247 results for qubit oscillator frequency
Integrated miRNA and mRNA profiling of tumor-educated macrophages identifies prognostic subgroups in estrogen receptor-positive breast cancer
Contributors: Annalen Bleckmann, Andreas Leha, Stephan Artmann, Kerstin Menck, Gabriela Salinas-Riester, Claudia Binder, Tobias Pukrop, Tim Beissbarth, Florian Klemm
Stable feature selection of miR-iTEM in breast cancer datasets. List of stably selected miRNAs from the different miRNA sets (miR-iTEM list, miR-Oxford ER + list, all miRNAs list) and their corresponding inclusion frequencies (in % of all cross validation runs) for the two external breast cancer datasets. “-“ indicates that this miRNA was not present in the respective list of miRNAs. ...Stable feature selection of miR-iTEM in breast cancer datasets. List of stably selected miRNAs from the different miRNA sets (miR-iTEM list, miR-Oxford ER + list, all miRNAs list) and their corresponding inclusion frequencies (in % of all cross validation runs) for the two external breast cancer datasets. “-“ indicates that this miRNA was not present in the respective list of miRNAs. ... Various studies have identified aberrantly expressed miRNAs in breast cancer and demonstrated an association between distinct miRNAs and malignant progression as well as metastasis. Even though tumor-associated macrophages (TAM) are known mediators of these processes, little is known regarding their miRNA expression upon education by malignant cells in vivo.
Changes of defense proteins in the extracellular proteome of grapevine (Vitis vinifera cv. Gamay) cell cultures in response to elicitors
Contributors: M.J. Martinez-Esteso, S. Sellés-Marchart, J.C. Vera-Urbina, M.A. Pedreño, R. Bru-Martinez
... In plant cells, elicitors induce defense responses that resemble those triggered by pathogen attack, such as the synthesis of phytoalexins and pathogen-related proteins which accumulate in the extracellular space. In the search for the particular proteins involved in defense responses, we investigated the changes in the extracellular proteome of a grapevine (Vitis vinifera cv. Gamay) cell suspension in response to elicitation with methylated cyclodextrins (MBCD) and methyl jasmonate (MeJA). Twenty-five of the 39 spots differentially expressed in 2-D gels were identified and found to be encoded by 10 different genes: three secretory peroxidases, chitinase-III, β-1,3-glucanase, thaumatin-like, SGNH plant lipase-like, NtPR27-like, xyloglucan endotransglycosylase and subtilisin-like protease. Most of them belong to the pathogenesis-related type proteins. A new class III secretory basic peroxidase and chitinase III were strongly induced in cultures treated with MBCD alone or combined with MeJA, while cultures treated with MeJA alone displayed a general repression of most of the extracellular proteins. Some of the proteins induced in grapevine cell cultures by MBCD are induced in other species by activators of systemic acquired resistance (SAR), a form of plant immunity. Collectively, the results suggest that treatment with MBCD resembles the effect of SAR induction agents in cell cultures.
Brief communication - Expression patterns of photoperiod and temperature regulated heading date genes in Oryza sativa
Contributors: Mallikarjuna Rao Kovi, Gaurav Sablok, XuFeng Bai, Micael Wendell, Odd-Arne Rognli, HuiHui Yu, YongZhong Xing
... In plants, flowering is a major biological phenomenon, which is regulated by an array of interactions occurring between biotic and abiotic factors. In our study, we have compared the expression profiles of flowering genes involved in the flowering pathway, which are influenced by conditions like photoperiod and temperature from seedling to heading developmental stages in two Oryza sativa indica varieties, viz., Zhenshan 97 and Minghui 63 using a expression network approach. Using the network expression approach, we found 17 co-expressed genes having the same expression profile pattern as three key photoperiod flowering genes Hd1, Ehd1 and Hd3a. We also demonstrated that these three co-expressed genes have a similar simulation pattern as temperature flowering genes. Based on our observations, we hypothesize that photoperiod and temperature regulate flowering pathways independently. The present study provides a basis for understanding the network of co-expressed genes involved in flowering pathway and presents a way to demonstrate the behavior of specific gene sets in specific cultivars.
