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Sample records for cell surface combined

  1. Detection of radiolabelled bone marrow cells bearing surface immunoglobulins by combined autoradiography and immunoperoxidase

    International Nuclear Information System (INIS)

    A method is described for the simultaneous detection of radiolabelled bone marrow cells bearing surface immunoglobulins by combined autoradiography and immunoperoxidase. Bone marrow cells from normal CBA mice prelabelled in vivo with 125IUDR or exposed in vitro to [3H]thymidine were incubated with rabbit anti-mouse immunoglobulins under capping conditions, washed, cytocentrifuged and treated with methanol and hydrogen peroxide to destroy endogenous peroxidase. Cells were then covered with peroxidase-conjugated goat anti-rabbit immunoglobulins, washed, treated with diaminobenzidine and hydrogen peroxide and finally covered with autoradiographic stripping film and exposed for different times. Peroxidase-positive cells were typically capped and those radiolabelled had autoradiographic silver grains overlying the nucleus. (Auth.)

  2. Tumor suppressor protein SMAR1 modulates the roughness of cell surface: combined AFM and SEM study

    Directory of Open Access Journals (Sweden)

    Mamgain Hitesh

    2009-10-01

    Full Text Available Abstract Background Imaging tools such as scanning electron microscope (SEM and atomic force microscope (AFM can be used to produce high-resolution topographic images of biomedical specimens and hence are well suited for imaging alterations in cell morphology. We have studied the correlation of SMAR1 expression with cell surface smoothness in cell lines as well as in different grades of human breast cancer and mouse tumor sections. Methods We validated knockdown and overexpression of SMAR1 using RT-PCR as well as Western blotting in human embryonic kidney (HEK 293, human breast cancer (MCF-7 and mouse melanoma (B16F1 cell lines. The samples were then processed for cell surface roughness studies using atomic force microscopy (AFM and scanning electron microscopy (SEM. The same samples were used for microarray analysis as well. Tumors sections from control and SMAR1 treated mice as well as tissues sections from different grades of human breast cancer on poly L-lysine coated slides were used for AFM and SEM studies. Results Tumor sections from mice injected with melanoma cells showed pronounced surface roughness. In contrast, tumor sections obtained from nude mice that were first injected with melanoma cells followed by repeated injections of SMAR1-P44 peptide, exhibited relatively smoother surface profile. Interestingly, human breast cancer tissue sections that showed reduced SMAR1 expression exhibited increased surface roughness compared to the adjacent normal breast tissue. Our AFM data establishes that treatment of cells with SMAR1-P44 results into increase in cytoskeletal volume that is supported by comparative gene expression data showing an increase in the expression of specific cytoskeletal proteins compared to the control cells. Altogether, these findings indicate that tumor suppressor function of SMAR1 might be exhibited through smoothening of cell surface by regulating expression of cell surface proteins. Conclusion Tumor suppressor

  3. Bicistronic retroviral vectors for combining myeloprotection with cell-surface marking.

    Science.gov (United States)

    Hildinger, M; Schilz, A; Eckert, H G; Bohn, W; Fehse, B; Zander, A; Ostertag, W; Baum, C

    1999-07-01

    We have developed a retroviral vector coexpressing the multidrug-resistance 1 (MDR1) cDNA for inducing cancer drug resistance and the truncated version of the low-affinity nerve growth factor receptor (DeltaLNGFR) for cell-surface marking of transduced cells. The vector is based on the FMEV backbone which mediates high levels of gene expression in hematopoietic cells. To achieve optimal expression levels of both cDNAs, untranslated regions from MDR1 and DeltaLNGFR were removed and three different connections were tested: retroviral splice signals, an internal ribosomal entry site (IRES) from encephalomyocarditis virus, and an internal promoter from the chicken beta-actin gene. As determined by two-color flow cytometry, the best correlation of the expression of both cDNAs was obtained using the vector SF1mSdelta which utilized retroviral splice signals for co-expression. Simultaneous expression of both cDNAs at the single cell level was also shown by confocal laser microscopy. Lymphoid and hematopoietic progenitor cells, including primary human CD34+ cells, transduced with SF1mSdelta acquired dominant multidrug resistance. Transduced primary CD34+ cells could be enriched in vitro based on expression of DeltaLNGFR, avoiding exposure to cytostatic agents. Thus, monitoring the selection of chemotherapy-resistant cells and analyzing their biological properties may be alleviated, both in vitro and in vivo. PMID:10455430

  4. A novel strategy for enrichment and isolation of osteoprogenitor cells from induced pluripotent stem cells based on surface marker combination.

    Directory of Open Access Journals (Sweden)

    Hiromi Ochiai-Shino

    Full Text Available In this study, we developed a new method to stimulate osteogenic differentiation in tissue-nonspecific alkaline phosphatase (TNAP-positive cells liberated from human induced pluripotent stem cells (hiPSCs-derived embryoid bodies (EBs with 14 days long TGF-β/IGF-1/FGF-2 treatment. TNAP is a marker protein of osteolineage cells. We analyzed and isolated TNAP-positive and E-cadherin-negative nonepithelial cells by fluorescence-activated cell sorting. Treating the cells with a combination of transforming growth factor (TGF-β, insulin-like growth factor (IGF-1, and fibroblast growth factor (FGF-2 for 14 days greatly enhanced TNAP expression and maximized expression frequency up to 77.3%. The isolated cells expressed high levels of osterix, which is an exclusive osteogenic marker. Culturing these TNAP-positive cells in osteoblast differentiation medium (OBM led to the expression of runt-related transcription factor 2, type I collagen, bone sialoprotein, and osteocalcin (OCN. These cells responded to treatment with activated vitamin D3 by upregulating OCN. Furthermore, in OBM they were capable of generating many mineralized nodules with strong expression of receptor activator of NF-kappaB ligand and sclerostin (SOST. Real-time RT-PCR showed a significant increase in the expression of osteocyte marker genes, including SOST, neuropeptide Y, and reelin. Scanning electron microscopy showed dendritic morphology. Examination of semi-thin toluidine blue-stained sections showed many interconnected dendrites. Thus, TNAP-positive cells cultured in OBM may eventually become terminally differentiated osteocyte-like cells. In conclusion, treating hiPSCs-derived cells with a combination of TGF-β, IGF-1, and FGF-2 generated TNAP-positive cells at high frequency. These TNAP-positive cells had a high osteogenic potential and could terminally differentiate into osteocyte-like cells. The method described here may reveal new pathways of osteogenesis and provide a novel

  5. Controlling cell growth on titanium by surface functionalization of heptylamine using a novel combined plasma polymerization mode.

    Science.gov (United States)

    Zhao, Jing H; Michalski, Wojtek P; Williams, Catherine; Li, Li; Xu, Hong-Sheng; Lamb, Peter R; Jones, Scott; Zhou, Yan M; Dai, Xiujuan J

    2011-05-01

    A novel bio-interface, produced by a combined plasma polymerization mode on a titanium (Ti) surface, was shown to enhance osteoblast growth and reduce fibroblast cell growth. This new method can securely attach a tailored interface to difficult materials such as Ti or ceramics. Here a more stable and higher density of NH₂ functional groups is able to withstand sterilization in ethanol. The biocompatibility, in terms of cell attachment and actin cytoskeleton development, was markedly improved in vitro, compared with untreated Ti surfaces and samples treated by other plasma modes. It gave a boosted (approximately six times higher) cellular response of osteoblasts in their initial adhesion stage. These factors should increase the formation of new bone around implants (reducing healing time), promoting osseointegration and thereby increasing implantation success rates. PMID:21370442

  6. Combining Phage and Yeast Cell Surface Antibody Display to Identify Novel Cell Type-Selective Internalizing Human Monoclonal Antibodies.

    Science.gov (United States)

    Bidlingmaier, Scott; Su, Yang; Liu, Bin

    2015-01-01

    Using phage antibody display, large libraries can be generated and screened to identify monoclonal antibodies with affinity for target antigens. However, while library size and diversity is an advantage of the phage display method, there is limited ability to quantitatively enrich for specific binding properties such as affinity. One way of overcoming this limitation is to combine the scale of phage display selections with the flexibility and quantitativeness of FACS-based yeast surface display selections. In this chapter we describe protocols for generating yeast surface antibody display libraries using phage antibody display selection outputs as starting material and FACS-based enrichment of target antigen-binding clones from these libraries. These methods should be widely applicable for the identification of monoclonal antibodies with specific binding properties. PMID:26060069

  7. Combined Flow Cytometric Measurement of Two Cell-Surface Antigens and DNA-RNA Content

    OpenAIRE

    sprotocols

    2014-01-01

    Author: Ingrid Schmid Corresponding author ([]()) ### INTRODUCTION Flow cytometry is frequently used to assess nucleic acid content in individual cells. Based on DNA content alone, however, cells in the quiescent G0 phase cannot be discriminated from cells in the proliferative G1 phase, as DNA content remains constant until S-phase entry. In contrast, by measuring RNA content in addition to DNA content, cells can be assigned to G0 and c...

  8. Cell surface engineering of Saccharomyces cerevisiae combined with membrane separation technology for xylitol production from rice straw hydrolysate.

    Science.gov (United States)

    Guirimand, Gregory; Sasaki, Kengo; Inokuma, Kentaro; Bamba, Takahiro; Hasunuma, Tomohisa; Kondo, Akihiko

    2016-04-01

    Xylitol, a value-added polyol deriving from D-xylose, is widely used in both the food and pharmaceutical industries. Despite extensive studies aiming to streamline the production of xylitol, the manufacturing cost of this product remains high while demand is constantly growing worldwide. Biotechnological production of xylitol from lignocellulosic waste may constitute an advantageous and sustainable option to address this issue. However, to date, there have been few reports of biomass conversion to xylitol. In the present study, xylitol was directly produced from rice straw hydrolysate using a recombinant Saccharomyces cerevisiae YPH499 strain expressing cytosolic xylose reductase (XR), along with β-glucosidase (BGL), xylosidase (XYL), and xylanase (XYN) enzymes (co-)displayed on the cell surface; xylitol production by this strain did not require addition of any commercial enzymes. All of these enzymes contributed to the consolidated bioprocessing (CBP) of the lignocellulosic hydrolysate to xylitol to produce 5.8 g/L xylitol with 79.5 % of theoretical yield from xylose contained in the biomass. Furthermore, nanofiltration of the rice straw hydrolysate provided removal of fermentation inhibitors while simultaneously increasing sugar concentrations, facilitating high concentration xylitol production (37.9 g/L) in the CBP. This study is the first report (to our knowledge) of the combination of cell surface engineering approach and membrane separation technology for xylitol production, which could be extended to further industrial applications. PMID:26631184

  9. Variations in Cell Surfaces of Estrogen Treated Breast Cancer Cells Detected by A Combined Instrument for Far-Field and Near-Field Microscopy

    Directory of Open Access Journals (Sweden)

    P. Perner

    2002-01-01

    Full Text Available The response of single breast cancer cells (cell line T‐47D to 17β‐estradiol (E2 under different concentrations was studied by using an instrument that allows to combine far‐field light microscopy with high resolution scanning near‐field (AFM/SNOM microscopy on the same cell. Different concentrations of E2 induce clearly different effects as well on cellular shape (in classical bright‐field imaging as on surface topography (atomic force imaging and absorbance (near‐field light transmission imaging. The differences range from a polygonal shape at zero via a roughly spherical shape at physiological up to a spindle‐like shape at un‐physiologically high concentrations. The surface topography of untreated control cells was found to be regular and smooth with small overall height modulations. At physiological E2 concentrations the surfaces became increasingly jagged as detected by an increase in membrane height. After application of the un‐physiological high E2 concentration the cell surface structures appeared to be smoother again with an irregular fine structure. The general behaviour of dose dependent differences was also found in the near‐field light transmission images. In order to quantify the treatment effects, line scans through the normalised topography images were drawn and a rate of co‐localisation between high topography and high transmission areas was calculated. The cell biological aspects of these observations are, so far, not studied in detail but measurements on single cells offer new perspectives to be empirically used in diagnosis and therapy control of breast cancers.

  10. Combination of photothermal and photodynamic inactivation of cancer cells through surface plasmon resonance of a gold nanoring

    Science.gov (United States)

    Chu, Chih-Ken; Tu, Yi-Chou; Hsiao, Jen-Hung; Yu, Jian-He; Yu, Chih-Kang; Chen, Shih-Yang; Tseng, Po-Hao; Chen, Shuai; Kiang, Yean-Woei; Yang, C. C.

    2016-03-01

    We demonstrate effective inactivation of oral cancer cells SAS through a combination of photothermal therapy (PTT) and photodynamic therapy (PDT) effects based on localized surface plasmon resonance (LSPR) around 1064 nm in wavelength of a Au nanoring (NRI) under femtosecond (fs) laser illumination. The PTT effect is caused by the LSPR-enhanced absorption of the Au NRI. The PDT effect is generated by linking the Au NRI with the photosensitizer of sulfonated aluminum phthalocyanines (AlPcS) for producing singlet oxygen through the LSPR-enhanced two-photon absorption (TPA) excitation of AlPcS. The laser threshold intensity for cancer cell inactivation with the applied Au NRI linked with AlPcS is significantly lower when compared to that with the Au NRI not linked with AlPcS. The comparison of inactivation threshold intensity between the cases of fs and continuous laser illuminations at the same wavelength and with the same average power confirms the crucial factor of TPA under fs laser illumination for producing the PDT effect.

  11. Mass Spectrometric Analysis of the Cell Surface N-Glycoproteome by Combining Metabolic Labeling and Click Chemistry

    Science.gov (United States)

    Smeekens, Johanna M.; Chen, Weixuan; Wu, Ronghu

    2015-04-01

    Cell surface N-glycoproteins play extraordinarily important roles in cell-cell communication, cell-matrix interactions, and cellular response to environmental cues. Global analysis is exceptionally challenging because many N-glycoproteins are present at low abundances and effective separation is difficult to achieve. Here, we have developed a novel strategy integrating metabolic labeling, copper-free click chemistry, and mass spectrometry (MS)-based proteomics methods to analyze cell surface N-glycoproteins comprehensively and site-specifically. A sugar analog containing an azido group, N-azidoacetylgalactosamine, was fed to cells to label glycoproteins. Glycoproteins with the functional group on the cell surface were then bound to dibenzocyclooctyne-sulfo-biotin via copper-free click chemistry under physiological conditions. After protein extraction and digestion, glycopeptides with the biotin tag were enriched by NeutrAvidin conjugated beads. Enriched glycopeptides were deglycosylated with peptide- N-glycosidase F in heavy-oxygen water, and in the process of glycan removal, asparagine was converted to aspartic acid and tagged with 18O for MS analysis. With this strategy, 144 unique N-glycopeptides containing 152 N-glycosylation sites were identified in 110 proteins in HEK293T cells. As expected, 95% of identified glycoproteins were membrane proteins, which were highly enriched. Many sites were located on important receptors, transporters, and cluster of differentiation proteins. The experimental results demonstrated that the current method is very effective for the comprehensive and site-specific identification of the cell surface N-glycoproteome and can be extensively applied to other cell surface protein studies.

  12. Combined Effects of Surface Morphology and Mechanical Straining Magnitudes on the Differentiation of Mesenchymal Stem Cells without Using Biochemical Reagents

    Directory of Open Access Journals (Sweden)

    Ji-Yeon Jang

    2011-01-01

    Full Text Available Existing studies examining the control of mesenchymal stem cell (MSC differentiation into desired cell types have used a variety of biochemical reagents such as growth factors despite possible side effects. Recently, the roles of biomimetic microphysical environments have drawn much attention in this field. We studied MSC differentiation and changes in gene expression in relation to osteoblast-like cell and smooth muscle-like cell type resulting from various microphysical environments, including differing magnitudes of tensile strain and substrate geometries for 8 days. In addition, we also investigated the residual effects of those selected microphysical environment factors on the differentiation by ceasing those factors for 3 days. The results of this study showed the effects of the strain magnitudes and surface geometries. However, the genes which are related to the same cell type showed different responses depending on the changes in strain magnitude and surface geometry. Also, different responses were observed three days after the straining was stopped. These data confirm that controlling microenvironments so that they mimic those in vivo contributes to the differentiation of MSCs into specific cell types. And duration of straining engagement was also found to play important roles along with surface geometry.

  13. Identification of microbes from the surfaces of food-processing lines based on the flow cytometric evaluation of cellular metabolic activity combined with cell sorting.

    Science.gov (United States)

    Juzwa, W; Duber, A; Myszka, K; Białas, W; Czaczyk, K

    2016-09-01

    In this study the design of a flow cytometry-based procedure to facilitate the detection of adherent bacteria from food-processing surfaces was evaluated. The measurement of the cellular redox potential (CRP) of microbial cells was combined with cell sorting for the identification of microorganisms. The procedure enhanced live/dead cell discrimination owing to the measurement of the cell physiology. The microbial contamination of the surface of a stainless steel conveyor used to process button mushrooms was evaluated in three independent experiments. The flow cytometry procedure provided a step towards monitoring of contamination and enabled the assessment of microbial food safety hazards by the discrimination of active, mid-active and non-active bacterial sub-populations based on determination of their cellular vitality and subsequently single cell sorting to isolate microbial strains from discriminated sub-populations. There was a significant correlation (r = 0.97; p < 0.05) between the bacterial cell count estimated by the pour plate method and flow cytometry, despite there being differences in the absolute number of cells detected. The combined approach of flow cytometric CRP measurement and cell sorting allowed an in situ analysis of microbial cell vitality and the identification of species from defined sub-populations, although the identified microbes were limited to culturable cells. PMID:27406324

  14. Identification of microbes from the surfaces of food-processing lines based on the flow cytometric evaluation of cellular metabolic activity combined with cell sorting.

    Science.gov (United States)

    Juzwa, W; Duber, A; Myszka, K; Białas, W; Czaczyk, K

    2016-09-01

    In this study the design of a flow cytometry-based procedure to facilitate the detection of adherent bacteria from food-processing surfaces was evaluated. The measurement of the cellular redox potential (CRP) of microbial cells was combined with cell sorting for the identification of microorganisms. The procedure enhanced live/dead cell discrimination owing to the measurement of the cell physiology. The microbial contamination of the surface of a stainless steel conveyor used to process button mushrooms was evaluated in three independent experiments. The flow cytometry procedure provided a step towards monitoring of contamination and enabled the assessment of microbial food safety hazards by the discrimination of active, mid-active and non-active bacterial sub-populations based on determination of their cellular vitality and subsequently single cell sorting to isolate microbial strains from discriminated sub-populations. There was a significant correlation (r = 0.97; p method and flow cytometry, despite there being differences in the absolute number of cells detected. The combined approach of flow cytometric CRP measurement and cell sorting allowed an in situ analysis of microbial cell vitality and the identification of species from defined sub-populations, although the identified microbes were limited to culturable cells.

  15. Combined intermittent hypoxia and surface muscle electrostimulation as a method to increase peripheral blood progenitor cell concentration

    Directory of Open Access Journals (Sweden)

    Azqueta Carmen

    2009-10-01

    Full Text Available Abstract Background Our goal was to determine whether short-term intermittent hypoxia exposure, at a level well tolerated by healthy humans and previously shown by our group to increase EPO and erythropoiesis, could mobilize hematopoietic stem cells (HSC and increase their presence in peripheral circulation. Methods Four healthy male subjects were subjected to three different protocols: one with only a hypoxic stimulus (OH, another with a hypoxic stimulus plus muscle electrostimulation (HME and the third with only muscle electrostimulation (OME. Intermittent hypobaric hypoxia exposure consisted of only three sessions of three hours at barometric pressure 540 hPa (equivalent to an altitude of 5000 m for three consecutive days, whereas muscular electrostimulation was performed in two separate periods of 25 min in each session. Blood samples were obtained from an antecubital vein on three consecutive days immediately before the experiment and 24 h, 48 h, 4 days and 7 days after the last day of hypoxic exposure. Results There was a clear increase in the number of circulating CD34+ cells after combined hypobaric hypoxia and muscular electrostimulation. This response was not observed after the isolated application of the same stimuli. Conclusion Our results open a new application field for hypobaric systems as a way to increase efficiency in peripheral HSC collection.

  16. Perylene Bisimide as a Promising Zinc Oxide Surface Modifier: Enhanced Interfacial Combination for Highly Efficient Inverted Polymer Solar Cells.

    Science.gov (United States)

    Nian, Li; Zhang, Wenqiang; Wu, Siping; Qin, Leiqiang; Liu, Linlin; Xie, Zengqi; Wu, Hongbin; Ma, Yuguang

    2015-11-25

    We report the application of a perylene bisimide (PBI-H) as zinc oxide (ZnO) surface modifier to afford an organic-inorganic co-interlayer for highly efficient inverted organic photovoltaics (i-OPV). By thermal annealing, a N-Zn chemical bond formed between PBI-H and ZnO, inducing close organic-inorganic combination. In addition, this co-interlayer shows decreased work function and increased electron transportation and conductivity, which are benefits for the cathode to enhance charge extraction efficiency and decrease recombination losses. As a result a highly efficient i-OPV was achieved with a power conversion efficiency (PCE) of 9.43% based on this co-interlayer with PTB7:PC71BM as the active layer, which shows over 35% enhancement compared to that of the device without the PBI-H layer. Moreover, this co-interlayer was widely applicable for i-OPVs based on various material systems, such as P3HT:PC61BM and PTB7-Th:PC71BM, resulting in PCE as high as 4.78% and 10.31%, respectively.

  17. Programming Surface Chemistry with Engineered Cells.

    Science.gov (United States)

    Zhang, Ruihua; Heyde, Keith C; Scott, Felicia Y; Paek, Sung-Ho; Ruder, Warren C

    2016-09-16

    We have developed synthetic gene networks that enable engineered cells to selectively program surface chemistry. E. coli were engineered to upregulate biotin synthase, and therefore biotin synthesis, upon biochemical induction. Additionally, two different functionalized surfaces were developed that utilized binding between biotin and streptavidin to regulate enzyme assembly on programmable surfaces. When combined, the interactions between engineered cells and surfaces demonstrated that synthetic biology can be used to engineer cells that selectively control and modify molecular assembly by exploiting surface chemistry. Our system is highly modular and has the potential to influence fields ranging from tissue engineering to drug development and delivery.

  18. Combination of pneumococcal surface protein A (PspA with whole cell pertussis vaccine increases protection against pneumococcal challenge in mice.

    Directory of Open Access Journals (Sweden)

    Maria Leonor S Oliveira

    Full Text Available Streptococcus pneumoniae is the leading cause of respiratory acute infections around the world. In Latin America, approximately 20,000 children under 5 years of age die of pneumococcal diseases annually. Pneumococcal surface protein A (PspA is among the best-characterized pneumococcal antigens that confer protection in animal models of pneumococcal infections and, as such, is a good alternative for the currently available conjugated vaccines. Efficient immune responses directed to PspA in animal models have already been described. Nevertheless, few low cost adjuvants for a subunit pneumococcal vaccine have been proposed to date. Here, we have tested the adjuvant properties of the whole cell Bordetella pertussis vaccine (wP that is currently part of the DTP (diphtheria-tetanus-pertussis vaccine administrated to children in several countries, as an adjuvant to PspA. Nasal immunization of BALB/c mice with a combination of PspA5 and wP or wP(low--a new generation vaccine that contains low levels of B. pertussis LPS--conferred protection against a respiratory lethal challenge with S. pneumoniae. Both PspA5-wP and PspA5-wP(low vaccines induced high levels of systemic and mucosal antibodies against PspA5, with similar profile, indicating no essential requirement for B. pertussis LPS in the adjuvant properties of wP. Accordingly, nasal immunization of C3H/HeJ mice with PspA5-wP conferred protection against the pneumococcal challenge, thus ruling out a role for TLR4 responses in the adjuvant activity and the protection mechanisms triggered by the vaccines. The high levels of anti-PspA5 antibodies correlated with increased cross-reactivity against PspAs from different clades and also reflected in cross-protection. In addition, passive immunization experiments indicated that antibodies played an important role in protection in this model. Finally, subcutaneous immunization with a combination of PspA5 with DTP(low protected mice against challenge with two

  19. Combined surface-enhanced Raman spectroscopy biotags and microfluidic platform for quantitative ratiometric discrimination between noncancerous and cancerous cells in flow

    Science.gov (United States)

    Pallaoro, Alessia; Hoonejani, Mehran R.; Braun, Gary B.; Meinhart, Carl; Moskovits, Martin

    2013-01-01

    Surface-enhanced Raman spectroscopy (SERS) biotags (SBTs) that carry peptides as cell recognition moieties were made from polymer-encapsulated silver nanoparticle dimers, infused with unique Raman reporter molecules. We previously demonstrated their potential use for identification of malignant cells, a central goal in cancer research, through a multiplexed, ratiometric method that can confidently distinguish between cancerous and noncancerous epithelial prostate cells in vitro based on receptor overexpression. Progress has been made toward the application of this quantitative methodology for the identification of cancer cells in a microfluidic flow-focusing device. Beads are used as cell mimics to evaluate the devices. Cells (and beads) are simultaneously incubated with two sets of SBTs while in suspension, then injected into the device for laser interrogation under flow. Each cell event is characterized by a composite Raman spectrum, deconvoluted into its single components to ultimately determine their relative contribution. We have found that using SBTs ratiometrically can provide cell identification in flow, insensitive to normal causes of uncertainty in optical measurements such as variations in focal plane, cell concentration, autofluorescence, and turbidity.

  20. Combined Effect of Surface Nano-Topography and Delivery of Therapeutics on the Adhesion of Tumor Cells on Porous Silicon Substrates

    KAUST Repository

    De Vitis, S.

    2016-02-23

    Porous silicon is a nano material in which pores with different sizes, densities and depths are infiltrated in conventional silicon imparting it augmented properties including biodegradability, biocompatibility, photoluminescence. Here, we realized porous silicon substrates in which the pore size and the fractal dimension were varied over a significant range. We loaded the described substrates with a PtCl(O, O′ − acac)(DMSO) antitumor drug and determined its release profile as a function of pore size over time up to 15 days. We observed that the efficacy of delivery augments with the pore size moving from small (∼ 8nm, efficiency of delivery ∼ 0.2) to large (∼ 55nm, efficiency of delivery ∼ 0.7). Then, we verified the adhesion of MCF-7 breast cancer cells on the described substrates with and without the administration of the antitumor drug. This permitted to decouple and understand the coincidental effects of nano-topography and a controlled dosage of drugs on cell adhesion and growth. While large pore sizes guarantee elevated drug dosages, large fractal dimensions boost cell adhesion on a surface. For the particular case of tumor cells and the delivery of an anti-tumor drug, substrates with a small fractal dimension and large pore size hamper cell growth. The competition between nano-topography and a controlled dosage of drugs may either accelerate or block the adhesion of cells on a nanostructured surface, for applications in tissue engineering, regenerative medicine, personalized lab-on-a-chips, and the rational design of implantable drug delivery systems.

  1. Wettability influences cell behavior on superhydrophobic surfaces with different topographies

    NARCIS (Netherlands)

    Lourenco, B.N.; Marchioli, G.; Song, W; Reis, R.L.; Blitterswijk, van C.A.; Karperien, H.B.J.; Apeldoorn, van A.A.; Mano, J.F.

    2012-01-01

    Surface wettability and topography are recognized as critical factors influencing cell behavior on biomaterials. So far only few works have reported cell responses on surfaces exhibiting extreme wettability in combination with surface topography. The goal of this work is to study whether cell behavi

  2. The Plant Cell Surface

    Institute of Scientific and Technical Information of China (English)

    Anne-Mie C.Emons; Kurt V.Fagerstedt

    2010-01-01

    @@ Multicellular organization and tissue construction has evolved along essentially different lines in plants and animals. Since plants do not run away, but are anchored in the soil, their tissues are more or less firm and stiff. This strength stems from the cell walls, which encase the fragile cytoplasm, and protect it.

  3. Combined surface and volumetric occlusion shading

    KAUST Repository

    Schott, Matthias O.

    2012-02-01

    In this paper, a method for interactive direct volume rendering is proposed that computes ambient occlusion effects for visualizations that combine both volumetric and geometric primitives, specifically tube shaped geometric objects representing streamlines, magnetic field lines or DTI fiber tracts. The proposed algorithm extends the recently proposed Directional Occlusion Shading model to allow the rendering of those geometric shapes in combination with a context providing 3D volume, considering mutual occlusion between structures represented by a volume or geometry. © 2012 IEEE.

  4. The cell-surface interaction.

    Science.gov (United States)

    Hayes, J S; Czekanska, E M; Richards, R G

    2012-01-01

    The realm of surface-dependent cell and tissue responses is the foundation of orthopaedic-device-related research. However, to design materials that elicit specific responses from tissues is a complex proposition mainly because the vast majority of the biological principles controlling the interaction of cells with implants remain largely ambiguous. Nevertheless, many surface properties, such as chemistry and topography, can be manipulated in an effort to selectively control the cell-material interaction. On the basis of this information there has been much research in this area, including studies focusing on the structure and composition of the implant interface, optimization of biological and chemical coatings and elucidation of the mechanisms involved in the subsequent cell-material interactions. Although a wealth of information has emerged, it also advocates the complexity and dynamism of the cell-material interaction. Therefore, this chapter aims to provide the reader with an introduction to the basic concepts of the cell-material interaction and to provide an insight into the factors involved in determining the cell and tissue response to specific surface features, with specific emphasis on surface microtopography. PMID:21984613

  5. Improved cell mediated immune responses after successful re-vaccination of non-responders to the hepatitis B virus surface antigen (HBsAg) vaccine using the combined hepatitis A and B vaccine.

    Science.gov (United States)

    Nyström, Jessica; Cardell, Kristina; Björnsdottir, Thora Björg; Fryden, Aril; Hultgren, Catharina; Sällberg, Matti

    2008-11-01

    We successfully re-vaccinated hepatitis B virus (HBV) vaccine non-responders using a double dose of the combined hepatitis A virus (HAV) and HBV vaccine. The hope was to improve priming of hepatitis B surface antigen (HBsAg)-specific cell mediated immune response (CMI) by an increased antigen dose and a theoretical adjuvant-effect from the local presence of a HAV-specific CMI. A few non-responders had a detectable HBsAg-specific CMI before re-vaccination. An in vitro detectable HBsAg-specific CMI was primed equally effective in non-responders (58%) as in first time vaccine recipients (68%). After the third dose a weak, albeit significant, association was observed between the magnitude of HBsAg-specific proliferation and anti-HBs levels. This regimen improves the priming of HBsAg-specific CMIs and antibodies.

  6. Magnetic resonance force microscopy combined with surface topography

    Science.gov (United States)

    Tsuji, S.; Yoshinari, Y.; Kawai, E.; Nakajima, K.; Park, H. S.; Shindo, D.

    2007-10-01

    A new method of surface microscopy is proposed, which combines three-dimensional electron spin resonance imaging by magnetic resonance force microscopy (MRFM) and topographic imaging of the sample surface by scanning force microscopy (SFM). In order to demonstrate its potential for the identification of microscale objects, the individual and combined images are used to provide the locations, shapes and spin density distributions of target phantom objects. We report spatial resolution in MRFM of 2.8 × 2.8 × 2.0μm 3. This could be improved to the theoretical limit of 0.08 × 0.08 × 0.04μm 3 through reduction of the thermal noise by cooling to cryogenic temperatures ˜0.5 K. We believe that this type of microscopy will become a very useful tool for the investigation of anomalies induced in surfaces by materials buried below the surface.

  7. Magnetic Resonance Force Microscopy Combined with Surface Topography

    Science.gov (United States)

    Tsuji, Shigenori; Yoshinari, Yohsuke

    2007-03-01

    In this presentation, we will show magnetic resonance force microscopy imaging combined with surface topography. The individual and combined images taken in the same coordinate are presented for extraction of the position, shapes and spin density distribution of target phantoms. This imaging technique is useful applied when the surface needs to be investigated in relation to the influence of a material buried below the surface. In our method, the surface topography was observed by the AFM with tapping mode. The spin density distribution was measured by the MRFM with the cyclic saturation technique. The AFM and MRFM experiments were made one after another by using the same experimental set-up, and their images were merged together afterwards. The sample consists of two kind of materials, one is DPPH containing unpaird spins and the other is a glass bead. DPPH particles with the size of 5˜8 micrometer and a 8.8 micrometer single bead were glued on a commercial cantilever.

  8. The Cell Surface Proteome of Human Mesenchymal Stromal Cells

    OpenAIRE

    Christian Niehage; Charlotte Steenblock; Theresia Pursche; Martin Bornhäuser; Denis Corbeil; Bernard Hoflack

    2011-01-01

    BACKGROUND: Multipotent human mesenchymal stromal cells (hMSCs) are considered as promising biological tools for regenerative medicine. Their antibody-based isolation relies on the identification of reliable cell surface markers. METHODOLOGY/PRINCIPAL FINDINGS: To obtain a comprehensive view of the cell surface proteome of bone marrow-derived hMSCs, we have developed an analytical pipeline relying on cell surface biotinylation of intact cells using cell impermeable, cleavable sulfo-NHS-SS-bio...

  9. A mass spectrometric-derived cell surface protein atlas.

    Directory of Open Access Journals (Sweden)

    Damaris Bausch-Fluck

    Full Text Available Cell surface proteins are major targets of biomedical research due to their utility as cellular markers and their extracellular accessibility for pharmacological intervention. However, information about the cell surface protein repertoire (the surfaceome of individual cells is only sparsely available. Here, we applied the Cell Surface Capture (CSC technology to 41 human and 31 mouse cell types to generate a mass-spectrometry derived Cell Surface Protein Atlas (CSPA providing cellular surfaceome snapshots at high resolution. The CSPA is presented in form of an easy-to-navigate interactive database, a downloadable data matrix and with tools for targeted surfaceome rediscovery (http://wlab.ethz.ch/cspa. The cellular surfaceome snapshots of different cell types, including cancer cells, resulted in a combined dataset of 1492 human and 1296 mouse cell surface glycoproteins, providing experimental evidence for their cell surface expression on different cell types, including 136 G-protein coupled receptors and 75 membrane receptor tyrosine-protein kinases. Integrated analysis of the CSPA reveals that the concerted biological function of individual cell types is mainly guided by quantitative rather than qualitative surfaceome differences. The CSPA will be useful for the evaluation of drug targets, for the improved classification of cell types and for a better understanding of the surfaceome and its concerted biological functions in complex signaling microenvironments.

  10. A mass spectrometric-derived cell surface protein atlas.

    Science.gov (United States)

    Bausch-Fluck, Damaris; Hofmann, Andreas; Bock, Thomas; Frei, Andreas P; Cerciello, Ferdinando; Jacobs, Andrea; Moest, Hansjoerg; Omasits, Ulrich; Gundry, Rebekah L; Yoon, Charles; Schiess, Ralph; Schmidt, Alexander; Mirkowska, Paulina; Härtlová, Anetta; Van Eyk, Jennifer E; Bourquin, Jean-Pierre; Aebersold, Ruedi; Boheler, Kenneth R; Zandstra, Peter; Wollscheid, Bernd

    2015-01-01

    Cell surface proteins are major targets of biomedical research due to their utility as cellular markers and their extracellular accessibility for pharmacological intervention. However, information about the cell surface protein repertoire (the surfaceome) of individual cells is only sparsely available. Here, we applied the Cell Surface Capture (CSC) technology to 41 human and 31 mouse cell types to generate a mass-spectrometry derived Cell Surface Protein Atlas (CSPA) providing cellular surfaceome snapshots at high resolution. The CSPA is presented in form of an easy-to-navigate interactive database, a downloadable data matrix and with tools for targeted surfaceome rediscovery (http://wlab.ethz.ch/cspa). The cellular surfaceome snapshots of different cell types, including cancer cells, resulted in a combined dataset of 1492 human and 1296 mouse cell surface glycoproteins, providing experimental evidence for their cell surface expression on different cell types, including 136 G-protein coupled receptors and 75 membrane receptor tyrosine-protein kinases. Integrated analysis of the CSPA reveals that the concerted biological function of individual cell types is mainly guided by quantitative rather than qualitative surfaceome differences. The CSPA will be useful for the evaluation of drug targets, for the improved classification of cell types and for a better understanding of the surfaceome and its concerted biological functions in complex signaling microenvironments. PMID:25894527

  11. Cells behaviors and genotoxicity on topological surface

    Energy Technology Data Exchange (ETDEWEB)

    Yang, N.; Yang, M.K.; Bi, S.X. [Tianjin Key Laboratory of Fiber Modification and Functional Fiber, School of Materials Science and Engineering, Tianjin Polytechnic University, Tianjin, 300387 (China); Chen, L., E-mail: chenlis@tjpu.edu.cn [Tianjin Key Laboratory of Fiber Modification and Functional Fiber, School of Materials Science and Engineering, Tianjin Polytechnic University, Tianjin, 300387 (China); Zhu, Z.Y.; Gao, Y.T.; Du, Z. [Tianjin Key Laboratory of Artificial Cell, Tianjin Third Central Hospital, Tianjin, 300170 (China)

    2013-08-01

    To investigate different cells behaviors and genotoxicity, which were driven by specific microenvironments, three patterned surfaces (pillars, wide grooves and narrow grooves) and one smooth surface were prepared by template-based technique. Vinculin is a membrane-cytoskeletal protein in focal adhesion plaques and associates with cell–cell and cell–matrix junctions, which can promote cell adhesion and spreading. The immunofluorescence staining of vinculin revealed that the narrow grooves patterned substrate was favorable for L929 cell adhesion. For cell multiplication, the narrow grooves surface was fitted for the proliferation of L929, L02 and MSC cells, the pillars surface was only in favor of L929 cells to proliferate during 7 days of cell cultivation. Cell genetic toxicity was evaluated by cellular micronuclei test (MNT). The results indicated that topological surfaces were more suitable for L929 cells to proliferate and maintain the stability of genome. On the contrary, the narrow grooves surface induced higher micronuclei ratio of L02 and MSC cells than other surfaces. With the comprehensive results of cell multiplication and MNT, it was concluded that the wide grooves surface was best fitted for L02 cells to proliferate and have less DNA damages, and the smooth surface was optimum for the research of MSC cells in vitro. - Highlights: • Different cells behaviors on microstructure surfaces were discussed in this paper. • The expression of cell protein of Vinculin was studied in this research. • Cellular micronuclei test was applied to evaluate cells' genotoxicity. • Cell genotoxicity was first studied in the research field of topological surfaces.

  12. Combining multi-atlas segmentation with brain surface estimation

    Science.gov (United States)

    Huo, Yuankai; Carass, Aaron; Resnick, Susan M.; Pham, Dzung L.; Prince, Jerry L.; Landman, Bennett A.

    2016-03-01

    Whole brain segmentation (with comprehensive cortical and subcortical labels) and cortical surface reconstruction are two essential techniques for investigating the human brain. The two tasks are typically conducted independently, however, which leads to spatial inconsistencies and hinders further integrated cortical analyses. To obtain self-consistent whole brain segmentations and surfaces, FreeSurfer segregates the subcortical and cortical segmentations before and after the cortical surface reconstruction. However, this "segmentation to surface to parcellation" strategy has shown limitation in various situations. In this work, we propose a novel "multi-atlas segmentation to surface" method called Multi-atlas CRUISE (MaCRUISE), which achieves self-consistent whole brain segmentations and cortical surfaces by combining multi-atlas segmentation with the cortical reconstruction method CRUISE. To our knowledge, this is the first work that achieves the reliability of state-of-the-art multi-atlas segmentation and labeling methods together with accurate and consistent cortical surface reconstruction. Compared with previous methods, MaCRUISE has three features: (1) MaCRUISE obtains 132 cortical/subcortical labels simultaneously from a single multi-atlas segmentation before reconstructing volume consistent surfaces; (2) Fuzzy tissue memberships are combined with multi-atlas segmentations to address partial volume effects; (3) MaCRUISE reconstructs topologically consistent cortical surfaces by using the sulci locations from multi-atlas segmentation. Two data sets, one consisting of five subjects with expertly traced landmarks and the other consisting of 100 volumes from elderly subjects are used for validation. Compared with CRUISE, MaCRUISE achieves self-consistent whole brain segmentation and cortical reconstruction without compromising on surface accuracy. MaCRUISE is comparably accurate to FreeSurfer while achieving greater robustness across an elderly population.

  13. Functions of proteoglycans at the cell surface

    DEFF Research Database (Denmark)

    Höök, M; Woods, A; Johansson, S;

    1986-01-01

    Proteoglycans (primarily heparan sulphate proteoglycans) are found at the surface of most adherent eukaryotic cells. Earlier studies suggest that these molecules can be associated with the cell surface principally by two different mechanisms. Proteoglycans may occur as membrane-intercalated glyco......Proteoglycans (primarily heparan sulphate proteoglycans) are found at the surface of most adherent eukaryotic cells. Earlier studies suggest that these molecules can be associated with the cell surface principally by two different mechanisms. Proteoglycans may occur as membrane......-intercalated glycoproteins, where the core protein of the proteoglycan is anchored in the lipid interior of the plasma membrane, or they may be bound via the polysaccharide components of the molecule to specific anchoring proteins present at the cell surface. A number of functions have been proposed for cell surface...

  14. Probe microscopy: Scanning below the cell surface

    Science.gov (United States)

    Sahin, Ozgur

    2008-08-01

    Conventional atomic force microscopy probes only the surface of specimens. A related technique called scanning near-field ultrasonic holography can now image nanoparticles buried below the surfaces of cells, which could prove useful in nanotoxicology.

  15. The pancreatic beta cell surface proteome

    OpenAIRE

    Stützer, I.; Esterházy, D.; Stoffel, M.

    2012-01-01

    The pancreatic beta cell is responsible for maintaining normoglycaemia by secreting an appropriate amount of insulin according to blood glucose levels. The accurate sensing of the beta cell extracellular environment is therefore crucial to this endocrine function and is transmitted via its cell surface proteome. Various surface proteins that mediate or affect beta cell endocrine function have been identified, including growth factor and cytokine receptors, transporters, ion channels and prote...

  16. Multi-scale cell/surface interaction on modified titanium aluminum vanadium surfaces

    Science.gov (United States)

    Chen, Jianbo

    This dissertation presents a series of experimental studies of the effects of multi-scale cell/surface interactions on modified Ti-6Al-4V surfaces. These include laser-grooved surfaces; porous structures and RGD-coated laser-grooved surfaces. A nano-second DPSS UV lasers with a Gaussian pulse energy profile was used to introduce the desired micro-groove geometries onto Ti-6Al-4V surfaces. This was done without inducing micro-cracks or significant changes in surface chemistry within the heat affected zones. The desired 8-12 mum groove depths and widths were achieved by the control of pulse frequency, scan speed, and the lens focal length that controls spot size. The interactions between human osteosarcoma (HOS) cells and laser-grooved Ti-6Al-4V surfaces were investigated after 48 hours of cell culture. The cell behavior, including cell spreading, alignment and adhesion, was elucidated using scanning electronic microscopy (SEM), immuno-fluorescence staining and enzymatic detachment. Contact guidance was shown to increase as grooved spacing decreased. For the range of micro-groove geometries studied, micro-grooves with groove spacings of 20 mum provided the best combination of cell orientation and adhesion. Short-term adhesion experiments (15 mins to 1 day) also revealed that there is a positive correlation between cell orientation and cell adhesion. Contact guidance on the micro-grooved surfaces is shown to be enhanced by nano- and micro-scale asperities that provide sites for the attachment of lamellopodia during cell locomotion and spreading. Contact guidance is also promoted by the geometrical confinement provided by laser grooves. An experimental study of initial cell spreading and ingrowth into Ti-6Al-4V porous structures was also carried out on porous structures with different pore sizes and geometries. A combination of SEM, the tetrazolium salt (MTT) colorimetric assay and enzymatic detachment were used to study cell spreading and adhesion. The extent of cell

  17. Surface Functionalization for Protein and Cell Patterning

    Science.gov (United States)

    Colpo, Pascal; Ruiz, Ana; Ceriotti, Laura; Rossi, François

    The interaction of biological systems with synthetic material surfaces is an important issue for many biological applications such as implanted devices, tissue engineering, cell-based sensors and assays, and more generally biologic studies performed ex vivo. To ensure reliable outcomes, the main challenge resides in the ability to design and develop surfaces or artificial micro-environment that mimic 'natural environment' in interacting with biomolecules and cells without altering their function and phenotype. At this effect, microfabrication, surface chemistry and material science play a pivotal role in the design of advanced in-vitro systems for cell culture applications. In this chapter, we discuss and describe different techniques enabling the control of cell-surface interactions, including the description of some techniques for immobilization of ligands for controlling cell-surface interactions and some methodologies for the creation of well confined cell rich areas.

  18. Surface Plasmon Resonance for Cell-Based Clinical Diagnosis

    Directory of Open Access Journals (Sweden)

    Yuhki Yanase

    2014-03-01

    Full Text Available Non-invasive real-time observations and the evaluation of living cell conditions and functions are increasingly demanded in life sciences. Surface plasmon resonance (SPR sensors detect the refractive index (RI changes on the surface of sensor chips in label-free and on a real-time basis. Using SPR sensors, we and other groups have developed techniques to evaluate living cells’ reactions in response to stimuli without any labeling in a real-time manner. The SPR imaging (SPRI system for living cells may visualize single cell reactions and has the potential to expand application of SPR cell sensing for clinical diagnosis, such as multi-array cell diagnostic systems and detection of malignant cells among normal cells in combination with rapid cell isolation techniques.

  19. Controlled surface chemistries and quantitative cell response

    Science.gov (United States)

    Plant, Anne L.

    2002-03-01

    Living cells experience a large number of signaling cues from their extracellular matrix. As a result of these inputs, a variety of intracellular signaling pathways are apparently initiated simultaneously. The vast array of alternative responses that result from the integration of these inputs suggests that it may be reasonable to look for cellular response not as an 'on' or 'off' condition but as a distribution of responses. A difficult challenge is to determine whether variations in responses from individual cells arise from the complexity of intracellular signals or are due to variations in the cell culture environment. By controlling surface chemistry so that every cell 'sees' the same chemical and physical environment, we can begin to assess how the distribution of cell response is affected strictly by changes in the chemistry of the cell culture surface. Using the gene for green fluorescent protein linked to the gene for the promoter of the extracellular matrix protein, tenascin, we can easily probe the end product in a signaling pathway that is purported to be linked to surface protein chemistry and to cell shape. Cell response to well-controlled, well-characterized, and highly reproducible surfaces prepared using soft lithography techniques are compared with more conventional ways of preparing extracellular matrix proteins for cell culture. Using fluorescence microscopy and image analysis of populations of cells on these surfaces, we probe quantitatively the relationship between surface chemistry, cell shape and variations in gene expression endpoint.

  20. The cell surface of Trypanosoma cruzi

    Directory of Open Access Journals (Sweden)

    Wanderley de Souza

    1984-01-01

    Full Text Available The cell surface of trypanosomatids is formed by the plasma membrane and a layer of sub-pellicular microtubules which are connected to the plasma membrane. The plasma membrane is composed by proteins, lipids and carbohydrates which form the glycocalix. In this paper we will review briefly aspects related to the organization of the cell surface of Trypanosoma cruzi.

  1. Radioimmunoassay to quantitatively measure cell surface immunoglobulins

    International Nuclear Information System (INIS)

    A radioimmunoassay techniques developed to quantitatively measure the presence of immunoglobulins on the surface of cells, is described. The amount of immunoglobulins found on different tumor cells varied from 200 to 1140 ng/106 cells. Determination of immunoglobulins on the peripheral lymphocytes obtained from different cancer patients varied between 340 to 1040 ng/106 cells. Cultured tumor cells, on the other hand, were found to contain negligible quantities of human IgG

  2. Cell-surface remodelling during mammalian erythropoiesis.

    Science.gov (United States)

    Wraith, D C; Chesterton, C J

    1982-10-15

    Current evidence suggests that the major cell-surface modification occurring during mammalian erythropoiesis could be generated by two separate mechanisms: either selective loss of membrane proteins during enucleation or endocytosis at the subsequent reticulocyte and erythrocyte stages. The former idea was tested by collecting developing rabbit erythroid cells before and after the enucleation step and comparing their cell-surface protein composition via radiolabelling and electrophoresis. Few changes were observed. Our data thus lend support to the endocytosis mechanism.

  3. Nanostructuring of Solar Cell Surfaces

    DEFF Research Database (Denmark)

    Davidsen, Rasmus Schmidt; Schmidt, Michael Stenbæk

    schemes such as atomic layer deposition (ALD) of Al2O3. ALD Al2O3 passivation on black Si yields surface recombination velocity (SRV) below 80 cm/s and implied open-circuit voltage (iVOC) of 680 mV. Surface recombination velocity of 20 cm/s and implied open-circuit voltage of 695 mV is obtained for black...

  4. Implant Surface Design Regulates Mesenchymal Stem Cell Differentiation and Maturation.

    Science.gov (United States)

    Boyan, B D; Cheng, A; Olivares-Navarrete, R; Schwartz, Z

    2016-03-01

    Changes in dental implant materials, structural design, and surface properties can all affect biological response. While bulk properties are important for mechanical stability of the implant, surface design ultimately contributes to osseointegration. This article reviews the surface parameters of dental implant materials that contribute to improved cell response and osseointegration. In particular, we focus on how surface design affects mesenchymal cell response and differentiation into the osteoblast lineage. Surface roughness has been largely studied at the microscale, but recent studies have highlighted the importance of hierarchical micron/submicron/nanosurface roughness, as well as surface roughness in combination with surface wettability. Integrins are transmembrane receptors that recognize changes in the surface and mediate downstream signaling pathways. Specifically, the noncanonical Wnt5a pathway has been implicated in osteoblastic differentiation of cells on titanium implant surfaces. However, much remains to be elucidated. Only recently have studies been conducted on the differences in biological response to implants based on sex, age, and clinical factors; these all point toward differences that advocate for patient-specific implant design. Finally, challenges in implant surface characterization must be addressed to optimize and compare data across studies. An understanding of both the science and the biology of the materials is crucial for developing novel dental implant materials and surface modifications for improved osseointegration.

  5. Cell behaviour on chemically microstructured surfaces

    Energy Technology Data Exchange (ETDEWEB)

    Magnani, Agnese; Priamo, Alfredo; Pasqui, Daniela; Barbucci, Rolando

    2003-03-03

    Micropatterned surfaces with different chemical topographies were synthesised in order to investigate the influence of surface chemistry and topography on cell behaviour. The microstructured materials were synthesised by photoimmobilising natural Hyaluronan (Hyal) and its sulphated derivative (HyalS), both adequately functionalised with a photorective moiety, on glass substrates. Four different grating patterns (10, 25, 50 and 100 {mu}m) were used to pattern the hyaluronan. The micropatterned samples were analysed by Secondary Ions Mass Spectrometry, Scanning Electron Microscopy (SEM) and Atomic Force Microscopy to investigate the chemistry and the topography of the surfaces. The spectroscopic and microscopic analysis of the microstructured surfaces revealed that the photoimmobilisation process was successful, demonstrating that the photomask patterns were well reproduced on the sample surface. The influence of chemical topographies on the cell behaviour was then analysed. Human and 3T3 fibroblasts, bovine aortic and human (HGTFN line) endothelial cells were used and their behaviour on the micropatterned surfaces was analysed in terms of adhesion, proliferation, locomotion and orientation. Both chemical and topographical controls were found to be important for cell guidance. By decreasing the stripe dimensions, a more fusiform shape of cell was observed. At the same time, the cell locomotion and orientation parallel to the structure increased. However, differences in cell behaviour were detected according to both cell type and micropattern dimensions.

  6. The combined action of UV irradiation and chemical treatment on the titanium surface of dental implants

    International Nuclear Information System (INIS)

    Highlights: • A combined UV irradiation and H2O2 treatment was applied to titanium surfaces. • A thin, homogeneous, not porous, crack-free and bioactive oxide layer was obtained. • The process significantly improves the biological response of titanium surfaces. • A clinical case demonstrates the effectiveness of the proposed treatment. - Abstract: The purpose of this paper is to describe an innovative treatment for titanium dental implants, aimed at faster and more effective osteointegration. The treatment has been performed with the use of hydrogen peroxide, whose action was enhanced by concomitant exposure to a source of ultraviolet light. The developed surface oxide layer was characterized from the physical and chemical points of view. Moreover osteoblast-like SaOS2 cells were cultured on treated and control titanium surfaces and cell behavior investigated by scanning electron microscope observation and gene expression measurements. The described process produces, in only 6 min, a thin, homogeneous, not porous, free of cracks and bioactive (in vitro apatite precipitation) oxide layer. High cell density, peculiar morphology and overexpression of several genes involved with osteogenesis have been observed on modified surfaces. The proposed process significantly improves the biological response of titanium surfaces, and is an interesting solution for the improvement of bone integration of dental implants. A clinical application of the described surfaces, with a 5 years follow-up, is reported in the paper, as an example of the effectiveness of the proposed treatment

  7. The combined action of UV irradiation and chemical treatment on the titanium surface of dental implants

    Energy Technology Data Exchange (ETDEWEB)

    Spriano, Silvia [Politecnico di Torino, Department of Applied Science and Technology, Corso Duca degli Abruzzi, 24-10129 Torino (Italy); Ferraris, Sara, E-mail: sara.ferraris@polito.it [Politecnico di Torino, Department of Applied Science and Technology, Corso Duca degli Abruzzi, 24-10129 Torino (Italy); Bollati, Daniele; Morra, Marco; Cassinelli, Clara [Nobil Bio Ricerche, Portacomaro (Italy); Lorenzon, Giorgio [Centro Chirurgico, Via Mallonetto, 47, 10032, Brandizzo Torino (Italy)

    2015-09-15

    Highlights: • A combined UV irradiation and H{sub 2}O{sub 2} treatment was applied to titanium surfaces. • A thin, homogeneous, not porous, crack-free and bioactive oxide layer was obtained. • The process significantly improves the biological response of titanium surfaces. • A clinical case demonstrates the effectiveness of the proposed treatment. - Abstract: The purpose of this paper is to describe an innovative treatment for titanium dental implants, aimed at faster and more effective osteointegration. The treatment has been performed with the use of hydrogen peroxide, whose action was enhanced by concomitant exposure to a source of ultraviolet light. The developed surface oxide layer was characterized from the physical and chemical points of view. Moreover osteoblast-like SaOS2 cells were cultured on treated and control titanium surfaces and cell behavior investigated by scanning electron microscope observation and gene expression measurements. The described process produces, in only 6 min, a thin, homogeneous, not porous, free of cracks and bioactive (in vitro apatite precipitation) oxide layer. High cell density, peculiar morphology and overexpression of several genes involved with osteogenesis have been observed on modified surfaces. The proposed process significantly improves the biological response of titanium surfaces, and is an interesting solution for the improvement of bone integration of dental implants. A clinical application of the described surfaces, with a 5 years follow-up, is reported in the paper, as an example of the effectiveness of the proposed treatment.

  8. Cell orientation on a stripe-micropatterned surface

    Institute of Scientific and Technical Information of China (English)

    SUN JianGuo; TANG Jian; DING JianDong

    2009-01-01

    Stripe-micropatterned surfaces have recently been a unique tool to study cell orientation. In this paper,we prepared,by the photolithography transfer technique,stable gold (Au) micropatterns on PEG hydrogel surfaces with defined cell-resistant (PEG hydrogel) and cell-adhesive (gold microstripes) proparties. 3T3 fibroblasts were cultured on Au-microstripe surfaces to observe cell adhesion and orientation. Five statistical parameters were defined and used to describe cell orientation on micropatterns.With the increase of inter-stripe distance,the orientational order parameter,the ratio of long and short axes of a cell,and the occupation fraction of cells on stripes increased gradually,whereas the spreading area of a single cell decreased. The abrupt changes of these four parameters did not happen at the same inter-distance. The adhesion ratio of a cell on Au stripes over cell spreading area did not change monotonically as a function of inter-stripe distance. The combination of the 5 statistical parameters represented well the cell orientation behaviors semi-quantitatively.

  9. Functional dynamics of cell surface membrane proteins

    Science.gov (United States)

    Nishida, Noritaka; Osawa, Masanori; Takeuchi, Koh; Imai, Shunsuke; Stampoulis, Pavlos; Kofuku, Yutaka; Ueda, Takumi; Shimada, Ichio

    2014-04-01

    Cell surface receptors are integral membrane proteins that receive external stimuli, and transmit signals across plasma membranes. In the conventional view of receptor activation, ligand binding to the extracellular side of the receptor induces conformational changes, which convert the structure of the receptor into an active conformation. However, recent NMR studies of cell surface membrane proteins have revealed that their structures are more dynamic than previously envisioned, and they fluctuate between multiple conformations in an equilibrium on various timescales. In addition, NMR analyses, along with biochemical and cell biological experiments indicated that such dynamical properties are critical for the proper functions of the receptors. In this review, we will describe several NMR studies that revealed direct linkage between the structural dynamics and the functions of the cell surface membrane proteins, such as G-protein coupled receptors (GPCRs), ion channels, membrane transporters, and cell adhesion molecules.

  10. Responses of fibroblasts and glial cells to nanostructured platinum surfaces

    Science.gov (United States)

    Pennisi, C. P.; Sevcencu, C.; Dolatshahi-Pirouz, A.; Foss, M.; Lundsgaard Hansen, J.; Nylandsted Larsen, A.; Zachar, V.; Besenbacher, F.; Yoshida, K.

    2009-09-01

    The chronic performance of implantable neural prostheses is affected by the growth of encapsulation tissue onto the stimulation electrodes. Encapsulation is associated with activation of connective tissue cells at the electrode's metallic contacts, usually made of platinum. Since surface nanotopography can modulate the cellular responses to materials, the aim of the present work was to evaluate the 'in vitro' responses of connective tissue cells to platinum strictly by modulating its surface nanoroughness. Using molecular beam epitaxy combined with sputtering, we produced platinum nanostructured substrates consisting of irregularly distributed nanopyramids and investigated their effect on the proliferation, cytoskeletal organization and cellular morphology of primary fibroblasts and transformed glial cells. Cells were cultured on these substrates and their responses to surface roughness were studied. After one day in culture, the fibroblasts were more elongated and their cytoskeleton less mature when cultured on rough substrates. This effect increased as the roughness of the surface increased and was associated with reduced cell proliferation throughout the observation period (4 days). Morphological changes also occurred in glial cells, but they were triggered by a different roughness scale and did not affect cellular proliferation. In conclusion, surface nanotopography modulates the responses of fibroblasts and glial cells to platinum, which may be an important factor in optimizing the tissue response to implanted neural electrodes.

  11. Adhesion of cells to polystyrene surfaces

    OpenAIRE

    1983-01-01

    The surface treatment of polystyrene, which is required to make polystyrene suitable for cell adhesion and spreading, was investigated. Examination of surfaces treated with sulfuric acid or various oxidizing agents using (a) x-ray photoelectron and attenuated total reflection spectroscopy and (b) measurement of surface carboxyl-, hydroxyl-, and sulfur-containing groups by various radiochemical methods showed that sulfuric acid produces an insignificant number of sulfonic acid groups on polyst...

  12. Nanotomography of Cell Surfaces with Evanescent Fields

    Directory of Open Access Journals (Sweden)

    Michael Wagner

    2008-01-01

    Full Text Available The technique of variable-angle total internal reflection fluorescence microscopy (TIRFM and its application to nanotomography of cell surfaces are described. Present applications include (1 3D imaging of chromosomes in their metaphase to demonstrate axial resolution in the nanometre range, (2 measurements of cell-substrate topology, which upon cholesterol depletion shows some loosening of cell-substrate contacts, and (3 measurements of cell topology upon photodynamic therapy (PDT, which demonstrate cell swelling and maintenance of focal contacts. The potential of the method for in vitro diagnostics, but also some requirements and limitations are discussed.

  13. Combining surface reanalysis and remote sensing data for monitoring evapotranspiration

    Directory of Open Access Journals (Sweden)

    M. Marshall

    2012-02-01

    Full Text Available Climate change is expected to have the greatest impact on the world's poor. In the Sahel, a climatically sensitive region where rain-fed agriculture is the primary livelihood, expected decreases in water supply will increase food insecurity. Studies on climate change and the intensification of the water cycle in sub-Saharan Africa are few. This is due in part to poor calibration of modeled actual evapotranspiration (AET, a key input in continental-scale hydrologic models. In this study, a model driven by dynamic canopy AET was combined with the Global Land Data Assimilation System realization of the NOAH Land Surface Model (GNOAH wet canopy and soil AET for monitoring purposes in sub-Saharan Africa. The performance of the hybrid model was compared against AET from the GNOAH model and dynamic model using eight eddy flux towers representing major biomes of sub-Saharan Africa. The greatest improvements in model performance are at humid sites with dense vegetation, while performance at semi-arid sites is poor, but better than individual models. The reduction in errors using the hybrid model can be attributed to the integration of a dynamic vegetation component with land surface model estimates, improved model parameterization, and reduction of multiplicative effects of uncertain data.

  14. Combining surface reanalysis and remote sensing data for monitoring evapotranspiration

    Science.gov (United States)

    Marshall, M.; Tu, K.; Funk, C.; Michaelsen, J.; Williams, P.; Williams, C.; Ardö, J.; Marie, B.; Cappelaere, B.; Grandcourt, A.; Nickless, A.; Nouvellon, Y.; Scholes, R.; Kutsch, W.

    2012-02-01

    Climate change is expected to have the greatest impact on the world's poor. In the Sahel, a climatically sensitive region where rain-fed agriculture is the primary livelihood, expected decreases in water supply will increase food insecurity. Studies on climate change and the intensification of the water cycle in sub-Saharan Africa are few. This is due in part to poor calibration of modeled actual evapotranspiration (AET), a key input in continental-scale hydrologic models. In this study, a model driven by dynamic canopy AET was combined with the Global Land Data Assimilation System realization of the NOAH Land Surface Model (GNOAH) wet canopy and soil AET for monitoring purposes in sub-Saharan Africa. The performance of the hybrid model was compared against AET from the GNOAH model and dynamic model using eight eddy flux towers representing major biomes of sub-Saharan Africa. The greatest improvements in model performance are at humid sites with dense vegetation, while performance at semi-arid sites is poor, but better than individual models. The reduction in errors using the hybrid model can be attributed to the integration of a dynamic vegetation component with land surface model estimates, improved model parameterization, and reduction of multiplicative effects of uncertain data.

  15. Combinational Effect of Cell Adhesion Biomolecules and Their Immobilized Polymer Property to Enhance Cell-Selective Adhesion

    Directory of Open Access Journals (Sweden)

    Rio Kurimoto

    2016-01-01

    Full Text Available Although surface immobilization of medical devices with bioactive molecules is one of the most widely used strategies to improve biocompatibility, the physicochemical properties of the biomaterials significantly impact the activity of the immobilized molecules. Herein we investigate the combinational effects of cell-selective biomolecules and the hydrophobicity/hydrophilicity of the polymeric substrate on selective adhesion of endothelial cells (ECs, fibroblasts (FBs, and smooth muscle cells (SMCs. To control the polymeric substrate, biomolecules are immobilized on thermoresponsive poly(N-isopropylacrylamide-co-2-carboxyisopropylacrylamide (poly(NIPAAm-co-CIPAAm-grafted glass surfaces. By switching the molecular conformation of the biomolecule-immobilized polymers, the cell-selective adhesion performances are evaluated. In case of RGDS (Arg-Gly-Asp-Ser peptide-immobilized surfaces, all cell types adhere well regardless of the surface hydrophobicity. On the other hand, a tri-Arg-immobilized surface exhibits FB-selectivity when the surface is hydrophilic. Additionally, a tri-Ile-immobilized surface exhibits EC-selective cell adhesion when the surface is hydrophobic. We believe that the proposed concept, which is used to investigate the biomolecule-immobilized surface combination, is important to produce new biomaterials, which are highly demanded for medical implants and tissue engineering.

  16. Cell Adhesion on Surface-Functionalized Magnesium.

    Science.gov (United States)

    Wagener, Victoria; Schilling, Achim; Mainka, Astrid; Hennig, Diana; Gerum, Richard; Kelch, Marie-Luise; Keim, Simon; Fabry, Ben; Virtanen, Sannakaisa

    2016-05-18

    The biocompatibility of commercially pure magnesium-based (cp Mg) biodegradable implants is compromised of strong hydrogen evolution and surface alkalization due to high initial corrosion rates of cp Mg in the physiological environment. To mitigate this problem, the addition of corrosion-retarding alloying elements or coating of implant surfaces has been suggested. In the following work, we explored the effect of organic coatings on long-term cell growth. cp Mg was coated with aminopropyltriehtoxysilane + vitamin C (AV), carbonyldiimidazole (CDI), or stearic acid (SA). All three coatings have been previously suggested to reduce initial corrosion and to enhance protein adsorption and hence cell adhesion on magnesium surfaces. Endothelial cells (DH1+/+) and osteosarcoma cells (MG63) were cultured on coated samples for up to 20 days. To quantify Mg corrosion, electrochemical impedance spectroscopy (EIS) was measured after 1, 3, and 5 days of cell culture. We also investigated the speed of initial cell spreading after seeding using fluorescently labeled fibroblasts (NIH/3T3). Hydrogen evolution after contact with cell culture medium was markedly decreased on AV- and SA-coated Mg compared to uncoated Mg. These coatings also showed improved cell adhesion and spreading after 24 h of culture comparable to tissue-treated plastic surfaces. On AV-coated cp Mg, a confluent layer of endothelial cells formed after 5 days and remained intact for up to 20 days. Together, these data demonstrate that surface coating with AV is a viable strategy for improving long-term biocompatibility of cp Mg-based implants. EIS measurements confirmed that the presence of a confluent cell layer increased the corrosion resistance. PMID:27089250

  17. Cell surface proteome of the marine planctomycete Rhodopirellula baltica.

    Science.gov (United States)

    Voigt, Birgit; Hieu, Cao Xuan; Hempel, Kristina; Becher, Dörte; Schlüter, Rabea; Teeling, Hanno; Glöckner, Frank Oliver; Amann, Rudolf; Hecker, Michael; Schweder, Thomas

    2012-06-01

    The surface proteome (surfaceome) of the marine planctomycete Rhodopirellula baltica SH1(T) was studied using a biotinylation and a proteinase K approach combined with SDS-PAGE and mass spectrometry. 52 of the proteins identified in both approaches could be assigned to the group of potential surface proteins. Among them are some high molecular weight proteins, potentially involved in cell-cell attachment, that contain domains shown before to be typical for surface proteins like cadherin/dockerin domains, a bacterial adhesion domain or the fasciclin domain. The identification of proteins with enzymatic functions in the R. baltica surfaceome provides further clues for the suggestion that some degradative enzymes may be anchored onto the cell surface. YTV proteins, which have been earlier supposed to be components of the proteinaceous cell wall of R. baltica, were detected in the surface proteome. Additionally, 8 proteins with a novel protein structure combining a conserved type IV pilin/N-methylation domain and a planctomycete-typical DUF1559 domain were identified. PMID:22623273

  18. Biosensing based on surface plasmon resonance and living cells.

    Science.gov (United States)

    Chabot, Vincent; Cuerrier, Charles M; Escher, Emanuel; Aimez, Vincent; Grandbois, Michel; Charette, Paul G

    2009-02-15

    We propose the combination of surface plasmon resonance (SPR) with living cells as a biosensing method. Our detection scheme is based on the premise that cellular activity induced by external agents is often associated with changes in cellular morphology, which in turn should lead to a variation of the effective refractive index at the interface between the cell membrane and the metal layer. We monitored surface plasmon resonance signals originating from a gold surface coated with cells on a custom apparatus after injection of various agents known to influence cellular activity and morphology. Specifically, we evaluated three types of stimulation: response to an endotoxin (lipopolysaccharides), a chemical toxin (sodium azide) and a physiological agonist (thrombin). A comparison with phase contrast microscopy reveals that SPR signal variations are associated with the induction of cell death for lipopolysaccharides treatment and a contraction of the cell body for sodium azide. Thrombin-induced cellular response shows a rapid decrease of the measured laser reflectance over 5min followed by a return to the original value. For this treatment, phase contrast micrographs relate the first phase of the SPR variation to cell contraction and increase of the intercellular gaps, whereas the recovery phase can be associated with a spreading of the cell on the sensing surface. Hence, the SPR signal is very consistent with the cellular response normally observed for these treatments. This confirms the validity of the biosensing method, which could be applied to a large variety of cellular responses involving shape remodeling induced by external agents. PMID:18845432

  19. Surface cell immobilization within perfluoroalkoxy microchannels

    Energy Technology Data Exchange (ETDEWEB)

    Stojkovič, Gorazd; Krivec, Matic [Faculty of Chemistry and Chemical Technology, University of Ljubljana, Aškerčeva 5, SI-1000 Ljubljana (Slovenia); Vesel, Alenka [Jožef Stefan Institute, Jamova cesta 39, 1000 Ljubljana (Slovenia); Marinšek, Marjan [Faculty of Chemistry and Chemical Technology, University of Ljubljana, Aškerčeva 5, SI-1000 Ljubljana (Slovenia); Žnidaršič-Plazl, Polona, E-mail: polona.znidarsic@fkkt.uni-lj.si [Faculty of Chemistry and Chemical Technology, University of Ljubljana, Aškerčeva 5, SI-1000 Ljubljana (Slovenia)

    2014-11-30

    Graphical abstract: - Highlights: • A very efficient approach for immobilization of cells into microreactors is presented. • It is applicable to various materials, including PFA and cyclic olefin (co)polymers. • It was used to immobilize different prokaryotic and eukaryotic microbes. • Cells were immobilized on the surface in high density and showed good stability. • Mechanisms of APTES interactions with target materials are proposed. - Abstract: Perfluoroalkoxy (PFA) is one of the most promising materials for the fabrication of cheap, solvent resistant and reusable microfluidic chips, which have been recently recognized as effective tools for biocatalytic process development. The application of biocatalysts significantly depends on efficient immobilization of enzymes or cells within the reactor enabling long-term biocatalyst use. Functionalization of PFA microchannels by 3-aminopropyltriethoxysilane (ATPES) and glutaraldehyde was used for rapid preparation of microbioreactors with surface-immobilized cells. X-ray photoelectron spectroscopy and scanning electron microscopy were used to accurately monitor individual treatment steps and to select conditions for cell immobilization. The optimized protocol for Saccharomyces cerevisiae immobilization on PFA microchannel walls comprised ethanol surface pretreatment, 4 h contacting with 10% APTES aqueous solution, 10 min treatment with 1% glutaraldehyde and 20 min contacting with cells in deionized water. The same protocol enabled also immobilization of Escherichia coli, Pseudomonas putida and Bacillus subtilis cells on PFA surface in high densities. Furthermore, the developed procedure has been proved to be very efficient also for surface immobilization of tested cells on other materials that are used for microreactor fabrication, including glass, polystyrene, poly (methyl methacrylate), polycarbonate, and two olefin-based polymers, namely Zeonor{sup ®} and Topas{sup ®}.

  20. The cell surface proteome of human mesenchymal stromal cells.

    Directory of Open Access Journals (Sweden)

    Christian Niehage

    Full Text Available BACKGROUND: Multipotent human mesenchymal stromal cells (hMSCs are considered as promising biological tools for regenerative medicine. Their antibody-based isolation relies on the identification of reliable cell surface markers. METHODOLOGY/PRINCIPAL FINDINGS: To obtain a comprehensive view of the cell surface proteome of bone marrow-derived hMSCs, we have developed an analytical pipeline relying on cell surface biotinylation of intact cells using cell impermeable, cleavable sulfo-NHS-SS-biotin to enrich the plasma membrane proteins and mass spectrometry for identification with extremely high confidence. Among the 888 proteins identified, we found ≈200 bona fide plasma membrane proteins including 33 cell adhesion molecules and 26 signaling receptors. In total 41 CD markers including 5 novel ones (CD97, CD112, CD239, CD276, and CD316 were identified. The CD markers are distributed homogenously within plastic-adherent hMSC populations and their expression is modulated during the process of adipogenesis or osteogenesis. Moreover, our in silico analysis revealed a significant difference between the cell surface proteome of hMSCs and that of human embryonic stem cells reported previously. CONCLUSIONS/SIGNIFICANCE: Collectively, our analytical methods not only provide a basis for further studies of mechanisms maintaining the multipotency of hMSCs within their niches and triggering their differentiation after signaling, but also a toolbox for a refined antibody-based identification of hMSC populations from different tissues and their isolation for therapeutic intervention.

  1. An electrochemical surface plasmon resonance imaging system targeting cell analysis

    Science.gov (United States)

    Zhang, L. L.; Chen, X.; Wei, H. T.; Li, H.; Sun, J. H.; Cai, H. Y.; Chen, J. L.; Cui, D. F.

    2013-08-01

    This paper presents an electrochemical-surface plasmon resonance imaging (EC-SPRI) system, enabling the characterization of optical and electrical properties of cells, simultaneously. The developed surface plasmon resonance (SPR) imaging system was capable of imaging micro cavities with a dimension of 10 μm × 10 μm and differentiated glycerol solutions with a group of refractive indices (RIs). Furthermore, the EC-SPRI system was used to image A549 cells, suggesting corresponding RI and morphology changes during the cell death process. In the end, electrochemical and SPR methods were used in combination, recording oxidation peaks of A549 cells in the cyclic voltage curves and SPR response unit increase, simultaneously.

  2. Cell patterning on polylactic acid through surface-tethered oligonucleotides.

    Science.gov (United States)

    Matsui, Toshiki; Arima, Yusuke; Takemoto, Naohiro; Iwata, Hiroo

    2015-02-01

    Polylactic acid (PLA) is a candidate material to prepare scaffolds for 3-D tissue regeneration. However, cells do not adhere or proliferate well on the surface of PLA because it is hydrophobic. We report a simple and rapid method for inducing cell adhesion to PLA through DNA hybridization. Single-stranded DNA (ssDNA) conjugated to poly(ethylene glycol) (PEG) and to a terminal phospholipid (ssDNA-PEG-lipid) was used for cell surface modification. Through DNA hybridization, modified cells were able to attach to PLA surfaces modified with complementary sequence (ssDNA'). Different cell types can be attached to PLA fibers and films in a spatially controlled manner by using ssDNAs with different sequences. In addition, they proliferate well in a culture medium supplemented with fetal bovine serum. The coexisting modes of cell adhesion through DNA hybridization and natural cytoskeletal adhesion machinery revealed no serious effects on cell growth. The combination of a 3-D scaffold made of PLA and cell immobilization on the PLA scaffold through DNA hybridization will be useful for the preparation of 3-D tissue and organs.

