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Sample records for advisories targeting sensitive

  1. Cost Sensitive Moving Target Consensus

    SciTech Connect

    Duan, Sisi; Li, Yun; Levitt, Karl N.

    2016-01-01

    Consensus is a fundamental approach to implementing fault-tolerant services through replication where there exists a tradeoff between the cost and the resilience. For instance, Crash Fault Tolerant (CFT) protocols have a low cost but can only handle crash failures while Byzantine Fault Tolerant (BFT) protocols handle arbitrary failures but have a higher cost. Hybrid protocols enjoy the benefits of both high performance without failures and high resiliency under failures by switching among different subprotocols. However, it is challenging to determine which subprotocols should be used. We propose a moving target approach to switch among protocols according to the existing system and network vulnerability. At the core of our approach is a formalized cost model that evaluates the vulnerability and performance of consensus protocols based on real-time Intrusion Detection System (IDS) signals. Based on the evaluation results, we demonstrate that a safe, cheap, and unpredictable protocol is always used and a high IDS error rate can be tolerated.

  2. Temperature targets revisited under climate sensitivity uncertainty

    NASA Astrophysics Data System (ADS)

    Neubersch, Delf; Roth, Robert; Held, Hermann

    2015-04-01

    While the 2° target has become an official goal of the COP (Conference of the Parties) process recent work has shown that it requires re-interpretation if climate sensitivity uncertainty in combination with anticipated future learning is considered (Schmidt et al., 2011). A strict probabilistic limit as suggested by the Copenhagen diagnosis may lead to conceptual flaws in view of future learning such a negative expected value of information or even ill-posed policy recommendations. Instead Schmidt et al. suggest trading off the probabilistic transgression of a temperature target against mitigation-induced welfare losses and call this procedure cost risk analysis (CRA). Here we spell out CRA for the integrated assessment model MIND and derive necessary conditions for the exact nature of that trade-off. With CRA at hand it is for the first time that the expected value of climate information, for a given temperature target, can meaningfully be assessed. When focusing on a linear risk function as the most conservative of all possible risk functions, we find that 2° target-induced mitigation costs could be reduced by up to 1/3 if the climate response to carbon dioxide emissions were known with certainty, amounting to hundreds of billions of Euros per year (Neubersch et al., 2014). Further benefits of CRA over strictly formulated temperature targets are discussed. References: D. Neubersch, H. Held, A. Otto, Operationalizing climate targets under learning: An application of cost-risk analysis, Climatic Change, 126 (3), 305-318, DOI 10.1007/s10584-014-1223-z (2014). M. G. W. Schmidt, A. Lorenz, H. Held, E. Kriegler, Climate Targets under Uncertainty: Challenges and Remedies, Climatic Change Letters, 104 (3-4), 783-791, DOI 10.1007/s10584-010-9985-4 (2011).

  3. Continuous psychophysics: Target-tracking to measure visual sensitivity

    PubMed Central

    Bonnen, Kathryn; Burge, Johannes; Yates, Jacob; Pillow, Jonathan; Cormack, Lawrence K.

    2015-01-01

    We introduce a novel framework for estimating visual sensitivity using a continuous target-tracking task in concert with a dynamic internal model of human visual performance. Observers used a mouse cursor to track the center of a two-dimensional Gaussian luminance blob as it moved in a random walk in a field of dynamic additive Gaussian luminance noise. To estimate visual sensitivity, we fit a Kalman filter model to the human tracking data under the assumption that humans behave as Bayesian ideal observers. Such observers optimally combine prior information with noisy observations to produce an estimate of target position at each time step. We found that estimates of human sensory noise obtained from the Kalman filter fit were highly correlated with traditional psychophysical measures of human sensitivity (R2 > 97%). Because each frame of the tracking task is effectively a “minitrial,” this technique reduces the amount of time required to assess sensitivity compared with traditional psychophysics. Furthermore, because the task is fast, easy, and fun, it could be used to assess children, certain clinical patients, and other populations that may get impatient with traditional psychophysics. Importantly, the modeling framework provides estimates of decision variable variance that are directly comparable with those obtained from traditional psychophysics. Further, we show that easily computed summary statistics of the tracking data can also accurately predict relative sensitivity (i.e., traditional sensitivity to within a scale factor). PMID:25795437

  4. Architecture Considerations of Time Sensitive Targeting and NATO TST Tool

    DTIC Science & Technology

    2013-06-01

    Architecture Considerations of Time Sensitive Targeting and NATO TST Tool Orhan Cetinkaya, PhD Yakup Yildirim, PhD NCI Agency, 19-21 June 2013...SAR) 18. NUMBER OF PAGES 18 19a. NAME OF RESPONSIBLE PERSON a. REPORT unclassified b . ABSTRACT unclassified c. THIS PAGE unclassified...8217’_’ _’ •_· __ ~==============================================~--0-~ __ is-ta_r_~ ~ NATO UNCLASSIFIED 18 Questions? Orhan Cetinkaya, PhD Principal Scientist

  5. Adaptive target detection with a polarization-sensitive optical system.

    PubMed

    Meng, Lingfei; Kerekes, John P

    2011-05-01

    We developed an adaptive polarimetric target detector (APTD) to determine the optimum combination strategy for a multichannel polarization-sensitive optical system. The proposed algorithm is based on scene-derived polarization properties of the target and background, and it seeks to find an optimum multichannel combination of linear polarizing filters that maximizes the signal-to-clutter ratio (SCR) in intensity and Stokes parameter images. The algorithm is validated by performing RX anomaly detection and a generalized likelihood ratio test on both synthetic and real imagery. The experimental results are analyzed through calculated SCR and receiver operating characteristic curves. Compared with several conventional operation methods, we find that better target detection performance is achieved with the APTD algorithm.

  6. Contact dermatitis: in pursuit of sensitizer's molecular targets through proteomics.

    PubMed

    Guedes, Sofia; Neves, Bruno; Vitorino, Rui; Domingues, Rosário; Cruz, Maria Teresa; Domingues, Pedro

    2017-02-01

    Protein haptenation, i.e., the modification of proteins by small reactive chemicals, is the key step in the sensitization phase of allergic contact dermatitis (ACD). Despite the research effort in past decades, the identification of immunogenic hapten-protein complexes that trigger a relevant pathogenic immune response in ACD, as well as the haptenation reaction molecular site, and the elements of a potentially conditioning environment during each of these stages, remain poorly understood. These questions led us to employ a proteomics-based approach to identify modified proteins in the dendritic-like cell line THP-1 sensitized with fluorescein isothiocyanate (FITC), through a combination of 2D-gel electrophoresis, nano-LC and mass spectrometry. A specific set of 39 targeted proteins was identified and comprised proteins from various cellular locations and biological functions. One of FITC targets was identified as MLK, a member of the mixed-lineage kinase family known to act as a mitogen-activated protein kinase kinase kinase and to control the activity of specific mitogen-activated protein kinase pathways, namely p38 and JNK pathways. Haptenated in the vicinity of its active site, our results point to MLK being a relevant target due to a consistent non-activation at early time points of these pathways upon FITC sensitization in THP-1 cells. Moreover, FITC pre-treatment significantly decrease phospho-p38 and phospho-JNK levels induced upon exposure to a classical activator such as lipopolysaccharide or to the sensitizer 2,4-dinitrofluorobenzene. Overall, our data point to specific amino acid residues haptenation within critical proteins as the key step in the subsequent signaling pathways modulation responsible for DC activation and maturation events.

  7. Targeting Smoothened Sensitizes Gastric Cancer to Chemotherapy in Experimental Models

    PubMed Central

    Ma, Huifa; Tian, Yongsheng; Yu, Xiangyang

    2017-01-01

    Background The Hedgehog pathway receptor smoothened (SMO) has critical roles in tumor progression. However, whether SMO is a key factor regulating gastric cancer chemotherapy resistance is unknown. Material/Methods We investigated the potential functions of SMO in inducing gastric cancer paclitaxel resistance in clinical samples, gastric cancer cell lines (424GC and AGS), and subcutaneous syngeneic mouse models. Results We found high SMO expression in paclitaxel-resistant gastric cancer clinical samples. Paclitaxel gastric cancer cells had higher SMO expression than in drug-sensitive cells. Upregulating SMO expression induced paclitaxel resistance in gastric cells lines via enhancing cell proliferation and inhibiting apoptosis. The combination of IPI-926, an inhibitor of SMO, with paclitaxel decreased cell viability of paclitaxel-resistant gastric cancer cells in vitro and controlled tumor growth in animal models. Conclusions The Hedgehog pathway receptor SMO is an important regulator of gastric cancer paclitaxel resistance and could be a target for sensitizing paclitaxel-resistant tumors. PMID:28350784

  8. Silent Warning: Understanding the National Terrorism Advisory System

    DTIC Science & Technology

    2014-12-01

    methodology for the issuance and revocation of advisories; develop the content of these advisories; develop targeted advisories for specific regions...of HSPD-3, in that DHS was directed to establish criteria and methodology for the issuance and revocation of advisories; develop the content of...the issuance and revocation of advisories; develop the content of these advisories; develop targeted advisories for specific regions, localities

  9. Targeting Nucleophosmin 1 Represents a Rational Strategy for Radiation Sensitization

    SciTech Connect

    Sekhar, Konjeti R.; Benamar, Mouadh; Venkateswaran, Amudhan; Sasi, Soumya; Penthala, Narsimha R.; Crooks, Peter A.; Hann, Stephen R.; Geng, Ling; Balusu, Ramesh; Abbas, Tarek; Freeman, Michael L.

    2014-08-01

    Purpose: To test the hypothesis that small molecule targeting of nucleophosmin 1 (NPM1) represents a rational approach for radiosensitization. Methods and Materials: Wilde-type and NPM1-deficient mouse embryo fibroblasts (MEFs) were used to determine whether radiosensitization produced by the small molecule YTR107 was NPM1 dependent. The stress response to ionizing radiation was assessed by quantifying pNPM1, γH2AX, and Rad51 foci, neutral comet tail moment, and colony formation. NPM1 levels in a human-derived non-small-cell lung cancer (NSCLC) tissue microarray (TMA) were determined by immunohistochemistry. YTR107-mediated radiosensitization was assessed in NSCLC cell lines and xenografts. Results: Use of NPM1-null MEFs demonstrated that NPM1 is critical for DNA double- strand break (DSB) repair, that loss of NPM1 increases radiation sensitivity, and that YTR107-mediated radiosensitization is NPM1 dependent. YTR107 was shown to inhibit NPM1 oligomerization and impair formation of pNPM1 irradiation-induced foci that colocalized with γH2AX foci. Analysis of the TMA demonstrated that NPM1 is overexpressed in subsets of NSCLC. YTR107 inhibited DNA DSB repair and radiosensitized NSCLC lines and xenografts. Conclusions: These data demonstrate that YTR107-mediated targeting of NPM1 impairs DNA DSB repair, an event that increases radiation sensitivity.

  10. Particulate matter inflammation and receptor sensitivity are target cell specific.

    PubMed

    Veronesi, Bellina; de Haar, Colin; Roy, Josee; Oortgiesen, Marga

    2002-02-01

    The complexity of primary source particulate matter (PM) and the various cell types encountered by its inhalation raise the possibility that target cells are differentially activated. Since epithelial cells, which line the nasal-tracheal-bronchial airways, and sensory C fibers, which terminate throughout this epithelial layer, are initially targeted by inhaled PM, we compared their relative biological response in vitro to PM originating from volcanic (MSH), anthropogenic (diesel), residential (woodstove), urban ambient (St. Louis, Ottawa), and industrial emission (coal fly ash, CFA; residual oil fly ash, ROFA; oil fly ash, OFA) sources. Increases in intracellular calcium (i.e., [Ca(2+)](i)) are a second-messenger event that indicates cellular activation and signal transduction, in both nerve and epithelial cells. Single-cell calcium imaging recordings were taken of human bronchial epithelial cells (BEAS-2B) exposed to selected PM (50 microg/ml or 30 microg/cm(2)). These cells responded with variable increases in [Ca(2+)](i) ranging from abrupt increases, which returned to baseline upon washing of the cells, to oscillations of the [Ca(2+)](i) that did not wash out. Increases in [Ca(2+)](i) and inflammatory cytokine (i.e., interleukin 6, IL-6) release were measured in populations of BEAS-2B cells exposed to PM (50 microg/ml) and were shown to significantly correlate (r(2) =.80). BEAS-2B cells, stained histochemically with cobalt, displayed a concentration-dependent precipitation in response to acid pH and capsaicin, indicating the presence of acid-sensitive pathways (e.g., VR1 and acid-sensitive receptors). To demonstrate the relevance of these pathways to inflammatory cytokine (i.e., IL-6) release, BEAS-2B cells were pretreated (15 min) with antagonists to the vanilloid (VR1) receptor (i.e., capsazepine, CPZ) or acid-sensitive pathways (i.e., amiloride) before their exposure to the selected PM. A significant reduction of IL-6 release occurred in response to all PM

  11. Evaluating risk communication: examining target audience perceptions about four presentation formats for fish consumption health advisory information.

    PubMed

    Connelly, N A; Knuth, B A

    1998-10-01

    Information format can influence the extent to which target audiences understand and respond to risk-related information. This study examined four elements of risk information presentation format. Using printed materials, we examined target audience perceptions about: (a) reading level; (b) use of diagrams vs. text; (c) commanding versus cajoling tone; and (d) use of qualitative vs. quantitative information presented in a risk ladder. We used the risk communication topic of human health concerns related to eating noncommercial Great Lakes fish affected by chemical contaminants. Results from the comparisons of specific communication formats indicated that multiple formats are required to meet the needs of a significant percent of anglers for three of the four format types examined. Advisory text should be reviewed to ensure the reading level is geared to abilities of the target audience. For many audiences, a combination of qualitative and quantitative information, and a combination of diagrams and text may be most effective. For most audiences, a cajoling rather than commanding tone better provides them with the information they need to make a decision about fish consumption. Segmenting audiences regarding information needs and communication formats may help clarify which approaches to take with each audience.

  12. An attempt to target anxiety sensitivity via cognitive bias modification.

    PubMed

    Clerkin, Elise M; Beard, Courtney; Fisher, Christopher R; Schofield, Casey A

    2015-01-01

    Our goals in the present study were to test an adaptation of a Cognitive Bias Modification program to reduce anxiety sensitivity, and to evaluate the causal relationships between interpretation bias of physiological cues, anxiety sensitivity, and anxiety and avoidance associated with interoceptive exposures. Participants with elevated anxiety sensitivity who endorsed having a panic attack or limited symptom attack were randomly assigned to either an Interpretation Modification Program (IMP; n = 33) or a Control (n = 32) condition. During interpretation modification training (via the Word Sentence Association Paradigm), participants read short sentences describing ambiguous panic-relevant physiological and cognitive symptoms and were trained to endorse benign interpretations and reject threatening interpretations associated with these cues. Compared to the Control condition, IMP training successfully increased endorsements of benign interpretations and decreased endorsements of threatening interpretations at visit 2. Although self-reported anxiety sensitivity decreased from pre-selection to visit 1 and from visit 1 to visit 2, the reduction was not larger for the experimental versus control condition. Further, participants in IMP (vs. Control) training did not experience less anxiety and avoidance associated with interoceptive exposures. In fact, there was some evidence that those in the Control condition experienced less avoidance following training. Potential explanations for the null findings, including problems with the benign panic-relevant stimuli and limitations with the control condition, are discussed.

  13. Targeting drug sensitivity predictors: New potential strategies to improve pharmacotherapy of human brain disorders.

    PubMed

    Kalueff, Allan V; Stewart, Adam Michael; Nguyen, Michael; Song, Cai; Gottesman, Irving I

    2015-12-03

    One of the main challenges in medicine is the lack of efficient drug therapies for common human disorders. For example, although depressed patients receive powerful antidepressants, many often remain resistant to psychopharmacotherapy. The growing recognition of complex interplay between the drug targets and the predictors of drug sensitivity requires an improved understanding of these two key aspects of drug action and their potentially shared molecular networks. Here, we apply the concept of endophenotypes and their interplay to drug action and sensitivity. Based on these analyses, we postulate that novel drugs may be developed by targeting specific molecular pathways that integrate drug targets with drug sensitivity predictors.

  14. Targeting Breast Cancer Recurrence via Hedgehog-mediated Sensitization of Breast Cancer Stem Cells

    DTIC Science & Technology

    2011-07-01

    established a treatment model in which the Smoothened agonist was sufficient to partially rescue the Sonic Hedgehog knockout suggesting that similar... Hedgehog -mediated Sensitization of Breast Cancer Stem Cells PRINCIPAL INVESTIGATOR: David J. Robbins, Ph.D...June 2010 – 14 June 2011 4. TITLE AND SUBTITLE 5a. CONTRACT NUMBER Targeting Breast Cancer Recurrence via Hedgehog -mediated Sensitization of

  15. Hearing sensitivity during target presence and absence while a whale echolocates.

    PubMed

    Supin, Alexander Ya; Nachtigall, Paul E; Breese, Marlee

    2008-01-01

    Hearing sensitivity was measured in a false killer whale during echolocation. Sensitivity was measured using probe stimuli as sinusoidally amplitude modulated signals with a 22.5-kHz carrier frequency and recording auditory evoked potentials as envelope-following responses. The probes were presented and responses were recorded during short 2-s periods when the animal echolocated to detect the presence or absence of a target in a go/no-go paradigm. In the target-absent trials, a hearing threshold of 90.4 dB re 1 muPa was found; in the target-present trials, the threshold was 109.8 dB. Thus, a 19.4-dB difference was found between thresholds in the target-present and target-absent trials. To check the possibility that this difference was the result of different masking degree of the probe by the emitted sonar clicks, click statistics were investigated in similar trials. No indication was found that the energy of the emitted clicks was higher in the target-present than in target-absent trials; on the contrary, mean click level, mean number of clicks per train, and overall train energy was slightly higher in the target-absent trials. Thus the data indicate that the hearing sensitivity of the whale varied depending on target presence or absence.

  16. Identification of cellular targets of a series of boron heterocycles using TIPA II-A sensitive target identification platform.

    PubMed

    Ward, Matthew S; Silva, Isba; Martinez, Walfre; Jefferson, Jameka; Rahman, Shakila; Garcia, Jeanie M; Kanichar, Divya; Roppiyakuda, Lance; Kosmowska, Ewa; Faust, Michelle A; Tran, Kim P; Chow, Felicia; Buglo, Elena; Zhou, Feimeng; Groziak, Michael P; Xu, H Howard

    2016-08-01

    One of the hurdles in the discovery of antibiotics is the difficulty of linking antibacterial compounds to their cellular targets. Our laboratory has employed a genome-wide approach of over-expressing essential genes in order to identify cellular targets of antibacterial inhibitors. Our objective in this project was to develop and validate a more sensitive disk diffusion based platform of target identification (Target Identification Platform for Antibacterials version 2; TIPA II) using a collection of cell clones in an Escherichia coli mutant (AS19) host with increased outer membrane permeability. Five known antibiotics/inhibitors and 28 boron heterocycles were tested by TIPA II assay, in conjunction with the original assay TIPA. The TIPA II was more sensitive than TIPA because eight boron heterocycles previously found to be inactive to AG1 cells in TIPA assays exhibited activity to AS19 cells. For 15 boron heterocycles, resistant colonies were observed within the zones of inhibition only on the inducing plates in TIPA II assays. DNA sequencing confirmed that resistant clones harbor plasmids with fabI gene as insert, indicating that these boron heterocycles all target enoyl ACP reductase. Additionally, cell-based assays and dose response curved obtained indicated that for two boron heterocycle inhibitors, the fabI cell clone in AG1 (wild-type) host cells exhibited at least 11 fold more resistance under induced conditions than under non-induced conditions. Moreover, TIPA II also identified cellular targets of known antibacterial inhibitors triclosan, phosphomycin, trimethoprim, diazaborine and thiolactomycin, further validating the utility of the new system.

  17. Targeting Breast Cancer Recurrence via Hedgehog-Mediated Sensitization of Breast Cancer Stem Cells

    DTIC Science & Technology

    2011-07-01

    force these cells back into the cell cycle, possibly sensitizing them to adjuvant cancer therapeutics. To test this hypothesis we developed three...force these cells back into the cell cycle, possibly sensitizing them to adjuvant cancer therapeutics. To test this hypothesis we developed three...canonical hedgehog target gene, Patched1, was not altered in response to oral-gavage of Smoothened agonists. In vitro testing of the compounds

  18. Differential Sensitivity of Target Genes to Translational Repression by miR-17~92

    PubMed Central

    Jin, Hyun Yong; Oda, Hiroyo; Chen, Pengda; Kang, Seung Goo; Valentine, Elizabeth; Liao, Lujian; Zhang, Yaoyang; Gonzalez-Martin, Alicia; Shepherd, Jovan; Head, Steven R.; Kim, Pyeung-Hyeun; Fu, Guo; Liu, Wen-Hsien; Han, Jiahuai

    2017-01-01

    MicroRNAs (miRNAs) are thought to exert their functions by modulating the expression of hundreds of target genes and each to a small degree, but it remains unclear how small changes in hundreds of target genes are translated into the specific function of a miRNA. Here, we conducted an integrated analysis of transcriptome and translatome of primary B cells from mutant mice expressing miR-17~92 at three different levels to address this issue. We found that target genes exhibit differential sensitivity to miRNA suppression and that only a small fraction of target genes are actually suppressed by a given concentration of miRNA under physiological conditions. Transgenic expression and deletion of the same miRNA gene regulate largely distinct sets of target genes. miR-17~92 controls target gene expression mainly through translational repression and 5’UTR plays an important role in regulating target gene sensitivity to miRNA suppression. These findings provide molecular insights into a model in which miRNAs exert their specific functions through a small number of key target genes. PMID:28241004

  19. Long-Term Memories Bias Sensitivity and Target Selection in Complex Scenes

    PubMed Central

    Patai, Eva Zita; Doallo, Sonia; Nobre, Anna Christina

    2014-01-01

    In everyday situations we often rely on our memories to find what we are looking for in our cluttered environment. Recently, we developed a new experimental paradigm to investigate how long-term memory (LTM) can guide attention, and showed how the pre-exposure to a complex scene in which a target location had been learned facilitated the detection of the transient appearance of the target at the remembered location (Summerfield, Lepsien, Gitelman, Mesulam, & Nobre, 2006; Summerfield, Rao, Garside, & Nobre, 2011). The present study extends these findings by investigating whether and how LTM can enhance perceptual sensitivity to identify targets occurring within their complex scene context. Behavioral measures showed superior perceptual sensitivity (d′) for targets located in remembered spatial contexts. We used the N2pc event-related potential to test whether LTM modulated the process of selecting the target from its scene context. Surprisingly, in contrast to effects of visual spatial cues or implicit contextual cueing, LTM for target locations significantly attenuated the N2pc potential. We propose that the mechanism by which these explicitly available LTMs facilitate perceptual identification of targets may differ from mechanisms triggered by other types of top-down sources of information. PMID:23016670

  20. Secure FAST: Security Enhancement in the NATO Time Sensitive Targeting Tool

    DTIC Science & Technology

    2010-11-01

    RTO-MP-IST-091 P9 - 1 Secure FAST: Security Enhancement in the NATO Time Sensitive Targeting Tool Dr Orhan Cetinkaya, Dr Yakup Yildirim...OF: 17. LIMITATION OF ABSTRACT SAR 18. NUMBER OF PAGES 6 19a. NAME OF RESPONSIBLE PERSON a. REPORT unclassified b . ABSTRACT unclassified c

  1. Target-protecting dumbbell molecular probe against exonucleases digestion for sensitive detection of ATP and streptavidin.

