was prepared using different-size-well templates and systematically investigated in terms of spheroid growth curve, circularity, and cell viability. The doxorubicin cytotoxicity against MCF-7 spheroid and MCF-7 monolayer cells was compared. The drug penetration behavior, cell cycle distribution, cell apoptosis, and gene expression were also evaluated in MCF-7 spheroid. The findings of this study indicate that, compared with cellular monolayer, MCTS provides a valuable platform for the assessment of therapeutic candidates in an in vivo-mimic microenvironment, and thus has great potential for use in drug discovery and tumor biology research.

  3. Characterization of variants isolated from BCNU-treated 9L multicellular spheroids

    International Nuclear Information System (INIS)

    Hoff, M.H.B.; Deen, D.F.

    1984-01-01

    Multicellular spheroids of the 9L rat brain tumor cell line were treated with a single high-dose to produce cells of varying sensitivity. Treatment of 350-450 μm diameter 9L spheroids with 45 μM BCNU for 1 hr at 37 0 C produced 5 log cell kill. Some of the treated spheroids were dissociated immediately after treatment; others were dissociated after 1 wk. From these populations, twenty clones were selected and passaged as monolayers. Each clone was assayed at passage 2 for BCNU-induced damage using colony forming efficiency and sister chromatid exchange. 60% of the clones were resistant to BCNU, 15% were unchanged, and 15% appeared hypersensitive as compared with the control, which were uncloned 9L cells. All of the hypersensitive clones originated from the spheroids that remained in suspension after treatment. Three clones were studied during subsequent passage in monolayer. Two resistant clones maintained their resistance to BCNU over -- 25 passages, while one hypersensitive clone appeared to become progressively more resistant during passage. Thus, as with monolayer and in vivo 9L cells, a single high-dose treatment with BCNU produces a spectrum of sensitivities to BCNU. Some of these phenotypes are stable over many passages and have been used to initiate 9L spheroids having varying sensitivity to BCNU. These spheroids will be used to investigate drug-radiation interactions

  4. Radiation cell survival and growth delay studies in multicellular spheroids of small-cell lung carcinoma

    International Nuclear Information System (INIS)

    Duchesne, G.M.; Peacock, J.H.

    1987-01-01

    The radiation sensitivity of two small-cell lung carcinoma cell lines growing as multicellular spheroids in static culture was determined using clonogenic cell survival and growth delay as endpoints. Growth delay determination suggested that clonogenic cell kill was less than was obtained by direct assay of cell survival. Recovery from potentially lethal damage was assayed in one line (HC12) but was not demonstrable, and clonogenic cell survival decreased with time in treated spheroids with diameters greater than 300 μm which contained a hypoxic cell population. Microscopic examination of the treated spheroids showed the emergence of an abnormal giant-cell population, and the progressive clonogenic cell loss that occurred after treatment was thought to be due to oxygen and nutrient deprivation of the remaining viable cells by this doomed cell population. Correction of the growth delay measurements for changes in cell size and clonogenic cell population allowed correlation of the growth delay and cell survival data. (author)

  5. A 3D printed microfluidic perfusion device for multicellular spheroid cultures.

    Science.gov (United States)

    Ong, Louis Jun Ye; Islam, Anik; DasGupta, Ramanuj; Iyer, Narayanan Gopalakkrishna; Leo, Hwa Liang; Toh, Yi-Chin

    2017-09-11

    The advent of 3D printing technologies promises to make microfluidic organ-on-chip technologies more accessible for the biological research community. To date, hydrogel-encapsulated cells have been successfully incorporated into 3D printed microfluidic devices. However, there is currently no 3D printed microfluidic device that can support multicellular spheroid culture, which facilitates extensive cell-cell contacts important for recapitulating many multicellular functional biological structures. Here, we report a first instance of fabricating a 3D printed microfluidic cell culture device capable of directly immobilizing and maintaining the viability and functionality of 3D multicellular spheroids. We evaluated the feasibility of two common 3D printing technologies i.e. stereolithography (SLA) and PolyJet printing, and found that SLA could prototype a device comprising of cell immobilizing micro-structures that were housed within a microfluidic network with higher fidelity. We have also implemented a pump-free perfusion system, relying on gravity-driven flow to perform medium perfusion in order to reduce the complexity and footprint of the device setup, thereby improving its adaptability into a standard biological laboratory. Finally, we demonstrated the biological performance of the 3D printed device by performing pump-free perfusion cultures of patient-derived parental and metastatic oral squamous cell carcinoma tumor and liver cell (HepG2) spheroids with good cell viability and functionality. This paper presents a proof-of-concept in simplifying and integrating the prototyping and operation of a microfluidic spheroid culture device, which will facilitate its applications in various drug efficacy, metabolism and toxicity studies.

  6. Vorinostat eliminates multicellular resistance of mesothelioma 3D spheroids via restoration of Noxa expression.

    Directory of Open Access Journals (Sweden)

    Dario Barbone

    Full Text Available When grown in 3D cultures as spheroids, mesothelioma cells acquire a multicellular resistance to apoptosis that resembles that of solid tumors. We have previously found that resistance to the proteasome inhibitor bortezomib in 3D can be explained by a lack of upregulation of Noxa, the pro-apoptotic BH3 sensitizer that acts via displacement of the Bak/Bax-activator BH3-only protein, Bim. We hypothesized that the histone deacetylase inhibitor vorinostat might reverse this block to Noxa upregulation in 3D. Indeed, we found that vorinostat effectively restored upregulation of Noxa protein and message and abolished multicellular resistance to bortezomib in the 3D spheroids. The ability of vorinostat to reverse resistance was ablated by knockdown of Noxa or Bim, confirming the essential role of the Noxa/Bim axis in the response to vorinostat. Addition of vorinostat similarly increased the apoptotic response to bortezomib in another 3D model, the tumor fragment spheroid, which is grown from human mesothelioma ex vivo. In addition to its benefit when used with bortezomib, vorinostat also enhanced the response to cisplatin plus pemetrexed, as shown in both 3D models. Our results using clinically relevant 3D models show that the manipulation of the core apoptotic repertoire may improve the chemosensitivity of mesothelioma. Whereas neither vorinostat nor bortezomib alone has been clinically effective in mesothelioma, vorinostat may undermine chemoresistance to bortezomib and to other therapies thereby providing a rationale for combinatorial strategies.

  7. Vorinostat Eliminates Multicellular Resistance of Mesothelioma 3D Spheroids via Restoration of Noxa Expression

    Science.gov (United States)

    Barbone, Dario; Cheung, Priscilla; Battula, Sailaja; Busacca, Sara; Gray, Steven G.; Longley, Daniel B.; Bueno, Raphael; Sugarbaker, David J.; Fennell, Dean A.; Broaddus, V. Courtney

    2012-01-01

    When grown in 3D cultures as spheroids, mesothelioma cells acquire a multicellular resistance to apoptosis that resembles that of solid tumors. We have previously found that resistance to the proteasome inhibitor bortezomib in 3D can be explained by a lack of upregulation of Noxa, the pro-apoptotic BH3 sensitizer that acts via displacement of the Bak/Bax-activator BH3-only protein, Bim. We hypothesized that the histone deacetylase inhibitor vorinostat might reverse this block to Noxa upregulation in 3D. Indeed, we found that vorinostat effectively restored upregulation of Noxa protein and message and abolished multicellular resistance to bortezomib in the 3D spheroids. The ability of vorinostat to reverse resistance was ablated by knockdown of Noxa or Bim, confirming the essential role of the Noxa/Bim axis in the response to vorinostat. Addition of vorinostat similarly increased the apoptotic response to bortezomib in another 3D model, the tumor fragment spheroid, which is grown from human mesothelioma ex vivo. In addition to its benefit when used with bortezomib, vorinostat also enhanced the response to cisplatin plus pemetrexed, as shown in both 3D models. Our results using clinically relevant 3D models show that the manipulation of the core apoptotic repertoire may improve the chemosensitivity of mesothelioma. Whereas neither vorinostat nor bortezomib alone has been clinically effective in mesothelioma, vorinostat may undermine chemoresistance to bortezomib and to other therapies thereby providing a rationale for combinatorial strategies. PMID:23300762

  8. Real-time viability and apoptosis kinetic detection method of 3D multicellular tumor spheroids using the Celigo Image Cytometer.

    Science.gov (United States)

    Kessel, Sarah; Cribbes, Scott; Bonasu, Surekha; Rice, William; Qiu, Jean; Chan, Leo Li-Ying

    2017-09-01

    The development of three-dimensional (3D) multicellular tumor spheroid models for cancer drug discovery research has increased in the recent years. The use of 3D tumor spheroid models may be more representative of the complex in vivo tumor microenvironments in comparison to two-dimensional (2D) assays. Currently, viability of 3D multicellular tumor spheroids has been commonly measured on standard plate-readers using metabolic reagents such as CellTiter-Glo® for end point analysis. Alternatively, high content image cytometers have been used to measure drug effects on spheroid size and viability. Previously, we have demonstrated a novel end point drug screening method for 3D multicellular tumor spheroids using the Celigo Image Cytometer. To better characterize the cancer drug effects, it is important to also measure the kinetic cytotoxic and apoptotic effects on 3D multicellular tumor spheroids. In this work, we demonstrate the use of PI and caspase 3/7 stains to measure viability and apoptosis for 3D multicellular tumor spheroids in real-time. The method was first validated by staining different types of tumor spheroids with PI and caspase 3/7 and monitoring the fluorescent intensities for 16 and 21 days. Next, PI-stained and nonstained control tumor spheroids were digested into single cell suspension to directly measure viability in a 2D assay to determine the potential toxicity of PI. Finally, extensive data analysis was performed on correlating the time-dependent PI and caspase 3/7 fluorescent intensities to the spheroid size and necrotic core formation to determine an optimal starting time point for cancer drug testing. The ability to measure real-time viability and apoptosis is highly important for developing a proper 3D model for screening tumor spheroids, which can allow researchers to determine time-dependent drug effects that usually are not captured by end point assays. This would improve the current tumor spheroid analysis method to potentially better

  9. Apoptosis, energy metabolism, and fraction of radiobiologically hypoxic cells: a study of human melanoma multicellular spheroids.

    Science.gov (United States)

    Rofstad, E K; Eide, K; Skøyum, R; Hystad, M E; Lyng, H

    1996-09-01

    The magnitude of the fraction of radiobiologically hypoxic cells in tumours is generally believed to reflect the efficiency of the vascular network. Theoretical studies have suggested that the hypoxic fraction might also be influenced by biological properties of the tumour cells. Quantitative experimental results of cell energy metabolism, hypoxia- induced apoptosis, and radiobiological hypoxia are reported here. Human melanoma multicellular spheroids (BEX-c and WIX-c) were used as tumour models to avoid confounding effects of the vascular network. Radiobiological studies showed that the fractions of hypoxic cells in 1000-microM spheroids were 32 +/- 12% (BEX-c) and 2.5 +/- 1.1% (WIX-c). The spheroid hypoxic volume fractions (28 +/- 6% (BEX-c) and 1.4 +/- 7% (WIX-c)), calculated from the rate of oxygen consumption per cell, the cell packing density, and the thickness of the viable rim, were similar to the fractions of radiobiologically hypoxic cells. Large differences between tumours in fraction of hypoxic cells are therefore not necessarily a result of differences in the efficiency of the vascular network. Studies of monolayer cell cultures, performed to identify the biological properties of the BEX-c and WIX-c cells leading to this large difference in fraction of hypoxic cells, gave the following results: (1) WIX-c showed lower cell surviving fractions after exposure to hypoxia than BEX-c, (2) WIX-c showed higher glucose uptake and lactate release rates than BEX-c both under aerobic and hypoxic conditions, and (3) hypoxia induced apoptosis in WIX-c but not in BEX-c. These observations suggested that the difference between BEX-c and WIX-c spheroids in fraction of hypoxic cells resulted partly from differences in cell energy metabolism and partly from a difference in capacity to retain viability under hypoxic stress. The induction of apoptosis by hypoxia was identified as a phenomenon which has an important influence on the magnitude of the fraction of

  10. Hypofractionated radiation induces a decrease in cell proliferation but no histological damage to organotypic multicellular spheroids of human glioblastomas

    NARCIS (Netherlands)

    Kaaijk, P.; Troost, D.; Sminia, P.; Hulshof, M. C.; van der Kracht, A. H.; Leenstra, S.; Bosch, D. A.

    1997-01-01

    The aim of this study was to examine the effect of radiation on glioblastoma, using an organotypic multicellular spheroid (OMS) model. Most glioblastoma cell lines are, in contrast to glioblastomas in vivo, relatively radiosensitive. This limits the value of using cell lines for studying the

  11. Cytolytic effects of autologous lymphokine-activated killer cells on organotypic multicellular spheroids of gliomas in vitro

    NARCIS (Netherlands)

    Kaaijk, P.; Troost, D.; Dast, P. K.; van den Berg, F.; Leenstra, S.; Bosch, D. A.

    1995-01-01

    Knowledge about lymphokine-activated killer (LAK) cell infiltration and LAK cell cytotoxicity is essential to improve the effectiveness of LAK cell therapy against gliomas. In the present study, organotypic multicellular spheroids (OMS) of glioma tissue were used as a culture model to study the

  12. Generation of multicellular tumor spheroids by the hanging-drop method.

    Science.gov (United States)

    Timmins, Nicholas E; Nielsen, Lars K

    2007-01-01

    Owing to their in vivo-like characteristics, three-dimensional (3D) multicellular tumor spheroid (MCTS) cultures are gaining increasing popularity as an in vitro model of tumors. A straightforward and simple approach to the cultivation of these MCTS is the hanging-drop method. Cells are suspended in droplets of medium, where they develop into coherent 3D aggregates and are readily accessed for analysis. In addition to being simple, the method eliminates surface interactions with an underlying substratum (e.g., polystyrene plastic or agarose), requires only a low number of starting cells, and is highly reproducible. This method has also been applied to the co-cultivation of mixed cell populations, including the co-cultivation of endothelial cells and tumor cells as a model of early tumor angiogenesis.

  13. Cyclooxygenase-2 inhibitor is a robust enhancer of anticancer agents against hepatocellular carcinoma multicellular spheroids

    Directory of Open Access Journals (Sweden)

    Cui J

    2014-02-01

    Full Text Available Jie Cui,1,2 Ya-Huan Guo,3 Hong-Yi Zhang,4 Li-Li Jiang,1 Jie-Qun Ma,1 Wen-Juan Wang,1 Min-Cong Wang,1 Cheng-Cheng Yang,1 Ke-Jun Nan,1 Li-Ping Song5 1Department of Oncology, First Affiliated Hospital, College of Medicine of Xi'an Jiaotong University, Xi'an, 2Department of Oncology, Yan'an University Affiliated Hospital, Yan'an, 3Department of Oncology, Shaanxi Province Cancer Hospital, Xi'an, 4Department of Urology, Yan'an University Affiliated Hospital, Yan'an, 5Department of Radiotherapy, First Affiliated Hospital, College of Medicine of Xi'an Jiaotong University, Xi'an, People's Republic of China Purpose: Celecoxib, an inhibitor of cyclooxygenase-2 (COX2, was investigated for enhancement of chemotherapeutic efficacy in cancer clinical trials. This study aimed to determine whether celecoxib combined with 5-fluorouracil or sorafenib or gefitinib is beneficial in HepG2 multicellular spheroids (MCSs, as well as elucidate the underlying mechanisms. Methods: The human hepatocellular carcinoma cell line HepG2 MCSs were used as in vitro models to investigate the effects of celecoxib combined with 5-fluorouracil or sorafenib or gefitinib treatment on cell growth, apoptosis, and signaling pathway. Results: MCSs showed resistance to drugs compared with monolayer cells. Celecoxib combined with 5-fluorouracil or sorafenib exhibited a synergistic action. Exposure to celecoxib (21.8 µmol/L plus 5-fluorouracil (8.1 × 10-3 g/L or sorafenib (4.4 µmol/L increased apoptosis but exerted no effect on COX2, phosphorylated epidermal growth-factor receptor (p-EGFR and phosphorylated (p-AKT expression. Gefitinib (5 µmol/L, which exhibits no growth-inhibition activity as a single agent, increased the inhibitory effect of celecoxib. Gefitinib (5 µmol/L plus celecoxib (21.8 µmol/L increased apoptosis. COX2, p-EGFR, and p-AKT were inhibited. Conclusion: Celecoxib combined with 5-fluorouracil or sorafenib or gefitinib may be superior to single-agent therapy in HepG2

  14. Low-temperature plasma-induced antiproliferative effects on multi-cellular tumor spheroids

    International Nuclear Information System (INIS)

    Plewa, Joseph-Marie; Yousfi, Mohammed; Eichwald, Olivier; Merbahi, Nofel; Frongia, Céline; Ducommun, Bernard; Lobjois, Valérie

    2014-01-01

    Biomedical applications of low-temperature plasmas are of growing interest, especially in the field of plasma-induced anti-tumor effects. The present work is aimed at investigating the regionalized antiproliferative effects of low-temperature plasmas on a multicellular tumor spheroid (MCTS), a model that mimics the 3D organization and regionalization of a microtumor region. We report that a low-temperature plasma jet, using helium flow in open air, inhibits HCT116 colon carcinoma MCTS growth in a dose-dependent manner. This growth inhibition is associated with the loss of Ki67, and the regionalized accumulation of DNA damage detected by histone H2AX phosphorylation. This regionalized genotoxic effect leads to massive cell death and loss of the MCTS proliferative region. The use of reactive oxygen species (ROS), scavenger N-acetyl cysteine (NAC) and plasma-conditioned media demonstrate that the ROS generated in the media after exposure to low-temperature plasma play a major role in these observed effects. These findings strengthen the interest in the use of MCTS for the evaluation of antiproliferative strategies, and open new perspectives for studies dedicated to demonstrate the potential of low-temperature plasma in cancer therapy

  15. Low-temperature plasma-induced antiproliferative effects on multi-cellular tumor spheroids

    Science.gov (United States)

    Plewa, Joseph-Marie; Yousfi, Mohammed; Frongia, Céline; Eichwald, Olivier; Ducommun, Bernard; Merbahi, Nofel; Lobjois, Valérie

    2014-04-01

    Biomedical applications of low-temperature plasmas are of growing interest, especially in the field of plasma-induced anti-tumor effects. The present work is aimed at investigating the regionalized antiproliferative effects of low-temperature plasmas on a multicellular tumor spheroid (MCTS), a model that mimics the 3D organization and regionalization of a microtumor region. We report that a low-temperature plasma jet, using helium flow in open air, inhibits HCT116 colon carcinoma MCTS growth in a dose-dependent manner. This growth inhibition is associated with the loss of Ki67, and the regionalized accumulation of DNA damage detected by histone H2AX phosphorylation. This regionalized genotoxic effect leads to massive cell death and loss of the MCTS proliferative region. The use of reactive oxygen species (ROS), scavenger N-acetyl cysteine (NAC) and plasma-conditioned media demonstrate that the ROS generated in the media after exposure to low-temperature plasma play a major role in these observed effects. These findings strengthen the interest in the use of MCTS for the evaluation of antiproliferative strategies, and open new perspectives for studies dedicated to demonstrate the potential of low-temperature plasma in cancer therapy.

  16. On the surviving fraction in irradiated multicellular tumour spheroids: calculation of overall radiosensitivity parameters, influence of hypoxia and volume effects

    International Nuclear Information System (INIS)

    Horas, Jorge A; Olguin, Osvaldo R; Rizzotto, Marcos G

    2005-01-01

    We model the heterogeneous response to radiation of multicellular tumour spheroids assuming position- and volume-dependent radiosensitivity. We propose a method to calculate the overall radiosensitivity parameters to obtain the surviving fraction of tumours. A mathematical model of a spherical tumour with a hypoxic core and a viable rim which is a caricature of a real tumour is constructed. The model is embedded in a two-compartment linear-quadratic (LQ) model, assuming a mixed bivariated Gaussian distribution to attain the radiosensitivity parameters. Ergodicity, i.e., the equivalence between ensemble and volumetric averages is used to obtain the overall radiosensitivities for the two compartments. We obtain expressions for the overall radiosensitivity parameters resulting from the use of both a linear and a nonlinear dependence of the local radiosensitivity with position. The model's results are compared with experimental data of surviving fraction (SF) for multicellular spheroids of different sizes. We make one fit using only the smallest spheroid data and we are able to predict the SF for the larger spheroids. These predictions are acceptable particularly using bounded sensitivities. We conclude with the importance of taking into account the contribution of clonogenic hypoxic cells to radiosensitivity and with the convenience of using bounded local sensitivities to predict overall radiosensitivity parameters

  17. Development of three-dimensional lung multicellular spheroids in air- and liquid-interface culture for the evaluation of anticancer therapeutics.

    Science.gov (United States)

    Meenach, Samantha A; Tsoras, Alexandra N; McGarry, Ronald C; Mansour, Heidi M; Hilt, J Zach; Anderson, Kimberly W

    2016-04-01

    Three-dimensional (3D) lung multicellular spheroids (MCS) in liquid-covered culture (LCC) and air-interface culture (AIC) conditions have both been developed for the evaluation of aerosol anticancer therapeutics in solution and aerosols, respectively. The MCS were formed by seeding lung cancer cells on top of collagen where they formed spheroids due to the prevalence of cell-to-cell interactions. LCC MCS were exposed to paclitaxel (PTX) in media whereas AIC MCS were exposed to dry powder PEGylated phospholipid aerosol microparticles containing paclitaxel. The difference in viability for 2D versus 3D culture for both LCC and AIC was evaluated along with the effects of the particles on lung epithelium via transepithelial electrical resistance (TEER) measurements. For LCC and AIC conditions, the 3D spheroids were more resistant to treatment with higher IC50 values for A549 and H358 cell lines. TEER results initially indicated a decrease in resistance upon drug or particle exposure, however, these values increased over the course of several days indicating the ability of the cells to recover. Overall, these studies offer a comprehensive in vitro evaluation of aerosol particles used in the treatment of lung cancer while introducing a new method for culturing lung cancer MCS in both LCC and AIC conditions.

  18. Chemosensitivity of irradiated resistant cells of multicellular spheroids in A549 lung adenocarcinoma

    International Nuclear Information System (INIS)

    Shi Degang; Shi Genming; Huang Gang

    2006-01-01

    Objective: To investigate the chemosensitivity of irradiated resistant cells of multicellular spheroids in A549 lung adenocarcinoma. Methods: The A549 irradiated resistant cells were the 10th regrowth generations after irradiated with 2.5 Gy of 6 MV X-ray, the control groups were A549 parent cells and MCFY/VCR resistant cells. The 6 kinds of chemotherapeutic drugs were DDP, VDS, 5-FU, HCP, MMC and ADM respectively, with verapamil (VPL) as reverse agent. The treatment effect was compared with MTT assay, and the multidrug resistant gene expressions of mdrl and MRP were measured with RT-PCR method. Results: A549 cells and irradiated resistant cells were resistant to DDP, but sensitivity to VDS,5-FU, HCP, MMC and ADM. The inhibitory rates of VPL to the above two cells were 98% and 25% respectively(P 2 -MG and MRP/β 2 -MG of all A549 cells were about 0 and 0.7 respectively, and those of MCFT/VCR cells were 35 and 4.36. Conclusion: The chemosensitivity of A549 irradiated resistant cells had not changed markedly, the decreased sensitivity to VPL could not be explained by the gene expression of mdrl and MRP. It is conferred that some kinds of changes in the cell membrane and decreased regrowth ability to result in resistance. Unlike multidrug resistance induced by chemotherapy, VPL may be not an ideal reverser to irradiated resistant cells. The new kinds of biological preparation should be sought to combine chemotherapy to treat recurring tumor with irradiated resistance. (authors)

  19. Superparamagnetic iron oxide nanoparticles exert different cytotoxic effects on cells grown in monolayer cell culture versus as multicellular spheroids

    Energy Technology Data Exchange (ETDEWEB)

    Theumer, Anja; Gräfe, Christine; Bähring, Franziska [Department of Hematology and Oncology, Jena University Hospital, Erlanger Allee 101, 07747 Jena (Germany); Bergemann, Christian [Chemicell GmbH, Eresburgstrasse 22–23, 12103 Berlin (Germany); Hochhaus, Andreas [Department of Hematology and Oncology, Jena University Hospital, Erlanger Allee 101, 07747 Jena (Germany); Clement, Joachim H., E-mail: joachim.clement@med.uni-jena.de [Department of Hematology and Oncology, Jena University Hospital, Erlanger Allee 101, 07747 Jena (Germany)

    2015-04-15

    The aim of this study was to investigate the interaction of superparamagnetic iron oxide nanoparticles (SPION) with human blood–brain barrier-forming endothelial cells (HBMEC) in two-dimensional cell monolayers as well as in three-dimensional multicellular spheroids. The precise nanoparticle localisation and the influence of the NP on the cellular viability and the intracellular Akt signalling were studied in detail. Long-term effects of different polymer-coated nanoparticles (neutral fluidMAG-D, anionic fluidMAG-CMX and cationic fluidMAG-PEI) and the corresponding free polymers on cellular viability of HBMEC were investigated by real time cell analysis studies. Nanoparticles exert distinct effects on HBMEC depending on the nanoparticles' surface charge and concentration, duration of incubation and cellular context. The most severe effects were caused by PEI-coated nanoparticles. Concentrations above 25 µg/ml led to increased amounts of dead cells in monolayer culture as well as in multicellular spheroids. On the level of intracellular signalling, context-dependent differences were observed. Monolayer cultures responded on nanoparticle incubation with an increase in Akt phosphorylation whereas spheroids on the whole show a decreased Akt activity. This might be due to the differential penetration and distribution of PEI-coated nanoparticles.

  20. Cell-like pressure sensors reveal increase of mechanical stress towards the core of multicellular spheroids under compression.

    Science.gov (United States)

    Dolega, M E; Delarue, M; Ingremeau, F; Prost, J; Delon, A; Cappello, G

    2017-01-27

    The surrounding microenvironment limits tumour expansion, imposing a compressive stress on the tumour, but little is known how pressure propagates inside the tumour. Here we present non-destructive cell-like microsensors to locally quantify mechanical stress distribution in three-dimensional tissue. Our sensors are polyacrylamide microbeads of well-defined elasticity, size and surface coating to enable internalization within the cellular environment. By isotropically compressing multicellular spheroids (MCS), which are spherical aggregates of cells mimicking a tumour, we show that the pressure is transmitted in a non-trivial manner inside the MCS, with a pressure rise towards the core. This observed pressure profile is explained by the anisotropic arrangement of cells and our results suggest that such anisotropy alone is sufficient to explain the pressure rise inside MCS composed of a single cell type. Furthermore, such pressure distribution suggests a direct link between increased mechanical stress and previously observed lack of proliferation within the spheroids core.

  1. Solid freeform-fabricated scaffolds designed to carry multicellular mesenchymal stem cell spheroids for cartilage regeneration

    Directory of Open Access Journals (Sweden)

    G-S Huang

    2013-10-01

    Full Text Available Three-dimensional (3D cellular spheroids have recently emerged as a new trend to replace suspended single cells in modern cell-based therapies because of their greater regeneration capacities in vitro. They may lose the 3D structure during a change of microenvironment, which poses challenges to their translation in vivo. Besides, the conventional microporous scaffolds may have difficulty in accommodating these relatively large spheroids. Here we revealed a novel design of microenvironment for delivering and sustaining the 3D spheroids. Biodegradable scaffolds with macroporosity to accommodate mesenchymal stem cell (MSC spheroids were made by solid freeform fabrication (SFF from the solution of poly(D,L-lactide-co-glycolide. Their internal surface was modified with chitosan following air plasma treatment in order to preserve the morphology of the spheroids. It was demonstrated that human MSC spheroids loaded in SFF scaffolds produced a significantly larger amount of cartilage-associated extracellular matrix in vitro and in NOD/SCID mice compared to single cells in the same scaffolds. Implantation of MSC spheroid-loaded scaffolds into the chondral defects of rabbit knees showed superior cartilage regeneration. This study establishes new perspectives in designing the spheroid-sustaining microenvironment within a tissue engineering scaffold for in vivo applications.

  2. Treatment Efficiency of Free and Nanoparticle-Loaded Mitoxantrone for Magnetic Drug Targeting in Multicellular Tumor Spheroids

    Directory of Open Access Journals (Sweden)

    Annkathrin Hornung

    2015-09-01

    Full Text Available Major problems of cancer treatment using systemic chemotherapy are severe side effects. Magnetic drug targeting (MDT employing superparamagnetic iron oxide nanoparticles (SPION loaded with chemotherapeutic agents may overcome this dilemma by increasing drug accumulation in the tumor and reducing toxic side effects in the healthy tissue. For translation of nanomedicine from bench to bedside, nanoparticle-mediated effects have to be studied carefully. In this study, we compare the effect of SPION, unloaded or loaded with the cytotoxic drug mitoxantrone (MTO with the effect of free MTO, on the viability and proliferation of HT-29 cells within three-dimensional multicellular tumor spheroids. Fluorescence microscopy and flow cytometry showed that both free MTO, as well as SPION-loaded MTO (SPIONMTO are able to penetrate into tumor spheroids and thereby kill tumor cells, whereas unloaded SPION did not affect cellular viability. Since SPIONMTO has herewith proven its effectivity also in complex multicellular tumor structures with its surrounding microenvironment, we conclude that it is a promising candidate for further use in magnetic drug targeting in vivo.

  3. Molecular and functional assessment of multicellular cancer spheroids produced in double emulsions enabled by efficient airway resistance based selective surface treatment

    Science.gov (United States)

    Ma, Xiao; Leth Jepsen, Morten; Ivarsen, Anne Kathrine R.; Knudsen, Birgitta R.; Ho, Yi-Ping

    2017-09-01

    Multicellular spheroids have garnered significant attention as an in vitro three-dimensional cancer model which can mimick the in vivo microenvironmental features. While microfluidics generated double emulsions have become a potential method to generate spheroids, challenges remain on the tedious procedures. Enabled by a novel ‘airway resistance’ based selective surface treatment, this study presents an easy and facile generation of double emulsions for the initiation and cultivation of multicellular spheroids in a scaffold-free format. Combining with our previously developed DNA nanosensors, intestinal spheroids produced in the double emulsions have shown an elevated activities of an essential DNA modifying enzyme, the topoisomerase I. The observed molecular and functional characteristics of spheroids produced in double emulsions are similar to the counterparts produced by the commercially available ultra-low attachment plates. However, the double emulsions excel for their improved uniformity, and the consistency of the results obtained by subsequent analysis of the spheroids. The presented technique is expected to ease the burden of producing spheroids and to promote the spheroids model for cancer or stem cell study.

  4. Optimization of the formation of embedded multicellular spheroids of MCF-7 cells: How to reliably produce a biomimetic 3D model.

    Science.gov (United States)

    Zhang, Wenli; Li, Caibin; Baguley, Bruce C; Zhou, Fang; Zhou, Weisai; Shaw, John P; Wang, Zhen; Wu, Zimei; Liu, Jianping

    2016-12-15

    To obtain a multicellular MCF-7 spheroid model to mimic the three-dimensional (3D) of tumors, the microwell liquid overlay (A) and hanging-drop/agar (B) methods were first compared for their technical parameters. Then a method for embedding spheroids within collagen was optimized. For method A, centrifugation assisted cells form irregular aggregates but not spheroids. For method B, an extended sedimentation period of over 24 h for cell suspensions and increased viscosity of the culture medium using methylcellulose were necessary to harvest a dense and regular cell spheroid. When the number was less than 5000 cells/drop, embedded spheroids showed no tight cores and higher viability than the unembedded. However, above 5000 cells/drop, cellular viability of embedded spheroids was not significantly different from unembedded spheroids and cells invading through the collagen were in a sun-burst pattern with tight cores. Propidium Iodide staining indicated that spheroids had necrotic cores. The doxorubicin cytotoxicity demonstrated that spheroids were less susceptible to DOX than their monolayer cells. A reliable and reproducible method for embedding spheroids using the hanging-drop/agarose method within collagen is described herein. The cell culture model can be used to guide experimental manipulation of 3D cell cultures and to evaluate anticancer drug efficacy. Copyright © 2016 Elsevier Inc. All rights reserved.

  5. Hypoxic fraction and binding of misonidazole in EMT6/Ed multicellular tumor spheroids

    International Nuclear Information System (INIS)

    Franko, A.J.

    1985-01-01

    Misonidazole has been shown to bind selectively to hypoxic cells in tissue culture and to cells which are presumed to be chronically hypoxic in EMT6 spheroids and tumors. Thus it has considerable potential as a marker of hypoxic cells in vivo. To further evaluate this potential EMT6/Ed spheroids were used to quantitate misonidazole binding under conditions which resulted in hypoxic fractions between 0 and 1. The patterns of binding of 14 C-labeled misonidazole determined by autoradiography were consistent with the regions of radiobiological hypoxia as predicted by oxygen diffusion theory. The overall uptake of 3 H-labeled misonidazole by spheroids correlated well with the hypoxic fraction, although binding to aerobic cells and necrotic tissue contributed appreciably to the total label in the spheroids. It is concluded that misonidazole is an excellent marker of hypoxia in EMT6/Ed spheroids at the microscopic level, and the total amount bound per spheroid provides a potentially useful measure of the hypoxic fraction

  6. Development of complex-shaped liver multicellular spheroids as a human-based model for nanoparticle toxicity assessment in vitro

    International Nuclear Information System (INIS)

    Dubiak-Szepietowska, Monika; Karczmarczyk, Aleksandra; Jönsson-Niedziółka, Martin; Winckler, Thomas; Feller, Karl-Heinz

    2016-01-01

    The emergence of human-based models is incontestably required for the study of complex physiological pathways and validation of reliable in vitro methods as alternative for in vivo studies in experimental animals for toxicity assessment. With this objective, we have developed and tested three dimensional environments for cells using different types of hydrogels including transglutaminase-cross-linked gelatin, collagen type I, and growth-factor depleted Matrigel. Cells grown in Matrigel exhibited the greatest cell proliferation and spheroid diameter. Moreover, analysis of urea and albumin biosynthesis revealed that the created system allowed the immortalized liver cell line HepG2 to re-establish normal hepatocyte-like properties which were not observed under the conditions of conventional cell cultures. This study presents a scalable technology for production of complex-shaped liver multicellular spheroids as a system which improves the predictive value of cell-based assays for safety and risk assessment. The time- and dose-dependent toxicity of nanoparticles demonstrates a higher cytotoxic effect when HepG2 cells grown as monolayer than embedded in hydrogels. The experimental setup provided evidence that the cell environment has significant influence on cell sensitivity and that liver spheroid is a useful and novel tool to examine nanoparticle dosing effect even at the level of in vitro studies. Therefore, this system can be applied to a wide variety of potentially hostile compounds in basic screening to provide initial warning of adverse effects and trigger subsequent analysis and remedial actions. - Highlights: • Comparison of HepG2 cells growth in Matrigel, Collagen I gel and gelatin gel. • Examination of nanoparticles (NP) dosing effect at the level of in vitro studies. • Influence of the cell culture media composition on the cytotoxic effect of NP.

  7. Down-regulation of DNA mismatch repair enhances initiation and growth of neuroblastoma and brain tumour multicellular spheroids.

    Directory of Open Access Journals (Sweden)

    Samuel L Collins

    Full Text Available Multicellular tumour spheroid (MCTS cultures are excellent model systems for simulating the development and microenvironmental conditions of in vivo tumour growth. Many documented cell lines can generate differentiated MCTS when cultured in suspension or in a non-adhesive environment. While physiological and biochemical properties of MCTS have been extensively characterized, insight into the events and conditions responsible for initiation of these structures is lacking. MCTS are formed by only a small subpopulation of cells during surface-associated growth but the processes responsible for this differentiation are poorly understood and have not been previously studied experimentally. Analysis of gene expression within spheroids has provided clues but to date it is not known if the observed differences are a cause or consequence of MCTS growth. One mechanism linked to tumourigenesis in a number of cancers is genetic instability arising from impaired DNA mismatch repair (MMR. This study aimed to determine the role of MMR in MCTS initiation and development. Using surface-associated N2a and CHLA-02-ATRT culture systems we have investigated the impact of impaired MMR on MCTS growth. Analysis of the DNA MMR genes MLH1 and PMS2 revealed both to be significantly down-regulated at the mRNA level compared with non-spheroid-forming cells. By using small interfering RNA (siRNA against these genes we show that silencing of MLH1 and PMS2 enhances both MCTS initiation and subsequent expansion. This effect was prolonged over several passages following siRNA transfection. Down-regulation of DNA MMR can contribute to tumour initiation and progression in N2a and CHLA-02-ATRT MCTS models. Studies of surface-associated MCTS differentiation may have broader applications in studying events in the initiation of cancer foci.

  8. Development of complex-shaped liver multicellular spheroids as a human-based model for nanoparticle toxicity assessment in vitro

    Energy Technology Data Exchange (ETDEWEB)

    Dubiak-Szepietowska, Monika, E-mail: Monika.Dubiak-Szepietowska@fh-jena.de [Department of Medical Engineering and Biotechnology, Ernst-Abbe-University of Applied Sciences Jena, Carl-Zeiss Promenade 2, 07745 Jena (Germany); Karczmarczyk, Aleksandra [Department of Medical Engineering and Biotechnology, Ernst-Abbe-University of Applied Sciences Jena, Carl-Zeiss Promenade 2, 07745 Jena (Germany); Jönsson-Niedziółka, Martin [Institute of Physical Chemistry, Polish Academy of Sciences, ul. Kasprzaka 44/52, 01-224 Warszawa (Poland); Winckler, Thomas [Institute of Pharmacy, Friedrich Schiller University Jena, Semmelweissstraße 10, 07743 Jena (Germany); Feller, Karl-Heinz [Department of Medical Engineering and Biotechnology, Ernst-Abbe-University of Applied Sciences Jena, Carl-Zeiss Promenade 2, 07745 Jena (Germany)

    2016-03-01

    The emergence of human-based models is incontestably required for the study of complex physiological pathways and validation of reliable in vitro methods as alternative for in vivo studies in experimental animals for toxicity assessment. With this objective, we have developed and tested three dimensional environments for cells using different types of hydrogels including transglutaminase-cross-linked gelatin, collagen type I, and growth-factor depleted Matrigel. Cells grown in Matrigel exhibited the greatest cell proliferation and spheroid diameter. Moreover, analysis of urea and albumin biosynthesis revealed that the created system allowed the immortalized liver cell line HepG2 to re-establish normal hepatocyte-like properties which were not observed under the conditions of conventional cell cultures. This study presents a scalable technology for production of complex-shaped liver multicellular spheroids as a system which improves the predictive value of cell-based assays for safety and risk assessment. The time- and dose-dependent toxicity of nanoparticles demonstrates a higher cytotoxic effect when HepG2 cells grown as monolayer than embedded in hydrogels. The experimental setup provided evidence that the cell environment has significant influence on cell sensitivity and that liver spheroid is a useful and novel tool to examine nanoparticle dosing effect even at the level of in vitro studies. Therefore, this system can be applied to a wide variety of potentially hostile compounds in basic screening to provide initial warning of adverse effects and trigger subsequent analysis and remedial actions. - Highlights: • Comparison of HepG2 cells growth in Matrigel, Collagen I gel and gelatin gel. • Examination of nanoparticles (NP) dosing effect at the level of in vitro studies. • Influence of the cell culture media composition on the cytotoxic effect of NP.

  9. Radiation-biological investigations with multicellular spheroids as an in vitro tumour model. 2

    International Nuclear Information System (INIS)

    Koerner, I.J.; Kopp, J.; Malz, W.

    1978-01-01

    Using the alkaline sucrose gradient sedimentation technique, the single-strand break rejoining in single cells and cells of small sheroids was studied after X-irradiation with 10 krads. The half life time for the fast rejoining process amounts to about 4 minutes for single cells and to about 1.5 minutes for small spheroids. While in single cells 7 per cent of single-strand breaks remain unrepaired after 80 minutes incubation at 37 0 C, in small spheroids practically no residual unrepaired single-strand breaks can be detected after more than 20 minutes. The corresponding survival of cells of small sheroids is increased in comparison with single cells. Possible reasons are discussed for the different repair capacities found. (author)

  10. Visualizing the effect of tumor microenvironments on radiation-induced cell kinetics in multicellular spheroids consisting of HeLa cells

    International Nuclear Information System (INIS)

    Kaida, Atsushi; Miura, Masahiko

    2013-01-01

    Highlights: •We visualized radiation-induced cell kinetics in spheroids. •HeLa-Fucci cells were used for detection of cell-cycle changes. •Radiation-induced G2 arrest was prolonged in the spheroid. •The inner and outer cell fractions behaved differently. -- Abstract: In this study, we visualized the effect of tumor microenvironments on radiation-induced tumor cell kinetics. For this purpose, we utilized a multicellular spheroid model, with a diameter of ∼500 μm, consisting of HeLa cells expressing the fluorescent ubiquitination-based cell-cycle indicator (Fucci). In live spheroids, a confocal laser scanning microscope allowed us to clearly monitor cell kinetics at depths of up to 60 μm. Surprisingly, a remarkable prolongation of G2 arrest was observed in the outer region of the spheroid relative to monolayer-cultured cells. Scale, an aqueous reagent that renders tissues optically transparent, allowed visualization deeper inside spheroids. About 16 h after irradiation, a red fluorescent cell fraction, presumably a quiescent G0 cell fraction, became distinct from the outer fraction consisting of proliferating cells, most of which exhibited green fluorescence indicative of G2 arrest. Thereafter, the red cell fraction began to emit green fluorescence and remained in prolonged G2 arrest. Thus, for the first time, we visualized the prolongation of radiation-induced G2 arrest in spheroids and the differences in cell kinetics between the outer and inner fractions

  11. A microfluidic chip with a U-shaped microstructure array for multicellular spheroid formation, culturing and analysis

    International Nuclear Information System (INIS)

    Fu, Chien-Yu; Chang, Hwan-You; Tseng, Sheng-Yang; Yang, Shih-Mo; Hsu, Long; Liu, Cheng-Hsien

    2014-01-01

    Multicellular spheroids (MCS), formed by self-assembly of single cells, are commonly used as a three-dimensional cell culture model to bridge the gap between in vitro monolayer culture and in vivo tissues. However, current methods for MCS generation and analysis still suffer drawbacks such as being labor-intensive and of poor controllability, and are not suitable for high-throughput applications. This study demonstrates a novel microfluidic chip to facilitate MCS formation, culturing and analysis. The chip contains an array of U-shaped microstructures fabricated by photopolymerizing the poly(ethylene glycol) diacrylate hydrogel through defining the ultraviolet light exposure pattern with a photomask. The geometry of the U-shaped microstructures allowed trapping cells into the pocket through the actions of fluid flow and the force of gravity. The hydrogel is non-adherent for cells, promoting the formation of MCS. Its permselective property also facilitates exchange of nutrients and waste for MCS, while providing protection of MCS from shearing stress during the medium perfusion. Heterotypic MCS can be formed easily by manipulating the cell trapping steps. Subsequent drug susceptibility analysis and long-term culture could also be achieved within the same chip. This MCS formation and culture platform can be used as a micro-scale bioreactor and applied in many cell biology and drug testing studies. (paper)

  12. Hypofractionated radiation induces a decrease in cell proliferation but no histological damage to organotypic multicellular spheroids of human glioblastomas

    International Nuclear Information System (INIS)

    Kaaijk, P.; Academic Medical Center, Amsterdam; Troost, D.; Leenstra, S.; Bosch, D.A.; Sminia, P.; Hulshof, M.C.C.M..; Kracht, A.H.W. van der

    1997-01-01

    The aim of this study was to examine the effect of radiation on glioblastoma, using an organotypic multicellular spheroid (OMS) model. Most glioblastoma cell lines are, in contrast to glioblastomas in vivo, relatively radiosensitive. This limits the value of using cell lines for studying the radiation effect of glioblastomas. The advantage of OMS is maintenance of the characteristics of the original tumour, which is lost in conventional cell cultures. OMS prepared from four glioblastomas were treated with hypofractionated radiation with a radiobiologically equivalent dose to standard radiation treatment for glioblastomas patients. After treatment, the histology as well as the cell proliferation of the OMS was examined. After radiation, a significant decrease in cell proliferation was found, although no histological damage to the OMS was observed. The modest effects of radiation on the OMS are in agreement with the limited therapeutic value of radiotherapy for glioblastoma patients. Therefore, OMS seems to be a good alternative for cell lines to study the radiobiological effect on glioblastomas. (author)

  13. Hypofractionated radiation induces a decrease in cell proliferation but no histological damage to organotypic multicellular spheroids of human glioblastomas

    Energy Technology Data Exchange (ETDEWEB)

    Kaaijk, P [Academic Medical Center, Amsterdam (Netherlands). Dept. of (Neuro) Pathology; [Academic Medical Center, Amsterdam (Netherlands). Dept. of Neurosurgery; Troost, D [Academic Medical Center, Amsterdam (Netherlands). Dept. of (Neuro) Pathology; Leenstra, S; Bosch, D A [Academic Medical Center, Amsterdam (Netherlands). Dept. of Neurosurgery; Sminia, P; Hulshof, M C.C.M.; Kracht, A.H.W. van der [Academic Medical Center, Amsterdam (Netherlands). Dept. of (Experimental) Radiotherapy

    1997-04-01

    The aim of this study was to examine the effect of radiation on glioblastoma, using an organotypic multicellular spheroid (OMS) model. Most glioblastoma cell lines are, in contrast to glioblastomas in vivo, relatively radiosensitive. This limits the value of using cell lines for studying the radiation effect of glioblastomas. The advantage of OMS is maintenance of the characteristics of the original tumour, which is lost in conventional cell cultures. OMS prepared from four glioblastomas were treated with hypofractionated radiation with a radiobiologically equivalent dose to standard radiation treatment for glioblastomas patients. After treatment, the histology as well as the cell proliferation of the OMS was examined. After radiation, a significant decrease in cell proliferation was found, although no histological damage to the OMS was observed. The modest effects of radiation on the OMS are in agreement with the limited therapeutic value of radiotherapy for glioblastoma patients. Therefore, OMS seems to be a good alternative for cell lines to study the radiobiological effect on glioblastomas. (author).

  14. Short and long time effects of low temperature Plasma Activated Media on 3D multicellular tumor spheroids

    Science.gov (United States)

    Judée, Florian; Fongia, Céline; Ducommun, Bernard; Yousfi, Mohammed; Lobjois, Valérie; Merbahi, Nofel

    2016-02-01

    This work investigates the regionalized antiproliferative effects of plasma-activated medium (PAM) on colon adenocarcinoma multicellular tumor spheroid (MCTS), a model that mimics 3D organization and regionalization of a microtumor region. PAM was generated by dielectric barrier plasma jet setup crossed by helium carrier gas. MCTS were transferred in PAM at various times after plasma exposure up to 48 hours and effect on MCTS growth and DNA damage were evaluated. We report the impact of plasma exposure duration and delay before transfer on MCTS growth and DNA damage. Local accumulation of DNA damage revealed by histone H2AX phosphorylation is observed on outermost layers and is dependent on plasma exposure. DNA damage is completely reverted by catalase addition indicating that H2O2 plays major role in observed genotoxic effect while growth inhibitory effect is maintained suggesting that it is due to others reactive species. SOD and D-mannitol scavengers also reduced DNA damage by 30% indicating that and OH* are involved in H2O2 formation. Finally, PAM is able to retain its cytotoxic and genotoxic activity upon storage at +4 °C or -80 °C. These results suggest that plasma activated media may be a promising new antitumor strategy for colorectal cancer tumors.

  15. Doxorubicin delivery to 3D multicellular spheroids and tumors based on boronic acid-rich chitosan nanoparticles.

    Science.gov (United States)

    Wang, Xin; Zhen, Xu; Wang, Jing; Zhang, Jialiang; Wu, Wei; Jiang, Xiqun

    2013-06-01

    Boronic acid-rich chitosan-poly(N-3-acrylamidophenylboronic acid) nanoparticles (CS-PAPBA NPs) with the tunable size were successfully prepared by polymerizing N-3-acrylamidophenylboronic acid in the presence of chitosan in an aqueous solution. The CS-PAPBA NPs were then functionalized by a tumor-penetrating peptide iRGD and loading doxorubicin (DOX). The interaction between boronic acid groups of hydrophobic PAPBA and the amino groups of hydrophilic chitosan inside the nanoparticles was examined by solid-state NMR measurement. The size and morphology of nanoparticles were characterized by dynamic light scattering and electron microscopy. The cellular uptake, tumor penetration, biodistribution and antitumor activity of the nanoparticles were evaluated by using three-dimensional (3-D) multicellular spheroids (MCs) as the in vitro model and H22 tumor-bearing mice as the in vivo model. It was found that the iRGD-conjugated nanoparticles significantly improved the efficiency of DOX penetration in MCs, compared with free DOX and non-conjugated nanoparticles, resulting in the efficient cell killing in the MCs. In vivo antitumor activity examination indicated that iRGD-conjugated CS-PAPBA nanoparticles promoted the accumulation of nanoparticles in tumor tissue and enhanced their penetration in tumor areas, both of which improved the efficiency of DOX-loaded nanoparticles in restraining tumor growth and prolonging the life time of H22 tumor-bearing mice. Copyright © 2013 Elsevier Ltd. All rights reserved.

  16. Drug delivery to solid tumors: the predictive value of the multicellular tumor spheroid model for nanomedicine screening

    Directory of Open Access Journals (Sweden)

    Millard M

    2017-10-01

    Full Text Available Marie Millard,1,2 Ilya Yakavets,1–3 Vladimir Zorin,3,4 Aigul Kulmukhamedova,1,2,5 Sophie Marchal,1,2 Lina Bezdetnaya1,2 1Centre de Recherche en Automatique de Nancy, Centre National de la Recherche Scientifique UMR 7039, Université de Lorraine, 2Research Department, Institut de Cancérologie de Lorraine, Vandœuvre-lès-Nancy, France; 3Laboratory of Biophysics and Biotechnology, 4International Sakharov Environmental Institute, Belarusian State University, Minsk, Belarus; 5Department of Radiology, Medical Company Sunkar, Almaty, Kazakhstan Abstract: The increasing number of publications on the subject shows that nanomedicine is an attractive field for investigations aiming to considerably improve anticancer chemotherapy. Based on selective tumor targeting while sparing healthy tissue, carrier-mediated drug delivery has been expected to provide significant benefits to patients. However, despite reduced systemic toxicity, most nanodrugs approved for clinical use have been less effective than previously anticipated. The gap between experimental results and clinical outcomes demonstrates the necessity to perform comprehensive drug screening by using powerful preclinical models. In this context, in vitro three-dimensional models can provide key information on drug behavior inside the tumor tissue. The multicellular tumor spheroid (MCTS model closely mimics a small avascular tumor with the presence of proliferative cells surrounding quiescent cells and a necrotic core. Oxygen, pH and nutrient gradients are similar to those of solid tumor. Furthermore, extracellular matrix (ECM components and stromal cells can be embedded in the most sophisticated spheroid design. All these elements together with the physicochemical properties of nanoparticles (NPs play a key role in drug transport, and therefore, the MCTS model is appropriate to assess the ability of NP to penetrate the tumor tissue. This review presents recent developments in MCTS models for a

  17. Time-lapse 3-D measurements of a glucose biosensor in multicellular spheroids by light sheet fluorescence microscopy in commercial 96-well plates.

    Science.gov (United States)

    Maioli, Vincent; Chennell, George; Sparks, Hugh; Lana, Tobia; Kumar, Sunil; Carling, David; Sardini, Alessandro; Dunsby, Chris

    2016-11-25

    Light sheet fluorescence microscopy has previously been demonstrated on a commercially available inverted fluorescence microscope frame using the method of oblique plane microscopy (OPM). In this paper, OPM is adapted to allow time-lapse 3-D imaging of 3-D biological cultures in commercially available glass-bottomed 96-well plates using a stage-scanning OPM approach (ssOPM). Time-lapse 3-D imaging of multicellular spheroids expressing a glucose Förster resonance energy transfer (FRET) biosensor is demonstrated in 16 fields of view with image acquisition at 10 minute intervals. As a proof-of-principle, the ssOPM system is also used to acquire a dose response curve with the concentration of glucose in the culture medium being varied across 42 wells of a 96-well plate with the whole acquisition taking 9 min. The 3-D image data enable the FRET ratio to be measured as a function of distance from the surface of the spheroid. Overall, the results demonstrate the capability of the OPM system to measure spatio-temporal changes in FRET ratio in 3-D in multicellular spheroids over time in a multi-well plate format.

  18. Developing multi-cellular tumor spheroid model (MCTS) in the chitosan/collagen/alginate (CCA) fibrous scaffold for anticancer drug screening

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Jian-Zheng, E-mail: wppzheng@126.com [Laboratory of Biomedical Material Engineering, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, 457 Zhongshan Road, Dalian 116023 (China); Affiliated General Hospital, Tianguan Group Co., Ltd, Nanyang 473000 (China); Testing Center of Henan Tianguan Group Co., Ltd, Nanyang 473000 (China); Zhu, Yu-Xia [Laboratory of Biomedical Material Engineering, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, 457 Zhongshan Road, Dalian 116023 (China); Affiliated General Hospital, Tianguan Group Co., Ltd, Nanyang 473000 (China); Testing Center of Henan Tianguan Group Co., Ltd, Nanyang 473000 (China); Ma, Hui-Chao; Chen, Si-Nan; Chao, Ji-Ye; Ruan, Wen-Ding; Wang, Duo; Du, Feng-guang [Affiliated General Hospital, Tianguan Group Co., Ltd, Nanyang 473000 (China); Testing Center of Henan Tianguan Group Co., Ltd, Nanyang 473000 (China); Meng, Yue-Zhong [State Key Laboratory of Optoelectronic Materials and Technologies, Sun Yat-sen University, Guangzhou 510275 (China)

    2016-05-01

    In this work, a 3D MCTS-CCA system was constructed by culturing multi-cellular tumor spheroid (MCTS) in the chitosan/collagen/alginate (CCA) fibrous scaffold for anticancer drug screening. The CCA scaffolds were fabricated by spray-spinning. The interactions between the components of the spray-spun fibers were evidenced by methods of Coomassie Blue stain, X-ray diffraction (XRD) and Fourier transform-infrared spectroscopy (FTIR). Co-culture indicated that MCF-7 cells showed a spatial growth pattern of multi-cellular tumor spheroid (MCTS) in the CCA fibrous scaffold with increased proliferation rate and drug-resistance to MMC, ADM and 5-Aza comparing with the 2D culture cells. Significant increases of total viable cells were found in 3D MCTS groups after drug administration by method of apoptotic analysis. Glucose–lactate analysis indicated that the metabolism of MCTS in CCA scaffold was closer to the tumor issue in vivo than the monolayer cells. In addition, MCTS showed the characteristic of epithelial mesenchymal transition (EMT) which is subverted by carcinoma cells to facilitate metastatic spread. These results demonstrated that MCTS in CCA scaffold possessed a more conservative phenotype of tumor than monolayer cells, and anticancer drug screening in 3D MCTS-CCA system might be superior to the 2D culture system. - Highlights: • Chitosan/collagen/alginate (CCA) scaffolds were fabricated by spray-spinning. • MCF-7 cells presented a multi-cellular tumor spheroid model (MCTS) in CCA scaffold. • MCTS in CCA possessed a more conservative phenotype of tumor than monolayer cells. • Anticancer drug screening in MCTS-CCA system is superior to 2D culture system.

  19. Developing multi-cellular tumor spheroid model (MCTS) in the chitosan/collagen/alginate (CCA) fibrous scaffold for anticancer drug screening

    International Nuclear Information System (INIS)

    Wang, Jian-Zheng; Zhu, Yu-Xia; Ma, Hui-Chao; Chen, Si-Nan; Chao, Ji-Ye; Ruan, Wen-Ding; Wang, Duo; Du, Feng-guang; Meng, Yue-Zhong

    2016-01-01

    In this work, a 3D MCTS-CCA system was constructed by culturing multi-cellular tumor spheroid (MCTS) in the chitosan/collagen/alginate (CCA) fibrous scaffold for anticancer drug screening. The CCA scaffolds were fabricated by spray-spinning. The interactions between the components of the spray-spun fibers were evidenced by methods of Coomassie Blue stain, X-ray diffraction (XRD) and Fourier transform-infrared spectroscopy (FTIR). Co-culture indicated that MCF-7 cells showed a spatial growth pattern of multi-cellular tumor spheroid (MCTS) in the CCA fibrous scaffold with increased proliferation rate and drug-resistance to MMC, ADM and 5-Aza comparing with the 2D culture cells. Significant increases of total viable cells were found in 3D MCTS groups after drug administration by method of apoptotic analysis. Glucose–lactate analysis indicated that the metabolism of MCTS in CCA scaffold was closer to the tumor issue in vivo than the monolayer cells. In addition, MCTS showed the characteristic of epithelial mesenchymal transition (EMT) which is subverted by carcinoma cells to facilitate metastatic spread. These results demonstrated that MCTS in CCA scaffold possessed a more conservative phenotype of tumor than monolayer cells, and anticancer drug screening in 3D MCTS-CCA system might be superior to the 2D culture system. - Highlights: • Chitosan/collagen/alginate (CCA) scaffolds were fabricated by spray-spinning. • MCF-7 cells presented a multi-cellular tumor spheroid model (MCTS) in CCA scaffold. • MCTS in CCA possessed a more conservative phenotype of tumor than monolayer cells. • Anticancer drug screening in MCTS-CCA system is superior to 2D culture system.

  20. Three-dimensional growth as multicellular spheroid activates the proangiogenic phenotype of colorectal carcinoma cells via LFA-1-dependent VEGF: implications on hepatic micrometastasis

    Directory of Open Access Journals (Sweden)

    Muruzabal Francisco J

    2008-10-01

    Full Text Available Abstract Background The recruitment of vascular stromal and endothelial cells is an early event occurring during cancer cell growth at premetastatic niches, but how the microenvironment created by the initial three-dimensional (3D growth of cancer cells affects their angiogenesis-stimulating potential is unclear. Methods The proangiogenic profile of CT26 murine colorectal carcinoma cells was studied in seven-day cultured 3D-spheroids of Results Spheroid-derived CT26 cells increased vascular endothelial growth factor (VEGF secretion by 70%, which in turn increased the in vitro migration of primary cultured hepatic sinusoidal endothelium (HSE cells by 2-fold. More importantly, spheroid-derived CT26 cells increased lymphocyte function associated antigen (LFA-1-expressing cell fraction by 3-fold; and soluble intercellular adhesion molecule (ICAM-1, given to spheroid-cultured CT26 cells, further increased VEGF secretion by 90%, via cyclooxygenase (COX-2-dependent mechanism. Consistent with these findings, CT26 cancer cells significantly increased LFA-1 expression in non-hypoxic avascular micrometastases at their earliest inception within hepatic lobules in vivo; and angiogenesis also markedly increased in both subcutaneous tumors and hepatic metastases produced by spheroid-derived CT26 cells. Conclusion 3D-growth per se enriched the proangiogenic phenotype of cancer cells growing as multicellular spheroids or as subclinical hepatic micrometastases. The contribution of integrin LFA-1 to VEGF secretion via COX-2 was a micro environmental-related mechanism leading to the pro-angiogenic activation of soluble ICAM-1-activated colorectal carcinoma cells. This mechanism may represent a new target for specific therapeutic strategies designed to block colorectal cancer cell growth at a subclinical micrometastatic stage within the liver.

  1. Inferring Growth Control Mechanisms in Growing Multi-cellular Spheroids of NSCLC Cells from Spatial-Temporal Image Data.

    Science.gov (United States)

    Jagiella, Nick; Müller, Benedikt; Müller, Margareta; Vignon-Clementel, Irene E; Drasdo, Dirk

    2016-02-01

    We develop a quantitative single cell-based mathematical model for multi-cellular tumor spheroids (MCTS) of SK-MES-1 cells, a non-small cell lung cancer (NSCLC) cell line, growing under various nutrient conditions: we confront the simulations performed with this model with data on the growth kinetics and spatial labeling patterns for cell proliferation, extracellular matrix (ECM), cell distribution and cell death. We start with a simple model capturing part of the experimental observations. We then show, by performing a sensitivity analysis at each development stage of the model that its complexity needs to be stepwise increased to account for further experimental growth conditions. We thus ultimately arrive at a model that mimics the MCTS growth under multiple conditions to a great extent. Interestingly, the final model, is a minimal model capable of explaining all data simultaneously in the sense, that the number of mechanisms it contains is sufficient to explain the data and missing out any of its mechanisms did not permit fit between all data and the model within physiological parameter ranges. Nevertheless, compared to earlier models it is quite complex i.e., it includes a wide range of mechanisms discussed in biological literature. In this model, the cells lacking oxygen switch from aerobe to anaerobe glycolysis and produce lactate. Too high concentrations of lactate or too low concentrations of ATP promote cell death. Only if the extracellular matrix density overcomes a certain threshold, cells are able to enter the cell cycle. Dying cells produce a diffusive growth inhibitor. Missing out the spatial information would not permit to infer the mechanisms at work. Our findings suggest that this iterative data integration together with intermediate model sensitivity analysis at each model development stage, provide a promising strategy to infer predictive yet minimal (in the above sense) quantitative models of tumor growth, as prospectively of other tissue

  2. A Novel Multiparametric Drug-Scoring Method for High-Throughput Screening of 3D Multicellular Tumor Spheroids Using the Celigo Image Cytometer.

    Science.gov (United States)

    Cribbes, Scott; Kessel, Sarah; McMenemy, Scott; Qiu, Jean; Chan, Leo Li-Ying

    2017-06-01

    Three-dimensional (3D) tumor models have been increasingly used to investigate and characterize cancer drug compounds. The ability to perform high-throughput screening of 3D multicellular tumor spheroids (MCTS) can highly improve the efficiency and cost-effectiveness of discovering potential cancer drug candidates. Previously, the Celigo Image Cytometer has demonstrated a novel method for high-throughput screening of 3D multicellular tumor spheroids. In this work, we employed the Celigo Image Cytometer to examine the effects of 14 cancer drug compounds on 3D MCTS of the glioblastoma cell line U87MG in 384-well plates. Using parameters such as MCTS diameter and invasion area, growth and invasion were monitored for 9 and 3 d, respectively. Furthermore, fluorescent staining with calcein AM, propidium iodide, Hoechst 33342, and caspase 3/7 was performed at day 9 posttreatment to measure viability and apoptosis. Using the kinetic and endpoint data generated, we created a novel multiparametric drug-scoring system for 3D MCTS that can be used to identify and classify potential drug candidates earlier in the drug discovery process. Furthermore, the combination of quantitative and qualitative image data can be used to delineate differences between drugs that induce cytotoxic and cytostatic effects. The 3D MCTS-based multiparametric scoring method described here can provide an alternative screening method to better qualify tested drug compounds.

  3. Developing multi-cellular tumor spheroid model (MCTS) in the chitosan/collagen/alginate (CCA) fibrous scaffold for anticancer drug screening.

    Science.gov (United States)

    Wang, Jian-Zheng; Zhu, Yu-Xia; Ma, Hui-Chao; Chen, Si-Nan; Chao, Ji-Ye; Ruan, Wen-Ding; Wang, Duo; Du, Feng-guang; Meng, Yue-Zhong

    2016-05-01

    In this work, a 3D MCTS-CCA system was constructed by culturing multi-cellular tumor spheroid (MCTS) in the chitosan/collagen/alginate (CCA) fibrous scaffold for anticancer drug screening. The CCA scaffolds were fabricated by spray-spinning. The interactions between the components of the spray-spun fibers were evidenced by methods of Coomassie Blue stain, X-ray diffraction (XRD) and Fourier transform-infrared spectroscopy (FTIR). Co-culture indicated that MCF-7 cells showed a spatial growth pattern of multi-cellular tumor spheroid (MCTS) in the CCA fibrous scaffold with increased proliferation rate and drug-resistance to MMC, ADM and 5-Aza comparing with the 2D culture cells. Significant increases of total viable cells were found in 3D MCTS groups after drug administration by method of apoptotic analysis. Glucose-lactate analysis indicated that the metabolism of MCTS in CCA scaffold was closer to the tumor issue in vivo than the monolayer cells. In addition, MCTS showed the characteristic of epithelial mesenchymal transition (EMT) which is subverted by carcinoma cells to facilitate metastatic spread. These results demonstrated that MCTS in CCA scaffold possessed a more conservative phenotype of tumor than monolayer cells, and anticancer drug screening in 3D MCTS-CCA system might be superior to the 2D culture system. Copyright © 2016 Elsevier B.V. All rights reserved.

  4. Down-regulation of DNA mismatch repair proteins in human and murine tumor spheroids: implications for multicellular resistance to alkylating agents.

    Science.gov (United States)

    Francia, Giulio; Green, Shane K; Bocci, Guido; Man, Shan; Emmenegger, Urban; Ebos, John M L; Weinerman, Adina; Shaked, Yuval; Kerbel, Robert S

    2005-10-01

    Similar to other anticancer agents, intrinsic or acquired resistance to DNA-damaging chemotherapeutics is a major obstacle for cancer therapy. Current strategies aimed at overcoming this problem are mostly based on the premise that tumor cells acquire heritable genetic mutations that contribute to drug resistance. Here, we present evidence for an epigenetic, tumor cell adhesion-mediated, and reversible form of drug resistance that is associated with a reduction of DNA mismatch repair proteins PMS2 and/or MLH1 as well as other members of this DNA repair process. Growth of human breast cancer, human melanoma, and murine EMT-6 breast cancer cell lines as multicellular spheroids in vitro, which is associated with increased resistance to many chemotherapeutic drugs, including alkylating agents, is shown to lead to a reproducible down-regulation of PMS2, MLH1, or, in some cases, both as well as MHS6, MSH3, and MSH2. The observed down-regulation is in part reversible by treatment of tumor spheroids with the DNA-demethylating agent, 5-azacytidine. Thus, treatment of EMT-6 mouse mammary carcinoma spheroids with 5-azacytidine resulted in reduced and/or disrupted cell-cell adhesion, which in turn sensitized tumor spheroids to cisplatin-mediated killing in vitro. Our results suggest that antiadhesive agents might sensitize tumor spheroids to alkylating agents in part by reversing or preventing reduced DNA mismatch repair activity and that the chemosensitization properties of 5-azacytidine may conceivably reflect its role as a potential antiadhesive agent as well as reversal agent for MLH1 gene silencing in human tumors.

  5. Effect of combined treatment of x-rays and ACNU on rat glioma cells in monolayer and multicellular spheroids

    International Nuclear Information System (INIS)

    Sugiyama, Satoru; Mori, Teruaki; Suzuki, Jiro; Sasaki, Takehito

    1985-01-01

    Spheroids of rat glioma clone-6 cells having a central necrosis were used to determine the effect of combined treatment of x-rays and 1-(4-amino-2-methyl-5-pyrimidinyl)methyl-3-(2-chloroethyl)-3-nitrosourea hydrochloride (ACNU), where the optimum time intervals and doses in the combination were analyzed. The treatment with ACNU 2 to 6 hours prior to x-ray irradiation was most effective for cells in both monolayers and in spheroids. The dose survival curves with x-ray irradiation indicated that the hypoxic cell fraction in spheroids disappeared with a prior treatment by ACNU. The enhancement ratio in spheroids was thus larger for larger x-ray doses, and was always larger than that in monolayer cells. The survival curves versus concentration of ACNU indicated that the enhancement ratio in spheroids was more than 1.2 in all concentrations with the combined x-ray irradiation, and exceeded that in monolayer cells with a surviving fraction of less than 0.4. (author)

  6. Further evidence for the absence of a hypoxic fraction in the 9L rat tumour multicellular spheroid system

    International Nuclear Information System (INIS)

    Gutin, P.H.; Barcellos, M.H.; Shrieve, D.C.; Sano, Y.; Bernstein, M.; Deen, D.F.

    1982-01-01

    The 9L gliosarcoma is an N-methylnitrosourea-induced rat brain tumour that has served as a predictive model for the efficacy of various chemotherapeutic agents against human brain tumours. Because it is one of two known animal tumour models that has no hypoxic fraction, the 9L model is of questionable value for the study of the radiobiology of hypoxic cell sensitizers. Hypoxic 9L monolayer cells are sensitive to misonidazole, as shown by the abrupt decrease in survival after a 2-4 h radiation exposure. However, when 9L spheroids in the size ranges of 200-300, 300-400, 500-600 and 1027+-33μm were incubated in euoxic spinner culture for up to 96 h in 1.5 or 3.0 mM misonidazole, there was no effect on the survival of the dissociated cells over a dose range 0-20 Gy. It is concluded that, in view of the demonstrated sensitivity to misonidazole of hypoxic 9L cells in monolayer culture, this finding provides further evidence that there are no hypoxic cells even in large 9L spheroids with a histologically distinct zone of central necrosis. Moreover, 9L spheroids irradiated in the presence of 3.0 mM misonidazole showed no dose enhancement. (U.K.)

  7. In-situ birth of MSCs multicellular spheroids in poly(L-glutamic acid)/chitosan scaffold for hyaline-like cartilage regeneration.

    Science.gov (United States)

    Zhang, Kunxi; Yan, Shifeng; Li, Guifei; Cui, Lei; Yin, Jingbo

    2015-12-01

    The success of mesenchymal stem cells (MSCs) based articular cartilage tissue engineering is limited by the presence of fibrous tissue in generated cartilage, which is associated with the current scaffold strategy that promotes cellular adhesion and spreading. Here we design a non-fouling scaffold based on amide bonded poly(l-glutamic acid) (PLGA) and chitosan (CS) to drive adipose stem cells (ASCs) to aggregate to form multicellular spheroids with diameter of 80-110 μm in-situ. To illustrate the advantage of the present scaffolds, a cellular adhesive scaffold based on the same amide bonded PLGA and CS was created through a combination of air-drying and freeze-drying to limit the hydration effect while also achieving porous structure. Compared to ASCs spreading along the surface of pores within scaffold, the dense mass of aggregated ASCs in PLGA/CS scaffold exhibited enhanced chondrogenic differentiation capacity, as determined by up-regulated GAGs and COL II expression, and greatly decreased COL I deposition during in vitro chondrogenesis. Furthermore, after 12 weeks of implantation, neo-cartilages generated by ASCs adhered on scaffold significantly presented fibrous matrix which was characterized by high levels of COL I deposition. However, neo-cartilage at 12 weeks post-implantation generated by PLGA/CS scaffold carrying ASC spheroids possessed similar high level of GAGs and COL II and low level of COL I as that in normal cartilage. The in vitro and in vivo results indicated the present strategy could not only promote chondrogenesis of ASCs, but also facilitate hyaline-like cartilage regeneration with reduced fibrous tissue formation which may attenuate cartilage degradation in future long-term follow-up. Copyright © 2015 Elsevier Ltd. All rights reserved.

  8. Morphogenetic and neuronal characterization of human neuroblastoma multicellular spheroids cultured under undifferentiated and all-trans-retinoic acid-differentiated conditions

    Directory of Open Access Journals (Sweden)

    Gwon-Soo Jung

    2013-05-01

    Full Text Available In this study, we aimed to compare the morphogenetic andneuronal characteristics between monolayer cells andspheroids. For this purpose, we established spheroid formationby growing SH-SY5Y cells on the hydrophobic surfaces ofthermally-collapsed elastin-like polypeptide. After 4 days ofculture, the relative proliferation of the cells within spheroidswas approximately 92% of the values for monolayer cultures.As measured by quantitative assays for mRNA and proteinexpressions, the production of synaptophysin and neuronspecificenolase (NSE as well as the contents of cell adhesionmolecules (CAMs and extracellular matrix (ECM proteins aremuch higher in spheroids than in monolayer cells. Under theall-trans-retinoic acid (RA-induced differentiation condition,spheroids extended neurites and further up-regulated theexpression of synaptophysin, NSE, CAMs, and ECM proteins.Our data indicate that RA-differentiated SH-SY5Y neurospheroidsare functionally matured neuronal architectures. [BMBReports 2013; 46(5: 276-281

  9. A comparison of anti-tumor effects of high dose rate fractionated and low dose rate continuous irradiation in multicellular spheroids

    International Nuclear Information System (INIS)

    Kubota, Nobuo; Omura, Motoko; Matsubara, Sho.