  3. 30 CFR 785.18 - Variances for delay in contemporaneous reclamation requirement in combined surface and...

    Science.gov (United States)

    2010-07-01

    ... reclamation requirement in combined surface and underground mining activities. 785.18 Section 785.18 Mineral... reclamation requirement in combined surface and underground mining activities. (a) Scope. This section shall apply to any person or persons conducting or intending to conduct combined surface and...

  4. Effect of a combined inversion and plantarflexion surface on ankle kinematics and EMG activities in landing

    Directory of Open Access Journals (Sweden)

    Divya Bhaskaran

    2015-12-01

    Conclusion: These findings suggest that compared to the inversion surface, the combined plantarflexion and inversion surface seems to provide a more unstable surface condition for lateral ankle sprains during landing.

  5. Combined surface acoustic wave and surface plasmon resonance measurement of collagen and fibrinogen layers

    CERN Document Server

    Friedt, J M; Francis, L; Zhou, C; Campitelli, A; Friedt, Jean-Michel; Denis, Frederic; Francis, Laurent; Zhou, Cheng; Campitelli, Andrew

    2003-01-01

    We use an instrument combining optical (surface plasmon resonance) and acoustic (Love mode acoustic wave device) real-time measurements on a same surface for the identification of water content in collagen and fibrinogen protein layers. After calibration of the surface acoustic wave device sensitivity by copper electrodeposition, the bound mass and its physical properties -- density and optical index -- are extracted from the complementary measurement techniques and lead to thickness and water ratio values compatible with the observed signal shifts. Such results are especially usefully for protein layers with a high water content as shown here for collagen on an hydrophobic surface. We obtain the following results: collagen layers include 70+/-20 % water and are 16+/-3 to 19+/-3 nm thick for bulk concentrations ranging from 30 to 300 ug/ml. Fibrinogen layers include 50+/-10 % water for layer thicknesses in the 6+/-1.5 to 13+/-2 nm range when the bulk concentration is in the 46 to 460 ug/ml range.

  6. Model heterogeneous acid catalysts and metal-support interactions: A combined surface science and catalysis study

    Energy Technology Data Exchange (ETDEWEB)

    Boszormenyi, I.

    1991-05-01

    This (<100 {Angstrom}) silica-alumina layers were tested as potential model heterogeneous acid catalysts for combined surface science and catalysis studies. Three preparation methods were used: oxidation of r3 {times} r3 R30 Al/Si(111) structure in UHV; deposition on Si(lll) from aqueous solution; and argon ion beam sputter deposition in UHV. The homogeneous thin layers are amorphous, and the chemical environment of surface atoms is similar to that of Si, Al and oxygen atoms on high surface area acid catalysts. Since the ion beam-deposited thin layer of silica-alumina has the same composition as the target zeolite this deposition method is a promising tool to prepare model catalysts using practical catalyst targets. The silica-alumina layers are active in cumene cracking, a typical acid catalyzed reaction. In order to clearly distinguish background reactions and the acid catalyzed reaction at least 20 cm{sup 2} catalyst surface area is needed. Two series of model platinum-alumina catalysts were prepared in a combined UHV -- high pressure reactor cell apparatus by depositing alumina on polycrystalline Pt foil and by vapor depositing Pt on a thin alumina layer on Au. Both model surfaces have been prepared with and without chlorine. AES, CO desorption as well as methyl cyclopentane (MCP) hydrogenolysis studies indicate that the Pt surface area is always higher if a chlorination step is involved. Selectivity patterns in MCP ring opening on ``Pt-on-alumina`` and on ``alumina-on-Pt`` are different; only the former is a linear combination of selective and statistical ring opening. Product distribution, however, changes with coverage and reaction time. The properties of the two model catalyst systems and role of chlorine in MCP hydrogenolysis are also discussed.

  7. Model heterogeneous acid catalysts and metal-support interactions: A combined surface science and catalysis study

    Energy Technology Data Exchange (ETDEWEB)

    Boszormenyi, I.

    1991-05-01

    This (<100 [Angstrom]) silica-alumina layers were tested as potential model heterogeneous acid catalysts for combined surface science and catalysis studies. Three preparation methods were used: oxidation of r3 [times] r3 R30 Al/Si(111) structure in UHV; deposition on Si(lll) from aqueous solution; and argon ion beam sputter deposition in UHV. The homogeneous thin layers are amorphous, and the chemical environment of surface atoms is similar to that of Si, Al and oxygen atoms on high surface area acid catalysts. Since the ion beam-deposited thin layer of silica-alumina has the same composition as the target zeolite this deposition method is a promising tool to prepare model catalysts using practical catalyst targets. The silica-alumina layers are active in cumene cracking, a typical acid catalyzed reaction. In order to clearly distinguish background reactions and the acid catalyzed reaction at least 20 cm[sup 2] catalyst surface area is needed. Two series of model platinum-alumina catalysts were prepared in a combined UHV -- high pressure reactor cell apparatus by depositing alumina on polycrystalline Pt foil and by vapor depositing Pt on a thin alumina layer on Au. Both model surfaces have been prepared with and without chlorine. AES, CO desorption as well as methyl cyclopentane (MCP) hydrogenolysis studies indicate that the Pt surface area is always higher if a chlorination step is involved. Selectivity patterns in MCP ring opening on Pt-on-alumina'' and on alumina-on-Pt'' are different; only the former is a linear combination of selective and statistical ring opening. Product distribution, however, changes with coverage and reaction time. The properties of the two model catalyst systems and role of chlorine in MCP hydrogenolysis are also discussed.

  8. Femtosecond fabricated surfaces for cell biology

    Science.gov (United States)

    Day, Daniel; Gu, Min

    2010-08-01

    Microfabrication using femtosecond pulse lasers is enabling access to a range of structures, surfaces and materials that was not previously available for scientific and engineering applications. The ability to produce micrometre sized features directly in polymer and metal substrates is demonstrated with applications in cell biology. The size, shape and aspect ratio of the etched features can be precisely controlled through the manipulation of the fluence of the laser etching process with respect to the properties of the target material. Femtosecond laser etching of poly(methyl methacrylate) and aluminium substrates has enabled the production of micrometre resolution moulds that can be accurately replicated using soft lithography. The moulded surfaces are used in the imaging of T cells and demonstrate the improved ability to observe biological events over time periods greater than 10 h. These results indicate the great potential femtosecond pulse lasers may have in the future manufacturing of microstructured surfaces and devices.

  9. Nanofiber-modified surface directed cell migration and orientation in microsystem

    Science.gov (United States)

    Zhang, Xu; Gao, Xinghua; Jiang, Lei; Zhang, Xulang; Qin, Jianhua

    2011-01-01

    Cell-microscale pattern surface interactions are crucial to understand many fundamental biological questions and develop regenerative medicine and tissue engineering approaches. In this work, we demonstrated a simple method to pattern PDMS surface by sacrificing poly vinyl pyrrolidone (PVP) electrospinning nanofibers and investigated the growth profile of cells on the modified patterned surfaces using stroma cells. The stromal cells were observed to exhibit good viability on this modified surface and the patterned surface with alignment nanofibers could promote cell migration. Furthermore, the modified PDMS surface was integrated with microfluidic channels to create the microscale spatial factor and was used to explore the cell migration and orientation under this microsystem. Both spatial factor and patterned surfaces were found to contribute to the complex cell orientation under the combined dual effects. This established method is simple, fast, and easy for use, demonstrating the potential of this microsystem for applications in addressing biological questions in complex environment. PMID:22662030

  10. Engineered microtopographies and surface chemistries direct cell attachment and function

    Science.gov (United States)

    Magin, Chelsea Marie

    topographically modified surface (R2=0.82). Functionalized PEGDMA hydrogels significantly reduced attachment and attachment strength of Navicula and C. marina. These hydrogels also reduced attachment of zoospores of Ulva compared to PDMSe. Attachment of Ulva to microtopographies in PDMSe and PEGDMA-co-HEMA negatively correlated with ERIII*Re (R2 = 0.94 and R2 = 0.99, respectively). Incorporating a surface energy term into this equation created a correlation between the attachment densities of cells from two evolutionarily diverse groups on substrates of two surface chemistries with an equation that describes the various microtopographies and surface chemistries in terms of surface energy (R2 = 0.80). The current Attachment Model can now be used to design engineered antifouling surface microtopographies and chemistries that inhibit the attachment of organisms from three evoluntionarily diverse groups. Hydrogels based on PEGDMA were also chosen as a substratum material for mammalian cell culture. Capturing endothelial progenitor cells (EPCs) and inducing differentiation into the endothelial cell (EC) phenotype is the ideal way to re-endothelialize a small-diameter vascular graft. Substratum elasticity has been reported to direct stem cell differentiation into specific lineages. Functionalized PEGDMA hydrogels provided good compliance, high fidelity of topographic features and sites for surface modification with biomolecules. Fibronectin grafting and topography both increased EC attachment. This combination of adjustable elasticity, surface chemistry and topography has the potential to promote the capture and differentiation of EPCs into a confluent EC monolayer. Engineered microtopographies replicated in PDMSe directed elongation and alignment of human coronary artery endothelial cells (HCAECs) and human coronary artery smooth muscle cells (HCASMCs) compared to smooth surfaces. Engineered cellular micro-environments were created with specific surface energies defined by chemistry

  11. Impurity concentrations and surface charge densities on the heavily doped face of a silicon solar cell

    Science.gov (United States)

    Weinberg, I.; Hsu, L. C.

    1977-01-01

    Increased solar cell efficiencies are attained by reduction of surface recombination and variation of impurity concentration profiles at the n(+) surface of silicon solar cells. Diagnostic techniques are employed to evaluate the effects of specific materials preparation methodologies on surface and near surface concentrations. It is demonstrated that the MOS C-V method, when combined with a bulk measurement technique, yields more complete concentration data than are obtainable by either method alone. Specifically, new solar cell MOS C-V measurements are combined with bulk concentrations obtained by a successive layer removal technique utilizing measurements of sheet resistivity and Hall coefficient.

  12. SURFACE CAST IRON STRENGTHENING USING COMBINED LASER AND ULTRASONIC PROCESSING

    Directory of Open Access Journals (Sweden)

    O. Devojno

    2013-01-01

    Full Text Available The paper provides an analysis of ultrasonic surface plastic deformation and subsequent laser thermal strengthening of gray cast iron parts in the regime of hardening from a solid state with the purpose to obtain strengthened surface layers of bigger depth and less roughness of the processed surface. Program complex ANSYS 11.0 has been used for calculation of temperature fields induced by laser exposure.  The appropriate regime of laser processing without surface fusion has been selected on the basis of the applied complex. The possibility of displacement in the bottom boundary of α–γ-transformation temperature  for СЧ20 with 900 °С up to 800 °С is confirmed due to preliminary ultrasonic surface plastic deformation of the surface that allows to expand technological opportunities of laser quenching  of gray  cast iron from a solid state. 

  13. A new approach to study local corrosion processes on steel surfaces by combining different microscopic techniques

    Science.gov (United States)

    Heyer, A.; D'Souza, F.; Bruin, A.; Ferrari, G.; Mol, J. M. C.; de Wit, J. H. W.

    2012-09-01

    Corrosion studies of materials on the micro or even nano-scale level are cumbersome due to instrumental limitations and handling procedures. If biological processes are involved the spatial resolution is even more important and sample preparation is usually the limitation. Attachment of bacteria on stainless steel surface is a complex interfacial process including interactions of bacterial cells and bacterial extracellular polymeric substances with the surface. To overcome the limitations in sample preparations and resolution we present a new stainless steel sample holder to switch among epifluorescent microscope (EFM), AFM and SEM at exactly the same position. Exemplary bacterial accumulation was studied by staining the bacterial DNA with a fluorescent dye over time. It was possible to distinguish among bacteria and other surface characteristic such as deformations or grain structures. Also surface topographic features such as roughness at the grain boundaries and deposits were quantified. All three techniques complement one another in the way that AFM is a high-resolution technique that does not allow to distinguish directly bacterial cell structures, whereas EFM offers excellent bacterial identification based on staining at a low resolution that can complement AFM images. Application of SEM in the last step will reveal inclusions and grain structure and combined with EDX gives the composition of the substrate, inclusions and corrosion deposit. The combination of the three high-resolution techniques enables a more detailed understanding of surface phenomena. The method itself is quite elegant and easy to handle which is an important aspect in materials research, especially when a high sample throughput is needed.

  14. Engineering novel cell surface chemistry for selective tumor cell targeting

    Energy Technology Data Exchange (ETDEWEB)

    Bertozzi, C.R. [Univ. of California, Berkeley, CA (United States)]|[Lawrence Berkeley National Lab., CA (United States)

    1997-12-31

    A common feature of many different cancers is the high expression level of the two monosaccharides sialic acid and fucose within the context of cell-surface associated glycoconjugates. A correlation has been made between hypersialylation and/or hyperfucosylation and the highly metastatic phenotype. Thus, a targeting strategy based on sialic acid or fucose expression would be a powerful tool for the development of new cancer cell-selective therapies and diagnostic agents. We have discovered that ketone groups can be incorporated metabolically into cell-surface associated sialic acids. The ketone is can be covalently ligated with hydrazide functionalized proteins or small molecules under physiological conditions. Thus, we have discovered a mechanism to selectively target hydrazide conjugates to highly sialylated cells such as cancer cells. Applications of this technology to the generation of novel cancer cell-selective toxins and MRI contrast reagents will be discussed, in addition to progress towards the use of cell surface fucose residues as vehicles for ketone expression.

  15. Emergence of an Apical Epithelial Cell Surface In Vivo.

    Science.gov (United States)

    Sedzinski, Jakub; Hannezo, Edouard; Tu, Fan; Biro, Maté; Wallingford, John B

    2016-01-11

    Epithelial sheets are crucial components of all metazoan animals, enclosing organs and protecting the animal from its environment. Epithelial homeostasis poses unique challenges, as addition of new cells and loss of old cells must be achieved without disrupting the fluid-tight barrier and apicobasal polarity of the epithelium. Several studies have identified cell biological mechanisms underlying extrusion of cells from epithelia, but far less is known of the converse mechanism by which new cells are added. Here, we combine molecular, pharmacological, and laser-dissection experiments with theoretical modeling to characterize forces driving emergence of an apical surface as single nascent cells are added to a vertebrate epithelium in vivo. We find that this process involves the interplay between cell-autonomous actin-generated pushing forces in the emerging cell and mechanical properties of neighboring cells. Our findings define the forces driving this cell behavior, contributing to a more comprehensive understanding of epithelial homeostasis. PMID:26766441

  16. A combination of CO{sub 2} laser and plasma surface modification of poly(etheretherketone) to enhance osteoblast response

    Energy Technology Data Exchange (ETDEWEB)

    Zheng, Yanyan [Chengdu Institute of Organic Chemistry, Chinese Academy of Sciences, Chengdu 610041 (China); University of Chinese Academy of Sciences, Beijing 100049 (China); Xiong, Chengdong [Chengdu Institute of Organic Chemistry, Chinese Academy of Sciences, Chengdu 610041 (China); Wang, Zhecun [Chengdu Institute of Organic Chemistry, Chinese Academy of Sciences, Chengdu 610041 (China); University of Chinese Academy of Sciences, Beijing 100049 (China); Li, Xiaoyu [State Key Laboratory of Oral Diseases, West China Hospital of Stomatology, Sichuan University, Chengdu 610041 (China); Zhang, Lifang, E-mail: zhanglfcioc@163.com [Chengdu Institute of Organic Chemistry, Chinese Academy of Sciences, Chengdu 610041 (China)

    2015-07-30

    Highlights: • COOH and microgrooves containing micropores or microcraters structure were constructed on PEEK surface by a combination of CO{sub 2} laser and plasma treatment. • The mechanical properties of PEEK are maintained after single or dual surface treatment. • Pre-osteoblast cells (MC3T3-E1) adhesion, spreading and proliferation were improved remarkably on dual treated PEEK surface. • Cell pseudopodia protrude into the micropores or microcraters, in favor of forming firmer bone-implant integration. - Abstract: Poly(etheretherketone) (PEEK) is a rigid semicrystalline polymer that combines excellent mechanical properties, broad chemical resistance and bone-like stiffness and is widely used in biomedical fields. However, the bio-inert surface of PEEK tends to hinder its biomedical applications when direct osteointegration between the implants and the host tissue is desired. In this work, we demonstrate a dual modification method, which combines the laser and plasma surface treatment to combine advantages of both chemical states and microstructures for osteoblasts responses. While the plasma treatment introduces surface carboxyl groups (−COOH) onto PEEK surface, the laser treatment constructs microstructures over the PEEK surface. Our results indicated that −COOH as well as microgrooves containing micropores or microcraters structure are constructed on PEEK surface and plasma treatment has no apparent effect on the morphology of microstructures produced by laser micromachining. Unexpectedly, the superior mechanical properties of PEEK were maintained irrespective of the treatment used. Compared to native PEEK and single treated PEEK, dual modified PEEK is more favorable for pre-osteoblasts (MC3T3-E1) adhesion, spreading and proliferation. Moreover, cell pseudopodia protrude into the micropores or microcraters, in favor of forming firmer bone-implant integration. Our study illustrates enhanced osteoblasts responses to dual treated PEEK surface, which

  17. Effect of mutagen combined action on Chlamydomonas reinhardtii cells. II

    International Nuclear Information System (INIS)

    The effect of UV radiation and UV radiation combined with alkylnitrosourea derivatives (N-methyl-N-nitrosourea and N-ethyl-N-nitrosourea) was observed on survival of cells of the algae Chlamydomonas reinhardtii. In particular, single parts were evaluated of the overall lethal effect - dying of cells before division and dying of cells after division. It was found that the combined action of low doses of UV radiation and alkylnitrosoureas result in a pronounced protective effect which manifests itself by a higher frequency of surviving cells than was that effected by the action of alkylnitrosoureas alone. As a result of combined action with higher doses of UV radiation this effect is lost, and the resultant values will come close to the theoretically anticipated values. This gradual transition from a protective to an additive effect mainly manifests itself by changes in the proportion of cells dying before division. (author)

  18. Durable Microstructured Surfaces: Combining Electrical Conductivity with Superoleophobicity.

    Science.gov (United States)

    Pan, Zihe; Wang, Tianchang; Sun, Shaofan; Zhao, Boxin

    2016-01-27

    In this study, electrically conductive and superoleophobic polydimethylsiloxane (PDMS) has been fabricated through embedding Ag flakes (SFs) and Ag nanowires (SNWs) into microstructures of the trichloroperfluorooctylsilane (FDTS)-blended PDMS elastomer. Microstructured PDMS surfaces became conductive at the percolation surface coverage of 3.0 × 10(-2) mg/mm(2) for SFs; the highest conductivity was 1.12 × 10(5) S/m at the SFs surface coverage of 6.0 × 10(-2) mg/mm(2). A significant improvement of the conductivity (increased 3 times at the SNWs fraction of 11%) was achieved by using SNWs to replace some SFs because of the conductive pathways from the formed SNWs networks and its connections with SFs. These conductive fillers bonded strongly with microstructured FDTS-blended PDMS and retained surface properties under the sliding preload of 8.0 N. Stretching tests indicated that the resistance increased with the increasing strains and returned to its original state when the strain was released, showing highly stretchable and reversible electrical properties. Compared with SFs embedded surfaces, the resistances of SFs/SNWs embedded surfaces were less dependent on the strain because of bridging effect of SNWs. The superoleophobicity was achieved by the synergetic effect of surface modification through blending FDTS and the microstructures transferred from sand papers. The research findings demonstrate a simple approach to make the insulating elastomer to have the desired surface oleophobicity and electrical conductivity and help meet the needs for the development of conductive devices with microstructures and multifunctional properties.

  19. Morphological investigations of cells that adhered to the irregular patterned polydimethylsiloxane (PDMS) surface without reagents.

    Science.gov (United States)

    Chung, Sung Hee; Min, Junhong

    2009-07-01

    Polydimethylsiloxane (PDMS) surface consisting irregular pattern was investigated to develop cell-based biochip using PDMS. PDMS surface was modified with nano- and micro-combined patterns using surface deformation technology. Hydrophobicity of nano-patterned PDMS surface was sustained. Nevertheless it has irregular patterns consisting of micro- and nano-patterns. According to atomic force microscopy (AFM), scanning electron microscopy (SEM) and confocal microscopy results by immunostaining method, human mammary epithelial cells (HMEC) adhered well on irregularly patterned surface without any reagents such as gelatin and collagen, compared to commercial culture dish. It implies PDMS material can be utilized as template for cell-based biochip without any reagents. PMID:19427124

  20. Robust combined position and formation control for marine surface craft

    DEFF Research Database (Denmark)

    Ihle, Ivar-Andre F.; Jouffroy, Jerome; Fossen, Thor I.

    We consider the robustness properties of a formation control system for marine surface vessels. Intervessel constraint functions are stabilized to achieve the desired formation configuration. We show that the formation dynamics is Input-to-State Stable (ISS) to both environmental perturbations th...

  1. Surface modified alginate microcapsules for 3D cell culture

    Science.gov (United States)

    Chen, Yi-Wen; Kuo, Chiung Wen; Chueh, Di-Yen; Chen, Peilin

    2016-06-01

    Culture as three dimensional cell aggregates or spheroids can offer an ideal platform for tissue engineering applications and for pharmaceutical screening. Such 3D culture models, however, may suffer from the problems such as immune response and ineffective and cumbersome culture. This paper describes a simple method for producing microcapsules with alginate cores and a thin shell of poly(L-lysine)-graft-poly(ethylene glycol) (PLL-g-PEG) to encapsulate mouse induced pluripotent stem (miPS) cells, generating a non-fouling surface as an effective immunoisolation barrier. We demonstrated the trapping of the alginate microcapsules in a microwell array for the continuous observation and culture of a large number of encapsulated miPS cells in parallel. miPS cells cultured in the microcapsules survived well and proliferated to form a single cell aggregate. Droplet formation of monodisperse microcapsules with controlled size combined with flow cytometry provided an efficient way to quantitatively analyze the growth of encapsulated cells in a high-throughput manner. The simple and cost-effective coating technique employed to produce the core-shell microcapsules could be used in the emerging field of cell therapy. The microwell array would provide a convenient, user friendly and high-throughput platform for long-term cell culture and monitoring.

  2. CZTSSe thin film solar cells: Surface treatments

    Science.gov (United States)

    Joglekar, Chinmay Sunil

    Chalcopyrite semiconducting materials, specifically CZTS, are a promising alternative to traditional silicon solar cell technology. Because of the high absorption coefficient; films of the order of 1 micrometer thickness are sufficient for the fabrication of solar cells. Liquid based synthesis methods are advantageous because they are easily scalable using the roll to roll manufacturing techniques. Various treatments are explored in this study to enhance the performance of the selenized CZTS film based solar cells. Thiourea can be used as a sulfur source and can be used to tune band gap of CZTSSe. Bromine etching can be used to manipulate the thickness of sintered CZTSSe film. The etching treatment creates recombination centers which lead to poor device performance. Various after treatments were used to improve the performance of the devices. It was observed that the performance of the solar cell devices could not be improved by any of the after treatment steps. Other surface treatment processes are explored including KCN etching and gaseous H2S treatments. Hybrid solar cells which included use of CIGS nanoparticles at the interface between CZTSSe and CdS are also explored.

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

    Directory of Open Access Journals (Sweden)

    Tohru Hayakawa

    2012-01-01

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

  4. Combination of fluid and solid mechanical stresses contribute to cell death and detachment in a microfluidic alveolar model.

    Science.gov (United States)

    Douville, Nicholas J; Zamankhan, Parsa; Tung, Yi-Chung; Li, Ran; Vaughan, Benjamin L; Tai, Cheng-Feng; White, Joshua; Christensen, Paul J; Grotberg, James B; Takayama, Shuichi

    2011-02-21

    Studies using this micro-system demonstrated significant morphological differences between alveolar epithelial cells (transformed human alveolar epithelial cell line, A549 and primary murine alveolar epithelial cells, AECs) exposed to combination of solid mechanical and surface-tension stresses (cyclic propagation of air-liquid interface and wall stretch) compared to cell populations exposed solely to cyclic stretch. We have also measured significant differences in both cell death and cell detachment rates in cell monolayers experiencing combination of stresses. This research describes new tools for studying the combined effects of fluid mechanical and solid mechanical stress on alveolar cells. It also highlights the role that surface tension forces may play in the development of clinical pathology, especially under conditions of surfactant dysfunction. The results support the need for further research and improved understanding on techniques to reduce and eliminate fluid stresses in clinical settings. PMID:21152526

  5. Surface deformation and shear flow in ligand mediated cell adhesion

    Science.gov (United States)

    Sircar, Sarthok; Roberts, Anthony; Sarthok Sircar / Anthony Roberts Collaboration

    We present a unified, multiscale model to study the attachment/detachment dynamics of two deforming, near spherical cells, coated with binding ligands and subject to a slow, homogeneous shear flow in a viscous fluid medium. The binding ligands on the surface of the cells experience attractive and repulsive forces in an ionic medium and exhibit finite resistance to rotation via bond tilting. The microscale drag forces and couples describing the fluid flow inside the small separation gap between the cells, are calculated using a combination of methods in lubrication theory and previously published numerical results. For a select range of material and fluid parameters, a hysteretic transition of the sticking probability curves (i.e., the function g*) between the adhesion phase (when g*>0.5) and the fragmentation phase (when g*University startup funds and AR is supported by the Australian Research Council Discovery Grant DP150102385.

  6. Frequency Selective Surfaces with Nanoparticles Unit Cell

    Directory of Open Access Journals (Sweden)

    Nga Hung Poon

    2015-09-01

    Full Text Available The frequency selective surface (FSS is a periodic structure with filtering performance for optical and microwave signals. The general periodic arrays made with patterned metallic elements can act as an aperture or patch on a substrate. In this work, two kinds of materials were used to produce unit cells with various patterns. Gold nanoparticles of 25 nm diameter were used to form periodic monolayer arrays by a confined photocatalytic oxidation-based surface modification method. As the other material, silver gel was used to create multiple layers of silver. Due to the ultra-thin nature of the self-assembled gold nanoparticle monolayer, it is very easy to penetrate the FSS with terahertz radiation. However, the isolated silver islands made from silver gel form thicker multiple layers and contribute to much higher reflectance. This work demonstrated that multiple silver layers are more suitable than gold nanoparticles for use in the fabrication of FSS structures.

  7. Targeting pancreatic progenitor cells in human embryonic stem cell differentiation for the identification of novel cell surface markers.

    Science.gov (United States)

    Fishman, Bettina; Segev, Hanna; Kopper, Oded; Nissenbaum, Jonathan; Schulman, Margarita; Benvenisty, Nissim; Itskovitz-Eldor, Joseph; Kitsberg, Danny

    2012-09-01

    New sources of beta cells are needed in order to develop cell therapies for patients with diabetes. An alternative to forced expansion of post-mitotic beta cells is the induction of differentiation of stem-cell derived progenitor cells that have a natural self-expansion capacity into insulin-producing cells. In order to learn more about these progenitor cells at different stages along the differentiation process in which they become progressively more committed to the final beta cell fate, we took the approach of identifying, isolating and characterizing stage specific progenitor cells. We generated human embryonic stem cell (HESC) clones harboring BAC GFP reporter constructs of SOX17, a definitive endoderm marker, and PDX1, a pancreatic marker, and identified subpopulations of GFP expressing cells. Using this approach, we isolated a highly enriched population of pancreatic progenitor cells from hESCs and examined their gene expression with an emphasis on the expression of stage-specific cell surface markers. We were able to identify novel molecules that are involved in the pancreatic differentiation process, as well as stage-specific cell markers that may serve to define (alone or in combination with other markers) a specific pancreatic progenitor cell. These findings may help in optimizing conditions for ultimately generating and isolating beta cells for transplantation therapy.

  8. Combination Effect of Nimotuzumab with Radiation in Colorectal Cancer Cells

    Energy Technology Data Exchange (ETDEWEB)

    Shin, Hye Kyung; Kim, Mi Sook; Jeong, Jae Hoon [Korea Institute of Radiologicaland Medical Sciences, Seoul (Korea, Republic of)

    2010-11-15

    To investigate the radiosensitizing effect of the selective epidermal growth factor receptor (EGFR) inhibitor nimotuzumab in human colorectal cancer cell lines. Four human colorectal cancer cell lines, HCT-8, LoVo, WiDr, and HCT-116 were treated with nimotuzumab and/or radiation. The effects on cell proliferation, viability, and cell cycle progression were measured by MTT, clonogenic survival assay, flow cytometry, and Western blot. An immunoblot analysis revealed that EGFR phosphorylation was inhibited by nimotuzumab in colorectal cancer cell lines. Under these experimental conditions, pre-treatment with nimotuzumab increased radiosensitivity of colorectal cancer cell lines, except for cell line HCT-116. However, cell proliferation or cell cycle progression was not affected by the addition of nimotuzumab, irrespective of irradiation. Nimotuzumab enhanced the radiosensitivity of colorectal cancer cells in vitro by inhibiting EGFR-mediated cell survival signaling. This study provided a rationale for the clinical application of the selective EGFR inhibitor, nimotuzumab in combination with radiation in colorectal cancer cells.

  9. Production of alloyed nanocrystalline surfaces by combined punching, sandblasting and recovery treatments

    Energy Technology Data Exchange (ETDEWEB)

    Tang Xinhu [Department of Chemical and Materials Engineering, University of Alberta, Edmonton, T6G 2G6 (Canada); Li, D.Y. [Department of Chemical and Materials Engineering, University of Alberta, Edmonton, T6G 2G6 (Canada)], E-mail: dongyang.li@ualberta.ca

    2008-06-15

    The paper describes a process combining punching, sandblasting and recovery treatments (PSR) for synthesizing an alloyed Al-Ti nanocrystalline surface layer on an Al alloy. The treated surface was about five times harder than that of the original alloy and the surface stability increased markedly, as indicated by an increase in the electron work function. The corrosion resistance of the treated surface was also raised significantly. These changes originated from surface alloying and nanocrystallization.

  10. Near-surface alloys for hydrogen fuel cell applications

    DEFF Research Database (Denmark)

    Greeley, Jeffrey Philip; Mavrikakis, Manos

    2006-01-01

    Near-surface alloys (NSAs) possess a variety of unusual catalytic properties that could make them useful candidates for improved catalysts in a variety of chemical processes. It is known from previous work, for example, that some NSAs bind hydrogen very weakly while, at the same time, permitting...... facile H-2 activation. These NSAs could, potentially, facilitate highly selective hydrogenation reactions at low temperatures. In the present work, the suitability of NSAs for use as hydrogen fuel cell anodes has been evaluated: the combination of properties, possessed by selected NSAs, of weak binding...

  11. MUTUAL INHIBITION OF MURINE ERYTHROPOIESIS AND GRANULOPOIESIS DURING COMBINED ERYTHROPOIETIN, GRANULOCYTE-COLONY-STIMULATING FACTOR, AND STEM-CELL FACTOR ADMINISTRATION - IN-VIVO INTERACTIONS AND DOSE-RESPONSE SURFACES

    NARCIS (Netherlands)

    DEHAAN, G; ENGEL, C; DONTJE, B; NIJHOF, W; LOEFFLER, M

    1994-01-01

    We investigated the in vivo effects of erythropoietin (EPO) on granulopoiesis and, conversely, the effect of granulocyte colony-stimulating factor (G-CSF) treatment on erythropoiesis. Recombinant human EPO at four different doses in combination with recombinant human G-CSF also at four different dos

  12. Effect of mutagen combined action on Chlamydomonas Reinhardtii cells. I

    International Nuclear Information System (INIS)

    The effect was investigated of single and combined actions of alkylnitrosourea derivatives (N-methyl-N-nitrosourea and N-ethyl-N-nitrosourea) and UV-radiation on the survival of cells of Chlamydomonas reinhardtii algae in dependence on the sequence of application of mutagens and on the given conditions of cultivation following mutagen activity. In particular, the single phases were investigated of the total lethal effect, i.e., the death of cells before division and their death after division. The most pronounced changes in dependence on the sequence of application of mutagens and on the given conditions of cultivation were noted in cell death before division. In dependence on the sequence of application of mutagens, the effect of the combined action on the survival of cells changed from an additive (alkylnitrosourea + UV-radiation) to a protective effect (UV-radiation + alkylnitrosourea). (author)

  13. Cell receptor and surface ligand density effects on dynamic states of adhering circulating tumor cells.

    Science.gov (United States)

    Zheng, Xiangjun; Cheung, Luthur Siu-Lun; Schroeder, Joyce A; Jiang, Linan; Zohar, Yitshak

    2011-10-21

    Dynamic states of cancer cells moving under shear flow in an antibody-functionalized microchannel are investigated experimentally and theoretically. The cell motion is analyzed with the aid of a simplified physical model featuring a receptor-coated rigid sphere moving above a solid surface with immobilized ligands. The motion of the sphere is described by the Langevin equation accounting for the hydrodynamic loadings, gravitational force, receptor-ligand bindings, and thermal fluctuations; the receptor-ligand bonds are modeled as linear springs. Depending on the applied shear flow rate, three dynamic states of cell motion have been identified: (i) free motion, (ii) rolling adhesion, and (iii) firm adhesion. Of particular interest is the fraction of captured circulating tumor cells, defined as the capture ratio, via specific receptor-ligand bonds. The cell capture ratio decreases with increasing shear flow rate with a characteristic rate. Based on both experimental and theoretical results, the characteristic flow rate increases monotonically with increasing either cell-receptor or surface-ligand density within certain ranges. Utilizing it as a scaling parameter, flow-rate dependent capture ratios for various cell-surface combinations collapse onto a single curve described by an exponential formula.

  14. Surface recombination analysis in silicon-heterojunction solar cells

    Energy Technology Data Exchange (ETDEWEB)

    Barrio, R.; Gandia, J.J.; Carabe, J.; Gonzalez, N.; Torres, I. [CIEMAT, Madrid (Spain); Munoz, D.; Voz, C. [Universitat Politecnica de Catalunya, Barcelona (Spain)

    2010-02-15

    The origin of this work is the understanding of the correlation observed between efficiency and emitter-deposition temperature in single silicon-heterojunction solar cells prepared by depositing an n-doped hydrogenated-amorphous-silicon thin film onto a p-type crystalline-silicon wafer. In order to interpret these results, surface-recombination velocities have been determined by two methods, i.e. by fitting the current-voltage characteristics to a theoretical model and by means of the Quasi-Steady-State Photoconductance Technique (QSSPC). In addition, effective diffusion lengths have been estimated from internal quantum efficiencies. The analysis of these data has led to conclude that the performance of the cells studied is limited by back-surface recombination rather than by front-heterojunction quality. A 12%-efficient cell has been prepared by combining optimum emitter-deposition conditions with back-surface-field (BSF) formation by vacuum annealing of the back aluminium contact. This result has been achieved without using any transparent conductive oxide. (author)

  15. PROTEIN ADSORPTION AND CELL ADHESION ON RGD-FUNCTIONALIZED SILICON SUBSTRATE SURFACES

    Institute of Scientific and Technical Information of China (English)

    Wei-fang Tong; Xiao-li Liu; Fei Pan; Zhao-qiang Wu; Wen-wen Jiang

    2013-01-01

    A method was developed to modify silicon surfaces with good protein resistance and specific cell attachment.A silicon surface was initially deposited using a block copolymer of N-vinylpyrrolidone (NVP) and 2-hydroxyethyl methacrylate (HEMA) (PVP-b-PHEMA) film through surface-initiated atom transfer radical polymerization and then further immobilized using a short arginine-glycine-aspartate (RGD) peptide.Our results demonstrate that the RGD-modified surfaces (Si-RGD) can suppress non-specific adsorption of proteins and induce the adhesion of L929 cells.The Si-RGD surface exhibited higher cell proliferation rates than the unmodified silicon surface.This research established a simple method for the fabrication of dual-functional silicon surface that combines antifouling and cell attachment promotion.

  16. Manufacture of functional surfaces through combined application of tool manufacturing processes and Robot Assisted Polishing

    DEFF Research Database (Denmark)

    Eriksen, Rasmus Solmer; Arentoft, Mogens; Grønbæk, J.;

    2012-01-01

    The tool surface topography is often a key parameter in the tribological performance of modern metal forming tools. A new generation of multifunctional surfaces is achieved by combination of conventional tool manufacturing processes with a novel Robot Assisted Polishing process. This novel surface...

  17. Chemistry and material science at the cell surface

    Directory of Open Access Journals (Sweden)

    Weian Zhao

    2010-04-01

    Full Text Available Cell surfaces are fertile ground for chemists and material scientists to manipulate or augment cell functions and phenotypes. This not only helps to answer basic biology questions but also has diagnostic and therapeutic applications. In this review, we summarize the most recent advances in the engineering of the cell surface. In particular, we focus on the potential applications of surface engineered cells for 1 targeting cells to desirable sites in cell therapy, 2 programming assembly of cells for tissue engineering, 3 bioimaging and sensing, and ultimately 4 manipulating cell biology.

  18. WORKING PARK-FUEL CELL COMBINED HEAT AND POWER SYSTEM

    Energy Technology Data Exchange (ETDEWEB)

    Allan Jones

    2003-09-01

    This report covers the aims and objectives of the project which was to design, install and operate a fuel cell combined heat and power (CHP) system in Woking Park, the first fuel cell CHP system in the United Kingdom. The report also covers the benefits that were expected to accrue from the work in an understanding of the full technology procurement process (including planning, design, installation, operation and maintenance), the economic and environmental performance in comparison with both conventional UK fuel supply and conventional CHP and the commercial viability of fuel cell CHP energy supply in the new deregulated energy markets.

  19. Combining a Spatial Model and Demand Forecasts to Map Future Surface Coal Mining in Appalachia.

    Directory of Open Access Journals (Sweden)

    Michael P Strager

    Full Text Available Predicting the locations of future surface coal mining in Appalachia is challenging for a number of reasons. Economic and regulatory factors impact the coal mining industry and forecasts of future coal production do not specifically predict changes in location of future coal production. With the potential environmental impacts from surface coal mining, prediction of the location of future activity would be valuable to decision makers. The goal of this study was to provide a method for predicting future surface coal mining extents under changing economic and regulatory forecasts through the year 2035. This was accomplished by integrating a spatial model with production demand forecasts to predict (1 km2 gridded cell size land cover change. Combining these two inputs was possible with a ratio which linked coal extraction quantities to a unit area extent. The result was a spatial distribution of probabilities allocated over forecasted demand for the Appalachian region including northern, central, southern, and eastern Illinois coal regions. The results can be used to better plan for land use alterations and potential cumulative impacts.

  20. A combined electrochemical and DFT study of the lattice strain effect on the surface reactivity of Pd

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    We report a combined study of electrochemical experiments and ab initio calculations on tuning the surface reactivity of Pd via a compressive lattice strain achieved by employing nanoparticles of Pd-Cu alloys with a Pd-rich surface. Surface oxygen-containing species were used as the probing molecule for revealing the surface reactivity. Both density functional theory (DFT) calculations and experiments showed linear relationships, with very close slopes, between the adsorption strength of OHads and the Pd lattice constant. Not only is this work a successful realization of controllable modulation in the surface reactivity, but it also provides valuable information for the rational design of Pd-based catalysts for fuel cell applications.

  1. PDE Surface Generation with Combined Closed and Non-Closed Form Solutions

    Institute of Scientific and Technical Information of China (English)

    Jian-Jun Zhang; Li-Hua You

    2004-01-01

    Partial differential equations (PDEs) combined with suitably chosen boundary conditions are effective in creating free form surfaces. In this paper, a fourth order partial differential equation and boundary conditions up to tangential continuity are introduced. The general solution is divided into a closed form solution and a non-closed form one leading to a mixed solution to the PDE. The obtained solution is applied to a number of surface modelling examples including glass shape design, vase surface creation and arbitrary surface representation.

  2. Combined effect of electric field and surface modification on pool boiling of R-123

    OpenAIRE

    Ahmad, Syed Waqas

    2012-01-01

    This thesis was submitted for the degree of Doctor of Philosophy and awarded by Brunel University. The effect of surface modification and high intensity electric field (uniform and non – uniform) acting separately or in combination on pool boiling of R-123 is presented in this thesis. The effect of surface modification was investigated on saturated pool boiling of R-123 for five horizontal copper surfaces modified by different treatments, namely: an emery polished surface, a fine sandblast...

  3. Effects of Surface Composition on the Aerosolisation and Dissolution of Inhaled Antibiotic Combination Powders Consisting of Colistin and Rifampicin.

    Science.gov (United States)

    Wang, Wenbo; Zhou, Qi Tony; Sun, Si-Ping; Denman, John A; Gengenbach, Thomas R; Barraud, Nicolas; Rice, Scott A; Li, Jian; Yang, Mingshi; Chan, Hak-Kim

    2016-03-01

    Colistin is often the only effective antibiotic against the respiratory infections caused by multidrug-resistant Gram-negative bacteria. However, colistin-resistant multidrug-resistant isolates have been increasingly reported and combination therapy is preferred to combat resistance. In this study, five combination formulations containing colistin (COL) and rifampicin (RIF) were prepared by spray drying. The lowest minimum inhibitory concentration (MIC) value against Pseudomonas aeruginosa PAO1 was measured for the formulation of COL/RIF = 4:1 with relatively high emitted doses (over 80%) and satisfactory fine particle fractions (over 60%). Data from X-ray photoelectron spectroscopy (XPS) and nano-time-of-flight secondary ion mass spectrometry (ToF-SIMS) showed the surfaces of particles were mainly covered by rifampicin even for the formulation with a mass ratio of COL/RIF = 4:1. Because colistin is hygroscopic and rifampicin is hydrophobic, moisture absorption of combination formulations was significantly lower than the pure colistin formulation in the dynamic vapour sorption results. To investigate the dissolution characteristics, four dissolution test methods (diffusion Franz cell, modified Franz cell, flow-through and beaker methods) were employed and compared. The modified Franz cell method was selected to test the dissolution behaviour of aerosolised powder formulations to eliminate the effect of membrane on dissolution. The results showed that surface enrichment of hydrophobic rifampicin neither affected aerosolisation nor retarded dissolution rate of colistin in the combination formulations. For the first time, advanced surface characterisation techniques of XPS and ToF-SIMS have shown their capability to understand the effect of surface composition on the aerosolisation and dissolution of combination powders.

  4. Heat-transfer-method-based cell culture quality assay through cell detection by surface imprinted polymers.

    Science.gov (United States)

    Eersels, Kasper; van Grinsven, Bart; Khorshid, Mehran; Somers, Veerle; Püttmann, Christiane; Stein, Christoph; Barth, Stefan; Diliën, Hanne; Bos, Gerard M J; Germeraad, Wilfred T V; Cleij, Thomas J; Thoelen, Ronald; De Ceuninck, Ward; Wagner, Patrick

    2015-02-17

    Previous work has indicated that surface imprinted polymers (SIPs) allow for highly specific cell detection through macromolecular cell imprints. The combination of SIPs with a heat-transfer-based read-out technique has led to the development of a selective, label-free, low-cost, and user-friendly cell detection assay. In this study, the breast cancer cell line ZR-75-1 is used to assess the potential of the platform for monitoring the quality of a cell culture in time. For this purpose, we show that the proposed methodology is able to discriminate between the original cell line (adherent growth, ZR-75-1a) and a descendant cell line (suspension growth, ZR-75-1s). Moreover, ZR-75-1a cells were cultured for a prolonged period of time and analyzed using the heat-transfer method (HTM) at regular time intervals. The results of these experiments demonstrate that the thermal resistance (Rth) signal decays after a certain number of cell culture passages. This can likely be attributed to a compromised quality of the cell culture due to cross-contamination with the ZR-75-1s cell line, a finding that was confirmed by classical STR DNA profiling. The cells do not express the same functional groups on their membrane, resulting in a weaker bond between cell and imprint, enabling cell removal by mechanical friction, provided by flushing the measuring chamber with buffer solution. These findings were further confirmed by HTM and illustrate that the biomimetic sensor platform can be used as an assay for monitoring the quality of cell cultures in time.

  5. Basic surface properties of mononuclear cells from Didelphis marsupialis.

    Science.gov (United States)

    Nacife, V P; de Meirelles, M de N; Silva Filho, F C

    1998-01-01

    The electrostatic surface charge and surface tension of mononuclear cells/monocytes obtained from young and adult marsupials (Didelphis marsupialis) were investigated by using cationized ferritin and colloidal iron hydroxyde, whole cell electrophoresis, and measurements of contact angles. Anionic sites were found distributed throughout the entire investigated cell surfaces. The results revealed that the anionic character of the cells is given by electrostatic charges corresponding to -18.8 mV (cells from young animals) and -29.3 mV (cells from adult animals). The surface electrostatic charge decreased from 10 to 65.2% after treatment of the cells with each one of trypsin, neuraminidase and phospholipase C. The hydrophobic nature of the mononuclear cell surfaces studied by using the contact angle method revealed that both young and adult cells possess cell surfaces of high hidrofilicity since the angles formed with drops of saline water were 42.5 degrees and 40.8 degrees, respectively. Treatment of the cells with trypsin or neuraminidase rendered their surfaces more hydrophobic, suggesting that sialic acid-containing glycoproteins are responsible for most of the hydrophilicity observed in the mononuclear cell surfaces from D. marsupialis. PMID:9921307

  6. Basic Surface Properties of Mononuclear Cells from Didelphis marsupialis

    Directory of Open Access Journals (Sweden)

    Nacife Valéria Pereira

    1998-01-01

    Full Text Available The electrostatic surface charge and surface tension of mononuclear cells/monocytes obtained from young and adult marsupials (Didelphis marsupialis were investigated by using cationized ferritin and colloidal iron hydroxyde, whole cell electrophoresis, and measurements of contact angles. Anionic sites were found distributed throughout the entire investigated cell surfaces. The results revealed that the anionic character of the cells is given by electrostatic charges corresponding to -18.8 mV (cells from young animals and -29.3 mV (cells from adult animals. The surface electrostatic charge decreased from 10 to 65.2% after treatment of the cells with each one of trypsin, neuraminidase and phospholipase C. The hydrophobic nature of the mononuclear cell surfaces studied by using the contact angle method revealed that both young and adult cells possess cell surfaces of high hidrofilicity since the angles formed with drops of saline water were 42.5°and 40.8°, respectively. Treatment of the cells with trypsin or neuraminidase rendered their surfaces more hydrophobic, suggesting that sialic acid-containing glycoproteins are responsible for most of the hydrophilicity observed in the mononuclear cell surfaces from D. marsupialis.

  7. Surface cell differentiation controls tissue surface tension and tissue positioning during zebrafish gastrulation

    Science.gov (United States)

    Krens, S. F. G.

    2011-03-01

    Differences in tissue surface tension (TST) between different tissue types are thought to guide tissue organization and cell sorting in development. Measurements of TST have been useful to predict the outcome of in vitro cell sorting and envelopment experiments. However, the outcome of cell sorting experiments in vitro often substantially differs from tissue positioning in vivo, raising questions as to the actual contribution of TST to tissue positioning within the developing embryo. Here, we show that surface tension of germ layer tissues during zebrafish gastrulation critically relies on the differentiation of their surface cells. We also show that surface differentiation of the different germ layer tissues varies and is considerably different between the situation in vitro and in vivo, explaining the apparent dissimilar outcome of cell segregation between these two situations. To analyze germ layer TST as a function of surface cell differentiation, we interfere with surface cell properties of germ layer aggregates by misexpressing genes involved in surface cell differentiation specifically within surface cells using the GAL4-UAS system, and measure tissue surface tension using both parallel plate compression and micropipette aspiration techniques. Our data provides evidence in favor of a critical function of surface cell differentiation in modulating TST and subsequently tissue positioning within the developing embryo.

  8. Microfluidic single-cell transcriptional analysis rationally identifies novel surface marker profiles to enhance cell-based therapies.

    Science.gov (United States)

    Rennert, Robert C; Januszyk, Michael; Sorkin, Michael; Rodrigues, Melanie; Maan, Zeshaan N; Duscher, Dominik; Whittam, Alexander J; Kosaraju, Revanth; Chung, Michael T; Paik, Kevin; Li, Alexander Y; Findlay, Michael; Glotzbach, Jason P; Butte, Atul J; Gurtner, Geoffrey C

    2016-01-01

    Current progenitor cell therapies have only modest efficacy, which has limited their clinical adoption. This may be the result of a cellular heterogeneity that decreases the number of functional progenitors delivered to diseased tissue, and prevents correction of underlying pathologic cell population disruptions. Here, we develop a high-resolution method of identifying phenotypically distinct progenitor cell subpopulations via single-cell transcriptional analysis and advanced bioinformatics. When combined with high-throughput cell surface marker screening, this approach facilitates the rational selection of surface markers for prospective isolation of cell subpopulations with desired transcriptional profiles. We establish the usefulness of this platform in costly and highly morbid diabetic wounds by identifying a subpopulation of progenitor cells that is dysfunctional in the diabetic state, and normalizes diabetic wound healing rates following allogeneic application. We believe this work presents a logical framework for the development of targeted cell therapies that can be customized to any clinical application. PMID:27324848

  9. Nanofabrication of Nonfouling Surfaces for Micropatterning of Cell and Microtissue

    Directory of Open Access Journals (Sweden)

    Hidenori Otsuka

    2010-08-01

    Full Text Available Surface engineering techniques for cellular micropatterning are emerging as important tools to clarify the effects of the microenvironment on cellular behavior, as cells usually integrate and respond the microscale environment, such as chemical and mechanical properties of the surrounding fluid and extracellular matrix, soluble protein factors, small signal molecules, and contacts with neighboring cells. Furthermore, recent progress in cellular micropatterning has contributed to the development of cell-based biosensors for the functional characterization and detection of drugs, pathogens, toxicants, and odorants. In this regards, the ability to control shape and spreading of attached cells and cell-cell contacts through the form and dimension of the cell-adhesive patches with high precision is important. Commitment of stem cells to different specific lineages depends strongly on cell shape, implying that controlled microenvironments through engineered surfaces may not only be a valuable approach towards fundamental cell-biological studies, but also of great importance for the design of cell culture substrates for tissue engineering. To develop this kind of cellular microarray composed of a cell-resistant surface and cell attachment region, micropatterning a protein-repellent surface is important because cellular adhesion and proliferation are regulated by protein adsorption. The focus of this review is on the surface engineering aspects of biologically motivated micropatterning of two-dimensional surfaces with the aim to provide an introductory overview described in the literature. In particular, the importance of non-fouling surface chemistries is discussed.

  10. Combined immunotherapy and antiangiogenic therapy of cancer with microencapsulated cells.

    Science.gov (United States)

    Cirone, Pasquale; Bourgeois, Jacqueline M; Shen, Feng; Chang, Patricia L

    2004-10-01

    An alternative form of gene therapy involves immunoisolation of a nonautologous cell line engineered to secrete a therapeutic product. Encapsulation of these cells in a biocompatible polymer serves to protect these allogeneic cells from host-versus-graft rejection while recombinant products and nutrients are able to pass by diffusion. This strategy was applied to the treatment of cancer with some success by delivering either interleukin 2 or angiostatin. However, as cancer is a complex, multifactorial disease, a multipronged approach is now being developed to attack tumorigenesis via multiple pathways in order to improve treatment efficacy. A combination of immunotherapy with angiostatic therapy was investigated by treating B16-F0/neu melanoma-bearing mice with intraperitoneally implanted, microencapsulated mouse myoblasts (C2C12) genetically modified to deliver angiostatin and an interleukin 2 fusion protein (sFvIL-2). The combination treatment resulted in improved survival, delayed tumor growth, and increased histological indices of antitumor activity (apoptosis and necrosis). In addition to improved efficacy, the combination treatment also ameliorated some of the undesirable side effects from the individual treatments that have led to the previous failure of the single treatments, for example, inflammatory response to IL-2 or vascular mimicry due to angiostatin. In conclusion, the combination of immuno- and antiangiogenic therapies delivered by immunoisolated cells was superior to individual treatments for antitumorigenesis activity, not only because of their known mechanisms of action but also because of unexpected protection against the adverse side effects of the single treatments. Thus, the concept of a "cocktail" strategy, with microencapsulation delivering multiple antitumor recombinant molecules to improve efficacy, is validated. PMID:15585110

  11. Osteoblast cell response to surface-modified carbon nanotubes

    International Nuclear Information System (INIS)

    In order to investigate the interaction of cells with modified multi-walled carbon nanotubes (MWCNTs) for their potential biomedical applications, the MWCNTs were chemically modified with carboxylic acid groups (–COOH), polyvinyl alcohol (PVA) polymer and biomimetic apatite on their surfaces. Additionally, human osteoblast MG-63 cells were cultured in the presence of the surface-modified MWCNTs. The metabolic activities of osteoblastic cells, cell proliferation properties, as well as cell morphology were studied. The surface modification of MWCNTs with biomimetic apatite exhibited a significant increase in the cell viability of osteoblasts, up to 67.23%. In the proliferation phases, there were many more cells in the biomimetic apatite-modified MWCNT samples than in the MWCNTs–COOH. There were no obvious changes in cell morphology in osteoblastic MG-63 cells cultured in the presence of these chemically-modified MWCNTs. The surface modification of MWCNTs with apatite achieves an effective enhancement of their biocompatibility.

  12. INTEGRATED GASIFICATION COMBINED CYCLE PROJECT 2 MW FUEL CELL DEMONSTRATION

    Energy Technology Data Exchange (ETDEWEB)

    FuelCell Energy

    2005-05-16

    With about 50% of power generation in the United States derived from coal and projections indicating that coal will continue to be the primary fuel for power generation in the next two decades, the Department of Energy (DOE) Clean Coal Technology Demonstration Program (CCTDP) has been conducted since 1985 to develop innovative, environmentally friendly processes for the world energy market place. The 2 MW Fuel Cell Demonstration was part of the Kentucky Pioneer Energy (KPE) Integrated Gasification Combined Cycle (IGCC) project selected by DOE under Round Five of the Clean Coal Technology Demonstration Program. The participant in the CCTDP V Project was Kentucky Pioneer Energy for the IGCC plant. FuelCell Energy, Inc. (FCE), under subcontract to KPE, was responsible for the design, construction and operation of the 2 MW fuel cell power plant. Duke Fluor Daniel provided engineering design and procurement support for the balance-of-plant skids. Colt Engineering Corporation provided engineering design, fabrication and procurement of the syngas processing skids. Jacobs Applied Technology provided the fabrication of the fuel cell module vessels. Wabash River Energy Ltd (WREL) provided the test site. The 2 MW fuel cell power plant utilizes FuelCell Energy's Direct Fuel Cell (DFC) technology, which is based on the internally reforming carbonate fuel cell. This plant is capable of operating on coal-derived syngas as well as natural gas. Prior testing (1992) of a subscale 20 kW carbonate fuel cell stack at the Louisiana Gasification Technology Inc. (LGTI) site using the Dow/Destec gasification plant indicated that operation on coal derived gas provided normal performance and stable operation. Duke Fluor Daniel and FuelCell Energy developed a commercial plant design for the 2 MW fuel cell. The plant was designed to be modular, factory assembled and truck shippable to the site. Five balance-of-plant skids incorporating fuel processing, anode gas oxidation, heat recovery

  13. Dendritic Cell Responses to Surface Properties of Clinical Titanium Surfaces

    OpenAIRE

    Kou, Peng Meng; Schwartz, Zvi; Boyan, Barbara D; Babensee, Julia E.

    2010-01-01

    Dendritic cells (DCs) play pivotal roles in responding to foreign entities during an innate immune response and initiating effective adaptive immunity as well as maintaining immune tolerance. The sensitivity of DCs to foreign stimuli also makes them useful cells to assess the inflammatory response to biomaterials. Elucidating the material property-DC phenotype relationships using a well-defined biomaterial system is expected to provide criteria for immuno-modulatory biomaterial design. Clinic...

  14. Melittin interaction with sulfated cell surface sugars.

    Science.gov (United States)

    Klocek, Gabriela; Seelig, Joachim

    2008-03-01

    Melittin is a 26-residue cationic peptide with cytolytic and antimicrobial properties. Studies on the action mechanism of melittin have focused almost exclusively on the membrane-perturbing properties of this peptide, investigating in detail the melittin-lipid interaction. Here, we report physical-chemical studies on an alternative mechanism by which melittin could interact with the cell membrane. As the outer surface of many cells is decorated with anionic (sulfated) glycosaminoglycans (GAGs), a strong Coulombic interaction between the two oppositely charged molecules can be envisaged. Indeed, the present study using isothermal titration calorimetry reveals a high affinity of melittin for several GAGs, that is, heparan sulfate (HS), dermatan sulfate, and heparin. The microscopic binding constant of melittin for HS is 2.4 x 10 (5) M (-1), the reaction enthalpy is Delta H melittin (0) = -1.50 kcal/mol, and the peptide-to-HS stoichiometry is approximately 11 at 10 mM Tris, 100 mM NaCl at pH 7.4 and 28 degrees C. Delta H melittin (0) is characterized by a molar heat capacity of Delta C P (0) = -227 cal mol (-1) K (-1). The large negative heat capacity change indicates that hydrophobic interactions must also be involved in the binding of melittin to HS. Circular dichroism spectroscopy demonstrates that the binding of the peptide to HS induces a conformational change to a predominantly alpha-helical structure. A model for the melittin-HS complex is presented. Melittin binding was compared with that of magainin 2 and nisin Z to HS. Magainin 2 is known for its antimicrobial properties, but it does not cause lysis of the eukaryotic cells. Nisin Z shows activity against various Gram-positive bacteria. Isothermal titration calorimetry demonstrates that magainin 2 and nisin Z do not bind to HS (5-50 degrees C, 10 mM Tris, and 100 mM NaCl at pH 7.4). PMID:18220363

  15. Calreticulin: Roles in Cell-Surface Protein Expression

    Directory of Open Access Journals (Sweden)

    Yue Jiang

    2014-09-01

    Full Text Available In order to perform their designated functions, proteins require precise subcellular localizations. For cell-surface proteins, such as receptors and channels, they are able to transduce signals only when properly targeted to the cell membrane. Calreticulin is a multi-functional chaperone protein involved in protein folding, maturation, and trafficking. However, evidence has been accumulating that calreticulin can also negatively regulate the surface expression of certain receptors and channels. In these instances, depletion of calreticulin enhances cell-surface expression and function. In this review, we discuss the role of calreticulin with a focus on its negative effects on the expression of cell-surface proteins.

  16. A combined gas cooled nuclear reactor and fuel cell cycle

    Science.gov (United States)

    Palmer, David J.

    Rising oil costs, global warming, national security concerns, economic concerns and escalating energy demands are forcing the engineering communities to explore methods to address these concerns. It is the intention of this thesis to offer a proposal for a novel design of a combined cycle, an advanced nuclear helium reactor/solid oxide fuel cell (SOFC) plant that will help to mitigate some of the above concerns. Moreover, the adoption of this proposal may help to reinvigorate the Nuclear Power industry while providing a practical method to foster the development of a hydrogen economy. Specifically, this thesis concentrates on the importance of the U.S. Nuclear Navy adopting this novel design for its nuclear electric vessels of the future with discussion on efficiency and thermodynamic performance characteristics related to the combined cycle. Thus, the goals and objectives are to develop an innovative combined cycle that provides a solution to the stated concerns and show that it provides superior performance. In order to show performance, it is necessary to develop a rigorous thermodynamic model and computer program to analyze the SOFC in relation with the overall cycle. A large increase in efficiency over the conventional pressurized water reactor cycle is realized. Both sides of the cycle achieve higher efficiencies at partial loads which is extremely important as most naval vessels operate at partial loads as well as the fact that traditional gas turbines operating alone have poor performance at reduced speeds. Furthermore, each side of the cycle provides important benefits to the other side. The high temperature exhaust from the overall exothermic reaction of the fuel cell provides heat for the reheater allowing for an overall increase in power on the nuclear side of the cycle. Likewise, the high temperature helium exiting the nuclear reactor provides a controllable method to stabilize the fuel cell at an optimal temperature band even during transients helping

  17. Combined toxicity of heavy metal mixtures in liver cells.

    Science.gov (United States)

    Lin, Xialu; Gu, Yuanliang; Zhou, Qi; Mao, Guochuan; Zou, Baobo; Zhao, Jinshun

    2016-09-01

    With rapid industrialization, China is now facing great challenges in heavy metal contamination in the environment. Human exposure to heavy metals through air, water and food commonly involves a mixture consisting of multiple heavy metals. In this study, eight common heavy metals (Pb, Cd, Hg, Cu, Zn, Mn, Cr, Ni) that cause environmental contamination were selected to investigate the combined toxicity of different heavy metal mixtures in HL7702 cells. Toxicity (24 h LC50 ) of each individual metal on the cells ranked Hg > Cr = Cd > Cu > Zn > Ni > Mn > Pb; toxicity of the different mixtures ranked: M5 > M3PbHgCd > M5+Mn > M5+Cu > M2CdNi > M4A > M8-Mn > M8 > M5+Zn > M4B > M8-Cr > M8-Zn > M8-Cu > M8-Pb > M8-Cd > M8-Hg > M8-Ni > M3PbHgNi > M3CuZnMn. The cytotoxicity data of individual metals were successfully used to build the additive models of two- to eight-component metal mixtures. The comparison between additive model and combination model or partly additive model was useful to evaluate the combined effects in mixture. Synergistic, antagonistic or additive effects of the toxicity were observed in different mixtures. These results suggest that the combined effects should be considered in the risk assessment of heavy metal co-exposure, and more comprehensive investigations on the combined effects of different heavy metal mixtures are needed in the future. Copyright © 2016 John Wiley & Sons, Ltd.

  18. Response of breast cancer cells and cancer stem cells to metformin and hyperthermia alone or combined.

    Directory of Open Access Journals (Sweden)

    Hyemi Lee

    Full Text Available Metformin, the most widely prescribed drug for treatment of type 2 diabetes, has been shown to exert significant anticancer effects. Hyperthermia has been known to kill cancer cells and enhance the efficacy of various anti-cancer drugs and radiotherapy. We investigated the combined effects of metformin and hyperthermia against MCF-7 and MDA-MB-231 human breast cancer cell, and MIA PaCa-2 human pancreatic cancer cells. Incubation of breast cancer cells with 0.5-10 mM metformin for 48 h caused significant clonogenic cell death. Culturing breast cancer cells with 30 µM metformin, clinically relevant plasma concentration of metformin, significantly reduced the survival of cancer cells. Importantly, metformin was preferentially cytotoxic to CD44(high/CD24(low cells of MCF-7 cells and, CD44(high/CD24(high cells of MIA PaCa-2 cells, which are known to be cancer stem cells (CSCs of MCF-7 cells and MIA PaCa-2 cells, respectively. Heating at 42°C for 1 h was slightly toxic to both cancer cells and CSCs, and it markedly enhanced the efficacy of metformin to kill cancer cells and CSCs. Metformin has been reported to activate AMPK, thereby suppressing mTOR, which plays an important role for protein synthesis, cell cycle progression, and cell survival. For the first time, we show that hyperthermia activates AMPK and inactivates mTOR and its downstream effector S6K. Furthermore, hyperthermia potentiated the effect of metformin to activate AMPK and inactivate mTOR and S6K. Cell proliferation was markedly suppressed by metformin or combination of metformin and hyperthermia, which could be attributed to activation of AMPK leading to inactivation of mTOR. It is conclude that the effects of metformin against cancer cells including CSCs can be markedly enhanced by hyperthermia.

  19. Targeting Negative Surface Charges of Cancer Cells by Multifunctional Nanoprobes

    Science.gov (United States)

    Chen, Bingdi; Le, Wenjun; Wang, Yilong; Li, Zhuoquan; Wang, Dong; Ren, Lei; Lin, Ling; Cui, Shaobin; Hu, Jennifer J.; Hu, Yihui; Yang, Pengyuan; Ewing, Rodney C.; Shi, Donglu; Cui, Zheng

    2016-01-01

    A set of electrostatically charged, fluorescent, and superparamagnetic nanoprobes was developed for targeting cancer cells without using any molecular biomarkers. The surface electrostatic properties of the established cancer cell lines and primary normal cells were characterized by using these nanoprobes with various electrostatic signs and amplitudes. All twenty two randomly selected cancer cell lines of different organs, but not normal control cells, bound specifically to the positively charged nanoprobes. The relative surface charges of cancer cells could be quantified by the percentage of cells captured magnetically. The activities of glucose metabolism had a profound impact on the surface charge level of cancer cells. The data indicate that an elevated glycolysis in the cancer cells led to a higher level secretion of lactate. The secreted lactate anions are known to remove the positive ions, leaving behind the negative changes on the cell surfaces. This unique metabolic behavior is responsible for generating negative cancer surface charges in a perpetuating fashion. The metabolically active cancer cells are shown to a unique surface electrostatic pattern that can be used for recovering cancer cells from the circulating blood and other solutions. PMID:27570558

  20. Targeting Negative Surface Charges of Cancer Cells by Multifunctional Nanoprobes.

    Science.gov (United States)

    Chen, Bingdi; Le, Wenjun; Wang, Yilong; Li, Zhuoquan; Wang, Dong; Ren, Lei; Lin, Ling; Cui, Shaobin; Hu, Jennifer J; Hu, Yihui; Yang, Pengyuan; Ewing, Rodney C; Shi, Donglu; Cui, Zheng

    2016-01-01

    A set of electrostatically charged, fluorescent, and superparamagnetic nanoprobes was developed for targeting cancer cells without using any molecular biomarkers. The surface electrostatic properties of the established cancer cell lines and primary normal cells were characterized by using these nanoprobes with various electrostatic signs and amplitudes. All twenty two randomly selected cancer cell lines of different organs, but not normal control cells, bound specifically to the positively charged nanoprobes. The relative surface charges of cancer cells could be quantified by the percentage of cells captured magnetically. The activities of glucose metabolism had a profound impact on the surface charge level of cancer cells. The data indicate that an elevated glycolysis in the cancer cells led to a higher level secretion of lactate. The secreted lactate anions are known to remove the positive ions, leaving behind the negative changes on the cell surfaces. This unique metabolic behavior is responsible for generating negative cancer surface charges in a perpetuating fashion. The metabolically active cancer cells are shown to a unique surface electrostatic pattern that can be used for recovering cancer cells from the circulating blood and other solutions. PMID:27570558

  1. How cells tiptoe on adhesive surfaces before sticking

    CERN Document Server

    Pierres, Anne; Touchard, Dominique; Bongrand, Pierre

    2008-01-01

    Cell membranes are studded with protrusions that were thoroughly analyzed with electron microscopy. However, the nanometer-scale three-dimensional motions generated by cell membranes to fit the topography of foreign surfaces and initiate adhesion remain poorly understood. Here, we describe the dynamics of surface deformations displayed by monocytic cells bumping against fibronectin-coated surfaces. We observed membrane undulations with typically 5 nm amplitude and 5-10 second lifetime. Cell membranes behaved as independent units of micrometer size. Cells detected the presence of foreign surfaces at 50 nm separation, resulting in time-dependent amplification of membrane undulations. Molecular contact then ensued with apparent cell-membrane separation of 30-40 nm, and this distance steadily decreased during the following tens of seconds. Contact maturation was associated with in-plane egress of bulky molecules and robust membrane fluctuations. Thus, membrane undulations may be the major determinant of cell sens...

  2. Northern South China Sea Surface Circulation and its Variability Derived by Combining Satellite Altimetry and Surface Drifter Data

    Directory of Open Access Journals (Sweden)

    N. Peter Benny

    2015-01-01

    Full Text Available The present study analyses the mean and seasonal mesoscale surface circulation of the Northern South China Sea (NSCS and determines the influence of El Niño/SouthernNiño/Southern Oscillation (ENSO. High resolution Eulerian velocity field is derived by combining the available satellite tracked surface drifter data with satellite altimetry during 1993 - 2012. The wind driven current is computed employing the weekly ocean surface mean wind fields derived from the scatterometers on board ERS 1/2, QuikSCAT and ASCAT. The derived mean velocity field exhibits strong boundary currents and broad zonal flow across NSCS. The anomalous field is quite strong in the southern part and the Seasonal circulation clearly depicts the monsoonal forcing. Eddy Kinetic Energy (EKE distribution and its spatial and temporal structures are determined employing Empirical Orthogonal Function (EOF analysis. The ENSO influence on NSCS surface circulation has been analyzed using monthly absolute geostrophic velocity fields during 1996 - 1999.

  3. Molecularly engineered surfaces for cell biology: from static to dynamic surfaces.

    Science.gov (United States)

    Gooding, J Justin; Parker, Stephen G; Lu, Yong; Gaus, Katharina

    2014-04-01

    Surfaces with a well-defined presentation of ligands for receptors on the cell membrane can serve as models of the extracellular matrix for studying cell adhesion or as model cell surfaces for exploring cell-cell contacts. Because such surfaces can provide exquisite control over, for example, the density of these ligands or when the ligands are presented to the cell, they provide a very precise strategy for understanding the mechanisms by which cells respond to external adhesive cues. In the present feature article, we present an overview of the basic biology of cell adhesion before discussing surfaces that have a static presentation of immobile ligands. We outline the biological information that such surfaces have given us, before progressing to recently developed switchable surfaces and surfaces that mimic the lipid bilayer, having adhesive ligands that can move around the membrane and be remodeled by the cell. Finally, the feature article closes with some of the biological information that these new types of surfaces could provide.

  4. Combining oxygen plasma treatment with anchorage of cationized gelatin for enhancing cell affinity of poly(lactide-co-glycolide).

    Science.gov (United States)

    Shen, Hong; Hu, Xixue; Yang, Fei; Bei, Jianzhong; Wang, Shenguo

    2007-10-01

    Surface characteristics greatly influence attachment and growth of cells on biomaterials. Although polylactone-type biodegradable polymers have been widely used as scaffold materials for tissue engineering, lack of cell recognition sites, poor hydrophilicity and low surface energy lead to a bad cell affinity of the polymers, which limit the usage of polymers as scaffolds in tissue engineering. In the present study, surface of poly (L-lactide-co-glycolide) (PLGA) was modified by a method of combining oxygen plasma treatment with anchorage of cationized gelatin. Modification effect of the method was compared with other methods of oxygen plasma treatment, cationized gelatin or gelatin coating and combining oxygen plasma treatment with anchorage of gelatin. The change of surface property was compared by contact angles, surface energy, X-ray photoelectron spectra (XPS), attenuated total reflection-Fourier transform infrared spectroscopy (ATR-FTIR) and scanning electron microscopy (SEM) measurement. The optimum oxygen pretreatment time determined by surface energy was 10 min when the power was 50 W and the oxygen pressure was 20 Pa. Analysis of the stability of gelatin and cationized gelatin anchored on PLGA by XPS, ATR-FTIR, contact angles and surface energy measurement indicated the cationized gelatin was more stable than gelatin. The result using mouse NIH 3T3 fibroblasts as model cells to evaluate cell affinity in vitro showed the cationized gelatin-anchored PLGA (OCG-PLGA) was more favorable for cell attachment and growth than oxygen plasma treated PLGA (O-PLGA) and gelatin-anchored PLGA (OG-PLGA). Moreover cell affinity of OCG-PLGA could match that of collagen-anchored PLGA (AC-PLGA). So the surface modification method combining oxygen plasma treatment with anchorage of cationized gelatin provides a universally effective way to enhance cell affinity of polylactone-type biodegradable polymers. PMID:17618682

  5. Combined study of evaporation from liquid surface by background oriented schlieren, infrared thermal imaging and numerical simulation

    Directory of Open Access Journals (Sweden)

    Plaksina Yu.Yu.

    2013-04-01

    Full Text Available Temperature fields in evaporating liquids are measured by simultaneous use of Background Oriented Schlieren (BOS technique for the side view and IR thermal imaging for the surface distribution. Good agreement between the two methods is obtained with typical measurement error less than 0.1 K. Two configurations of surface layer are observed: thermocapillary convection state with moving liquid surface and small thermal cells, associated with Marangoni convection, and “cool skin” with negligible velocity at the surface, larger cells and dramatic increase of velocity within 0.1 mm layer beneath the surface. These configurations are shown to be formed in various liquids (water with various degrees of purification, ethanol, butanol, decane, kerosene, glycerine depending rather on initial conditions and ambient parameters than on the liquid. Water, which has been considered as the liquid without observable Marangoni convection, actually can exhibit both kinds of behavior during the same experimental run. Evaporation is also studied by means of numerical simulations. Separate problemsin air and liquid are considered, with thermal imaging data of surface temperature making the separation possible. It is shown that evaporation rate can be predicted by numerical simulation of the air side with appropriate boundary conditions. Comparison is made with known empirical correlations for Sherwood-Rayleigh relationship. Numerical simulations of water-side problem reveal the issue of velocity boundary conditions at the free surface, determining the structure of surface layer. Flow field similar to observed in the experiments is obtained with special boundary conditions of third kind, presenting a combination of no-slip and surface tension boundary conditions.

  6. Microplicae: specialized surface structure of epithelial cells of wet-surfaced oral mucosa

    NARCIS (Netherlands)

    P. Asikainen; E. Sirviö; J.J.W. Mikkonen; S.P. Singh; E.A.J.M. Schulten; C.M. ten Bruggenkate; A.P. Koistinen; A.M. Kullaa

    2015-01-01

    The surface structure of the superficial cells of the oral mucosa is decorated with numerous membrane ridges, termed microplicae (MPLs). The MPL structure is typical of the epithelial surfaces that are covered with protective mucus. Cell membrane MPLs are no longer seen as passive consequences of ce

  7. FABRICATION AND BIOCOMPATIBILITY OF CELL OUTER MEMBRANE MIMETIC SURFACES

    Institute of Scientific and Technical Information of China (English)

    Ming-ming Zong; Yong-kuan Gong

    2011-01-01

    The surface design used for improving biocompatibility is one of the most important issues for the fabrication of medical devices. For mimicking the ideal surface structure of cell outer membrane, a large number of polymers bearing phosphorylcholine (PC) groups have been employed to modify the surfaces of biomaterials and medical devices. It has been demonstrated that the biocompatibility of the modified materials whose surface is required to interact with a living organism has been obviously improved by introducing PC groups. In this review, the fabrication strategies of cell outer membrane mimetic surfaces and their resulted biocompatibilities were summarized.