The regulation of focal adhesion complex formation and salivary gland epithelial cell organization by nanofibrous PLGA scaffolds
Contributors: Sharon J. Sequeira, David A. Soscia, Basak Oztan, Aaron P. Mosier, Riffard Jean-Gilles, Anand Gadre, Nathaniel C. Cady, Bülent Yener, James Castracane, Melinda Larsen
... Nanofiber scaffolds have been useful for engineering tissues derived from mesenchymal cells, but few studies have investigated their applicability for epithelial cell-derived tissues. In this study, we generated nanofiber (250 nm) or microfiber (1200 nm) scaffolds via electrospinning from the polymer, poly-l-lactic-co-glycolic acid (PLGA). Cell-scaffold contacts were visualized using fluorescent immunocytochemistry and laser scanning confocal microscopy. Focal adhesion (FA) proteins, such as phosphorylated FAK (Tyr397), paxillin (Tyr118), talin and vinculin were localized to FA complexes in adult cells grown on planar surfaces but were reduced and diffusely localized in cells grown on nanofiber surfaces, similar to the pattern observed in adult mouse salivary gland tissues. Significant differences in epithelial cell morphology and cell clustering were also observed and quantified, using image segmentation and computational cell-graph analyses. No statistically significant differences in scaffold stiffness between planar PLGA film controls compared to nanofibers scaffolds were detected using nanoindentation with atomic force microscopy, indicating that scaffold topography rather than mechanical properties accounts for changes in cell attachments and cell structure. Finally, PLGA nanofiber scaffolds could support the spontaneous self-organization and branching of dissociated embryonic salivary gland cells. Nanofiber scaffolds may therefore have applicability in the future for engineering an artificial salivary gland.
Contributors: Ke Li, Henrieta Fazekasova, Naiyin Wang, Pervinder Sagoo, Qi Peng, Wafa Khamri, Chantelle Gomes, Steven H. Sacks, Giovanna Lombardi, Wuding Zhou
... Integration of innate and adaptive arms of the immune response at a cellular and molecular level appears to be fundamental to the development of powerful effector functions in host defence and aberrant immune responses. Here we provide evidence that the functions of human complement activation and antigen presentation converge on dendritic cells (DCs). We show that several subsets of human DCs [i.e., monocyte derived (CD1a+CD14−), dermal (CD1a+DC-SIGN+), Langerhans (CD1a+Langerin+), myeloid (CD1c+CD19−), plamacytoid (CD45RA+CD123+)] express many of the components of the classical and alternative and terminal pathways of complement. Moreover human DCs have receptors known to detect the biologically active peptides C3a and C5a (C3aR, C5aR) and the covalently bound fragments C3b and metabolites iC3b and C3d which serve in immune adhesion (i.e., CR3, CR4, CRIg). We also show that the human DC surface is characterised by membrane bound regulators of complement activation, which are also known to participate in intracellular signalling (i.e., CD46, CD55, CD59). This work provides an extensive description of complement components relevant to the integrated actions of complement and DC, illuminated by animal studies. It acts as a resource that allows further understanding and exploitation of role of complement in human health and immune mediated diseases.
The control of neural cell-to-cell interactions through non-contact electrical field stimulation using graphene electrodes
Contributors: Chaejeong Heo, Jeongwan Yoo, Siyoung Lee, Areum Jo, Susie Jung, Hyosun Yoo, Young Hee Lee, Minah Suh
... Electric field stimulation has become one of the most promising therapies for a variety of neurological diseases. However, the safety and effectiveness of the stimulator are critical in determining the outcome. Because there are few safe and effective in vivo and/or in vitro stimulator devices, we demonstrate a method that allows for non-contact electric field stimulation with a specific strength that is able to control cell-to-cell interaction in vitro. Graphene, a form of graphite, and polyethylene terephthalate (PET) was used to create a non-cytotoxic in vitro graphene/PET film stimulator. A transient non-contact electric field was produced by charge-balanced biphasic stimuli through the graphene/PET film electrodes and applied to cultured neural cells. We found that weak electric field stimulation (pulse duration of 10 s) as low as 4.5 mV/mm for 32 min was particularly effective in shaping cell-to-cell interaction. Under weak electric field stimulation, we observed a significant increase in the number of cells forming new cell-to-cell couplings and in the number of cells strengthening existing cell-to-cell couplings. The underlying mechanism of the altered cellular interactions may be related to an altered regulation of the endogenous cytoskeletal proteins fibronectin, actin, and vinculin. In conclusion, this technique may open a new therapeutic approach for augmenting cell-to-cell coupling in cell transplantation therapy in the central nervous system.