    1997-01-01

    In a clinical experience, high dose rate (HDR) fractionated interstitial radiotherapy can be an alternative to traditional low dose rate (LDR) continuous interstitial radiotherapy for head and neck cancers. To investigate biological effect of HDR, compared to LDR, comparisons have been made using spheroids of human squamous carcinoma cells. Both LDR and HDR were delivered by 137 Cs at 37degC. Dose rate of LDR was 8 Gy/day and HDR irradiations of fraction size of 4, 5 or 6 Gy were applied twice a day with an interval time of more than 6 hr. We estimated HDR fractionated dose of 31 Gy with 4 Gy/fr to give the same biological effects of 38 Gy by continuous LDR for spheroids. The ratio of HDR/LDR doses to control 50% spheroids was 0.82. (author)

  10. Detection and analysis of human serum albumin nanoparticles within phagocytic cells at the resolution of individual live cell or single 3D multicellular spheroid

    Energy Technology Data Exchange (ETDEWEB)

    Afrimzon, Elena; Zurgil, Naomi; Sobolev, Maria; Shafran, Yana [Bar-Ilan University, The Biophysical Interdisciplinary Schottenstein Center for the Research and Technology of the Cellome (Israel); Langer, Klaus; Zlatev, Iavor [Westfälischen Wilhelms-Universität Münster, Institut für Pharmazeutische Technologie und Biopharmazie (Germany); Wronski, Robert; Windisch, Manfred [QPS Austria GmbH (Austria); Briesen, Hagen von [Fraunhofer Institute for Biomedical Engineering IBMT, Department of Cell Biology & Applied Virology (Germany); Schmidt, Reinhold [Medical University of Graz, Department of Neurology (Austria); Pietrzik, Claus [University Medical Center of the Johannes Gutenberg University of Mainz, Institute of Pathobiochemistry (Germany); Deutsch, Mordechai, E-mail: motti.jsc@gmail.com [Bar-Ilan University, The Biophysical Interdisciplinary Schottenstein Center for the Research and Technology of the Cellome (Israel)

    2015-12-15

    Since nanoparticles (NPs) have shown great potential in various biomedical applications, live cell response to NPs should be thoroughly explored prior to their in vivo use. In the current study, live cell array (LCA) methodology and unique cell-based assays were used to study the interaction of magnetite (HSA-Mag NP) loaded human serum albumin NPs with phagocytic cells. The LCA enabled cell culturing during HSA-Mag NP accumulation and monolayer or spheroid formation, concomitantly with on-line monitoring of NP internalization. These platforms were also utilized for imaging intercellular links between living cells preloaded with HSA-Mag NP in 2D and 3D resolution. HSA-Mag NP uptake by cells was quantified by imaging, and analyzed using time-resolved measurements. Image analysis of the individual cells in cell populations showed accumulation of HSA-Mag NP by promonocytes and glial cells in a dose- and time-dependent manner. High variability of NP accumulation in individual cells within cell populations, as well as cell subgroups, was evident in both cell types. Following 24 h interaction, uptake of HSA-Mag NP was about 10 times more efficient in glial cells than in activated promonocytes. The presented assays may facilitate detection and analysis of the amount of NPs within individual cells, as well as the rate of NP accumulation and processing in different subsets of living cells. Such data are crucial for estimating predicted drug dosage delivered by NPs, as well as to study possible mechanisms for NP interference with live cells.

  11. Heterogenic expression of stem cell markers in patient-derived glioblastoma spheroid cultures exposed to long-term hypoxia

    DEFF Research Database (Denmark)

    Rosenberg, Tine; Aaberg-Jessen, Charlotte; Petterson, Stine Asferg

    2018-01-01

    AIM: To investigate the time profile of hypoxia and stem cell markers in glioblastoma spheroids of known molecular subtype. MATERIALS & METHODS: Patient-derived glioblastoma spheroids were cultured up to 7 days in either 2% or 21% oxygen. Levels of proliferation (Ki-67), hypoxia (HIF-1α, CA9...

  12. Heterogenic expression of stem cell markers in patient-derived glioblastoma spheroid cultures exposed to long-term hypoxia

    DEFF Research Database (Denmark)

    Rosenberg, Tine; Aaberg-Jessen, Charlotte; Petterson, Stine Asferg

    2018-01-01

    AIM: To investigate the time profile of hypoxia and stem cell markers in glioblastoma spheroids of known molecular subtype. MATERIALS & METHODS: Patient-derived glioblastoma spheroids were cultured up to 7 days in either 2% or 21% oxygen. Levels of proliferation (Ki-67), hypoxia (HIF-1α, CA9...... and VEGF) and stem cell markers (CD133, nestin and musashi-1) were investigated by immunohistochemistry. RESULTS: Hypoxia markers as well as CD133 and partially nestin increased in long-term hypoxia. The proliferation rate and spheroid size were highest in normoxia. CONCLUSION: We found differences...... in hypoxia and stem cell marker profiles between the patient-derived glioblastoma cultures. This heterogeneity should be taken into consideration in development of future therapeutic strategies....

  13. Magnetically levitated mesenchymal stem cell spheroids cultured with a collagen gel maintain phenotype and quiescence

    Directory of Open Access Journals (Sweden)

    Natasha S Lewis

    2017-04-01

    Full Text Available Multicellular spheroids are an established system for three-dimensional cell culture. Spheroids are typically generated using hanging drop or non-adherent culture; however, an emerging technique is to use magnetic levitation. Herein, mesenchymal stem cell spheroids were generated using magnetic nanoparticles and subsequently cultured within a type I collagen gel, with a view towards developing a bone marrow niche environment. Cells were loaded with magnetic nanoparticles, and suspended beneath an external magnet, inducing self-assembly of multicellular spheroids. Cells in spheroids were viable and compared to corresponding monolayer controls, maintained stem cell phenotype and were quiescent. Interestingly, core spheroid necrosis was not observed, even with increasing spheroid size, in contrast to other commonly used spheroid systems. This mesenchymal stem cell spheroid culture presents a potential platform for modelling in vitro bone marrow stem cell niches, elucidating interactions between cells, as well as a useful model for drug delivery studies.

  14. The multicellularity genes of dictyostelid social amoebas

    Science.gov (United States)

    Glöckner, Gernot; Lawal, Hajara M.; Felder, Marius; Singh, Reema; Singer, Gail; Weijer, Cornelis J.; Schaap, Pauline

    2016-01-01

    The evolution of multicellularity enabled specialization of cells, but required novel signalling mechanisms for regulating cell differentiation. Early multicellular organisms are mostly extinct and the origins of these mechanisms are unknown. Here using comparative genome and transcriptome analysis across eight uni- and multicellular amoebozoan genomes, we find that 80% of proteins essential for the development of multicellular Dictyostelia are already present in their unicellular relatives. This set is enriched in cytosolic and nuclear proteins, and protein kinases. The remaining 20%, unique to Dictyostelia, mostly consists of extracellularly exposed and secreted proteins, with roles in sensing and recognition, while several genes for synthesis of signals that induce cell-type specialization were acquired by lateral gene transfer. Across Dictyostelia, changes in gene expression correspond more strongly with phenotypic innovation than changes in protein functional domains. We conclude that the transition to multicellularity required novel signals and sensors rather than novel signal processing mechanisms. PMID:27357338

  15. Autophagy Protects from Trastuzumab-Induced Cytotoxicity in HER2 Overexpressing Breast Tumor Spheroids.

    Directory of Open Access Journals (Sweden)

    Cristina E Rodríguez

    Full Text Available Multicellular tumor spheroids represent a 3D in vitro model that mimics solid tumor essential properties including assembly and development of extracellular matrix and nutrient, oxygen and proliferation gradients. In the present study, we analyze the impact of 3D spatial organization of HER2-overexpressing breast cancer cells on the response to Trastuzumab. We cultured human mammary adenocarcinoma cell lines as spheroids with the hanging drop method and we observed a gradient of proliferating, quiescent, hypoxic, apoptotic and autophagic cells towards the inner core. This 3D organization decreased Trastuzumab sensitivity of HER2 over-expressing cells compared to monolayer cell cultures. We did not observe apoptosis induced by Trastuzumab but found cell arrest in G0/G1 phase. Moreover, the treatment downregulated the basal apoptosis only found in tumor spheroids, by eliciting protective autophagy. We were able to increase sensitivity to Trastuzumab by autophagy inhibition, thus exposing the interaction between apoptosis and autophagy. We confirmed this result by developing a resistant cell line that was more sensitive to autophagy inhibition than the parental BT474 cells. In summary, the development of Trastuzumab resistance relies on the balance between death and survival mechanisms, characteristic of 3D cell organization. We propose the use of spheroids to further improve the understanding of Trastuzumab antitumor activity and overcome resistance.

  16. Digital microfluidics for automated hanging drop cell spheroid culture.

    Science.gov (United States)

    Aijian, Andrew P; Garrell, Robin L

    2015-06-01

    Cell spheroids are multicellular aggregates, grown in vitro, that mimic the three-dimensional morphology of physiological tissues. Although there are numerous benefits to using spheroids in cell-based assays, the adoption of spheroids in routine biomedical research has been limited, in part, by the tedious workflow associated with spheroid formation and analysis. Here we describe a digital microfluidic platform that has been developed to automate liquid-handling protocols for the formation, maintenance, and analysis of multicellular spheroids in hanging drop culture. We show that droplets of liquid can be added to and extracted from through-holes, or "wells," and fabricated in the bottom plate of a digital microfluidic device, enabling the formation and assaying of hanging drops. Using this digital microfluidic platform, spheroids of mouse mesenchymal stem cells were formed and maintained in situ for 72 h, exhibiting good viability (>90%) and size uniformity (% coefficient of variation <10% intraexperiment, <20% interexperiment). A proof-of-principle drug screen was performed on human colorectal adenocarcinoma spheroids to demonstrate the ability to recapitulate physiologically relevant phenomena such as insulin-induced drug resistance. With automatable and flexible liquid handling, and a wide range of in situ sample preparation and analysis capabilities, the digital microfluidic platform provides a viable tool for automating cell spheroid culture and analysis. © 2014 Society for Laboratory Automation and Screening.

  17. Cell cycle variation in x-ray survival for cells from spheroids measured by volume cell sorting

    International Nuclear Information System (INIS)

    Freyer, J.P.; Wilder, M.E.; Raju, M.R.

    1984-01-01

    Considerable work has been done studying the variation in cell survival as a function of cell cycle position for monolayers or single cells exposed to radiation. Little is known about the effects of multicellular growth on the relative radiation sensitivity of cells in different cell cycle stages. The authors have developed a new technique for measuring the response of cells, using volume cell sorting, which is rapid, non-toxic, and does not require cell synchronization. By combining this technique with selective spheroid dissociation,they have measured the age response of cells located at various depths in EMT6 and Colon 26 spheroids. Although cells in the inner region had mostly G1-phase DNA contents, 15-20% had S- and G2-phase DNA contents. Analysis of these cells using BrdU labeling and flow cytometric analysis with a monoclonal antibody to BrdU indicated that the inner region cells were not synthesizing DNA. Thus, the authors were able to measure the radiation response of cells arrested in G1, S and G2 cell cycle phases. Comparison of inner and outer spheroid regions, and monolayer cultures, indicates that it is improper to extrapolate age response data in standard culture conditions to the situation in spheroids

  18. Semiotic scaffolding of multicellularity

    DEFF Research Database (Denmark)

    Hoffmeyer, Jesper

    2015-01-01

    and animals this individuation process poses very different challenges in the three kingdoms of plants, fungi and animals, and the solutions found to these differences are discussed. In the same time as multicellularity ushered life into the epoch of mortality it logically also led to the appearance...

  19. Multicellular Features of Phytoplankton

    Directory of Open Access Journals (Sweden)

    Adi Abada

    2018-04-01

    Full Text Available Microscopic marine phytoplankton drift freely in the ocean, harvesting sunlight through photosynthesis. These unicellular microorganisms account for half of the primary productivity on Earth and play pivotal roles in the biogeochemistry of our planet (Field et al., 1998. The major groups of microalgae that comprise the phytoplankton community are coccolithophores, diatoms and dinoflagellates. In present oceans, phytoplankton individuals and populations are forced to rapidly adjust, as key chemical and physical parameters defining marine habitats are changing globally. Here we propose that microalgal populations often display the characteristics of a multicellular-like community rather than a random collection of individuals. Evolution of multicellularity entails a continuum of events starting from single cells that go through aggregation or clonal divisions (Brunet and King, 2017. Phytoplankton may be an intermediate state between single cells and aggregates of physically attached cells that communicate and co-operate; perhaps an evolutionary snapshot toward multicellularity. In this opinion article, we journey through several studies conducted in two key phytoplankton groups, coccolithophores and diatoms, to demonstrate how observations in these studies could be interpreted in a multicellular context.

  20. Games of multicellularity.

    Science.gov (United States)

    Kaveh, Kamran; Veller, Carl; Nowak, Martin A

    2016-08-21

    Evolutionary game dynamics are often studied in the context of different population structures. Here we propose a new population structure that is inspired by simple multicellular life forms. In our model, cells reproduce but can stay together after reproduction. They reach complexes of a certain size, n, before producing single cells again. The cells within a complex derive payoff from an evolutionary game by interacting with each other. The reproductive rate of cells is proportional to their payoff. We consider all two-strategy games. We study deterministic evolutionary dynamics with mutations, and derive exact conditions for selection to favor one strategy over another. Our main result has the same symmetry as the well-known sigma condition, which has been proven for stochastic game dynamics and weak selection. For a maximum complex size of n=2 our result holds for any intensity of selection. For n≥3 it holds for weak selection. As specific examples we study the prisoner's dilemma and hawk-dove games. Our model advances theoretical work on multicellularity by allowing for frequency-dependent interactions within groups. Copyright © 2016 Elsevier Ltd. All rights reserved.

  1. Comparative analysis of tumor spheroid generation techniques for differential in vitro drug toxicity

    Science.gov (United States)

    Raghavan, Shreya; Rowley, Katelyn R.; Mehta, Geeta

    2016-01-01

    Multicellular tumor spheroids are powerful in vitro models to perform preclinical chemosensitivity assays. We compare different methodologies to generate tumor spheroids in terms of resultant spheroid morphology, cellular arrangement and chemosensitivity. We used two cancer cell lines (MCF7 and OVCAR8) to generate spheroids using i) hanging drop array plates; ii) liquid overlay on ultra-low attachment plates; iii) liquid overlay on ultra-low attachment plates with rotating mixing (nutator plates). Analysis of spheroid morphometry indicated that cellular compaction was increased in spheroids generated on nutator and hanging drop array plates. Collagen staining also indicated higher compaction and remodeling in tumor spheroids on nutator and hanging drop arrays compared to conventional liquid overlay. Consequently, spheroids generated on nutator or hanging drop plates had increased chemoresistance to cisplatin treatment (20-60% viability) compared to spheroids on ultra low attachment plates (10-20% viability). Lastly, we used a mathematical model to demonstrate minimal changes in oxygen and cisplatin diffusion within experimentally generated spheroids. Our results demonstrate that in vitro methods of tumor spheroid generation result in varied cellular arrangement and chemosensitivity. PMID:26918944

  2. Formation of stable small cell number three-dimensional ovarian cancer spheroids using hanging drop arrays for preclinical drug sensitivity assays.

    Science.gov (United States)

    Raghavan, Shreya; Ward, Maria R; Rowley, Katelyn R; Wold, Rachel M; Takayama, Shuichi; Buckanovich, Ronald J; Mehta, Geeta

    2015-07-01

    Ovarian cancer grows and metastasizes from multicellular spheroidal aggregates within the ascites fluid. Multicellular tumor spheroids are therefore physiologically significant 3D in vitro models for ovarian cancer research. Conventional hanging drop cultures require high starting cell numbers, and are tedious for long-term maintenance. In this study, we generate stable, uniform multicellular spheroids using very small number of ovarian cancer cells in a novel 384 well hanging drop array platform. We used novel tumor spheroid platform and two ovarian cancer cell lines (A2780 and OVCAR3) to demonstrate the stable incorporation of as few as 10 cells into a single spheroid. Spheroids had uniform geometry, with projected areas (42.60×10(3)μm-475.22×10(3)μm(2) for A2780 spheroids and 37.24×10(3)μm(2)-281.01×10(3)μm(2) for OVCAR3 spheroids) that varied as a function of the initial cell seeding density. Phalloidin and nuclear stains indicated cells formed tightly packed spheroids with demarcated boundaries and cell-cell interaction within spheroids. Cells within spheroids demonstrated over 85% viability. 3D tumor spheroids demonstrated greater resistance (70-80% viability) to cisplatin chemotherapy compared to 2D cultures (30-50% viability). Ovarian cancer spheroids can be generated from limited cell numbers in high throughput 384 well plates with high viability. Spheroids demonstrate therapeutic resistance relative to cells in traditional 2D culture. Stable incorporation of low cell numbers is advantageous when translating this research to rare patient-derived cells. This system can be used to understand ovarian cancer spheroid biology, as well as carry out preclinical drug sensitivity assays. Copyright © 2015 Elsevier Inc. All rights reserved.

  3. Self-assembly of tissue spheroids on polymeric membranes.

    Science.gov (United States)

    Messina, Antonietta; Morelli, Sabrina; Forgacs, Gabor; Barbieri, Giuseppe; Drioli, Enrico; De Bartolo, Loredana

    2017-07-01

    In this study, multicellular tissue spheroids were fabricated on polymeric membranes in order to accelerate the fusion process and tissue formation. To this purpose, tissue spheroids composed of three different cell types, myoblasts, fibroblasts and neural cells, were formed and cultured on agarose and membranes of polycaprolactone (PCL) and chitosan (CHT). Membranes prepared by a phase-inversion technique display different physicochemical, mechanical and transport properties, which can affect the fusion process. The membranes accelerated the fusion process of a pair of spheroids with respect to the inert substrate. In this process, a critical role is played by the membrane properties, especially by their mechanical characteristics and oxygen and carbon dioxide mass transfer. The rate of fusion was quantified and found to be similar for fibroblast, myoblast and neural tissue spheroids on membranes, which completed the fusion within 3 days. These spheroids underwent faster fusion and maturation on PCL membrane than on agarose, the rate of fusion being proportional to the value of oxygen and carbon dioxide permeances and elastic characteristics. Consequently, tissue spheroids on the membranes expressed high biological activity in terms of oxygen uptake, making them more suitable as building blocks in the fabrication of tissues and organs. Copyright © 2015 John Wiley & Sons, Ltd. Copyright © 2015 John Wiley & Sons, Ltd.

  4. Development of Three-Dimensional Multicellular Tissue-Like Constructs for Mutational Analysis Using Macroporous Microcarriers

    Science.gov (United States)

    Jordan, Jacqueline A.; Fraga, Denise N.; Gonda, Steve R.

    2002-01-01

    A three-dimensional (3-D), tissue-like model was developed for the genotoxic assessment of space environment. In previous experiments, we found that culturing mammalian cells in a NASA-designed bioreactor, using Cytodex-3 beads as a scaffold, generated 3-D multicellular spheroids. In an effort to generate scaffold-free spheroids, we developed a new 3-D tissue-like model by coculturing fibroblast and epithelial cell in a NASA bioreactor using macroporous Cultispher-S(TradeMark) microcarriers. Big Blue(Registered Trademark) Rat 2(Lambda) fibroblasts, genetically engineered to contain multiple copies (>60 copies/cell) of the Lac I target gene, were cocultured with radio-sensitive human epithelial cells, H184F5. Over an 8-day period, samples were periodically examined by microscopy and histology to confirm cell attachment, growth, and viability. Immunohistochemistry and western analysis were used to evaluate the expression of specific cytoskeletal and adhesion proteins. Key cell culture parameters (glucose, pH, and lactate concentrations) were monitored daily. Controls were two-dimensional mono layers of fibroblast or epithelial cells cultured in T-flasks. Analysis of 3-D spheroids from the bioreactor suggests fibroblast cells attached to and completely covered the bead surface and inner channels by day 3 in the bioreactor. Treatment of the 3-day spheroids with dispase II dissolved the Cultisphers(TradeMark) and produced multicellular, bead-less constructs. Immunohistochemistry confirmed the presence of vi.mentin, cytokeratin and E-cadherin in treated spheroids. Examination of the dispase II treated spheroids with transmission electron microscopy (TEM) also showed the presence of desmosomes. These results suggest that the controlled enzymatic degradation of an artificial matrix in the low shear environment of the NASA-designed bioreactor can produce 3-D tissue-like spheroids. 2

  5. Modeling mechanical inhomogeneities in small populations of proliferating monolayers and spheroids.

    Science.gov (United States)

    Lejeune, Emma; Linder, Christian

    2018-06-01

    Understanding the mechanical behavior of multicellular monolayers and spheroids is fundamental to tissue culture, organism development, and the early stages of tumor growth. Proliferating cells in monolayers and spheroids experience mechanical forces as they grow and divide and local inhomogeneities in the mechanical microenvironment can cause individual cells within the multicellular system to grow and divide at different rates. This differential growth, combined with cell division and reorganization, leads to residual stress. Multiple different modeling approaches have been taken to understand and predict the residual stresses that arise in growing multicellular systems, particularly tumor spheroids. Here, we show that by using a mechanically robust agent-based model constructed with the peridynamic framework, we gain a better understanding of residual stresses in multicellular systems as they grow from a single cell. In particular, we focus on small populations of cells (1-100 s) where population behavior is highly stochastic and prior investigation has been limited. We compare the average strain energy density of cells in monolayers and spheroids using different growth and division rules and find that, on average, cells in spheroids have a higher strain energy density than cells in monolayers. We also find that cells in the interior of a growing spheroid are, on average, in compression. Finally, we demonstrate the importance of accounting for stochastic fluctuations in the mechanical environment, particularly when the cellular response to mechanical cues is nonlinear. The results presented here serve as a starting point for both further investigation with agent-based models, and for the incorporation of major findings from agent-based models into continuum scale models when explicit representation of individual cells is not computationally feasible.

  6. Encapsulation by Janus spheroids

    OpenAIRE

    Li, Wei; Liu, Ya; Brett, Genevieve; Gunton, James D.

    2011-01-01

    The micro/nano encapsulation technology has acquired considerable attention in the fields of drug delivery, biomaterial engineering, and materials science. Based on recent advances in chemical particle synthesis, we propose a primitive model of an encapsulation system produced by the self-assembly of Janus oblate spheroids, particles with oblate spheroidal bodies and two hemi-surfaces coded with dissimilar chemical properties. Using Monte Carlo simulation, we investigate the encapsulation sys...

  7. Fibroblast spheroids as a model to study sustained fibroblast quiescence and their crosstalk with tumor cells

    Energy Technology Data Exchange (ETDEWEB)

    Salmenperä, Pertteli, E-mail: pertteli.salmenpera@helsinki.fi [Department of Virology, Medicum, Faculty of Medicine, University of Helsinki, P.O. Box 21, FIN-00014 (Finland); Karhemo, Piia-Riitta [Research Programs Unit, Translational Cancer Biology, and Institute of Biomedicine, University of Helsinki, P.O. Box 63, FIN-00014 (Finland); Räsänen, Kati [Department of Virology, Medicum, Faculty of Medicine, University of Helsinki, P.O. Box 21, FIN-00014 (Finland); Laakkonen, Pirjo [Research Programs Unit, Translational Cancer Biology, and Institute of Biomedicine, University of Helsinki, P.O. Box 63, FIN-00014 (Finland); Vaheri, Antti [Department of Virology, Medicum, Faculty of Medicine, University of Helsinki, P.O. Box 21, FIN-00014 (Finland)

    2016-07-01

    Stromal fibroblasts have an important role in regulating tumor progression. Normal and quiescent fibroblasts have been shown to restrict and control cancer cell growth, while cancer-associated, i. e. activated fibroblasts have been shown to enhance proliferation and metastasis of cancer cells. In this study we describe generation of quiescent fibroblasts in multicellular spheroids and their effects on squamous cell carcinoma (SCC) growth in soft-agarose and xenograft models. Quiescent phenotype of fibroblasts was determined by global down-regulation of expression of genes related to cell cycle and increased expression of p27. Interestingly, microarray analysis showed that fibroblast quiescence was associated with similar secretory phenotype as seen in senescence and they expressed senescence-associated-β-galactosidase. Quiescent fibroblasts spheroids also restricted the growth of RT3 SCC cells both in soft-agarose and xenograft models unlike proliferating fibroblasts. Restricted tumor growth was associated with marginally increased tumor cell senescence and cellular differentiation, showed with senescence-associated-β-galactosidase and cytokeratin 7 staining. Our results show that the fibroblasts spheroids can be used as a model to study cellular quiescence and their effects on cancer cell progression. - Highlights: • Fibroblasts acquire a sustained quiescence when grown as multicellular spheroids. • This quiescence is associated with drastic change in gene expression. • Fibroblasts spheroids secrete various inflammation-linked cytokines and chemokines. • Fibroblasts spheroids reduced growth of RT3 SCC cells in xenograft model.

  8. Multiscale image analysis reveals structural heterogeneity of the cell microenvironment in homotypic spheroids.

    Science.gov (United States)

    Schmitz, Alexander; Fischer, Sabine C; Mattheyer, Christian; Pampaloni, Francesco; Stelzer, Ernst H K

    2017-03-03

    Three-dimensional multicellular aggregates such as spheroids provide reliable in vitro substitutes for tissues. Quantitative characterization of spheroids at the cellular level is fundamental. We present the first pipeline that provides three-dimensional, high-quality images of intact spheroids at cellular resolution and a comprehensive image analysis that completes traditional image segmentation by algorithms from other fields. The pipeline combines light sheet-based fluorescence microscopy of optically cleared spheroids with automated nuclei segmentation (F score: 0.88) and concepts from graph analysis and computational topology. Incorporating cell graphs and alpha shapes provided more than 30 features of individual nuclei, the cellular neighborhood and the spheroid morphology. The application of our pipeline to a set of breast carcinoma spheroids revealed two concentric layers of different cell density for more than 30,000 cells. The thickness of the outer cell layer depends on a spheroid's size and varies between 50% and 75% of its radius. In differently-sized spheroids, we detected patches of different cell densities ranging from 5 × 10 5 to 1 × 10 6  cells/mm 3 . Since cell density affects cell behavior in tissues, structural heterogeneities need to be incorporated into existing models. Our image analysis pipeline provides a multiscale approach to obtain the relevant data for a system-level understanding of tissue architecture.

  9. How 5000 independent rowers coordinate their strokes in order to row into the sunlight: Phototaxis in the multicellular green alga Volvox

    Directory of Open Access Journals (Sweden)

    Matsunaga Shigeru

    2010-07-01

    Full Text Available Abstract Background The evolution of multicellular motile organisms from unicellular ancestors required the utilization of previously evolved tactic behavior in a multicellular context. Volvocine green algae are uniquely suited for studying tactic responses during the transition to multicellularity because they range in complexity from unicellular to multicellular genera. Phototactic responses are essential for these flagellates because they need to orientate themselves to receive sufficient light for photosynthesis, but how does a multicellular organism accomplish phototaxis without any known direct communication among cells? Several aspects of the photoresponse have previously been analyzed in volvocine algae, particularly in the unicellular alga Chlamydomonas. Results In this study, the phototactic behavior in the spheroidal, multicellular volvocine green alga Volvox rousseletii (Volvocales, Chlorophyta was analyzed. In response to light stimuli, not only did the flagella waveform and beat frequency change, but the effective stroke was reversed. Moreover, there was a photoresponse gradient from the anterior to the posterior pole of the spheroid, and only cells of the anterior hemisphere showed an effective response. The latter caused a reverse of the fluid flow that was confined to the anterior hemisphere. The responsiveness to light is consistent with an anterior-to-posterior size gradient of eyespots. At the posterior pole, the eyespots are tiny or absent, making the corresponding cells appear to be blind. Pulsed light stimulation of an immobilized spheroid was used to simulate the light fluctuation experienced by a rotating spheroid during phototaxis. The results demonstrated that in free-swimming spheroids, only those cells of the anterior hemisphere that face toward the light source reverse the beating direction in the presence of illumination; this behavior results in phototactic turning. Moreover, positive phototaxis is facilitated by

  10. On the evolution of bacterial multicellularity.

    Science.gov (United States)

    Lyons, Nicholas A; Kolter, Roberto

    2015-04-01

    Multicellularity is one of the most prevalent evolutionary innovations and nowhere is this more apparent than in the bacterial world, which contains many examples of multicellular organisms in a surprising array of forms. Due to their experimental accessibility and the large and diverse genomic data available, bacteria enable us to probe fundamental aspects of the origins of multicellularity. Here we discuss examples of multicellular behaviors in bacteria, the selective pressures that may have led to their evolution, possible origins and intermediate stages, and whether the ubiquity of apparently convergent multicellular forms argues for its inevitability. Copyright © 2015 Elsevier Ltd. All rights reserved.

  11. Interferon-β lipofection I. Increased efficacy of chemotherapeutic drugs on human tumor cells derived monolayers and spheroids.

    Science.gov (United States)

    Villaverde, M S; Gil-Cardeza, M L; Glikin, G C; Finocchiaro, L M E

    2012-07-01

    We evaluated the effect of hIFNβ gene transfer alone or in combination with different antineoplastic drugs commonly used in cancer treatment. Five human tumor-derived cell lines were cultured as monolayers and spheroids. Four cell lines (Ewing sarcomas EW7 and COH, melanoma M8 and mammary carcinoma MCF-7) were sensitive to hIFNβ gene lipofection. Although this effect appeared in both culture configurations, spheroids showed a relative multicellular resistance (insensitive colon carcinoma HT-29 excluded). EW7 and M8 hIFNβ-expressing cells were exposed to different concentrations of bleomycin, bortezomib, carboplatin, doxorubicin, etoposide, methotrexate, paclitaxel and vincristine in both configuration models. In chemotherapy-sensitive EW7 monolayers, the combination of hIFNβ gene and antineoplastic drugs displayed only additive or counteractive (methotrexate) effects, suggesting that cytotoxic mechanisms triggered by hIFNβ gene lipofection could be saturating the signaling pathways. Conversely, in chemotherapy-resistant EW7 spheroids or M8 cells, the combination of hIFNβ with drugs that mainly operate at the genotoxic level (doxorubicin, methotrexate and paclitaxel) presented only additive effects. However, drugs that also increase pro-oxidant species can complement the antitumor efficacy of the hIFNβ gene and clearly caused potentiated effects (bleomycin, bortezomib, carboplatin, etoposide and vincristine). The great bystander effect induced by hIFNβ gene lipofection could be among the main causes of its effectiveness, because only 1 or 2% of EW7 or M8 hIFNβ-expressing cells killed more than 60 or 80% of cell population, respectively.

  12. Directed self-assembly of large scaffold-free multi-cellular honeycomb structures

    International Nuclear Information System (INIS)

    Tejavibulya, Nalin; Youssef, Jacquelyn; Bao, Brian; Ferruccio, Toni-Marie; Morgan, Jeffrey R

    2011-01-01

    A significant challenge to the field of biofabrication is the rapid construction of large three-dimensional (3D) living tissues and organs. Multi-cellular spheroids have been used as building blocks. In this paper, we create large multi-cellular honeycomb building blocks using directed self-assembly, whereby cell-to-cell adhesion, in the context of the shape and obstacles of a micro-mold, drives the formation of a 3D structure. Computer-aided design, rapid prototyping and replica molding were used to fabricate honeycomb-shaped micro-molds. Nonadhesive hydrogels cast from these micro-molds were equilibrated in the cell culture medium and seeded with two types of mammalian cells. The cells settled into the honeycomb recess were unable to attach to the nonadhesive hydrogel and so cell-to-cell adhesion drove the self-assembly of a large multi-cellular honeycomb within 24 h. Distinct morphological changes occurred to the honeycomb and its cells indicating the presence of significant cell-mediated tension. Unlike the spheroid, whose size is constrained by a critical diffusion distance needed to maintain cell viability, the overall size of the honeycomb is not limited. The rapid production of the honeycomb building unit, with its multiple rings of high-density cells and open lumen spaces, offers interesting new possibilities for biofabrication strategies.

  13. NADPH oxidase: an enzyme for multicellularity?

    Science.gov (United States)

    Lalucque, Hervé; Silar, Philippe

    2003-01-01

    Multicellularity has evolved several times during the evolution of eukaryotes. One evolutionary pressure that permits multicellularity relates to the division of work, where one group of cells functions as nutrient providers and the other in specialized roles such as defence or reproduction. This requires signalling systems to ensure harmonious development of multicellular structures. Here, we show that NADPH oxidases are specifically present in organisms that differentiate multicellular structures during their life cycle and are absent from unicellular life forms. The biochemical properties of these enzymes make them ideal candidates for a role in intercellular signalling.

  14. An mDia2/ROCK signaling axis regulates invasive egress from epithelial ovarian cancer spheroids.

    Science.gov (United States)

    Pettee, Krista M; Dvorak, Kaitlyn M; Nestor-Kalinoski, Andrea L; Eisenmann, Kathryn M

    2014-01-01

    Multi-cellular spheroids are enriched in ascites of epithelial ovarian cancer (OvCa) patients. They represent an invasive and chemoresistant cellular population fundamental to metastatic dissemination. The molecular mechanisms triggering single cell invasive egress from spheroids remain enigmatic. mDia formins are Rho GTPase effectors that are key regulators of F-actin cytoskeletal dynamics. We hypothesized that mDia2-driven F-actin dynamics promote single cell invasive transitions in clinically relevant three-dimensional (3D) OvCa spheroids. The current study is a dissection of the contribution of the F-actin assembly factor mDia2 formin in invasive transitions and using a clinically relevant ovarian cancer spheroid model. We show that RhoA-directed mDia2 activity is required for tight spheroid organization, and enrichment of mDia2 in the invasive cellular protrusions of collagen-embedded OVCA429 spheroids. Depleting mDia2 in ES-2 spheroids enhanced invasive dissemination of single amoeboid-shaped cells. This contrasts with spheroids treated with control siRNA, where a mesenchymal invasion program predominated. Inhibition of another RhoA effector, ROCK, had no impact on ES-2 spheroid formation but dramatically inhibited spheroid invasion through induction of a highly elongated morphology. Concurrent inhibition of ROCK and mDia2 blocked single cell invasion from ES-2 spheroids more effectively than inhibition of either protein alone, indicating that invasive egress of amoeboid cells from mDia2-depleted spheroids is ROCK-dependent. Our findings indicate that multiple GTPase effectors must be suppressed in order to fully block invasive egress from ovarian cancer spheroids. Furthermore, tightly regulated interplay between ROCK and mDia2 signaling pathways dictates the invasive capacities and the type of invasion program utilized by motile spheroid-derived ovarian cancer cells. As loss of the gene encoding mDia2, DRF3, has been linked to cancer progression and

  15. An mDia2/ROCK signaling axis regulates invasive egress from epithelial ovarian cancer spheroids.

    Directory of Open Access Journals (Sweden)

    Krista M Pettee

    Full Text Available Multi-cellular spheroids are enriched in ascites of epithelial ovarian cancer (OvCa patients. They represent an invasive and chemoresistant cellular population fundamental to metastatic dissemination. The molecular mechanisms triggering single cell invasive egress from spheroids remain enigmatic. mDia formins are Rho GTPase effectors that are key regulators of F-actin cytoskeletal dynamics. We hypothesized that mDia2-driven F-actin dynamics promote single cell invasive transitions in clinically relevant three-dimensional (3D OvCa spheroids. The current study is a dissection of the contribution of the F-actin assembly factor mDia2 formin in invasive transitions and using a clinically relevant ovarian cancer spheroid model. We show that RhoA-directed mDia2 activity is required for tight spheroid organization, and enrichment of mDia2 in the invasive cellular protrusions of collagen-embedded OVCA429 spheroids. Depleting mDia2 in ES-2 spheroids enhanced invasive dissemination of single amoeboid-shaped cells. This contrasts with spheroids treated with control siRNA, where a mesenchymal invasion program predominated. Inhibition of another RhoA effector, ROCK, had no impact on ES-2 spheroid formation but dramatically inhibited spheroid invasion through induction of a highly elongated morphology. Concurrent inhibition of ROCK and mDia2 blocked single cell invasion from ES-2 spheroids more effectively than inhibition of either protein alone, indicating that invasive egress of amoeboid cells from mDia2-depleted spheroids is ROCK-dependent. Our findings indicate that multiple GTPase effectors must be suppressed in order to fully block invasive egress from ovarian cancer spheroids. Furthermore, tightly regulated interplay between ROCK and mDia2 signaling pathways dictates the invasive capacities and the type of invasion program utilized by motile spheroid-derived ovarian cancer cells. As loss of the gene encoding mDia2, DRF3, has been linked to cancer

  16. Imaging Herpes Simplex Virus Type 1 Amplicon Vector–Mediated Gene Expression in Human Glioma Spheroids

    OpenAIRE

    Christine Kaestle; Alexandra Winkeler; Raphaela Richter; Heinrich Sauer; Jürgen Hescheler; Cornel Fraefel; Maria Wartenberg; Andreas H. Jacobs

    2011-01-01

    Vectors derived from herpes simplex virus type 1 (HSV-1) have great potential for transducing therapeutic genes into the central nervous system; however, inefficient distribution of vector particles in vivo may limit their therapeutic potential in patients with gliomas. This study was performed to investigate the extent of HSV-1 amplicon vector–mediated gene expression in a three-dimensional glioma model of multicellular spheroids by imaging highly infectious HSV-1 virions expressing green fl...

  17. Predictive modeling of multicellular structure formation by using Cellular Particle Dynamics simulations

    Science.gov (United States)

    McCune, Matthew; Shafiee, Ashkan; Forgacs, Gabor; Kosztin, Ioan

    2014-03-01

    Cellular Particle Dynamics (CPD) is an effective computational method for describing and predicting the time evolution of biomechanical relaxation processes of multicellular systems. A typical example is the fusion of spheroidal bioink particles during post bioprinting structure formation. In CPD cells are modeled as an ensemble of cellular particles (CPs) that interact via short-range contact interactions, characterized by an attractive (adhesive interaction) and a repulsive (excluded volume interaction) component. The time evolution of the spatial conformation of the multicellular system is determined by following the trajectories of all CPs through integration of their equations of motion. CPD was successfully applied to describe and predict the fusion of 3D tissue construct involving identical spherical aggregates. Here, we demonstrate that CPD can also predict tissue formation involving uneven spherical aggregates whose volumes decrease during the fusion process. Work supported by NSF [PHY-0957914]. Computer time provided by the University of Missouri Bioinformatics Consortium.

  18. Cell survival in spheroids irradiated with heavy-ion beams

    International Nuclear Information System (INIS)

    Rodriguez, A.; Alpen, E.L.

    1981-01-01

    Biological investigations with accelerated heavy ions have been carried out regularly at the Lawrence Berkeley Laboratory Bevalac for the past four years. Most of the cellular investigations have been conducted on cell monolayer and suspension culture systems. The studies to date suggest that heavy charged particle beams may offer some radiotherapeutic advantages over conventional radiotherapy sources. The advantages are thought to lie primarily in an increased relative biological effectiveness (RBE), a decrease in the oxygen enhancement ratio (OER), and better tissue distribution dose. Experiments reported here were conducted with 400 MeV/amu carbon ions and 425 MeV/amu neon ions, using a rat brain gliosarcoma cell line grown as multicellular spheroids. Studies have been carried out with x-rays and high-energy carbon and neon ion beams. These studies evaluate high-LET (linear energy transfer) cell survival in terms of RBE and the possible contributions of intercellular communication. Comparisons were made of the post-irradiation survival characteristics for cells irradiated as multicellular spheroids (approximately 100 μm and 300 μm diameters) and for cells irradiated in suspension. These comparisons were made between 225-kVp x-rays, 400 MeV/amu carbon ions, and 425 MeV/amu neon ions

  19. Spatio-temporal cell dynamics in tumour spheroid irradiation

    International Nuclear Information System (INIS)

    Kempf, H.; Bleicher, M.; Meyer-Hermann, M.; Kempf, H.; Bleicher, M.; Kempf, H.; Meyer-Hermann, M.

    2010-01-01

    Multicellular tumour spheroids are realistic in vitro systems in radiation research that integrate cell-cell interaction and cell cycle control by factors in the medium. The dynamic reaction inside a tumour spheroid triggered by radiation is not well understood. Of special interest is the amount of cell cycle synchronization which could be triggered by irradiation, since this would allow follow-up irradiations to exploit the increased sensitivity of certain cell cycle phases. In order to investigate these questions we need to support irradiation experiments with mathematical models. In this article a new model is introduced combining the dynamics of tumour growth and irradiation treatments. The tumour spheroid growth is modelled using an agent-based Delaunay/Voronoi hybrid model in which the cells are represented by weighted dynamic vertices. Cell properties like full cell cycle dynamics are included. In order to be able to distinguish between different cell reactions in response to irradiation quality we introduce a probabilistic model for damage dynamics. The overall cell survival from this model is in agreement with predictions from the linear-quadratic model. Our model can describe the growth of avascular tumour spheroids in agreement to experimental results. Using the probabilistic model for irradiation damage dynamics the classic 'four Rs' of radiotherapy can be studied in silico. We found a pronounced reactivation of the tumour spheroid in response to irradiation. A majority of the surviving cells is synchronized in their cell cycle progression after irradiation. The cell synchronization could be actively triggered and should be exploited in an advanced fractionation scheme. Thus it has been demonstrated that our model could be used to understand the dynamics of tumour growth after irradiation and to propose optimized fractionation schemes in cooperation with experimental investigations. (authors)

  20. The origin of multicellularity in cyanobacteria

    Science.gov (United States)

    2011-01-01

    Background Cyanobacteria are one of the oldest and morphologically most diverse prokaryotic phyla on our planet. The early development of an oxygen-containing atmosphere approximately 2.45 - 2.22 billion years ago is attributed to the photosynthetic activity of cyanobacteria. Furthermore, they are one of the few prokaryotic phyla where multicellularity has evolved. Understanding when and how multicellularity evolved in these ancient organisms would provide fundamental information on the early history of life and further our knowledge of complex life forms. Results We conducted and compared phylogenetic analyses of 16S rDNA sequences from a large sample of taxa representing the morphological and genetic diversity of cyanobacteria. We reconstructed ancestral character states on 10,000 phylogenetic trees. The results suggest that the majority of extant cyanobacteria descend from multicellular ancestors. Reversals to unicellularity occurred at least 5 times. Multicellularity was established again at least once within a single-celled clade. Comparison to the fossil record supports an early origin of multicellularity, possibly as early as the "Great Oxygenation Event" that occurred 2.45 - 2.22 billion years ago. Conclusions The results indicate that a multicellular morphotype evolved early in the cyanobacterial lineage and was regained at least once after a previous loss. Most of the morphological diversity exhibited in cyanobacteria today —including the majority of single-celled species— arose from ancient multicellular lineages. Multicellularity could have conferred a considerable advantage for exploring new niches and hence facilitated the diversification of new lineages. PMID:21320320

  1. Imaging Herpes Simplex Virus Type 1 Amplicon Vector–Mediated Gene Expression in Human Glioma Spheroids

    Directory of Open Access Journals (Sweden)

    Christine Kaestle

    2011-05-01

    Full Text Available Vectors derived from herpes simplex virus type 1 (HSV-1 have great potential for transducing therapeutic genes into the central nervous system; however, inefficient distribution of vector particles in vivo may limit their therapeutic potential in patients with gliomas. This study was performed to investigate the extent of HSV-1 amplicon vector–mediated gene expression in a three-dimensional glioma model of multicellular spheroids by imaging highly infectious HSV-1 virions expressing green fluorescent protein (HSV-GFP. After infection or microscopy-guided vector injection of glioma spheroids at various spheroid sizes, injection pressures and injection times, the extent of HSV-1 vector–mediated gene expression was investigated via laser scanning microscopy. Infection of spheroids with HSV-GFP demonstrated a maximal depth of vector-mediated GFP expression at 70 to 80 μm. A > 80% transduction efficiency was reached only in small spheroids with a diameter of 90%. The results demonstrated that vector-mediated gene expression in glioma spheroids was strongly dependent on the mode of vector application—injection pressure and injection time being the most important parameters. The assessment of these vector application parameters in tissue models will contribute to the development of safe and efficient gene therapy protocols for clinical application.

  2. Three-dimensional spheroid culture targeting versatile tissue bioassays using a PDMS-based hanging drop array.

    Science.gov (United States)

    Kuo, Ching-Te; Wang, Jong-Yueh; Lin, Yu-Fen; Wo, Andrew M; Chen, Benjamin P C; Lee, Hsinyu

    2017-06-29

    Biomaterial-based tissue culture platforms have emerged as useful tools to mimic in vivo physiological microenvironments in experimental cell biology and clinical studies. We describe herein a three-dimensional (3D) tissue culture platform using a polydimethylsiloxane (PDMS)-based hanging drop array (PDMS-HDA) methodology. Multicellular spheroids can be achieved within 24 h and further boosted by incorporating collagen fibrils in PDMS-HDA. In addition, the spheroids generated from different human tumor cells exhibited distinct sensitivities toward drug chemotherapeutic agents and radiation as compared with two-dimensional (2D) cultures that often lack in vivo-like biological insights. We also demonstrated that multicellular spheroids may enable key hallmarks of tissue-based bioassays, including drug screening, tumor dissemination, cell co-culture, and tumor invasion. Taken together, these results offer new opportunities not only to achieve the active control of 3D multicellular spheroids on demand, but also to establish a rapid and cost-effective platform to study anti-cancer therapeutics and tumor microenvironments.

  3. Multicellularity makes somatic differentiation evolutionarily stable

    Science.gov (United States)

    Wahl, Mary E.; Murray, Andrew W.

    2016-01-01

    Many multicellular organisms produce two cell lineages: germ cells, whose descendants produce the next generation, and somatic cells, which support, protect, and disperse the germ cells. This germ-soma demarcation has evolved independently in dozens of multicellular taxa but is absent in unicellular species. A common explanation holds that in these organisms, inefficient intercellular nutrient exchange compels the fitness cost of producing nonreproductive somatic cells to outweigh any potential benefits. We propose instead that the absence of unicellular, soma-producing populations reflects their susceptibility to invasion by nondifferentiating mutants that ultimately eradicate the soma-producing lineage. We argue that multicellularity can prevent the victory of such mutants by giving germ cells preferential access to the benefits conferred by somatic cells. The absence of natural unicellular, soma-producing species previously prevented these hypotheses from being directly tested in vivo: to overcome this obstacle, we engineered strains of the budding yeast Saccharomyces cerevisiae that differ only in the presence or absence of multicellularity and somatic differentiation, permitting direct comparisons between organisms with different lifestyles. Our strains implement the essential features of irreversible conversion from germ line to soma, reproductive division of labor, and clonal multicellularity while maintaining sufficient generality to permit broad extension of our conclusions. Our somatic cells can provide fitness benefits that exceed the reproductive costs of their production, even in unicellular strains. We find that nondifferentiating mutants overtake unicellular populations but are outcompeted by multicellular, soma-producing strains, suggesting that multicellularity confers evolutionary stability to somatic differentiation. PMID:27402737

  4. Evaluation of anti-HER2 scFv-conjugated PLGA–PEG nanoparticles on 3D tumor spheroids of BT474 and HCT116 cancer cells

    International Nuclear Information System (INIS)

    Le, Thi Thuy Duong; Pham, Thu Hong; Ngo, Thi Hong Giang; Le, Quang Huan; Nguyen, Trong Nghia; Hoang, Thi My Nhung

    2016-01-01

    Three-dimensional culture cells (spheroids) are one of the multicellular culture models that can be applied to anticancer chemotherapeutic development. Multicellular spheroids more closely mimic in vivo tumor-like patterns of physiologic environment and morphology. In previous research, we designed docetaxel-loaded pegylated poly(D, L-lactide-co-glycolide) nanoparticles conjugated with anti-HER2 single chain antibodies (scFv–Doc–PLGA–PEG) and evaluated them in 2D cell culture. In this study, we continuously evaluate the cellular uptake and cytotoxic effect of scFv–Doc–PLGA–PEG on a 3D tumor spheroid model of BT474 (HER2-overexpressing) and HCT116 (HER2-underexpressing) cancer cells. The results showed that the nanoparticle formulation conjugated with scFv had a significant internalization effect on the spheroids of HER2-overexpressing cancer cells as compared to the spheroids of HER2-underexpressing cancer cells. Therefore, cytotoxic effects of targeted nanoparticles decreased the size and increased necrotic score of HER2-overexpressing tumor spheroids. Thus, these scFv–Doc–PLGA–PEG nanoparticles have potential for active targeting for HER2-overexpressing cancer therapy. In addition, BT474 and HCT116 spheroids can be used as a tumor model for evaluation of targeting therapies. (paper)

  5. Bacterial Ventures into Multicellularity: Collectivism through Individuality.

    Directory of Open Access Journals (Sweden)

    Simon van Vliet

    2015-06-01

    Full Text Available Multicellular eukaryotes can perform functions that exceed the possibilities of an individual cell. These functions emerge through interactions between differentiated cells that are precisely arranged in space. Bacteria also form multicellular collectives that consist of differentiated but genetically identical cells. How does the functionality of these collectives depend on the spatial arrangement of the differentiated bacteria? In a previous issue of PLOS Biology, van Gestel and colleagues reported an elegant example of how the spatial arrangement of differentiated cells gives rise to collective behavior in Bacillus subtilus colonies, further demonstrating the similarity of bacterial collectives to higher multicellular organisms.

  6. Halo carbon stars associated with dwarf spheroidal galaxies

    Energy Technology Data Exchange (ETDEWEB)

    Van Den Bergh, S.; Lafontaine, A.

    1984-11-01

    Star counts have been performed for rings centered on the carbon star at 1 69 degrees, b + 55 degrees at a distance of 60 kpc. The counts were performed in order to determine whether halo carbon stars might be situated in dwarf spheroidal galaxies which are too star-poor to have been recognized as galaxies. The counts were made on a IIIa-J plate baked in forming gas that was exposed for 40 minutes through a 2C filter with the Palomar 1.2-m Schmidt telescope. It is shown that the carbon star is not situated in a dwarf spheroidal galaxy brighter than M(V) 5.7.

  7. Isotropic oscillator: spheroidal wave functions

    International Nuclear Information System (INIS)

    Mardoyan, L.G.; Pogosyan, G.S.; Ter-Antonyan, V.M.; Sisakyan, A.N.

    1985-01-01

    Solutions of the Schroedinger equation are found for an isotropic oscillator (10) in prolate and oblate spheroidal coordinates. It is shown that the obtained solutions turn into spherical and cylindrical bases of the isotropic oscillator at R→0 and R→ infinity (R is the dimensional parameter entering into the definition of prolate and oblate spheroidal coordinates). The explicit form is given for both prolate and oblate basis of the isotropic oscillator for the lowest quantum states

  8. The Andromeda Dwarf Spheroidal Galaxies

    OpenAIRE

    Armandroff, Taft E.; Da Costa, Gary S.

    1998-01-01

    Our current knowledge of M31's dwarf spheroidal companions is reviewed. Two topics of recent interest constitute the bulk of this review. First, color-magnitude diagrams reaching below the horizontal branch have been constructed for two M31 dwarf spheroidals based on images from HST/WFPC2. The horizontal branches are predominantly red in both galaxies, redder than expected for their metallicity based on Galactic globular clusters. Thus, the second parameter effect is seen in the M31 halo. Sec...

  9. Engineering emergent multicellular behavior through synthetic adhesion

    Science.gov (United States)

    Glass, David; Riedel-Kruse, Ingmar

    In over a decade, synthetic biology has developed increasingly robust gene networks within single cells, but constructed very few systems that demonstrate multicellular spatio-temporal dynamics. We are filling this gap in synthetic biology's toolbox by developing an E. coli self-assembly platform based on modular cell-cell adhesion. We developed a system in which adhesive selectivity is provided by a library of outer membrane-displayed peptides with intra-library specificities, while affinity is provided by consistent expression across the entire library. We further provide a biophysical model to help understand the parameter regimes in which this tool can be used to self-assemble into cellular clusters, filaments, or meshes. The combined platform will enable future development of synthetic multicellular systems for use in consortia-based metabolic engineering, in living materials, and in controlled study of minimal multicellular systems. Stanford Bio-X Bowes Fellowship.

  10. Long-term culture of organotypic multicellular glioma spheroids: a good culture model for studying gliomas

    NARCIS (Netherlands)

    Kaaijk, P.; Troost, D.; Das, P. K.; Leenstra, S.; Bosch, D. A.

    1995-01-01

    Gliomas, as well as other solid tumours, contain tumour stroma composed of connective tissue, macrophages, capillaries and other non-cellular constituents. Therefore, a homogeneous culture of tumour cells alone, as is often used as a culture model for gliomas, is not ideal to study all aspects of

  11. Targeted radiotherapy of multicell neuroblastoma spheroids with high specific activity [125I]meta-iodobenzylguanidine

    International Nuclear Information System (INIS)

    Roa, Wilson H.Y.; Miller, Gerald G.; McEwan, Alexander J.B.; McQuarrie, Steve A.; Tse, Jeanie; Wu, Jonn; Wiebe, Leonard I.

    1998-01-01

    Purpose: Iodine-125 induces cell death by a mechanism similar to that of high linear energy transfer (high-LET) radiation. This study investigates the cytotoxicity of high-specific-activity [ 125 I]meta-iodobenzylguanidine ( 125 I-mIBG) in human SK-N-MC neuroblastoma cells grown as three-dimensional multicellular spheroids. Materials and Methods: Spheroids were incubated with high-specific-activity 125 I-mIBG (6 mCi/μg, 1000 times that of the conventional specific activity used for autoradiography). Cytotoxicity was assessed by fluorescence viability markers and confocal microscopy for intact spheroids, fluorescence-activated cell sorting and clonogenic assay, and clonogenic assays for dispersed whole spheroids. Distribution of radioactive mIBG was determined by quantitative light-microscope autoradiography of spheroid cryostat sections. Dose estimation was based on temporal knowledge of the retained radioactivity inside spheroids, and of the radiolabel's emission characteristics. Findings were compared with those of spheroids treated under the same conditions with 131 I-mIBG, cold mIBG, and free iodine-125. Results: 125 I-mIBG exerted significant cell killing. Complete spheroids were eradicated when they were treated with 500 μCi of 125 I-mIBG, while those treated with 500 μCi or 1000 μCi of 131 I-mIBG were not. The observed difference in cytotoxicity between treatments with 125 I- and 131 I-mIBG could not be accounted for by the absorbed dose of spheroid alone. The peripheral, proliferating cell layer of the spheroids remained viable at the moderate radioactivity of 100 μCi for both isotopes. Cytotoxicity induced by 125 I-mIBG was quantitatively comparable by the peripheral rim thickness to that of 131 I-mIBG at the dose of 100 μCi. The peripheral rim thickness decreased most significantly in the first 17 hours after initial treatment. There was no statistical decrease in the rim thickness identified afterwards for the second, third, and fourth days of

  12. Homogeneous pancreatic cancer spheroids mimic growth pattern of circulating tumor cell clusters and macrometastases: displaying heterogeneity and crater-like structure on inner layer.

    Science.gov (United States)

    Feng, Hao; Ou, Bao-Chi; Zhao, Jing-Kun; Yin, Shuai; Lu, Ai-Guo; Oechsle, Eva; Thasler, Wolfgang E

    2017-09-01

    Pancreatic cancer 3D in vitro models including multicellular tumor spheroid (MCTS), single cell-derived tumor spheroid (SCTS), tissue-derived tumor spheroid, and organotypic models provided powerful platforms to mimic in vivo tumor. Recent work supports that circulating tumor cell (CTC) clusters are more efficient in metastasis seeding than single CTCs. The purpose of this study is to establish 3D culture models which can mimic single CTC, monoclonal CTC clusters, and the expansion of macrometastases. Seven pancreatic ductal adenocarcinoma cell lines were used to establish MCTS and SCTS using hanging drop and ultra-low attachment plates. Spheroid immunofluorescence staining, spheroid formation assay, immunoblotting, and literature review were performed to investigate molecular biomarkers and the morphological characteristics of pancreatic tumor spheroids. Single cells experienced different growth patterns to form SCTS, like signet ring-like cells, blastula-like structures, and solid core spheroids. However, golf ball-like hollow spheroids could also be detected, especially when DanG and Capan-1 cells were cultivated with fibroblast-conditioned medium (p cell lines could also establish tumor spheroid with hanging drop plates by adding methylated cellulose. Tumor spheroids derived from pancreatic cancer cell line DanG possessed asymmetrically distributed proliferation center, immune-checkpoint properties. ß-catenin, Ki-67, and F-actin were active surrounding the crater-like structure distributing on the inner layer of viable rim cover of the spheroids, which was relevant to well-differentiated tumor cells. It is possible to establish 3D CTC cluster models from homogenous PDA cell lines using hanging drop and ultra-low attachment plates. PDA cell line displays its own intrinsic properties or heterogeneity. The mechanism of formation of the crater-like structure as well as golf ball-like structure needs further exploration.

  13. Universal timescales in the rheology of spheroid cell aggregates

    Science.gov (United States)

    Yu, Miao; Mahtabfar, Aria; Beleen, Paul; Foty, Ramsey; Zahn, Jeffrey; Shreiber, David; Liu, Liping; Lin, Hao

    2017-11-01

    The rheological properties of tissue play important roles in key biological processes including embryogenesis, cancer metastasis, and wound healing. Spheroid cell aggregate is a particularly interesting model system for the study of these phenomena. In the long time, they behave like drops with a surface tension. In the short, viscoelasticity also needs to be considered. In this work, we discover two coupled and universal timescales for spheroid aggregates. A total of 12 aggregate types (total aggregate number n =290) derived from L and GBM (glioblastoma multiforme) cells are studied with microtensiometer to obtain their surface tension. They are also allowed to relax upon release of the compression forces. The two timescales are observed during the relaxation process; their values do not depend on compression time nor the degree of deformation, and are consistent among all 12 types. Following prior work (Yu et al., Phys. Rev. Lett., 115:128303; Liu et al., J. Mech. Phys. Solids, 98:309-329) we use a rigorous mathematical theory to interpret the results, which reveals intriguing properties of the aggregates on both tissue and cellular levels. The mechanics of multicellular organization reflects both complexity and regularity due to strong active regulation.

  14. RF induction plasma spheroidization of tungsten powders

    International Nuclear Information System (INIS)

    Gu Zhogntao; Ye Gaoying; Liu Chuandong; Tong Honghui

    2009-01-01

    Irregularly-shaped tungsten powders (average granular sizes of 512 μm) have been spheroidized by radio frequency (RF)induction plasma. The effects of feed rate, mode of material dispersion, particle size on spheroidization efficiency are investigated. Experimental results show that the spheroidization efficiency decreases rapidly when the feed rate increases to more than 95 g/min. Only 30% spheroidization efficiency is gained at the feed rate of 135.75 g/min. The spheroidization efficiency is also affected by the flow rate of carrier gas. When the flow rate of carrier gas is 0.12 m 3 /h, the dispersion effect is the best, and the spheroidization efficiency is almost 100%. The apparent density of tungsten powders increases a bit with the increase of spheroidization efficiency. And the particle size uniformity of spheroidized tungsten powders is in accordance with that of original powders. (authors)

  15. Bacterial biofilms: prokaryotic adventures in multicellularity

    DEFF Research Database (Denmark)

    Webb, J.S.; Givskov, Michael Christian; Kjelleberg, S.

    2003-01-01

    The development of bacterial biofilms includes both the initial social behavior of undifferentiated cells, as well as cell death and differentiation in the mature biofilm, and displays several striking similarities with higher organisms. Recent advances in the field provide new insight...... into differentiation and cell death events in bacterial biofilm development and propose that biofilms have an unexpected level of multicellularity....

  16. Role of multicellular aggregates in biofilm formation

    DEFF Research Database (Denmark)

    Kragh, Kasper N.; Hutchison, Jaime B.; Melaugh, Gavin

    2016-01-01

    In traditional models of in vitro biofilm development, individual bacterial cells seed a surface, multiply, and mature into multicellular, three-dimensional structures. Much research has been devoted to elucidating the mechanisms governing the initial attachment of single cells to surfaces. Howev...

  17. SIZE AND SHAPE FACTOR EXTREMES OF SPHEROIDS

    Directory of Open Access Journals (Sweden)

    Daniel Hlubinka

    2011-05-01

    Full Text Available In the paper we consider random prolate (oblate spheroids and their random profiles. The limiting distribution of the extremal characteristics of the spheroids is related to the limiting distribution of the corresponding extremal characteristics of the profiles. The difference between the analysis of the prolate and oblate spheroids is discussed. We propose the possible application of the theoretical results.

  18. A comparative study of post-irradiation growth kinetics of spheroids and monolayers

    International Nuclear Information System (INIS)

    Dertinger, J.; Luecke-Huhle, C.

    1975-01-01

    Post-irradiation growth kinetics of γ-irradiated spheroid and monolayer cells in exponential growth phase was investigated by means of dose-response curves based on cell counts after specified time intervals following irradiation. A mathematical model of cell-growth after irradiation was fitted to these curves. The model parameters (related to division delay and growth of non-surviving cells) obtained from this analysis consistently indicated increasing resistance to sub-lethal damage of cells cultured as multicellular spheroids under conditions of increasing three-dimensional contact. In contrast, no indication of an increased radiation-resistance was found with cells cultured on a substratum under a variety of conditions. (author)

  19. SMIFH2-mediated mDia formin functional inhibition potentiates chemotherapeutic targeting of human ovarian cancer spheroids.

    Science.gov (United States)

    Ziske, Megan A; Pettee, Krista M; Khaing, MaNada; Rubinic, Kaitlin; Eisenmann, Kathryn M

    2016-03-25

    Due to a lack of effective screening or prevention protocol for epithelial ovarian cancer (EOC), there is a critical unmet need to develop therapeutic interventions for EOC treatment. EOC metastasis is unique. Initial dissemination is not primarily hematogenous, yet is facilitated through shedding of primary tumor cells into the peritoneal fluid and accumulating ascites. Increasingly, isolated patient spheroids point to a clinical role for spheroids in EOC metastasis. EOC spheroids are highly invasive structures that disseminate upon peritoneal mesothelium, and visceral tissues including liver and omentum. Selection for this subset of chemoresistant EOC cells could influence disease progression and/or recurrence. Thus, targeting spheroid integrity/structure may improve the chemotherapeutic responsiveness of EOC. We discovered a critical role for mammalian Diaphanous (mDia)-related formin-2 in maintaining EOC spheroid structure. Both mDia2 and the related mDia1 regulate F-actin networks critical to maintain cell-cell contacts and the integrity of multi-cellular epithelial sheets. We investigated if mDia2 functional inhibition via a small molecule inhibitor SMIFH2 combined with chemotherapeutics, such as taxol and cisplatin, inhibits the viability of EOC monolayers and clinically relevant spheroids. SMIFH2-mediated mDia formin inhibition significantly reduced both ES2 and Skov3 EOC monolayer viability while spheroid viability was minimally impacted only at the highest concentrations. Combining either cisplatin or taxol with SMIFH2 did not significantly enhance the effects of either drug alone in ES2 monolayers, while Skov3 monolayers treated with taxol or cisplatin and SMIFH2 showed significant additive inhibition of viability. ES2 spheroids were highly responsive with clear additive anti-viability effects with dual taxol or cisplatin when combined with SMIFH2 treatments. While combined taxol with SMIFH2 in spheroids showed an additive effect relative to single

  20. Acoustic scattering on spheroidal shapes near boundaries

    Science.gov (United States)

    Miloh, Touvia

    2016-11-01

    A new expression for the Lamé product of prolate spheroidal wave functions is presented in terms of a distribution of multipoles along the axis of the spheroid between its foci (generalizing a corresponding theorem for spheroidal harmonics). Such an "ultimate" singularity system can be effectively used for solving various linear boundary-value problems governed by the Helmholtz equation involving prolate spheroidal bodies near planar or other boundaries. The general methodology is formally demonstrated for the axisymmetric acoustic scattering problem of a rigid (hard) spheroid placed near a hard/soft wall or inside a cylindrical duct under an axial incidence of a plane acoustic wave.

  1. Theory and computation of spheroidal wavefunctions

    International Nuclear Information System (INIS)

    Falloon, P E; Abbott, P C; Wang, J B

    2003-01-01

    In this paper we report on a package, written in the Mathematica computer algebra system, which has been developed to compute the spheroidal wavefunctions of Meixner and Schaefke (1954 Mathieusche Funktionen und Sphaeroidfunktionen) and is available online (physics.uwa.edu.au/~falloon/spheroidal/spheroidal.html). This package represents a substantial contribution to the existing software, since it computes the spheroidal wavefunctions to arbitrary precision for general complex parameters μ, ν, γ and argument z; existing software can only handle integer μ, ν and does not give arbitrary precision. The package also incorporates various special cases and computes analytic power series and asymptotic expansions in the parameter γ. The spheroidal wavefunctions of Flammer (1957 Spheroidal Wave functions) are included as a special case of Meixner's more general functions. This paper presents a concise review of the general theory of spheroidal wavefunctions and a description of the formulae and algorithms used in their computation, and gives high precision numerical examples

  2. A Model for Spheroid versus Monolayer Response of SK-N-SH Neuroblastoma Cells to Treatment with 15-Deoxy-PGJ2

    Directory of Open Access Journals (Sweden)

    Dorothy I. Wallace

    2016-01-01

    Full Text Available Researchers have observed that response of tumor cells to treatment varies depending on whether the cells are grown in monolayer, as in vitro spheroids or in vivo. This study uses data from the literature on monolayer treatment of SK-N-SH neuroblastoma cells with 15-deoxy-PGJ2 and couples it with data on growth rates for untreated SK-N-SH neuroblastoma cells grown as multicellular spheroids. A linear model is constructed for untreated and treated monolayer data sets, which is tuned to growth, death, and cell cycle data for the monolayer case for both control and treatment with 15-deoxy-PGJ2. The monolayer model is extended to a five-dimensional nonlinear model of in vitro tumor spheroid growth and treatment that includes compartments of the cell cycle (G1,S,G2/M as well as quiescent (Q and necrotic (N cells. Monolayer treatment data for 15-deoxy-PGJ2 is used to derive a prediction of spheroid response under similar treatments. For short periods of treatment, spheroid response is less pronounced than monolayer response. The simulations suggest that the difference in response to treatment of monolayer versus spheroid cultures observed in laboratory studies is a natural consequence of tumor spheroid physiology rather than any special resistance to treatment.

  3. Extracellular signaling and multicellularity in Bacillus subtilis.

    Science.gov (United States)

    Shank, Elizabeth Anne; Kolter, Roberto

    2011-12-01

    Bacillus subtilis regulates its ability to differentiate into distinct, co-existing cell types in response to extracellular signaling molecules produced either by itself, or present in its environment. The production of molecules by B. subtilis cells, as well as their response to these signals, is not uniform across the population. There is specificity and heterogeneity both within genetically identical populations as well as at the strain-level and species-level. This review will discuss how extracellular signaling compounds influence B. subtilis multicellularity with regard to matrix-producing cannibal differentiation, germination, and swarming behavior, as well as the specificity of the quorum-sensing peptides ComX and CSF. It will also highlight how imaging mass spectrometry can aid in identifying signaling compounds and contribute to our understanding of the functional relationship between such compounds and multicellular behavior. Copyright © 2011 Elsevier Ltd. All rights reserved.

  4. PhysiCell: An open source physics-based cell simulator for 3-D multicellular systems

    Science.gov (United States)

    Ghaffarizadeh, Ahmadreza; Mumenthaler, Shannon M.

    2018-01-01

    Many multicellular systems problems can only be understood by studying how cells move, grow, divide, interact, and die. Tissue-scale dynamics emerge from systems of many interacting cells as they respond to and influence their microenvironment. The ideal “virtual laboratory” for such multicellular systems simulates both the biochemical microenvironment (the “stage”) and many mechanically and biochemically interacting cells (the “players” upon the stage). PhysiCell—physics-based multicellular simulator—is an open source agent-based simulator that provides both the stage and the players for studying many interacting cells in dynamic tissue microenvironments. It builds upon a multi-substrate biotransport solver to link cell phenotype to multiple diffusing substrates and signaling factors. It includes biologically-driven sub-models for cell cycling, apoptosis, necrosis, solid and fluid volume changes, mechanics, and motility “out of the box.” The C++ code has minimal dependencies, making it simple to maintain and deploy across platforms. PhysiCell has been parallelized with OpenMP, and its performance scales linearly with the number of cells. Simulations up to 105-106 cells are feasible on quad-core desktop workstations; larger simulations are attainable on single HPC compute nodes. We demonstrate PhysiCell by simulating the impact of necrotic core biomechanics, 3-D geometry, and stochasticity on the dynamics of hanging drop tumor spheroids and ductal carcinoma in situ (DCIS) of the breast. We demonstrate stochastic motility, chemical and contact-based interaction of multiple cell types, and the extensibility of PhysiCell with examples in synthetic multicellular systems (a “cellular cargo delivery” system, with application to anti-cancer treatments), cancer heterogeneity, and cancer immunology. PhysiCell is a powerful multicellular systems simulator that will be continually improved with new capabilities and performance improvements. It also

  5. PhysiCell: An open source physics-based cell simulator for 3-D multicellular systems.

    Science.gov (United States)

    Ghaffarizadeh, Ahmadreza; Heiland, Randy; Friedman, Samuel H; Mumenthaler, Shannon M; Macklin, Paul

    2018-02-01

    Many multicellular systems problems can only be understood by studying how cells move, grow, divide, interact, and die. Tissue-scale dynamics emerge from systems of many interacting cells as they respond to and influence their microenvironment. The ideal "virtual laboratory" for such multicellular systems simulates both the biochemical microenvironment (the "stage") and many mechanically and biochemically interacting cells (the "players" upon the stage). PhysiCell-physics-based multicellular simulator-is an open source agent-based simulator that provides both the stage and the players for studying many interacting cells in dynamic tissue microenvironments. It builds upon a multi-substrate biotransport solver to link cell phenotype to multiple diffusing substrates and signaling factors. It includes biologically-driven sub-models for cell cycling, apoptosis, necrosis, solid and fluid volume changes, mechanics, and motility "out of the box." The C++ code has minimal dependencies, making it simple to maintain and deploy across platforms. PhysiCell has been parallelized with OpenMP, and its performance scales linearly with the number of cells. Simulations up to 105-106 cells are feasible on quad-core desktop workstations; larger simulations are attainable on single HPC compute nodes. We demonstrate PhysiCell by simulating the impact of necrotic core biomechanics, 3-D geometry, and stochasticity on the dynamics of hanging drop tumor spheroids and ductal carcinoma in situ (DCIS) of the breast. We demonstrate stochastic motility, chemical and contact-based interaction of multiple cell types, and the extensibility of PhysiCell with examples in synthetic multicellular systems (a "cellular cargo delivery" system, with application to anti-cancer treatments), cancer heterogeneity, and cancer immunology. PhysiCell is a powerful multicellular systems simulator that will be continually improved with new capabilities and performance improvements. It also represents a significant

  6. Chondroitin sulfate microparticles modulate transforming growth factor-β1-induced chondrogenesis of human mesenchymal stem cell spheroids.

    Science.gov (United States)

    Goude, Melissa C; McDevitt, Todd C; Temenoff, Johnna S

    2014-01-01

    Mesenchymal stem cells (MSCs) have been previously explored as a part of cell-based therapies for the repair of damaged cartilage. Current MSC chondrogenic differentiation strategies employ large pellets; however, we have developed a technique to form small MSC aggregates (500-1,000 cells) that can reduce transport barriers while maintaining a multicellular structure analogous to cartilaginous condensations. The objective of this study was to examine the effects of incorporating chondroitin sulfate methacrylate (CSMA) microparticles (MPs) within small MSC spheroids cultured in the presence of transforming growth factor (TGF)-β1 on chondrogenesis. Spheroids with MPs induced earlier increases in collagen II and aggrecan gene expression (chondrogenic markers) than spheroids without MPs, although no large differences in immunostaining for these matrix molecules were observed by day 21 between these groups. Collagen I and X were also detected in the extracellular matrix (ECM) of all spheroids by immunostaining. Interestingly, histology revealed that CSMA MPs clustered together near the center of the MSC spheroids and induced circumferential alignment of cells and ECM around the material core. This study demonstrates the use of CSMA materials to further examine the effects of matrix molecules on MSC phenotype as well as potentially direct differentiation in a more spatially controlled manner that better mimics the architecture of specific musculoskeletal tissues. © 2014 S. Karger AG, Basel.

  7. Effect of apoptosis and response of extracellular matrix proteins after chemotherapy application on human breast cancer cell spheroids.

    Science.gov (United States)

    Oktem, G; Vatansever, S; Ayla, S; Uysal, A; Aktas, S; Karabulut, B; Bilir, A

    2006-02-01

    Multicellular tumor spheroid (MTS) represents a three-dimensional structural form of tumors in laboratory conditions, and it has the characteristics of avascular micrometastases or intervascular spaces of big tumors. Recent studies indicate that extracellular matrix (ECM) proteins play a critical role in tumor metastasis, therefore normal and cancer cells require an ECM for survival, proliferation and differentiation. Doxorubicin and Docetaxel are widely used in the therapy of breast cancer, as well as in in vivo and in vitro studies. In this study, we examined the effect of apoptosis and proliferation of cells on the human breast cancer cell line, MCF-7, by using p53, bcl-2 and Ki67 gene expression, and the tendency to metastasis with extracellular matrix proteins, laminin and type IV collagen after chemotherapy in the spheroid model. The apoptotic cell death in situ was detected by TUNEL method. TUNEL-positive cells and positive immunoreactivities of laminin, type IV collagen, p53 and, bcl-2 were detected in the control group. There was no laminin and type IV collagen immunoreactivities in spheroids of drug groups. While TUNEL-positive cells and p53 immunoreactivity were detected in Docetaxel, Doxorubicin and Docetaxel/Doxorubicin groups, p53 immunoreactivity was not observed in the Docetaxel group. There was no bcl-2 immunoreactivity in either drug group. In addition, we did not detect Ki67 immunoreactivity in both control and drug treatment groups. However, the absence of Ki67 protein in MCF-7 breast multicellular tumor spheroids is possibly related to the cells in G0 or S phase. These chemotherapeutic agents may affect the presence of ECM proteins in this in vitro model of micrometastasis of spheroids. These findings suggest that the possible mechanism of cell death in Doxorubicin and Docetaxel/Doxorubicin treatment groups is related to apoptosis through the p53 pathway. However, we considered the possibility that there is another control mechanism for the

  8. Spheroid Culture of Head and Neck Cancer Cells Reveals an Important Role of EGFR Signalling in Anchorage Independent Survival.

