235 results for qubit oscillator frequency
Applying a field spectroscopy technique for assessing successional trends of biological soil crusts in a semi-arid environment
Contributors: E. Zaady, A. Karnieli, M. Shachak
... We studied the successional stages of biological soil crusts (BSCs) by using in-situ spectroscopic techniques during 6 years of recovery following scraping-sterilization and scraping-crumbling disturbances on north- and south-facing slopes and in plots with and without overland water runoff barriers. Two spectral indices, the Brightness Index (BI) and the Normalized Difference Vegetation Index (NDVI), were used as indicators for evaluating BSC succession, with special attention to differences between the north- and the south-facing slopes. We found that BSC succession could be expressed as linear regressions of the above-mentioned indicators during the experimental years for the different treatments. Both indicators were found to be significantly different in each of the experimental years: BI values decreased while NDVI values increased for each of the three treatments. Thus, the BI can serve as a good indicator during the early years following disturbance while the NDVI can be useful after crusts have become established. We conclude that spectral reflectance measurements of BSCs can be a useful monitoring technique for studying the regeneration of the soil surface without direct disturbance.
CRACC-targeting Fc-fusion protein induces activation of NK cells and DCs and improves T cell immune responses to antigenic targets
Contributors: Yasser A. Aldhamen, David P.W. Rastall, Weimin Chen, Sergey S. Seregin, Cristiane Pereira-Hicks, Sarah Godbehere, Norbert E. Kaminski, Andrea Amalfitano
Development of CRACC-Fc fusion protein. (A) Plasmid encoding CRACC-ECD/mIgG1-Fc was transiently transfected into 100ml suspension HEK 293-6E cell culture. CRACC-Fc protein was captured from the cell culture supernatant by HiTrapTM Mabselect 5ml column and followed by buffer exchange and SDS-PAGE and Western blot analysis. Lane M: protein marker. Lane R: reducing conditions. Lane N: non-reducing conditions. Lane P: mouse IgG1, Kappa (B and C) RAW264.7 cells (300,000cells/well) were plated in quadruplicate in 24-well plate and then stimulated for 24h with LPS (1μg/ml). After 24h, cells were harvested and incubated with escalating doses of CRACC-Fc fusion protein for 2h. After 2h, cells were then washed and stained with APC-conjugated anti-CRACC antibody and CRACC expression was evaluated by flow cytometry. (B) Representative flow cytometry figures for CRACC expression on LPS-stimulated RAW264.7 cells in the presence or absence of CRACC-Fc fusion protein. (C) Frequency of CRACC expression on LPS-stimulated RAW264.7 cells in the presence of escalating doses of CRACC-Fc fusion protein. (D) Immunoprecipitation analysis for EAT-2 expression in RAW264.7 cells following LPS stimulation in the presence or absence of CRACC-Fc (10μg/ml). (E) Quantification of EAT-2 protein levels after normalization to the 55kDa loading protein. Densitometric analysis was done using Image J software. Cells were plated in quadruplicate. The bars represent mean±SD. ...Frequency of T and B lymphocytes following rAd5-Null and CRACC-Fc co-administration. (A) Flow cytometry analysis for the percentages of CD8+ CD3+ T cells (A), CD8− CD3+ T cells (B), and CD19+ CD3− B cells (C) 12 days following rAd5-Null and either CRACC-Fc or control IgG co-administration. Statistical analysis was completed using a Student's t-test, p<0.05 was deemed as a statistically significant difference. ...CRACC-Fc fusion protein has an innate immunostimulatory activity. Male C57BL/6 mice (n=5) were i.p. injected with a mixture of rAd5-Null (2×1010vps/mouse) and 200μg of CRACC-Fc fusion protein [rAd5-Null+ CRACC-Fc) or control IgG [Ad-Null+ Control IgG]. After 12h, mice were sacrificed and splenocytes