    PubMed

    Chen, Jinyang; Liu, Yucheng; Ji, Xinghu; He, Zhike

    2016-09-15

    In this work, a versatile dumbbell molecular (DM) probe was designed and employed in the sensitively homogeneous bioassay. In the presence of target molecule, the DM probe was protected from the digestion of exonucleases. Subsequently, the protected DM probe specifically bound to the intercalation dye and resulted in obvious fluorescence signal which was used to determine the target molecule in return. This design allows specific and versatile detection of diverse targets with easy operation and no sophisticated fluorescence labeling. Integrating the idea of target-protecting DM probe with adenosine triphosphate (ATP) involved ligation reaction, the DM probe with 5'-end phosphorylation was successfully constructed for ATP detection, and the limitation of detection was found to be 4.8 pM. Thanks to its excellent selectivity and sensitivity, this sensing strategy was used to detect ATP spiked in human serum as well as cellular ATP. Moreover, the proposed strategy was also applied in the visual detection of ATP in droplet-based microfluidic platform with satisfactory results. Similarly, combining the principle of target-protecting DM probe with streptavidin (SA)-biotin interaction, the DM probe with 3'-end biotinylation was developed for selective and sensitive SA determination, which demonstrated the robustness and versatility of this design.

  2. Highly sensitive and specific colorimetric detection of cancer cells via dual-aptamer target binding strategy.

    PubMed

    Wang, Kun; Fan, Daoqing; Liu, Yaqing; Wang, Erkang

    2015-11-15

    Simple, rapid, sensitive and specific detection of cancer cells is of great importance for early and accurate cancer diagnostics and therapy. By coupling nanotechnology and dual-aptamer target binding strategies, we developed a colorimetric assay for visually detecting cancer cells with high sensitivity and specificity. The nanotechnology including high catalytic activity of PtAuNP and magnetic separation & concentration plays a vital role on the signal amplification and improvement of detection sensitivity. The color change caused by small amount of target cancer cells (10 cells/mL) can be clearly distinguished by naked eyes. The dual-aptamer target binding strategy guarantees the detection specificity that large amount of non-cancer cells and different cancer cells (10(4) cells/mL) cannot cause obvious color change. A detection limit as low as 10 cells/mL with detection linear range from 10 to 10(5) cells/mL was reached according to the experimental detections in phosphate buffer solution as well as serum sample. The developed enzyme-free and cost effective colorimetric assay is simple and no need of instrument while still provides excellent sensitivity, specificity and repeatability, having potential application on point-of-care cancer diagnosis.

  3. Dependence On Glycolysis Sensitizes BRAF-mutated Melanomas For Increased Response To Targeted BRAF Inhibition

    PubMed Central

    Hardeman, Keisha N.; Peng, Chengwei; Paudel, Bishal B.; Meyer, Christian T.; Luong, Thong; Tyson, Darren R.; Young, Jamey D.; Quaranta, Vito; Fessel, Joshua P.

    2017-01-01

    Dysregulated metabolism can broadly affect therapy resistance by influencing compensatory signaling and expanding proliferation. Given many BRAF-mutated melanoma patients experience disease progression with targeted BRAF inhibitors, we hypothesized therapeutic response is related to tumor metabolic phenotype, and that altering tumor metabolism could change therapeutic outcome. We demonstrated the proliferative kinetics of BRAF-mutated melanoma cells treated with the BRAF inhibitor PLX4720 fall along a spectrum of sensitivity, providing a model system to study the interplay of metabolism and drug sensitivity. We discovered an inverse relationship between glucose availability and sensitivity to BRAF inhibition through characterization of metabolic phenotypes using nearly a dozen metabolic parameters in Principle Component Analysis. Subsequently, we generated rho0 variants that lacked functional mitochondrial respiration and increased glycolytic metabolism. The rho0 cell lines exhibited increased sensitivity to PLX4720 compared to the respiration-competent parental lines. Finally, we utilized the FDA-approved antiretroviral drug zalcitabine to suppress mitochondrial respiration and to force glycolysis in our cell line panel, resulting in increased PLX4720 sensitivity via shifts in EC50 and Hill slope metrics. Our data suggest that forcing tumor glycolysis in melanoma using zalcitabine or other similar approaches may be an adjunct to increase the efficacy of targeted BRAF therapy. PMID:28205616

  4. Target distance-dependent variation of hearing sensitivity during echolocation in a false killer whale.

    PubMed

    Supin, Alexander Ya; Nachtigall, Paul E; Breese, Marlee

    2010-06-01

    Evidence of varying hearing sensitivity according to the target distance was obtained in a false killer whale Pseudorca crassidens during echolocation. Auditory evoked potentials (AEPs) triggered by echolocation clicks were recorded. The target distance varied from 1 to 6 m. The records contained AEPs to the self-heard emitted click and AEPs to the echoes. Mean level of echolocation clicks depended on distance (the longer the distance, the higher the click level), however, the effect of click level on AEP amplitude was eliminated by extracting AEPs to clicks of certain particular levels. The amplitude of the echo-provoked AEP was almost independent of distance, however, the amplitude of the AEP to the emitted click, did depend on distance within a range from 1 to 4 m: the longer the distance, the higher the amplitude. The latter result is interpreted as confirmational evidence that the animal is capable of varying hearing sensitivity according to target distance. The variation of hearing sensitivity may help to compensate for the echo attenuation with distance; as a secondary effect, this variation manifested itself in a variation of the amplitude of the AEP to emitted clicks.

  5. Development and characterization of site specific target sensitive liposomes for the delivery of thrombolytic agents.

    PubMed

    Vaidya, Bhuvaneshwar; Nayak, Manasa K; Dash, Debabrata; Agrawal, G P; Vyas, Suresh P

    2011-01-17

    In recent times, search for potent and highly selective thrombolytic agents with minimal side effects has become a major area of research. The aim of the present study was to develop and characterize target sensitive (TS) liposomes encapsulating streptokinase, a thrombolytic agent. The developed TS liposomes were composed of dioleylphophatidyl ethanolamine (DOPE) and dipalmityl-c(RGDfK) (10:1mol/mol). Dipalmityl-c(RGDfK) was synthesized using typical carbodiimide chemistry using palmitic acid and c(RGDfK), while lysine was used as a spacer. Liposomes were of 100-120nm size. In vitro drug release study showed that nearly 40% drug of the entrapped drug was released in 12h in the PBS (pH 7.4), however on incubation with activated platelet about 90% of drug was released within 45min. The results suggested target sensitivity of the liposomes. Further, targeting potential was confirmed using fluorescent microscopy and flow cytometry. Clot lysis study revealed that TS liposomes could not only reduce the clot lysis time but also increase the extent of clot lysis as compared to non-liposomal streptokinase solution. In conclusion, the present liposomal formulation will target the thrombolytic agent to the activated platelets in the thrombus and hence will improve the therapeutic efficacy of the drug.

  6. A Genetic Screen Reveals Novel Targets to Render Pseudomonas aeruginosa Sensitive to Lysozyme and Cell Wall-Targeting Antibiotics

    PubMed Central

    Lee, Kang-Mu; Lee, Keehoon; Go, Junhyeok; Park, In Ho; Shin, Jeon-Soo; Choi, Jae Young; Kim, Hyun Jik; Yoon, Sang Sun

    2017-01-01

    Pseudomonas aeruginosa is capable of establishing airway infections. Human airway mucus contains a large amount of lysozyme, which hydrolyzes bacterial cell walls. P. aeruginosa, however, is known to be resistant to lysozyme. Here, we performed a genetic screen using a mutant library of PAO1, a prototype P. aeruginosa strain, and identified two mutants (ΔbamB and ΔfabY) that exhibited decrease in survival after lysozyme treatment. The bamB and fabY genes encode an outer membrane assembly protein and a fatty acid synthesis enzyme, respectively. These two mutants displayed retarded growth in the airway mucus secretion (AMS). In addition, these mutants exhibited reduced virulence and compromised survival fitness in two different in vivo infection models. The mutants also showed susceptibility to several antibiotics. Especially, ΔbamB mutant was very sensitive to vancomycin, ampicillin, and ceftazidime that target cell wall synthesis. The ΔfabY displayed compromised membrane integrity. In conclusion, this study uncovered a common aspect of two different P. aeruginosa mutants with pleiotropic phenotypes, and suggests that BamB and FabY could be novel potential drug targets for the treatment of P. aeruginosa infection. PMID:28299285

  7. A Genetic Screen Reveals Novel Targets to Render Pseudomonas aeruginosa Sensitive to Lysozyme and Cell Wall-Targeting Antibiotics.

    PubMed

    Lee, Kang-Mu; Lee, Keehoon; Go, Junhyeok; Park, In Ho; Shin, Jeon-Soo; Choi, Jae Young; Kim, Hyun Jik; Yoon, Sang Sun

    2017-01-01

    Pseudomonas aeruginosa is capable of establishing airway infections. Human airway mucus contains a large amount of lysozyme, which hydrolyzes bacterial cell walls. P. aeruginosa, however, is known to be resistant to lysozyme. Here, we performed a genetic screen using a mutant library of PAO1, a prototype P. aeruginosa strain, and identified two mutants (ΔbamB and ΔfabY) that exhibited decrease in survival after lysozyme treatment. The bamB and fabY genes encode an outer membrane assembly protein and a fatty acid synthesis enzyme, respectively. These two mutants displayed retarded growth in the airway mucus secretion (AMS). In addition, these mutants exhibited reduced virulence and compromised survival fitness in two different in vivo infection models. The mutants also showed susceptibility to several antibiotics. Especially, ΔbamB mutant was very sensitive to vancomycin, ampicillin, and ceftazidime that target cell wall synthesis. The ΔfabY displayed compromised membrane integrity. In conclusion, this study uncovered a common aspect of two different P. aeruginosa mutants with pleiotropic phenotypes, and suggests that BamB and FabY could be novel potential drug targets for the treatment of P. aeruginosa infection.

  8. Anxiety Sensitivity Amelioration Training (ASAT): a longitudinal primary prevention program targeting cognitive vulnerability.

    PubMed

    Schmidt, Norman B; Eggleston, A Meade; Woolaway-Bickel, Kelly; Fitzpatrick, Kathleen Kara; Vasey, Michael W; Richey, J Anthony

    2007-01-01

    Fear of arousal symptoms, often referred to as anxiety sensitivity (AS) appears to be associated with risk for anxiety pathology and other Axis I conditions. Findings from a longitudinal prevention program targeting AS are reported. Participants (n=404) scoring high on the Anxiety Sensitivity Index (ASI) were randomly assigned to receive a brief intervention designed to reduce AS (Anxiety Sensitivity Amelioration Training (ASAT)) or a control condition. Participants were followed for up to 24 months. Findings indicate that ASAT produced greater reductions in ASI levels compared with the control condition. Moreover, reductions were specific to anxiety sensitivity relative to related cognitive risk factors for anxiety. ASAT also produced decreased subjective fear responding to a 20% CO(2) challenge delivered postintervention. Data from the follow-up period show a lower incidence of Axis I diagnoses in the treated condition though the overall group difference was not statistically different at all follow-up intervals. Overall, findings are promising for the preventative efficacy of a brief, computer-based intervention designed to decrease anxiety sensitivity.

  9. Highly Sensitive Detection of Target Biomolecules on Cell Surface Using Gold Nanoparticle Conjugated with Aptamer Probe

    NASA Astrophysics Data System (ADS)

    Kim, Hyonchol; Terazono, Hideyuki; Hayashi, Masahito; Takei, Hiroyuki; Yasuda, Kenji

    2012-06-01

    A method of gold nanoparticle (Au NP) labeling with backscattered electron (BE) imaging of field emission scanning electron microscopy (FE-SEM) was applied for specific detection of target biomolecules on a cell surface. A single-stranded DNA aptamer, which specifically binds to the target molecule on a human acute lymphoblastic leukemia cell, was conjugated with a 20 nm Au NP and used as a probe to label its target molecule on the cell. The Au NP probe was incubated with the cell, and the interaction was confirmed using BE imaging of FE-SEM through direct counting of the number of Au NPs attached on the target cell surface. Specific Au NP-aptamer probes were observed on a single cell surface and their spatial distributions including submicron-order localizations were also clearly visualized, whereas the nonspecific aptamer probes were not observed on it. The aptamer probe can be potentially dislodged from the cell surface with treatment of nucleases, indicating that Au NP-conjugated aptamer probes can be used as sensitive and reversible probes to label target biomolecules on cells.

  10. Advancing the sensitivity of selected reaction monitoring-based targeted quantitative proteomics

    SciTech Connect

    Shi, Tujin; Su, Dian; Liu, Tao; Tang, Keqi; Camp, David G.; Qian, Weijun; Smith, Richard D.

    2012-04-01

    Selected reaction monitoring (SRM)—also known as multiple reaction monitoring (MRM)—has emerged as a promising high-throughput targeted protein quantification technology for candidate biomarker verification and systems biology applications. A major bottleneck for current SRM technology, however, is insufficient sensitivity for e.g., detecting low-abundance biomarkers likely present at the pg/mL to low ng/mL range in human blood plasma or serum, or extremely low-abundance signaling proteins in the cells or tissues. Herein we review recent advances in methods and technologies, including front-end immunoaffinity depletion, fractionation, selective enrichment of target proteins/peptides or their posttranslational modifications (PTMs), as well as advances in MS instrumentation, which have significantly enhanced the overall sensitivity of SRM assays and enabled the detection of low-abundance proteins at low to sub- ng/mL level in human blood plasma or serum. General perspectives on the potential of achieving sufficient sensitivity for detection of pg/mL level proteins in plasma are also discussed.

  11. Cellular heterogeneity mediates inherent sensitivity-specificity tradeoff in cancer targeting by synthetic circuits.

    PubMed

    Morel, Mathieu; Shtrahman, Roman; Rotter, Varda; Nissim, Lior; Bar-Ziv, Roy H

    2016-07-19

    Synthetic gene circuits are emerging as a versatile means to target cancer with enhanced specificity by combinatorial integration of multiple expression markers. Such circuits must also be tuned to be highly sensitive because escape of even a few cells might be detrimental. However, the error rates of decision-making circuits in light of cellular variability in gene expression have so far remained unexplored. Here, we measure the single-cell response function of a tunable logic AND gate acting on two promoters in heterogeneous cell populations. Our analysis reveals an inherent tradeoff between specificity and sensitivity that is controlled by the AND gate amplification gain and activation threshold. We implement a tumor-mimicking cell-culture model of cancer cells emerging in a background of normal ones, and show that molecular parameters of the synthetic circuits control specificity and sensitivity in a killing assay. This suggests that, beyond the inherent tradeoff, synthetic circuits operating in a heterogeneous environment could be optimized to efficiently target malignant state with minimal loss of specificity.

  12. RNA interference targeting Aurora-A sensitizes glioblastoma cells to temozolomide chemotherapy.

    PubMed

    Gan, Jing; Wang, Fangfang; Mu, Dezhi; Qu, Yi; Luo, Rong; Wang, Qiu

    2016-12-01

    Clinically, temozolomide (TMZ) is widely used in glioblastoma (GBM) treatment. However, the toxicity of TMZ may influence the quality of patient life. Thus, novel treatment options for sensitizing GBM cells to TMZ chemotherapy are necessary. Aurora-A is widely expressed in GBM and correlated with poor prognosis. It has been proven to be an effective target for gene therapy and chemotherapy. In the present study, short hairpin (sh)RNA targeting Aurora-A was employed to knockdown Aurora-A expression in GBM cells. Cell Counting Kit-8 assays, flow cytometry, colony formation assays, invasion assays and tube formation assays were used to determine the effects of Aurora-A knockdown when combined with TMZ treatment. A U251 subcutaneous cancer model was established to evaluate the efficacy of combined therapy. The results of the present study indicated that the proliferation, colony formation, invasion and angiogenesis of GBM cells were significantly inhibited by combined therapy when compared with TMZ treatment alone. In vivo results demonstrated that knockdown of Aurora-A significantly (P=0.0084) sensitizes GBM cells to TMZ chemotherapy. The results of the present study demonstrated that knockdown of Aurora-A in GBM cells enhances TMZ sensitivity in vitro and in vivo. Therefore, Aurora-A knockdown may be a novel treatment option for decreasing TMZ toxicity and improving patient quality of life.

  13. Targeted two-photon PDT photo-sensitizers for the treatment of subcutaneous tumors

    NASA Astrophysics Data System (ADS)

    Spangler, C. W.; Rebane, A.; Starkey, J.; Drobizhev, M.

    2009-06-01

    New porphyrin-based photo-sensitizers have been designed, synthesized and characterized that exhibit greatly enhanced intrinsic two-photon absorption. These new photo-sensitizers have been incorporated into triad formulations that also incorporate Near-infrared (NIR) imaging agents, and small-molecule targeting agents that direct the triads to cancerous tumors' over-expressed receptor sites. PDT can be initiated deep into the tissue transparency window at 780-800 nm utilizing a regeneratively amplified Ti:sapphire laser using 100-150 fs pulses of 600-800 mW. Human tumor xenografts of human breast cancer (MDA-MB-231) and both small SCLC (NCI-H69) and NSCLC (A-459) have been successfully treated using octreotate targeting of over-expressed SST2 receptors. In particular, the lung cancer xenografts can be successfully treated by irradiating from the side of the mouse opposite the implanted tumor, thereby passing through ca. 2 cm of mouse skin, tissue and organs with no discernible damage to healthy tissue while causing regression in the tumors. These results suggest a new PDT paradigm for the noninvasive treatment of subcutaneous tumors, including the possibility that the targeting moiety could be matched to individual patient genetic profiles (patient-specific therapeutics).

  14. Vinculin activators target integrins from within the cell to increase melanoma sensitivity to chemotherapy

    PubMed Central

    Nelson, Elke S.; Folkmann, Andrew W.; Henry, Michael D.; DeMali, Kris A.

    2011-01-01

    Metastatic melanoma is an aggressive skin disease for which there are no effective therapies. Emerging evidence indicates that melanomas can be sensitized to chemotherapy by increasing integrin function. Current integrin therapies work by targeting the extracellular domain, resulting in complete gains or losses of integrin function that lead to mechanism-based toxicities. An attractive alternative approach is to target proteins, such as vinculin, that associate with the integrin cytoplasmic domains and regulate its ligand binding properties. Here we report that a novel reagent, denoted vinculin activating peptide or VAP, increases integrin activity from within the cell, as measured by elevated: (1) numbers of active integrins, (2) adhesion of cells to extracellular matrix ligands, (3) numbers of cell-matrix adhesions, and (4) downstream signaling. These effects are dependent on both integrins and a key regulatory residue A50 in the vinculin head domain. We further show that VAP dramatically increases the sensitivity of melanomas to chemotherapy in clonal growth assays and in vivo mouse models of melanoma. Finally, we demonstrate that the increase in chemosensitivity results from increases in DNA damage-induced apoptosis in a p53-dependent manner. Collectively these findings demonstrate for the first time that integrin function can be manipulated from within the cell and validate integrins as a new therapeutic target for the treatment of chemoresistant melanomas. PMID:21460181

  15. Polarization of NK cell cytoskeleton upon conjugation with sensitive target cells.

    PubMed

    Carpén, O; Virtanen, I; Lehto, V P; Saksela, E

    1983-12-01

    We studied the cytoskeletal changes in natural killer (NK) cells during conjugate formation, i.e., when NK cells make contact with sensitive vs resistant target cells. F-actin and vinculin were seen to polarize at the contact sites upon conjugation with sensitive K562 cells, whereas in conjugates with resistant Raji target cells such an orientation was an infrequent finding. Myosin and two other cytoskeletal proteins, spectrin and vimentin, on the other hand, showed a random distribution in conjugating NK cells regardless of the target cell type. Hence the cytoskeletal redistribution associated with conjugation seems to be different from the receptor capping phenomenon, which is accompanied by clustering of actin, myosin, vimentin, and spectrin. On the basis of these results it seems probable that the lytic conjugate formation in NK-mediated cytotoxicity is associated with the formation of a specific type of junction that involves actin and vinculin. This cytoskeletal reorganization precedes and could be a prerequisite for the polarization of the cellular secretory apparatus and may be functionally responsible for the required cytokinetic movements.