    Science.gov (United States)

    Braunholz, Diana; Saki, Mohammad; Niehr, Franziska; Öztürk, Merve; Borràs Puértolas, Berta; Konschak, Robert; Budach, Volker; Tinhofer, Ingeborg

    2016-01-01

    In solid tumours millions of cells are shed into the blood circulation each day. Only a subset of these circulating tumour cells (CTCs) survive, many of them presumable because of their potential to form multi-cellular clusters also named spheroids. Tumour cells within these spheroids are protected from anoikis, which allows them to metastasize to distant organs or re-seed at the primary site. We used spheroid cultures of head and neck squamous cell carcinoma (HNSCC) cell lines as a model for such CTC clusters for determining the role of the epidermal growth factor receptor (EGFR) in cluster formation ability and cell survival after detachment from the extra-cellular matrix. The HNSCC cell lines FaDu, SCC-9 and UT-SCC-9 (UT-SCC-9P) as well as its cetuximab (CTX)-resistant sub-clone (UT-SCC-9R) were forced to grow in an anchorage-independent manner by coating culture dishes with the anti-adhesive polymer poly-2-hydroxyethylmethacrylate (poly-HEMA). The extent of apoptosis, clonogenic survival and EGFR signalling under such culture conditions was evaluated. The potential of spheroid formation in suspension culture was found to be positively correlated with the proliferation rate of HNSCC cell lines as well as their basal EGFR expression levels. CTX and gefitinib blocked, whereas the addition of EGFR ligands promoted anchorage-independent cell survival and spheroid formation. Increased spheroid formation and growth were associated with persistent activation of EGFR and its downstream signalling component (MAPK/ERK). Importantly, HNSCC cells derived from spheroid cultures retained their clonogenic potential in the absence of cell-matrix contact. Addition of CTX under these conditions strongly inhibited colony formation in CTX-sensitive cell lines but not their resistant subclones. Altogether, EGFR activation was identified as crucial factor for anchorage-independent survival of HNSCC cells. Targeting EGFR in CTC cluster formation might represent an attractive anti

  9. Scalable robotic biofabrication of tissue spheroids

    International Nuclear Information System (INIS)

    Mehesz, A Nagy; Hajdu, Z; Visconti, R P; Markwald, R R; Mironov, V; Brown, J; Beaver, W; Da Silva, J V L

    2011-01-01

    Development of methods for scalable biofabrication of uniformly sized tissue spheroids is essential for tissue spheroid-based bioprinting of large size tissue and organ constructs. The most recent scalable technique for tissue spheroid fabrication employs a micromolded recessed template prepared in a non-adhesive hydrogel, wherein the cells loaded into the template self-assemble into tissue spheroids due to gravitational force. In this study, we present an improved version of this technique. A new mold was designed to enable generation of 61 microrecessions in each well of a 96-well plate. The microrecessions were seeded with cells using an EpMotion 5070 automated pipetting machine. After 48 h of incubation, tissue spheroids formed at the bottom of each microrecession. To assess the quality of constructs generated using this technology, 600 tissue spheroids made by this method were compared with 600 spheroids generated by the conventional hanging drop method. These analyses showed that tissue spheroids fabricated by the micromolded method are more uniform in diameter. Thus, use of micromolded recessions in a non-adhesive hydrogel, combined with automated cell seeding, is a reliable method for scalable robotic fabrication of uniform-sized tissue spheroids.

  10. Scalable robotic biofabrication of tissue spheroids

    Energy Technology Data Exchange (ETDEWEB)

    Mehesz, A Nagy; Hajdu, Z; Visconti, R P; Markwald, R R; Mironov, V [Advanced Tissue Biofabrication Center, Department of Regenerative Medicine and Cell Biology, Medical University of South Carolina, Charleston, SC (United States); Brown, J [Department of Mechanical Engineering, Clemson University, Clemson, SC (United States); Beaver, W [York Technical College, Rock Hill, SC (United States); Da Silva, J V L, E-mail: mironovv@musc.edu [Renato Archer Information Technology Center-CTI, Campinas (Brazil)

    2011-06-15

    Development of methods for scalable biofabrication of uniformly sized tissue spheroids is essential for tissue spheroid-based bioprinting of large size tissue and organ constructs. The most recent scalable technique for tissue spheroid fabrication employs a micromolded recessed template prepared in a non-adhesive hydrogel, wherein the cells loaded into the template self-assemble into tissue spheroids due to gravitational force. In this study, we present an improved version of this technique. A new mold was designed to enable generation of 61 microrecessions in each well of a 96-well plate. The microrecessions were seeded with cells using an EpMotion 5070 automated pipetting machine. After 48 h of incubation, tissue spheroids formed at the bottom of each microrecession. To assess the quality of constructs generated using this technology, 600 tissue spheroids made by this method were compared with 600 spheroids generated by the conventional hanging drop method. These analyses showed that tissue spheroids fabricated by the micromolded method are more uniform in diameter. Thus, use of micromolded recessions in a non-adhesive hydrogel, combined with automated cell seeding, is a reliable method for scalable robotic fabrication of uniform-sized tissue spheroids.

  11. Inhibition of hexokinase-2 with targeted liposomal 3-bromopyruvate in an ovarian tumor spheroid model of aerobic glycolysis

    Directory of Open Access Journals (Sweden)

    Gandham SK

    2015-07-01

    Full Text Available Srujan Kumar Gandham, Meghna Talekar, Amit Singh, Mansoor M Amiji Department of Pharmaceutical Sciences, School of Pharmacy, Northeastern University, Boston, MA, USA Background: The objective of this study was to evaluate the expression levels of glycolytic markers, especially hexokinase-2 (HK2, using a three-dimensional multicellular spheroid model of human ovarian adenocarcinoma (SKOV-3 cells and to develop an epidermal growth factor receptor-targeted liposomal formulation for improving inhibition of HK2 and the cytotoxicity of 3-bromopyruvate (3-BPA. Methods: Multicellular SKOV-3 tumor spheroids were developed using the hanging drop method and expression levels of glycolytic markers were examined. Non-targeted and epidermal growth factor receptor-targeted liposomal formulations of 3-BPA were formulated and characterized. Permeability and cellular uptake of the liposomal formulations in three-dimensional SKOV-3 spheroids was evaluated using confocal microscopy. The cytotoxicity and HK2 inhibition potential of solution form of 3-BPA was compared to the corresponding liposomal formulation by using cell proliferation and HK2 enzymatic assays. Results: SKOV-3 spheroids were reproducibly developed using the 96-well hanging drop method, with an average size of 900 µm by day 5. HK2 enzyme activity levels under hypoxic conditions were found to be higher than under normoxic conditions (P<0.0001, Student’s t-test, unpaired and two-tailed. Liposomal formulations (both non-targeted and targeted of 3-BPA showed a more potent inhibitory effect (P<0.001, Student’s t-test, unpaired and two-tailed at a dose of 50 µM than the aqueous solution form at 3, 6, and 24 hours post administration. Similarly, the cytotoxic activity 3-BPA at various concentrations (10 µM–100 µM showed that the liposomal formulations had an enhanced cytotoxic effect of 2–5-fold (P<0.0001, Student’s t-test, unpaired and two-tailed when compared to the aqueous solution form

  12. Inhibition of hexokinase-2 with targeted liposomal 3-bromopyruvate in an ovarian tumor spheroid model of aerobic glycolysis.

    Science.gov (United States)

    Gandham, Srujan Kumar; Talekar, Meghna; Singh, Amit; Amiji, Mansoor M

    2015-01-01

    The objective of this study was to evaluate the expression levels of glycolytic markers, especially hexokinase-2 (HK2), using a three-dimensional multicellular spheroid model of human ovarian adenocarcinoma (SKOV-3) cells and to develop an epidermal growth factor receptor-targeted liposomal formulation for improving inhibition of HK2 and the cytotoxicity of 3-bromopyruvate (3-BPA). Multicellular SKOV-3 tumor spheroids were developed using the hanging drop method and expression levels of glycolytic markers were examined. Non-targeted and epidermal growth factor receptor-targeted liposomal formulations of 3-BPA were formulated and characterized. Permeability and cellular uptake of the liposomal formulations in three-dimensional SKOV-3 spheroids was evaluated using confocal microscopy. The cytotoxicity and HK2 inhibition potential of solution form of 3-BPA was compared to the corresponding liposomal formulation by using cell proliferation and HK2 enzymatic assays. SKOV-3 spheroids were reproducibly developed using the 96-well hanging drop method, with an average size of 900 µm by day 5. HK2 enzyme activity levels under hypoxic conditions were found to be higher than under normoxic conditions (P<0.0001, Student's t-test, unpaired and two-tailed). Liposomal formulations (both non-targeted and targeted) of 3-BPA showed a more potent inhibitory effect (P<0.001, Student's t-test, unpaired and two-tailed) at a dose of 50 µM than the aqueous solution form at 3, 6, and 24 hours post administration. Similarly, the cytotoxic activity 3-BPA at various concentrations (10 µM-100 µM) showed that the liposomal formulations had an enhanced cytotoxic effect of 2-5-fold (P<0.0001, Student's t-test, unpaired and two-tailed) when compared to the aqueous solution form for both 10 µM and 25 µM concentrations. SKOV-3 spheroids developed by the hanging drop method can be used as a tumor aerobic glycolysis model for evaluation of therapies targeting the glycolytic pathway in cancer

  13. The distribution of alternative agents for targeted radiotherapy within human neuroblastoma spheroids

    International Nuclear Information System (INIS)

    Mairs, R.J.; Gaze, M.N.; Murray, T.; Reid, R.; McSharry, C.; Babich, J.W.

    1991-01-01

    This study aims to select the radiopharmaceutical vehicle for targeted radiotherapy of neuroblastoma which is most likely to penetrate readily the centre of micrometastases in vivo. The human neuroblastoma cell line NB1-G, grown as multicellular spheroids provided an in vitro model for micrometastases. The radiopharmaceuticals studied were the catecholamine analogue metaiodobenzyl guanidine (mIBG), a specific neuroectodermal monoclonal antibody (UJ13A) and β nerve growth factor (βNGF). Following incubation of each drug with neuroblastoma spheroids, autoradiographs of frozen sections were prepared to demonstrate their relative distributions. mIBG and βNGF were found to penetrate the centre of spheroids readily although the concentration of mIBG greatly exceeded that of βNGF. In contrast, UJ13A was only bound peripherally. We conclude that mIBG is the best available vehicle for targeted radiotherapy of neuroblastoma cells with active uptake mechanisms for catecholimines. It is suggested that radionuclides with a shorter range of emissions than 131 I may be conjugated to benzyl guanidine to constitute more effective targeting agents with potentially less toxicity to adjacent normal tissues. (author)

  14. Role of Multicellular Aggregates in Biofilm Formation

    Directory of Open Access Journals (Sweden)

    Kasper N. Kragh

    2016-03-01

    Full Text Available In traditional models of in vitro biofilm development, individual bacterial cells seed a surface, multiply, and mature into multicellular, three-dimensional structures. Much research has been devoted to elucidating the mechanisms governing the initial attachment of single cells to surfaces. However, in natural environments and during infection, bacterial cells tend to clump as multicellular aggregates, and biofilms can also slough off aggregates as a part of the dispersal process. This makes it likely that biofilms are often seeded by aggregates and single cells, yet how these aggregates impact biofilm initiation and development is not known. Here we use a combination of experimental and computational approaches to determine the relative fitness of single cells and preformed aggregates during early development of Pseudomonas aeruginosa biofilms. We find that the relative fitness of aggregates depends markedly on the density of surrounding single cells, i.e., the level of competition for growth resources. When competition between aggregates and single cells is low, an aggregate has a growth disadvantage because the aggregate interior has poor access to growth resources. However, if competition is high, aggregates exhibit higher fitness, because extending vertically above the surface gives cells at the top of aggregates better access to growth resources. Other advantages of seeding by aggregates, such as earlier switching to a biofilm-like phenotype and enhanced resilience toward antibiotics and immune response, may add to this ecological benefit. Our findings suggest that current models of biofilm formation should be reconsidered to incorporate the role of aggregates in biofilm initiation.

  15. Analysis of motility in multicellular Chlamydomonas reinhardtii evolved under predation.

    Directory of Open Access Journals (Sweden)

    Margrethe Boyd

    Full Text Available The advent of multicellularity was a watershed event in the history of life, yet the transition from unicellularity to multicellularity is not well understood. Multicellularity opens up opportunities for innovations in intercellular communication, cooperation, and specialization, which can provide selective advantages under certain ecological conditions. The unicellular alga Chlamydomonas reinhardtii has never had a multicellular ancestor yet it is closely related to the volvocine algae, a clade containing taxa that range from simple unicells to large, specialized multicellular colonies. Simple multicellular structures have been observed to evolve in C. reinhardtii in response to predation or to settling rate-based selection. Structures formed in response to predation consist of individual cells confined within a shared transparent extracellular matrix. Evolved isolates form such structures obligately under culture conditions in which their wild type ancestors do not, indicating that newly-evolved multicellularity is heritable. C. reinhardtii is capable of photosynthesis, and possesses an eyespot and two flagella with which it moves towards or away from light in order to optimize input of radiant energy. Motility contributes to C. reinhardtii fitness because it allows cells or colonies to achieve this optimum. Utilizing phototaxis to assay motility, we determined that newly evolved multicellular strains do not exhibit significant directional movement, even though the flagellae of their constituent unicells are present and active. In C. reinhardtii the first steps towards multicellularity in response to predation appear to result in a trade-off between motility and differential survivorship, a trade-off that must be overcome by further genetic change to ensure long-term success of the new multicellular organism.

  16. Intrinsically Disordered Proteins and the Origins of Multicellular Organisms

    Science.gov (United States)

    Dunker, A. Keith

    In simple multicellular organisms all of the cells are in direct contact with the surrounding milieu, whereas in complex multicellular organisms some cells are completely surrounded by other cells. Current phylogenetic trees indicate that complex multicellular organisms evolved independently from unicellular ancestors about 10 times, and only among the eukaryotes, including once for animals, twice each for green, red, and brown algae, and thrice for fungi. Given these multiple independent evolutionary lineages, we asked two questions: 1. Which molecular functions underpinned the evolution of multicellular organisms?; and, 2. Which of these molecular functions depend on intrinsically disordered proteins (IDPs)? Compared to unicellularity, multicellularity requires the advent of molecules for cellular adhesion, for cell-cell communication and for developmental programs. In addition, the developmental programs need to be regulated over space and time. Finally, each multicellular organism has cell-specific biochemistry and physiology. Thus, the evolution of complex multicellular organisms from unicellular ancestors required five new classes of functions. To answer the second question we used Key-words in Swiss Protein ranked for associations with predictions of protein structure or disorder. With a Z-score of 18.8 compared to random-function proteins, à differentiation was the biological process most strongly associated with IDPs. As expected from this result, large numbers of individual proteins associated with differentiation exhibit substantial regions of predicted disorder. For the animals for which there is the most readily available data all five of the underpinning molecular functions for multicellularity were found to depend critically on IDP-based mechanisms and other evidence supports these ideas. While the data are more sparse, IDPs seem to similarly underlie the five new classes of functions for plants and fungi as well, suggesting that IDPs were indeed

  17. Translation and rotation of a porous spheroid in a spheroidal container

    International Nuclear Information System (INIS)

    Saad, E.I.

    2010-01-01

    The flow problem of an incompressible axisymmetrical quasisteady translation and steady rotation of a porous spheroid in a concentric spheroidal container are studied analytically. The same small departure from a sphere is considered for each spheroidal surface. In the limit of small Reynolds number, the Brinkman equation for the flow inside the porous region and the Stokes equation for the outside region in their stream functions formulations and velocity components, which are proportional to the translational and angular velocities, respectively, are used. Explicit expressions are obtained for both inside and outside flow fields to the first order in a small parameter characterizing the deformation of the spheroidal surface from the spherical shape. The hydrodynamic drag force and couple exerted on the porous spheroid are obtained for the special cases of prolate and oblate spheroids in closed forms. The dependence of the normalized wall-corrected translational and rotational mobilities on permeability for a porous spheroid in an unbounded medium and for a solid spheroid in a cell on the particle volume fraction is discussed numerically and graphically for various values of the deformation parameter. In the limiting cases, the analytical solutions describing the drag force and torque or mobilities for a porous spheroid in the spheroidal vessel reduce to those for a solid sphere and for a porous sphere in a spherical cell. (author)

  18. Solution of diffusion equation in deformable spheroids

    Energy Technology Data Exchange (ETDEWEB)

    Ayyoubzadeh, Seyed Mohsen [Department of Energy Engineering, Sharif University of Technology, Tehran (Iran, Islamic Republic of); Safari, Mohammad Javad, E-mail: iFluka@gmail.com [Department of Energy Engineering, Sharif University of Technology, Tehran (Iran, Islamic Republic of); Vosoughi, Naser [Department of Energy Engineering, Sharif University of Technology, Tehran (Iran, Islamic Republic of)

    2011-05-15

    Research highlights: > Developing an explicit solution for the diffusion equation in spheroidal geometry. > Proving an orthogonality relation for spheroidal eigenfunctions. > Developing a relation for the extrapolation distance in spheroidal geometry. > Considering the sphere and slab as limiting cases for a spheroid. > Cross-validation of the analytical solution with Monte Carlo simulations. - Abstract: The time-dependent diffusion of neutrons in a spheroid as a function of the focal distance has been studied. The solution is based on an orthogonal basis and an extrapolation distanced related boundary condition for the spheroidal geometry. It has been shown that spheres and disks are two limiting cases for the spheroids, for which there is a smooth transition for the systems properties between these two limits. Furthermore, it is demonstrated that a slight deformation from a sphere does not affect the fundamental mode properties, to the first order. The calculations for both multiplying and non-multiplying media have been undertaken, showing good agreement with direct Monte Carlo simulations.

  19. Charged fluid distribution in higher dimensional spheroidal space-time

    Indian Academy of Sciences (India)

    associated 3-spaces obtained as hypersurfaces t = constant, 3-spheroids, are suit- ... pressure. Considering the Vaidya–Tikekar [12] spheroidal geometry, ... a relativistic star in hydrostatic equilibrium having the spheroidal geometry of the .... K = 1, the spheroidal 3-space degenerates into a flat 3-space and when K = 0 it.

  20. Global gene expression analysis of early response to chemotherapy treatment in ovarian cancer spheroids

    Directory of Open Access Journals (Sweden)

    Tetu Bernard

    2008-02-01

    Full Text Available Abstract Background Chemotherapy (CT resistance in ovarian cancer (OC is broad and encompasses diverse unrelated drugs, suggesting more than one mechanism of resistance. To better understand the molecular mechanisms controlling the immediate response of OC cells to CT exposure, we have performed gene expression profiling in spheroid cultures derived from six OC cell lines (OVCAR3, SKOV3, TOV-112, TOV-21, OV-90 and TOV-155, following treatment with 10,0 μM cisplatin, 2,5 μM paclitaxel or 5,0 μM topotecan for 72 hours. Results Exposure of OC spheroids to these CT drugs resulted in differential expression of genes associated with cell growth and proliferation, cellular assembly and organization, cell death, cell cycle control and cell signaling. Genes, functionally involved in DNA repair, DNA replication and cell cycle arrest were mostly overexpressed, while genes implicated in metabolism (especially lipid metabolism, signal transduction, immune and inflammatory response, transport, transcription regulation and protein biosynthesis, were commonly suppressed following all treatments. Cisplatin and topotecan treatments triggered similar alterations in gene and pathway expression patterns, while paclitaxel action was mainly associated with induction of genes and pathways linked to cellular assembly and organization (including numerous tubulin genes, cell death and protein synthesis. The microarray data were further confirmed by pathway and network analyses. Conclusion Most alterations in gene expression were directly related to mechanisms of the cytotoxics actions in OC spheroids. However, the induction of genes linked to mechanisms of DNA replication and repair in cisplatin- and topotecan-treated OC spheroids could be associated with immediate adaptive response to treatment. Similarly, overexpression of different tubulin genes upon exposure to paclitaxel could represent an early compensatory effect to this drug action. Finally, multicellular

  1. Bar-spheroid interaction in galaxies

    Science.gov (United States)

    Hernquist, Lars; Weinberg, Martin D.

    1992-01-01

    N-body simulation and linear analysis is employed to investigate the secular evolution of barred galaxies, with emphasis on the interaction between bars and spheroidal components of galaxies. This interaction is argued to drive secular transfer of angular momentum from bars to spheroids, primarily through resonant coupling. A moderately strong bar, having mass within corotation about 0.3 times the enclosed spheroid mass, is predicted to shed all its angular momentum typically in less than about 10 exp 9 yr. Even shorter depletion time scales are found for relatively more massive bars. It is suggested either that spheroids around barred galaxies are structured so as to inhibit strong coupling with bars, or that bars can form by unknown processes long after disks are established. The present models reinforce the notion that bars can drive secular evolution in galaxies.

  2. Plasma spheroidization and cladding of powders

    Energy Technology Data Exchange (ETDEWEB)

    Petrunichev, V.A.; Averin, V.V.; Sorokin, L.M.; Koroleva, E.B.

    1987-02-01

    With reference to experimental results for nickel and chromium alloys, it is shown that complex alloy powders can be spheroidized in plasma discharges using an argon plasma with hydrogen. The spheroidizing process is accompanied by the reduction of surface oxides, with uniform element distribution within the particles; the granulometric composition of the particles is preserved. It is also shown that plasma technology can be used for producing metal-clad oxide and carbide powders, which improve the performance of cermets and coatings.

  3. How the evolution of multicellularity set the stage for cancer

    Science.gov (United States)

    Trigos, Anna S; Pearson, Richard B; Papenfuss, Anthony T; Goode, David L

    2018-01-01

    Neoplastic growth and many of the hallmark properties of cancer are driven by the disruption of molecular networks established during the emergence of multicellularity. Regulatory pathways and molecules that evolved to impose regulatory constraints upon networks established in earlier unicellular organisms enabled greater communication and coordination between the diverse cell types required for multicellularity, but also created liabilities in the form of points of vulnerability in the network that when mutated or dysregulated facilitate the development of cancer. These factors are usually overlooked in genomic analyses of cancer, but understanding where vulnerabilities to cancer lie in the networks of multicellular species would provide important new insights into how core molecular processes and gene regulation change during tumourigenesis. We describe how the evolutionary origins of genes influence their roles in cancer, and how connections formed between unicellular and multicellular genes that act as key regulatory hubs for normal tissue homeostasis can also contribute to malignant transformation when disrupted. Tumours in general are characterised by increased dependence on unicellular processes for survival, and major dysregulation of the control structures imposed on these processes during the evolution of multicellularity. Mounting molecular evidence suggests altered interactions at the interface between unicellular and multicellular genes play key roles in the initiation and progression of cancer. Furthermore, unicellular network regions activated in cancer show high degrees of robustness and plasticity, conferring increased adaptability to tumour cells by supporting effective responses to environmental pressures such as drug exposure. Examining how the links between multicellular and unicellular regions get disrupted in tumours has great potential to identify novel drivers of cancer, and to guide improvements to cancer treatment by identifying more

  4. Repair during multifraction exposures: spheroids versus monolayers

    International Nuclear Information System (INIS)

    Durand, R.E.

    1984-01-01

    Many type of mammalian cells, when grown in culture as multicell spheroids, display an increased ability to accumulate and repair sublethal radiation damage which has been called the ''contact effect''. Since this effect has the potential to markedly modify the multifraction radiation response of cells in V79 spheroids relative to cells in monolayer cultures, an investigation was made of regimens ranging from 1 to 100 fractions. Effective dose rates were chosen near 1 Gy h -1 to inhibit cell progression and thus simplify analysis of the results. As expected, larger doses per fraction produced more net cell killing in both systems than lower doses per fraction. Additionally, less killing of spheroid cells was observed in all regimens, in accord with their greater potential for repair. However, when the data were expressed as isoeffect curves, the spheroid and monolayer curves converged as the number of fractions increased. Thus, quite similar inherent sensitivity and repair capabilities would be predicted for ultra-low doses per fraction. High precision techniques for defining survival after doses of radiation from 0.2 to 1 Gy were, however, still able to demonstrate a survival advantage for cells grown as spheroids. (author)

  5. Bistable flows in precessing spheroids

    Energy Technology Data Exchange (ETDEWEB)

    Cébron, D, E-mail: david.cebron@ujf-grenoble.fr [Université Grenoble Alpes, CNRS, ISTerre, Grenoble (France)

    2015-04-15

    Precession driven flows are found in any rotating container filled with liquid, when the rotation axis itself rotates about a secondary axis that is fixed in an inertial frame of reference. Because of its relevance for planetary fluid layers, many works consider spheroidal containers, where the uniform vorticity component of the bulk flow is reliably given by the well-known equations obtained by Busse (1968 J. Fluid Mech. 33 739–51). So far however, no analytical result for the solutions is available. Moreover, the cases where multiple flows can coexist have not been investigated in detail since their discovery by Noir et al (2003 Geophys. J. Int. 154 407–16). In this work we aim at deriving analytical results for the solutions, aiming in particular at first estimating the ranges of parameters where multiple solutions exist, and second studying quantitatively their stability. Using the models recently proposed by Noir and Cébron (2013 J. Fluid Mech. 737 412–39), which are more generic in the inviscid limit than the equations of Busse, we analytically describe these solutions, their conditions of existence, and their stability in a systematic manner. We then successfully compare these analytical results with the theory of Busse (1968). Dynamical model equations are finally proposed to investigate the stability of the solutions, which describe the bifurcation of the unstable flow solution. We also report for the first time the possibility that time-dependent multiple flows can coexist in precessing triaxial ellipsoids. Numerical integrations of the algebraic and differential equations have been efficiently performed with the dedicated script FLIPPER (supplementary material). (paper)

  6. Towards personalized computational oncology: from spatial models of tumour spheroids, to organoids, to tissues.

    Science.gov (United States)

    Karolak, Aleksandra; Markov, Dmitry A; McCawley, Lisa J; Rejniak, Katarzyna A

    2018-01-01

    A main goal of mathematical and computational oncology is to develop quantitative tools to determine the most effective therapies for each individual patient. This involves predicting the right drug to be administered at the right time and at the right dose. Such an approach is known as precision medicine. Mathematical modelling can play an invaluable role in the development of such therapeutic strategies, since it allows for relatively fast, efficient and inexpensive simulations of a large number of treatment schedules in order to find the most effective. This review is a survey of mathematical models that explicitly take into account the spatial architecture of three-dimensional tumours and address tumour development, progression and response to treatments. In particular, we discuss models of epithelial acini, multicellular spheroids, normal and tumour spheroids and organoids, and multi-component tissues. Our intent is to showcase how these in silico models can be applied to patient-specific data to assess which therapeutic strategies will be the most efficient. We also present the concept of virtual clinical trials that integrate standard-of-care patient data, medical imaging, organ-on-chip experiments and computational models to determine personalized medical treatment strategies. © 2018 The Author(s).

  7. Misorientations in spheroidal graphite: some new insights about spheroidal graphite growth in cast irons

    International Nuclear Information System (INIS)

    Lacaze, J; Theuwissen, K; Laffont, L; Véron, M

    2016-01-01

    Local diffraction patterning, orientation mapping and high resolution transmission electron microscopy imaging have been used to characterize misorientations in graphite spheroids of cast irons. Emphasis is put here on bulk graphite, away from the nucleus as well as from the outer surface of the spheroids in order to get information on their growth during solidification. The results show that spheroidal graphite consists in conical sectors made of elementary blocks piled up on each other. These blocks are elongated along the prismatic a direction of graphite with the c axes roughly parallel to the radius of the spheroids. This implies that the orientation of the blocks rotates around the spheroid centre giving low angle tilting misorientations along tangential direction within each sector. Misorientations between neighbouring sectors are of higher values and their interfaces show rippled layers which are characteristic of defects in graphene. Along a radius of the spheroid, clockwise and anticlockwise twisting between blocks is observed. These observations help challenging some of the models proposed to explain spheroidal growth in cast ions. (paper)

  8. Charged fluid distribution in higher dimensional spheroidal space-time

    Indian Academy of Sciences (India)

    A general solution of Einstein field equations corresponding to a charged fluid distribution on the background of higher dimensional spheroidal space-time is obtained. The solution generates several known solutions for superdense star having spheroidal space-time geometry.

  9. Analytical study of spheroidal dust grains in plasma

    International Nuclear Information System (INIS)

    Zahed, H.; Mahmoodi, J.; Sobhanian, S.

    2006-01-01

    Using the modified spheroidal equations, the potential of a spheroidal conducting grain, floated in a plasma, is calculated. The electric field and capacitance for both prolate and oblate spheroidal grains are investigated. The solutions, obtained up to the second-order approximation, show that the plasma screening causes the equipotential surfaces around the grain to be more elongated or flattened than the potential spheroids of the Laplace equation. This leads to the variation of the plasma concentration around the grain

  10. Dwarf spheroidal galaxies: Keystones of galaxy evolution

    Science.gov (United States)

    Gallagher, John S., III; Wyse, Rosemary F. G.

    1994-01-01

    Dwarf spheroidal galaxies are the most insignificant extragalactic stellar systems in terms of their visibility, but potentially very significant in terms of their role in the formation and evolution of much more luminous galaxies. We discuss the present observational data and their implications for theories of the formation and evolution of both dwarf and giant galaxies. The putative dark-matter content of these low-surface-brightness systems is of particular interest, as is their chemical evolution. Surveys for new dwarf spheroidals hidden behind the stars of our Galaxy and those which are not bound to giant galaxies may give new clues as to the origins of this unique class of galaxy.

  11. A conceptual framework for the evolutionary origins of multicellularity

    International Nuclear Information System (INIS)

    Libby, Eric; B Rainey, Paul

    2013-01-01

    The evolution of multicellular organisms from unicellular counterparts involved a transition in Darwinian individuality from single cells to groups. A particular challenge is to understand the nature of the earliest groups, the causes of their evolution, and the opportunities for emergence of Darwinian properties. Here we outline a conceptual framework based on a logical set of possible pathways for evolution of the simplest self-replicating groups. Central to these pathways is the recognition of a finite number of routes by which genetic information can be transmitted between individual cells and groups. We describe the form and organization of each primordial group state and consider factors affecting persistence and evolution of the nascent multicellular forms. Implications arising from our conceptual framework become apparent when attempting to partition fitness effects at individual and group levels. These are discussed with reference to the evolutionary emergence of individuality and its manifestation in extant multicellular life—including those of marginal Darwinian status. (paper)

  12. Multiscale modeling of the dynamics of multicellular systems

    Science.gov (United States)

    Kosztin, Ioan

    2011-03-01

    Describing the biomechanical properties of cellular systems, regarded as complex highly viscoelastic materials, is a difficult problem of great conceptual and practical value. Here we present a novel approach, referred to as the Cellular Particle Dynamics (CPD) method, for: (i) quantitatively relating biomechanical properties at the cell level to those at the multicellular and tissue level, and (ii) describing and predicting the time evolution of multicellular systems that undergo biomechanical relaxations. In CPD cells are modeled as an ensemble of cellular particles (CPs) that interact via short range contact interactions, characterized by an attractive (adhesive interaction) and a repulsive (excluded volume interaction) component. The time evolution of the spatial conformation of the multicellular system is determined by following the trajectories of all CPs through integration of their equations of motion. Cell and multicellular level biomechanical properties (e.g., viscosity, surface tension and shear modulus) are determined through the combined use of experiments and theory of continuum viscoelastic media. The same biomechanical properties are also ``measured'' computationally by employing the CPD method, the results being expressed in terms of CPD parameters. Once these parameters have been calibrated experimentally, the formalism provides a systematic framework to predict the time evolution of complex multicellular systems during shape-changing biomechanical transformations. By design, the CPD method is rather flexible and most suitable for multiscale modeling of multicellular system. The spatial level of detail of the system can be easily tuned by changing the number of CPs in a cell. Thus, CPD can be used equally well to describe both cell level processes (e.g., the adhesion of two cells) and tissue level processes (e.g., the formation of 3D constructs of millions of cells through bioprinting). Work supported by NSF [FIBR-0526854 and PHY-0957914

  13. Multi-cellular logistics of collective cell migration.

    Directory of Open Access Journals (Sweden)

    Masataka Yamao

    Full Text Available During development, the formation of biological networks (such as organs and neuronal networks is controlled by multicellular transportation phenomena based on cell migration. In multi-cellular systems, cellular locomotion is restricted by physical interactions with other cells in a crowded space, similar to passengers pushing others out of their way on a packed train. The motion of individual cells is intrinsically stochastic and may be viewed as a type of random walk. However, this walk takes place in a noisy environment because the cell interacts with its randomly moving neighbors. Despite this randomness and complexity, development is highly orchestrated and precisely regulated, following genetic (and even epigenetic blueprints. Although individual cell migration has long been studied, the manner in which stochasticity affects multi-cellular transportation within the precisely controlled process of development remains largely unknown. To explore the general principles underlying multicellular migration, we focus on the migration of neural crest cells, which migrate collectively and form streams. We introduce a mechanical model of multi-cellular migration. Simulations based on the model show that the migration mode depends on the relative strengths of the noise from migratory and non-migratory cells. Strong noise from migratory cells and weak noise from surrounding cells causes "collective migration," whereas strong noise from non-migratory cells causes "dispersive migration." Moreover, our theoretical analyses reveal that migratory cells attract each other over long distances, even without direct mechanical contacts. This effective interaction depends on the stochasticity of the migratory and non-migratory cells. On the basis of these findings, we propose that stochastic behavior at the single-cell level works effectively and precisely to achieve collective migration in multi-cellular systems.

  14. Partial discharges in spheroidal voids: Void orientation

    DEFF Research Database (Denmark)

    McAllister, Iain Wilson

    1997-01-01

    Partial discharge transients can be described in terms of the charge induced on the detecting electrode. The influence of the void parameters upon the induced charge is examined and discussed for spheroidal voids. It is shown that a quantitative interpretation of the induced charge requires...

  15. Zero-point energy in spheroidal geometries

    OpenAIRE

    Kitson, A. R.; Signal, A. I.

    2005-01-01

    We study the zero-point energy of a massless scalar field subject to spheroidal boundary conditions. Using the zeta-function method, the zero-point energy is evaluated for small ellipticity. Axially symmetric vector fields are also considered. The results are interpreted within the context of QCD flux tubes and the MIT bag model.

  16. Convection in Slab and Spheroidal Geometries

    Science.gov (United States)

    Porter, David H.; Woodward, Paul R.; Jacobs, Michael L.

    2000-01-01

    Three-dimensional numerical simulations of compressible turbulent thermally driven convection, in both slab and spheroidal geometries, are reviewed and analyzed in terms of velocity spectra and mixing-length theory. The same ideal gas model is used in both geometries, and resulting flows are compared. The piecewise-parabolic method (PPM), with either thermal conductivity or photospheric boundary conditions, is used to solve the fluid equations of motion. Fluid motions in both geometries exhibit a Kolmogorov-like k(sup -5/3) range in their velocity spectra. The longest wavelength modes are energetically dominant in both geometries, typically leading to one convection cell dominating the flow. In spheroidal geometry, a dipolar flow dominates the largest scale convective motions. Downflows are intensely turbulent and up drafts are relatively laminar in both geometries. In slab geometry, correlations between temperature and velocity fluctuations, which lead to the enthalpy flux, are fairly independent of depth. In spheroidal geometry this same correlation increases linearly with radius over the inner 70 percent by radius, in which the local pressure scale heights are a sizable fraction of the radius. The effects from the impenetrable boundary conditions in the slab geometry models are confused with the effects from non-local convection. In spheroidal geometry nonlocal effects, due to coherent plumes, are seen as far as several pressure scale heights from the lower boundary and are clearly distinguishable from boundary effects.

  17. Seamless Combination of Fluorescence-Activated Cell Sorting and Hanging-Drop Networks for Individual Handling and Culturing of Stem Cells and Microtissue Spheroids.

    Science.gov (United States)

    Birchler, Axel; Berger, Mischa; Jäggin, Verena; Lopes, Telma; Etzrodt, Martin; Misun, Patrick Mark; Pena-Francesch, Maria; Schroeder, Timm; Hierlemann, Andreas; Frey, Olivier

    2016-01-19

    Open microfluidic cell culturing devices offer new possibilities to simplify loading, culturing, and harvesting of individual cells or microtissues due to the fact that liquids and cells/microtissues are directly accessible. We present a complete workflow for microfluidic handling and culturing of individual cells and microtissue spheroids, which is based on the hanging-drop network concept: The open microfluidic devices are seamlessly combined with fluorescence-activated cell sorting (FACS), so that individual cells, including stem cells, can be directly sorted into specified culturing compartments in a fully automated way and at high accuracy. Moreover, already assembled microtissue spheroids can be loaded into the microfluidic structures by using a conventional pipet. Cell and microtissue culturing is then performed in hanging drops under controlled perfusion. On-chip drop size control measures were applied to stabilize the system. Cells and microtissue spheroids can be retrieved from the chip by using a parallelized transfer method. The presented methodology holds great promise for combinatorial screening of stem-cell and multicellular-spheroid cultures.

  18. Directed fusion of cardiac spheroids into larger heterocellular microtissues enables investigation of cardiac action potential propagation via cardiac fibroblasts

    Science.gov (United States)

    Markes, Alexander R.; Okundaye, Amenawon O.; Qu, Zhilin; Mende, Ulrike; Choi, Bum-Rak

    2018-01-01

    Multicellular spheroids generated through cellular self-assembly provide cytoarchitectural complexities of native tissue including three-dimensionality, extensive cell-cell contacts, and appropriate cell-extracellular matrix interactions. They are increasingly suggested as building blocks for larger engineered tissues to achieve shapes, organization, heterogeneity, and other biomimetic complexities. Application of these tissue culture platforms is of particular importance in cardiac research as the myocardium is comprised of distinct but intermingled cell types. Here, we generated scaffold-free 3D cardiac microtissue spheroids comprised of cardiac myocytes (CMs) and/or cardiac fibroblasts (CFs) and used them as building blocks to form larger microtissues with different spatial distributions of CMs and CFs. Characterization of fusing homotypic and heterotypic spheroid pairs revealed an important influence of CFs on fusion kinetics, but most strikingly showed rapid fusion kinetics between heterotypic pairs consisting of one CF and one CM spheroid, indicating that CMs and CFs self-sort in vitro into the intermixed morphology found in the healthy myocardium. We then examined electrophysiological integration of fused homotypic and heterotypic microtissues by mapping action potential propagation. Heterocellular elongated microtissues which recapitulate the disproportionate CF spatial distribution seen in the infarcted myocardium showed that action potentials propagate through CF volumes albeit with significant delay. Complementary computational modeling revealed an important role of CF sodium currents and the spatial distribution of the CM-CF boundary in action potential conduction through CF volumes. Taken together, this study provides useful insights for the development of complex, heterocellular engineered 3D tissue constructs and their engraftment via tissue fusion and has implications for arrhythmogenesis in cardiac disease and repair. PMID:29715271

  19. Directed fusion of cardiac spheroids into larger heterocellular microtissues enables investigation of cardiac action potential propagation via cardiac fibroblasts.

    Directory of Open Access Journals (Sweden)

    Tae Yun Kim

    Full Text Available Multicellular spheroids generated through cellular self-assembly provide cytoarchitectural complexities of native tissue including three-dimensionality, extensive cell-cell contacts, and appropriate cell-extracellular matrix interactions. They are increasingly suggested as building blocks for larger engineered tissues to achieve shapes, organization, heterogeneity, and other biomimetic complexities. Application of these tissue culture platforms is of particular importance in cardiac research as the myocardium is comprised of distinct but intermingled cell types. Here, we generated scaffold-free 3D cardiac microtissue spheroids comprised of cardiac myocytes (CMs and/or cardiac fibroblasts (CFs and used them as building blocks to form larger microtissues with different spatial distributions of CMs and CFs. Characterization of fusing homotypic and heterotypic spheroid pairs revealed an important influence of CFs on fusion kinetics, but most strikingly showed rapid fusion kinetics between heterotypic pairs consisting of one CF and one CM spheroid, indicating that CMs and CFs self-sort in vitro into the intermixed morphology found in the healthy myocardium. We then examined electrophysiological integration of fused homotypic and heterotypic microtissues by mapping action potential propagation. Heterocellular elongated microtissues which recapitulate the disproportionate CF spatial distribution seen in the infarcted myocardium showed that action potentials propagate through CF volumes albeit with significant delay. Complementary computational modeling revealed an important role of CF sodium currents and the spatial distribution of the CM-CF boundary in action potential conduction through CF volumes. Taken together, this study provides useful insights for the development of complex, heterocellular engineered 3D tissue constructs and their engraftment via tissue fusion and has implications for arrhythmogenesis in cardiac disease and repair.

  20. A phase field approach for multicellular aggregate fusion in biofabrication.

    Science.gov (United States)

    Yang, Xiaofeng; Sun, Yi; Wang, Qi

    2013-07-01

    We present a modeling and computational approach to study fusion of multicellular aggregates during tissue and organ fabrication, which forms the foundation for the scaffold-less biofabrication of tissues and organs known as bioprinting. It is known as the phase field method, where multicellular aggregates are modeled as mixtures of multiphase complex fluids whose phase mixing or separation is governed by interphase force interactions, mimicking the cell-cell interaction in the multicellular aggregates, and intermediate range interaction mediated by the surrounding hydrogel. The material transport in the mixture is dictated by hydrodynamics as well as forces due to the interphase interactions. In a multicellular aggregate system with fixed number of cells and fixed amount of the hydrogel medium, the effect of cell differentiation, proliferation, and death are neglected in the current model, which can be readily included in the model, and the interaction between different components is dictated by the interaction energy between cell and cell as well as between cell and medium particles, respectively. The modeling approach is applicable to transient simulations of fusion of cellular aggregate systems at the time and length scale appropriate to biofabrication. Numerical experiments are presented to demonstrate fusion and cell sorting during tissue and organ maturation processes in biofabrication.

  1. Multicellular models of intercellular synchronization in circadian neural networks

    International Nuclear Information System (INIS)

    Henson, Michael A.

    2013-01-01

    The circadian clock generates 24 h rhythms that drive physiological and behavioral processes in a diverse range of organisms including microbes, plants, insects, and mammals. Recent experimental advances have produced improved understanding of the molecular mechanisms involved in circadian rhythm generation at the single cell level. However, the intercellular mechanisms that allow large populations of coupled pacemaker cells to synchronize and coordinate their rhythms remain poorly understood. The purpose of this article is to review recent progress in dynamic modeling of the circadian clock with a focus on multicellular models required to describe cell population synchronization. Mammalian systems are emphasized to illustrate the highly heterogeneous structure and rich dynamical behavior of multicellular circadian systems. Available multicellular models are characterized with respect to their single cell descriptions, intercellular coupling mechanisms, and network topologies. Examples drawn from our own research are used to demonstrate the advantages associated with integrating detailed single cell models within realistic multicellular networks for prediction of mammalian system dynamics. Mathematical modeling is shown to represent a powerful tool for understanding the intracellular and intercellular mechanisms utilized to robustly synchronize large populations of highly heterogeneous and sparsely coupled single cell oscillators. The article concludes with some possible directions for future research

  2. Bacterial solutions to multicellularity : a tale of biofilms, filaments and fruiting bodies

    NARCIS (Netherlands)

    Claessen, Dennis; Rozen, Daniel E.; Kuipers, Oscar P.; Søgaard-Andersen, Lotte; Wezel, Gilles P. van

    2014-01-01

    Although bacteria frequently live as unicellular organisms, many spend at least part of their lives in complex communities, and some have adopted truly multicellular lifestyles and have abandoned unicellular growth. These transitions to multicellularity have occurred independently several times for

  3. Using magnetic resonance microscopy to study the growth dynamics of a glioma spheroid in collagen I: A case study

    International Nuclear Information System (INIS)

    Huang, Shuning; Vader, David; Wang, Zhihui; Stemmer-Rachamimov, Anat; Weitz, David A; Dai, Guangping; Rosen, Bruce R; Deisboeck, Thomas S

    2008-01-01

    Highly malignant gliomas are characterized by rapid growth, extensive local tissue infiltration and the resulting overall dismal clinical outcome. Gaining any additional insights into the complex interaction between this aggressive brain tumor and its microenvironment is therefore critical. Currently, the standard imaging modalities to investigate the crucial interface between tumor growth and invasion in vitro are light and confocal laser scanning microscopy. While immensely useful in cell culture, integrating these modalities with this cancer's clinical imaging method of choice, i.e. MRI, is a non-trivial endeavour. However, this integration is necessary, should advanced computational modeling be able to utilize these in vitro data to eventually predict growth behaviour in vivo. We therefore argue that employing the same imaging modality for both the experimental setting and the clinical situation it represents should have significant value from a data integration perspective. In this case study, we have investigated the feasibility of using a specific form of MRI, i.e. magnetic resonance microscopy or MRM, to study the expansion dynamics of a multicellular tumor spheroid in a collagen type I gel. An U87mEGFR human giloblastoma multicellular spheroid (MTS) containing approximately 4·10 3 cells was generated and pipetted into a collagen I gel. The sample was then imaged using a T 2 -weighted 3D spoiled gradient echo pulse sequence on a 14T MRI scanner over a period of 12 hours with a temporal resolution of 3 hours at room temperature. Standard histopathology was performed on the MRM sample, as well as on control samples. We were able to acquire three-dimensional MR images with a spatial resolution of 24 × 24 × 24 μm 3 . Our MRM data successfully documented the volumetric growth dynamics of an MTS in a collagen I gel over the 12-hour period. The histopathology results confirmed cell viability in the MRM sample, yet displayed distinct patterns of cell

  4. Differences in species richness patterns between unicellular and multicellular organisms.

    Science.gov (United States)

    Hillebrand, Helmut; Watermann, Frank; Karez, Rolf; Berninger, Ulrike-G

    2001-01-01

    For unicellular organisms, a lack of effects of local species richness on ecosystem function has been proposed due to their locally high species richness and their ubiquitous distribution. High dispersal ability and high individual numbers may enable unicellular taxa to occur everywhere. Using our own and published data sets on uni- and multicellular organisms, we conducted thorough statistical analyses to test whether (1) unicellular taxa show higher relative local species richness compared to multicellular taxa, (2) unicellular taxa show lower slopes of the species:area relationships and species:individuals relationships, and (3) the species composition of unicellular taxa is less influenced by geographic distance compared to multicellular taxa. We found higher local species richness compared to the global species pool for unicellular organisms than for metazoan taxa. The difference was significant if global species richness was conservatively estimated but not if extrapolated, and therefore higher richness estimates were used. Both microalgae and protozoans showed lower slopes between species richness and sample size (area or individuals) compared to macrozoobenthos, also indicating higher local species richness for unicellular taxa. The similarity of species composition of both benthic diatoms and ciliates decreased with increasing geographic distance. This indicated restricted dispersal ability of protists and the absence of ubiquity. However, a steeper slope between similarity and distance was found for polychaetes and corals, suggesting a stronger effect of distance on the dispersal of metazoans compared to unicellular taxa. In conclusion, we found partly different species richness patterns among uni- and multicellular eukaryotes, but no strict ubiquity of unicellular taxa. Therefore, the effect of local unicellular species richness on ecosystem function has to be reanalyzed. Macroecological patterns suggested for multicellular organisms may differ in

  5. Monitoring the effects of doxorubicin on 3D-spheroid tumor cells in real-time

    Directory of Open Access Journals (Sweden)

    Baek N

    2016-11-01

    halted 3 and 5 days from the start of the treatment in all spheroids. All cell lines maintained their spheroid shape, except SHSY-5, which behaved in an unpredictable manner when exposed to toxic concentrations of DXR. Cytotoxic effects of DXR towards SH-SY5Y seemed to cause degradation of the extracellular matrix, since all cells were dismantled from the spheroid upon cell death. On the other hand, 293T spheroids revealed retarded cellular activity and ATP productions upon DXR treatment throughout the experiment. Since 293T was the embryonic kidney cells, the fast clearance or neutralizations could have made them resistant towards DXR. In conclusion, the same degree of sensitivity from the 2D system did not translate to a 3D culture system, resulting in higher IC50 values than the 2D system. The varying sensitivities and tolerances to drugs could be better understood with a 3D cell culture system. Keywords: doxorubicin, 3D spheroid, cytotoxicity

  6. 3D high-content screening for the identification of compounds that target cells in dormant tumor spheroid regions

    International Nuclear Information System (INIS)

    Wenzel, Carsten; Riefke, Björn; Gründemann, Stephan; Krebs, Alice; Christian, Sven; Prinz, Florian; Osterland, Marc; Golfier, Sven; Räse, Sebastian; Ansari, Nariman; Esner, Milan; Bickle, Marc; Pampaloni, Francesco; Mattheyer, Christian; Stelzer, Ernst H.; Parczyk, Karsten; Prechtl, Stefan; Steigemann, Patrick

    2014-01-01

    Cancer cells in poorly vascularized tumor regions need to adapt to an unfavorable metabolic microenvironment. As distance from supplying blood vessels increases, oxygen and nutrient concentrations decrease and cancer cells react by stopping cell cycle progression and becoming dormant. As cytostatic drugs mainly target proliferating cells, cancer cell dormancy is considered as a major resistance mechanism to this class of anti-cancer drugs. Therefore, substances that target cancer cells in poorly vascularized tumor regions have the potential to enhance cytostatic-based chemotherapy of solid tumors. With three-dimensional growth conditions, multicellular tumor spheroids (MCTS) reproduce several parameters of the tumor microenvironment, including oxygen and nutrient gradients as well as the development of dormant tumor regions. We here report the setup of a 3D cell culture compatible high-content screening system and the identification of nine substances from two commercially available drug libraries that specifically target cells in inner MCTS core regions, while cells in outer MCTS regions or in 2D cell culture remain unaffected. We elucidated the mode of action of the identified compounds as inhibitors of the respiratory chain and show that induction of cell death in inner MCTS core regions critically depends on extracellular glucose concentrations. Finally, combinational treatment with cytostatics showed increased induction of cell death in MCTS. The data presented here shows for the first time a high-content based screening setup on 3D tumor spheroids for the identification of substances that specifically induce cell death in inner tumor spheroid core regions. This validates the approach to use 3D cell culture screening systems to identify substances that would not be detectable by 2D based screening in otherwise similar culture conditions. - Highlights: • Establishment of a novel method for 3D cell culture based high-content screening. • First reported high

  7. 3D high-content screening for the identification of compounds that target cells in dormant tumor spheroid regions

    Energy Technology Data Exchange (ETDEWEB)

    Wenzel, Carsten; Riefke, Björn; Gründemann, Stephan; Krebs, Alice; Christian, Sven; Prinz, Florian; Osterland, Marc; Golfier, Sven; Räse, Sebastian [Bayer Pharma AG, Global Drug Discovery, Muellerstrasse 178, 13353 Berlin (Germany); Ansari, Nariman [Physical Biology Group, Buchmann Institute for Molecular Life Sciences (BMLS), Goethe University Frankfurt (Germany); Esner, Milan; Bickle, Marc [Max Planck Institute of Molecular Cell Biology and Genetics, High-Throughput Technology Development Studio (TDS), Dresden (Germany); Pampaloni, Francesco; Mattheyer, Christian; Stelzer, Ernst H. [Physical Biology Group, Buchmann Institute for Molecular Life Sciences (BMLS), Goethe University Frankfurt (Germany); Parczyk, Karsten; Prechtl, Stefan [Bayer Pharma AG, Global Drug Discovery, Muellerstrasse 178, 13353 Berlin (Germany); Steigemann, Patrick, E-mail: Patrick.Steigemann@bayer.com [Bayer Pharma AG, Global Drug Discovery, Muellerstrasse 178, 13353 Berlin (Germany)

    2014-04-15

    Cancer cells in poorly vascularized tumor regions need to adapt to an unfavorable metabolic microenvironment. As distance from supplying blood vessels increases, oxygen and nutrient concentrations decrease and cancer cells react by stopping cell cycle progression and becoming dormant. As cytostatic drugs mainly target proliferating cells, cancer cell dormancy is considered as a major resistance mechanism to this class of anti-cancer drugs. Therefore, substances that target cancer cells in poorly vascularized tumor regions have the potential to enhance cytostatic-based chemotherapy of solid tumors. With three-dimensional growth conditions, multicellular tumor spheroids (MCTS) reproduce several parameters of the tumor microenvironment, including oxygen and nutrient gradients as well as the development of dormant tumor regions. We here report the setup of a 3D cell culture compatible high-content screening system and the identification of nine substances from two commercially available drug libraries that specifically target cells in inner MCTS core regions, while cells in outer MCTS regions or in 2D cell culture remain unaffected. We elucidated the mode of action of the identified compounds as inhibitors of the respiratory chain and show that induction of cell death in inner MCTS core regions critically depends on extracellular glucose concentrations. Finally, combinational treatment with cytostatics showed increased induction of cell death in MCTS. The data presented here shows for the first time a high-content based screening setup on 3D tumor spheroids for the identification of substances that specifically induce cell death in inner tumor spheroid core regions. This validates the approach to use 3D cell culture screening systems to identify substances that would not be detectable by 2D based screening in otherwise similar culture conditions. - Highlights: • Establishment of a novel method for 3D cell culture based high-content screening. • First reported high

  8. Axisymmetric Eigenmodes of Spheroidal Pure Electron Plasmas

    Science.gov (United States)

    Kawai, Yosuke; Saitoh, Haruhiko; Yoshida, Zensho; Kiwamoto, Yasuhito

    2010-11-01

    The axisymmetric electrostatic eigenmodes of spheroidal pure electron plasmas have been studied experimentally. It is confirmed that the observed spheroidal plasma attains a theoretically expected equilibrium density distribution, with the exception of a low-density halo distribution surrounding the plasma. When the eigenmode frequency observed for the plasma is compared with the frequency predicted by the dispersion relation derived under ideal conditions wherein the temperature is zero and the boundary is located at an infinite distance from the plasma, it is observed that the absolute value of the observed frequency is systematically higher than the theoretical prediction. Experimental examinations and numerical calculations indicate that the upward shift of the eigenmode frequency cannot be accounted for solely by the finite temperature effect, but is significantly affected by image charges induced on the conducting boundary and the resulting distortion of the density profile from the theoretical expectation.

  9. Non-existence of separable spheroidal beams

    International Nuclear Information System (INIS)

    Boyack, Rufus; Lekner, John

    2011-01-01

    We show that ψ = R(ξ)S(η) e imφ , a product of radial and angular oblate spheroidal functions and an azimuthal factor, cannot represent physical free-space scalar beams. The reason lies in the discontinuity in the longitudinal derivative of ψ in the focal plane, where ψ is not a solution of the Helmholtz equation on the disc ξ = 0

  10. Multicellular stalk-like structures in Saccharomyces cerevisiae

    International Nuclear Information System (INIS)

    Engelberg, D.; Mimran, A.; Martinetto, H.; Otto, J.; Simchen, G.; Karin, M.; Fink, G.R.

    1998-01-01

    Stalk formation is a novel pattern of multicellular organization. Yeast cells which survive UV irradiation form colonies that grow vertically to form very long (0.5 to 3.0 cm) and thin (0.5 to 4 mm in diameter) multicellular structures. We describe the conditions required to obtain these stalk-like structures reproducibly in large numbers. Yeast mutants, mutated for control of cell polarity, developmental processes, UV response, and signal transduction cascades were tested and found capable of forming stalk-like structures. We suggest a model that explains the mechanism of stalk formation by mechanical environmental forces. We show that other microorganisms (Candida albicans, Schizosaccharomyces pombe, and Escherichia coli) also form stalks, suggesting that the ability to produce stalks may be a general property of microorganisms. Diploid yeast stalks sporulate at an elevated frequency, raising the possibility that the physiological role of stalks might be disseminating spores

  11. Genome semantics, in silico multicellular systems and the Central Dogma.

    Science.gov (United States)

    Werner, Eric

    2005-03-21

    Genomes with their complexity and size present what appears to be an impossible challenge. Scientists speak in terms of decades or even centuries before we will understand how genomes and their hosts the cell and the city of cells that make up the multicellular context function. We believe that there will be surprisingly quick progress made in our understanding of genomes. The key is to stop taking the Central Dogma as the only direction in which genome research can scale the semantics of genomes. Instead a top-down approach coupled with a bottom-up approach may snare the unwieldy beast and make sense of genomes. The method we propose is to take in silico biology seriously. By developing in silico models of genomes cells and multicellular systems, we position ourselves to develop a theory of meaning for artificial genomes. Then using that develop a natural semantics of genomes.

  12. A multicellular view of cytokinesis in epithelial tissue.

    Science.gov (United States)

    Herszterg, Sophie; Pinheiro, Diana; Bellaïche, Yohanns

    2014-05-01

    The study of cytokinesis in single-cell systems provided a wealth of knowledge on the molecular and biophysical mechanisms controlling daughter cell separation. In this review, we outline recent advances in the understanding of cytokinesis in epithelial tissues. These findings provide evidence for how the cytokinetic machinery adapts to a multicellular context and how the cytokinetic machinery is itself exploited by the tissue for the preservation of tissue function and architecture during proliferation. We propose that cytokinesis in epithelia should be viewed as a multicellular process, whereby the biochemical and mechanical interactions between the dividing cell and its neighbors are essential for successful daughter cell separation while defining epithelial tissue organization and preserving tissue integrity. Copyright © 2013 Elsevier Ltd. All rights reserved.

  13. Acquisition of epithelial-mesenchymal transition and cancer stem-like phenotypes within chitosan-hyaluronan membrane-derived 3D tumor spheroids.

    Science.gov (United States)

    Huang, Yen-Jang; Hsu, Shan-Hui

    2014-12-01

    Cancer drug development has to go through rigorous testing and evaluation processes during pre-clinical in vitro studies. However, the conventional two-dimensional (2D) in vitro culture is often discounted by the insufficiency to present a more typical tumor microenvironment. The multicellular tumor spheroids have been a valuable model to provide more comprehensive assessment of tumor in response to therapeutic strategies. Here, we applied chitosan-hyaluronan (HA) membranes as a platform to promote three-dimensional (3D) tumor spheroid formation. The biological features of tumor spheroids of human non-small cell lung cancer (NSCLC) cells on chitosan-HA membranes were compared to those of 2D cultured cells in vitro. The cells in tumor spheroids cultured on chitosan-HA membranes showed higher levels of stem-like properties and epithelial-mesenchymal transition (EMT) markers, such as NANOG, SOX2, CD44, CD133, N-cadherin, and vimentin, than 2D cultured cells. Moreover, they exhibited enhanced invasive activities and multidrug resistance by the upregulation of MMP2, MMP9, BCRC5, BCL2, MDR1, and ABCG2 as compared with 2D cultured cells. The grafting densities of HA affected the tumor sphere size and mRNA levels of genes on the substrates. These evidences suggest that chitosan-HA membranes may offer a simple and valuable biomaterial platform for rapid generation of tumor spheroids in vitro as well as for further applications in cancer stem cell research and cancer drug screening. Copyright © 2014 Elsevier Ltd. All rights reserved.

  14. Detecting tree-like multicellular life on extrasolar planets.

    Science.gov (United States)

    Doughty, Christopher E; Wolf, Adam

    2010-11-01

    Over the next two decades, NASA and ESA are planning a series of space-based observatories to find Earth-like planets and determine whether life exists on these planets. Previous studies have assessed the likelihood of detecting life through signs of biogenic gases in the atmosphere or a red edge. Biogenic gases and the red edge could be signs of either single-celled or multicellular life. In this study, we propose a technique with which to determine whether tree-like multicellular life exists on extrasolar planets. For multicellular photosynthetic organisms on Earth, competition for light and the need to transport water and nutrients has led to a tree-like body plan characterized by hierarchical branching networks. This design results in a distinct bidirectional reflectance distribution function (BRDF) that causes differing reflectance at different sun/view geometries. BRDF arises from the changing visibility of the shadows cast by objects, and the presence of tree-like structures is clearly distinguishable from flat ground with the same reflectance spectrum. We examined whether the BRDF could detect the existence of tree-like structures on an extrasolar planet by using changes in planetary albedo as a planet orbits its star. We used a semi-empirical BRDF model to simulate vegetation reflectance at different planetary phase angles and both simulated and real cloud cover to calculate disk and rotation-averaged planetary albedo for a vegetated and non-vegetated planet with abundant liquid water. We found that even if the entire planetary albedo were rendered to a single pixel, the rate of increase of albedo as a planet approaches full illumination would be comparatively greater on a vegetated planet than on a non-vegetated planet. Depending on how accurately planetary cloud cover can be resolved and the capabilities of the coronagraph to resolve exoplanets, this technique could theoretically detect tree-like multicellular life on exoplanets in 50 stellar systems.

  15. Morphological and Immunohistochemical Characterization of Canine Osteosarcoma Spheroid Cell Cultures.

    Science.gov (United States)

    Gebhard, C; Gabriel, C; Walter, I

    2016-06-01

    Spheroid cell culture emerges as powerful in vitro tool for experimental tumour research. In this study, we established a scaffold-free three-dimensional spheroid system built from canine osteosarcoma (OS) cells (D17). Spheroids (7, 14 and 19 days of cultivation) and monolayer cultures (2 and 7 days of cultivation) were evaluated and compared on light and electron microscopy. Monolayer and spheroid cultures were tested for vimentin, cytokeratin, alkaline phosphatase, osteocalcin and collagen I by means of immunohistochemistry. The spheroid cell culture exhibited a distinct network of collagen I in particular after 19-day cultivation, whereas in monolayer cultures, collagen I was arranged as a lamellar basal structure. Necrotic centres of large spheroids, as observed in 14- and 19-day cultures, were characterized by significant amounts of osteocalcin. Proliferative activity as determined by Ki-67 immunoreactivity showed an even distribution in two-dimensional cultures. In spheroids, proliferation was predominating in the peripheral areas. Metastasis-associated markers ezrin and S100A4 were shown to be continuously expressed in monolayer and spheroid cultures. We conclude that the scaffold-free spheroid system from canine OS cells has the ability to mimic the architecture of the in vivo tumour, in particular cell-cell and cell-matrix interactions. © 2015 The Authors. Anatomia, Histologia, Embryologia Published by Blackwell Verlag GmbH.

  16. Geometry, packing, and evolutionary paths to increased multicellular size

    Science.gov (United States)

    Jacobeen, Shane; Graba, Elyes C.; Brandys, Colin G.; Day, Thomas C.; Ratcliff, William C.; Yunker, Peter J.

    2018-05-01

    The evolutionary transition to multicellularity transformed life on earth, heralding the evolution of large, complex organisms. Recent experiments demonstrated that laboratory-evolved multicellular "snowflake yeast" readily overcome the physical barriers that limit cluster size by modifying cellular geometry [Jacobeen et al., Nat. Phys. 14, 286 (2018), 10.1038/s41567-017-0002-y]. However, it is unclear why this route to large size is observed, rather than an evolved increase in intercellular bond strength. Here, we use a geometric model of the snowflake yeast growth form to examine the geometric efficiency of increasing size by modifying geometry and bond strength. We find that changing geometry is a far more efficient route to large size than evolving increased intercellular adhesion. In fact, increasing cellular aspect ratio is on average ˜13 times more effective than increasing bond strength at increasing the number of cells in a cluster. Modifying other geometric parameters, such as the geometric arrangement of mother and daughter cells, also had larger effects on cluster size than increasing bond strength. Simulations reveal that as cells reproduce, internal stress in the cluster increases rapidly; thus, increasing bond strength provides diminishing returns in cluster size. Conversely, as cells become more elongated, cellular packing density within the cluster decreases, which substantially decreases the rate of internal stress accumulation. This suggests that geometrically imposed physical constraints may have been a key early selective force guiding the emergence of multicellular complexity.

  17. Cellular packing, mechanical stress and the evolution of multicellularity

    Science.gov (United States)

    Jacobeen, Shane; Pentz, Jennifer T.; Graba, Elyes C.; Brandys, Colin G.; Ratcliff, William C.; Yunker, Peter J.

    2018-03-01

    The evolution of multicellularity set the stage for sustained increases in organismal complexity1-5. However, a fundamental aspect of this transition remains largely unknown: how do simple clusters of cells evolve increased size when confronted by forces capable of breaking intracellular bonds? Here we show that multicellular snowflake yeast clusters6-8 fracture due to crowding-induced mechanical stress. Over seven weeks ( 291 generations) of daily selection for large size, snowflake clusters evolve to increase their radius 1.7-fold by reducing the accumulation of internal stress. During this period, cells within the clusters evolve to be more elongated, concomitant with a decrease in the cellular volume fraction of the clusters. The associated increase in free space reduces the internal stress caused by cellular growth, thus delaying fracture and increasing cluster size. This work demonstrates how readily natural selection finds simple, physical solutions to spatial constraints that limit the evolution of group size—a fundamental step in the evolution of multicellularity.

  18. Global Landslides on Rapidly Spinning Spheroids

    Science.gov (United States)

    Scheeres, Daniel J.; Sanchez, P.

    2013-10-01

    The angle of repose and conditions for global landslides on the surfaces of small, rapidly spinning, spheroidal asteroids are studied. Applying techniques of soil mechanics, we develop a theory for, and examples of, how regolith will fail and flow in this microgravity environment. Our motivation is to develop an understanding of the "top-shaped" class of asteroids based on analytical soil mechanics. Our analysis transforms the entire asteroid surface into a local frame where we can model it as a conventional granular pile with a surface slope, acceleration and height variations as a function of the body's spin rate, shape and density. A general finding is that the lowest point on a rapidly spinning spheroid is at the equator with the effective height of surface material monotonically increasing towards the polar regions, where the height can be larger than the physical radius of the body. We study the failure conditions of both cohesionless and cohesive regolith, and develop specific predictions of the surface profile as a function of the regolith angle of friction and the maximum spin rate experienced by the body. The theory also provides simple guidelines on what the shape may look like, although we do not analyze gravitationally self-consistent evolution of the body shape. The theory is tested with soft-sphere discrete element method granular mechanics simulations to better understand the dynamical aspects of global asteroid landslides. We find significant differences between failure conditions for cohesive and cohesionless regolith. In the case of cohesive regolith, we show that extremely small values of strength (much less than that found in lunar regolith) can stabilize a surface even at very rapid spin rates. Cohesionless surfaces, as expected, fail whenever their surface slopes exceed the angle of friction. Based on our analysis we propose that global landslides and the flow of material towards the equator on spheroidal bodies are precipitated by exogenous

  19. Structure of the Draco Dwarf Spheroidal Galaxy

    OpenAIRE

    Piatek, Slawomir; Pryor, Carlton; Armandroff, Taft E.; Olszewski, Edward W.

    2002-01-01

    This article studies the structure of the Draco dwarf spheroidal galaxy with an emphasis on the question of whether the spatial distribution of its stars has been affected by the tidal interaction with the Milky Way, using R- and V-band CCD photometry for eleven fields. The article reports coordinates for the center, a position angle of the major axis, and the ellipticity. It also reports the results of searches for asymmetries in the structure of Draco. These results, and searches for a ``br...

  20. Process for producing uranium carbide spheroids

    International Nuclear Information System (INIS)

    Shennan, J.V.; Ford, L.H.

    1977-01-01

    The invention deals with a method to fabricate UC spheroids which are filled into moulds made of refractory material for fuel elements. The UC fuel particles are double-coated: a first thin layer of pyrolytic carbon is coated at low temperature 1200-1400 0 C, a record layer of pyrolytic material (e.g. Si c) is coated at a higher temperature (above 1500 0 C) which holds back the fission products. The method is described more closely by means of an example. (GSC) [de

  1. Process for producing uranium carbide spheroids

    International Nuclear Information System (INIS)

    Shennan, J.V.; Ford, L.H.

    1976-01-01

    The invention deals with a method to produce UC spheroids which are filled into molded bodies of fire-proof material for fuel elements. The UC fuel particles are doubly coated: a first thin layer of pyrolytic carbon is coated at low temperature (1,200-1,400 0 C), a second layer of fire-proof material (e.g. SiC) is coated at a higher temperature (above 1,500 0 C) which holds back the fission products. The process is explained in more detail using an example. (GSCH) [de

  2. Short-term effects of radiation in gliolalstoma spheroids

    DEFF Research Database (Denmark)

    Petterson, Stine Asferg; Jakobsen, Ida Pind; Jensen, Stine Skov

    2016-01-01

    was to investigate the short-term effects of radiation of spheroids containing tumor-initiating stem-like cells. We used a patient-derived glioblastoma stem cell enriched culture (T76) and the standard glioblastoma cell line U87. Primary spheroids were irradiated with doses between 2 and 50 Gy and assessed after two...

  3. Cisplatin Resistant Spheroids Model Clinically Relevant Survival Mechanisms in Ovarian Tumors.

    Directory of Open Access Journals (Sweden)

    Winyoo Chowanadisai

    Full Text Available The majority of ovarian tumors eventually recur in a drug resistant form. Using cisplatin sensitive and resistant cell lines assembled into 3D spheroids we profiled gene expression and identified candidate mechanisms and biological pathways associated with cisplatin resistance. OVCAR-8 human ovarian carcinoma cells were exposed to sub-lethal concentrations of cisplatin to create a matched cisplatin-resistant cell line, OVCAR-8R. Genome-wide gene expression profiling of sensitive and resistant ovarian cancer spheroids identified 3,331 significantly differentially expressed probesets coding for 3,139 distinct protein-coding genes (Fc >2, FDR < 0.05 (S2 Table. Despite significant expression changes in some transporters including MDR1, cisplatin resistance was not associated with differences in intracellular cisplatin concentration. Cisplatin resistant cells were significantly enriched for a mesenchymal gene expression signature. OVCAR-8R resistance derived gene sets were significantly more biased to patients with shorter survival. From the most differentially expressed genes, we derived a 17-gene expression signature that identifies ovarian cancer patients with shorter overall survival in three independent datasets. We propose that the use of cisplatin resistant cell lines in 3D spheroid models is a viable approach to gain insight into resistance mechanisms relevant to ovarian tumors in patients. Our data support the emerging concept that ovarian cancers can acquire drug resistance through an epithelial-to-mesenchymal transition.

  4. Variable Stars in the M31 Dwarf Spheroidal Companion Cassiopeia

    Science.gov (United States)

    Pritzl, Barton J.; Armandroff, T. E.; Jacoby, G. H.; Da Costa, G. S.

    2007-12-01

    Dwarf spheroidal galaxies show very diverse star formation histories. For the Galactic dwarf spheroidal galaxies, a correlation exists between Galactocentric distance and the prominence of intermediate-age ( 2 - 10 Gyr) populations. To test whether this correlation exists for the M31 dwarf spheroidal galaxies, we observed the Cassiopeia (And VII) dwarf galaxy, which is one of the most distant M31 dwarf spheroidal galaxies. We will present the results of a variable star search using HST/ACS data, along with a preliminary color-magnitude diagram. From the RR Lyrae stars we can obtain an independent distance and metallicity estimate for the dwarf galaxy. These results will be compared to those found for the other M31 dwarf spheroidal galaxies.This research is supported in part by NASA through grant number GO-11081.11 from the Space Telescope Science Institute.

  5. Development of controlled release spheroids using Buchananiacochinchinesis gum

    Directory of Open Access Journals (Sweden)

    Narayan Babulal Gaikwad

    2013-03-01

    Full Text Available Chirauli nut gum was isolated from the bark of Buchanania cochinchinesis (fam. Anacadiacea and was used as a release modifier for the preparation of Diclofenac sodium spheroids using the extrusion spheronization technique. The process was studied for the effects on variables when making spheroids with satisfactory particle shape, size and size distribution. The prepared spheroids were characterized for surface morphology, qualitative surface porosity, friability, bulk density and flow properties. In vitro studies demonstrated that the release exhibited Fickian diffusion kinetics which was confirmed by the Higuchi and the Korsmeyer-Peppas models. The physico-chemical parameters of the gum could be correlated to the in vitro dissolution profile of the spheroids. The spheroids were not able to sustain the drug releases over 12 hours. A greater concentration of Chirauli nut gum and a process that can accommodate such greater concentrations may produce a formulation capable of significant sustained release.

  6. Design of a Uranium Dioxide Spheroidization System

    Science.gov (United States)

    Cavender, Daniel P.; Mireles, Omar R.; Frendi, Abdelkader

    2013-01-01

    The plasma spheroidization system (PSS) is the first process in the development of tungsten-uranium dioxide (W-UO2) fuel cermets. The PSS process improves particle spherocity and surface morphology for coating by chemical vapor deposition (CVD) process. Angular fully dense particles melt in an argon-hydrogen plasma jet at between 32-36 kW, and become spherical due to surface tension. Surrogate CeO2 powder was used in place of UO2 for system and process parameter development. Particles range in size from 100 - 50 microns in diameter. Student s t-test and hypothesis testing of two proportions statistical methods were applied to characterize and compare the spherocity of pre and post process powders. Particle spherocity was determined by irregularity parameter. Processed powders show great than 800% increase in the number of spherical particles over the stock powder with the mean spherocity only mildly improved. It is recommended that powders be processed two-three times in order to reach the desired spherocity, and that process parameters be optimized for a more narrow particles size range. Keywords: spherocity, spheroidization, plasma, uranium-dioxide, cermet, nuclear, propulsion

  7. Experimental evolution of an alternating uni- and multicellular life cycle in Chlamydomonas reinhardtii

    Science.gov (United States)

    Ratcliff, William C.; Herron, Matthew D.; Howell, Kathryn; Pentz, Jennifer T.; Rosenzweig, Frank; Travisano, Michael

    2013-01-01

    The transition to multicellularity enabled the evolution of large, complex organisms, but early steps in this transition remain poorly understood. Here we show that multicellular complexity, including development from a single cell, can evolve rapidly in a unicellular organism that has never had a multicellular ancestor. We subject the alga Chlamydomonas reinhardtii to conditions that favour multicellularity, resulting in the evolution of a multicellular life cycle in which clusters reproduce via motile unicellular propagules. While a single-cell genetic bottleneck during ontogeny is widely regarded as an adaptation to limit among-cell conflict, its appearance very early in this transition suggests that it did not evolve for this purpose. Instead, we find that unicellular propagules are adaptive even in the absence of intercellular conflict, maximizing cluster-level fecundity. These results demonstrate that the unicellular bottleneck, a trait essential for evolving multicellular complexity, can arise rapidly via co-option of the ancestral unicellular form. PMID:24193369

  8. Energy metabolism and biotransformation as endpoints to pre-screen hepatotoxicity using a liver spheroid model

    International Nuclear Information System (INIS)

    Xu Jinsheng; Purcell, Wendy M.