  8. Combined single cell AFM manipulation and TIRFM for probing the molecular stability of multilayer fibrinogen matrices

    Energy Technology Data Exchange (ETDEWEB)

    Christenson, W. [Department of Physics, Arizona State University, Tempe, AZ 85287 (United States); Center for Biological Physics, Arizona State University, Tempe, AZ 85287 (United States); Yermolenko, I. [Department of Physics, Arizona State University, Tempe, AZ 85287 (United States); School of Life Sciences, Arizona State University, Tempe, AZ 85287 (United States); Plochberger, B. [Department of Physics, Arizona State University, Tempe, AZ 85287 (United States); Camacho-Alanis, F.; Ros, A. [Department of Chemistry and Biochemistry, Arizona State University, Tempe, AZ 85287 (United States); Ugarova, T.P. [School of Life Sciences, Arizona State University, Tempe, AZ 85287 (United States); Ros, R., E-mail: Robert.Ros@asu.edu [Department of Physics, Arizona State University, Tempe, AZ 85287 (United States); Center for Biological Physics, Arizona State University, Tempe, AZ 85287 (United States)

    2014-01-15

    Adsorption of fibrinogen on various surfaces produces a nanoscale multilayer matrix, which strongly reduces the adhesion of platelets and leukocytes with implications for hemostasis and blood compatibility of biomaterials. The nonadhesive properties of fibrinogen matrices are based on their extensibility, ensuing the inability to transduce strong mechanical forces via cellular integrins and resulting in weak intracellular signaling. In addition, reduced cell adhesion may arise from the weaker associations between fibrinogen molecules in the superficial layers of the matrix. Such reduced stability would allow integrins to pull fibrinogen molecules out of the matrix with comparable or smaller forces than required to break integrin–fibrinogen bonds. To examine this possibility, we developed a method based on the combination of total internal reflection fluorescence microscopy, single cell manipulation with an atomic force microscope and microcontact printing to study the transfer of fibrinogen molecules out of a matrix onto cells. We calculated the average fluorescence intensities per pixel for wild-type HEK 293 (HEK WT) and HEK 293 cells expressing leukocyte integrin Mac-1 (HEK Mac-1) before and after contact with multilayered matrices of fluorescently labeled fibrinogen. For contact times of 500 s, HEK Mac-1 cells show a median increase of 57% of the fluorescence intensity compared to 6% for HEK WT cells. The results suggest that the integrin Mac-1-fibrinogen interactions are stronger than the intermolecular fibrinogen interactions in the superficial layer of the matrix. The low mechanical stability of the multilayer fibrinogen surface may contribute to the reduced cell adhesive properties of fibrinogen-coated substrates. We anticipate that the described method can be applied to various cell types to examine their integrin-mediated adhesion to the extracellular matrices with a variable protein composition. - Highlights: • We present a method combining

  9. Novel eukaryotic enzymes modifying cell-surface biopolymers

    Directory of Open Access Journals (Sweden)

    Aravind L

    2010-01-01

    Full Text Available Abstract Background Eukaryotic extracellular matrices such as proteoglycans, sclerotinized structures, mucus, external tests, capsules, cell walls and waxes contain highly modified proteins, glycans and other composite biopolymers. Using comparative genomics and sequence profile analysis we identify several novel enzymes that could be potentially involved in the modification of cell-surface glycans or glycoproteins. Results Using sequence analysis and conservation we define the acyltransferase domain prototyped by the fungal Cas1p proteins, identify its active site residues and unify them to the superfamily of classical 10TM acyltransferases (e.g. oatA. We also identify a novel family of esterases (prototyped by the previously uncharacterized N-terminal domain of Cas1p that have a similar fold as the SGNH/GDSL esterases but differ from them in their conservation pattern. Conclusions We posit that the combined action of the acyltransferase and esterase domain plays an important role in controlling the acylation levels of glycans and thereby regulates their physico-chemical properties such as hygroscopicity, resistance to enzymatic hydrolysis and physical strength. We present evidence that the action of these novel enzymes on glycans might play an important role in host-pathogen interaction of plants, fungi and metazoans. We present evidence that in plants (e.g. PMR5 and ESK1 the regulation of carbohydrate acylation by these acylesterases might also play an important role in regulation of transpiration and stress resistance. We also identify a subfamily of these esterases in metazoans (e.g. C7orf58, which are fused to an ATP-grasp amino acid ligase domain that is predicted to catalyze, in certain animals, modification of cell surface polymers by amino acid or peptides. Reviewers This article was reviewed by Gaspar Jekely and Frank Eisenhaber

  10. Novel eukaryotic enzymes modifying cell-surface biopolymers

    Science.gov (United States)

    2010-01-01

    Background Eukaryotic extracellular matrices such as proteoglycans, sclerotinized structures, mucus, external tests, capsules, cell walls and waxes contain highly modified proteins, glycans and other composite biopolymers. Using comparative genomics and sequence profile analysis we identify several novel enzymes that could be potentially involved in the modification of cell-surface glycans or glycoproteins. Results Using sequence analysis and conservation we define the acyltransferase domain prototyped by the fungal Cas1p proteins, identify its active site residues and unify them to the superfamily of classical 10TM acyltransferases (e.g. oatA). We also identify a novel family of esterases (prototyped by the previously uncharacterized N-terminal domain of Cas1p) that have a similar fold as the SGNH/GDSL esterases but differ from them in their conservation pattern. Conclusions We posit that the combined action of the acyltransferase and esterase domain plays an important role in controlling the acylation levels of glycans and thereby regulates their physico-chemical properties such as hygroscopicity, resistance to enzymatic hydrolysis and physical strength. We present evidence that the action of these novel enzymes on glycans might play an important role in host-pathogen interaction of plants, fungi and metazoans. We present evidence that in plants (e.g. PMR5 and ESK1) the regulation of carbohydrate acylation by these acylesterases might also play an important role in regulation of transpiration and stress resistance. We also identify a subfamily of these esterases in metazoans (e.g. C7orf58), which are fused to an ATP-grasp amino acid ligase domain that is predicted to catalyze, in certain animals, modification of cell surface polymers by amino acid or peptides. Reviewers This article was reviewed by Gaspar Jekely and Frank Eisenhaber PMID:20056006

  11. The Use of Yeast Surface Display in Biofuel Cells.

    Science.gov (United States)

    Szczupak, Alon; Alfonta, Lital

    2015-01-01

    Biofuel cells are electrochemical devices which convert chemical energy to electricity using biochemical pathways and redox enzymes. In enzymatic fuel cells purified redox enzymes catalyze the reactions in the anode and cathode compartments whereas in microbial fuel cells (MFCs) the entire metabolism of the microorganisms is exploited. Here, a hybrid biofuel cell concept is presented, which is based on yeast surface display (YSD) of redox enzymes to catalyze the different cell reactions. PMID:26060081

  12. Artemether Combined with shRNA Interference of Vascular Cell Adhesion Molecule-1 Significantly Inhibited the Malignant Biological Behavior of Human Glioma Cells

    OpenAIRE

    Ying-Bin Wang; Yi Hu; Zhen Li; Ping Wang; Yi-Xue Xue; Yi-Long Yao; Bo Yu; Yun-Hui Liu

    2013-01-01

    Artemether is the derivative extracted from Chinese traditional herb and originally used for malaria. Artemether also has potential therapeutic effects against tumors. Vascular cell adhesion molecule-1 (VCAM-1) is an important cell surface adhesion molecule associated with malignancy of gliomas. In this work, we investigated the role and mechanism of artemether combined with shRNA interference of VCAM-1 (shRNA-VCAM-1) on the migration, invasion and apoptosis of glioma cells. U87 human glioma ...

  13. Influence of Combined Hard and Fine Machining on the Surface Properties of Cemented Carbides

    Directory of Open Access Journals (Sweden)

    U. Engel

    2012-09-01

    Full Text Available As a result of recent developments in cold forging cemented carbides are increasingly used as tool materials. Due to their high hardness only electrical discharge machining (EDM and grinding are suitable for tool machining. The structure of tool surface has significant influence on dominating failure mechanisms wear and fatigue. For improvement of tribological conditions the surface is polished in a finale processing step. The result of hard and fine machining is a specific combination of coarse and fine structure which is determined by processing parameters. The different surface structures lead to a particular tool behavior in forming process. This paper aims to show the influence of combined hard and fine machining on the surface properties of cemented carbides.

  14. Smooth Muscle Cell Functionality on Collagen Immobilized Polycaprolactone Nanowire Surfaces

    Directory of Open Access Journals (Sweden)

    Victoria Leszczak

    2014-05-01

    Full Text Available Inhibition of smooth muscle cell (SMC proliferation and preservation of a differentiated state are important aspects in the management, avoidance and progression of vascular diseases. An understanding of the interaction between SMCs and the biomaterial involved is essential for a successful implant. In this study, we have developed collagen immobilized nanostructured surfaces with controlled arrays of high aspect ratio nanowires for the growth and maintenance of human aortic SMCs. The nanowire surfaces were fabricated from polycaprolactone and were immobilized with collagen. The objective of this study is to reveal how SMCs interact with collagen immobilized nanostructures. The results indicate significantly higher cellular adhesion on nanostructured and collagen immobilized surfaces; however, SMCs on nanostructured surfaces exhibit a more elongated phenotype. The reduction of MTT was significantly lower on nanowire (NW and collagen immobilized NW (colNW surfaces, suggesting that SMCs on nanostructured surfaces may be differentiated and slowly dividing. Scanning electron microscopy results reveal that SMCs on nanostructured surfaces are more elongated and that cells are interacting with the nano-features on the surface. After providing differentiation cues, heavy chain myosin and calponin, specific to a contractile SMC phenotype, are upregulated on collagen immobilized surfaces. These results suggest that nanotopography affects cell adhesion, proliferation, as well as cell elongation, while collagen immobilized surfaces greatly affect cell differentiation.

  15. Cell manipulation tool with combined microwell array and optical tweezers for cell isolation and deposition

    International Nuclear Information System (INIS)

    Isolation from rare cells and deposition of sorted cells with high accuracy for further study are critical to a wide range of biomedical applications. In the current paper, we report an automated cell manipulation tool with combined optical tweezers and a uniquely designed microwell array, which functions for recognition, isolation, assembly, transportation and deposition of the interesting cells. The microwell array allows the passive hydrodynamic docking of cells, while offering the opportunity to inspect the interesting cell phenotypes with high spatio-temporal resolution based on the flexible image processing technique. In addition, dynamic and parallel cell manipulation in three dimensions can realize the target cell levitation from microwell and pattern assembly with multiple optical traps. Integrated with the programmed motorized stage, the optically levitated and assembled cells can be transported and deposited to the predefined microenvironment, so the tool can facilitate the integration of other on-chip functionalities for further study without removing these isolated cells from the chip. Experiments on human embryonic stem cells and yeast cells are performed to demonstrate the effectiveness of the proposed cell manipulation tool. Besides the application to cell isolation and deposition, three other biological applications with this tool are also presented. (paper)

  16. Cell Surface Proteome of Dental Pulp Stem Cells Identified by Label-Free Mass Spectrometry

    Science.gov (United States)

    Niehage, Christian; Karbanová, Jana; Steenblock, Charlotte

    2016-01-01

    Multipotent mesenchymal stromal cells (MSCs) are promising tools for regenerative medicine. They can be isolated from different sources based on their plastic-adherence property. The identification of reliable cell surface markers thus becomes the Holy Grail for their prospective isolation. Here, we determine the cell surface proteomes of human dental pulp-derived MSCs isolated from single donors after culture expansion in low (2%) or high (10%) serum-containing media. Cell surface proteins were tagged on intact cells using cell impermeable, cleavable sulfo-NHS-SS-biotin, which allows their enrichment by streptavidin pull-down. For the proteomic analyses, we first compared label-free methods to analyze cell surface proteomes i.e. composition, enrichment and proteomic differences, and we developed a new mathematical model to determine cell surface protein enrichment using a combinatorial gene ontology query. Using this workflow, we identified 101 cluster of differentiation (CD) markers and 286 non-CD cell surface proteins. Based on this proteome profiling, we identified 14 cell surface proteins, which varied consistently in abundance when cells were cultured under low or high serum conditions. Collectively, our analytical methods provide a basis for identifying the cell surface proteome of dental pulp stem cells isolated from single donors and its evolution during culture or differentiation. Our data provide a comprehensive cell surface proteome for the precise identification of dental pulp-derived MSC populations and their isolation for potential therapeutic intervention. PMID:27490675

  17. Cell Surface Proteome of Dental Pulp Stem Cells Identified by Label-Free Mass Spectrometry.

    Science.gov (United States)

    Niehage, Christian; Karbanová, Jana; Steenblock, Charlotte; Corbeil, Denis; Hoflack, Bernard

    2016-01-01

    Multipotent mesenchymal stromal cells (MSCs) are promising tools for regenerative medicine. They can be isolated from different sources based on their plastic-adherence property. The identification of reliable cell surface markers thus becomes the Holy Grail for their prospective isolation. Here, we determine the cell surface proteomes of human dental pulp-derived MSCs isolated from single donors after culture expansion in low (2%) or high (10%) serum-containing media. Cell surface proteins were tagged on intact cells using cell impermeable, cleavable sulfo-NHS-SS-biotin, which allows their enrichment by streptavidin pull-down. For the proteomic analyses, we first compared label-free methods to analyze cell surface proteomes i.e. composition, enrichment and proteomic differences, and we developed a new mathematical model to determine cell surface protein enrichment using a combinatorial gene ontology query. Using this workflow, we identified 101 cluster of differentiation (CD) markers and 286 non-CD cell surface proteins. Based on this proteome profiling, we identified 14 cell surface proteins, which varied consistently in abundance when cells were cultured under low or high serum conditions. Collectively, our analytical methods provide a basis for identifying the cell surface proteome of dental pulp stem cells isolated from single donors and its evolution during culture or differentiation. Our data provide a comprehensive cell surface proteome for the precise identification of dental pulp-derived MSC populations and their isolation for potential therapeutic intervention. PMID:27490675

  18. Cell multiplication following partial enzymatic removal of surface coat.

    Science.gov (United States)

    Wyroba, E

    1978-08-01

    Treatment of Paramecium aurelia with trypsin or pronase (1 mg per 10(5) cells, at 0 to 4 degrees C) partially removes the surface coat and modifies significantly multiplication of cells. The division rate after 24 hours of cultivation is diminished approximately twice in the case of pronase-treated cells and 1.5 for tyrpsin-digested ciliates as compared with the control. On the second day the division rate increases rapidly and number of cell divisions exceeds the values observed in the control. After 72 hours of cultivation the division rate in both untreated and enzyme-treated cells is almost the same. It is concluded that the observed inhibition of cell fission results from the enzymatic removal of the surface coat--the integrity of this surface coat seems to be necessary in the process of cell division. The influence of environmental factors on the rate of growth is presented.

  19. Surface-modified gold nanorods for specific cell targeting

    Science.gov (United States)

    Wang, Chan-Ung; Arai, Yoshie; Kim, Insun; Jang, Wonhee; Lee, Seonghyun; Hafner, Jason H.; Jeoung, Eunhee; Jung, Deokho; Kwon, Youngeun

    2012-05-01

    Gold nanoparticles (GNPs) have unique properties that make them highly attractive materials for developing functional reagents for various biomedical applications including photothermal therapy, targeted drug delivery, and molecular imaging. For in vivo applications, GNPs need to be prepared with very little or negligible cytotoxicitiy. Most GNPs are, however, prepared using growth-directing surfactants such as cetyl trimethylammonium bromide (CTAB), which are known to have considerable cytotoxicity. In this paper, we describe an approach to remove CTAB to a non-toxic concentration. We optimized the conditions for surface modification with methoxypolyethylene glycol thiol (mPEG), which replaced CTAB and formed a protective layer on the surface of gold nanorods (GNRs). The cytotoxicities of pristine and surface-modified GNRs were measured in primary human umbilical vein endothelial cells and human cell lines derived from hepatic carcinoma cells, embryonic kidney cells, and thyroid papillary carcinoma cells. Cytotoxicity assays revealed that treating cells with GNRs did not significantly affect cell viability except for thyroid papillary carcinoma cells. Thyroid cancer cells were more susceptible to residual CTAB, so CTAB had to be further removed by dialysis in order to use GNRs for thyroid cell targeting. PEGylated GNRs are further modified to present monoclonal antibodies that recognize a specific surface marker, Na-I symporter, for thyroid cells. Antibody-conjugated GNRs specifically targeted human thyroid cells in vitro.

  20. Solitary wave propagation in surface stabilized ferroelectric liquid crystal cells

    OpenAIRE

    VIJ, JAGDISH; Song, Jang-Kun

    2008-01-01

    PUBLISHED Solitary wave propagation in surface stabilized ferroelectric liquid crystal cells controlled by surface anchoring of the alignment layers is investigated for different conditions of alignment on the two opposite surfaces. We show that the critical field Ec, where the speed of the solitary wave becomes zero, is finite for asymmetric alignment on two surfaces. We also show that the polar anchoring energy difference (Deltawp) between the alignment layers can be calculated by measur...

  1. Fluid dynamics and noise in bacterial cell-cell and cell-surface scattering

    CERN Document Server

    Drescher, Knut; Cisneros, Luis H; Ganguly, Sujoy; Goldstein, Raymond E; 10.1073/pnas.1019079108

    2011-01-01

    Bacterial processes ranging from gene expression to motility and biofilm formation are constantly challenged by internal and external noise. While the importance of stochastic fluctuations has been appreciated for chemotaxis, it is currently believed that deterministic long-range fluid dynamical effects govern cell-cell and cell-surface scattering - the elementary events that lead to swarming and collective swimming in active suspensions and to the formation of biofilms. Here, we report the first direct measurements of the bacterial flow field generated by individual swimming Escherichia coli both far from and near to a solid surface. These experiments allowed us to examine the relative importance of fluid dynamics and rotational diffusion for bacteria. For cell-cell interactions it is shown that thermal and intrinsic stochasticity drown the effects of long-range fluid dynamics, implying that physical interactions between bacteria are determined by steric collisions and near-field lubrication forces. This dom...

  2. Interaction of Epithelial Cells with Surfaces and Surfaces Decorated by Molecules

    CERN Document Server

    Martini, Daniele; Beil, Michael; Paust, T; Huang, C; Moosmann, M; Jin, J; Heiler, T; Gröger, R; Schimmel, Thomas; Walheim, Stefan

    2013-01-01

    A detailed understanding of the interface between living cells and substrate materials is of rising importance in many fields of medicine, biology and biotechnology. Cells at interfaces often form epithelia. The physical barrier that they form is one of their main functions. It is governed by the properties of the networks forming the cytoskeleton systems and by cell-to-cell contacts. Different substrates with varying surface properties modify the migration velocity of the cells. On the one hand one can change the materials composition. Organic and inorganic materials induce differing migration velocities in the same cell system. Within the same class of materials, a change of the surface stiffness or of the surface energy modifies the migration velocity, too. For our cell adhesion studies a variety of different, homogeneous substrates were used (polymers, bio-polymers, metals, oxides). In addition, an effective lithographic method, Polymer Blend Lithography (PBL), is reported, to produce patterned Self-Assem...

  3. Yeast surface display of dehydrogenases in microbial fuel-cells.

    Science.gov (United States)

    Gal, Idan; Schlesinger, Orr; Amir, Liron; Alfonta, Lital

    2016-12-01

    Two dehydrogenases, cellobiose dehydrogenase from Corynascus thermophilus and pyranose dehydrogenase from Agaricus meleagris, were displayed for the first time on the surface of Saccharomyces cerevisiae using the yeast surface display system. Surface displayed dehydrogenases were used in a microbial fuel cell and generated high power outputs. Surface displayed cellobiose dehydrogenase has demonstrated a midpoint potential of -28mV (vs. Ag/AgCl) at pH=6.5 and was used in a mediator-less anode compartment of a microbial fuel cell producing a power output of 3.3μWcm(-2) using lactose as fuel. Surface-displayed pyranose dehydrogenase was used in a microbial fuel cell and generated high power outputs using different substrates, the highest power output that was achieved was 3.9μWcm(-2) using d-xylose. These results demonstrate that surface displayed cellobiose dehydrogenase and pyranose dehydrogenase may successfully be used in microbial bioelectrochemical systems.

  4. Yeast surface display of dehydrogenases in microbial fuel-cells.

    Science.gov (United States)

    Gal, Idan; Schlesinger, Orr; Amir, Liron; Alfonta, Lital

    2016-12-01

    Two dehydrogenases, cellobiose dehydrogenase from Corynascus thermophilus and pyranose dehydrogenase from Agaricus meleagris, were displayed for the first time on the surface of Saccharomyces cerevisiae using the yeast surface display system. Surface displayed dehydrogenases were used in a microbial fuel cell and generated high power outputs. Surface displayed cellobiose dehydrogenase has demonstrated a midpoint potential of -28mV (vs. Ag/AgCl) at pH=6.5 and was used in a mediator-less anode compartment of a microbial fuel cell producing a power output of 3.3μWcm(-2) using lactose as fuel. Surface-displayed pyranose dehydrogenase was used in a microbial fuel cell and generated high power outputs using different substrates, the highest power output that was achieved was 3.9μWcm(-2) using d-xylose. These results demonstrate that surface displayed cellobiose dehydrogenase and pyranose dehydrogenase may successfully be used in microbial bioelectrochemical systems. PMID:27459246

  5. Design and fabrication of miniaturized PEM fuel cell combined microreactor with self-regulated hydrogen mechanism

    Science.gov (United States)

    Balakrishnan, A.; Frei, M.; Kerzenmacher, S.; Reinecke, H.; Mueller, C.

    2015-12-01

    In this work we present the design and fabrication of the miniaturized PEM fuel cell combined microreactor system with hydrogen regulation mechanism and testing of prototype microreactor. The system consists of two components (i) fuel cell component and (ii) microreactor component. The fuel cell component represents the miniaturized PEM fuel cell system (combination of screen printed fuel cell assembly and an on-board hydrogen storage medium). Hydrogen production based on catalytic hydrolysis of chemical hydride takes place in the microreactor component. The self-regulated hydrogen mechanism based on the gaseous hydrogen produced from the catalytic hydrolysis of sodium borohydride (NaBH4) gets accumulated as bubbles at the vicinity of the hydrophobic coated hydrogen exhaust holes. When the built up hydrogen bubbles pressure exceeds the burst pressure at the hydrogen exhaust holes the bubble collapses. This collapse causes a surge of fresh NaBH4 solution onto the catalyst surface leading to the removal of the reaction by-products formed at the active sites of the catalyst. The catalyst used in the system is platinum deposited on a base substrate. Nickel foam, carbon porous medium (CPM) and ceramic plate were selected as candidates for base substrate for developing a robust catalyst surface. For the first time the platinum layer fabricated by pulsed electrodeposition and dealloying (EPDD) technique is used for hydrolysis of NaBH4. The major advantages of such platinum catalyst layers are its high surface area and their mechanical stability. Prototype microreactor system with self-regulated hydrogen mechanism is demonstrated.

  6. Surface complexation of neptunium (V) onto whole cells and cell componets of Shewanella alga

    Energy Technology Data Exchange (ETDEWEB)

    Reed, Donald Timothy [Los Alamos National Laboratory; Deo, Randhir P [ASU; Rittmann, Bruce E [ASU; Songkasiri, Warinthorn [UNAFFILIATED

    2008-01-01

    We systematically quantified surface complexation of neptunium(V) onto whole cells of Shewanella alga strain BrY and onto cell wall and extracellular polymeric substances (EPS) of S. alga. We first performed acid and base titrations and used the mathematical model FITEQL with constant-capacitance surface-complexation to determine the concentrations and deprotonation constants of specific surface functional groups. Deprotonation constants most likely corresponded to a carboxyl site associated with amino acids (pK{sub a} {approx} 2.4), a carboxyl group not associated with amino acids (pK{sub a} {approx} 5), a phosphoryl site (pK{sub a} {approx} 7.2), and an amine site (pK{sub a} > 10). We then carried out batch sorption experiments with Np(V) and each of the S. alga components at different pHs. Results show that solution pH influenced the speciation of Np(V) and each of the surface functional groups. We used the speciation sub-model of the biogeochemical model CCBATCH to compute the stability constants for Np(V) complexation to each surface functional group. The stability constants were similar for each functional group on S. alga bacterial whole cells, cell walls, and EPS, and they explain the complicated sorption patterns when they are combined with the aqueous-phase speciation of Np(V). For pH < 8, NpO{sub 2}{sup +} was the dominant form of Np(V), and its log K values for the low-pK{sub a} carboxyl, other carboxyl, and phosphoryl groups were 1.75, 1.75, and 2.5 to 3.1, respectively. For pH greater than 8, the key surface ligand was amine >XNH3+, which complexed with NpO{sub 2}(CO{sub 3}){sub 3}{sup 5-}. The log K for NpO{sub 2}(CO{sub 3}){sub 3}{sup 5-} complexed onto the amine groups was 3.1 to 3.6. All of the log K values are similar to those of Np(V) complexes with aqueous carboxyl and N-containing carboxyl ligands. These results point towards the important role of surface complexation in defining key actinide-microbiological interactions in the subsurface.

  7. Combined effects of surface conditions, boundary layer dynamics and chemistry on diurnal SOA evolution

    NARCIS (Netherlands)

    Janssen, R.H.H.; Vilà-Guerau de Arellano, J.; Ganzeveld, L.N.; Kabat, P.; Jimenez, J.L.; Farmer, D.K.; Heerwaarden, van C.C.; Mammarella, I.

    2012-01-01

    We study the combined effects of land surface conditions, atmospheric boundary layer dynamics and chemistry on the diurnal evolution of biogenic secondary organic aerosol in the atmospheric boundary layer, using a model that contains the essentials of all these components. First, we evaluate the mod

  8. Control of surface wettability for inkjet printing by combining hydrophobic coating and plasma treatment

    International Nuclear Information System (INIS)

    We have obtained a wide range of surface wettabilities of PI substrate for inkjet printing by combining hydrophobic solution coating and O2 or Ar plasma treatments. Experiments were conducted to investigate the variation in inkjet-printed dot diameters with different surface treatments. The change in chemical and physical characteristics of treated surfaces was evaluated using static contact angle measurements, field emission scanning electron microscopy, atomic force microscopy, and X-ray photoelectron spectroscopy. Only hydrophobic coated surface produces the smallest dot diameter and the largest contact angle. Dot diameter increases and contact angle decreases as the plasma treatment time increases. Since the removal of hydrophobic layer from the surface occurs due to the etching effect of O2 and Ar plasma during the plasma treatments, F/C ratio decreases with increasing the plasma treatment time. Surface roughness variations are also observed after plasma treatments. The ranges of printed dot sizes for O2 and Ar plasma treatments are 38 μm–70 μm and 38 μm–92 μm, respectively. Ar plasma treatment shows a wider range of surface wettability because of higher removal rate of the hydrophobic layer. This combination of hydrophobic coating and plasma treatment can offer an effective way to obtain a wide range of surface wettabilities for high quality inkjet-printed patterns. - Highlights: • Hydrophobic coating and plasma treatments were used to control surface wettability. • Inkjet-printed dot diameters increase with O2 or Ar plasma treatment time. • Contact angles of Ag ink agree well with the variation tendency of dot diameters. • The removal of hydrophobic layer occurs during the plasma treatments. • Ar plasma treatment shows a wider range of surface wettability than O2 plasma

  9. One Step Quick Detection of Cancer Cell Surface Marker by Integrated NiFe-based Magnetic Biosensing Cell Cultural Chip

    Institute of Scientific and Technical Information of China (English)

    Chenchen Bao; Lei Chen; Tao Wang; Chong Lei; Furong Tian; Daxiang Cui; Yong Zhou

    2013-01-01

    RGD peptides has been used to detect cell surface integrin and direct clinical effective therapeutic drug selection. Herein we report that a quick one step detection of cell surface marker that was realized by a specially designed NiFe-based magnetic biosensing cell chip combined with functionalized magnetic nanoparti-cles. Magnetic nanoparticles with 20-30 nm in diameter were prepared by coprecipitation and modified with RGD-4C, and the resultant RGD-functionalized magnetic nanoparticles were used for targeting cancer cells cul-tured on the NiFe-based magnetic biosensing chip and distinguish the amount of cell surface receptor-integrin. Cell lines such as Calu3, Hela, A549, CaFbr, HEK293 and HUVEC exhibiting different integrin expression were chosen as test samples. Calu3, Hela, HEK293 and HUVEC cells were successfully identified. This approach has advantages in the qualitative screening test. Compared with traditional method, it is fast, sensitive, low cost, easy-operative, and needs very little human intervention. The novel method has great potential in applications such as fast clinical cell surface marker detection, and diagnosis of early cancer, and can be easily extended to other biomedical applications based on molecular recognition.

  10. Surface Passivation Studies on n+pp+ Bifacial Solar Cell

    OpenAIRE

    Suhaila Sepeai; M. Y. Sulaiman; Kamaruzzaman Sopian; Saleem H. Zaidi

    2012-01-01

    Bifacial solar cell is a specially designed solar cell for the production of electricity from both sides of the solar cell. It is an active field of research to make photovoltaics (PV) more competitive by increasing its efficiency and lowering its costs. We developed an n+pp+ structure for the bifacial solar cell. The fabrication used phosphorus-oxy-trichloride (POCl3) diffusion to form the emitter and Al diffusion using conventional screen printing to produce the back surface field (BSF). Th...

  11. Interaction between antitumor drug and silver nanoparticles:combined fluorscence and surface enhanced Raman scattering study

    Institute of Scientific and Technical Information of China (English)

    Jing Yang; Hong Wang; Zhuyuan Wang; Xuebin Tan; Chunyuan Song; Ruohu Zhang; Jin Li; Yiping Cui

    2009-01-01

    Optical methods and MTT method are used to characterize the antiproliferation effect of antitumor drug 9-aminoacridine (9AA) with and without silver nanoparticles.Intracellular surface enhanced Raman scat tering (SERS) spectra and fluorescent spectra of 9AA indicate the form of 9AA adsorbed on the surface of silver nanoparticles.Although both silver nanoparticles and antitumor drug can inhibit the growth of Hela cells,silver nanoparticles can slow down the antiproliferation effect on Hela cells at low concentration of antitumor drugs.Our experimental results suggest that silver nanoparticles may serve as slow-release drug carriers,which is important in antitumor drug delivery.

  12. Letter to the EditorRetrieval of land surface temperature from combined AVHRR data

    Directory of Open Access Journals (Sweden)

    H. Fischer

    Full Text Available Accurate retrievals of land surface temperature (LST from space are of high interest for studies of land surface processes. Here, an operationally applicable method to retrieve LST from NOAA/AVHRR data is proposed, which combines a split-window technique (SWT for atmospheric correction with a Normalised Difference Vegetation Index threshold method for the retrieval of land surface emissivity. Preliminary results of LST retrievals with this "combined method" are in good agreement with ground truth measurements for bare soil and wheat crops. The results are also compared with results from the same SWT but using emissivities from laboratory measurements.Key words. Meteorology and atmospheric dynamics (radiation processes; instruments and techniques – Radio science (remote sensing

  13. Hardening of the surface layers of commercial pure titanium VT1-0 under combined treatment

    International Nuclear Information System (INIS)

    The treatment of VT1-0 titanium samples was carried out by concentrated energy fluxes. The combined treatment included surface carburizing with the joint use of powder samples of compounds with high physical and mechanical properties (namely, titanium diboride TiB2, silicon carbide SiC and zirconium oxide ZrO2) and subsequent electron beam treatment of surface layers formed in electroexplosive treatment. The combined treatment of surface layers resulted in the multifold increase in microhardness, which reduces depending on the depth of hardening zone. After electron-beam treatment, the depth of hardening zone is increased. During electron-beam treatment, the two-layer hardening zone forms

  14. Hardening of the surface layers of commercial pure titanium VT1-0 under combined treatment

    Energy Technology Data Exchange (ETDEWEB)

    Bashchenko, Lyudmila P., E-mail: luda.baschenko@gmail.com; Gromov, Viktor E., E-mail: gromov@physics.sibsiu.ru; Budovskikh, Evgenii A., E-mail: budovskih-ea@physics.sibsiu.ru; Soskova, Nina A., E-mail: gromov@physics.sibsiu.ru [Siberian State Industrial University, Novokuznetsk, 654007 (Russian Federation); Ivanov, Yurii F., E-mail: yufi55@mail.ru [Institute of High Current Electronics SB RAS, Tomsk, 634055 (Russian Federation); National Research Tomsk State University, Tomsk, 634050 (Russian Federation)

    2015-10-27

    The treatment of VT1-0 titanium samples was carried out by concentrated energy fluxes. The combined treatment included surface carburizing with the joint use of powder samples of compounds with high physical and mechanical properties (namely, titanium diboride TiB{sub 2}, silicon carbide SiC and zirconium oxide ZrO{sub 2}) and subsequent electron beam treatment of surface layers formed in electroexplosive treatment. The combined treatment of surface layers resulted in the multifold increase in microhardness, which reduces depending on the depth of hardening zone. After electron-beam treatment, the depth of hardening zone is increased. During electron-beam treatment, the two-layer hardening zone forms.

  15. Hardening of the surface layers of commercial pure titanium VT1-0 under combined treatment

    Science.gov (United States)

    Bashchenko, Lyudmila P.; Gromov, Viktor E.; Budovskikh, Evgenii A.; Ivanov, Yurii F.; Soskova, Nina A.

    2015-10-01

    The treatment of VT1-0 titanium samples was carried out by concentrated energy fluxes. The combined treatment included surface carburizing with the joint use of powder samples of compounds with high physical and mechanical properties (namely, titanium diboride TiB2, silicon carbide SiC and zirconium oxide ZrO2) and subsequent electron beam treatment of surface layers formed in electroexplosive treatment. The combined treatment of surface layers resulted in the multifold increase in microhardness, which reduces depending on the depth of hardening zone. After electron-beam treatment, the depth of hardening zone is increased. During electron-beam treatment, the two-layer hardening zone forms.

  16. Ancestral vascular lumen formation via basal cell surfaces.

    Directory of Open Access Journals (Sweden)

    Tomás Kucera

    Full Text Available The cardiovascular system of bilaterians developed from a common ancestor. However, no endothelial cells exist in invertebrates demonstrating that primitive cardiovascular tubes do not require this vertebrate-specific cell type in order to form. This raises the question of how cardiovascular tubes form in invertebrates? Here we discovered that in the invertebrate cephalochordate amphioxus, the basement membranes of endoderm and mesoderm line the lumen of the major vessels, namely aorta and heart. During amphioxus development a laminin-containing extracellular matrix (ECM was found to fill the space between the basal cell surfaces of endoderm and mesoderm along their anterior-posterior (A-P axes. Blood cells appear in this ECM-filled tubular space, coincident with the development of a vascular lumen. To get insight into the underlying cellular mechanism, we induced vessels in vitro with a cell polarity similar to the vessels of amphioxus. We show that basal cell surfaces can form a vascular lumen filled with ECM, and that phagocytotic blood cells can clear this luminal ECM to generate a patent vascular lumen. Therefore, our experiments suggest a mechanism of blood vessel formation via basal cell surfaces in amphioxus and possibly in other invertebrates that do not have any endothelial cells. In addition, a comparison between amphioxus and mouse shows that endothelial cells physically separate the basement membranes from the vascular lumen, suggesting that endothelial cells create cardiovascular tubes with a cell polarity of epithelial tubes in vertebrates and mammals.

  17. Nanoscale crystallinity modulates cell proliferation on plasma sprayed surfaces

    Energy Technology Data Exchange (ETDEWEB)

    Smith, Alan M. [School of Applied Sciences, University of Huddersfield, Huddersfield HD1 3DH (United Kingdom); Paxton, Jennifer Z.; Hung, Yi-Pei; Hadley, Martin J.; Bowen, James; Williams, Richard L. [School of Chemical Engineering, University of Birmingham, Edgbaston, B15 2TT (United Kingdom); Grover, Liam M., E-mail: l.m.grover@bham.ac.uk [School of Chemical Engineering, University of Birmingham, Edgbaston, B15 2TT (United Kingdom)

    2015-03-01

    Calcium phosphate coatings have been applied to the surface of metallic prostheses to mediate hard and soft tissue attachment for more than 40 years. Most coatings are formed of high purity hydroxyapatite, and coating methods are often designed to produce highly crystalline surfaces. It is likely however, that coatings of lower crystallinity can facilitate more rapid tissue attachment since the surface will exhibit a higher specific surface area and will be considerably more reactive than a comparable highly crystalline surface. Here we test this hypothesis by growing a population of MC3T3 osteoblast-like cells on the surface of two types of hip prosthesis with similar composition, but with differing crystallinity. The surfaces with lower crystallinity facilitated more rapid cell attachment and increased proliferation rate, despite having a less heterogeneous surface topography. This work highlights that the influence of the crystallinity of HA at the nano-scale is dominant over macro-scale topography for cell adhesion and growth. Furthermore, crystallinity could be easily adjusted by without compromising coating purity. These findings could facilitate designing novel coated calcium phosphate surfaces that more rapidly bond tissue following implantation. - Highlights: • Crystallinity of HA at the nano-scale was dominant over macro-scale topography. • Lower crystallinity caused rapid cell attachment and proliferation rate. • Crystallinity could be easily adjusted by without compromising coating purity.

  18. Oligonucleotide delivery with cell surface binding and cell penetrating Peptide amphiphile nanospheres.

    Science.gov (United States)

    Mumcuoglu, Didem; Sardan, Melis; Tekinay, Turgay; Guler, Mustafa O; Tekinay, Ayse B

    2015-05-01

    A drug delivery system designed specifically for oligonucleotide therapeutics can ameliorate the problems associated with the in vivo delivery of these molecules. The internalization of free oligonucleotides is challenging, and cytotoxicity is the main obstacle for current transfection vehicles. To develop nontoxic delivery vehicles for efficient transfection of oligonucleotides, we designed a self-assembling peptide amphiphile (PA) nanosphere delivery system decorated with cell penetrating peptides (CPPs) containing multiple arginine residues (R4 and R8), and a cell surface binding peptide (KRSR), and report the efficiency of this system in delivering G-3129, a Bcl-2 antisense oligonucleotide (AON). PA/AON (peptide amphiphile/antisense oligonucleotide) complexes were characterized with regards to their size and secondary structure, and their cellular internalization efficiencies were evaluated. The effect of the number of arginine residues on the cellular internalization was investigated by both flow cytometry and confocal imaging, and the results revealed that uptake efficiency improved as the number of arginines in the sequence increased. The combined effect of cell penetration and surface binding property on the cellular internalization and its uptake mechanism was also evaluated by mixing R8-PA and KRSR-PA. R8 and R8/KRSR decorated PAs were found to drastically increase the internalization of AONs compared to nonbioactive PA control. Overall, the KRSR-decorated self-assembled PA nanospheres were demonstrated to be noncytotoxic delivery vectors with high transfection rates and may serve as a promising delivery system for AONs. PMID:25828697

  19. Cell surface engineering of industrial microorganisms for biorefining applications.

    Science.gov (United States)

    Tanaka, Tsutomu; Kondo, Akihiko

    2015-11-15

    In order to decrease carbon emissions and negative environmental impacts of various pollutants, biofuel/biochemical production should be promoted for replacing fossil-based industrial processes. Utilization of abundant lignocellulosic biomass as a feedstock has recently become an attractive option. In this review, we focus on recent efforts of cell surface display using industrial microorganisms such as Escherichia coli and yeast. Cell surface display is used primarily for endowing cellulolytic activity on the host cells, and enables direct fermentation to generate useful fuels and chemicals from lignocellulosic biomass. Cell surface display systems are systematically summarized, and the drawbacks/perspectives as well as successful application of surface display for industrial biotechnology are discussed.

  20. Cell surface engineering of industrial microorganisms for biorefining applications.

    Science.gov (United States)

    Tanaka, Tsutomu; Kondo, Akihiko

    2015-11-15

    In order to decrease carbon emissions and negative environmental impacts of various pollutants, biofuel/biochemical production should be promoted for replacing fossil-based industrial processes. Utilization of abundant lignocellulosic biomass as a feedstock has recently become an attractive option. In this review, we focus on recent efforts of cell surface display using industrial microorganisms such as Escherichia coli and yeast. Cell surface display is used primarily for endowing cellulolytic activity on the host cells, and enables direct fermentation to generate useful fuels and chemicals from lignocellulosic biomass. Cell surface display systems are systematically summarized, and the drawbacks/perspectives as well as successful application of surface display for industrial biotechnology are discussed. PMID:26070720

  1. Cell surface carbohydrates as prognostic markers in human carcinomas

    DEFF Research Database (Denmark)

    Dabelsteen, Erik

    1996-01-01

    Tumour development is usually associated with changes in cell surface carbohydrates. These are often divided into changes related to terminal carbohydrate structures, which include incomplete synthesis and modification of normally existing carbohydrates, and changes in the carbohydrate core struc...

  2. Precise manipulation of cell behaviors on surfaces for construction of tissue/organs.

    Science.gov (United States)

    Zheng, Wenfu; Jiang, Xingyu

    2014-12-01

    The use of micro/nanotechnology has become an indispensable strategy to manipulating cell microenvironments. By employing key elements of soft lithographical technologies including self-assembled monolayers (SAMs), microcontact printing (μCP), and microfluidic pattering (μFP) and a number of switchable surfaces such as electrochemical active, photosensitive, and thermosensitive surfaces, scientists can control the adhesion, proliferation, migration and differentiation of cells. By combining essential in vivo conditions, various physical or pathological processes such as cell-cell interaction in wound healing and tumor metastasis could be studied on well-defined surfaces and interfaces. By integrating key elements in live tissues, in vitro models mimicking basic structure and function of vital organs such as lung, heart, blood vessel, liver, kidney, and brain have been developed and greatly increased our knowledge of these important life processes. In this review, we will focus on the recent development of these interfacial methods and their application in fundamental biology research.

  3. Surface texturing of multicrystalline silicon solar cells

    OpenAIRE

    L.A. Dobrzański; A. Drygała

    2008-01-01

    Purpose: The aim of the paper is to elaborate a laser method of texturization multicrystalline silicon. The main reason for taking up the research is that most conventional methods used for texturization of monocrystalline silicon are ineffective when applied for texturing multicrystalline silicon. This is related to random distribution of grains of different crystalographic orientations on the surface of multicrystalline silicon.Design/methodology/approach: The topography of laser ...

  4. Spatial resolution in thin film deposition on silicon surfaces by combining silylation and UV/ozonolysis

    Science.gov (United States)

    Guo, Lei; Zaera, Francisco

    2014-12-01

    A simple procedure has been developed for the processing of silicon wafers in order to facilitate the spatially resolved growth of thin solid films on their surfaces. Specifically, a combination of silylation and UV/ozonolysis was tested as a way to control the concentration of the surface hydroxo groups required for subsequent atomic layer deposition (ALD) of metals or oxides. Water contact angle measurements were used to evaluate the hydrophilicity/hydrophobicity of the surface, a proxy for OH surface coverage, and to optimize the UV/ozonolysis treatment. Silylation with hexamethyldisilazane, trichloro(octadecyl)silane, or trimethylchlorosilane was found to be an efficient way to block the hydroxo sites and to passivate the underlying surface, and UV/O3 treatments were shown to effectively remove the silylation layer and to regain the surface reactivity. Both O3 and 185 nm UV radiation were determined necessary for the removal of the silylation layer, and additional 254 nm radiation was found to enhance the process. Attenuated total reflection-infrared absorption spectroscopy was employed to assess the success of the silylation and UV/O3 removal steps, and atomic force microscopy data provided evidence for the retention of the original smoothness of the surface. Selective growth of HfO2 films via TDMAHf + H2O ALD was seen only on the UV/O3 treated surfaces; total inhibition of the deposition was observed on the untreated silylated surfaces (as determined by x-ray photoelectron spectroscopy and ellipsometry). Residual film growth was still detected on the latter if the ALD was carried out at high temperatures (250 °C), because the silylation layer deteriorates under such harsh conditions and forms surface defects that act as nucleation sites for the growth of oxide grains (as identified by electron microscopy and scanning electron microscopy). We believe that the silylation-UV/O3 procedure advanced here could be easily implemented for the patterning of surfaces

  5. Recent Insights into Cell Surface Heparan Sulphate Proteoglycans and Cancer.

    Science.gov (United States)

    Couchman, John R; Multhaupt, Hinke; Sanderson, Ralph D

    2016-01-01

    A small group of cell surface receptors are proteoglycans, possessing a core protein with one or more covalently attached glycosaminoglycan chains. They are virtually ubiquitous and their chains are major sites at which protein ligands of many types interact. These proteoglycans can signal and regulate important cell processes, such as adhesion, migration, proliferation, and differentiation. Since many protein ligands, such as growth factors, morphogens, and cytokines, are also implicated in tumour progression, it is increasingly apparent that cell surface proteoglycans impact tumour cell behaviour. Here, we review some recent advances, emphasising that many tumour-related functions of proteoglycans are revealed only after their modification in processes subsequent to synthesis and export to the cell surface. These include enzymes that modify heparan sulphate structure, recycling of whole or fragmented proteoglycans into exosomes that can be paracrine effectors or biomarkers, and lateral interactions between some proteoglycans and calcium channels that impact the actin cytoskeleton. PMID:27408707

  6. Recent Insights into Cell Surface Heparan Sulphate Proteoglycans and Cancer

    Science.gov (United States)

    Couchman, John R; Multhaupt, Hinke; Sanderson, Ralph D.

    2016-01-01

    A small group of cell surface receptors are proteoglycans, possessing a core protein with one or more covalently attached glycosaminoglycan chains. They are virtually ubiquitous and their chains are major sites at which protein ligands of many types interact. These proteoglycans can signal and regulate important cell processes, such as adhesion, migration, proliferation, and differentiation. Since many protein ligands, such as growth factors, morphogens, and cytokines, are also implicated in tumour progression, it is increasingly apparent that cell surface proteoglycans impact tumour cell behaviour. Here, we review some recent advances, emphasising that many tumour-related functions of proteoglycans are revealed only after their modification in processes subsequent to synthesis and export to the cell surface. These include enzymes that modify heparan sulphate structure, recycling of whole or fragmented proteoglycans into exosomes that can be paracrine effectors or biomarkers, and lateral interactions between some proteoglycans and calcium channels that impact the actin cytoskeleton. PMID:27408707

  7. Cell surface localization and tissue distribution of a hepatocyte cell-cell adhesion glycoprotein (cell-CAM 105)

    OpenAIRE

    Ocklind, C; Forsum, U; Obrink, B

    1983-01-01

    We recently identified a 105,000-dalton plasma membrane glycoprotein, denoted cell-CAM 105 (CAM, cell adhesion molecule), that is involved in intercellular adhesion of reaggregating rat hepatocytes (Ocklind, C., and B. Obrink, 1982, J. Biol. Chem., 257:6788-6795). In this communication we used a monospecific rabbit antiserum against cell-CAM 105 to localize the antigen by indirect immunofluorescence on isolated rat cells and on frozen rat tissue sections. This antiserum stained the surface of...

  8. Immunogold labels: cell-surface markers in atomic force microscopy

    NARCIS (Netherlands)

    Putman, Constant A.J.; Grooth, de Bart G.; Hansma, Paul K.; Hulst, van Niek F.; Greve, Jan

    1993-01-01

    The feasibility of using immunogold labels as cell-surface markers in atomic force microscopy is shown in this paper. The atomic force microscope (AFM) was used to image the surface of immunogold-labeled human lymphocytes. The lymphocytes were isolated from whole blood and labeled by an indirect imm

  9. Nanometer polymer surface features: the influence on surface energy, protein adsorption and endothelial cell adhesion

    Science.gov (United States)

    Carpenter, Joseph; Khang, Dongwoo; Webster, Thomas J.

    2008-12-01

    Current small diameter (vascular graft materials exhibit poor long-term patency due to thrombosis and intimal hyperplasia. Tissue engineered solutions have yielded functional vascular tissue, but some require an eight-week in vitro culture period prior to implantation—too long for immediate clinical bedside applications. Previous in vitro studies have shown that nanostructured poly(lactic-co-glycolic acid) (PLGA) surfaces elevated endothelial cell adhesion, proliferation, and extracellular matrix synthesis when compared to nanosmooth surfaces. Nonetheless, these studies failed to address the importance of lateral and vertical surface feature dimensionality coupled with surface free energy; nor did such studies elicit an optimum specific surface feature size for promoting endothelial cell adhesion. In this study, a series of highly ordered nanometer to submicron structured PLGA surfaces of identical chemistry were created using a technique employing polystyrene nanobeads and poly(dimethylsiloxane) (PDMS) molds. Results demonstrated increased endothelial cell adhesion on PLGA surfaces with vertical surface features of size less than 18.87 nm but greater than 0 nm due to increased surface energy and subsequently protein (fibronectin and collagen type IV) adsorption. Furthermore, this study provided evidence that the vertical dimension of nanometer surface features, rather than the lateral dimension, is largely responsible for these increases. In this manner, this study provides key design parameters that may promote vascular graft efficacy.

  10. Expanding the diversity of unnatural cell surface sialic acids

    Energy Technology Data Exchange (ETDEWEB)

    Luchansky, Sarah J.; Goon, Scarlett; Bertozzi, Carolyn R.

    2003-10-30

    Novel chemical reactivity can be introduced onto cell surfaces through metabolic oligosaccharide engineering. This technique exploits the substrate promiscuity of cellular biosynthetic enzymes to deliver unnatural monosaccharides bearing bioorthogonal functional groups into cellular glycans. For example, derivatives of N-acetylmannosamine (ManNAc) are converted by the cellular biosynthetic machinery into the corresponding sialic acids and subsequently delivered to the cell surface in the form of sialoglycoconjugates. Analogs of N-acetylglucosamine (GlcNAc) and N-acetylgalactosamine (GalNAc) are also metabolized and incorporated into cell surface glycans, likely through the sialic acid and GalNAc salvage pathways, respectively. Furthermore, GlcNAc analogs can be incorporated into nucleocytoplasmic proteins in place of {beta}-O-GlcNAc residues. These pathways have been exploited to integrate unique electrophiles such as ketones and azides into the target glycoconjugate class. These functional groups can be further elaborated in a chemoselective fashion by condensation with hydrazides and by Staudinger ligation, respectively, thereby introducing detectable probes onto the cell. In conclusion, sialic acid derivatives are efficient vehicles for delivery of bulky functional groups to cell surfaces and masking of their hydroxyl groups improves their cellular uptake and utilization. Furthermore, the successful introduction of photoactivatable aryl azides into cell surface glycans opens up new avenues for studying sialic acid-binding proteins and elucidating the role of sialic acid in essential processes such as signaling and cell adhesion.

  11. Characterization of the osteogenic potential of mesenchymal stem cells from human periodontal ligament based on cell surface markers

    Institute of Scientific and Technical Information of China (English)

    Ruth Alvarez; Hye-Lim Lee; Cun-Yu Wang; Christine Hong

    2015-01-01

    Mesenchymal stem cell (MSC)-mediated therapy has been shown to be clinically effective in regenerating tissue defects. For improved regenerative therapy, it is critical to isolate homogenous populations of MSCs with high capacity to differentiate into appropriate tissues. The utilization of stem cell surface antigens provides a means to identify MSCs from various tissues. However, few surface markers that consistently isolate highly regenerative MSCs have been validated, making it challenging for routine clinical applications and making it all the more imperative to identify reliable surface markers. In this study, we used three surface marker combinations:CD51/CD140a, CD271, and STRO-1/CD146 for the isolation of homogenous populations of dental mesenchymal stem cells (DMSCs) from heterogeneous periodontal ligament cells (PDLCs). Fluorescence-activated cell sorting analysis revealed that 24%of PDLCs were CD511/CD140a1, 0.8%were CD2711, and 2.4%were STRO-11/CD1461. Sorted cell populations were further assessed for their multipotent properties by inducing osteogenic and chondrogenic differentiation. All three subsets of isolated DMSCs exhibited differentiation capacity into osteogenic and chondrogenic lineages but with varying degrees. CD2711 DMSCs demonstrated the greatest osteogenic potential with strong induction of osteogenic markers such as DLX5, RUNX2, and BGLAP. Our study provides evidence that surface marker combinations used in this study are sufficient markers for the isolation of DMSCs from PDLCs. These results provide important insight into using specific surface markers for identifying homogenous populations of DMSCs for their improved utilization in regenerative medicine.

  12. Study of surface cell Madelung constant and surface free energy of nanosized crystal grain

    Institute of Scientific and Technical Information of China (English)

    Zhang Wei-Jia; Wang Tian-Min; Rong Ai-Lun; Cui Min

    2006-01-01

    Surface cell Madelung constant is firstly defined for calculating the surface free energy of nanosized crystal grains,which explains the physical performance of small crystals and may be greatly beneficial to the analysis of surface states and the study of the dynamics of crystal nucleation and growth.A new approximative expression of the surface energy and relevant thermodynamic data are used in this calculation.New formula and computing method for calculating the Madelung constant α of any complex crystals are proposed,and the surface free energies and surface electrostatic energies of nanosized crystal grains and the Madelung constant of some complex crystals are theoretically calculated in this paper.The surface free energy of nanosized-crystal-grain TiO2 and the surface electrostatic energy (absolute value) of nanosized-crystal-grain α-A12O3 are found to be the biggest among all the crystal grains including those of other species.

  13. Study of Surface Cell Madelung Constant and Surface Free Energy of Nanosized Crystal Grain

    Institute of Scientific and Technical Information of China (English)

    ZHANG Wei-Jia; WANG Tian-Min; CUI Min

    2005-01-01

    Surface cell Madelung constant is firstly defined in calculating surface free energy of nanosized crystal grains, which explains the physical performance of small crystals and may be great benefit to make surface analysis and study dynamics of crystal nucleus growth. A new ap- proximative expression of surface energy and relevant thermodynamic data was used in this cal- culation. A new formula and computing method for calculating the Madelung constant α of any complex crystals is proposed, and surface free energies and surface electrostatic energies of nano- sized crystal grains as well as Madelung constant of some complex crystals are theoretically cal- culated in this paper. The surface free energy of nanosized crystal grain TiO2 and surface elec- trostatic energy(absolute value) of nanosized crystal grain α-Al2O3 are found to be the biggest among other crystal grains.

  14. Improved accuracy of cell surface shaving proteomics in Staphylococcus aureus using a false-positive control

    DEFF Research Database (Denmark)

    Solis, Nestor; Larsen, Martin Røssel; Cordwell, Stuart J

    2010-01-01

    with either trypsin or proteinase-K combined with LC-MS/MS. Trypsin-derived data were controlled using a "false-positive" strategy where cells were incubated without protease, removed by centrifugation and the resulting supernatants digested. Peptides identified in this fraction most likely result from cell...... lysis and were removed from the trypsin-shaved data set. We identified 42 predicted S. aureus COL surface proteins from 260 surface-exposed peptides. Trypsin and proteinase-K digests were highly complementary with ten proteins identified by both, 16 specific to proteinase-K treatment, 13 specific...

  15. Formation of Combined Surface Features of Protrusion Array and Wrinkles atop Shape-Memory Polymer

    Science.gov (United States)

    Sun, L.; Zhao, Y.; Huang, W. M.; Tong, T. H.

    We demonstrate a simple and cost-effective approach to realize two combined surface features of different scales together, namely submillimeter-sized protrusion array and microwrinkles, atop a polystyrene shape-memory polymer. Two different types of protrusions, namely flat-top protrusion and crown-shaped protrusion, were studied. The array of protrusions was produced by the Indentation-Polishing-Heating (IPH) process. Compactly packed steel balls were used for making array of indents. A thin gold layer was sputter deposited atop the polymer surface right after polishing. After heating for shape recovery, array of protrusions with wrinkles on the top due to the buckling of gold layer was produced.

  16. Amplified effect of surface charge on cell adhesion by nanostructures

    Science.gov (United States)

    Xu, Li-Ping; Meng, Jingxin; Zhang, Shuaitao; Ma, Xinlei; Wang, Shutao

    2016-06-01

    Nano-biointerfaces with varied surface charge can be readily fabricated by integrating a template-based process with maleimide-thiol coupling chemistry. Significantly, nanostructures are employed for amplifying the effect of surface charge on cell adhesion, as revealed by the cell-adhesion performance, cell morphology and corresponding cytoskeletal organization. This study may provide a promising strategy for developing new biomedical materials with tailored cell adhesion for tissue implantation and regeneration.Nano-biointerfaces with varied surface charge can be readily fabricated by integrating a template-based process with maleimide-thiol coupling chemistry. Significantly, nanostructures are employed for amplifying the effect of surface charge on cell adhesion, as revealed by the cell-adhesion performance, cell morphology and corresponding cytoskeletal organization. This study may provide a promising strategy for developing new biomedical materials with tailored cell adhesion for tissue implantation and regeneration. Electronic supplementary information (ESI) available: Experimental details, SEM, KFM AFM, chemical modification and characterization. See DOI: 10.1039/c6nr00649c

  17. A naringenin–tamoxifen combination impairs cell proliferation and survival of MCF-7 breast cancer cells

    Energy Technology Data Exchange (ETDEWEB)

    Hatkevich, Talia; Ramos, Joseph; Santos-Sanchez, Idalys; Patel, Yashomati M., E-mail: ympatel@uncg.edu

    2014-10-01

    Since over 60% of breast cancers are estrogen receptor positive (ER+), many therapies have targeted the ER. The ER is activated by both estrogen binding and phosphorylation. While anti-estrogen therapies, such as tamoxifen (Tam) have been successful they do not target the growth factor promoting phosphorylation of the ER. Other proliferation pathways such as the phosphatidylinositol-3 kinase, (PI3K) and the mitogen-activated protein kinase (MAPK) pathways are activated in breast cancer cells and are associated with poor prognosis. Thus targeting multiple cellular proliferation and survival pathways at the onset of treatment is critical for the development of more effective therapies. The grapefruit flavanone naringenin (Nar) is an inhibitor of both the PI3K and MAPK pathways. Previous studies examining either Nar or Tam used charcoal-stripped serum which removed estrogen as well as other factors. We wanted to use serum containing medium in order to retain all the potential inducers of cell proliferation so as not to exclude any targets of Nar. Here we show that a Nar–Tam combination is more effective than either Tam alone or Nar alone in MCF-7 breast cancer cells. We demonstrate that a Nar–Tam combination impaired cellular proliferation and viability to a greater extent than either component alone in MCF-7 cells. Furthermore, the use of a Nar–Tam combination requires lower concentrations of both compounds to achieve the same effects on proliferation and viability. Nar may function by inhibiting both PI3K and MAPK pathways as well as localizing ERα to the cytoplasm in MCF-7 cells. Our results demonstrate that a Nar–Tam combination induces apoptosis and impairs proliferation signaling to a greater extent than either compound alone. These studies provide critical information for understanding the molecular mechanisms involved in cell proliferation and apoptosis in breast cancer cells. - Highlights: • Nar–Tam impairs cell viability more effectively than

  18. Biomimetic surface modification of titanium surfaces for early cell capture by advanced electrospinning

    International Nuclear Information System (INIS)

    The time required for osseointegration with a metal implant having a smooth surface ranges from three to six months. We hypothesized that biomimetic coating surfaces with poly(lactic-co-glycolic acid) (PLGA)/collagen fibers and nano-hydroxyapatite (n-HA) on the implant would enhance the adhesion of mesenchymal stem cells. Therefore, this surface modification of dental and bone implants might enhance the process of osseointegration. In this study, we coated PLGA or PLGA/collagen (50:50 w/w ratio) fiber on Ti disks by modified electrospinning for 5 s to 2 min; after that, we further deposited n-HA on the fibers. PLGA fibers of fiber diameter 0.957 ± 0.357 µm had a contact angle of 9.9 ± 0.3° and PLGA/collagen fibers of fiber diameter 0.378 ± 0.068 µm had a contact angle of 0°. Upon n-HA incorporation, all the fibers had a contact angle of 0° owing to the hydrophilic nature of n-HA biomolecule. The cell attachment efficiency was tested on all the scaffolds for different intervals of time (10, 20, 30 and 60 min). The alkaline phosphatase activity, cell proliferation and mineralization were analyzed on all the implant surfaces on days 7, 14 and 21. Results of the cell adhesion study indicated that the cell adhesion was maximum on the implant surface coated with PLGA/collagen fibers deposited with n-HA compared to the other scaffolds. Within a short span of 60 min, 75% of the cells adhered onto the mineralized PLGA/collagen fibers. Similarly by day 21, the rate of cell proliferation was significantly higher (p ≤ 0.05) on the mineralized PLGA/collagen fibers owing to enhanced cell adhesion on these fibers. This enhanced initial cell adhesion favored higher cell proliferation, differentiation and mineralization on the implant surface coated with mineralized PLGA/collagen fibers.

  19. Overexpression of Cell Surface Cytokeratin 8 in Multidrug-Resistant MCF-7/MX Cells Enhances Cell Adhesion to the Extracellular Matrix

    Directory of Open Access Journals (Sweden)

    Fang Liu

    2008-11-01

    Full Text Available Accumulating evidence suggests that multiple complex mechanisms may be involved, simultaneously or complementarily, in the emergence and development of multidrug resistance (MDR in various cancers. Cell adhesion-mediated MDR is one such mechanism. In the present study, we initially observed increased cell adhesion to extracellular matrix proteins by the MDR human breast tumor cell line MCF-7/MX compared to its parental cells. We then used a strategy that combined antibody-based screening technique and mass spectrometry-based proteomics to identify membrane proteins that contribute to the enhanced adhesion of MCF-7/MX cells. Using MCF-7/MX cells as immunogen, we isolated a mouse monoclonal antibody, 9C6, that preferentially reacts with MCF-7/MX cells over the parental MCF-7 cells. The molecular target of 9C6 was identified as cytokeratin 8 (CK8, which was found to be overexpressed on the cell surface of MCF-7/MX cells. We further observed that down-regulation of cell surface levels of CK8 through siRNA transfection significantly inhibited MCF-7/MX cell adhesion to fibronectin and vitronectin. In addition, anti-CK8 siRNA partially reversed the MDR phenotype of MCF-7/MX cells. Taken together, our results suggest that alterations in the expression level and cellular localization of CK8 may play a significant role in enhancing the cellular adhesion of MDR MCF-7/MX cells.

  20. Surface strategies for control of neuronal cell adhesion: A review

    Science.gov (United States)

    Roach, P.; Parker, T.; Gadegaard, N.; Alexander, M. R.

    2010-06-01

    Material engineering methods have been used for many years to develop biomedical devices for use within the body to augment, repair or replace damaged tissues ranging from contact lenses to heart valves. Here we review the findings gathered from the wide and varied surface analytical approaches applied to study the interaction between biology and man-made materials. The key material characteristics identified to be important for biological recognition are surface chemistry, topography and compliance. Model surfaces with controlled chemistry and topography have provided insight into biological response to various types of topographical features over a wide range of length scales from nano to micrometres, along with 3D matrices that have been used as scaffolds to support cells for tissue formation. The cellular response to surfaces with localised areas of patterned chemistry and to those presenting gradually changing chemistry are discussed. Where previous reviews have been structured around specific classes of surface modification, e.g. self-assembly, or have broadly examined the response of various cells to numerous surfaces, we aim in this article to focus in particular on the tissues involved in the nervous system whilst providing a broad overview of key issues from the field of cell and protein surface interactions with surfaces. The goal of repair and treatment of diseases related to the central and peripheral nervous systems rely on understanding the local interfacial environment and controlling responses at the cellular level. The role of the protein layer deposited from serum containing media onto man-made surfaces is discussed. We highlight the particular problems associated with the repair of the nervous system, and review how neuronal attachment and axon guidance can be accomplished using various surface cues when cultured with single and multiple cell types. We include a brief glossary of techniques discussed in the body of this article aimed at the

  1. Removal of diclofenac from surface water by electron beam irradiation combined with a biological aerated filter

    Science.gov (United States)

    He, Shijun; Wang, Jianlong; Ye, Longfei; Zhang, Youxue; Yu, Jiang

    2014-12-01

    The degradation of DCF was investigated in aqueous solution by using electron beam (EB) technology. When the initial concentration was between 10 and 40 mg/L, almost 100% of the DCF was degraded at a dose of 0.5 kGy. However, only about 6.5% of DCF was mineralized even at 2 kGy according to total organic carbon (TOC) measurements. A combined process of EB and biological aerated filter (BAF) was therefore developed to enhance the treatment of DCF contaminated surface water. The effluent quality of combined process was substantially improved by EB pretreatment due to the degradation of DCF and related intermediates. Both irradiation and biological treatment reduced the toxicity of the treated water. The experimental results showed that EB is effective for removing DCF from artificial aqueous solution and real surface water.

  2. Fibronectin adsorption, cell adhesion, and proliferation on nanostructured tantalum surfaces.

    Science.gov (United States)

    Dolatshahi-Pirouz, A; Jensen, T; Kraft, David Christian; Foss, Morten; Kingshott, Peter; Hansen, John Lundsgaard; Larsen, Arne Nylandsted; Chevallier, Jacques; Besenbacher, Flemming

    2010-05-25

    The interaction between dental pulp derived mesenchymal stem cells (DP-MSCs) and three different tantalum nanotopographies with and without a fibronectin coating is examined: sputter-coated tantalum surfaces with low surface roughness tantalum surfaces were examined, as well as cellular attachment, proliferation, and vinculin focal adhesion spot assembly on the respective surfaces. The results showed the highest fibronectin mass uptake on the hut structures, with a slightly higher availability of cell-binding domains and the most pronounced formation of vinculin focal adhesion spots as compared to the other surfaces. The proliferation of DP-MSCs was found to be significantly higher on dome and hut surfaces coated with fibronectin compared to the uncoated flat tantalum surfaces. Consequently, the results presented in this study indicate that fibronectin-coated nanotopographies with a vertical dimension of less than 5 nm influence cell adhesion. This rather interesting behavior is argued to originate from the more available fibronectin cell-binding domains observed on the hut structures. PMID:20443575

  3. Manipulating mammalian cell by phase transformed titanium surface fabricated through ultra-short pulsed laser synthesis.

    Science.gov (United States)

    Chinnakkannu Vijayakumar, Sivaprasad; Venkatakrishnan, Krishnan; Tan, Bo

    2016-01-15

    Developing cell sensitive indicators on interacting substrates that allows specific cell manipulation by a combination of physical, chemical or mechanical cues is a challenge for current biomaterials. Hence, various fabrication approaches have been created on a variety of substrates to mimic or create cell specific cues. However, to achieve cell specific cues a multistep process or a post-chemical treatment is often necessitated. So, a simple approach without any chemical or biological treatment would go a long way in developing bio-functionalized substrates to effectively modulate cell adhesion and interaction. The present investigation is aimed to study the manipulative activity induced by phase transformed titanium surface. An ultra-short laser is used to fabricate the phase transformed titanium surface where a polymorphic titanium oxide phases with titanium monoxide (TiO), tri-titanium oxide (Ti3O) and titanium dioxide (TiO2) have been synthesized on commercially pure titanium. Control over oxide phase transformed area was demonstrated via a combination of laser scanning time (laser pulse interaction time) and laser pulse widths (laser pulse to pulse separation time). The interaction of phase transformed titanium surface with NIH3T3 fibroblasts and MC3T3-E1 osteoblast cells developed a new bio-functionalized platforms on titanium based biomaterials to modulate cell migration and adhesion. The synthesized phase transformed titanium surface on the whole appeared to induce directional cues for cell migration with unique preferential cell adhesion unseen by other fabrication approaches. The precise bio-functionalization controllability exhibited during fabrication offers perceptible edge for developing a variety of smart bio-medical devices, implants and cardiovascular stents where the need in supressing specific cell adhesion and proliferation is of great demand.

  4. Estimating intercellular surface tension by laser-induced cell fusion

    International Nuclear Information System (INIS)

    Intercellular surface tension is a key variable in understanding cellular mechanics. However, conventional methods are not well suited for measuring the absolute magnitude of intercellular surface tension because these methods require determination of the effective viscosity of the whole cell, a quantity that is difficult to measure. In this study, we present a novel method for estimating the intercellular surface tension at single-cell resolution. This method exploits the cytoplasmic flow that accompanies laser-induced cell fusion when the pressure difference between cells is large. Because the cytoplasmic viscosity can be measured using well-established technology, this method can be used to estimate the absolute magnitudes of tension. We applied this method to two-cell-stage embryos of the nematode Caenorhabditis elegans and estimated the intercellular surface tension to be in the 30–90 µN m−1 range. Our estimate was in close agreement with cell–medium surface tensions measured at single-cell resolution. (communication)

  5. Combination of surface nanocrystallization and co-rolling: Creating multilayer nanocrystalline composites

    Energy Technology Data Exchange (ETDEWEB)

    Waltz, L. [Institute Charles Delaunay, FRE CNRS 2848, LASMIS, University of Technology of Troyes, 10000 Troyes (France); Retraint, D. [Institute Charles Delaunay, FRE CNRS 2848, LASMIS, University of Technology of Troyes, 10000 Troyes (France)], E-mail: delphine.retraint@utt.fr; Roos, A. [Institute Charles Delaunay, FRE CNRS 2848, LASMIS, University of Technology of Troyes, 10000 Troyes (France); Olier, P. [DEN/DNM/SRMA/LTMEX, Commissariat a l' Energie Atomique de Saclay, 91191 Gif-sur-Yvette (France)

    2009-01-15

    This paper describes a combination of surface mechanical attrition treatment and co-rolling performed at 550 deg. C. This duplex method leads to the formation of a semi-massive multilayer structure of alternating nanocrystalline layers, transition layers and coarse-grained layers. Transmission electron microscopy observations correlated with nanoindentation hardness measurements demonstrate that the nano- and subnanocrystalline layers are preserved after the process. Tensile tests showed improved yield and ultimate strengths, and acceptable ductility.

  6. Wear mechanism of heavy load friction contact pairs in tracked vehicle by combined ferrography and surface analysis

    Institute of Scientific and Technical Information of China (English)

    LI Li; ZHU You-li; HUANG Yuan-lin; XU Bin-shi; LI Xiao-yan

    2004-01-01

    Ferrography is deemed as one of the most effective methods for wear particle analysis and failure diagnosis. By analyzing the configuration, content and composition of wear particles in the lubricanting grease and the surface state of the worn surface with combined ferrography and surface analysis techniques, the wear mechanism of the ball groove of the master clutch's release device of a heavy load tracked vehicle was determined. Results show that the controlling wear mechanism is combined of abrasion, adhesion, contact fatigue and corrosion wear, which demonstrates the effectiveness of using combined ferrography and worn surface analysis for the study of wear mechanism of contact surface with friction.

  7. Cell surface recycling in yeast: mechanisms and machineries.

    Science.gov (United States)

    MacDonald, Chris; Piper, Robert C

    2016-04-15

    Sorting internalized proteins and lipids back to the cell surface controls the supply of molecules throughout the cell and regulates integral membrane protein activity at the surface. One central process in mammalian cells is the transit of cargo from endosomes back to the plasma membrane (PM) directly, along a route that bypasses retrograde movement to the Golgi. Despite recognition of this pathway for decades we are only beginning to understand the machinery controlling this overall process. The budding yeastSaccharomyces cerevisiae, a stalwart genetic system, has been routinely used to identify fundamental proteins and their modes of action in conserved trafficking pathways. However, the study of cell surface recycling from endosomes in yeast is hampered by difficulties that obscure visualization of the pathway. Here we briefly discuss how recycling is likely a more prevalent process in yeast than is widely appreciated and how tools might be built to better study the pathway.

  8. Surface modification of closed plastic bags for adherent cell cultivation

    Science.gov (United States)

    Lachmann, K.; Dohse, A.; Thomas, M.; Pohl, S.; Meyring, W.; Dittmar, K. E. J.; Lindenmeier, W.; Klages, C.-P.

    2011-07-01

    In modern medicine human mesenchymal stem cells are becoming increasingly important. However, a successful cultivation of this type of cells is only possible under very specific conditions. Of great importance, for instance, are the absence of contaminants such as foreign microbiological organisms, i.e., sterility, and the chemical functionalization of the ground on which the cells are grown. As cultivation of these cells makes high demands, a new procedure for cell cultivation has been developed in which closed plastic bags are used. For adherent cell growth chemical functional groups have to be introduced on the inner surface of the plastic bag. This can be achieved by a new, atmospheric-pressure plasma-based method presented in this paper. The method which was developed jointly by the Fraunhofer IST and the Helmholtz HZI can be implemented in automated equipment as is also shown in this contribution. Plasma process gases used include helium or helium-based gas mixtures (He + N2 + H2) and vapors of suitable film-forming agents or precursors such as APTMS, DACH, and TMOS in helium. The effect of plasma treatment is investigated by FTIR-ATR spectroscopy as well as surface tension determination based on contact angle measurements and XPS. Plasma treatment in nominally pure helium increases the surface tension of the polymer foil due to the presence of oxygen traces in the gas and oxygen diffusing through the gas-permeable foil, respectively, reacting with surface radical centers formed during contact with the discharge. Primary amino groups are obtained on the inner surface by treatment in mixtures with nitrogen and hydrogen albeit their amount is comparably small due to diffusion of oxygen through the gas-permeable bag, interfering with the plasma-amination process. Surface modifications introducing amino groups on the inner surface turned out to be most efficient in the promotion of cell growth.