Contributors: Sang Myoung Noh, Su Eun Han, Gayong Shim, Kyoung Eun Lee, Chan-Wha Kim, Sung Sik Han, Yongseok Choi, Young Keun Kim, Won-Ki Kim, Yu-Kyoung Oh
... Amphiphilic α-tocopherol oligochitosan conjugates were constructed by conjugating α-tocopherol succinate to water soluble oligochitosans with various molecular weights. In aqueous medium, the tocopherol oligochitosan conjugates self-assembled to single layered oligomersomes. The sizes of α-tocopherol-oligochitosan-based oligomersomes (TCOsomes) could be controlled by chain lengths of oligochitosans. The mean sizes of TCOsomes were 220 and 377 nm as the sizes of oligochitosans were 4000 and 12,500, respectively. For all TCOsomes formed in this study, polydispersity indexes were in the ranges of 0.111–0.256. Cryo-TEM images showed clear thickening in the unilamellar layer of TCOsomes upon complexation with siRNAs. Zeta potentials decreased as the ratios of siRNA/TCOsomes increased. TCOsomes self-assembled from tocopherol-oligochitosan 4K (TCOsome4K) significantly enhanced the cellular uptake of siRNAs (>98%), and reduced the expression of target proteins more effectively than did Lipofectamine 2000. In tumor xenografted mice, the intratumoral administration of siMcl-1 using TCOsomes substantially silenced the expression of Mcl-1 and prevented the growth of tumor. The hematoxylin-eosin staining showed the apoptosis of cells in the tissues of the mice treated with siMcl-1/TCOsome4K complexes, but not with siGL2/TCOsome4K complexes. The self-assembling and size-controllable oligomersomes might be suitable for effective in vivo delivery of siRNAs.
Prostaglandin-D synthetase induces transcription of the LH beta subunit in the primary culture of chicken anterior pituitary cells via the PPAR signaling pathway
Contributors: L.-R. Chen, S.-C. Lee, Y.-P. Lin, Y.-L. Hsieh, Y.-L. Chen, J.-R. Yang, J.-F. Liou, C.-F. Chen, Y.-P. Lee, Y.-L. Shiue
... Downstream effects of prostaglandin-D synthetase (PGDS) in a primary culture of chicken (Gallus gallus) anterior pituitary cells were investigated to study how PGDS regulated laying in hens. Either PGDS downstream metabolite, PGD2 or PGJ2, elevated LHB mRNA and LHB protein levels in dose- and time-dependent manners, and treatment with arachidonic acid (1μM) alone upregulated 15-deoxy-Δ12,14-PGJ2 (15-d-PGJ2; derived from PGJ2)/PGJ2, LHB mRNA, and LHB protein levels (P<0.05) in the primary culture of chicken pituitary anterior cells. Transfection of the plasmid Enhanced Green Fluorescent Protein-PGDS plasmid into these cells in medium containing 1μM arachidonic acid additionally increased 15-d-PGJ2/PGJ2, LHB mRNA, and LHB protein levels (P<0.05). In the hypothalamus/pituitary gland of laying hens, there was a high correlation (r=0.64; P<0.05) between PGDS and the peroxisome proliferator-activated receptor A (PPARA) mRNA level in a high egg production strain, the L2 Taiwan country chickens. In commercial Single-Comb White Leghorn layers, there were high correlation coefficients between PGDS and PPARA (r=0.65; P<0.05) and between PGDS and PPARG (r=0.67; P<0.01) mRNA levels. A broad-range PPARs agonist, GW9578 (5 to 500nM), enhanced LHB mRNA and LHB protein levels (P<0.05). The PPARA-specific (GW6471) and PPARG-specific (T0070907) antagonists suppressed endogenous LHB mRNA and LHB protein levels (P<0.05); in addition, both antagonists attenuated arachidonic acid–induced LHB mRNA levels (P<0.05) and PGDS-induced (in the presence of 1μM arachidonic acid) LHB mRNA and LHB protein (P<0.05) levels in the primary culture of chicken anterior pituitary cells. Higher LHB mRNA/LHB protein ratios in PGD2-, PGJ2-, arachidonic acid-, PGDS-, and GW9578-induced as well as GW6471- and T0070907-suppressed anterior pituitary cells suggested that LHB transcription occurred before translation. In conclusion, PGDS induced LHB transcription and subsequent translation via the PPAR signaling pathway.
Species specialization in cytokine biology: Is interleukin-4 central to the TH1–TH2 paradigm in swine?