    2006-01-01

    The current study investigated liver spheroid culture as an in vitro model to evaluate the endpoints relevant to the status of energy metabolism and biotransformation after exposure to test toxicants. Mature rat liver spheroids were exposed to diclofenac, galactosamine, isoniazid, paracetamol, m-dinitrobenzene (m-DNB) and 3-nitroaniline (3-NA) for 24 h. Pyruvate uptake, galactose biotransformation, lactate release and glucose secretion were evaluated after exposure. The results showed that pyruvate uptake and lactate release by mature liver spheroids in culture were maintained at a relatively stable level. These endpoints, together with glucose secretion and galactose biotransformation, were related to and could reflect the status of energy metabolism and biotransformation in hepatocytes. After exposure, all of the test agents significantly reduced glucose secretion, which was shown to be the most sensitive endpoint of those evaluated. Diclofenac, isoniazid, paracetamol and galactosamine reduced lactate release (P < 0.01), but m-DNB increased lactate release (P < 0.01). Diclofenac, isoniazid and paracetamol also reduced pyruvate uptake (P < 0.01), while galactosamine had little discernible effect. Diclofenac, galactosamine, paracetamol and m-DNB also reduced galactose biotransformation (P < 0.01), by contrast, isoniazid did not. The metabolite of m-DNB, 3-NA, which served as a negative control, did not cause significant changes in lactate release, pyruvate uptake or galactose biotransformation. It is concluded that pyruvate uptake, galactose biotransformation, lactate release and glucose secretion can be used as endpoints for evaluating the status of energy metabolism and biotransformation after exposure to test agents using the liver spheroid model to pre-screen hepatotoxicity

  9. Organism Size Promotes the Evolution of Specialized Cells in Multicellular Digital Organisms

    OpenAIRE

    Willensdorfer, Martin

    2007-01-01

    Specialized cells are the essence of complex multicellular life. Fossils allow us to study the modification of specialized, multicellular features such as jaws, scales, and muscular appendages. But it is still unclear what organismal properties contributed to the transition from undifferentiated organisms, which contain only a single cell type, to multicellular organisms with specialized cells. Using digital organisms I study this transition. My simulations show that the transition to special...

  10. 1H MRS can detect dose dependent effects in irradiated tumor cells and spheroids

    International Nuclear Information System (INIS)

    Grande, S.; Viti, V.; Guidoni, L.; Luciani, A.M.

    2003-01-01

    Full text: 1 H MR spectra of tumour cells are often characterised by the presence of intense signals from the methylene fatty acid chains of mobile lipids (ML), mostly triglycerides (TG). In previous work (Magnetic Resonance in Medicine ,1999, 42:248-257), we showed that the intensity of these signals is modulated by cell proliferation. Polyunsaturation levels of the fatty acid chains were also found higher when mobile lipid signals were intense, in agreement with independent findings by other authors (Nat. Med . 1999, 5:1323-1327). Cells from breast carcinoma (MCF 7) were irradiated at increasing doses (5 - 40 Gy) to detect spectral differences in irradiated cells and to relate them to the metabolic breakdown accompanying cell death. Early and late, metabolism mediated , effects were observed. The main effect observed shortly after treatment was a decrease in ML peak intensity, probably from the oxidative damage to the involved lipid structures. On the other hand, when cells were observed after 24, 48 and 72 hours, irradiated cells displayed more intense ML peaks, with a maximum effect after 48 hours, irrespective of the initial intensity of ML signals. The effect was found dose dependent. Also peaks from lipid polyunsaturation were found higher in irradiated samples. Moreover, also phosphorylcholine and glutathione signals were affected by irradiation. Similar effects were found in multicellular spheroids from the same cell strain. Association of the observed changes with apoptotic death is currently under consideration

  11. Cancer across the tree of life: cooperation and cheating in multicellularity

    Science.gov (United States)

    Aktipis, C. Athena; Boddy, Amy M.; Jansen, Gunther; Hibner, Urszula; Hochberg, Michael E.; Maley, Carlo C.; Wilkinson, Gerald S.

    2015-01-01

    Multicellularity is characterized by cooperation among cells for the development, maintenance and reproduction of the multicellular organism. Cancer can be viewed as cheating within this cooperative multicellular system. Complex multicellularity, and the cooperation underlying it, has evolved independently multiple times. We review the existing literature on cancer and cancer-like phenomena across life, not only focusing on complex multicellularity but also reviewing cancer-like phenomena across the tree of life more broadly. We find that cancer is characterized by a breakdown of the central features of cooperation that characterize multicellularity, including cheating in proliferation inhibition, cell death, division of labour, resource allocation and extracellular environment maintenance (which we term the five foundations of multicellularity). Cheating on division of labour, exhibited by a lack of differentiation and disorganized cell masses, has been observed in all forms of multicellularity. This suggests that deregulation of differentiation is a fundamental and universal aspect of carcinogenesis that may be underappreciated in cancer biology. Understanding cancer as a breakdown of multicellular cooperation provides novel insights into cancer hallmarks and suggests a set of assays and biomarkers that can be applied across species and characterize the fundamental requirements for generating a cancer. PMID:26056363

  12. Differentiation of human mesenchymal stem cell spheroids under microgravity conditions

    Directory of Open Access Journals (Sweden)

    Wolfgang H Cerwinka

    2012-01-01

    Full Text Available To develop and characterize a novel cell culture method for the generation of undifferentiated and differentiated human mesenchymal stem cell 3D structures, we utilized the RWV system with a gelatin-based scaffold. 3 × 106 cells generated homogeneous spheroids and maximum spheroid loading was accomplished after 3 days of culture. Spheroids cultured in undifferentiated spheroids of 3 and 10 days retained expression of CD44, without expression of differentiation markers. Spheroids cultured in adipogenic and osteogenic differentiation media exhibited oil red O staining and von Kossa staining, respectively. Further characterization of osteogenic lineage, showed that 10 day spheroids exhibited stronger calcification than any other experimental group corresponding with significant expression of vitamin D receptor, alkaline phosphatase, and ERp60 . In conclusion this study describes a novel RWV culture method that allowed efficacious engineering of undifferentiated human mesenchymal stem cell spheroids and rapid osteogenic differentiation. The use of gelatin scaffolds holds promise to design implantable stem cell tissue of various sizes and shapes for future regenerative treatment.

  13. Ellipticity behaviour of relativistic Maclaurin spheroids

    International Nuclear Information System (INIS)

    Abramowicz, M.A.; Miller, J.C.; International School of Advanced Studies, Trieste; Oxford Univ.

    1990-01-01

    Chandrasekhar and Miller (1974 Mon. Not. R. Astr. Soc., 167, 63) found that, according to the general theory of relativity, when slowly rotating spheroids contract, conserving angular momentum, their eccentricity does not increase monotonically, as is the case for Newtonian gravity, but instead reaches a maximum (when the mean radius is a few times the Schwarzschild radius) and then decreases again. We show here that this can be explained within the framework of the recent discussion of rotational effects in a strong gravitational field. We suggest that it can be convenient to introduce a 'corrected' Newtonian theory, incorporating terms to approximate strong field effects, and that such a corrected theory could be useful for numerical studies. (author)

  14. Electron, proton, neutron as spheroidical particles

    International Nuclear Information System (INIS)

    Bagge, E.R.

    1993-01-01

    It is shown that it is possible to describe the electron and the proton at rest within the framework of Dirac's relativistic theory of particles as electro-magnetic stable, spheroidal particles like balloons with very thin envelopes. Their properties, especially their spins and their magnetic momenta, are exactly those, which have been measured at first and later on derived by Dirac. In this picture the neutron plays the role of a system of two concentric and synchronically rotating balloons with a small distance between them at a positive energetic minimum of balance at 1.26 MeV. The magnetic moment of this particle has a negative sign and is of the correct size. (orig.)

  15. Plasma spheroidizing and cladding of powders

    International Nuclear Information System (INIS)

    Petrunichev, V.A.; Averin, V.V.; Sorokin, L.M.; Koroleva, E.B.

    1987-01-01

    Arc and high-frequency plasmatrons are used for spheroidizing nickel and chromium-base alloy particles. Different plasma-forming medium compositions are used in the arc variant and the effect of these media on the plasma treatment product is demonstrated. For a high-frequency plasmatron, a long time of plasma contact with the powder leads to the transfer of the part of the material from the treated particles into vaporous state with subsequent condensation at the outlet from the discharge zone. Results of investigations into the formation of metal coatings on oxide and carbide particles during plasma-arc action are also presented. Representative data on the output of particles with coating are obtained and factors, providing for the optimal particle cladding conditions, are indicated

  16. Magnetohydrodynamic equilibrium with spheroidal plasma-vacuum interface

    International Nuclear Information System (INIS)

    Kaneko, Shobu; Chiyoda, Katsuji; Hirota, Isao.

    1983-01-01

    The Grad-Shafranov equations for an oblate and a prolate spheroidal plasmas are solved analytically under the assumptions, Bsub(phi) = 0 and dp/dpsi = constant. Here Bsub(phi) is the toroidal magnetic field, p is the kinetic pressure, and psi is the magnetic flux function. The plasmas in magnetohydrodynamic equilibrium are shown to be toroidal. The equilibrium magnetic-field configurations outside the spheroidal plasmas are considerably different from that of a spherical plasma. A line cusp or two point cusps appear outside the oblate or the prolate spheroidal plasma, respectively. (author)

  17. [Reparative and neoplastic spheroid cellular structures and their mathematical model].

    Science.gov (United States)

    Kogan, E A; Namiot, V A; Demura, T A; Faĭzullina, N M; Sukhikh, G T

    2014-01-01

    Spheroid cell structures in the cell cultures have been described and are used for studying cell-cell and cell- matrix interactions. At the same time, spheroid cell structure participation in the repair and development of cancer in vivo remains unexplored. The aim of this study was to investigate the cellular composition of spherical structures and their functional significance in the repair of squamous epithelium in human papilloma virus-associated cervical pathology--chronic cervicitis and cervical intraepithelial neoplasia 1-3 degree, and also construct a mathematical model to explain the development and behavior of such spheroid cell structure.

  18. Powder processing and spheroidizing with thermal inductively coupled plasma

    International Nuclear Information System (INIS)

    Nutsch, G.; Linke, P.; Zakharian, S.; Dzur, B.; Weiss, K.-H.

    2001-01-01

    Processing of advanced powder materials for the spraying industry is one of the most promising applications of the thermal RF inductively coupled plasma. By selecting the feedstock carefully and adjusting the RF plasma parameters, unique materials with high quality can be achieved. Powders injected in the hot plasma core emerge with modified shapes, morphology, crystal structure and chemical composition. Ceramic oxide powders such as Al 2 O 3 , ZrO 2 , SiO 2 are spheroidized with a high spheroidization rate. By using the RF induction plasma spheroidizing process tungsten melt carbide powders are obtained with a high spheroidization rate at high feeding rates by densification of agglomerated powders consisting of di-tungsten carbide and monocarbide with a definite composition. This kind of ball-like powders is particularly suited for wear resistant applications. (author)

  19. Spheroidization of transition metal carbides in low temperature plasma

    International Nuclear Information System (INIS)

    Klinskaya, N.A.; Koroleva, E.B.; Petrunichev, V.A.; Rybalko, O.F.; Solov'ev, P.V.; Ugol'nikova, T.A.

    1986-01-01

    Plasma process of preparation of titanium, tungsten and chromium carbide spherical powders with the main particle size 40-80 μm is considered. Spheroidization degree, granulometric and phase composition of the product are investigated

  20. Green Algae and the Origins of Multicellularity in the Plant Kingdom

    Science.gov (United States)

    Umen, James G.

    2014-01-01

    The green lineage of chlorophyte algae and streptophytes form a large and diverse clade with multiple independent transitions to produce multicellular and/or macroscopically complex organization. In this review, I focus on two of the best-studied multicellular groups of green algae: charophytes and volvocines. Charophyte algae are the closest relatives of land plants and encompass the transition from unicellularity to simple multicellularity. Many of the innovations present in land plants have their roots in the cell and developmental biology of charophyte algae. Volvocine algae evolved an independent route to multicellularity that is captured by a graded series of increasing cell-type specialization and developmental complexity. The study of volvocine algae has provided unprecedented insights into the innovations required to achieve multicellularity. PMID:25324214

  1. Reliability analysis of multicellular system architectures for low-cost satellites

    Science.gov (United States)

    Erlank, A. O.; Bridges, C. P.

    2018-06-01

    Multicellular system architectures are proposed as a solution to the problem of low reliability currently seen amongst small, low cost satellites. In a multicellular architecture, a set of independent k-out-of-n systems mimic the cells of a biological organism. In order to be beneficial, a multicellular architecture must provide more reliability per unit of overhead than traditional forms of redundancy. The overheads include power consumption, volume and mass. This paper describes the derivation of an analytical model for predicting a multicellular system's lifetime. The performance of such architectures is compared against that of several common forms of redundancy and proven to be beneficial under certain circumstances. In addition, the problem of peripheral interfaces and cross-strapping is investigated using a purpose-developed, multicellular simulation environment. Finally, two case studies are presented based on a prototype cell implementation, which demonstrate the feasibility of the proposed architecture.

  2. Fragile DNA Repair Mechanism Reduces Ageing in Multicellular Model

    DEFF Research Database (Denmark)

    Bendtsen, Kristian Moss; Juul, Jeppe Søgaard; Trusina, Ala

    2012-01-01

    increases the amount of unrepaired DNA damage. Despite this vicious circle, we ask, can cells maintain a high DNA repair capacity for some time or is repair capacity bound to continuously decline with age? We here present a simple mathematical model for ageing in multicellular systems where cells subjected...... to DNA damage can undergo full repair, go apoptotic, or accumulate mutations thus reducing DNA repair capacity. Our model predicts that at the tissue level repair rate does not continuously decline with age, but instead has a characteristic extended period of high and non-declining DNA repair capacity......DNA damages, as well as mutations, increase with age. It is believed that these result from increased genotoxic stress and decreased capacity for DNA repair. The two causes are not independent, DNA damage can, for example, through mutations, compromise the capacity for DNA repair, which in turn...

  3. Thinking about Bacillus subtilis as a multicellular organism.

    Science.gov (United States)

    Aguilar, Claudio; Vlamakis, Hera; Losick, Richard; Kolter, Roberto

    2007-12-01

    Initial attempts to use colony morphogenesis as a tool to investigate bacterial multicellularity were limited by the fact that laboratory strains often have lost many of their developmental properties. Recent advances in elucidating the molecular mechanisms underlying colony morphogenesis have been made possible through the use of undomesticated strains. In particular, Bacillus subtilis has proven to be a remarkable model system to study colony morphogenesis because of its well-characterized developmental features. Genetic screens that analyze mutants defective in colony morphology have led to the discovery of an intricate regulatory network that controls the production of an extracellular matrix. This matrix is essential for the development of complex colony architecture characterized by aerial projections that serve as preferential sites for sporulation. While much progress has been made, the challenge for future studies will be to determine the underlying mechanisms that regulate development such that differentiation occurs in a spatially and temporally organized manner.

  4. Dwarf Spheroidal Satellite Formation in a Reionized Local Group

    OpenAIRE

    Milosavljevic, Milos; Bromm, Volker

    2013-01-01

    Dwarf spheroidal satellite galaxies have emerged a powerful probe of small-scale dark matter clustering and of cosmic reionization. They exhibit structural and chemical continuity with dwarf irregular galaxies in the field and with spheroidal galaxies in high-density environments. By combining empirical constraints derived for star formation at low gas column densities and metallicities in the local universe with a model for dark matter and baryonic mass assembly, we provide an analytical des...

  5. Correlation between grade of pearlite spheroidization and laser induced spectra

    Science.gov (United States)

    Yao, Shunchun; Dong, Meirong; Lu, Jidong; Li, Jun; Dong, Xuan

    2013-12-01

    Laser induced breakdown spectroscopy (LIBS) which is used traditionally as a spectrochemical analytical technique was employed to analyze the grade of pearlite spheroidization. Three 12Cr1MoV steel specimens with different grades of pearlite spheroidization were ablated to produce plasma by pulse laser at 266 nm. In order to determine the optimal temporal condition and plasma parameters for correlating the grade of pearlite spheroidization and laser induced spectra, a set of spectra at different delays were analyzed by the principal component analysis method. Then, the relationship between plasma temperature, intensity ratios of ionic to atomic lines and grade of pearlite spheroidization was studied. The analysis results show that the laser induced spectra of different grades of pearlite spheroidization can be readily identifiable by principal component analysis in the range of 271.941-289.672 nm with 1000 ns delay time. It is also found that a good agreement exists between the Fe ionic to atomic line ratios and the tensile strength, whereas there is no obvious difference in the plasma temperature. Therefore, LIBS may be applied not only as a spectrochemical analytical technique but also as a new way to estimate the grade of pearlite spheroidization.

  6. Correlation between grade of pearlite spheroidization and laser induced spectra

    International Nuclear Information System (INIS)

    Yao, Shunchun; Dong, Meirong; Lu, Jidong; Li, Jun; Dong, Xuan

    2013-01-01

    Laser induced breakdown spectroscopy (LIBS) which is used traditionally as a spectrochemical analytical technique was employed to analyze the grade of pearlite spheroidization. Three 12Cr1MoV steel specimens with different grades of pearlite spheroidization were ablated to produce plasma by pulse laser at 266 nm. In order to determine the optimal temporal condition and plasma parameters for correlating the grade of pearlite spheroidization and laser induced spectra, a set of spectra at different delays were analyzed by the principal component analysis method. Then, the relationship between plasma temperature, intensity ratios of ionic to atomic lines and grade of pearlite spheroidization was studied. The analysis results show that the laser induced spectra of different grades of pearlite spheroidization can be readily identifiable by principal component analysis in the range of 271.941–289.672 nm with 1000 ns delay time. It is also found that a good agreement exists between the Fe ionic to atomic line ratios and the tensile strength, whereas there is no obvious difference in the plasma temperature. Therefore, LIBS may be applied not only as a spectrochemical analytical technique but also as a new way to estimate the grade of pearlite spheroidization. (paper)

  7. Implications of the dwarfs spheroidal galaxy mass-metallicity relation

    International Nuclear Information System (INIS)

    Smith, G.H.

    1985-01-01

    The properties of the mass-metallicity relation among dwarf spheroidal galaxies are discussed in terms of a model which assumes that the internal chemical evolution of the dwarf spheroidals was promoted by supernova activity. The model can be used to explain the observed dwarf spheroidal mass-metallicity relation assuming the present mass of these systems M sub s is proportional to their initial masses M as M sub s varies according to a power-law index of exp 7/4. It is inferred from the power-law dependence of M on the proto-cloud radius that the most massive dwarf spheroids were formed from the densest clouds. The observed slope of the mass-metallicity relation for dwarf spheroidal galaxies is found to be significantly different from theoretical estimates of this slope for elliptical galaxies. It is suggested that the difference may imply that spheroidal dwarfs and elliptical galaxies had different formation histories, confirming Kormendy's (1985) observations of differences in the brightness and luminosity trends. 54 references

  8. Minimal resin embedding of multicellular specimens for targeted FIB-SEM imaging.

    Science.gov (United States)

    Schieber, Nicole L; Machado, Pedro; Markert, Sebastian M; Stigloher, Christian; Schwab, Yannick; Steyer, Anna M

    2017-01-01

    Correlative light and electron microscopy (CLEM) is a powerful tool to perform ultrastructural analysis of targeted tissues or cells. The large field of view of the light microscope (LM) enables quick and efficient surveys of the whole specimen. It is also compatible with live imaging, giving access to functional assays. CLEM protocols take advantage of the features to efficiently retrace the position of targeted sites when switching from one modality to the other. They more often rely on anatomical cues that are visible both by light and electron microscopy. We present here a simple workflow where multicellular specimens are embedded in minimal amounts of resin, exposing their surface topology that can be imaged by scanning electron microscopy (SEM). LM and SEM both benefit from a large field of view that can cover whole model organisms. As a result, targeting specific anatomic locations by focused ion beam-SEM (FIB-SEM) tomography becomes straightforward. We illustrate this application on three different model organisms, used in our laboratory: the zebrafish embryo Danio rerio, the marine worm Platynereis dumerilii, and the dauer larva of the nematode Caenorhabditis elegans. Here we focus on the experimental steps to reduce the amount of resin covering the samples and to image the specimens inside an FIB-SEM. We expect this approach to have widespread applications for volume electron microscopy on multiple model organisms. Copyright © 2017 Elsevier Inc. All rights reserved.

  9. Phylostratigraphic tracking of cancer genes suggests a link to the emergence of multicellularity in metazoa

    Directory of Open Access Journals (Sweden)

    Domazet-Lošo Tomislav

    2010-05-01

    Full Text Available Abstract Background Phylostratigraphy is a method used to correlate the evolutionary origin of founder genes (that is, functional founder protein domains of gene families with particular macroevolutionary transitions. It is based on a model of genome evolution that suggests that the origin of complex phenotypic innovations will be accompanied by the emergence of such founder genes, the descendants of which can still be traced in extant organisms. The origin of multicellularity can be considered to be a macroevolutionary transition, for which new gene functions would have been required. Cancer should be tightly connected to multicellular life since it can be viewed as a malfunction of interaction between cells in a multicellular organism. A phylostratigraphic tracking of the origin of cancer genes should, therefore, also provide insights into the origin of multicellularity. Results We find two strong peaks of the emergence of cancer related protein domains, one at the time of the origin of the first cell and the other around the time of the evolution of the multicellular metazoan organisms. These peaks correlate with two major classes of cancer genes, the 'caretakers', which are involved in general functions that support genome stability and the 'gatekeepers', which are involved in cellular signalling and growth processes. Interestingly, this phylogenetic succession mirrors the ontogenetic succession of tumour progression, where mutations in caretakers are thought to precede mutations in gatekeepers. Conclusions A link between multicellularity and formation of cancer has often been predicted. However, this has not so far been explicitly tested. Although we find that a significant number of protein domains involved in cancer predate the origin of multicellularity, the second peak of cancer protein domain emergence is, indeed, connected to a phylogenetic level where multicellular animals have emerged. The fact that we can find a strong and

  10. Development of lacrimal gland spheroids for lacrimal gland tissue regeneration.

    Science.gov (United States)

    Massie, Isobel; Spaniol, Kristina; Barbian, Andreas; Geerling, Gerd; Metzger, Marco; Schrader, Stefan

    2018-04-01

    Severe dry eye syndrome resulting from lacrimal gland (LG) dysfunction can cause blindness, yet treatments remain palliative. In vitro reconstruction of LG tissue could provide a curative treatment. We aimed to combine epithelial cells with endothelial cells and mesenchymal stem cells (MSCs) to form a 3D functional unit. Epithelial cells and MSCs were isolated from porcine LG; endothelial cells were isolated from human foreskin. MSCs were characterised (flow cytometry and differentiation potential assays). All 3 cell types were combined on Matrigel and spheroid formation observed. Spheroids were characterised [immunohistochemistry (IHC) and transmission electron microscopy] and function assessed (β-hexosaminidase assay). Spheroids were transferred to decellularised jejunum (SIS-Muc) in dynamic cultures for 1 week before further characterisation. MSCs did not express CD31 but expressed CD44 and CD105 and differentiated towards osteogenic and adipogenic lineages. Spheroids formed on Matrigel within 18 hr, contracting to ~10% of the well area (p function was increased in spheroids cf. monolayer controls (p function (p < .05), viability (p < .05), and proliferation decreased, whilst apoptosis increased. On SIS-Muc under dynamic culture, however, spheroids continued to proliferate to repopulate SIS-Muc. IHC revealed LG epithelial cells coexpressing pan-cytokeratin and lysozyme, as well as endothelial cells and MSCs and cells remained capable of responding to carbachol (p < .05). These spheroids could form the basis of a regenerative medicine treatment approach for dry eye syndrome. In vivo studies are required to evaluate this further. Copyright © 2017 John Wiley & Sons, Ltd.

  11. Human Cardiac Progenitor Spheroids Exhibit Enhanced Engraftment Potential.

    Directory of Open Access Journals (Sweden)

    Francesca Oltolina

    Full Text Available A major obstacle to an effective myocardium stem cell therapy has always been the delivery and survival of implanted stem cells in the heart. Better engraftment can be achieved if cells are administered as cell aggregates, which maintain their extra-cellular matrix (ECM. We have generated spheroid aggregates in less than 24 h by seeding human cardiac progenitor cells (hCPCs onto methylcellulose hydrogel-coated microwells. Cells within spheroids maintained the expression of stemness/mesenchymal and ECM markers, growth factors and their cognate receptors, cardiac commitment factors, and metalloproteases, as detected by immunofluorescence, q-RT-PCR and immunoarray, and expressed a higher, but regulated, telomerase activity. Compared to cells in monolayers, 3D spheroids secreted also bFGF and showed MMP2 activity. When spheroids were seeded on culture plates, the cells quickly migrated, displaying an increased wound healing ability with or without pharmacological modulation, and reached confluence at a higher rate than cells from conventional monolayers. When spheroids were injected in the heart wall of healthy mice, some cells migrated from the spheroids, engrafted, and remained detectable for at least 1 week after transplantation, while, when the same amount of cells was injected as suspension, no cells were detectable three days after injection. Cells from spheroids displayed the same engraftment capability when they were injected in cardiotoxin-injured myocardium. Our study shows that spherical in vivo ready-to-implant scaffold-less aggregates of hCPCs able to engraft also in the hostile environment of an injured myocardium can be produced with an economic, easy and fast protocol.

  12. Coulomb energy of uniformly charged spheroidal shell systems.

    Science.gov (United States)

    Jadhao, Vikram; Yao, Zhenwei; Thomas, Creighton K; de la Cruz, Monica Olvera

    2015-03-01

    We provide exact expressions for the electrostatic energy of uniformly charged prolate and oblate spheroidal shells. We find that uniformly charged prolate spheroids of eccentricity greater than 0.9 have lower Coulomb energy than a sphere of the same area. For the volume-constrained case, we find that a sphere has the highest Coulomb energy among all spheroidal shells. Further, we derive the change in the Coulomb energy of a uniformly charged shell due to small, area-conserving perturbations on the spherical shape. Our perturbation calculations show that buckling-type deformations on a sphere can lower the Coulomb energy. Finally, we consider the possibility of counterion condensation on the spheroidal shell surface. We employ a Manning-Oosawa two-state model approximation to evaluate the renormalized charge and analyze the behavior of the equilibrium free energy as a function of the shell's aspect ratio for both area-constrained and volume-constrained cases. Counterion condensation is seen to favor the formation of spheroidal structures over a sphere of equal area for high values of shell volume fractions.

  13. Biocellion: accelerating computer simulation of multicellular biological system models.

    Science.gov (United States)

    Kang, Seunghwa; Kahan, Simon; McDermott, Jason; Flann, Nicholas; Shmulevich, Ilya

    2014-11-01

    Biological system behaviors are often the outcome of complex interactions among a large number of cells and their biotic and abiotic environment. Computational biologists attempt to understand, predict and manipulate biological system behavior through mathematical modeling and computer simulation. Discrete agent-based modeling (in combination with high-resolution grids to model the extracellular environment) is a popular approach for building biological system models. However, the computational complexity of this approach forces computational biologists to resort to coarser resolution approaches to simulate large biological systems. High-performance parallel computers have the potential to address the computing challenge, but writing efficient software for parallel computers is difficult and time-consuming. We have developed Biocellion, a high-performance software framework, to solve this computing challenge using parallel computers. To support a wide range of multicellular biological system models, Biocellion asks users to provide their model specifics by filling the function body of pre-defined model routines. Using Biocellion, modelers without parallel computing expertise can efficiently exploit parallel computers with less effort than writing sequential programs from scratch. We simulate cell sorting, microbial patterning and a bacterial system in soil aggregate as case studies. Biocellion runs on x86 compatible systems with the 64 bit Linux operating system and is freely available for academic use. Visit http://biocellion.com for additional information. © The Author 2014. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

  14. Integrated biocircuits: engineering functional multicellular circuits and devices

    Science.gov (United States)

    Prox, Jordan; Smith, Tory; Holl, Chad; Chehade, Nick; Guo, Liang

    2018-04-01

    Objective. Implantable neurotechnologies have revolutionized neuromodulatory medicine for treating the dysfunction of diseased neural circuitry. However, challenges with biocompatibility and lack of full control over neural network communication and function limits the potential to create more stable and robust neuromodulation devices. Thus, we propose a platform technology of implantable and programmable cellular systems, namely Integrated Biocircuits, which use only cells as the functional components of the device. Approach. We envision the foundational principles for this concept begins with novel in vitro platforms used for the study and reconstruction of cellular circuitry. Additionally, recent advancements in organoid and 3D culture systems account for microenvironment factors of cytoarchitecture to construct multicellular circuits as they are normally formed in the brain. We explore the current state of the art of these platforms to provide knowledge of their advancements in circuit fabrication and identify the current biological principles that could be applied in designing integrated biocircuit devices. Main results. We have highlighted the exemplary methodologies and techniques of in vitro circuit fabrication and propose the integration of selected controllable parameters, which would be required in creating suitable biodevices. Significance. We provide our perspective and propose new insights into the future of neuromodulaion devices within the scope of living cellular systems that can be applied in designing more reliable and biocompatible stimulation-based neuroprosthetics.

  15. Differential superiority of heavy charged-particle irradiation to x-rays: Studies on biological effectivenes and side effect mechanisms in multicellular tumor and normal tissue models

    Directory of Open Access Journals (Sweden)

    Stefan eWalenta

    2016-02-01

    Full Text Available This review is focused on the radiobiology of carbon ions compared to x-rays using multicellular models of tumors and normal mucosa. The first part summarizes basic radiobiological effects, as observed in cancer cells. The second, more clinically oriented part of the review deals with radiation-induced cell migration and mucositis.Multicellular spheroids (MCS from V79 hamster cells were irradiated with x-rays or carbon ions under ambient or restricted oxygen supply conditions. Oxygen enhancement ratios (OER were 2.9, 2.8, and 1.4 for irradiation with photons, 12C+6 in the plateau region, and 12C+6 in the Bragg peak, respectively. A relative biological effectiveness (RBE of 4.3 and 2.1 for ambient pO2 and hypoxia was obtained, respectively. The high effectiveness of carbon ions was reflected by an enhanced accumulation of cells in G2/M, and a dose-dependent massive induction of apoptosis. Clinically relevant doses (3 Gy of x-rays induced an increase in migratory activity of U87 but not of LN229 or HCT116 tumor cells. Such an increase in cell motility following irradiation in situ could be the source of recurrence. In contrast, carbon ion treatment was associated with a dose-dependent decrease in migration with all cell lines and under all conditions investigated. The radiation-induced loss of cell motility was correlated, in most cases, with corresponding changes in 1 integrin expression. Unlike with particles, the photon-induced increase in cell migration was paralleled by an elevated phosphorylation status of the epidermal growth factor receptor (EGFR and AKT-ERK1/2 pathway. Comparing the gene toxicity of x-rays with that of particles using the gamma-H2AX technique in organotypic cultures of the oral mucosa, the superior effectiveness of heavy ions was confirmed by a two-fold higher number of foci per nucleus. Pro-inflammatory signs, however, were similar for both treatment modalities, e. g., the activation of NFkappaB, and the release of IL

  16. Observation of multicellular spinning behavior of Proteus mirabilis by atomic force microscopy and multifunctional microscopy.

    Science.gov (United States)

    Liu, Yanxia; Deng, Yuanxin; Luo, Shuxiu; Deng, Yu; Guo, Linming; Xu, Weiwei; Liu, Lei; Liu, Junkang

    2014-01-01

    This study aimed to observe the multicellular spinning behavior of Proteus mirabilis by atomic force microscopy (AFM) and multifunctional microscopy in order to understand the mechanism underlying this spinning movement and its biological significance. Multifunctional microscopy with charge-coupled device (CCD) and real-time AFM showed changes in cell structure and shape of P. mirabilis during multicellular spinning movement. Specifically, the morphological characteristics of P. mirabilis, multicellular spinning dynamics, and unique movement were observed. Our findings indicate that the multicellular spinning behavior of P. mirabilis may be used to collect nutrients, perform colonization, and squeeze out competitors. The movement characteristics of P. mirabilis are vital to the organism's biological adaptability to the surrounding environment. Copyright © 2013 Elsevier Ltd. All rights reserved.

  17. Cell proliferation kinetics and radiation response in 9L tumor spheroids

    Energy Technology Data Exchange (ETDEWEB)

    Sweigert, S.E.

    1984-05-01

    Cell kinetic parameters, including population doubling-time, cell cycle time, and growth fraction, were measured in 9L gliosarcoma spheroids. These parameters were studied as the spheroids grew from 50 ..mu..m to over 900 ..mu..m in diameter. Experiments relating the cell kinetic parameters to the radiation response of 9L spheroids were also carried out. The major findings were that the average cell cycle time (T/sub c/), is considerably longer in large spheroids than in exponentially-growing monolayers, the radiosensitivity of noncycling (but still viable) cells in spheroids is not significantly different from that of cycling spheroid cells, and the radiation-induced division delay is approximately twice as long in spheroid cells as in monolayer cells given equal radiation doses. The cell loss factor for spheroids of various sizes was calculated, by using the measured kinetic parameters in the basic equations for growth of a cell population. 157 references, 6 figures, 3 tables.

  18. Cell proliferation kinetics and radiation response in 9L tumor spheroids

    International Nuclear Information System (INIS)

    Sweigert, S.E.

    1984-05-01

    Cell kinetic parameters, including population doubling-time, cell cycle time, and growth fraction, were measured in 9L gliosarcoma spheroids. These parameters were studied as the spheroids grew from 50 μm to over 900 μm in diameter. Experiments relating the cell kinetic parameters to the radiation response of 9L spheroids were also carried out. The major findings were that the average cell cycle time (T/sub c/), is considerably longer in large spheroids than in exponentially-growing monolayers, the radiosensitivity of noncycling (but still viable) cells in spheroids is not significantly different from that of cycling spheroid cells, and the radiation-induced division delay is approximately twice as long in spheroid cells as in monolayer cells given equal radiation doses. The cell loss factor for spheroids of various sizes was calculated, by using the measured kinetic parameters in the basic equations for growth of a cell population. 157 references, 6 figures, 3 tables

  19. Process for titanium powders spheroidization by RF induction plasma

    International Nuclear Information System (INIS)

    Gu Zhongtao; Ye Gaoying; Liu Chuandong; Tong Honghui

    2010-01-01

    Spherical titanium (Ti) particles were obtained by the process of heating irregularly shaped Ti powders under the radio frequency induction plasma (RF induction plasma) condition. The effect of feed rate, various dispersion methods and Ti particle size on the spheroidization efficiency was studied. The efficiency of the spheroidization is evaluated through the measurements of the percentage of powder spheroidized based on the electron microscopic observations and the tap density measurement of the processed powder. During the short flight of the particles in the plasma flow, of the order of a few milliseconds, the individual titanium particles of the powder are heated and melt, forming a spherical liquid droplet which upon freezing gives rise to the formation of a perfectly dense spherical solid particle. So RF induction plasma is a promising method for the preparation of spherical titanium powders with high flow ability. (authors)

  20. Spheroidization of glass powders for glass ionomer cements.

    Science.gov (United States)

    Gu, Y W; Yap, A U J; Cheang, P; Kumar, R

    2004-08-01

    Commercial angular glass powders were spheroidized using both the flame spraying and inductively coupled radio frequency plasma spraying techniques. Spherical powders with different particle size distributions were obtained after spheroidization. The effects of spherical glass powders on the mechanical properties of glass ionomer cements (GICs) were investigated. Results showed that the particle size distribution of the glass powders had a significant influence on the mechanical properties of GICs. Powders with a bimodal particle size distribution ensured a high packing density of glass ionomer cements, giving relatively high mechanical properties of GICs. GICs prepared by flame-spheroidized powders showed low strength values due to the loss of fine particles during flame spraying, leading to a low packing density and few metal ions reacting with polyacrylic acid to form cross-linking. GICs prepared by the nano-sized powders showed low strength because of the low bulk density of the nano-sized powders and hence low powder/liquid ratio of GICs.

  1. Monoenergetic electron parameters in a spheroid bubble model

    Science.gov (United States)

    Sattarian, H.; Sh., Rahmatallahpur; Tohidi, T.

    2013-02-01

    A reliable analytical expression for the potential of plasma waves with phase velocities near the speed of light is derived. The presented spheroid cavity model is more consistent than the previous spherical and ellipsoidal models and it explains the mono-energetic electron trajectory more accurately, especially at the relativistic region. The maximum energy of electrons is calculated and it is shown that the maximum energy of the spheroid model is less than that of the spherical model. The electron energy spectrum is also calculated and it is found that the energy distribution ratio of electrons ΔE/E for the spheroid model under the conditions reported here is half that of the spherical model and it is in good agreement with the experimental value in the same conditions. As a result, the quasi-mono-energetic electron output beam interacting with the laser plasma can be more appropriately described with this model.

  2. Merging and Splitting of Plasma Spheroids in a Dusty Plasma

    Science.gov (United States)

    Mikikian, Maxime; Tawidian, Hagop; Lecas, Thomas

    2012-12-01

    Dust particle growth in a plasma is a strongly disturbing phenomenon for the plasma equilibrium. It can induce many different types of low-frequency instabilities that can be experimentally observed, especially using high-speed imaging. A spectacular case has been observed in a krypton plasma where a huge density of dust particles is grown by material sputtering. The instability consists of well-defined regions of enhanced optical emission that emerge from the electrode vicinity and propagate towards the discharge center. These plasma spheroids have complex motions resulting from their mutual interaction that can also lead to the merging of two plasma spheroids into a single one. The reverse situation is also observed with the splitting of a plasma spheroid into two parts. These results are presented for the first time and reveal new behaviors in dusty plasmas.

  3. Monoenergetic electron parameters in a spheroid bubble model

    International Nuclear Information System (INIS)

    Sattarian, H.; Rahmatallahpur, Sh.; Tohidi, T.

    2013-01-01

    A reliable analytical expression for the potential of plasma waves with phase velocities near the speed of light is derived. The presented spheroid cavity model is more consistent than the previous spherical and ellipsoidal models and it explains the mono-energetic electron trajectory more accurately, especially at the relativistic region. The maximum energy of electrons is calculated and it is shown that the maximum energy of the spheroid model is less than that of the spherical model. The electron energy spectrum is also calculated and it is found that the energy distribution ratio of electrons ΔE/E for the spheroid model under the conditions reported here is half that of the spherical model and it is in good agreement with the experimental value in the same conditions. As a result, the quasi-mono-energetic electron output beam interacting with the laser plasma can be more appropriately described with this model. (physics of gases, plasmas, and electric discharges)

  4. CONCENTRIC MACLAURIN SPHEROID MODELS OF ROTATING LIQUID PLANETS

    International Nuclear Information System (INIS)

    Hubbard, W. B.

    2013-01-01

    I present exact expressions for the interior gravitational potential V of a system of N concentric constant-density (Maclaurin) spheroids. I demonstrate an iteration procedure to find a self-consistent solution for the shapes of the interfaces between spheroids, and for the interior gravitational potential. The external free-space potential, expressed as a multipole expansion, emerges as part of the self-consistent solution. The procedure is both simpler and more precise than perturbation methods. One can choose the distribution and mass densities of the concentric spheroids so as to reproduce a prescribed barotrope to a specified accuracy. I demonstrate the method's efficacy by comparing its results with several published test cases.

  5. Off-center point explosion in a spheroid

    International Nuclear Information System (INIS)

    Morita, Kazuhiko; Sakashita, Shiro

    1978-01-01

    An off-center point explosion in a spheroid with exponential or Gaussian density distribution is investigated by applying the generalized Laumbach and Probstein method. For a typical example, we calculate the explosion in a spheroid with the eccentricity e = 0.7. If the separation distance between the center of the spheroid and the explosion point is larger than three times of the density scale height, the shock wave may almost propagate toward the direction of the minor axis of symmetry, within the polar angle of 30 0 . The shock envelope elongates toward the same direction and may form a polar jet and/or a tilted jet. But, in the case of an explosion in the equatorial plane (perpendicular to the minor axis of symmetry), two plasmas with the same form may be ejected into two different directions with the angle smaller than 180 0 . Explosion models of double radio sources and related objects are suggested. (author)

  6. The development and structure of thick-walled, multicellular, aerial spores in Diheterospora chlamydosporia (=Verticillium chlamydosporium).

    Science.gov (United States)

    Cambell, W P; Griffiths, D A

    1975-07-01

    The aerial, thick-walled spores in Diheterospara chlamydosporia arose as terminal swellings on erect hyphae. Repeated septation of the continuously swelling spore resulted in a multicellular structure. Immediately after the onset of septation secondary wall material was laid down between the two-layered primary wall and the plasmalemma. The presence of secondary wall material indicates that the multicellular spore is a dictyochlamydospore and not an aleuriospore. The relationship between chlamydospores and aleuriospores in other fungi is discussed.

  7. Evolution of multicellularity coincided with increased diversification of cyanobacteria and the Great Oxidation Event

    Science.gov (United States)

    Schirrmeister, Bettina E.; de Vos, Jurriaan M.; Antonelli, Alexandre; Bagheri, Homayoun C.

    2013-01-01

    Cyanobacteria are among the most diverse prokaryotic phyla, with morphotypes ranging from unicellular to multicellular filamentous forms, including those able to terminally (i.e., irreversibly) differentiate in form and function. It has been suggested that cyanobacteria raised oxygen levels in the atmosphere around 2.45–2.32 billion y ago during the Great Oxidation Event (GOE), hence dramatically changing life on the planet. However, little is known about the temporal evolution of cyanobacterial lineages, and possible interplay between the origin of multicellularity, diversification of cyanobacteria, and the rise of atmospheric oxygen. We estimated divergence times of extant cyanobacterial lineages under Bayesian relaxed clocks for a dataset of 16S rRNA sequences representing the entire known diversity of this phylum. We tested whether the evolution of multicellularity overlaps with the GOE, and whether multicellularity is associated with significant shifts in diversification rates in cyanobacteria. Our results indicate an origin of cyanobacteria before the rise of atmospheric oxygen. The evolution of multicellular forms coincides with the onset of the GOE and an increase in diversification rates. These results suggest that multicellularity could have played a key role in triggering cyanobacterial evolution around the GOE. PMID:23319632

  8. Effects of irradiation and cisplatin on human glioma spheroids: inhibition of cell proliferation and cell migration

    NARCIS (Netherlands)

    Fehlauer, Fabian; Muench, Martina; Rades, Dirk; Stalpers, Lukas J. A.; Leenstra, Sieger; van der Valk, Paul; Slotman, Ben; Smid, Ernst J.; Sminia, Peter

    2005-01-01

    Investigation of cell migration and proliferation of human glioma cell line spheroids (CLS) and evaluation of morphology, apoptosis, and immunohistochemical expression of MIB-1, p53, and p21 of organotypic muticellular spheroids (OMS) following cisplatin (CDDP) and irradiation (RT). Spheroids of the

  9. Symbiotic Cell Differentiation and Cooperative Growth in Multicellular Aggregates.

    Directory of Open Access Journals (Sweden)

    Jumpei F Yamagishi

    2016-10-01

    Full Text Available As cells grow and divide under a given environment, they become crowded and resources are limited, as seen in bacterial biofilms and multicellular aggregates. These cells often show strong interactions through exchanging chemicals, as evident in quorum sensing, to achieve mutualism and division of labor. Here, to achieve stable division of labor, three characteristics are required. First, isogenous cells differentiate into several types. Second, this aggregate of distinct cell types shows better growth than that of isolated cells without interaction and differentiation, by achieving division of labor. Third, this cell aggregate is robust with respect to the number distribution of differentiated cell types. Indeed, theoretical studies have thus far considered how such cooperation is achieved when the ability of cell differentiation is presumed. Here, we address how cells acquire the ability of cell differentiation and division of labor simultaneously, which is also connected with the robustness of a cell society. For this purpose, we developed a dynamical-systems model of cells consisting of chemical components with intracellular catalytic reaction dynamics. The reactions convert external nutrients into internal components for cellular growth, and the divided cells interact through chemical diffusion. We found that cells sharing an identical catalytic network spontaneously differentiate via induction from cell-cell interactions, and then achieve division of labor, enabling a higher growth rate than that in the unicellular case. This symbiotic differentiation emerged for a class of reaction networks under the condition of nutrient limitation and strong cell-cell interactions. Then, robustness in the cell type distribution was achieved, while instability of collective growth could emerge even among the cooperative cells when the internal reserves of products were dominant. The present mechanism is simple and general as a natural consequence of

  10. The organotypic multicellular spheroid is a relevant three-dimensional model to study adenovirus replication and penetration in human tumors in vitro

    NARCIS (Netherlands)

    Grill, Jacques; Lamfers, Martine L. M.; van Beusechem, Victor W.; Dirven, Clemens M.; Pherai, D. Shareen; Kater, Mathijs; van der Valk, Paul; Vogels, Ronald; Vandertop, W. Peter; Pinedo, Herbert M.; Curiel, David T.; Gerritsen, Winald R.

    2002-01-01

    The use of adenoviruses for gene transfer and as oncolytic agents is currently receiving widespread attention. As specific constraints to adenovirus distribution and spread cannot be studied in cell cultures, there is a need for an in vitro three-dimensional (3D) model mimicking the in vivo biology

  11. The Evolution of COP9 Signalosome in Unicellular and Multicellular Organisms.

    Science.gov (United States)

    Barth, Emanuel; Hübler, Ron; Baniahmad, Aria; Marz, Manja

    2016-05-02

    The COP9 signalosome (CSN) is a highly conserved protein complex, recently being crystallized for human. In mammals and plants the COP9 complex consists of nine subunits, CSN 1-8 and CSNAP. The CSN regulates the activity of culling ring E3 ubiquitin and plays central roles in pleiotropy, cell cycle, and defense of pathogens. Despite the interesting and essential functions, a thorough analysis of the CSN subunits in evolutionary comparative perspective is missing. Here we compared 61 eukaryotic genomes including plants, animals, and yeasts genomes and show that the most conserved subunits of eukaryotes among the nine subunits are CSN2 and CSN5. This may indicate a strong evolutionary selection for these two subunits. Despite the strong conservation of the protein sequence, the genomic structures of the intron/exon boundaries indicate no conservation at genomic level. This suggests that the gene structure is exposed to a much less selection compared with the protein sequence. We also show the conservation of important active domains, such as PCI (proteasome lid-CSN-initiation factor) and MPN (MPR1/PAD1 amino-terminal). We identified novel exons and alternative splicing variants for all CSN subunits. This indicates another level of complexity of the CSN. Notably, most COP9-subunits were identified in all multicellular and unicellular eukaryotic organisms analyzed, but not in prokaryotes or archaeas. Thus, genes encoding CSN subunits present in all analyzed eukaryotes indicate the invention of the signalosome at the root of eukaryotes. The identification of alternative splice variants indicates possible "mini-complexes" or COP9 complexes with independent subunits containing potentially novel and not yet identified functions. © The Author 2016. Published by Oxford University Press on behalf of the Society for Molecular Biology and Evolution.

  12. Sulphur, zinc and carbon in the Sculptor dwarf spheroidal galaxy

    NARCIS (Netherlands)

    Skúladóttir, Ása

    2016-01-01

    The Sculptor dwarf spheroidal galaxy is a Milky Way satellite with predominantly old stellar population, and therefore the ideal target to study early chemical evolution. The chemical abundances of photospheres of stars reveal the composition of their birth environment; studying stars of different

  13. The early days of the Sculptor dwarf spheroidal galaxy

    NARCIS (Netherlands)

    Jablonka, P.; North, P.; Mashonkina, L.; Hill, V.; Revaz, Y.; Shetrone, M.; Starkenburg, E.; Irwin, M.; Tolstoy, E.; Battaglia, G.; Venn, K.; Helmi, A.; Primas, F.; François, P.

    2015-01-01

    We present the high-resolution spectroscopic study of five -3.9 ≤ [Fe/H] ≤ -2.5 stars in the Local Group dwarf spheroidal, Sculptor, thereby doubling the number of stars with comparable observations in this metallicity range. We carry out a detailed analysis of the chemical abundances of α, iron

  14. Spheroidal Integral Equations for Geodetic Inversion of Geopotential Gradients

    Science.gov (United States)

    Novák, Pavel; Šprlák, Michal

    2018-03-01

    The static Earth's gravitational field has traditionally been described in geodesy and geophysics by the gravitational potential (geopotential for short), a scalar function of 3-D position. Although not directly observable, geopotential functionals such as its first- and second-order gradients are routinely measured by ground, airborne and/or satellite sensors. In geodesy, these observables are often used for recovery of the static geopotential at some simple reference surface approximating the actual Earth's surface. A generalized mathematical model is represented by a surface integral equation which originates in solving Dirichlet's boundary-value problem of the potential theory defined for the harmonic geopotential, spheroidal boundary and globally distributed gradient data. The mathematical model can be used for combining various geopotential gradients without necessity of their re-sampling or prior continuation in space. The model extends the apparatus of integral equations which results from solving boundary-value problems of the potential theory to all geopotential gradients observed by current ground, airborne and satellite sensors. Differences between spherical and spheroidal formulations of integral kernel functions of Green's kind are investigated. Estimated differences reach relative values at the level of 3% which demonstrates the significance of spheroidal approximation for flattened bodies such as the Earth. The observation model can be used for combined inversion of currently available geopotential gradients while exploring their spectral and stochastic characteristics. The model would be even more relevant to gravitational field modelling of other bodies in space with more pronounced spheroidal geometry than that of the Earth.

  15. The dwarf spheroidal galaxies around the milky way

    NARCIS (Netherlands)

    Tolstoy, E.; Battaglia, G.; Helmi, A.; Irwin, M. J.; Hill, V.; Vallenari, A; Tantalo, R; Portinari, L; Moretti, A

    2007-01-01

    We review the progress of ESO/WFI Imaging and VLT/FLAMES spectroscopy of large numbers of individual stars in nearby dwarf spheroidal galaxies by the Dwarf Abundances and Radial-velocities Team (DART). These observations have allowed us to show that neither the kinematics nor the abundance nor the

  16. Optical properties of spherical and oblate spheroidal gold shell colloids

    NARCIS (Netherlands)

    Penninkhof, J.J.; Moroz, A.; van Blaaderen, A.; Polman, A.

    2008-01-01

    The surface plasmon modes of spherical and oblate spheroidal core−shell colloids composed of a 312 nm diameter silica core and a 20 nm thick Au shell are investigated. Large arrays of uniaxially aligned core−shell colloids with size aspect ratios ranging from 1.0 to 1.7 are fabricated using a novel

  17. Numerical simulations of the metallicity distribution in dwarf spheroidal galaxies

    NARCIS (Netherlands)

    Ripamonti, E.; Tolstoy, E.; Helmi, A.; Battaglia, G.; Abel, T.

    2006-01-01

    Abstract: Recent observations show that the number of stars with very low metallicities in the dwarf spheroidal satellites of the Milky Way is low, despite the low average metallicities of stars in these systems. We undertake numerical simulations of star formation and metal enrichment of dwarf

  18. The dynamical and chemical evolution of dwarf spheroidal galaxies

    NARCIS (Netherlands)

    Revaz, Y.; Jablonka, P.; Sawala, T.; Hill, V.; Letarte, B.; Irwin, M.; Battaglia, G.; Helmi, A.; Shetrone, M. D.; Tolstoy, E.; Venn, K. A.

    We present a large sample of fully self-consistent hydrodynamical Nbody/Tree-SPH simulations of isolated dwarf spheroidal galaxies (dSphs). It has enabled us to identify the key physical parameters and mechanisms at the origin of the observed variety in the Local Group dSph properties. The initial

  19. Estimation of scattering from a moist rough surface with spheroidal ...

    Indian Academy of Sciences (India)

    Administrator

    less than 5⋅5% of the magnetic wavelength. We empha- size that the surface deviation is responsible for scattering at a given electromagnetic wavelength. 2. Theoretical consideration (basic theory). We consider a horizontally rough surface with slight per- centage of moisture (2–4⋅5%) with spheroidal dust parti- cles.

  20. Void nucleation in spheroidized steels during tensile deformation

    International Nuclear Information System (INIS)

    Fisher, J.R. Jr.

    1980-04-01

    An investigation was conducted to determine the effects of various mechanical and material parameters on void formation at cementite particles in axisymmetric tensile specimens of spheroidized plain carbon steels. Desired microstructures for each of three steel types were obtained. Observations of void morphology with respect to various microstructural features were made using optical and scanning electron microscopy

  1. Media additives to promote spheroid circularity and compactness in hanging drop platform.

    Science.gov (United States)

    Leung, Brendan M; Lesher-Perez, Sasha Cai; Matsuoka, Toshiki; Moraes, Christopher; Takayama, Shuichi

    2015-02-01

    Three-dimensional spheroid cultures have become increasingly popular as drug screening platforms, especially with the advent of different high throughput spheroid forming technologies. However, comparing drug efficacy across different cell types in spheroid culture can be difficult due to variations in spheroid morphologies and transport characteristics. Improving the reproducibility of compact, circular spheroids contributes to standardizing and increasing the fidelity of the desired gradient profiles in these drug screening three-dimensional tissue cultures. In this study we discuss the role that circularity and compaction has on spheroids, and demonstrate the impact methylcellulose (MethoCel) and collagen additives in the culture media can contribute to more compact and circular spheroid morphology. We demonstrate that improved spheroid formation is not a simple function of increased viscosity of the different macromolecule additives, suggesting that other macromolecular characteristics contribute to improved spheroid formation. Of the various macromolecular additives tested for hanging drop culture, MethoCel provided the most desirable spheroid formation. Additionally, the higher viscosity of MethoCel-containing media improved the ease of imaging of cellular spheroids within hanging drop cultures by reducing motion-induced image blur.

  2. Fluid absorption related to ion transport in human airway epithelial spheroids

    DEFF Research Database (Denmark)

    Pedersen, P S; Holstein-Rathlou, N H; Larsen, P L

    1999-01-01

    , and amiloride inhibited both values. Fluid transport rates were calculated from repeated measurements of spheroid diameters. The results showed that 1) non-CF and CF spheroids absorbed fluid at identical rates (4.4 microl x cm(-2) x h(-1)), 2) amiloride inhibited fluid absorption to a lower residual level...... in non-CF than in CF spheroids, 3) Cl(-)-channel inhibitors increased fluid absorption in amiloride-treated non-CF spheroids to a level equal to that of amiloride-treated CF spheroids, 4) hydrochlorothiazide reduced the amiloride-insensitive fluid absorption in both non-CF and CF spheroids, and 5......) osmotic water permeabilities were equal in non-CF and CF spheroids ( approximately 27 x 10(-7) cm x s(-1) x atm(-1))....

  3. Plasma spheroidization of iron powders in a non-transferred DC thermal plasma jet

    International Nuclear Information System (INIS)

    Kumar, S.; Selvarajan, V.

    2008-01-01

    In this paper, the results of plasma spheroidization of iron powders using a DC non-transferred plasma spray torch are presented. The morphology of the processed powders was characterized through scanning electron microscopy (SEM) and optical microscopy (OM). The percentages of spheroidized powders were calculated by the shape factors such as the Irregularity Parameter (IP) and Roundness (RN). A maximum of 83% of spheroidization can be achieved. The spheroidization results are compared with the theoretical estimation and they are found to be in good agreement. The phase composition of the spheroidized powder was analyzed by XRD. The effect of plasma jet temperature and plasma gas flow rate on spheroidization is discussed. At low plasma gas flow rates and at high plasma jet temperatures, the percentage of spheroidization is high

  4. Structurally diverse natural products that cause potassium leakage trigger multicellularity in Bacillus subtilis.

    Science.gov (United States)

    López, Daniel; Fischbach, Michael A; Chu, Frances; Losick, Richard; Kolter, Roberto

    2009-01-06

    We report a previously undescribed quorum-sensing mechanism for triggering multicellularity in Bacillus subtilis. B. subtilis forms communities of cells known as biofilms in response to an unknown signal. We discovered that biofilm formation is stimulated by a variety of small molecules produced by bacteria--including the B. subtilis nonribosomal peptide surfactin--that share the ability to induce potassium leakage. Natural products that do not cause potassium leakage failed to induce multicellularity. Small-molecule-induced multicellularity was prevented by the addition of potassium, but not sodium or lithium. Evidence is presented that potassium leakage stimulates the activity of a membrane protein kinase, KinC, which governs the expression of genes involved in biofilm formation. We propose that KinC responds to lowered intracellular potassium concentration and that this is a quorum-sensing mechanism that enables B. subtilis to respond to related and unrelated bacteria.

  5. Transfer, imaging, and analysis plate for facile handling of 384 hanging drop 3D tissue spheroids.

    Science.gov (United States)

    Cavnar, Stephen P; Salomonsson, Emma; Luker, Kathryn E; Luker, Gary D; Takayama, Shuichi

    2014-04-01

    Three-dimensional culture systems bridge the experimental gap between in vivo and in vitro physiology. However, nonstandardized formation and limited downstream adaptability of 3D cultures have hindered mainstream adoption of these systems for biological applications, especially for low- and moderate-throughput assays commonly used in biomedical research. Here we build on our recent development of a 384-well hanging drop plate for spheroid culture to design a complementary spheroid transfer and imaging (TRIM) plate. The low-aspect ratio wells of the TRIM plate facilitated high-fidelity, user-independent, contact-based collection of hanging drop spheroids. Using the TRIM plate, we demonstrated several downstream analyses, including bulk tissue collection for flow cytometry, high-resolution low working-distance immersion imaging, and timely reagent delivery for enzymatic studies. Low working-distance multiphoton imaging revealed a cell type-dependent, macroscopic spheroid structure. Unlike ovarian cancer spheroids, which formed loose, disk-shaped spheroids, human mammary fibroblasts formed tight, spherical, and nutrient-limited spheroids. Beyond the applications we describe here, we expect the hanging drop spheroid plate and complementary TRIM plate to facilitate analyses of spheroids across the spectrum of throughput, particularly for bulk collection of spheroids and high-content imaging.

  6. Relations between pH, oxygen partial pressure and growth in cultured cell spheroids.

    Science.gov (United States)

    Carlsson, J; Acker, H

    1988-11-15

    The pH gradients, oxygen partial-pressure gradients and growth curves were measured for 7 different types of spheroids. Growth curves were measured in liquid overlay culture and thereafter the spheroids were attached to cover glasses and transferred to a chamber for micro-electrode measurements. The spheroids were randomly divided for pH or pO2 measurements which then were made under conditions as identical as possible. The decreases in pO2 and pH, delta pO2 and delta pH were calculated as the difference between the values in the culture medium and the values 200 micron inside the spheroids. Each type of spheroid had a certain relation between delta pO2 and delta pH. The human colon carcinoma HT29, the mouse mammary carcinoma EMT6 and the hamster lung V79-379A spheroids had high values of the quotient delta pO2/delta pH. The human thyroid carcinoma HTh7 spheroids and the 3 types of human glioma spheroids had lower quotients. There was a tendency for fast-growing spheroids to have high quotients. Two extreme types of spheroids, HT29 (high quotient) and U-118 MG (low quotient) were analyzed for lactate production and oxygen consumption. The U-118 MG spheroids produced about 3 times more lactate and consumed about 3 times less oxygen than the HT29 spheroids. The differences in lactate production could not be explained by differences in the pyruvate Km values of lactate dehydrogenase. The results indicate that there are significant metabolic differences between the spheroid systems studied.

  7. Bio-electrosprayed multicellular zebrafish embryos are viable and develop normally

    International Nuclear Information System (INIS)

    Clarke, Jonathan D W; Jayasinghe, Suwan N

    2008-01-01

    Bio-electrosprays are rapidly emerging as a viable protocol for directly engineering living cells. This communication reports the bio-electrospraying of multicellular organisms, namely zebrafish embryos. The results demonstrate that the bio-electrospray protocol fails to induce any embryological perturbations. In addition to analysing overall embryo morphology, we use transgenic embryos that express green fluorescent protein in specific brain neurons to determine that neuronal numbers and organization are completely normal. These results demonstrate that the bio-electrospraying protocol does not interfere with the complex gene regulation and cell movements required for the development of a multicellular organism. (communication)

  8. Dark matter annihilations search in dwarf spheroidal galaxies with fermi

    International Nuclear Information System (INIS)

    Farnier, C.; Nuss, E.; Cohen-Tanugi, J.

    2011-01-01

    Launched in June 2008, the Fermi Gamma-ray Telescope includes a pair conversion detector designed for the 20 MeV to ∼300GeV gamma-ray sky study, the Large Area Telescope (LAT). Operating in all-sky survey mode, its excellent sensitivity and angular resolution will allow either to discover or constrain a signal coming through the annihilation of dark matter particles. Predicted by cold dark matter scenarios as the largest clumps, dwarf spheroidal galaxies are amongst the most attractive targets for indirect search of dark matter by gamma-ray experiments. We present here an overview of the Fermi LAT Dark Matter and New Physics Working Group efforts in the searches of gamma-ray fluxes coming from WIMP pair annihilations in dwarf spheroidal galaxies.

  9. Magnetohydrodynamic stability of spheromak plasma in spheroidal flux conserver

    International Nuclear Information System (INIS)

    Kaneko, Shobu; Kamitani, Atsushi.

    1985-11-01

    The MHD equilibrium configurations of spheromak plasmas in a spheroidal flux conserver are determined by use of a pressure distribution whose derivative dp/dψ vanishes on the magnetic axis, and by use of an optimized distribution. Here p is the pressure and ψ is the flux function. These equilibria are shown to be stable for symmetric modes. The stability for localized modes is investigated by the Mercier criterion. The values of the maximum beta ratio β max are evaluated for both pressure distributions and are shown to become about two times larger by optimization. If the condition, q axis max are found to be less than 30 %. The oblate spheroidal flux conserver is shown to be better than the toroidal conserver with a rectangular cross section from the standpoint of stability. (author)

  10. A 2-D nucleation-growth model of spheroidal graphite

    International Nuclear Information System (INIS)

    Lacaze, Jacques; Bourdie, Jacques; Castro-Román, Manuel Jesus

    2017-01-01

    Analysis of recent experimental investigations, in particular by transmission electron microscopy, suggests spheroidal graphite grows by 2-D nucleation of new graphite layers at the outer surface of the nodules. These layers spread over the surface along the prismatic direction of graphite which is the energetically preferred growth direction of graphite when the apparent growth direction of the nodules is along the basal direction of graphite. 2-D nucleation-growth models first developed for precipitation of pure substances are then adapted to graphite growth from the liquid in spheroidal graphite cast irons. Lateral extension of the new graphite layers is controlled by carbon diffusion in the liquid. This allows describing quantitatively previous experimental results giving strong support to this approach.

  11. Mass transfer inside oblate spheroidal solids: modelling and simulation

    Directory of Open Access Journals (Sweden)

    J. E. F. Carmo

    2008-03-01

    Full Text Available A numerical solution of the unsteady diffusion equation describing mass transfer inside oblate spheroids, considering a constant diffusion coefficient and the convective boundary condition, is presented. The diffusion equation written in the oblate spheroidal coordinate system was used for a two-dimensional case. The finite-volume method was employed to discretize the basic equation. The linear equation set was solved iteratively using the Gauss-Seidel method. As applications, the effects of the Fourier number, the Biot number and the aspect ratio of the body on the drying rate and moisture content during the process are presented. To validate the methodology, results obtained in this work are compared with analytical results of the moisture content encountered in the literature and good agreement was obtained. The results show that the model is consistent and it may be used to solve cases such as those that include disks and spheres and/or those with variable properties with small modifications.

  12. Spheroidization of inorganic compounds by the LPPS method

    Czech Academy of Sciences Publication Activity Database

    Mastný, L.; Brožek, Vlastimil; Medřický, Jan; Marek, I.

    2017-01-01

    Roč. 13, č. 1 (2017), s. 162 ISSN 1336-7242. [Zjazd chemikov /69./. 11.09.2017-15.09.2017, Vysoké Tatry, Starý Smokovec] R&D Projects: GA ČR GA15-12145S Institutional support: RVO:61389021 Keywords : Water stabilized plasma * Liquid precursor plasma spraying * nanoparticles * spheroidization Subject RIV: BL - Plasma and Gas Discharge Physics OBOR OECD: Fluids and plasma physics (including surface physics)

  13. Sulphur, zinc and carbon in the Sculptor dwarf spheroidal galaxy

    OpenAIRE

    Skúladóttir, Ása

    2016-01-01

    The Sculptor dwarf spheroidal galaxy is a Milky Way satellite with predominantly old stellar population, and therefore the ideal target to study early chemical evolution. The chemical abundances of photospheres of stars reveal the composition of their birth environment; studying stars of different ages, therefore, provides insight into the chemical enrichment history of the galaxy in which they dwell. High-resolution spectra of 100 stars were used to further explore the chemical enrichment hi...

  14. Method for Processing Liver Spheroids Using an Automatic Tissue Processor

    Science.gov (United States)

    2016-05-01

    alcohol dehydration and hot liquid wax infiltration. After the water in the tissue is replaced with wax and cooled, it then becomes possible to cut...effective for processing and preparing microscopy slides of liver spheroids. The general process involved formalin fixation, dehydration in a...DPBS);  formalin (37% neutral buffer formaldehyde);  series of alcohol solutions: 70, 80, 95, and 100% ethanol in water; 2  xylene

  15. A study on the spheroidization by palsma spraying

    International Nuclear Information System (INIS)

    Jung, In Ha; Ji, C. G.; Bae, S. O.; Yoon, J. H.; Kwon, H. I.

    2001-01-01

    Spheroidization of a powder in thermal plasma is a plausible method for powder morphology treatment. In this experiment, the spheroidization fraction was decreased with increasing the particle size. The higher fraction of H2 gas and higher probe position resulted in a higher spheroidization fraction. From this study, it follows that the good result could be attained from smaller size particle with higher hydrogen gas fraction and higher probe position. Through the statistical assessment, the results concluded that the effect of hydrogen gas content has more influence than particle size, and the probe position affects lower than the others. The interaction between the probe position and hydrogen gas fraction affects more than the other interactions. This alluded that the result largely depended on the particle size and hydrogen gas fraction, and there was an interaction between probe position and hydrogen gas fraction. Therefore, when altering the H2 gas fraction is considered, altering the probe position should also be considered. The fractions of each effect were roughly 25% by particle size, 9% by probe position and 35% by H2 gas fraction. At the results of X-ray diffraction patterns of the spheroidized particles, the large particle exhibited higher content of tetragonal phase, while smaller particle showed nearly cubic phase. This seemed to be the quenching rate; larger particle might have more time to phase separation upon cooling that smaller one. X-ray analysis results confirmed a relation between the particle size and tetragonal phase content, when the particles were sprayed at the same conditions. When particles were the same size but sprayed at different conditions, different content of tetragonal phase exhibited according to the experienced temperatures of particles. A particle wasting longer time for cooling to room temperature showed much tetragonal content which phase is stable at low temperatures

  16. A study on the spheroidization by palsma spraying

    Energy Technology Data Exchange (ETDEWEB)

    Jung, In Ha; Ji, C. G.; Bae, S. O.; Yoon, J. H.; Kwon, H. I

    2001-01-01

    Spheroidization of a powder in thermal plasma is a plausible method for powder morphology treatment. In this experiment, the spheroidization fraction was decreased with increasing the particle size. The higher fraction of H2 gas and higher probe position resulted in a higher spheroidization fraction. From this study, it follows that the good result could be attained from smaller size particle with higher hydrogen gas fraction and higher probe position. Through the statistical assessment, the results concluded that the effect of hydrogen gas content has more influence than particle size, and the probe position affects lower than the others. The interaction between the probe position and hydrogen gas fraction affects more than the other interactions. This alluded that the result largely depended on the particle size and hydrogen gas fraction, and there was an interaction between probe position and hydrogen gas fraction. Therefore, when altering the H2 gas fraction is considered, altering the probe position should also be considered. The fractions of each effect were roughly 25% by particle size, 9% by probe position and 35% by H2 gas fraction. At the results of X-ray diffraction patterns of the spheroidized particles, the large particle exhibited higher content of tetragonal phase, while smaller particle showed nearly cubic phase. This seemed to be the quenching rate; larger particle might have more time to phase separation upon cooling that smaller one. X-ray analysis results confirmed a relation between the particle size and tetragonal phase content, when the particles were sprayed at the same conditions. When particles were the same size but sprayed at different conditions, different content of tetragonal phase exhibited according to the experienced temperatures of particles. A particle wasting longer time for cooling to room temperature showed much tetragonal content which phase is stable at low temperatures.

  17. Scaffold-free Prevascularized Microtissue Spheroids for Pulp Regeneration.

    Science.gov (United States)

    Dissanayaka, W L; Zhu, L; Hargreaves, K M; Jin, L; Zhang, C

    2014-12-01

    Creating an optimal microenvironment that mimics the extracellular matrix (ECM) of natural pulp and securing an adequate blood supply for the survival of cell transplants are major hurdles that need to be overcome in dental pulp regeneration. However, many currently available scaffolds fail to mimic essential functions of natural ECM. The present study investigated a novel approach involving the use of scaffold-free microtissue spheroids of dental pulp stem cells (DPSCs) prevascularized by human umbilical vein endothelial cells (HUVECs) in pulp regeneration. In vitro-fabricated microtissue spheroids were inserted into the canal space of tooth-root slices and were implanted subcutaneously into immunodeficient mice. Histological examination revealed that, after four-week implantation, tooth-root slices containing microtissue spheroids resulted in well-vascularized and cellular pulp-like tissues, compared with empty tooth-root slices, which were filled with only subcutaneous fat tissue. Immunohistochemical staining indicated that the tissue found in the tooth-root slices was of human origin, as characterized by the expression of human mitochondria, and contained odontoblast-like cells organized along the dentin, as assessed by immunostaining for nestin and dentin sialoprotein (DSP). Vascular structures formed by HUVECs in vitro were successfully anastomosed with the host vasculature upon transplantation in vivo, as shown by immunostaining for human CD31. Collectively, these findings demonstrate that prevascularized, scaffold-free, microtissue spheroids can successfully regenerate vascular dental pulp-like tissue and also highlight the significance of the microtissue microenvironment as an optimal environment for successful pulp-regeneration strategies. © International & American Associations for Dental Research.

  18. On the swimming motion of spheroidal magnetotactic bacteria

    Energy Technology Data Exchange (ETDEWEB)

    Cui Zhen; Kong Dali; Zhang Keke [Department of Mathematical Sciences, University of Exeter, Exeter EX4 4QF (United Kingdom); Pan Yongxin, E-mail: kzhang@ex.ac.uk [Institute of Geology and Geophysics, Chinese Academy of Sciences, Beijing (China)

    2012-10-15

    We investigate, via both theoretical and experimental methods, the swimming motion of magnetotactic bacteria having the shape of an elongated prolate spheroid in a viscous liquid under the influence of an imposed magnetic field. A fully three-dimensional Stokes flow, driven by the translation and rotation of a swimming bacterium, exerts a complicated viscous drag/torque on the motion of a non-spherical bacterium. By assuming that the body of the bacterium is non-deformable and that the interaction between different bacteria is weak and hence negligible, we have derived a system of 12 coupled nonlinear ordinary differential equations that govern both the motion and the orientation of a swimming spheroidal magnetotactic bacterium. The focus of the study is on how the shape of a non-spherical magnetotactic bacterium, marked by the size of its eccentricity, affects the pattern of its swimming motion. It is revealed that the pattern/speed of a swimming spheroidal magnetotactic bacterium is highly sensitive not only to the direction of its magnetic moment but also to its shape. We also compare the theoretical pattern obtained from the solutions of the 12 coupled differential equations with that observed in the laboratory experiments using the magnetotactic bacteria found in Lake Miyun near Beijing, China, showing that the observed pattern can be largely reproduced with an appropriate set of parameters in our theoretical model. (paper)

  19. Reparative Spheroids in HPV-Associated Chronic Cervicitis

    Directory of Open Access Journals (Sweden)

    Gennadiy T. Sukhikh

    2013-09-01

    Full Text Available Background: Spheroid cell structures (SCS described in cell culture are used to study cell-cell and cell-matrix interactions. However, the role of the SCS in the repair process in vivo remains unexplored. The aim of the study was to examine the cellular composition of the spherical structures and their functional significance in the repair of the squamous epithelium in human papilloma virus-associated chronic cervicitis (HPV-CC. Methods and Results: The cytology and biopsy materials from 223 patients with HPV-CC were subjected to molecular testing for HPV DNA by Real-Time Polymerase Chain Reaction (Real-Time PCR with genotyping and chromogenic in situ hybridization (CISH, as well as immunocytological and immunohistochemical analyses of p16INK4A, Ki67, SMA, Vimentin, CD34, E-cadherin, Oct4, CD44, CKW markers. In the stem cell niche zone, these spheroid structures were discovered having proliferative activity and showing signs of producing stem cells involved in the repair of the cervical mucosa in HPV-CC. Conclusion: The persistence of the HPV in the stem cell niche zone cells in the cervix determines the chronization of inflammation in this area, with the ability to perform pathological repair. The immunophenotype of the spheroid cell structures in the HPV-CC includes cells with signs of stem cells (‘stemness’ and the mesenchymal-epithelial transition.