  16. Targeting Notch, a key pathway for ovarian cancer stem cells, sensitizes tumors to platinum therapy.

    PubMed

    McAuliffe, Shannon M; Morgan, Stefanie L; Wyant, Gregory A; Tran, Lieu T; Muto, Katherine W; Chen, Yu Sarah; Chin, Kenneth T; Partridge, Justin C; Poole, Barish B; Cheng, Kuang-Hung; Daggett, John; Cullen, Kristen; Kantoff, Emily; Hasselbatt, Kathleen; Berkowitz, Julia; Muto, Michael G; Berkowitz, Ross S; Aster, Jon C; Matulonis, Ursula A; Dinulescu, Daniela M

    2012-10-23

    Chemoresistance to platinum therapy is a major obstacle that needs to be overcome in the treatment of ovarian cancer patients. The high rates and patterns of therapeutic failure seen in patients are consistent with a steady accumulation of drug-resistant cancer stem cells (CSCs). This study demonstrates that the Notch signaling pathway and Notch3 in particular are critical for the regulation of CSCs and tumor resistance to platinum. We show that Notch3 overexpression in tumor cells results in expansion of CSCs and increased platinum chemoresistance. In contrast, γ-secretase inhibitor (GSI), a Notch pathway inhibitor, depletes CSCs and increases tumor sensitivity to platinum. Similarly, a Notch3 siRNA knockdown increases the response to platinum therapy, further demonstrating that modulation of tumor chemosensitivity by GSI is Notch specific. Most importantly, the cisplatin/GSI combination is the only treatment that effectively eliminates both CSCs and the bulk of tumor cells, indicating that a dual combination targeting both populations is needed for tumor eradication. In addition, we found that the cisplatin/GSI combination therapy has a synergistic cytotoxic effect in Notch-dependent tumor cells by enhancing the DNA-damage response, G(2)/M cell-cycle arrest, and apoptosis. Based on these results, we conclude that targeting the Notch pathway could significantly increase tumor sensitivity to platinum therapy. Our study suggests important clinical applications for targeting Notch as part of novel treatment strategies upon diagnosis of ovarian cancer and at recurrence. Both platinum-resistant and platinum-sensitive relapses may benefit from such an approach as clinical data suggest that all relapses after platinum therapy are increasingly platinum resistant.

  17. Fast and sensitive detection of indels induced by precise gene targeting

    PubMed Central

    Yang, Zhang; Steentoft, Catharina; Hauge, Camilla; Hansen, Lars; Thomsen, Allan Lind; Niola, Francesco; Vester-Christensen, Malene B.; Frödin, Morten; Clausen, Henrik; Wandall, Hans H.; Bennett, Eric P.

    2015-01-01

    The nuclease-based gene editing tools are rapidly transforming capabilities for altering the genome of cells and organisms with great precision and in high throughput studies. A major limitation in application of precise gene editing lies in lack of sensitive and fast methods to detect and characterize the induced DNA changes. Precise gene editing induces double-stranded DNA breaks that are repaired by error-prone non-homologous end joining leading to introduction of insertions and deletions (indels) at the target site. These indels are often small and difficult and laborious to detect by traditional methods. Here we present a method for fast, sensitive and simple indel detection that accurately defines indel sizes down to ±1 bp. The method coined IDAA for Indel Detection by Amplicon Analysis is based on tri-primer amplicon labelling and DNA capillary electrophoresis detection, and IDAA is amenable for high throughput analysis. PMID:25753669

  18. Targeting Aurora Kinase A enhances radiation sensitivity of atypical teratoid rhabdoid tumor cells.

    PubMed

    Venkataraman, Sujatha; Alimova, Irina; Tello, Tiffany; Harris, Peter S; Knipstein, Jeffrey A; Donson, Andrew M; Foreman, Nicholas K; Liu, Arthur K; Vibhakar, Rajeev

    2012-05-01

    Atypical teratoid/rhabdoid tumors (ATRT) are rare, highly malignant, embryonal CNS tumors with a poor prognosis. Therapy relies on highly toxic chemotherapy and radiotherapy. To improve outcomes and decrease morbidity, more targeted therapy is required. Gene expression analysis revealed elevated expression of multiple kinases in ATRT tissues. Aurora Kinase A was one of the candidate kinases. The objective of this study was to evaluate the impact of Aurora Kinase A inhibition in ATRT cell lines. Our analysis revealed that inhibition of Aurora Kinase A induces cell death in ATRT cells and the small molecule inhibitor MLN 8237 sensitizes these cells to radiation. Furthermore, inhibition of Aurora Kinase A resulted in decreased activity of pro-proliferative signaling pathways. These data indicate that inhibition of Aurora Kinase A is a promising small molecule target for ATRT therapy.

  19. Potential targets by pentacyclic triterpenoids from Callicarpa farinosa against methicillin-resistant and sensitive Staphylococcus aureus.

    PubMed

    Chung, Pooi Yin; Chung, Lip Yong; Navaratnam, Parasakthi

    2014-04-01

    The evolution of antibiotic resistance in Staphylococcus aureus showed that there is no long-lasting remedy against this pathogen. The limited number of antibacterial classes and the common occurrence of cross-resistance within and between classes reinforce the urgent need to discover new compounds targeting novel cellular functions not yet targeted by currently used drugs. One of the experimental approaches used to discover novel antibacterials and their in vitro targets is natural product screening. Three known pentacyclic triterpenoids were isolated for the first time from the bark of Callicarpa farinosa Roxb. (Verbenaceae) and identified as α-amyrin [3β-hydroxy-urs-12-en-3-ol], betulinic acid [3β-hydroxy-20(29)-lupaene-28-oic acid], and betulinaldehyde [3β-hydroxy-20(29)-lupen-28-al]. These compounds exhibited antimicrobial activities against reference and clinical strains of methicillin-resistant (MRSA) and methicillin-sensitive S. aureus (MSSA), with minimum inhibitory concentration (MIC) ranging from 2 to 512 μg/mL. From the genome-wide transcriptomic analysis to elucidate the antimicrobial effects of these compounds, multiple novel cellular targets in cell division, two-component system, ABC transporters, fatty acid biosynthesis, peptidoglycan biosynthesis, aminoacyl-tRNA synthetases, ribosomes and β-lactam resistance pathways are affected, resulting in destabilization of the bacterial cell membrane, halt in protein synthesis, and inhibition of cell growth that eventually lead to cell death. The novel targets in these essential pathways could be further explored in the development of therapeutic compounds for the treatment of S. aureus infections and help mitigate resistance development due to target alterations.

  20. Phased Array Inspection of Titanium Disk Forgings Targeting no. 1/2 FBH Sensitivity

    SciTech Connect

    Roberts, R.A.; Friedl, J.

    2005-04-09

    The phased array implementation of a focused zoned ultrasonic inspection to achieve a >3dB signal-to-noise for no. 1/2 flat bottom holes (FBH) in titanium is reported. Previous work established the ultrasound focusing required to achieve the targeted sensitivity. This work reports on the design of a phased array transducer capable of maintaining the needed focus to the depths required in the forging inspection. The performance of the phased array inspection is verified by examining signal-to-noise of no. 1/2 FBHs contained in coupons cut from actual forgings.

  1. pH-sensitive oncolytic adenovirus hybrid targeting acidic tumor microenvironment and angiogenesis

    PubMed Central

    Choi, Joung-Woo; Jung, Soo-Jung; Kasala, Dayananda; Hwang, June Kyu; Hu, Jun; Bae, You Han; Yun, Chae-Ok

    2015-01-01

    Although oncolytic adenoviruses (Ads) are an attractive option for cancer gene therapy, the intravenous administration of naked Ad still encounters unfavorable host responses, non-specific interactions, and heterogeneity in targeted cancer cells. To overcome these obstacles and achieve specific targeting of the tumor microenvironment, Ad was coated with the pH-sensitive block copolymer, methoxy poly(ethylene glycol)-b-poly(l-histidine-co-l-phenylalanine) (PEGbPHF). The physicochemical properties of the generated nanocomplex, Ad/PEGbPHF, were assessed. At pH 6.4, GFP-expressing Ad/PEGbPHF induced significantly higher GFP expression than naked Ad in both coxsackie and adenovirus receptor (CAR)-positive and -negative cells. To assess the therapeutic efficacy of the Ad/PEGbPHF complex platform, an oncolytic Ad expressing VEGF promoter-targeting transcriptional repressor (KOX) was used to form complexes. At pH 6.4, KOX/PEGbPHF significantly suppressed VEGF gene expression, cancer cell migration, vessel sprouting, and cancer cell killing effect compared to naked KOX or KOX/PEGbPHF at pH 7.4, demonstrating that KOX/PEGbPHF can overcome the lack of CAR that is frequently observed in tumor tissues. The antitumor activity of KOX/PEGbPHF systemically administered to a tumor xenograft model was significantly higher than that of naked KOX. Furthermore, KOX/PEGbPHF showed lower hepatic toxicity and did not induce an innate immune response against Ad. Altogether, these results demonstrate that pH-sensitive polymer-coated Ad complex significantly increases net positive charge upon exposure to hypoxic tumor microenvironment, allowing passive targeting to the tumor tissue. It may offer superior potential for systemic therapy, due to its improved tumor selectivity, increased therapeutic efficacy, and lower toxicity compared to naked KOX. PMID:25575865

  2. pH-sensitive oncolytic adenovirus hybrid targeting acidic tumor microenvironment and angiogenesis.

    PubMed

    Choi, Joung-Woo; Jung, Soo-Jung; Kasala, Dayananda; Hwang, June Kyu; Hu, Jun; Bae, You Han; Yun, Chae-Ok

    2015-05-10

    Although oncolytic adenoviruses (Ads) are an attractive option for cancer gene therapy, the intravenous administration of naked Ad still encounters unfavorable host responses, non-specific interactions, and heterogeneity in targeted cancer cells. To overcome these obstacles and achieve specific targeting of the tumor microenvironment, Ad was coated with the pH-sensitive block copolymer, methoxy poly(ethylene glycol)-b-poly(l-histidine-co-l-phenylalanine) (PEGbPHF). The physicochemical properties of the generated nanocomplex, Ad/PEGbPHF, were assessed. At pH6.4, GFP-expressing Ad/PEGbPHF induced significantly higher GFP expression than naked Ad in both coxsackie and adenovirus receptor (CAR)-positive and -negative cells. To assess the therapeutic efficacy of the Ad/PEGbPHF complex platform, an oncolytic Ad expressing VEGF promoter-targeting transcriptional repressor (KOX) was used to form complexes. At pH6.4, KOX/PEGbPHF significantly suppressed VEGF gene expression, cancer cell migration, vessel sprouting, and cancer cell killing effect compared to naked KOX or KOX/PEGbPHF at pH7.4, demonstrating that KOX/PEGbPHF can overcome the lack of CAR that is frequently observed in tumor tissues. The antitumor activity of KOX/PEGbPHF systemically administered to a tumor xenograft model was significantly higher than that of naked KOX. Furthermore, KOX/PEGbPHF showed lower hepatic toxicity and did not induce an innate immune response against Ad. Altogether, these results demonstrate that pH-sensitive polymer-coated Ad complex significantly increases net positive charge upon exposure to hypoxic tumor microenvironment, allowing passive targeting to the tumor tissue. It may offer superior potential for systemic therapy, due to its improved tumor selectivity, increased therapeutic efficacy, and lower toxicity compared to naked KOX.

  3. Need for improved risk communication of fish consumption advisories to protect maternal and child health: influence of primary informants.

    PubMed

    LePrevost, Catherine E; Gray, Kathleen M; Hernández-Pelletier, Mercedes; Bouma, Brennan D; Arellano, Consuelo; Cope, W Gregory

    2013-04-29

    Fish consumption has established benefits, including the promotion of cardiovascular health and pre- and neonatal brain and eye development, but local freshwater fish may be a source of contaminants that are especially harmful to fetuses and young children, such as the neurotoxic and developmentally toxic methylmercury and polychlorinated biphenyls. Fish consumption advisories may be issued by state health departments to limit human exposure to these and other toxicants. This study examined the efficacy of a sign designed by the North Carolina Division of Public Health that was posted along a reservoir (Badin Lake) in central North Carolina, USA, for increasing anglers' awareness of a fish consumption advisory, with a special focus on anglers who share their catch with women and children. In this study, 109 anglers were interviewed about their awareness of fish consumption advisories in general and their knowledge of the Badin Lake fish advisory in particular. Shore anglers were significantly less likely to be aware of the term "fish consumption advisory" and of the specific advisory for Badin Lake than boat anglers. Although a significant increase in knowledge of the specific fish consumption advisory was found for the entire sample of study participants after the sign intervention, a commensurate increase in knowledge was not found for a subsample of anglers who reported sharing their catch with women and children. Study findings underscore differences in fish consumption advisory awareness among subpopulations. Specifically, the study revealed the importance of characterizing the communication needs of shore anglers and anglers who share their catch with sensitive subpopulations (e.g., women and children) for the creation of more targeted communications of fish consumption advisories.

  4. Targeting CDKs with Roscovitine Increases Sensitivity to DNA Damaging Drugs of Human Osteosarcoma Cells

    PubMed Central

    Hattinger, Claudia Maria; Fanelli, Marilù; Versteeg, Rogier; Koster, Jan; Picci, Piero

    2016-01-01

    Cyclin-dependent kinase 2 (CDK2) has been reported to be essential for cell proliferation in several human tumours and it has been suggested as an appropriate target to be considered in order to enhance the efficacy of treatment regimens based on the use of DNA damaging drugs. We evaluated the clinical impact of CDK2 overexpression on a series of 21 high-grade osteosarcoma (OS) samples profiled by using cDNA microarrays. We also assessed the in vitro efficacy of the CDKs inhibitor roscovitine in a panel of drug-sensitive and drug-resistant human OS cell lines. OS tumour samples showed an inherent overexpression of CDK2, and high expression levels at diagnosis of this kinase appeared to negatively impact on clinical outcome. CDK2 expression also proved to be relevant for in vitro OS cells growth. These findings indicated CDK2 as a promising candidate therapeutic marker for OS and therefore we assessed the efficacy of the CDKs-inhibitor roscovitine in both drug-sensitive and -resistant OS cell lines. All cell lines resulted to be responsive to roscovitine, which was also able to increase the activity of cisplatin and doxorubicin, the two most active DNA damaging drugs used in OS chemotherapy. Our results indicated that combined treatment with conventional OS chemotherapeutic drugs and roscovitine may represent a new candidate intervention approach, which may be considered to enhance tumour cell sensitivity to DNA damaging drugs. PMID:27898692

  5. Characterization of mechano-sensitive nano-containers for targeted vasodilation

    NASA Astrophysics Data System (ADS)

    Buscema, Marzia; Deyhle, Hans; Pfohl, Thomas; Hieber, Simone E.; Zumbuehl, Andreas; Müller, Bert

    2016-04-01

    Cardiovascular diseases are the worldwide number one cause of mortality. The blood flow in diseased human coronary arteries differs from the blood flow in the healthy vessels. This fact should be used for designing targeted localized delivery of vasodilators with a purely physical drug release trigger. Thus, we have proposed mechano-sensitive liposomes as mechano-sensitive containers. One has to tailor the liposome's properties, so that containers are stable under physiological conditions in health, but release their cargo near the constricted vessels at body temperature. In order to determine the shear stress threshold for release, both the morphology of the healthy and diseased human arteries and the mechanical property of the liposomes have to be known. We have shown that micro computed tomography (μCT) techniques allow visualizing the lumen of human coronary arteries and provide the basis for flow simulations to extract the wall shear stress of healthy and stenosed regions in human coronary arteries. The behavior of the mechano-sensitive liposomes is currently investigated by means of microfluidics and spatially resolved small-angle X-ray scattering. The liposomes are injected into micro-channels mimicking in vivo situation. The scattering signal from the liposomes reveals information about their size, shape, and wall thickness.

  6. A cyclic peptide targeted against PSD-95 blocks central sensitization and attenuates thermal hyperalgesia.

    PubMed

    LeBlanc, B W; Iwata, M; Mallon, A P; Rupasinghe, C N; Goebel, D J; Marshall, J; Spaller, M R; Saab, C Y

    2010-05-05

    Post-synaptic density protein PSD-95 is emerging as a valid target for modulating nociception in animal studies. Based on the key role of PSD-95 in neuronal plasticity and the maintenance of pain behavior, we predicted that CN2097, a peptide-based macrocycle of nine residues that binds to the PSD-95 Discs large, Zona occludens 1 (PDZ) domains of PSD-95, would interfere with physiologic phenomena in the spinal cord related to central sensitization. Furthermore, we tested whether spinal intrathecal injection of CN2097 attenuates thermal hyperalgesia in a rat model of sciatic neuropathy. Results demonstrate that spinal CN2097 reverses hyperexcitability of wide dynamic range (WDR) neurons in the dorsal horn of neuropathic rats and decreases their evoked responses to peripheral stimuli (brush, low caliber von Frey and pressure), whereas CN5125 ("negative control") has no effect. CN2097 also blocks C-fiber long-term potentiation (LTP) in the dorsal horn, which is linked to neuronal plasticity and central sensitization. At a molecular level, CN2097 attenuates the increase in phosphorylated p38 MAPK, a key intracellular signaling pathway in neuropathic pain. Moreover, spinal injection of CN2097 blocks thermal hyperalgesia in neuropathic rats. We conclude that CN2097 is a small molecule peptide with putative anti-nociceptive effects that modulates physiologic phenomena related to central sensitization under conditions of chronic pain.

  7. MicroRNA-595 sensitizes ovarian cancer cells to cisplatin by targeting ABCB1

    PubMed Central

    Tian, Songyu; Zhang, Mingyue; Chen, Xiuwei; Liu, Yunduo; Lou, Ge

    2016-01-01

    Ovarian cancer is among the leading cause of cancer-related deaths in females. In this study, we demonstrated that miR-595 expression was downregulated in the ovarian cancer tissues and cell lines. miR-595 expression was lower in the lymph node metastases tissues than in the primary ovarian cancer tissues and normal tissues. Furthermore, miR-595 overexpression suppressed the ovarian cancer cell proliferation, colony formation and invasion and promoted the sensitivity of ovarian cancer cell to cisplatin. We identified ABCB1 as a direct target gene of miR-595 in the ovarian cancer cell. ABCB1 expression was upregulated in the ovarian cancer tissues and cell lines. Morevoer, the expression level of ABCB1 was inversely correlated with miR-595 in the ovarian cancer tissues. In addition, overexpression of ABCB1 decreased the miR-595-overexpressing HO8910PM and SKOV-3 cell sensitivity to cisplatin. Ectopic expression of ABCB1 promoted the miR-595-overexpressing HO8910PM and SKOV-3 cell proliferation, colony formation and invasion. These data suggested that miR-595 acted a tumor suppressor role in ovarian cancer development and increased the sensitivity of ovarian cancer to cisplatin. PMID:27893429

  8. Dual-targeting peptide probe for sequence- and structure-sensitive sensing of serum albumin.

    PubMed

    Yu, Yang; Huang, Yanyan; Jin, Yulong; Zhao, Rui

    2017-04-02

    Peptide-protein interactions mediate numerous biologic processes and provide great opportunity for developing peptide probes and analytical approaches for detecting and interfering with recognition events. Molecular interactions usually take place on the heterogeneous surface of proteins, and the spatial distribution and arrangement of probes are therefore crucial for achieving high specificity and sensitivity in the bioassays. In this study, small linear peptides, homogenous peptide dimers and hetero bivalent peptides were designed for site-specific recognition of human serum albumin (HSA). Three hydrophilic regions located at different subdomains of HSA were chosen as targets for the molecular design. The binding affinity, selectivity and kinetics of the candidates were screened with surface plasmon resonance imaging (SPRi) and fluoroimmuno assays. Benefiting from the synergistic effect from the surface-targeted peptide binders and the flexible spacer, a heterogenetic dimer peptide (heter-7) with fast binding and slow dissociation behavior was identified as the optimized probe. Heter-7 specifically recognizes the target protein HSA, and effectively blocks the binding of antibody to HSA. Its inhibitory activity was estimated as 83nM. It is noteworthy that heter-7 can distinguish serum albumins from different species despite high similarities in sequence and structure of these proteins. This hetero bivalent peptide shows promise for use in serum proteomics, disease detection and drug transport, and provides an effective approach for promoting the affinity and selectivity of ligands to achieve desirable chemical and biological outcomes.

  9. MicroRNA-221 and -222 Regulate Radiation Sensitivity by Targeting the PTEN Pathway

    SciTech Connect

    Zhang Chunzhi; Kang Chunsheng; Wang Ping; Cao Yongzhen; Lv Zhonghong; Yu Shizhu; Wang Guangxiu; Zhang Anling; Jia Zhifan; Han Lei; Yang Chunying; Ishiyama, Hiromichi; Teh, Bin S.; Xu Bo; Pu Peiyu

    2011-05-01

    Purpose: MicroRNAs (miRNAs) are noncoding RNAs inhibiting expression of numerous target genes by posttranscriptional regulation. miRNA-221 and miRNA-222 (miRNA-221/-222) expression is elevated in radioresistant tumor cell lines; however, it is not known whether and how miRNAs control cellular responses to ionizing irradiation. Methods and Materials: We used bioinformatic analyses, luciferase reporter assay, and genetic knockdown and biochemical assays to characterize the regulation pathways of miRNA-221/-222 in response to radiation treatment. Results: We identified the PTEN gene as a target of miRNA-221/-222. Furthermore, we found that knocking down miRNA-221/-222 by antisense oligonucleotides upregulated PTEN expression. Upregulated PTEN expression suppressed AKT activity and increased radiation-induced apoptosis, resulting in enhancement of radiosensitivity in tumor cells. Conclusions: miRNA-221/-222 control radiation sensitivity by regulating the PTEN/AKT pathway and can be explored as novel targets for radiosensitization.

  10. A Sensitive Membrane-Targeted Biosensor for Monitoring Changes in Intracellular Chloride in Neuronal Processes

    PubMed Central

    Watts, Spencer D.; Suchland, Katherine L.; Amara, Susan G.; Ingram, Susan L.