  9. Effect of hydroxyapatite surface morphology on cell adhesion.

    Science.gov (United States)

    Iwamoto, Takashi; Hieda, Yohki; Kogai, Yasumichi

    2016-12-01

    We obtained hydroxyapatite (HAp) materials as a block by mixing HAp nanoparticles and polymer, and then calcining the mixtures. The surface morphology of the HAp materials was tuned by varying heat treatment conditions. After calcining the mixtures at 1200 or 800°C for 4h, the surface morphology of the HAp materials was flat or convexo-concave, respectively. The flat surface morphology, which showed micrometer-ordered grain boundaries, was formed by the aggregation of HAp nanoparticles. On the other hand, the convexo-concave surface morphology resulted from the agglomeration of HAp nanoparticles after heat treatment at 800°C for 4h with nanometer-ordered particle size. We tested cell adhesion to HAp materials with flat or convexo-concave surface morphology and found that cells adhered well to the flat HAp materials but not to the convexo-concave HAp materials. This technique for selectively preparing HAp materials with flat or convexo-concave surface morphology was very easy because we merely mixed commercial HAp nanoparticles with polymer and then calcined the mixtures. As a result, the heat treatment temperature affected the surface morphology of our HAp materials, and their surface morphologies contributed to cell adhesion independently of other material properties. PMID:27612825

  10. Accelerated cell-surface interlocking on plasma polymer-modified porous ceramics.

    Science.gov (United States)

    Rebl, Henrike; Finke, Birgit; Schmidt, Jürgen; Mohamad, Heba S; Ihrke, Roland; Helm, Christiane A; Nebe, J Barbara

    2016-12-01

    Excellent osseointegration of permanent implants is crucial for the long lasting success of the implantation. To improve the osseointegrative potential, bio-inert titanium alloy surfaces (Ti6Al4V) are modified by plasma chemical oxidation (PCO®) of the titanium-oxide layer to a non-stoichiometric, amorphous calcium phosphate layer. The native titanium-oxide film measuring only a few nanometers is converted by PCO® to a thick porous calcium phosphate layer of about 10μm. In a second step the PCO surface is combined with a cell adhesive plasma-polymerized allylamine (PPAAm) nano film (5 and 50nm). Independent of the PPAAm coating homogeneity, the human osteoblast-like MG-63 cells show a remarkable increase in cell size and well-developed filopodia. Analyses of the actin cytoskeleton reveal that the cells mold to the pore shape of the PPAAm-covered PCO, thereby establishing a strong attachment to the surface. Interestingly, we could demonstrate that even though our untreated PCO shows excellent hydrophilicity, this alone is not sufficient to facilitate fast cell spreading, but the positive surface charges mediated by PPAAm. This multilayer composite material guarantees enhanced interlocking of the cells with the porous surface. PMID:27612809

  11. A cell cycle and nutritional checkpoint controlling bacterial surface adhesion.

    Directory of Open Access Journals (Sweden)

    Aretha Fiebig

    2014-01-01

    Full Text Available In natural environments, bacteria often adhere to surfaces where they form complex multicellular communities. Surface adherence is determined by the biochemical composition of the cell envelope. We describe a novel regulatory mechanism by which the bacterium, Caulobacter crescentus, integrates cell cycle and nutritional signals to control development of an adhesive envelope structure known as the holdfast. Specifically, we have discovered a 68-residue protein inhibitor of holdfast development (HfiA that directly targets a conserved glycolipid glycosyltransferase required for holdfast production (HfsJ. Multiple cell cycle regulators associate with the hfiA and hfsJ promoters and control their expression, temporally constraining holdfast development to the late stages of G1. HfiA further functions as part of a 'nutritional override' system that decouples holdfast development from the cell cycle in response to nutritional cues. This control mechanism can limit surface adhesion in nutritionally sub-optimal environments without affecting cell cycle progression. We conclude that post-translational regulation of cell envelope enzymes by small proteins like HfiA may provide a general means to modulate the surface properties of bacterial cells.

  12. Ionized dopant concentrations at the heavily doped surface of a silicon solar cell

    Science.gov (United States)

    Weinberg, I.; Broder, J. D.; Mazaris, G. A., Jr.; Hsu, L.

    1978-01-01

    Data are combined with concentrations obtained by a bulk measurement method using successive layer removal with measurements of Hall effect and resistivity. From the MOS (metal-oxide-semiconductor) measurements it is found that the ionized dopant concentration N has the value (1.4 + or - 0.1) x 10 to the 20th power/cu cm at distances between 100 and 220 nm from the n(+) surface. The bulk measurement technique yields average values of N over layers whose thickness is 2000 nm. Results show that, at the higher concentrations encountered at the n(+) surface, the MOS C-V technique, when combined with a bulk measurement method, can be used to evaluate the effects of materials preparation methodologies on the surface and near surface concentrations of silicon cells.

  13. Combining Spectral and Texture Features Using Random Forest Algorithm: Extracting Impervious Surface Area in Wuhan

    Science.gov (United States)

    Shao, Zhenfeng; Zhang, Yuan; Zhang, Lei; Song, Yang; Peng, Minjun

    2016-06-01

    Impervious surface area (ISA) is one of the most important indicators of urban environments. At present, based on multi-resolution remote sensing images, numerous approaches have been proposed to extract impervious surface, using statistical estimation, sub-pixel classification and spectral mixture analysis method of sub-pixel analysis. Through these methods, impervious surfaces can be effectively applied to regional-scale planning and management. However, for the large scale region, high resolution remote sensing images can provide more details, and therefore they will be more conducive to analysis environmental monitoring and urban management. Since the purpose of this study is to map impervious surfaces more effectively, three classification algorithms (random forests, decision trees, and artificial neural networks) were tested for their ability to map impervious surface. Random forests outperformed the decision trees, and artificial neural networks in precision. Combining the spectral indices and texture, random forests is applied to impervious surface extraction with a producer's accuracy of 0.98, a user's accuracy of 0.97, and an overall accuracy of 0.98 and a kappa coefficient of 0.97.

  14. Cell surface differences of Naegleria fowleri and Naegleria lovaniensis exposed with surface markers.

    Science.gov (United States)

    González-Robles, Arturo; Castañón, Guadalupe; Cristóbal-Ramos, Ana Ruth; Hernández-Ramírez, Verónica Ivonne; Omaña-Molina, Maritza; Martínez-Palomo, Adolfo

    2007-12-01

    Differences in the distribution of diverse cell surface coat markers were found between Naegleria fowleri and Naegleria lovaniensis. The presence of carbohydrate-containing components in the cell coat of the two species was detected by selective staining with ruthenium red and alcian blue. Using both markers, N. fowleri presented a thicker deposit than N. lovaniensis. The existence of exposed mannose or glucose residues was revealed by discriminatory agglutination with the plant lectin Concanavalin A. These sugar residues were also visualized at the cell surface of these parasites either by transmission electron microscopy or by fluorescein-tagged Concanavalin A. Using this lectin cap formation was induced only in N. fowleri. The anionic sites on the cell surface detected by means of cationized ferritin were more apparent in N. fowleri. Biotinylation assays confirmed that even though the two amoebae species have some analogous plasma membrane proteins, there is a clear difference in their composition.

  15. Effective combination of aligned nanocomposite nanofibers and human unrestricted somatic stem cells for bone tissue engineering

    Institute of Scientific and Technical Information of China (English)

    Behnaz BAKHSHANDEH; Masoud SOLEIMANI; Nasser GHAEMI; Iman SHABANI

    2011-01-01

    Aim: Bioartificial bone tissue engineering is an increasingly popular technique to solve bone defect challenges. This study aimed to investigate the interactions between matrix composition and appropriate cell type, focusing on hydroxyapatite (HA), to achieve a more effective combination for bone regeneration.Methods: Human unrestricted somatic stem cells (USSCs) were isolated from placental cord blood. The cellular and molecular events during the osteo-induction of USSCs were evaluated for 21 d under the following conditions: (1) in basal culture, (2) supplemented with hydroxyapatite nanoparticle (nHA) suspension, and (3) seeded on electrospun aligned nanoflbrous poly-ε-caprolactone/poly-L-lactic acid/nHA (PCL/PLLA/nHA) scaffolds. The scaffolds were characterized using scanning electron microscope (SEM), fourier transform infrared spectroscopy (FTIR) and tensile test.Results: Maintenance of USSCs for 21 d in basal or osteogenic culture resulted in significant increase in osteoblast differentiation. With nHA suspension, even soluble osteo-inductive additives were ineffective, probably due to induced apoptosis of the cells. In con-trast to the hindrance of proliferation by nHA suspension, the scaffolds improved cell growth. The scaffolds mimic the nanostructure of natural bone matrix with the combination of PLLA/PCL (organic phase) and HA (inorganic phase) offering a favorable surface topogra-phy, which was demonstrated to possess suitable properties for supporting USSCs. Quantitative measurement of osteogenic markers, enzymatic activity and mineralization indicated that the scaffolds did not disturb, but enhanced the osteogenic potential of USSCs.Moreover, the alignment of the fibers led to cell orientation during cell growth.Conclusion: The results demonstrated the synergism of PCL/PLLA/nHA nanoflbrous scaffolds and USSCs in the augmentation of osteo-genic differentiation. Thus, nHA grafted into PCI./PLLA scaffolds can be a suitable choice for bone tissue

  16. Estimating Sea Surface Salinity and Wind Using Combined Passive and Active L-Band Microwave Observations

    Science.gov (United States)

    Yueh, Simon H.; Chaubell, Mario J.

    2012-01-01

    Several L-band microwave radiometer and radar missions have been, or will be, operating in space for land and ocean observations. These include the NASA Aquarius mission and the Soil Moisture Active Passive (SMAP) mission, both of which use combined passive/ active L-band instruments. Aquarius s passive/active L-band microwave sensor has been designed to map the salinity field at the surface of the ocean from space. SMAP s primary objectives are for soil moisture and freeze/thaw detection, but it will operate continuously over the ocean, and hence will have significant potential for ocean surface research. In this innovation, an algorithm has been developed to retrieve simultaneously ocean surface salinity and wind from combined passive/active L-band microwave observations of sea surfaces. The algorithm takes advantage of the differing response of brightness temperatures and radar backscatter to salinity, wind speed, and direction, thus minimizing the least squares error (LSE) measure, which signifies the difference between measurements and model functions of brightness temperatures and radar backscatter. The algorithm uses the conjugate gradient method to search for the local minima of the LSE. Three LSE measures with different measurement combinations have been tested. The first LSE measure uses passive microwave data only with retrieval errors reaching 1 to 2 psu (practical salinity units) for salinity, and 1 to 2 m/s for wind speed. The second LSE measure uses both passive and active microwave data for vertical and horizontal polarizations. The addition of active microwave data significantly improves the retrieval accuracy by about a factor of five. To mitigate the impact of Faraday rotation on satellite observations, the third LSE measure uses measurement combinations invariant under the Faraday rotation. For Aquarius, the expected RMS SSS (sea surface salinity) error will be less than about 0.2 psu for low winds, and increases to 0.3 psu at 25 m/s wind speed

  17. Reversed cell imprinting, AFM imaging and adhesion analyses of cells on patterned surfaces.

    Science.gov (United States)

    Zhou, Xiongtu; Shi, Jian; Zhang, Fan; Hu, Jie; Li, Xin; Wang, Li; Ma, Xueming; Chen, Yong

    2010-05-01

    Cell adhesion and motility depend strongly on the interactions between cells and cell culture substratum. To observe the cell morphology at the interface between cells and artificial substratum or patterned surfaces, we have developed a technique named reversed cell imprinting. After culture and chemical fixation of the cells on a patterned hole array, a liquid polymer was poured on and UV cured, allowing taking off the cell-polymer assembly for a direct observation of the underside cell surface using atomic force microscopy. As expected, we observed local deformation of the cell membrane in the hole area with a penetration depth strongly dependent on the size and depth of the hole as well as the culture time. Quantitative analyses of Hela cells on patterned surfaces of polydimethylsiloxane (PDMS) revealed that the penetration was also position dependent over the cell attachment area due to the non-homogeneous distribution of the membrane stress. With the increase of the culture time, the penetration depth was reduced, in a close correlation with the increase of the cell spreading area. Nevertheless, both cell seeding and adhesion efficiency on high density hole arrays could be significantly increased comparing to that on a smooth surface. Patterned substrates are increasingly required to produce and interrogate new biomaterials for therapeutic benefit. Overall, this work suggests a strategy to endow conventional imaging methods with added functionality to enable easy observation of the underside cell morphology on topographic patterns. PMID:20390138

  18. Revealing Surface Waters on an Antifreeze Protein by Fusion Protein Crystallography Combined with Molecular Dynamic Simulations.

    Science.gov (United States)

    Sun, Tianjun; Gauthier, Sherry Y; Campbell, Robert L; Davies, Peter L

    2015-10-01

    Antifreeze proteins (AFPs) adsorb to ice through an extensive, flat, relatively hydrophobic surface. It has been suggested that this ice-binding site (IBS) organizes surface waters into an ice-like clathrate arrangement that matches and fuses to the quasi-liquid layer on the ice surface. On cooling, these waters join the ice lattice and freeze the AFP to its ligand. Evidence for the generality of this binding mechanism is limited because AFPs tend to crystallize with their IBS as a preferred protein-protein contact surface, which displaces some bound waters. Type III AFP is a 7 kDa globular protein with an IBS made up two adjacent surfaces. In the crystal structure of the most active isoform (QAE1), the part of the IBS that docks to the primary prism plane of ice is partially exposed to solvent and has clathrate waters present that match this plane of ice. The adjacent IBS, which matches the pyramidal plane of ice, is involved in protein-protein crystal contacts with few surface waters. Here we have changed the protein-protein contacts in the ice-binding region by crystallizing a fusion of QAE1 to maltose-binding protein. In this 1.9 Å structure, the IBS that fits the pyramidal plane of ice is exposed to solvent. By combining crystallography data with MD simulations, the surface waters on both sides of the IBS were revealed and match well with the target ice planes. The waters on the pyramidal plane IBS were loosely constrained, which might explain why other isoforms of type III AFP that lack the prism plane IBS are less active than QAE1. The AFP fusion crystallization method can potentially be used to force the exposure to solvent of the IBS on other AFPs to reveal the locations of key surface waters.

  19. Revealing Surface Waters on an Antifreeze Protein by Fusion Protein Crystallography Combined with Molecular Dynamic Simulations.

    Science.gov (United States)

    Sun, Tianjun; Gauthier, Sherry Y; Campbell, Robert L; Davies, Peter L

    2015-10-01

    Antifreeze proteins (AFPs) adsorb to ice through an extensive, flat, relatively hydrophobic surface. It has been suggested that this ice-binding site (IBS) organizes surface waters into an ice-like clathrate arrangement that matches and fuses to the quasi-liquid layer on the ice surface. On cooling, these waters join the ice lattice and freeze the AFP to its ligand. Evidence for the generality of this binding mechanism is limited because AFPs tend to crystallize with their IBS as a preferred protein-protein contact surface, which displaces some bound waters. Type III AFP is a 7 kDa globular protein with an IBS made up two adjacent surfaces. In the crystal structure of the most active isoform (QAE1), the part of the IBS that docks to the primary prism plane of ice is partially exposed to solvent and has clathrate waters present that match this plane of ice. The adjacent IBS, which matches the pyramidal plane of ice, is involved in protein-protein crystal contacts with few surface waters. Here we have changed the protein-protein contacts in the ice-binding region by crystallizing a fusion of QAE1 to maltose-binding protein. In this 1.9 Å structure, the IBS that fits the pyramidal plane of ice is exposed to solvent. By combining crystallography data with MD simulations, the surface waters on both sides of the IBS were revealed and match well with the target ice planes. The waters on the pyramidal plane IBS were loosely constrained, which might explain why other isoforms of type III AFP that lack the prism plane IBS are less active than QAE1. The AFP fusion crystallization method can potentially be used to force the exposure to solvent of the IBS on other AFPs to reveal the locations of key surface waters. PMID:26371748

  20. Combined cisplatin and aurora inhibitor treatment increase neuroblastoma cell death but surviving cells overproduce BDNF.

    Science.gov (United States)

    Polacchini, Alessio; Albani, Clara; Baj, Gabriele; Colliva, Andrea; Carpinelli, Patrizia; Tongiorgi, Enrico

    2016-07-15

    Drug-resistance to chemotherapics in aggressive neuroblastoma (NB) is characterized by enhanced cell survival mediated by TrkB and its ligand, brain-derived neurotrophic factor (BDNF); thus reduction in BDNF levels represent a promising strategy to overcome drug-resistance, but how chemotherapics regulate BDNF is unknown. Here, cisplatin treatment in SK-N-BE neuroblastoma upregulated multiple BDNF transcripts, except exons 5 and 8 variants. Cisplatin increased BDNF mRNA and protein, and enhanced translation of a firefly reporter gene flanked by BDNF 5'UTR exons 1, 2c, 4 or 6 and 3'UTR-long. To block BDNF translation we focused on aurora kinases inhibitors which are proposed as new chemotherapeutics. NB cell survival after 24 h treatment was 43% with cisplatin, and 22% by cisplatin+aurora kinase inhibitor PHA-680632, while the aurora kinases inhibitor alone was less effective; however the combined treatment induced a paradoxical increase of BDNF in surviving cells with strong translational activation of exon6-3'UTR-long transcript, while translation of BDNF transcripts 1, 2C and 4 was suppressed. In conclusion, combined cisplatin and aurora kinase inhibitor treatment increases cell death, but induces BDNF overproduction in surviving cells through an aurora kinase-independent mechanism.

  1. Hepatic Bel-7402 Cell Proliferation on Different Phospholipid Surfaces

    Institute of Scientific and Technical Information of China (English)

    1999-01-01

    Phospholipids are believed to be important biomaterials.However, limited information is available on their cytocompatibilities.The objective of this study is to evaluate the effects of different phospholipids on the proliferation of hepatic Bel-7402 cells by comparing the adhesion, viability and proliferation of Bel-7402 cells cultured on different phospholipid surfaces.The cell adhesion, determined by counting the number of adhered cells to the surface, indicated that the cell adhesion was enhanced on charged phospolipid membranes.The cell viability evaluated by MTT[3 (4,5-dimethylthiazole-2-yl)-2,5-diphenyl tetrazolium-bromide] showed that cells cultured on charged phospholipids have greater viability than those cultured on the control, while cells cultured on neutral phospholipids showed lower viability.The cell cycle analysis using flow cytometry demonstrated that S phase entry increased on charged phospholipids, while S phase entry decreased on neutral phospholipids.The results suggested that charged phospholipids, especially positively charged phospholipids, show better cytocompatibilities than neutral phospholipids to hepatic Bel-7402 cell.

  2. Origin of subdiffusion of water molecules on cell membrane surfaces

    CERN Document Server

    Yamamoto, Eiji; Yasui, Masato; Yasuoka, Kenji

    2014-01-01

    Water molecules play an important role in providing unique environments for biological reactions on cell membranes. It is widely believed that water molecules form bridges that connect lipid molecules and stabilize cell membranes. Using all-atom molecular dynamics simulations, we show that translational and rotational diffusion of water molecules on lipid membrane surfaces exhibit subdiffusion. Moreover, we provide evidence that both divergent mean trapping time (continuous-time random walk) and long-correlated noise (fractional Brownian motion) contribute to this subdiffusion. These results suggest that subdiffusion on cell membranes causes the water retardation, an enhancement of cell membrane stability, and a higher reaction efficiency.

  3. Mild hypothermia combined with neural stem cell transplantation for hypoxic-ischemic encephalopathy:neuroprotective effects of combined therapy

    Institute of Scientific and Technical Information of China (English)

    Lin Wang; Feng Jiang; Qifeng Li; Xiaoguang He; Jie Ma

    2014-01-01

    Neural stem cell transplantation is a useful treatment for ischemic stroke, but apoptosis often occurs in the hypoxic-ischemic environment of the brain after cell transplantation. In this study, we determined if mild hypothermia (27-28°C) can increase the survival rate of neural stem cells (1.0 × 105 /μL) transplanted into neonatal mice with hypoxic-ischemic encephalopathy. Long-term effects on neurological functioning of the mice were also examined. After mild hy-pothermia combined with neural stem cell transplantation, we observed decreased expression levels of inflammatory factor nuclear factor-kappa B and apoptotic factor caspase-3, reduced cerebral infarct volumes, increased survival rate of transplanted cells, and marked improvements in neurological function. Thus, the neuroprotective effects of mild hypothermia combined with neural stem cell transplantation are superior to those of monotherapy. Moreover, our ifndings suggest that the neuroprotective effects of mild hypothermia combined with neural stem cell transplantation on hypoxic-ischemic encephalopathy are achieved by anti-inlfammatory and an-ti-apoptotic mechanisms.

  4. Mild hypothermia combined with neural stem cell transplantation for hypoxic-ischemic encephalopathy: neuroprotective effects of combined therapy.

    Science.gov (United States)

    Wang, Lin; Jiang, Feng; Li, Qifeng; He, Xiaoguang; Ma, Jie

    2014-10-01

    Neural stem cell transplantation is a useful treatment for ischemic stroke, but apoptosis often occurs in the hypoxic-ischemic environment of the brain after cell transplantation. In this study, we determined if mild hypothermia (27-28°C) can increase the survival rate of neural stem cells (1.0 × 10(5)/μL) transplanted into neonatal mice with hypoxic-ischemic encephalopathy. Long-term effects on neurological functioning of the mice were also examined. After mild hypothermia combined with neural stem cell transplantation, we observed decreased expression levels of inflammatory factor nuclear factor-kappa B and apoptotic factor caspase-3, reduced cerebral infarct volumes, increased survival rate of transplanted cells, and marked improvements in neurological function. Thus, the neuroprotective effects of mild hypothermia combined with neural stem cell transplantation are superior to those of monotherapy. Moreover, our findings suggest that the neuroprotective effects of mild hypothermia combined with neural stem cell transplantation on hypoxic-ischemic encephalopathy are achieved by anti-inflammatory and anti-apoptotic mechanisms. PMID:25422635

  5. A combined QCM and XPS investigation of asphaltene adsorption on metal surfaces.

    Science.gov (United States)

    Rudrake, Amit; Karan, Kunal; Horton, J Hugh

    2009-04-01

    To investigate asphaltene-metal interactions, a combined quartz crystal microbalance (QCM) and X-ray photoelectron spectroscopy (XPS) study of asphaltene adsorption on a gold surface was conducted. Adsorption experiments were conducted at 25 degrees C with solutions of asphaltenes in toluene at concentrations ranging from 50 to 1500 ppm. QCM measurements yielded information on the kinetics of adsorption and further assessment of the data allowed the estimation of equilibrium adsorption levels. XPS analysis of adsorbed and bulk asphaltene demonstrated the presence of carboxylic, thiophenic, sulfide, pyridinic and pyrrolic type functional groups. The intensity of the main carbon (C-H) peak was related to surface coverage of adsorbed asphaltene as a function of asphaltene concentration by a simple mathematical model. The mass adsorption data from the QCM experiments also allowed estimation of the surface coverage, which was compared to those from XPS analyses. Surface coverage estimates as a function of asphaltene concentration could be described by a Langmuir (type-I) isotherm. The free energy of asphaltene adsorption was estimated to be -26.8+/-0.1 and -27.3+/-0.1 kJ/mol from QCM and XPS data, respectively assuming asphaltene molar mass of 750 g/gmol. QCM and XPS data was also analyzed to estimate adsorbed layer thickness after accounting for surface coverage. The thickness of the adsorbed asphaltene estimated from both XPS and QCM data analyses ranged from 6-8 nm over the entire range of adsorption concentrations investigated.

  6. Flotation separation of polyvinyl chloride and polyethylene terephthalate plastics combined with surface modification for recycling.

    Science.gov (United States)

    Wang, Chongqing; Wang, Hui; Fu, Jiangang; Zhang, Lingling; Luo, Chengcheng; Liu, Younian

    2015-11-01

    Surface modification with potassium permanganate (KMnO4) solution was developed for separation of polyvinyl chloride (PVC) and polyethylene terephthalate (PET) waste plastics. The floatability of PVC decreases with increasing of KMnO4 concentration, treatment time, temperature and stirring rate, while that of PET is unaffected. Fourier transform infrared (FT-IR) analysis confirms that mechanism of surface modification may be due to oxidization reactions occurred on PVC surface. The optimum conditions are KMnO4 concentration 1.25 mM/L, treatment time 50 min, temperature 60°C, stirring rate 300 r/min, frother concentration 17.5 g/L and flotation time 1 min. PVC and PET with different particle sizes were separated efficiently through two-stage flotation. Additionally, after ultrasonic assisted surface modification, separation of PVC and PET with different mass ratios was obtained efficiently through one-stage flotation. The purity and the recovery of the obtained products after flotation separation are up to 99.30% and 99.73%, respectively. A flotation process was designed for flotation separation of PVC and PET plastics combined with surface modification. This study provides technical insights into physical separation of plastic wastes for recycling industry. PMID:26253330

  7. Flotation separation of polyvinyl chloride and polyethylene terephthalate plastics combined with surface modification for recycling.

    Science.gov (United States)

    Wang, Chongqing; Wang, Hui; Fu, Jiangang; Zhang, Lingling; Luo, Chengcheng; Liu, Younian

    2015-11-01

    Surface modification with potassium permanganate (KMnO4) solution was developed for separation of polyvinyl chloride (PVC) and polyethylene terephthalate (PET) waste plastics. The floatability of PVC decreases with increasing of KMnO4 concentration, treatment time, temperature and stirring rate, while that of PET is unaffected. Fourier transform infrared (FT-IR) analysis confirms that mechanism of surface modification may be due to oxidization reactions occurred on PVC surface. The optimum conditions are KMnO4 concentration 1.25 mM/L, treatment time 50 min, temperature 60°C, stirring rate 300 r/min, frother concentration 17.5 g/L and flotation time 1 min. PVC and PET with different particle sizes were separated efficiently through two-stage flotation. Additionally, after ultrasonic assisted surface modification, separation of PVC and PET with different mass ratios was obtained efficiently through one-stage flotation. The purity and the recovery of the obtained products after flotation separation are up to 99.30% and 99.73%, respectively. A flotation process was designed for flotation separation of PVC and PET plastics combined with surface modification. This study provides technical insights into physical separation of plastic wastes for recycling industry.

  8. Bottom-up engineering of the surface roughness of nanostructured cubic zirconia to control cell adhesion.

    Science.gov (United States)

    Singh, A V; Ferri, M; Tamplenizza, M; Borghi, F; Divitini, G; Ducati, C; Lenardi, C; Piazzoni, C; Merlini, M; Podestà, A; Milani, P

    2012-11-30

    Nanostructured cubic zirconia is a strategic material for biomedical applications since it combines superior structural and optical properties with a nanoscale morphology able to control cell adhesion and proliferation. We produced nanostructured cubic zirconia thin films at room temperature by supersonic cluster beam deposition of nanoparticles produced in the gas phase. Precise control of film roughness at the nanoscale is obtained by operating in a ballistic deposition regime. This allows one to study the influence of nanoroughness on cell adhesion, while keeping the surface chemistry constant. We evaluated cell adhesion on nanostructured zirconia with an osteoblast-like cell line using confocal laser scanning microscopy for detailed morphological and cytoskeleton studies. We demonstrated that the organization of cytoskeleton and focal adhesion formation can be controlled by varying the evolution of surface nanoroughness.

  9. Investigation of the Cell Surface Proteome of Human Periodontal Ligament Stem Cells.

    Science.gov (United States)

    Xiong, Jimin; Menicanin, Danijela; Zilm, Peter S; Marino, Victor; Bartold, P Mark; Gronthos, Stan

    2016-01-01

    The present study examined the cell surface proteome of human periodontal ligament stem cells (PDLSC) compared to human fibroblasts. Cell surface proteins were prelabelled with CyDye before processing to extract the membrane lysates, which were separated using 2D electrophoresis. Selected differentially expressed protein "spots" were identified using Mass spectrometry. Four proteins were selected for validation: CD73, CD90, Annexin A2, and sphingosine kinase 1 previously associated with mesenchymal stem cells. Flow cytometric analysis found that CD73 and CD90 were highly expressed by human PDLSC and gingival fibroblasts but not by keratinocytes, indicating that these antigens could be used as potential markers for distinguishing between mesenchymal cells and epithelial cell populations. Annexin A2 was also found to be expressed at low copy number on the cell surface of human PDLSC and gingival fibroblasts, while human keratinocytes lacked any cell surface expression of Annexin A2. In contrast, sphingosine kinase 1 expression was detected in all the cell types examined using immunocytochemical analysis. These proteomic studies form the foundation to further define the cell surface protein expression profile of PDLSC in order to better characterise this cell population and help develop novel strategies for the purification of this stem cell population. PMID:27579043

  10. Adhesion defective BHK cell mutant has cell surface heparan sulfate proteoglycan of altered properties

    DEFF Research Database (Denmark)

    Couchman, J R; Austria, R; Woods, A;

    1988-01-01

    sulfation, reduced affinity for fibronectin and decreased half-life on the cell surface when compared to the normal counterpart. Our conclusions based on this data are that these altered properties may, in part, account for the adhesion defect in the ricin-resistant mutant. Whether this results from......In the light of accumulating data that implicate cell surface heparan sulfate proteoglycans (HSPGs) with a role in cell interactions with extracellular matrix molecules such as fibronectin, we have compared the properties of these molecules in wild-type BHK cells and an adhesion-defective ricin......-resistant mutant (RicR14). Our results showed that the mutant, unlike BHK cells, cannot form focal adhesions when adherent to planar substrates in the presence of serum. Furthermore, while both cell lines possess similar amounts of cell surface HSPG with hydrophobic properties, that of RicR14 cells had decreased...

  11. Generating daily high spatial land surface temperatures by combining ASTER and MODIS land surface temperature products for environmental process monitoring.

    Science.gov (United States)

    Wu, Mingquan; Li, Hua; Huang, Wenjiang; Niu, Zheng; Wang, Changyao

    2015-08-01

    There is a shortage of daily high spatial land surface temperature (LST) data for use in high spatial and temporal resolution environmental process monitoring. To address this shortage, this work used the Spatial and Temporal Adaptive Reflectance Fusion Model (STARFM), Enhanced Spatial and Temporal Adaptive Reflectance Fusion Model (ESTARFM), and the Spatial and Temporal Data Fusion Approach (STDFA) to estimate high spatial and temporal resolution LST by combining Advanced Spaceborne Thermal Emission and Reflection Radiometer (ASTER) LST and Moderate Resolution Imaging Spectroradiometer (MODIS) LST products. The actual ASTER LST products were used to evaluate the precision of the combined LST images using the correlation analysis method. This method was tested and validated in study areas located in Gansu Province, China. The results show that all the models can generate daily synthetic LST image with a high correlation coefficient (r) of 0.92 between the synthetic image and the actual ASTER LST observations. The ESTARFM has the best performance, followed by the STDFA and the STARFM. Those models had better performance in desert areas than in cropland. The STDFA had better noise immunity than the other two models. PMID:26165141

  12. Enhanced cell attachment using a novel cell culture surface presenting functional domains from extracellular matrix proteins

    OpenAIRE

    Cooke, M. J.; Phillips, S R; Shah, D. S. H.; Athey, D.; Lakey, J H; Przyborski, S A

    2008-01-01

    Many factors contribute to the creation and maintenance of a realistic environment for cell growth in vitro, e.g. the consistency of the growth medium, the addition of supplements, and the surface on which the cells grow. The nature of the surface on which cells are cultured plays an important role in their ability to attach, proliferate, migrate and function. Components of the extracellular matrix (ECM) are often used to coat glass or plastic surfaces to enhance cell attachment in vitro. Fra...

  13. Combined experiment and theory approach in surface chemistry: Stairway to heaven?

    Science.gov (United States)

    Exner, Kai S.; Heß, Franziska; Over, Herbert; Seitsonen, Ari Paavo

    2015-10-01

    In this perspective we discuss how an intimate interaction of experiments with theory is able to deepen our insight into the catalytic reaction system on the molecular level. This strategy is illustrated by discussing various examples from our own research of surface chemistry and model catalysis. The particular examples were carefully chosen to balance the specific strength of both approaches - theory and experiment - and emphasize the benefit of this combined approach. We start with the determination of complex surface structures, where diffraction techniques in combination with theory are clear-cut. The promoter action of alkali metals in heterogeneous catalysis is rationalized with theory and experiment for the case of CO coadsorption. Predictive power of theory is limited as demonstrated with the apparent activity of chlorinated TiO2(110) in the oxidation of HCl: Even if we know all elementary reaction steps of a catalytic reaction mechanism, the overall kinetics may remain elusive and require the application kinetic Monte Carlo simulations. Catalysts are not always stable under reaction conditions and may chemically transform as discussed for the CO oxidation reaction over ruthenium. Under oxidizing reaction conditions ruthenium transforms into RuO2, a process which is hardly understood on the molecular level. Lastly we focus on electrochemical reactions. Here theory is clearly ahead since spectroscopic methods are not available to resolve the processes at the electrode surface.

  14. Immunogold labels: cell-surface markers in atomic force microscopy

    OpenAIRE

    Putman, Constant A.J.; Grooth, de, B.G.; Hansma, Paul K.; Hulst, van der, R.W.M.; Greve, Jan

    1993-01-01

    The feasibility of using immunogold labels as cell-surface markers in atomic force microscopy is shown in this paper. The atomic force microscope (AFM) was used to image the surface of immunogold-labeled human lymphocytes. The lymphocytes were isolated from whole blood and labeled by an indirect immunolabeling method using the monoclonal antibody anti-CD3 and a secondary antibody (Goat-anti-Mouse) linked to 30 nm colloidal gold particles. Some of the samples were enhanced by silver deposition...

  15. Cell surface polypeptides of murine T-cell clones expressing cytolytic or amplifier activity.

    OpenAIRE

    Sarmiento, M.; Glasebrook, A L; Fitch, F. W.

    1980-01-01

    Murine cytolytic T-cell and amplifier T-cell clones derived from secondary unidirectional mixed leukocyte cultures were labeled with 125I by the lactoperoxidase method and their polypeptide profiles were analyzed by NaDodSO4/polyacrylamide gel electrophoresis. All cytolytic T-cell clones derived from the same mouse strain yeilded similar cell surface polypeptide profiles. However, profiles obtained with three amplifier T-cell clones were strikingly different from each other as well as from th...

  16. 3D surface topology guides stem cell adhesion and differentiation.

    Science.gov (United States)

    Viswanathan, Priyalakshmi; Ondeck, Matthew G; Chirasatitsin, Somyot; Ngamkham, Kamolchanok; Reilly, Gwendolen C; Engler, Adam J; Battaglia, Giuseppe

    2015-06-01

    Polymerized high internal phase emulsion (polyHIPE) foams are extremely versatile materials for investigating cell-substrate interactions in vitro. Foam morphologies can be controlled by polymerization conditions to result in either open or closed pore structures with different levels of connectivity, consequently enabling the comparison between 2D and 3D matrices using the same substrate with identical surface chemistry conditions. Additionally, here we achieve the control of pore surface topology (i.e. how different ligands are clustered together) using amphiphilic block copolymers as emulsion stabilizers. We demonstrate that adhesion of human mesenchymal progenitor (hES-MP) cells cultured on polyHIPE foams is dependent on foam surface topology and chemistry but is independent of porosity and interconnectivity. We also demonstrate that the interconnectivity, architecture and surface topology of the foams has an effect on the osteogenic differentiation potential of hES-MP cells. Together these data demonstrate that the adhesive heterogeneity of a 3D scaffold could regulate not only mesenchymal stem cell attachment but also cell behavior in the absence of soluble growth factors.

  17. The Role of Surface Receptor Density in Surface-Initiated Polymerizations for Cancer Cell Isolation.

    Science.gov (United States)

    Lilly, Jacob L; Berron, Brad J

    2016-06-01

    Fluid biopsies potentially offer a minimally invasive alternative to traditional tissue biopsies for the continual monitoring of metastatic cancer. Current established technologies for isolating circulating tumor cells (CTCs) suffer from poor purity and yield and require fixatives that preclude the collection of viable cells for longitudinal analyses of biological function. Antigen specific lysis (ASL) is a rapid, high-purity method of cell isolation based on targeted protective coatings on antigen-presenting cells and lysis depletion of unprotected antigen-negative cells. In ASL, photoinitiators are specifically labeled on cell surfaces that enable subsequent surface-initiated polymerization. Critically, the significant determinants of process yield have yet to be investigated for this emerging technology. In this work, we show that the labeling density of photoinitiators is strongly correlated with the yield of intact cells during ASL by flow cytometry analysis. Results suggest ASL is capable of delivering ∼25% of targeted cells after isolation using traditional antibody labeling approaches. Monomer formulations of two molecular weights of PEG-diacrylate (Mn ∼ 575 and 3500) are examined. The gelation response during ASL polymerization is also investigated via protein microarray analogues on planar glass. Finally, a density threshold of photoinitiator labeling required for protection during lysis is determined for both monomer formulations. These results indicate ASL is a promising technology for high yield CTC isolation for rare-cell function assays and fluid biopsies. PMID:27206735

  18. Tetraploid cells from cytokinesis failure induce aneuploidy and spontaneous transformation of mouse ovarian surface epithelial cells

    OpenAIRE

    Lv, Lei; Zhang, Tianwei; Yi, Qiyi; Huang, Yun; Wang, Zheng; Hou, Heli; Zhang, Huan; Zheng, Wei; Hao, Qiaomei; Guo, Zongyou; Howard J Cooke; Shi, Qinghua

    2012-01-01

    Most ovarian cancers originate from the ovarian surface epithelium and are characterized by aneuploid karyotypes. Aneuploidy, a consequence of chromosome instability, is an early event during the development of ovarian cancers. However, how aneuploid cells are evolved from normal diploid cells in ovarian cancers remains unknown. In the present study, cytogenetic analyses of a mouse syngeneic ovarian cancer model revealed that diploid mouse ovarian surface epithelial cells (MOSECs) experienced...

  19. Monoclonal antibody to human endothelial cell surface internalization and liposome delivery in cell culture.

    Science.gov (United States)

    Trubetskaya, O V; Trubetskoy, V S; Domogatsky, S P; Rudin, A V; Popov, N V; Danilov, S M; Nikolayeva, M N; Klibanov, A L; Torchilin, V P

    1988-02-01

    A monoclonal antibody (mAb), E25, is described that binds to the surface of cultured human endothelial cells. Upon binding E25 is rapidly internalized and digested intracellularly. Selective liposome targeting to the surface of the cells is performed using a biotinylated E25 antibody and an avidin-biotin system. Up to 30% of the cell-adherent liposomal lipid is internalized.

  20. Saliva analysis combining membrane protein purification with surface-enhanced Raman spectroscopy for nasopharyngeal cancer detection

    Science.gov (United States)

    Feng, Shangyuan; Lin, Duo; Lin, Juqiang; Huang, Zufang; Chen, Guannan; Li, Yongzeng; Huang, Shaohua; Zhao, Jianhua; Chen, Rong; Zeng, Haishan

    2014-02-01

    A method for saliva analysis combining membrane protein purification with silver nanoparticle-based surface-enhanced Raman spectroscopy (SERS) for non-invasive nasopharyngeal cancer detection was present in this paper. In this method, cellulose acetate membrane was used to obtain purified whole proteins from human saliva while removing other native saliva constituents and exogenous substances. The purified proteins were mixed with silver nanoparticle for SERS analysis. A diagnostic accuracy of 90.2% can be achieved by principal components analysis combined with linear discriminate analysis, for saliva samples obtained from patients with nasopharyngeal cancer (n = 62) and healthy volunteers (n = 30). This exploratory study demonstrated the potential for developing non-invasive, rapid saliva SERS analysis for nasopharyngeal cancer detection.

  1. Additive manufactured polymeric 3D scaffolds with tailored surface topography influence mesenchymal stromal cells activity.

    Science.gov (United States)

    Neves, Sara C; Mota, Carlos; Longoni, Alessia; Barrias, Cristina C; Granja, Pedro L; Moroni, Lorenzo

    2016-06-01

    Additive manufactured three-dimensional (3D) scaffolds with tailored surface topography constitute a clear advantage in tissue regeneration strategies to steer cell behavior. 3D fibrous scaffolds of poly(ethylene oxide terephthalate)/poly(butylene terephthalate) block copolymer presenting different fiber surface features were successfully fabricated by additive manufacturing combined with wet-spinning, in a single step, without any post-processing. The optimization of the processing parameters, mainly driven by different solvent/non-solvent combinations, led to four distinct scaffold types, with average surface roughness values ranging from 0.071 ± 0.012 μm to 1.950 ± 0.553 μm, average pore sizes in the x- and y-axis between 351.1 ± 33.6 μm and 396.1 ± 32.3 μm, in the z-axis between 36.5 ± 5.3 μm and 70.7 ± 8.8 μm, average fiber diameters between 69.4 ± 6.1 μm and 99.0 ± 9.4 μm, and porosity values ranging from 60.2 ± 0.8% to 71.7 ± 2.6%. Human mesenchymal stromal cells (hMSCs) cultured on these scaffolds adhered, proliferated, and produced endogenous extracellular matrix. The effect of surface roughness and topography on hMSCs differentiation was more evident for cells seeded at lower density, where the percentage of cells in direct contact with the surface was higher compared to more densely seeded scaffolds. Under osteogenic conditions, lower surface roughness values (0.227 ± 0.035 μm) had a synergistic effect on hMSCs behavior, while chondrogenesis was favored on rougher surfaces (1.950 ± 0.553 μm). PMID:27219645

  2. Method for Constructing Composite Response Surfaces by Combining Neural Networks with Polynominal Interpolation or Estimation Techniques

    Science.gov (United States)

    Rai, Man Mohan (Inventor); Madavan, Nateri K. (Inventor)

    2007-01-01

    A method and system for data modeling that incorporates the advantages of both traditional response surface methodology (RSM) and neural networks is disclosed. The invention partitions the parameters into a first set of s simple parameters, where observable data are expressible as low order polynomials, and c complex parameters that reflect more complicated variation of the observed data. Variation of the data with the simple parameters is modeled using polynomials; and variation of the data with the complex parameters at each vertex is analyzed using a neural network. Variations with the simple parameters and with the complex parameters are expressed using a first sequence of shape functions and a second sequence of neural network functions. The first and second sequences are multiplicatively combined to form a composite response surface, dependent upon the parameter values, that can be used to identify an accurate mode

  3. Surface-enhanced Raman spectroscopy of the endothelial cell membrane.

    Directory of Open Access Journals (Sweden)

    Simon W Fogarty

    Full Text Available We applied surface-enhanced Raman spectroscopy (SERS to cationic gold-labeled endothelial cells to derive SERS-enhanced spectra of the bimolecular makeup of the plasma membrane. A two-step protocol with cationic charged gold nanoparticles followed by silver-intensification to generate silver nanoparticles on the cell surface was employed. This protocol of post-labelling silver-intensification facilitates the collection of SERS-enhanced spectra from the cell membrane without contribution from conjugated antibodies or other molecules. This approach generated a 100-fold SERS-enhancement of the spectral signal. The SERS spectra exhibited many vibrational peaks that can be assigned to components of the cell membrane. We were able to carry out spectral mapping using some of the enhanced wavenumbers. Significantly, the spectral maps suggest the distribution of some membrane components are was not evenly distributed over the cells plasma membrane. These results provide some possible evidence for the existence of lipid rafts in the plasma membrane and show that SERS has great potential for the study and characterization of cell surfaces.

  4. Opto-mechanical probe for combining atomic force microscopy and optical near-field surface analysis

    OpenAIRE

    Hoorn, van der, T.; Chavan, D.C.; B. Tiribilli; Margheri, G.; Mank, A.J.G.; Ariese, F.; Iannuzzi, D.

    2014-01-01

    We have developed a new easy-to-use probe that can be used to combine atomic force microscopy (AFM) and scanning near-field optical microscopy (SNOM). We show that, using this device, the evanescent field, obtained by total internal reflection conditions in a prism, can be visualized by approaching the surface with the scanning tip. Furthermore, we were able to obtain simultaneous AFM and SNOM images of a standard test grating in air and in liquid. The lateral resolution in AFM and SNOM mode ...

  5. Adsorption of mercury on lignin: Combined surface complexation modeling and X-ray absorption spectroscopy studies

    International Nuclear Information System (INIS)

    Adsorption of mercury (Hg) on lignin was studied at a range of pH values using a combination of batch adsorption experiments, a surface complexation model (SCM) and synchrotron X-ray absorption spectroscopy (XAS). Surface complexation modeling indicates that three types of acid sites on lignin surfaces, namely aliphatic carboxylic-, aromatic carboxylic- and phenolic-type surface groups, contributed to Hg(II) adsorption. The bond distance and coordination number of Hg(II) adsorption samples at pH 3.0, 4.0 and 5.5 were obtained from extended X-ray absorption fine structure (EXAFS) spectroscopy analysis. The results of SCM and XAS combined reveal that the predominant adsorption species of Hg(II) on lignin changes from HgCl20 to monodentate complex –C–O–HgCl and then bidentate complex –C–O–Hg–O–C– with increasing pH value from 2.0 to 6.0. The good agreement between SCM and XAS results provides new insight into understanding the mechanisms of Hg(II) adsorption on lignin. - Highlights: ► Lignin exhibits a high Hg(II) adsorption capability. ► Adsorption of Hg(II) on lignin is strongly pH-dependent. ► HgCl20, –C–O–HgCl and –C–O–Hg–O–C– are the main adsorption species of Hg(II). - Adsorption of Hg(II) on lignin is strongly pH-dependent, and the main adsorption Hg(II) species change from HgCl20 to –C–O–HgCl and –C–O–Hg–O–C– as pH increases from 2.0 to 6.0.

  6. Combined experimental and theoretical investigation of interactions between kaolinite inner surface and intercalated dimethyl sulfoxide

    International Nuclear Information System (INIS)

    Graphical abstract: Snapshot of the kaolinite–DMSO system after equilibrium is reached. - Highlights: • Dimethyl sulfoxide arranges a monolayer structure between kaolinite layers. • Weak hydrogen bonds exist between methyl groups of dimethyl sulfoxide and kaolinite silica layer. • Intercalated dimethyl sulfoxide forms strong hydrogen bonds with kaolinite alumina layer. - Abstract: Kaolinite intercalation complex with dimethyl sulfoxide (DMSO) was investigated by X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), and thermogravimetry–differential scanning calorimetry (TG–DSC) combined with molecular dynamics simulation. The bands assigned to the OH stretching of inner surface of kaolinite were significantly perturbed after intercalation of DMSO into kaolinite. Additionally, the bands attributed to the vibration of gibbsite-like layers of kaolinite shifted to the lower wave number, indicating that the intercalated DMSO were strongly hydrogen bonded to the alumina octahedral surface of kaolinite. The slightly decreased intensity of 1031 cm−1 and 1016 cm−1 band due to the in-plane vibration of Si−O of kaolinite revealed that some DMSO molecules formed weak hydrogen bonds with the silicon tetrahedral surface of kaolinite. Based on the TG result of kaolinite–DMSO intercalation complex, the formula of A12Si2O5(OH)4(DMSO)0.7 was obtained, with which the kaolinite–DMSO complex model was constructed. The molecular dynamics simulation of kaolinite–DMSO complex directly confirmed the monolayer structure of DMSO in interlayer space of kaolinite, where the DMSO arranged almost parallel with kaolinite basal surface with all methyl groups being distributed near the interlayer midplane and oxygen atoms orienting toward to the alumina octahedral surface. The radial distribution function between kaolinite and intercalated DMSO verified the strong hydrogen bonds forming between hydroxyl hydrogen atoms of alumina octahedral surface and

  7. Combined experimental and theoretical investigation of interactions between kaolinite inner surface and intercalated dimethyl sulfoxide

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Shuai [School of Geoscience and Surveying Engineering, China University of Mining & Technology, Beijing 100083 (China); Liu, Qinfu, E-mail: lqf@cumtb.edu.cn [School of Geoscience and Surveying Engineering, China University of Mining & Technology, Beijing 100083 (China); Cheng, Hongfei [School of Geoscience and Surveying Engineering, China University of Mining & Technology, Beijing 100083 (China); Zeng, Fangui [Department of Earth Science and Engineering, Taiyuan University of Technology, Taiyuan 030024 (China)

    2015-03-15

    Graphical abstract: Snapshot of the kaolinite–DMSO system after equilibrium is reached. - Highlights: • Dimethyl sulfoxide arranges a monolayer structure between kaolinite layers. • Weak hydrogen bonds exist between methyl groups of dimethyl sulfoxide and kaolinite silica layer. • Intercalated dimethyl sulfoxide forms strong hydrogen bonds with kaolinite alumina layer. - Abstract: Kaolinite intercalation complex with dimethyl sulfoxide (DMSO) was investigated by X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), and thermogravimetry–differential scanning calorimetry (TG–DSC) combined with molecular dynamics simulation. The bands assigned to the OH stretching of inner surface of kaolinite were significantly perturbed after intercalation of DMSO into kaolinite. Additionally, the bands attributed to the vibration of gibbsite-like layers of kaolinite shifted to the lower wave number, indicating that the intercalated DMSO were strongly hydrogen bonded to the alumina octahedral surface of kaolinite. The slightly decreased intensity of 1031 cm{sup −1} and 1016 cm{sup −1} band due to the in-plane vibration of Si−O of kaolinite revealed that some DMSO molecules formed weak hydrogen bonds with the silicon tetrahedral surface of kaolinite. Based on the TG result of kaolinite–DMSO intercalation complex, the formula of A1{sub 2}Si{sub 2}O{sub 5}(OH){sub 4}(DMSO){sub 0.7} was obtained, with which the kaolinite–DMSO complex model was constructed. The molecular dynamics simulation of kaolinite–DMSO complex directly confirmed the monolayer structure of DMSO in interlayer space of kaolinite, where the DMSO arranged almost parallel with kaolinite basal surface with all methyl groups being distributed near the interlayer midplane and oxygen atoms orienting toward to the alumina octahedral surface. The radial distribution function between kaolinite and intercalated DMSO verified the strong hydrogen bonds forming between hydroxyl hydrogen

  8. Identification of polymer surface adsorbed proteins implicated in pluripotent human embryonic stem cell expansion.

    Science.gov (United States)

    Hammad, Moamen; Rao, Wei; Smith, James G W; Anderson, Daniel G; Langer, Robert; Young, Lorraine E; Barrett, David A; Davies, Martyn C; Denning, Chris; Alexander, Morgan R

    2016-08-16

    Improved biomaterials are required for application in regenerative medicine, biosensing, and as medical devices. The response of cells to the chemistry of polymers cultured in media is generally regarded as being dominated by proteins adsorbed to the surface. Here we use mass spectrometry to identify proteins adsorbed from a complex mouse embryonic fibroblast (MEF) conditioned medium found to support pluripotent human embryonic stem cell (hESC) expansion on a plasma etched tissue culture polystyrene surface. A total of 71 proteins were identified, of which 14 uniquely correlated with the surface on which pluripotent stem cell expansion was achieved. We have developed a microarray combinatorial protein spotting approach to test the potential of these 14 proteins to support expansion of a hESC cell line (HUES-7) and a human induced pluripotent stem cell line (ReBl-PAT) on a novel polymer (N-(4-Hydroxyphenyl) methacrylamide). These proteins were spotted to form a primary array yielding several protein mixture 'hits' that enhanced cell attachment to the polymer. A second array was generated to test the function of a refined set of protein mixtures. We found that a combination of heat shock protein 90 and heat shock protein-1 encourage elevated adherence of pluripotent stem cells at a level comparable to fibronectin pre-treatment. PMID:27466628

  9. Surface plasmon resonance imaging of cells and surface-associated fibronectin

    Directory of Open Access Journals (Sweden)

    Bhadriraju Kiran

    2009-02-01

    Full Text Available Abstract Background A critical challenge in cell biology is quantifying the interactions of cells with their extracellular matrix (ECM environment and the active remodeling by cells of their ECM. Fluorescence microscopy is a commonly employed technique for examining cell-matrix interactions. A label-free imaging method would provide an alternative that would eliminate the requirement of transfected cells and modified biological molecules, and if collected nondestructively, would allow long term observation and analysis of live cells. Results Using surface plasmon resonance imaging (SPRI, the deposition of protein by vascular smooth muscle cells (vSMC cultured on fibronectin was quantified as a function of cell density and distance from the cell periphery. We observed that as much as 120 ng/cm2 of protein was deposited by cells in 24 h. Conclusion SPRI is a real-time, low-light-level, label-free imaging technique that allows the simultaneous observation and quantification of protein layers and cellular features. This technique is compatible with live cells such that it is possible to monitor cellular modifications to the extracellular matrix in real-time.

  10. Mild hypothermia combined with neural stem cell transplantation for hypoxic-ischemic encephalopathy: neuroprotective effects of combined therapy

    OpenAIRE

    Lin WANG; Jiang, Feng; Li, Qifeng; He, Xiaoguang; Ma, Jie

    2014-01-01

    Neural stem cell transplantation is a useful treatment for ischemic stroke, but apoptosis often occurs in the hypoxic-ischemic environment of the brain after cell transplantation. In this study, we determined if mild hypothermia (27–28°C) can increase the survival rate of neural stem cells (1.0 × 105/μL) transplanted into neonatal mice with hypoxic-ischemic encephalopathy. Long-term effects on neurological functioning of the mice were also examined. After mild hypothermia combined with neural...

  11. Genotoxicity and cytotoxicity of cisplatin treatment combined with anaesthetics on EAT cells in vivo.

    Science.gov (United States)

    Brozovic, Gordana; Orsolic, Nada; Knezevic, Fabijan; Horvat Knezevic, Anica; Benkovic, Vesna; Sakic, Katarina; Hrgovic, Zlatko; Bendelja, Kreso; Fassbender, Walter J

    2009-06-01

    In this study, DNA damage in tumour cells, as well as irreversible cell damage leading to apoptosis induced in vivo by the combined application of cisplatin and inhalation anaesthetics, was investigated. The genotoxicity of anaesthetics on Ehrlich ascites tumour (EAT) cells of mice, alone or in combined application with cisplatin, was estimated by using the alkaline comet assay. The percentage of EAT cell apoptosis was quantified by flow cytometry. Groups of EAT-bearing mice were (i) treated intraperitoneally with cisplatin, (ii) exposed to repeated anaesthesia with inhalation anaesthetic, and (iii) subjected to combined treatment of exposure to anaesthetics after cisplatin for 3 days. Sevoflurane, halothane and isoflurane caused strong genotoxic effects on tumour cells in vivo. The tested anaesthetics alone showed no direct effect on programmed cell death although sevoflurane and especially halothane decreased the number of living EAT cells in peritoneal cavity lavage. Repeated anaesthesia with isoflurane had stimulatory effects on EAT cell proliferation and inhibited tumour cell apoptosis (6.11%), compared to the control group (10.26%). Cisplatin caused massive apoptosis of EAT cells (41.14%) and decreased the number of living EAT cells in the peritoneal cavity. Combined cisplatin and isoflurane treatment additionally increased EAT cell apoptosis to 51.32%. Combined treatment of mice with cisplatin and all anaesthetics increased the number of living tumour cells in the peritoneal cavity compared to cisplatin treatment of mice alone. These results suggest that the inhalation of anaesthetics may protect tumour cells from the cisplatin-induced genotoxic and cytotoxic effects.

  12. STUDY ON GLYCOCONJUGATE CHANGES ON CELL SURFACE IN PROGRESSIVE DEVELOPMENT OF PULMONARY TUMOR

    Institute of Scientific and Technical Information of China (English)

    WANG Xiao-mei; SHAN Jun; CHEN Zhuo-huai

    2002-01-01

    Aim: To investigate glycoconjugate changes on the cell surface of proliferative lesions and neoplasms of mice lungs at various stages of tumorigenesis, the relation between progressive development of mouse pulmonary tumors and expression of cell surface saccharide. Materials and methods: Thirty - one male A/J strain mice at 5 weeks of age were treated intraperitoneally with a single injection of 20 - methylcholanthrene (20 - MC), 292 pulmonary lesions including 31 hyperplasias, 145 alveolar adenomas, 61 papillary adenomas, 55 papillary adenocarcinomas and their combined type were obtained. The binding affinities of cells in normal respiratory epithelia and in proliferative lesions to four peroxidases - conjugated lectins, Maclura pomifera agglutinin (MPA), Arachis hypogea agglutinin (PNA), Ricinus communis agglutinin (RCA), and wheat germ agglutinin (WGA) were examined. Results: Cells of hyperplasia and alveolar adenoma showed fairly strong affinity to all the four lectins. However, part of papillary adenoma cells and greater part of papillary adenocarcinoma cells lost their binding affinity to MPA, PNA, and RCA, but not to WGA. The bindings of MPA, PNA and RNA were detected predominently on the luminal surfaces of benign tumors but not on the luminal surfaces of malignant tumors. WGA might bind to varied types of benign and malignant tumors. Pretreated with neuraminidase, the lesions enhanced the staining intensity for the four lectins, the binding sites of WGA to malignant tumor cells were numerous. A distinct difference in lectin binding affinity between hyperplasia / alveolar adenoma/papillary adenoma and papillary adenocarcinoma was clearly shown( x2 = 46.89, P < 0.01, x2 = 36.77, P < 0.01 and x2 = 52.87, P < 0.01 ) in this experiment. The complex glycoconjugates on the cell surface of malignant and benign lesions during the development of pulmonary tumor were changed,malignant tumor cells differed from the surface of benign tumor cells, the levels of

  13. Multijunction Solar Cells Optimized for the Mars Surface Solar Spectrum

    Science.gov (United States)

    Edmondson, Kenneth M.; Fetzer, Chris; Karam, Nasser H.; Stella, Paul; Mardesich, Nick; Mueller, Robert

    2007-01-01

    This paper gives an update on the performance of the Mars Exploration Rovers (MER) which have been continually performing for more than 3 years beyond their original 90-day missions. The paper also gives the latest results on the optimization of a multijunction solar cell that is optimized to give more power on the surface of Mars.

  14. Bacterial Cell Surface Damage Due to Centrifugal Compaction

    NARCIS (Netherlands)

    Peterson, Brandon W.; Sharma, Prashant K.; van der Mei, Henny C.; Busscher, Henk J.

    2012-01-01

    Centrifugal damage has been known to alter bacterial cell surface properties and interior structures, including DNA. Very few studies exist on bacterial damage caused by centrifugation because of the difficulty in relating centrifugation speed and container geometry to the damage caused. Here, we pr

  15. Functional recoveries of sciatic nerve regeneration by combining chitosan-coated conduit and neurosphere cells induced from adipose-derived stem cells.

    Science.gov (United States)

    Hsueh, Yuan-Yu; Chang, Ya-Ju; Huang, Tzu-Chieh; Fan, Shih-Chen; Wang, Duo-Hsiang; Chen, Jia-Jin Jason; Wu, Chia-Ching; Lin, Sheng-Che

    2014-02-01

    Suboptimal repair occurs in a peripheral nerve gap, which can be partially restored by bridging the gap with various biosynthetic conduits or cell-based therapy. In this study, we developed a combination of chitosan coating approach to induce neurosphere cells from human adipose-derived stem cells (ASCs) on chitosan-coated plate and then applied these cells to the interior of a chitosan-coated silicone tube to bridge a 10-mm gap in a rat sciatic nerve. Myelin sheath degeneration and glial scar formation were discovered in the nerve bridged by the silicone conduit. By using a single treatment of chitosan-coated conduit or neurosphere cell therapy, the nerve gap was partially recovered after 6 weeks of surgery. Substantial improvements in nerve regeneration were achieved by combining neurosphere cells and chitosan-coated conduit based on the increase of myelinated axons density and myelin thickness, gastrocnemius muscle weight and muscle fiber diameter, and step and stride lengths from gait analysis. High expressions of interleukin-1β and leukotriene B4 receptor 1 in the intra-neural scarring caused by using silicone conduits revealed that the inflammatory mechanism can be inhibited when the conduit is coated with chitosan. This study demonstrated that the chitosan-coated surface performs multiple functions that can be used to induce neurosphere cells from ASCs and to facilitate nerve regeneration in combination with a cells-assisted coated conduit. PMID:24360575

  16. Cell surface hydrophobicity is conveyed by S-layer proteins - A study in recombinant lactobacilli

    NARCIS (Netherlands)

    Mei, H.C. van der; Belt-Gritter, B. van de; Pouwels, P.H.; Martinez, B.; Busscher, H.J.

    2003-01-01

    Cell surface hydrophobicity is one of the most important factors controlling adhesion of microorganisms to surfaces. In this paper, cell surface properties of lactobacilli and recombinant lactobacilli with and without a surface layer protein (SLP) associated with cell surface hydrophobicity were det

  17. Strengthening of RC Beams Using Externally Bonded Reinforcement Combined with Near-Surface Mounted Technique

    Directory of Open Access Journals (Sweden)

    Kh Mahfuz ud Darain

    2016-07-01

    Full Text Available This study investigates the flexural behaviour of reinforced concrete (RC beams strengthened through the combined externally bonded and near-surface mounted (CEBNSM technique. The externally bonded reinforcement (EBR and near-surface mounted (NSM techniques are popular strengthening solutions, although these methods often demonstrate premature debonding failure. The proposed CEBNSM technique increases the bond area of the concrete–carbon fibre reinforced polymer (CFRP interface, which can delay the debonding failure. This technique is appropriate when any structure has a narrow cross-sectional width or is in need of additional flexural capacity that an individual technique or material cannot attain. An experimental test matrix was designed with one control and five strengthened RC beams to verify the performance of the proposed technique. The strengthening materials were CFRP bar as NSM reinforcement combined with CFRP fabric as EBR material. The test variables were the diameter of the NSM bars (8 and 10 mm, the thickness of the CFRP fabrics (one and two layers and the U-wrap anchorage. The strengthened beams showed enhancement of ultimate load capacity, stiffness, cracking behaviour, and strain compatibility. The ultimate capacity of the CEBNSM-strengthened beams increased from 71% to 105% compared to that of the control beam. A simulation method based on the moment-rotation approach was also presented to predict the behaviour of CEBNSM-strengthened RC beams.

  18. Cell Recovery after Combined Action of Ionizing Radiation and Chemicals

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Jin Kyu; Roh, Chang Hyun; Ryu, Tae Ho [Korea Atomic Energy Research Institute, Jeongeup (Korea, Republic of); Komarova, Ludmila N.; Petin, Vladislav G. [Medical Radiological Research Center, Obninsk (Russian Federation)

    2012-05-15

    Damage repair in malignant cells would be problematic in sterilization of microorganisms and treatment of cancer, as well. The inhibition of cell recovery and DNA single and double strand breaks repair by chemicals is expressed both as a deceleration of recovery rate and a lesser extent of recovery. Three possibilities are involved in the inhibition of cell recovery: (1) impairment of the recovery process itself, (2) increased irreversible damage, and (3) simultaneous exert of the two. There have been fee publications regarding these problems. The aim of this study was to determine which of these points are involved in the inhibition of cell recovery. In this study, a quantitative approach describing cell recovery from potentially lethal damage as a decrease in the effective dose was used

  19. Methods To Identify Aptamers against Cell Surface Biomarkers

    Directory of Open Access Journals (Sweden)

    Frédéric Ducongé

    2011-09-01

    Full Text Available Aptamers are nucleic acid-based ligands identified through a process of molecular evolution named SELEX (Systematic Evolution of Ligands by Exponential enrichment. During the last 10-15 years, numerous aptamers have been developed specifically against targets present on or associated with the surface of human cells or infectious pathogens such as viruses, bacteria, fungi or parasites. Several of the aptamers have been described as potent probes, rivalling antibodies, for use in flow cytometry or microscopy. Some have also been used as drugs by inhibiting or activating functions of their targets in a manner similar to neutralizing or agonistic antibodies. Additionally, it is straightforward to conjugate aptamers to other agents without losing their affinity and they have successfully been used in vitro and in vivo to deliver drugs, siRNA, nanoparticles or contrast agents to target cells. Hence, aptamers identified against cell surface biomarkers represent a promising class of ligands. This review presents the different strategies of SELEX that have been developed to identify aptamers for cell surface-associated proteins as well as some of the methods that are used to study their binding on living cells.

  20. Longitudinal microarray analysis of cell surface antigens on peripheral blood mononuclear cells from HIV+ individuals on highly active antiretroviral therapy

    Directory of Open Access Journals (Sweden)

    Wang Bin

    2008-03-01

    Full Text Available Abstract Background The efficacy of highly active antiretroviral therapy (HAART determined by simultaneous monitoring over 100 cell-surface antigens overtime has not been attempted. We used an antibody microarray to analyze changes in the expression of 135 different cell-surface antigens overtime on PBMC from HIV+ patients on HAART. Two groups were chosen, one (n = 6 achieved sustainable response by maintaining below detectable plasma viremia and the other (n = 6 responded intermittently. Blood samples were collected over an average of 3 years and 5–8 time points were selected for microarray assay and statistical analysis. Results Significant trends over time were observed for the expression of 7 cell surface antigens (CD2, CD3epsilon, CD5, CD95, CD36, CD27 and CD28 for combined patient groups. Between groups, expression levels of 10 cell surface antigens (CD11a, CD29, CD38, CD45RO, CD52, CD56, CD57, CD62E, CD64 and CD33 were found to be differential. Expression levels of CD9, CD11a, CD27, CD28 and CD52, CD44, CD49d, CD49e, CD11c strongly correlated with CD4+ and CD8+ T cell counts, respectively. Conclusion Our findings not only detected markers that may have potential prognostic/diagnostic values in evaluating HAART efficacy, but also showed how density of cell surface antigens could be efficiently exploited in an array-like manner in relation to HAART and HIV-infection. The antigens identified in this study should be further investigated by other methods such as flow cytometry for confirmation as biological analysis of these antigens may help further clarify their role during HAART and HIV infection.

  1. "Race for the Surface": Eukaryotic Cells Can Win.

    Science.gov (United States)

    Pham, Vy T H; Truong, Vi Khanh; Orlowska, Anna; Ghanaati, Shahram; Barbeck, Mike; Booms, Patrick; Fulcher, Alex J; Bhadra, Chris M; Buividas, Ričardas; Baulin, Vladimir; Kirkpatrick, C James; Doran, Pauline; Mainwaring, David E; Juodkazis, Saulius; Crawford, Russell J; Ivanova, Elena P

    2016-08-31

    With an aging population and the consequent increasing use of medical implants, managing the possible infections arising from implant surgery remains a global challenge. Here, we demonstrate for the first time that a precise nanotopology provides an effective intervention in bacterial cocolonization enabling the proliferation of eukaryotic cells on a substratum surface, preinfected by both live Gram-negative, Pseudomonas aeruginosa, and Gram-positive, Staphylococcus aureus, pathogenic bacteria. The topology of the model black silicon (bSi) substratum not only favors the proliferation of eukaryotic cells but is biocompatible, not triggering an inflammatory response in the host. The attachment behavior and development of filopodia when COS-7 fibroblast cells are placed in contact with the bSi surface are demonstrated in the dynamic study, which is based on the use of real-time sequential confocal imaging. Bactericidal nanotopology may enhance the prospect for further development of inherently responsive antibacterial nanomaterials for bionic applications such as prosthetics and implants.

  2. Cell adhesion on Ti surface with controlled roughness

    Energy Technology Data Exchange (ETDEWEB)

    Burgos-Asperilla, L.; Garcia-Alonso, M. C.; Escudero, M. L.; Alonso, C.

    2015-07-01

    In this report, the in situ interaction between Saos-2 osteoblast cells and a smooth Ti surface was examined over time. The adhesion kinetics and mechanisms of cellular proliferation were monitored by quartz crystal microbalance (QCM) and electrochemical impedance spectroscopy (EIS). The rate of Saos-2 attachment on Ti surfaces, obtained from the measurements performed with the QCM, is a first-order reaction, with k=2.10{sup -}3 min{sup -}1. The impedance measurements indicate that in the absence of cells, the Ti resistance diminishes over time (7 days), due to the presence of amino acids and proteins from the culture medium that have been adsorbed, while in the presence of osteoblasts, this decrease is much greater because of the compounds generated by the cells that accelerate the dissolution of Ti. (Author)

  3. Surgery combined with topical photodynamic therapy for the treatment of squamous cell carcinoma of the lip.

    Science.gov (United States)

    Wang, Yuanyuan; Yang, Yadong; Yang, Yunchuan; Lu, Yuangang

    2016-06-01

    Due to the unique location of the squamous cell carcinoma (SCC) of the lip, using a single method such as extended resection or radiotherapy probably causes morphological and functional defects. So we used surgery combined with topical photodynamic therapy (PDT) to treat SCC of the lip. Under local anesthesia with 5% lidocaine, the hyperplastic and ulcerative SCC of the lip were curetted and assisted by topical PDTs after surgery. The 20% 5-aminolevulinic acid cream was used as a photosensitizer and applied evenly to the surface of the tumor lesion for 4h. Then the lesion site was irradiated with a 635-nm laser at 120J/cm(2). A total of five PDTs were performed postoperatively at an interval of 2 weeks. Photos were taken before and after every PDT to compare the skin lesions, treatment effects, and side effects. A long-term follow-up was undertaken to observe tumor recurrence. After surgery combined with five topical PDTs, the SCC of the lip disappeared without the compromised morphology of the lip, significant side effects, or tumor recurrence in one-year follow-up. Surgery combined with topical PDT can reduce the excision size of tumors and play a positive role in the treatment of tumors of special locations.

  4. Galectin-1-mediated cell adhesion, invasion and cell death in human anaplastic large cell lymphoma: Regulatory roles of cell surface glycans

    OpenAIRE

    Suzuki, Osamu; Abe, Masafumi

    2014-01-01

    Galectin-1 is known to be one of the extracellular matrix proteins. To elucidate the biological roles of galectin-1 in cell adhesion and invasion of human anaplastic large cell lymphoma, we performed cell adhesion and invasion assays using the anaplastic large cell lymphoma cell line H-ALCL, which was previously established in our laboratory. From the cell surface lectin array, treatment with neuraminidase from Arthrobacter ureafaciens which cleaves all linkage types of cell surface sialic ac...

  5. Surface modification of hydrophobic polymers for improvement of endothelial cell-surface interactions

    NARCIS (Netherlands)

    Dekker, A.; Reitsma, K.; Beugeling, T.; Bantjes, A.; Feijen, J.; Kirkpatrick, C.J.; Aken, van W.G.

    1992-01-01

    The aim of this study is to improve the interaction of endothelial cells with polymers used in vascular prostheses. Polytetrafluoroethylene (PTFE; Teflon) films were treated by means of nitrogen and oxygen plasmas. Depending on the plasma exposure time, modified PTFE surfaces showed water-contact an

  6. Full skin quantitative optical coherence elastography achieved by combining vibration and surface acoustic wave methods

    Science.gov (United States)

    Li, Chunhui; Guan, Guangying; Huang, Zhihong; Wang, Ruikang K.; Nabi, Ghulam

    2015-03-01

    By combining with the phase sensitive optical coherence tomography (PhS-OCT), vibration and surface acoustic wave (SAW) methods have been reported to provide elastography of skin tissue respectively. However, neither of these two methods can provide the elastography in full skin depth in current systems. This paper presents a feasibility study on an optical coherence elastography method which combines both vibration and SAW in order to give the quantitative mechanical properties of skin tissue with full depth range, including epidermis, dermis and subcutaneous fat. Experiments are carried out on layered tissue mimicking phantoms and in vivo human forearm and palm skin. A ring actuator generates vibration while a line actuator were used to excited SAWs. A PhS-OCT system is employed to provide the ultrahigh sensitive measurement of the generated waves. The experimental results demonstrate that by the combination of vibration and SAW method the full skin bulk mechanical properties can be quantitatively measured and further the elastography can be obtained with a sensing depth from ~0mm to ~4mm. This method is promising to apply in clinics where the quantitative elasticity of localized skin diseases is needed to aid the diagnosis and treatment.

  7. A molecular smart surface for spatio-temporal studies of cell mobility.

    Directory of Open Access Journals (Sweden)

    Eun-ju Lee

    Full Text Available Active migration in both healthy and malignant cells requires the integration of information derived from soluble signaling molecules with positional information gained from interactions with the extracellular matrix and with other cells. How a cell responds and moves involves complex signaling cascades that guide the directional functions of the cytoskeleton as well as the synthesis and release of proteases that facilitate movement through tissues. The biochemical events of the signaling cascades occur in a spatially and temporally coordinated manner then dynamically shape the cytoskeleton in specific subcellular regions. Therefore, cell migration and invasion involve a precise but constantly changing subcellular nano-architecture. A multidisciplinary effort that combines new surface chemistry and cell biological tools is required to understand the reorganization of cytoskeleton triggered by complex signaling during migration. Here we generate a class of model substrates that modulate the dynamic environment for a variety of cell adhesion and migration experiments. In particular, we use these dynamic substrates to probe in real-time how the interplay between the population of cells, the initial pattern geometry, ligand density, ligand affinity and integrin composition affects cell migration and growth. Whole genome microarray analysis indicates that several classes of genes ranging from signal transduction to cytoskeletal reorganization are differentially regulated depending on the nature of the surface conditions.

  8. A molecular smart surface for spatio-temporal studies of cell mobility.

    Science.gov (United States)

    Lee, Eun-ju; Luo, Wei; Chan, Eugene W L; Yousaf, Muhammad N

    2015-01-01

    Active migration in both healthy and malignant cells requires the integration of information derived from soluble signaling molecules with positional information gained from interactions with the extracellular matrix and with other cells. How a cell responds and moves involves complex signaling cascades that guide the directional functions of the cytoskeleton as well as the synthesis and release of proteases that facilitate movement through tissues. The biochemical events of the signaling cascades occur in a spatially and temporally coordinated manner then dynamically shape the cytoskeleton in specific subcellular regions. Therefore, cell migration and invasion involve a precise but constantly changing subcellular nano-architecture. A multidisciplinary effort that combines new surface chemistry and cell biological tools is required to understand the reorganization of cytoskeleton triggered by complex signaling during migration. Here we generate a class of model substrates that modulate the dynamic environment for a variety of cell adhesion and migration experiments. In particular, we use these dynamic substrates to probe in real-time how the interplay between the population of cells, the initial pattern geometry, ligand density, ligand affinity and integrin composition affects cell migration and growth. Whole genome microarray analysis indicates that several classes of genes ranging from signal transduction to cytoskeletal reorganization are differentially regulated depending on the nature of the surface conditions. PMID:26030281

  9. Adhesion of MRC-5 and A549 cells on poly(dimethylsiloxane) surface modified by proteins.

    Science.gov (United States)

    Zuchowska, Agnieszka; Kwiatkowski, Piotr; Jastrzebska, Elzbieta; Chudy, Michal; Dybko, Artur; Brzozka, Zbigniew

    2016-02-01

    PDMS is a very popular material used for fabrication of Lab-on-a-Chip systems for biological applications. Although PDMS has numerous advantages, it is a highly hydrophobic material, which inhibits adhesion and proliferation of the cells. PDMS surface modifications are used to enrich growth of the cells. However, due to the fact that each cell type has specific adhesion, it is necessary to optimize the parameters of these modifications. In this paper, we present an investigation of normal (MRC-5) and carcinoma (A549) human lung cell adhesion and proliferation on modified PDMS surfaces. We have chosen these cell types because often they are used as models for basic cancer research. To the best of our knowledge, this is the first presentation of this type of investigation. The combination of a gas-phase processing (oxygen plasma or ultraviolet irradiation) and wet chemical methods based on proteins' adsorption was used in our experiments. Different proteins such as poly-l-lysine, fibronectin, laminin, gelatin, and collagen were incubated with the activated PDMS samples. To compare with other works, here, we also examined how ratio of prepolymer to curing agent (5:1, 10:1, and 20:1) influences PDMS hydrophilicity during further modifications. The highest adhesion of the tested cells was observed for the usage of collagen, regardless of PDMS ratio. However, the MRC-5 cell line demonstrated better adhesion than A549 cells. This is probably due to the difference in their morphology and type (normal/cancer). PMID:26311334

  10. Recognition and prediction of individual and combined muscular activation modes via surface EMG analysis

    Directory of Open Access Journals (Sweden)

    Daniel Graupe

    2010-09-01

    Full Text Available The paper discusses how recognition of individual and combined muscular activation modes (functions and the prediction of intended such modes can be accomplished by identifying parameters of noninvasive surface EMG signals. It outlines the mathematical analysis of surface EMG signal to facilitate such recognition and related prediction, including recognition of intention (in terms of attempts to activate motor functions from the EMG, without accessing the CNS itself, in cases where a patient, say, a high-level amputee does not have the final-activation muscles and joints. The EMG activity thus allows to interpret and recognize CNS commands from minute variations in the parameters of surface EMG signals that record changes in the firing of motor neurons triggering contractions in related muscle fibers. We note that although in popular media this is sometimes referred to as detection of “thoughts”, no thoughts are detected, but only motor-outcomes of thoughts as found in the EMG signal. Examples of concrete cases where such recognition or prediction were accomplished in the author’s lab and in devices that came out of that lab, are given as are references to these in the literature over the last 35 years.