  20. On the swimming motion of spheroidal magnetotactic bacteria

    International Nuclear Information System (INIS)

    Cui Zhen; Kong Dali; Zhang Keke; Pan Yongxin

    2012-01-01

    We investigate, via both theoretical and experimental methods, the swimming motion of magnetotactic bacteria having the shape of an elongated prolate spheroid in a viscous liquid under the influence of an imposed magnetic field. A fully three-dimensional Stokes flow, driven by the translation and rotation of a swimming bacterium, exerts a complicated viscous drag/torque on the motion of a non-spherical bacterium. By assuming that the body of the bacterium is non-deformable and that the interaction between different bacteria is weak and hence negligible, we have derived a system of 12 coupled nonlinear ordinary differential equations that govern both the motion and the orientation of a swimming spheroidal magnetotactic bacterium. The focus of the study is on how the shape of a non-spherical magnetotactic bacterium, marked by the size of its eccentricity, affects the pattern of its swimming motion. It is revealed that the pattern/speed of a swimming spheroidal magnetotactic bacterium is highly sensitive not only to the direction of its magnetic moment but also to its shape. We also compare the theoretical pattern obtained from the solutions of the 12 coupled differential equations with that observed in the laboratory experiments using the magnetotactic bacteria found in Lake Miyun near Beijing, China, showing that the observed pattern can be largely reproduced with an appropriate set of parameters in our theoretical model. (paper)

  1. Formation mechanism of spheroidal carbide in ultra-low carbon ductile cast iron

    Directory of Open Access Journals (Sweden)

    Bin-guo Fu

    2016-09-01

    Full Text Available The formation mechanism of the spheroidal carbide in the ultra-low carbon ductile cast iron fabricated by the metal mold casting technique was systematically investigated. The results demonstrated that the spheroidal carbide belonged to eutectic carbide and crystallized in the isolated eutectic liquid phase area. The formation process of the spheroidal carbide was related to the contact and the intersection between the primary dendrite and the secondary dendrite of austenite. The oxides of magnesium, rare earths and other elements can act as heterogeneous nucleation sites for the spheroidal carbide. It was also found that the amount of the spheroidal carbide would increase with an increase in carbon content. The cooling rate has an important influence on the spheroidal carbide under the same chemical composition condition.

  2. Porous Nb-Ti based alloy produced from plasma spheroidized powder

    OpenAIRE

    Li, Qijun; Zhang, Lin; Wei, Dongbin; Ren, Shubin; Qu, Xuanhui

    2017-01-01

    Spherical Nb-Ti based alloy powder was prepared by the combination of plasma spheroidization and mechanical alloying. Phase constituents, microstructure and surface state of the powder, and pore characteristics of the resulting porous alloy were investigated. The results show that the undissolved W and V in the mechanically alloyed powder is fully alloyed after spheroidization, and single β phase is achieved. Particle size of the spheroidized powder is in the range of 20–110 μm. With the decr...

  3. The Emergence of Multi-Cellular Robot Organisms through On-line On-board Evolution

    NARCIS (Netherlands)

    Weel, B.P.M.; Haasdijk, E.W.; Eiben, A.E.

    2012-01-01

    We investigate whether a swarm of robots can evolve controllers that cause aggregation into 'multi-cellular' robot organisms without a specific reward to do so. To this end, we create a world where aggregated robots receive more energy than individual ones and enable robots to evolve their

  4. Processing and characterization of multi-cellular monolithic bioceramics for bone regenerative scaffolds

    Science.gov (United States)

    Ari-Wahjoedi, Bambang; Ginta, Turnad Lenggo; Parman, Setyamartana; Abustaman, Mohd Zikri Ahmad

    2014-10-01

    Multicellular monolithic ceramic body is a ceramic material which has many gas or liquid passages partitioned by thin walls throughout the bulk material. There are many currently known advanced industrial applications of multicellular ceramics structures i.e. as supports for various catalysts, electrode support structure for solid oxide fuel cells, refractories, electric/electronic materials, aerospace vehicle re-entry heat shields and biomaterials for dental as well as orthopaedic implants by naming only a few. Multicellular ceramic bodies are usually made of ceramic phases such as mullite, cordierite, aluminum titanate or pure oxides such as silica, zirconia and alumina. What make alumina ceramics is excellent for the above functions are the intrinsic properties of alumina which are hard, wear resistant, excellent dielectric properties, resists strong acid and alkali attacks at elevated temperatures, good thermal conductivities, high strength and stiffness as well as biocompatible. In this work the processing technology leading to truly multicellular monolithic alumina ceramic bodies and their characterization are reported. Ceramic slip with 66 wt.% solid loading was found to be optimum as impregnant to the polyurethane foam template. Mullitic ceramic composite of alumina-sodium alumino disilicate-Leucite-like phases with bulk and true densities of 0.852 and 1.241 g cm-3 respectively, pore linear density of ±35 cm-1, linear and bulk volume shrinkages of 7-16% and 32 vol.% were obtained. The compressive strength and elastic modulus of the bioceramics are ≈0.5-1.0 and ≈20 MPa respectively.

  5. A Duo of Potassium-Responsive Histidine Kinases Govern the Multicellular Destiny of Bacillus subtilis

    NARCIS (Netherlands)

    Grau, Roberto R; de Oña, Paula; Kunert, Maritta; Leñini, Cecilia; Gallegos-Monterrosa, Ramses; Mhatre, Eisha; Vileta, Darío; Donato, Verónica; Hölscher, Theresa; Boland, Wilhelm; Kuipers, Oscar P; Kovács, Ákos T

    2015-01-01

    Multicellular biofilm formation and surface motility are bacterial behaviors considered mutually exclusive. However, the basic decision to move over or stay attached to a surface is poorly understood. Here, we discover that in Bacillus subtilis, the key root biofilm-controlling transcription factor

  6. Cellular Particle Dynamics simulation of biomechanical relaxation processes of multi-cellular systems

    Science.gov (United States)

    McCune, Matthew; Kosztin, Ioan

    2013-03-01

    Cellular Particle Dynamics (CPD) is a theoretical-computational-experimental framework for describing and predicting the time evolution of biomechanical relaxation processes of multi-cellular systems, such as fusion, sorting and compression. In CPD, cells are modeled as an ensemble of cellular particles (CPs) that interact via short range contact interactions, characterized by an attractive (adhesive interaction) and a repulsive (excluded volume interaction) component. The time evolution of the spatial conformation of the multicellular system is determined by following the trajectories of all CPs through numerical integration of their equations of motion. Here we present CPD simulation results for the fusion of both spherical and cylindrical multi-cellular aggregates. First, we calibrate the relevant CPD model parameters for a given cell type by comparing the CPD simulation results for the fusion of two spherical aggregates to the corresponding experimental results. Next, CPD simulations are used to predict the time evolution of the fusion of cylindrical aggregates. The latter is relevant for the formation of tubular multi-cellular structures (i.e., primitive blood vessels) created by the novel bioprinting technology. Work supported by NSF [PHY-0957914]. Computer time provided by the University of Missouri Bioinformatics Consortium.

  7. X-ray microscopy of living multicellular organisms with the Prague Asterix Iodine Laser System

    Czech Academy of Sciences Publication Activity Database

    Desai, T.; Batani, D.; Bernadinello, A.; Poletti, G.; Orsini, F.; Ullschmied, Jiří; Juha, Libor; Skála, Jiří; Králiková, Božena; Krouský, Eduard; Pfeifer, Miroslav; Kadlec, Christelle; Mocek, Tomáš; Präg R., Ansgar; Renner, Oldřich; Cotelli, F.; Lamia, C. L.; Zullini, A.

    2003-01-01

    Roč. 21, č. 4 (2003), s. 511-516 ISSN 0263-0346 R&D Projects: GA MŠk LN00A100 Institutional research plan: CEZ:AV0Z2043910 Keywords : atomic force miscroscopy, laser-produced plasmas, multicellular microorganisms Subject RIV: BH - Optics, Masers, Lasers Impact factor: 0.646, year: 2003

  8. Targeting tumor multicellular aggregation through IGPR-1 inhibits colon cancer growth and improves chemotherapy.

    Science.gov (United States)

    Woolf, N; Pearson, B E; Bondzie, P A; Meyer, R D; Lavaei, M; Belkina, A C; Chitalia, V; Rahimi, N

    2017-09-18

    Adhesion to extracellular matrix (ECM) is crucially important for survival of normal epithelial cells as detachment from ECM triggers specific apoptosis known as anoikis. As tumor cells lose the requirement for anchorage to ECM, they rely on cell-cell adhesion 'multicellular aggregation' for survival. Multicellular aggregation of tumor cells also significantly determines the sensitivity of tumor cells to the cytotoxic effects of chemotherapeutics. In this report, we demonstrate that expression of immunoglobulin containing and proline-rich receptor-1 (IGPR-1) is upregulated in human primary colon cancer. Our study demonstrates that IGPR-1 promotes tumor multicellular aggregation, and interfering with its adhesive function inhibits multicellular aggregation and, increases cell death. IGPR-1 supports colon carcinoma tumor xenograft growth in mouse, and inhibiting its activity by shRNA or blocking antibody inhibits tumor growth. More importantly, IGPR-1 regulates sensitivity of tumor cells to the chemotherapeutic agent, doxorubicin/adriamycin by a mechanism that involves doxorubicin-induced AKT activation and phosphorylation of IGPR-1 at Ser220. Our findings offer novel insight into IGPR-1's role in colorectal tumor growth, tumor chemosensitivity, and as a possible novel anti-cancer target.

  9. Resistance of multicellular aggregates to pharmorubicin observed in human hepatocarcinoma cells

    Directory of Open Access Journals (Sweden)

    Z. Jianmin

    2002-02-01

    Full Text Available The objective of the present study was to investigate the multicellular resistance of human hepatocarcinoma cells BEL-7402 to pharmorubicin. Cells (1 x 10(4 and 200 microcarrier Cytodex-3 beads were seeded onto a 24-well plate and cultured in RPMI 1640 medium. After the formation of multicellular aggregates, morphology and cell viability were analyzed by scanning electron microscopy, transmission electron microscopy and flow cytometry, respectively. The IC50 was determined by flow cytometry and MTT assay after the cells cultured in aggregates and monolayers were treated with pharmorubicin. The culture products exhibited structural characteristics somewhat similar to those of trabecular hepatocarcinoma in vivo. Among the microcarriers, cells were organized into several layers. Intercellular spaces were 0.5-2.0 µm wide and filled with many microvilli. The percent of viable cells was 87%. The cells cultured as multicellular aggregates were resistant to pharmorubicin with IC50 4.5-fold and 7.7-fold that of monolayer culture as determined by flow cytometry and MTT assay, respectively. This three-dimensional culture model may be used to investigate the mechanisms of multicellular drug resistance of hepatocarcinoma and to screen new anticancer drugs.

  10. Cell Spheroids with Enhanced Aggressiveness to Mimic Human Liver Cancer In Vitro and In Vivo.

    Science.gov (United States)

    Jung, Hong-Ryul; Kang, Hyun Mi; Ryu, Jea-Woon; Kim, Dae-Soo; Noh, Kyung Hee; Kim, Eun-Su; Lee, Ho-Joon; Chung, Kyung-Sook; Cho, Hyun-Soo; Kim, Nam-Soon; Im, Dong-Soo; Lim, Jung Hwa; Jung, Cho-Rok

    2017-09-05

    We fabricated a spheroid-forming unit (SFU) for efficient and economic production of cell spheroids. We optimized the protocol for generating large and homogenous liver cancer cell spheroids using Huh7 hepatocellular carcinoma (HCC) cells. The large Huh7 spheroids showed apoptotic and proliferative signals in the centre and at the surface, respectively. In particular, hypoxia-induced factor-1 alpha (HIF-1α) and ERK signal activation were detected in the cell spheroids. To diminish core necrosis and increase the oncogenic character, we co-cultured spheroids with 2% human umbilical vein endothelial cells (HUVECs). HUVECs promoted proliferation and gene expression of HCC-related genes and cancer stem cell markers in the Huh7 spheroidsby activating cytokine signalling, mimicking gene expression in liver cancer. HUVECs induced angiogenesis and vessel maturation in Huh7 spheroids in vivo by activating epithelial-mesenchymal transition and angiogenic pathways. The large Huh7 cell spheroids containing HUVECs survived at higher concentrations of anti-cancer drugs (doxorubicin and sorafenib) than did monolayer cells. Our large cell spheroid provides a useful in vitro HCC model to enable intuitive observation for anti-cancer drug testing.

  11. Measuring the light scattering and orientation of a spheroidal particle using in-line holography.

    Science.gov (United States)

    Seo, Kyung Won; Byeon, Hyeok Jun; Lee, Sang Joon

    2014-07-01

    The light scattering properties of a horizontally and vertically oriented spheroidal particle under laser illumination are experimentally investigated using digital in-line holography. The reconstructed wave field shows the bright singular points as a result of the condensed beam formed by a transparent spheroidal particle acting as a lens. The in-plane (θ) and out-of-plane (ϕ) rotating angles of an arbitrarily oriented spheroidal particle are measured by using these scattering properties. As a feasibility test, the 3D orientation of a transparent spheroidal particle suspended in a microscale pipe flow is successfully reconstructed by adapting the proposed method.

  12. Exposing diversity

    DEFF Research Database (Denmark)

    Nørtoft, Kamilla; Nordentoft, Helle Merete

    professionals´ meetings with patients and relatives. In the paper we draw data from focus group discussions with interdisciplinary groups of health care professionals working in the area of care for older people. The video narratives used to initiate discussions are developed through ethnographic fieldwork...... in the homes of older people and in pedagogical institutions targeting older people. In the paper we look at the potentials and challenges in working with ethnographic video narratives as a pedagogical tool. Our findings indicate that the use of video narratives has the potential to expose the diversity...... focus on their own professional discipline and its tasks 2) stimulates collaborative learning when they discuss their different interpretations of the ethnographic video narratives and achieve a deeper understanding of each other’s work and their clients’ lifeworlds, which might lead to a better...

  13. Optimal formation of genetically modified and functional pancreatic islet spheroids by using hanging-drop strategy.

    Science.gov (United States)

    Kim, H J; Alam, Z; Hwang, J W; Hwang, Y H; Kim, M J; Yoon, S; Byun, Y; Lee, D Y

    2013-03-01

    Rejection and hypoxia are important factors causing islet loss at an early stage after pancreatic islet transplantation. Recently, islets have been dissociated into single cells for reaggregation into so-called islet spheroids. Herein, we used a hanging-drop strategy to form islet spheroids to achieve functional equivalence to intact islets. To obtain single islet cells, we dissociated islets with trypsin-EDTA digestion for 10 minutes. To obtain spheroids, we dropped various numbers of single cells (125, 250, or 500 cells/30 μL drop) onto a Petri dish, that was inverted for incubation in humidified air containing 5% CO(2) at 37 °C for 7 days. The aggregated spheroids in the droplets were harvested for further culture. The size of the aggregated islet spheroids depended on the number of single cells (125-500 cells/30 μL droplet). Their morphology was similar to that of intact islets without any cellular damage. When treated with various concentrations of glucose to evaluate responsiveness, their glucose-mediated stimulation index value was similar to that of intact islets, an observation that was attributed to strong cell-to-cell interactions in islet spheroids. However, islet spheroids aggregated in general culture dishes showed abnormal glucose responsiveness owing to weak cell-to-cell interactions. Cell-to-cell interactions in islet spheroids were confirmed with an anti-connexin-36 monoclonal antibody. Finally, nonviral poly(ethylene imine)-mediated interleukin-10 cytokine gene delivered beforehand into dissociated single cells before formation of islet spheroids increased the gene transfection efficacy and interleukin-10 secretion from islet spheroids >4-fold compared with intact islets. These results demonstrated the potential application of genetically modified, functional islet spheroids with of controlled size and morphology using an hanging-drop technique. Copyright © 2013 Elsevier Inc. All rights reserved.

  14. In vitro characterization of self-assembled anterior cruciate ligament cell spheroids for ligament tissue engineering.

    Science.gov (United States)

    Hoyer, M; Meier, C; Breier, A; Hahner, J; Heinrich, G; Drechsel, N; Meyer, M; Rentsch, C; Garbe, L-A; Ertel, W; Lohan, A; Schulze-Tanzil, G

    2015-03-01

    Tissue engineering of an anterior cruciate ligament (ACL) implant with functional enthesis requires site-directed seeding of different cell types on the same scaffold. Therefore, we studied the suitability of self-assembled three-dimensional spheroids generated by lapine ACL ligament fibroblasts for directed scaffold colonization. The spheroids were characterized in vitro during 14 days in static and 7 days in dynamic culture. Size maintenance of self-assembled spheroids, the vitality, the morphology and the expression pattern of the cells were monitored. Additionally, we analyzed the total sulfated glycosaminoglycan, collagen contents and the expression of the ligament components type I collagen, decorin and tenascin C on protein and for COL1A1, DCN and TNMD on gene level in the spheroids. Subsequently, the cell colonization of polylactide-co-caprolactone [P(LA-CL)] and polydioxanone (PDS) polymer scaffolds was assessed in response to a directed, spheroid-based seeding technique. ACL cells were able to self-assemble spheroids and survive over 14 days. The spheroids decreased in size but not in cellularity depending on the culture time and maintained or even increased their differentiation state. The area of P[LA-CL] scaffolds, colonized after 14 days by the cells of one spheroid, was in average 4.57 ± 2.3 mm(2). Scaffolds consisting of the polymer P[LA-CL] were more suitable for colonization by spheroids than PDS embroideries. We conclude that ACL cell spheroids are suitable as site-directed seeding strategy for scaffolds in ACL tissue engineering approaches and recommend the use of freshly assembled spheroids for scaffold colonization, due to their balanced proliferation and differentiation.

  15. Generating Chondromimetic Mesenchymal Stem Cell Spheroids by Regulating Media Composition and Surface Coating.

    Science.gov (United States)

    Sridharan, BanuPriya; Laflin, Amy D; Detamore, Michael S

    2018-04-01

    Spheroids of mesenchymal stem cells (MSCs) in cartilage tissue engineering have been shown to enhance regenerative potential owing to their 3D structure. In this study, we explored the possibility of priming spheroids under different media to replace the use of inductive surface coatings for chondrogenic differentiation. Rat bone marrow-derived MSCs were organized into cell spheroids by the hanging drop technique and subsequently cultured on hyaluronic acid (HA) coated or non-coated well plates under different cell media conditions. Endpoint analysis included cell viability, DNA and Glycosaminoglycan (GAG) and collagen content, gene expression and immunohistochemistry. For chondrogenic applications, MSC spheroids derived on non-coated surfaces outperformed the spheroids derived from HA-coated surfaces in matrix synthesis and collagen II gene expression. Spheroids on non-coated surfaces gave rise to the highest collagen and GAG when primed with medium containing insulin-like growth factor (IGF) for 1 week during spheroid formation. Spheroids that were grown in chondroinductive raw material-inclusive media such as aggrecan or chondroitin sulfate exhibited the highest Collagen II gene expression in the non-coated surface at 1 week. Media priming by growth factors and raw materials might be a more predictive influencer of chondrogenesis compared to inductive-surfaces. Such tailored bioactivity of the stem cell spheroids in the stage of the spheroid formation may give rise to a platform technology that may eventually produce spheroids capable of chondrogenesis achieved by mere media manipulation, skipping the need for additional culture on a modified surface, that paves the way for cost-effective technologies.

  16. Chitosan derived co-spheroids of neural stem cells and mesenchymal stem cells for neural regeneration.

    Science.gov (United States)

    Han, Hao-Wei; Hsu, Shan-Hui

    2017-10-01

    Chitosan has been considered as candidate biomaterials for neural applications. The effective treatment of neurodegeneration or injury to the central nervous system (CNS) is still in lack nowadays. Adult neural stem cells (NSCs) represents a promising cell source to treat the CNS diseases but they are limited in number. Here, we developed the core-shell spheroids of NSCs (shell) and mesenchymal stem cells (MSCs, core) by co-culturing cells on the chitosan surface. The NSCs in chitosan derived co-spheroids displayed a higher survival rate than those in NSC homo-spheroids. The direct interaction of NSCs with MSCs in the co-spheroids increased the Notch activity and differentiation tendency of NSCs. Meanwhile, the differentiation potential of MSCs in chitosan derived co-spheroids was significantly enhanced toward neural lineages. Furthermore, NSC homo-spheroids and NSC/MSC co-spheroids derived on chitosan were evaluated for their in vivo efficacy by the embryonic and adult zebrafish brain injury models. The locomotion activity of zebrafish receiving chitosan derived NSC homo-spheroids or NSC/MSC co-spheroids was partially rescued in both models. Meanwhile, the higher survival rate was observed in the group of adult zebrafish implanted with chitosan derived NSC/MSC co-spheroids as compared to NSC homo-spheroids. These evidences indicate that chitosan may provide an extracellular matrix-like environment to drive the interaction and the morphological assembly between NSCs and MSCs and promote their neural differentiation capacities, which can be used for neural regeneration. Copyright © 2017 Elsevier B.V. All rights reserved.

  17. Three-dimensional tumor spheroids for in vitro analysis of bacteria as gene delivery vectors in tumor therapy.

    Science.gov (United States)

    Osswald, Annika; Sun, Zhongke; Grimm, Verena; Ampem, Grace; Riegel, Karin; Westendorf, Astrid M; Sommergruber, Wolfgang; Otte, Kerstin; Dürre, Peter; Riedel, Christian U

    2015-12-12

    Several studies in animal models demonstrated that obligate and facultative anaerobic bacteria of the genera Bifidobacterium, Salmonella, or Clostridium specifically colonize solid tumors. Consequently, these and other bacteria are discussed as live vectors to deliver therapeutic genes to inhibit tumor growth. Therapeutic approaches for cancer treatment using anaerobic bacteria have been investigated in different mouse models. In the present study, solid three-dimensional (3D) multicellular tumor spheroids (MCTS) of the colorectal adenocarcinoma cell line HT-29 were generated and tested for their potential to study prodrug-converting enzyme therapies using bacterial vectors in vitro. HT-29 MCTS resembled solid tumors displaying all relevant features with an outer zone of proliferating cells and hypoxic and apoptotic regions in the core. Upon incubation with HT-29 MCTS, Bifidobacterium bifidum S17 and Salmonella typhimurium YB1 selectively localized, survived and replicated in hypoxic areas inside MCTS. Furthermore, spores of the obligate anaerobe Clostridium sporogenes germinated in these hypoxic areas. To further evaluate the potential of MCTS to investigate therapeutic approaches using bacteria as gene delivery vectors, recombinant bifidobacteria expressing prodrug-converting enzymes were used. Expression of a secreted cytosine deaminase in combination with 5-fluorocytosine had no effect on growth of MCTS due to an intrinsic resistance of HT-29 cells to 5-fluorouracil, i.e. the converted drug. However, a combination of the prodrug CB1954 and a strain expressing a secreted chromate reductase effectively inhibited MCTS growth. Collectively, the presented results indicate that MCTS are a suitable and reliable model to investigate live bacteria as gene delivery vectors for cancer therapy in vitro.

  18. Supermassive Black Holes and Their Host Spheroids. I. Disassembling Galaxies

    Science.gov (United States)

    Savorgnan, G. A. D.; Graham, A. W.

    2016-01-01

    Several recent studies have performed galaxy decompositions to investigate correlations between the black hole mass and various properties of the host spheroid, but they have not converged on the same conclusions. This is because their models for the same galaxy were often significantly different and not consistent with each other in terms of fitted components. Using 3.6 μm Spitzer imagery, which is a superb tracer of the stellar mass (superior to the K band), we have performed state-of-the-art multicomponent decompositions for 66 galaxies with directly measured black hole masses. Our sample is the largest to date and, unlike previous studies, contains a large number (17) of spiral galaxies with low black hole masses. We paid careful attention to the image mosaicking, sky subtraction, and masking of contaminating sources. After a scrupulous inspection of the galaxy photometry (through isophotal analysis and unsharp masking) and—for the first time—2D kinematics, we were able to account for spheroids large-scale, intermediate-scale, and nuclear disks bars rings spiral arms halos extended or unresolved nuclear sources; and partially depleted cores. For each individual galaxy, we compared our best-fit model with previous studies, explained the discrepancies, and identified the optimal decomposition. Moreover, we have independently performed one-dimensional (1D) and two-dimensional (2D) decompositions and concluded that, at least when modeling large, nearby galaxies, 1D techniques have more advantages than 2D techniques. Finally, we developed a prescription to estimate the uncertainties on the 1D best-fit parameters for the 66 spheroids that takes into account systematic errors, unlike popular 2D codes that only consider statistical errors.

  19. SUPERMASSIVE BLACK HOLES AND THEIR HOST SPHEROIDS. I. DISASSEMBLING GALAXIES

    Energy Technology Data Exchange (ETDEWEB)

    Savorgnan, G. A. D.; Graham, A. W., E-mail: gsavorgn@astro.swin.edu.au [Centre for Astrophysics and Supercomputing, Swinburne University of Technology, Hawthorn, Victoria 3122 (Australia)

    2016-01-15

    Several recent studies have performed galaxy decompositions to investigate correlations between the black hole mass and various properties of the host spheroid, but they have not converged on the same conclusions. This is because their models for the same galaxy were often significantly different and not consistent with each other in terms of fitted components. Using 3.6 μm Spitzer imagery, which is a superb tracer of the stellar mass (superior to the K band), we have performed state-of-the-art multicomponent decompositions for 66 galaxies with directly measured black hole masses. Our sample is the largest to date and, unlike previous studies, contains a large number (17) of spiral galaxies with low black hole masses. We paid careful attention to the image mosaicking, sky subtraction, and masking of contaminating sources. After a scrupulous inspection of the galaxy photometry (through isophotal analysis and unsharp masking) and—for the first time—2D kinematics, we were able to account for spheroids; large-scale, intermediate-scale, and nuclear disks; bars; rings; spiral arms; halos; extended or unresolved nuclear sources; and partially depleted cores. For each individual galaxy, we compared our best-fit model with previous studies, explained the discrepancies, and identified the optimal decomposition. Moreover, we have independently performed one-dimensional (1D) and two-dimensional (2D) decompositions and concluded that, at least when modeling large, nearby galaxies, 1D techniques have more advantages than 2D techniques. Finally, we developed a prescription to estimate the uncertainties on the 1D best-fit parameters for the 66 spheroids that takes into account systematic errors, unlike popular 2D codes that only consider statistical errors.

  20. SUPERMASSIVE BLACK HOLES AND THEIR HOST SPHEROIDS. I. DISASSEMBLING GALAXIES

    International Nuclear Information System (INIS)

    Savorgnan, G. A. D.; Graham, A. W.

    2016-01-01

    Several recent studies have performed galaxy decompositions to investigate correlations between the black hole mass and various properties of the host spheroid, but they have not converged on the same conclusions. This is because their models for the same galaxy were often significantly different and not consistent with each other in terms of fitted components. Using 3.6 μm Spitzer imagery, which is a superb tracer of the stellar mass (superior to the K band), we have performed state-of-the-art multicomponent decompositions for 66 galaxies with directly measured black hole masses. Our sample is the largest to date and, unlike previous studies, contains a large number (17) of spiral galaxies with low black hole masses. We paid careful attention to the image mosaicking, sky subtraction, and masking of contaminating sources. After a scrupulous inspection of the galaxy photometry (through isophotal analysis and unsharp masking) and—for the first time—2D kinematics, we were able to account for spheroids; large-scale, intermediate-scale, and nuclear disks; bars; rings; spiral arms; halos; extended or unresolved nuclear sources; and partially depleted cores. For each individual galaxy, we compared our best-fit model with previous studies, explained the discrepancies, and identified the optimal decomposition. Moreover, we have independently performed one-dimensional (1D) and two-dimensional (2D) decompositions and concluded that, at least when modeling large, nearby galaxies, 1D techniques have more advantages than 2D techniques. Finally, we developed a prescription to estimate the uncertainties on the 1D best-fit parameters for the 66 spheroids that takes into account systematic errors, unlike popular 2D codes that only consider statistical errors

  1. Formation and field-driven dynamics of nematic spheroids.

    Science.gov (United States)

    Fu, Fred; Abukhdeir, Nasser Mohieddin

    2017-07-19

    Unlike the canonical application of liquid crystals (LCs), LC displays, emerging technologies based on LC materials are increasingly leveraging the presence of nanoscale defects. The inherent nanoscale characteristics of LC defects present both significant opportunities as well as barriers for the application of this fascinating class of materials. Simulation-based approaches to the study of the effects of confinement and interface anchoring conditions on LC domains has resulted in significant progress over the past decade, where simulations are now able to access experimentally-relevant length scales while simultaneously capturing nanoscale defect structures. In this work, continuum simulations were performed in order to study the dynamics of micron-scale nematic LC spheroids of varying shape. Nematic spheroids are one of the simplest inherently defect-containing LC structures and are relevant to polymer-dispersed LC-based "smart" window technology. Simulation results include nematic phase formation and external field-switching dynamics of nematic spheroids ranging in shape from oblate to prolate. Results include both qualitative and quantitative insight into the complex coupling of nanoscale defect dynamics and structure transitions to micron-scale reorientation. Dynamic mechanisms are presented and related to structural transitions in LC defects present in the nematic domain. Domain-averaged metrics including order parameters and response times are determined for a range of experimentally-accessible electric field strengths. These results have both fundamental and technological relevance, in that increased understanding of LC dynamics in the presence of defects is a key barrier to continued advancement in the field.

  2. Combination therapy with 1,3-bis(2-chloroethyl)-1-nitrosourea and low dose rate radiation in the 9L rat brain tumor and spheroid models: implications for brain tumor brachytherapy

    International Nuclear Information System (INIS)

    Gutin, P.H.; Bernstein, M.; Sano, Y.; Deen, D.F.

    1984-01-01

    The effects of combination treatment with 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU) and low dose rate radiation were studied in the 9L rat brain tumor in vivo model and the 9L multicellular tumor spheroid model. F-344 rats bearing intracerebral 9L gliosarcomas were implanted with removable 125 I sources. Minimal (peripheral) tumor doses of 6387 rad produced an increased life-span (ILS) of 28% over that of control rats implanted with dummy sources, BCNU alone (13.3 mg/kg) produced in an ILS of 67%, and combination treatment with BCNU and implanted 125 I sources produced an ILS of 167%. As measured by a colony-forming efficiency assay, the greatest cell kill in 9L spheroids occurred when BCNU was administered 24 hours before irradiation from a 137 Cs source at a low dose rate of 5 rad/minute. At a higher dose rate of 210 rad/minute, the time dependence of the effects of combination treatment was identical and therefore independent of dose rate

  3. WFPC2 Observations of the URSA Minor Dwarf Spheroidal Galaxy

    Science.gov (United States)

    Mighell, Kenneth J.; Burke, Christopher J.

    1999-01-01

    We present our analysis of archival Hubble Space Telescope Wide Field Planetary Camera 2 (WFPC2) observations in F555W (approximately V) and F814W (approximately I) of the central region of the Ursa Minor dwarf spheroidal galaxy. The V versus V - I color-magnitude diagram features a sparsely populated blue horizontal branch, a steep thin red giant branch, and a narrow subgiant branch. The main sequence reaches approximately 2 magnitudes below the main-sequence turnoff (V(sup UMi, sub TO) approximately equals 23.27 +/- 0.11 mag) of the median stellar population. We compare the fiducial sequence of the Galactic globular cluster M92 (NGC 6341). The excellent match between Ursa Minor and M92 confirms that the median stellar population of the UMi dSph galaxy is metal poor ([Fe/H](sub UMi) approximately equals [Fe/H](sub M92) approximately equals -2.2 dex) and ancient (age(sub UMi)approximately equalsage(sub M92) approximately equals 14 Gyr). The B - V reddening and the absorption in V are estimated to be E(B - V) = 0.03 +/- 0.01 mag and A(sup UMi, sub V) = 0.09 +/- 0.03 mag. A new estimate of the distance modulus of Ursa Minor, (m - M)(sup UMi, sub 0) = 19.18 +/- 0.12 mag, has been derived based on fiducial-sequence fitting M92 [DELTA.V(sub UMi - M92) = 4.60 +/- 0.03 mag and DELTA(V - I)(sub UMi - M92) = 0.010 +/- 0.005 mag] and the adoption of the apparent V distance modulus for M92 of (m - M)(sup M92, sub V) = 14.67 +/- 0.08 mag (Pont et al. 1998, A&A, 329, 87). The Ursa Minor dwarf spheroidal galaxy is then at a distance of 69 +/- 4 kpc from the Sun. These HST observations indicate that Ursa Minor has had a very simple star formation history consisting mainly of a single major burst of star formation about 14 Gyr ago which lasted approximately stars in the central region Ursa Minor dwarf spheroidal galaxy are ancient. If the ancient Galactic globular clusters, like M92, formed concurrently with the early formation of the Milky Way galaxy itself, then the Ursa Minor

  4. PREDICTION OF THE EXTREMAL SHAPE FACTOR OF SPHEROIDAL PARTICLES

    Directory of Open Access Journals (Sweden)

    Daniel Hlubinka

    2011-05-01

    Full Text Available In the stereological unfolding problem for spheroidal particles the extremal shape factor is predicted. The theory of extreme values has been used to show that extremes of the planar shape factor of particle sections tend to the same limit distribution as extremes of the original shape factor for both the conditional and marginal distribution. Attention is then paid to the extreme shape factor conditioned by the particle size. Normalizing constants are evaluated for a parametric model and the numerical procedure is tested on real data from metallography.

  5. Modes of spheroidal ion plasmas at the Brillouin limit

    International Nuclear Information System (INIS)

    Tinkle, M. D.; Greaves, R. G.; Surko, C. M.

    1995-01-01

    Brillouin-density pure ion plasmas have been generated in a quadrupole Penning tray by electron-beam ionization of a low-pressure gas. Large, spheroidal, steady-state plasmas are produced that extend to one of the trap electrodes. With the density fixed at the Brillouin limit by the high ion production rate, the electrode potentials determine the plasma shape. The frequencies of azimuthally propagating cyclotron and diocotron modes are found to vary significantly with the plasma aspect ratio. For oblate plasmas, we are able to test theoretical predictions of a simple fluid model, and the frequencies are in good agreement

  6. Differential Superiority of Heavy Charged-Particle Irradiation to X-Rays: Studies on Biological Effectiveness and Side Effect Mechanisms in Multicellular Tumor and Normal Tissue Models

    Science.gov (United States)

    Walenta, Stefan; Mueller-Klieser, Wolfgang

    2016-01-01

    This review is focused on the radiobiology of carbon ions compared to X-rays using multicellular models of tumors and normal mucosa. The first part summarizes basic radiobiological effects, as observed in cancer cells. The second, more clinically oriented part of the review, deals with radiation-induced cell migration and mucositis. Multicellular spheroids from V79 hamster cells were irradiated with X-rays or carbon ions under ambient or restricted oxygen supply conditions. Reliable oxygen enhancement ratios could be derived to be 2.9, 2.8, and 1.4 for irradiation with photons, 12C+6 in the plateau region, and 12C+6 in the Bragg peak, respectively. Similarly, a relative biological effectiveness of 4.3 and 2.1 for ambient pO2 and hypoxia was obtained, respectively. The high effectiveness of carbon ions was reflected by an enhanced accumulation of cells in G2/M and a dose-dependent massive induction of apoptosis. These data clearly show that heavy charged particles are more efficient in sterilizing tumor cells than conventional irradiation even under hypoxic conditions. Clinically relevant doses (3 Gy) of X-rays induced an increase in migratory activity of U87 but not of LN229 or HCT116 tumor cells. Such an increase in cell motility following irradiation in situ could be the source of recurrence. In contrast, carbon ion treatment was associated with a dose-dependent decrease in migration with all cell lines and under all conditions investigated. The radiation-induced loss of cell motility was correlated, in most cases, with corresponding changes in β1 integrin expression. The photon-induced increase in cell migration was paralleled by an elevated phosphorylation status of the epidermal growth factor receptor and AKT-ERK1/2 pathway. Such a hyperphosphorylation did not occur during 12C+6 irradiation under all conditions registered. Comparing the gene toxicity of X-rays with that of particles using the γH2AX technique in organotypic cultures of the oral mucosa, the

  7. Differential Superiority of Heavy Charged-Particle Irradiation to X-Rays: Studies on Biological Effectiveness and Side Effect Mechanisms in Multicellular Tumor and Normal Tissue Models.

    Science.gov (United States)

    Walenta, Stefan; Mueller-Klieser, Wolfgang

    2016-01-01

    This review is focused on the radiobiology of carbon ions compared to X-rays using multicellular models of tumors and normal mucosa. The first part summarizes basic radiobiological effects, as observed in cancer cells. The second, more clinically oriented part of the review, deals with radiation-induced cell migration and mucositis. Multicellular spheroids from V79 hamster cells were irradiated with X-rays or carbon ions under ambient or restricted oxygen supply conditions. Reliable oxygen enhancement ratios could be derived to be 2.9, 2.8, and 1.4 for irradiation with photons, (12)C(+6) in the plateau region, and (12)C(+6) in the Bragg peak, respectively. Similarly, a relative biological effectiveness of 4.3 and 2.1 for ambient pO2 and hypoxia was obtained, respectively. The high effectiveness of carbon ions was reflected by an enhanced accumulation of cells in G2/M and a dose-dependent massive induction of apoptosis. These data clearly show that heavy charged particles are more efficient in sterilizing tumor cells than conventional irradiation even under hypoxic conditions. Clinically relevant doses (3 Gy) of X-rays induced an increase in migratory activity of U87 but not of LN229 or HCT116 tumor cells. Such an increase in cell motility following irradiation in situ could be the source of recurrence. In contrast, carbon ion treatment was associated with a dose-dependent decrease in migration with all cell lines and under all conditions investigated. The radiation-induced loss of cell motility was correlated, in most cases, with corresponding changes in β1 integrin expression. The photon-induced increase in cell migration was paralleled by an elevated phosphorylation status of the epidermal growth factor receptor and AKT-ERK1/2 pathway. Such a hyperphosphorylation did not occur during (12)C(+6) irradiation under all conditions registered. Comparing the gene toxicity of X-rays with that of particles using the γH2AX technique in organotypic cultures of the oral

  8. Scattering of Gaussian beam by a spherical particle with a spheroidal inclusion

    International Nuclear Information System (INIS)

    Zhang Huayong; Liao Tongqing

    2011-01-01

    A generalized Lorenz-Mie theory framework (GLMT) is applied to the study of Gaussian beam scattering by a spherical particle with an embedded spheroid at the center. By virtue of a transformation between the spherical and spheroidal vector wave functions, a theoretical procedure is developed to deal with the boundary conditions. Numerical results of the normalized differential scattering cross section are presented.

  9. Spheroidal corrections to the spherical and parabolic bases of the hydrogen atom

    International Nuclear Information System (INIS)

    Mardyan, L.G.; Pogosyan, G.S.; Sisakyan, A.N.

    1986-01-01

    This paper introduces the bases of the hydrogen atom and obtains recursion relations that determine the expansion of the spheroidal basis with respect to its parabolic basis. The leading spheroidal corrections to the spherical and parabolic bases are calculated by perturbation theory

  10. Stability of self-gravitating homogeneous spheroid with azimuthal magnetic field. I

    International Nuclear Information System (INIS)

    Antonov, V.A.; Zheleznyak, O.A.

    1988-01-01

    The influence of a frozen magnetic field on the stability of a self-gravitating homogeneous spheroid with respect to a deformation that transforms it into a triaxial ellipsoid is investigated. It is shown that an azimuthal magnetic field is a stabilizing factor, allowing the spheroid to be stable at e > e/sub cr/ = 0.95285

  11. Multiplexing spheroid volume, resazurin and acid phosphatase viability assays for high-throughput screening of tumour spheroids and stem cell neurospheres.

    Directory of Open Access Journals (Sweden)

    Delyan P Ivanov

    Full Text Available Three-dimensional cell culture has many advantages over monolayer cultures, and spheroids have been hailed as the best current representation of small avascular tumours in vitro. However their adoption in regular screening programs has been hindered by uneven culture growth, poor reproducibility and lack of high-throughput analysis methods for 3D. The objective of this study was to develop a method for a quick and reliable anticancer drug screen in 3D for tumour and human foetal brain tissue in order to investigate drug effectiveness and selective cytotoxic effects. Commercially available ultra-low attachment 96-well round-bottom plates were employed to culture spheroids in a rapid, reproducible manner amenable to automation. A set of three mechanistically different methods for spheroid health assessment (Spheroid volume, metabolic activity and acid phosphatase enzyme activity were validated against cell numbers in healthy and drug-treated spheroids. An automated open-source ImageJ macro was developed to enable high-throughput volume measurements. Although spheroid volume determination was superior to the other assays, multiplexing it with resazurin reduction and phosphatase activity produced a richer picture of spheroid condition. The ability to distinguish between effects on malignant and the proliferating component of normal brain was tested using etoposide on UW228-3 medulloblastoma cell line and human neural stem cells. At levels below 10 µM etoposide exhibited higher toxicity towards proliferating stem cells, whereas at concentrations above 10 µM the tumour spheroids were affected to a greater extent. The high-throughput assay procedures use ready-made plates, open-source software and are compatible with standard plate readers, therefore offering high predictive power with substantial savings in time and money.

  12. A theoretical study of the spheroidal droplet evaporation in forced convection

    Energy Technology Data Exchange (ETDEWEB)

    Li, Jie, E-mail: leejay1986@163.com; Zhang, Jian

    2014-11-07

    In many applications, the shape of a droplet may be assumed to be an oblate spheroid. A theoretical study is conducted on the evaporation of an oblate spheroidal droplet under forced convection conditions. Closed-form analytical expressions of the mass evaporation rate for an oblate spheroid are derived, in the regime of controlled mass-transfer and heat-transfer, respectively. The variation of droplet size during the evaporation process is presented in the regime of shrinking dynamic model. Comparing with the droplets having the same surface area, an increase in the aspect ratio enhances the mass evaporation rate and prolongs the burnout time. - Highlights: • Fully algebraic solutions for the spheroidal droplet evaporation rate is obtained. • We examine the effect of aspect ratio on the droplet evaporation. • We propose a calculation method of Nusselt number for spheroidal droplet.

  13. Creation of Cardiac Tissue Exhibiting Mechanical Integration of Spheroids Using 3D Bioprinting.

    Science.gov (United States)

    Ong, Chin Siang; Fukunishi, Takuma; Nashed, Andrew; Blazeski, Adriana; Zhang, Huaitao; Hardy, Samantha; DiSilvestre, Deborah; Vricella, Luca; Conte, John; Tung, Leslie; Tomaselli, Gordon; Hibino, Narutoshi

    2017-07-02

    This protocol describes 3D bioprinting of cardiac tissue without the use of biomaterials, using only cells. Cardiomyocytes, endothelial cells and fibroblasts are first isolated, counted and mixed at desired cell ratios. They are co-cultured in individual wells in ultra-low attachment 96-well plates. Within 3 days, beating spheroids form. These spheroids are then picked up by a nozzle using vacuum suction and assembled on a needle array using a 3D bioprinter. The spheroids are then allowed to fuse on the needle array. Three days after 3D bioprinting, the spheroids are removed as an intact patch, which is already spontaneously beating. 3D bioprinted cardiac patches exhibit mechanical integration of component spheroids and are highly promising in cardiac tissue regeneration and as 3D models of heart disease.

  14. A theoretical study of the spheroidal droplet evaporation in forced convection

    International Nuclear Information System (INIS)

    Li, Jie; Zhang, Jian

    2014-01-01

    In many applications, the shape of a droplet may be assumed to be an oblate spheroid. A theoretical study is conducted on the evaporation of an oblate spheroidal droplet under forced convection conditions. Closed-form analytical expressions of the mass evaporation rate for an oblate spheroid are derived, in the regime of controlled mass-transfer and heat-transfer, respectively. The variation of droplet size during the evaporation process is presented in the regime of shrinking dynamic model. Comparing with the droplets having the same surface area, an increase in the aspect ratio enhances the mass evaporation rate and prolongs the burnout time. - Highlights: • Fully algebraic solutions for the spheroidal droplet evaporation rate is obtained. • We examine the effect of aspect ratio on the droplet evaporation. • We propose a calculation method of Nusselt number for spheroidal droplet

  15. Bacterial stigmergy: an organising principle of multicellular collective behaviours of bacteria.

    Science.gov (United States)

    Gloag, Erin S; Turnbull, Lynne; Whitchurch, Cynthia B

    2015-01-01

    The self-organisation of collective behaviours often manifests as dramatic patterns of emergent large-scale order. This is true for relatively "simple" entities such as microbial communities and robot "swarms," through to more complex self-organised systems such as those displayed by social insects, migrating herds, and many human activities. The principle of stigmergy describes those self-organised phenomena that emerge as a consequence of indirect communication between individuals of the group through the generation of persistent cues in the environment. Interestingly, despite numerous examples of multicellular behaviours of bacteria, the principle of stigmergy has yet to become an accepted theoretical framework that describes how bacterial collectives self-organise. Here we review some examples of multicellular bacterial behaviours in the context of stigmergy with the aim of bringing this powerful and elegant self-organisation principle to the attention of the microbial research community.

  16. Neutron diffusion in spheroidal, bispherical, and toroidal systems

    International Nuclear Information System (INIS)

    Williams, M.M.R.

    1986-01-01

    The neutron flux has been studied around absorbing bodies of spheroidal, bispherical, and toroidal shapes in an infinite nonabsorbing medium. Exact solutions have been obtained by using effective boundary conditions at the surfaces of the absorbing bodies. The problems considered are as follows: 1. Neutron flux and current distributions around prolate and oblate spheroids. It is shown that an equivalent sphere approximation can lead to accurate values for the rate of absorption. 2. Neutron flux and current in a bispherical system of unequal spheres. Three separate situations arise here: (a) two absorbing spheres, (b) two spherical sources, and (c) one spherical source and one absorbing sphere. It is shown how the absorption rate in the two spheres depends on their separation. 3. Neutron flux and current in a toroidal system: (a) an absorbing toroid and (b) a toroidal source. The latter case simulates the flux distribution from a thermonuclear reactor vessel. Finally, a brief description of how these techniques can be extended to multiregion problems is given

  17. Two step continuous method to synthesize colloidal spheroid gold nanorods.

    Science.gov (United States)

    Chandra, S; Doran, J; McCormack, S J

    2015-12-01

    This research investigated a two-step continuous process to synthesize colloidal suspension of spheroid gold nanorods. In the first step; gold precursor was reduced to seed-like particles in the presence of polyvinylpyrrolidone and ascorbic acid. In continuous second step; silver nitrate and alkaline sodium hydroxide produced various shape and size Au nanoparticles. The shape was manipulated through weight ratio of ascorbic acid to silver nitrate by varying silver nitrate concentration. The specific weight ratio of 1.35-1.75 grew spheroid gold nanorods of aspect ratio ∼1.85 to ∼2.2. Lower weight ratio of 0.5-1.1 formed spherical nanoparticle. The alkaline medium increased the yield of gold nanorods and reduced reaction time at room temperature. The synthesized gold nanorods retained their shape and size in ethanol. The surface plasmon resonance was red shifted by ∼5 nm due to higher refractive index of ethanol than water. Copyright © 2015 Elsevier Inc. All rights reserved.

  18. Sponge cell reaggregation: Cellular structure and morphogenetic potencies of multicellular aggregates.

    Science.gov (United States)

    Lavrov, Andrey I; Kosevich, Igor A

    2016-02-01

    Sponges (phylum Porifera) are one of the most ancient extant multicellular animals and can provide valuable insights into origin and early evolution of Metazoa. High plasticity of cell differentiations and anatomical structure is characteristic feature of sponges. Present study deals with sponge cell reaggregation after dissociation as the most outstanding case of sponge plasticity. Dynamic of cell reaggregation and structure of multicellular aggregates of three demosponge species (Halichondria panicea (Pallas, 1766), Haliclona aquaeductus (Sсhmidt, 1862), and Halisarca dujardinii Johnston, 1842) were studied. Sponge tissue dissociation was performed mechanically. Resulting cell suspensions were cultured at 8-10°C for at least 5 days. Structure of multicellular aggregates was studied by light, transmission and scanning electron microscopy. Studied species share common stages of cell reaggregation-primary multicellular aggregates, early-stage primmorphs and primmorphs, but the rate of reaggregation varies considerably among species. Only cells of H. dujardinii are able to reconstruct functional and viable sponge after primmorphs formation. Sponge reconstruction in this species occurs due to active cell locomotion. Development of H. aquaeductus and H. panicea cells ceases at the stages of early primmorphs and primmorphs, respectively. Development of aggregates of these species is most likely arrested due to immobility of the majority of cells inside them. However, the inability of certain sponge species to reconstruct functional and viable individuals during cell reaggregation may be not a permanent species-specific characteristic, but depends on various factors, including the stage of the life cycle and experimental conditions. © 2016 Wiley Periodicals, Inc.

  19. Processing and characterization of multi-cellular monolithic bioceramics for bone regenerative scaffolds

    International Nuclear Information System (INIS)

    Ari-Wahjoedi, Bambang; Ginta, Turnad Lenggo; Parman, Setyamartana; Abustaman, Mohd Zikri Ahmad

    2014-01-01

    Multicellular monolithic ceramic body is a ceramic material which has many gas or liquid passages partitioned by thin walls throughout the bulk material. There are many currently known advanced industrial applications of multicellular ceramics structures i.e. as supports for various catalysts, electrode support structure for solid oxide fuel cells, refractories, electric/electronic materials, aerospace vehicle re-entry heat shields and biomaterials for dental as well as orthopaedic implants by naming only a few. Multicellular ceramic bodies are usually made of ceramic phases such as mullite, cordierite, aluminum titanate or pure oxides such as silica, zirconia and alumina. What make alumina ceramics is excellent for the above functions are the intrinsic properties of alumina which are hard, wear resistant, excellent dielectric properties, resists strong acid and alkali attacks at elevated temperatures, good thermal conductivities, high strength and stiffness as well as biocompatible. In this work the processing technology leading to truly multicellular monolithic alumina ceramic bodies and their characterization are reported. Ceramic slip with 66 wt.% solid loading was found to be optimum as impregnant to the polyurethane foam template. Mullitic ceramic composite of alumina-sodium alumino disilicate-Leucite-like phases with bulk and true densities of 0.852 and 1.241 g cm −3 respectively, pore linear density of ±35 cm −1 , linear and bulk volume shrinkages of 7-16% and 32 vol.% were obtained. The compressive strength and elastic modulus of the bioceramics are ≈0.5-1.0 and ≈20 MPa respectively

  20. Processing and characterization of multi-cellular monolithic bioceramics for bone regenerative scaffolds

    Energy Technology Data Exchange (ETDEWEB)

    Ari-Wahjoedi, Bambang, E-mail: bambang-ariwahjoedi@petronas.com.my [Department of Fundamental and Applied Sciences, Universiti Teknologi PETRONAS, Bandar Seri Iskandar, 31750 Tronoh, Perak Darul Ridzuan (Malaysia); Centre for Intelligent Signal and Imaging Research, Universiti Teknologi PETRONAS, Bandar Seri Iskandar (Malaysia); Ginta, Turnad Lenggo [Department of Mechanical Engineering, Universiti Teknologi PETRONAS, Bandar Seri Iskandar, 31750 Tronoh, Perak Darul Ridzuan (Malaysia); Centre for Intelligent Signal and Imaging Research, Universiti Teknologi PETRONAS, Bandar Seri Iskandar, 31750 Tro (Malaysia); Parman, Setyamartana [Department of Mechanical Engineering, Universiti Teknologi PETRONAS, Bandar Seri Iskandar, 31750 Tronoh, Perak Darul Ridzuan (Malaysia); Abustaman, Mohd Zikri Ahmad [Kebabangan Petroleum Operating Company Sdn Bhd, Lvl. 52, Tower 2, PETRONAS Twin Towers, KLCC, 50088 Kuala Lumpur (Malaysia)

    2014-10-24

    Multicellular monolithic ceramic body is a ceramic material which has many gas or liquid passages partitioned by thin walls throughout the bulk material. There are many currently known advanced industrial applications of multicellular ceramics structures i.e. as supports for various catalysts, electrode support structure for solid oxide fuel cells, refractories, electric/electronic materials, aerospace vehicle re-entry heat shields and biomaterials for dental as well as orthopaedic implants by naming only a few. Multicellular ceramic bodies are usually made of ceramic phases such as mullite, cordierite, aluminum titanate or pure oxides such as silica, zirconia and alumina. What make alumina ceramics is excellent for the above functions are the intrinsic properties of alumina which are hard, wear resistant, excellent dielectric properties, resists strong acid and alkali attacks at elevated temperatures, good thermal conductivities, high strength and stiffness as well as biocompatible. In this work the processing technology leading to truly multicellular monolithic alumina ceramic bodies and their characterization are reported. Ceramic slip with 66 wt.% solid loading was found to be optimum as impregnant to the polyurethane foam template. Mullitic ceramic composite of alumina-sodium alumino disilicate-Leucite-like phases with bulk and true densities of 0.852 and 1.241 g cm{sup −3} respectively, pore linear density of ±35 cm{sup −1}, linear and bulk volume shrinkages of 7-16% and 32 vol.% were obtained. The compressive strength and elastic modulus of the bioceramics are ≈0.5-1.0 and ≈20 MPa respectively.

  1. Cell motility dynamics: a novel segmentation algorithm to quantify multi-cellular bright field microscopy images.

    Directory of Open Access Journals (Sweden)

    Assaf Zaritsky

    Full Text Available Confocal microscopy analysis of fluorescence and morphology is becoming the standard tool in cell biology and molecular imaging. Accurate quantification algorithms are required to enhance the understanding of different biological phenomena. We present a novel approach based on image-segmentation of multi-cellular regions in bright field images demonstrating enhanced quantitative analyses and better understanding of cell motility. We present MultiCellSeg, a segmentation algorithm to separate between multi-cellular and background regions for bright field images, which is based on classification of local patches within an image: a cascade of Support Vector Machines (SVMs is applied using basic image features. Post processing includes additional classification and graph-cut segmentation to reclassify erroneous regions and refine the segmentation. This approach leads to a parameter-free and robust algorithm. Comparison to an alternative algorithm on wound healing assay images demonstrates its superiority. The proposed approach was used to evaluate common cell migration models such as wound healing and scatter assay. It was applied to quantify the acceleration effect of Hepatocyte growth factor/scatter factor (HGF/SF on healing rate in a time lapse confocal microscopy wound healing assay and demonstrated that the healing rate is linear in both treated and untreated cells, and that HGF/SF accelerates the healing rate by approximately two-fold. A novel fully automated, accurate, zero-parameters method to classify and score scatter-assay images was developed and demonstrated that multi-cellular texture is an excellent descriptor to measure HGF/SF-induced cell scattering. We show that exploitation of textural information from differential interference contrast (DIC images on the multi-cellular level can prove beneficial for the analyses of wound healing and scatter assays. The proposed approach is generic and can be used alone or alongside traditional

  2. Cell motility dynamics: a novel segmentation algorithm to quantify multi-cellular bright field microscopy images.

    Science.gov (United States)

    Zaritsky, Assaf; Natan, Sari; Horev, Judith; Hecht, Inbal; Wolf, Lior; Ben-Jacob, Eshel; Tsarfaty, Ilan

    2011-01-01

    Confocal microscopy analysis of fluorescence and morphology is becoming the standard tool in cell biology and molecular imaging. Accurate quantification algorithms are required to enhance the understanding of different biological phenomena. We present a novel approach based on image-segmentation of multi-cellular regions in bright field images demonstrating enhanced quantitative analyses and better understanding of cell motility. We present MultiCellSeg, a segmentation algorithm to separate between multi-cellular and background regions for bright field images, which is based on classification of local patches within an image: a cascade of Support Vector Machines (SVMs) is applied using basic image features. Post processing includes additional classification and graph-cut segmentation to reclassify erroneous regions and refine the segmentation. This approach leads to a parameter-free and robust algorithm. Comparison to an alternative algorithm on wound healing assay images demonstrates its superiority. The proposed approach was used to evaluate common cell migration models such as wound healing and scatter assay. It was applied to quantify the acceleration effect of Hepatocyte growth factor/scatter factor (HGF/SF) on healing rate in a time lapse confocal microscopy wound healing assay and demonstrated that the healing rate is linear in both treated and untreated cells, and that HGF/SF accelerates the healing rate by approximately two-fold. A novel fully automated, accurate, zero-parameters method to classify and score scatter-assay images was developed and demonstrated that multi-cellular texture is an excellent descriptor to measure HGF/SF-induced cell scattering. We show that exploitation of textural information from differential interference contrast (DIC) images on the multi-cellular level can prove beneficial for the analyses of wound healing and scatter assays. The proposed approach is generic and can be used alone or alongside traditional fluorescence single

  3. Genomic analysis of organismal complexity in the multicellular green alga Volvox carteri

    Energy Technology Data Exchange (ETDEWEB)

    Prochnik, Simon E.; Umen, James; Nedelcu, Aurora; Hallmann, Armin; Miller, Stephen M.; Nishii, Ichiro; Ferris, Patrick; Kuo, Alan; Mitros, Therese; Fritz-Laylin, Lillian K.; Hellsten, Uffe; Chapman, Jarrod; Simakov, Oleg; Rensing, Stefan A.; Terry, Astrid; Pangilinan, Jasmyn; Kapitonov, Vladimir; Jurka, Jerzy; Salamov, Asaf; Shapiro, Harris; Schmutz, Jeremy; Grimwood, Jane; Lindquist, Erika; Lucas, Susan; Grigoriev, Igor V.; Schmitt, Rudiger; Kirk, David; Rokhsar, Daniel S.

    2010-07-01

    Analysis of the Volvox carteri genome reveals that this green alga's increased organismal complexity and multicellularity are associated with modifications in protein families shared with its unicellular ancestor, and not with large-scale innovations in protein coding capacity. The multicellular green alga Volvox carteri and its morphologically diverse close relatives (the volvocine algae) are uniquely suited for investigating the evolution of multicellularity and development. We sequenced the 138 Mb genome of V. carteri and compared its {approx}14,500 predicted proteins to those of its unicellular relative, Chlamydomonas reinhardtii. Despite fundamental differences in organismal complexity and life history, the two species have similar protein-coding potentials, and few species-specific protein-coding gene predictions. Interestingly, volvocine algal-specific proteins are enriched in Volvox, including those associated with an expanded and highly compartmentalized extracellular matrix. Our analysis shows that increases in organismal complexity can be associated with modifications of lineage-specific proteins rather than large-scale invention of protein-coding capacity.

  4. On the origin of biological construction, with a focus on multicellularity.

    Science.gov (United States)

    van Gestel, Jordi; Tarnita, Corina E

    2017-10-17

    Biology is marked by a hierarchical organization: all life consists of cells; in some cases, these cells assemble into groups, such as endosymbionts or multicellular organisms; in turn, multicellular organisms sometimes assemble into yet other groups, such as primate societies or ant colonies. The construction of new organizational layers results from hierarchical evolutionary transitions, in which biological units (e.g., cells) form groups that evolve into new units of biological organization (e.g., multicellular organisms). Despite considerable advances, there is no bottom-up, dynamical account of how, starting from the solitary ancestor, the first groups originate and subsequently evolve the organizing principles that qualify them as new units. Guided by six central questions, we propose an integrative bottom-up approach for studying the dynamics underlying hierarchical evolutionary transitions, which builds on and synthesizes existing knowledge. This approach highlights the crucial role of the ecology and development of the solitary ancestor in the emergence and subsequent evolution of groups, and it stresses the paramount importance of the life cycle: only by evaluating groups in the context of their life cycle can we unravel the evolutionary trajectory of hierarchical transitions. These insights also provide a starting point for understanding the types of subsequent organizational complexity. The central research questions outlined here naturally link existing research programs on biological construction (e.g., on cooperation, multilevel selection, self-organization, and development) and thereby help integrate knowledge stemming from diverse fields of biology.

  5. UVES Abundances of Stars in Nearby Dwarf Spheroidal Galaxies

    Science.gov (United States)

    Tolstoy, Eline; Venn, Kim; Shetrone, Matt; Primas, Francesca; Hill, Vanessa; Kaufer, Andreas; Szeifert, Thomas

    2002-07-01

    It is a truth universally acknowledged, that a galaxy in possession of a good quantity of gas must want to form stars. It is the details of how and why that baffle us all. The simplest theories either would have this process a carefully self-regulated affair, or one that goes completely out of control and is capable of wrecking the galaxy which hosts it. Of course the majority of galaxies seem to amble along somewhere between these two extremes, and the mean properties tend to favour a quiescent self-regulated evolutionary scenario. But there area variety of observations which require us to invoke transitory ‘bursts’ of star-formation at one time or another in most galaxy types. Several nearby dwarf spheroidal galaxies have clearly determined star-formation histories with apparent periods of zero star formation followed by periods of fairly active star formation. If we are able to understand what separated these bursts we would understand several important phenomena in galaxy evolution. Were these galaxies able to clear out their gas reservoir in a burst of star formation? How did this gas return? or did it? Have these galaxies receieved gas from the IGM instead? Could stars from these types of galaxy contribute significantly to the halo population in our Galaxy? To answer these questions we need to combine accurate stellar photometry and Colour-Magnitude Diagram interpretation with detailed metal abundances to combine a star-formation rate versus time with a range of element abundances with time. Different elements trace different evolutionary process (e.g., relative contributions of type I and II supernovae). We often aren't even sure of the abundance spread in these galaxies. We have collected detailed high resolution UVES spectra of four nearby dwarf spheroidal galaxies (Sculptor, Fornax, Leo I & Carina) to begin to answer these questions. This is a precursor study to a more complete study with FLAMES. We presented at this meeting the initial results for

  6. Proteomic approach toward molecular backgrounds of drug resistance of osteosarcoma cells in spheroid culture system.

    Science.gov (United States)

    Arai, Kazuya; Sakamoto, Ruriko; Kubota, Daisuke; Kondo, Tadashi

    2013-08-01

    Chemoresistance is one of the most critical prognostic factors in osteosarcoma, and elucidation of the molecular backgrounds of chemoresistance may lead to better clinical outcomes. Spheroid cells resemble in vivo cells and are considered an in vitro model for the drug discovery. We found that spheroid cells displayed more chemoresistance than conventional monolayer cells across 11 osteosarcoma cell lines. To investigate the molecular mechanisms underlying the resistance to chemotherapy, we examined the proteomic differences between the monolayer and spheroid cells by 2D-DIGE. Of the 4762 protein species observed, we further investigated 435 species with annotated mass spectra in the public proteome database, Genome Medicine Database of Japan Proteomics. Among the 435 protein species, we found that 17 species exhibited expression level differences when the cells formed spheroids in more than five cell lines and four species out of these 17 were associated with spheroid-formation associated resistance to doxorubicin. We confirmed the upregulation of cathepsin D in spheroid cells by western blotting. Cathepsin D has been implicated in chemoresistance of various malignancies but has not previously been implemented in osteosarcoma. Our study suggested that the spheroid system may be a useful tool to reveal the molecular backgrounds of chemoresistance in osteosarcoma. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  7. Detachably assembled microfluidic device for perfusion culture and post-culture analysis of a spheroid array.

    Science.gov (United States)

    Sakai, Yusuke; Hattori, Koji; Yanagawa, Fumiki; Sugiura, Shinji; Kanamori, Toshiyuki; Nakazawa, Kohji

    2014-07-01

    Microfluidic devices permit perfusion culture of three-dimensional (3D) tissue, mimicking the flow of blood in vascularized 3D tissue in our body. Here, we report a microfluidic device composed of a two-part microfluidic chamber chip and multi-microwell array chip able to be disassembled at the culture endpoint. Within the microfluidic chamber, an array of 3D tissue aggregates (spheroids) can be formed and cultured under perfusion. Subsequently, detailed post-culture analysis of the spheroids collected from the disassembled device can be performed. This device facilitates uniform spheroid formation, growth analysis in a high-throughput format, controlled proliferation via perfusion flow rate, and post-culture analysis of spheroids. We used the device to culture spheroids of human hepatocellular carcinoma (HepG2) cells under two controlled perfusion flow rates. HepG2 spheroids exhibited greater cell growth at higher perfusion flow rates than at lower perfusion flow rates, and exhibited different metabolic activity and mRNA and protein expression under the different flow rate conditions. These results show the potential of perfusion culture to precisely control the culture environment in microfluidic devices. The construction of spheroid array chambers allows multiple culture conditions to be tested simultaneously, with potential applications in toxicity and drug screening. Copyright © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  8. An injectable spheroid system with genetic modification for cell transplantation therapy.

    Science.gov (United States)

    Uchida, Satoshi; Itaka, Keiji; Nomoto, Takahiro; Endo, Taisuke; Matsumoto, Yu; Ishii, Takehiko; Kataoka, Kazunori

    2014-03-01

    The new methodology to increase a therapeutic potential of cell transplantation was developed here by the use of three-dimensional spheroids of transplanting cells subsequent to the genetic modification with non-viral DNA vectors, polyplex nanomicelles. Particularly, spheroids in regulated size of 100-μm of primary hepatocytes transfected with luciferase gene were formed on the micropatterned culture plates coated with thermosensitive polymer, and were recovered in the form of injectable liquid suspension simply by cooling the plates. After subcutaneously transplanting these hepatocyte spheroids, efficient transgene expression was observed in host tissue for more than a month, whereas transplantation of a single-cell suspension from a monolayer culture resulted in an only transient expression. The spheroid system contributed to the preservation of innate functions of transplanted hepatocytes in the host tissue, such as albumin expression, thereby possessing high potential for expressing transgene. Intravital observation of transplanted cells showed that those from spheroid cultures had a tendency to localize in the vicinity of blood vessels, making a favorable microenvironment for preserving cell functionality. Furthermore, spheroids transfected with erythropoietin-expressing DNA showed a significantly higher hematopoietic effect than that of cell suspensions from monolayer cultures, demonstrating high potential of this genetically-modified spheroid transplantation system for therapeutic applications. Copyright © 2013 Elsevier Ltd. All rights reserved.

  9. Identification of Lgr5-Independent Spheroid-Generating Progenitors of the Mouse Fetal Intestinal Epithelium

    Directory of Open Access Journals (Sweden)

    Roxana C. Mustata

    2013-10-01

    Full Text Available Immortal spheroids were generated from fetal mouse intestine using the culture system initially developed to culture organoids from adult intestinal epithelium. Spheroid proportion progressively decreases from fetal to postnatal period, with a corresponding increase in production of organoids. Like organoids, spheroids show Wnt-dependent indefinite self-renewing properties but display a poorly differentiated phenotype reminiscent of incompletely caudalized progenitors. The spheroid transcriptome is strikingly different from that of adult intestinal stem cells, with minimal overlap of Wnt target gene expression. The receptor LGR4, but not LGR5, is essential for their growth. Trop2/Tacstd2 and Cnx43/Gja1, two markers highly enriched in spheroids, are expressed throughout the embryonic-day-14 intestinal epithelium. Comparison of in utero and neonatal lineage tracing using Cnx43-CreER and Lgr5-CreERT2 mice identified spheroid-generating cells as developmental progenitors involved in generation of the prenatal intestinal epithelium. Ex vivo, spheroid cells have the potential to differentiate into organoids, qualifying as a fetal type of intestinal stem cell.

  10. Promotion of malignant phenotype after disruption of the three-dimensional structure of cultured spheroids from colorectal cancer.

    Science.gov (United States)

    Piulats, Jose M; Kondo, Jumpei; Endo, Hiroko; Ono, Hiromasa; Hagihara, Takeshi; Okuyama, Hiroaki; Nishizawa, Yasuko; Tomita, Yasuhiko; Ohue, Masayuki; Okita, Kouki; Oyama, Hidejiro; Bono, Hidemasa; Masuko, Takashi; Inoue, Masahiro

    2018-03-23

    Individual and small clusters of cancer cells may detach from the edges of a main tumor and invade vessels, which can act as the origin of metastasis; however, the mechanism for this phenomenon is not well understood. Using cancer tissue-originated spheroids, we studied whether disturbing the 3D architecture of cancer spheroids can provoke the reformation process and progression of malignancy. We developed a mechanical disruption method to achieve homogenous disruption of the spheroids while maintaining cell-cell contact. After the disruption, 9 spheroid lines from 9 patient samples reformed within a few hours, and 3 of the 9 lines exhibited accelerated spheroid growth. Marker expression, spheroid forming capacity, and tumorigenesis indicated that stemness increased after spheroid disruption. In addition, the spheroid forming capacity increased in 6 of 11 spheroid lines. The disruption signature determined by gene expression profiling supported the incidence of remodeling and predicted the prognosis of patients with colorectal cancer. Furthermore, WNT and HER3 signaling were increased in the reformed spheroids, and suppression of these signaling pathways attenuated the increased proliferation and stemness after the disruption. Overall, the disruption and subsequent reformation of cancer spheroids promoted malignancy-related phenotypes through the activation of the WNT and ERBB pathways.

  11. Pervasive plastisphere: First record of plastics in egagropiles (Posidonia spheroids).

    Science.gov (United States)

    Pietrelli, Loris; Di Gennaro, Alessia; Menegoni, Patrizia; Lecce, Francesca; Poeta, Gianluca; Acosta, Alicia T R; Battisti, Corrado; Iannilli, Valentina

    2017-10-01

    The ability of Posidonia oceanica spheroids (egagropiles, EG) to incorporate plastics was investigated along the central Italy coast. Plastics were found in the 52.84% of the egagropiles collected (n = 685). The more represented size of plastics has range within 1-1.5 cm, comparable to the size of natural fibres. Comparing plastics occurring both in EG and in surrounding sand, Polyethylene, Polyester and Nylon were the most abundant polymers in EG, while PSE, PE, PP and PET were the most represented in sand. In particular PE and PP were significantly more represented in sand, while PE, Nylon, Polyester and microfibers (as pills) were more represented in EG. Within plastics found in EG, 26.9% were microfibers as small pills (PET mixing. These microfibers might be produced by discharges from washing machines and currently represents an emerging pollutant with widespread distribution in marine and freshwater ecosystems. Copyright © 2017 Elsevier Ltd. All rights reserved.

  12. Modes of spheroidal ion plasmas at the Brillouin limit

    International Nuclear Information System (INIS)

    Tinkle, M.D.; Greaves, R.G.; Surko, C.M.

    1996-01-01

    The confinement properties and collective modes of single-component plasmas are investigated in a quadrupole Penning trap. Brillouin-density pure ion plasmas are generated by electron-beam ionization of a low-pressure gas. Large, spheroidal, steady-state plasmas are produced, extending out to contact one or more of the trap electrodes. With the density fixed at the Brillouin limit by the high ion production rate, the electrode potentials determine the plasma shape. The frequencies of azimuthally propagating cyclotron and diocotron modes are found to vary significantly with the plasma aspect ratio. For oblate plasmas, the frequencies are in good agreement with a simple fluid model. copyright 1996 American Institute of Physics

  13. Numerical simulations of bistable flows in precessing spheroidal shells

    Science.gov (United States)

    Vormann, J.; Hansen, U.

    2018-05-01

    Precession of the rotation axis is an often neglected mechanical driving mechanism for flows in planetary interiors, through viscous coupling at the boundaries and topographic forcing in non-spherical geometries. We investigate precession-driven flows in spheroidal shells over a wide range of parameters and test the results against theoretical predictions. For Ekman numbers down to 8.0 × 10-7, we see a good accordance with the work of Busse, who assumed the precession-driven flow to be dominated by a rigid rotation component that is tilted to the main rotation axis. The velocity fields show localized small-scale structures for lower Ekman numbers and clear signals of inertial waves for some parameters. For the case of moderate viscosity and strong deformation, we report the realization of multiple solutions at the same parameter combination, depending on the initial condition.

  14. Patient-specific three-dimensional explant spheroids derived from human nasal airway epithelium

    DEFF Research Database (Denmark)

    Marthin, June Kehlet; Stevens, Elizabeth Munkebjerg; Larsen, Lars Allan

    2017-01-01

    BACKGROUND: Three-dimensional explant spheroid formation is an ex vivo technique previously used in studies of airway epithelial ion and water transport. Explanted cells and sheets of nasal epithelium form fully differentiated spheroids enclosing a partly fluid-filled lumen with the ciliated apical...... surface facing the outside and accessible for analysis of ciliary function. METHODS: We performed a two-group comparison study of ciliary beat pattern and ciliary beat frequency in spheroids derived from nasal airway epithelium in patients with primary ciliary dyskinesia (PCD) and in healthy controls...... in the investigation of pathophysiological aspects and drug effects in human nasal airway epithelium....

  15. Scattering of a high-order Bessel beam by a spheroidal particle

    Science.gov (United States)

    Han, Lu

    2018-05-01

    Within the framework of generalized Lorenz-Mie theory (GLMT), scattering from a homogeneous spheroidal particle illuminated by a high-order Bessel beam is formulated analytically. The high-order Bessel beam is expanded in terms of spheroidal vector wave functions, where the spheroidal beam shape coefficients (BSCs) are computed conveniently using an intrinsic method. Numerical results concerning scattered field in the far zone are displayed for various parameters of the incident Bessel beam and of the scatter. These results are expected to provide useful insights into the scattering of a Bessel beam by nonspherical particles and particle manipulation applications using Bessel beams.