    2012-01-01

    Background Regulation of chloride gradients is a major mechanism by which excitability is regulated in neurons. Disruption of these gradients is implicated in various diseases, including cystic fibrosis, neuropathic pain and epilepsy. Relatively few studies have addressed chloride regulation in neuronal processes because probes capable of detecting changes in small compartments over a physiological range are limited. Methodology/Principal Findings In this study, a palmitoylation sequence was added to a variant of the yellow fluorescent protein previously described as a sensitive chloride indicator (YFPQS) to target the protein to the plasma membrane (mbYFPQS) of cultured midbrain neurons. The reporter partitions to the cytoplasmic face of the cellular membranes, including the plasma membrane throughout the neurons and fluorescence is stable over 30–40 min of repeated excitation showing less than 10% decrease in mbYFPQS fluorescence compared to baseline. The mbYFPQS has similar chloride sensitivity (k50 =  41 mM) but has a shifted pKa compared to the unpalmitoylated YFPQS variant (cytYFPQS) that remains in the cytoplasm when expressed in midbrain neurons. Changes in mbYFPQS fluorescence were induced by the GABAA agonist muscimol and were similar in the soma and processes of the midbrain neurons. Amphetamine also increased mbYFPQS fluorescence in a subpopulation of cultured midbrain neurons that was reversed by the selective dopamine transporter (DAT) inhibitor, GBR12909, indicating that mbYFPQS is sensitive enough to detect endogenous DAT activity in midbrain dopamine (DA) neurons. Conclusions/Significance The mbYFPQS biosensor is a sensitive tool to study modulation of intracellular chloride levels in neuronal processes and is particularly advantageous for simultaneous whole-cell patch clamp and live-cell imaging experiments. PMID:22506078

  11. Study of the pH-sensitive mechanism of tumor-targeting liposomes.

    PubMed

    Fan, Yang; Chen, Cong; Huang, Yiheng; Zhang, Fang; Lin, Guimei

    2017-03-01

    Currently, the phosphatidylethanolamine-based, pH-sensitive, liposome drug-delivery system has been widely developed for efficient, targeted cancer therapy. However, the mechanism of pH sensitivity was unclear; it is a main obstacle in controlling the preparation of pH-sensitive liposomes (PSLs).Therefore, our research is aimed at clarifying the pH-response mechanism of the various molecules that compose liposomes. We chose the small pH-sensitive molecules oleic acid (OA), linoleic acid (LA) and cholesteryl hemisuccinate (CHEMS) and the fundamental lipids cholesterol and phosphatidylethanolamine (PE) as test molecules. The PSLs were prepared using the thin-film hydration method and characterized in detail at various pH values (pH 5.0, 6.0 and 7.4), including particle size, ζ-potential, drug encapsulation efficiency and drug loading. The surface structure was observed by transmission electron microscopy (TEM), and the electrical conductivity of the liposome dispersion was also tested. The calorimetric analysis was conducted by Nano-differential scanning calorimetry (Nano-DSC). The in vitro drug release profile showed that PSLs exhibit good pH sensitivity. At neutral pH, the particle size was approximately 150nm, and it dramatically increased at pH 5.0. The ζ-potential increased as the pH decreased. The Nano-DSC results showed that cholesterol and CHEMS can both increase the stability and phase transfer temperature of PSLs. Conductivity increased to a maximum at pH 5.0 and was rather low at pH 7.4. In conclusion, results show that the three kinds of liposomes have pH responsive release characteristics in acidic pH. The OA-PSLs have a pH sensitive point of 5. Since CHEMS has a cholesterol-like structure, it can stabilizes the phospholipid bilayer under neutral conditions as shown in the Nano-DSC data, and because it has a special steroidal rigid structure, it exhibits better pH response characteristics under acidic conditions.

  12. Sensitive Targeted Quantification of ERK Phosphorylation Dynamics and Stoichiometry in Human Cells without Affinity Enrichment

    SciTech Connect

    Shi, Tujin; Gao, Yuqian; Gaffrey, Matthew J.; Nicora, Carrie D.; Fillmore, Thomas L.; Chrisler, William B.; Gritsenko, Marina A.; Wu, Chaochao; He, Jintang; Bloodsworth, Kent J.; Zhao, Rui; Camp II, David G.; Liu, Tao; Rodland, Karin D.; Smith, Richard D.; Wiley, H. Steven; Qian, Weijun

    2014-12-17

    Mass spectrometry-based targeted quantification is a promising technology for site-specific quantification of posttranslational modifications (PTMs). However, a major constraint of most targeted MS approaches is the limited sensitivity for quantifying low-abundance PTMs, requiring the use of affinity reagents to enrich specific PTMs. Herein, we demonstrate the direct site-specific quantification of ERK phosphorylation isoforms (pT, pY, pTpY) and their relative stoichiometries using a highly sensitive targeted MS approach termed high-pressure, high-resolution separations with intelligent selection and multiplexing (PRISM). PRISM provides effective enrichment of target peptides within a given fraction from complex biological matrix with minimal sample losses, followed by selected reaction monitoring (SRM) quantification. The PRISM-SRM approach enabled direct quantification of ERK phosphorylation in human mammary epithelial cells (HMEC) from as little as 25 µg tryptic peptides from whole cell lysates. Compared to immobilized metal-ion affinity chromatography, PRISM provided >10-fold improvement in signal intensities, presumably due to the better peptide recovery of PRISM for handling small size samples. This approach was applied to quantify ERK phosphorylation dynamics in HMEC treated by different doses of EGF at both the peak activation (10 min) and steady state (2 h). At 10 min, the maximal ERK activation was observed with 0.3 ng/mL dose, whereas the maximal steady state level of ERK activation at 2 h was at 3 ng/ml dose, corresponding to 1200 and 9000 occupied receptors, respectively. At 10 min, the maximally activated pTpY isoform represented ~40% of total ERK, falling to less than 10% at 2 h. The time course and dose-response profiles of individual phosphorylated ERK isoforms indicated that singly phosphorylated pT-ERK never increases significantly, while the increase of pY-ERK paralleled that of pTpY-ERK. This data supports for a processive, rather than

  13. Sensitive Targeted Quantification of ERK Phosphorylation Dynamics and Stoichiometry in Human Cells without Affinity Enrichment

    DOE PAGES

    Shi, Tujin; Gao, Yuqian; Gaffrey, Matthew J.; ...

    2014-12-17

    Mass spectrometry-based targeted quantification is a promising technology for site-specific quantification of posttranslational modifications (PTMs). However, a major constraint of most targeted MS approaches is the limited sensitivity for quantifying low-abundance PTMs, requiring the use of affinity reagents to enrich specific PTMs. Herein, we demonstrate the direct site-specific quantification of ERK phosphorylation isoforms (pT, pY, pTpY) and their relative stoichiometries using a highly sensitive targeted MS approach termed high-pressure, high-resolution separations with intelligent selection and multiplexing (PRISM). PRISM provides effective enrichment of target peptides within a given fraction from complex biological matrix with minimal sample losses, followed by selected reactionmore » monitoring (SRM) quantification. The PRISM-SRM approach enabled direct quantification of ERK phosphorylation in human mammary epithelial cells (HMEC) from as little as 25 µg tryptic peptides from whole cell lysates. Compared to immobilized metal-ion affinity chromatography, PRISM provided >10-fold improvement in signal intensities, presumably due to the better peptide recovery of PRISM for handling small size samples. This approach was applied to quantify ERK phosphorylation dynamics in HMEC treated by different doses of EGF at both the peak activation (10 min) and steady state (2 h). At 10 min, the maximal ERK activation was observed with 0.3 ng/mL dose, whereas the maximal steady state level of ERK activation at 2 h was at 3 ng/ml dose, corresponding to 1200 and 9000 occupied receptors, respectively. At 10 min, the maximally activated pTpY isoform represented ~40% of total ERK, falling to less than 10% at 2 h. The time course and dose-response profiles of individual phosphorylated ERK isoforms indicated that singly phosphorylated pT-ERK never increases significantly, while the increase of pY-ERK paralleled that of pTpY-ERK. This data supports for a processive, rather than

  14. Advance ultra sensitive multi-layered nano plasmonic devices for label free biosensing targeting immunodiagnostics

    NASA Astrophysics Data System (ADS)

    Sharma, Divya; Dwivedi, R. P.

    2016-09-01

    The rapid advancement in technology has envisaged and drafted the use of optical bio-sensing units into label free and multiplexed bio-sensing, exploring the surface plasmon polaritons, which has turned into a gold standard on the commercial basis, but they are bulky and find difficulty in scaling up for the throughput detection. The integration of plasmonic crystals with microfluidics on the bio-sensing frontier offers a multi-level validation of results with the ease of real-time detection and imaging and holds a great promise to develop ultra-sensitive, fast, portable device for the point-of-care diagnostics. The paper describes the fast, low cost approach of designing and simulating label free biosensor using open source MEEP and other software tools targeting Immunodiagnostics.

  15. Superior sensitivity of novel molecular imaging probe: simultaneously targeting two types of endothelial injury markers

    PubMed Central

    Sun, Dawei; Nakao, Shintaro; Xie, Fang; Zandi, Souska; Schering, Alexander; Hafezi-Moghadam, Ali

    2010-01-01

    The need remains great for early diagnosis of diseases. The special structure of the eye provides a unique opportunity for noninvasive light-based imaging of fundus vasculature. To detect endothelial injury at the early and reversible stage of adhesion molecule up-regulation, we generated novel imaging agents that target two distinct types of endothelial molecules, a mediator of rolling, P-selectin, and one that mediates firm adhesion, ICAM-1. Interactions of these double-conjugated fluorescent microspheres (MSs) in retinal or choroidal microvasculature were visualized in live animals by scanning laser ophthalmoscopy. The new imaging agents showed significantly higher sensitivity for detection of endothelial injury than singly conjugated MSs (rPSGL-1- or α-ICAM-1-conjugated), both in terms of rolling (P<0.01) and firm adhesion (P<0.01). The rolling flux of α-ICAM-1-conjugated MSs did not differ in EIU animals, whereas double-conjugated MSs showed significantly higher rolling flux (P<0.01), revealing that ICAM-1 in vivo supports rolling, once MS interaction with the endothelium is initiated. Double-conjugated MSs specifically detected firmly adhering leukocytes (P<0.01), allowing in vivo quantification of immune response. Antiinflammatory treatment with dexamethasone led to reduced leukocyte accumulation (P<0.01) as well as MS interaction (P<0.01), which suggests that treatment success and resolution of inflammation is quantitatively reflected with this molecular imaging approach. This work introduces novel imaging agents for noninvasive detection of endothelial injury in vivo. Our approach may be developed further to diagnose human disease at a much earlier stage than currently possible.—Sun, D., Nakao, S., Xie, F., Zandi, S., Schering, A., Hafezi-Moghadam, A. Superior sensitivity of novel molecular imaging probe: simultaneously targeting two types of endothelial injury markers. PMID:20103715

  16. RAD50 targeting impairs DNA damage response and sensitizes human breast cancer cells to cisplatin therapy

    PubMed Central

    Flores-Pérez, Ali; Rafaelli, Lourdes E; Ramírez-Torres, Nayeli; Aréchaga-Ocampo, Elena; Frías, Sara; Sánchez, Silvia; Marchat, Laurence A; Hidalgo-Miranda, Alfredo; Quintanar-Jurado, Valeria; Rodríguez-Cuevas, Sergio; Bautista-Piña, Verónica; Carlos-Reyes, Ángeles; López-Camarillo, César

    2014-01-01

    In tumor cells the effectiveness of anti-neoplastic agents that cause cell death by induction of DNA damage is influenced by DNA repair activity. RAD50 protein plays key roles in DNA double strand breaks repair (DSBs), which is crucial to safeguard genome integrity and sustain tumor suppression. However, its role as a potential therapeutic target has not been addressed in breast cancer. Our aim in the present study was to analyze the expression of RAD50 protein in breast tumors, and evaluate the effects of RAD50-targeted inhibition on the cytotoxicity exerted by cisplatin and anthracycline and taxane-based therapies in breast cancer cells. Immunohistochemistry assays on tissue microarrays indicate that the strong staining intensity of RAD50 was reduced in 14% of breast carcinomas in comparison with normal tissues. Remarkably, RAD50 silencing by RNA interference significantly enhanced the cytotoxicity of cisplatin. Combinations of cisplatin with doxorubicin and paclitaxel drugs induced synergistic effects in early cell death of RAD50-deficient MCF-7, SKBR3, and T47D breast cancer cells. Furthermore, we found an increase in the number of DSBs, and delayed phosphorylation of histone H2AX after cisplatin treatment in RAD50-silenced cells. These cellular events were associated to a dramatical increase in the frequency of chromosomal aberrations and a decrease of cell number in metaphase. In conclusion, our data showed that RAD50 abrogation impairs DNA damage response and sensitizes breast cancer cells to cisplatin-combined therapies. We propose that the development and use of inhibitors to manipulate RAD50 levels might represent a promising strategy to sensitize breast cancer cells to DNA damaging agents. PMID:24642965

  17. Novel thermo-sensitive core-shell nanoparticles for targeted paclitaxel delivery

    NASA Astrophysics Data System (ADS)

    Li, Yuanpei; Pan, Shirong; Zhang, Wei; Du, Zhuo

    2009-02-01

    Novel thermo-sensitive nanoparticles self-assembled from poly(N,N-diethylacrylamide- co-acrylamide)-block-poly(γ-benzyl L-glutamate) were designed for targeted drug delivery in localized hyperthermia. The lower critical solution temperature (LCST) of nanoparticles was adjusted to a level between physiological body temperature (37 °C) and that used in local hyperthermia (about 43 °C). The temperature-dependent performances of the core-shell nanoparticles were systemically studied by nuclear magnetic resonance (NMR), circular dichroism (CD), fluorescence spectroscopy, dynamic light scattering (DLS), and atom force microscopy (AFM). The mean diameter of the nanoparticles increased slightly from 110 to 129 nm when paclitaxel (PTX), a poorly water-soluble anti-tumor drug, was encapsulated. A stability study in bovine serum albumin (BSA) solution indicated that the PTX loaded nanoparticles may have a long circulation time under physiological environments as the LCST was above physiological body temperature and the shell remained hydrophilic at 37 °C. The PTX release profiles showed thermo-sensitive controlled behavior. The proliferation inhibiting activity of PTX loaded nanoparticles was evaluated against Hela cells in vitro, compared with Taxol (a formulation of paclitaxel dissolved in Cremophor EL and ethanol). The cytotoxicity of PTX loaded nanoparticles increased obviously when hyperthermia was performed. The nanoparticles synthesized here could be an ideal candidate for thermal triggered anti-tumor PTX delivery system.

  18. Behavioural relevance of polarization sensitivity as a target detection mechanism in cephalopods and fishes.

    PubMed

    Pignatelli, Vincenzo; Temple, Shelby E; Chiou, Tsyr-Huei; Roberts, Nicholas W; Collin, Shaun P; Marshall, N Justin

    2011-03-12

    Aquatic habitats are rich in polarized patterns that could provide valuable information about the environment to an animal with a visual system sensitive to polarization of light. Both cephalopods and fishes have been shown to behaviourally respond to polarized light cues, suggesting that polarization sensitivity (PS) may play a role in improving target detection and/or navigation/orientation. However, while there is general agreement concerning the presence of PS in cephalopods and some fish species, its functional significance remains uncertain. Testing the role of PS in predator or prey detection seems an excellent paradigm with which to study the contribution of PS to the sensory assets of both groups, because such behaviours are critical to survival. We developed a novel experimental set-up to deliver computer-generated, controllable, polarized stimuli to free-swimming cephalopods and fishes with which we tested the behavioural relevance of PS using stimuli that evoke innate responses (such as an escape response from a looming stimulus and a pursuing behaviour of a small prey-like stimulus). We report consistent responses of cephalopods to looming stimuli presented in polarization and luminance contrast; however, none of the fishes tested responded to either the looming or the prey-like stimuli when presented in polarization contrast.

  19. Roles of NGAL and MMP-9 in the tumor microenvironment and sensitivity to targeted therapy.

    PubMed

    Candido, Saverio; Abrams, Stephen L; Steelman, Linda S; Lertpiriyapong, Kvin; Fitzgerald, Timothy L; Martelli, Alberto M; Cocco, Lucio; Montalto, Giuseppe; Cervello, Melchiorre; Polesel, Jerry; Libra, Massimo; McCubrey, James A

    2016-03-01

    Various, diverse molecules contribute to the tumor microenvironment and influence invasion and metastasis. In this review, the roles of neutrophil gelatinase-associated lipocalin (NGAL) and matrix metalloproteinase-9 (MMP-9) in the tumor microenvironment and sensitivity to therapy will be discussed. The lipocalin family of proteins has many important functions. For example when NGAL forms a complex with MMP-9 it increases its stability which is important in cancer metastasis. Small hydrophobic molecules are bound by NGAL which can alter their entry into and efflux from cells. Iron transport and storage are also influenced by NGAL activity. Regulation of iron levels is important for survival in the tumor microenvironment as well as metastasis. Innate immunity is also regulated by NGAL as it can have bacteriostatic properties. NGAL and MMP-9 expression may also affect the sensitivity of cancer cells to chemotherapy as well as targeted therapy. Thus NGAL and MMP-9 play important roles in key processes involved in metastasis as well as response to therapy. This article is part of a Special Issue entitled: Tumor Microenvironment Regulation of Cancer Cell Survival, Metastasis, Inflammation, and Immune Surveillance edited by Peter Ruvolo and Gregg L. Semenza.

  20. Behavioural relevance of polarization sensitivity as a target detection mechanism in cephalopods and fishes

    PubMed Central

    Pignatelli, Vincenzo; Temple, Shelby E.; Chiou, Tsyr-Huei; Roberts, Nicholas W.; Collin, Shaun P.; Marshall, N. Justin

    2011-01-01

    Aquatic habitats are rich in polarized patterns that could provide valuable information about the environment to an animal with a visual system sensitive to polarization of light. Both cephalopods and fishes have been shown to behaviourally respond to polarized light cues, suggesting that polarization sensitivity (PS) may play a role in improving target detection and/or navigation/orientation. However, while there is general agreement concerning the presence of PS in cephalopods and some fish species, its functional significance remains uncertain. Testing the role of PS in predator or prey detection seems an excellent paradigm with which to study the contribution of PS to the sensory assets of both groups, because such behaviours are critical to survival. We developed a novel experimental set-up to deliver computer-generated, controllable, polarized stimuli to free-swimming cephalopods and fishes with which we tested the behavioural relevance of PS using stimuli that evoke innate responses (such as an escape response from a looming stimulus and a pursuing behaviour of a small prey-like stimulus). We report consistent responses of cephalopods to looming stimuli presented in polarization and luminance contrast; however, none of the fishes tested responded to either the looming or the prey-like stimuli when presented in polarization contrast. PMID:21282177

  1. Automated divertor target design by adjoint shape sensitivity analysis and a one-shot method

    SciTech Connect

    Dekeyser, W.; Reiter, D.; Baelmans, M.

    2014-12-01

    As magnetic confinement fusion progresses towards the development of first reactor-scale devices, computational tokamak divertor design is a topic of high priority. Presently, edge plasma codes are used in a forward approach, where magnetic field and divertor geometry are manually adjusted to meet design requirements. Due to the complex edge plasma flows and large number of design variables, this method is computationally very demanding. On the other hand, efficient optimization-based design strategies have been developed in computational aerodynamics and fluid mechanics. Such an optimization approach to divertor target shape design is elaborated in the present paper. A general formulation of the design problems is given, and conditions characterizing the optimal designs are formulated. Using a continuous adjoint framework, design sensitivities can be computed at a cost of only two edge plasma simulations, independent of the number of design variables. Furthermore, by using a one-shot method the entire optimization problem can be solved at an equivalent cost of only a few forward simulations. The methodology is applied to target shape design for uniform power load, in simplified edge plasma geometry.

  2. Thermo-sensitive gels containing lorazepam microspheres for intranasal brain targeting.

    PubMed

    Jose, S; Ansa, C R; Cinu, T A; Chacko, A J; Aleykutty, N A; Ferreira, S V; Souto, E B

    2013-01-30

    Thermo-sensitive gels containing lorazepam microspheres were developed and characterized for intranasal brain targeting. Pluronics (PF-127 and PF-68) have been selected since they are thermo-reversible polymers with the property of forming a solution at low temperatures (4-5 °C), and a gel at body temperature (37 °C). This property makes them an interesting material to work with, especially in case of controlled release formulations. The present study focuses on the development of an intranasal formulation for lorazepam, as an alternative route of drug delivery to the brain. Direct transport of drugs to the brain circumventing the brain barrier, following intranasal administration, provides a unique feature and better option to target brain. The presence of mucoadhesive microspheres in the gel vehicle via nasal route can achieve a dual purpose of prolonged drug release and enhanced bioavailability. To optimise the microsphere formulation, Box Behnken design was employed by investigating the effect of three factors, polymer concentration (chitosan), emulsifier concentration (Span 80) and cross-linking agent (glutaraldehyde) on the response variable which is the mean particle size. The concentration of 21% PF-127 and 1% PF-68 were found to be promising gel vehicles. The results showed that the release rate followed a prolonged profile dispersion of the microspheres in the viscous media, in comparison to the microspheres alone. Histopathological studies proved that the optimised formulation does not produce any toxic effect on the microscopic structure of nasal mucosa.