  11. Bionic design for surface optimization combining hydrophilic and negative charged biological macromolecules.

    Science.gov (United States)

    Ran, Fen; Song, Haiming; Niu, Xiaoqin; Yang, Aimei; Nie, Shengqiang; Wang, Lingren; Li, Jie; Sun, Shudong; Zhao, Changsheng

    2014-06-01

    While polyethersulfone (PES) membrane represents a promising option for blood purification, the blood compatibility must be dramatically enhanced to meet today's ever-increasing demands for many emerging application. In this study, we report a bionic design for optimization and development of a modified PES membrane combining hydrophilic and negative charged biological macromolecules on its surface. The hydrophilic and ionic charged biological macromolecules sulfonated poly(styrene)-b-poly(methyl methacrylate)-b-poly-(styrene) (PSSMSS) and poly(vinyl pyrrolidone)-b-poly(methyl methacrylate)-b-poly-(vinyl pyrrolidone) were synthesized via reversible addition-fragmentation chain transfer polymerization and used together to modify PES membranes by blending method. A hydrophilic membrane surface with negative charged surface coating was obtained, imitating the hydrophilic and negatively charged structure feature of heparin. The modified PES membranes showed suppressed platelet adhesion, and a prolonged blood clotting time, and thereby improved blood compatibility. In addition, the blood clotting time of the modified membranes increased with the blended PSSMSS amounts increment, indicating that both the hydrophilic and negative charged groups play important roles in improving the blood compatibility of PES membranes.

  12. Cell-surface expression of Hsp70 on hematopoietic cancer cells after inhibition of HDAC activity

    DEFF Research Database (Denmark)

    Jensen, Helle; Andresen, Lars; Hansen, Karen Aagaard;

    2009-01-01

    We show that inhibition of HDAC activity leads to surface expression of Hsp70 on various hematopoietic cancer cells, an occurance that was not observed on naïve or activated peripheral blood cells. HDAC inhibitor-mediated Hsp70 surface expression was confined to the apoptotic Annexin V-positive c......We show that inhibition of HDAC activity leads to surface expression of Hsp70 on various hematopoietic cancer cells, an occurance that was not observed on naïve or activated peripheral blood cells. HDAC inhibitor-mediated Hsp70 surface expression was confined to the apoptotic Annexin V......-positive cells and blocked by inhibition of apoptosis. Other chemotherapeutic inducers of apoptosis such as etoposide and camptothecin also led to a robust induction of Hsp70 surface expression. Hsp70 expression was, however, not caused by induction of apoptosis per se, as activated CD4 T cells remained Hsp70...... surface-negative despite effective induction of apoptosis. Interestingly, inhibition of endolysosomes or normal ER/Golgi transport did not affect Hsp70 surface expression. Intracellular calcium and the transcription factor Sp1, which has been shown previously to be important for the intracellular stress...

  13. MEMS-based dynamic cell-to-cell culture platforms using electrochemical surface modifications

    International Nuclear Information System (INIS)

    MEMS-based biological platforms with the capability of both spatial placements and time releases of living cells for cell-to-cell culture experiments have been designed and demonstrated utilizing electrochemical surface modification effects. The spatial placement is accomplished by electrochemical surface modification of substrate surfaces to be either adhesive or non-adhesive for living cells. The time control is achieved by the electrical activation of the selective indium tin oxide co-culture electrode to allow the migration of living cells onto the electrode to start the cell-to-cell culture studies. Prototype devices have a three-electrode design with an electrode size of 50 × 50 µm2 and the separation gaps of 2 µm between them. An electrical voltage of −1.5 V has been used to activate the electrodes independently and sequentially to demonstrate the dynamic cell-to-cell culture experiments of NIH 3T3 fibroblast and Madin Darby canine kidney cells. As such, this MEMS platform could be a basic yet versatile tool to characterize transient cell-to-cell interactions

  14. Proteomics and glycoproteomics of pluripotent stem-cell surface proteins.

    Science.gov (United States)

    Sun, Bingyun

    2015-03-01

    Pluripotent stem cells are a unique cell type with promising potential in regenerative and personalized medicine. Yet the difficulty to understand and coax their seemingly stochastic differentiation and spontaneous self-renewal have largely limited their clinical applications. A call has been made by numerous researchers for a better characterization of surface proteins on these cells, in search of biomarkers that can dictate developmental stages and lineage specifications, and can help formulate mechanistic insight of stem-cell fate choices. In the past two decades, proteomics has gained significant recognition in profiling surface proteins at high throughput. This review will summarize the impact of these studies on stem-cell biology, and discuss the used proteomic techniques. A systematic comparison of all the techniques and their results is also attempted here to help reveal pros, cons, and the complementarity of the existing methods. This awareness should assist in selecting suitable strategies for stem-cell related research, and shed light on technical improvements that can be explored in the future. PMID:25211708

  15. Proteomics and glycoproteomics of pluripotent stem-cell surface proteins.

    Science.gov (United States)

    Sun, Bingyun

    2015-03-01

    Pluripotent stem cells are a unique cell type with promising potential in regenerative and personalized medicine. Yet the difficulty to understand and coax their seemingly stochastic differentiation and spontaneous self-renewal have largely limited their clinical applications. A call has been made by numerous researchers for a better characterization of surface proteins on these cells, in search of biomarkers that can dictate developmental stages and lineage specifications, and can help formulate mechanistic insight of stem-cell fate choices. In the past two decades, proteomics has gained significant recognition in profiling surface proteins at high throughput. This review will summarize the impact of these studies on stem-cell biology, and discuss the used proteomic techniques. A systematic comparison of all the techniques and their results is also attempted here to help reveal pros, cons, and the complementarity of the existing methods. This awareness should assist in selecting suitable strategies for stem-cell related research, and shed light on technical improvements that can be explored in the future.

  16. Efficiency improvement of multicrystalline silicon solar cells after surface and grain boundaries passivation using vanadium oxide

    Energy Technology Data Exchange (ETDEWEB)

    Derbali, L., E-mail: rayan.slat@yahoo.fr [Photovoltaiec Laboratory, Research and Technology Center of Energy, Technopole de Borj-Cedria, BP 95, Hammam-Lif 2050 (Tunisia); Ezzaouia, H. [Photovoltaiec Laboratory, Research and Technology Center of Energy, Technopole de Borj-Cedria, BP 95, Hammam-Lif 2050 (Tunisia)

    2012-08-01

    Highlights: Black-Right-Pointing-Pointer Evaporation of vanadium pentoxide onto the front surface leads to reduce the surface reflectivity considerably. Black-Right-Pointing-Pointer An efficient surface passivation can be obtained after thermal treatment of obtained films. Black-Right-Pointing-Pointer Efficiency of the obtained solar cells has been improved noticeably after thermal treatment of deposited thin films. - Abstract: The aim of this work is to investigate the effect of vanadium oxide deposition onto the front surface of multicrystalline silicon (mc-Si) substrat, without any additional cost in the fabrication process and leading to an efficient surface and grain boundaries (GBs) passivation that have not been reported before. The lowest reflectance of mc-Si coated with vanadium oxide film of 9% was achieved by annealing the deposited film at 600 Degree-Sign C. Vanadium pentoxide (V{sub 2}O{sub 5}) were thermally evaporated onto the surface of mc-Si substrates, followed by a short annealing duration at a temperature ranging between 600 Degree-Sign C and 800 Degree-Sign C, under O{sub 2} atmosphere. The chemical composition of the films was analyzed by means of Fourier transform infrared spectroscopy (FTIR). Surface and cross-section morphology were determined by atomic force microscope (AFM) and a scanning electron microscope (SEM), respectively. The deposited vanadium oxide thin films make the possibility of combining in one processing step an antireflection coating deposition along with efficient surface state passivation, as compared to a reference wafer. Silicon solar cells based on untreated and treated mc-Si wafers were achieved. We showed that mc-silicon solar cells, subjected to the above treatment, have better short circuit currents and open-circuit voltages than those made from untreated wafers. Thus, the efficiency of obtained solar cells has been improved.

  17. Defined plant extracts can protect human cells against combined xenobiotic effects

    Directory of Open Access Journals (Sweden)

    Clair Emilie

    2011-01-01

    Full Text Available Abstract Background Pollutants representative of common environmental contaminants induce intracellular toxicity in human cells, which is generally amplified in combinations. We wanted to test the common pathways of intoxication and detoxification in human embryonic and liver cell lines. We used various pollutants such as Roundup residues, Bisphenol-A and Atrazine, and five precise medicinal plant extracts called Circ1, Dig1, Dig2, Sp1, and Uro1 in order to understand whether specific molecular actions took place or not. Methods Kidney and liver are major detoxification organs. We have studied embryonic kidney and hepatic human cell lines E293 and HepG2. The intoxication was induced on the one hand by a formulation of one of the most common herbicides worldwide, Roundup 450 GT+ (glyphosate and specific adjuvants, and on the other hand by a mixture of Bisphenol-A and Atrazine, all found in surface waters, feed and food. The prevention and curative effects of plant extracts were also measured on mitochondrial succinate dehydrogenase activity, on the entry of radiolabelled glyphosate (in Roundup in cells, and on cytochromes P450 1A2 and 3A4 as well as glutathione-S-transferase. Results Clear toxicities of pollutants were observed on both cell lines at very low sub-agricultural dilutions. The prevention of such phenomena took place within 48 h with the plant extracts tested, with success rates ranging between 25-34% for the E293 intoxicated by Roundup, and surprisingly up to 71% for the HepG2. By contrast, after intoxication, no plant extract was capable of restoring E293 viability within 48 h, however, two medicinal plant combinations did restore the Bisphenol-A/Atrazine intoxicated HepG2 up to 24-28%. The analysis of underlying mechanisms revealed that plant extracts were not capable of preventing radiolabelled glyphosate from entering cells; however Dig2 did restore the CYP1A2 activity disrupted by Roundup, and had only a mild preventive effect

  18. Surface code—biophysical signals for apoptotic cell clearance

    Science.gov (United States)

    Biermann, Mona; Maueröder, Christian; Brauner, Jan M.; Chaurio, Ricardo; Janko, Christina; Herrmann, Martin; Muñoz, Luis E.

    2013-12-01

    Apoptotic cell death and the clearance of dying cells play an important and physiological role in embryonic development and normal tissue turnover. In contrast to necrosis, apoptosis proceeds in an anti-inflammatory manner. It is orchestrated by the timed release and/or exposure of so-called ‘find-me’, ‘eat me’ and ‘tolerate me’ signals. Mononuclear phagocytes are attracted by various ‘find-me’ signals, including proteins, nucleotides, and phospholipids released by the dying cell, whereas the involvement of granulocytes is prevented via ‘stay away’ signals. The exposure of anionic phospholipids like phosphatidylserine (PS) by apoptotic cells on the outer leaflet of the plasma membrane is one of the main ‘eat me’ signals. PS is recognized by a number of innate receptors as well as by soluble bridging molecules on the surface of phagocytes. Importantly, phagocytes are able to discriminate between viable and apoptotic cells both exposing PS. Due to cytoskeleton remodeling PS has a higher lateral mobility on the surfaces of apoptotic cells thereby promoting receptor clustering on the phagocyte. PS not only plays an important role in the engulfment process, but also acts as ‘tolerate me’ signal inducing the release of anti-inflammatory cytokines by phagocytes. An efficient and fast clearance of apoptotic cells is required to prevent secondary necrosis and leakage of intracellular danger signals into the surrounding tissue. Failure or prolongation of the clearance process leads to the release of intracellular antigens into the periphery provoking inflammation and development of systemic inflammatory autoimmune disease like systemic lupus erythematosus. Here we review the current findings concerning apoptosis-inducing pathways, important players of apoptotic cell recognition and clearance as well as the role of membrane remodeling in the engulfment of apoptotic cells by phagocytes.

  19. Surface code—biophysical signals for apoptotic cell clearance

    International Nuclear Information System (INIS)

    Apoptotic cell death and the clearance of dying cells play an important and physiological role in embryonic development and normal tissue turnover. In contrast to necrosis, apoptosis proceeds in an anti-inflammatory manner. It is orchestrated by the timed release and/or exposure of so-called ‘find-me’, ‘eat me’ and ‘tolerate me’ signals. Mononuclear phagocytes are attracted by various ‘find-me’ signals, including proteins, nucleotides, and phospholipids released by the dying cell, whereas the involvement of granulocytes is prevented via ‘stay away’ signals. The exposure of anionic phospholipids like phosphatidylserine (PS) by apoptotic cells on the outer leaflet of the plasma membrane is one of the main ‘eat me’ signals. PS is recognized by a number of innate receptors as well as by soluble bridging molecules on the surface of phagocytes. Importantly, phagocytes are able to discriminate between viable and apoptotic cells both exposing PS. Due to cytoskeleton remodeling PS has a higher lateral mobility on the surfaces of apoptotic cells thereby promoting receptor clustering on the phagocyte. PS not only plays an important role in the engulfment process, but also acts as ‘tolerate me’ signal inducing the release of anti-inflammatory cytokines by phagocytes. An efficient and fast clearance of apoptotic cells is required to prevent secondary necrosis and leakage of intracellular danger signals into the surrounding tissue. Failure or prolongation of the clearance process leads to the release of intracellular antigens into the periphery provoking inflammation and development of systemic inflammatory autoimmune disease like systemic lupus erythematosus. Here we review the current findings concerning apoptosis-inducing pathways, important players of apoptotic cell recognition and clearance as well as the role of membrane remodeling in the engulfment of apoptotic cells by phagocytes. (paper)

  20. Involvement of cell surface phosphatidylinositol-anchored glycoproteins in cell-cell adhesion of chick embryo myoblasts

    OpenAIRE

    1989-01-01

    During myogenesis myoblasts fuse to form multinucleate cells that express muscle-specific proteins. A specific cell-cell adhesion process precedes lipid bilayer union during myoblast fusion (Knudsen, K. A., and A. F. Horwitz. 1977. Dev. Biol. 58:328-338) and is mediated by cell surface glycoproteins (Knudsen, K. A., 1985. J. Cell Biol. 101:891- 897). In this paper we show that myoblast adhesion and myotube formation are inhibited by treating fusion-competent myoblasts with phosphatidylinosito...

  1. Establishment of cell surface engineering and its development.

    Science.gov (United States)

    Ueda, Mitsuyoshi

    2016-07-01

    Cell surface display of proteins/peptides has been established based on mechanisms of localizing proteins to the cell surface. In contrast to conventional intracellular and extracellular (secretion) expression systems, this method, generally called an arming technology, is particularly effective when using yeasts as a host, because the control of protein folding that is often required for the preparation of proteins can be natural. This technology can be employed for basic and applied research purposes. In this review, I describe various strategies for the construction of engineered yeasts and provide an outline of the diverse applications of this technology to industrial processes such as the production of biofuels and chemicals, as well as bioremediation and health-related processes. Furthermore, this technology is suitable for novel protein engineering and directed evolution through high-throughput screening, because proteins/peptides displayed on the cell surface can be directly analyzed using intact cells without concentration and purification. Functional proteins/peptides with improved or novel functions can be created using this beneficial, powerful, and promising technique. PMID:27305282

  2. Assessing irrigated agriculture's surface water and groundwater consumption by combining satellite remote sensing and hydrologic modelling.

    Science.gov (United States)

    Peña-Arancibia, Jorge L; Mainuddin, Mohammed; Kirby, John M; Chiew, Francis H S; McVicar, Tim R; Vaze, Jai

    2016-01-15

    Globally, irrigation accounts for more than two thirds of freshwater demand. Recent regional and global assessments indicate that groundwater extraction (GWE) for irrigation has increased more rapidly than surface water extraction (SWE), potentially resulting in groundwater depletion. Irrigated agriculture in semi-arid and arid regions is usually from a combination of stored surface water and groundwater. This paper assesses the usefulness of remotely-sensed (RS) derived information on both irrigation dynamics and rates of actual evapotranspiration which are both input to a river-reach water balance model in order to quantify irrigation water use and water provenance (either surface water or groundwater). The assessment is implemented for the water-years 2004/05-2010/11 in five reaches of the Murray-Darling Basin (Australia); a heavily regulated basin with large irrigated areas and periodic droughts and floods. Irrigated area and water use are identified each water-year (from July to June) through a Random Forest model which uses RS vegetation phenology and actual evapotranspiration as predicting variables. Both irrigated areas and actual evapotranspiration from irrigated areas were compared against published estimates of irrigated areas and total water extraction (SWE+GWE).The river-reach model determines the irrigated area that can be serviced with stored surface water (SWE), and the remainder area (as determined by the Random Forest Model) is assumed to be supplemented by groundwater (GWE). Model results were evaluated against observed SWE and GWE. The modelled SWE generally captures the observed interannual patterns and to some extent the magnitudes, with Pearson's correlation coefficients >0.8 and normalised root-mean-square-errormodelling. The RS irrigated areas and actual evapotranspiration can be used to: (i) understand irrigation dynamics, (ii) constrain irrigation models in data scarce regions, as well as (iii) pinpointing areas that require better ground

  3. Combined effects of surface conditions, boundary layer dynamics and chemistry on diurnal SOA-evolution

    Directory of Open Access Journals (Sweden)

    R. H. H. Janssen

    2012-04-01

    Full Text Available We study the combined effects of land surface conditions, atmospheric boundary layer dynamics and chemistry on the diurnal evolution of biogenic secondary organic aerosol in the atmospheric boundary layer, using a model that contains the essentials of all these components. First, we evaluate the model for a case study in Hyytiälä, Finland, and find that it is able to well reproduce the observed dynamics and gas-phase chemistry. We show that the exchange of organic aerosol between the free troposphere and the boundary layer (entrainment must be taken into account in order to explain the observed diurnal cycle in organic aerosol (OA concentration. An examination of the budgets of organic aerosol and terpene concentration shows that the former is dominated by entrainment, while the latter is mainly driven by emission and chemical transformation. We systematically examine the role of the land surface, which governs both the surface energy balance partitioning and terpene-emissions, and the large-scale atmospheric process of vertical subsidence. Entrainment is especially important for the dilution of organic aerosol concentrations under conditions of dry soils and low terpene-emissions. Subsidence suppresses boundary layer growth while enhancing entrainment. Therefore it influences the relationship between organic aerosol and terpene-concentrations. Our findings indicate that the diurnal evolution of SOA in the boundary layer is the result of coupled effects of the land surface, dynamics of the atmospheric boundary layer, chemistry, and free troposphere conditions. This has potentially some consequences for the design of both field campaigns and large-scale modeling studies.

  4. Combined effects of surface conditions, boundary layer dynamics and chemistry on diurnal SOA evolution

    Directory of Open Access Journals (Sweden)

    R. H. H. Janssen

    2012-08-01

    Full Text Available We study the combined effects of land surface conditions, atmospheric boundary layer dynamics and chemistry on the diurnal evolution of biogenic secondary organic aerosol in the atmospheric boundary layer, using a model that contains the essentials of all these components. First, we evaluate the model for a case study in Hyytiälä, Finland, and find that it is able to satisfactorily reproduce the observed dynamics and gas-phase chemistry. We show that the exchange of organic aerosol between the free troposphere and the boundary layer (entrainment must be taken into account in order to explain the observed diurnal cycle in organic aerosol (OA concentration. An examination of the budgets of organic aerosol and terpene concentrations show that the former is dominated by entrainment, while the latter is mainly driven by emission and chemical transformation. We systematically investigate the role of the land surface, which governs both the surface energy balance partitioning and terpene emissions, and the large-scale atmospheric process of vertical subsidence. Entrainment is especially important for the dilution of organic aerosol concentrations under conditions of dry soils and low terpene emissions. Subsidence suppresses boundary layer growth while enhancing entrainment. Therefore, it influences the relationship between organic aerosol and terpene concentrations. Our findings indicate that the diurnal evolution of secondary organic aerosols (SOA in the boundary layer is the result of coupled effects of the land surface, dynamics of the atmospheric boundary layer, chemistry, and free troposphere conditions. This has potentially some consequences for the design of both field campaigns and large-scale modeling studies.

  5. A practical algorithm for estimating surface soil moisture using combined optical and thermal infrared data

    Science.gov (United States)

    Leng, Pei; Song, Xiaoning; Duan, Si-Bo; Li, Zhao-Liang

    2016-10-01

    Surface soil moisture (SSM) is a critical variable for understanding the energy and water exchange between the land and atmosphere. A multi-linear model was recently developed to determine SSM using ellipse variables, namely, the center horizontal coordinate (x0), center vertical coordinate (y0), semi-major axis (a) and rotation angle (θ), derived from the elliptical relationship between diurnal cycles of land surface temperature (LST) and net surface shortwave radiation (NSSR). However, the multi-linear model has a major disadvantage. The model coefficients are calculated based on simulated data produced by a land surface model simulation that requires sufficient meteorological measurements. This study aims to determine the model coefficients directly using limited meteorological parameters rather than via the complicated simulation process, decreasing the dependence of the model coefficients on meteorological measurements. With the simulated data, a practical algorithm was developed to estimate SSM based on combined optical and thermal infrared data. The results suggest that the proposed approach can be used to determine the coefficients associated with all ellipse variables based on historical meteorological records, whereas the constant term varies daily and can only be determined using the daily maximum solar radiation in a prediction model. Simulated results from three FLUXNET sites over 30 cloud-free days revealed an average root mean square error (RMSE) of 0.042 m3/m3 when historical meteorological records were used to synchronously determine the model coefficients. In addition, estimated SSM values exhibited generally moderate accuracies (coefficient of determination R2 = 0.395, RMSE = 0.061 m3/m3) compared to SSM measurements at the Yucheng Comprehensive Experimental Station.

  6. Assessing irrigated agriculture's surface water and groundwater consumption by combining satellite remote sensing and hydrologic modelling.

    Science.gov (United States)

    Peña-Arancibia, Jorge L; Mainuddin, Mohammed; Kirby, John M; Chiew, Francis H S; McVicar, Tim R; Vaze, Jai

    2016-01-15

    Globally, irrigation accounts for more than two thirds of freshwater demand. Recent regional and global assessments indicate that groundwater extraction (GWE) for irrigation has increased more rapidly than surface water extraction (SWE), potentially resulting in groundwater depletion. Irrigated agriculture in semi-arid and arid regions is usually from a combination of stored surface water and groundwater. This paper assesses the usefulness of remotely-sensed (RS) derived information on both irrigation dynamics and rates of actual evapotranspiration which are both input to a river-reach water balance model in order to quantify irrigation water use and water provenance (either surface water or groundwater). The assessment is implemented for the water-years 2004/05-2010/11 in five reaches of the Murray-Darling Basin (Australia); a heavily regulated basin with large irrigated areas and periodic droughts and floods. Irrigated area and water use are identified each water-year (from July to June) through a Random Forest model which uses RS vegetation phenology and actual evapotranspiration as predicting variables. Both irrigated areas and actual evapotranspiration from irrigated areas were compared against published estimates of irrigated areas and total water extraction (SWE+GWE).The river-reach model determines the irrigated area that can be serviced with stored surface water (SWE), and the remainder area (as determined by the Random Forest Model) is assumed to be supplemented by groundwater (GWE). Model results were evaluated against observed SWE and GWE. The modelled SWE generally captures the observed interannual patterns and to some extent the magnitudes, with Pearson's correlation coefficients >0.8 and normalised root-mean-square-errordata scarce regions, as well as (iii) pinpointing areas that require better ground-based monitoring. PMID:26520262

  7. Surface roughness effect on the metallic bipolar plates of a proton exchange membrane fuel cell

    International Nuclear Information System (INIS)

    Highlights: ► Various degrees of roughness are caused by the sandblasting method. ► An improper surface modification depletes the PEMFC performance severely. ► The AC impedance are used to assess the fuel gas transfer effect. ► The Warburg resistance form in the coarse flow channel surface. - Abstract: Proton exchange membrane fuel cells (PEMFCs) is a promising candidate as energy systems. However, the stability and lifetime of cells are still important issues. The effect of surface roughness on metallic bipolar plate is discussed in this paper. Various roughness on the bulk surface are obtained by the sandblasting method. The grain sizes of sand are selected as 50, 100 and 200 μm. The Ac impedance experiment results show that the bipolar plate roughness and carbon paper porosity are well matched when the surface roughness is within 1–2 μm. Superior condition decreases the contact resistance loss in the fuel cell. The high frequency resistance of the coarse surface was larger than that of the substrate by around 5 mΩ. Furthermore, a new arc was formed at the low frequency region. Hence, the unmatch roughness condition of the bipolar plate significantly increases the contact resistance and mass transfer resistance. This paper develops a sequential approach to study an optimum surface roughness by combining the whole performance (I–V) curve and AC impedance result. It benefits us to quantify the contact and mass transfer resistance exists in the PEMFC. The proposed surface treatment improves the surface effect and promotes the implement of potential metallic bipolar plate in near future

  8. Anomalous cell surface structure of sickle cell anemia erythrocytes as demonstrated by cell surface labeling and endo-beta-galactosidase treatment

    Energy Technology Data Exchange (ETDEWEB)

    Fukuda, M.; Fukuda, M.N.; Hakomori, S.; Papayannopoulou, T.

    1981-01-01

    Erythrocyte surface glycoproteins from patients with various types of sickle cell anemia have been analyzed and compared with those from normal individuals. By hemagglutination with various anti-carbohydrate antibodies, sickle cells showed profound increase of i antigens and moderate increase of GlcNAc beta 1 leads to 3Gal beta 1 leads to 3 Glc structure, whereas antigenicity toward globosidic structure was unchanged. In parallel to these findings, erythrocytes of sickle cell patients have additional sialylated lactosaminoglycan in Band 3. Thus, it can be concluded that erythrocytes of sickle cell patients are characterized by an altered cell surface structure which does not appear to be due to topographical changes of cell surface membrane. It is possible that the anemia or the ''stress'' hematopoiesis in these patients is responsible for these changes.

  9. Enhancement of photovoltaic properties of multicrystalline silicon solar cells by combination of buried metallic contacts and thin porous silicon

    Energy Technology Data Exchange (ETDEWEB)

    Ben Rabha, M.; Bessais, B. [Laboratoire de Photovoltaique, Centre de Recherches et des Technologies de l' Energie, Technopole de Borj-Cedria, BP 95, 2050 Hammam-Lif (Tunisia)

    2010-03-15

    Photovoltaic properties of buried metallic contacts (BMCs) with and without application of a front porous silicon (PS) layer on multicrystalline silicon (mc-Si) solar cells were investigated. A Chemical Vapor Etching (CVE) method was used to perform front PS layer and BMCs of mc-Si solar cells. Good electrical performance for the mc-Si solar cells was observed after combination of BMCs and thin PS films. As a result the current-voltage (I-V) characteristics and the internal quantum efficiency (IQE) were improved, and the effective minority carrier diffusion length (Ln) increases from 75 to 110 {mu}m after BMCs achievement. The reflectivity was reduced to 8% in the 450-950 nm wavelength range. This simple and low cost technology induces a 12% conversion efficiency (surface area = 3.2 cm{sup 2}). The obtained results indicate that the BMCs improve charge carrier collection while the PS layer passivates the front surface. (author)

  10. Brown spider venom toxins interact with cell surface and are endocytosed by rabbit endothelial cells.

    Science.gov (United States)

    Nowatzki, Jenifer; de Sene, Reginaldo Vieira; Paludo, Katia Sabrina; Veiga, Silvio Sanches; Oliver, Constance; Jamur, Maria Célia; Nader, Helena Bonciani; Trindade, Edvaldo S; Franco, Célia Regina C

    2010-09-15

    Bites from the Loxosceles genus (brown spiders) cause severe clinical symptoms, including dermonecrotic injury, hemorrhage, hemolysis, platelet aggregation and renal failure. Histological findings of dermonecrotic lesions in animals exposed to Loxosceles intermedia venom show numerous vascular alterations. Study of the hemorrhagic consequences of the venom in endothelial cells has demonstrated that the degeneration of blood vessels results not only from degradation of the extracellular matrix molecule or massive leukocyte infiltration, but also from a direct and primary activity of the venom on endothelial cells. Exposure of an endothelial cell line in vitro to L. intermedia venom induce morphological alterations, such as cell retraction and disadhesion to the extracellular matrix. The aim of the present study was to investigate the interaction between the venom toxins and the endothelial cell surface and their possible internalization, in order to illuminate the information about the deleterious effect triggered by venom. After treating endothelial cells with venom toxins, we observed that the venom interacts with cell surface. Venom treatment also can cause a reduction of cell surface glycoconjugates. When cells were permeabilized, it was possible to verify that some venom toxins were internalized by the endothelial cells. The venom internalization involves endocytic vesicles and the venom was detected in the lysosomes. However, no damage to lysosomal integrity was observed, suggesting that the cytotoxic effect evoked by L. intermedia venom on endothelial cells is not mediated by venom internalization.

  11. Stable isotope labeling of oligosaccharide cell surface antigens

    Energy Technology Data Exchange (ETDEWEB)

    Unkefer, C.J.; Silks, L.A. III; Martinez, R.A. [and others

    1998-12-31

    The overall goal of this Laboratory Directed Research and Development (LDRD) project was to develop new methods for synthesis of {sup 13}C-labeled oligosaccharides that are required for nuclear magnetic resonance (NMR) studies of their solution conformation. Oligosaccharides are components of the cell`s outer surface and are involved in important processes such as cell-cell recognition and adhesion. Recently, Danishefsky and coworkers at Slone-Kettering Cancer Center developed a method for the solid-phase chemical synthesis of oligosaccharides. The specific goal of this LDRD project was to prepare uniform {sup 13}C-labeled aldohexose precursors required for the solid-phase synthesis of the Lewis blood-group antigenic determinants. We report the synthesis of {sup 13}C-labeled D-glucal, D-galactal and Fucosyl precursors. We have been collaborating with the Danishefsky group on the synthesis of the Lewis oligosaccharides and the NMR analysis of their solution conformation.

  12. Combined Papillated Bowen Disease and Clear Cell Atypical Fibroxanthoma

    Directory of Open Access Journals (Sweden)

    Dimas Suárez-Vilela

    2010-05-01

    Full Text Available We describe a case of papillated Bowen disease (PBD, associated with a clear cell atypical fibroxanthoma (CCAFXA. The epidermal lesion showed a bowenoid papillomatous growth pattern with histologic features suggestive of infection by human papilloma virus (HPV. In the dermis a neoplasm made up by spindled or polygonal cells with wide clear cytoplasm and moderate nuclear pleomorphism was found. Immunohistochemical characteristics of these two lesions were clearly different. The atypical cells of the intraepidermal proliferation were positive for AE1-AE3 anticytokeratin antibody, EMA, p16, p53 and p63. The dermal tumor was positive for vimentin, CD10, CD68, CD99, alpha-1-antitrypsin and c-kit. Histological features and immunohistochemical profile of the dermal tumor corresponded to a CCAFXA, a very uncommon neoplasm of which only 10 cases have been reported. In situ hybridization for numerous types of HPVs was negative in both lesions.

  13. Elucidating the signal responses of multi-parametric surface plasmon resonance living cell sensing: a comparison between optical modeling and drug-MDCKII cell interaction measurements.

    Directory of Open Access Journals (Sweden)

    Tapani Viitala

    Full Text Available In vitro cell-based assays are widely used during the drug discovery and development process to test the biological activity of new drugs. Most of the commonly used cell-based assays, however, lack the ability to measure in real-time or under dynamic conditions (e.g. constant flow. In this study a multi-parameter surface plasmon resonance approach in combination with living cell sensing has been utilized for monitoring drug-cell interactions in real-time, under constant flow and without labels. The multi-parameter surface plasmon resonance approach, i.e. surface plasmon resonance angle versus intensity plots, provided fully specific signal patterns for various cell behaviors when stimulating cells with drugs that use para- and transcellular absorption routes. Simulated full surface plasmon resonance angular spectra of cell monolayers were compared with actual surface plasmon resonance measurements performed with MDCKII cell monolayers in order to better understand the origin of the surface plasmon resonance signal responses during drug stimulation of cells. The comparison of the simulated and measured surface plasmon resonance responses allowed to better understand and provide plausible explanations for the type of cellular changes, e.g. morphological or mass redistribution in cells, that were induced in the MDCKII cell monolayers during drug stimulation, and consequently to differentiate between the type and modes of drug actions. The multi-parameter surface plasmon resonance approach presented in this study lays the foundation for developing new types of cell-based tools for life science research, which should contribute to an improved mechanistic understanding of the type and contribution of different drug transport routes on drug absorption.

  14. Accelerated differentiation of osteoblast cells on polycaprolactone scaffolds driven by a combined effect of protein coating and plasma modification

    Energy Technology Data Exchange (ETDEWEB)

    Yildirim, Eda D; Gueceri, Selcuk; Sun, Wei [Department of Mechanical Engineering and Mechanics, Drexel University, 3141 Chestnut Street, Philadelphia, PA 19104 (United States); Besunder, Robyn; Allen, Fred [Drexel University, School of Biomedical Engineering Science and Health System, 3141 Chestnut Street, Philadelphia, PA 19104 (United States); Pappas, Daphne, E-mail: edy22@drexel.ed [Army Research Laboratory, Aberdeen Proving Ground, MD 21005 (United States)

    2010-03-15

    A combined effect of protein coating and plasma modification on the quality of the osteoblast-scaffold interaction was investigated. Three-dimensional polycaprolactone (PCL) scaffolds were manufactured by the precision extrusion deposition (PED) system. The structural, physical, chemical and biological cues were introduced to the surface through providing 3D structure, coating with adhesive protein fibronectin and modifying the surface with oxygen-based plasma. The changes in the surface properties of PCL after those modifications were examined by contact angle goniometry, surface energy calculation, surface chemistry analysis (XPS) and surface topography measurements (AFM). The effects of modification techniques on osteoblast short-term and long-term functions were examined by cell adhesion, proliferation assays and differentiation markers, namely alkaline phosphatase activity (ALP) and osteocalcin secretion. The results suggested that the physical and chemical cues introduced by plasma modification might be sufficient for improved cell adhesion, but for accelerated osteoblast differentiation the synergetic effects of structural, physical, chemical and biological cues should be introduced to the PCL surface.

  15. Osteocytic cell necrosis is caused by a combination of glucocorticoid-induced Dickkopf-1 and hypoxia.

    Science.gov (United States)

    Ueda, Shusuke; Ichiseki, Toru; Yoshitomi, Yasuo; Yonekura, Hideto; Ueda, Yoshimichi; Kaneuji, Ayumi; Matsumoto, Tadami

    2015-06-01

    Osteonecrosis is a major glucocorticoid-induced complication in the orthopedics field. Despite the extensive researches, mechanisms underlining the glucocorticoid-induced osteonecrosis are largely unknown. Here, we first provide the evidence that a combined treatment of cultured osteocytic cells with glucocorticoid and hypoxia caused necrotic cell death, which is assumed to occur in the acute bone injuries induced by glucocorticoids. We cultured MLO-Y4 murine osteocytic cells under hypoxia in the presence or absence of Dexamethasone (Dex) and examined the rates of apoptotic and necrotic cell death. Dex or hypoxia alone increased apoptotic cells, but not necrotic cells. The combination of Dex and hypoxia dramatically increased osteocytic cell death, notably necrotic cell death. The expression of Dickkopf-1 (Dkk-1), an inhibitor of Wnt/β-catenin signal, was scarcely expressed in the control and hypoxic cells, but a dramatic increase of the Dkk-1 expression was detected in Dex-treated cells. siRNA-mediated knockdown of Dkk-1 in Dex and hypoxia-treated osteocytic cells showed the significant decreases in both apoptotic and necrotic cells. The results indicated that the combination of Dkk-1 overexpression by Dex and hypoxia causes the necrotic osteocytic cell death. The results also indicated that blocking of Dkk-1 can protect bone cells from glucocorticoid and hypoxia-induced cell injury. PMID:24819581

  16. Bacterial Cell Surface Adsorption of Rare Earth Elements

    Science.gov (United States)

    Jiao, Y.; Park, D.; Reed, D.; Fujita, Y.; Yung, M.; Anderko, A.; Eslamimanesh, A.

    2015-12-01

    Rare earth elements (REE) play a critical role in many emerging clean energy technologies, including high-power magnets, wind turbines, solar panels, hybrid/electric vehicle batteries and lamp phosphors. In order to sustain demand for such technologies given current domestic REE shortages, there is a need to develop new approaches for ore processing/refining and recycling of REE-containing materials. To this end, we have developed a microbially-mediated bioadsorption strategy with application towards enrichment of REE from complex mixtures. Specifically, the bacterium Caulobacter crescentus was genetically engineered to display lanthanide binding tags (LBTs), short peptides that possess high affinity and specificity for rare earth elements, on its cell surface S-layer protein. Under optimal conditions, LBT-displayed cells adsorbed greater than 5-fold more REE than control cells lacking LBTs. Competition binding experiments with a selection of REEs demonstrated that our engineered cells could facilitate separation of light- from heavy- REE. Importantly, binding of REE onto our engineered strains was much more favorable compared to non-REE metals. Finally, REE bound to the cell surface could be stripped off using citrate, providing an effective and non-toxic REE recovery method. Together, this data highlights the potential of our approach for selective REE enrichment from REE containing mixtures.

  17. Methodology for optimizing the electropolishing of stainless steel AISI 316L combining criteria of surface finish and dimensional precision

    Science.gov (United States)

    Núñez, P. J.; García-Plaza, E.; Martín, A. R.; Trujillo, R.; De la Cruz, C.

    2009-11-01

    This work examines a methodology for optimizing electrochemical polishing conditions bearing in mind the criteria that enhance minimum surface roughness and dimensional precision (minimum loss of thickness). The study consisted in electrochemically polishing stainless steel AISI 316L (ISO 4954 X2CrNiMo17133E) under a combination of different temperatures (T) baths and current densities (J), and application times (t). The surface finish (ΔRa) and dimensional variations (Δh) of the electrochemically polished workpieces were assessed, and the experimental data of the variables was correlated as can be seen by the response surfaces. This methodology enables optimum working areas to be specified using the sole criteria of surface finish, or by using a combination of both criteria (minimum roughness and maximum precision). The methodology has proven to be an optimum method for selecting electrochemical polishing conditions using the combined criteria of surface finish and dimensional precision in accordance with design requirements.

  18. Surface water quality assessment by the use of combination of multivariate statistical classification and expert information.

    Science.gov (United States)

    Tobiszewski, M; Tsakovski, S; Simeonov, V; Namieśnik, J

    2010-08-01

    The present study deals with the assessment of surface water quality from an industrial-urban region located in northern Poland near to the city of Gdansk. Concentrations of thirteen chemicals including total polycyclic aromatic hydrocarbons (PAHs), halogenated volatile organic compounds (HVOCs) and major ions in the samples collected at five sampling points during six campaigns were used as variables throughout the study. The originality in the monitoring data treatment and interpretation was the combination of a traditional classification approach (self-organizing maps of Kohonen) with PAH diagnostic ratios expertise to achieve a reliable pollution source identification. Thus, sampling points affected by pollution from traffic (petroleum combustion products), from crude oil processing (petroleum release related compounds), and from phosphogypsum disposal site were properly discriminated. Additionally, it is shown that this original assessment approach can be useful in finding specific pollution source tracers.

  19. Plasma formation on a metal surface under combined action of laser and microwave radiation

    Energy Technology Data Exchange (ETDEWEB)

    Gavrilyuk, A P; Shaparev, N Ya [Institute of Computational Modelling of Siberian Branch of the Russian Academy of Sciences, Krasnoyarsk (Russian Federation)

    2013-10-31

    By means of numerical modelling of the combined effect of laser (1.06 mm) and microwave (10{sup 10} – 10{sup 13} s{sup -1}) radiation on the aluminium surface in vacuum it is shown that the additional action of microwave radiation with the frequency 10{sup 12} s{sup -1} provides complete ionisation of the metal vapour (for the values of laser radiation duration and intensity used in the calculations), while in the absence of microwave radiation the vapour remains weakly ionised. The mathematical model used accounts for the processes, occurring in the condensed phase (heat conduction, melting), the evaporation and the kinetic processes in the resulting vapour. (interaction of laser radiation with matter. laser plasma)

  20. Mapping surface energy balance components by combining landsat thematic mapper and ground-based meteorological data

    Science.gov (United States)

    Moran, M.S.; Jackson, R. D.; Raymond, L.H.; Gay, L.W.; Slater, P.N.

    1989-01-01

    Surface energy balance components were evaluated by combining satellite-based spectral data with on-site measurements of solar irradiance, air temperature, wind speed, and vapor pressure. Maps of latent heat flux density (??E) and net radiant flux density (Rn) were produced using Landsat Thematic Mapper (TM) data for three dates: 23 July 1985, 5 April 1986, and 24 June 1986. On each date, a Bowen-ratio apparatus, located in a vegetated field, was used to measure ??E and Rn at a point within the field. Estimates of ??E and Rn were also obtained using radiometers aboard an aircraft flown at 150 m above ground level. The TM-based estimates differed from the Bowen-ratio and aircraft-based estimates by less than 12 % over mature fields of cotton, wheat, and alfalfa, where ??E and Rn ranged from 400 to 700 Wm-2. ?? 1989.

  1. Eliminating Cancer Stem Cells in CML with Combination Transcriptional Therapy.

    Science.gov (United States)

    Carvajal, Luis A; Steidl, Ulrich

    2016-07-01

    Leukemia stem cells (LSCs) are resistant to current therapies used to treat chronic myeloid leukemia (CML). Abraham et al. (2016) have identified a molecular network critical for CML LSC survival and propose that simultaneously targeting two of their major transcriptional regulators, p53 and c-Myc, may facilitate their eradication. PMID:27392220

  2. Optical tweezers and surface plasmon resonance combination system based on the high numerical aperture lens

    Science.gov (United States)

    Shan, Xuchen; Zhang, Bei; Lan, Guoqiang; Wang, Yiqiao; Liu, Shugang

    2015-11-01

    Biology and medicine sample measurement takes an important role in the microscopic optical technology. Optical tweezer has the advantage of accurate capture and non-pollution of the sample. The SPR(surface plasmon resonance) sensor has so many advantages include high sensitivity, fast measurement, less consumption of sample and label-free detection of biological sample that the SPR sensing technique has been used for surface topography, analysis of biochemical and immune, drug screening and environmental monitoring. If they combine, they will play an important role in the biological, chemical and other subjects. The system we propose use the multi-axis cage system, by using the methods of reflection and transmiss ion to improve the space utilization. The SPR system and optical tweezer were builtup and combined in one system. The cage of multi-axis system gives full play to its accuracy, simplicity and flexibility. The size of the system is 20 * 15 * 40 cm3 and thus the sample can be replaced to switch between the optical tweezers system and the SPR system in the small space. It means that we get the refractive index of the sample and control the particle in the same system. In order to control the revolving stage, get the picture and achieve the data stored automatically, we write a LabVIEW procedure. Then according to the data from the back focal plane calculate the refractive index of the sample. By changing the slide we can trap the particle as optical tweezer, which makes us measurement and trap the sample at the same time.

  3. Vaccines based on the cell surface carbohydrates of pathogenic bacteria

    Directory of Open Access Journals (Sweden)

    Jones Christopher

    2005-01-01

    Full Text Available Glycoconjugate vaccines, in which a cell surface carbohydrate from a micro-organism is covalently attached to an appropriate carrier protein are proving to be the most effective means to generate protective immune responses to prevent a wide range of diseases. The technology appears to be generic and applicable to a wide range of pathogens, as long as antibodies against surface carbohydrates help protect against infection. Three such vaccines, against Haemophilus influenzae type b, Neisseria meningitidis Group C and seven serotypes of Streptococcus pneumoniae, have already been licensed and many others are in development. This article discusses the rationale for the development and use of glycoconjugate vaccines, the mechanisms by which they elicit T cell-dependent immune responses and the implications of this for vaccine development, the role of physicochemical methods in the characterisation and quality control of these vaccines, and the novel products which are under development.

  4. Active screen plasma nitriding enhances cell attachment to polymer surfaces

    Energy Technology Data Exchange (ETDEWEB)

    Kaklamani, Georgia, E-mail: g.kaklamani@bham.ac.uk [University of Birmingham, College of Engineering and Physical Sciences, School of Metallurgy and Materials, Edgbaston, Birmingham B15 2TT (United Kingdom); Bowen, James; Mehrban, Nazia [University of Birmingham, College of Engineering and Physical Sciences, School of Chemical Engineering, Edgbaston, Birmingham B15 2TT (United Kingdom); Dong, Hanshan [University of Birmingham, College of Engineering and Physical Sciences, School of Metallurgy and Materials, Edgbaston, Birmingham B15 2TT (United Kingdom); Grover, Liam M. [University of Birmingham, College of Engineering and Physical Sciences, School of Chemical Engineering, Edgbaston, Birmingham B15 2TT (United Kingdom); Stamboulis, Artemis [University of Birmingham, College of Engineering and Physical Sciences, School of Metallurgy and Materials, Edgbaston, Birmingham B15 2TT (United Kingdom)

    2013-05-15

    Active screen plasma nitriding (ASPN) is a well-established technique used for the surface modification of materials, the result of which is often a product with enhanced functional performance. Here we report the modification of the chemical and mechanical properties of ultra-high molecular weight poly(ethylene) (UHMWPE) using 80:20 (v/v) N{sub 2}/H{sub 2} ASPN, followed by growth of 3T3 fibroblasts on the treated and untreated polymer surfaces. ASPN-treated UHMWPE showed extensive fibroblast attachment within 3 h of seeding, whereas fibroblasts did not successfully attach to untreated UHMWPE. Fibroblast-coated surfaces were maintained for up to 28 days, monitoring their metabolic activity and morphology throughout. The chemical properties of the ASPN-treated UHMWPE surface were studied using X-ray photoelectron spectroscopy, revealing the presence of C-N, C=N, and C≡N chemical bonds. The elastic modulus, surface topography, and adhesion properties of the ASPN-treated UHMWPE surface were studied over 28 days during sample storage under ambient conditions and during immersion in two commonly used cell culture media.

  5. Combined sequencing of mRNA and DNA from human embryonic stem cells.

    Science.gov (United States)

    Mertes, Florian; Kuhl, Heiner; Wruck, Wasco; Lehrach, Hans; Adjaye, James

    2016-06-01

    Combined transcriptome and whole genome sequencing of the same ultra-low input sample down to single cells is a rapidly evolving approach for the analysis of rare cells. Besides stem cells, rare cells originating from tissues like tumor or biopsies, circulating tumor cells and cells from early embryonic development are under investigation. Herein we describe a universal method applicable for the analysis of minute amounts of sample material (150 to 200 cells) derived from sub-colony structures from human embryonic stem cells. The protocol comprises the combined isolation and separate amplification of poly(A) mRNA and whole genome DNA followed by next generation sequencing. Here we present a detailed description of the method developed and an overview of the results obtained for RNA and whole genome sequencing of human embryonic stem cells, sequencing data is available in the Gene Expression Omnibus (GEO) database under accession number GSE69471.

  6. Extraction of cell surface-associated proteins from living yeast cells.

    NARCIS (Netherlands)

    F.M. Klis; M. de Jong; S. Brul; P.W.J. de Groot

    2007-01-01

    To extract cell surface-associated proteins from living fungal cells, reducing agents such as beta-mercaptoethanol and dithiothreitol are often used. We show here that both compounds are moderately lipophilic and may perturb the plasma membrane, thus causing the release of cytosolic proteins, especi

  7. A Simple Hydrophilic Treatment of SU-8 Surfaces for Cell Culturing and Cell Patterning

    DEFF Research Database (Denmark)

    Wang, Zhenyu; Stangegaard, Michael; Dufva, Hans Martin;

    2005-01-01

    SU-8, an epoxy-based photoresist, widely used in constitution different mTAS systems, is incompatible with mammalian cell adhesion and culture in its native form. Here, we demonstrate a simple, cheap and robust two-step method to render a SU-8 surface hydrophilic and compatible with cell culture...

  8. Methods To Identify Aptamers against Cell Surface Biomarkers

    OpenAIRE

    Frédéric Ducongé; Daniel Miotto Dupont; Agnes Cibiel

    2011-01-01

    Aptamers are nucleic acid-based ligands identified through a process of molecular evolution named SELEX (Systematic Evolution of Ligands by Exponential enrichment). During the last 10-15 years, numerous aptamers have been developed specifically against targets present on or associated with the surface of human cells or infectious pathogens such as viruses, bacteria, fungi or parasites. Several of the aptamers have been described as potent probes, rivalling antibodies, for use in flow cytometr...

  9. Biointerface: protein enhanced stem cells binding to implant surface.

    Science.gov (United States)

    Chrzanowski, W; Kondyurin, A; Lee, Jae Ho; Lord, Megan S; Bilek, M M M; Kim, Hae-Won

    2012-09-01

    The number of metallic implantable devices placed every year is estimated at 3.7 million. This number has been steadily increasing over last decades at a rate of around 8 %. In spite of the many successes of the devices the implantation of biomaterial into tissues almost universally leads to the development of an avascular sac, which consists of fibrous tissue around the device and walls off the implant from the body. This reaction can be detrimental to the function of implant, reduces its lifetime, and necessitates repeated surgery. Clearly, to reduce the number of revision surgeries and improve long-term implant function it is necessary to enhance device integration by modulating cell adhesion and function. In this paper we have demonstrated that it is possible to enhance stem cell attachment using engineered biointerfaces. To create this functional interface, samples were coated with polymer (as a precursor) and then ion implanted to create a reactive interface that aids the binding of biomolecules--fibronectin. Both AFM and XPS analyses confirmed the presence of protein layers on the samples. The amount of protein was significant greater for the ion implanted surfaces and was not disrupted upon washing with detergent, hence the formation of strong bonds with the interface was confirmed. While, for non ion implanted surfaces, a decrease of protein was observed after washing with detergent. Finally, the number of stem cells attached to the surface was enhanced for ion implanted surfaces. The studies presented confirm that the developed bionterface with immobilised fibronectin is an effective means to modulate stem cell attachment. PMID:22714559

  10. Recent Insights into Cell Surface Heparan Sulphate Proteoglycans and Cancer

    DEFF Research Database (Denmark)

    Couchman, John R; Multhaupt, Hinke; Sanderson, Ralph D

    2016-01-01

    A small group of cell surface receptors are proteoglycans, possessing a core protein with one or more covalently attached glycosaminoglycan chains. They are virtually ubiquitous and their chains are major sites at which protein ligands of many types interact. These proteoglycans can signal and re...... or fragmented proteoglycans into exosomes that can be paracrine effectors or biomarkers, and lateral interactions between some proteoglycans and calcium channels that impact the actin cytoskeleton....

  11. Smooth Muscle Cell Functionality on Collagen Immobilized Polycaprolactone Nanowire Surfaces

    OpenAIRE

    Victoria Leszczak; Baskett, Dominique A.; Popat, Ketul C.

    2014-01-01

    Inhibition of smooth muscle cell (SMC) proliferation and preservation of a differentiated state are important aspects in the management, avoidance and progression of vascular diseases. An understanding of the interaction between SMCs and the biomaterial involved is essential for a successful implant. In this study, we have developed collagen immobilized nanostructured surfaces with controlled arrays of high aspect ratio nanowires for the growth and maintenance of human aortic SMCs. The nanow...

  12. Cadmium and lead interaction with diatom surfaces: A combined thermodynamic and kinetic approach

    Science.gov (United States)

    Gélabert, A.; Pokrovsky, O. S.; Schott, J.; Boudou, A.; Feurtet-Mazel, A.

    2007-08-01

    This work is devoted to the physico-chemical study of cadmium and lead interaction with diatom-water interfaces for two marine planktonic ( Thalassiosira weissflogii, TW; Skeletonema costatum, SC) and two freshwater periphytic species ( Achnanthidium minutissimum, AMIN; Navicula minima, NMIN) by combining adsorption measurements with surface complexation modeling. Adsorption kinetics was studied as a function of pH and initial metal concentration in sodium nitrate solution and in culture media. Kinetic data were consistent with a two-step mechanism in which the loss of a water molecule from the inner coordination sphere of the metal is rate limiting. Reversible adsorption experiments, with 3 h of exposure to metal, were performed as a function of pH (2-9), metal concentration in solution (10 -9-10 -3 M), and ionic strength (10 -3-1.0 M). While the shape of pH-dependent adsorption edge is similar among all four diatom species, the constant-pH adsorption isotherm and maximal binding capacities differ. Measurements of electrophoretic mobilities ( μ) revealed negative surface potential for AMIN diatom, however, the absolute value of μ decreases with increase of [Pb 2+] aq suggesting the metal adsorption on negative surface sites. These observations allowed us to construct a surface complexation model (SCM) for cadmium and lead binding by diatom surfaces that postulates the Constant Capacitance of the electric double layer and considers Cd and Pb complexation with mainly carboxylic and, partially, silanol groups. In the full range of investigated Cd concentration, the SCM is able to describe the concentration of adsorbed metal as a function of [Cd 2+] aq without implying the presence of high affinity, low abundance sites, that are typically used to model the metal interactions with natural multi-component organic substances. At the same time, Cd fast initial reaction requires the presence of "highly reactive sites" those concentration represents only 2.5-3% of the

  13. Compensation strategy for machining optical freeform surfaces by the combined on- and off-machine measurement.

    Science.gov (United States)

    Zhang, Xiaodong; Zeng, Zhen; Liu, Xianlei; Fang, Fengzhou

    2015-09-21

    Freeform surface is promising to be the next generation optics, however it needs high form accuracy for excellent performance. The closed-loop of fabrication-measurement-compensation is necessary for the improvement of the form accuracy. It is difficult to do an off-machine measurement during the freeform machining because the remounting inaccuracy can result in significant form deviations. On the other side, on-machine measurement may hides the systematic errors of the machine because the measuring device is placed in situ on the machine. This study proposes a new compensation strategy based on the combination of on-machine and off-machine measurement. The freeform surface is measured in off-machine mode with nanometric accuracy, and the on-machine probe achieves accurate relative position between the workpiece and machine after remounting. The compensation cutting path is generated according to the calculated relative position and shape errors to avoid employing extra manual adjustment or highly accurate reference-feature fixture. Experimental results verified the effectiveness of the proposed method.

  14. Meter-Scale 3-D Models of the Martian Surface from Combining MOC and MOLA Data

    Science.gov (United States)

    Soderblom, Laurence A.; Kirk, Randolph L.

    2003-01-01

    We have extended our previous efforts to derive through controlled photoclinometry, accurate, calibrated, high-resolution topographic models of the martian surface. The process involves combining MGS MOLA topographic profiles and MGS MOC Narrow Angle images. The earlier work utilized, along with a particular MOC NA image, the MOLA topographic profile that was acquired simultaneously, in order to derive photometric and scattering properties of the surface and atmosphere so as to force the low spatial frequencies of a one-dimensional MOC photoclinometric model to match the MOLA profile. Both that work and the new results reported here depend heavily on successful efforts to: 1) refine the radiometric calibration of MOC NA; 2) register the MOC to MOLA coordinate systems and refine the pointing; and 3) provide the ability to project into a common coordinate system, simultaneously acquired MOC and MOLA with a single set of SPICE kernels utilizing the USGS ISIS cartographic image processing tools. The approach described in this paper extends the MOC-MOLA integration and cross-calibration procedures from one-dimensional profiles to full two-dimensional photoclinometry and image simulations. Included are methods to account for low-frequency albedo variations within the scene.

  15. Combination of NK Cells and Cetuximab to Enhance Anti-Tumor Responses in RAS Mutant Metastatic Colorectal Cancer.

    Directory of Open Access Journals (Sweden)

    John Pradeep Veluchamy

    Full Text Available The ability of Natural Killer (NK cells to kill tumor targets has been extensively studied in various hematological malignancies. However, NK cell therapy directed against solid tumors is still in early development. Epidermal Growth Factor Receptor (EGFR targeted therapies using monoclonal antibodies (mAbs such as cetuximab and panitumumab are widely used for the treatment of metastatic colorectal cancer (mCRC. Still, the clinical efficacy of this treatment is hampered by mutations in RAS gene, allowing tumors to escape from anti-EGFR mAb therapy. It is well established that NK cells kill tumor cells by natural cytotoxicity and can in addition be activated upon binding of IgG1 mAbs through Fc receptors (CD16/FcγRIIIa on their surface, thereby mediating antibody dependent cellular cytotoxicity (ADCC. In the current study, activated Peripheral Blood NK cells (PBNK were combined with anti-EGFR mAbs to study their effect on the killing of EGFR+/- cancer cell lines, including those with RAS mutations. In vitro cytotoxicity experiments using colon cancer primary tumors and cell lines COLO320, Caco-2, SW620, SW480 and HT-29, demonstrated that PBNK cells are cytotoxic for a range of tumor cells, regardless of EGFR, RAS or BRAF status and at low E:T ratios. Cetuximab enhanced the cytotoxic activity of NK cells on EGFR+ tumor cells (either RASwt, RASmut or BRAFmut in a CD16 dependent manner, whereas it could not increase the killing of EGFR- COLO320. Our study provides a rationale to strengthen NK cell immunotherapy through a combination with cetuximab for RAS and BRAF mutant mCRC patients.

  16. A combined capillary cooling system for cooling fuel cells

    Energy Technology Data Exchange (ETDEWEB)

    Silva, Ana Paula; Pelizza, Pablo Rodrigo; Galante, Renan Manozzo; Bazzo, Edson [Universidade Federal de Santa Catarina (LabCET/UFSC), Florianopolis, SC (Brazil). Dept. de Engenharia Mecanica. Lab. de Combustao e Engenharia de Sistemas Termicos], Emails: ana@labcet.ufsc.br, pablo@labcet.ufsc.br, renan@labcet.ufsc.br, ebazzo@emc.ufsc.br

    2010-07-01

    The operation temperature control has an important influence over the PEMFC (Proton Exchange Membrane Fuel Cell) performance. A two-phase heat transfer system is proposed as an alternative for cooling and thermal control of PEMFC. The proposed system consists of a CPL (Capillary Pumped Loop) connected to a set of constant conductance heat pipes. In this work ceramic wick and stainless mesh wicks have been used as capillary structure of the CPL and heat pipes, respectively. Acetone has been used as the working fluid for CPL and deionized water for the heat pipes. Experimental results of three 1/4 inch stainless steel outlet diameter heats pipes and one CPL have been carried out and presented in this paper. Further experiments are planned coupling the proposed cooling system to a module which simulates the fuel cell. (author)

  17. Selective induction of oxidative stress in cancer cells via synergistic combinations of agents targeting redox homeostasis.

    Science.gov (United States)

    Akladios, Fady N; Andrew, Scott D; Parkinson, Christopher J

    2015-07-01

    Cancer cell resistance to chemotherapy is still a heavy burden that impairs the response of many cancer patients to conventional chemotherapy. Using drug combinations is one therapeutic approach to overcome the developing resistance to any one drug. Oxidative stress is now a generally regarded hallmark of cancer that can be one approach to selectively target cancer cells while sparing normal cells. With the aim of increasing oxidative stress in cancer cells to a lethal set point, we have generated and combined several series of redox active compounds that act at different points of the cellular oxidative cascade. The premise of such combinations is to deplete of endogenous antioxidant defence proteins (e.g., Glutathione) while concomitantly increasing the generation of ROS via metal redox recycling and Fenton chemistry which eventually leads to the disruption of cellular redox homeostasis and induction of cell death. Through this approach, we have identified highly synergistic combinations of two distinctive classes of compounds (Azines and Copper(II) complexes of 2-pyridyl ketone thiosemicarbazones) which are capable of eliminating cancer cells without concomitant increase in toxicity toward normal cells. In one of our most potent combinations, a combination index (CI) value of 0.056 was observed, representing a 17 fold enhancement in activity beyond additive effects. Such new combination regimen of redox active compounds can be one step closer to potentially safer low dose chemotherapy. PMID:26022081

  18. Comparing and Combining Remotely Sensed Land Surface Temperature Products for Improved Hydrological Applications

    Directory of Open Access Journals (Sweden)

    Robert M. Parinussa

    2016-02-01

    Full Text Available Land surface temperature (LST is an important variable that provides a valuable connection between the energy and water budget and is strongly linked to land surface hydrology. Space-borne remote sensing provides a consistent means for regularly observing LST using thermal infrared (TIR and passive microwave observations each with unique strengths and weaknesses. The spatial resolution of TIR based LST observations is around 1 km, a major advantage when compared to passive microwave observations (around 10 km. However, a major advantage of passive microwaves is their cloud penetrating capability making them all-weather sensors whereas TIR observations are routinely masked under the presence of clouds and aerosols. In this study, a relatively simple combination approach that benefits from the cloud penetrating capacity of passive microwave sensors was proposed. In the first step, TIR and passive microwave LST products were compared over Australia for both anomalies and raw timeseries. A very high agreement was shown over the vast majority of the country with R2 typically ranging from 0.50 to 0.75 for the anomalies and from 0.80 to 1.00 for the raw timeseries. Then, the scalability of the passive microwave based LST product was examined and a pixel based merging approach through linear scaling was proposed. The individual and merged LST products were further compared against independent LST from the re-analysis model outputs. This comparison revealed that the TIR based LST product agrees best with the re-analysis data (R2 0.26 for anomalies and R2 0.76 for raw data, followed by the passive microwave LST product (R2 0.16 for anomalies and R2 0.66 for raw data and the combined LST product (R2 0.18 for anomalies and R2 0.62 for raw data. It should be noted that the drop in performance comes with an increased revisit frequency of approximately 20% compared to the revised frequency of the TIR alone. Additionally, this comparison against re

  19. Combining a 2-D multiphase CFD model with a Response Surface Methodology to optimize the gasification of Portuguese biomasses

    International Nuclear Information System (INIS)

    Highlights: • A multiphase CFD model was combined with RSM. • Gasification optimal operating conditions were found in a pilot scale reactor. • Syngas quality indices were optimized in a biomass gasification process. • Propagation of error methodology was combined with a CFD model and RSM. - Abstract: This paper presents a study to evaluate the potential of Portuguese biomasses (coffee husks, forest residues and vine pruning residues) to produce syngas for different applications. By using a 2-D Eulerian–Eulerian approach within the CFD framework, a design of several computer experiments was developed and were used as analysis tools the response surface method (RSM) and the propagation of error (POE) approach. The CFD model was validated under experimental results collected at a semi-industrial reactor. For design purposes, temperature, steam to biomass ratio (SBR) and the type of biomass were selected as input factors. The responses were the H2 generation, the H2/CO ratio, the CH4/H2 ratio, the carbon conversion and the cold gas efficiency. It was concluded that after an optimization procedure to determine the operating conditions, vine pruning residues could show very promising results considering some of the typical syngas indice standards for commercial purposes. From the optimization procedure, it was also concluded that forest residues are preferable for domestic natural gas applications and vine pruning residues for fuel cells and integrated gasification systems application. By using the RSM combined with POE, it was verified that the operating conditions to get higher performances do not always coincide with those necessary to obtain a stable syngas composition

  20. Ocular surface evaluation in patients treated with a fixed combination of prostaglandin analogues with 0.5% timolol maleate topical monotherapy: a randomized clinical trial

    Directory of Open Access Journals (Sweden)

    Heloisa Helena Russ

    2013-10-01

    Full Text Available OBJECTIVES: To compare ocular surface changes induced via glaucoma treatment in patients using fixed combinations of prostaglandin analogues (travoprost, latanoprost and bimatoprost with 0.5% timolol maleate METHODS: A prospective, multicenter, randomized, parallel group, single-blind clinical trial was performed in 33 patients with ocular hypertension or open angle glaucoma who had not been previously treated. The ocular surface was evaluated prior to and three months after treatment, with a daily drop instillation of one of the three medications. The main outcome measurements included the tear film break-up time, Schirmer's test, Lissamine green staining, the Ocular Surface Disease Index questionnaire, impression cytology using HE and PAS and immunocytochemistry for interleukin-6 and HLA-DR. Ensaiosclinicos.gov.br: UTN - U1111-1129-2872 RESULTS: All of the drugs induced a significant reduction in intraocular pressure. Decreases in the Schirmer's test results were observed with all of the drugs. Decreases in tear-film break-up time were noted with travoprost/timolol and latanoprost/timolol. An increase in the Lissamine green score was noted with travoprost/timolol and bimatoprost/timolol. The Ocular Surface Disease Index score increased after treatment in the travoprost/timolol group. Impression cytology revealed a significant difference in cell-to-cell contact in the same group, an increase in cellularity in all of the groups and an increase in the number of goblet cells in all of the groups. The fixed combinations induced an increase in IL-6 expression in the travoprost/timolol group, in which there was also an increase in HLA-DR expression. CONCLUSIONS: All of the fixed combinations induced a significant reduction in intraocular pressure, and the travoprost/timolol group showed increased expression of the inflammatory markers HLA-DR and interleukin-6. All three tested medications resulted in some degree of deterioration in the ocular surface

  1. Cell-surface acceleration of urokinase-catalyzed receptor cleavage

    DEFF Research Database (Denmark)

    Høyer-Hansen, G; Ploug, M; Behrendt, N;

    1997-01-01

    The urokinase-type plasminogen activator (uPA) binds to a specific cell-surface receptor, uPAR. On several cell types uPAR is present both in the full-length form and a cleaved form, uPAR(2+3), which is devoid of binding activity. The formation of uPAR(2+3) on cultured U937 cells is either directly...... by a prior incubation of the cells with uPA inactivated by diisopropyl fluorophosphate, demonstrating a requirement for specific receptor binding of the active uPA to obtain the high-efficiency cleavage of cell-bound uPAR. Furthermore, amino-terminal sequence analysis revealed that uPAR(2+3), purified from U......937 cell lysates, had the same amino termini as uPAR(2+3), generated by uPA in a purified system. In both cases cleavage had occurred at two positions in the hinge region connecting domain 1 and 2, between Arg83-Ala84 and Arg89-Ser90, respectively. The uPA-catalyzed cleavage of uPAR is a new negative...

  2. Advances in targeting cell surface signalling molecules for immune modulation

    Science.gov (United States)

    Yao, Sheng; Zhu, Yuwen; Chen, Lieping

    2013-01-01

    The past decade has witnessed a surge in the development of immunomodulatory approaches to combat a broad range of human diseases, including cancer, viral infections, autoimmunity and inflammation as well as in the prevention of transplant rejection. Immunomodulatory approaches mostly involve the use of monoclonal antibodies or recombinant fusion proteins that target cell surface signalling molecules on immune cells to drive immune responses towards the desired direction. Advances in our understanding of the human immune system, along with valuable lessons learned from the first generation of therapeutic biologics, are aiding the design of the next generation of immunomodulatory biologics with better therapeutic efficacy, minimized adverse effects and long-lasting clinical benefit. The recent encouraging results from antibodies targeting programmed cell death protein 1 (PD1) and B7 homolog 1 (B7H1; also known as PDL1) for the treatment of various advanced human cancers show that immunomodulatory therapy has come of age. PMID:23370250

  3. THE PROSPECTS OF THE COMBINED TREATMENT OF NON-SMALL CELL LUNGCANCER SPREAD FORMS

    Directory of Open Access Journals (Sweden)

    Akhat Bukenov

    2014-05-01

    Full Text Available The Purpose of this study is to demonstrate the effectiveness of combined treatment for III A-B stage of non-small-cell lung cancer. Non-small-cell lung cancer spread form treatment is disputable. Operation is not final solution. The combined treatment is being tested (in clinical trials in this study. This article argues that a combination of treatment is a better option than that of a specific treatment. However, there is a need to find new effective options for combined treatment.

  4. Combining inkjet printing and sol-gel chemistry for making pH-sensitive surfaces.

    Science.gov (United States)

    Orsi, Gianni; De Maria, Carmelo; Montemurro, Francesca; Chauhan, Veeren M; Aylott, Jonathan W; Vozzi, Giovanni

    2015-01-01

    Today biomedical sciences are experiencing the importance of imaging biological parameters with luminescence methods. Studying 2D pH distribution with those methods allows building knowledge about complex cellular processes. Immobilizing pH sensitive nanoparticles inside hydrogel matrixes, in order to guarantee a proper SNR, could easily make stable and biocompatible 2D sensors. Inkjet printing is also well known as tool for printing images onto porous surfaces. Recently it has been used as a free-form fabrication method for building three-dimensional parts, and now is being explored as a way of printing electrical and optical devices. Inkjet printing was used either as a rapid prototyping method for custom biosensors. Sol-gel method is naturally bound with inkjet, because the picoliter-sized ink droplets evaporate quickly, thus allowing quick sol-gel transitions on the printed surface. In this work will be shown how to merge those technologies, in order to make a nanoparticles doped printable hydrogel, which could be used for making 2D/3D smart scaffolds able to monitor cell activities. An automated image analysis system was developed in order to quickly have the pH measurements from pH nanosensors fluorescence images. PMID:25547966

  5. Combination strategy of multi-layered surface camouflage using hyperbranched polyethylene glycol and immunosuppressive drugs for the prevention of immune reactions against transplanted porcine islets.

    Science.gov (United States)

    Haque, Muhammad R; Jeong, Jee-Heon; Byun, Youngro

    2016-04-01

    This study suggests a novel method of stabilizing fragile porcine islets to prevent the dissociation after isolation and reducing immune cell invasion in a combination therapy of 'surface camouflaging' and immunosuppressive drugs (FK506, Rapamycin, MR-1, anti-CD19 mAb, and Clodrosome(®)) to effectively alleviate overall immune reactions against xenotransplanted porcine islets. The surface camouflage of pancreatic islets using biocompatible materials improved stabilization of pancreatic islet and prevented the infiltration of immune cells. Firstly, the surface of porcine islets was camouflaged by SH-6-arm-PEG-lipid and gelatin-catechol (artificial extracellular matrix) in order to stabilize the fragile isolated islets. Secondly, three different PEG layers (6-arm-PEG-SH, 6-arm-PEG-catechol, and linear PEG-SH) were chemically conjugated onto the surface of the stabilized porcine islets. Both artificial extracellular matrix (artificial ECM) and PEGylation effectively covered the surface of porcine islets without increasing the size of the whole islet. In addition, the viability and functionality of the islets were not affected by this multi-layer surface modification. The multi-layer modification significantly reduced the attachment of human serum albumin, fibronectin, and immunoglobulin G in comparison to the control collagen surface. The combination effect of multi-layer PEGylation and cocktailed immunosuppressive drugs on the survival time of the transplanted islets was assessed in a xenogeneic porcine-to-mouse model. The median survival time (MST) of 'artificial ECM + PEGylation' group was 4-fold increased compared to that of control group. In addition, the MST of 'artificial ECM + PEGylation + drug' group was 2.16-fold increased, compared to the 'control + drug' group. In conclusion, we proposed a novel porcine islet transplantation protocol using surface multi-layer modification and cocktailed immunosuppressive drugs, for stabilization and

  6. Combination of verteporfin-PDT and PI3K inhibitors enhances cell growth inhibition and apoptosis in endothelial cells

    Science.gov (United States)

    Kraus, Daniel; Chen, Bin

    2016-03-01

    Vascular targeted photodynamic therapy is a promising cancer treatment modality by ablating tumor vasculature. The effectiveness of this treatment is often compromised by regrowth of endothelial cells, which causes tumor recurrence. In this preliminary report, we showed that activated PI3K signaling was involved in endothelial cell regrowth after PDT with verteporfin and combination between verteporfin-PDT and PI3K pathway inhibitor BEZ235 induced more cell apoptosis and greater inhibition in cell proliferation. These results suggest that rational combination of verteporfin-PDT and PI3K inhibitors result in enhanced treatment outcomes.

  7. Radioresistance of granulation tissue-derived cells from skin wounds combined with total body irradiation.

    Science.gov (United States)

    Dai, Tingyu; Chen, Zelin; Tan, Li; Shi, Chunmeng

    2016-04-01

    Combined radiation and wound injury (CRWI) occurs following nuclear explosions and accidents, radiological or nuclear terrorism, and radiation therapy combined with surgery. CRWI is complicated and more difficult to heal than single injuries. Stem cell‑based therapy is a promising treatment strategy for CRWI, however, sourcing stem cells remains a challenge. In the present study, the granulation tissue-derived cells (GTCs) from the skin wounds (SWs) of CRWI mice (C‑GTCs) demonstrated a higher radioresistance to the damage caused by combined injury, and were easier to isolate and harvest when compared with bone marrow‑derived mesenchymal stromal cells (BMSCs). Furthermore, the C-GTCs exhibited similar stem cell-associated properties, such as self-renewal and multilineage differentiation capacity, when compared with neonatal dermal stromal cells (DSCs) and GTCs from unirradiated SWs. Granulation tissue, which is easy to access, may present as an optimal autologous source of stem/progenitor cells for therapeutic applications in CRWI. PMID:26936439

  8. RPE cell surface proteins in normal and dystrophic rats

    International Nuclear Information System (INIS)

    Membrane-bound proteins in plasma membrane enriched fractions from cultured rat RPE were analyzed by two-dimensional gel electrophoresis. Membrane proteins were characterized on three increasingly specific levels. Total protein was visualized by silver staining. A maximum of 102 separate proteins were counted in silver-stained gels. Glycoproteins were labeled with 3H-glucosamine or 3H-fucose and detected by autoradiography. Thirty-eight fucose-labeled and 61-71 glucosamine-labeled proteins were identified. All of the fucose-labeled proteins were labeled with glucosamine-derived radioactivity. Proteins exposed at the cell surface were labeled by lactoperoxidase-catalyzed radioiodination prior to preparation of membranes for two-dimensional analysis. Forty separate 125I-labeled surface proteins were resolved by two-dimensional electrophoresis/autoradiography. Comparison with the glycoprotein map showed that a number of these surface labeled proteins were glycoproteins. Two-dimensional maps of total protein, fucose-labeled, and glucosamine-labeled glycoproteins, and 125I-labeled surface proteins of membranes from dystrophic (RCS rdy-p+) and normal (Long Evans or RCS rdy+p+) RPE were compared. No differences in the total protein or surface-labeled proteins were observed. However, the results suggest that a 183K glycoprotein is more heavily glycosylated with glucosamine and fucose in normal RPE membranes as compared to membranes from dystrophic RPE

  9. RPE cell surface proteins in normal and dystrophic rats

    Energy Technology Data Exchange (ETDEWEB)

    Clark, V.M.; Hall, M.O.