  16. A theoretical study of hot plasma spheroids in the presence of low-frequency electromagnetic waves

    Science.gov (United States)

    Ahmadizadeh, Y.; Jazi, B.; Barjesteh, S.

    2016-07-01

    While taking into account thermal motion of electrons, scattering of electromagnetic waves with low frequency from hot plasma spheroids is investigated. In this theoretical research, ions are heavy to respond to electromagnetic fluctuations. The solution of scalar wave equation in spheroidal coordinates for electric potential inside the plasma spheroids are obtained. The variations of resonance frequencies vs. Debye length are studied and consistency between the obtained results in this paper and the results for the well-known plasma objects such as plasma column and spherical plasma have been proved.

  17. Low-temperature plasma spheroidizing of polydisperse powders of refractory materials

    International Nuclear Information System (INIS)

    Tsymbalist, M.M.; Rudenskaya, N.A.; Kuz'min, B.P.; Pan'kov, V.A.

    2003-01-01

    A model is developed for heating and melting of a spherical particle, when powder processing in low temperature plasma, with the aim of estimation of the dependence of the degree of fusion on particle size for various materials. Spheroidizing of various refractory material powders close in shape and size composition is experimentally performed. Experimental and calculation estimates of spheroidizing criteria for the materials studied are in a satisfactory agreement. The influence of basic physical properties of refractory materials and plasma processing parameters on the degree of particle spheroidizing is analyzed [ru

  18. Tensile Forces Originating from Cancer Spheroids Facilitate Tumor Invasion.

    Directory of Open Access Journals (Sweden)

    Katarzyna S Kopanska

    Full Text Available The mechanical properties of tumors and the tumor environment provide important information for the progression and characterization of cancer. Tumors are surrounded by an extracellular matrix (ECM dominated by collagen I. The geometrical and mechanical properties of the ECM play an important role for the initial step in the formation of metastasis, presented by the migration of malignant cells towards new settlements as well as the vascular and lymphatic system. The extent of this cell invasion into the ECM is a key medical marker for cancer prognosis. In vivo studies reveal an increased stiffness and different architecture of tumor tissue when compared to its healthy counterparts. The observed parallel collagen organization on the tumor border and radial arrangement at the invasion zone has raised the question about the mechanisms organizing these structures. Here we study the effect of contractile forces originated from model tumor spheroids embedded in a biomimetic collagen I matrix. We show that contractile forces act immediately after seeding and deform the ECM, thus leading to tensile radial forces within the matrix. Relaxation of this tension via cutting the collagen does reduce invasion, showing a mechanical relation between the tensile state of the ECM and invasion. In turn, these results suggest that tensile forces in the ECM facilitate invasion. Furthermore, simultaneous contraction of the ECM and tumor growth leads to the condensation and reorientation of the collagen at the spheroid's surface. We propose a tension-based model to explain the collagen organization and the onset of invasion by forces originating from the tumor.

  19. Genome duplication and mutations in ACE2 cause multicellular, fast-sedimenting phenotypes in evolved Saccharomyces cerevisiae.

    Science.gov (United States)

    Oud, Bart; Guadalupe-Medina, Victor; Nijkamp, Jurgen F; de Ridder, Dick; Pronk, Jack T; van Maris, Antonius J A; Daran, Jean-Marc

    2013-11-05

    Laboratory evolution of the yeast Saccharomyces cerevisiae in bioreactor batch cultures yielded variants that grow as multicellular, fast-sedimenting clusters. Knowledge of the molecular basis of this phenomenon may contribute to the understanding of natural evolution of multicellularity and to manipulating cell sedimentation in laboratory and industrial applications of S. cerevisiae. Multicellular, fast-sedimenting lineages obtained from a haploid S. cerevisiae strain in two independent evolution experiments were analyzed by whole genome resequencing. The two evolved cell lines showed different frameshift mutations in a stretch of eight adenosines in ACE2, which encodes a transcriptional regulator involved in cell cycle control and mother-daughter cell separation. Introduction of the two ace2 mutant alleles into the haploid parental strain led to slow-sedimenting cell clusters that consisted of just a few cells, thus representing only a partial reconstruction of the evolved phenotype. In addition to single-nucleotide mutations, a whole-genome duplication event had occurred in both evolved multicellular strains. Construction of a diploid reference strain with two mutant ace2 alleles led to complete reconstruction of the multicellular-fast sedimenting phenotype. This study shows that whole-genome duplication and a frameshift mutation in ACE2 are sufficient to generate a fast-sedimenting, multicellular phenotype in S. cerevisiae. The nature of the ace2 mutations and their occurrence in two independent evolution experiments encompassing fewer than 500 generations of selective growth suggest that switching between unicellular and multicellular phenotypes may be relevant for competitiveness of S. cerevisiae in natural environments.

  20. Coupling gene expression and multicellular morphogenesis during fruiting body formation in Myxococcus xanthus

    DEFF Research Database (Denmark)

    Søgaard-Andersen, L.; Overgaard, M.; Lobedanz, S.

    2003-01-01

    xanthus illustrates this coupling in the construction of a multicellular structure. Fruiting body formation involves two stages: aggregation of cells into mounds and the position-specific sporulation of cells that have accumulated inside mounds. Developmental gene expression propels these two processes...... morphogenesis. Accumulation of the C-signal is tightly regulated and involves transcriptional activation of the csgA gene and proteolysis of the full-length CsgA protein to produce the shorter cell surface-associated 17 kDa C-signal protein. The C-signal induces aggregation, sporulation and developmental gene...

  1. The Paramyxea Levine 1979: An original example of evolution towards multicellularity

    Science.gov (United States)

    Desportes, Isabelle

    1984-03-01

    The Paramyxea are parasitic in marine invertebrates. Their development is a sporulation involving the differentiation within a stem cell of several sporonts which produce spores made of cells enclosed inside each other. Three genera are recognized according to the number of spores and sporal cells, and the taxonomic position of the host (Polychaeta, Mollusca, Crustacea). The Paramyxea exhibit both protistan and metazoan characters. Their nine singlets centrioles are observed in different Protoctists whereas the fact that their sporal cells acquire distinctive cytological features may be interpreted as an evolution towards multicellularity.

  2. The Role of Functional Amyloids in Multicellular Growth and Development of Gram-Positive Bacteria

    DEFF Research Database (Denmark)

    Dragoš, Anna; Kovács, Ákos T.; Claessen, Dennis

    2017-01-01

    Amyloid fibrils play pivotal roles in all domains of life. In bacteria, these fibrillar structures are often part of an extracellular matrix that surrounds the producing organism and thereby provides protection to harsh environmental conditions. Here, we discuss the role of amyloid fibrils...... in the two distant Gram-positive bacteria, Streptomyces coelicolor and Bacillus subtilis. We describe how amyloid fibrils contribute to a multitude of developmental processes in each of these systems, including multicellular growth and community development. Despite this variety of tasks, we know...... surprisingly little about how their assembly is organized to fulfill all these roles....

  3. [Self-organization in the ontogeny of multicellular organisms: a computer simulation].

    Science.gov (United States)

    Markov, M A; Markov, A V

    2011-01-01

    The progress in understanding the patterns of evolution of ontogeny is hindered by the fact that many features of ontogeny are counterintuitive (as well as the features of other processes related to self-organization, self-assembly, and spontaneous increase in complexity). The basic principle of ontogeny of multicellular organisms is that it is the process of self-assembly of ordered multicellular structures by means of coordinated behavior of many individual modules (cells), each of which follows the same set of"rules" encoded in the genome. These rules are based on the genetic regulatory networks. We hypothesize that many specific features of ontogeny that seem nontrivial or enigmatic are, in fact, the inevitable consequences of this basic principle. If so, they do not need special explanations. In order to verify this hypothesis, we developed the computer program "Evo-Devo" based on the above principle. The program is designed to model the self-assembly of ordered multicellular structures from an aggregation of dividing cells that originate from a single original cell (zygote). Each cell follows a set of rules of behavior ("genotype") that can be specified arbitrarily by the experimenter, and is the same for all cells in the embryo (each cell is programmed in exactly the same way as all other cells). It is not allowed to specify rules for groups of cells or for the whole embryo: only local rules that should be followed at the level of a single cell are possible. The analysis of phenotypic implementation of different genotypes revealed several features which are present in the ontogeny of real organisms and are regularly reproduced in the model. These include: inherent stochasticity; inescapable necessity of development of stabilizing adaptations based on negative feedback in order to decrease this stochasticity; equifinality (noise resistance) resulting from these adaptations; the ability of ontogeny to respond to major perturbations by generating new

  4. Involvement of bacterial migration in the development of complex multicellular structures in Pseudomonas aeruginosa biofilms

    DEFF Research Database (Denmark)

    Klausen, Mikkel; Aaes-Jorgensen, A.; Molin, Søren

    2003-01-01

    development, we have performed an investigation with time-lapse confocal laser scanning microscopy of biofilms formed by various combinations of colour-coded P. aeruginosa wild type and motility mutants. We show that mushroom-shaped multicellular structures in P. aeruginosa biofilms can form in a sequential...... process involving a non-motile bacterial subpopulation and a migrating bacterial subpopulation. The non-motile bacteria form the mushroom stalks by growth in certain foci of the biofilm. The migrating bacteria form the mushroom caps by climbing the stalks and aggregating on the tops in a process which...

  5. Numerical modeling of working of a multicellular proportional counter aimed to individual dosimetry of neutrons

    International Nuclear Information System (INIS)

    Bordy, J.M.; Barthe, J.; Boutruche, B.

    1993-01-01

    The use of a personal dosimeter imposes severe constraints, particularly for tension of polarization and tolerable dimensions. That why a numerical modeling of this detector working is an appreciable help for conception. It allows to determine quickly the influence of modification of different parameters (nature and pressure of gas, dimension of electrodes, dimension of channels, tension of polarization,...) without having to make new prototypes. The aim of this report is to give some numerical results got with a multicellular counter with a cylindrical geometry. 6 figs

  6. Study of human prostate spheroids treated with zinc using X-ray microfluorescence

    Energy Technology Data Exchange (ETDEWEB)

    Leitao, Roberta G.; Lopes, Ricardo T.; Pereira, Gabriela R., E-mail: roberta@lin.ufrj.br, E-mail: gpereira@metalmat.ufrj.br [Coordenacao dos Cursos de Pos-Graduacao em Engenharia (COPPE/UFRJ), Rio de Janeiro, RJ (Brazil); Santos, Carlos A.N., E-mail: cansantos.bio@gmail.com [Instituto Nacional de Metrologia, Qualidade e Tecnologia (DIPRO/INMETRO), Duque de Caxias, RJ (Brazil). Lab. de Biotecnologia; Palumbo Junior, Antonio; Nasciutti, Luiz E., E-mail: nasciutt@ufrj.br [Universidade Federal do Rio de Janeiro (ICB/CCS/UFRJ), Rio de Janeiro, RJ (Brazil). Lab. de Interacoes Celulares; Souza, Pedro A.V.R., E-mail: pedroaugustoreis@uol.com.br [Hospital Federal do Andarai (HFA), Rio de Janeiro, RJ (Brazil). Servico de Urologia; Anjos, Marcelino J., E-mail: marcelin@lin.ufrj.br [Universidade Estatual do Rio de Janeiro (UERJ), Rio de Janeiro, RJ (Brazil). Inst. de Fisica

    2013-07-01

    Spheroids cell culture is a useful technique for tissue engineering or regenerative medicine re-search, pharmacological and toxicological studies, and fundamental studies in cell biology. In this study, we investigated Zn distribution in cell spheroids in benign prostate hyperplasia (BPH) and prostate cancer (DU145) and analyzed the differences in the response to Zinc (0-150 μM) treatment. The measurements were performed in standard geometry of 45 deg incidence, exciting with a white beam and using an optical capillary with 20 μm diameter collimation in the XRF beam line at the Synchrotron Light National Laboratory (Campinas, Brazil). The results showed non-uniform distribution of Zn in all the spheroids analyzed. The differential response to zinc of DU145 and BPH cell spheroids suggests that zinc may have an important role in prostate cancer and BPH diagnosis. (author)

  7. Axonal Spheroid Accumulation In the Brainstem and Spinal Cord of A Young Angus Cow with Ataxia.

    Science.gov (United States)

    Hanshaw, D M; Finnie, J W; Manavis, J; Kessell, A E

    2015-08-01

    An 18-month-old Angus cow presented with rapidly developing ataxia and subsequently died. The finding of large numbers of axonal spheroids in brainstem nuclei and spinal cord grey matter, bilaterally symmetrical in distribution, was consistent with a histopathological diagnosis of neuroaxonal dystrophy (NAD). Most of the axonal swellings were immunopositive to amyloid precursor protein, suggesting that interruption to axonal flow was important in their genesis. The topographical distribution of axonal spheroids in the brain and spinal cord in this bovine case closely resembled that found in the ovine neurodegenerative disorder termed NAD, in which axonal swellings are the major pathological feature. This appears to be the first reported case of this type of NAD in cattle. The aetiology of the spheroidal aggregations in this case was not determined. There was no evidence from the case history or neuropathology to indicate whether the axonal spheroids in this case involved an acquired or heritable aetiology. © 2015 Australian Veterinary Association.

  8. Processes of Formation of Spheroidal Concretions and Inferences for "Blueberries" in Meridiani Planum Sediments

    Science.gov (United States)

    Coleman, M. L.

    2005-03-01

    Formation of spheroidal concretions on Earth results generally from reactions of organic matter in oxidized sediments. Had organic matter been present in Merididani Planum it could have produced a reduced iron mineral phase later oxidized to hematite.

  9. Study of human prostate spheroids treated with zinc using X-ray microfluorescence

    International Nuclear Information System (INIS)

    Leitao, Roberta G.; Lopes, Ricardo T.; Pereira, Gabriela R.; Santos, Carlos A.N.; Palumbo Junior, Antonio; Nasciutti, Luiz E.; Souza, Pedro A.V.R.; Anjos, Marcelino J.

    2013-01-01

    Spheroids cell culture is a useful technique for tissue engineering or regenerative medicine re-search, pharmacological and toxicological studies, and fundamental studies in cell biology. In this study, we investigated Zn distribution in cell spheroids in benign prostate hyperplasia (BPH) and prostate cancer (DU145) and analyzed the differences in the response to Zinc (0-150 μM) treatment. The measurements were performed in standard geometry of 45 deg incidence, exciting with a white beam and using an optical capillary with 20 μm diameter collimation in the XRF beam line at the Synchrotron Light National Laboratory (Campinas, Brazil). The results showed non-uniform distribution of Zn in all the spheroids analyzed. The differential response to zinc of DU145 and BPH cell spheroids suggests that zinc may have an important role in prostate cancer and BPH diagnosis. (author)

  10. Repair, redistribution and repopulation in V79 spheroids during multifraction irradiation

    International Nuclear Information System (INIS)

    Brown, R.C.; Durand, R.E.

    1994-01-01

    We used cells growing as multicell spheroids to determine whether the initial radiation response would be predictive for multifraction exposures, or whether other factors including repopulation rate should be considered. Potential problems of hypoxia and reoxygenation were avoided by using small spheroids which had not yet developed radiobiologically hypoxic regions. Repair and redistribution dominated the responses in the first two or three exposures, with repopulation playing a minor role. As the fractionation schedule was extended, however, repopulation between fractions largely determined the number of viable cells per spheroid. We conclude that the radiation response of cells from untreated spheroids provides a general indication of net sensitivity, but that repair and redistribution produces considerable variation in radiosensitivity throughout a fractionation protocol. Ultimately, repopulation effects may dominate the multifraction response. (Author)

  11. Direct Measurements of Oxygen Gradients in Spheroid Culture System Using Electron Parametric Resonance Oximetry.

    Directory of Open Access Journals (Sweden)

    Laura M Langan

    Full Text Available Advanced in vitro culture from tissues of different origin includes three-dimensional (3D organoid micro structures that may mimic conditions in vivo. One example of simple 3D culture is spheroids; ball shaped structures typically used as liver and tumour models. Oxygen is critically important in physiological processes, but is difficult to quantify in 3D culture: and the question arises, how small does a spheroid have to be to have minimal micro-environment formation? This question is of particular importance in the growing field of 3D based models for toxicological assessment. Here, we describe a simple non-invasive approach modified for the quantitative measurement and subsequent evaluation of oxygen gradients in spheroids developed from a non-malignant fish cell line (i.e. RTG-2 cells using Electron Paramagnetic Resonance (EPR oximetry. Sonication of the paramagnetic probe Lithium phthalocyanine (LiPc allows for incorporation of probe particulates into spheroid during its formation. Spectra signal strength after incorporation of probe into spheroid indicated that a volume of 20 μl of probe (stock solution: 0.10 mg/mL is sufficient to provide a strong spectra across a range of spheroid sizes. The addition of non-toxic probes (that do not produce or consume oxygen report on oxygen diffusion throughout the spheroid as a function of size. We provide evidence supporting the use of this model over a range of initial cell seeding densities and spheroid sizes with the production of oxygen distribution as a function of these parameters. In our spheroid model, lower cell seeding densities (∼2,500 cells/spheroid and absolute size (118±32 μm allow control of factors such as pre-existing stresses (e.g. ∼ 2% normoxic/hypoxic interface for more accurate measurement of treatment response. The applied methodology provides an elegant, widely applicable approach to directly characterize spheroid (and other organoid cultures in biomedical and

  12. Axisymmetric scattering of an acoustical Bessel beam by a rigid fixed spheroid

    OpenAIRE

    Mitri, F. G.

    2015-01-01

    Based on the partial-wave series expansion (PWSE) method in spherical coordinates, a formal analytical solution for the acoustic scattering of a zeroth-order Bessel acoustic beam centered on a rigid fixed (oblate or prolate) spheroid is provided. The unknown scattering coefficients of the spheroid are determined by solving a system of linear equations derived for the Neumann boundary condition. Numerical results for the modulus of the backscattered pressure (\\theta = \\pi) in the near-field an...

  13. Theory of electrostatic fluid modes in a cold spheroidal non-neutral plasma

    International Nuclear Information System (INIS)

    Dubin, D.H.E.

    1991-01-01

    The normal modes of a magnetized spheroidally shaped pure ion plasma have recently been measured. Here the theory of these modes is presented. Although one might expect that a numerical solution is required (because the plasma dielectric is anisotropic and the plasma is inhomogeneous), the problem is actually separable in an unusual coordinate system. The result is a simple electrostatic fluid dispersion relation for modes in a cloud of any spheroidal shape

  14. Radioresistance of human glioma spheroids and expression of HSP70, p53 and EGFr

    International Nuclear Information System (INIS)

    Fedrigo, Carlos A; Rocha, Adriana B da; Grivicich, Ivana; Schunemann, Daniel P; Chemale, Ivan M; Santos, Daiane dos; Jacovas, Thais; Boschetti, Patryck S; Jotz, Geraldo P; Filho, Aroldo Braga

    2011-01-01

    Radiation therapy is routinely prescribed for high-grade malignant gliomas. However, the efficacy of this therapeutic modality is often limited by the occurrence of radioresistance, reflected as a diminished susceptibility of the irradiated cells to undergo cell death. Thus, cells have evolved an elegant system in response to ionizing radiation induced DNA damage, where p53, Hsp70 and/or EGFr may play an important role in the process. In the present study, we investigated whether the content of p53, Hsp70 and EGFr are associated to glioblastoma (GBM) cell radioresistance. Spheroids from U-87MG and MO59J cell lines as well as spheroids derived from primary culture of tumor tissue of one GBM patient (UGBM1) were irradiated (5, 10 and 20 Gy), their relative radioresistance were established and the p53, Hsp70 and EGFr contents were immunohistochemically determined. Moreover, we investigated whether EGFr-phospho-Akt and EGFr-MEK-ERK pathways can induce GBM radioresistance using inhibitors of activation of ERK (PD098059) and Akt (wortmannin). At 5 Gy irradiation UGBM1 and U-87MG spheroids showed growth inhibition whereas the MO59J spheroid was relatively radioresistant. Overall, no significant changes in p53 and Hsp70 expression were found following 5 Gy irradiation treatment in all spheroids studied. The only difference observed in Hsp70 content was the periphery distribution in MO59J spheroids. However, 5 Gy treatment induced a significant increase on the EGFr levels in MO59J spheroids. Furthermore, treatment with inhibitors of activation of ERK (PD098059) and Akt (wortmannin) leads to radiosensitization of MO59J spheroids. These results indicate that the PI3K-Akt and MEK-ERK pathways triggered by EGFr confer GBM radioresistance

  15. High-throughput image analysis of tumor spheroids: a user-friendly software application to measure the size of spheroids automatically and accurately.

    Science.gov (United States)

    Chen, Wenjin; Wong, Chung; Vosburgh, Evan; Levine, Arnold J; Foran, David J; Xu, Eugenia Y

    2014-07-08

    The increasing number of applications of three-dimensional (3D) tumor spheroids as an in vitro model for drug discovery requires their adaptation to large-scale screening formats in every step of a drug screen, including large-scale image analysis. Currently there is no ready-to-use and free image analysis software to meet this large-scale format. Most existing methods involve manually drawing the length and width of the imaged 3D spheroids, which is a tedious and time-consuming process. This study presents a high-throughput image analysis software application - SpheroidSizer, which measures the major and minor axial length of the imaged 3D tumor spheroids automatically and accurately; calculates the volume of each individual 3D tumor spheroid; then outputs the results in two different forms in spreadsheets for easy manipulations in the subsequent data analysis. The main advantage of this software is its powerful image analysis application that is adapted for large numbers of images. It provides high-throughput computation and quality-control workflow. The estimated time to process 1,000 images is about 15 min on a minimally configured laptop, or around 1 min on a multi-core performance workstation. The graphical user interface (GUI) is also designed for easy quality control, and users can manually override the computer results. The key method used in this software is adapted from the active contour algorithm, also known as Snakes, which is especially suitable for images with uneven illumination and noisy background that often plagues automated imaging processing in high-throughput screens. The complimentary "Manual Initialize" and "Hand Draw" tools provide the flexibility to SpheroidSizer in dealing with various types of spheroids and diverse quality images. This high-throughput image analysis software remarkably reduces labor and speeds up the analysis process. Implementing this software is beneficial for 3D tumor spheroids to become a routine in vitro model

  16. Arbitrary scattering of an acoustical Bessel beam by a rigid spheroid with large aspect-ratio

    Science.gov (United States)

    Gong, Zhixiong; Li, Wei; Mitri, Farid G.; Chai, Yingbin; Zhao, Yao

    2016-11-01

    In this paper, the T-matrix (null-field) method is applied to investigate the acoustic scattering by a large-aspect-ratio rigid spheroid immersed in a non-viscous fluid under the illumination of an unbounded zeroth-order Bessel beam with arbitrary orientation. Based on the proposed method, a MATLAB software package is constructed accordingly, and then verified and validated to compute the acoustic scattering by a rigid oblate or prolate spheroid in the Bessel beam. Several numerical examples are carried out to investigate the novel phenomenon of acoustic scattering by spheroids in Bessel beams with arbitrary incidence, with particular emphasis on the aspect ratio (i.e. the ratio of the polar radius over the equatorial radius of the spheroid), the half-cone angle of Bessel beam, the dimensionless frequency, as well as the angle of incidence. The quasi-periodic oscillations are observed in the plots of the far-field backscattering form function modulus versus the dimensionless frequency, owing to the interference between the specular reflection and the Franz wave circumnavigating the spheroid in the surrounding fluid. Furthermore, the 3D far-field scattering directivity patterns at end-on incidence and 2D polar plots at arbitrary angles of incidence are exhibited, which could provide new insights into the physical mechanisms of Bessel beam scattering by flat or elongated spheroid. This research work may provide an impetus for the application of acoustic Bessel beam in engineering practices.

  17. The effect of spheroidizing by thermal cycling in low concentration Cr-Mo alloy steel

    International Nuclear Information System (INIS)

    Yun, H.S.; Kang, C.Y.

    1979-01-01

    An intensive study was carried out on spheroidizing of pearlite (Sph) and number of spherical carbide in proeutectoid ferrite (No/100) of low concentration Cr-Mo steel with thermal cycling. Physical and mechanical properties of steel containing 0.33 % C with thermal cycling were compared with those of low concentration Cr-Mo steel with thermal cycling. The effect of normal heat treatment and cooling rate on spheroidizing of pearlite and precipitation of fine spherical carbide in the steels were investigated. The results obtained were as follows: 1) Thermal cycling of low concentration Cr-Mo steel promoted the spheroidizing of pearlite compared with that of steel without Cr and Mo to steel had significant effect on spheroidizing of pearlite. 2) Number of fine spherical carbides of low concentration Cr-Mo steel with thermal cycling was over 5 times to that of fine spherical carbides of hypoeutectoid steel with thermal cycling. 3) Spheroidizing of pearlite and number of fine spherical carbide in proeutectoid ferrite of low concentration Cr-Mo steel with increasing thermal cycle and cooling rate. 4) Hardness of steel with thermal cycling was decreased. However, low concentration Cr-Mo steel had little decreasing rate in hardness with increasing thermal cycle on the basis of 100 times in thermal cycle. Therefore, toughness was considered to be increased with increasing spheroidizing of pearlite without changing mechanical properties. (author)

  18. Cancer cell spheroids are a better screen for the photodynamic efficiency of glycosylated photosensitizers.

    Directory of Open Access Journals (Sweden)

    Patrícia M R Pereira

    Full Text Available Photodynamic Therapy (PDT relies on the use of non-toxic photosensitizers that are locally and selectively activated by light to induce cell death or apoptosis through reactive oxygen species generation. The conjugation of porphyrinoids with sugars that target cancer is increasingly viewed as an effective way to increase the selectivity of PDT. To date, in vitro PDT efficacy is mostly screened using two-dimensional monolayer cultures. Compared to monolayer cultures, three-dimensional spheroid cultures have unique spatial distributions of nutrients, metabolites, oxygen and signalling molecules; therefore better mimic in vivo conditions. We obtained 0.05 mm3 spheroids with four different human tumor cell lines (HCT-116, MCF-7, UM-UC-3 and HeLa with appropriate sizes for screening PDT agents. We observed that detachment from monolayer culture and growth as tumor spheroids was accompanied by changes in glucose metabolism, endogenous ROS levels, galectin-1 and glucose transporter GLUT1 protein levels. We compared the phototoxic responses of a porphyrin conjugated with four glucose molecules (PorGlu4 in monolayer and spheroid cultures. The uptake and phototoxicity of PorGlu4 is highly dependent on the monolayer versus spheroid model used and on the different levels of GLUT1 protein expressed by these in vitro platforms. This study demonstrates that HCT-116, MCF-7, UM-UC-3 and HeLa spheroids afford a more rational platform for the screening of new glycosylated-photosensitizers compared to monolayer cultures of these cancer cells.

  19. The triple helix of collagens - an ancient protein structure that enabled animal multicellularity and tissue evolution.

    Science.gov (United States)

    Fidler, Aaron L; Boudko, Sergei P; Rokas, Antonis; Hudson, Billy G

    2018-04-09

    The cellular microenvironment, characterized by an extracellular matrix (ECM), played an essential role in the transition from unicellularity to multicellularity in animals (metazoans), and in the subsequent evolution of diverse animal tissues and organs. A major ECM component are members of the collagen superfamily -comprising 28 types in vertebrates - that exist in diverse supramolecular assemblies ranging from networks to fibrils. Each assembly is characterized by a hallmark feature, a protein structure called a triple helix. A current gap in knowledge is understanding the mechanisms of how the triple helix encodes and utilizes information in building scaffolds on the outside of cells. Type IV collagen, recently revealed as the evolutionarily most ancient member of the collagen superfamily, serves as an archetype for a fresh view of fundamental structural features of a triple helix that underlie the diversity of biological activities of collagens. In this Opinion, we argue that the triple helix is a protein structure of fundamental importance in building the extracellular matrix, which enabled animal multicellularity and tissue evolution. © 2018. Published by The Company of Biologists Ltd.

  20. Noise-induced effects on multicellular biopacemaker spontaneous activity: Differences between weak and strong pacemaker cells

    Science.gov (United States)

    Aghighi, Alireza; Comtois, Philippe

    2017-09-01

    Self-organization of spontaneous activity of a network of active elements is important to the general theory of reaction-diffusion systems as well as for pacemaking activity to initiate beating of the heart. Monolayer cultures of neonatal rat ventricular myocytes, consisting of resting and pacemaker cells, exhibit spontaneous activation of their electrical activity. Similarly, one proposed approach to the development of biopacemakers as an alternative to electronic pacemakers for cardiac therapy is based on heterogeneous cardiac cells with resting and spontaneously beating phenotypes. However, the combined effect of pacemaker characteristics, density, and spatial distribution of the pacemaker cells on spontaneous activity is unknown. Using a simple stochastic pattern formation algorithm, we previously showed a clear nonlinear dependency of spontaneous activity (occurrence and amplitude of spontaneous period) on the spatial patterns of pacemaker cells. In this study, we show that this behavior is dependent on the pacemaker cell characteristics, with weaker pacemaker cells requiring higher density and larger clusters to sustain multicellular activity. These multicellular structures also demonstrated an increased sensitivity to voltage noise that favored spontaneous activity at lower density while increasing temporal variation in the period of activity. This information will help researchers overcome the current limitations of biopacemakers.

  1. Plug-and-Play Multicellular Circuits with Time-Dependent Dynamic Responses.

    Science.gov (United States)

    Urrios, Arturo; Gonzalez-Flo, Eva; Canadell, David; de Nadal, Eulàlia; Macia, Javier; Posas, Francesc

    2018-04-20

    Synthetic biology studies aim to develop cellular devices for biomedical applications. These devices, based on living instead of electronic or electromechanic technology, might provide alternative treatments for a wide range of diseases. However, the feasibility of these devices depends, in many cases, on complex genetic circuits that must fulfill physiological requirements. In this work, we explored the potential of multicellular architectures to act as an alternative to complex circuits for implementation of new devices. As a proof of concept, we developed specific circuits for insulin or glucagon production in response to different glucose levels. Here, we show that fundamental features, such as circuit's affinity or sensitivity, are dependent on the specific configuration of the multicellular consortia, providing a method for tuning these properties without genetic engineering. As an example, we have designed and built circuits with an incoherent feed-forward loop architecture (FFL) that can be easily adjusted to generate single pulse responses. Our results might serve as a blueprint for future development of cellular devices for glycemia regulation in diabetic patients.

  2. Ability of multicellular salt glands in Tamarix species to secrete Na+ and K+ selectively.

    Science.gov (United States)

    Ma, Haiyan; Tian, Changyan; Feng, Gu; Yuan, Junfeng

    2011-03-01

    The present study aimed to determine the mechanism of cation-selective secretion by multicellular salt glands. Using a hydroponic culture system, the secretion and accumulation of Na(+) and K(+) in Tamarix ramosissima and T. laxa under different salt stresses (NaCl, KCl and NaCl+KCl) were studied. Additionally, the effects of salt gland inhibitors (orthovanadate, Ba(2+), ouabain, tetraethylammonium (TEA) and verapamil) on Na(+) and K(+) secretion and accumulation were examined. Treatment with NaCl (at 0-200 mmol L(-1) levels) significantly increased Na(+) secretion, whereas KCl treatment (at 0-200 mmol L(-1) levels) significantly increased K(+) secretion. The ratio of secretion to accumulation of Na(+) was higher than that of K(+). The changes in Na(+) and K(+) secretion differed after adding different ions into the single-salt solutions. Addition of NaCl to the KCl solution (at 100 mmol L(-1) level, respectively) led to a significant decrease in K(+) secretion rate, whereas addition of KCl to the NaCl solution (at 100 mmol L(-1) level, respectively) had little impact on the Na(+) secretion rate. These results indicated that Na+ secretion in Tamarix was highly selective. In addition, Na(+) secretion was significantly inhibited by orthovanadate, ouabain, TEA and verapamil, and K(+) secretion was significantly inhibited by ouabain, TEA and verapamil. The different impacts of orthovanadate on Na(+) and K(+) secretion might be the primary cause for the different Na(+) and K(+) secretion abilities of multicellular salt glands in Tamarix.

  3. Multistage model for the action of cytotoxic T lymphocytes in multicellular conjugates

    International Nuclear Information System (INIS)

    Macken, C.A.; Perelson, A.S.

    1984-01-01

    The authors propose a multistage stochastic model to explain data on the kinetics of target cell lysis by cytotoxic T lymphocytes in multicellular conjugates. A novel feature of this model is that the authors explicitly consider both the lethal hitting stage and the target cell disintegration stage of the cytolytic process. Further, the authors allow for the possibility that target cell disintegration is itself a complex process composed of many events. The comparison of this model with the data of other investigators suggests that cytotoxic T cells deliver lethal hits at random to undamage target cells. Having received a lethal hit, the target cell disintegrates over a variable length of time. The disintegration times of target cells from different conjugates appear to be randomly distributed and to be consistent with a model in which disintegration occurs by at least two major, sequential, rate-limiting events. For conjugates containing one lymphocyte and multiple target cells, the mean rate at which a lethally hit target cell disintegrates is found to be independent of the total number of target cells in the conjugate. This model predicts that in such multicellular conjugates, individual target cells lyse one by one, on average at approximately 30-min intervals, thus agreeing closely with previously reported experimental observations. 35 references, 3 figures, 2 tables

  4. Inter-specific coral chimerism: Genetically distinct multicellular structures associated with tissue loss in Montipora capitata

    Science.gov (United States)

    Work, Thierry M.; Forsman, Zac H.; Szabo, Zoltan; Lewis, Teresa D.; Aeby, Greta S.; Toonen, Robert J.

    2011-01-01

    Montipora white syndrome (MWS) results in tissue-loss that is often lethal to Montipora capitata, a major reef building coral that is abundant and dominant in the Hawai'ian Archipelago. Within some MWS-affected colonies in Kane'ohe Bay, Oahu, Hawai'i, we saw unusual motile multicellular structures within gastrovascular canals (hereafter referred to as invasive gastrovascular multicellular structure-IGMS) that were associated with thinning and fragmentation of the basal body wall. IGMS were in significantly greater densities in coral fragments manifesting tissue-loss compared to paired normal fragments. Mesenterial filaments from these colonies yielded typical M. capitata mitochondrial haplotypes (CO1, CR), while IGMS from the same colony consistently yielded distinct haplotypes previously only found in a different Montipora species (Montipora flabellata). Protein profiles showed consistent differences between paired mesenterial filaments and IGMS from the same colonies as did seven microsatellite loci that also exhibited an excess of alleles per locus inconsistent with a single diploid organism. We hypothesize that IGMS are a parasitic cellular lineage resulting from the chimeric fusion between M. capitata and M. flabellata larvae followed by morphological reabsorption of M. flabellata and subsequent formation of cell-lineage parasites. We term this disease Montiporaiasis. Although intra-specific chimerism is common in colonial animals, this is the first suspected inter-specific example and the first associated with tissue loss.

  5. From cat's eyes to disjoint multicellular natural convection flow in tall tilted cavities

    Energy Technology Data Exchange (ETDEWEB)

    Nicolas, Alfredo, E-mail: anc@xanum.uam.mx [Depto. Matematicas, 3er Piso Ed. AT-Diego Bricio, UAM-I, 09340 Mexico D.F. (Mexico); Baez, Elsa [Depto. Matematicas Aplicadas y Sistemas, UAM-C, 01120 Mexico D.F. (Mexico); Bermudez, Blanca [Facultad de C. de la Computacion, BUAP, 72570 Puebla, Pue. (Mexico)

    2011-07-11

    Numerical results of two-dimensional natural convection problems, in air-filled tall cavities, are reported to study the change of the cat's eyes flow as some parameters vary, the aspect ratio A and the angle of inclination φ of the cavity, with the Rayleigh number Ra mostly fixed; explicitly, the range of the variation is given by 12≤A≤20 and 0{sup o}≤φ≤270{sup o}; about Ra=1.1x10{sup 4}. A novelty contribution of this work is the transition from the cat's eyes changes, as A varies, to a disjoint multicellular flow, as φ varies. These flows may be modeled by the unsteady Boussinesq approximation in stream function and vorticity variables which is solved with a fixed point iterative process applied to the nonlinear elliptic system that results after time discretization. The validation of the results relies on mesh size and time-step independence studies. -- Highlights: → Fixed point iterative method for solving symmetric linear elliptic problems. → Robust method to study effects as aspect ratio and angle of inclination vary. → Interest on the dynamics and evolution of the fluid flow and on heat transfer. → Study of transition from cat's eyes instability to a disjoint multicellular flow.

  6. RssAB signaling coordinates early development of surface multicellularity in Serratia marcescens.

    Directory of Open Access Journals (Sweden)

    Yu-Huan Tsai

    Full Text Available Bacteria can coordinate several multicellular behaviors in response to environmental changes. Among these, swarming and biofilm formation have attracted significant attention for their correlation with bacterial pathogenicity. However, little is known about when and where the signaling occurs to trigger either swarming or biofilm formation. We have previously identified an RssAB two-component system involved in the regulation of swarming motility and biofilm formation in Serratia marcescens. Here we monitored the RssAB signaling status within single cells by tracing the location of the translational fusion protein EGFP-RssB following development of swarming or biofilm formation. RssAB signaling is specifically activated before surface migration in swarming development and during the early stage of biofilm formation. The activation results in the release of RssB from its cognate inner membrane sensor kinase, RssA, to the cytoplasm where the downstream gene promoters are located. Such dynamic localization of RssB requires phosphorylation of this regulator. By revealing the temporal activation of RssAB signaling following development of surface multicellular behavior, our findings contribute to an improved understanding of how bacteria coordinate their lifestyle on a surface.

  7. Hazard identification of exhausts from gasoline-ethanol fuel blends using a multi-cellular human lung model.

    Science.gov (United States)

    Bisig, Christoph; Roth, Michèle; Müller, Loretta; Comte, Pierre; Heeb, Norbert; Mayer, Andreas; Czerwinski, Jan; Petri-Fink, Alke; Rothen-Rutishauser, Barbara

    2016-11-01

    Ethanol can be produced from biomass and as such is renewable, unlike petroleum-based fuel. Almost all gasoline cars can drive with fuel containing 10% ethanol (E10), flex-fuel cars can even use 85% ethanol (E85). Brazil and the USA already include 10-27% ethanol in their standard fuel by law. Most health effect studies on car emissions are however performed with diesel exhausts, and only few data exists for other fuels. In this work we investigated possible toxic effects of exhaust aerosols from ethanol-gasoline blends using a multi-cellular model of the human lung. A flex-fuel passenger car was driven on a chassis dynamometer and fueled with E10, E85, or pure gasoline (E0). Exhausts obtained from a steady state cycle were directly applied for 6h at a dilution of 1:10 onto a multi-cellular human lung model mimicking the bronchial compartment composed of human bronchial cells (16HBE14o-), supplemented with human monocyte-derived dendritic cells and monocyte-derived macrophages, cultured at the air-liquid interface. Biological endpoints were assessed after 6h post incubation and included cytotoxicity, pro-inflammation, oxidative stress, and DNA damage. Filtered air was applied to control cells in parallel to the different exhausts; for comparison an exposure to diesel exhaust was also included in the study. No differences were measured for the volatile compounds, i.e. CO, NO x , and T.HC for the different ethanol supplemented exhausts. Average particle number were 6×10 2 #/cm 3 (E0), 1×10 5 #/cm 3 (E10), 3×10 3 #/cm 3 (E85), and 2.8×10 6 #/cm 3 (diesel). In ethanol-gasoline exposure conditions no cytotoxicity and no morphological changes were observed in the lung cell cultures, in addition no oxidative stress - as analyzed with the glutathione assay - was measured. Gene expression analysis also shows no induction in any of the tested genes, including mRNA levels of genes related to oxidative stress and pro-inflammation, as well as indoleamine 2,3-dioxygenase 1

  8. Hepatocyte spheroids as a competent in vitro system for drug biotransformation studies: nevirapine as a bioactivation case study.

    Science.gov (United States)

    Pinheiro, Pedro F; Pereira, Sofia A; Harjivan, Shrika G; Martins, Inês L; Marinho, Aline T; Cipriano, Madalena; Jacob, Cristina C; Oliveira, Nuno G; Castro, Matilde F; Marques, M Matilde; Antunes, Alexandra M M; Miranda, Joana P

    2017-03-01

    The development of metabolically competent in vitro models is of utmost importance for predicting adverse drug reactions, thereby preventing attrition-related economical and clinical burdens. Using the antiretroviral drug nevirapine (NVP) as a model, this work aimed to validate rat hepatocyte 3D spheroid cultures as competent in vitro systems to assess drug metabolism and bioactivation. Hepatocyte spheroids were cultured for 12 days in a stirred tank system (3D cultures) and exposed to equimolar dosages of NVP and its two major Phase I metabolites, 12-OH-NVP and 2-OH-NVP. Phase I NVP metabolites were detected in the 3D cultures during the whole culture time in the same relative proportions reported in in vivo studies. Moreover, the modulation of SULT1A1 activity by NVP and 2-OH-NVP was observed for the first time, pointing their synergistic effect as a key factor in the formation of the toxic metabolite (12-sulfoxy-NVP). Covalent adducts formed by reactive NVP metabolites with N-acetyl-L-cysteine and bovine serum albumin were also detected by high-resolution mass spectrometry, providing new evidence on the relative role of the reactive NVP metabolites, 12-sulfoxy-NVP, and NVP quinone methide, in toxicity versus excretion pathways. In conclusion, these results demonstrate the validity of the 3D culture system to evaluate drug bioactivation, enabling the identification of potential biomarkers of bioactivation/toxicity, and providing new evidence to the mechanisms underlying NVP-induced toxic events. This model, integrated with the analytical strategies described herein, is of anticipated usefulness to the pharmaceutical industry, as an upstream methodology for flagging drug safety alerts in early stages of drug development.

  9. Universal Dark Halo Scaling Relation for the Dwarf Spheroidal Satellites

    Science.gov (United States)

    Hayashi, Kohei; Ishiyama, Tomoaki; Ogiya, Go; Chiba, Masashi; Inoue, Shigeki; Mori, Masao

    2017-07-01

    Motivated by a recently found interesting property of the dark halo surface density within a radius, {r}\\max , giving the maximum circular velocity, {V}\\max , we investigate it for dark halos of the Milky Way’s and Andromeda’s dwarf satellites based on cosmological simulations. We select and analyze the simulated subhalos associated with Milky-Way-sized dark halos and find that the values of their surface densities, {{{Σ }}}{V\\max }, are in good agreement with those for the observed dwarf spheroidal satellites even without employing any fitting procedures. Moreover, all subhalos on the small scales of dwarf satellites are expected to obey the universal relation, irrespective of differences in their orbital evolutions, host halo properties, and observed redshifts. Therefore, we find that the universal scaling relation for dark halos on dwarf galaxy mass scales surely exists and provides us with important clues for understanding fundamental properties of dark halos. We also investigate orbital and dynamical evolutions of subhalos to understand the origin of this universal dark halo relation and find that most subhalos evolve generally along the {r}\\max \\propto {V}\\max sequence, even though these subhalos have undergone different histories of mass assembly and tidal stripping. This sequence, therefore, should be the key feature for understanding the nature of the universality of {{{Σ }}}{V\\max }.

  10. Three-dimensional alginate spheroid culture system of murine osteosarcoma.

    Science.gov (United States)

    Akeda, Koji; Nishimura, Akinobu; Satonaka, Haruhiko; Shintani, Ken; Kusuzaki, Katsuyuki; Matsumine, Akihiko; Kasai, Yuichi; Masuda, Koichi; Uchida, Atsumasa

    2009-11-01

    Osteosarcoma (OS) is the most common primary malignant tumor of the bone and often forms pulmonary metastases, which are the most important prognostic factor. For further elucidation of the mechanism underlying the progression and metastasis of human OS, a culture system mimicking the microenvironment of the tumor in vivo is needed. We report a novel three-dimensional (3D) alginate spheroid culture system of murine osteosarcoma. Two different metastatic clones, the parental Dunn and its derivative line LM8, which has a higher metastatic potential to the lungs, were encapsulated in alginate beads to develop the 3D culture system. The beads containing murine OS cells were also transplanted into mice to determine their metastatic potential in vivo. In this culture system, murine OS cells encapsulated in alginate beads were able to grow in a 3D structure with cells detaching from the alginate environment. The number of detaching cells was higher in the LM8 cell line than the Dunn cell line. In the in vivo alginate bead transplantation model, the rate of pulmonary metastasis was higher with LM8 cells compared with that of Dunn cells. The cell characteristics and kinetics in this culture system closely reflect the original malignant potential of the cells in vivo.

  11. ON THE r -PROCESS ENRICHMENT OF DWARF SPHEROIDAL GALAXIES

    Energy Technology Data Exchange (ETDEWEB)

    Bramante, Joseph [Department of Physics, 225 Nieuwland Science Hall, University of Notre Dame, Notre Dame, IN 46556 (United States); Linden, Tim [Center for Cosmology and AstroParticle Physics (CCAPP) and Department of Physics The Ohio State University, Columbus OH, 43210 (United States)

    2016-07-20

    Recent observations of Reticulum II have uncovered an overabundance of r -process elements compared to similar ultra-faint dwarf spheroidal galaxies (UFDs). Because the metallicity and star formation history of Reticulum II appear consistent with all known UFDs, the high r -process abundance of Reticulum II suggests enrichment through a single, rare event, such as a double neutron star (NS) merger. However, we note that this scenario is extremely unlikely, as binary stellar evolution models require significant supernova natal kicks to produce NS–NS or NS–black hole (BH) mergers, and these kicks would efficiently remove compact binary systems from the weak gravitational potentials of UFDs. We examine alternative mechanisms for the production of r -process elements in UFDs, including a novel mechanism wherein NSs in regions of high dark matter (DM) density implode after accumulating a BH-forming mass of DM. We find that r -process proto-material ejection by tidal forces, when a single NS implodes into a BH, can occur at a rate matching the r -process abundance of both Reticulum II and the Milky Way. Remarkably, DM models which collapse a single NS in observed UFDs also solve the missing pulsar problem in the Milky Way Galactic Center. We propose tests specific to DM r -process production which may uncover or rule out this model.

  12. The Variable Stars of the Draco Dwarf Spheroidal Galaxy: Revisited

    Science.gov (United States)

    Kinemuchi, K.; Harris, H. C.; Smith, Horace A.; Silbermann, N. A.; Snyder, L. A.; La Cluyzé, A. P.; Clark, C. L.

    2008-11-01

    We present a CCD survey of variable stars in the Draco dwarf spheroidal galaxy. This survey, which has the largest areal coverage since the original variable star survey by Baade & Swope, includes photometry for 270 RR Lyrae (RRL) stars, 9 anomalous Cepheids (ACs), 2 eclipsing binaries, and 12 slow, irregular red variables, as well as 30 background QSOs. Twenty-six probable double-mode RRL stars were identified. Observed parameters, including mean V and I magnitudes, V amplitudes, and periods, have been derived. Photometric metallicities of the ab-type RRL stars were calculated according to the method of Jurcsik & Kovacs, yielding a mean metallicity of lang[Fe/H]rang = -2.19 ± 0.03. The well-known Oosterhoff intermediate nature of the RRL stars in Draco is reconfirmed, although the double-mode RRL stars, with one exception, have properties similar to those found in Oosterhoff type II globular clusters. The period-luminosity relation of the ACs is rediscussed with the addition of the new Draco ACs.

  13. A Kinematic Study of the Andromeda Dwarf Spheroidal System

    Science.gov (United States)

    Collins, Michelle L. M.; Chapman, Scott C.; Rich, R. Michael; Ibata, Rodrigo A.; Martin, Nicolas F.; Irwin, Michael J.; Bate, Nicholas F.; Lewis, Geraint F.; Peñarrubia, Jorge; Arimoto, Nobuo; Casey, Caitlin M.; Ferguson, Annette M. N.; Koch, Andreas; McConnachie, Alan W.; Tanvir, Nial

    2013-05-01

    We present a homogeneous kinematic analysis of red giant branch stars within 18 of the 28 Andromeda dwarf spheroidal (dSph) galaxies, obtained using the Keck I/LRIS and Keck II/DEIMOS spectrographs. Based on their g - i colors (taken with the CFHT/MegaCam imager), physical positions on the sky, and radial velocities, we assign probabilities of dSph membership to each observed star. Using this information, the velocity dispersions, central masses, and central densities of the dark matter halos are calculated for these objects, and compared with the properties of the Milky Way dSph population. We also measure the average metallicity ([Fe/H]) from the co-added spectra of member stars for each M31 dSph and find that they are consistent with the trend of decreasing [Fe/H] with luminosity observed in the Milky Way population. We find that three of our studied M31 dSphs appear as significant outliers in terms of their central velocity dispersion, And XIX, XXI, and XXV, all of which have large half-light radii (gsim 700 pc) and low velocity dispersions (σ v ratio within its half-light radius of just [M/L]_half=10.3^{+7.0}_{-6.7}, making it consistent with a simple stellar system with no appreciable dark matter component within its 1σ uncertainties. We suggest that the structure of the dark matter halos of these outliers have been significantly altered by tides.

  14. A Chemical Evolution Model for the Fornax Dwarf Spheroidal Galaxy

    Directory of Open Access Journals (Sweden)

    Yuan Zhen

    2016-01-01

    Full Text Available Fornax is the brightest Milky Way (MW dwarf spheroidal galaxy and its star formation history (SFH has been derived from observations. We estimate the time evolution of its gas mass and net inflow and outflow rates from the SFH usinga simple star formation law that relates the star formation rate to the gas mass. We present a chemical evolution model on a 2D mass grid with supernovae (SNe as sources of metal enrichment. We find that a key parameter controlling the enrichment is the mass Mx of the gas to mix with the ejecta from each SN. The choice of Mx depends on the evolution of SN remnants and on the global gas dynamics. It differs between the two types of SNe involved and between the periods before and after Fornax became an MW satellite at time t = tsat. Our results indicate that due to the global gas outflow at t > tsat, part of the ejecta from each SN may directly escape from Fornax. Sample results from our model are presented and compared with data.

  15. Experimental evidence for several spheroid growth mechanisms in the liquid-phase sintered tungsten-base composites

    International Nuclear Information System (INIS)

    Zukas, E.G.; Rogers, P.S.Z.; Rogers, R.S.

    1976-01-01

    The generally accepted mechanism for spheroid growth during sintering of tungsten-base composites in the presence of a liquid phase is the dissolution of the small spheroids with simultaneous precipitation of tungsten from the molten matrix onto the larger spheroids, the process being driven by the difference in surface energy between the larger and smaller spheroids. From theoretical considerations, the slope of the straight line of log diameter versus log time should be 1 / 3 for this process. The experimental evidence for the dissolution and reprecipitation mechanism is meager, being based primarily on the spheroid growth rate during the latter stages of liquid-phase sintering. Experimental evidence is presented that shows spheroid growth taking place in systems where the tungsten and the matrix are mutually insoluble thereby making dissolution and reprecipitation impossible. Furthermore, the results from these studies and others using the usual matrix compositions indicate that spheroid growth takes place predominantly by the combination or coalescence of two or more spheroids. Deposition of tungsten from the molten matrix also occurs, although not necessarily on spheroid surfaces which have the lowest surface energy. Thus, many mechanisms, each depending on temperature and other variables, operate simultaneously. A satisfactory theoretical treatment must include them all

  16. Retrieval of spheroid particle size distribution from spectral extinction data in the independent mode using PCA approach

    International Nuclear Information System (INIS)

    Tang, Hong; Lin, Jian-Zhong

    2013-01-01

    An improved anomalous diffraction approximation (ADA) method is presented for calculating the extinction efficiency of spheroids firstly. In this approach, the extinction efficiency of spheroid particles can be calculated with good accuracy and high efficiency in a wider size range by combining the Latimer method and the ADA theory, and this method can present a more general expression for calculating the extinction efficiency of spheroid particles with various complex refractive indices and aspect ratios. Meanwhile, the visible spectral extinction with varied spheroid particle size distributions and complex refractive indices is surveyed. Furthermore, a selection principle about the spectral extinction data is developed based on PCA (principle component analysis) of first derivative spectral extinction. By calculating the contribution rate of first derivative spectral extinction, the spectral extinction with more significant features can be selected as the input data, and those with less features is removed from the inversion data. In addition, we propose an improved Tikhonov iteration method to retrieve the spheroid particle size distributions in the independent mode. Simulation experiments indicate that the spheroid particle size distributions obtained with the proposed method coincide fairly well with the given distributions, and this inversion method provides a simple, reliable and efficient method to retrieve the spheroid particle size distributions from the spectral extinction data. -- Highlights: ► Improved ADA is presented for calculating the extinction efficiency of spheroids. ► Selection principle about spectral extinction data is developed based on PCA. ► Improved Tikhonov iteration method is proposed to retrieve the spheroid PSD.

  17. Acoustic scattering of a Bessel vortex beam by a rigid fixed spheroid

    Science.gov (United States)

    Mitri, F. G.

    2015-12-01

    Partial-wave series representation of the acoustic scattering field of high-order Bessel vortex beams by rigid oblate and prolate spheroids using the modal matching method is developed. The method, which is applicable to slightly elongated objects at low-to-moderate frequencies, requires solving a system of linear equations which depends on the partial-wave index n and the order of the Bessel vortex beam m using truncated partial-wave series expansions (PWSEs), and satisfying the Neumann boundary condition for a rigid immovable surface in the least-squares sense. This original semi-analytical approach developed for Bessel vortex beams is demonstrated for finite oblate and prolate spheroids, where the mathematical functions describing the spheroidal geometry are written in a form involving single angular (polar) integrals that are numerically computed. The transverse (θ = π / 2) and 3D scattering directivity patterns are evaluated in the far-field for both prolate and oblate spheroids, with particular emphasis on the aspect ratio (i.e., the ratio of the major axis over the minor axis of the spheroid) not exceeding 3:1, the half-cone angle β and order m of the Bessel vortex beam, as well as the dimensionless size parameter kr0. Periodic oscillations in the magnitude plots of the far-field scattering form function are observed, which result from the interference of the reflected waves with the circumferential (Franz') waves circumnavigating the surface of the spheroid in the surrounding fluid. Moreover, the 3D directivity patterns illustrate the far-field scattering from the spheroid, that vanishes in the forward (θ = 0) and backward (θ = π) directions. Particular applications in underwater acoustics and scattering, acoustic levitation and the detection of submerged elongated objects using Bessel vortex waves to name a few, would benefit from the results of the present investigation.

  18. Combined exposure of diesel exhaust particles and respirable Soufrière Hills volcanic ash causes a (pro-)inflammatory response in an in vitro multicellular epithelial tissue barrier model

    Science.gov (United States)

    Tomašek, Ines; Horwell, Claire J.; Damby, David; Barošová, Hana; Geers, Christoph; Petri-Fink, Alke; Rothen-Rutishauser, Barbara; Clift, Martin J. D.

    2016-01-01

    BackgroundThere are justifiable health concerns regarding the potential adverse effects associated with human exposure to volcanic ash (VA) particles, especially when considering communities living in urban areas already exposed to heightened air pollution. The aim of this study was, therefore, to gain an imperative, first understanding of the biological impacts of respirable VA when exposed concomitantly with diesel particles.MethodsA sophisticated in vitro 3D triple cell co-culture model of the human alveolar epithelial tissue barrier was exposed to either a single or repeated dose of dry respirable VA (deposited dose of 0.26 ± 0.09 or 0.89 ± 0.29 μg/cm2, respectively) from Soufrière Hills volcano, Montserrat for a period of 24 h at the air-liquid interface (ALI). Subsequently, co-cultures were exposed to co-exposures of single or repeated VA and diesel exhaust particles (DEP; NIST SRM 2975; 0.02 mg/mL), a model urban pollutant, at the pseudo-ALI. The biological impact of each individual particle type was also analysed under these precise scenarios. The cytotoxic (LDH release), oxidative stress (depletion of intracellular GSH) and (pro-)inflammatory (TNF-α, IL-8 and IL-1β) responses were assessed after the particulate exposures. The impact of VA exposure upon cell morphology, as well as its interaction with the multicellular model, was visualised via confocal laser scanning microscopy (LSM) and scanning electron microscopy (SEM), respectively.ResultsThe combination of respirable VA and DEP, in all scenarios, incited an heightened release of TNF-α and IL-8 as well as significant increases in IL-1β, when applied at sub-lethal doses to the co-culture compared to VA exposure alone. Notably, the augmented (pro-)inflammatory responses observed were not mediated by oxidative stress. LSM supported the quantitative assessment of cytotoxicity, with no changes in cell morphology within the barrier model evident. A direct interaction of the VA with all

  19. Implementation of Complex Biological Logic Circuits Using Spatially Distributed Multicellular Consortia

    Science.gov (United States)

    Urrios, Arturo; de Nadal, Eulàlia; Solé, Ricard; Posas, Francesc

    2016-01-01

    Engineered synthetic biological devices have been designed to perform a variety of functions from sensing molecules and bioremediation to energy production and biomedicine. Notwithstanding, a major limitation of in vivo circuit implementation is the constraint associated to the use of standard methodologies for circuit design. Thus, future success of these devices depends on obtaining circuits with scalable complexity and reusable parts. Here we show how to build complex computational devices using multicellular consortia and space as key computational elements. This spatial modular design grants scalability since its general architecture is independent of the circuit’s complexity, minimizes wiring requirements and allows component reusability with minimal genetic engineering. The potential use of this approach is demonstrated by implementation of complex logical functions with up to six inputs, thus demonstrating the scalability and flexibility of this method. The potential implications of our results are outlined. PMID:26829588

  20. Colloquium: Modeling the dynamics of multicellular systems: Application to tissue engineering

    Science.gov (United States)

    Kosztin, Ioan; Vunjak-Novakovic, Gordana; Forgacs, Gabor

    2012-10-01

    Tissue engineering is a rapidly evolving discipline that aims at building functional tissues to improve or replace damaged ones. To be successful in such an endeavor, ideally, the engineering of tissues should be based on the principles of developmental biology. Recent progress in developmental biology suggests that the formation of tissues from the composing cells is often guided by physical laws. Here a comprehensive computational-theoretical formalism is presented that is based on experimental input and incorporates biomechanical principles of developmental biology. The formalism is described and it is shown that it correctly reproduces and predicts the quantitative characteristics of the fundamental early developmental process of tissue fusion. Based on this finding, the formalism is then used toward the optimization of the fabrication of tubular multicellular constructs, such as a vascular graft, by bioprinting, a novel tissue engineering technology.

  1. Small regulatory RNAs control the multi-cellular adhesive lifestyle of Escherichia coli

    DEFF Research Database (Denmark)

    Jørgensen, Mikkel Girke; Nielsen, Jesper Sejrup; Boysen, Anders

    2012-01-01

    Small regulatory RNA molecules have recently been recognized as important regulatory elements of developmental processes in both eukaryotes and bacteria. We here describe a striking example in Escherichia coli that can switch between a single-cell motile lifestyle and a multi-cellular, sessile....... Our demonstration that basal expression of each of the three RNA species is sufficient to downregulate CsgD synthesis and prevent curli formation indicates that all play a prominent role in the curli regulatory network. Our findings provide the first clue as to how the Rcs signalling pathway...... negatively regulates curli synthesis and increase the number of small regulatory RNAs that act directly on the csgD mRNA to five....

  2. Principles of cooperation across systems: from human sharing to multicellularity and cancer.

    Science.gov (United States)

    Aktipis, Athena

    2016-01-01

    From cells to societies, several general principles arise again and again that facilitate cooperation and suppress conflict. In this study, I describe three general principles of cooperation and how they operate across systems including human sharing, cooperation in animal and insect societies and the massively large-scale cooperation that occurs in our multicellular bodies. The first principle is that of Walk Away: that cooperation is enhanced when individuals can leave uncooperative partners. The second principle is that resource sharing is often based on the need of the recipient (i.e., need-based transfers) rather than on strict account-keeping. And the last principle is that effective scaling up of cooperation requires increasingly sophisticated and costly cheater suppression mechanisms. By comparing how these principles operate across systems, we can better understand the constraints on cooperation. This can facilitate the discovery of novel ways to enhance cooperation and suppress cheating in its many forms, from social exploitation to cancer.

  3. Structure and Formation of Elliptical and Spheroidal Galaxies

    Science.gov (United States)

    Kormendy, John; Fisher, David B.; Cornell, Mark E.; Bender, Ralf

    2009-05-01

    dichotomy between elliptical and spheroidal galaxies. Their properties are consistent with our understanding of their different formation processes: mergers for ellipticals and conversion of late-type galaxies into spheroidals by environmental effects and by energy feedback from supernovae. In an appendix, we develop machinery to get realistic error estimates for Sérsic parameters even when they are strongly coupled. And we discuss photometric dynamic ranges necessary to get robust results from Sérsic fits. Based in part on observations obtained with the Hobby-Eberly Telescope (HET), which is a joint project of the University of Texas at Austin, the Pennsylvania State University, Stanford University, Ludwig-Maximilians-Universität München, and Georg-August-Universität Göttingen.

  4. A Duo of Potassium-Responsive Histidine Kinases Govern the Multicellular Destiny of Bacillus subtilis

    Science.gov (United States)

    de Oña, Paula; Kunert, Maritta; Leñini, Cecilia; Gallegos-Monterrosa, Ramses; Mhatre, Eisha; Vileta, Darío; Hölscher, Theresa; Kuipers, Oscar P.

    2015-01-01

    ABSTRACT Multicellular biofilm formation and surface motility are bacterial behaviors considered mutually exclusive. However, the basic decision to move over or stay attached to a surface is poorly understood. Here, we discover that in Bacillus subtilis, the key root biofilm-controlling transcription factor Spo0A~Pi (phosphorylated Spo0A) governs the flagellum-independent mechanism of social sliding motility. A Spo0A-deficient strain was totally unable to slide and colonize plant roots, evidencing the important role that sliding might play in natural settings. Microarray experiments plus subsequent genetic characterization showed that the machineries of sliding and biofilm formation share the same main components (i.e., surfactin, the hydrophobin BslA, exopolysaccharide, and de novo-formed fatty acids). Sliding proficiency was transduced by the Spo0A-phosphorelay histidine kinases KinB and KinC. We discovered that potassium, a previously known inhibitor of KinC-dependent biofilm formation, is the specific sliding-activating signal through a thus-far-unnoticed cytosolic domain of KinB, which resembles the selectivity filter sequence of potassium channels. The differential expression of the Spo0A~Pi reporter abrB gene and the different levels of the constitutively active form of Spo0A, Sad67, in Δspo0A cells grown in optimized media that simultaneously stimulate motile and sessile behaviors uncover the spatiotemporal response of KinB and KinC to potassium and the gradual increase in Spo0A~Pi that orchestrates the sequential activation of sliding, followed by sessile biofilm formation and finally sporulation in the same population. Overall, these results provide insights into how multicellular behaviors formerly believed to be antagonistic are coordinately activated in benefit of the bacterium and its interaction with the host. PMID:26152584

  5. Axisymmetric scattering of an acoustical Bessel beam by a rigid fixed spheroid.

    Science.gov (United States)

    Mitri, Farid G

    2015-10-01

    Based on the partial-wave series expansion (PWSE) method in spherical coordinates, a formal analytical solution for the acoustic scattering of a zeroth-order Bessel acoustic beam centered on a rigid fixed (oblate or prolate) spheroid is provided. The unknown scattering coefficients of the spheroid are determined by solving a system of linear equations derived for the Neumann boundary condition. Numerical results for the modulus of the backscattered pressure (θ = π) in the near field and the backscattering form function in the far field for both prolate and oblate spheroids are presented and discussed, with particular emphasis on the aspect ratio (i.e., the ratio of the major axis over the minor axis of the spheroid), the half-cone angle of the Bessel beam, and the dimensionless frequency. The plots display periodic oscillations (versus the dimensionless frequency) because of the interference of specularly reflected waves in the backscattering direction with circumferential Franz' waves circumnavigating the surface of the spheroid in the surrounding fluid. Moreover, the 3-D directivity patterns illustrate the near- and far-field axisymmetric scattering. Investigations in underwater acoustics, particle levitation, scattering, and the detection of submerged elongated objects and other related applications utilizing Bessel waves would benefit from the results of the present study.

  6. Porous Nb-Ti based alloy produced from plasma spheroidized powder

    Directory of Open Access Journals (Sweden)

    Qijun Li

    Full Text Available Spherical Nb-Ti based alloy powder was prepared by the combination of plasma spheroidization and mechanical alloying. Phase constituents, microstructure and surface state of the powder, and pore characteristics of the resulting porous alloy were investigated. The results show that the undissolved W and V in the mechanically alloyed powder is fully alloyed after spheroidization, and single β phase is achieved. Particle size of the spheroidized powder is in the range of 20–110 μm. With the decrease of particle size, a transformation from typical dendrite solidification structure to fine cell microstructure occurs. The surface of the spheroidized powder is coated by a layer of oxides consisting mainly of TiO2 and Nb2O5. Probabilities of sinter-neck formation and particle coalescence increases with increasing sintering temperature. Porous skeleton with relatively homogeneous pore distribution and open pore channel is formed after vacuum sintering at 1700 °C, and the porosity is 32%. The sintering kinetic analysis indicates that grain boundary diffusion is the primary mass transport mechanism during sintering process. Keywords: Powder metallurgy, Nb-Ti based alloy, Porous material, Mechanical alloying, Plasma spheroidizing, Solidification microstructure

  7. Elemental mapping by synchrotron radiation X-Ray microfluorescence in cellular spheroid of prostate tumor cells

    Energy Technology Data Exchange (ETDEWEB)

    Leitao, R.G.; Anjos, M.J.; Lopes, R.T., E-mail: roberta@lin.ufrj.br [Universidade Federal do Rio de Janeiro (UFRJ), RJ (Brazil). Lab. de Instrumentacao Nuclear; Santos, C.A.N. [Instituto Nacional de Metrologia, Qualidade e Tecnologia (INMETRO), Duque de Caxias, RJ (Brazil). Lab. de Biotecnologia; Palumbo Junior, A.; Souza, P.A.V.R.; Nasciutti, L.E. [Universidade Federal do Rio de Janeiro (UFRJ), RJ (Brazil). Instituto de Ciencias Biomedicas; Pereira, G.R. [Universidade Federal do Rio de Janeiro (UFRJ), RJ (Brazil). Lab. de Ensaios Nao Destrutivos, Corrosao e Soldagem

    2013-08-15

    Prostate cancer is the sixth most common type of cancer and the third most common in males in Western industrialized countries. Cellular spheroid serves as excellent physiologic tumor models as they mimic avascular tumors and micrometastases. Trace elements play a significant role in biological processes. They are capable of affecting human health by competing with essential elements for available binding sites and by the activation or inhibition of reactions between metabolic enzymes. It is well known that zinc levels in the peripheral zone of dorsal and lateral lobes of the prostate are almost 10 times higher than in other soft tissues. Prostate tumor cells were isolated of the prostate tissue samples that were collected from patients submitted to surgery. The measurements were performed in XRF beam line at the Synchrotron Light National Laboratory (LNLS) in Campinas, Brazil. The results showed that all elements were heterogeneously distributed in different areas of the spheroids analyzed. P, S and Cl showed similar elemental distribution in all the samples analyzed while K, Ca, Fe, and Cu showed different elemental distribution. In all spheroids analyzed, Zn presented more intense distributions in the central region of the spheroid. The relationship between the function of Zn in the secretory epithelial cells and the carcinogenic process suggests that more studies on elemental mapping in spheroids are necessary. (author)

  8. Cell invasion in the spheroid sprouting assay: a spatial organisation analysis adaptable to cell behaviour.

    Directory of Open Access Journals (Sweden)

    Silvia Blacher

    Full Text Available The endothelial cell spheroid assay provides a suitable in vitro model to study (lymph angiogenesis and test pro- and anti-(lymph angiogenic factors or drugs. Usually, the extent of cell invasion, observed through optical microscopy, is measured. The present study proposes the spatial distribution of migrated cells as a new descriptor of the (lymph angiogenic response. The utility of this novel method rests with its capacity to locally characterise spheroid structure, allowing not only the investigation of single and collective cell invasion but also the evolution of the spheroid core itself. Moreover, the proposed method can be applied to 2D-projected spheroid images obtained by optical microscopy, as well as to 3D images acquired by confocal microscopy. To validate the proposed methodology, endothelial cell invasion was evaluated under different experimental conditions. The results were compared with widely used global parameters. The comparison shows that our method prevents local spheroid modifications from being overlooked and leading to the possible misinterpretation of results.

  9. Elemental mapping by synchrotron radiation X-Ray microfluorescence in cellular spheroid of prostate tumor cells

    International Nuclear Information System (INIS)

    Leitao, R.G.; Anjos, M.J.; Lopes, R.T.; Santos, C.A.N.; Palumbo Junior, A.; Souza, P.A.V.R.; Nasciutti, L.E.; Pereira, G.R.

    2013-01-01

    Prostate cancer is the sixth most common type of cancer and the third most common in males in Western industrialized countries. Cellular spheroid serves as excellent physiologic tumor models as they mimic avascular tumors and micrometastases. Trace elements play a significant role in biological processes. They are capable of affecting human health by competing with essential elements for available binding sites and by the activation or inhibition of reactions between metabolic enzymes. It is well known that zinc levels in the peripheral zone of dorsal and lateral lobes of the prostate are almost 10 times higher than in other soft tissues. Prostate tumor cells were isolated of the prostate tissue samples that were collected from patients submitted to surgery. The measurements were performed in XRF beam line at the Synchrotron Light National Laboratory (LNLS) in Campinas, Brazil. The results showed that all elements were heterogeneously distributed in different areas of the spheroids analyzed. P, S and Cl showed similar elemental distribution in all the samples analyzed while K, Ca, Fe, and Cu showed different elemental distribution. In all spheroids analyzed, Zn presented more intense distributions in the central region of the spheroid. The relationship between the function of Zn in the secretory epithelial cells and the carcinogenic process suggests that more studies on elemental mapping in spheroids are necessary. (author)

  10. Influence of reaction chamber shape on cast-iron spheroidization process in-mold

    Directory of Open Access Journals (Sweden)

    S. Pietrowski

    2010-01-01

    Full Text Available This paper presents a results concerning the influence of reaction chamber shape on cast – iron spheroidization process in form. The volume of the tested reaction chambers was about 118000mm3. Reaction chambers in the shape of: rectangular, cylinder and spherical cap were examined. It has been shown that the best graphite spheroidizing process was provided by spherical cap chamber shape. The reaction of cast – iron with magnesium in reaction chamber depends on the flow of cast – iron in the chamber. In rectangular and cylinder shape chambers proceed the impact of diphase stream on flat bottom wall. It causes the creation on its surface film, called: cast – iron “film”, where single grains of magnesium master alloy exist. The largest part of master alloy is drifted by liquid cast – iron to the top and only there graphite spheroidization process proceed. In the spherical cap shape reaction chamber, as a result of rotation movement of liquid cast – iron throughout its volume, graphite spheroidization process proceed. Apart from the reaction chamber shape, applying of mixing chamber ensure full cast – iron spheroidization process.

  11. Application of Mie theory to assess structure of spheroidal scattering in backscattering geometries.

    Science.gov (United States)

    Chalut, Kevin J; Giacomelli, Michael G; Wax, Adam

    2008-08-01

    Inverse light scattering analysis seeks to associate measured scattering properties with the most probable theoretical scattering distribution. Although Mie theory is a spherical scattering model, it has been used successfully for discerning the geometry of spheroidal scatterers. The goal of this study was an in-depth evaluation of the consequences of analyzing the structure of spheroidal geometries, which are relevant to cell and tissue studies in biology, by employing Mie-theory-based inverse light scattering analysis. As a basis for this study, the scattering from spheroidal geometries was modeled using T-matrix theory and used as test data. In a previous study, we used this technique to investigate the case of spheroidal scatterers aligned with the optical axis. In the present study, we look at a broader scope which includes the effects of aspect ratio, orientation, refractive index, and incident light polarization. Over this wide range of parameters, our results indicate that this method provides a good estimate of spheroidal structure.

  12. Engineered Breast Cancer Cell Spheroids Reproduce Biologic Properties of Solid Tumors.

    Science.gov (United States)

    Ham, Stephanie L; Joshi, Ramila; Luker, Gary D; Tavana, Hossein

    2016-11-01

    Solid tumors develop as 3D tissue constructs. As tumors grow larger, spatial gradients of nutrients and oxygen and inadequate diffusive supply to cells distant from vasculature develops. Hypoxia initiates signaling and transcriptional alterations to promote survival of cancer cells and generation of cancer stem cells (CSCs) that have self-renewal and tumor-initiation capabilities. Both hypoxia and CSCs are associated with resistance to therapies and tumor relapse. This study demonstrates that 3D cancer cell models, known as tumor spheroids, generated with a polymeric aqueous two-phase system (ATPS) technology capture these important biological processes. Similar to solid tumors, spheroids of triple negative breast cancer cells deposit major extracellular matrix proteins. The molecular analysis establishes presence of hypoxic cells in the core region and expression of CSC gene and protein markers including CD24, CD133, and Nanog. Importantly, these spheroids resist treatment with chemotherapy drugs. A combination treatment approach using a hypoxia-activated prodrug, TH-302, and a chemotherapy drug, doxorubicin, successfully targets drug resistant spheroids. This study demonstrates that ATPS spheroids recapitulate important biological and functional properties of solid tumors and provide a unique model for studies in cancer research. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  13. Spheroidal Carbonaceous Particles (SCPs) as Chronological Markers in Marine Sediments

    Science.gov (United States)

    Thornalley, D.; Rose, N.; Oppo, D.