were prepared. (A) Frequency of IFNγ expressing NK cells (CD3-NK1.1+) derived from Ad-Null+ CRACC-Fc or Ad-Null+ Control IgG co-injected mice. (B and C) DCs (CD11c+, CD11b+, F4/80−) maturation was evaluated by assessing the expression levels of CD80, CD86, and I-A/I-E molecules. Expression of CD80 (B), CD86 (C), and I-A/I-E (D) on CD11c+, CD11b+, F4/80− DCs is shown. Data are presented as mean±SD. Statistical analysis was completed using a one-way analysis of variance (ANOVA) with a Student–Newman–Keuls post hoc test, p<0.05 was deemed as a statistically significant difference. *** denotes p<0.001, statistically different from naive animals. ...Cytokines secretion profiles of Gag-specific CD8+ T cells derived from rAd5-HIV/Gag and CRACC-Fc co-vaccination. Splenocytes (2×106cells/well) derived from naives (n=4) or vaccinated mice (n=7) were stimulated with the HIV-Gag immunogenic peptide, AMQMLKETI and FACS ICS analysis was performed. The Cytokines secretion profiles of HIV-Gag-specific CD8+ T cells elicited by the rAd5-HIV/Gag and CRACC-Fc co-vaccination regimen were determined by multiparameter intercellular staining assay. Gate were set based on negative control (naïve) and placed consistently across samples. The total frequency of splenic CD8+ T cells derived from naïve or vaccinated mice expressing IFNγ (A), TNFα (B), IL-2 (C), IFNγ/TNFα (D), IFNγ/IL-2 (E), or TNFα/IL-2 (F). Data are presented as mean±SD. Statistical analysis was completed using a one-way analysis of variance (ANOVA) with a Student–Newman–Keuls post hoc test, p<0.05 was deemed as a statistically significant difference. **, *** denotes p<0.01, p<0.001 statistically different from naive animals. Data are representative of three independent experiments with similar results. ...CRACC-Fc co-vaccination alters the elicited CD8+ T cells toward TEM phenotype. Phenotypic analysis of HIV-Gag-specific CD8+ T cells elicited after rAd5-HIV/Gag and CRACC-Fc co-vaccination at 42dpi. Multiparameter Gag-tetramer binding assays were performed in PBMCs (A) or splenocytes (B) of naives (n=4) or vaccinated mice (n=6) to enumerate the frequency of Gag-specific tetramer positive effector memory (CD62Llow CD127high) CD8+ T cells. Data are presented as mean±SD. Statistical analysis was completed using a one-way analysis of variance (ANOVA) with a Student–Newman–Keuls post hoc test, p<0.05 was deemed as a statistically significant difference. **, *** denotes p<0.01, p<0.001 statistically different from naive animals. Data are representative of three independent experiments with similar results. ...Increased breadth of Gag-specific CD8+ T cell responses following rAd5-HIV/Gag and CRACC-Fc co-vaccination. Splenocytes (2×106cells/well) derived from naives (n=4) or vaccinated mice (n=7) were stimulated with Gag-peptide pool# 17 (identified in Fig. 4C) and FACS ICS analysis for IFNγ-, TNFα- and IL-2-expressing CD8+ T cells was performed. The total frequency of splenic CD8+ T cells expressing IFNγ (A), TNFα (B), IL-2 (C), IFNγ/TNFα (D), IFNγ/IL-2 (E), or TNFα/IL-2 (F) are shown. Data are presented as mean±SD. Statistical analysis was completed using a one-way analysis of variance (ANOVA) with a Student–Newman–Keuls post-hoc test, p<0.05 was deemed as a statistically significant difference. **, *** denotes p<0.01, p<0.001 statistically different from naive animals. Data are representative of two independent experiments with similar results. ...CRACC-Fc fusion protein has an innate immunostimulatory activity. Male C57BL/6 mice (n=5) were i.p. injected with a mixture of rAd5-Null (2×1010vps/mouse) and 200μg of CRACC-Fc fusion protein [rAd5-Null+ CRACC-Fc) or control IgG [Ad-Null+ Control IgG]. After 12h, mice were sacrificed and splenocytes were prepared. (A) Frequency of IFNγ expressing NK cells (CD3-NK1.1+) derived from Ad-Null+ CRACC-Fc or Ad-Null+ Control IgG co-injected mice. (B and C) DCs (CD11c+, CD11b+, F4/80−) maturation was evaluated by assessing the expression levels of CD80, CD86, and I-A/I-E molecules. Expression of CD80 (B), CD86 (C), and I-A/I-E (...Increased breadth of Gag-specific CD8+ T cell responses following rAd5-HIV/Gag and CRACC-Fc co-vaccination. Splenocytes (2×106cells/well) derived from naives (n=4) or vaccinated mice (n=7) were stimulated with Gag-peptide pool# 17 (identified in Fig. 4C) and FACS ICS analysis for IFNγ-, TNFα- and IL-2-expressing CD8+ T cells was performed. The total frequency of splenic CD8+ T cells expressing IFNγ (A), TNFα (B), IL-2 (C), IFNγ/TNFα (D), IFNγ/IL-2 (E), or TNFα/IL-2 (F) are shown. Data are presented as mean±SD. Statistical analysis was completed using a one-way analysis of variance (ANOVA) with a Student–Newman–Keuls post-hoc test, psimilar results. ... The CD2-like receptor activating cytotoxic cell (CRACC) receptor is a member of the SLAM family of receptors that are found on several types of immune cells. We previously demonstrated that increasing the abundance of the adaptor protein EAT-2 during vaccination enhanced innate and adaptive immune responses to vaccine antigens. Engagement of the CRACC receptor in the presence of the EAT-2 adaptor generally results in immune cell activation, while activating CRACC signaling in cells that lack EAT-2 adaptor inhibits their effector and regulatory functions. As EAT-2 is the only SAP adaptor that interacts with the CRACC receptor, we hypothesized that technologies that specifically modulate CRACC signaling during vaccination may also improve antigen specific adaptive immune responses. To test this hypothesis, we constructed a CRACC-targeting Fc fusion protein and included it in vaccination attempts. Indeed, mice co-vaccinated with the CRACC-Fc fusion protein and an adenovirus vaccine expressing the HIV-Gag protein had improved Gag-specific T cell responses, as compared to control mice. These responses are characterized by increased numbers of Gag-specific tetramer+ CD8+ T cells and increases in production of IFNγ, TNFα, and IL2, by Gag-specific CD8+ T cells. Moreover, our results revealed that use of the CRACC-Fc fusion protein enhances vaccine-elicited innate immune responses, as characterized by increased dendritic cells (DCs) maturation and IFNγ production from NK cells. This study highlights the importance of CRACC signaling during the induction of an immune response generally, and during vaccinations specifically, and also lends insight into the mechanisms underlying our prior results noting EAT-2-dependent improvements in vaccine efficacy.
Contributors: Morteza Fattahi, Richard T. Walker
... Iran is one of the world's most tectonically active regions, yet dating past earthquakes for neotectonic studies has been limited. One of the main reasons for this is that organic material suitable for radiocarbon dating of deformed sediments is rare. We investigate the use of infrared stimulated luminescence (IRSL) from coarse-grained feldspars to date colluvial deposits associated with the Sabzevar thrust fault in northeastern Iran. The single-aliquot regenerative (SAR) dose measurement procedure was used for this study. The current study investigates monitoring and correcting for sensitivity changes, recovering a known laboratory dose and equivalent dose estimation using three SAR IRSL methods. It is shown that SAR has recovered a given laboratory dose using a range of preheat temperatures but De determination of natural samples requires its own preheat plateaus for two of these SAR methods. The SAR IRSL method provided an age of 1.7±0.3ka for colluvium, predating the last earthquake event on the Sabzevar fault. This result suggests that this fault is likely to be responsible for an earthquake that destroyed Sabzevar city in AD 1052.
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.
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.