    1986-02-01

    Membrane-bound proteins in plasma membrane enriched fractions from cultured rat RPE were analyzed by two-dimensional gel electrophoresis. Membrane proteins were characterized on three increasingly specific levels. Total protein was visualized by silver staining. A maximum of 102 separate proteins were counted in silver-stained gels. Glycoproteins were labeled with 3H-glucosamine or 3H-fucose and detected by autoradiography. Thirty-eight fucose-labeled and 61-71 glucosamine-labeled proteins were identified. All of the fucose-labeled proteins were labeled with glucosamine-derived radioactivity. Proteins exposed at the cell surface were labeled by lactoperoxidase-catalyzed radioiodination prior to preparation of membranes for two-dimensional analysis. Forty separate 125I-labeled surface proteins were resolved by two-dimensional electrophoresis/autoradiography. Comparison with the glycoprotein map showed that a number of these surface labeled proteins were glycoproteins. Two-dimensional maps of total protein, fucose-labeled, and glucosamine-labeled glycoproteins, and 125I-labeled surface proteins of membranes from dystrophic (RCS rdy-p+) and normal (Long Evans or RCS rdy+p+) RPE were compared. No differences in the total protein or surface-labeled proteins were observed. However, the results suggest that a 183K glycoprotein is more heavily glycosylated with glucosamine and fucose in normal RPE membranes as compared to membranes from dystrophic RPE.

  10. Combination therapy targeting both cancer stem-like cells and bulk tumor cells for improved efficacy of breast cancer treatment.

    Science.gov (United States)

    Wang, Tao; Narayanaswamy, Radhika; Ren, Huilan; Torchilin, Vladimir P

    2016-06-01

    Many types of tumors are organized in a hierarchy of heterogeneous cell populations. The cancer stem-like cells (CSCs) hypothesis suggests that tumor development and metastasis are driven by a minority population of cells, which are responsible for tumor initiation, growth and recurrences. The inability to efficiently eliminate CSCs during chemotherapy, together with CSCs being highly tumorigenic and invasive, may result in treatment failure due to cancer relapse and metastases. CSCs are emerging as a promising target for the development of translational cancer therapies. Ideal panacea for cancer would kill all malignant cells, including CSCs and bulk tumor cells. Since both chemotherapy and CSCs-specific therapy are insufficient to cure cancer, we propose combination therapy with CSCs-targeted agents and chemotherapeutics for improved breast cancer treatment. We generated in vitro mammosphere of 2 breast cancer cell lines, and demonstrated ability of mammospheres to grow and enrich cancer cells with stem-like properties, including self-renewal, multilineage differentiation and enrichment of cells expressing breast cancer stem-like cell biomarkers CD44(+)/CD24(-/low). The formation of mammospheres was significantly inhibited by salinomycin, validating its pharmacological role against the cancer stem-like cells. In contrast, paclitaxel showed a minimal effect on the proliferation and growth of breast cancer stem-like cells. While combination therapies of salinomycin with conventional chemotherapy (paclitaxel or lipodox) showed a potential to improve tumor cell killing, different subtypes of breast cancer cells showed different patterns in response to the combination therapies. While optimization of combination therapy is warranted, the design of combination therapy should consider phenotypic attributes of breast cancer types. PMID:27259361

  11. Combination therapy targeting both cancer stem-like cells and bulk tumor cells for improved efficacy of breast cancer treatment.

    Science.gov (United States)

    Wang, Tao; Narayanaswamy, Radhika; Ren, Huilan; Torchilin, Vladimir P

    2016-06-01

    Many types of tumors are organized in a hierarchy of heterogeneous cell populations. The cancer stem-like cells (CSCs) hypothesis suggests that tumor development and metastasis are driven by a minority population of cells, which are responsible for tumor initiation, growth and recurrences. The inability to efficiently eliminate CSCs during chemotherapy, together with CSCs being highly tumorigenic and invasive, may result in treatment failure due to cancer relapse and metastases. CSCs are emerging as a promising target for the development of translational cancer therapies. Ideal panacea for cancer would kill all malignant cells, including CSCs and bulk tumor cells. Since both chemotherapy and CSCs-specific therapy are insufficient to cure cancer, we propose combination therapy with CSCs-targeted agents and chemotherapeutics for improved breast cancer treatment. We generated in vitro mammosphere of 2 breast cancer cell lines, and demonstrated ability of mammospheres to grow and enrich cancer cells with stem-like properties, including self-renewal, multilineage differentiation and enrichment of cells expressing breast cancer stem-like cell biomarkers CD44(+)/CD24(-/low). The formation of mammospheres was significantly inhibited by salinomycin, validating its pharmacological role against the cancer stem-like cells. In contrast, paclitaxel showed a minimal effect on the proliferation and growth of breast cancer stem-like cells. While combination therapies of salinomycin with conventional chemotherapy (paclitaxel or lipodox) showed a potential to improve tumor cell killing, different subtypes of breast cancer cells showed different patterns in response to the combination therapies. While optimization of combination therapy is warranted, the design of combination therapy should consider phenotypic attributes of breast cancer types.

  12. Development of living cell force sensors for the interrogation of cell surface interactions

    Science.gov (United States)

    Brown, Scott Chang

    The measurement of cell surface interactions, or cell interaction forces, are critical for the early diagnosis and prevention of disease, the design of targeted drug and gene delivery vehicles, the development of next-generation implant materials, and much more. However, the technologies and devices that are currently available are highly limited with respect to the dynamic force range over which they can measure cell-cell or cell-substratum interactions, and with their ability to adequately mimic biologically relevant systems. Consequently, research efforts that involve cell surface interactions have been limited. In this dissertation, existing tools for research at the nanoscale (i.e., atomic force microscopy microcantilevers) are modified to develop living cell force sensors that allow for the highly sensitive measurement of cell-mediated interactions over the entire range of forces expected in biotechnology (and nano-biotechnology) research (from a single to millions of receptor-ligand bonds). Several force sensor motifs have been developed that can be used to measure interactions using single adherent cells, single suspension culture cell, and cell monolayers (tissues) over a wide range of interaction conditions (e.g., approach velocity, shear rate, contact time) using a conventional atomic force microscope. This new tool has been applied to study the pathogenesis of spontaneous pneumothorax and the interaction of cells with 14 man-made interfaces. Consequently, a new hypothesis of the interactions that manifest spontaneous pneumothorax has been developed. Additionally, these findings have the potential to lead to the development of tools for data mining materials and surfaces for unique cell interactions that could have an immense societal impact.

  13. Short-chain ceramides depress integrin cell surface expression and function in colorectal cancer cells.

    Science.gov (United States)

    Morad, Samy A F; Bridges, Lance C; Almeida Larrea, Alex D; Mayen, Anthony L; MacDougall, Matthew R; Davis, Traci S; Kester, Mark; Cabot, Myles C

    2016-07-01

    Colorectal cancer (CRC) is highly metastatic, significantly so to liver, a characteristic that embodies one of the most challenging aspects of treatment. The integrin family of cell-cell and cell-matrix adhesion receptors plays a central role in migration and invasion, functions that underlie metastatic potential. In the present work we sought to determine the impact of ceramide, which plays a key modulatory role in cancer suppression, on integrin cell surface expression and function in CRC cells in order to reveal possible ceramide-centric effects on tumor cell motility. Human CRC cells LoVo, HT-29, and HCT-116 were employed, which represent lines established from primary and metastatic sites. A cell-permeable, short-chain analog, C6-ceramide, was used as ceramide mimic. Exposure of cells to C6-ceramide (24 h) promoted a dose-dependent (2.5-10 µM) decrease in the expression of cell surface β1 and β4 integrin subunits in all cell lines; at 10 µM C6-ceramide, the decreases ranged from 30 to 50% of the control. Expression of cell surface αVβ6 integrin, which is associated with advanced invasion in CRC, was also suppressed by C6-ceramide. Decreases in integrin expression translated to diminished cellular adhesion, 50% of the control at 5 µM C6-ceramide, and markedly reduced cellular migration, approximately 30-40% of the control in all cell lines. Physicochemical examination revealed potent efficacy of nano-formulated C6-ceramide, but inferior activity of dihydro-C6-ceramide and L-C6-ceramide, compared to the unsaturated counterpart and the natural d-enantiomer, respectively. These studies demonstrate novel actions of ceramides that may have application in suppression of tumor metastasis, in addition to their known tumor suppressor effects. PMID:27045476

  14. Analysis of the Interactions of Botanical Extract Combinations Against the Viability of Prostate Cancer Cell Lines

    Directory of Open Access Journals (Sweden)

    Lynn S. Adams

    2006-01-01

    Full Text Available Herbal medicines are often combinations of botanical extracts that are assumed to have additive or synergistic effects. The purpose of this investigation was to compare the effect of individual botanical extracts with combinations of extracts on prostate cell viability. We then modeled the interactions between botanical extracts in combination isobolographically. Scutellaria baicalensis, Rabdosia rubescens, Panax-pseudo ginseng, Dendranthema morifolium, Glycyrrhiza uralensis and Serenoa repens were collected, taxonomically identified and extracts prepared. Effects of the extracts on cell viability were quantitated in prostate cell lines using a luminescent ATP cell viability assay. Combinations of two botanical extracts of the four most active extracts were tested in the 22Rv1 cell line and their interactions assessed using isobolographic analysis. Each extract significantly inhibited the proliferation of prostate cell lines in a time- and dose-dependent manner except repens. The most active extracts, baicalensis, D. morifolium, G. uralensis and R. rubescens were tested as two-extract combinations. baicalensis and D. morifolium when combined were additive with a trend toward synergy, whereas D. morifolium and R. rubescens together were additive. The remaining two-extract combinations showed antagonism. The four extracts together were significantly more effective than the two-by-two combinations and the individual extracts alone. Combining the four herbal extracts significantly enhanced their activity in the cell lines tested compared with extracts alone. The less predictable nature of the two-way combinations suggests a need for careful characterization of the effects of each individual herb based on their intended use.

  15. Functionalized PDMS with versatile and scalable surface roughness gradients for cell culture

    KAUST Repository

    Zhou, Bingpu

    2015-07-21

    This manuscript describes a simple and versatile approach to engineering surface roughness gradients via combination of microfluidics and photo-polymerization. Through UV-mediated polymerization, N-isopropylacrylamide with concentration gradients are successfully grafted onto PDMS surface, leading to diverse roughness degrees on the obtained PDMS substrate. Furthermore, the extent of surface roughness can be controllably regulated via tuning the flow rate ratio between the monomer solution and deionized water. Average roughness ranging from 8.050 nm to 151.68 nm has well been achieved in this work. Such PDMS samples are also demonstrated to be capable of working as supporting substrates for controlling cell adhesion or detachment. Due to the different degrees of surface roughness on a single substrate, our method provides an effective approach for designing advanced surafecs for cell culture. Finally, the thermosensitive property of N-isopropylacrylamide makes our sample furnish as another means for controlling the cell detachment from the substrates with correspondence to the surrounding temperature.

  16. Fibronectin on the Surface of Myeloma Cell-derived Exosomes Mediates Exosome-Cell Interactions.

    Science.gov (United States)

    Purushothaman, Anurag; Bandari, Shyam Kumar; Liu, Jian; Mobley, James A; Brown, Elizabeth E; Sanderson, Ralph D

    2016-01-22

    Exosomes regulate cell behavior by binding to and delivering their cargo to target cells; however, the mechanisms mediating exosome-cell interactions are poorly understood. Heparan sulfates on target cell surfaces can act as receptors for exosome uptake, but the ligand for heparan sulfate on exosomes has not been identified. Using exosomes isolated from myeloma cell lines and from myeloma patients, we identify exosomal fibronectin as a key heparan sulfate-binding ligand and mediator of exosome-cell interactions. We discovered that heparan sulfate plays a dual role in exosome-cell interaction; heparan sulfate on exosomes captures fibronectin, and on target cells it acts as a receptor for fibronectin. Removal of heparan sulfate from the exosome surface releases fibronectin and dramatically inhibits exosome-target cell interaction. Antibody specific for the Hep-II heparin-binding domain of fibronectin blocks exosome interaction with tumor cells or with marrow stromal cells. Regarding exosome function, fibronectin-mediated binding of exosomes to myeloma cells activated p38 and pERK signaling and expression of downstream target genes DKK1 and MMP-9, two molecules that promote myeloma progression. Antibody against fibronectin inhibited the ability of myeloma-derived exosomes to stimulate endothelial cell invasion. Heparin or heparin mimetics including Roneparstat, a modified heparin in phase I trials in myeloma patients, significantly inhibited exosome-cell interactions. These studies provide the first evidence that fibronectin binding to heparan sulfate mediates exosome-cell interactions, revealing a fundamental mechanism important for exosome-mediated cross-talk within tumor microenvironments. Moreover, these results imply that therapeutic disruption of fibronectin-heparan sulfate interactions will negatively impact myeloma tumor growth and progression.

  17. Interaction of KSHV with Host Cell Surface Receptors and Cell Entry

    Directory of Open Access Journals (Sweden)

    Mohanan Valiya Veettil

    2014-10-01

    Full Text Available Virus entry is a complex process characterized by a sequence of events. Since the discovery of KSHV in 1994, tremendous progress has been made in our understanding of KSHV entry into its in vitro target cells. KSHV entry is a complex multistep process involving viral envelope glycoproteins and several cell surface molecules that is utilized by KSHV for its attachment and entry. KSHV has a broad cell tropism and the attachment and receptor engagement on target cells have an important role in determining the cell type-specific mode of entry. KSHV utilizes heparan sulfate, integrins and EphrinA2 molecules as receptors which results in the activation of host cell pre-existing signal pathways that facilitate the subsequent cascade of events resulting in the rapid entry of virus particles, trafficking towards the nucleus followed by viral and host gene expression. KSHV enters human fibroblast cells by dynamin dependant clathrin mediated endocytosis and by dynamin independent macropinocytosis in dermal endothelial cells. Once internalized into endosomes, fusion of the viral envelope with the endosomal membranes in an acidification dependent manner results in the release of capsids which subsequently reaches the nuclear pore vicinity leading to the delivery of viral DNA into the nucleus. In this review, we discuss the principal mechanisms that enable KSHV to interact with the host cell surface receptors as well as the mechanisms that are required to modulate cell signaling machinery for a successful entry.

  18. Distribution of anionic groups at the cell surface of different Sporothrix schenckii cell types.

    Science.gov (United States)

    Benchimol, M; de Souza, W; Travassos, L R

    1979-06-01

    The distribution of anionic groups at the cell surface of yeastlike forms, hyphae, and conidia of Sporothrix schenckii was studied by staining with colloidal iron hydroxide and cationized ferritin. By using colloidal iron hydroxide it was shown that the external cell wall layer of one strain (strain 1099.18) could be resolved into two reactive sublayers and that these layers were present in many but not all cells of the same population. In contrast, most cells of another strain (strain 1099.12) were stained by colloidal iron hydroxide, but only one reactive layer was seen. Acidic layers of the yeastlike forms of the two strains were much thicker than those of conidia and hyphae. By the cationized ferritin staining procedure it was observed that the acidic layers of yeast forms sloughed off of cells, probably due to cell-cell or cell-medium attrition in shaken submerged cultures or to a process by which the outer layers detach from cells as they are replaced by newly synthesized ones. The colloidal iron hydroxide- and cationized ferritin-reactive cell surface layers of S. schenckii correspond to the previously described (L. R. Travassos et al., Exp. Mycol. 1:293-305, 1977) concanavalin A-reactive peptidorhamnomannan complexes, and their reactivity is probably due to the presence of acidic amino acids of low pK values rather than to glucuronic acid units.

  19. Combination of ionising irradiation and hyperthermia activates programmed apoptotic and necrotic cell death pathways in human colorectal carcinoma cells

    Energy Technology Data Exchange (ETDEWEB)

    Mantel, Frederick; Frey, Benjamin; Haslinger, Stefan [Erlangen-Nuernberg Univ., University Hospital Erlangen (DE). Dept. of Radiation Oncology] (and others)

    2010-11-15

    The malignancy of tumor cells can be attenuated by interfering with cell death pathways. Since hyperthermia (HT) is a very potent radiosensitizer, the influence of HT (41.5 C for 1 hour) alone and in combination with ionising irradiation (X-ray; 5 Gy or 10 Gy) on the form of cell death as well as on the expression of proteins known to be major components in tumor cells' apoptotic and necrotic pathways were examined in colorectal tumor cells. The expression of proteins was analysed by western blot and the relative activity of caspases-3/7 by fluorescence- based assay. Colony formation was analysed using the clonogenic assay and cell death was determined with annexin V-FITC/propidium iodide staining. Combining X-ray with HT led to similar activation of caspase-3/7 and p53 expression in comparison to irradiation only while the amount of the pro-apoptotic proteins PUMA and Bax was increased in HCT15 and SW480 cells. HT alone or combinations with X-ray further resulted in a temporarily increased level of the anti-apoptotic protein Bcl-2. Irradiation plus HT further led to an up-regulation of IRF-5. The levels of RIP-1, a marker for programmed necrosis, increased in tumor cells which were treated with HT and/or X-ray. Combining 5 Gy irradiation with HT compared to irradiation resulted in a significantly increased number of necrotic tumor cells and in decreased colony formation. The combined treatment of colorectal tumor cells with X-ray and HT activates distinct tumor cell pathways and fosters the early appearance of a necrotic tumor cell phenotype. (orig.)

  20. Investigation of the effects of rear surface recombination on the Cu(In,Ga)Se2 solar cell performances

    Science.gov (United States)

    Umehara, Takeshi; Iinuma, Shohei; Yamada, Akira

    2016-07-01

    This study investigated the band profile design of Cu(In,Ga)Se2 (CIGS) solar cells by considering the rear surface recombination. We compared the structures assuming the back surface field (BSF), passivation and graded band profile by using device simulator. As a result, it was found that the band structure of a combination of a flat-band and a single-graded profile is the suitable structure for CIGS solar cells with the absorber thickness of around 1.0 μm. In addition, the back passivation technique is unnecessary in the case of CIGS solar cells with a band profiling technique. We proposed that the band structure of a combination of a flat-band and a single-graded profile is the most practical and effective way for CIGS solar cells. [Figure not available: see fulltext.

  1. Arctigenin in combination with quercetin synergistically enhances the anti-proliferative effect in prostate cancer cells

    Science.gov (United States)

    Wang, Piwen; Phan, Tien; Gordon, David; Chung, Seyung; Henning, Susanne M.; Vadgama, Jaydutt V.

    2014-01-01

    Scope We investigated whether a combination of two promising chemopreventive agents arctigenin and quercetin increases the anti-carcinogenic potency at lower concentrations than necessary when used individually in prostate cancer. Methods and results Androgen-dependent LAPC-4 and LNCaP prostate cancer cells were treated with low doses of arctigenin and quercetin alone or in combination for 48h. The anti-proliferative activity of arctigenin was 10-20 fold stronger than quercetin in both cell lines. Their combination synergistically enhanced the anti-proliferative effect, with a stronger effect in androgen receptor (AR) wild-type LAPC-4 cells than in AR mutated LNCaP cells. Arctigenin demonstrated a strong ability to inhibit AR protein expression in LAPC-4 cells. The combination treatment significantly inhibited both AR and PI3K/Akt pathways compared to control. A protein array analysis revealed that the mixture targets multiple pathways particularly in LAPC-4 cells including Stat3 pathway. The mixture significantly inhibited the expression of several oncogenic microRNAs including miR-21, miR-19b, and miR-148a compared to control. The mixture also enhanced the inhibition of cell migration in both cell lines compared to individual compounds tested. Conclusion The combination of arctigenin and quercetin, that target similar pathways, at low physiological doses, provides a novel regimen with enhanced chemoprevention in prostate cancer. PMID:25380086

  2. Chemical characterization and dynamics of particulate combined amino acids in Pacific surface waters

    Science.gov (United States)

    Tsukasaki, Ayumi; Tanoue, Eiichiro

    2010-01-01

    Particulate organic matter (POM) in surface water plays an essential role in maintaining marine ecosystem and is a complex mixture of living POM (organisms) and non-living POM (detritus). Transformation of organic constituents of organisms to detritus may be the most active part in the decomposition processes of organic matter in marine biogeochemistry. Distribution and chemical fractionation of particulate combined amino acids (PCAAs) in surface waters from Southern Ocean to tropical waters along a transect of 170°W were examined to clarify the transfer processes of organic constituents of organisms to detritus. The observational facts may be summarized as follows. 1) Degradation Index (DI) based on principal component analysis of amino acid composition of bulk PCAAs agreed with the traditional degradation indicators of bulk POM, i.e., PCAAs-carbon (C)/particulate organic carbon (POC), PCAAs-nitrogen (N)/particulate nitrogen (PN) and C/N. 2) PCAAs were distinguished into 3 fractions, i.e., Urea-soluble, Sodium Dodecyl Sulfate (SDS)-soluble and SDS/Urea-insoluble fractions, by means of solubility against reagents frequently used for protein extraction from biological samples. Proportions of PCAAs of the 3 fractions to bulk PCAAs were almost constant, although concentrations of the PCAAs of 3 fractions as well as bulk in seawater varied two orders of magnitude over the transect. 3) There was no significant difference among DI values of the 3 fractions, although molecular mass distributions of the Urea-soluble and SDS-soluble fractions determined by SDS-polyacrylamide gel electrophoresis were different. The results indicated that no one fraction was particularly susceptible to degradation and that there is a common mechanism for transferring and maintaining the similar chemical quality of bulk POM over the transect.

  3. Surface Partial Discharge Characteristics of Oil-paper Insulation Under Combined AC-DC Voltage

    Institute of Scientific and Technical Information of China (English)

    SHA Yanchao; ZHOU Yuanxiang; NIE Dexin; WU Zhirong; DENG Jiangang; LU Licheng

    2013-01-01

    The valve side windings of converter transformers bear AC,DC,impulse,and reversal-polarity voltages during operation,which could result in serious insulation problems of the equipment.By performing experiments with surface discharge model of oil-paper insulation at 80 ℃ under combined AC-DC voltage for 200 h,we studied the spectrums and statistical parameters of partial discharges at different discharge stages.Furthermore,some fingerprint parameters were calculated in order to estimate the development situation of partial discharge,while the characteristic gases dissolved in the transformer oil were measured by gas chromatography.The surface discharges in the experiments were observed using a high speed camera,and a full discharge process could be marked off into four stages as follows.①The elementary stage.When a partial discharge occurs near electrodes,electrical charges are injected into the region near electrodes and causing bubble generation.②Due to their high resistivity and low dielectric constant,the bubbles would bare the major part of the voltage applied to samples.Therefore,discharge happens inside the small bubbles,and it emits a lot of light.③Micmmolecules of gas are produced in discharge,and further ionization in the transformer oil takes place simultaneously when high-energy electrons collide with oil molecules.④The carrier charge moves forward to electrodes driven by the applied electric field,till they neutralize with the charge from electrodes,and hence discharge channels are formed subsequently.

  4. Detecting Bacterial Surface Organelles on Single Cells Using Optical Tweezers.

    Science.gov (United States)

    Zakrisson, Johan; Singh, Bhupender; Svenmarker, Pontus; Wiklund, Krister; Zhang, Hanqing; Hakobyan, Shoghik; Ramstedt, Madeleine; Andersson, Magnus

    2016-05-10

    Bacterial cells display a diverse array of surface organelles that are important for a range of processes such as intercellular communication, motility and adhesion leading to biofilm formation, infections, and bacterial spread. More specifically, attachment to host cells by Gram-negative bacteria are mediated by adhesion pili, which are nanometers wide and micrometers long fibrous organelles. Since these pili are significantly thinner than the wavelength of visible light, they cannot be detected using standard light microscopy techniques. At present, there is no fast and simple method available to investigate if a single cell expresses pili while keeping the cell alive for further studies. In this study, we present a method to determine the presence of pili on a single bacterium. The protocol involves imaging the bacterium to measure its size, followed by predicting the fluid drag based on its size using an analytical model, and thereafter oscillating the sample while a single bacterium is trapped by an optical tweezer to measure its effective fluid drag. Comparison between the predicted and the measured fluid drag thereby indicate the presence of pili. Herein, we verify the method using polymer coated silica microspheres and Escherichia coli bacteria expressing adhesion pili. Our protocol can in real time and within seconds assist single cell studies by distinguishing between piliated and nonpiliated bacteria. PMID:27088225

  5. A reflectometer for the combined measurement of refractive index, microroughness, macroroughness and gloss of low-extinction surfaces

    Science.gov (United States)

    Elton, N. J.; Day, J. C. C.

    2009-02-01

    Roughness, porosity and gloss are important properties of many surfaces including coated and printed paper, paints and mineral-filled plastics. A visible light reflectometer is described for essentially simultaneous measurement of refractive index, gloss, microroughness (rms amplitude in the sub-wavelength region) and the two-dimensional forward scattering pattern (yielding the surface slope distribution and associated statistics) of glossy or semi-glossy low-extinction surfaces. The principal novelty is the ability to obtain this useful combination of data in a single measurement. In combination with an x-y stage, the reflectometer can readily produce surface maps of the various measurement parameters and it is expected that the data should offer improved insights into cause and effect in paper and other surfaces, for example to identify causes of gloss and print mottle. Illustrative experimental data are provided and some comparisons made between data obtained by reflectometry and by alternative techniques such as spectroscopic ellipsometry and atomic force microscopy.

  6. Hyperbaric oxygen therapy combined with Schwann cell transplantation promotes spinal cord injury recovery

    Directory of Open Access Journals (Sweden)

    Chuan-gang Peng

    2015-01-01

    Full Text Available Schwann cell transplantation and hyperbaric oxygen therapy each promote recovery from spinal cord injury, but it remains unclear whether their combination improves therapeutic results more than monotherapy. To investigate this, we used Schwann cell transplantation via the tail vein, hyperbaric oxygen therapy, or their combination, in rat models of spinal cord contusion injury. The combined treatment was more effective in improving hindlimb motor function than either treatment alone; injured spinal tissue showed a greater number of neurite-like structures in the injured spinal tissue, somatosensory and motor evoked potential latencies were notably shorter, and their amplitudes greater, after combination therapy than after monotherapy. These findings indicate that Schwann cell transplantation combined with hyperbaric oxygen therapy is more effective than either treatment alone in promoting the recovery of spinal cord in rats after injury.

  7. Hyperbaric oxygen therapy combined with Schwann cell transplantation promotes spinal cord injury recover y

    Institute of Scientific and Technical Information of China (English)

    Chuan-gang Peng; Shu-quan Zhang; Min-fei Wu; Yang Lv; Dan-kai Wu; Qi Yang; Rui Gu

    2015-01-01

    Schwann cell transplantation and hyperbaric oxygen therapy each promote recovery from spinal cord injury, but it remains unclear whether their combination improves therapeutic results more than monotherapy. To investigate this, we used Schwann cell transplantationviathe tail vein, hyperbaric oxygen therapy, or their combination, in rat models of spinal cord contusion injury. The combined treatment was more effective in improving hindlimb motor function than either treatment alone; injured spinal tissue showed a greater number of neurite-like structures in the injured spinal tissue, somatosensory and motor evoked potential latencies were notably shorter, and their amplitudes greater, after combination therapy than after monotherapy. These ifndings indicate that Schwann cell transplantation combined with hyperbaric oxygen therapy is more effective than either treatment alone in promoting the recovery of spinal cord in rats after injury.

  8. Cell surface profiling using high-throughput flow cytometry: a platform for biomarker discovery and analysis of cellular heterogeneity.

    Directory of Open Access Journals (Sweden)

    Craig A Gedye

    Full Text Available Cell surface proteins have a wide range of biological functions, and are often used as lineage-specific markers. Antibodies that recognize cell surface antigens are widely used as research tools, diagnostic markers, and even therapeutic agents. The ability to obtain broad cell surface protein profiles would thus be of great value in a wide range of fields. There are however currently few available methods for high-throughput analysis of large numbers of cell surface proteins. We describe here a high-throughput flow cytometry (HT-FC platform for rapid analysis of 363 cell surface antigens. Here we demonstrate that HT-FC provides reproducible results, and use the platform to identify cell surface antigens that are influenced by common cell preparation methods. We show that multiple populations within complex samples such as primary tumors can be simultaneously analyzed by co-staining of cells with lineage-specific antibodies, allowing unprecedented depth of analysis of heterogeneous cell populations. Furthermore, standard informatics methods can be used to visualize, cluster and downsample HT-FC data to reveal novel signatures and biomarkers. We show that the cell surface profile provides sufficient molecular information to classify samples from different cancers and tissue types into biologically relevant clusters using unsupervised hierarchical clustering. Finally, we describe the identification of a candidate lineage marker and its subsequent validation. In summary, HT-FC combines the advantages of a high-throughput screen with a detection method that is sensitive, quantitative, highly reproducible, and allows in-depth analysis of heterogeneous samples. The use of commercially available antibodies means that high quality reagents are immediately available for follow-up studies. HT-FC has a wide range of applications, including biomarker discovery, molecular classification of cancers, or identification of novel lineage specific or stem cell

  9. Hybrid silicon honeycomb/organic solar cells with enhanced efficiency using surface etching

    Science.gov (United States)

    Liu, Ruiyuan; Sun, Teng; Liu, Jiawei; Wu, Shan; Sun, Baoquan

    2016-06-01

    Silicon (Si) nanostructure-based photovoltaic devices are attractive for their excellent optical and electrical performance, but show lower efficiency than their planar counterparts due to the increased surface recombination associated with the high surface area and roughness. Here, we demonstrate an efficiency enhancement for hybrid nanostructured Si/polymer solar cells based on a novel Si honeycomb (SiHC) structure using a simple etching method. SiHC structures are fabricated using a combination of nanosphere lithography and plasma treatment followed by a wet chemical post-etching. SiHC has shown superior light-trapping ability in comparison with the other Si nanostructures, along with a robust structure. Anisotropic tetramethylammonium hydroxide etching not only tunes the final surface morphologies of the nanostructures, but also reduces the surface roughness leading to a lower recombination rate in the hybrid solar cells. The suppressed recombination loss, benefiting from the reduced surface-to-volume ratio and roughness, has resulted in a high open-circuit voltage of 600 mV, a short-circuit current of 31.46 mA cm-2 due to the light-trapping ability of the SiHCs, and yields a power conversion efficiency of 12.79% without any other device structure optimization.

  10. Yeast cell surface display for lipase whole cell catalyst and its applications

    Energy Technology Data Exchange (ETDEWEB)

    Liu, Yun; Zhang, Rui; Lian, Zhongshuai; Wang, Shihui; Wright, Aaron T.

    2014-08-01

    The cell surface display technique allows for the expression of target proteins or peptides on the microbial cell surface by fusing an appropriate protein as an anchoring motif. Yeast display systems, such as Pichia pastoris, Yarowia lipolytica and Saccharomyces cerevisiae, are ideal, alternative and extensive display systems with the advantage of simple genetic manipulation and post-translational modification of expressed heterologous proteins. Engineered yeasts show high performance characteristics and variant utilizations. Herein, we comprehensively summarize the variant factors affecting lipase whole cell catalyst activity and display efficiency, including the structure and size of target proteins, screening anchor proteins, type and chain length of linkers, and the appropriate matching rules among the above-mentioned display units. Furthermore, we also address novel approaches to enhance stability and activity of recombinant lipases, such as VHb gene co-expression, multi-enzyme co-display technique, and the micro-environmental interference and self-assembly techniques. Finally, we represent the variety of applications of whole cell surface displayed lipases on yeast cells in non-aqueous phases, including synthesis of esters, PUFA enrichment, resolution of chiral drugs, organic synthesis and biofuels. We demonstrate that the lipase surface display technique is a powerful tool for functionalizing yeasts to serve as whole cell catalysts, and increasing interest is providing an impetus for broad application of this technique.

  11. "Race for the Surface": Eukaryotic Cells Can Win.

    Science.gov (United States)

    Pham, Vy T H; Truong, Vi Khanh; Orlowska, Anna; Ghanaati, Shahram; Barbeck, Mike; Booms, Patrick; Fulcher, Alex J; Bhadra, Chris M; Buividas, Ričardas; Baulin, Vladimir; Kirkpatrick, C James; Doran, Pauline; Mainwaring, David E; Juodkazis, Saulius; Crawford, Russell J; Ivanova, Elena P

    2016-08-31

    With an aging population and the consequent increasing use of medical implants, managing the possible infections arising from implant surgery remains a global challenge. Here, we demonstrate for the first time that a precise nanotopology provides an effective intervention in bacterial cocolonization enabling the proliferation of eukaryotic cells on a substratum surface, preinfected by both live Gram-negative, Pseudomonas aeruginosa, and Gram-positive, Staphylococcus aureus, pathogenic bacteria. The topology of the model black silicon (bSi) substratum not only favors the proliferation of eukaryotic cells but is biocompatible, not triggering an inflammatory response in the host. The attachment behavior and development of filopodia when COS-7 fibroblast cells are placed in contact with the bSi surface are demonstrated in the dynamic study, which is based on the use of real-time sequential confocal imaging. Bactericidal nanotopology may enhance the prospect for further development of inherently responsive antibacterial nanomaterials for bionic applications such as prosthetics and implants. PMID:27494044

  12. CpG ODN and ISCOMATRIX Adjuvant: A Synergistic Adjuvant Combination Inducing Strong T-Cell IFN-γ Responses

    Directory of Open Access Journals (Sweden)

    Michael J. McCluskie

    2013-01-01

    Full Text Available For the induction of robust humoral and cellular immune responses, a strong rationale exists to use vaccine-adjuvant combinations possessing both immune modulatory and enhanced delivery capabilities. Herein, we evaluated the combination of 2 different adjuvants, a TLR9 agonist, composed of synthetic oligodeoxynucleotides (ODN containing immunostimulatory CpG motifs (CpG, and ISCOMATRIX adjuvant (ISCOMATRIX, composed of saponin, phospholipid, and cholesterol, which possesses both immunostimulatory and delivery properties. While both individual adjuvants have been shown effective in numerous preclinical and clinical studies, it is likely that for optimal adjuvant activity a combined adjuvant approach will be necessary. Herein, using three different antigens, namely, hepatitis B surface antigen (HBsAg, ovalbumin (OVA, and influenza A haemagglutinin antigen (HA, we show in mice that some adjuvant effects of CpG and ISCOMATRIX are further enhanced if they are used in combination. In particular, with all three antigens, IFN-γ levels were greatly increased with the CpG/ISCOMATRIX combination. The ability of the CpG/ISCOMATRIX combination to induce antitumor responses when administered with OVA following administration to mice of a highly metastatic OVA-secreting tumor cell line (B16-OVA melanoma was also demonstrated. Thus the CpG/ISCOMATRIX combination may prove to be a valuable tool in the development of novel or improved vaccines.

  13. Combined biomass valorization and hydrogen production in a photoelectrochemical cell

    Science.gov (United States)

    Cha, Hyun Gil; Choi, Kyoung-Shin

    2015-04-01

    In a typical hydrogen-producing photoelectrochemical cell (PEC), water reduction at the cathode (producing hydrogen) is accompanied by water oxidation at the anode (producing oxygen). This anode reaction is, however, not kinetically favourable. Here we investigate the possibility of utilizing solar energy for biomass conversion by performing the oxidation of 5-hydroxymethylfurfural (HMF) into 2,5-furandicarboxylic acid (FDCA) at the anode of a PEC. HMF is a key intermediate in biomass conversion, and FDCA is an important monomer for the production of numerous polymers. Using 2,2,6,6-tetramethylpiperidine-1-oxyl as a mediator, we obtained a near-quantitative yield and 100% Faradaic efficiency at ambient conditions without the use of precious-metal catalysts. This reaction is also thermodynamically and kinetically more favourable than water oxidation. Our results suggest that solar-driven biomass conversion can be a viable anode reaction that has the potential to increase both the efficiency and the utility of PECs constructed for solar-fuel production.

  14. Improvement of the wear behaviour of highly-loaded components and tools by multi-combined surface treatment

    Directory of Open Access Journals (Sweden)

    Gundis Grumbt

    2014-01-01

    Full Text Available This paper introduces a novel multi-combined surface treatment consisting of the individual treatments of plasma nitriding (PN, physical vapour deposition (PVD and electron beam hardening (EBH. Using graded surface layers produced by such a combined surface treatment, it is possible to withstand the complex load conditions incurred by components and tools. It is shown, that the treatment sequences PN+EBH+PVD and EBH+PN+PVD are suitable for improving material properties. These multi-combined surface treatments lead to a significant improvement in load-supporting capacity. Critical load values of cohesive failure measured by scratch tests are tripled when compared to the individual treatment of PVD, and increased by at least 20% in comparison to the duplex treatments of EBH+PVD or PN+PVD. The metallurgical compatibility of the single treatments is essential for the success of combined treatments. Material-specific limitations are defined, which exclude failure due to crack initiation, the occurrence of retained austenite, and tempering effects. Based on the model wear-test assembly block-on-cylinder, it was proved, that the specific wear rate of multi-combined treated specimens is reduced about 20-50% while wear of counterpart components is decreased as well. The triplex surface heat treatment introduced opens up new prospects for highly-loaded components and tools.

  15. Curcumin induced nanoscale CD44 molecular redistribution and antigen-antibody interaction on HepG2 cell surface

    Energy Technology Data Exchange (ETDEWEB)

    Wang Mu [Department of Chemistry, Jinan University, 601 Huangpu Road West, Tianhe District, Guangzhou 510632 (China); Ruan Yuxia [Department of Ophthalmology, The First Affiliated Hospital, Jinan University, 601 Huangpu Road West, Tianhe District, Guangzhou 510632 (China); Xing Xiaobo; Chen Qian; Peng, Yuan [Department of Chemistry, Jinan University, 601 Huangpu Road West, Tianhe District, Guangzhou 510632 (China); Cai Jiye, E-mail: tjycai@jnu.edu.cn [Department of Chemistry, Jinan University, 601 Huangpu Road West, Tianhe District, Guangzhou 510632 (China)

    2011-07-04

    Graphical abstract: Highlights: > In this study, we investigate the changes of CD44 expression and distribution on HepG2 cells after curcumin treatment. > We find curcumin is able to change the morphology and ultrastructure of HepG2 cells. > Curcumin can reduce the expression of CD44 molecules and induce the nanoscale molecular redistribution on cell surface. > The binding force between CD44-modified AFM tip and the HepG2 cell surface decreases after curcumin-treatment. - Abstract: The cell surface glycoprotein CD44 was implicated in the progression, metastasis and apoptosis of certain human tumors. In this study, we used atomic force microscope (AFM) to monitor the effect of curcumin on human hepatocellular carcinoma (HepG2) cell surface nanoscale structure. High-resolution imaging revealed that cell morphology and ultrastructure changed a lot after being treated with curcumin. The membrane average roughness increased (10.88 {+-} 4.62 nm to 129.70 {+-} 43.72 nm) and the expression of CD44 decreased (99.79 {+-} 0.16% to 75.14 {+-} 8.37%). Laser scanning confocal microscope (LSCM) imaging showed that CD44 molecules were located on the cell membrane. The florescence intensity in control group was weaker than that in curcumin treated cells. Most of the binding forces between CD44 antibodies and untreated HepG2 cell membrane were around 120-220 pN. After being incubated with curcumin, the major forces focused on 70-150 pN (10 {mu}M curcumin-treated) and 50-120 pN (20 {mu}M curcumin-treated). These results suggested that, as result of nanoscale molecular redistribution, changes of the cell surface were in response to external treatment of curcumin. The combination of AFM and LSCM could be a powerful method to detect the distribution of cell surface molecules and interactions between molecules and their ligands.

  16. Cell surface glycopeptides from human intestinal epithelial cell lines derived from normal colon and colon adenocarcinomas

    International Nuclear Information System (INIS)

    The cell surface glycopeptides from an epithelial cell line (CCL 239) derived from normal human colon were compared with those from three cell lines (HCT-8R, HCT-15, and CaCo-2) derived independently from human colonic adenocarcinomas. Cells were incubated with D-[2-3H]mannose or L-[5,6-3H]fucose for 24 h and treated with trypsin to release cell surface components which were then digested exhaustively with Pronase and fractionated on Bio-Gel P-6 before and after treatment with endo-beta-N-acetylglucosaminidase H. The most noticeable difference between the labeled glycopeptides from the tumor and CCL 239 cells was the presence in the former of an endo-beta-N-acetylglucosaminidase H-resistant high molecular weight glycopeptide fraction which was eluted in the void volume of Bio-Gel P-6. This fraction was obtained with both labeled mannose and fucose as precursors. However, acid hydrolysis of this fraction obtained after incubation with [2-3H]mannose revealed that as much as 60-90% of the radioactivity was recovered as fucose. Analysis of the total glycopeptides (cell surface and cell pellet) obtained after incubation with [2-3H]mannose showed that from 40-45% of the radioactivity in the tumor cells and less than 10% of the radioactivity in the CCL 239 cells was recovered as fucose. After incubation of the HCT-8R cells with D-[1,6-3H]glucosamine and L-[1-14C]fucose, strong acid hydrolysis of the labeled glycopeptide fraction excluded from Bio-Gel P-6 produced 3H-labeled N-acetylglucosamine and N-acetylgalactosamine

  17. Stability and reliability of pit slopes in surface mining combined with underground mining in Tonglushan mine

    Institute of Scientific and Technical Information of China (English)

    罗一忠; 吴爱祥; 刘湘平; 王洪江

    2004-01-01

    Slope stability is of critical importance in the process of surface-underground mining combination. The influence of underground mining on pit slope stability was mainly discussed, and the self-stabilization of underground stopes was also studied. The random finite element method was used to analyze the probability of the rock mass stability degree of both pit slopes and underground stopes. Meanwhile, 3D elasto-plastic finite element method was used to research into the stress, strain and rock mass failure resulting from mining. The results of numerical simulation indicate that the mining of the underground test stope has certain influence on the stability of the pit slope, but the influence is not great. The safety factor of pit slope is decreased by 0.06, and the failure probability of the pit slope is increased by 1.84%. In addition, the strata yielding zone exists around the underground test stope. The results basically conform to the information coming from the field monitoring.

  18. Urban Climate Station Site Selection Through Combined Digital Surface Model and Sun Angle Calculations

    Science.gov (United States)

    Kidd, Chris; Chapman, Lee

    2012-01-01

    Meteorological measurements within urban areas are becoming increasingly important due to the accentuating effects of climate change upon the Urban Heat Island (UHI). However, ensuring that such measurements are representative of the local area is often difficult due to the diversity of the urban environment. The evaluation of sites is important for both new sites and for the relocation of established sites to ensure that long term changes in the meteorological and climatological conditions continue to be faithfully recorded. Site selection is traditionally carried out in the field using both local knowledge and visual inspection. This paper exploits and assesses the use of lidar-derived digital surface models (DSMs) to quantitatively aid the site selection process. This is acheived by combining the DSM with a solar model, first to generate spatial maps of sky view factors and sun-hour potential and second, to generate site-specific views of the horizon. The results show that such a technique is a useful first-step approach to identify key sites that may be further evaluated for the location of meteorological stations within urban areas.

  19. Phospholipid polymer-based antibody immobilization for cell rolling surfaces in stem cell purification system.

    Science.gov (United States)

    Mahara, Atsushi; Chen, Hao; Ishihara, Kazuhiko; Yamaoka, Tetsuji

    2014-01-01

    We previously developed an antibody-conjugated cell rolling column that successfully separates stem cell subpopulations depending on the cell surface marker density, but a large amount of the injected cells were retained in the column because of non-specific interactions. In this study, an amphiphilic copolymer, poly[2-methacryloyloxyethyl phosphorylcholine (MPC)-co-n-butyl methacrylate (nBMA)-co-N-vinyl formamide (NVf)], with phospholipid polar side groups was designed as a novel antibody-immobilizing modifier. The formamide groups in NVf units were converted to active maleimide groups. A plastic flow microfluidic chamber was coated with the copolymers, and a reduced anti-CD90 antibody was immobilized. The adipose tissue-derived stem cells isolated from the rat were injected into the flow chamber, and their rolling behavior was observed under a microscope with a high-speed camera. Non-specific cell adhesion was reduced strongly by means of this immobilization method because of the MPC unit, resulting in a high percentage of rolling cells. These results demonstrate that a surface coated with phospholipid polar groups can be used in an effective stem cell separation system based on the cell rolling process.

  20. Evaluation of Relative Yeast Cell Surface Hydrophobicity Measured by Flow Cytometry

    Directory of Open Access Journals (Sweden)

    Lisa Colling

    2005-01-01

    Full Text Available Objective: To develop an efficient method for evaluating cell surface hydrophobicity and to apply the method to demonstrate the effects of fungal growth conditions on cell surface properties.

  1. Synergistic enhancement of NK cell-mediated cytotoxicity by combination of histone deacetylase inhibitor and ionizing radiation

    International Nuclear Information System (INIS)

    The overexpression of histone deacetylase (HDAC) and a subsequent decrease in the acetylation levels of nuclear histones are frequently observed in cancer cells. Generally it was accepted that the deacetylation of histones suppressed expression of the attached genes. Therefore, it has been suggested that HDAC might contribute to the survival of cancer cells by altering the NKG2D ligands transcripts. By the way, the translational regulation of NKG2D ligands remaines unclear in cancer cells. It appears the modulation of this unclear mechanism could enhance NKG2D ligand expressions and the susceptibility of cancer cells to NK cells. Previously, it was reported that irradiation can increase the surface expressions of NKG2D ligands on several cancer cell types without increasing the levels of NKG2D ligand transcripts via ataxia telangiectasia mutated and ataxia telangiectasia and Rad3 related (ATM-ATR) pathway, and suggested that radiation therapy might be used to increase the translation of NKG2D ligands. Two NSCLC cell lines, that is, A549 and NCI-H23 cells, were used to investigate the combined effects of ionizing radiation and HDAC inhibitors on the expressions of five NKG2D ligands. The mRNA expressions of the NKG2D ligands were quantitated by multiplex reverse transcription-PCR. Surface protein expressions were measured by flow cytometry, and the susceptibilities of cancer cells to NK cells were assayed by time-resolved fluorometry using the DELFIA® EuTDA cytotoxicity kit and by flow cytometry. The expressions of NKG2D ligands were found to be regulated at the transcription and translation levels. Ionizing radiation and HDAC inhibitors in combination synergistically increased the expressions of NKG2D ligands. Furthermore, treatment with ATM-ATR inhibitors efficiently blocked the increased translations of NKG2D ligands induced by ionizing radiation but did not block the increased ligand translations induced by HDAC inhibitors. The study confirms that increased NKG

  2. Analysis of the Interactions of Botanical Extract Combinations Against the Viability of Prostate Cancer Cell Lines

    OpenAIRE

    Adams, Lynn S.; Seeram, Navindra P.; Hardy, Mary L.; Catherine Carpenter; David Heber

    2006-01-01

    Herbal medicines are often combinations of botanical extracts that are assumed to have additive or synergistic effects. The purpose of this investigation was to compare the effect of individual botanical extracts with combinations of extracts on prostate cell viability. We then modeled the interactions between botanical extracts in combination isobolographically. Scutellaria baicalensis, Rabdosia rubescens, Panax-pseudo ginseng, Dendranthema morifolium, Glycyrrhiza uralensis and Serenoa re...

  3. Mechanotransduction Across the Cell Surface and Through the Cytoskeleton

    Science.gov (United States)

    Wang, Ning; Butler, James P.; Ingber, Donald E.

    1993-05-01

    Mechanical stresses were applied directly to cell surface receptors with a magnetic twisting device. The extracellular matrix receptor, integrin β_1, induced focal adhesion formation and supported a force-dependent stiffening response, whereas nonadhesion receptors did not. The cytoskeletal stiffness (ratio of stress to strain) increased in direct proportion to the applied stress and required intact microtubules and intermediate filaments as well as microfilaments. Tensegrity models that incorporate mechanically interdependent struts and strings that reorient globally in response to a localized stress mimicked this response. These results suggest that integrins act as mechanoreceptors and transmit mechanical signals to the cytoskeleton. Mechanotransduction, in turn, may be mediated simultaneously at multiple locations inside the cell through force-induced rearrangements within a tensionally integrated cytoskeleton.

  4. Surface plasmon enhanced cell microscopy with blocked random spatial activation

    Science.gov (United States)

    Son, Taehwang; Oh, Youngjin; Lee, Wonju; Yang, Heejin; Kim, Donghyun

    2016-03-01

    We present surface plasmon enhanced fluorescence microscopy with random spatial sampling using patterned block of silver nanoislands. Rigorous coupled wave analysis was performed to confirm near-field localization on nanoislands. Random nanoislands were fabricated in silver by temperature annealing. By analyzing random near-field distribution, average size of localized fields was found to be on the order of 135 nm. Randomly localized near-fields were used to spatially sample F-actin of J774 cells (mouse macrophage cell-line). Image deconvolution algorithm based on linear imaging theory was established for stochastic estimation of fluorescent molecular distribution. The alignment between near-field distribution and raw image was performed by the patterned block. The achieved resolution is dependent upon factors including the size of localized fields and estimated to be 100-150 nm.

  5. Remote Control of Tissue Interactions via Engineered Photo-switchable Cell Surfaces

    OpenAIRE

    Wei Luo; Abigail Pulsipher; Debjit Dutta; Lamb, Brian M.; Yousaf, Muhammad N.

    2014-01-01

    We report a general cell surface molecular engineering strategy via liposome fusion delivery to create a dual photo-active and bio-orthogonal cell surface for remote controlled spatial and temporal manipulation of microtissue assembly and disassembly. Cell surface tailoring of chemoselective functional groups was achieved by a liposome fusion delivery method and quantified by flow cytometry and characterized by a new cell surface lipid pull down mass spectrometry strategy. Dynamic co-culture ...

  6. Combined MET inhibition and topoisomerase I inhibition block cell growth of small cell lung cancer.

    Science.gov (United States)

    Rolle, Cleo E; Kanteti, Rajani; Surati, Mosmi; Nandi, Suvobroto; Dhanasingh, Immanuel; Yala, Soheil; Tretiakova, Maria; Arif, Qudsia; Hembrough, Todd; Brand, Toni M; Wheeler, Deric L; Husain, Aliya N; Vokes, Everett E; Bharti, Ajit; Salgia, Ravi

    2014-03-01

    Small cell lung cancer (SCLC) is a devastating disease, and current therapies have not greatly improved the 5-year survival rates. Topoisomerase (Top) inhibition is a treatment modality for SCLC; however, the response is short lived. Consequently, our research has focused on improving SCLC therapeutics through the identification of novel targets. Previously, we identified MNNG HOS transforming gene (MET) to be overexpressed and functional in SCLC. Herein, we investigated the therapeutic potential of combinatorial targeting of MET using SU11274 and Top1 using 7-ethyl-10-hydroxycamptothecin (SN-38). MET and TOP1 gene copy numbers and protein expression were determined in 29 patients with limited (n = 11) and extensive (n = 18) disease. MET gene copy number was significantly increased (>6 copies) in extensive disease compared with limited disease (P = 0.015). Similar TOP1 gene copy numbers were detected in limited and extensive disease. Immunohistochemical staining revealed a significantly higher Top1 nuclear expression in extensive (0.93) versus limited (0.15) disease (P = 0.04). Interestingly, a significant positive correlation was detected between MET gene copy number and Top1 nuclear expression (r = 0.5). In vitro stimulation of H82 cells revealed hepatocyte growth factor (HGF)-induced nuclear colocalization of p-MET and Top1. Furthermore, activation of the HGF/MET axis enhanced Top1 activity, which was abrogated by SU11274. Combination of SN-38 with SU11274 dramatically decreased SCLC growth as compared with either drug alone. Collectively, these findings suggest that the combinatorial inhibition of MET and Top1 is a potentially efficacious treatment strategy for SCLC. PMID:24327519

  7. Surface-engineered substrates for improved human pluripotent stem cell culture under fully defined conditions

    OpenAIRE

    Saha, Krishanu; Mei, Ying; Reisterer, Colin M.; Pyzocha, Neena Kenton; Yang, Jing; Muffat, Julien; Davies, Martyn C.; Alexander, Morgan R.; Langer, Robert; Anderson, Daniel G.; Jaenisch, Rudolf

    2011-01-01

    The current gold standard for the culture of human pluripotent stem cells requires the use of a feeder layer of cells. Here, we develop a spatially defined culture system based on UV/ozone radiation modification of typical cell culture plastics to define a favorable surface environment for human pluripotent stem cell culture. Chemical and geometrical optimization of the surfaces enables control of early cell aggregation from fully dissociated cells, as predicted from a numerical model of cell...

  8. Surface Properties of Cell-treated Polyethylene Terephthalate

    Directory of Open Access Journals (Sweden)

    Bing Shi

    2006-01-01

    Full Text Available The materials used in artificial joints undergo degradation through fatigue and corrosive wear in human body. The lifetime for well-designed artificial joints like hip joints is at most 12 years and a patient will usually have two total joint replacements during his/her lifetime. Tissue engineering, an alternative to total joint implantation, is the replacement of damaged tissue with the tissue that is designed and constructed to meet the needs of the individual patient. In this study, polyethylene terephthalate (PET in the form of overhead transparency films were investigated on their cell interactions and the tribological properties as an alternative tissue-engineering matrix. The base material of the transparency films is PET. Cell culture methods as well as atomic force microscope (AFM, contact angle goniometer, confocal microscope and universal tribotester were used to study the properties of the substrate materials and the interactions between the surface and the substrate materials. Results showed that cells grew on the substrate of the base materials of the PET. The tribological properties of the slides have been changed after being cell-treated.

  9. Apoptosis of human gastric cancer SGC-7901 cells induced by mitomycin combined with sulindac

    Institute of Scientific and Technical Information of China (English)

    Li Ma; Yong-Le Xie; Yi Yu; Qiu-Ning Zhang

    2005-01-01

    AIM: To investigate the effects of mitomycin (MMC)combined with sulindac on cell viability, apoptotic induction and expression of apoptosis-related gene Bcl-2 and cyclooxygenase-2 (COX-2)in gastric cancer SGC-7901cells.METHODS: Human gastric cancer SGC-7901 cells were divided into three treatment groups,namely sulindac treatment group, MMC treatment group and combined sulindac with MMC treatment group. After being treated with drugs, cell viability was examined by MTr assay.Flow cytometry was used to evaluate the cell cycle distribution and apoptotic rates. Morphology of the cells was observed under light microscope and interactive laser microscope. Expression of COX-2 and Bcl-2 was determined by immunocytochemical method.RESULTS: After exposure for 12 h to three kinds of drugs,gastric cancer SGC-7901 cells presented some morphological features of apoptosis, including cell shrinkage, nuclear condensation, DNA fragmentation and formation of apoptotic bodies. Growth inhibition was more obvious in combined sulindac with MMC treatment group and sulindac treatment group than in MMC treatment group. The apoptotic rates in co-treated cells and MMC-treated cells 24 h after treatment were 12.0% and 7.2%, respectively.After exposure for 24 h to MMC, the expression of COX-2and Bcl-2 protein was up-regulated, COX-2 levels were down-regulated but Bcl-2 gene expression was not changed significantly in combined treatment group.CONCLUSION: MMC-induced apoptosis is reduced by up-regulating the expression of COX-2 and Bcl-2 genes.MMC combined with sulindac can suppress the growth of gastric cancer cells through induction of apoptosis mediated by down-regulation of apoptosis-related Bcl-2and COX-2 gene.

  10. Combined Cell Culture-Biosensing Platform Using Vertically Aligned Patterned Peptide Nanofibers for Cellular Studies

    DEFF Research Database (Denmark)

    Taskin, Mehmet B.; Sasso, Luigi; Dimaki, Maria;

    2013-01-01

    This Article presents the development of a combined cell culture–biosensing platform using vertically aligned self-assembled peptide nanofibers. Peptide nanofibers were patterned on a microchip containing gold microelectrodes to provide the cells with a 3D environment enabling them to grow...

  11. Probing the compressibility of tumor cell nuclei by combined atomic force-confocal microscopy

    NARCIS (Netherlands)

    Krause, M.; Riet, J. te; Wolf, K. van der

    2013-01-01

    The cell nucleus is the largest and stiffest organelle rendering it the limiting compartment during migration of invasive tumor cells through dense connective tissue. We here describe a combined atomic force microscopy (AFM)-confocal microscopy approach for measurement of bulk nuclear stiffness toge

  12. Curcumin combined FOLFOX induced cell apoptosis of gastric cancer and its mechanism

    Institute of Scientific and Technical Information of China (English)

    周香

    2013-01-01

    Objective To observe the effect of curcumin combined folinic acid fluorouracil oxaliplatin (FOLFOX) on the gastric adenocarcinoma cell line BGC-823 and to explore its possible mechanisms.Methods Cells were divided into five groups,i.e.the blank control group,the

  13. Tnhibitory effect of Fuzheng Yiliuyin in combination with chemotherapeutics on human gastric carcinoma cell strain

    Institute of Scientific and Technical Information of China (English)

    Yi Liu; Rui Wang; Gen-Quan Qiu; Ke-Jun Nan; Xi-Cai Sun

    2006-01-01

    AIM: To study the inhibitory effects of Fuzheng Yiliuyin (Decoction for Suppressing Tumors by Strengthening the Body Resistance) in combination with chemotherapeutics on human gastric carcinoma cell strain.METHODS: Fuzheng Yiliuyin (ZY) combined with various kinds of chemotherapeutics was put into two kinds of cultivated human gastric carcinoma cell strains,then its inhibitory effects on human gastric carcinoma cell strains were determind by the MTT method. Flow cytometer was used to assay the apoptosis rate, and the ultrastructure of gastric carcinoma cells was observed under transmission electron microscope.RESULTS: Obvious apoptosis was seen in gastric carcinoma cells after treatment with ZY for 72 h. ZY and chemical drugs had synergistic inhibition effects on the cultivated gastric carcinoma cells, but the effects were different on various cell strains. The inhibitory effects of ZY could be strengthened by cytotoxic action and apoptosis. ZY combined with fluorouracil, etoposide and cisplatin (EFP) chemotherapeutics had better inhibitory effects on SGC-7901, while ZY combined with EFP or with DDP chemotherapeutics had better inhibitory effects than other drugs on MGC-803.CONCLUSION: ZY induces apoptosis and inhibits the growth of gastric carcinoma cells. ZY has the synergistic function of chemotherapeutics.

  14. A combined surface and borehole seismic survey at the COSC-1 borehole

    Science.gov (United States)

    Simon, Helge; Krauß, Felix; Hedin, Peter; Buske, Stefan; Giese, Rüdiger; Juhlin, Christopher

    2015-04-01

    The ICDP project COSC (Collisional Orogeny in the Scandinavian Caledonides) focuses on the mid Paleozoic Caledonide Orogen in Scandinavia in order to better understand orogenic processes, from the past and in recent active mountain belts. The Scandinavian Caledonides provide a well preserved example of a Paleozoic continent-continent collision. Surface geology in combination with geophysical data provide control of the geometry of the Caledonian structure, including the allochthon and the underlying autochthon, as well as the shallow W-dipping décollement surface that separates the two and consist of a thin skin of Cambrian black shales. During spring/summer 2014 the COSC-1 borehole was drilled to approx. 2.5 km depth near the town of Åre (western Jämtland/Sweden) with nearly 100 % of core recovery and cores in best quality. After the drilling was finished, a major seismic survey was conducted in and around the COSC-1 borehole which comprised both seismic reflection and transmission experiments. Besides a high resolution zero-offset VSP (Vertical Seismic Profiling) experiment also a multi-azimuthal walkaway VSP survey took place. For the latter the source points were distributed along three profile lines centered radially around the borehole. For the central part up to 2.5 km away from the borehole, a hydraulic hammer source was used, which hits the ground for about 20 s with an linear increasing hit rate. For the far offset shots up to 5 km, explosive sources were used. The wavefield of both source types was recorded in the borehole using an array of 15 three-component receivers with a geophone spacing of 10 m. This array was deployed at 7 different depth levels during the survey. At the same time the wavefield was also recorded at the surface by 180 standalone three-component receivers placed along each of the three up to 10 km long lines, as well as with a 3D array of single-component receivers in the central part of the survey area around the borehole. Here

  15. Cell surface and transcriptional characterization of human adipose-derived adherent stromal (hADAS) cells.

    Science.gov (United States)

    Katz, Adam J; Tholpady, Ashok; Tholpady, Sunil S; Shang, Hulan; Ogle, Roy C

    2005-03-01

    Adult human subcutaneous adipose tissue contains cells with intriguing multilineage developmental plasticity, much like marrow-derived mesenchymal stem cells. Putative stem or progenitor cells from fat have been given many different names in the literature, reflecting an early and evolving consensus regarding their phenotypic characterization. The study reported here used microarrays to evaluate over 170 genes relating to angiogenesis and extracellular matrix in undifferentiated, early-passage human adipose-derived adherent stromal (hADAS) cells isolated from three separate donors. The hADAS populations unanimously transcribed 66% of the screened genes, and 83% were transcribed by at least two of the three populations. The most highly transcribed genes relate to functional groupings such as cell adhesion, matrix proteins, growth factors and receptors, and proteases. The transcriptome of hADAS cells demonstrated by this work reveals many similarities to published profiles of bone marrow mesenchymal stem cells (MSCs). In addition, flow analysis of over 24 hADAS cell surface proteins (n = 7 donors) both confirms and expands on the existing literature and reveals strong intergroup correlation, despite an inconsistent nomenclature and the lack of standardized protocols for cell isolation and culture. Finally, based on flow analysis and reverse transcription polymerase chain reaction studies, our results suggest that hADAS cells do not express several proteins that are implicated as markers of "stemness" in other stem cell populations, including telomerase, CD133, and the membrane transporter ABCG2.

  16. IL-4 enhances expression and function of surface IgM in CLL cells.

    Science.gov (United States)

    Aguilar-Hernandez, Maria M; Blunt, Matthew D; Dobson, Rachel; Yeomans, Alison; Thirdborough, Stephen; Larrayoz, Marta; Smith, Lindsay D; Linley, Adam; Strefford, Jonathan C; Davies, Andrew; Johnson, Peter M W; Savelyeva, Natalia; Cragg, Mark S; Forconi, Francesco; Packham, Graham; Stevenson, Freda K; Steele, Andrew J

    2016-06-16

    Kinase inhibitors targeting the B-cell receptor (BCR) are now prominent in the treatment of chronic lymphocytic leukemia (CLL). We have focused here on interleukin 4 (IL-4), a cytokine that protects normal and malignant B cells from apoptosis and increases surface immunoglobulin M (sIgM) expression on murine splenic B cells. First, we have demonstrated that IL-4 treatment increased sIgM expression in vitro on peripheral blood B cells obtained from healthy individuals. In CLL, IL-4 target genes are overexpressed in cells purified from the lymph nodes of patients compared with cells derived from matched blood and bone marrow samples. As for normal B cells, IL-4 increased sIgM expression on CLL cells in vitro, especially in samples expressing unmutated V-genes. IL-4-induced sIgM expression was associated with increased receptor signalling activity, measured by anti-IgM-induced calcium mobilization, and with increased expression of CD79B messenger RNA and protein, and the "mature" glycoform of sIgM. Importantly, the ability of the BCR-associated kinase inhibitors idelalisib and ibrutinib, approved for treatment of CLL and other B-cell malignancies, to inhibit anti-IgM-induced signalling was reduced following IL-4 pretreatment in samples from the majority of patients. In contrast to stimulatory effects on sIgM, IL-4 decreased CXCR4 and CXCR5 expression; therefore, CLL cells, particularly within the progressive unmutated V-gene subset, may harness the ability of IL-4 to promote BCR signalling and B-cell retention within lymph nodes. Effects of IL-4 were mediated via JAK3/STAT6 and we propose a potential role for JAK inhibitors in combination with BCR kinase inhibitors for the treatment of CLL. PMID:27002119

  17. Electrospun fibrous scaffolds combined with nanoscale hydroxyapatite induce osteogenic differentiation of human periodontal ligament cells.

    Science.gov (United States)

    Wu, Xiaonan; Miao, Leiying; Yao, Yingfang; Wu, Wenlei; Liu, Yu; Chen, Xiaofeng; Sun, Weibin

    2014-01-01

    Periodontal repair is a complex process in which regeneration of alveolar bone is a vital component. The aim of this study was to develop a biodegradable scaffold with good biocompatibility and osteoinductive ability. Two types of composite fibrous scaffolds were produced by electrospinning, ie, type I collagen/poly(ε-caprolactone) (COL/PCL) and type I collagen/poly(ε-caprolactone)/nanoscale hydroxyapatite (COL/PCL/nHA) with an average fiber diameter of about 377 nm. After a simulated body fluid (SBF) immersion test, the COL/PCL/nHA-SBF scaffold developed a rough surface because of the calcium phosphate deposited on the fibers, suggesting that the presence of nHA promoted the mineralization potential of the scaffold. Energy dispersive X-ray spectroscopy clearly showed the calcium and phosphorus content in the COL/PCL/nHA and COL/PCL/nHA-SBF scaffolds, confirming the findings of nHA and calcium phosphate precipitation on scanning electron micrographs. Water contact analysis revealed that nHA could improve the hydrophilic nature of the COL/PCL/nHA-SBF scaffold. The morphology of periodontal ligament cells cultured on COL/PCL-SBF and COL/PCL/nHA-SBF was evaluated by scanning electron microscopy. The results showed that cells adhered to either type of scaffold and were slightly spindle-shaped in the beginning, then extended gradually with stretched filopodia, indicating an ability to fill the fiber pores. A Cell Counting Kit-8 assay showed that both scaffolds supported cell proliferation. However, real-time quantitative polymerase chain reaction analysis showed that expression of the bone-related markers, alkaline phosphatase and osteocalcin, was upregulated only on the COL/PCL/nHA-SBF scaffold, indicating that this scaffold had the ability to induce osteogenic differentiation of periodontal ligament cells. In this study, COL/PCL/nHA-SBF produced by electrospinning followed by biomimetic mineralization had combined electrospun fibers with nHA in it. This scaffold has

  18. Combining 3D human in vitro methods for a 3Rs evaluation of novel titanium surfaces in orthopaedic applications

    Science.gov (United States)

    Stevenson, G.; Rehman, S.; Draper, E.; Hernández‐Nava, E.; Hunt, J.

    2016-01-01

    ABSTRACT In this study, we report on a group of complementary human osteoblast in vitro test methods for the preclinical evaluation of 3D porous titanium surfaces. The surfaces were prepared by additive manufacturing (electron beam melting [EBM]) and plasma spraying, allowing the creation of complex lattice surface geometries. Physical properties of the surfaces were characterized by SEM and profilometry and 3D in vitro cell culture using human osteoblasts. Primary human osteoblast cells were found to elicit greater differences between titanium sample surfaces than an MG63 osteoblast‐like cell line, particularly in terms of cell survival. Surface morphology was associated with higher osteoblast metabolic activity and mineralization on rougher titanium plasma spray coated surfaces than smoother surfaces. Differences in osteoblast survival and metabolic activity on titanium lattice structures were also found, despite analogous surface morphology at the cellular level. 3D confocal microscopy identified osteoblast organization within complex titanium surface geometries, adhesion, spreading, and alignment to the biomaterial strut geometries. Mineralized nodule formation throughout the lattice structures was also observed, and indicative of early markers of bone in‐growth on such materials. Testing methods such as those presented are not traditionally considered by medical device manufacturers, but we suggest have value as an increasingly vital tool in efficiently translating pre‐clinical studies, especially in balance with current regulatory practice, commercial demands, the 3Rs, and the relative merits of in vitro and in vivo studies. Biotechnol. Bioeng. 2016;113: 1586–1599. © 2015 The Authors. Biotechnology and Bioengineering Published by Wiley Periodicals, Inc. PMID:26702609

  19. In vitro evaluation of photon and carbon ion radiotherapy in combination with chemotherapy in glioblastoma cells

    Directory of Open Access Journals (Sweden)

    Combs Stephanie E

    2012-01-01

    Full Text Available Abstract Background To evaluate the cytotoxic effect of carbon ion radiotherapy and chemotherapy in glioblastoma cells in vitro. Methods and Materials The human glioblastoma (GBM cell line U87 was irradiated with photon radiotherapy (RT doses of 2 Gy, 4 Gy and 6 Gy. Likewise, irradiation with carbon ions was performed with single carbon doses of 0.125, 0.5, 2 and 3 Gy. Four chemotherapeutic substances, camptothecin, gemcitabine, paclitaxel and cisplatinum, were used for single and combination experiments. The assessment of the effect of single and double treatment on cell viability was performed using the clonogenic growth assay representing the radiobiological gold standard. Results The RBE of carbon ions ranges between 3.3 and 3.9 depending on survival level and dose. All chemotherapeutic substances showed a clear does-response relationhips. in their characteristic concentrations. For subsequent combination experiments, two dose levels leading to low and medium reduction of cell survival were chosen. Combination experiments showed additive effects independently of the drugs' mechanisms of action. Paclitaxel and campthothecin demonstrated the most prominent cytotoxic effect in combination with carbon ion radiotherapy. Conclusion In conclusion, combination of carbon ion radiotherapy with chemotherapies of different mechanisms of action demonstrates additive effects. The most dominant effect was produced by paclitaxel, followed by camptothecin, as espected from previously published work. The present data serve as an important radiobiological basis for further combination experiments, as well as clinical studies on combination treatments.

  20. The combinational effect of vincristine and berberine on growth inhibition and apoptosis induction in hepatoma cells.

    Science.gov (United States)

    Wang, Ling; Wei, Dandan; Han, Xiaojuan; Zhang, Wei; Fan, Chengzhong; Zhang, Jie; Mo, Chunfen; Yang, Ming; Li, Junhong; Wang, Zhe; Zhou, Qin; Xiao, Hengyi

    2014-04-01

    The use of vincristine, a known antitumor agent, in hepatoma therapy is limited particularly because of its toxic effect. Meanwhile, berberine has drawn increasing attention to its antineoplastic effect in recent years. In view of the advantages of combinational drug treatment reported in anti-cancer chemotherapy, we evaluated the effects of co-treatment of vincristine and berberine on hepatic carcinoma cell lines in this study. We find that combinational usage of these two drugs can significantly induce cell growth inhibition and apoptosis even under a concentration of vincristine barely showing cytotoxicity in the same cells when used alone. The underlying mechanism about this combinational effect was addressed in this study by monitoring the signals related to mitochondrial function, apoptotic pathway and endoplasmic reticulum stress. Our results suggest a new value of berberine as a potential adjuvant agent in cancer chemotherapy and provide a hopeful approach for developing hepatoma therapy by utilizing the combinational effect of vincristine and berberine.

  1. Effects of Celecoxib and Ly117018 Combination on Human Breast Cancer Cells in Vitro

    Directory of Open Access Journals (Sweden)

    Klaus H. Baumann

    2009-01-01

    Full Text Available Activation and signalling of estrogen receptor (ER and COX-2 represent two important pathways in breast cancer cell regulation. Activation of either pathway is associated with breast cancer cell proliferation and eventually malignant progression. Raloxifene analogue, Ly117018, a selective estrogen receptor modulator and celecoxib, a specific COX- 2 inhibitor have been shown to inhibit breast cancer cell proliferation when used alone in vitro and in vivo. In this study, the combined drug effects on hormone-dependent MCF-7 and hormone-independent MDA-MB-435 cells in vitro were evaluated. Cell proliferation assays excluded drug antagonism and revealed a moderate synergistic growth inhibitory activity of Ly117018 and celecoxib on both cell lines when combined in specific concentrations. Growth inhibition of either compound was not associated with cell cycle arrest. In MCF-7 cells, western blot analysis revealed a decreased phosphorylation of the AKT protein by either agent alone or in combination. In MDA-MB-435 cells, celecoxib alone induced an increase in AKT phosphorylation relative to total AKT protein; this effect was decreased in the presence of Ly117018. These results indicate that these two drugs are non-antagonistic; and when combined in specific concentrations, moderate synergistic antiproliferative activity of celecoxib and Ly117018 were observed in hormone-dependent MCF-7 and hormone- independent MDA-MB-435 cells associated with changes in cell cycle distribution and regulation of AKT protein and phosphorylation. These findings further support a central role of the ER- and COX-2 pathways in human breast cancer cells.

  2. A Model Combining Oscillations and Attractor Dynamics for Generation of Grid Cell Firing

    OpenAIRE

    Michael E Hasselmo; Brandon, Mark P.

    2012-01-01

    Different models have been able to account for different features of the data on grid cell firing properties, including the relationship of grid cells to cellular properties and network oscillations. This paper describes a model that combines elements of two major classes of models of grid cells: models using interference of oscillations and models using attractor dynamics. This model includes a population of units with oscillatory input representing input from the medial septum. These units ...

  3. Combining antiangiogenic therapy with adoptive cell immunotherapy exerts better antitumor effects in non-small cell lung cancer models.