    2016-12-01

    Spheroidal carbonaceous particles (SCPs) are a component of fly-ash, the particulate by-product of industrial high-temperature combustion of coal and fuel-oil that is released to the atmosphere with flue-gases. They are morphologically distinct and have no natural sources making them unambiguous markers of contamination from these anthropogenic sources. In naturally accumulating archives, SCPs may be used as a chronological tool as they provide a faithful record of industrial emissions and deposition. While the timing of the first presence of SCP in the 19th century, and the observed sub-surface peak are dependent on factors such as sediment accumulation rates and local industrial history, a rapid increase in SCP inputs in the mid-20thcentury appears to be a global signal corresponding to an acceleration in global electricity demand following the Second World War and the use of fuel-oil in electricity production at an industrial scale for the first time. While this approach has been widely used in lake sediments, it has not been applied to marine sediments, although there is great potential. Improved dating of 19th-20th century marine sediments has particular relevance for developing reconstructions of recent multi-decadal climate and ocean variability, and for studies that aim to place 20thcentury climate change within the context of the last millennium. Here, we present data from three sediment cores from the continental slope south of Iceland to demonstrate the temporal and spatial replicability of the SCP record in the marine environment and compare these data with cores taken from more contaminated areas off the coast of the eastern United States. The improved age model constraints provided by the analysis of SCPs has enabled a more accurate assessment of the timing of recent abrupt climate events recorded in these archives and has thus improved our understanding of likely causal climate mechanisms.

  14. A KINEMATIC STUDY OF THE ANDROMEDA DWARF SPHEROIDAL SYSTEM

    Energy Technology Data Exchange (ETDEWEB)

    Collins, Michelle L. M.; Martin, Nicolas F. [Max-Planck-Institut fuer Astronomie, Koenigstuhl 17, D-69117 Heidelberg (Germany); Chapman, Scott C.; Irwin, Michael J. [Institute of Astronomy, Madingley Rise, Cambridge CB3 0HA (United Kingdom); Rich, R. Michael [Department of Physics and Astronomy, University of California, Los Angeles, CA 90095-1547 (United States); Ibata, Rodrigo A. [Observatoire astronomique de Strasbourg, Universite de Strasbourg, CNRS, UMR 7550, 11 rue de l' Universite, F-67000 Strasbourg (France); Bate, Nicholas F.; Lewis, Geraint F. [Sydney Institute for Astronomy, School of Physics, A28, University of Sydney, NSW 2006 (Australia); Penarrubia, Jorge [Instituto de Astrofisica de Andalucia-CSIC, Glorieta de la Astronomia s/n, E-18008 Granada (Spain); Arimoto, Nobuo [Subaru Telescope, National Astronomical Observatory of Japan, 650 North A' ohoku Place, Hilo, HI 96720 (United States); Casey, Caitlin M. [Institute for Astronomy, 2680 Woodlawn Drive, Honolulu, HI 96822-1839 (United States); Ferguson, Annette M. N. [Institute for Astronomy, University of Edinburgh, Royal Observatory, Blackford Hill, Edinburgh EH9 3HJ (United Kingdom); Koch, Andreas [Zentrum fuer Astronomie der Universitaet Heidelberg, Landessternwarte, Koenigstuhl 12, D-69117 Heidelberg (Germany); McConnachie, Alan W. [NRC Herzberg Institute of Astrophysics, 5071 West Saanich Road, British Columbia, Victoria V9E 2E7 (Canada); Tanvir, Nial [Department of Physics and Astronomy, University of Leicester, University Road, Leicester LE1 7RH (United Kingdom)

    2013-05-10

    We present a homogeneous kinematic analysis of red giant branch stars within 18 of the 28 Andromeda dwarf spheroidal (dSph) galaxies, obtained using the Keck I/LRIS and Keck II/DEIMOS spectrographs. Based on their g - i colors (taken with the CFHT/MegaCam imager), physical positions on the sky, and radial velocities, we assign probabilities of dSph membership to each observed star. Using this information, the velocity dispersions, central masses, and central densities of the dark matter halos are calculated for these objects, and compared with the properties of the Milky Way dSph population. We also measure the average metallicity ([Fe/H]) from the co-added spectra of member stars for each M31 dSph and find that they are consistent with the trend of decreasing [Fe/H] with luminosity observed in the Milky Way population. We find that three of our studied M31 dSphs appear as significant outliers in terms of their central velocity dispersion, And XIX, XXI, and XXV, all of which have large half-light radii ({approx}> 700 pc) and low velocity dispersions ({sigma}{sub v} < 5 km s{sup -1}). In addition, And XXV has a mass-to-light ratio within its half-light radius of just [M/L]{sub half}=10.3{sup +7.0}{sub -6.7}, making it consistent with a simple stellar system with no appreciable dark matter component within its 1{sigma} uncertainties. We suggest that the structure of the dark matter halos of these outliers have been significantly altered by tides.

  15. A KINEMATIC STUDY OF THE ANDROMEDA DWARF SPHEROIDAL SYSTEM

    International Nuclear Information System (INIS)

    Collins, Michelle L. M.; Martin, Nicolas F.; Chapman, Scott C.; Irwin, Michael J.; Rich, R. Michael; Ibata, Rodrigo A.; Bate, Nicholas F.; Lewis, Geraint F.; Peñarrubia, Jorge; Arimoto, Nobuo; Casey, Caitlin M.; Ferguson, Annette M. N.; Koch, Andreas; McConnachie, Alan W.; Tanvir, Nial

    2013-01-01

    We present a homogeneous kinematic analysis of red giant branch stars within 18 of the 28 Andromeda dwarf spheroidal (dSph) galaxies, obtained using the Keck I/LRIS and Keck II/DEIMOS spectrographs. Based on their g – i colors (taken with the CFHT/MegaCam imager), physical positions on the sky, and radial velocities, we assign probabilities of dSph membership to each observed star. Using this information, the velocity dispersions, central masses, and central densities of the dark matter halos are calculated for these objects, and compared with the properties of the Milky Way dSph population. We also measure the average metallicity ([Fe/H]) from the co-added spectra of member stars for each M31 dSph and find that they are consistent with the trend of decreasing [Fe/H] with luminosity observed in the Milky Way population. We find that three of our studied M31 dSphs appear as significant outliers in terms of their central velocity dispersion, And XIX, XXI, and XXV, all of which have large half-light radii (∼> 700 pc) and low velocity dispersions (σ v –1 ). In addition, And XXV has a mass-to-light ratio within its half-light radius of just [M/L] half =10.3 +7.0 -6.7 , making it consistent with a simple stellar system with no appreciable dark matter component within its 1σ uncertainties. We suggest that the structure of the dark matter halos of these outliers have been significantly altered by tides.

  16. Altered interactions between unicellular and multicellular genes drive hallmarks of transformation in a diverse range of solid tumors.

    Science.gov (United States)

    Trigos, Anna S; Pearson, Richard B; Papenfuss, Anthony T; Goode, David L

    2017-06-13

    Tumors of distinct tissues of origin and genetic makeup display common hallmark cellular phenotypes, including sustained proliferation, suppression of cell death, and altered metabolism. These phenotypic commonalities have been proposed to stem from disruption of conserved regulatory mechanisms evolved during the transition to multicellularity to control fundamental cellular processes such as growth and replication. Dating the evolutionary emergence of human genes through phylostratigraphy uncovered close association between gene age and expression level in RNA sequencing data from The Cancer Genome Atlas for seven solid cancers. Genes conserved with unicellular organisms were strongly up-regulated, whereas genes of metazoan origin were primarily inactivated. These patterns were most consistent for processes known to be important in cancer, implicating both selection and active regulation during malignant transformation. The coordinated expression of strongly interacting multicellularity and unicellularity processes was lost in tumors. This separation of unicellular and multicellular functions appeared to be mediated by 12 highly connected genes, marking them as important general drivers of tumorigenesis. Our findings suggest common principles closely tied to the evolutionary history of genes underlie convergent changes at the cellular process level across a range of solid cancers. We propose altered activity of genes at the interfaces between multicellular and unicellular regions of human gene regulatory networks activate primitive transcriptional programs, driving common hallmark features of cancer. Manipulation of cross-talk between biological processes of different evolutionary origins may thus present powerful and broadly applicable treatment strategies for cancer.

  17. Magnetohydrodynamic equilibrium and stability of spheromak with spheroidal plasma-vacuum interface

    International Nuclear Information System (INIS)

    Kaneko, Shobu; Kamitani, Atsushi; Takimoto, Akio.

    1985-05-01

    The analytic solutions to the Grad-Shafranov equation are obtained for a prolate and an oblate spheroidal plasma by using Hill's vortex model. Effects of a toroidal magnetic field Bsub(phi) on the MHD equilibrium configurations are investigated by using these analytic solutions. When Bsub(phi) is larger than that of the force-free configuration, the spheroidal plasmas in a vacuum magnetic field are shown to be unable in the MHD equilibrium. The several physical quantities on the equilibrium configuration are evaluated. The spheromak plasma is proved to be unstable if dp/d psi not equal 0 and d 2 V/d psi 2 >= 0 on the magnetic axis. Here p is the pressure and V(psi) the volume surrounded by a magnetic surface of psi=const. The equilibrium configurations of the spheroidal plasmas by using Hill's vortex model are shown to satisfy the above conditions, i.e., to be unstable. (author)

  18. Magnetohydrodynamic equilibrium and stability of spheromak with spheroidal plasma-vacuum interface

    International Nuclear Information System (INIS)

    Kaneko, Shobu; Kamitani, Atsushi; Takimoto, Akio

    1985-01-01

    The analytic solutions to the Grad-Shafranov equation are obtained for a prolate and an oblate spheroidal plasma by using Hill's vortex model. Effects of a toroidal magnetic field Bsub(phi) on the MHD equilibrium configurations are investigated by using these analytic solutions. When Bsub(phi) is stronger than that of the force-free configuration, the spheroidal plasmas in a vacuum magnetic field are shown to be unable in the MHD equilibrium. The several physical quantities on the equilibrium configuration are evaluated. The spheromak plasma is proved to be unstable if dp/d psi not equal 0 and d 2 V/d psi 2 >= 0 on the magnetic axis. Here p is the pressure and V(psi) the volume surrounded by a magnetic surface of psi = const. The equilibrium configurations of the spheroidal plasmas by using Hill's vortex model are shown to satisfy the above conditions, i.e., to be unstable. (author)

  19. Scaffold-Free Coculture Spheroids of Human Colonic Adenocarcinoma Cells and Normal Colonic Fibroblasts Promote Tumorigenicity in Nude Mice

    Directory of Open Access Journals (Sweden)

    Jong-il Park

    2016-02-01

    Full Text Available The aim of this study was to form a scaffold-free coculture spheroid model of colonic adenocarcinoma cells (CACs and normal colonic fibroblasts (NCFs and to use the spheroids to investigate the role of NCFs in the tumorigenicity of CACs in nude mice. We analysed three-dimensional (3D scaffold-free coculture spheroids of CACs and NCFs. CAC Matrigel invasion assays and tumorigenicity assays in nude mice were performed to examine the effect of NCFs on CAC invasive behaviour and tumorigenicity in 3D spheroids. We investigated the expression pattern of fibroblast activation protein-α (FAP-α by immunohistochemical staining. CAC monocultures did not form densely-packed 3D spheroids, whereas cocultured CACs and NCFs formed 3D spheroids. The 3D coculture spheroids seeded on a Matrigel extracellular matrix showed higher CAC invasiveness compared to CACs alone or CACs and NCFs in suspension. 3D spheroids injected into nude mice generated more and faster-growing tumors compared to CACs alone or mixed suspensions consisting of CACs and NCFs. FAP-α was expressed in NCFs-CACs cocultures and xenograft tumors, whereas monocultures of NCFs or CACs were negative for FAP-α expression. Our findings provide evidence that the interaction between CACs and NCFs is essential for the tumorigenicity of cancer cells as well as for tumor propagation.

  20. Fully-resolved prolate spheroids in turbulent channel flows: A lattice Boltzmann study

    Science.gov (United States)

    Eshghinejadfard, Amir; Hosseini, Seyed Ali; Thévenin, Dominique

    2017-09-01

    Particles are present in many natural and industrial multiphase flows. In most practical cases, particle shape is not spherical, leading to additional difficulties for numerical studies. In this paper, DNS of turbulent channel flows with finite-size prolate spheroids is performed. The geometry includes a straight wall-bounded channel at a frictional Reynolds number of 180 seeded with particles. Three different particle shapes are considered, either spheroidal (aspect ratio λ =2 or 4) or spherical (λ =1 ). Solid-phase volume fraction has been varied between 0.75% and 1.5%. Lattice Boltzmann method (LBM) is used to model the fluid flow. The influence of the particles on the flow field is simulated by immersed boundary method (IBM). In this Eulerian-Lagrangian framework, the trajectory of each particle is computed individually. All particle-particle and particle-fluid interactions are considered (four-way coupling). Results show that, in the range of examined volume fractions, mean fluid velocity is reduced by addition of particles. However, velocity reduction by spheroids is much lower than that by spheres; 2% and 1.6%, compared to 4.6%. Maximum streamwise velocity fluctuations are reduced by addition of particle. By comparing particle and fluid velocities, it is seen that spheroids move faster than the fluid before reaching the same speed in the channel center. Spheres, on the other hand, move slower than the fluid in the buffer layer. Close to the wall, all particle types move faster than the fluid. Moreover, prolate spheroids show a preferential orientation in the streamwise direction, which is stronger close to the wall. Far from the wall, the orientation of spheroidal particles tends to isotropy.

  1. Fully-resolved prolate spheroids in turbulent channel flows: A lattice Boltzmann study

    Directory of Open Access Journals (Sweden)

    Amir Eshghinejadfard

    2017-09-01

    Full Text Available Particles are present in many natural and industrial multiphase flows. In most practical cases, particle shape is not spherical, leading to additional difficulties for numerical studies. In this paper, DNS of turbulent channel flows with finite-size prolate spheroids is performed. The geometry includes a straight wall-bounded channel at a frictional Reynolds number of 180 seeded with particles. Three different particle shapes are considered, either spheroidal (aspect ratio λ=2 or 4 or spherical (λ=1. Solid-phase volume fraction has been varied between 0.75% and 1.5%. Lattice Boltzmann method (LBM is used to model the fluid flow. The influence of the particles on the flow field is simulated by immersed boundary method (IBM. In this Eulerian-Lagrangian framework, the trajectory of each particle is computed individually. All particle-particle and particle-fluid interactions are considered (four-way coupling. Results show that, in the range of examined volume fractions, mean fluid velocity is reduced by addition of particles. However, velocity reduction by spheroids is much lower than that by spheres; 2% and 1.6%, compared to 4.6%. Maximum streamwise velocity fluctuations are reduced by addition of particle. By comparing particle and fluid velocities, it is seen that spheroids move faster than the fluid before reaching the same speed in the channel center. Spheres, on the other hand, move slower than the fluid in the buffer layer. Close to the wall, all particle types move faster than the fluid. Moreover, prolate spheroids show a preferential orientation in the streamwise direction, which is stronger close to the wall. Far from the wall, the orientation of spheroidal particles tends to isotropy.

  2. Radiation survival of cells from spheroids grown in different oxygen concentrations

    International Nuclear Information System (INIS)

    Franko, A.J.; Sutherland, R.M.

    1979-01-01

    The position of the internal, chronically hypoxic cells in spheroids was varied by alterations in the oxygen concentration in the growth medium. Such alterations were expected to cause large changes in the size of the radiobiologically hypoxic fraction. This was tested by growing and irradiating spheroids in oxygen concentrations between 5 and 20.3%, ensuring that the irradiation and growth conditions were as similar as possible. The survival curves appeared to be linear below a surviving fraction of 3 x 10 -2 , and the slopes were intermediate between the slopes of control curves for cells from spheroids irradiated in nitrogen or when fully oxygenated. Thus direct estimates of the hypoxic fractions could not be made. Two models of oxygen diffusion might explain the data. One model assumes that a large fraction of cells was fully hypoxic (radiobiologically) and that these internal, G 1 -confined, chronically hypoxic cells had a lower inherent radioresistance than the outer proliferating cells. Evidence was presented which indicated that this model was unlikely to be correct. The other model assumes that the inherent radioresistance was equal throughout the spheroid, and that the innermost cells died before the oxygen concentration was reduced sufficiently to cause full hypoxic protection. Theoretical survival curves based on this model were generated using the measured geometries ofthe spheroids and multitarget single-hit survival theory. Acceptable agreement with the postulate that the innermost cells of spheroids die at between 0.2 and 0.4% oxygen was obtained. These data may have implications regarding the relative contributions of chronic and acute hypoxia to the fraction of hypoxic cells in tumors

  3. Supermassive Black Holes and their Host Spheroids III. The Mbh-nsph Correlation

    Science.gov (United States)

    Savorgnan, Giulia A. D.

    2016-04-01

    The Sérsic {R}1/n model is the best approximation known to date for describing the light distribution of stellar spheroidal and disk components, with the Sérsic index n providing a direct measure of the central radial concentration of stars. The Sérsic index of a galaxy’s spheroidal component, nsph, has been shown to tightly correlate with the mass of the central supermassive black hole, MBH. The {M}{BH}{--}{n}{sph} correlation is also expected from other two well known scaling relations involving the spheroid luminosity, Lsph: the {L}{sph}{--}{n}{sph} and the {M}{BH}{--}{L}{sph}. Obtaining an accurate estimate of the spheroid Sérsic index requires a careful modeling of a galaxy’s light distribution and some studies have failed to recover a statistically significant {M}{BH}{--}{n}{sph} correlation. With the aim of re-investigating the {M}{BH}{--}{n}{sph} and other black hole mass scaling relations, we performed a detailed (I.e., bulge, disks, bars, spiral arms, rings, halo, nucleus, etc.) decomposition of 66 galaxies, with directly measured black hole masses, that had been imaged at 3.6 μm with Spitzer. In this paper, the third of this series, we present an analysis of the {L}{sph}{--}{n}{sph} and {M}{BH}{--}{n}{sph} diagrams. While early-type (elliptical+lenticular) and late-type (spiral) galaxies split into two separate relations in the {L}{sph}{--}{n}{sph} and {M}{BH}{--}{L}{sph} diagrams, they reunite into a single {M}{BH}\\propto {n}{sph}3.39+/- 0.15 sequence with relatively small intrinsic scatter (ɛ ≃ 0.25 {dex}). The black hole mass appears to be closely related to the spheroid central concentration of stars, which mirrors the inner gradient of the spheroid gravitational potential.

  4. Transplantation of cord blood mesenchymal stem cells as spheroids enhances vascularization.

    Science.gov (United States)

    Bhang, Suk Ho; Lee, Seahyoung; Shin, Jung-Youn; Lee, Tae-Jin; Kim, Byung-Soo

    2012-10-01

    Despite promising results from the therapeutic use of stem cells for treating ischemic diseases, the poor survival of cells transplanted into ischemic regions is one of the major problems that undermine the efficacy of stem cell therapy. Cord blood mononuclear cells (CBMNCs) are an alternative source of mesenchymal stem cells (MSCs) without disadvantages, such as the painful and invasive harvesting procedure, of MSCs derived from bone marrow or adipose tissue. In the present study, we investigated whether the angiogenic efficacy of cord blood mesenchymal stem cells (CBMSCs) can be enhanced by grafting as spheroids in a mouse hindlimb ischemia model. Human CBMSC (hCBMSC) spheroids were prepared by using the hanging-drop method. Mouse hindlimb ischemia was induced by excising the femoral artery and its branches. After surgery, the animals were divided into no-treatment, dissociated hCBMSC, and spheroid hCBMSC groups (n=8 per group) and received corresponding hCBMSC treatments. After surgery, the ischemic hindlimbs were monitored for 4 weeks, and then, the ischemic hindlimb muscles were harvested for histological analysis. Apoptotic signaling, angiogenesis-related signal pathways, and blood vessel formation were investigated in vitro and/or in vivo. The transplantation of hCBMSCs as spheroids into mouse ischemic hindlimbs significantly improved the survival of the transplanted cells by suppressing apoptotic signaling while activating antiapoptotic signaling. Furthermore, the transplantation of hCBMSCs as spheroids significantly increased the number of microvessels and smooth muscle α-actin-positive vessels in the ischemic limbs of mice, and attenuated limb loss and necrosis. Human CBMNC can be considered an alternative source of MSC, and spheroid-based hCBMSC delivery can be considered a simple and effective strategy for enhancing the therapeutic efficacy of hCBMSCs.

  5. Buildings exposed to fire

    International Nuclear Information System (INIS)

    1987-01-01

    The 24 lectures presented to the colloquium cover the following subject fields: (1) Behaviour of structural components exposed to fire; (2) Behaviour of building materials exposed to fire; (3) Thermal processes; (4) Safety related, theoretical studies. (PW) [de

  6. Theoretical investigation of resonance frequencies in long wavelength electromagnetic wave scattering process from plasma prolate and oblate spheroids placed in a dielectric layer

    Science.gov (United States)

    Ahmadizadeh, Y.; Jazi, B.; Abdoli-Arani, A.

    2014-01-01

    Response of a prolate spheroid plasma and/or an oblate spheroid plasma in presence of long wavelength electromagnetic wave has been studied. The resonance frequencies of these objects are obtained and it is found that they reduce to the resonance frequency of spherical cold plasma. Moreover, the resonant frequencies of prolate spheroid plasma and oblate spheroid plasma covered by a dielectric are investigated as well. Furthermore, their dependency on dielectric permittivity and geometry dimensions is simulated.

  7. Engineering fibrin hydrogels to promote the wound healing potential of mesenchymal stem cell spheroids.

    Science.gov (United States)

    Murphy, Kaitlin C; Whitehead, Jacklyn; Zhou, Dejie; Ho, Steve S; Leach, J Kent

    2017-12-01

    Mesenchymal stem cells (MSCs) secrete endogenous factors such as vascular endothelial growth factor (VEGF) and prostaglandin E2 (PGE 2 ) that promote angiogenesis, modulate the inflammatory microenvironment, and stimulate wound repair, and MSC spheroids secrete more trophic factors than dissociated, individual MSCs. Compared to injection of cells alone, transplantation of MSCs in a biomaterial can enhance their wound healing potential by localizing cells at the defect site and upregulating trophic factor secretion. To capitalize on the therapeutic potential of spheroids, we engineered a fibrin gel delivery vehicle to simultaneously enhance the proangiogenic and anti-inflammatory potential of entrapped human MSC spheroids. We used multifactorial statistical analysis to determine the interaction between four input variables derived from fibrin gel synthesis on four output variables (gel stiffness, gel contraction, and secretion of VEGF and PGE 2 ). Manipulation of the four input variables tuned fibrin gel biophysical properties to promote the simultaneous secretion of VEGF and PGE 2 by entrapped MSC spheroids while maintaining overall gel integrity. MSC spheroids in stiffer gels secreted the most VEGF, while PGE 2 secretion was highest in more compliant gels. Simultaneous VEGF and PGE 2 secretion was greatest using hydrogels with intermediate mechanical properties, as small increases in stiffness increased VEGF secretion while maintaining PGE 2 secretion by entrapped spheroids. The fibrin gel formulation predicted to simultaneously increase VEGF and PGE 2 secretion stimulated endothelial cell proliferation, enhanced macrophage polarization, and promoted angiogenesis when used to treat a wounded three-dimensional human skin equivalent. These data demonstrate that a statistical approach is an effective strategy to formulate fibrin gel formulations that enhance the wound healing potential of human MSCs. Mesenchymal stem cells (MSCs) are under investigation for wound

  8. Relationship of clonogenic cells and 'tumour-rescuing cells', modelled in irradiated spheroids in vitro

    International Nuclear Information System (INIS)

    Moore, J.V.; Hendry, J.H.

    1984-01-01

    Using the method of double negative logs (Gilbert, 1974), in which the probability of death (Pm) of a structure (e.g. spheroid, tumour or organism) after a given dose D, is related to the survival characteristics after irradiation of target cells (TRC) within the structure, the authors have reexamined the data of Durand (1975) for spheroids of V79-171 Chinese hamster cells grown in spinner culture, and of Pourreau-Schneider and Malaise (1981) for Na II human melanoma grown on agar. (U.K.)

  9. Composition of the spheroidal objects in KhN77TYuR-VD alloy

    International Nuclear Information System (INIS)

    Kotkis, M.A.; Nabutovskii, L.S.; Ostrov, A.E.; Zil'berman, A.G.

    1986-01-01

    The authors make an element analysis of the spheroidal objects in KhN77TYuR-VD alloy with the use of the energy dispersion microanalyzer with which the Stereoscan S-180 scanning electron microscope is equipped. Examples of the qualitative element analysis are shown. The results of the investigations show that the composition of the spheroidal inclusions includes nickel, chromium, titanium, iron, and also silicon and sulfur. The information obtained makes it possible to make an assumption on the mechanism of origin of these objects

  10. Stability of force-free spheromak plasma in spheroidal flux conserver

    International Nuclear Information System (INIS)

    Kaneko, Shobu; Tsutsui, Hiroaki

    1988-01-01

    The Woltjer-Taylor method is applied to spheromak plasmas in spheroidal flux conservers. As models of the flux conserver, both oblate and prolate spheroidal vessels with a center conductor are used. The plasma is not assumed to be nearly spherical, and the Rayleigh-Ritz method and the finite element method are used to evaluate the eigenvalues. The oblate spheromak is shown to be stable irrespective of the shape of the flux conserver. Though the prolate spheromak is unstable if there is no center conductor, it can be stable if the center conductor is installed. (author)

  11. Stability of spheroidal spheromak plasma by use of force-free approximation

    International Nuclear Information System (INIS)

    Kaneko, Shobu; Tsutsui, Hiroaki.

    1987-09-01

    The Woltjer-Taylor method is applied to spheromak plasmas in spheroidal flux conservers. As models of the flux conserver, both oblate and prolate spheroidal vessels with a center conductor are used. The plasma is not assumed to be nearly spherical, and the Rayleigh-Ritz method and the finite element method are used to evaluate the eigenvalues. The oblate spheromak is shown to be stable irrespective of the shape of the flux conserver. Though the prolate spheromak is unstable if there is no center conductor, it can be stable if the center conductor is installed. (author)

  12. A possible formation scenario for dwarf spheroidal galaxies - III. Adding star formation histories to the fiducial model

    Science.gov (United States)

    Alarcón Jara, A. G.; Fellhauer, M.; Matus Carrillo, D. R.; Assmann, P.; Urrutia Zapata, F.; Hazeldine, J.; Aravena, C. A.

    2018-02-01

    Dwarf spheroidal galaxies are regarded as the basic building blocks in the formation of larger galaxies and are the most dark matter dominated systems in the Universe, known so far. There are several models that attempt to explain their formation and evolution, but they have problems modelling the formation of isolated dwarf spheroidal galaxies. Here, we will explain a possible formation scenario in which star clusters form inside the dark matter halo of a dwarf spheroidal galaxy. These star clusters suffer from low star formation efficiency and dissolve while orbiting inside the dark matter halo. Thereby, they build the faint luminous components that we observe in dwarf spheroidal galaxies. In this paper, we study this model by adding different star formation histories to the simulations and compare the results with our previous work and observational data to show that we can explain the formation of dwarf spheroidal galaxies.

  13. Spatial distribution of elements in the spheroids by prostate tumor cells using synchrotron radiation x-ray fluorescence

    Energy Technology Data Exchange (ETDEWEB)

    Leitao, Roberta G.; Santos, Carlos Antonio N.; Junior, Antonio Palumbo; Souza, Pedro A. V. R.; Canellas, Catarine G. L.; Anjos, Marcelino J.; Nasciutti, Luiz E.; Lopes, Ricardo T. [Laboratorio de Instrumentacao Nuclear, PEN/COPPE, Universidade Federal do Rio de Janeiro, Ilha do Fundao, 21941-972, Rio de Janeiro, RJ (Brazil); Laboratorio de Biotecnologia - Bioengenharia - DIPRO, Instituto Nacional de Metrologia, Normalizacao e Qualidade Industrial, Xerem. 25250-020, Duque de Caxias, RJ (Brazil); Laboratorio de Interacoes Celulares, ICB-CCS, Universidade Federal do Rio de Janeiro, Ilha do Fundao, 21941- 590, Rio de Janeiro, RJ (Brazil); Laboratorio de Instrumentacao Nuclear, PEN/COPPE, Universidade Federal do Rio de Janeiro, Ilha do Fundao, 21941-972, Rio de Janeiro, RJ (Brazil); Laboratorio de Interacoes Celulares, ICB-CCS, Universidade Federal do Rio de Janeiro, Ilha do Fundao, 21941- 590, Rio de Janeiro, RJ (Brazil); Laboratorio de Instrumentacao Nuclear, PEN/COPPE, Universidade Federal do Rio de Janeiro, Ilha do Fundao, 21941-972, Rio de Janeiro, RJ (Brazil)

    2012-05-17

    The formation of three-dimensional cell microspheres such as spheroids has attracted attention as a useful culture technique. In this study, we investigated the trace elemental distribution (mapping) in spheroids derived from tissue prostate cancer (PCa). The measurements were performed in standard geometry of 45 deg. incidence, exciting with a white beam and using an optical capillary with 20 {mu}m diameter collimation in the XRF beam line at the Synchrotron Light National Laboratory (Campinas, Brazil). The results showed that most elements analyzed presented non-uniform distribution. P, S and Cl showed similar elemental distribution in all the samples analyzed. K, Ca, Fe, and Cu showed different elemental distribution for the spheroids analyzed. Zinc presented more intense distributions in the spheroid central region for all spheroids analyzed.

  14. Spatial distribution of elements in the spheroids by prostate tumor cells using synchrotron radiation X-ray fluorescence

    International Nuclear Information System (INIS)

    Leitao, Roberta G.; Canellas, Catarine G.L.; Anjos, Marcelino J.; Lopes, Ricardo T.; Santos, Carlos Antonio N.; Palumbo Junior, Antonio; Souza, Pedro A.V.R.; Nasciutti, Luiz E.

    2011-01-01

    The formation of three-dimensional cell microspheres such as spheroids has attracted attention as a useful culture technique. In this study, we investigated the trace elemental distribution (mapping) in spheroids derived from tissue prostate cancer (PCa). The measurements were performed in standard geometry of 45 deg incidence, exciting with a white beam and using an optical capillary with 20 μm diameter collimation in the XRF beam line at the Synchrotron Light National Laboratory (Campinas, Brazil). The results showed that most elements analyzed presented non-uniform distribution. P, S and Cl showed similar elemental distribution in all the samples analyzed. K, Ca, Fe, and Cu showed different elemental distribution for the spheroids analyzed. Zinc presented more intense distributions in the spheroid central region for all spheroids analyzed. (author)

  15. Spatial distribution of elements in the spheroids by prostate tumor cells using synchrotron radiation x-ray fluorescence

    International Nuclear Information System (INIS)

    Leitao, Roberta G.; Santos, Carlos Antonio N.; Junior, Antonio Palumbo; Souza, Pedro A. V. R.; Canellas, Catarine G. L.; Anjos, Marcelino J.; Nasciutti, Luiz E.; Lopes, Ricardo T.

    2012-01-01

    The formation of three-dimensional cell microspheres such as spheroids has attracted attention as a useful culture technique. In this study, we investigated the trace elemental distribution (mapping) in spheroids derived from tissue prostate cancer (PCa). The measurements were performed in standard geometry of 45 deg. incidence, exciting with a white beam and using an optical capillary with 20 μm diameter collimation in the XRF beam line at the Synchrotron Light National Laboratory (Campinas, Brazil). The results showed that most elements analyzed presented non-uniform distribution. P, S and Cl showed similar elemental distribution in all the samples analyzed. K, Ca, Fe, and Cu showed different elemental distribution for the spheroids analyzed. Zinc presented more intense distributions in the spheroid central region for all spheroids analyzed.

  16. The increase in radioresistance of Chinese hamster cells cultured as spheroids is correlated to changes in nuclear morphology

    International Nuclear Information System (INIS)

    Gordon, D.J.; Milner, A.E.; Beaney, R.P.; Grdina, D.J.; Vaughan, A.T.

    1990-01-01

    Chinese hamster V79 cells grown as spheroids in roller culture are more radioresistant than those grown as monolayers. The supercoiled structure of chromatin, as salt-extracted nucleoids, has been examined using flow cytometry. Irradiated viable cells from spheroid culture contain restraints to supercoil relaxation that are absent in monolayer cells. Further analysis of the chromatin organization from each growth form shows that the radioresistant spheroid cells contain a DNA-protein matrix that is more resistant to detergent-induced degradation. The increase in structural integrity may be due to the retention of a 55-60 kDa protein that is apparent in the nucleoids of spheroid, but not monolayer cells. The increase in structural integrity of the spheroid cells may explain their greater radioresistance by providing a more stable platform for high-fidelity DNA damage repair

  17. Spatial distribution of elements in the spheroids by prostate tumor cells using synchrotron radiation X-ray fluorescence

    Energy Technology Data Exchange (ETDEWEB)

    Leitao, Roberta G.; Canellas, Catarine G.L.; Anjos, Marcelino J.; Lopes, Ricardo T. [Universidade Federal do Rio de Janeiro (PEN/COPPE/UFRJ), RJ (Brazil). Coordenacao dos Programas de Pos-Graduacao de Engenharia. Programa de Energia Nuclear; Santos, Carlos Antonio N. [Instituto Nacional de Metrologia, Normalizacao e Qualidade Industrial (INMETRO), Duque de Caxias, RJ (Brazil). Lab. de Biotecnologia - Bioengenharia; Palumbo Junior, Antonio; Souza, Pedro A.V.R.; Nasciutti, Luiz E., E-mail: nasciutt@ufrj.b [Universidade Federal do Rio de Janeiro (UFRJ), RJ (Brazil). Lab. de Interacoes Celulares

    2011-07-01

    The formation of three-dimensional cell microspheres such as spheroids has attracted attention as a useful culture technique. In this study, we investigated the trace elemental distribution (mapping) in spheroids derived from tissue prostate cancer (PCa). The measurements were performed in standard geometry of 45 deg incidence, exciting with a white beam and using an optical capillary with 20 {mu}m diameter collimation in the XRF beam line at the Synchrotron Light National Laboratory (Campinas, Brazil). The results showed that most elements analyzed presented non-uniform distribution. P, S and Cl showed similar elemental distribution in all the samples analyzed. K, Ca, Fe, and Cu showed different elemental distribution for the spheroids analyzed. Zinc presented more intense distributions in the spheroid central region for all spheroids analyzed. (author)

  18. Phase field study of interfacial diffusion-driven spheroidization in a composite comprized of two mutually insoluble phases

    Energy Technology Data Exchange (ETDEWEB)

    Tian, Liang [Ames Laboratory; Russell, Alan [Ames Laboratory

    2014-03-27

    The phase field approach is a powerful computational technique to simulate morphological and microstructural evolution at the mesoscale. Spheroidization is a frequently observed morphological change of mesoscale heterogeneous structures during annealing. In this study, we used the diffuse interface phase field method to investigate the interfacial diffusion-driven spheroidization of cylindrical rod structures in a composite comprised of two mutually insoluble phases in a two-dimensional case. Perturbation of rod radius along a cylinder's axis has long been known to cause the necessary chemical potential gradient that drives spheroidization of the rod by Lord Rayleigh's instability theory. This theory indicates that a radius perturbation wavelength larger than the initial rod circumference would lead to cylindrical spheroidization. We investigated the effect of perturbation wavelength, interfacial energy, volume diffusion, phase composition, and interfacial percentage on the kinetics of spheroidization. The results match well with both the Rayleigh's instability criterion and experimental observations.

  19. Penetration and binding of monoclonal antibody in human osteosarcoma multicell spheroids. Comparison of confocal laser scanning microscopy and autoadiography

    International Nuclear Information System (INIS)

    Hjelstuen, M.H.; Rasch-Halvorsen, K.; Brekken, C.; Bruland, Oe.; Davies, C. de L.

    1996-01-01

    Penetration and binding of monoclonal antibody (MAb) in multicell osteosarcoma spheroids have been studied by autoradiography and confocal laser scanning microscopy (CLSM). Optical sectioning of the 3-dimensional spheroids was performed by CLSM. Owing to attenuation of fluorescence intensity, FITC-labelled MAb could not be detected at depths greater than 60 μm within the spheroids. The antibody uptake seen in autoradiographs and CLSM images 60 μm within the spheroids were essentially identical. MAb had reached all parts of the spheroids within 6 h. Quantitative measurements of the fluorescence intensity of FITC-labelled MAb seen in confocal images and measurements of MAb bound per cell using flow cytometry, showed that maximum uptake was reached after 6 h. The possibility to perform both quantatitive and qualitative measurements makes CLSM a promising method for studying antibody uptake in thick tissue samples. (orig.)

  20. A Duo of Potassium-Responsive Histidine Kinases Govern the Multicellular Destiny of Bacillus subtilis.

    Science.gov (United States)

    Grau, Roberto R; de Oña, Paula; Kunert, Maritta; Leñini, Cecilia; Gallegos-Monterrosa, Ramses; Mhatre, Eisha; Vileta, Darío; Donato, Verónica; Hölscher, Theresa; Boland, Wilhelm; Kuipers, Oscar P; Kovács, Ákos T

    2015-07-07

    Multicellular biofilm formation and surface motility are bacterial behaviors considered mutually exclusive. However, the basic decision to move over or stay attached to a surface is poorly understood. Here, we discover that in Bacillus subtilis, the key root biofilm-controlling transcription factor Spo0A~Pi (phosphorylated Spo0A) governs the flagellum-independent mechanism of social sliding motility. A Spo0A-deficient strain was totally unable to slide and colonize plant roots, evidencing the important role that sliding might play in natural settings. Microarray experiments plus subsequent genetic characterization showed that the machineries of sliding and biofilm formation share the same main components (i.e., surfactin, the hydrophobin BslA, exopolysaccharide, and de novo-formed fatty acids). Sliding proficiency was transduced by the Spo0A-phosphorelay histidine kinases KinB and KinC. We discovered that potassium, a previously known inhibitor of KinC-dependent biofilm formation, is the specific sliding-activating signal through a thus-far-unnoticed cytosolic domain of KinB, which resembles the selectivity filter sequence of potassium channels. The differential expression of the Spo0A~Pi reporter abrB gene and the different levels of the constitutively active form of Spo0A, Sad67, in Δspo0A cells grown in optimized media that simultaneously stimulate motile and sessile behaviors uncover the spatiotemporal response of KinB and KinC to potassium and the gradual increase in Spo0A~Pi that orchestrates the sequential activation of sliding, followed by sessile biofilm formation and finally sporulation in the same population. Overall, these results provide insights into how multicellular behaviors formerly believed to be antagonistic are coordinately activated in benefit of the bacterium and its interaction with the host. Alternation between motile and sessile behaviors is central to bacterial adaptation, survival, and colonization. However, how is the collective

  1. Modeling photopolarimetric characteristics of comet dust as a polydisperse mixture of polyshaped rough spheroids

    Science.gov (United States)

    Kolokolova, L.; Das, H.; Dubovik, O.; Lapyonok, T.

    2013-12-01

    It is widely recognized now that the main component of comet dust is aggregated particles that consist of submicron grains. It is also well known that cometary dust obey a rather wide size distribution with abundant particles whose size reaches dozens of microns. However, numerous attempts of computer simulation of light scattering by comet dust using aggregated particles have not succeeded to consider particles larger than a couple of microns due to limitations in the memory and speed of available computers. Attempts to substitute aggregates by polydisperse solid particles (spheres, spheroids, cylinders) could not consistently reproduce observed angular and spectral characteristics of comet brightness and polarization even in such a general case as polyshaped (i.e. containing particles of a variety of aspect ratios) mixture of spheroids (Kolokolova et al., In: Photopolarimetry in Remote Sensing, Kluwer Acad. Publ., 431, 2004). In this study we are checking how well cometary dust can be modeled using modeling tools for rough spheroids. With this purpose we use the software package described in Dubovik et al. (J. Geophys. Res., 111, D11208, doi:10.1029/2005JD006619d, 2006) that allows for a substantial reduction of computer time in calculating scattering properties of spheroid mixtures by means of using pre-calculated kernels - quadrature coefficients employed in the numerical integration of spheroid optical properties over size and shape. The kernels were pre-calculated for spheroids of 25 axis ratios, ranging from 0.3 to 3, and 42 size bins within the size parameter range 0.01 - 625. This software package has been recently expanded with the possibility of simulating not only smooth but also rough spheroids that is used in present study. We consider refractive indexes of the materials typical for comet dust: silicate, carbon, organics, and their mixtures. We also consider porous particles accounting on voids in the spheroids through effective medium approach. The

  2. Switching from a unicellular to multicellular organization in an Aspergillus niger hypha.

    Science.gov (United States)

    Bleichrodt, Robert-Jan; Hulsman, Marc; Wösten, Han A B; Reinders, Marcel J T

    2015-03-03

    Pores in fungal septa enable cytoplasmic streaming between hyphae and their compartments. Consequently, the mycelium can be considered unicellular. However, we show here that Woronin bodies close ~50% of the three most apical septa of growing hyphae of Aspergillus niger. The incidence of closure of the 9th and 10th septa was even ≥94%. Intercompartmental streaming of photoactivatable green fluorescent protein (PA-GFP) was not observed when the septa were closed, but open septa acted as a barrier, reducing the mobility rate of PA-GFP ~500 times. This mobility rate decreased with increasing septal age and under stress conditions, likely reflecting a regulatory mechanism affecting septal pore diameter. Modeling revealed that such regulation offers effective control of compound concentration between compartments. Modeling also showed that the incidence of septal closure in A. niger had an even stronger impact on cytoplasmic continuity. Cytoplasm of hyphal compartments was shown not to be in physical contact when separated by more than 4 septa. Together, data show that apical compartments of growing hyphae behave unicellularly, while older compartments have a multicellular organization. The hyphae of higher fungi are compartmentalized by porous septa that enable cytosolic streaming. Therefore, it is believed that the mycelium shares cytoplasm. However, it is shown here that the septa of Aspergillus niger are always closed in the oldest part of the hyphae, and therefore, these compartments are physically isolated from each other. In contrast, only part of the septa is closed in the youngest part of the hyphae. Still, compartments in this hyphal part are physically isolated when separated by more than 4 septa. Even open septa act as a barrier for cytoplasmic mixing. The mobility rate through such septa reduces with increasing septal age and under stress conditions. Modeling shows that the septal pore width is set such that its regulation offers maximal control of

  3. A molecular timescale of eukaryote evolution and the rise of complex multicellular life

    Science.gov (United States)

    Hedges, S. Blair; Blair, Jaime E.; Venturi, Maria L.; Shoe, Jason L.

    2004-01-01

    BACKGROUND: The pattern and timing of the rise in complex multicellular life during Earth's history has not been established. Great disparity persists between the pattern suggested by the fossil record and that estimated by molecular clocks, especially for plants, animals, fungi, and the deepest branches of the eukaryote tree. Here, we used all available protein sequence data and molecular clock methods to place constraints on the increase in complexity through time. RESULTS: Our phylogenetic analyses revealed that (i) animals are more closely related to fungi than to plants, (ii) red algae are closer to plants than to animals or fungi, (iii) choanoflagellates are closer to animals than to fungi or plants, (iv) diplomonads, euglenozoans, and alveolates each are basal to plants+animals+fungi, and (v) diplomonads are basal to other eukaryotes (including alveolates and euglenozoans). Divergence times were estimated from global and local clock methods using 20-188 proteins per node, with data treated separately (multigene) and concatenated (supergene). Different time estimation methods yielded similar results (within 5%): vertebrate-arthropod (964 million years ago, Ma), Cnidaria-Bilateria (1,298 Ma), Porifera-Eumetozoa (1,351 Ma), Pyrenomycetes-Plectomycetes (551 Ma), Candida-Saccharomyces (723 Ma), Hemiascomycetes-filamentous Ascomycota (982 Ma), Basidiomycota-Ascomycota (968 Ma), Mucorales-Basidiomycota (947 Ma), Fungi-Animalia (1,513 Ma), mosses-vascular plants (707 Ma), Chlorophyta-Tracheophyta (968 Ma), Rhodophyta-Chlorophyta+Embryophyta (1,428 Ma), Plantae-Animalia (1,609 Ma), Alveolata-plants+animals+fungi (1,973 Ma), Euglenozoa-plants+animals+fungi (1,961 Ma), and Giardia-plants+animals+fungi (2,309 Ma). By extrapolation, mitochondria arose approximately 2300-1800 Ma and plastids arose 1600-1500 Ma. Estimates of the maximum number of cell types of common ancestors, combined with divergence times, showed an increase from two cell types at 2500 Ma to

  4. A molecular timescale of eukaryote evolution and the rise of complex multicellular life

    Directory of Open Access Journals (Sweden)

    Venturi Maria L

    2004-01-01

    Full Text Available Abstract Background The pattern and timing of the rise in complex multicellular life during Earth's history has not been established. Great disparity persists between the pattern suggested by the fossil record and that estimated by molecular clocks, especially for plants, animals, fungi, and the deepest branches of the eukaryote tree. Here, we used all available protein sequence data and molecular clock methods to place constraints on the increase in complexity through time. Results Our phylogenetic analyses revealed that (i animals are more closely related to fungi than to plants, (ii red algae are closer to plants than to animals or fungi, (iii choanoflagellates are closer to animals than to fungi or plants, (iv diplomonads, euglenozoans, and alveolates each are basal to plants+animals+fungi, and (v diplomonads are basal to other eukaryotes (including alveolates and euglenozoans. Divergence times were estimated from global and local clock methods using 20–188 proteins per node, with data treated separately (multigene and concatenated (supergene. Different time estimation methods yielded similar results (within 5%: vertebrate-arthropod (964 million years ago, Ma, Cnidaria-Bilateria (1,298 Ma, Porifera-Eumetozoa (1,351 Ma, Pyrenomycetes-Plectomycetes (551 Ma, Candida-Saccharomyces (723 Ma, Hemiascomycetes-filamentous Ascomycota (982 Ma, Basidiomycota-Ascomycota (968 Ma, Mucorales-Basidiomycota (947 Ma, Fungi-Animalia (1,513 Ma, mosses-vascular plants (707 Ma, Chlorophyta-Tracheophyta (968 Ma, Rhodophyta-Chlorophyta+Embryophyta (1,428 Ma, Plantae-Animalia (1,609 Ma, Alveolata-plants+animals+fungi (1,973 Ma, Euglenozoa-plants+animals+fungi (1,961 Ma, and Giardia-plants+animals+fungi (2,309 Ma. By extrapolation, mitochondria arose approximately 2300-1800 Ma and plastids arose 1600-1500 Ma. Estimates of the maximum number of cell types of common ancestors, combined with divergence times, showed an increase from two cell types at 2500 Ma to ~10

  5. Flames High Resolution Spectroscopy of RGB Stars in the Carina Dwarf Spheroidal Galaxy

    NARCIS (Netherlands)

    Lemasle, B.; Hill, V.; Tolstoy, E.; Venn, K.; Koleva, M; Prugniel, P; Vauglin,

    Carina is a small and faint classical dwarf spheroidal galaxy in the halo of the Milky Way with a highly episodic star formation history (e.g., Hurley-Keller et al. 1998). Using VLT/FLAMES in high resolution mode, we significantly increase the sample of stars with abundance determinations in Carina,

  6. VLT/FLAMES spectroscopy of red giant branch stars in the Fornax dwarf spheroidal galaxy

    NARCIS (Netherlands)

    Lemasle, B.; de Boer, T.J.L.; Hill, V.; Tolstoy, E.; Irwin, M.J.; Jablonka, P.; Venn, K.; Battaglia, G.; Starkenburg, E.; Shetrone, M.; Letarte, B.; François, P.; Helmi, A.; Primas, F.; Kaufer, A.; Szeifert, T.

    2014-01-01

    Context. Fornax is one of the most massive dwarf spheroidal galaxies in the Local Group. The Fornax field star population is dominated by intermediate age stars but star formation was going on over almost its entire history. It has been proposed that Fornax experienced a minor merger event. Aims.

  7. Chemical composition of extremely metal-poor stars in the Sextans dwarf spheroidal galaxy

    NARCIS (Netherlands)

    Aoki, W.; Arimoto, N.; Sadakane, K.; Tolstoy, E.; Battaglia, G.; Jablonka, P.; Shetrone, M.; Letarte, B.; Irwin, M.; Hill, V.; Francois, P.; Venn, K.; Primas, F.; Helmi, A.; Kaufer, A.; Tafelmeyer, M.; Szeifert, T.; Babusiaux, C.

    Context. Individual stars in dwarf spheroidal galaxies around the Milky Way Galaxy have been studied both photometrically and spectroscopically. Extremely metal-poor stars among them are very valuable because they should record the early enrichment in the Local Group. However, our understanding of

  8. Internal kinematics and dynamical models of dwarf spheroidal galaxies around the Milky Way

    NARCIS (Netherlands)

    Battaglia, Giuseppina; Helmi, Amina; Breddels, Maarten

    We review our current understanding of the internal dynamical properties of the dwarf spheroidal galaxies surrounding the Milky Way. These are the most dark matter dominated galaxies, and as such may be considered ideal laboratories to test the current concordance cosmological model, and in

  9. VLT/UVES abundances in four nearby dwarf spheroidal galaxies. II. Implications for understanding galaxy evolution

    NARCIS (Netherlands)

    Tolstoy, E; Venn, KA; Shetrone, M; Primas, F; Hill, [No Value; Kaufer, A; Szeifert, T

    We have used the Ultraviolet Visual-Echelle Spectrograph (UVES) on Kueyen (UT2) of the Very Large Telescope to take spectra of 15 individual red giant stars in the centers of four nearby dwarf spheroidal galaxies (dSph's) : Sculptor, Fornax, Carina, and Leo I. We measure the abundance variations of

  10. A new study of stellar substructures in the Fornax dwarf spheroidal galaxy

    NARCIS (Netherlands)

    de Boer, T. J. L.; Tolstoy, E.; Saha, A.; Olszewski, E. W.

    Using deep V, B - V wide-field photometry, we have conducted a new study of stellar over-densities in the Fornax dwarf spheroidal galaxy by determining detailed star formation histories from colour-magnitude diagram analysis. We have concentrated on the relatively young stellar component ( We have

  11. Cardiac spheroids as promising in vitro models to study the human heart microenvironment

    DEFF Research Database (Denmark)

    Polonchuk, Liudmila; Chabria, Mamta; Badi, Laura

    2017-01-01

    Three-dimensional in vitro cell systems are a promising alternative to animals to study cardiac biology and disease. We have generated three-dimensional in vitro models of the human heart ("cardiac spheroids", CSs) by co-culturing human primary or iPSC-derived cardiomyocytes, endothelial cells an...

  12. Three-dimensional spheroid culture promotes odonto/osteoblastic differentiation of dental pulp cells.

    Science.gov (United States)

    Yamamoto, Mioko; Kawashima, Nobuyuki; Takashino, Nami; Koizumi, Yu; Takimoto, Koyo; Suzuki, Noriyuki; Saito, Masahiro; Suda, Hideaki

    2014-03-01

    Three-dimensional (3D) spheroid culture is a method for creating 3D aggregations of cells and their extracellular matrix without a scaffold mimicking the actual tissues. The aim of this study was to evaluate the effects of 3D spheroid culture on the phenotype of immortalized mouse dental papilla cells (MDPs) that have the ability to differentiate into odontoblasts. We cultured MDPs for 1, 3, 7, and 14 days in 96-well low-attachment culture plates for 3D spheroid culture or flat-bottomed plates for two-dimensional (2D) monolayer culture. Cell proliferation and apoptosis were detected by immunohistochemical staining of Ki67 and cleaved caspase-3, respectively. Hypoxia was measured by the hypoxia probe LOX-1. Odonto/osteoblastic differentiation marker gene expression was evaluated by quantitative PCR. We also determined mineralized nodule formation, alkaline phosphatase (ALP) activity, and dentine matrix protein-1 (DMP1) expression. Vinculin and integrin signalling-related proteins were detected immunohistochemically. Odonto/osteoblastic marker gene expression and mineralized nodule formation were significantly up-regulated in 3D spheroid-cultured MDPs compared with those in 2D monolayer-cultured MDPs (podonto/osteoblastic differentiation of MDPs, which may be mediated by integrin signalling. Copyright © 2013 Elsevier Ltd. All rights reserved.

  13. Can physical stellar collisions explain the blue stragglers in the dwarf spheroidal galaxies?

    International Nuclear Information System (INIS)

    Leonard, P.J.T.

    1993-01-01

    The hypothesis that the blue stragglers in the dwarf spheroidal galaxie have a collisional origin is considered. If all of the dark matter in these galaxies is in the form of low-mass stars and the binary frequency is ≅ 50%, then it is quite possible that ≅ 10% to 20% of their blue stragglers have been produced by physical stellar collisions

  14. Three-dimensional in vitro cancer spheroid models for Photodynamic Therapy: Strengths and Opportunities

    Science.gov (United States)

    Evans, Conor

    2015-03-01

    Three dimensional, in vitro spheroid cultures offer considerable utility for the development and testing of anticancer photodynamic therapy regimens. More complex than monolayer cultures, three-dimensional spheroid systems replicate many of the important cell-cell and cell-matrix interactions that modulate treatment response in vivo. Simple enough to be grown by the thousands and small enough to be optically interrogated, spheroid cultures lend themselves to high-content and high-throughput imaging approaches. These advantages have enabled studies investigating photosensitizer uptake, spatiotemporal patterns of therapeutic response, alterations in oxygen diffusion and consumption during therapy, and the exploration of mechanisms that underlie therapeutic synergy. The use of quantitative imaging methods, in particular, has accelerated the pace of three-dimensional in vitro photodynamic therapy studies, enabling the rapid compilation of multiple treatment response parameters in a single experiment. Improvements in model cultures, the creation of new molecular probes of cell state and function, and innovations in imaging toolkits will be important for the advancement of spheroid culture systems for future photodynamic therapy studies.

  15. Stem Cell Spheroids and Ex Vivo Niche Modeling: Rationalization and Scaling-Up.

    Science.gov (United States)

    Chimenti, Isotta; Massai, Diana; Morbiducci, Umberto; Beltrami, Antonio Paolo; Pesce, Maurizio; Messina, Elisa

    2017-04-01

    Improved protocols/devices for in vitro culture of 3D cell spheroids may provide essential cues for proper growth and differentiation of stem/progenitor cells (S/PCs) in their niche, allowing preservation of specific features, such as multi-lineage potential and paracrine activity. Several platforms have been employed to replicate these conditions and to generate S/PC spheroids for therapeutic applications. However, they incompletely reproduce the niche environment, with partial loss of its highly regulated network, with additional hurdles in the field of cardiac biology, due to debated resident S/PCs therapeutic potential and clinical translation. In this contribution, the essential niche conditions (metabolic, geometric, mechanical) that allow S/PCs maintenance/commitment will be discussed. In particular, we will focus on both existing bioreactor-based platforms for the culture of S/PC as spheroids, and on possible criteria for the scaling-up of niche-like spheroids, which could be envisaged as promising tools for personalized cardiac regenerative medicine, as well as for high-throughput drug screening.

  16. Electromagnetic waves in irregular multilayered spheroidal structures of finite conductivity: full wave solutions

    International Nuclear Information System (INIS)

    Bahar, E.

    1976-01-01

    The propagation of electromagnetic waves excited by electric dipoles oriented along the axis of multilayered spheroidal structures of finite conductivity is investigated. The electromagnetic parameters and the thickness of the layers of the structure are assumed to be functions of the latitude. In the analysis, electric and magnetic field transforms that constitute a discrete and a continuous spectrum of spherical waves are used to provide a suitable basis for the expansion of the electromagnetic fields at any point in the irregular spheroidal structure. For spheroidal structures with good conducting cores, the terms in the solutions associated with the continuous part of the wave spectrum vanish. In general, however, when the skin depth for the core is large compared to its dimensions or when the sources are located in the core of the structure and propagation in the core is of special interest, the contribution from the continuous part of the wave spectrum cannot be neglected. At each interface between the layers of the irregular spheroidal structure, exact boundary conditions are imposed. Since the terms of the field expansions in the irregular structure do not individually satisfy the boundary conditions, Maxwell's equations are reduced to sets of coupled ordinary first-order differential equations for the wave amplitudes. The solutions are shown to satisfy the reciprocity relationships in electromagnetic theory. The analysis may be applied to problems of radio wave propagation in a nonuniform model of the earth-ionosphere waveguide, particularly when focusing effects at the antipodes are important

  17. Ground eigenvalue and eigenfunction of a spin-weighted spheroidal wave equation in low frequencies

    Institute of Scientific and Technical Information of China (English)

    Tang Wen-Lin; Tian Gui-Hua

    2011-01-01

    Spin-weighted spheroidal wave functions play an important role in the study of the linear stability of rotating Kerr black holes and are studied by the perturbation method in supersymmetric quantum mechanics. Their analytic ground eigenvalues and eigenfunctions are obtained by means of a series in low frequency. The ground eigenvalue and eigenfunction for small complex frequencies are numerically determined.

  18. Detailed Studies of the Sculptor Dwarf Spheroidal Galaxy in the Milky Way halo

    NARCIS (Netherlands)

    Tolstoy, Eline

    In and around the Milky Way halo there are a number of low mass low luminosity dwarf galaxies. Several of these systems have been studied in great detail. I describe recent photometric and spectroscopic studies of the Sculptor dwarf spheroidal galaxy made as part of the DART survey of nearby dwarf

  19. Effect of Cu Salt Molarity on the Nanostructure of CuO Prolate Spheroid

    Science.gov (United States)

    Sabeeh, Sabah H.; Hussein, Hashim Abed; Judran, Hadia Kadhim

    Copper sulfate pentahydrate was used as a source of Cu ion with five different molarities (0.02, 0.05, 0.1, 0.15, 2 and 0.25M). XRD, FE-SEM and TEM techniques all showed that CuO samples have polycrystalline monoclinic structure. CuO prolate spheroid is assembled from nanoparticles as building units. It was demonstrated that the purity, morphology, size range of prolate spheroid and density of nano building units are significantly influenced by Cu precursor’s molarity. The pure phase of CuO prolate spheroid was produced via molarity of 0.2M with crystallite size of 15.1565nm while the particle size of building units ranges from 16nm to 21nm. The stability of CuO nanosuspension or nanofluid was evaluated by zeta potential analysis. The obtained properties of specific structure with large surface area of CuO prolate spheroid make it a promising candidate for wide range of potential applications as in nanofluids for cooling purposes.

  20. Dynamics of Small Inertia-Free Spheroidal Particles in a Turbulent Channel Flow

    Science.gov (United States)

    Challabotla, Niranjan Reddy; Zhao, Lihao; Andersson, Helge I.; Department of Energy; Process Engineering Team

    2015-11-01

    The study of small non-spherical particles suspended in turbulent fluid flows is of interest in view of the potential applications in industry and the environment. In the present work, we investigated the dynamics of inertia-free spheroidal particles suspended in fully-developed turbulent channel flow at Re τ = 180 by using the direct numerical simulations (DNS) for the Eulerian fluid phase coupled with the Lagrangian point-particle tracking. We considered inertia-free spheroidal particles with a wide range of aspect ratios from 0.01 to 50, i.e. from flat disks to long rods. Although the spheroids passively translate along with the fluid, the particle orientation and rotation strongly depend on the particle shape. The flattest disks were preferentially aligned with their symmetry axis normal to the wall, whereas the longest rods aligned parallel to the wall. Strong mean rotational spin was observed for spherical particles and this has been damped with increasing asphericity both for rod-like and disk-like spheroids. The anisotropic mean and fluctuating fluid vorticity resulted in particle spin anisotropies which exhibited a complex dependence on the particle asphericty. The Research Council of Norway, Notur and COST Action FP1005 are gratefully acknowledged.

  1. Influence of boron on ferrite formation in copper-added spheroidal graphite cast iron

    Directory of Open Access Journals (Sweden)

    Ying Zou

    2014-07-01

    Full Text Available This paper reviews the original work of the authors published recently, describing the influence of B on the matrix of the Cuadded spheroidal graphite cast iron. The effect of Cu has been corrected as a ferrite formation promoter in the matrix of the grey cast iron by the usage of high-purity material. Also, this paper focuses on the ferrite formation and the observation of the Cu distribution in the B-added and B-free Cu-containing spheroidal graphite cast iron. The Cu film on the spheroidal graphite can be successfully observed in the B-free sample using a special etching method. However, in the B-added sample, no Cu film could be found, while the secondary graphite was formed on the surface of the spheroidal graphite. The interaction between B and Cu is stressed as a peculiar phenomenon by the employment of a contrast experiment of B and Mn. The heat treatment could make Cu precipitate more significantly in the eutectic cells and in the matrix in the form of large Cu particles because of the limited solubility of Cu.

  2. Porous Nb-Ti based alloy produced from plasma spheroidized powder

    Science.gov (United States)

    Li, Qijun; Zhang, Lin; Wei, Dongbin; Ren, Shubin; Qu, Xuanhui

    Spherical Nb-Ti based alloy powder was prepared by the combination of plasma spheroidization and mechanical alloying. Phase constituents, microstructure and surface state of the powder, and pore characteristics of the resulting porous alloy were investigated. The results show that the undissolved W and V in the mechanically alloyed powder is fully alloyed after spheroidization, and single β phase is achieved. Particle size of the spheroidized powder is in the range of 20-110 μm. With the decrease of particle size, a transformation from typical dendrite solidification structure to fine cell microstructure occurs. The surface of the spheroidized powder is coated by a layer of oxides consisting mainly of TiO2 and Nb2O5. Probabilities of sinter-neck formation and particle coalescence increases with increasing sintering temperature. Porous skeleton with relatively homogeneous pore distribution and open pore channel is formed after vacuum sintering at 1700 °C, and the porosity is 32%. The sintering kinetic analysis indicates that grain boundary diffusion is the primary mass transport mechanism during sintering process.

  3. Numerical Simulation of Spheroidization Process of TiAl Alloy Powders in Radio Frequency Plasma

    Directory of Open Access Journals (Sweden)

    ZHU Langping

    2017-06-01

    Full Text Available A numerical simulation method was used to study the radio frequency plasma spheroidization process of TiAl alloy powder. The effects of velocity field and temperature field on the motion trajectory and mass change of TiAl alloy powder with different particle size were analyzed.The results show that the temperature of powder particles increases rapidly under high temperature plasma, surface evaporation cause the reduction of particle size, and particles with small size tend to evaporate quickly. The motion trajectory of particles with different sizes in the lower end of the cooling tube is different obviously, small particles tend to enter the air outlet,while the larger particles are easy to fall down to the bottom of the cooling tube to be collected. Increasing air flow rate can improve the velocity of air flow in the spheroidizing system, causing larger particles to be taken away by the air, resulting in yield reduction. The simulation results of TiAl alloy powder spheroidization are close to the experimental results refer to parameters such as powder size distribution, average particle size and powder yield, and the model is in good accordance with the actual process of the spheroidization.

  4. THE STELLAR SPHEROID, THE DISK, AND THE DYNAMICS OF THE COSMIC WEB

    International Nuclear Information System (INIS)

    Domínguez-Tenreiro, R.; Obreja, A.; Brook, C. B.; Martínez-Serrano, F. J.; Serna, A.; Stinson, G.

    2015-01-01

    Models of the advanced stages of gravitational instability predict that baryons that form the stellar populations of current galaxies at z = 0 displayed a web-like structure at high z, as part of the cosmic web (CW). We explore details of these predictions using cosmological hydrodynamical simulations. When the stellar populations of the spheroid and disk components of simulated late-type galaxies are traced back separately to high zs we found CW-like structures where spheroid progenitors are more evolved than disk progenitors. The distinction between the corresponding stellar populations, as driven by their specific angular momentum content j, can be explained in terms of the CW evolution, extended to two processes occurring at lower z. First, the spheroid progenitors strongly lose j at collapse, which contrasts with the insignificant j loss of the disk progenitors. The second is related to the lack of alignment, at assembly, between the spheroid-to-be material and the already settled proto-disk, in contrast to the alignment of disk-to-be material, in some cases resulting from circumgalactic, disk-induced gravitational torques. The different final outcomes of these low-z processes have their origins in the different initial conditions driven by the CW dynamics

  5. Photometric analyses of abundances in dwarf spheroidal galaxies and globular clusters

    International Nuclear Information System (INIS)

    Light, R.M.

    1988-01-01

    This study investigated the abundance characteristics of several dwarf spheroidal galaxies. The chemical properties of stars in these galaxies are tracers of the origin and evolution of their stellar populations, and thus can provide constraints on the theories of their formation. To derive this abundance information, photometric observations of stars in a sample of globular clusters, covering a large range in metallicity, were analyzed. Parameters describing the position of the red giant branch were found to correlate very well with cluster metallicity over a large range in abundance. These measurements, made in the Thuan-Gunn photometry system, provide ranking schemes which are, with accurate photometry, more sensitive to changes in metallicity than similar broadband BV parameters. The relations were used to derive an improved estimate of the metallicity of cluster NGC 5053. These metallicity relations were used to analyze the Thuan-Gunn system photometry produced for the Sculptor, Ursa Minor, and Carina galaxies. The excellent agreement between their metallicities and those from other previous studies indicates that globular cluster red giant branch parameters are very useful in ranking dwarf spheroidal populations by metallicity. Together with other galaxian data, strong correlations can be seen between the mean metallicities and dispersions in metallicity and the luminosities of the dwarf spheroidal galaxies. These trends also seem to apply to members of the dwarf elliptical class of galaxies. The ramifications that these correlations and the existence of a metallicity gradient in Sculptor have on the formation of the dwarf spheroidals are discussed

  6. Impact of a small ellipticity on the sustainability condition of developed turbulence in a precessing spheroid

    Science.gov (United States)

    Horimoto, Yasufumi; Simonet-Davin, Gabriel; Katayama, Atsushi; Goto, Susumu

    2018-04-01

    We experimentally investigate the flow transition to developed turbulence in a precessing spheroid with a small ellipticity. Fully developed turbulence appears through a subcritical transition when we fix the Reynolds number (the spin rate) and gradually increase the Poincaré number (the precession rate). In the transitional range of the Poincaré number, two qualitatively different turbulent states (i.e., fully developed turbulence and quiescent turbulence with a spin-driven global circulation) are stable and they are connected by a hysteresis loop. This discontinuous transition is in contrast to the continuous transition in a precessing sphere, for which neither bistable turbulent states nor hysteresis loops are observed. The small ellipticity of the container makes the global circulation of the confined fluid more stable, and it requires much stronger precession of the spheroid, than a sphere, for fully developed turbulence to be sustained. Nevertheless, once fully developed turbulence is sustained, its flow structures are almost identical in the spheroid and sphere. The argument [Lorenzani and Tilgner, J. Fluid Mech. 492, 363 (2003), 10.1017/S002211200300572X; Noir et al., Geophys. J. Int. 154, 407 (2003), 10.1046/j.1365-246X.2003.01934.x] on the basis of the analytical solution [Busse, J. Fluid Mech. 33, 739 (1968), 10.1017/S0022112068001655] of the steady global circulation in a weak precession range well describes the onset of the fully developed turbulence in the spheroid.

  7. Asymmetric segregation of damaged cellular components in spatially structured multicellular organisms.

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    Charlotte Strandkvist

    Full Text Available The asymmetric distribution of damaged cellular components has been observed in species ranging from fission yeast to humans. To study the potential advantages of damage segregation, we have developed a mathematical model describing ageing mammalian tissue, that is, a multicellular system of somatic cells that do not rejuvenate at cell division. To illustrate the applicability of the model, we specifically consider damage incurred by mutations to mitochondrial DNA, which are thought to be implicated in the mammalian ageing process. We show analytically that the asymmetric distribution of damaged cellular components reduces the overall damage level and increases the longevity of the cell population. Motivated by the experimental reports of damage segregation in human embryonic stem cells, dividing symmetrically with respect to cell-fate, we extend the model to consider spatially structured systems of cells. Imposing spatial structure reduces, but does not eliminate, the advantage of asymmetric division over symmetric division. The results suggest that damage partitioning could be a common strategy for reducing the accumulation of damage in a wider range of cell types than previously thought.

  8. The number of cell types, information content, and the evolution of complex multicellularity

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    Karl J. Niklas

    2014-12-01

    Full Text Available The number of different cell types (NCT characterizing an organism is often used to quantify organismic complexity. This method results in the tautology that more complex organisms have a larger number of different kinds of cells, and that organisms with more different kinds of cells are more complex. This circular reasoning can be avoided (and simultaneously tested when NCT is plotted against different measures of organismic information content (e.g., genome or proteome size. This approach is illustrated by plotting the NCT of representative diatoms, green and brown algae, land plants, invertebrates, and vertebrates against data for genome size (number of base-pairs, proteome size (number of amino acids, and proteome functional versatility (number of intrinsically disordered protein domains or residues. Statistical analyses of these data indicate that increases in NCT fail to keep pace with increases in genome size, but exceed a one-to-one scaling relationship with increasing proteome size and with increasing numbers of intrinsically disordered protein residues. We interpret these trends to indicate that comparatively small increases in proteome (and not genome size are associated with disproportionate increases in NCT, and that proteins with intrinsically disordered domains enhance cell type diversity and thus contribute to the evolution of complex multicellularity.

  9. Cellular and functional specificity among ferritin-like proteins in the multicellular cyanobacterium Nostoc punctiforme.

    Science.gov (United States)

    Ekman, Martin; Sandh, Gustaf; Nenninger, Anja; Oliveira, Paulo; Stensjö, Karin

    2014-03-01

    Ferritin-like proteins constitute a remarkably heterogeneous protein family, including ferritins, bacterioferritins and Dps proteins. The genome of the filamentous heterocyst-forming cyanobacterium Nostoc punctiforme encodes five ferritin-like proteins. In the present paper, we report a multidimensional characterization of these proteins. Our phylogenetic and bioinformatics analyses suggest both structural and physiological differences among the ferritin-like proteins. The expression of these five genes responded differently to hydrogen peroxide treatment, with a significantly higher rise in transcript level for Npun_F3730 as compared with the other four genes. A specific role for Npun_F3730 in the cells tolerance against hydrogen peroxide was also supported by the inactivation of Npun_F3730, Npun_R5701 and Npun_R6212; among these, only the ΔNpun_F3730 strain showed an increased sensitivity to hydrogen peroxide compared with wild type. Analysis of promoter-GFP reporter fusions of the ferritin-like genes indicated that Npun_F3730 and Npun_R5701 were expressed in all cell types of a diazotrophic culture, while Npun_F6212 was expressed specifically in heterocysts. Our study provides the first comprehensive analysis combining functional differentiation and cellular specificity within this important group of proteins in a multicellular cyanobacterium. © 2013 John Wiley & Sons Ltd and Society for Applied Microbiology.

  10. Theoretical analysis of the spatio-temporal structure of bone multicellular units

    International Nuclear Information System (INIS)

    Buenzli, P R; Pivonka, P; Gardiner, B S; Smith, D W; Dunstan, C R; Mundy, G R

    2010-01-01

    Bone multicellular units (BMUs) maintain the viability of the skeletal tissue by coordinating locally the sequence of bone resorption and bone formation performed by cells of the osteoclastic and osteoblastic lineage. Understanding the emergence and the net bone balance of such structured microsystems out of the complex network of biochemical interactions between bone cells is fundamental for many bone-related diseases and the evaluation of fracture risk. Based on current experimental knowledge, we propose a spatio-temporal continuum model describing the interactions of osteoblastic and osteoclastic cells. We show that this model admits travelling-wave-like solutions with well-confined cell profiles upon specifying external conditions mimicking the environment encountered in cortical bone remodelling. The shapes of the various cell concentration profiles within this travelling structure are intrinsically linked to the parameters of the model such as differentiation, proliferation, and apoptosis rates of bone cells. The internal structure of BMUs is reproduced, allowing for experimental calibration. The spatial distribution of the key regulatory factors can also be exhibited, which in diseased states could give hints as to the biochemical agent most accountable for the disorder.

  11. Agent-based model of angiogenesis simulates capillary sprout initiation in multicellular networks.

    Science.gov (United States)

    Walpole, J; Chappell, J C; Cluceru, J G; Mac Gabhann, F; Bautch, V L; Peirce, S M

    2015-09-01

    Many biological processes are controlled by both deterministic and stochastic influences. However, efforts to model these systems often rely on either purely stochastic or purely rule-based methods. To better understand the balance between stochasticity and determinism in biological processes a computational approach that incorporates both influences may afford additional insight into underlying biological mechanisms that give rise to emergent system properties. We apply a combined approach to the simulation and study of angiogenesis, the growth of new blood vessels from existing networks. This complex multicellular process begins with selection of an initiating endothelial cell, or tip cell, which sprouts from the parent vessels in response to stimulation by exogenous cues. We have constructed an agent-based model of sprouting angiogenesis to evaluate endothelial cell sprout initiation frequency and location, and we have experimentally validated it using high-resolution time-lapse confocal microscopy. ABM simulations were then compared to a Monte Carlo model, revealing that purely stochastic simulations could not generate sprout locations as accurately as the rule-informed agent-based model. These findings support the use of rule-based approaches for modeling the complex mechanisms underlying sprouting angiogenesis over purely stochastic methods.