Contributors: Michael P. Murtaugh, Craig R. Johnson, Zhengguo Xiao, Ronald W. Scamurra, Yaling Zhou
... The TH1–TH2 paradigm provides an elegant model of directed response to infectious pathogens. Developed in the mouse, the model has provided a framework for systematic and mechanistic studies of immune regulation, protective immunity, and vaccine development in swine. Interleukin-4 (IL-4) plays a central role in the paradigm as a regulatory molecule directing development of the TH2 phenotype, as a developmental cytokine essential for antibody production, and as a soluble diagnostic marker of the TH2 cell type. In contrast, while characterizing the biological properties of porcine IL-4, we discovered that it was not a stimulatory factor for porcine B cells. Rather, it blocked antibody and IL-6 secretion and suppressed antigen-stimulated proliferation of B cells. Inhibition was not reversed by treatment with IL-2 and IL-6 treatment. IL-4 did not stimulate T lymphocyte proliferation, but induced cell growth in lymphoblasts in a dose-dependent fashion. These results suggest that IL-4 plays a different role in pigs than in mice and humans, in which it stimulates B cells and is essential for antibody production. Furthermore, the functions of IL-4 in swine cannot be inferred from results in model systems such as the mouse. General models of disease resistance show substantial variation between pigs and mice at the cellular and molecular level. Advances in somatic cell technologies and animal engineering to enable gene knockouts in pigs, in combination with a continuously expanding immunological toolkit, promise an exciting future for pig immunology, detailed mechanistic elucidation of the TH1–TH2 paradigm, and an improved understanding of the role of IL-4 in porcine immunity to infectious disease.
Contributors: Corinna Sedlak, Martina Patzl, Armin Saalmüller, Wilhelm Gerner
In vitro proliferation and IFN-γ production of γδ T cells. (A) Frequency of blood-derived γδ T cells from 57 six month-old pigs analysed by FCM. The box plot shows the median value of TCR-γδ+ T cells within total lymphocytes; whiskers indicate minimum and maximum values. (B–D) MACS-sorted and CellTrace™ Violet-stained blood-derived γδ T cells were stimulated for 4days with ConA, IL-2, IL-12 and IL-18 in various combinations as indicated. (B) Proliferation and IFN-γ production of γδ T cells was measured by FCM and ELISA, respectively. Numbers represent percentages of proliferating γδ T cells; black bars indicate boundary of proliferating and non-proliferating γδ T cells. Bar charts illustrate IFN-γ production in microcultures. (C) IFN-γ production of in vitro-stimulated γδ T cells from five different animals, with individuals represented by different black symbols. Black bars show the average IFN-γ production for the two combinations of stimuli. (D) Cultivated γδ T cells were stained with mAbs against CD25, CD2, SLA-DR, CD27 and CD8α. Viable cells were analysed for proliferation and the expression of cell surface markers by FCM. Grey dots within dot plots represent non-proliferating cells; green dots represent cells that have undergone one or more cycles of division. Data are representative for four individual animals analysed in four independent experiments. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.) ...In vitro proliferation and IFN-γ production of γδ T cells. (A) Frequency of blood-derived γδ T cells from 57 six month-old pigs analysed by FCM. The box plot shows the median value of TCR-γδ+ T cells within total lymphocytes; whiskers indicate minimum and maximum values. (B–D) MACS-sorted and CellTrace™ Violet-stained blood-derived γδ T cells were stimulated for 4days with ConA, IL-2, IL-12 and IL-18 in various combinations as indicated. (B) Proliferation and IFN-γ production of γδ T cells was measured by FCM and ELISA, respectively. Numbers represent percentages of proliferating γδ T cells; black bars indicate boundary of proliferating and non-proliferating γδ T cells. Bar charts illustrate IFN-γ production in microcultures. (C) IFN-γ production of in vitro-stimulated γδ T cells from five different animals, with individuals represented by different black symbols. Black bars show the average IFN-γ production for the two combinations of stimuli. (D) Cultivated γδ T cells were stained with mAbs against CD25, CD2, SLA-DR, CD27 and CD8α. Viable cells were analysed for proliferation and the expression of cell surface markers by FCM. Grey dots within dot plots represent non-proliferating cells; green dots represent cells that have undergone one or more cycles of division. Data are representative for four individual animals analysed in four independent experiments. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.) ... γδ T cells are highly abundant in the blood and spleen of pigs but little is known about their functional differentiation. In this study the potential of the type-1 polarizing cytokines IL-12 and IL-18 in combination with IL-2 and Concanavalin A (ConA) to stimulate porcine γδ T cells was investigated. Stimulation of purified γδ T cells with ConA and IL-2 induced a strong proliferation of CD2− γδ T cells, whereas additional stimulation with IL-12 and IL-18 caused a stronger proliferation of CD2+ γδ T cells. IFN-γ could only be detected in supernatants of γδ T-cell cultures supplemented with IL-12 and IL-18. Experiments with sorted CD2/SWC5-defined γδ T-cell subsets revealed that CD2+SWC5− γδ T cells are the main producers of IFN-γ following stimulation with IL-2/IL-12/IL-18. Additional stimulation with ConA led to an upregulation of CD2 within the CD2− γδ T cell subsets, indicating a previously unnoticed plasticity of CD2-defined γδ T cell subsets.