    Directory of Open Access Journals (Sweden)

    Shujing Shi

    Full Text Available INTRODUCTION: Cytokine-induced killer cells (CIK cells are a heterogeneous subset of ex-vivo expanded T lymphocytes which are characterized with a MHC-unrestricted tumor-killing activity and a mixed T-NK phenotype. Adoptive CIK cells transfer, one of the adoptive immunotherapy represents a promising nontoxic anticancer therapy. However, in clinical studies, the therapeutic activity of adoptive CIK cells transfer is not as efficient as anticipated. Possible explanations are that abnormal tumor vasculature and hypoxic tumor microenvironment could impede the infiltration and efficacy of lymphocytes. We hypothesized that antiangiogenesis therapy could improve the antitumor activity of CIK cells by normalizing tumor vasculature and modulating hypoxic tumor microenvironment. METHODS: We combined recombinant human endostatin (rh-endostatin and CIK cells in the treatment of lung carcinoma murine models. Intravital microscopy, dynamic contrast enhanced magnetic resonance imaging, immunohistochemistry, and flow cytometry were used to investigate the tumor vasculature and hypoxic microenvironment as well as the infiltration of immune cells. RESULTS: Our results indicated that rh-endostatin synergized with adoptive CIK cells transfer to inhibit the growth of lung carcinoma. We found that rh-endostatin normalized tumor vasculature and reduced hypoxic area in the tumor microenvironment. Hypoxia significantly inhibited the proliferation, cytotoxicity and migration of CIK cells in vitro and impeded the homing of CIK cells into tumor parenchyma ex vivo. Furthermore, we found that treatment with rh-endostatin significantly increased the homing of CIK cells and decreased the accumulation of suppressive immune cells in the tumor tissue. In addition, combination therapy produced higher level of tumor-infiltration lymphocytes compared with other treatments. CONCLUSIONS: Our results demonstrate that rh-endostatin improves the therapeutic effect of adoptive CIK cells

  4. Combining Antiangiogenic Therapy with Adoptive Cell Immunotherapy Exerts Better Antitumor Effects in Non-Small Cell Lung Cancer Models

    Science.gov (United States)

    Shi, Shujing; Wang, Rui; Chen, Yitian; Song, Haizhu; Chen, Longbang; Huang, Guichun

    2013-01-01

    Introduction Cytokine-induced killer cells (CIK cells) are a heterogeneous subset of ex-vivo expanded T lymphocytes which are characterized with a MHC-unrestricted tumor-killing activity and a mixed T-NK phenotype. Adoptive CIK cells transfer, one of the adoptive immunotherapy represents a promising nontoxic anticancer therapy. However, in clinical studies, the therapeutic activity of adoptive CIK cells transfer is not as efficient as anticipated. Possible explanations are that abnormal tumor vasculature and hypoxic tumor microenvironment could impede the infiltration and efficacy of lymphocytes. We hypothesized that antiangiogenesis therapy could improve the antitumor activity of CIK cells by normalizing tumor vasculature and modulating hypoxic tumor microenvironment. Methods We combined recombinant human endostatin (rh-endostatin) and CIK cells in the treatment of lung carcinoma murine models. Intravital microscopy, dynamic contrast enhanced magnetic resonance imaging, immunohistochemistry, and flow cytometry were used to investigate the tumor vasculature and hypoxic microenvironment as well as the infiltration of immune cells. Results Our results indicated that rh-endostatin synergized with adoptive CIK cells transfer to inhibit the growth of lung carcinoma. We found that rh-endostatin normalized tumor vasculature and reduced hypoxic area in the tumor microenvironment. Hypoxia significantly inhibited the proliferation, cytotoxicity and migration of CIK cells in vitro and impeded the homing of CIK cells into tumor parenchyma ex vivo. Furthermore, we found that treatment with rh-endostatin significantly increased the homing of CIK cells and decreased the accumulation of suppressive immune cells in the tumor tissue. In addition, combination therapy produced higher level of tumor-infiltration lymphocytes compared with other treatments. Conclusions Our results demonstrate that rh-endostatin improves the therapeutic effect of adoptive CIK cells therapy against lung

  5. Combined Use of Metformin and Everolimus Is Synergistic in the Treatment of Breast Cancer Cells.

    Science.gov (United States)

    Wang, Yunshan; Wei, Junmin; Li, Li; Fan, Cong; Sun, Ying

    2014-01-01

    Everolimus inhibits mammalian target of rapamycin (mTOR) and leads to decreased protein synthesis and decreased cancer cell proliferation in many experimental systems. Adenosine 5'-monophosphate-activated protein kinase (AMPK) activators such as metformin have similar actions in keeping with the TSC2/1 pathway linking activation of AMPK to inhibition of mTOR. Histopathological and biochemical studies of breast cancer show frequent dysregulation of the AMPK and the mTOR pathway. Therefore, we investigated the efficacy of the mTOR inhibitor everolimus and metformin in the treatment of breast cancer cells. This study evaluated the in vitro and in vivo effects of everolimus alone or in combination with metformin on breast cancer cells. MTT assay was used to quantify the inhibitory effect of the drugs on breast cancer cells in vitro. SCID mice injected with HCC1428 cells followed by different treatments were used to assess the in vivo efficacy of different agents. Data showed that the combination of everolimus and metformin exerted synergistic inhibitory effects on the growth of breast cancer cells both in culture and in a mouse xenograft model. Further, this combination abrogated S6 and 4EBP1phosphorylation. Collectively, we suggest that the combination of everolimus and metformin may be an effective regimen for treatment of breast cancer, hence warranting further evaluation of the combination in the clinic. PMID:26351208

  6. A model combining oscillations and attractor dynamics for generation of grid cell firing

    Directory of Open Access Journals (Sweden)

    Michael E Hasselmo

    2012-05-01

    Full Text Available Different models have been able to account for different features of the data on grid cell firing properties, including the relationship of grid cells to cellular properties and network oscillations. This paper describes a model that combines elements of two major classes of models of grid cells: models using interference of oscillations and models using attractor dynamics. This model includes a population of units with oscillatory input representing input from the medial septum. These units are termed heading angle cells because their connectivity depends upon heading angle in the environment as well as the spatial phase coded by the cell. These cells project to a population of grid cells. The sum of the heading angle input results in standing waves of circularly symmetric input to the grid cell population. Feedback from the grid cell population increases the activity of subsets of the heading angle cells, resulting in the network settling into activity patterns that resemble the patterns of firing fields in a population of grid cells. The properties of heading angle cells firing as conjunctive grid-by-head-direction cells can shift the grid cell firing according to movement velocity. The pattern of interaction of oscillations requires use of separate populations that fire on alternate cycles of the net theta rhythmic input to grid cells, similar to recent neurophysiological data on theta cycle skipping in medial entorhinal cortex.

  7. Facile modification of gelatin-based microcarriers with multiporous surface and proliferative growth factors delivery to enhance cell growth

    International Nuclear Information System (INIS)

    The design of microcarriers plays an important role in the success of cell expansion. The present article provides a facile approach to modify the gelatin-based particles and investigates the feasibility of their acting as microcarriers for cell attachment and growth. Gelatin particles (150-320 μm) were modified by cryogenic treatment and lyophilization to develop the surface with the features of multiporous morphology and were incorporated with proliferative growth factors (bFGF) by adsorption during the post-preparation, which enables them to serve as microcarriers for cells amplification, together with the advantages of larger cell-surface contact area and capability of promoting cell propagation. The microstructure and release assay of the modified microcarriers demonstrated that the pores on surface were uniform and bFGF was released in a controlled manner. Through in vitro fibroblast culture, these features resulted in a prominent increase in the cell attachment rate and cell growth rate relative to the conditions without modification. Although the scanning electron microscopy and optical microscopy analysis results indicated that cells attached, spread, and proliferated on all the microcarriers, cell growth clearly showed a significant correlation with the multiporous structure of microcarriers, in particular on bFGF combined ones. These results validate our previous assumption that the facile modification could improve cell growth on the gelatin-based microcarriers obviously and the novel microcarriers may be a promising candidate in tissue engineering

  8. Facile modification of gelatin-based microcarriers with multiporous surface and proliferative growth factors delivery to enhance cell growth

    Energy Technology Data Exchange (ETDEWEB)

    Huang Sha [Department of Oral Histology and Pathology, School of Stomatology, Fourth Military Medical University, Xi' an 710032 (China); Research and Development Center for Tissue Engineering, Fourth Military Medical University, Xi' an 710032 (China); Wang Yijuan [Key Laboratory for Macromolecular Science of Shaanxi Province, Shaanxi Normal University, Xi' an 710062 (China); Deng, Tianzheng [Research and Development Center for Tissue Engineering, Fourth Military Medical University, Xi' an 710032 (China); Department of Oral and Maxillofacial Surgery, School of Stomatology, Fourth Military Medical University, Xi' an 710032 (China); Jin Fang [Department of Orthodontics, School of Stomatology, Fourth Military Medical University, Xi' an, 710032 (China); Liu Shouxin [Key Laboratory for Macromolecular Science of Shaanxi Province, Shaanxi Normal University, Xi' an 710062 (China); Zhang Yongjie [Department of Oral Histology and Pathology, School of Stomatology, Fourth Military Medical University, Xi' an 710032 (China); Research and Development Center for Tissue Engineering, Fourth Military Medical University, Xi' an 710032 (China); Feng Feng [Research and Development Center for Tissue Engineering, Fourth Military Medical University, Xi' an 710032 (China); Department of Dermatology, Tangdu Hospital, Fourth Military Medical University, Xi' an 710038 (China); Jin Yan [Department of Oral Histology and Pathology, School of Stomatology, Fourth Military Medical University, Xi' an 710032 (China); Research and Development Center for Tissue Engineering, Fourth Military Medical University, Xi' an 710032 (China)], E-mail: yanjin@fmmu.edu.cn

    2008-07-28

    The design of microcarriers plays an important role in the success of cell expansion. The present article provides a facile approach to modify the gelatin-based particles and investigates the feasibility of their acting as microcarriers for cell attachment and growth. Gelatin particles (150-320 {mu}m) were modified by cryogenic treatment and lyophilization to develop the surface with the features of multiporous morphology and were incorporated with proliferative growth factors (bFGF) by adsorption during the post-preparation, which enables them to serve as microcarriers for cells amplification, together with the advantages of larger cell-surface contact area and capability of promoting cell propagation. The microstructure and release assay of the modified microcarriers demonstrated that the pores on surface were uniform and bFGF was released in a controlled manner. Through in vitro fibroblast culture, these features resulted in a prominent increase in the cell attachment rate and cell growth rate relative to the conditions without modification. Although the scanning electron microscopy and optical microscopy analysis results indicated that cells attached, spread, and proliferated on all the microcarriers, cell growth clearly showed a significant correlation with the multiporous structure of microcarriers, in particular on bFGF combined ones. These results validate our previous assumption that the facile modification could improve cell growth on the gelatin-based microcarriers obviously and the novel microcarriers may be a promising candidate in tissue engineering.

  9. Association of cell surface heparan sulfate proteoglycans of Schwann cells with extracellular matrix proteins.

    Science.gov (United States)

    Carey, D J; Crumbling, D M; Stahl, R C; Evans, D M

    1990-11-25

    The terminal differentiation of Schwann cells is dependent on contact with basement membrane. The present study was undertaken to investigate the role of cell surface heparan sulfate proteoglycans (HSPGs) in mediating Schwann cell responses to extracellular matrix contact. Phosphatidylinositol-specific phospholipase C-releasable cell surface HSPGs purified from cultures of neonatal rat Schwann cells were subjected to affinity chromatography on immobilized laminin and fibronectin. Binding of the HSPG to both affinity matrices was observed. The strength of the association, however, was sensitive to the ionic strength of the buffer. In 0.1 M Tris-HCl, HSPG binding was essentially irreversible whereas in physiological ionic strength buffer (e.g. 0.142 M NaCl, 10 mM Tris), weaker binding was detected as a delay in elution of the HSPG from the affinity columns. Further studies of HSPG-laminin binding suggested that the binding was mediated by the glycosaminoglycan chains of the proteoglycans. Results of equilibrium gel filtration chromatography provided additional evidence for a reversible association of the HSPG and laminin with a Kd of approximately 1 x 10(-6) M. When Schwann cells were plated on plastic dishes coated with laminin, the cells attached and extended long slender processes. Inclusion of heparin, but not chondroitin sulfate, in the assay medium resulted in partial inhibition of process extension, but at concentrations of heparin which were higher than that needed to disrupt laminin-HSPG association in vitro. Addition of anti-integrin receptor antibodies resulted in more extensive inhibition of laminin-dependent process extension. Anti-integrin antibodies plus heparin essentially totally inhibited laminin-dependent process extension. These results demonstrate that cell surface HSPGs are capable of reversible association with extracellular matrix molecules and suggest that HSPG-laminin interactions play a role in laminin-dependent Schwann cell spreading. PMID

  10. Combined effects of C225 and 125-iodine seed radiation on colorectal cancer cells

    International Nuclear Information System (INIS)

    To characterize the effect of combined treatment of the anti-epidermal growth factor receptor (EGFR) monoclonal antibody C225 and 125-iodine (125I) seed radiation in human colorectal cancer. We treated LS180 cells with 125I continuous low dose rate radiation in the presence and absence of 100 nM C225. The clonogenic capacity, cellular proliferation, cell cycle distribution, apoptosis, and molecular pathways of the cells following the treatments were analyzed in vitro. The sensitizer enhancement ratio of C225 was approximately 1.4. Treatment with C225 and radiation alone produced significant inhibition of cell growth, but combination therapy produced greater inhibition than either treatment administered alone. C225 increased the radiation-induced apoptosis and the fraction of γ-H2AX foci positive cells at 48 h after treatment. The Akt phosphorylation level was lower in the cells receiving the combination treatment than in the cells treated with radiation or C225 alone. These findings indicate that C225 sensitizes LS180 cells to 125I seed radiation. Growth inhibition is mediated by inducing apoptosis and not cell cycle arrest. Additionally, we confirmed that C225 impairs DNA repair by reducing the cellular level of the DNA-PKcs and Ku70 proteins. Furthermore, the inhibition of Akt signaling activation may be responsible for the C225-mediated radiosensitization

  11. Cell surface area and membrane folding in glioblastoma cell lines differing in PTEN and p53 status.

    Directory of Open Access Journals (Sweden)

    Simon Memmel

    Full Text Available Glioblastoma multiforme (GBM is characterized by rapid growth, invasion and resistance to chemo-/radiotherapy. The complex cell surface morphology with abundant membrane folds, microvilli, filopodia and other membrane extensions is believed to contribute to the highly invasive behavior and therapy resistance of GBM cells. The present study addresses the mechanisms leading to the excessive cell membrane area in five GBM lines differing in mutational status for PTEN and p53. In addition to scanning electron microscopy (SEM, the membrane area and folding were quantified by dielectric measurements of membrane capacitance using the single-cell electrorotation (ROT technique. The osmotic stability and volume regulation of GBM cells were analyzed by video microscopy. The expression of PTEN, p53, mTOR and several other marker proteins involved in cell growth and membrane synthesis were examined by Western blotting. The combined SEM, ROT and osmotic data provided independent lines of evidence for a large variability in membrane area and folding among tested GBM lines. Thus, DK-MG cells (wild type p53 and wild type PTEN exhibited the lowest degree of membrane folding, probed by the area-specific capacitance C m = 1.9 µF/cm(2. In contrast, cell lines carrying mutations in both p53 and PTEN (U373-MG and SNB19 showed the highest C m values of 3.7-4.0 µF/cm(2, which corroborate well with their heavily villated cell surface revealed by SEM. Since PTEN and p53 are well-known inhibitors of mTOR, the increased membrane area/folding in mutant GBM lines may be related to the enhanced protein and lipid synthesis due to a deregulation of the mTOR-dependent downstream signaling pathway. Given that membrane folds and extensions are implicated in tumor cell motility and metastasis, the dielectric approach presented here provides a rapid and simple tool for screening the biophysical cell properties in studies on targeting chemo- or radiotherapeutically the

  12. Distribution of Prestin on Outer Hair Cell Basolateral Surface

    Institute of Scientific and Technical Information of China (English)

    YU Ning; ZHAI Suo-qiang; YANG Shi-ming; HAN Dong-yi; ZHAO Hong-bo

    2008-01-01

    Prestin has been identified as a motor protein responsible for outer hair cell (OHC) electromotility and is expressed on the OHC surface. Previous studies revealed that OHC eleetromotility and its associated nonlinear capacitance were mainly located at the OHC lateral wall and absent at the apical cutieular plate and the basal nucleus region. Immunofluorescent staining for prestin also failed to demonstrate prestin expression at the OHC basal ends in whole-mount preparation of the organ of Corti. However, there lacks a definitive demonstration of the pattern of prestin distribution. The OHC lateral wall has a trilaminate organization and is composed of the plasma membrane, cortical lattice, and subsurface cisternae. In this study, the location of prestin proteins in dissociated OHCs was examined using immunofluorescent staining and confocal microscopy. We found that prestin was uniformly expressed on the basolateral surface, including the basal pole. No staining was seen on the cuticular plate and stereocilia. When co-stained with a membrane marker di-8-ANEPPS, prestin-labeling was found to be in the outer layer of the OHC lateral wall. After separating the plasma membrane from the underlying subsurface eisternae using a hypotonic extracellular solution, prestin-labeling was found to be in the plasma membrane, not the subsurface cisternae. The data show that prestin is expressed in the plasma membrane on the entire OHC basolateral surface.

  13. Surface modification on a glass surface with a combination technique of sol–gel and air brushing processes

    KAUST Repository

    Tsai, Meng-Yu

    2011-08-01

    This study fabricated the large area and optically transparent superhydrophobic silica based films on glass surface with optimized hardness. A silane coupling agent, tetraethoxysilane (TEOS), effectively bonds silica particles onto the glass substrate. Desired surface roughness was obtained by adjusting nano silica particles concentration of the precursors prepared by the sol-gel process. Silica suspension was coated onto the glass substrate by the air brushing methods. This method can deposit a uniform, transparent coating on the glass substrate efficiently. Diluting the precursor by adding ethanol or a mixture of D.I. water and ethanol further improved the transmittance and superhydrophobicity efficiency. The results showed that as the silica particle concentration and the thickness of the coating were increased, the surface roughness was enhanced. Rougher surface displayed a higher superhydrophobicity and lower transmittance. Therefore, the concentration of silica particle, volume of coatings, and the ratio of ethanol and D.I. water are of great importance to deposit a transparent, superhydrophobic coating on glass. © 2011 Elsevier B.V. All rights reserved.

  14. Surface Analyses and Immune Reactivities of Major Cell Wall-Associated Proteins of Group A Streptococcus

    OpenAIRE

    Cole, Jason N; Ramirez, Ruben D.; Currie, Bart J.; Cordwell, Stuart J.; Djordjevic, Steven P.; Mark J Walker

    2005-01-01

    A proteomic analysis was undertaken to identify cell wall-associated proteins of Streptococcus pyogenes. Seventy-four distinct cell wall-associated proteins were identified, 66 of which were novel. Thirty-three proteins were immunoreactive with pooled S. pyogenes-reactive human antisera. Biotinylation of the GAS cell surface identified 23 cell wall-associated proteins that are surface exposed.

  15. Combination of genistein with ionizing radiation on andro-gen-independent prostate cancer cells

    Institute of Scientific and Technical Information of China (English)

    Sen-XiangYan; YasuoEjima; RyoheiSasaki; Shu-SenZheng; YusukeDemizu; ToshinoriSoejima; KazuroSugimura

    2004-01-01

    Aim: To study the effect of the combined use of genistein and ionizing radiation (IR) on prostate DUI45 cancer cells. Methods: DU145, an androgen-independent human prostate cancer cell line, was used in the experiment.Clonogenic assay was used to compare the survival of DU145 cells after treatments with genistein alone and in combination with graded IR. Apoptosis was assayed by DNA ladder and TUNEL stain. Cell cycle alterations were observed by flow cytometry and related protein expressions by immunoblotting. Results: Clonogenic assay demonstrated that genistein, even at low to medium concentrations, enhanced the radiosensitivity of DU 145 cells. Twenty four hours after treatment with IR and/or genistein, apoptosis was mainly seen with genistein at high concentrations and was minimally related to IR. At 72 h, apoptosis also occurred in treatment with lower concentration of genistein,especially when combined with IR. While both IR and genistein led to G2/M cell cycle arrest, combination of them further increased the DUI45 cells at G2/M phase. This G2/M arrest was largely maintained at 72 h, accompanied by increasing apoptosis and hyperdiploid cell population. Cell-cycle related protein analysis disclosed biphasic changes in cyclin B 1 and less dramatically cdc-2, but stably elevated p21cip1 levels with increasing genistein concentrations.Conclusion: Genistein enhanced the radiosensitivity of DUI45 prostate cancer cells. The mechanisms might be involved in the increased apoptosis, prolonged cell cycle arrest and impaired damage repair. (Asian J Androl 2004 Dec; 6: 285-290)

  16. Combination of Potassium Pentagamavunon-0 and Doxorubicin Induces Apoptosis and Cell Cycle Arrest and Inhibits Metastasis in Breast Cancer Cells.

    Science.gov (United States)

    Putri, Herwandhani; Jenie, Riris Istighfari; Handayani, Sri; Kastian, Ria Fajarwati; Meiyanto, Edy

    2016-01-01

    A salt compound of a curcumin analogue, potassium pentagamavunon-0 (K PGV-0) has been synthesized to improve solubility of pentagamavunon-0 which has been proven to have anti-proliferative effects on several cancer cells. The purpose of this study was to investigate cytotoxic activity and metastasis inhibition by K PGV- 0 alone and in combination with achemotherapeutic agent, doxorubicin (dox), in breast cancer cells. Based on MTT assay analysis, K PGV-0 showed cytotoxic activity in T47D and 4T1 cell lines with IC50 values of 94.9 μM and 49.0±0.2 μM, respectively. In general, K PGV-0+dox demonstrated synergistic effects and decreased cell viability up to 84.7% in T47D cells and 62.6% in 4T1 cells. Cell cycle modulation and apoptosis induction were examined by flow cytometry. K PGV-0 and K PGV-0+dox caused cell accumulation in G2/M phase and apoptosis induction. Regarding cancer metastasis, while K PGV-0 alone did not show any inhibition of 4T1 cell migration, K PGV-0+dox exerted inhibition. K PGV-0 and its combination with dox inhibited the activity of MMP-9 which has a pivotal role in extracellular matrix degradation. These results show that a combination of K PGV-0 and doxorubicin inhibits cancer cell growth through cell cycling, apoptosis induction, and inhibition of cell migration and MMP-9 activity. Therefore, K PGV-0 may have potential for development as a co-chemotherapeutic agent. PMID:27268651

  17. Altered expression of epithelial cell surface glycoconjugates and intermediate filaments at the margins of mucosal wounds

    DEFF Research Database (Denmark)

    Dabelsteen, Erik; Grøn, B; Mandel, U;

    1998-01-01

    Alterations in cell to cell adhesion are necessary to enable the type of cell movements that are associated with epithelial wound healing and malignant invasion. Several studies of transformed cells have related epithelial cell movement to changes in the cell surface expression of the carbohydrat...

  18. Wolbachia surface protein induces innate immune responses in mosquito cells

    Directory of Open Access Journals (Sweden)

    Pinto Sofia B

    2012-01-01

    Full Text Available Abstract Background Wolbachia endosymbiotic bacteria are capable of inducing chronic upregulation of insect immune genes in some situations and this phenotype may influence the transmission of important insect-borne pathogens. However the molecules involved in these interactions have not been characterized. Results Here we show that recombinant Wolbachia Surface Protein (WSP stimulates increased transcription of immune genes in mosquito cells derived from the mosquito Anopheles gambiae, which is naturally uninfected with Wolbachia; at least two of the upregulated genes, TEP1 and APL1, are known to be important in Plasmodium killing in this species. When cells from Aedes albopictus, which is naturally Wolbachia-infected, were challenged with WSP lower levels of upregulation were observed than for the An. gambiae cells. Conclusions We have found that WSP is a strong immune elicitor in a naturally Wolbachia-uninfected mosquito species (Anopheles gambiae while a milder elicitor in a naturally-infected species (Aedes albopictus. Since the WSP of a mosquito non-native (nematode Wolbachia strain was used, these data suggest that there is a generalized tolerance to WSP in Ae. albopictus.

  19. Soluble and cell surface receptors for tumor necrosis factor

    DEFF Research Database (Denmark)

    Wallach, D; Engelmann, H; Nophar, Y;

    1991-01-01

    Tumor necrosis factor (TNF) initiates its multiple effects on cell function by binding at a high affinity to specific cell surface receptors. Two different molecular species of these receptors, which are expressed differentially in different cells, have been identified. The cDNAs of both receptor...... have recently been cloned. Antibodies to one of these receptor species (the p55, type I receptor) can trigger a variety of TNF like effects by cross-linking of the receptor molecules. Thus, it is not TNF itself but its receptors that provide the signal for the response to this cytokine....... The intracellular domains of the two receptors differ in structure, suggesting that they mediate different activities. Their extracellular domains, however, are structurally related. Both contain cysteine-rich repeats which are homologous to repeated structures found in the extracellular domains of the nerve growth...... factor receptor and the CDw40 protein. Truncated soluble forms of the two receptors, corresponding to these cysteine-rich repeated structures, have been detected in human urine and were later found to be present also in the serum. The serum levels of those soluble TNF receptors increase dramatically...

  20. Regulation of ADAM12 cell-surface expression by protein kinase C epsilon

    DEFF Research Database (Denmark)

    Sundberg, Christina; Thodeti, Charles Kumar; Kveiborg, Marie;

    2004-01-01

    constitutively active protein. However, little is known about the regulation of ADAM12 cell-surface translocation. Here, we used human RD rhabdomyosarcoma cells, which express ADAM12 at the cell surface, in a temporal pattern. We report that protein kinase C (PKC) epsilon induces ADAM12 translocation to the cell......The ADAM (a disintegrin and metalloprotease) family consists of multidomain cell-surface proteins that have a major impact on cell behavior. These transmembrane-anchored proteins are synthesized as proforms that have (from the N terminus): a prodomain; a metalloprotease-, disintegrin......-immunoprecipitated from membrane-enriched fractions of PMA-treated cells, 3) RD cells transfected with EGFP-tagged, myristoylated PKCepsilon expressed more ADAM12 at the cell surface than did non-transfected cells, and 4) RD cells transfected with a kinase-inactive PKCepsilon mutant did not exhibit ADAM12 cell...

  1. VUV treatment combined with mechanical strain of stretchable polymer foils resulting in cell alignment

    Energy Technology Data Exchange (ETDEWEB)

    Barb, R.-A. [Institute of Applied Physics, Johannes Kepler University Linz (Austria); Magnus, B. [Innovacell Biotechnologie AG, Innsbruck (Austria); Innerbichler, S. [Innerbichler GmbH, Breitenbach am Inn (Austria); Greunz, T. [CDL-MS-MACH, Johannes Kepler University Linz (Austria); Wiesbauer, M. [Institute of Applied Physics, Johannes Kepler University Linz (Austria); Marksteiner, R. [Innovacell Biotechnologie AG, Innsbruck (Austria); Stifter, D. [CDL-MS-MACH, Johannes Kepler University Linz (Austria); Heitz, J., E-mail: johannes.heitz@jku.at [Institute of Applied Physics, Johannes Kepler University Linz (Austria)

    2015-01-15

    Highlights: • Elastic polyurethane (PU) foils were exposed to the vacuum-UV in reactive atmosphere. • The photomodification resulted in improved cytocompatibilty. • Parallel microgrooves formed on the irradiated PU surfaces after strong elongation. • Cells seeded onto microgrooves aligned their shapes in the direction of the grooves. • Elongation occurred also for cells on PU subjected to cyclic mechanical stretching. - Abstract: Cell-alignment along a defined direction can have a direct effect on the cell functionality and differentiation. Oriented micro- or nanotopographic structures on cell culture substrates can induce cell-alignment. Surface chemistry, wettability, and stiffness of the substrate are also important material features as they strongly influence the cell–substrate interactions. For improved bio-compatibility, highly elastic polyurethane (PU) foils were exposed to the vacuum-UV (VUV) light of a Xe{sub 2}{sup *} excimer lamp at 172 nm in a nitrogen containing atmosphere (N{sub 2} or NH{sub 3}). The irradiation resulted in a change in the chemical surface composition. Additionally, the formation of regular parallel microgrooves was observed on the irradiated surfaces after strong uni-axial deformation (i.e., more than about 50% strain) of the photo-modified PU foils. Cell seeding experiments demonstrated that the VUV modified polymer foils strongly enhance cell adhesion and proliferation. Cells seeded onto microgrooves aligned their shapes and elongated in the direction of the grooves. A similar effect was observed for cells seeded on photo-modified PU foils subjected to cyclic mechanical stretching at lower strain levels (i.e., typically 10% strain) without groove-formation. The cells had also here an elongated shape, however they not always align in a defined direction relative to the stretching.

  2. Effective combination treatment of lung cancer cells by single vehicular delivery of siRNA and different anticancer drugs

    Science.gov (United States)

    Li, Jinming; Wang, Yuanyuan; Xue, Shanshan; Sun, Jinghua; Zhang, Wei; Hu, Ping; Ji, Liangnian; Mao, Zongwan

    2016-01-01

    In recent years, lung cancer has become one of the fastest growing cancers in the world. Thus, the development of efficient combination therapy to treat lung cancer has attracted significant attention in the cancer therapy field. In this article, we developed a single vehicle drug delivery system, based on quantum dot (QD) nanoparticles, to deliver small interfering RNA (siRNA; target Bcl-2) and different anticancer drugs (carboplatin, paclitaxel, and doxorubicin) simultaneously for treating A549 lung cancer cells efficiently by combination therapy. The QD nanoparticles were conjugated with l-arginine (l-Arg) and different kinds of hydroxypropyl-cyclodextrins (HP-α-CDs, HP-β-CDs, and HP-γ-CDs) on the surface to form the delivery nanocarriers (QD nanocarriers). They were able to not only bind and transport the siRNA through electrostatic interactions with l-Arg residues but also accommodate various disparate anticancer drugs using different HP-CD modifications. Compared with free drug treatments, the use of QD nanocarriers to deliver Bcl-2 siRNA and different anticancer drugs simultaneously exerted a threefold to fourfold increase in cytotoxicity in A549 cells, which greatly improved the treatment efficacy through combined action. Furthermore, the QD nanocarriers could be used as a probe for real-time imaging of the drug delivery and release because of their strong fluorescence properties. These findings indicate that multifunctional QD nanocarriers hold great promise as a powerful tool for combination therapy for lung cancer.

  3. Effects of the combination of decitabine and homoharringtonine in SKM-1 and Kg-1a cells.

    Science.gov (United States)

    Geng, Suxia; Yao, Han; Weng, Jianyu; Tong, Jiaqi; Huang, Xin; Wu, Ping; Deng, Chengxin; Li, Minming; Lu, Zesheng; Du, Xin

    2016-05-01

    The methylation inhibitor decitabine (DAC) has great therapeutic value for myelodysplastic syndrome (MDS) and acute myeloid leukemia (AML). However, DAC monotherapy is associated with relatively low rates of overall response and complete remission. Previous studies have shown promising results for combination treatment regimens including DAC. Homoharringtonine (HHT), an alkaloid from Chinese natural plants and Cephalotaxus, has demonstrated potential for leukemia treatment. Our studies have suggested that the combination of DAC and HHT has synergistic effects for inhibiting the viability of SKM-1 and Kg-1a cells. This combination leads to enhanced inhibition of colony formation and apoptosis induction compared with DAC alone in SKM-1 but not Kg-1a cells. Only high-dose DAC and HHT significantly up-regulate caspase-3 and caspase-9 and inhibit BCL-XL in the SKM-1 cell line. The combined effects of DAC plus HHT on apoptosis may not only depend on regulation of the apoptosis-related genes we examined but others as well. HHT had no demethylation effects, and HHT in combination with DAC had no enhanced effects on hypomethylation and DNMT1, DNMT3A and DNMT3B mRNA expression in SKM-1 cells. Overall, these results suggest that DAC used in combination with HHT may have clinical potential for MDS treatment.

  4. Cell surface glycan alterations in epithelial mesenchymal transition process of Huh7 hepatocellular carcinoma cell.

    Directory of Open Access Journals (Sweden)

    Shan Li

    Full Text Available BACKGROUND AND OBJECTIVE: Due to recurrence and metastasis, the mortality of Hepatocellular carcinoma (HCC is high. It is well known that the epithelial mesenchymal transition (EMT and glycan of cell surface glycoproteins play pivotal roles in tumor metastasis. The goal of this study was to identify HCC metastasis related differential glycan pattern and their enzymatic basis using a HGF induced EMT model. METHODOLOGY: HGF was used to induce HCC EMT model. Lectin microarray was used to detect the expression of cell surface glycan and the difference was validated by lectin blot and fluorescence cell lectin-immunochemistry. The mRNA expression levels of glycotransferases were determined by qRT-PCR. RESULTS: After HGF treatment, the Huh7 cell lost epithelial characteristics and obtained mesenchymal markers. These changes demonstrated that HGF could induce a typical cell model of EMT. Lectin microarray analysis identified a decreased affinity in seven lectins ACL, BPL, JAC, MPL, PHA-E, SNA, and SBA to the glycan of cell surface glycoproteins. This implied that glycan containing T/Tn-antigen, NA2 and bisecting GlcNAc, Siaα2-6Gal/GalNAc, terminal α or βGalNAc structures were reduced. The binding ability of thirteen lectins, AAL, LCA, LTL, ConA, NML, NPL, DBA, HAL, PTL II, WFL, ECL, GSL II and PHA-L to glycan were elevated, and a definite indication that glycan containing terminal αFuc and ± Sia-Le, core fucose, α-man, gal-β(α GalNAc, β1,6 GlcNAc branching and tetraantennary complex oligosaccharides structures were increased. These results were further validated by lectin blot and fluorescence cell lectin-immunochemistry. Furthermore, the mRNA expression level of Mgat3 decreased while that of Mgat5, FucT8 and β3GalT5 increased. Therefore, cell surface glycan alterations in the EMT process may coincide with the expression of glycosyltransferase. CONCLUSIONS: The findings of this study systematically clarify the alterations of cell surface

  5. Near-infrared light triggered photodynamic therapy in combination with gene therapy using upconversion nanoparticles for effective cancer cell killing

    Science.gov (United States)

    Wang, Xin; Liu, Kai; Yang, Guangbao; Cheng, Liang; He, Lu; Liu, Yumeng; Li, Yonggang; Guo, Liang; Liu, Zhuang

    2014-07-01

    Upconversion nanoparticles (UCNPs) have drawn much attention in cancer imaging and therapy in recent years. Herein, we for the first time report the use of UCNPs with carefully engineered surface chemistry for combined photodynamic therapy (PDT) and gene therapy of cancer. In our system, positively charged NaGdF4:Yb,Er UCNPs with multilayered polymer coatings are synthesized via a layer by layer strategy, and then loaded simultaneously with Chlorin e6 (Ce6), a photosensitizing molecule, and small interfering RNA (siRNA), which targets the Plk1 oncogene. On the one hand, under excitation by a near-infrared (NIR) light at 980 nm, which shows greatly improved tissue penetration compared with visible light, cytotoxic singlet oxygen can be generated via resonance energy transfer from UCNPs to photosensitizer Ce6, while the residual upconversion luminescence is utilized for imaging. On the other hand, the silencing of Plk1 induced by siRNA delivered with UCNPs could induce significant cancer cell apoptosis. As the result of such combined photodynamic and gene therapy, a remarkably enhanced cancer cell killing effect is realized. Our work thus highlights the promise of UCNPs for imaging guided combination therapy of cancer.Upconversion nanoparticles (UCNPs) have drawn much attention in cancer imaging and therapy in recent years. Herein, we for the first time report the use of UCNPs with carefully engineered surface chemistry for combined photodynamic therapy (PDT) and gene therapy of cancer. In our system, positively charged NaGdF4:Yb,Er UCNPs with multilayered polymer coatings are synthesized via a layer by layer strategy, and then loaded simultaneously with Chlorin e6 (Ce6), a photosensitizing molecule, and small interfering RNA (siRNA), which targets the Plk1 oncogene. On the one hand, under excitation by a near-infrared (NIR) light at 980 nm, which shows greatly improved tissue penetration compared with visible light, cytotoxic singlet oxygen can be generated via

  6. Cell surface N-glycans influence the level of functional E-cadherin at the cell–cell border

    OpenAIRE

    M Kristen Hall; Douglas A Weidner; Sahil Dayal; Ruth A. Schwalbe

    2014-01-01

    E-cadherin is crucial for adhesion of cells to each other and thereby development and maintenance of tissue. While it is has been established that N-glycans inside the cell impact the level of E-cadherin at the cell surface of epithelial-derived cells, it is unclear whether N-glycans outside the cell control the clustering of E-cadherin at the cell–cell border. Here, we demonstrate reduction of N-glycans at the cell surface weakened the recruitment and retention of E-cadherin at the cell–cell...

  7. Combining surface plasmonic and light extraction enhancement on InGaN quantum-well light-emitters

    DEFF Research Database (Denmark)

    Fadil, Ahmed; Ou, Yiyu; Iida, Daisuke;

    2016-01-01

    and internal quantum efficiency enhancement for InGaN/GaN quantum-well light-emitters. By fabricating dielectric nano-rod pattern on the GaN surface, an optical coating that improves the light extraction is obtained, and furthermore has a low refractive index which blue-shifts the plasmonic resonance of Ag NPs...... towards the emission wavelength. We investigate emission components from both the GaN and sapphire surface of the semiconductor crystal and show that Ag NPs on dielectric nano-pattern compared to a planar surface, result in a stronger enhancement......Surface plasmon coupling with light-emitters and surface nano-patterning have widely been used separately to improve low efficiency InGaN light-emitting diodes. We demonstrate a method where dielectric nano-patterning and Ag nanoparticles (NPs) are combined to provide both light extraction...

  8. Combining phenotypic and proteomic approaches to identify membrane targets in a ‘triple negative’ breast cancer cell type

    Directory of Open Access Journals (Sweden)

    Rust Steven

    2013-02-01

    Full Text Available Abstract Background The continued discovery of therapeutic antibodies, which address unmet medical needs, requires the continued discovery of tractable antibody targets. Multiple protein-level target discovery approaches are available and these can be used in combination to extensively survey relevant cell membranomes. In this study, the MDA-MB-231 cell line was selected for membranome survey as it is a ‘triple negative’ breast cancer cell line, which represents a cancer subtype that is aggressive and has few treatment options. Methods The MDA-MB-231 breast carcinoma cell line was used to explore three membranome target discovery approaches, which were used in parallel to cross-validate the significance of identified antigens. A proteomic approach, which used membrane protein enrichment followed by protein identification by mass spectrometry, was used alongside two phenotypic antibody screening approaches. The first phenotypic screening approach was based on hybridoma technology and the second was based on phage display technology. Antibodies isolated by the phenotypic approaches were tested for cell specificity as well as internalisation and the targets identified were compared to each other as well as those identified by the proteomic approach. An anti-CD73 antibody derived from the phage display-based phenotypic approach was tested for binding to other ‘triple negative’ breast cancer cell lines and tested for tumour growth inhibitory activity in a MDA-MB-231 xenograft model. Results All of the approaches identified multiple cell surface markers, including integrins, CD44, EGFR, CD71, galectin-3, CD73 and BCAM, some of which had been previously confirmed as being tractable to antibody therapy. In total, 40 cell surface markers were identified for further study. In addition to cell surface marker identification, the phenotypic antibody screening approaches provided reagent antibodies for target validation studies. This is illustrated

  9. Studies on cell adhesion and recognition. II. The kinetics of cell adhesion and cell spreading on surfaces coated with carbohydrate- reactive proteins (glycosidases and lectins) and fibronectin

    OpenAIRE

    1981-01-01

    The kinetics of cell attachment and cell spreading on the coated surfaces of two classes of carbohydrate-reactive proteins, enzymes and lectins, have been compared with those on fibronectin-coated surfaces with the following results: (a) A remarkable similarity between the kinetics of cell attachment to fibronectin-coated and glycosidase- coated surfaces was found. In contrast, cell attachment kinetics induced by lectin- and galactose oxidase-coated surfaces, in general, were strikingly diffe...

  10. Distinct genetic alterations occur in ovarian tumor cells selected for combined resistance to carboplatin and docetaxel

    Directory of Open Access Journals (Sweden)

    Armstrong Stephen R

    2012-11-01

    Full Text Available Abstract Background Current protocols for the treatment of ovarian cancer include combination chemotherapy with a platinating agent and a taxane. However, many patients experience relapse of their cancer and the development of drug resistance is not uncommon, making successful second line therapy difficult to achieve. The objective of this study was to develop and characterize a cell line resistant to both carboplatin and docetaxel (dual drug resistant ovarian cell line and to compare this cell line to cells resistant to either carboplatin or docetaxel. Methods The A2780 epithelial endometrioid ovarian cancer cell line was used to select for isogenic carboplatin, docetaxel and dual drug resistant cell lines. A selection method of gradually increasing drug doses was implemented to avoid clonal selection. Resistance was confirmed using a clonogenic assay. Changes in gene expression associated with the development of drug resistance were determined by microarray analysis. Changes in the expression of selected genes were validated by Quantitative Real-Time Polymerase Chain Reaction (QPCR and immunoblotting. Results Three isogenic cell lines were developed and resistance to each drug or the combination of drugs was confirmed. Development of resistance was accompanied by a reduced growth rate. The microarray and QPCR analyses showed that unique changes in gene expression occurred in the dual drug resistant cell line and that genes known to be involved in resistance could be identified in all cell lines. Conclusions Ovarian tumor cells can acquire resistance to both carboplatin and docetaxel when selected in the presence of both agents. Distinct changes in gene expression occur in the dual resistant cell line indicating that dual resistance is not a simple combination of the changes observed in cell lines exhibiting single agent resistance.

  11. Interaction of progenitor bone cells with different surface modifications of titanium implant

    Energy Technology Data Exchange (ETDEWEB)

    Chen, Wen-Cheng, E-mail: wencchen@fcu.edu.tw [Advanced Medical Devices and Composites Laboratory, Department of Fiber and Composite Materials, College of Engineering, Feng Chia University, Taichung 40724, Taiwan (China); Chen, Ya-Shun [Advanced Medical Devices and Composites Laboratory, Department of Fiber and Composite Materials, College of Engineering, Feng Chia University, Taichung 40724, Taiwan (China); Ko, Chia-Ling [Advanced Medical Devices and Composites Laboratory, Department of Fiber and Composite Materials, College of Engineering, Feng Chia University, Taichung 40724, Taiwan (China); Dental Medical Devices and Materials Research Center, College of Dental Medicine, Kaohsiung Medical University, Kaohsiung 807, Taiwan (China); Lin, Yi; Kuo, Tzu-Huang; Kuo, Hsien-Nan [Medical Device Development Division, Metal Industries Research and Development Centre, Kaohsiung 82151, Taiwan (China)

    2014-04-01

    Changes in the physical and chemical properties of Ti surfaces can be attributed to cell performance, which improves surface biocompatibility. The cell proliferation, mineralization ability, and gene expression of progenitor bone cells (D1 cell) were compared on five different Ti surfaces, namely, mechanical grinding (M), electrochemical modification through potentiostatic anodization (ECH), sandblasting and acid etching (SLA), sandblasting, hydrogen peroxide treatment, and heating (SAOH), and sandblasting, alkali heating, and etching (SMART). SAOH treatment produced the most hydrophilic surface, whereas SLA produced the most hydrophobic surface. Cell activity indicated that SLA and SMART produced significantly rougher surfaces and promoted D1 cell attachment within 1 day of culturing, whereas SAOH treatment produced moderate roughness (Ra = 1.26 μm) and accelerated the D1 cell proliferation up to 7 days after culturing. The ECH surface significantly promoted alkaline phosphatase (ALP) expression and osteocalcin (OCN) secretion in the D1 cells compared with the other surface groups. The ECH and SMART-treated Ti surfaces resulted in maximum ALP and OCN expressions during the D1 cell culture. SLA, SAOH, and SMART substrate surfaces were rougher and exhibited better cell metabolic responses during the early stage of cell attachment, proliferation, and morphologic expressions within 1 day of D1 cell culture. The D1 cells cultured on the ECH and SMART substrates exhibited higher differentiation, and higher ALP and OCN expressions after 10 days of culture. Thus, the ECH and SMART treatments promote better ability of cell mineralization in vitro, which demonstrate their great potential for clinical use. - Highlights: • Progenitor bone cells onto Ti with different modifications are characterized. • Surface roughness and hydrophilicity encourage early stage cell attachment. • Composition and surface treatments are more vital in bone cell mineralization.

  12. Interaction of progenitor bone cells with different surface modifications of titanium implant

    International Nuclear Information System (INIS)

    Changes in the physical and chemical properties of Ti surfaces can be attributed to cell performance, which improves surface biocompatibility. The cell proliferation, mineralization ability, and gene expression of progenitor bone cells (D1 cell) were compared on five different Ti surfaces, namely, mechanical grinding (M), electrochemical modification through potentiostatic anodization (ECH), sandblasting and acid etching (SLA), sandblasting, hydrogen peroxide treatment, and heating (SAOH), and sandblasting, alkali heating, and etching (SMART). SAOH treatment produced the most hydrophilic surface, whereas SLA produced the most hydrophobic surface. Cell activity indicated that SLA and SMART produced significantly rougher surfaces and promoted D1 cell attachment within 1 day of culturing, whereas SAOH treatment produced moderate roughness (Ra = 1.26 μm) and accelerated the D1 cell proliferation up to 7 days after culturing. The ECH surface significantly promoted alkaline phosphatase (ALP) expression and osteocalcin (OCN) secretion in the D1 cells compared with the other surface groups. The ECH and SMART-treated Ti surfaces resulted in maximum ALP and OCN expressions during the D1 cell culture. SLA, SAOH, and SMART substrate surfaces were rougher and exhibited better cell metabolic responses during the early stage of cell attachment, proliferation, and morphologic expressions within 1 day of D1 cell culture. The D1 cells cultured on the ECH and SMART substrates exhibited higher differentiation, and higher ALP and OCN expressions after 10 days of culture. Thus, the ECH and SMART treatments promote better ability of cell mineralization in vitro, which demonstrate their great potential for clinical use. - Highlights: • Progenitor bone cells onto Ti with different modifications are characterized. • Surface roughness and hydrophilicity encourage early stage cell attachment. • Composition and surface treatments are more vital in bone cell mineralization.

  13. Apicidin and Docetaxel Combination Treatment Drives CTCFL Expression and HMGB1 Release Acting as Potential Antitumor Immune Response Inducers in Metastatic Breast Cancer Cells

    Directory of Open Access Journals (Sweden)

    Maria Buoncervello

    2012-09-01

    Full Text Available Currently approved combination regimens available for the treatment of metastatic tumors, such as breast cancer, have been shown to increase response rates, often at the cost of a substantial increase in toxicity. An ideal combination strategy may consist of agents with different mechanisms of action leading to complementary antitumor activities and safety profiles. In the present study, we investigated the effects of the epigenetic modulator apicidin in combination with the cytotoxic agent docetaxel in tumor breast cell lines characterized by different grades of invasiveness. We report that combined treatment of apicidin and docetaxel, at low toxicity doses, stimulates in metastatic breast cancer cells the expression of CTCF-like protein and other cancer antigens, thus potentially favoring an antitumor immune response. In addition, apicidin and docetaxel co-treatment specifically stimulates apoptosis, characterized by an increased Bax/Bcl-2 ratio and caspase-8 activation. Importantly, following combined exposure to these agents, metastatic cells were also found to induce signals of immunogenic apoptosis such as cell surface expression of calreticulin and release of considerable amounts of high-mobility group box 1 protein, thus potentially promoting the translation of induced cell death into antitumor immune response. Altogether, our results indicate that the combined use of apicidin and docetaxel, at a low toxicity profile, may represent a potential innovative strategy able to activate complementary antitumor pathways in metastatic breast cancer cells, associated with a potential control of metastatic growth and possible induction of antitumor immunity.

  14. Controlled surface modification of tissue culture polystyrene for selective cell binding using resilin-inspired polypeptides

    International Nuclear Information System (INIS)

    Modified tissue culture polystyrene (TCP) surfaces have been fabricated by attachment of recombinant polypeptides based on Drosophila melanogaster resilin and the Anopheles gambiae resilin-like protein. The D. melanogaster polypeptide (Rec-1) was from the first exon of resilin and consisted of 17 very similar repeats of a 15 residue sequence. The A. gambiae polypeptide consisted of 16 repeats of an 11 residue consensus sequence (An16). Polypeptides were attached to the TCP surface through tyrosine-based photo-crosslinking using blue light in combination with (RuII(bpy)3)Cl2 and sodium persulfate. TCP that has been manufactured by mild oxidation has surface phenolic groups that are believed to participate in this crosslinking process. X-ray photoelectron spectroscopy and contact angle analyses were used to demonstrate polypeptide binding. At higher coating concentrations of Rec-1 and An16, the surface was passivated and fibroblasts no longer attached and spread. At coating concentrations of 1 mg ml−1 for Rec-1 and 0.1 mg ml−1 for An16, where the surface was fully passivated against fibroblast attachment, addition of a cell attachment peptide, cyclo(Arg-Gly-Asp-D-Tyr-Lys) during coating and photo-crosslinking at >0.1 mg ml−1, led to the restoration of fibroblast binding that was dependent on the integrin αV chain. (paper)

  15. Synergistic inhibition of cancer cell proliferation with a combination of δ-tocotrienol and ferulic acid

    Energy Technology Data Exchange (ETDEWEB)

    Eitsuka, Takahiro, E-mail: eitsuka@nupals.ac.jp [Faculty of Applied Life Sciences, Niigata University of Pharmacy and Applied Life Sciences, Niigata 956-8603 (Japan); Tatewaki, Naoto; Nishida, Hiroshi; Kurata, Tadao [Faculty of Applied Life Sciences, Niigata University of Pharmacy and Applied Life Sciences, Niigata 956-8603 (Japan); Nakagawa, Kiyotaka; Miyazawa, Teruo [Food and Biodynamic Chemistry Laboratory, Graduate School of Agricultural Science, Tohoku University, Sendai 981-8555 (Japan)

    2014-10-24

    Highlights: • δ-Tocotrienol (δ-T3) and ferulic acid (FA) synergistically inhibit cancer cell growth. • The combination of δ-T3 and FA induces G1 arrest by up-regulating p21. • The synergy is attributed to an increase in the cellular concentration of δ-T3 by FA. - Abstract: Rice bran consists of many functional compounds and thus much attention has been focused on the health benefits of its components. Here, we investigated the synergistic inhibitory effects of its components, particularly δ-tocotrienol (δ-T3) and ferulic acid (FA), against the proliferation of an array of cancer cells, including DU-145 (prostate cancer), MCF-7 (breast cancer), and PANC-1 (pancreatic cancer) cells. The combination of δ-T3 and FA markedly reduced cell proliferation relative to δ-T3 alone, and FA had no effect when used alone. Although δ-T3 induced G1 arrest by up-regulating p21 in PANC-1 cells, more cells accumulated in G1 phase with the combination of δ-T3 and FA. This synergistic effect was attributed to an increase in the cellular concentration of δ-T3 by FA. Our results suggest that the combination of δ-T3 and FA may present a new strategy for cancer prevention and therapy.

  16. T CELL REPERTOIRE COMPLEXITY IN SEVER COMBINED IMMUNODEFICIENCY PATIENTS AFTER BONE MARROW TRANSPLANTATION

    Institute of Scientific and Technical Information of China (English)

    曹水; 李晓静

    2003-01-01

    Objective. To study thymus-dependent T cell development and T cell repertoire in human sever combined immunodeficiency (SCID) patients after HLA-identical or haploidentical T cell-depleted allogeneic bone marrow transplantation (BMT). Methods. Blood samples were obtained from 15 SCID patients before transplantation and at varying intervals thereafter. Quantitative competitive PCR assay and immunoscope analysis of the T cell receptor (TCR) Vβ repertoire were performed. Results. Before and within the first 100 days after transplantation, patients' peripheral blood mononuclear cell (PBMC) presented an oligoclonal or polyclonal skewed T cell repertoire, low T cell receptor excision circles (TRECs) values and predominance of CD45RO+ T cell. In contrast, the presence of high numbers of CD45RA+ T cells in bone marrow(BM) circulation reconstituted SCID patients (>100 days post-transplantation) correlated with active T cell production by the thymus as revealed by high TREC values, and a polyclonal T cell repertoire demonstrated by a Gaussian distribution of Vβ-specific peaks. Conclusions. Within one year after BMT, a normal T cell repertoire develops in SCID patients as a result of thymic output. The T cell receptor diversity is highly and positively correlated in these patients with TREC levels.

  17. Microfluidic-based single cell trapping using a combination of stagnation point flow and physical barrier

    Science.gov (United States)

    Yu, Miao; Chen, Zongzheng; Xiang, Cheng; Liu, Bo; Xie, Handi; Qin, Kairong

    2016-06-01

    Single cell trapping in vitro by microfluidic device is an emerging approach for the study of the relationship between single cells and their dynamic biochemical microenvironments. In this paper, a hydrodynamic-based microfluidic device for single cell trapping is designed using a combination of stagnation point flow and physical barrier. The microfluidic device overcomes the weakness of the traditional ones, which have been only based upon either stagnation point flows or physical barriers, and can conveniently load dynamic biochemical signals to the trapped cell. In addition, it can connect with a programmable syringe pump and a microscope to constitute an integrated experimental system. It is experimentally verified that the microfluidic system can trap single cells in vitro even under flow disturbance and conveniently load biochemical signals to the trapped cell. The designed micro-device would provide a simple yet effective experimental platform for further study of the interactions between single cells and their microenvironments.

  18. Synergistic effect of combining paeonol and cisplatin on apoptotic induction of human hepatoma cell lines

    Institute of Scientific and Technical Information of China (English)

    Shu-ping XU; Guo-ping SUN; Yu-xian SHEN; Wan-ten PENG; Hua WANG; Wei WE

    2007-01-01

    Aim: To investigate whether paeonol (Pae) has synergistic effects with cisplatin (CDDP) on the growth-inhibition and apoptosis-induction of human hepatoma cell lines HepG2 and SMMC-7721.Methods: The cytotoxic effect of drugs was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide assay. The coefficient of drug interaction was used to analyze the nature of drug interactions. Morphological changes were observed by acridine orange fluo-rescence staining. Cell cycle and the apoptosis rate were detected by flow cytometry. Bcl-2 and Bax expression were assayed by immunohistochemical staining.Results: Pae or CDDP had antiproliferative effect on the 2 cell lines in a dose-dependent manner, with different sensitivities to drugs. More interestingly, a synergistic inhibitory effect on the viability of the 2 cell lines was observed after treatment with a combination of Pae (15.63, 31.25, and 62.5 mg/L) with various concentrations of CDDP. Further study showed typical mor-phological changes of apoptosis if the cells were exposed to the two agents for 24 h. The apoptotic rate of the cells with combination treatment was signifi-candy higher than that of cells treated with Pae or CDDP alone. The expression of Bcl-2 decreased and that of Bax increased in the treated groups, especially in the combination group, with the ratio of Bcl-2/Bax decreasing correspondingly.Additionally, a combination of Pae with CDDP resulted in a stronger S phase arrest, compared with Pae or CDDP alone.Conclusion: Pae, in combination with CDDP, had a significantly synergistic growth-inhibitory and apoptosis-in-ducing effect on the 2 human hepatoma cell lines, which may be useful in hepatoma treatment.

  19. Simultaneous Retrieval of Aerosol and Surface Optical Properties from Combined Airborne- and Ground-Based Direct and Diffuse Radiometric Measurements

    Science.gov (United States)

    Gatebe, C. K.; Dubovik, O.; King, M. D.; Sinyuk, A.

    2010-01-01

    This paper presents a new method for simultaneously retrieving aerosol and surface reflectance properties from combined airborne and ground-based direct and diffuse radiometric measurements. The method is based on the standard Aerosol Robotic Network (AERONET) method for retrieving aerosol size distribution, complex index of refraction, and single scattering albedo, but modified to retrieve aerosol properties in two layers, below and above the aircraft, and parameters on surface optical properties from combined datasets (Cloud Absorption Radiometer (CAR) and AERONET data). A key advantage of this method is the inversion of all available spectral and angular data at the same time, while accounting for the influence of noise in the inversion procedure using statistical optimization. The wide spectral (0.34-2.30 m) and angular range (180 ) of the CAR instrument, combined with observations from an AERONET sunphotometer, provide sufficient measurement constraints for characterizing aerosol and surface properties with minimal assumptions. The robustness of the method was tested on observations made during four different field campaigns: (a) the Southern African Regional Science Initiative 2000 over Mongu, Zambia, (b) the Intercontinental Transport Experiment-Phase B over Mexico City, Mexico (c) Cloud and Land Surface Interaction Campaign over the Atmospheric Radiation Measurement (ARM) Central Facility, Oklahoma, USA, and (d) the Arctic Research of the Composition of the Troposphere from Aircraft and Satellites (ARCTAS) over Elson Lagoon in Barrow, Alaska, USA. The four areas are dominated by different surface characteristics and aerosol types, and therefore provide good test cases for the new inversion method.

  20. Cell adhesion behavior on the silicone rubber surface modified by using ion beam irradiation

    Energy Technology Data Exchange (ETDEWEB)

    Hwang, In Tae; Jung, Chan Hee; Nh, Young Chang; Choi, Jae Hak [Korea Atomic Energy Research Institute, Daejeon (Korea, Republic of); Kuk, In Seol [Hanyang University, Seoul (Korea, Republic of); An, Mi Young [Chungnam National University, Daejeon (Korea, Republic of)

    2009-12-15

    In this study we studied cell adhesion and proliferation on the surface of a silicone rubber modified by ion beam irradiation. The surface property of the irradiated silicone rubber was characterized by water contact angle and FT-IR analyses. It was observed that human (HEK293) fibroblast cells exhibit strong adhesion to the irradiated silicone surface. This enhanced adhesion of mammalian cells can be attributed to the increase in the hydrophilicity of the silicone surface by ion beam irradiation.

  1. A combination of an anti-SLAMF6 antibody and ibrutinib efficiently abrogates expansion of chronic lymphocytic leukemia cells

    Science.gov (United States)

    Yigit, Burcu; Halibozek, Peter J.; Chen, Shih-Shih; O'Keeffe, Michael S.; Arnason, Jon; Avigan, David; Gattei, Valter; Bhan, Atul; Cen, Osman; Longnecker, Richard; Chiorazzi, Nicholas; Wang, Ninghai; Engel, Pablo; Terhorst, Cox

    2016-01-01

    The signaling lymphocyte activation molecule family [SLAMF] of cell surface receptors partakes in both the development of several immunocyte lineages and innate and adaptive immune responses in humans and mice. For instance, the homophilic molecule SLAMF6 (CD352) is in part involved in natural killer T cell development, but also modulates T follicular helper cell and germinal B cell interactions. Here we report that upon transplantation of a well-defined aggressive murine B220+CD5+ Chronic Lymphocytic Leukemia (CLL) cell clone, TCL1-192, into SCID mice one injection of a monoclonal antibody directed against SLAMF6 (αSlamf6) abrogates tumor progression in the spleen, bone marrow and blood. Similarly, progression of a murine B cell lymphoma, LMP2A/λMyc, was also eliminated by αSlamf6. But, surprisingly, αSLAMF6 neither eliminated TCL1-192 nor LMP2A/λMyc cells, which resided in the peritoneal cavity or omentum. This appeared to be dependent upon the tumor environment, which affected the frequency of sub-populations of the TCL1-192 clone or the inability of peritoneal macrophages to induce Antibody Dependent Cellular Cytotoxicity (ADCC). However, co-administering αSlamf6 with the Bruton tyrosine kinase (Btk) inhibitor, ibrutinib, synergized to efficiently eliminate the tumor cells in the spleen, bone marrow, liver and the peritoneal cavity. Because an anti-human SLAMF6 mAb efficiently killed human CLL cells in vitro and in vivo, we propose that a combination of αSlamf6 with ibrutinib should be considered as a novel therapeutic approach for CLL and other B cell tumors. PMID:27029059

  2. A combination of an anti-SLAMF6 antibody and ibrutinib efficiently abrogates expansion of chronic lymphocytic leukemia cells.

    Science.gov (United States)

    Yigit, Burcu; Halibozek, Peter J; Chen, Shih-Shih; O'Keeffe, Michael S; Arnason, Jon; Avigan, David; Gattei, Valter; Bhan, Atul; Cen, Osman; Longnecker, Richard; Chiorazzi, Nicholas; Wang, Ninghai; Engel, Pablo; Terhorst, Cox

    2016-05-01

    The signaling lymphocyte activation molecule family [SLAMF] of cell surface receptors partakes in both the development of several immunocyte lineages and innate and adaptive immune responses in humans and mice. For instance, the homophilic molecule SLAMF6 (CD352) is in part involved in natural killer T cell development, but also modulates T follicular helper cell and germinal B cell interactions. Here we report that upon transplantation of a well-defined aggressive murine B220+CD5+ Chronic Lymphocytic Leukemia (CLL) cell clone, TCL1-192, into SCID mice one injection of a monoclonal antibody directed against SLAMF6 (αSlamf6) abrogates tumor progression in the spleen, bone marrow and blood. Similarly, progression of a murine B cell lymphoma, LMP2A/λMyc, was also eliminated by αSlamf6. But, surprisingly, αSLAMF6 neither eliminated TCL1-192 nor LMP2A/λMyc cells, which resided in the peritoneal cavity or omentum. This appeared to be dependent upon the tumor environment, which affected the frequency of sub-populations of the TCL1-192 clone or the inability of peritoneal macrophages to induce Antibody Dependent Cellular Cytotoxicity (ADCC). However, co-administering αSlamf6 with the Bruton tyrosine kinase (Btk) inhibitor, ibrutinib, synergized to efficiently eliminate the tumor cells in the spleen, bone marrow, liver and the peritoneal cavity. Because an anti-human SLAMF6 mAb efficiently killed human CLL cells in vitro and in vivo, we propose that a combination of αSlamf6 with ibrutinib should be considered as a novel therapeutic approach for CLL and other B cell tumors. PMID:27029059

  3. A yeast surface display system for the discovery of ligands that trigger cell activation.

    Science.gov (United States)

    Cho, B K; Kieke, M C; Boder, E T; Wittrup, K D; Kranz, D M

    1998-11-01

    Opposing cells often communicate signalling events using multivalent interactions between receptors present on their cell surface. For example, T cells are typically activated when the T cell receptor (TCR) and its associated costimulatory molecules are multivalently engaged by the appropriate ligands present on an antigen presenting cell. In this report, yeast expressing high cell-surface levels of a TCR ligand (a recombinant antibody to the TCR Vbeta domain) were shown to act as 'pseudo' antigen presenting cells and induce T cell activation as monitored by increased levels of CD25 and CD69 and by downregulation of cell surface TCR. Similar levels of T cell activation could occur even when a 30-fold excess of irrelevant yeast was present, suggesting that such a yeast display system, by virtue of its ability to present ligands multivalently, may be used in highly sensitive procedures to identify novel polypeptides that interact multivalently with cell surface receptors and thereby trigger specific cellular responses.

  4. Surface effects after a combination of dental bleaching and enamel microabrasion: An in vitro and in situ study.

    Science.gov (United States)

    Franco, Laura Molinar; Machado, Lucas Silveira; Salomão, Fabio Martins; Dos Santos, Paulo Henrique; Briso, André Luiz Fraga; Sundfeld, Renato Herman

    2016-01-01

    This study evaluated the effects of combining enamel microabrasion and dental bleaching on the physical properties of enamel, using in vitro and in situ conditions and evaluating surface roughness, enamel microhardness and scanning electron microscopy images. One hundred sound bovine teeth were sectioned and cut into discs and randomly divided into 10 study groups (n=10). The results were submitted to Analysis of Variance (ANOVA) for repeated measures, followed by the Tukey test, with significance at 5%. Enamel surface roughness was significantly influenced by microabrasion, regardless of being combined with dental bleaching, for both HS (Human Saliva) or AS (Artificial Saliva) condition. Enamel microhardness was significantly decreased in the groups in which enamel microabrasion was performed, regardless its combination with dental bleaching; although storage in HS reestablished the initial enamel microhardness. It was concluded that dental bleaching does not cause major damage to microabraided enamel, and that only human saliva recovered the initial enamel microhardness. PMID:26830820

  5. Taxol and discodermolide represent a synergistic drug combination in human carcinoma cell lines.

    Science.gov (United States)

    Martello, L A; McDaid, H M; Regl, D L; Yang, C P; Meng, D; Pettus, T R; Kaufman, M D; Arimoto, H; Danishefsky, S J; Smith, A B; Horwitz, S B

    2000-05-01

    Recently, three natural products have been identified, the epothilones, eleutherobin, and discodermolide, whose mechanism of action is similar to that of Taxol in that they stabilize microtubules and block cells in the mitotic phase of the cell cycle. In this report, we have compared and contrasted the effects of these new agents in Taxol-sensitive and -resistant cell lines. We also have taken advantage of a human lung carcinoma cell line, A549-T12, that was isolated as a Taxol-resistant cell line and found to require low concentrations of Taxol (2-6 nM) for normal cell division. This study then examined the ability of these new compounds to substitute for Taxol in sustaining the growth of A549-T12 cells. Immunofluorescence and flow cytometry have both indicated that the epothilones and eleutherobin, but not discodermolide, can substitute for Taxol in this Taxol-dependent cell line. In A549-T12 cells, the presence of Taxol significantly amplified the cytotoxicity of discodermolide, and this phenomenon was not observed in combinations of Taxol with either the epothilones or eleutherobin. Median effect analysis using the combination index method revealed a schedule-independent synergistic interaction between Taxol and discodermolide in four human carcinoma cell lines, an effect that was not observed between Taxol and epothilone B. Flow cytometry revealed that concurrent exposure of A549 cells to Taxol and discodermolide at doses that do not induce mitotic arrest caused an increase in the hypodiploid population, thereby indicating that a possible mechanism for the observed synergy is the potentiation of apoptosis. Our results suggest that Taxol and discodermolide may constitute a promising chemotherapeutic combination.

  6. Toxic effects of xylazine on endothelial cells in combination with cocaine and 6-monoacetylmorphine.

    Science.gov (United States)

    Silva-Torres, L A; Vélez, C; Lyvia Alvarez, J; Ortiz, J G; Zayas, B

    2014-10-01

    The use of xylazine as a drug of abuse has emerged worldwide in the last 7 years, including Puerto Rico. Clinical findings reported that xylazine users present greater physiological deterioration, than heroin users. The aim of this study was to assess the xylazine toxicity on endothelial cells, as this is one of the first tissues impact upon administration. Human umbilical vein endothelial cells in culture were treated with xylazine, cocaine, 6-monoacetylmorphine (heroin metabolite) and its combinations, at concentrations of 0.10-400 μM, for periods of 24, 48 and 72 h. IC50 were calculated and the Annexin V assay implemented to determine the cell death mechanism. Results indicated IC50 values at 24h as follow: xylazine 62 μM, cocaine 210 μM, 6-monoacetylmorphine 300 μM. When these drugs were combined the IC50 value was 57 μM. Annexin V results indicated cell death by an apoptosis mechanism in cells treated with xylazine or in combination. Results demonstrated that xylazine use inhibits the endothelial cell proliferation, at lower concentrations than cocaine and 6-monoacetylmorphine. These findings contribute to the understanding of the toxicity mechanisms induced by xylazine on endothelial cells. PMID:25017475

  7. Effects of recombinant epidermal growth factor receptor antisense adenovirus combined with irradiation on breast cancer cells

    International Nuclear Information System (INIS)

    Objective: To investigate the effects of a recombinant antisense adenovirus for epidermal growth factor receptor (EGFR) combined with irradiation on breast cancer cells. Methods: Human EGFR cDNA fragment was subcloned in the opposite orientation to the cytomegaloviral promoter and inserted into a E1/E3-deleted type 5 adenoviral vector to obtain AdE5 construct which expresses EGFR antisense RNA. Combined with γ-ray irradiation, its effects on clonogenicity and cell cycle phase distribution were studied in a human breast cancer line MDA-MB-23. Results: EGFR protein expression was dramatically inhibited in MDA-MB-231 cells after AdE5 infection. The post-irradiation clonogenicity was reduced by AdE5 in a viral and irradiation dose-dependent manner. Further cytometric analysis showed that AdE5 infection at a MOI of 300 pfu/cell induced a cell cycle progression from radio-resistant G0 + G1 phases to radiosensitive G2 + M phases, resulting in a synergistic effect after combination of these two treatments. Conclusions: The transduction of EGFR antisense RNA by adenoviral vector is effective for antisense strategy targeting EGFR, and increases the cell-killing effect of ionizing radiation on breast cancer cells.(authors)

  8. Large-scale electricity storage utilizing reversible solid oxide cells combined with underground storage of CO2 and CH4

    DEFF Research Database (Denmark)

    Jensen, Søren Højgaard; Graves, Christopher R.; Mogensen, Mogens Bjerg;

    2015-01-01

    for long-term to seasonal periods these technologies are either very expensive or provide a very low round-trip efficiency. Here we describe a novel storage method combining recent advances in reversible solid oxide electrochemical cells with sub-surface storage of CO2 and CH4, thereby enabling large......Electricity storage is needed on an unprecedented scale to sustain the ongoing transition of electricity generation from fossil fuels to intermittent renewable energy sources like wind and solar power. Today pumped hydro is the only commercially viable large-scale electricity storage technology......, but unfortunately it is limited to mountainous regions and therefore difficult to expand. Emerging technologies like adiabatic compressed air energy storage (ACAES) or storage using conventional power-to-gas (P2G) technology combined with underground gas storage can be more widely deployed, but unfortunately...

  9. Bone marrow mesenchymal stem cells combined with minocycline improve spinal cord injury in a rat model

    OpenAIRE

    Chen, Dayong; Zeng, Wei; Fu, Yunfeng; Gao, Meng; Lv, Guohua

    2015-01-01

    The aims of this study were to assess that the effects of bone marrow mesenchymal stem cells (BMSCs) combination with minocycline improve spinal cord injury (SCI) in rat model. In the present study, the Wistar rats were randomly divided into five groups: control group, SCI group, BMSCs group, Minocycline group and BMSCs + minocycline group. Basso, Beattie and Bresnahan (BBB) test and MPO activity were used to assess the effect of combination therapy on locomotion and neutrophil infiltration. ...

  10. Impaired cell surface expression of HLA-B antigens on mesenchymal stem cells and muscle cell progenitors

    DEFF Research Database (Denmark)

    Isa, Adiba; Nehlin, Jan; Sabir, Hardee Jawad;

    2010-01-01

    HLA class-I expression is weak in embryonic stem cells but increases rapidly during lineage progression. It is unknown whether all three classical HLA class-I antigens follow the same developmental program. In the present study, we investigated allele-specific expression of HLA-A, -B, and -C...... at the mRNA and protein levels on human mesenchymal stem cells from bone marrow and adipose tissue as well as striated muscle satellite cells and lymphocytes. Using multicolour flow cytometry, we found high cell surface expression of HLA-A on all stem cells and PBMC examined. Surprisingly, HLA-B was either...... undetectable or very weakly expressed on all stem cells protecting them from complement-dependent cytotoxicity (CDC) using relevant human anti-B and anti-Cw sera. IFNgamma stimulation for 48-72 h was required to induce full HLA-B protein expression. Quantitative real-time RT-PCR showed that IFNgamma induced...

  11. A combined approach for the assessment of cell viability and cell functionality of human fibrochondrocytes for use in tissue engineering.

    Directory of Open Access Journals (Sweden)

    Ingrid Garzón

    Full Text Available Temporo-mandibular joint disc disorders are highly prevalent in adult populations. Autologous chondrocyte implantation is a well-established method for the treatment of several chondral defects. However, very few studies have been carried out using human fibrous chondrocytes from the temporo-mandibular joint (TMJ. One of the main drawbacks associated to chondrocyte cell culture is the possibility that chondrocyte cells kept in culture tend to de-differentiate and to lose cell viability under in in-vitro conditions. In this work, we have isolated human temporo-mandibular joint fibrochondrocytes (TMJF from human disc and we have used a highly-sensitive technique to determine cell viability, cell proliferation and gene expression of nine consecutive cell passages to determine the most appropriate cell passage for use in tissue engineering and future clinical use. Our results revealed that the most potentially viable and functional cell passages were P5-P6, in which an adequate equilibrium between cell viability and the capability to synthesize all major extracellular matrix components exists. The combined action of pro-apoptotic (TRAF5, PHLDA1 and anti-apoptotic genes (SON, HTT, FAIM2 may explain the differential cell viability levels that we found in this study. These results suggest that TMJF should be used at P5-P6 for cell therapy protocols.

  12. High quality crystalline silicon surface passivation by combined intrinsic and n-type hydrogenated amorphous silicon

    NARCIS (Netherlands)

    Schuttauf, J.A.; van der Werf, C.H.M.; Kielen, I.M.; van Sark, W.G.J.H.M.; Rath, J.K.

    2011-01-01

    We investigate the influence of thermal annealing on the passivation quality of crystalline silicon (c-Si) surfaces by intrinsic and n-type hydrogenated amorphous silicon (a-Si:H) films. For temperatures up to 255 C, we find an increase in surface passivation quality, corresponding to a decreased da

  13. Surface Modification by Means of Laser Melting Combined with Shot Peening : A Novel Approach

    NARCIS (Netherlands)

    Noordhuis, J.; Hosson, J.Th.M. De

    1992-01-01

    Among the available laser applications laser surface melting has turned out to be a powerful technique for the production of wear-resistant layers. Despite the advantages of this process, laser surface melting results in tensile stresses which may assist crack propagation. In this paper it will be s

  14. SURFACE MODIFICATION BY MEANS OF LASER MELTING COMBINED WITH SHOT PEENING - A NOVEL-APPROACH

    NARCIS (Netherlands)

    NOORDHUIS, J; DEHOSSON, JTM

    1992-01-01

    Among the available laser applications laser surface melting has turned out to be a powerful technique for the production of wear-resistant layers. Despite the advantages of this process, laser surface melting results in tensile stresses which may assist crack propagation. In this paper it will be s

  15. A simplified model for dynamics of cell rolling and cell-surface adhesion

    Energy Technology Data Exchange (ETDEWEB)

    Cimrák, Ivan, E-mail: ivan.cimrak@fri.uniza.sk [Cell-in-fluid Research Group, http://cell-in-fluid.fri.uniza.sk Faculty of Management Science and Informatics, University of Žilina Univerzitná 8215/1, 010 26 Žilina (Slovakia)

    2015-03-10

    We propose a three dimensional model for the adhesion and rolling of biological cells on surfaces. We study cells moving in shear flow above a wall to which they can adhere via specific receptor-ligand bonds based on receptors from selectin as well as integrin family. The computational fluid dynamics are governed by the lattice-Boltzmann method. The movement and the deformation of the cells is described by the immersed boundary method. Both methods are fully coupled by implementing a two-way fluid-structure interaction. The adhesion mechanism is modelled by adhesive bonds including stochastic rules for their creation and rupture. We explore a simplified model with dissociation rate independent of the length of the bonds. We demonstrate that this model is able to resemble the mesoscopic properties, such as velocity of rolling cells.