  3. Target-induced structure-switching DNA hairpins for sensitive electrochemical monitoring of mercury (II).

    PubMed

    Zhuang, Junyang; Fu, Libing; Tang, Dianping; Xu, Mingdi; Chen, Guonan; Yang, Huanghao

    2013-01-15

    A simple, sensitive and reusable electrochemical sensor was designed for determination of mercury (II) (Hg(2+)) by coupling target-induced conformational switch of DNA hairpins with thymine-Hg(2+)-thymine (T-Hg(2+)-T) coordination chemistry. The hairpin probe consisted of a stem of 6 base pairs enclosing a 14 nucleotide (nt) loop and an additional 12 nt sticky end at the 3' end. Each hairpin was labeled with ferrocene (Fc) redox tag in the middle of the loop, which was immobilized on the electrode via self-assembly of the terminal thiol moiety at the 5' end. In the presence of target analyte, Hg(2+)-mediated base pairs induced the conformational change from the sticky end to open the hairpins, resulting in the ferrocene tags close to the electrode for the increasing redox current. The strong coordination reaction of T-Hg(2+)-T resulted in a good repeatability and intermediate precision down to 10%. The dynamic concentration range spanned from 5.0nM to 1.0μM Hg(2+) with a detection limit of 2.5nM at the 3s(blank) level. The strategy afforded exquisite selectivity for Hg(2+) against other environmentally related metal ions. Inspiringly, the developed sensor could be reused by introduction of iodide (I(-)).

  4. Point-of-care detection of extracellular vesicles: Sensitivity optimization and multiple-target detection.

    PubMed

    Oliveira-Rodríguez, Myriam; Serrano-Pertierra, Esther; García, Agustín Costa; López-Martín, Soraya; Yañez-Mo, María; Cernuda-Morollón, Eva; Blanco-López, M C

    2017-01-15

    Extracellular vesicles (EVs) are membrane-bound nanovesicles delivered by different cellular lineages under physiological and pathological conditions. Although these vesicles have shown relevance as biomarkers for a number of diseases, their isolation and detection still has several technical drawbacks, mainly related with problems of sensitivity and time-consumed. Here, we reported a rapid and multiple-targeted lateral flow immunoassay (LFIA) system for the detection of EVs isolated from human plasma. A range of different labels (colloidal gold, carbon black and magnetic nanoparticles) was compared as detection probe in LFIA, being gold nanoparticles that showed better results. Using this platform, we demonstrated that improvements may be carried out by incorporating additional capture lines with different antibodies. The device exhibited a limit of detection (LOD) of 3.4×10(6)EVs/µL when anti-CD81 and anti-CD9 were selected as capture antibodies in a multiple-targeted format, and anti-CD63 labeled with gold nanoparticles was used as detection probe. This LFIA, coupled to EVs isolation kits, could become a rapid and useful tool for the point-of-care detection of EVs, with a total analysis time of two hours.

  5. Dual tumor targeting with pH-sensitive and bioreducible polymer-complexed oncolytic adenovirus.

    PubMed

    Moon, Chang Yoon; Choi, Joung-Woo; Kasala, Dayananda; Jung, Soo-Jung; Kim, Sung Wan; Yun, Chae-Ok

    2015-02-01

    Oncolytic adenoviruses (Ads) have shown great promise in cancer gene therapy but their efficacy has been compromised by potent immunological, biochemical, and specific tumor-targeting limitations. To take full advantage of the innate cancer-specific killing potency of oncolytic Ads but also exploit the subtleties of the tumor microenvironment, we have generated a pH-sensitive and bio-reducible polymer (PPCBA)-coated oncolytic Ad. Ad-PPCBA complexes showed higher cellular uptake at pH 6.0 than pH 7.4 in both high and low coxsackie and adenovirus receptor-(CAR)-expressing cells, thereby demonstrating Ad-PPCBA's ability to target the low pH hypoxic tumor microenvironment and overcome CAR dependence for target cell uptake. Endocytic mechanism studies indicated that Ad-PPCBA internalization is mediated by macropinocytosis instead of the CAR-dependent endocytic pathway that internalizes naked Ad. VEGF-specific shRNA-expressing oncolytic Ad complexed with PPCBA (RdB/shVEGF-PPCBA) elicited much more potent suppression of U87 human brain cancer cell VEGF gene expression in vitro, and human breast cancer MCF7 cell/Matrigel plug vascularization in a mouse model, when cancer cells had been previously infected at pH 6.0 versus pH 7.4. Moreover, intratumorally and intravenously injected RdB/shVEGF-PPCBA nanocomplexes elicited significantly higher therapeutic efficacy than naked virus in U87-tumor mouse xenograft models, reducing IL-6, ALT, and AST serum levels. These data demonstrated PPCBA's biocompatibility and capability to shield the Ad surface to prevent innate immune response against Ad after both intratumoral and systemic administration. Taken together, these results demonstrate that smart, tumor-specific, oncolytic Ad-PPCBA complexes can be exploited to treat both primary and metastatic tumors.

  6. Detection of breast cancer cells using targeted magnetic nanoparticles and ultra-sensitive magnetic field sensors

    PubMed Central

    2011-01-01

    Introduction Breast cancer detection using mammography has improved clinical outcomes for many women, because mammography can detect very small (5 mm) tumors early in the course of the disease. However, mammography fails to detect 10 - 25% of tumors, and the results do not distinguish benign and malignant tumors. Reducing the false positive rate, even by a modest 10%, while improving the sensitivity, will lead to improved screening, and is a desirable and attainable goal. The emerging application of magnetic relaxometry, in particular using superconducting quantum interference device (SQUID) sensors, is fast and potentially more specific than mammography because it is designed to detect tumor-targeted iron oxide magnetic nanoparticles. Furthermore, magnetic relaxometry is theoretically more specific than MRI detection, because only target-bound nanoparticles are detected. Our group is developing antibody-conjugated magnetic nanoparticles targeted to breast cancer cells that can be detected using magnetic relaxometry. Methods To accomplish this, we identified a series of breast cancer cell lines expressing varying levels of the plasma membrane-expressed human epidermal growth factor-like receptor 2 (Her2) by flow cytometry. Anti-Her2 antibody was then conjugated to superparamagnetic iron oxide nanoparticles using the carbodiimide method. Labeled nanoparticles were incubated with breast cancer cell lines and visualized by confocal microscopy, Prussian blue histochemistry, and magnetic relaxometry. Results We demonstrated a time- and antigen concentration-dependent increase in the number of antibody-conjugated nanoparticles bound to cells. Next, anti Her2-conjugated nanoparticles injected into highly Her2-expressing tumor xenograft explants yielded a significantly higher SQUID relaxometry signal relative to unconjugated nanoparticles. Finally, labeled cells introduced into breast phantoms were measured by magnetic relaxometry, and as few as 1 million labeled cells

  7. Applied genomics: data mining reveals species-specific malaria diagnostic targets more sensitive than 18S rRNA.

    PubMed

    Demas, Allison; Oberstaller, Jenna; DeBarry, Jeremy; Lucchi, Naomi W; Srinivasamoorthy, Ganesh; Sumari, Deborah; Kabanywanyi, Abdunoor M; Villegas, Leopoldo; Escalante, Ananias A; Kachur, S Patrick; Barnwell, John W; Peterson, David S; Udhayakumar, Venkatachalam; Kissinger, Jessica C

    2011-07-01

    Accurate and rapid diagnosis of malaria infections is crucial for implementing species-appropriate treatment and saving lives. Molecular diagnostic tools are the most accurate and sensitive method of detecting Plasmodium, differentiating between Plasmodium species, and detecting subclinical infections. Despite available whole-genome sequence data for Plasmodium falciparum and P. vivax, the majority of PCR-based methods still rely on the 18S rRNA gene targets. Historically, this gene has served as the best target for diagnostic assays. However, it is limited in its ability to detect mixed infections in multiplex assay platforms without the use of nested PCR. New diagnostic targets are needed. Ideal targets will be species specific, highly sensitive, and amenable to both single-step and multiplex PCRs. We have mined the genomes of P. falciparum and P. vivax to identify species-specific, repetitive sequences that serve as new PCR targets for the detection of malaria. We show that these targets (Pvr47 and Pfr364) exist in 14 to 41 copies and are more sensitive than 18S rRNA when utilized in a single-step PCR. Parasites are routinely detected at levels of 1 to 10 parasites/μl. The reaction can be multiplexed to detect both species in a single reaction. We have examined 7 P. falciparum strains and 91 P. falciparum clinical isolates from Tanzania and 10 P. vivax strains and 96 P. vivax clinical isolates from Venezuela, and we have verified a sensitivity and specificity of ∼100% for both targets compared with a nested 18S rRNA approach. We show that bioinformatics approaches can be successfully applied to identify novel diagnostic targets and improve molecular methods for pathogen detection. These novel targets provide a powerful alternative molecular diagnostic method for the detection of P. falciparum and P. vivax in conventional or multiplex PCR platforms.

  8. SynLethDB: synthetic lethality database toward discovery of selective and sensitive anticancer drug targets.

    PubMed

    Guo, Jing; Liu, Hui; Zheng, Jie

    2016-01-04

    Synthetic lethality (SL) is a type of genetic interaction between two genes such that simultaneous perturbations of the two genes result in cell death or a dramatic decrease of cell viability, while a perturbation of either gene alone is not lethal. SL reflects the biologically endogenous difference between cancer cells and normal cells, and thus the inhibition of SL partners of genes with cancer-specific mutations could selectively kill cancer cells but spare normal cells. Therefore, SL is emerging as a promising anticancer strategy that could potentially overcome the drawbacks of traditional chemotherapies by reducing severe side effects. Researchers have developed experimental technologies and computational prediction methods to identify SL gene pairs on human and a few model species. However, there has not been a comprehensive database dedicated to collecting SL pairs and related knowledge. In this paper, we propose a comprehensive database, SynLethDB (http://histone.sce.ntu.edu.sg/SynLethDB/), which contains SL pairs collected from biochemical assays, other related databases, computational predictions and text mining results on human and four model species, i.e. mouse, fruit fly, worm and yeast. For each SL pair, a confidence score was calculated by integrating individual scores derived from different evidence sources. We also developed a statistical analysis module to estimate the druggability and sensitivity of cancer cells upon drug treatments targeting human SL partners, based on large-scale genomic data, gene expression profiles and drug sensitivity profiles on more than 1000 cancer cell lines. To help users access and mine the wealth of the data, we developed other practical functionalities, such as search and filtering, orthology search, gene set enrichment analysis. Furthermore, a user-friendly web interface has been implemented to facilitate data analysis and interpretation. With the integrated data sets and analytics functionalities, SynLethDB would

  9. Increased Insulin Sensitivity in Mice Lacking Collectrin, a Downstream Target of HNF-1α

    PubMed Central

    Malakauskas, Sandra M.; Kourany, Wissam M.; Zhang, Xiao Yin; Lu, Danhong; Stevens, Robert D.; Koves, Timothy R.; Hohmeier, Hans E.; Muoio, Deborah M.; Newgard, Christopher B.; Le, Thu H.

    2009-01-01

    Collectrin is a downstream target of the transcription factor hepatocyte nuclear factor-1α (HNF-1α), which is mutated in maturity-onset diabetes of the young subtype 3 (MODY3). Evidence from transgenic mouse models with collectrin overexpression in pancreatic islets suggests divergent roles for collectrin in influencing β-cell mass and insulin exocytosis. To clarify the function of collectrin in the pancreas, we used a mouse line with targeted deletion of the gene. We examined pancreas morphology, glucose homeostasis by ip glucose tolerance testing (IPGTT) and insulin tolerance testing (IPITT), and pancreas function by in vivo acute-phase insulin response determination and glucose-stimulated insulin secretion from isolated islets. We find no difference in either pancreas morphology or function between wild-type and collectrin-deficient animals (Tmem27−/y). However, we note that by 6 months of age, Tmem27−/y mice exhibit increased insulin sensitivity by IPITT and decreased adiposity by dual-energy x-ray absorptiometry scanning compared with wild-type. We have previously reported that Tmem27−/y mice exhibit profound aminoaciduria due to failed renal recovery. We now demonstrate that Tmem27−/y animals also display inappropriate excretion of some short-chain acylcarnitines derived from amino acid and fatty acid oxidation. We provide further evidence for compensatory up-regulation of oxidative metabolism in Tmem27−/y mice, along with enhanced protein turnover associated with preserved lean mass even out to 1.5 yr of age. Our studies suggest that collectrin-deficient mice activate a number of adaptive mechanisms to defend energy homeostasis in the setting of ongoing nutrient losses. PMID:19246514

  10. A peptide nucleic acid targeting nuclear RAD51 sensitizes multiple myeloma cells to melphalan treatment

    PubMed Central

    Alagpulinsa, David Abasiwani; Yaccoby, Shmuel; Ayyadevara, Srinivas; Shmookler Reis, Robert Joseph

    2015-01-01

    RAD51-mediated recombinational repair is elevated in multiple myeloma (MM) and predicts poor prognosis. RAD51 has been targeted to selectively sensitize and/or kill tumor cells. Here, we employed a peptide nucleic acid (PNA) to inhibit RAD51 expression in MM cells. We constructed a PNA complementary to a unique segment of the RAD51 gene promoter, spanning the transcription start site, and conjugated it to a nuclear localization signal (PKKKRKV) to enhance cellular uptake and nuclear delivery without transfection reagents. This synthetic construct, (PNArad51_nls), significantly reduced RAD51 transcripts in MM cells, and markedly reduced the number and intensity of de novo and melphalan-induced nuclear RAD51 foci, while increasing the level of melphalan-induced γH2AX foci. Melphalan alone markedly induced the expression of 5 other genes involved in homologous-recombination repair, yet suppression of RAD51 by PNArad51_nls was sufficient to synergize with melphalan, producing significant synthetic lethality of MM cells in vitro. In a SCID-rab mouse model mimicking the MM bone marrow microenvironment, treatment with PNArad51_nls ± melphalan significantly suppressed tumor growth after 2 weeks, whereas melphalan plus control PNArad4µ_nls was ineffectual. This study highlights the importance of RAD51 in myeloma growth and is the first to demonstrate that anti-RAD51 PNA can potentiate conventional MM chemotherapy. PMID:25996477

  11. Anxiety sensitivity and working memory capacity: Risk factors and targets for health behavior promotion.

    PubMed

    Otto, Michael W; Eastman, Abraham; Lo, Stephen; Hearon, Bridget A; Bickel, Warren K; Zvolensky, Michael; Smits, Jasper A J; Doan, Stacey N

    2016-11-01

    Understanding the nature and influence of specific risk profiles is increasingly important for health behavior promotion. The purpose of this article is to document the value of two factors-anxiety sensitivity (AS) and working memory capacity (WMC)-for enhancing risk for the initiation and/or maintenance of a range of negative health behaviors. AS is a distress-related risk factor that potentiates avoidance/coping motivations for negative health behaviors. Stress provides the conditions for negative somatic and affective states, and AS amplifies the aversiveness of these experiences and correspondingly hinders adaptive functioning. In contrast, low WMC is hypothesized to exert its effect by decreasing the capacity to filter out current temptations, attenuating a focus on longer-term goals and impairing the application of relevant coping skills at times of stress. In this review, we provide conceptual models for the separate roles of high AS and low WMC in negative health behaviors, review the influence of these factors on specific health behavior exemplars (eating behaviors/obesity, physical activity, smoking, alcohol use, and sleep promotion), provide preliminary evidence for their value as independent treatment targets for health-behavior promotion, and encourage specific research directions in relation to these variables.

  12. Modularity and hormone sensitivity of the Drosophila melanogaster insulin receptor/target of rapamycin interaction proteome.

    PubMed

    Glatter, Timo; Schittenhelm, Ralf B; Rinner, Oliver; Roguska, Katarzyna; Wepf, Alexander; Jünger, Martin A; Köhler, Katja; Jevtov, Irena; Choi, Hyungwon; Schmidt, Alexander; Nesvizhskii, Alexey I; Stocker, Hugo; Hafen, Ernst; Aebersold, Ruedi; Gstaiger, Matthias

    2011-11-08

    Genetic analysis in Drosophila melanogaster has been widely used to identify a system of genes that control cell growth in response to insulin and nutrients. Many of these genes encode components of the insulin receptor/target of rapamycin (InR/TOR) pathway. However, the biochemical context of this regulatory system is still poorly characterized in Drosophila. Here, we present the first quantitative study that systematically characterizes the modularity and hormone sensitivity of the interaction proteome underlying growth control by the dInR/TOR pathway. Applying quantitative affinity purification and mass spectrometry, we identified 97 high confidence protein interactions among 58 network components. In all, 22% of the detected interactions were regulated by insulin affecting membrane proximal as well as intracellular signaling complexes. Systematic functional analysis linked a subset of network components to the control of dTORC1 and dTORC2 activity. Furthermore, our data suggest the presence of three distinct dTOR kinase complexes, including the evolutionary conserved dTTT complex (Drosophila TOR, TELO2, TTI1). Subsequent genetic studies in flies suggest a role for dTTT in controlling cell growth via a dTORC1- and dTORC2-dependent mechanism.

  13. Modularity and hormone sensitivity of the Drosophila melanogaster insulin receptor/target of rapamycin interaction proteome

    PubMed Central

    Glatter, Timo; Schittenhelm, Ralf B; Rinner, Oliver; Roguska, Katarzyna; Wepf, Alexander; Jünger, Martin A; Köhler, Katja; Jevtov, Irena; Choi, Hyungwon; Schmidt, Alexander; Nesvizhskii, Alexey I; Stocker, Hugo; Hafen, Ernst; Aebersold, Ruedi; Gstaiger, Matthias

    2011-01-01

    Genetic analysis in Drosophila melanogaster has been widely used to identify a system of genes that control cell growth in response to insulin and nutrients. Many of these genes encode components of the insulin receptor/target of rapamycin (InR/TOR) pathway. However, the biochemical context of this regulatory system is still poorly characterized in Drosophila. Here, we present the first quantitative study that systematically characterizes the modularity and hormone sensitivity of the interaction proteome underlying growth control by the dInR/TOR pathway. Applying quantitative affinity purification and mass spectrometry, we identified 97 high confidence protein interactions among 58 network components. In all, 22% of the detected interactions were regulated by insulin affecting membrane proximal as well as intracellular signaling complexes. Systematic functional analysis linked a subset of network components to the control of dTORC1 and dTORC2 activity. Furthermore, our data suggest the presence of three distinct dTOR kinase complexes, including the evolutionary conserved dTTT complex (Drosophila TOR, TELO2, TTI1). Subsequent genetic studies in flies suggest a role for dTTT in controlling cell growth via a dTORC1- and dTORC2-dependent mechanism. PMID:22068330

  14. Bone-Targeted Acid-Sensitive Doxorubicin Conjugate Micelles as Potential Osteosarcoma Therapeutics

    PubMed Central

    2015-01-01

    Osteosarcoma is a malignancy of the bone that primarily affects adolescents. Current treatments retain mortality rates, which are higher than average cancer mortality rates for the adolescent age group. We designed a micellar delivery system with the aim to increase drug accumulation in the tumor and potentially reduce side effects associated with chemotherapy. The design features are the use of the hydrophilic d-aspartic acid octapeptide as both the effective targeting agent as well as the hydrophilic micelle corona. Micelle stabilization was accomplished by binding of model drug (doxorubicin) via an acid-sensitive hydrazone bond and incorporating one to four 11-aminoundecanoic acid (AUA) moieties to manipulate the hydrophobic/hydrophilic ratio. Four micelle-forming unimers have been synthesized and their self-assembly into micelles was evaluated. Size of the micelles could be modified by changing the architecture of the unimers from linear to branched. The stability of the micelles increased with increasing content of AUA moieties. Adsorption of all micelles to hydroxyapatite occurred rapidly. Doxorubicin release occurred at pH 5.5, whereas no release was detected at pH 7.4. Cytotoxicity toward human osteosarcoma Saos-2 cells correlated with drug release data. PMID:25291150

  15. Bone-targeted acid-sensitive doxorubicin conjugate micelles as potential osteosarcoma therapeutics.

    PubMed

    Low, Stewart A; Yang, Jiyuan; Kopeček, Jindřich

    2014-11-19

    Osteosarcoma is a malignancy of the bone that primarily affects adolescents. Current treatments retain mortality rates, which are higher than average cancer mortality rates for the adolescent age group. We designed a micellar delivery system with the aim to increase drug accumulation in the tumor and potentially reduce side effects associated with chemotherapy. The design features are the use of the hydrophilic D-aspartic acid octapeptide as both the effective targeting agent as well as the hydrophilic micelle corona. Micelle stabilization was accomplished by binding of model drug (doxorubicin) via an acid-sensitive hydrazone bond and incorporating one to four 11-aminoundecanoic acid (AUA) moieties to manipulate the hydrophobic/hydrophilic ratio. Four micelle-forming unimers have been synthesized and their self-assembly into micelles was evaluated. Size of the micelles could be modified by changing the architecture of the unimers from linear to branched. The stability of the micelles increased with increasing content of AUA moieties. Adsorption of all micelles to hydroxyapatite occurred rapidly. Doxorubicin release occurred at pH 5.5, whereas no release was detected at pH 7.4. Cytotoxicity toward human osteosarcoma Saos-2 cells correlated with drug release data.

  16. Targeted Deposition of Antibodies on a Multiplex CMOS Microarray and Optimization of a Sensitive Immunoassay Using Electrochemical Detection

    DTIC Science & Technology

    2010-03-19

    sandwich immunoassay used a capture Ab adsorbed to the Ppy and a reporter Ab labeled for fluorescence detection or ECD, and results from these methods of...Targeted Deposition of Antibodies on a Multiplex CMOS Microarray and Optimization of a Sensitive Immunoassay Using Electrochemical Detection John...Sensitive Immunoassay Using Electrochemical Detection 5a. CONTRACT NUMBER 5b. GRANT NUMBER 5c. PROGRAM ELEMENT NUMBER 6. AUTHOR(S) 5d. PROJECT

  17. Dual signal amplification for highly sensitive electrochemical detection of uropathogens via enzyme-based catalytic target recycling.

    PubMed

    Su, Jiao; Zhang, Haijie; Jiang, Bingying; Zheng, Huzhi; Chai, Yaqin; Yuan, Ruo; Xiang, Yun

    2011-11-15

    We report an ultrasensitive electrochemical approach for the detection of uropathogen sequence-specific DNA target. The sensing strategy involves a dual signal amplification process, which combines the signal enhancement by the enzymatic target recycling technique with the sensitivity improvement by the quantum dot (QD) layer-by-layer (LBL) assembled labels. The enzyme-based catalytic target DNA recycling process results in the use of each target DNA sequence for multiple times and leads to direct amplification of the analytical signal. Moreover, the LBL assembled QD labels can further enhance the sensitivity of the sensing system. The coupling of these two effective signal amplification strategies thus leads to low femtomolar (5fM) detection of the target DNA sequences. The proposed strategy also shows excellent discrimination between the target DNA and the single-base mismatch sequences. The advantageous intrinsic sequence-independent property of exonuclease III over other sequence-dependent enzymes makes our new dual signal amplification system a general sensing platform for monitoring ultralow level of various types of target DNA sequences.