  12. Study of the tensile properties of individual multicellular fibres generated by Bacillus subtilis

    Science.gov (United States)

    Ye, Xuan; Zhao, Liang; Liang, Jiecun; Li, Xide; Chen, Guo-Qiang

    2017-04-01

    Multicellular fibres formed by Bacillus subtilis (B. subtilis) are attracting interest because of their potential application as degradable biomaterials. However, mechanical properties of individual fibres remain unknown because of their small dimensions. Herein, a new approach is developed to investigate the tensile properties of individual fibres with an average diameter of 0.7 μm and a length range of 25.7-254.3 μm. Variations in the tensile strengths of fibres are found to be the result of variable interactions among pairs of microbial cells known as septa. Using Weibull weakest-link model to study this mechanical variability, we predict the length effect of the sample. Moreover, the mechanical properties of fibres are found to depend highly on relative humidity (RH), with a brittle-ductile transition occurring around RH = 45%. The elastic modulus is 5.8 GPa in the brittle state, while decreases to 62.2 MPa in the ductile state. The properties of fibres are investigated by using a spring model (RH  45%) for the time-dependent response. Loading-unloading experiments and numerical calculations demonstrate that necking instability comes from structural changes (septa) and viscoelasticity dominates the deformation of fibres at high RH.

  13. Recombination every day: abundant recombination in a virus during a single multi-cellular host infection.

    Directory of Open Access Journals (Sweden)

    Remy Froissart

    2005-03-01

    Full Text Available Viral recombination can dramatically impact evolution and epidemiology. In viruses, the recombination rate depends on the frequency of genetic exchange between different viral genomes within an infected host cell and on the frequency at which such co-infections occur. While the recombination rate has been recently evaluated in experimentally co-infected cell cultures for several viruses, direct quantification at the most biologically significant level, that of a host infection, is still lacking. This study fills this gap using the cauliflower mosaic virus as a model. We distributed four neutral markers along the viral genome, and co-inoculated host plants with marker-containing and wild-type viruses. The frequency of recombinant genomes was evaluated 21 d post-inoculation. On average, over 50% of viral genomes recovered after a single host infection were recombinants, clearly indicating that recombination is very frequent in this virus. Estimates of the recombination rate show that all regions of the genome are equally affected by this process. Assuming that ten viral replication cycles occurred during our experiment-based on data on the timing of coat protein detection-the per base and replication cycle recombination rate was on the order of 2 x 10(-5 to 4 x 10(-5. This first determination of a virus recombination rate during a single multi-cellular host infection indicates that recombination is very frequent in the everyday life of this virus.

  14. Tissues development in stems of Aristolochia clematitis L. in the point of view of multicellular complexes formation

    Directory of Open Access Journals (Sweden)

    Zofia Puławska

    2014-01-01

    Full Text Available After cytokinesis the cells do not separate but remain within the wall of the mother cell. After a series of divisions a multicellular complex arises. In the stems of Aristolochia clematitis procambium is closer related to protoxylem than to protophloem, and metaphloem is closer related to metaxylem than to protophloem. Since protophloem has a closer common origin with fibre primordia than with the remaining tissues, it cannot be decided unequivocally what is the origin of the fibres or when procambium differentiates. The common origin of the primary vascular tissues is visible in the pattern of the multicellular complexes, whereas the common origin of the secondary vascular tissue developing in the underground several-year-old parts of the stem can be traced in the arrangement of the single radial tiers. Some characteristics of symplastic growth are discussed.

  15. Design of Online Spheroidization Process for 1.0C-1.5Cr Bearing Steel and Microstructure Analysis

    Science.gov (United States)

    Li, Zhen-Xing; Li, Chang-Sheng; Ren, Jin-Yi; Li, Bin-Zhou; Suh, Dong-Woo

    2018-02-01

    Using thermo-mechanical control process, the online spheroidization annealing process of 1.0C-1.5Cr bearing steel was designed. Apart from intercritical online spheroidization (IS), a novel subcritical online spheroidization (SS) process was proposed, which is characterized by water-cooling to around 773 K (500 °C) after the final rolling pass, and then directly reheating to 973 K (700 °C) for isothermal holding. Compared with the results from the traditional offline spheroidization (TS) process, the size of spheroidized carbides is similar in both the TS and IS processes, whereas it is much smaller in the SS process. After spheroidization annealing, microstructure evolution during austenitization and quenching treatment was examined. It is shown that the refining of spheroidized carbides accelerates the dissolution of carbides during the austenitizing process, and decreases the size of undissolved carbides. In addition, the SS process can obtain finer prior austenite grain after quenching, which contributes to the enhancement of final hardness.

  16. Micro-environmental mechanical stress controls tumor spheroid size and morphology by suppressing proliferation and inducing apoptosis in cancer cells.

    Directory of Open Access Journals (Sweden)

    Gang Cheng

    Full Text Available Compressive mechanical stress produced during growth in a confining matrix limits the size of tumor spheroids, but little is known about the dynamics of stress accumulation, how the stress affects cancer cell phenotype, or the molecular pathways involved.We co-embedded single cancer cells with fluorescent micro-beads in agarose gels and, using confocal microscopy, recorded the 3D distribution of micro-beads surrounding growing spheroids. The change in micro-bead density was then converted to strain in the gel, from which we estimated the spatial distribution of compressive stress around the spheroids. We found a strong correlation between the peri-spheroid solid stress distribution and spheroid shape, a result of the suppression of cell proliferation and induction of apoptotic cell death in regions of high mechanical stress. By compressing spheroids consisting of cancer cells overexpressing anti-apoptotic genes, we demonstrate that mechanical stress-induced apoptosis occurs via the mitochondrial pathway.Our results provide detailed, quantitative insight into the role of micro-environmental mechanical stress in tumor spheroid growth dynamics, and suggest how tumors grow in confined locations where the level of solid stress becomes high. An important implication is that apoptosis via the mitochondrial pathway, induced by compressive stress, may be involved in tumor dormancy, in which tumor growth is held in check by a balance of apoptosis and proliferation.

  17. Principles of the Kenzan Method for Robotic Cell Spheroid-Based Three-Dimensional Bioprinting.

    Science.gov (United States)

    Moldovan, Nicanor I; Hibino, Narutoshi; Nakayama, Koichi

    2017-06-01

    Bioprinting is a technology with the prospect to change the way many diseases are treated, by replacing the damaged tissues with live de novo created biosimilar constructs. However, after more than a decade of incubation and many proofs of concept, the field is still in its infancy. The current stagnation is the consequence of its early success: the first bioprinters, and most of those that followed, were modified versions of the three-dimensional printers used in additive manufacturing, redesigned for layer-by-layer dispersion of biomaterials. In all variants (inkjet, microextrusion, or laser assisted), this approach is material ("scaffold") dependent and energy intensive, making it hardly compatible with some of the intended biological applications. Instead, the future of bioprinting may benefit from the use of gentler scaffold-free bioassembling methods. A substantial body of evidence has accumulated, indicating this is possible by use of preformed cell spheroids, which have been assembled in cartilage, bone, and cardiac muscle-like constructs. However, a commercial instrument capable to directly and precisely "print" spheroids has not been available until the invention of the microneedles-based ("Kenzan") spheroid assembling and the launching in Japan of a bioprinter based on this method. This robotic platform laces spheroids into predesigned contiguous structures with micron-level precision, using stainless steel microneedles ("kenzans") as temporary support. These constructs are further cultivated until the spheroids fuse into cellular aggregates and synthesize their own extracellular matrix, thus attaining the needed structural organization and robustness. This novel technology opens wide opportunities for bioengineering of tissues and organs.

  18. Multi-cellular natural killer (NK) cell clusters enhance NK cell activation through localizing IL-2 within the cluster

    Science.gov (United States)

    Kim, Miju; Kim, Tae-Jin; Kim, Hye Mi; Doh, Junsang; Lee, Kyung-Mi

    2017-01-01

    Multi-cellular cluster formation of natural killer (NK) cells occurs during in vivo priming and potentiates their activation to IL-2. However, the precise mechanism underlying this synergy within NK cell clusters remains unclear. We employed lymphocyte-laden microwell technologies to modulate contact-mediated multi-cellular interactions among activating NK cells and to quantitatively assess the molecular events occurring in multi-cellular clusters of NK cells. NK cells in social microwells, which allow cell-to-cell contact, exhibited significantly higher levels of IL-2 receptor (IL-2R) signaling compared with those in lonesome microwells, which prevent intercellular contact. Further, CD25, an IL-2R α chain, and lytic granules of NK cells in social microwells were polarized toward MTOC. Live cell imaging of lytic granules revealed their dynamic and prolonged polarization toward neighboring NK cells without degranulation. These results suggest that IL-2 bound on CD25 of one NK cells triggered IL-2 signaling of neighboring NK cells. These results were further corroborated by findings that CD25-KO NK cells exhibited lower proliferation than WT NK cells, and when mixed with WT NK cells, underwent significantly higher level of proliferation. These data highlights the existence of IL-2 trans-presentation between NK cells in the local microenvironment where the availability of IL-2 is limited.

  19. Adult Lung Spheroid Cells Contain Progenitor Cells and Mediate Regeneration in Rodents With Bleomycin-Induced Pulmonary Fibrosis.

    Science.gov (United States)

    Henry, Eric; Cores, Jhon; Hensley, M Taylor; Anthony, Shirena; Vandergriff, Adam; de Andrade, James B M; Allen, Tyler; Caranasos, Thomas G; Lobo, Leonard J; Cheng, Ke

    2015-11-01

    Lung diseases are devastating conditions and ranked as one of the top five causes of mortality worldwide according to the World Health Organization. Stem cell therapy is a promising strategy for lung regeneration. Previous animal and clinical studies have focused on the use of mesenchymal stem cells (from other parts of the body) for lung regenerative therapies. We report a rapid and robust method to generate therapeutic resident lung progenitors from adult lung tissues. Outgrowth cells from healthy lung tissue explants are self-aggregated into three-dimensional lung spheroids in a suspension culture. Without antigenic sorting, the lung spheroids recapitulate the stem cell niche and contain a natural mixture of lung stem cells and supporting cells. In vitro, lung spheroid cells can be expanded to a large quantity and can form alveoli-like structures and acquire mature lung epithelial phenotypes. In severe combined immunodeficiency mice with bleomycin-induced pulmonary fibrosis, intravenous injection of human lung spheroid cells inhibited apoptosis, fibrosis, and infiltration but promoted angiogenesis. In a syngeneic rat model of pulmonary fibrosis, lung spheroid cells outperformed adipose-derived mesenchymal stem cells in reducing fibrotic thickening and infiltration. Previously, lung spheroid cells (the spheroid model) had only been used to study lung cancer cells. Our data suggest that lung spheroids and lung spheroid cells from healthy lung tissues are excellent sources of regenerative lung cells for therapeutic lung regeneration. The results from the present study will lead to future human clinical trials using lung stem cell therapies to treat various incurable lung diseases, including pulmonary fibrosis. The data presented here also provide fundamental knowledge regarding how injected stem cells mediate lung repair in pulmonary fibrosis. ©AlphaMed Press.

  20. Endothelial network formed with human dermal microvascular endothelial cells in autologous multicellular skin substitutes.

    Science.gov (United States)

    Ponec, Maria; El Ghalbzouri, Abdoelwaheb; Dijkman, Remco; Kempenaar, Johanna; van der Pluijm, Gabri; Koolwijk, Pieter

    2004-01-01

    A human skin equivalent from a single skin biopsy harboring keratinocytes and melanocytes in the epidermal compartment, and fibroblasts and microvascular dermal endothelial cells in the dermal compartment was developed. The results of the study revealed that the nature of the extracellular matrix of the dermal compartments plays an important role in establishment of endothelial network in vitro. With rat-tail type I collagen matrices only lateral but not vertical expansion of endothelial networks was observed. In contrast, the presence of extracellular matrix of entirely human origin facilitated proper spatial organization of the endothelial network. Namely, when human dermal fibroblasts and microvascular endothelial cells were seeded on the bottom of an inert filter and subsequently epidermal cells were seeded on top of it, fibroblasts produced extracellular matrix throughout which numerous branched tubes were spreading three-dimensionally. Fibroblasts also facilitated the formation of basement membrane at the epidermal/matrix interface. Under all culture conditions, fully differentiated epidermis was formed with numerous melanocytes present in the basal epidermal cell layer. The results of the competitive RT-PCR revealed that both keratinocytes and fibroblasts expressed VEGF-A, -B, -C, aFGF and bFGF mRNA, whereas fibroblasts also expressed VEGF-D mRNA. At protein level, keratinocytes produced 10 times higher amounts of VEGF-A than fibroblasts did. The generation of multicellular skin equivalent from a single human skin biopsy will stimulate further developments for its application in the treatment of full-thickness skin defects. The potential development of biodegradable, biocompatible material suitable for these purposes is a great challenge for future research.

  1. An integrated in vitro imaging platform for characterizing filarial parasite behavior within a multicellular microenvironment.

    Directory of Open Access Journals (Sweden)

    Timothy Kassis

    2014-11-01

    Full Text Available Lymphatic Filariasis, a Neglected Tropical Disease, is caused by thread-like parasitic worms, including B. malayi, which migrate to the human lymphatic system following transmission. The parasites reside in collecting lymphatic vessels and lymph nodes for years, often resulting in lymphedema, elephantiasis or hydrocele. The mechanisms driving worm migration and retention within the lymphatics are currently unknown. We have developed an integrated in vitro imaging platform capable of quantifying B. malayi migration and behavior in a multicellular microenvironment relevant to the initial site of worm injection by incorporating the worm in a Polydimethylsiloxane (PDMS microchannel in the presence of human dermal lymphatic endothelial cells (LECs and human dermal fibroblasts (HDFs. The platform utilizes a motorized controllable microscope with CO2 and temperature regulation to allow for worm tracking experiments with high resolution over large length and time scales. Using post-acquisition algorithms, we quantified four parameters: 1 speed, 2 thrashing intensity, 3 percentage of time spent in a given cell region and 4 persistence ratio. We demonstrated the utility of our system by quantifying these parameters for L3 B. malayi in the presence of LECs and HDFs. Speed and thrashing increased in the presence of both cell types and were altered within minutes upon exposure to the anthelmintic drug, tetramisole. The worms displayed no targeted migration towards either cell type for the time course of this study (3 hours. When cells were not present in the chamber, worm thrashing correlated directly with worm speed. However, this correlation was lost in the presence of cells. The described platform provides the ability to further study B. malayi migration and behavior.

  2. Generation model of positional values as cell operation during the development of multicellular organisms.

    Science.gov (United States)

    Ogawa, Ken-ichiro; Miyake, Yoshihiro

    2011-03-01

    Many conventional models have used the positional information hypothesis to explain each elementary process of morphogenesis during the development of multicellular organisms. Their models assume that the steady concentration patterns of morphogens formed in an extracellular environment have an important property of positional information, so-called "robustness". However, recent experiments reported that a steady morphogen pattern, the concentration gradient of the Bicoid protein, during early Drosophila embryonic development is not robust for embryo-to-embryo variability. These reports encourage a reconsideration of a long-standing problem in systematic cell differentiation: what is the entity of positional information for cells? And, what is the origin of the robust boundary of gene expression? To address these problems at a cellular level, in this article we pay attention to the re-generative phenomena that show another important property of positional information, "size invariance". In view of regenerative phenomena, we propose a new mathematical model to describe the generation mechanism of a spatial pattern of positional values. In this model, the positional values are defined as the values into which differentiable cells transform a spatial pattern providing positional information. The model is mathematically described as an associative algebra composed of various terms, each of which is the multiplication of some fundamental operators under the assumption that the operators are derived from the remarkable properties of cell differentiation on an amputation surface in regenerative phenomena. We apply this model to the concentration pattern of the Bicoid protein during the anterior-posterior axis formation in Drosophila, and consider the conditions needed to establish the robust boundary of the expression of the hunchback gene. Copyright © 2010 Elsevier Ireland Ltd. All rights reserved.

  3. Integrative multicellular biological modeling: a case study of 3D epidermal development using GPU algorithms

    Directory of Open Access Journals (Sweden)

    Christley Scott

    2010-08-01

    Full Text Available Abstract Background Simulation of sophisticated biological models requires considerable computational power. These models typically integrate together numerous biological phenomena such as spatially-explicit heterogeneous cells, cell-cell interactions, cell-environment interactions and intracellular gene networks. The recent advent of programming for graphical processing units (GPU opens up the possibility of developing more integrative, detailed and predictive biological models while at the same time decreasing the computational cost to simulate those models. Results We construct a 3D model of epidermal development and provide a set of GPU algorithms that executes significantly faster than sequential central processing unit (CPU code. We provide a parallel implementation of the subcellular element method for individual cells residing in a lattice-free spatial environment. Each cell in our epidermal model includes an internal gene network, which integrates cellular interaction of Notch signaling together with environmental interaction of basement membrane adhesion, to specify cellular state and behaviors such as growth and division. We take a pedagogical approach to describing how modeling methods are efficiently implemented on the GPU including memory layout of data structures and functional decomposition. We discuss various programmatic issues and provide a set of design guidelines for GPU programming that are instructive to avoid common pitfalls as well as to extract performance from the GPU architecture. Conclusions We demonstrate that GPU algorithms represent a significant technological advance for the simulation of complex biological models. We further demonstrate with our epidermal model that the integration of multiple complex modeling methods for heterogeneous multicellular biological processes is both feasible and computationally tractable using this new technology. We hope that the provided algorithms and source code will be a

  4. Anaerobic Respiration Using a Complete Oxidative TCA Cycle Drives Multicellular Swarming in Proteus mirabilis

    Science.gov (United States)

    Alteri, Christopher J.; Himpsl, Stephanie D.; Engstrom, Michael D.; Mobley, Harry L. T.

    2012-01-01

    ABSTRACT Proteus mirabilis rapidly migrates across surfaces using a periodic developmental process of differentiation alternating between short swimmer cells and elongated hyperflagellated swarmer cells. To undergo this vigorous flagellum-mediated motility, bacteria must generate a substantial proton gradient across their cytoplasmic membranes by using available energy pathways. We sought to identify the link between energy pathways and swarming differentiation by examining the behavior of defined central metabolism mutants. Mutations in the tricarboxylic acid (TCA) cycle (fumC and sdhB mutants) caused altered patterns of swarming periodicity, suggesting an aerobic pathway. Surprisingly, the wild-type strain swarmed on agar containing sodium azide, which poisons aerobic respiration; the fumC TCA cycle mutant, however, was unable to swarm on azide. To identify other contributing energy pathways, we screened transposon mutants for loss of swarming on sodium azide and found insertions in the following genes that involved fumarate metabolism or respiration: hybB, encoding hydrogenase; fumC, encoding fumarase; argH, encoding argininosuccinate lyase (generates fumarate); and a quinone hydroxylase gene. These findings validated the screen and suggested involvement of anaerobic electron transport chain components. Abnormal swarming periodicity of fumC and sdhB mutants was associated with the excretion of reduced acidic fermentation end products. Bacteria lacking SdhB were rescued to wild-type pH and periodicity by providing fumarate, independent of carbon source but dependent on oxygen, while fumC mutants were rescued by glycerol, independent of fumarate only under anaerobic conditions. These findings link multicellular swarming patterns with fumarate metabolism and membrane electron transport using a previously unappreciated configuration of both aerobic and anaerobic respiratory chain components. PMID:23111869

  5. Study of the effect of vitamins C and E on the radiation response of multicell spheroids treated with Adriamycin

    International Nuclear Information System (INIS)

    Sridhar, R.; Stroude, E.; Inch, W.R.

    1979-01-01

    Treatment with Adriamycin (0.6 μg/ml for 60 min) was not cytotoxic to multicell spheroids. At this concentration, the drug was not a sensitizer of hypoxic cells in V79 multicell spheroids, which were irradiated at 37 0 C in medium equilibrated with a mixture of 5% O 2 :3% CO 2 :92% N 2 . The addition of vitamins C and E did not increase the radiation sensitivity of Adriamycin-treated spheroids. In some experiments, catalase was included in the growth medium to overcome the toxic effect of hydrogen peroxide, which is known to be formed in solutions containing vitamin C and also in Adriamycin solutions treated with vitamin C or microsomal preparations. As a result of these experiments, it was found that catalase increased the radiation killing in multicell spheroids

  6. Solving ground eigenvalue and eigenfunction of spheroidal wave equation at low frequency by supersymmetric quantum mechanics method

    Institute of Scientific and Technical Information of China (English)

    Tang Wen-Lin; Tian Gui-Hua

    2011-01-01

    The spheroidal wave functions are found to have extensive applications in many branches of physics and mathematics. We use the perturbation method in supersymmetric quantum mechanics to obtain the analytic ground eigenvalue and the ground eigenfunction of the angular spheroidal wave equation at low frequency in a series form. Using this approach, the numerical determinations of the ground eigenvalue and the ground eigenfunction for small complex frequencies are also obtained.

  7. Expansion of a zero-order Bessel beam in spheroidal coordinates by generalized Lorenz–Mie theory

    International Nuclear Information System (INIS)

    Han, L.; Han, Y.P.; Cui, Z.W.; Wang, J.J.

    2014-01-01

    An analytic solution to the scattering of the zero-order Bessel beam by a spheroidal particle is constructed on the basis of the generalized Lorenz–Mie theory (GLMT). The spheroidal beam shape coefficients (BSCs) of the zero-order Bessel beam are directly expressed in spheroidal coordinates and computed conveniently using an intrinsic method. Utilizing the tangential continuity of the electromagnetic fields, the expression coefficients of scattered and internal fields are determined. Numerical results concerning scattered field in the far zone are displayed for various parameters of the incident electromagnetic beam and of the scatter. These results are expected to provide useful insights into the scattering of a Bessel beam by spheroidal particles and particle manipulation applications using Bessel beams. - Highlights: • The scattering of zero-order Bessel beam by dielectric spheroid is investigated. • The analytic solution is constructed within the framework of the GLMT. • The spheroidal beam shape coefficients of the zero-order Bessel beam are computed by use an intrinsic method. • Numerical results concerning scattered field are displayed for various parameters

  8. Investigating the spectral characteristics of backscattering from heterogeneous spheroidal nuclei using broadband finite-difference time-domain simulations

    Science.gov (United States)

    Chao, Guo-Shan; Sung, Kung-Bin

    2010-02-01

    Backscattered light spectra have been used to extract size distribution of cell nuclei in epithelial tissues for noninvasive detection of precancerous lesions. In existing experimental studies, size estimation is achieved by assuming nuclei as homogeneous spheres or spheroids and fitting the measured data with models based on Mie theory. However, the validity of simplifying nuclei as homogeneous spheres has not been thoroughly examined. In this study, we investigate the spectral characteristics of backscattering from models of spheroidal nuclei under plane wave illumination using three-dimensional finite-difference time-domain (FDTD) simulation. A modulated Gaussian pulse is used to obtain wavelength dependent scattering intensity with a single FDTD run. The simulated model of nuclei consists of a nucleolus and randomly distributed chromatin condensation in homogeneous cytoplasm and nucleoplasm. The results show that backscattering spectra from spheroidal nuclei have similar oscillating patterns to those from homogeneous spheres with the diameter equal to the projective length of the spheroidal nucleus along the propagation direction. The strength of backscattering is enhanced in heterogeneous spheroids as compared to homogeneous spheroids. The degree of which backscattering spectra of heterogeneous nuclei deviate from Mie theory is highly dependent on the distribution of chromatin/nucleolus but not sensitive to nucleolar size, refractive index fluctuation or chromatin density.

  9. Imaging- and Flow Cytometry-based Analysis of Cell Position and the Cell Cycle in 3D Melanoma Spheroids

    Science.gov (United States)

    Beaumont, Kimberley A.; Anfosso, Andrea; Ahmed, Farzana

    2015-01-01

    Three-dimensional (3D) tumor spheroids are utilized in cancer research as a more accurate model of the in vivo tumor microenvironment, compared to traditional two-dimensional (2D) cell culture. The spheroid model is able to mimic the effects of cell-cell interaction, hypoxia and nutrient deprivation, and drug penetration. One characteristic of this model is the development of a necrotic core, surrounded by a ring of G1 arrested cells, with proliferating cells on the outer layers of the spheroid. Of interest in the cancer field is how different regions of the spheroid respond to drug therapies as well as genetic or environmental manipulation. We describe here the use of the fluorescence ubiquitination cell cycle indicator (FUCCI) system along with cytometry and image analysis using commercial software to characterize the cell cycle status of cells with respect to their position inside melanoma spheroids. These methods may be used to track changes in cell cycle status, gene/protein expression or cell viability in different sub-regions of tumor spheroids over time and under different conditions. PMID:26779761

  10. Cell-free DNA in a three-dimensional spheroid cell culture model

    DEFF Research Database (Denmark)

    Aucamp, Janine; Calitz, Carlemi; Bronkhorst, Abel J.

    2017-01-01

    Background Investigating the biological functions of cell-free DNA (cfDNA) is limited by the interference of vast numbers of putative sources and causes of DNA release into circulation. Utilization of three-dimensional (3D) spheroid cell cultures, models with characteristics closer to the in vivo...... cultures can serve as effective, simplified in vivo-simulating “closed-circuit” models since putative sources of cfDNA are limited to only the targeted cells. In addition, cfDNA can also serve as an alternative or auxiliary marker for tracking spheroid growth, development and culture stability. Biological...... significance 3D cell cultures can be used to translate “closed-circuit” in vitro model research into data that is relevant for in vivo studies and clinical applications. In turn, the utilization of cfDNA during 3D culture research can optimize sample collection without affecting the stability of the growth...

  11. On the role of the transformation eigenstrain in the growth or shrinkage of spheroidal isotropic precipitations

    International Nuclear Information System (INIS)

    Fischer, F.D.; Boehm, H.J.

    2005-01-01

    The jumps of the strain and stress tensors on the surface of elastic homogeneous or inhomogeneous ellipsoidal inclusions embedded in an elastic matrix are obtained from results reported in the literature. They are used to derive closed-form expressions for the thermodynamic force in such matrix-inclusion systems that are subjected to a generally defined homogeneous transformation eigenstrain. A detailed study is presented for an isotropic spheroidal inclusion in an isotropic matrix in which the most important parameters are the inclusion's aspect ratio α and an eigenstrain triaxiality parameter d-bar. The fluctuations of the thermodynamic force are investigated for a set of specific transformation eigenstrain tensors and are presented for inclusion shapes ranging from disk-like to fiber-like spheroids

  12. Gravitational collision efficiency of nonspherical aerosols II: motion of an oblate spheroid in a viscous fluid

    Energy Technology Data Exchange (ETDEWEB)

    Tuttle, R.F.; Loyalka, S.K.

    1985-06-01

    The collisional dynamics of nonspherical aerosols is modeled by the introduction of a shape factor, US . Mechanistic calculation of US requires knowledge of the flow fields around the aerosols. Since actual aerosols can be complicated in shape and since the computation of flow fields can be quite difficult, insights into the nature of US are gained by using the superposition technique and studying aerosols that have tractable flow fields. The motion of an oblate spheroid in a viscous fluid is considered. The Navier-Stokes equations and associated boundary conditions are represented in oblate spheroidal coordinates. A combination of finite differences and spline-interpolation techniques is used to transform these equations to a form suitable for numerical computations. Converged results for the flow fields are obtained for a 0 to 5 range of Reynolds numbers. In the limit of zero Reynolds number, the results are found to be in agreement with the analytical solutions of Oberbeck.

  13. Generalized prolate spheroidal wave functions for optical finite fractional Fourier and linear canonical transforms.

    Science.gov (United States)

    Pei, Soo-Chang; Ding, Jian-Jiun

    2005-03-01

    Prolate spheroidal wave functions (PSWFs) are known to be useful for analyzing the properties of the finite-extension Fourier transform (fi-FT). We extend the theory of PSWFs for the finite-extension fractional Fourier transform, the finite-extension linear canonical transform, and the finite-extension offset linear canonical transform. These finite transforms are more flexible than the fi-FT and can model much more generalized optical systems. We also illustrate how to use the generalized prolate spheroidal functions we derive to analyze the energy-preservation ratio, the self-imaging phenomenon, and the resonance phenomenon of the finite-sized one-stage or multiple-stage optical systems.

  14. Angular velocity of a spheroid log rolling in a simple shear at small Reynolds number

    Science.gov (United States)

    Meibohm, Jan; Candelier, Fabien; Rosen, Tomas; Einarsson, Jonas; Lundell, Fredrik; Mehlig, Bernhard

    2016-11-01

    We analyse the angular velocity of a small neutrally buoyant spheroid log rolling in a simple shear. When the effect of fluid inertia is negligible the angular velocity ω -> equals half the fluid vorticity. We compute by singular perturbation theory how weak fluid inertia reduces the angular velocity in an unbounded shear, and how this reduction depends upon the shape of the spheroid (on its aspect ratio). In addition we determine the angular velocity by direct numerical simulations. The results are in excellent agreement with the theory at small but not too small values of the shear Reynolds number, for all aspect ratios considered. For the special case of a sphere we find ω / s = - 1 / 2 + 0 . 0540Re 3 / 2 where s is the shear rate and Re is the shear Reynolds number. This result differs from that derived by Lin et al. who obtained a numerical coefficient roughly three times larger.

  15. Suspected Perinatal Depression Revealed to be Hereditary Diffuse Leukoencephalopathy with Spheroids.

    Science.gov (United States)

    Blume, Josefine; Weissert, Robert

    2017-01-01

    Early motor symptoms of neurodegenerative diseases often appear in combination with psychiatric symptoms, such as depression or personality changes, and are in danger of being misdiagnosed as psychogenic in young patients. We present the case of a 32-year-old woman who presented with rapid-onset depression, followed by a hypokinetic movement disorder and cognitive decline during pregnancy. Genetic testing revealed a mutation in the colony-stimulating factor 1 receptor gene, which led to the diagnosis of hereditary diffuse leukoencephalopathy with spheroids. Hereditary diffuse leukoencephalopathy with spheroids (HDLS) is probably an under-recognized disease. HDLS should be considered in patients with rapidly progressing parkinsonian symptoms and dementia accompanied by white matter lesions.

  16. Suspected Perinatal Depression Revealed to be Hereditary Diffuse Leukoencephalopathy with Spheroids

    Directory of Open Access Journals (Sweden)

    Josefine Blume

    2017-01-01

    Full Text Available Early motor symptoms of neurodegenerative diseases often appear in combination with psychiatric symptoms, such as depression or personality changes, and are in danger of being misdiagnosed as psychogenic in young patients. We present the case of a 32-year-old woman who presented with rapid-onset depression, followed by a hypokinetic movement disorder and cognitive decline during pregnancy. Genetic testing revealed a mutation in the colony-stimulating factor 1 receptor gene, which led to the diagnosis of hereditary diffuse leukoencephalopathy with spheroids. Hereditary diffuse leukoencephalopathy with spheroids (HDLS is probably an under-recognized disease. HDLS should be considered in patients with rapidly progressing parkinsonian symptoms and dementia accompanied by white matter lesions.

  17. Generation of uniform magnetic field using a spheroidal helical coil structure

    International Nuclear Information System (INIS)

    Öztürk, Yavuz; Aktaş, Bekir

    2016-01-01

    Uniformity of magnetic fields are of great importance especially in magnetic resonance studies, namely in magnetic resonance spectroscopy applications (NMR, FMR, ESR, EPR etc.) and magnetic resonance imaging applications (MRI, FMRI). Field uniformity is also required in some other applications such as eddy current probes, magnetometers, magnetic traps, particle counters etc. Here we proposed a coil winding regime, which follows the surface of a spheroid (an ellipsoid of rotation); in light of previous theoretical studies suggesting perfect uniformity for a constant ampere per turn in the axial direction thereof. We demonstrated our theoretical results from finite element calculations suggesting 0.15% of field uniformity for the proposed structure, which we called a Spheroidal Helical Coil. (paper)

  18. Dielectric tensor elements for the description of waves in rotating inhomogeneous magnetized plasma spheroids

    Science.gov (United States)

    Abdoli-Arani, A.; Ramezani-Arani, R.

    2012-11-01

    The dielectric permittivity tensor elements of a rotating cold collisionless plasma spheroid in an external magnetic field with toroidal and axial components are obtained. The effects of inhomogeneity in the densities of charged particles and the initial toroidal velocity on the dielectric permittivity tensor and field equations are investigated. The field components in terms of their toroidal components are calculated and it is shown that the toroidal components of the electric and magnetic fields are coupled by two differential equations. The influence of thermal and collisional effects on the dielectric tensor and field equations in the rotating plasma spheroid are also investigated. In the limiting spherical case, the dielectric tensor of a stationary magnetized collisionless cold plasma sphere is presented.

  19. Simulation study of spheroidal dust gains charging: Applicable to dust grain alignment

    International Nuclear Information System (INIS)

    Zahed, H.; Sobhanian, S.; Mahmoodi, J.; Khorram, S.

    2006-01-01

    The charging process of nonspherical dust grains in an unmagnetized plasma as well as in the presence of a magnetic field is studied. It is shown that unlike the spherical dust grain, due to nonhomogeneity of charge distribution on the spheroidal dust surface, the resultant electric forces on electrons and ions are different. This process produces some surface charge density gradient on the nonspherical grain surface. Effects of a magnetic field and other plasma parameters on the properties of the dust particulate are studied. It has been shown that the alignment direction could be changed or even reversed with the magnetic field and plasma parameters. Finally, the charge distribution on the spheroidal grain surface is studied for different ambient parameters including plasma temperature, neutral collision frequency, and the magnitude of the magnetic field

  20. Confinement of nonneutral spheroidal plasmas in multi-ring electrode traps

    International Nuclear Information System (INIS)

    Mohri, Akihiro; Yuyama, Tetsumori; Michishita, Toshinori; Higaki, Hiroyuki; Tanaka, Hitoshi; Yamazawa, Yohei; Aoyagi, Masayuki

    1998-01-01

    A nonneutral spheroidal plasma can be settled in a rigid rotor equilibrium inside a closed conducting cell independently of induced image charges on the cell wall if the electrostatic potential distribution on the wall surface is set equal to the sum of the external hyperbolic potential (r 2 -2z 2 ) and the self-potential produced by the plasma. A confinement system equipped with a train of properly biased ring electrodes can approximately generate any axisymmetric potential, including the above field. Experiments on confinement of electron spheroids in such a system showed that the confinement time became the longest when the condition to diminish the image charge effects was satisfied. The observed frequency of the centre-of-mass harmonic oscillation of the plasma in this configuration was in good agreement with the estimated one. (author)

  1. Anodic Dissolution of Spheroidal Graphite Cast Iron with Different Pearlite Areas in Sulfuric Acid Solutions

    Directory of Open Access Journals (Sweden)

    Yoshikazu Miyata

    2013-01-01

    Full Text Available The rate equation of anodic dissolution reaction of spheroidal graphite cast iron in sulfuric acid solutions at 298 K has been studied. The cast irons have different areas of pearlite. The anodic Tafel slope of 0.043 V decade−1 and the reaction order with respect to the hydroxyl ion activity of 1 are obtained by the linear potential sweep technique. The anodic current density does not depend on the area of pearlite. There is no difference in the anodic dissolution reaction mechanisms between pure iron and spheroidal graphite cast iron. The anodic current density of the cast iron is higher than that of the pure iron.

  2. Measurement uncertainty evaluation of cellular spheroids surface tension in compressing tests using Young-Laplace equation

    Science.gov (United States)

    Beatrici, Anderson; Santos Baptista, Leandra; Mauro Granjeiro, José

    2018-03-01

    Regenerative Medicine comprises the Biotechnology, Tissue Engineering and Biometrology for stem cell therapy. Starting from stem cells extracted from the patient, autologous implant, these cells are cultured and differentiated into other tissues, for example, articular cartilage. These cells are reorganized into microspheres (cell spheroids). Such tissue units are recombined into functional tissues constructs that can be implanted in the injured region for regeneration. It is necessary the biomechanical characterization of these constructed to determine if their properties are similar to native tissue. In this study was carried out the modeling of the calculation of uncertainty of the surface tension of cellular spheroids with the use of the Young-Laplace equation. We obtained relative uncertainties about 10%.

  3. Curcumin-loaded chitosan-cholesterol micelles: evaluation in monolayers and 3D cancer spheroid model.

    Science.gov (United States)

    Muddineti, Omkara Swami; Kumari, Preeti; Ray, Eupa; Ghosh, Balaram; Biswas, Swati

    2017-06-02

    To improve the bioavailability and anticancer potential of curcumin by using a cholesterol-conjugated chitosan micelle. Methods & methods: Cholesterol was conjugated to chitosan (15 kDa) to form self-assembled micelles, which loaded curcumin. Physicochemical characterization and formulation optimization of the drug-loaded micelles (curcumin-loaded chitosan-cholesterol micelles [C-CCM]) were performed. In vitro cellular uptake and viability of C-CCM were investigated in melanoma and breast cancer cell lines. The antitumor efficacy was evaluated in 3D lung cancer spheroid model. The optimized C-CCM had size of approximately 162 nm with loading efficiency of approximately 36%. C-CCM was taken up efficiently by the cells, and it reduced cancer cell viability significantly compared with free curcumin. C-CCM enhanced the antitumor efficacy in spheroids, suggesting that C-CCM could be used as an effective chemotherapy in cancer.

  4. Structural Transition in a Fluid of Spheroids: A Low-Density Vestige of Jamming.

    Science.gov (United States)

    Cohen, A P; Dorosz, S; Schofield, A B; Schilling, T; Sloutskin, E

    2016-03-04

    A thermodynamically equilibrated fluid of hard spheroids is a simple model of liquid matter. In this model, the coupling between the rotational degrees of freedom of the constituent particles and their translations may be switched off by a continuous deformation of a spheroid of aspect ratio t into a sphere (t=1). We demonstrate, by experiments, theory, and computer simulations, that dramatic nonanalytic changes in structure and thermodynamics of the fluids take place, as the coupling between rotations and translations is made to vanish. This nonanalyticity, reminiscent of a second-order liquid-liquid phase transition, is not a trivial consequence of the shape of an individual particle. Rather, free volume considerations relate the observed transition to a similar nonanalyticity at t=1 in structural properties of jammed granular ellipsoids. This observation suggests a deep connection to exist between the physics of jamming and the thermodynamics of simple fluids.

  5. Miniaturized microscope for high throughput screening of tumor spheroids in microfluidic devices

    Science.gov (United States)

    Uranga, Javier; Rodríguez-Pena, Alejandro; Gahigiro, Desiré; Ortiz-de-Solorzano, Carlos

    2018-02-01

    High-throughput in vitro screening of highly physiological three-dimensional cell cultures (3D-HTS) is rapidly gaining importance in preclinical studies, to study the effect of the microenvironment in tumor development, and to evaluate the efficacy of new anticancer drugs. Furthermore, it could also be envisioned the use of 3D-HTS systems in personalized anti-cancer treatment planning, based on tumor organoids or spheroids grown from tumor biopsies or isolated tumor circulating cells. Most commercial, multi-well plate based 3D-HTS systems are large, expensive, and are based on the use of multi-well plates that hardly provide a physiological environment and require the use of large amounts of biological material and reagents. In this paper we present a novel, miniaturized inverted microscope (hereinafter miniscospe), made up of low-cost, mass producible parts, that can be used to monitor the growth of living tumor cell spheroids within customized three-dimensional microfluidic platforms. Our 3D-HTS miniscope combines phase contrast imaging based on oblique back illumination technique with traditional widefield epi-fluorescence imaging, implemented using miniaturized electro-optical parts and gradient-index refraction lenses. This small (3x6x2cm), lightweight device can effectively image overtime the growth of (>200) tumor spheroids in a controlled and reproducible environment. Our miniscope can be used to acquire time-lapse images of cellular living spheroids over the course of several hours and captures their growth before and after drug treatment, to evaluate the effectiveness of the drug.

  6. Chemical composition of extremely metal-poor stars in the Sextans dwarf spheroidal galaxy

    OpenAIRE

    Aoki, W.; Arimoto, N.; Sadakane, K.; Tolstoy, E.; Battaglia, G.; Jablonka, P.; Shetrone, M.; Letarte, B.; Irwin, M.; Hill, V.; Francois, P.; Venn, K.; Primas, F.; Helmi, A.; Kaufer, A.

    2009-01-01

    Context. Individual stars in dwarf spheroidal galaxies around the Milky Way Galaxy have been studied both photometrically and spectroscopically. Extremely metal-poor stars among them are very valuable because they should record the early enrichment in the Local Group. However, our understanding of these stars is very limited because detailed chemical abundance measurements are needed from high resolution spectroscopy. Aims. To constrain the formation and chemical evolution of dwarf galaxi...

  7. Hydrogen atom and the H+2 and HeH++ molecular ions inside prolate spheroidal boxes

    International Nuclear Information System (INIS)

    Ley-Koo, E.; Cruz, S.A.

    1981-01-01

    We formulate the exact solution of the Schroedinger equation for systems of one electron in the Coulomb field of one or two fixed nuclei at the foci inside prolate spheroidal boxes. Numerical results are obtained for the energy eigenvalues and eigenfunctions of the lowest states of the hydrogen atom and the H + 2 and HeH ++ molecular ions for boxes of different sizes and eccentricities. We also evaluate the hyperfine splitting of atomic hydrogen and of H + 2

  8. Multipoles and Force on External Points for a Two-layered Spheroidal Liquid Mass Rotating Differentialy

    Energy Technology Data Exchange (ETDEWEB)

    Cisneros-Parra, Joel U. [Facultad de Ciencias, UASLP, Zona Universitaria, San Luis Potosi, S,L,P, 78290 (Mexico); Martinez-Herrera, Francisco J.; Montalvo-Castro, J. Daniel [Instituto de Fisica, UASLP, Zona Universitaria, San Luis Potosi, S,L,P, 78290 (Mexico)

    2017-10-20

    We recently reported on a series of equilibrium figures for a self-gravitating heterogeneous liquid body, consisting of two concentric distorted spheroids, “nucleus” and “atmosphere,” each endowed with its own internal motion of differential rotation. In our current work, we calculate the body’s force at external points and obtain a multipolar expansion of the potential. We also give an account of figures with prolate nuclei, which remained unnoticed by us in our former paper.

  9. Model-independent constraints on dark matter annihilation in dwarf spheroidal galaxies

    OpenAIRE

    Boddy, Kimberly K.; Kumar, Jason; Marfatia, Danny; Sandick, Pearl

    2018-01-01

    We present a general, model-independent formalism for determining bounds on the production of photons in dwarf spheroidal galaxies via dark matter annihilation, applicable to any set of assumptions about dark matter particle physics or astrophysics. As an illustration, we analyze gamma-ray data from the Fermi Large Area Telescope to constrain a variety of nonstandard dark matter models, several of which have not previously been studied in the context of dwarf galaxy searches.

  10. Wearing Quality of Austenitic, Duplex Cast Steel, Gray and Spheroidal Graphite Iron

    Directory of Open Access Journals (Sweden)

    S. Pietrowski

    2012-04-01

    Full Text Available The current work presents the research results of abrasion wear and adhesive wear at rubbing and liquid friction of new austenitic, austenitic-ferritic (“duplex” cast steel and gray cast iron EN-GJL-250, spheroidal graphite iron EN-GJS-600-3, pearlitic with ledeburitic carbides and spheroidal graphite iron with ledeburitic carbides with a microstructure of the metal matrix: pearlitic, upper bainite, mixture of upper and lower bainite, martensitic with austenite, pearlitic-martensitic-bainitic-ausferritic obtained in the raw state. The wearing quality test was carried out on a specially designed and made bench. Resistance to abrasion wear was tested using sand paper P40. Resistance to adhesive wear was tested in interaction with steel C55 normalized, hardened and sulfonitrided. The liquid friction was obtained using CASTROL oil. It was stated that austenitic cast steel and “duplex” are characterized by a similar value of abrasion wear and adhesive wear at rubbing friction. The smallest decrease in mass was shown by the cast steel in interaction with the sulfonitrided steel C55. Austenitic cast steel and “duplex��� in different combinations of friction pairs have a higher wear quality than gray cast iron EN-GJL- 250 and spheroidal graphite iron EN-GJS-600-3. Austenitic cast steel and “duplex” are characterized by a lower wearing quality than the spheroidal graphite iron with bainitic-martensitic microstructure. In the adhesive wear test using CASTROL oil the tested cast steels and cast irons showed a small mass decrease within the range of 1÷2 mg.

  11. Spheroidal and conical shapes of ferrofluid-filled capsules in magnetic fields

    Science.gov (United States)

    Wischnewski, Christian; Kierfeld, Jan

    2018-04-01

    We investigate the deformation of soft spherical elastic capsules filled with a ferrofluid in external uniform magnetic fields at fixed volume by a combination of numerical and analytical approaches. We develop a numerical iterative solution strategy based on nonlinear elastic shape equations to calculate the stretched capsule shape numerically and a coupled finite element and boundary element method to solve the corresponding magnetostatic problem and employ analytical linear response theory, approximative energy minimization, and slender-body theory. The observed deformation behavior is qualitatively similar to the deformation of ferrofluid droplets in uniform magnetic fields. Homogeneous magnetic fields elongate the capsule and a discontinuous shape transition from a spheroidal shape to a conical shape takes place at a critical field strength. We investigate how capsule elasticity modifies this hysteretic shape transition. We show that conical capsule shapes are possible but involve diverging stretch factors at the tips, which gives rise to rupture for real capsule materials. In a slender-body approximation we find that the critical susceptibility above which conical shapes occur for ferrofluid capsules is the same as for droplets. At small fields capsules remain spheroidal and we characterize the deformation of spheroidal capsules both analytically and numerically. Finally, we determine whether wrinkling of a spheroidal capsule occurs during elongation in a magnetic field and how it modifies the stretching behavior. We find the nontrivial dependence between the extent of the wrinkled region and capsule elongation. Our results can be helpful in quantitatively determining capsule or ferrofluid material properties from magnetic deformation experiments. All results also apply to elastic capsules filled with a dielectric liquid in an external uniform electric field.

  12. Axisymmetric force-free states and relaxation of a spheroidal spheromak

    International Nuclear Information System (INIS)

    Throumoulopoulos, G.N.; Pantis, G.

    1990-01-01

    Axisymmetric force-free equilibrium eigenstates for a prolate as well as an oblate spheroidal Spheromak with arbitrary elongation are obtained. In the framework of the Woltjer-Taylor relaxation theory the relaxed states are also identified. A simple hypothesis for the relaxation process is introduced, which implies that the plasma relaxes from multitoroidal formations to a singly toroidal configuration, in qualitative agreement with experimental results. (author)

  13. Theory of axially symmetric probes in a collisionless magnetoplasma: Aligned spheroids, finite cylinders, and disks

    International Nuclear Information System (INIS)

    Rubinstein, J.; Laframboise, J.G.

    1983-01-01

    A theory is presented for current collection by electrostatic probes in a collisionless, Maxwellian plasma containing a uniform magnetic field B, where the probes are spheroids or finite cylinders whose axis of symmetry is aligned with B, or disks perpendicular to B. The theory yields upper-bound and adiabatic-limit currents for the attracted particle species. For the repelled species, it yields upper and lower bounds. This work is an extension of existing theory for spherical probes by Rubinstein and Laframboise

  14. Numerical Simulation of Spheroidization Process of TiAl Alloy Powders in Radio Frequency Plasma

    OpenAIRE

    ZHU Langping; LU Xin; LIU Chengcheng; LI Jianchong; NAN Hai

    2017-01-01

    A numerical simulation method was used to study the radio frequency plasma spheroidization process of TiAl alloy powder. The effects of velocity field and temperature field on the motion trajectory and mass change of TiAl alloy powder with different particle size were analyzed.The results show that the temperature of powder particles increases rapidly under high temperature plasma, surface evaporation cause the reduction of particle size, and particles with small size tend to evaporate quickl...

  15. Multipoles and Force on External Points for a Two-layered Spheroidal Liquid Mass Rotating Differentialy

    International Nuclear Information System (INIS)

    Cisneros-Parra, Joel U.; Martinez-Herrera, Francisco J.; Montalvo-Castro, J. Daniel

    2017-01-01

    We recently reported on a series of equilibrium figures for a self-gravitating heterogeneous liquid body, consisting of two concentric distorted spheroids, “nucleus” and “atmosphere,” each endowed with its own internal motion of differential rotation. In our current work, we calculate the body’s force at external points and obtain a multipolar expansion of the potential. We also give an account of figures with prolate nuclei, which remained unnoticed by us in our former paper.

  16. Hydrolysis of phosphatidylcholine by hepatic lipase in discoidal and spheroidal recombinant high-density lipoprotein.

    Science.gov (United States)

    Tansey, J T; Thuren, T Y; Jerome, W G; Hantgan, R R; Grant, K; Waite, M

    1997-10-07

    Hepatic lipase (HL) hydrolysis of phosphatidylcholine (PC) was studied in recombinant high-density lipoprotein particles (r-HDL). r-HDL were made from cholate mixed micelles that contained PC, apo AI, and, in some cases, unesterified cholesterol. r-HDL were characterized using chemical composition, nondenaturing gradient gel electrophoresis, transmission electron microscopy, and dynamic light scattering. The r-HDL were found to be discoidal and in the size range of native HDL. Upon treatment of cholesterol-containing r-HDL with lecithin-cholesterol acyltransferase (LCAT), to form cholesteryl ester, the discoidal r-HDL became spheroidal. The effects of r-HDL morphology and size on HL activity were studied on r-HDL made of palmitoyloleoyl-PC, unesterified cholesterol, cholesteryl ester, and apolipoprotein AI. Spheroidal r-HDL were hydrolyzed at a faster rate than discoidal r-HDL. Protein-poor r-HDL were hydrolyzed by HL at a faster rate than protein rich r-HDL. Unesterified cholesterol had no apparent effect on particle PC hydrolysis. The hydrolysis of different species of PC [dipalmitoyl (DPPC), dioleoyl(DOPC), palmitoylarachidonoyl (PAPC), and palmitoyloleoyl (POPC)] in r-HDL was also investigated. In discoidal r-HDL, we found that POPC >/= DOPC = PAPC/DPPC. However, in LCAT-treated spheroidal r-HDL, POPC = DOPC > PAPC/DPPC. In both discoidal and spheroidal rHDL, DPPC containing r-HDL were not hydrolyzed to a significant extent. Collectively, these studies demonstrate that the physico-chemical properties of particles (such as phospholipid packing and phospholipid acyl composition) play a significant role in hydrolysis of HDL phospholipid by HL and, therefore, in reverse cholesterol transport.

  17. Spheroidal degeneration in H626R TGFBI variant lattice dystrophy: a multimodality analysis.

    Science.gov (United States)

    Lai, Kevin; Reidy, Jason; Bert, Benjamin; Milman, Tatyana

    2014-07-01

    The aim of this study was to describe clinical, imaging, molecular genetic, histopathologic, immunohistochemical, and ultrastructural characteristics of coexistent amyloid and spheroidal degeneration-type deposits in a family with histidine-626-arginine transforming growth factor beta-induced (H626R TGFBI) variant lattice dystrophy. This is a retrospective clinical-pathological and genetic analysis of one family with H626R variant lattice dystrophy. Pedigree analysis showed an autosomal dominant inheritance pattern of the disease. Examination of 3 affected family members revealed asymmetric, thick, branching lattice-like deposits associated with corneal haze. Sequencing of the TGFBI gene revealed a high-penetrance disease-causing sequence variation (H626R CAT>CGT heterozygous). Optical coherence tomography demonstrated fusiform, poorly demarcated hyperechoic stromal deposits with focal hypoechoic central regions. Histology of the corneal discs from 2 affected family members showed stromal deposits consistent with TGFBI amyloid. Some amyloid deposits contained a central nidus of spheroidal degeneration-type material that demonstrated autofluorescence, stained with elastic and Masson trichrome stains, did not stain with periodic acid-Schiff or Congo red stains, was nonbirefringent, and did not immunoreact with keratoepithelin antibodies. Transmission electron microscopy confirmed the presence of amyloid fibrils with central, electrodense, homogeneous, discrete, spheroidal degeneration-type deposits. The presence of spheroidal deposits in a subset of affected patients, variability in presentation within an individual and between family members, predominant anterior corneal stromal location and nonimmunoreactivity of deposits for keratoepithelin suggest that these deposits are degenerative in nature. The deposits may arise from ultraviolet light-altered proteins diffused from the limbus, which form a nidus for keratoepithelin deposition.

  18. Axisymmetric force-free states and relaxation of a spheroidal spheromak

    International Nuclear Information System (INIS)

    Throumoulopoulos, G.N.; Pantis, G.

    1990-01-01

    Axisymmetric force-free equilibrium eigenstates for a prolate as well as an oblate spheroidal spheromak with arbitrary elongation are obtained. In the framework of the Woltjer-Taylor relaxation theory the relaxed states are also identified. A simple hypothesis for the relaxation process is introduced which implies that the plasma relaxes from multitoroidal formations to a singly toroidal configuration in qualitative agreement with experimental results. (Author)

  19. Numerical simulation of water exit of an initially fully submerged buoyant spheroid in an axisymmetric flow

    Energy Technology Data Exchange (ETDEWEB)

    Ni, B Y; Wu, G X, E-mail: g.wu@ucl.ac.uk [College of Shipbuilding Engineering, Harbin Engineering University, Harbin 150001 (China)

    2017-08-15

    The free water exit of an initially fully submerged buoyant spheroid in an axisymmetric flow, which is driven by the difference between the vertical fluid force and gravity, is investigated. The fluid is assumed to be incompressible and inviscid, and the flow to be irrotational. The velocity potential theory is adopted together with fully nonlinear boundary conditions on the free surface. The surface tension is neglected and the pressure is taken as constant on the free surface. The acceleration of the body at each time step is obtained as part of the solution. Its nonlinear mutual dependence on the fluid force is decoupled through the auxiliary function method. The free-surface breakup by body penetration and water detachment from the body are treated through numerical conditions. The slender body theory based on the zero potential assumption on the undisturbed flat free surface is adopted, through which a condition for full water exit of a spheroid is obtained. Comparison is made between the results from the slender body theory and from the fully nonlinear theory through the boundary-element method, and good agreement is found when the spheroid is slender. Extensive case studies are undertaken to investigate the effects of body density, dimensions and the initial submergence. (paper)

  20. Characterization and reproducibility of HepG2 hanging drop spheroids toxicology in vitro.

    Science.gov (United States)

    Hurrell, Tracey; Ellero, Andrea Antonio; Masso, Zelie Flavienne; Cromarty, Allan Duncan

    2018-02-21

    Hepatotoxicity remains a major challenge in drug development despite preclinical toxicity screening using hepatocytes of human origin. To overcome some limitations of reproducing the hepatic phenotype, more structurally and functionally authentic cultures in vitro can be introduced by growing cells in 3D spheroid cultures. Characterisation and reproducibility of HepG2 spheroid cultures using a high-throughput hanging drop technique was performed and features contributing to potential phenotypic variation highlighted. Cultured HepG2 cells were seeded into Perfecta 3D® 96-well hanging drop plates and assessed over time for morphology, viability, cell cycle distribution, protein content and protein-mass profiles. Divergent aspects which were assessed included cell stocks, seeding density, volume of culture medium and use of extracellular matrix additives. Hanging drops are advantageous due to no complex culture matrix being present, enabling background free extractions for downstream experimentation. Varying characteristics were observed across cell stocks and batches, seeding density, culture medium volume and extracellular matrix when using immortalized HepG2 cells. These factors contribute to wide-ranging cellular responses and highlights concerns with respect to generating a reproducible phenotype in HepG2 hanging drop spheroids. Copyright © 2018 Elsevier Ltd. All rights reserved.

  1. Hyperspherical Coulomb spheroidal basis in the Coulomb three-body problem

    International Nuclear Information System (INIS)

    Abramov, D. I.

    2013-01-01

    A hyperspherical Coulomb spheroidal (HSCS) representation is proposed for the Coulomb three-body problem. This is a new expansion in the set of well-known Coulomb spheroidal functions. The orthogonality of Coulomb spheroidal functions on a constant-hyperradius surface ρ = const rather than on a constant-internuclear-distance surface R = const, as in the traditional Born-Oppenheimer approach, is a distinguishing feature of the proposed approach. Owing to this, the HSCS representation proves to be consistent with the asymptotic conditions for the scattering problem at energies below the threshold for three-body breakup: only a finite number of radial functions do not vanish in the limit of ρ→∞, with the result that the formulation of the scattering problem becomes substantially simpler. In the proposed approach, the HSCS basis functions are considerably simpler than those in the well-known adiabatic hyperspherical representation, which is also consistent with the asymptotic conditions. Specifically, the HSCS basis functions are completely factorized. Therefore, there arise no problems associated with avoided crossings of adiabatic hyperspherical terms.

  2. Dark Matter Searches with the Fermi-LAT in the Direction of Dwarf Spheroidals

    Energy Technology Data Exchange (ETDEWEB)

    Wood, Matthew; Anderson, Brandon; Drlica-Wagner, Alex; Cohen-Tanugi, Johann; Conrad, Jan

    2015-07-13

    The dwarf spheroidal satellite galaxies of the Milky Way are some of the most dark-matter-dominated objects known. Due to their proximity, high dark matter content, and lack of astrophysical backgrounds, dwarf spheroidal galaxies are widely considered to be among the most promising targets for the indirect detection of dark matter via gamma rays. Here we report on gamma-ray observations of Milky Way dwarf spheroidal satellite galaxies based on 6 years of Fermi Large Area Telescope data processed with the new Pass 8 reconstruction and event-level analysis. None of the dwarf galaxies are significantly detected in gamma rays, and we present upper limits on the dark matter annihilation cross section from a combined analysis of the 15 most promising dwarf galaxies. The constraints derived are among the strongest to date using gamma rays, and lie below the canonical thermal relic cross section for WIMPs of mass ≲ 100GeV annihilating via the bb-bar and τ⁺τ⁻ channels.

  3. Classification of materials for conducting spheroids based on the first order polarization tensor

    Science.gov (United States)

    Khairuddin, TK Ahmad; Mohamad Yunos, N.; Aziz, ZA; Ahmad, T.; Lionheart, WRB

    2017-09-01

    Polarization tensor is an old terminology in mathematics and physics with many recent industrial applications including medical imaging, nondestructive testing and metal detection. In these applications, it is theoretically formulated based on the mathematical modelling either in electrics, electromagnetics or both. Generally, polarization tensor represents the perturbation in the electric or electromagnetic fields due to the presence of conducting objects and hence, it also desribes the objects. Understanding the properties of the polarization tensor is necessary and important in order to apply it. Therefore, in this study, when the conducting object is a spheroid, we show that the polarization tensor is positive-definite if and only if the conductivity of the object is greater than one. In contrast, we also prove that the polarization tensor is negative-definite if and only if the conductivity of the object is between zero and one. These features categorize the conductivity of the spheroid based on in its polarization tensor and can then help to classify the material of the spheroid.

  4. Thermal Plasma Spheroidization of High-Nitrogen Stainless Steel Powder Alloys Synthesized by Mechanical Alloying

    Science.gov (United States)

    Razumov, Nikolay G.; Popovich, Anatoly A.; Wang, QingSheng

    2018-03-01

    This paper presents the results of experimental studies on the treatment of Fe-23Cr-11Mn-1N high-nitrogen stainless steel powder alloys, synthesized by the mechanical alloying (MA) of elemental powders in the flow of a thermal plasma. Fe-23Cr-11Mn-1N high-nitrogen stainless steel powder alloys were prepared by MA in the attritor under an argon atmosphere. For spheroidization of Fe-23Cr-11Mn-1N high-nitrogen stainless steel powder alloys, the TekSphero 15 plant manufactured by Tekna Plasma Systems Inc was used. The studies have shown the possibility of obtaining Fe-23Cr-11Mn-1N high-nitrogen spherical powders steel alloys from the powder obtained by MA. According to the results of a series of experiments, it was found that the results of plasma spheroidization of powders essentially depend on the size of the fraction due to some difference in the particle shape and flowability, and on the gas regime of the plasma torch. It is established that during the plasma spheroidization process, some of the nitrogen leaves the alloy. The loss rate of nitrogen depends on the size of the initial particles.

  5. Design of a uranium-dioxide powder spheroidization system by plasma processing

    Science.gov (United States)

    Cavender, Daniel

    The plasma spheroidization system (PSS) is the first process in the development of a tungsten-uranium dioxide (W-UO2) ceramic-metallic (cermet) fuel for nuclear thermal rocket (NTR) propulsion. For the purposes of fissile fuel retention, UO2 spheroids ranging in size from 50 - 100 micrometers (μm) in diameter will be encapsulated in a tungsten shell. The PSS produces spherical particles by melting angular stock particles in an argon-hydrogen plasma jet where they become spherical due to surface tension. Surrogate CeO 2 powder was used in place of UO2 for system and process parameter development. Stock and spheroidized powders were micrographed using optical and scanning electron microscopy and evaluated by statistical methods to characterize and compare the spherocity of pre and post process powders. Particle spherocity was determined by irregularity parameter. Processed powders showed a statistically significant improvement in spherocity, with greater that 60% of the examined particles having an irregularity parameter of equal to or lower than 1.2, compared to stock powder.

  6. Two-dimensional particle-in-cell plasma source ion implantation of a prolate spheroid target

    International Nuclear Information System (INIS)

    Cheng-Sen, Liu; Hong-Ying, Han; Xiao-Qing, Peng; Ye, Chang; De-Zhen, Wang

    2010-01-01

    A two-dimensional particle-in-cell simulation is used to study the time-dependent evolution of the sheath surrounding a prolate spheroid target during a high voltage pulse in plasma source ion implantation. Our study shows that the potential contour lines pack more closely in the plasma sheath near the vertex of the major axis, i.e. where a thinner sheath is formed, and a non-uniform total ion dose distribution is incident along the surface of the prolate spheroid target due to the focusing of ions by the potential structure. Ion focusing takes place not only at the vertex of the major axis, where dense potential contour lines exist, but also at the vertex of the minor axis, where sparse contour lines exist. This results in two peaks of the received ion dose, locating at the vertices of the major and minor axes of the prolate spheroid target, and an ion dose valley, staying always between the vertices, rather than at the vertex of the minor axis

  7. Two-dimensional particle-in-cell plasma source ion implantation of a prolate spheroid target

    Science.gov (United States)

    Liu, Cheng-Sen; Han, Hong-Ying; Peng, Xiao-Qing; Chang, Ye; Wang, De-Zhen

    2010-03-01

    A two-dimensional particle-in-cell simulation is used to study the time-dependent evolution of the sheath surrounding a prolate spheroid target during a high voltage pulse in plasma source ion implantation. Our study shows that the potential contour lines pack more closely in the plasma sheath near the vertex of the major axis, i.e. where a thinner sheath is formed, and a non-uniform total ion dose distribution is incident along the surface of the prolate spheroid target due to the focusing of ions by the potential structure. Ion focusing takes place not only at the vertex of the major axis, where dense potential contour lines exist, but also at the vertex of the minor axis, where sparse contour lines exist. This results in two peaks of the received ion dose, locating at the vertices of the major and minor axes of the prolate spheroid target, and an ion dose valley, staying always between the vertices, rather than at the vertex of the minor axis.

  8. Study of the boxlike dark matter signals from dwarf spheroidal galaxies with Fermi-LAT data

    Science.gov (United States)

    Li, Shang; Liang, Yun-Feng; Xia, Zi-Qing; Zu, Lei; Duan, Kai-Kai; Shen, Zhao-Qiang; Feng, Lei; Yuan, Qiang; Fan, Yi-Zhong

    2018-04-01

    The observation of a special spectral feature in the gamma-ray data would be one of the best ways to identify dark matter (DM). The box-shaped gamma-ray spectra could be generated by the decay of intermediate particles produced by DM annihilation or decay. It provides another kind of signal that can be relatively easily distinguished from astrophysical backgrounds besides the linelike signals. Dwarf spheroidal galaxies are expected to be dominated by DM and may be one of the most promising targets for indirect DM searches. In this paper, we study the box-shaped DM signals with Fermi-LAT observations of dwarf spheroidal galaxies. We analyze 106 months of Fermi-LAT data to derive the upper limits on the annihilation cross section or the decay timescale of DM. In addition, we compare the results for different sample selections and DM density distributions. We expect that more dwarf spheroidal galaxies will be found and the sensitivity of box-shaped gamma-ray signal searches will be significantly improved in the future.

  9. Human adipose-derived stem cell spheroid treated with photobiomodulation irradiation accelerates tissue regeneration in mouse model of skin flap ischemia.

    Science.gov (United States)

    Park, In-Su; Chung, Phil-Sang; Ahn, Jin Chul; Leproux, Anais

    2017-11-01

    Skin flap grafting is a form of transplantation widely used in plastic surgery. However, ischemia/reperfusion injury is the main factor which reduces the survival rate of flaps following grafting. We investigated whether photobiomodulation (PBM) precondition prior to human adipose-derived stromal cell (hASC) spheroid (PBM-spheroid) transplantation improved skin tissue functional recovery by the stimulation of angiogenesis and tissue regeneration in skin flap of mice. The LED had an emission wavelength peaked at 660 ± 20 nm (6 J/cm 2 , 10 mW/cm 2 ). The expression of angiogenic growth factors in PBM-spheroid hASCs was much greater than that of not-PBM-treated spheroid or monolayer-cultured hASCs. From immunochemical staining analysis, the hASCs of PBM-spheroid were CD31 + , KDR + , and CD34 + , whereas monolayer-cultured hASCs were negative for these markers. To evaluate the therapeutic effect of hASC PBM-spheroid in vivo, PBS, monolayer-cultured hASCs, and not-PBM-spheroid were transplanted into a skin flap model. The animals were observed for 14 days. The PBM-spheroid hASCs transplanted into the skin flap ischemia differentiated into endothelial cells and remained differentiated. Transplantation of PBM-spheroid hASCs into the skin flap ischemia significantly elevated the density of vascular formations through angiogenic factors released by the skin flap ischemia and enhanced tissue regeneration at the lesion site. Consistent with these results, the transplantation of PBM-spheroid hASCs significantly improved functional recovery compared with PBS, monolayer-cultured hASCs, and not-PBM-spheroid treatment. These findings suggest that transplantation of PBM-spheroid hASCs may be an effective stem cell therapy for the treatment of skin flap ischemia.

  10. Respiratory hazard assessment of combined exposure to complete gasoline exhaust and respirable volcanic ash in a multicellular human lung model at the air-liquid interface

    Science.gov (United States)

    Tomasek, Ines; Horwell, Claire J.; Bisig, Christoph; Damby, David; Comte, Pierre; Czerwinski, Jan; Petri-Fink, Alke; Clift, Martin J D; Drasler, Barbara; Rothen-Rutishauer, Barbara

    2018-01-01

    Communities resident in urban areas located near active volcanoes can experience volcanic ash exposures during, and following, an eruption, in addition to sustained exposures to high concentrations of anthropogenic air pollutants (e.g., vehicle exhaust emissions). Inhalation of anthropogenic pollution is known to cause the onset of, or exacerbate, respiratory and cardiovascular diseases. It is further postulated similar exposure to volcanic ash can also affect such disease states. Understanding of the impact of combined exposure of volcanic ash and anthropogenic pollution to human health, however, remains limited.The aim of this study was to assess the biological impact of combined exposure to respirable volcanic ash (from Soufrière Hills volcano (SHV), Montserrat and Chaitén volcano (ChV), Chile; representing different magmatic compositions and eruption styles) and freshly-generated complete exhaust from a gasoline vehicle. A multicellular human lung model (an epithelial cell-layer composed of A549 alveolar type II-like cells complemented with human blood monocyte-derived macrophages and dendritic cells cultured at the air-liquid interface) was exposed to diluted exhaust (1:10) continuously for 6 h, followed by immediate exposure to the ash as a dry powder (0.54 ± 0.19 μg/cm2 and 0.39 ± 0.09 μg/cm2 for SHV and ChV ash, respectively). After an 18 h incubation, cells were exposed again for 6 h to diluted exhaust, and a final 18 h incubation (at 37 °C and 5% CO2). Cell cultures were then assessed for cytotoxic, oxidative stress and (pro-)inflammatory responses.Results indicate that, at all tested (sub-lethal) concentrations, co-exposures with both ash samples induced no significant expression of genes associated with oxidative stress (HMOX1, NQO1) or production of (pro-)inflammatory markers (IL-1β, IL-8, TNF-α) at the gene and protein levels. In summary, considering the employed experimental conditions, combined exposure of

  11. Respiratory hazard assessment of combined exposure to complete gasoline exhaust and respirable volcanic ash in a multicellular human lung model at the air-liquid interface.

    Science.gov (United States)

    Tomašek, Ines; Horwell, Claire J; Bisig, Christoph; Damby, David E; Comte, Pierre; Czerwinski, Jan; Petri-Fink, Alke; Clift, Martin J D; Drasler, Barbara; Rothen-Rutishauser, Barbara

    2018-07-01

    Communities resident in urban areas located near active volcanoes can experience volcanic ash exposures during, and following, an eruption, in addition to sustained exposures to high concentrations of anthropogenic air pollutants (e.g., vehicle exhaust emissions). Inhalation of anthropogenic pollution is known to cause the onset of, or exacerbate, respiratory and cardiovascular diseases. It is further postulated similar exposure to volcanic ash can also affect such disease states. Understanding of the impact of combined exposure of volcanic ash and anthropogenic pollution to human health, however, remains limited. The aim of this study was to assess the biological impact of combined exposure to respirable volcanic ash (from Soufrière Hills volcano (SHV), Montserrat and Chaitén volcano (ChV), Chile; representing different magmatic compositions and eruption styles) and freshly-generated complete exhaust from a gasoline vehicle. A multicellular human lung model (an epithelial cell-layer composed of A549 alveolar type II-like cells complemented with human blood monocyte-derived macrophages and dendritic cells cultured at the air-liquid interface) was exposed to diluted exhaust (1:10) continuously for 6 h, followed by immediate exposure to the ash as a dry powder (0.54 ± 0.19 μg/cm 2 and 0.39 ± 0.09 μg/cm 2 for SHV and ChV ash, respectively). After an 18 h incubation, cells were exposed again for 6 h to diluted exhaust, and a final 18 h incubation (at 37 °C and 5% CO 2 ). Cell cultures were then assessed for cytotoxic, oxidative stress and (pro-)inflammatory responses. Results indicate that, at all tested (sub-lethal) concentrations, co-exposures with both ash samples induced no significant expression of genes associated with oxidative stress (HMOX1, NQO1) or production of (pro-)inflammatory markers (IL-1β, IL-8, TNF-α) at the gene and protein levels. In summary, considering the employed experimental conditions, combined exposure of

  12. The diversification of the LIM superclass at the base of the metazoa increased subcellular complexity and promoted multicellular specialization.

    Directory of Open Access Journals (Sweden)

    Bernard J Koch

    Full Text Available Throughout evolution, the LIM domain has been deployed in many different domain configurations, which has led to the formation of a large and distinct group of proteins. LIM proteins are involved in relaying stimuli received at the cell surface to the nucleus in order to regulate cell structure, motility, and division. Despite their fundamental roles in cellular processes and human disease, little is known about the evolution of the LIM superclass.We have identified and characterized all known LIM domain-containing proteins in six metazoans and three non-metazoans. In addition, we performed a phylogenetic analysis on all LIM domains and, in the process, have identified a number of novel non-LIM domains and motifs in each of these proteins. Based on these results, we have formalized a classification system for LIM proteins, provided reasonable timing for class and family origin events; and identified lineage-specific loss events. Our analysis is the first detailed description of the full set of LIM proteins from the non-bilaterian species examined in this study.Six of the 14 LIM classes originated in the stem lineage of the Metazoa. The expansion of the LIM superclass at the base of the Metazoa undoubtedly contributed to the increase in subcellular complexity required for the transition from a unicellular to multicellular lifestyle and, as such, was a critically important event in the history of animal multicellularity.

  13. The phosphatomes of the multicellular myxobacteria Myxococcus xanthus and Sorangium cellulosum in comparison with other prokaryotic genomes.

    Directory of Open Access Journals (Sweden)

    Anke Treuner-Lange

    Full Text Available BACKGROUND: Analysis of the complete genomes from the multicellular myxobacteria Myxococcus xanthus and Sorangium cellulosum identified the highest number of eukaryotic-like protein kinases (ELKs compared to all other genomes analyzed. High numbers of protein phosphatases (PPs could therefore be anticipated, as reversible protein phosphorylation is a major regulation mechanism of fundamental biological processes. METHODOLOGY: Here we report an intensive analysis of the phosphatomes of M. xanthus and S. cellulosum in which we constructed phylogenetic trees to position these sequences relative to PPs from other prokaryotic organisms. PRINCIPAL FINDINGS: PREDOMINANT OBSERVATIONS WERE: (i M. xanthus and S. cellulosum possess predominantly Ser/Thr PPs; (ii S. cellulosum encodes the highest number of PP2c-type phosphatases so far reported for a prokaryotic organism; (iii in contrast to M. xanthus only S. cellulosum encodes high numbers of SpoIIE-like PPs; (iv there is a significant lack of synteny among M. xanthus and S. cellulosum, and (v the degree of co-organization between kinase and phosphatase genes is extremely low in these myxobacterial genomes. CONCLUSIONS: We conclude that there has been a greater expansion of ELKs than PPs in multicellular myxobacteria.

  14. Fire exposed aluminium structures

    NARCIS (Netherlands)

    Maljaars, J.; Fellinger, J.E.J.; Soetens, F.

    2005-01-01

    Material properties and mechanical response models for fire design of steel structures are based on extensive research and experience. Contrarily, the behaviour of aluminium load bearing structures exposed to fire is relatively unexplored. This article gives an overview of physical