  16. Thermodynamic optimization of solid oxide fuel cell based combined cycle cogeneration plant

    Energy Technology Data Exchange (ETDEWEB)

    Odukoya, A.; Reddy, B.V. [University of Ontario Inst. of Technology, Oshawa, ON (Canada). Dept. of Mechanical Engineering; Carretero, J.A. [New Brunswick Univ., Fredericton, NB (Canada). Dept. of Mechanical Engineering

    2010-07-01

    Although coal has the highest greenhouse gas emission of all fossil fuels it is the most abundant of all the fuels. Optimization of new and existing power plant designs will help increase the operational efficiency of power generation. In particular, there is a need to optimize the design and operating parameters of power plants using integrated gasification combined cycle cogeneration technology. This study investigated the optimal operating condition of a co-fired combined cycle cogeneration power plant with solid oxide fuel cell (SOFC) combination fuelled with coal and natural gas. It examined the macroscopic detail of the plant while optimizing the operating parameters of individual components such as the gasifier, the consumption of methane and carbon monoxide in the fuel cell and the consumption of fuel in the combustion chamber of the gas turbine. The optimization of the entire plant was used to determine the best mode of operating the plant for a set of conditions within suggested limits. The study also found efficient ways to perform iterative processes to find exit conditions from the gasifier, fuel cell, gas turbine combustion chamber and exit condition from the gas turbine. The maximum fuel cell net work output, combined cycle net work output, combined cycle thermal efficiency and cogeneration efficiency were determined. The optimal pressure ratio, temperature of operation of the SOFC and, gas turbine inlet temperature were determined using a sequential quadratic program solver based on the Quasi-Newton algorithm. 18 refs., 5 tabs., 2 figs.

  17. Micropatterned polysaccharide surfaces via laser ablation for cell guidance

    Energy Technology Data Exchange (ETDEWEB)

    Barbucci, Rolando; Lamponi, Stefania; Pasqui, Daniela; Rossi, Antonella; Weber, Elisabetta

    2003-03-03

    Micropatterned materials were obtained by a controlled laser ablation of a photoimmobilised homogeneous layer of hyaluronic acid (Hyal) and its sulphated derivative (HyalS). The photoimmobilisation was performed by coating the polysaccharide, adequately functionalised with a photoreactive group, on aminosilanised glass substrate and immobilising it on the surface under UV light. Hyal or HyalS photoimmobilised samples were then subjected to laser ablation with wavelengths in the UV regions in order to drill the pattern. Four different patterns with stripes of 100, 50, 25 and 10 {mu}m were generated. A chemical characterisation by attenuated total reflection/Fourier transform infrared (ATR/FT-IR) and time of flight-secondary ions mass spectrometry (TOF-SIMS) confirmed the success of the laser ablation procedure and the presence of alternating stripes of polysaccharide and native glass. The exact dimensions of the stripes were determined by atomic force microscopy. The analysis of cell behaviour in terms of adhesion, proliferation and movement using mouse fibroblasts (3T3 line) and bovine aortic endothelial cells (BAEC) was also performed.

  18. EXAFS Study of Uranyl Complexation at Pseudomonas fluorescens Cell Surfaces

    Science.gov (United States)

    Bencheikh, R.; Bargar, J. R.; Tebo, B. M.

    2002-12-01

    Little is known about the roles of microbial biomass as a sink and source for uranium in contaminated aquifers, nor of the impact of bacterial biochemistry on uranium speciation in the subsurface. A significant role is implied by the high affinities of both Gram positive and Gram negative cells for binding uranyl (UO2{ 2+}). In the present study, Extended X-ray Absorption Fine Structure (EXAFS) spectroscopy was used to identify membrane functional groups involved in uranyl binding to the Gram negative bacterium Pseudomonas fluorescens from pH 3 to pH 8. Throughout this pH-range, EXAFS spectra can be described primarily in terms of coordination of carboxylic groups to uranyl. U-C distances characteristic of 4-, 5- and 8- membered rings were observed, as well as the possibility of phosphato groups. Both shell-by-shell fits and principle component analyses indicate that the functional groups involved in binding of uranyl to the cell surface do not vary systematically across the pH range investigated. This result contrasts with EXAFS results of uranyl sorbed to Gram positive bacteria, and suggests an important role for long-chain carboxylate-terminated membrane functional groups in binding uranyl.

  19. Effects of DNP on the cell surface properties of marine bacteria and its implication for adhesion to surfaces

    Digital Repository Service at National Institute of Oceanography (India)

    Jain, A.; Nishad, K.K.; Bhosle, N.B.

    The effect of 2, 4-dinitrophenol (DNP) on extracelluar polysaccharides (EPS), cell surface charge, and hydrophobicity of six marine bacterial cultures was studied, and its influence on attachment of these bacteria to glass and polystyrene...

  20. The combination effects of bendamustine with antimetabolites against childhood acute lymphoblastic leukemia cells.

    Science.gov (United States)

    Goto, Shoko; Goto, Hiroaki; Yokosuka, Tomoko

    2016-05-01

    Bendamustine combined with other drugs is clinically efficacious for some adult lymphoid malignancies, but to date there are no reports of the use of such combinatorial approaches in pediatric patients. We investigated the in vitro activity of bendamustine combined with other antimetabolite drugs on B cell precursor acute lymphoblastic leukemia (BCP-ALL) cell lines established from pediatric patients with refractory or relapsed ALL. We also developed a mathematically drown improved isobologram method to assess the data objectively. Three BCP-ALL cell lines; YCUB-2, YCUB-5, and YCUB-6, were simultaneously exposed to various concentrations of bendamustine and cladribine, cytarabine, fludarabine, or clofarabine. Cell growth inhibition was determined using the WST-8 assay. Combinatorial effects were estimated using our improved isobologram method with IC80 (drug concentration corresponding to 80 % of maximum inhibition). Bendamustine alone inhibited ALL cell growth with mean IC80 values of 11.30-18.90 μg/ml. Combinations of bendamustine with other drugs produced the following effects: (1) cladribine; synergistic-to-additive on all cell lines; (2) cytarabine; synergistic-to-additive on YCUB-5 and YCUB-6, and synergistic-to-antagonistic on YCUB-2; (3) fludarabine; additive-to-antagonistic on YCUB-5, and synergistic-to-antagonistic on YCUB-2 and YCUB-6; (4) clofarabine; additive-to-antagonistic on all cell lines. Flow cytometric analysis also showed the combination effects of bendamustine and cladribine. Bendamustine/cladribine or bendamustine/cytarabine may thus represent a promising combination for salvage treatment in childhood ALL. PMID:26886449

  1. Human epithelial cells exposed to functionalized multiwalled carbon nanotubes: interactions and cell surface modifications.

    Science.gov (United States)

    Fanizza, C; Casciardi, S; Incoronato, F; Cavallo, D; Ursini, C L; Ciervo, A; Maiello, R; Fresegna, A M; Marcelloni, A M; Lega, D; Alvino, A; Baiguera, S

    2015-09-01

    With the expansion of the production and applications of multiwalled carbon nanotubes (MWCNTs) in several industrial and science branches, the potential adverse effects on human health have attracted attention. Numerous studies have been conducted to evaluate how chemical functionalization may affect MWCNT effects; however, controversial data have been reported, showing either increased or reduced toxicity. In particular, the impact of carboxylation on MWCNT cytotoxicity is far from being completely understood. The aim of this work was the evaluation of the modifications induced by carboxylated-MWCNTs (MWCNTs-COOH) on cell surface and the study of cell-MWCNT-COOH interactions by means of field emission scanning electron microscope (FESEM). Human pulmonary epithelial cells (A549) were incubated with MWCNTs-COOH for different exposure times and concentrations (10 μg/mL for 1, 2, 4 h; 5, 10, 20 μg/mL for 24 h). At short incubation time, MWCNTs-COOH were easily observed associated with plasma membrane and in contact with microvilli. After 24 h exposure, FESEM analysis revealed that MWCNTs-COOH induced evident changes in the cellular surface in comparison to control cells: treated cells showed blebs, holes and a depletion of the microvilli density in association with structure modifications, such as widening and/or lengthening. In particular, an increase of cells showing holes and microvilli structure alterations was observed at 20 μg/mL concentration. FESEM analysis showed nanotube agglomerates, of different sizes, entering into the cell with two different mechanisms: inward bending of the membrane followed by nanotube sinking, and nanotube internalization directly through holes. The observed morphological microvilli modifications, induced by MWCNTs-COOH, could affect epithelial functions, such as the control of surfactant production and secretion, leading to pathological conditions, such as alveolar proteinosis. More detailed studies will be, however, necessary to

  2. Synthesis and characterization of hybrid micro/nano-structured NiTi surfaces by a combination of etching and anodizing

    International Nuclear Information System (INIS)

    The purpose of this study was to generate hybrid micro/nano-structures on biomedical nickel–titanium alloy (NiTi). To achieve this, NiTi surfaces were firstly electrochemically etched and then anodized in fluoride-containing electrolyte. With the etching process, the NiTi surface was micro-roughened through the formation of micropits uniformly distributed over the entire surface. Following the subsequent anodizing process, self-organized nanotube structures enriched in TiO2 could be superimposed on the etched surface under specific conditions. Furthermore, the anodizing treatment significantly reduced water contact angles and increased the surface free energy compared to the surfaces prior to anodizing. The results of this study show for the first time that it is possible to create hybrid micro/nano-structures on biomedical NiTi alloys by combining electrochemical etching and anodizing under controlled conditions. These novel structures are expected to significantly enhance the surface biofunctionality of the material when compared to conventional implant devices with either micro- or nano-structured surfaces. (paper)

  3. Synthesis and characterization of hybrid micro/nano-structured NiTi surfaces by a combination of etching and anodizing

    Science.gov (United States)

    Huan, Z.; Fratila-Apachitei, L. E.; Apachitei, I.; Duszczyk, J.

    2014-02-01

    The purpose of this study was to generate hybrid micro/nano-structures on biomedical nickel-titanium alloy (NiTi). To achieve this, NiTi surfaces were firstly electrochemically etched and then anodized in fluoride-containing electrolyte. With the etching process, the NiTi surface was micro-roughened through the formation of micropits uniformly distributed over the entire surface. Following the subsequent anodizing process, self-organized nanotube structures enriched in TiO2 could be superimposed on the etched surface under specific conditions. Furthermore, the anodizing treatment significantly reduced water contact angles and increased the surface free energy compared to the surfaces prior to anodizing. The results of this study show for the first time that it is possible to create hybrid micro/nano-structures on biomedical NiTi alloys by combining electrochemical etching and anodizing under controlled conditions. These novel structures are expected to significantly enhance the surface biofunctionality of the material when compared to conventional implant devices with either micro- or nano-structured surfaces.

  4. Micro-tubular flame-assisted fuel cells for micro-combined heat and power systems

    Science.gov (United States)

    Milcarek, Ryan J.; Wang, Kang; Falkenstein-Smith, Ryan L.; Ahn, Jeongmin

    2016-02-01

    Currently the role of fuel cells in future power generation is being examined, tested and discussed. However, implementing systems is more difficult because of sealing challenges, slow start-up and complex thermal management and fuel processing. A novel furnace system with a flame-assisted fuel cell is proposed that combines the thermal management and fuel processing systems by utilizing fuel-rich combustion. In addition, the flame-assisted fuel cell furnace is a micro-combined heat and power system, which can produce electricity for homes or businesses, providing resilience during power disruption while still providing heat. A micro-tubular solid oxide fuel cell achieves a significant performance of 430 mW cm-2 operating in a model fuel-rich exhaust stream.

  5. Effects of IL-24 gene combined with ionizing radiaiton on apoptosis in PC-3 cell line

    International Nuclear Information System (INIS)

    Objective: To study the effects of IL-24 gene combined with ionizing radiation on apoptosis in PC-3 cell line in order to prepare the ground for combined therapy of IL-24 gene and ionzing radiation for tumor. Methods: The experiment was divided into sham irradiation group and irradiation groups with different irradiation doses, which were 2, 4, 6, 8, 12 and 18 Gy, respectively. To detect the expression of IL-24 gene, three groups were included, which were control group (1 x PBS), vector group and IL-24 gene group. To detect the apoptotic effect of IL-24 gene combined with ionization radiation on PC-3 cell line, the experiment was divided into control group, vector group, IL-24 gene group, irradiation group (6 Gy), vector combined with irradiation group and IL-24 gene combined with irradiation group. Alkaline lysis assay was used to extract and purify the plasmid. Plasmids were transfected into PC-3 cell line by polyethyleneimine (PEI) in vitro. The expression of the interest gene was detected by RT-PCR. The changes of apoptosis in PC-3 cell line were detected by flow cytometry (FCM) using the staining of Annexin-V and PI. Results: Compared with sham-irradiation group, the apoptotic percentage of PC-3 cell line did not show marked change after 2 and 4 Gy X-rays irradiation for 48 h (P>0.05). The apoptotic percentage was increased significantly after X-rays irradiation with the dose of 6 Gy (P<0.01), and the mean apoptotic percentage of PC-3 cell line was increased by a factor of 1.6 to 3.0 compared with sham-irradiation group. The expression of IL-24 gene could be observed in PC-3 cell line transfected by IL-24 gene except in control group and vector group, and all of them showed the expression of GAPDH gene. As compared with the other groups, the number of early apoptotic cells of PC-3 cell line in the IL-24 gene combined with irradiation group was increased significantly (P<0.05) except in irradiation group. The number of late apoptotic and necrotic cells of PC-3

  6. Campylobacter jejuni cocultured with epithelial cells reduces surface capsular polysaccharide expression.

    LENUS (Irish Health Repository)

    Corcionivoschi, N

    2012-02-01

    The host cell environment can alter bacterial pathogenicity. We employed a combination of cellular and molecular techniques to study the expression of Campylobacter jejuni polysaccharides cocultured with HCT-8 epithelial cells. After two passages, the amount of membrane-bound high-molecular-weight polysaccharide was considerably reduced. Microarray profiling confirmed significant downregulation of capsular polysaccharide (CPS) locus genes. Experiments using conditioned media showed that sugar depletion occurred only when the bacterial and epithelial cells were cocultured. CPS depletion occurred when C. jejuni organisms were exposed to conditioned media from a different C. jejuni strain but not when exposed to conditioned media from other bacterial species. Proteinase K or heat treatment of conditioned media under coculture conditions abrogated the effect on the sugars, as did formaldehyde fixation and cycloheximide treatment of host cells or chloramphenicol treatment of the bacteria. However, sugar depletion was not affected in flagellar export (fliQ) and quorum-sensing (luxS) gene mutants. Passaged C. jejuni showed reduced invasiveness and increased serum sensitivity in vitro. C. jejuni alters its surface polysaccharides when cocultured with epithelial cells, suggesting the existence of a cross talk mechanism that modulates CPS expression during infection.

  7. Surface plasma resonance enhanced photocurrent generation in NiO photoanode based solar cells

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Zhong; Cui, Jin [Michael Grätzel Center for Mesoscopic Solar Cells, Wuhan National Laboratory for Optoelectronics Department, Huazhong University of Science and Technology, 1037 Luoyu Road, Wuhan 430074, Hubei (China); Li, Junpeng [State Key Laboratory of Advanced Technologies for Comprehensive Utilization of Platinum Metals, Kunming Institute of Precious Metals, Kunming 650106 (China); Cao, Kun; Yuan, Shuai [Michael Grätzel Center for Mesoscopic Solar Cells, Wuhan National Laboratory for Optoelectronics Department, Huazhong University of Science and Technology, 1037 Luoyu Road, Wuhan 430074, Hubei (China); Cheng, Yibing [Michael Grätzel Center for Mesoscopic Solar Cells, Wuhan National Laboratory for Optoelectronics Department, Huazhong University of Science and Technology, 1037 Luoyu Road, Wuhan 430074, Hubei (China); Department of Materials Engineering, Monash University, Melbourne, Victoria 3800 (Australia); Wang, Mingkui, E-mail: mingkui.wang@mail.hust.edu.cn [Michael Grätzel Center for Mesoscopic Solar Cells, Wuhan National Laboratory for Optoelectronics Department, Huazhong University of Science and Technology, 1037 Luoyu Road, Wuhan 430074, Hubei (China)

    2015-09-15

    Highlights: • SPR effect from Au-nanostructures was first investigated in NiO-based solar cells. • Enhanced photocurrent generation was observed in p-DSC and perovskite solar cell. • Au-nanorods SPR effect induced charge kinetics were investigated. - Abstract: Surface plasma resonance (SPR) effect has been demonstrated to improve solar cell performance. This work reports on the SPR effect from Au nanorod@SiO{sub 2} on p-type dye-sensitized solar cells. Au nanorod@SiO{sub 2} works as an antenna to transform photons with long wavelength into electric field followed by an enhanced excitation of dye. The devices using the NiO electrode containing Au nanorod@SiO{sub 2} shows overall power conversion efficiencies of about 0.2% in combination with I{sup −}/I{sub 3}{sup −} electrolyte, and 0.29% with T{sup −}/T{sub 2} electrolyte, which are superior to those without adding Au nanorods. Detailed investigation including spectroscopy and transient photovoltage decay measurements reveals that plasma effect of Au nanorod@SiO{sub 2} contribute to charge injection efficiency, and thus on the photocurrent. The effect of Au NRs can be further extended to the inverted planar perovskite solar cells, showing obviously improvement in photocurrent.

  8. Prospective Isolation of Murine and Human Bone Marrow Mesenchymal Stem Cells Based on Surface Markers

    Directory of Open Access Journals (Sweden)

    Yo Mabuchi

    2013-01-01

    Full Text Available Mesenchymal stem cells (MSCs are currently defined as multipotent stromal cells that undergo sustained in vitro growth and can give rise to cells of multiple mesenchymal lineages, such as adipocytes, chondrocytes, and osteoblasts. The regenerative and immunosuppressive properties of MSCs have led to numerous clinical trials exploring their utility for the treatment of a variety of diseases (e.g., acute graft-versus-host disease, Crohn’s disease, multiple sclerosis, osteoarthritis, and cardiovascular diseases including heart failure and myocardial infarction. On the other hand, conventionally cultured MSCs reflect heterogeneous populations that often contain contaminating cells due to the significant variability in isolation methods and the lack of specific MSC markers. This review article focuses on recent developments in the MSC research field, with a special emphasis on the identification of novel surface markers for the in vivo localization and prospective isolation of murine and human MSCs. Furthermore, we discuss the physiological importance of MSC subtypes in vivo with specific reference to data supporting their contribution to HSC niche homeostasis. The isolation of MSCs using selective markers (combination of PDGFRα and Sca-1 is crucial to address the many unanswered questions pertaining to these cells and has the potential to enhance their therapeutic potential enormously.

  9. Surface plasma resonance enhanced photocurrent generation in NiO photoanode based solar cells

    International Nuclear Information System (INIS)

    Highlights: • SPR effect from Au-nanostructures was first investigated in NiO-based solar cells. • Enhanced photocurrent generation was observed in p-DSC and perovskite solar cell. • Au-nanorods SPR effect induced charge kinetics were investigated. - Abstract: Surface plasma resonance (SPR) effect has been demonstrated to improve solar cell performance. This work reports on the SPR effect from Au nanorod@SiO2 on p-type dye-sensitized solar cells. Au nanorod@SiO2 works as an antenna to transform photons with long wavelength into electric field followed by an enhanced excitation of dye. The devices using the NiO electrode containing Au nanorod@SiO2 shows overall power conversion efficiencies of about 0.2% in combination with I−/I3− electrolyte, and 0.29% with T−/T2 electrolyte, which are superior to those without adding Au nanorods. Detailed investigation including spectroscopy and transient photovoltage decay measurements reveals that plasma effect of Au nanorod@SiO2 contribute to charge injection efficiency, and thus on the photocurrent. The effect of Au NRs can be further extended to the inverted planar perovskite solar cells, showing obviously improvement in photocurrent

  10. Cell adhesion on Ti surface with controlled roughness

    Directory of Open Access Journals (Sweden)

    Burgos-Asperilla, Laura

    2015-06-01

    Full Text Available In this report, the in situ interaction between Saos-2 osteoblast cells and a smooth Ti surface was examined over time. The adhesion kinetics and mechanisms of cellular proliferation were monitored by quartz crystal microbalance (QCM and electrochemical impedance spectroscopy (EIS. The rate of Saos-2 attachment on Ti surfaces, obtained from the measurements performed with the QCM, is a first-order reaction, with k=2.10−3 min−1. The impedance measurements indicate that in the absence of cells, the Ti resistance diminishes over time (7 days, due to the presence of amino acids and proteins from the culture medium that have been a dsorbed, while in the presence of osteoblasts, this decrease is much greater because of the compounds generated by the cells that accelerate the dissolution of Ti.En este trabajo, se ha estudiado la interacción in situ entre células osteoblásticas Saos-2 y una superficie de Ti de rugosidad controlada a lo largo del tiempo. El estudio de la cinética y los mecanismos de proliferación celular de adhesión se ha realizado a través de la microbalanza de cristal de cuarzo (QCM y espectroscopía de impedancia electroquímica (EIS. La velocidad de adhesión de los osteoblastos sobre la superficie de Ti obtenida a través de medidas con la QCM, sigue una reacción de primer orden, con k=2×10−3 min−1. Los ensayos de impedancia indican que, en ausencia de las células, la resistencia del Ti disminuye con el tiempo (7 días, debido a la presencia de aminoácidos y proteínas del medio de cultivo que se han adsorbido, mientras que en presencia de células, esta disminución es mucho mayor debido a los productos metabólicos generados por las células que aceleran la disolución del Ti.

  11. Aclidinium bromide combined with formoterol inhibits remodeling parameters in lung epithelial cells through cAMP.

    Science.gov (United States)

    Lambers, Christopher; Costa, Luigi; Ying, Qi; Zhong, Jun; Lardinois, Didier; Dekan, Gerhard; Schuller, Elisabeth; Roth, Michael

    2015-12-01

    Combined muscarinic receptor antagonists and long acting β2-agonists improve symptom control in chronic obstructive pulmonary disease (COPD) significantly. In clinical studies aclidinium bromide achieved better beneficial effects than other bronchodilators; however, the underlying molecular mechanisms are unknown. This study assessed the effect of aclidinium bromide combined with formoterol on COPD lung (n=20) and non-COPD lung (n=10) derived epithelial cells stimulated with TGF-β1+carbachol on: (i) the generation of mesenchymal cells in relation to epithelial cells, (II) extracellular matrix (ECM) deposition, and (iii) the interaction of ECM on the generation of epithelial and mesenchymal cells. TGF-β1+carbachol enhanced the generation of mesenchymal cells, which was significantly reduced by aclidinium bromide or formoterol. The effect of combined drugs was additive. Inhibition of p38 MAP kinase and Smad by specific inhibitors or aclidinium bromide reduced the generation of mesenchymal cells. In mesenchymal cells, TGF-β1+carbachol induced the deposition of collagen-I and fibronectin which was prevented by both drugs dose-dependently. Formoterol alone reduced collagen-I deposition via cAMP, this however, was overruled by TGF-β1+carbachol and rescued by aclidinium bromide. Inhibition of fibronectin was cAMP independent, but involved p38 MAP kinase and Smad. Seeding epithelial cells on ECM collagen-I and fibronectin induced mesenchymal cell generation, which was reduced by aclidinium bromide and formoterol. Our results suggest that the beneficial effect of aclidinium bromide and formoterol involves cAMP affecting both, the accumulation of mesenchymal cells and ECM remodeling, which may explain the beneficial effect of the drugs on lung function in COPD. PMID:26546746

  12. The structure and assembly of surface layer proteins : a combined approach of in silico and experimental methods

    International Nuclear Information System (INIS)

    Self-assembly of matter is one of nature's most sophisticated strategies to organize molecules on a large scale and to create order from disorder. Surface (S-)layer proteins self-assemble in a highly reproducible and robust fashion in order to form crystalline layers that completely cover and protect prokaryotic cells. Long conserved during evolution, S-layers constitute a unique model system to study the molecular mechanisms of functional self-assembly, while additionally, they provide a basic matrix for the specific construction of ordered nanostructures. Due to their intrinsic capabilities to self-assemble into two-dimensional crystals, the elucidation of the three-dimensional structure of single S-layer proteins demands an approach beyond conventional structure determination methods. In this work, computer simulations were combined with experimental techniques in order to study the structure and intra- and intermolecular potentials guiding the proteins to self-assemble into lattices with different symmetries. Molecular dynamics, Monte Carlo methods, small-angle X-ray scattering involving a new theoretical description, and AFM-based single-molecule force spectroscopy yield new insights into the three-dimensional structure of S-layer proteins, the location, type and distribution of amino acids in S-layer lattices, the molecular mechanisms behind the self-assembly process, the mechanical stability and adaptive structural conformations that S-layer proteins are able to establish. In silico studies - embedded in an adequate experimental and theoretical scaffold - offer the possibility to calculate structural and thermodynamic features of proteins, while this work demonstrates the growing impact of such theoretical techniques in the fascinating field of biophysics at the nano-scale. (author)

  13. Cell-surface expression of Hsp70 on hematopoietic cancer cells after inhibition of HDAC activity

    DEFF Research Database (Denmark)

    Jensen, Helle

    The function of Hsp70 depends on its cellular location: When located intracellularly it exerts cytoprotective and anti-apoptotic functions, whereas it exerts immunostimulatory functions when located extracellularly. Secreted Hsp70 is for example involved in cross-presentation of cancer-derived an......The function of Hsp70 depends on its cellular location: When located intracellularly it exerts cytoprotective and anti-apoptotic functions, whereas it exerts immunostimulatory functions when located extracellularly. Secreted Hsp70 is for example involved in cross-presentation of cancer......-derived antigenic peptides, a function which is currently explored in immunotherapeutic approaches against cancer. Additionally, membrane-bound Hsp70 can stimulate antigen presenting cells to release proinflammatory cytokines and can provide a target structure for NK cell-mediated lysis. Human cancer cells...... frequently express Hsp70 on their cell surface, whereas the corresponding normal tissues do not. In addition, several clinically applied reagents, such as alkyl-lysophospholipides, chemotherapeutic agents, and anti-inflammatory reagents, have been found to enhance Hsp70 cell surface expression on cancer...

  14. Engineering of silicon surfaces at the micro- and nanoscales for cell adhesion and migration control

    Directory of Open Access Journals (Sweden)

    Torres-Costa V

    2012-02-01

    Full Text Available Vicente Torres-Costa1, Gonzalo Martínez-Muñoz2, Vanessa Sánchez-Vaquero3, Álvaro Muñoz-Noval1, Laura González-Méndez3, Esther Punzón-Quijorna1,4, Darío Gallach-Pérez1, Miguel Manso-Silván1, Aurelio Climent-Font1,4, Josefa P García-Ruiz3, Raúl J Martín-Palma11Department of Applied Physics, 2Department of Computer Science, 3Department of Molecular Biology, 4Centre for Micro Analysis of Materials, Universidad Autónoma de Madrid, Madrid, SpainAbstract: The engineering of surface patterns is a powerful tool for analyzing cellular communication factors involved in the processes of adhesion, migration, and expansion, which can have a notable impact on therapeutic applications including tissue engineering. In this regard, the main objective of this research was to fabricate patterned and textured surfaces at micron- and nanoscale levels, respectively, with very different chemical and topographic characteristics to control cell–substrate interactions. For this task, one-dimensional (1-D and two-dimensional (2-D patterns combining silicon and nanostructured porous silicon were engineered by ion beam irradiation and subsequent electrochemical etch. The experimental results show that under the influence of chemical and morphological stimuli, human mesenchymal stem cells polarize and move directionally toward or away from the particular stimulus. Furthermore, a computational model was developed aiming at understanding cell behavior by reproducing the surface distribution and migration of human mesenchymal stem cells observed experimentally.Keywords: surface patterns, silicon, hMSCs, ion-beam patterning

  15. Corner detection in color images by multiscale combination of end-stopped cortical cells.

    OpenAIRE

    Würtz, R.P.; Lourens, T.

    1997-01-01

    We present a corner-detection algorithm based on a model for end-stopping cells in the visual cortex. Shortcomings of this model are overcome by a combination over several scales. The notion of an end-stopped cell and the resulting corner detector is generalized to color channels in a biologically plausible way. The resulting corner detection method yields good results in the presence of high frequency texture, noise, varying contrast, and rounded corners. This compares favorably with known c...

  16. Efficacy of gemcitabine and cetuximab combination treatment in head and neck squamous cell carcinoma

    OpenAIRE

    MASEKI, SHINICHIRO; IJICHI, KEI; NAKANISHI, HAYAO; Hasegawa, Yasuhisa; OGAWA, TETSUYA; Murakami, Shingo

    2013-01-01

    Head and neck squamous cell carcinoma (HNSCC) may be curable with surgery, radiation and chemotherapy in its early stages. However, recurrence and metastasis often prevail following primary treatment in advanced stage cases and are associated with significant morbidity and mortality. In this study we investigated the combination therapy of gemcitabine and cetuximab for HNSCC. The UM-SCC-6 and UM-SCC-23 HNSCC cell lines were analyzed following treatment with gemcitabine and cetuximab. To deter...

  17. Autologous Bone Marrow Stem Cells combined with Allograft Cancellous Bone in Treatment of Nonunion

    OpenAIRE

    Le Thua Trung Hau; Duc Phu Bui; Nguyen Duy Thang; Pham Dang Nhat; Le Quy Bao; Nguyen Phan Huy; Tran Ngoc Vu; Le Phuoc Quang; Boeckx willy Denis; Mey Albert De

    2015-01-01

    Autologous cancellous bone graft is currently used as a gold standard method for treatment of bone nonunion. However, there is a limit to the amount of autologous cancellous bone that can be harvested and the donor site morbidity presents a major disadvantage to autologous bone grafting. Embedding viable cells within biological scaffolds appears to be extremely promising. The purpose of this study was to assess the outcome of autologous bone marrow stem cells combined with a cancellous bone a...

  18. Discovery of a low order drug-cell response surface for applications in personalized medicine

    International Nuclear Information System (INIS)

    The cell is a complex system involving numerous components, which may often interact in a non-linear dynamic manner. Diseases at the cellular level are thus likely to involve multiple cellular constituents and pathways. As some drugs, or drug combinations, may act synergistically on these multiple pathways, they might be more effective than the respective single target agents. Optimizing a drug mixture for a given disease in a particular patient is particularly challenging due to both the difficulty in the selection of the drug mixture components to start out with, and the all-important doses of these drugs to be applied. For n concentrations of m drugs, in principle, nm combinations will have to be tested. As this may lead to a costly and time-consuming investigation for each individual patient, we have developed a Feedback System Control (FSC) technique which can rapidly select the optimal drug–dose combination from the often millions of possible combinations. By testing this FSC technique in a number of experimental systems representing different disease states, we found that the response of cells to multiple drugs is well described by a low order, rather smooth, drug-mixture-input/drug-effect-output multidimensional surface. The main consequences of this are that optimal drug combinations can be found in a surprisingly small number of tests, and that translation from in vitro to in vivo is simplified. This points to the possibility of personalized optimal drug mixtures in the near future. This unexpectedly simple input–output relationship may also lead to a simple solution for handling the issue of human diversity in cancer therapeutics. (paper)

  19. X-ray photoelectron spectroscopy for the study of microbial cell surfaces

    NARCIS (Netherlands)

    van der Mei, Henderina C; de Vries, Jacob; Busscher, Hendrik J

    2000-01-01

    X-ray photoelectron spectroscopy (XPS) is well known for the characterisation of material surfaces, but at first glance, is an unexpected technique to study the composition of microbial cell surfaces. Despite the fact that intimate contact between materials and microbial cell surfaces occurs in many

  20. Surface Grafted Glycopolymer Brushes to Enhance Selective Adhesion of HepG2 Cells

    DEFF Research Database (Denmark)

    Chernyy, Sergey; Jensen, Bettina Elisabeth Brøgger; Shimizu, Kyoko;

    2013-01-01

    of the cell periphery. On the other hand the cells on bare glass substrate display spheroid morphology. Further analysis using ToF-SIMS imaging shows that the HepG2 cells on glycopolymer surfaces is enriched with protein fragment along the cell periphery which is absent in the case of cells on bare glass...

  1. Definitive Endoderm Differentiation of Human Embryonic Stem Cells Combined with Selective Elimination of Undifferentiated Cells by Methionine Deprivation.

    Science.gov (United States)

    Tsuyama, Tomonori; Shiraki, Nobuaki; Kume, Shoen

    2016-01-01

    Human embryonic stem cells (ESCs) show a characteristic feature in that they are highly dependent on methionine metabolism. Undifferentiated human ESCs cannot survive under condition that methionine is deprived from culture medium. We describe here a procedure for definitive endoderm differentiation from human ESCs, in which human ESCs are subject to 10 days' (d) differentiation combined with methionine deprivation between differentiation days (d) 8 to (d) 10. Methionine deprivation results in elimination of undifferentiated cells from the culture with no significant loss of definitive endoderm cells, as compared to those cultured under complete condition throughout the whole culture period.

  2. Aged mice have increased inflammatory monocyte concentration and altered expression of cell-surface functional receptors

    Indian Academy of Sciences (India)

    Kelley Strohacker; Whitney L Breslin; Katie C Carpenter; Brian K McFarlin

    2012-03-01

    The expression of monocyte cell-surface receptors represents one index of immune dysfunction, which is common with aging. Although mouse models of aging are prevalent, monocyte subset assessment is rare. Our purpose was to compare cell receptor expression on classic (CD115+/Gr-1high) and non-classic (CD115+/Gr-1low) monocytes from 80- or 20-week-old CD-1 mice. Three-colour flow cytometry was used to determine the concentration of monocyte subsets and their respective cell-surface expression of TLR2, TLR4, CD80, CD86, MHC II and CD54. These receptors were selected because they have been previously associated with altered monocyte function. Data were analysed with independent -tests; significance was set at < 0.05. Old mice had a greater concentration of both classic (258%, =0.003) and non-classic (70%, =0.026) monocytes. The classic : non-classic monocyte ratio doubled in old as compared with that in young mice (=0.006), indicating a pro-inflammatory shift. TLR4 ($\\downarrow$27%, =0.001) and CD80 ($\\downarrow$37%, =0.004) were decreased on classic monocytes from old as compared with those from young mice. TLR2 ($\\uparrow$24%, =0.002) and MHCII ($\\downarrow$21%, =0.026) were altered on non-classic monocytes from old as compared with those from young mice. The increased classic : non-classic monocyte ratio combined with changes in the cell-surface receptor expression on both monocyte subsets is indicative of immune dysfunction, which may increase age-associated disease risk.

  3. Surface engineering and characterization of quantum dots and its application in living cell imaging

    Directory of Open Access Journals (Sweden)

    Han QIU

    2016-06-01

    Full Text Available Objective  Surface modification of hydrophobic nanoparticles is a key process for their application in the biological medicine fields. The aim of present study is to prepare the immunofluorescent probes by conjugation of hydrophilic surface-engineered quantum dots (QDs and monoclonal antibody for use of fluorescence labeled cells tracing. Methods  The bovine serum albumin (BSA with excellent water-solubility and biocompatibility was employed as the emulsifying agent, and used for surface modification of hydrophobic QDs under ultrasonication. The diameter, fluorescence spectrum and cytotoxicity of BSA-coated QDs (BSA-triP.QDs were analyzed. Then the BSA-coated QDs were conjugated with trastuzumab, which can be recognized and bound specifically to HER2. SKBR-3 breast cancer cells, with over-expression of HER-2, were labeled with hydrophilic QDstrastuzumab and imaged on a fluorescence planar imaging system. Results  The experimental results revealed that BSA as the emulsifying agent, combined with other polymers, can effectively phase transfer hydrophobic QDs. The BSA functionalized QDs exhibited excellent colloidal stability with fine hydrodynamic size distribution (about 70nm in a wide range of pH and ionic strengths values. Moreover, no significant cytotoxic effect was observed in SKBR-3 cells treated with BSA-coated QDs. After being coupled with trastuzumab, the hydrophilic QDs can be used as an immunofluorescence probe for HER2-positive breast cancer cell imaging. Conclusions  The advantages of BSA-coated QDs include straight forward synthesis, high colloidal stability, and promising immunofluorescence characteristic when coupled with specific antibody. These are therefore proved to be ideal nano systems for biomedical labeling, targeting, and imaging. DOI: 10.11855/j.issn.0577-7402.2016.05.04

  4. The topology of plasminogen binding and activation on the surface of human breast cancer cells

    OpenAIRE

    Andronicos, N M; Ranson, M.

    2001-01-01

    The urokinase-dependent activation of plasminogen by breast cancer cells plays an important role in metastasis. We have previously shown that the metastatic breast cancer cell line MDA-MB-231 over-expresses urokinase and binds and efficiently activates plasminogen at the cell surface compared to non-metastatic cells. The aim of this study was to further characterise plasminogen binding and determine the topology of cell surface-bound plasminogen in terms of its potential for activation. The l...

  5. High efficiency solar cells combining a perovskite and a silicon heterojunction solar cells via an optical splitting system

    International Nuclear Information System (INIS)

    We have applied an optical splitting system in order to achieve very high conversion efficiency for a full spectrum multi-junction solar cell. This system consists of multiple solar cells with different band gap optically coupled via an “optical splitter.” An optical splitter is a multi-layered beam splitter with very high reflection in the shorter-wave-length range and very high transmission in the longer-wave-length range. By splitting the incident solar spectrum and distributing it to each solar cell, the solar energy can be managed more efficiently. We have fabricated optical splitters and used them with a wide-gap amorphous silicon (a-Si) solar cell or a CH3NH3PbI3 perovskite solar cell as top cells, combined with mono-crystalline silicon heterojunction (HJ) solar cells as bottom cells. We have achieved with a 550 nm cutoff splitter an active area conversion efficiency of over 25% using a-Si and HJ solar cells and 28% using perovskite and HJ solar cells

  6. High efficiency solar cells combining a perovskite and a silicon heterojunction solar cells via an optical splitting system

    Energy Technology Data Exchange (ETDEWEB)

    Uzu, Hisashi, E-mail: Hisashi.Uzu@kaneka.co.jp, E-mail: npark@skku.edu; Ichikawa, Mitsuru; Hino, Masashi; Nakano, Kunihiro; Meguro, Tomomi; Yamamoto, Kenji [Kaneka Corporation, 5-1-1, Torikai-Nishi, Settsu, Osaka 566-0072 (Japan); Hernández, José Luis [Kaneka Belgium N.V., Nijverheidsstraat 16, 2260 Westerlo-Oevel (Belgium); Kim, Hui-Seon; Park, Nam-Gyu, E-mail: Hisashi.Uzu@kaneka.co.jp, E-mail: npark@skku.edu [School of Chemical Engineering and Department of Energy Science, Sungkyunkwan University, 300 Cheoncheon-dong, Jangan-gu, Suwon 440-746 (Korea, Republic of)

    2015-01-05

    We have applied an optical splitting system in order to achieve very high conversion efficiency for a full spectrum multi-junction solar cell. This system consists of multiple solar cells with different band gap optically coupled via an “optical splitter.” An optical splitter is a multi-layered beam splitter with very high reflection in the shorter-wave-length range and very high transmission in the longer-wave-length range. By splitting the incident solar spectrum and distributing it to each solar cell, the solar energy can be managed more efficiently. We have fabricated optical splitters and used them with a wide-gap amorphous silicon (a-Si) solar cell or a CH{sub 3}NH{sub 3}PbI{sub 3} perovskite solar cell as top cells, combined with mono-crystalline silicon heterojunction (HJ) solar cells as bottom cells. We have achieved with a 550 nm cutoff splitter an active area conversion efficiency of over 25% using a-Si and HJ solar cells and 28% using perovskite and HJ solar cells.

  7. The viability and intestinal epithelial cell adhesion of probiotic strain combination--in vitro study.

    Science.gov (United States)

    Piątek, Jacek; Gibas-Dorna, Magdalena; Olejnik, Anna; Krauss, Hanna; Wierzbicki, Krzysztof; Żukiewicz-Sobczak, Wioletta; Głowacki, Maciej

    2012-01-01

    To be effective, probiotic bacteria must exhibit a number of functional characteristics, including the resistance to gastric acidity and the ability to adhere to the intestinal epithelium. In this study, we examined in vitro the viability of lactic acid bacteria (LAB) combination after exposure to low pH, and the adhesion of LAB to Caco-2 cells during coincubation of 9 bacterial strains. To test bacterial viability, 6 commercially available products were incubated in 0.1 N HCl at pH 1.2 for 60 min. The greatest growth inhibition was noted for the non-capsulated product containing the Lactobacillus rhamnosus strain (log reduction of CFU = 6.4), and the best survival observed for the product containing 9 bacterial strains, equipped with a modern capsule made according to the Multi-Resistant Encapsulation technology (log reduction of CFU = 0.1). In the adhesion experiment, the combination of 9 bacterial strains was added to 17-day-old Caco-2 cell culture for 90 min. The greatest efficiency of adhesion was observed for the inoculum containing 5.5x10(8) CFU/mL/9.6 cm(2) of Caco-2 and the dose of probiotic bacteria of 190 cells per one Caco-2 cell. As a result, approximately 157 bacterial cells adhered to one Caco-2 cell. The results indicate that the combination of 9 bacterial strains in the examined product is characterized as highly adhesive. PMID:22462453

  8. Increasing binding density of yeast cells by control of surface charge with allylamine grafting to ion modified polymer surfaces.

    Science.gov (United States)

    Tran, Clara T H; Kondyurin, Alexey; Chrzanowski, Wojciech; Bilek, Marcela M M; McKenzie, David R

    2014-10-01

    Plasma immersion ion implantation (PIII) treatment of polymers creates a biointerface capable of direct covalent immobilization of biomolecules. The immobilization of protein molecules is achieved by covalent bonds formed between embedded radicals on the treated surface and amino acid side chains and cells can be immobilized through cell-wall proteins. The attachment density of negatively charged entities on a PIII treated surface is inhibited by its negative surface charge at neutral pH. To reduce the negative charge of PIII treated surfaces in phosphate buffer (pH 7.4, 11mM), we develop an effective approach of grafting allylamine monomers onto the treated surface. The results reveal reactions between allylamine and radicals on the PIII treated surface. One of these triggers polymerization, increasing the number of amine groups grafted. As a consequence, the PIII treated polystyrene surface after allylamine exposure becomes more hydrophobic and less negatively charged in phosphate buffer. Using yeast cells as an example, we have shown a significant improvement (6-15 times) of cell density immobilized on the PIII treated surface after exposure to allylamine. PMID:25092587

  9. Growth inhibitory effects of Phyllanthus niruri extracts in combination with cisplatin on cancer cell lines

    Institute of Scientific and Technical Information of China (English)

    Raimundo Fernandes de Araújo Júnior; Luiz Alberto Lira Soares; Cínthia Raquel da Costa Porto; Ranniere Gurgel Furtado de Aquino; Hugo Gon(c)alo Guedes; Pedro Ros Petrovick; Tatiane Pereira de Souza

    2012-01-01

    AIM:To investigate the cytotoxic effects of spray-dried extracts of Phyllanthus niruri in combination with cisplatin on two cancer cell lines.METHODS:Colorectal carcinoma (HT29) and human hepatocellular carcinoma (HepG2) cells were treated with spray-dried extracts of Phy//anthus niruri (SDEPN) either alone or in combination with cisplatin at different concentrations (0.5 mg/mL and 1 mg/mL) for 4 h and 24 h.To verify and quantify cancer cells treated with these products as well as identify the cell cycle stage and cell viability,we stained the cells with propidium iodide and assessed them by flow cytometry.The percentage of cells in different cell cycle phases was quantified and data were expressed as histograms.Significant differences between groups were determined using analysis of variance and Bonferroni's test,as indicated.A value of P < 0.05 was considered to be statistically significant.RESULTS:SDEPN had significantly different cytotoxic effects on HT29 (2.81 ± 0.11 vs 3.51 ± 1.13,P > 0.05) and HepG2 (5.07 ± 0.3 vs 15.9 ± 1.04,P <0.001) cells when compared to control cells for 4 h.SDEPN also had significantly different cytotoxic effects on HT29 (1.91 ± 0.57 vs 4.53 ± 1.22,P > 0.05) and HepG2 (14.56 ± 1.6 vs 35.67 ± 3.94,P < 0.001) cells when compared to control cells for 24 h.Both cell lines were killed by cisplatin in a dose-dependent manner compared to control cells (HepG2 cells for 4 h:10.78 ± 1.58 vs 53.89 ± 1.53,P < 0.001; 24 h:8.9 ± 1.43 vs 62.78 ± 1.87,P < 0.001 and HT29 cells for 4 h:9.52 ± 0.913 vs 49.86 ± 2.89,P < 0.001; 24 h:11.78 ± 1.05 vs 53.34 ± 2.65,P < 0.001).In HT29 cells,pretreatment with SDEPN and subsequent treatment with cisplatin resulted in a greater number of cells being killed (12.78 ± 1.01 vs 93.76 ± 1.6,P < 0.001).HepG2 cells showed significant cell killing with treatment with SDEPN when combined with cisplatin (12.87±2.78 vs 78.8 ± 3.02,P < 0.001).CONCLUSION:SDEPN is selectively toxic

  10. Electrospun fibrous scaffolds combined with nanoscale hydroxyapatite induce osteogenic differentiation of human periodontal ligament cells

    Directory of Open Access Journals (Sweden)

    Wu XN

    2014-08-01

    Full Text Available Xiaonan Wu,1 Leiying Miao,2,# Yingfang Yao,3 Wenlei Wu,1 Yu Liu,1 Xiaofeng Chen,1 Weibin Sun1,# 1Department of Periodontology, Hospital of Stomatology, Medical School of Nanjing University, Nanjing, People’s Republic of China; 2Department of Cariology and Endodontics, Hospital of Stomatology, Medical School of Nanjing University, Nanjing, People’s Republic of China; 3Eco-materials and Renewable Energy Research Center, Department of Materials Science and Engineering, National Laboratory of Solid State Microstructures, Nanjing University, Nanjing, People’s Republic of China #These authors contributed equally to this work Abstract: Periodontal repair is a complex process in which regeneration of alveolar bone is a vital component. The aim of this study was to develop a biodegradable scaffold with good biocompatibility and osteoinductive ability. Two types of composite fibrous scaffolds were produced by electrospinning, ie, type I collagen/poly(є-caprolactone (COL/PCL and type I collagen/poly(є-caprolactone/nanoscale hydroxyapatite (COL/PCL/nHA with an average fiber diameter of about 377 nm. After a simulated body fluid (SBF immersion test, the COL/PCL/nHA-SBF scaffold developed a rough surface because of the calcium phosphate deposited on the fibers, suggesting that the presence of nHA promoted the mineralization potential of the scaffold. Energy dispersive X-ray spectroscopy clearly showed the calcium and phosphorus content in the COL/PCL/nHA and COL/PCL/nHA-SBF scaffolds, confirming the findings of nHA and calcium phosphate precipitation on scanning electron micrographs. Water contact analysis revealed that nHA could improve the hydrophilic nature of the COL/PCL/nHA-SBF scaffold. The morphology of periodontal ligament cells cultured on COL/PCL-SBF and COL/PCL/nHA-SBF was evaluated by scanning electron microscopy. The results showed that cells adhered to either type of scaffold and were slightly spindle-shaped in the beginning, then

  11. Inferring three-dimensional surface displacement field by combining SAR interferometric phase and amplitude information of ascending and descending orbits

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    Conventional Interferometric Synthetic Aperture Radar(InSAR) technology can only measure one-dimensional surface displacement(along the radar line-of-sight(LOS) direction).Here we presents a method to infer three-dimensional surface displacement field by combining SAR interferometric phase and amplitude information of ascending and descending orbits.The method is realized in three steps:(1) measuring surface displacements along the LOS directions of both ascending and descending orbits based on interferometric phases;(2) measuring surface displacements along the azimuth directions of both the ascending and descending orbits based on the SAR amplitude data;and(3) estimating the three-dimensional(3D) surface displacement field by combining the above four independent one-dimensional displacements using the method of least squares and Helmert variance component estimation.We apply the method to infer the 3D surface displacement field caused by the 2003 Bam,Iran,earthquake.The results reveal that in the northern part of Bam the ground surface experienced both subsidence and southwestward horizontal movement,while in the southern part uplift and southeastward horizontal movement occurred.The displacement field thus determined matches the location of the fault very well with the maximal displacements reaching 22,40,and 30 cm,respectively in the up,northing and easting directions.Finally,we compare the 3D displacement field with that simulated from the Okada model.The results demonstrate that the method presented here can be used to generate reliable and highly accurate 3D surface displacement fields.

  12. Celecoxib in combination with retinoid CD437 inhibits melanoma A375 cell in vitro

    Institute of Scientific and Technical Information of China (English)

    Jianwen REN; Zhenhui PENG; Birong GUO; Min PAN

    2009-01-01

    This study aimed to investigate the effects of celecoxib, synthetic retinoid 6-[3-(1-adamantyl)-4-hydroxyphenyl]-2-naphthalenecarboxylicacid (CD437)and the combination of the two on cell proliferation, apoptosis, and cycle arrest of human malignant mela-noma A375 cells. 3-(4,5-dimethylthiazol-2-yl)-2,5-di-phenyltetrazoliumbromide assay (MTT assay) was applied to determine the anti-proliferative effects of the drugs on human malignant melanoma A375 cells. Flow cytometry was performed to investigate the influence of the drugs on cell cycle and cell apoptosis. Both celecoxib and CD437 could inhibit the growth of human malignant melanoma A375 cells in a dose-dependent manner. Celecoxib at 80 μmol/L inhibited proliferation, induced apoptosis and G2/M cell cycle arrest of human malignant melanoma A375 cells after treatment for 24 h [proliferation inhibiting rate: (50.2±2.51)%, apoptosis rate: (35.91±1.80)%]. CD437 at 10μmol/L inhibited proliferation, induced apoptosis and G0/G1 cell cycle arrest of human malignant melanoma A375 cells after treatment for 24 h [proliferation inhibiting rate: (58.6±2.38)%, apoptosis rate: (28.03± 0.77)%]. Celecoxib in combination with CD437 could significantly enhance the effects of inhibiting proliferation and inducing apoptosis of human malignant melanoma A375 cells 24 h after treatment compared with the drugalone [proliferation inhibiting rate: (68.92±1.72)%, apop-tosis rate: (42.09±1.05)%, both P <0.05] and decrease the proportion of the S phase in the cell cycle. Celecoxib could inhibit the growth of human malignant melanoma A375 cells by inducing apoptosis and G2/M cycle arrest. CD437 could inhibit the growth of human malignant melanoma A375 cells by inducing apoptosis and G0/G1 cycle arrest. Celecoxib exhibited additive effects with CD437 on retarding the growth and inducing apoptosis of human malignant melanoma A375 cells. Celecoxib in combination with CD437 may become an effective method for prevention and treatment of

  13. Cell surface molecules and fibronectin-mediated cell adhesion: effect of proteolytic digestion of membrane proteins

    OpenAIRE

    1982-01-01

    Proteases have been used as a tool to investigate the role of surface molecules in fibronectin-mediated cell adhesion. Proteolytic digestion of membrane-proteins by pronase (1 mg/ml for 20 min at 37 degrees C) completely inhibited adhesion of baby hamster kidney (BHK) fibroblasts on fibronectin-coated plastic dishes. Various degrees of inhibition were also obtained after treatment with proteinase K, chymotrypsin, papain, subtilopeptidase A, and thermolysin. Protein synthesis was required to r...

  14. Quantitative comparison of a human cancer cell surface proteome between interphase and mitosis.

    Science.gov (United States)

    Özlü, Nurhan; Qureshi, Mohammad H; Toyoda, Yusuke; Renard, Bernhard Y; Mollaoglu, Gürkan; Özkan, Nazlı E; Bulbul, Selda; Poser, Ina; Timm, Wiebke; Hyman, Anthony A; Mitchison, Timothy J; Steen, Judith A

    2015-01-13

    The cell surface is the cellular compartment responsible for communication with the environment. The interior of mammalian cells undergoes dramatic reorganization when cells enter mitosis. These changes are triggered by activation of the CDK1 kinase and have been studied extensively. In contrast, very little is known of the cell surface changes during cell division. We undertook a quantitative proteomic comparison of cell surface-exposed proteins in human cancer cells that were tightly synchronized in mitosis or interphase. Six hundred and twenty-eight surface and surface-associated proteins in HeLa cells were identified; of these, 27 were significantly enriched at the cell surface in mitosis and 37 in interphase. Using imaging techniques, we confirmed the mitosis-selective cell surface localization of protocadherin PCDH7, a member of a family with anti-adhesive roles in embryos. We show that PCDH7 is required for development of full mitotic rounding pressure at the onset of mitosis. Our analysis provided basic information on how cell cycle progression affects the cell surface. It also provides potential pharmacodynamic biomarkers for anti-mitotic cancer chemotherapy.

  15. Coupling Binding to Catalysis – Using Yeast Cell Surface Display to Select Enzymatic Activities

    OpenAIRE

    Zhang, Keya; Bhuripanyo, Karan; Wang, Yiyang; Yin, Jun

    2015-01-01

    We find yeast cell surface display can be used to engineer enzymes by selecting the enzyme library for high affinity binding to reaction intermediates. Here we cover key steps of enzyme engineering on the yeast cell surface including library design, construction, and selection based on magnetic and fluorescence activated cell sorting.

  16. Coupling Binding to Catalysis: Using Yeast Cell Surface Display to Select Enzymatic Activities.

    Science.gov (United States)

    Zhang, Keya; Bhuripanyo, Karan; Wang, Yiyang; Yin, Jun

    2015-01-01

    We find yeast cell surface display can be used to engineer enzymes by selecting the enzyme library for high affinity binding to reaction intermediates. Here we cover key steps of enzyme engineering on the yeast cell surface including library design, construction, and selection based on magnetic and fluorescence-activated cell sorting. PMID:26060080

  17. Distribution, Arrangement and Interconnectedness of Cell Surface Receptor sites in the body of an Organism

    Directory of Open Access Journals (Sweden)

    Utoh-Nedosa

    2011-01-01

    Full Text Available Cell surface receptors have been identified as the sites of disease infectivity in living organisms in a previous study. Drugs used for the treatment or cure of infections have to eliminate infections through attacking infective organisms at the cell surface receptors to which the infective organisms are attached. Problem statement: The present study examines a wide sample of living things to get more information on the relationship of one cell surface receptor to other cell surface receptors in the body of an organism. Approach: The arrangement of cell surface receptors on the external covering of a few samples of fruits, leaves, stems, dry wood of a plant; wall gecko and some parts of the human body, were examined and photographed. Transverse and/or Longitudinal sections of soursop fruit and sycamore fruit were also examined and photographed. The five different coverings of the fleshy part of a coconut were also photographed. The photographs were studied to note the relationship of disease infection attached to cell surface receptors on the external surface of an organ to disease infection on the innermost covering of the same organ. Results: The results of the study showed that all living things had ubiquitous distribution of cell surface receptors which are usually observable with the unaided eye as dots or spots on the external covering of an organ, tissue or cell. The dots or receptor sites of cell surface receptors in the study are arranged in lines which were perpendicular, oblique, transverse or arranged in any other lineal geometrical form. The lineally arranged cell surface receptors were noted to be connected by grooves, channels or pipes which joined other receptor channels or intersected with them. Smaller cell surface receptor channels emptied into bigger channels or continued as small sized channels that ran side by side in a connective tissue bundle. These connective tissue bundles that carried many independent small-sized cell

  18. Combined model of non-conformal layer growth for accurate optical simulation of thin-film silicon solar cells

    Energy Technology Data Exchange (ETDEWEB)

    Sever, M.; Lipovsek, B.; Krc, J.; Campa, A.; Topic, M. [University of Ljubljana, Faculty of Electrical Engineering Trzaska cesta 25, Ljubljana 1000 (Slovenia); Sanchez Plaza, G. [Technical University of Valencia, Valencia Nanophotonics Technology Center (NTC) Valencia 46022 (Spain); Haug, F.J. [Ecole Polytechnique Federale de Lausanne EPFL, Institute of Microengineering IMT, Photovoltaics and Thin-Film Electronics Laboratory, Neuchatel 2000 (Switzerland); Duchamp, M. [Ernst Ruska-Centre for Microscopy and Spectroscopy with Electrons Institute for Microstructure Research, Research Centre Juelich, Juelich D-52425 (Germany); Soppe, W. [ECN-Solliance, High Tech Campus 5, Eindhoven 5656 AE (Netherlands)

    2013-12-15

    In thin-film silicon solar cells textured interfaces are introduced, leading to improved antireflection and light trapping capabilities of the devices. Thin-layers are deposited on surface-textured substrates or superstrates and the texture is translated to internal interfaces. For accurate optical modelling of the thin-film silicon solar cells it is important to define and include the morphology of textured interfaces as realistic as possible. In this paper we present a model of thin-layer growth on textured surfaces which combines two growth principles: conformal and isotropic one. With the model we can predict the morphology of subsequent internal interfaces in thin-film silicon solar cells based on the known morphology of the substrate or superstrate. Calibration of the model for different materials grown under certain conditions is done on various cross-sectional scanning electron microscopy images of realistic devices. Advantages over existing growth modelling approaches are demonstrated - one of them is the ability of the model to predict and omit the textures with high possibility of defective regions formation inside the Si absorber layers. The developed model of layer growth is used in rigorous 3-D optical simulations employing the COMSOL simulator. A sinusoidal texture of the substrate is optimised for the case of a micromorph silicon solar cell. More than a 50 % increase in short-circuit current density of the bottom cell with respect to the flat case is predicted, considering the defect-free absorber layers. The developed approach enables accurate prediction and powerful design of current-matched top and bottom cell.

  19. The advantages of combining ion beam techniques for surface layer analysis

    International Nuclear Information System (INIS)

    The problem of elemental composition analysis for a variable thickness surface layer has been investigated. Both protons and alpha-particles with a range of energy were used as the incident ions for PIXE, PIGME and RBS measurements so that the variation of elemental composition as a function of surface thickness could be studied. An example of measurements on desert varnish coatings on underlying rock is given

  20. The effect of cerium valence states at cerium oxide nanoparticle surfaces on cell proliferation

    KAUST Repository

    Naganuma, Tamaki

    2014-05-01

    Understanding and controlling cell proliferation on biomaterial surfaces is critical for scaffold/artificial-niche design in tissue engineering. The mechanism by which underlying integrin ligates with functionalized biomaterials to induce cell proliferation is still not completely understood. In this study, poly-l-lactide (PL) scaffold surfaces were functionalized using layers of cerium oxide nanoparticles (CNPs), which have recently attracted attention for use in therapeutic application due to their catalytic ability of Ce4+ and Ce3+ sites. To isolate the influence of Ce valance states of CNPs on cell proliferation, human mesenchymal stem cells (hMSCs) and osteoblast-like cells (MG63) were cultured on the PL/CNP surfaces with dominant Ce4+ and Ce3+ regions. Despite cell type (hMSCs and MG63 cells), different surface features of Ce4+ and Ce3+ regions clearly promoted and inhibited cell spreading, migration and adhesion behavior, resulting in rapid and slow cell proliferation, respectively. Cell proliferation results of various modified CNPs with different surface charge and hydrophobicity/hydrophilicity, indicate that Ce valence states closely correlated with the specific cell morphologies and cell-material interactions that trigger cell proliferation. This finding suggests that the cell-material interactions, which influence cell proliferation, may be controlled by introduction of metal elements with different valence states onto the biomaterial surface. © 2014 Elsevier Ltd.

  1. CpG Oligodeoxynucleotide1826 combined with radioresistant cancer cell vaccine confers significant antitumor effects.

    Science.gov (United States)

    Zhuang, X B; Xing, N; Zhang, Q; Yuan, S J; Chen, W; Qiao, T K

    2015-01-01

    Immunotherapy is a hot issue in cancer research over the years and tumor cell vaccine is one of the increasing number of studies. Although the whole tumor cell vaccine can provide the best source of immunizing antigens, there is still a limitation that most tumors are not naturally immunogenic. CpG Oligodeoxynucleotides (CpG ODNs), synthetic oligonucleotides containing a cytosine-phosphate-guanine(CpG) motif, was shown to enhance immune responses to a wide variety of antigens. In this study, we generated the radioresistant Lewis lung cancer cell by repeated X-ray radiation and inactivated it as a whole tumor cell vaccine to enhance the immunogenicity of tumor cell vaccine. Mice were subcutaneously immunized with this inactivated vaccine combined with CpG ODN1826 and then inoculated with autologous Lewis lung cancer (LLC) to estimate the antitumor efficacy. The results showed that the radioresistant tumor cell vaccine combined with CpG ODN1826 could significantly inhibit tumor growth, increased survival of the mice and with 20% of the mice surviving tumor free in vivo compared with the unimmunized mice bearing LLC tumor. A significant increase of apoptosis was also observed in the tumor prophylactically immunized with vaccine of inactivated radioresistant tumor cell plus CpG ODN1826. The potent antitumor effect correlated with higher secretion levels of tumor necrosis factor-alpha(TNF-α) and lower levels of interleukin-10(IL-10) concentration in serum. Furthermore, the results suggested that the antitumor mechanism was probably depended on the decreased level of programmed death ligand-1(PD-L1) which plays an important role in the negative regulation of immune response by the inhibition of tumor antigen-specific T cell activation. These findings clearly demonstrated that the radioresistant tumor cell vaccine combined with CpG ODN1826 as an appropriate adjuvant could induce effective antitumor immunity in vivo. PMID:26458317

  2. Combined effects of EGFR tyrosine kinase inhibitors and vATPase inhibitors in NSCLC cells

    Energy Technology Data Exchange (ETDEWEB)

    Jin, Hyeon-Ok [KIRAMS Radiation Biobank, Korea Institute of Radiological and Medical Sciences, 75 Nowon-ro, Nowon-gu, Seoul, 139–706 (Korea, Republic of); Hong, Sung-Eun [Division of Radiation Cancer Research, Korea Institute of Radiological and Medical Sciences, 75 Nowon-ro, Nowon-gu, Seoul, 139–706 (Korea, Republic of); Kim, Chang Soon [Department of Microbiological Engineering, Kon-Kuk University, 120 Neungdong-ro, Gwangjin-gu, Seoul, 143–701 (Korea, Republic of); Park, Jin-Ah; Kim, Jin-Hee; Kim, Ji-Young; Kim, Bora [KIRAMS Radiation Biobank, Korea Institute of Radiological and Medical Sciences, 75 Nowon-ro, Nowon-gu, Seoul, 139–706 (Korea, Republic of); Chang, Yoon Hwan; Hong, Seok-Il; Hong, Young Jun [Department of Laboratory Medicine, Korea Cancer Center Hospital, Korea Institute of Radiological and Medical Sciences, 75 Nowon-ro, Nowon-gu, Seoul, 139–706 (Korea, Republic of); Park, In-Chul, E-mail: parkic@kirams.re.kr [Division of Radiation Cancer Research, Korea Institute of Radiological and Medical Sciences, 75 Nowon-ro, Nowon-gu, Seoul, 139–706 (Korea, Republic of); Lee, Jin Kyung, E-mail: jklee@kirams.re.kr [KIRAMS Radiation Biobank, Korea Institute of Radiological and Medical Sciences, 75 Nowon-ro, Nowon-gu, Seoul, 139–706 (Korea, Republic of); Department of Laboratory Medicine, Korea Cancer Center Hospital, Korea Institute of Radiological and Medical Sciences, 75 Nowon-ro, Nowon-gu, Seoul, 139–706 (Korea, Republic of)

    2015-08-15

    Despite excellent initial clinical responses of non-small cell lung cancer (NSCLC) patients to epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKIs), many patients eventually develop resistance. According to a recent report, vacuolar H + ATPase (vATPase) is overexpressed and is associated with chemotherapy drug resistance in NSCLC. We investigated the combined effects of EGFR TKIs and vATPase inhibitors and their underlying mechanisms in the regulation of NSCLC cell death. We found that combined treatment with EGFR TKIs (erlotinib, gefitinib, or lapatinib) and vATPase inhibitors (bafilomycin A1 or concanamycin A) enhanced synergistic cell death compared to treatments with each drug alone. Treatment with bafilomycin A1 or concanamycin A led to the induction of Bnip3 expression in an Hif-1α dependent manner. Knock-down of Hif-1α or Bnip3 by siRNA further enhanced cell death induced by bafilomycin A1, suggesting that Hif-1α/Bnip3 induction promoted resistance to cell death induced by the vATPase inhibitors. EGFR TKIs suppressed Hif-1α and Bnip3 expression induced by the vATPase inhibitors, suggesting that they enhanced the sensitivity of the cells to these inhibitors by decreasing Hif-1α/Bnip3 expression. Taken together, we conclude that EGFR TKIs enhance the sensitivity of NSCLC cells to vATPase inhibitors by decreasing Hif-1α/Bnip3 expression. We suggest that combined treatment with EGFR TKIs and vATPase inhibitors is potentially effective for the treatment of NSCLC. - Highlights: • Co-treatment with EGFR TKIs and vATPase inhibitors induces synergistic cell death • EGFR TKIs enhance cell sensitivity to vATPase inhibitors via Hif-1α downregulation • Co-treatment of these inhibitors is potentially effective for the treatment of NSCLC.

  3. Structured Light Based 3d Scanning for Specular Surface by the Combination of Gray Code and Phase Shifting

    Science.gov (United States)

    Zhang, Yujia; Yilmaz, Alper

    2016-06-01

    Surface reconstruction using coded structured light is considered one of the most reliable techniques for high-quality 3D scanning. With a calibrated projector-camera stereo system, a light pattern is projected onto the scene and imaged by the camera. Correspondences between projected and recovered patterns are computed in the decoding process, which is used to generate 3D point cloud of the surface. However, the indirect illumination effects on the surface, such as subsurface scattering and interreflections, will raise the difficulties in reconstruction. In this paper, we apply maximum min-SW gray code to reduce the indirect illumination effects of the specular surface. We also analysis the errors when comparing the maximum min-SW gray code and the conventional gray code, which justifies that the maximum min-SW gray code has significant superiority to reduce the indirect illumination effects. To achieve sub-pixel accuracy, we project high frequency sinusoidal patterns onto the scene simultaneously. But for specular surface, the high frequency patterns are susceptible to decoding errors. Incorrect decoding of high frequency patterns will result in a loss of depth resolution. Our method to resolve this problem is combining the low frequency maximum min-SW gray code and the high frequency phase shifting code, which achieves dense 3D reconstruction for specular surface. Our contributions include: (i) A complete setup of the structured light based 3D scanning system; (ii) A novel combination technique of the maximum min-SW gray code and phase shifting code. First, phase shifting decoding with sub-pixel accuracy. Then, the maximum min-SW gray code is used to resolve the ambiguity resolution. According to the experimental results and data analysis, our structured light based 3D scanning system enables high quality dense reconstruction of scenes with a small number of images. Qualitative and quantitative comparisons are performed to extract the advantages of our new

  4. Cell surface estrogen receptor alpha is upregulated during subchronic metabolic stress and inhibits neuronal cell degeneration.

    Directory of Open Access Journals (Sweden)

    Cristiana Barbati

    Full Text Available In addition to the classical nuclear estrogen receptor, the expression of non-nuclear estrogen receptors localized to the cell surface membrane (mER has recently been demonstrated. Estrogen and its receptors have been implicated in the development or progression of numerous neurodegenerative disorders. Furthermore, the pathogenesis of these diseases has been associated with disturbances of two key cellular programs: apoptosis and autophagy. An excess of apoptosis or a defect in autophagy has been implicated in neurodegeneration. The aim of this study was to clarify the role of ER in determining neuronal cell fate and the possible implication of these receptors in regulating either apoptosis or autophagy. The human neuronal cell line SH-SY5Y and mouse neuronal cells in primary culture were thus exposed to chronic minimal peroxide treatment (CMP, a form of subcytotoxic minimal chronic stress previously that mimics multiple aspects of long-term cell stress and represents a limited molecular proxy for neurodegenerative processes. We actually found that either E2 or E2-bovine serum albumin construct (E2BSA, i.e. a non-permeant form of E2 was capable of modulating intracellular cell signals and regulating cell survival and death. In particular, under CMP, the up-regulation of mERα, but not mERβ, was associated with functional signals (ERK phosphorylation and p38 dephosphorylation compatible with autophagic cytoprotection triggering and leading to cell survival. The mERα trafficking appeared to be independent of the microfilament system cytoskeletal network but was seemingly associated with microtubular apparatus network, i.e., to MAP2 molecular chaperone. Importantly, antioxidant treatments, administration of siRNA to ERα, or the presence of antagonist of ERα hindered these events. These results support that the surface expression of mERα plays a pivotal role in determining cell fate, and that ligand-induced activation of mER signalling exerts a

  5. Chronic exposure to combined carcinogens enhances breast cell carcinogenesis with mesenchymal and stem-like cell properties.

    Directory of Open Access Journals (Sweden)

    Lenora Ann Pluchino

    Full Text Available Breast cancer is the most common type of cancer affecting women in North America and Europe. More than 85% of breast cancers are sporadic and attributable to long-term exposure to small quantities of multiple carcinogens. To understand how multiple carcinogens act together to induce cellular carcinogenesis, we studied the activity of environmental carcinogens 4-(methylnitrosamino-1-(3-pyridyl-1-butanone (NNK and benzo[a]pyrene (B[a]P, and dietary carcinogen 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP using our breast cell carcinogenesis model. Our study revealed, for the first time, that combined NNK and B[a]P enhanced breast cell carcinogenesis chronically induced by PhIP in both non-cancerous and cancerous breast cells. Co-exposure was more potent than sequential exposure to combined NNK and B[a]P followed by PhIP in inducing carcinogenesis. Initiation of carcinogenesis was measured by transient endpoints induced in a single exposure, while progression of carcinogenesis was measured by acquisition of constitutive endpoints in cumulative exposures. Transient endpoints included DNA damage, Ras-Erk-Nox pathway activation, reactive oxygen species elevation, and increased cellular proliferation. Constitutive endpoints included various cancer-associated properties and signaling modulators, as well as enrichment of cancer stem-like cell population and activation of the epithelial-to-mesenchymal transition program. Using transient and constitutive endpoints as targets, we detected that a combination of the green tea catechins ECG and EGCG, at non-cytotoxic levels, was more effective than individual agents in intervention of cellular carcinogenesis induced by combined NNK, B[a]P, and PhIP. Thus, use of combined ECG and EGCG should be seriously considered for early intervention of breast cell carcinogenesis associated with long-term exposure to environmental and dietary carcinogens.

  6. Effect of combined electromagnetic fields on surface quality and structure of a HDC casting 5182 Al alloy ingot

    Institute of Scientific and Technical Information of China (English)

    Qingfeng ZHU; Zhihao ZHAO; Yubo ZUO; Xiangjie WANG; Jianzhong CUI

    2012-01-01

    5182 Al alloy ingots with the diameters of 100 mm are produced by conventional and combined electromagnetic fields (CEMF) horizontal direction chill (HDC) casting process.The effect of CEMF on the ingot surface quality and floating grains is investigated.The results show that the CEMF is effective in reducing the segregation layer in both top and bottom sub-surfaces.Big floating grains with coarse dendrite arms mainly distribute in the crescent-shaped area near the bottom in the conventional HDC casting ingot.With the application of the CEMF,they disappear in the whole cross section of the HDC casting ingot and the equiaxed grains are significantly refined.

  7. Treatment of B-cells non-Hodgkin lymphomas with combined immunochemotherapy: ability to treatment optimization

    OpenAIRE

    Smirnova, N. V.; N. V. Myakova; M. B. Belogurova; O. V. Ryskal; O. E. Nikonova; G. R. Sharapova; A. S. Fedorova; N. A. Grigorieva; A. V. Shamardina; N. I. Ponomareva; D. S. Abramov; D. M. Konovalov; M. E. Dubrovina; A. A. Maschan; E. V. Samochatova

    2015-01-01

    The results of two consecutive multicenter clinical trials enrolled 241 patient with childhood mature B-cells non-Hodgkin lymphomas/leukemia are presented. Patients received treatment according B-NHL 2004mab protocol (n = 83) and B-NHL 2010M (n = 158) with combined immunochemotherapy (ICT) in Russian and Belarus pediatric clinics from 2004 to 2015 years. Primary patients with different mature B-NHL (Burkitt lymphoma/leukemia, diffuse large B-cell lymphoma and primary mediastinal B-cell lympho...

  8. Construction and characterization of spherical Si solar cells combined with SiC electric power inverter

    Science.gov (United States)

    Oku, Takeo; Matsumoto, Taisuke; Hiramatsu, Kouichi; Yasuda, Masashi; Shimono, Akio; Takeda, Yoshikazu; Murozono, Mikio

    2015-02-01

    Spherical silicon (Si) photovoltaic solar cell systems combined with an electric power inverter using silicon carbide (SiC) field-effect transistor (FET) were constructed and characterized, which were compared with an ordinary Si-based converter. The SiC-FET devices were introduced in the direct current-alternating current (DC-AC) converter, which was connected with the solar panels. The spherical Si solar cells were used as the power sources, and the spherical Si panels are lighter and more flexible compared with the ordinary flat Si solar panels. Conversion efficiencies of the spherical Si solar cells were improved by using the SiC-FET.

  9. Combination of Small Molecule Microarray and Confocal Microscopy Techniques for Live Cell Staining Fluorescent Dye Discovery

    Directory of Open Access Journals (Sweden)

    Attila Bokros

    2013-08-01

    Full Text Available Discovering new fluorochromes is significantly advanced by high-throughput screening (HTS methods. In the present study a combination of small molecule microarray (SMM prescreening and confocal laser scanning microscopy (CLSM was developed in order to discover novel cell staining fluorescent dyes. Compounds with high native fluorescence were selected from a 14,585-member library and further tested on living cells under the microscope. Eleven compartment-specific, cell-permeable (or plasma membrane-targeted fluorochromes were identified. Their cytotoxicity was tested and found that between 1–10 micromolar range, they were non-toxic even during long-term incubations.

  10. Combined Effects of Nonylphenol and Bisphenol A on the Human Prostate Epithelial Cell Line RWPE-1

    Directory of Open Access Journals (Sweden)

    Weidong Gan

    2015-04-01

    Full Text Available The xenoestrogens nonylphenol (NP and bisphenol A (BPA are regarded as endocrine disrupting chemicals (EDCs which have widespread occurrence in our daily life. In the present study, the purpose was to analyze the combined effects of NP and BPA on the human prostate epithelial cell line RWPE-1 using two mathematical models based on the Loewe additivity (LA theory and the Bliss independence (BI theory. RWPE-1 cells were treated with NP (0.01–100 µM and BPA (1–5000 µM in either a single or a combined format. A cell viability assay and lactate dehydrogenase (LDH leakage rate assay were employed as endpoints. As predicted by the two models and based on the cell