  18. Fungicide sensitivity of US genotypes of Phytophthora infestans (Mont.) de Bary to six oomycete-targeted compounds.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Phytophthora infestans (Mont.) de Bary causes potato late blight, an important and costly disease of potato and tomato crops. The baseline sensitivity of recent clonal lineages of P. infestans was tested for six oomycete-targeted fungicides. Forty five isolates collected between 2004 and 2012 were t...

  19. Targeting Epstein-Barr virus oncoprotein LMP1-mediated glycolysis sensitizes nasopharyngeal carcinoma to radiation therapy.

    PubMed

    Xiao, L; Hu, Z-Y; Dong, X; Tan, Z; Li, W; Tang, M; Chen, L; Yang, L; Tao, Y; Jiang, Y; Li, J; Yi, B; Li, B; Fan, S; You, S; Deng, X; Hu, F; Feng, L; Bode, A M; Dong, Z; Sun, L-Q; Cao, Y

    2014-09-11

    Our goal in this work was to illustrate the Epstein-Barr virus (EBV)-modulated global biochemical profile and provide a novel metabolism-related target to improve the therapeutic regimen of nasopharyngeal carcinoma (NPC). We used a metabolomics approach to investigate EBV-modulated metabolic changes, and found that the exogenous overexpression of the EBV-encoded latent membrane protein 1 (LMP1) significantly increased glycolysis. The deregulation of several glycolytic genes, including hexokinase 2 (HK2), was determined to be responsible for the reprogramming of LMP1-mediated glucose metabolism in NPC cells. The upregulation of HK2 elevated aerobic glycolysis and facilitated proliferation by blocking apoptosis. More importantly, HK2 was positively correlated with LMP1 in NPC biopsies, and high HK2 levels were significantly associated with poor overall survival of NPC patients following radiation therapy. Knockdown of HK2 effectively enhanced the sensitivity of LMP1-overexpressing NPC cells to irradiation. Finally, c-Myc was demonstrated to be required for LMP1-induced upregulation of HK2. The LMP1-mediated attenuation of the PI3-K/Akt-GSK3beta-FBW7 signaling axis resulted in the stabilization of c-Myc. These findings indicate a close relationship between EBV and glycolysis in NPC. Notably, LMP1 is the key regulator of the reprogramming of EBV-mediated glycolysis in NPC cells. Given the importance of EBV-mediated deregulation of glycolysis, anti-glycolytic therapy might represent a worthwhile avenue of exploration in the treatment of EBV-related cancers.

  20. Herceptin conjugated PLGA-PHis-PEG pH sensitive nanoparticles for targeted and controlled drug delivery.

    PubMed

    Zhou, Zilan; Badkas, Apurva; Stevenson, Max; Lee, Joo-Youp; Leung, Yuet-Kin

    2015-06-20

    A dual functional nano-scaled drug carrier, comprising of a targeting ligand and pH sensitivity, has been made in order to increase the specificity and efficacy of the drug delivery system. The nanoparticles are made of a tri-block copolymer, poly(d,l lactide-co-glycolide) (PLGA)-b-poly(l-histidine) (PHis)-b-polyethylene glycol (PEG), via nano-precipitation. To provide the nanoparticle feature of endolysosomal escape and pH sensitivity, poly(l-histidine) was chosen as a proton sponge polymer. Herceptin, which specifically binds to HER2 antigen, was conjugated to the nanoparticles through click chemistry. The nanoparticles were characterized via dynamic light scattering (DLS) and transmission electron microscopy (TEM). Both methods showed the sizes of about 100nm with a uniform size distribution. The pH sensitivity was assessed by drug releases and size changes at different pH conditions. As pH decreased from 7.4 to 5.2, the drug release rate accelerated and the size significantly increased. During in vitro tests against human breast cancer cell lines, MCF-7 and SK-BR-3 showed significantly increased uptake for Herceptin-conjugated nanoparticles, as compared to non-targeted nanoparticles. Herceptin-conjugated pH-sensitive nanoparticles showed the highest therapeutic effect, and thus validated the efficacy of a combined approach of pH sensitivity and active targeting.

  1. Targeting the Tumour Vasculature: Exploitation of Low Oxygenation and Sensitivity to NOS Inhibition by Treatment with a Hypoxic Cytotoxin

    PubMed Central

    Baker, Jennifer H. E.; Kyle, Alastair H.; Bartels, Kirsten L.; Methot, Stephen P.; Flanagan, Erin J.; Balbirnie, Andrew; Cran, Jordan D.; Minchinton, Andrew I.

    2013-01-01

    Many cancer research efforts focus on exploiting genetic-level features that may be targeted for therapy. Tissue-level features of the tumour microenvironment also represent useful therapeutic targets. Here we investigate the presence of low oxygen tension and sensitivity to NOS inhibition of tumour vasculature as potential tumour-specific features that may be targeted by hypoxic cytotoxins, a class of therapeutics currently under investigation. We have previously demonstrated that tirapazamine (TPZ) mediates central vascular dysfunction in tumours. TPZ is a hypoxic cytotoxin that is also a competitive inhibitor of NOS. Here we further investigated the vascular-targeting activity of TPZ by combining it with NOS inhibitor L-NNA, or with low oxygen content gas breathing. Tumours were analyzed via multiplex immunohistochemical staining that revealed irreversible loss of perfusion and enhanced tumour cell death when TPZ was combined with either low oxygen or a NOS inhibitor. Tumour growth rate was reduced by TPZ + NOS inhibition, and tumours previously resistant to TPZ-mediated vascular dysfunction were sensitized by low oxygen breathing. Additional mapping analysis suggests that tumours with reduced vascular-associated stroma may have greater sensitivity to these effects. These results indicate that poorly oxygenated tumour vessels, also being abnormally organized and with inadequate smooth muscle, may be successfully targeted for significant anti-cancer effects by inhibition of NOS and hypoxia-activated prodrug toxicity. This strategy illustrates a novel use of hypoxia-activated cytotoxic prodrugs as vascular targeting agents, and also represents a novel mechanism for targeting tumour vessels. PMID:24204680

  2. An experimental correlation study between field-target overlap and sensitivity of surface plasmon resonance biosensors based on sandwiched immunoassays

    NASA Astrophysics Data System (ADS)

    Ryu, Yeonsoo; Moon, Seyoung; Oh, Youngjin; Kim, Yonghwi; Kim, Donghyun

    2012-10-01

    In this report, we have studied the effectiveness of field-target overlap to evaluate detection sensitivity of surface plasmon resonance (SPR) biosensors. The investigation used theoretical analysis based on the transfer matrix method, which was experimentally confirmed by thin film-based detection in sandwich and reverse sandwich immunoglobulin G (IgG) assays. Both theoretical and experimental results show that strong correlation exists between the overlap and the sensitivity with the coefficient of correlation higher than 95% in all the cases that we have considered. We have also confirmed the correlation in diffraction grating-based SPR measurement of IgG/anti-IgG interactions. The correlation elucidates the mechanism behind the far-field detection sensitivity of SPR biosensors and can lead to the enhancement of SPR biosensing with molecular scale sensitivity.

  3. Locally targeted delivery of a micron-size radiation therapy source using temperature-sensitive hydrogel

    PubMed Central

    Kim, Y; Seol, DR; Mohapatra, S; Sunderland, JJ; Schultz, MK; Domann, FE; Lim, TH

    2014-01-01

    Purpose To propose a novel radiotherapy (RT) delivery modality—locally targeted delivery of micro-size level RT sources using temperature-sensitive hydroGEL (RT-GEL) as an injectable vehicle. Methods and Materials Hydrogel is a water-like liquid at room temperature but gels at body temperature. Two FDA-approved polymers were synthesized. Indium-111 (In-111) was used as a radioactive RT-GEL source. The release characteristics of In-111 from polymerized RT-GEL were evaluated. The injectability and efficacy of RT-GEL delivery to human breast tumor were tested using animal models with control datasets of RT-Saline injection. As proof-of-concept studies, a total of 6 nude mice were tested by injecting 4 million tumor cells into their upper backs after a week of acclimatization. Three mice were injected with RT-GEL and three with RT-Saline. A SPECT and a CT scan were performed on each mouse at 0, 24, and 48 hours after injection. The efficacy of RT-GEL over the control datasets was determined by measuring kidney In-111 accumulation (mean nCi/cc), representing the distant diffusion of In-111. Results RT-GEL was successfully injected to the tumor using 30-gauge needles. No difficulties due to polymerization of hydrogel during injection and intratumoral pressure were observed during RT-GEL injection. No back flow occurred for either RT-GEL or RT-Saline. The residual tumor activities of In-111 were 49% (44%) at 24 hours (48 hour) for RT-GEL and 29% (22%) for RT-Saline. SPECT-CT fused images of RT-Saline show considerable kidney accumulation of In-111 (2,886%, 261% and 262% of RT-GEL at the 0, 24 and 48 hour marks, respectively). Conclusion RT-GEL was successfully injected and showed much higher residual tumor activity 170% (200%) than that of RT-Saline 24 hour (48 hour) after injection with a minimal accumulation of In-111 to the kidneys. The preliminary data of RT-GEL as a delivery modality of a radiation source to a local tumor is promising. PMID:24495593

  4. Locally Targeted Delivery of a Micron-Size Radiation Therapy Source Using Temperature-Sensitive Hydrogel

    SciTech Connect

    Kim, Yusung; Seol, Dong Rim; Mohapatra, Sucheta; Sunderland, John J.; Schultz, Michael K.; Domann, Frederick E.; Lim, Tae-Hong

    2014-04-01

    Purpose: To propose a novel radiation therapy (RT) delivery modality: locally targeted delivery of micron-size RT sources by using temperature-sensitive hydrogel (RT-GEL) as an injectable vehicle. Methods and Materials: Hydrogel is a water-like liquid at room temperature but gels at body temperature. Two US Food and Drug Administration-approved polymers were synthesized. Indium-111 (In-111) was used as the radioactive RT-GEL source. The release characteristics of In-111 from polymerized RT-GEL were evaluated. The injectability and efficacy of RT-GEL delivery to human breast tumor were tested using animal models with control datasets of RT-saline injection. As proof-of-concept studies, a total of 6 nude mice were tested by injecting 4 million tumor cells into their upper backs after a week of acclimatization. Three mice were injected with RT-GEL and 3 with RT-saline. Single-photon emission computed tomography (SPECT) and CT scans were performed on each mouse at 0, 24, and 48 h after injection. The efficacy of RT-GEL was determined by comparison with that of the control datasets by measuring kidney In-111 accumulation (mean nCi/cc), representing the distant diffusion of In-111. Results: RT-GEL was successfully injected into the tumor by using a 30-gauge needle. No difficulties due to polymerization of hydrogel during injection and intratumoral pressure were observed during RT-GEL injection. No back flow occurred for either RT-GEL or RT-saline. The residual tumor activities of In-111 were 49% at 24 h (44% at 48 h, respectively) for RT-GEL and 29% (22%, respectively) for RT-saline. Fused SPECT-CT images of RT-saline showed considerable kidney accumulation of In-111 (2886%, 261%, and 262% of RT-GEL at 0, 24, and 48 h, respectively). Conclusions: RT-GEL was successfully injected and showed much higher residual tumor activity: 170% (200%, respectively), than that of RT-saline at 24 h (48 h, respectively) after injection with a minimal accumulation of In-111 to the

  5. Targeted inhibition of HDAC8 increases the doxorubicin sensitivity of neuroblastoma cells via up regulation of miR-137.

    PubMed

    Zhao, Gang; Wang, Guoliang; Bai, Hongmin; Li, Tiandong; Gong, Fanghe; Yang, Huan; Wen, Jinchong; Wang, Weimin

    2017-02-20

    Histone deacetylases (HDACs) have been suggested to be potential therapeutic targets for cancer treatment. Recent studies revealed that HDAC8 expression was associated with poor prognostic markers and poor overall survival rate of neuroblastoma (NB). Our present study revealed that among the four members of class I HDACs, HDAC8 is significantly over expressed in NB cells as compared with the normal fibroblast 3T3 cells or primary normal human astrocytes (NHA) cells. Targeted inhibition of HDAC8 by its specific siRNA (si-HDAC8) can inhibit the in vitro growth of NB cells. Furthermore, si-HDAC8 significantly increases the sensitivity of NB cells to doxorubicin (Dox). Silencing of HDAC8 can increase the expression of miR-137, which has been suggested to mediate the Dox sensitivity of NB cells. Knockdown of miR-137 can attenuate si-HDAC8 enhanced Dox sensitivity. Further, si-HDAC8 can also inhibit the expression of multi-drug resistance gene 1 (MDR1). While knockdown of miR-137 can attenuate si-HDAC8 induced down regulation of MDR1. Collectively, our data revealed that targeted inhibition of HDAC8 can suppress the growth of NB cells and increase Dox sensitivity via up regulation of miR-137 and suppression of MDR1. Therefor, combination of HDAC8 inhibitor will be helpful to elevate the treatment outcome of NB patients.

  6. RT-PCR using glycoprotein target is more sensitive for the detection of Ebola virus in clinical samples.

    PubMed

    Yang, Mingjuan; Ke, Yuehua; Zhang, Wenyi; Liu, Chao; Yang, Ruifu; Chen, Zeliang

    2017-03-01

    The recent largest ever Ebola virus disease (EVD) outbreak in West Africa has been of worldwide concern, causing huge economic losses and constituting serious threat to the local residents and health care workers. Rapid detection of Ebola virus (EBOV) using RT-PCR has been suggested to be of great value in stopping the outbreak, because it is highly sensitive and specific and can return results within hours. In this study, 210 clinical samples, including 109 blood and 101 nasopharyngeal swab samples were used to compare the performance of glycoprotein (GP) and nucleoprotein (NP) gene targets for the detection of EBOV. The analytical sensitivity of both assays were 10 molecules/μL. For clinical samples, the sensitivity of the assay targeting GP gene is higher than that of NP gene (respectively 98% and 94%) and the specificities for both targets were 100%. In addition, the positive samples in the RT-PCR assay targeting GP showed lower cycle threshold values and higher virus loads than NP gene.

  7. Smaller Fixation Target Size Is Associated with More Stable Fixation and Less Variance in Threshold Sensitivity

    PubMed Central

    Okano, Kana; Koshiji, Risako; Funaki, Wakana; Shoji, Nobuyuki

    2016-01-01

    The aims of this randomized observational case control study were to quantify fixation behavior during standard automated perimetry (SAP) with different fixation targets and to evaluate the relationship between fixation behavior and threshold variability at each test point in healthy young participants experienced with perimetry. SAP was performed on the right eyes of 29 participants using the Octopus 900 perimeter, program 32, dynamic strategy. The fixation targets of Point, Cross, and Ring were used for SAP. Fixation behavior was recorded using a wearable eye-tracking glass. All participants underwent SAP twice with each fixation target in a random fashion. Fixation behavior was quantified by calculating the bivariate contour ellipse area (BCEA) and the frequency of deviation from the fixation target. The BCEAs (deg2) of Point, Cross, and Ring targets were 1.11, 1.46, and 2.02, respectively. In all cases, BCEA increased significantly with increasing fixation target size (p < 0.05). The logarithmic value of BCEA demonstrated the same tendency (p < 0.05). A positive correlation was identified between fixation behavior and threshold variability for the Point and Cross targets (ρ = 0.413–0.534, p < 0.05). Fixation behavior increased with increasing fixation target size. Moreover, a larger fixation behavior tended to be associated with a higher threshold variability. A small fixation target is recommended during the visual field test. PMID:27829030

  8. Multifunctional pH-sensitive superparamagnetic iron-oxide nanocomposites for targeted drug delivery and MR imaging.

    PubMed

    Zhu, Lijuan; Wang, Dali; Wei, Xuan; Zhu, Xinyuan; Li, Jianqi; Tu, Chunlai; Su, Yue; Wu, Jieli; Zhu, Bangshang; Yan, Deyue

    2013-08-10

    A multifunctional pH-sensitive superparamagnetic iron-oxide (SPIO) nanocomposite system was developed for simultaneous tumor magnetic resonance imaging (MRI) and therapy. Small-size SPIO nanoparticles were chemically bonded with antitumor drug doxorubicin (DOX) and biocompatible poly(ethylene glycol) (PEG) through pH-sensitive acylhydrazone linkages, resulting in the formation of SPIO nanocomposites with magnetic targeting and pH-sensitive properties. These DOX-conjugated SPIO nanocomposites exhibited not only good stability in aqueous solution but also high saturation magnetizations. Under an acidic environment, the DOX was quickly released from the SPIO nanocomposites due to the cleavage of pH-sensitive acylhydrazone linkages. With the help of magnetic field, the DOX-conjugated SPIO nanocomposites showed high cellular uptake, indicating their magnetic targeting property. Comparing to free DOX, the DOX-conjugated SPIO nanocomposites showed better antitumor effect under magnetic field. At the same time, the relaxivity value of these SPIO nanocomposites was higher than 146s(-1)mM(-1) Fe, leading to ~4 times enhancement compared to that of free SPIO nanoparticles. As a negative contrast agent, these SPIO nanocomposites illustrated high resolution in MRI diagnosis of tumor-bearing mice. All of these results confirm that these pH-sensitive SPIO nanocomposites are promising hybrid materials for synergistic MRI diagnosis and tumor therapy.

  9. Application of exonuclease III-aided target recycling in flow cytometry: DNA detection sensitivity enhanced by orders of magnitude.

    PubMed

    Lu, Jie; Paulsen, Ian T; Jin, Dayong

    2013-09-03

    DNA-functionalized microspheres in conjugation with flow cytometry detection are widely used for high-throughput nucleic acid assays. Although such assays are rapid and capable of simultaneous analysis of multiple nucleic acid analytes in a single test, the intrinsic limitation in sensitivity remains challenging. Here we report a simple, highly sensitive, and reproducible method based on Exonuclease III-aided target recycling technique applied for DNA quantification in flow cytometry. By loading a high density of Cy5-labeled probe DNA on microspheres (15 μm), we achieved hitherto unreported DNA detection limit of 3.2 pM in flow cytometry bead assay, enhancing the sensitivity by a factor of over 56.8 compared to the conventional direct hybridization bead assay. Furthermore, we evaluated multiplexing capability by simultaneous detections of two target DNAs with FAM and Cy5 reporter conjugated probes. Therefore, the novel Exonuclease III-amplified flow cytometry bead assay has great potential for the rapid, sensitive, and accurate detection and quantification of nucleic acids in clinical diagnosis and biomedical research.

  10. THZ1 targeting CDK7 suppresses STAT transcriptional activity and sensitizes T-cell lymphomas to BCL2 inhibitors

    PubMed Central

    Cayrol, Florencia; Praditsuktavorn, Pannee; Fernando, Tharu M.; Kwiatkowski, Nicholas; Marullo, Rosella; Calvo-Vidal, M. Nieves; Phillip, Jude; Pera, Benet; Yang, Shao Ning; Takpradit, Kaipol; Roman, Lidia; Gaudiano, Marcello; Crescenzo, Ramona; Ruan, Jia; Inghirami, Giorgio; Zhang, Tinghu; Cremaschi, Graciela; Gray, Nathanael S.; Cerchietti, Leandro

    2017-01-01

    Peripheral T-cell lymphomas (PTCL) are aggressive diseases with poor response to chemotherapy and dismal survival. Identification of effective strategies to target PTCL biology represents an urgent need. Here we report that PTCL are sensitive to transcription-targeting drugs, and, in particular, to THZ1, a covalent inhibitor of cyclin-dependent kinase 7 (CDK7). The STAT-signalling pathway is highly vulnerable to THZ1 even in PTCL cells that carry the activating STAT3 mutation Y640F. In mutant cells, CDK7 inhibition decreases STAT3 chromatin binding and expression of highly transcribed target genes like MYC, PIM1, MCL1, CD30, IL2RA, CDC25A and IL4R. In surviving cells, THZ1 decreases the expression of STAT-regulated anti-apoptotic BH3 family members MCL1 and BCL-XL sensitizing PTCL cells to BH3 mimetic drugs. Accordingly, the combination of THZ1 and the BH3 mimetic obatoclax improves lymphoma growth control in a primary PTCL ex vivo culture and in two STAT3-mutant PTCL xenografts, delineating a potential targeted agent-based therapeutic option for these patients. PMID:28134252

  11. Integrating 2-D position sensitive X-ray detectors with low-density alkali halide storage targets

    NASA Astrophysics Data System (ADS)

    Haubold, H.-G.; Hoheisel, W.; Hiller, P.

    1986-05-01

    For the use in scattering experiments with synchrotron radiation, integrating position sensitive X-ray detectors are discussed. These detectors store the photon number equivalent charge (PNEC) in low-density alkali halide targets. Performance tests are given for a detector which uses a Gd 2O 2S fluorescence screen for X-ray detection and the low-density KCl storage target of a television SEC vidicon tube for photon integration. Rather than directly by X-rays, this target is charged by 6 keV electrons from the image intensifier section of the vidicon. Its excellent storage capability allows measurements of extremely high-contrast, high-flux X-ray patterns with the same accuracy as achieved with any single photon detection system if the discussed readout techniques are applied.

  12. Lysophosphatidylcholine hydrolases of human erythrocytes, lymphocytes, and brain: Sensitive targets of conserved specificity for organophosphorus delayed neurotoxicants

    SciTech Connect

    Vose, Sarah C.; Holland, Nina T.; Eskenazi, Brenda; Casida, John E.

    2007-10-01

    Brain neuropathy target esterase (NTE), associated with organophosphorus (OP)-induced delayed neuropathy, has the same OP inhibitor sensitivity and specificity profiles assayed in the classical way (paraoxon-resistant, mipafox-sensitive hydrolysis of phenyl valerate) or with lysophosphatidylcholine (LysoPC) as the substrate. Extending our earlier observation with mice, we now examine human erythrocyte, lymphocyte, and brain LysoPC hydrolases as possible sensitive targets for OP delayed neurotoxicants and insecticides. Inhibitor profiling of human erythrocytes and lymphocytes gave the surprising result of essentially the same pattern as with brain. Human erythrocyte LysoPC hydrolases are highly sensitive to OP delayed neurotoxicants, with in vitro IC{sub 50} values of 0.13-85 nM for longer alkyl analogs, and poorly sensitive to the current OP insecticides. In agricultural workers, erythrocyte LysoPC hydrolyzing activities are similar for newborn children and their mothers and do not vary with paraoxonase status but have high intersample variation that limits their use as a biomarker. Mouse erythrocyte LysoPC hydrolase activity is also of low sensitivity in vitro and in vivo to the OP insecticides whereas the delayed neurotoxicant ethyl n-octylphosphonyl fluoride inhibits activity in vivo at 1-3 mg/kg. Overall, inhibition of blood LysoPC hydrolases is as good as inhibition of brain NTE as a predictor of OP inducers of delayed neuropathy. NTE and lysophospholipases (LysoPLAs) both hydrolyze LysoPC, yet they are in distinct enzyme families with no sequence homology and very different catalytic sites. The relative contributions of NTE and LysoPLAs to LysoPC hydrolysis and clearance from erythrocytes, lymphocytes, and brain remain to be defined.

  13. Advisory Council Member.

    ERIC Educational Resources Information Center

    Illinois State Board of Vocational Education and Rehabilitation, Springfield. Div. of Vocational and Technical Education.

    The pamphlet is directed toward the new member of a career education advisory council. It explains why advisory councils are needed and why an individual should join one. An advisory council is defined as a group of persons selected to collectively advise regarding career education efforts within the community, whose members are predominantly from…

  14. pH-sensitive siRNA Nanovector for Targeted Gene Silencing and Cytotoxic Effect in Cancer Cells

    PubMed Central

    Mok, Hyejung; Veiseh, Omid; Fang, Chen; Kievit, Forrest M.; Wang, Freddy Y.; Park, James O.; Zhang, Miqin

    2010-01-01

    A small interfering RNA (siRNA) nanovector with dual targeting specificity and dual therapeutic effect is developed for targeted cancer imaging and therapy. The nanovector is comprised of an iron oxide magnetic nanoparticle core coated with three different functional molecules: polyethyleneimine (PEI), siRNA, and chlorotoxin (CTX). The primary amine group of PEI is blocked with citraconic anhydride that is removable at acidic conditions, not only to increase its biocompatibility at physiological conditions but also to elicit a pH-sensitive cytotoxic effect in the acidic tumor microenvironment. The PEI is covalently immobilized on the nanovector via a disulfide linkage that is cleavable after cellular internalization of the nanovector. CTX as a tumor-specific targeting ligand and siRNA as a therapeutic payload are conjugated on the nanovector via a flexible and hydrophilic PEG linker for targeted gene silencing in cancer cells. With a size of ~ 60 nm, the nanovector exhibits long-term stability and good magnetic property for magnetic resonance imaging. The multifunctional nanovector exhibits both significant cytotoxic and gene silencing effects at acidic pH conditions for C6 glioma cells, but not at physiological pH conditions. Our results suggest that this nanovector system could be safely used as a potential therapeutic agent for targeted treatment of glioma as well as other cancers. PMID:20722417

  15. Combinatorial high-throughput experimental and bioinformatic approach identifies molecular pathways linked with the sensitivity to anticancer target drugs

    PubMed Central

    Venkova, Larisa; Aliper, Alexander; Suntsova, Maria; Kholodenko, Roman; Shepelin, Denis; Borisov, Nicolas; Malakhova, Galina; Vasilov, Raif; Roumiantsev, Sergey; Zhavoronkov, Alex; Buzdin, Anton

    2015-01-01

    Effective choice of anticancer drugs is important problem of modern medicine. We developed a method termed OncoFinder for the analysis of new type of biomarkers reflecting activation of intracellular signaling and metabolic molecular pathways. These biomarkers may be linked with the sensitivity to anticancer drugs. In this study, we compared the experimental data obtained in our laboratory and in the Genomics of Drug Sensitivity in Cancer (GDS) project for testing response to anticancer drugs and transcriptomes of various human cell lines. The microarray-based profiling of transcriptomes was performed for the cell lines before the addition of drugs to the medium, and experimental growth inhibition curves were built for each drug, featuring characteristic IC50 values. We assayed here four target drugs - Pazopanib, Sorafenib, Sunitinib and Temsirolimus, and 238 different cell lines, of which 11 were profiled in our laboratory and 227 - in GDS project. Using the OncoFinder-processed transcriptomic data on ∼600 molecular pathways, we identified pathways showing significant correlation between pathway activation strength (PAS) and IC50 values for these drugs. Correlations reflect relationships between response to drug and pathway activation features. We intersected the results and found molecular pathways significantly correlated in both our assay and GDS project. For most of these pathways, we generated molecular models of their interaction with known molecular target(s) of the respective drugs. For the first time, our study uncovered mechanisms underlying cancer cell response to drugs at the high-throughput molecular interactomic level. PMID:26317900

  16. Function and Mechanism of Axonal Targeting of Voltage-sensitive Potassium Channels

    PubMed Central

    Gu, Chen; Barry, Joshua

    2011-01-01

    Precise localization of various ion channels into proper subcellular compartments is crucial for neuronal excitability and synaptic transmission. Axonal K+ channels that are activated by depolarization of the membrane potential participate in the repolarizing phase of the action potential, and hence regulate action potential firing patterns, which encode output signals. Moreover, some of these channels can directly control neurotransmitter release at axonal terminals by constraining local membrane excitability and limiting Ca2+ influx. K+ channels differ not only in biophysical and pharmacological properties, but in expression and subcellular distribution as well. Importantly, proper targeting of channel proteins is a prerequisite for electrical and chemical functions of axons. In this review, we first highlight recent studies that demonstrate different roles of axonal K+ channels in the local regulation of axonal excitability. Next, we focus on research progress in identifying axonal targeting motifs and machinery of several different types of K+ channels present in axons. Regulation of K+ channel targeting and activity may underlie a novel form of neuronal plasticity. This research field can contribute to generating novel therapeutic strategies through manipulating neuronal excitability in treating neurological diseases, such as multiple sclerosis, neuropathic pain, and Alzheimer’s disease. PMID:21530607

  17. NRF2 Regulates HER2 and HER3 Signaling Pathway to Modulate Sensitivity to Targeted Immunotherapies.

    PubMed

    Khalil, Hilal S; Langdon, Simon P; Kankia, Ibrahim H; Bown, James; Deeni, Yusuf Y

    2016-01-01

    NF-E2 related factor-2 (NRF2) is an essential transcription factor for multiple genes encoding antioxidants and detoxification enzymes. NRF2 is implicated in promoting cancer therapeutic resistance by its detoxification function and crosstalk with proproliferative pathways. However, the exact mechanism of this intricate connectivity between NRF2 and growth factor induced proliferative pathway remains elusive. Here, we have demonstrated that pharmacological activation of NRF2 by tert-butylhydroquinone (tBHQ) upregulates the HER family receptors, HER2 and HER3 expression, elevates pAKT levels, and enhances the proliferation of ovarian cancer cells. Preactivation of NRF2 also attenuates the combined growth inhibitory effects of HER2 targeting monoclonal antibodies, Pertuzumab and Trastuzumab. Further, tBHQ caused transcriptional induction of HER2 and HER3, while SiRNA-mediated knockdown of NRF2 prevented this and further caused transcriptional repression and enhanced cytotoxicity of the HER2 inhibitors. Hence, NRF2 regulates both HER2 and HER3 receptors to influence cellular responses to HER2 targeting monoclonal antibodies. This deciphered crosstalk mechanism reinforces the role of NRF2 in drug resistance and as a relevant anticancer target.

  18. KEAP1 loss modulates sensitivity to kinase targeted therapy in lung cancer

    PubMed Central

    Krall, Elsa B; Wang, Belinda; Munoz, Diana M; Ilic, Nina; Raghavan, Srivatsan; Niederst, Matthew J; Yu, Kristine; Ruddy, David A; Aguirre, Andrew J; Kim, Jong Wook; Redig, Amanda J; Gainor, Justin F; Williams, Juliet A; Asara, John M; Doench, John G; Janne, Pasi A; Shaw, Alice T; McDonald III, Robert E; Engelman, Jeffrey A; Stegmeier, Frank; Schlabach, Michael R; Hahn, William C

    2017-01-01

    Inhibitors that target the receptor tyrosine kinase (RTK)/Ras/mitogen-activated protein kinase (MAPK) pathway have led to clinical responses in lung and other cancers, but some patients fail to respond and in those that do resistance inevitably occurs (Balak et al., 2006; Kosaka et al., 2006; Rudin et al., 2013; Wagle et al., 2011). To understand intrinsic and acquired resistance to inhibition of MAPK signaling, we performed CRISPR-Cas9 gene deletion screens in the setting of BRAF, MEK, EGFR, and ALK inhibition. Loss of KEAP1, a negative regulator of NFE2L2/NRF2, modulated the response to BRAF, MEK, EGFR, and ALK inhibition in BRAF-, NRAS-, KRAS-, EGFR-, and ALK-mutant lung cancer cells. Treatment with inhibitors targeting the RTK/MAPK pathway increased reactive oxygen species (ROS) in cells with intact KEAP1, and loss of KEAP1 abrogated this increase. In addition, loss of KEAP1 altered cell metabolism to allow cells to proliferate in the absence of MAPK signaling. These observations suggest that alterations in the KEAP1/NRF2 pathway may promote survival in the presence of multiple inhibitors targeting the RTK/Ras/MAPK pathway. DOI: http://dx.doi.org/10.7554/eLife.18970.001 PMID:28145866

  19. Sensitivity of postplanning target and OAR coverage estimates to dosimetric margin distribution sampling parameters

    SciTech Connect

    Xu Huijun; Gordon, J. James; Siebers, Jeffrey V.

    2011-02-15

    Purpose: A dosimetric margin (DM) is the margin in a specified direction between a structure and a specified isodose surface, corresponding to a prescription or tolerance dose. The dosimetric margin distribution (DMD) is the distribution of DMs over all directions. Given a geometric uncertainty model, representing inter- or intrafraction setup uncertainties or internal organ motion, the DMD can be used to calculate coverage Q, which is the probability that a realized target or organ-at-risk (OAR) dose metric D{sub v} exceeds the corresponding prescription or tolerance dose. Postplanning coverage evaluation quantifies the percentage of uncertainties for which target and OAR structures meet their intended dose constraints. The goal of the present work is to evaluate coverage probabilities for 28 prostate treatment plans to determine DMD sampling parameters that ensure adequate accuracy for postplanning coverage estimates. Methods: Normally distributed interfraction setup uncertainties were applied to 28 plans for localized prostate cancer, with prescribed dose of 79.2 Gy and 10 mm clinical target volume to planning target volume (CTV-to-PTV) margins. Using angular or isotropic sampling techniques, dosimetric margins were determined for the CTV, bladder and rectum, assuming shift invariance of the dose distribution. For angular sampling, DMDs were sampled at fixed angular intervals {omega} (e.g., {omega}=1 deg., 2 deg., 5 deg., 10 deg., 20 deg.). Isotropic samples were uniformly distributed on the unit sphere resulting in variable angular increments, but were calculated for the same number of sampling directions as angular DMDs, and accordingly characterized by the effective angular increment {omega}{sub eff}. In each direction, the DM was calculated by moving the structure in radial steps of size {delta}(=0.1,0.2,0.5,1 mm) until the specified isodose was crossed. Coverage estimation accuracy {Delta}Q was quantified as a function of the sampling parameters {omega} or

  20. Bcl-xL inhibition by molecular-targeting drugs sensitizes human pancreatic cancer cells to TRAIL.

    PubMed

    Hari, Yoko; Harashima, Nanae; Tajima, Yoshitsugu; Harada, Mamoru

    2015-12-08

    Tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) induces apoptosis in various types of cancer cells without damaging normal cells. However, in terms of pancreatic cancer, not all cancer cells are sensitive to TRAIL. In this study, we examined a panel of human pancreatic cancer cell lines for TRAIL sensitivity and investigated the effects of Bcl-2 family inhibitors on their response to TRAIL. Both ABT-263 and ABT-737 inhibited the function of Bcl-2, Bcl-xL, and Bcl-w. Of the nine pancreatic cancer cell lines tested, six showed no or low sensitivity to TRAIL, which correlated with protein expression of Bcl-xL. ABT-263 significantly sensitized four cell lines (AsPC-1, Panc-1, CFPAC-1, and Panc10.05) to TRAIL, with reduced cell viability and increased apoptosis. Knockdown of Bcl-xL, but not Bcl-2, by siRNA transfection increased the sensitivity of AsPC-1 and Panc-1 cells to TRAIL. ABT-263 treatment had no effect on protein expression of Bcl-2, Bcl-xL, or c-FLIPs. In Panc-1 cells, ABT-263 increased the surface expression of death receptor (DR) 5; the NF-κB pathway, but not endoplasmic reticulum stress, participated in the increase. In xenograft mouse models, the combination of TRAIL and ATB-737 suppressed the in vivo tumor growth of AsPC-1 and Panc-1 cells. These results indicate that Bcl-xL is responsible for TRAIL resistance in human pancreatic cancer cells, and that Bcl-2 family inhibitors could represent promising reagents to sensitize human pancreatic cancers in DR-targeting therapy.

  1. Effects of targeted phosphorylation site mutations in the DNA-PKcs phosphorylation domain on low and high LET radiation sensitivity.

    PubMed

    Cartwright, Ian M; Bell, Justin J; Maeda, Junko; Genet, Matthew D; Romero, Ashley; Fujii, Yoshihiro; Fujimori, Akira; Kitamuta, Hisashi; Kamada, Tadashi; Chen, David J; Kato, Takamitsu A

    2015-04-01

    The present study investigated the effect of targeted mutations in the DNA-dependent protein kinase catalytic subunit and phosphorylation domains on the survival of cells in response to different qualities of ionizing radiation. Mutated Chinese hamster ovary V3 cells were exposed to 500 MeV/nucleon initial energy and 200 keV/μm monoenergetic Fe ions; 290 MeV/nucleon initial energy and average 50 keV/μm spread-out Bragg peak C ions; 70 MeV/nucleon initial energy and 1 keV/μm monoenergetic protons; and 0.663 MeV initial energy and 0.3 keV/μm Cs(137) γ radiation. The results demonstrated that sensitivity to high linear energy transfer radiation is increased when both S2056 and T2609 clusters each contain a point mutation or multiple mutations are present in either cluster, whereas the phosphoinositide 3 kinase cluster only requires a single mutation to induce the sensitized phenotype of V3 cells. Additionally, the present study demonstrated that sensitivity to DNA cross-linking damage by cisplatin only requires a single mutation in one of the three clusters and that additional point mutations do not increase cell sensitivity.

  2. Temperature-sensitive polymer-coated magnetic nanoparticles as a potential drug delivery system for targeted therapy of thyroid cancer.

    PubMed

    Koppolu, Bhanuprasanth; Bhavsar, Zarna; Wadajkar, Aniket S; Nattama, Sivaniarvindpriya; Rahimi, Maham; Nwariaku, Fiemu; Nguyen, Kytai T

    2012-12-01

    The objective of this work was to develop and investigate temperature-sensitive poly(N-isopropylacrylamide-acrylamide-allylamine)-coated iron oxide magnetic nanoparticles (TPMNPs) as possible targeted drug carriers for treatments of advanced thyroid cancer (ATC). These nanoparticles were prepared by free radical polymerization of monomers on the surface of silane-coupled iron oxide nanoparticles. In vitro studies demonstrated that TPMNPs were cytocompatible and effectively taken up by cancer cells in a dose-dependent manner. An external magnetic field significantly increased nanoparticle uptake, especially when cells were exposed to physiological flow conditions. Drug loading and release studies using doxorubicin confirmed the temperature-responsive release of drugs from nanoparticles. In addition, doxorubicin-loaded nanoparticles significantly killed ATC cells when compared to free doxorubicin. The in vitro results indicate that TPMNPs have potential as targeted and controlled drug carriers for thyroid cancer treatment.

  3. Sensitizing mucoepidermoid carcinomas to chemotherapy by targeted disruption of cancer stem cells

    PubMed Central

    Martins, Manoela D.; Warner, Kristy A.; Silva, Alan R. S.; Vargas, Pablo A.; Nunes, Fabio D.; Squarize, Cristiane H.; Nör, Jacques E.; Castilho, Rogerio M.

    2016-01-01

    Mucoepidermoid carcinoma (MEC) is the most common malignancy of salivary glands. The response of MEC to chemotherapy is unpredictable, and recent advances in cancer biology suggest the involvement of cancer stem cells (CSCs) in tumor progression and chemoresistance and radioresistance phenotype. We found that histone acetyltransferase inhibitors (HDACi) were capable of disrupting CSCs in MEC. Furthermore, administration of HDACi prior to Cisplatin (two-hit approach) disrupts CSCs and sensitizes tumor cells to Cisplatin. Our findings corroborate to emerging evidence that CSCs play a key role in tumor resistance to chemotherapy, and highlights a pharmacological two-hit approach that disrupts tumor resistance to conventional therapy. PMID:27285758

  4. KEAP1 loss modulates sensitivity to kinase targeted therapy in lung cancer. | Office of Cancer Genomics

    Cancer.gov

    Inhibitors that target the receptor tyrosine kinase (RTK)/Ras/mitogen-activated protein kinase (MAPK) pathway have led to clinical responses in lung and other cancers, but some patients fail to respond and in those that do resistance inevitably occurs (Balak et al., 2006; Kosaka et al., 2006; Rudin et al., 2013; Wagle et al., 2011). To understand intrinsic and acquired resistance to inhibition of MAPK signaling, we performed CRISPR-Cas9 gene deletion screens in the setting of BRAF, MEK, EGFR, and ALK inhibition.

  5. Targeting miR-21 enhances the sensitivity of human colon cancer HT-29 cells to chemoradiotherapy in vitro

    SciTech Connect

    Deng, Jun; Lei, Wan; Fu, Jian-Chun; Zhang, Ling; Li, Jun-He; Xiong, Jian-Ping

    2014-01-17

    Highlight: •MiR-21 plays a significant role in 5-FU resistance. •This role might be attributed to targeting of hMSH2 as well as TP and DPD via miR-21 targeted hMSH2. •Indirectly targeted TP and DPD to influence 5-FU chemotherapy sensitivity. -- Abstract: 5-Fluorouracil (5-FU) is a classic chemotherapeutic drug that has been widely used for colorectal cancer treatment, but colorectal cancer cells are often resistant to primary or acquired 5-FU therapy. Several studies have shown that miR-21 is significantly elevated in colorectal cancer. This suggests that this miRNA might play a role in this resistance. In this study, we investigated this possibility and the possible mechanism underlying this role. We showed that forced expression of miR-21 significantly inhibited apoptosis, enhanced cell proliferation, invasion, and colony formation ability, promoted G1/S cell cycle transition and increased the resistance of tumor cells to 5-FU and X radiation in HT-29 colon cancer cells. Furthermore, knockdown of miR-21 reversed these effects on HT-29 cells and increased the sensitivity of HT-29/5-FU to 5-FU chemotherapy. Finally, we showed that miR-21 targeted the human mutS homolog2 (hMSH2), and indirectly regulated the expression of thymidine phosphorylase (TP) and dihydropyrimidine dehydrogenase (DPD). These results demonstrate that miR-21 may play an important role in the 5-FU resistance of colon cancer cells.

  6. Targeting of apoptotic pathways by SMAC or BH3 mimetics distinctly sensitizes paclitaxel-resistant triple negative breast cancer cells.

    PubMed

    Panayotopoulou, Effrosini G; Müller, Anna-Katharina; Börries, Melanie; Busch, Hauke; Hu, Guohong; Lev, Sima

    2017-02-06

    Standard chemotherapy is the only systemic treatment for triple-negative breast cancer (TNBC), and despite the good initial response, resistance remains a major therapeutic obstacle. Here, we employed a High-Throughput Screen to identify targeted therapies that overcome chemoresistance in TNBC. We applied short-term paclitaxel treatment and screened 320 small-molecule inhibitors of known targets to identify drugs that preferentially and efficiently target paclitaxel-treated TNBC cells. Among these compounds the SMAC mimetics (BV6, Birinapant) and BH3-mimetics (ABT-737/263) were recognized as potent targeted therapy for multiple paclitaxel-residual TNBC cell lines. However, acquired paclitaxel resistance through repeated paclitaxel pulses result in desensitization to BV6, but not to ABT-263, suggesting that short- and long-term paclitaxel resistance are mediated by distinct mechanisms. Gene expression profiling of paclitaxel-residual, -resistant and naïve MDA-MB-231 cells demonstrated that paclitaxel-residual, as opposed to -resistant cells, were characterized by an apoptotic signature, with downregulation of anti-apoptotic genes (BCL2, BIRC5), induction of apoptosis inducers (IL24, PDCD4), and enrichment of TNFα/NF-κB pathway, including upregulation of TNFSF15, coupled with cell-cycle arrest. BIRC5 and FOXM1 downregulation and IL24 induction was also evident in breast cancer patient datasets following taxane treatment. Exposure of naïve or paclitaxel-resistant cells to supernatants of paclitaxel-residual cells sensitized them to BV6, and treatment with TNFα enhanced BV6 potency, suggesting that sensitization to BV6 is mediated, at least partially, by secreted factor(s). Our results suggest that administration of SMAC or BH3 mimetics following short-term paclitaxel treatment could be an effective therapeutic strategy for TNBC, while only BH3-mimetics could effectively overcome long-term paclitaxel resistance.

  7. Asteroid Target Selection and Orbital Manipulation Sensitivity for Deflection Demonstration Missions

    NASA Astrophysics Data System (ADS)

    Sanchez, J. P.

    2015-06-01

    In recent years, space agencies have begun to seriously consider launching demonstration missions to test some of the asteroid orbital deflection technologies and methods that have been studied and discussed in the scientific literature. Consequently, several mission studies have already been carried out. This paper attempts to gain new insights into the target selection process by analyzing the orbital evolution of a large set of notional accessible asteroids that cover all types of Near Earth Object families. The evolution of their unperturbed orbits and their anthropogenically modified trajectories was compared, and a measure of the resilience of a given orbit to anthropogenic manipulation was taken (i.e., orbital innocuity). The results show that pruning criteria such as considering only Amor objects (i.e., non-Earth-crossers) reduce unnecessarily the population of potential suitable targets and that within large regions of Earth-crossing orbital space asteroids can be found that are both accessible and safe to manipulate from the standpoint of the Earth impact risk.

  8. Transferrin receptor-targeted pH-sensitive micellar system for diminution of drug resistance and targetable delivery in multidrug-resistant breast cancer

    PubMed Central

    Gao, Wei; Ye, Guihua; Duan, Xiaochuan; Yang, Xiaoying; Yang, Victor C

    2017-01-01

    The emergence of drug resistance is partially associated with overproduction of transferrin receptor (TfR). To overcome multidrug resistance (MDR) and achieve tumor target delivery, we designed a novel biodegradable pH-sensitive micellar system modified with HAIYPRH, a TfR ligand (7pep). First, the polymers poly(l-histidine)-coupled polyethylene glycol-2000 (PHIS-PEG2000) and 7pep-modified 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-polyethylene glycol-2000 (7pep-DSPE-PEG2000) were synthesized, and the mixed micelles were prepared by blending of PHIS-PEG2000 and 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-polyethylene glycol-2000 (DSPE-PEG2000) or 7pep-DSPE-PEG2000 (7-pep HD micelles). The micelles exhibited good size uniformity, high encapsulation efficiency, and a low critical micelle concentration. By changing the polymer ratio in the micellar formulation, the pH response range was specially tailored to pH ~6.0. When loaded with antitumor drug doxorubicin (DOX), the micelle showed an acid pH-triggering drug release profile. The cellular uptake and cytotoxicity study demonstrated that 7-pep HD micelles could significantly enhance the intracellular level and antitumor efficacy of DOX in multidrug-resistant cells (MCF-7/Adr), which attributed to the synergistic effect of poly(l-histidine)-triggered endolysosom escape and TfR-mediated endocytosis. Most importantly, the in vivo imaging study confirmed the target-ability of 7-pep HD micelles to MDR tumor. These findings indicated that 7-pep HD micelles would be a promising drug delivery system in the treatment of drug-resistant tumors. PMID:28223798

  9. Imatinib-sensitive tyrosine kinases regulate mycobacterial pathogenesis and represent therapeutic targets against tuberculosis.

    PubMed

    Napier, Ruth J; Rafi, Wasiulla; Cheruvu, Mani; Powell, Kimberly R; Zaunbrecher, M Analise; Bornmann, William; Salgame, Padmini; Shinnick, Thomas M; Kalman, Daniel

    2011-11-17

    The lengthy course of treatment with currently used antimycobacterial drugs and the resulting emergence of drug-resistant strains have intensified the need for alternative therapies against Mycobacterium tuberculosis (Mtb), the etiologic agent of tuberculosis. We show that Mtb and Mycobacterium marinum use ABL and related tyrosine kinases for entry and intracellular survival in macrophages. In mice, the ABL family tyrosine kinase inhibitor, imatinib (Gleevec), when administered prophylactically or therapeutically, reduced both the number of granulomatous lesions and bacterial load in infected organs and was also effective against a rifampicin-resistant strain. Further, when coadministered with current first-line drugs, rifampicin or rifabutin, imatinib acted synergistically. These data implicate host tyrosine kinases in entry and intracellular survival of mycobacteria and suggest that imatinib may have therapeutic efficacy against Mtb. Because imatinib targets host, it is less likely to engender resistance compared to conventional antibiotics and may decrease the development of resistance against coadministered drugs.

  10. Increased sensitivity to ionizing radiation by targeting the homologous recombination pathway in glioma initiating cells.

    PubMed

    Lim, Yi Chieh; Roberts, Tara L; Day, Bryan W; Stringer, Brett W; Kozlov, Sergei; Fazry, Shazrul; Bruce, Zara C; Ensbey, Kathleen S; Walker, David G; Boyd, Andrew W; Lavin, Martin F

    2014-12-01

    Glioblastoma is deemed the most malignant form of brain tumour, particularly due to its resistance to conventional treatments. A small surviving group of aberrant stem cells termed glioma initiation cells (GICs) that escape surgical debulking are suggested to be the cause of this resistance. Relatively quiescent in nature, GICs are capable of driving tumour recurrence and undergo lineage differentiation. Most importantly, these GICs are resistant to radiotherapy, suggesting that radioresistance contribute to their survival. In a previous study, we demonstrated that GICs had a restricted double strand break (DSB) repair pathway involving predominantly homologous recombination (HR) associated with a lack of functional G1/S checkpoint arrest. This unusual behaviour led to less efficient non-homologous end joining (NHEJ) repair and overall slower DNA DSB repair kinetics. To determine whether specific targeting of the HR pathway with small molecule inhibitors could increase GIC radiosensitivity, we used the Ataxia-telangiectasia mutated inhibitor (ATMi) to ablate HR and the DNA-dependent protein kinase inhibitor (DNA-PKi) to inhibit NHEJ. Pre-treatment with ATMi prior to ionizing radiation (IR) exposure prevented HR-mediated DNA DSB repair as measured by Rad51 foci accumulation. Increased cell death in vitro and improved in vivo animal survival could be observed with combined ATMi and IR treatment. Conversely, DNA-PKi treatment had minimal impact on GICs ability to resolve DNA DSB after IR with only partial reduction in cell survival, confirming the major role of HR. These results provide a mechanistic insight into the predominant form of DNA DSB repair in GICs, which when targeted may be a potential translational approach to increase patient survival.

  11. pH-sensitive K(+) channel TREK-1 is a novel target in pancreatic cancer.

    PubMed

    Sauter, Daniel R P; Sørensen, Christiane E; Rapedius, Markus; Brüggemann, Andrea; Novak, Ivana

    2016-10-01

    Pancreatic ductal adenocarcinoma (PDAC) is one of the most lethal cancers and new therapeutic targets are urgently needed. One of the hallmarks of cancer is changed pH-homeostasis and potentially pH-sensors may play an important role in cancer cell behavior. Two-pore potassium channels (K2P) are pH-regulated channels that conduct a background K(+) current, which is involved in setting the plasma membrane potential (Vm). Some members of the K2P superfamily were reported as crucial players in driving tumor progression. The aim of this study was to investigate pH-regulated K(+) currents in PDAC cells and determine possible effects on their pathological phenotype. Using a planar high-throughput patch-clamp system (SyncroPatch 384PE) we identified a pH-regulated K(+) current in the PDAC cell line BxPC-3. The current was inhibited by extracellular acidification and intracellular alkalization. Exposure to a set of different K(+) channel inhibitors, and the TREK-1 (K2P2.1)-specific activator BL1249, TREK-1 was identified as the main component of pH-regulated current. A voltage-sensor dye (VF2.1.Cl) was used to monitor effects of pH and BL1249 on Vm in more physiological conditions and TREK-1-mediated current was found as critical player in setting Vm. We assessed a possible role of TREK-1 in PDAC progression using cell proliferation and migration assays and observed similar trends with attenuated proliferation/migration rates in acidic (pH<7.0) and alkaline (pH>7.4) conditions. Notably, BL1249 inhibited both PDAC cell proliferation and migration indicating that hyperpolarization of Vm attenuates cancer cell behavior. TREK-1 may therefore be a promising novel target for PDAC therapy.

  12. MicroRNA-31 sensitizes human breast cells to apoptosis by direct targeting of protein kinase C epsilon (PKCepsilon).

    PubMed

    Körner, Cindy; Keklikoglou, Ioanna; Bender, Christian; Wörner, Angelika; Münstermann, Ewald; Wiemann, Stefan

    2013-03-22

    MicroRNAs post-transcriptionally regulate gene expression and thereby contribute to the modulation of numerous complex and disease-relevant cellular phenotypes, including cell proliferation, cell motility, apoptosis, and stress response. In breast cancer cell systems, miR-31 has been shown to inhibit cell migration, invasion, and metastasis. Here, we link enhanced expression of miR-31 to the inhibition of the oncogenic NF-κB pathway, thus supporting the tumor-suppressive function of this microRNA. We identified protein kinase C epsilon (PKCε encoded by the PRKCE gene) as a novel direct target of miR-31 and show that down-regulation of PKCε results in impaired NF-κB signaling, enhanced apoptosis, and increased sensitivity of MCF10A breast epithelial and MDA-MB-231 triple-negative breast cancer cells toward ionizing radiation as well as treatment with chemotherapeutics. Mechanistically, we attribute this sensitization to anti-cancer treatments to the PRKCE-mediated down-regulation of the anti-apoptotic factor BCL2. In clinical breast cancer samples, high BCL2 expression was associated with poor prognosis. Furthermore, we found an inverse correlation between miR-31 and BCL2 expression, highlighting the functional relevance of the indirect down-regulation of BCL2 via direct targeting of PRKCE by miR-31.

  13. miR-145 sensitizes breast cancer to doxorubicin by targeting multidrug resistance-associated protein-1

    PubMed Central

    Gao, Man; Miao, Lingling; Liu, Mingxia; Li, Chenggang; Yu, Cunzhi; Yan, Hong; Yin, Yongxiang; Wang, Yizheng; Qi, Xinming; Ren, Jin

    2016-01-01

    Multidrug resistance-associated protein 1 (MRP1) is an important efflux transporter and overexpression of MRP1 usually leads to chemoresistance in breast cancer. Here, we found MRP1 overexpressed in human breast cancer tissues and breast cancer cell lines (compared with normal breast tissues and cell line, respectively). And MRP1 level increased in doxorubicin resistant MCF-7 cells compared with parental MCF-7 cells. Increasing evidences suggest microRNAs (miRNAs) influence chemotherapy response. We found miR-145 level decreased in human breast cancer tissues, breast cancer cell lines and doxorubicin resistant MCF-7 cells, and inversely correlated with MRP1 expression level. In the process of constructing MCF-7 doxorubicin resistant cell line, escalating doxorubicin markedly decreased miR-145 level, following by increased MRP1 level. Further study showed, miR-145 suppressed MRP1 expression by directly targeting MRP1 3′-untranslated regions. Overexpression of miR-145 sensitized breast cancer cells to doxorubicin in vitro and enhanced to doxorubicin chemotherapy in vivo through inducing intracellular doxorubicin accumulation via inhibiting MRP1. Taken together, our study revealed miR-145 sensitizes breast cancer to doxorubicin by targeting MRP1 and indicated the potential application in developing MRP1 inhibitor. PMID:27487127

  14. A geranyl acetophenone targeting cysteinyl leukotriene synthesis prevents allergic airway inflammation in ovalbumin-sensitized mice

    SciTech Connect

    Ismail, Norazren; Jambari, Nuzul Nurahya; Zareen, Seema; Akhtar, Mohamad Nadeem; Shaari, Khozirah; Zamri-Saad, Mohamad; Tham, Chau Ling; Sulaiman, Mohd Roslan; Lajis, Nordin Hj; Israf, Daud Ahmad

    2012-03-01

    Asthma is associated with increased pulmonary inflammation and airway hyperresponsiveness. The current use of corticosteroids in the management of asthma has recently raised issues regarding safety and lack of responsiveness in 5–10% of asthmatic individuals. The aim of the present study was to investigate the therapeutic effect of a non-steroidal small molecule that has cysteinyl leukotriene (cysLT) inhibitory activity, upon attenuation of allergic lung inflammation in an acute murine model. Mice were sensitized with ovalbumin (OVA) and treated with several intraperitoneal doses (100, 20, 2 and 0.2 mg/kg) of 2,4,6,-trihydroxy-3-geranylacetophenone (tHGA). Bronchoalveolar lavage was performed, blood and lung samples were obtained and respiratory function was measured. OVA sensitization increased pulmonary inflammation and pulmonary allergic inflammation was significantly reduced at doses of 100, 20 and 2 mg/kg with no effect at the lowest dose of 0.2 mg/kg. The beneficial effects in the lung were associated with reduced eosinophilic infiltration and reduced secretion of Th2 cytokines and cysLTs. Peripheral blood reduction of total IgE was also a prominent feature. Treatment with tHGA significantly attenuated altered airway hyperresponsiveness as measured by the enhanced pause (Penh) response to incremental doses of methacholine. These data demonstrate that tHGA, a synthetic non-steroidal small molecule, can prevent acute allergic inflammation. This proof of concept opens further avenues of research and development of tHGA as an additional option to the current armamentarium of anti-asthma therapeutics. -- Highlights: ► Safer and effective anti-asthmatic drugs are in great demand. ► tHGA is a new 5-LO/cysLT inhibitor that inhibits allergic asthma in mice. ► tHGA is a natural compound that can be synthesized. ► Doses as low as 2 mg/kg alleviate lung pathology in experimental asthma. ► tHGA is a potential drug lead for the treatment of allergic asthma.

  15. Targeting DNA repair with aphidicolin sensitizes primary chronic lymphocytic leukemia cells to purine analogs

    PubMed Central

    Starczewska, Eliza; Beyaert, Maxime; Michaux, Lucienne; Vekemans, Marie-Christiane; Saussoy, Pascale; Bol, Vanesa; Echarri, Ainhoa Arana; Smal, Caroline; Van Den Neste, Eric; Bontemps, Françoise

    2016-01-01

    Purine analogs are among the most effective chemotherapeutic drugs for the treatment of chronic lymphocytic leukemia (CLL). However, chemoresistance and toxicity limit their clinical use. Here, we report that the DNA polymerase inhibitor aphidicolin, which displayed negligible cytotoxicity as a single agent in primary CLL cells, markedly synergizes with fludarabine and cladribine via enhanced apoptosis. Importantly, synergy was recorded regardless of CLL prognostic markers. At the molecular level, aphidicolin enhanced purine analog-induced phosphorylation of p53 and accumulation of γH2AX, consistent with increase in DNA damage. In addition, aphidicolin delayed γH2AX disappearance that arises after removal of purine analogs, suggesting that aphidicolin causes an increase in DNA damage by impeding DNA damage repair. Similarly, aphidicolin inhibited UV-induced DNA repair known to occur primarily through the nucleotide excision repair (NER) pathway. Finally, we showed that fludarabine induced nuclear import of XPA, an indispensable factor for NER, and that XPA silencing sensitized cell lines to undergo apoptosis in response to fludarabine. Together, our data indicate that aphidicolin potentiates the cytotoxicity of purine analogs by inhibiting a DNA repair pathway that involves DNA polymerases, most likely NER, and provide a rationale for manipulating it to therapeutic advantage. PMID:27223263

  16. Sensitization of acute lymphoblastic leukemia cells for LCL161-induced cell death by targeting redox homeostasis.

    PubMed

    Haß, Christina; Belz, Katharina; Schoeneberger, Hannah; Fulda, Simone

    2016-04-01

    Disturbed redox homeostasis with both elevated reactive oxygen species (ROS) levels and antioxidant defense mechanisms has been reported in acute lymphoblastic leukemia (ALL). We therefore hypothesized that inhibition of pathways responsible for ROS detoxification renders ALL cells more susceptible for cell death. Here, we report that pharmacological inhibitors of key pathways for the elimination of ROS, i.e. Erastin, buthionine sulfoximine (BSO) and Auranofin, sensitize ALL cells for cell death upon treatment with the Smac mimetic LCL161 that antagonizes Inhibitor of Apoptosis (IAP) proteins. Erastin, BSO or Auranofin significantly increase LCL161-induced cell death and also act in concert with LCL161 to profoundly suppress long-term clonogenic survival in several ALL cell lines. Erastin or BSO cooperates with LCL161 to stimulate ROS production and lipid peroxidation prior to cell death. ROS production and lipid peroxidation are required for this cotreatment-induced cell death, since ROS scavengers or pharmacological inhibition of lipid peroxidation provides significant protection against cell death. These results emphasize that inhibition of antioxidant defense mechanisms can serve as a potent approach to prime ALL cells for LCL161-induced cell death.

  17. End-binding proteins sensitize microtubules to the action of microtubule-targeting agents.

    PubMed

    Mohan, Renu; Katrukha, Eugene A; Doodhi, Harinath; Smal, Ihor; Meijering, Erik; Kapitein, Lukas C; Steinmetz, Michel O; Akhmanova, Anna

    2013-05-28

    Microtubule-targeting agents (MTAs) are widely used for treatment of cancer and other diseases, and a detailed understanding of the mechanism of their action is important for the development of improved microtubule-directed therapies. Although there is a large body of data on the interactions of different MTAs with purified tubulin and microtubules, much less is known about how the effects of MTAs are modulated by microtubule-associated proteins. Among the regulatory factors with a potential to have a strong impact on MTA activity are the microtubule plus end-tracking proteins, which control multiple aspects of microtubule dynamic instability. Here, we reconstituted microtubule dynamics in vitro to investigate the influence of end-binding proteins (EBs), the core components of the microtubule plus end-tracking protein machinery, on the effects that MTAs exert on microtubule plus-end growth. We found that EBs promote microtubule catastrophe induction in the presence of all MTAs tested. Analysis of microtubule growth times supported the view that catastrophes are microtubule age dependent. This analysis indicated that MTAs affect microtubule aging in multiple ways: destabilizing MTAs, such as colchicine and vinblastine, accelerate aging in an EB-dependent manner, whereas stabilizing MTAs, such as paclitaxel and peloruside A, induce not only catastrophes but also rescues and can reverse the aging process.

  18. miR-320 enhances the sensitivity of human colon cancer cells to chemoradiotherapy in vitro by targeting FOXM1

    SciTech Connect

    Wan, Lu-Ying; Deng, Jun; Xiang, Xiao-Jun; Zhang, Ling; Yu, Feng; Chen, Jun; Sun, Zhe; Feng, Miao; Xiong, Jian-Ping

    2015-02-06

    Highlights: • miR-320 plays a significant role in chemoresistance. • This role might be attribute to targeting FOXM1. • The Wnt/β-catenin pathway also involves in this chemotherapy sensitivity. - Abstract: miR-320 expression level is found to be down-regulated in human colon cancer. To date, however, its underlying mechanisms in the chemo-resistance remain largely unknown. In this study, we demonstrated that ectopic expression of miR-320 led to inhibit HCT-116 cell proliferation, invasion and hypersensitivity to 5-Fu and Oxaliplatin. Also, knockdown of miR-320 reversed these effects in HT-29 cells. Furthermore, we identified an oncogene, FOXM1, as a direct target of miR-320. In addition, miR-320 could inactive the activity of Wnt/β-catenin pathway. Finally, we found that miR-320 and FOXM1 protein had a negative correlation in colon cancer tissues and adjacent normal tissues. These findings implied that miR-320–FOXM1 axis may overcome chemo-resistance of colon cancer cells and provide a new therapeutic target for the treatment of colon cancer.

  19. Carbocysteine restores steroid sensitivity by targeting histone deacetylase 2 in a thiol/GSH-dependent manner.

    PubMed

    Song, Yun; Lu, Hao-Zhong; Xu, Jian-Rong; Wang, Xiao-Lin; Zhou, Wei; Hou, Li-Na; Zhu, Liang; Yu, Zhi-Hua; Chen, Hong-Zhuan; Cui, Yong-Yao

    2015-01-01

    Steroid insensitivity is commonly observed in patients with chronic obstructive pulmonary disease. Here, we report the effects and mechanisms of carbocysteine (S-CMC), a mucolytic agent, in cellular and animal models of oxidative stress-mediated steroid insensitivity. The following results were obtained: oxidative stress induced higher levels of interleukin-8 (IL-8) and tumor necrosis factor alpha (TNF-α), which are insensitive to dexamethasone (DEX). The failure of DEX was improved by the addition of S-CMC by increasing histone deacetylase 2 (HDAC2) expression/activity. S-CMC also counteracted the oxidative stress-induced increase in reactive oxygen species (ROS) levels and decreases in glutathione (GSH) levels and superoxide dismutase (SOD) activity. Moreover, oxidative stress-induced events were decreased by the thiol-reducing agent dithiothreitol (DTT), enhanced by the thiol-oxidizing agent diamide, and the ability of DEX was strengthened by DTT. In addition, the oxidative stress-induced decrease in HDAC2 activity was counteracted by S-CMC by increasing thiol/GSH levels, which exhibited a direct interaction with HDAC2. S-CMC treatment increased HDAC2 recruitment and suppressed H4 acetylation of the IL-8 promoter, and this effect was further ablated by addition of buthionine sulfoximine, a specific inhibitor of GSH synthesis. Our results indicate that S-CMC restored steroid sensitivity by increasing HDAC2 expression/activity in a thiol/GSH-dependent manner and suggest that S-CMC may be useful in a combination therapy with glucocorticoids for treatment of steroid-insensitive pulmonary diseases.

  20. MiR-222 targeted PUMA to improve sensitization of UM1 cells to cisplatin.

    PubMed

    Jiang, Fangfang; Zhao, Wei; Zhou, Lijie; Liu, Zifeng; Li, Wenqing; Yu, Dongsheng

    2014-12-02

    microRNAs have been shown to play critical roles in regulating the chemosensitivity of cancer cells. As a member of the oncogenic miRNAs (oncomiRs), miR-222 has been reported to drive the oncogenesis of many types of malignancies. However, little is known concerning the specific role of miR-222 in human oral squamous cell carcinoma (OSCC). The present study explored the role and mechanism of miR-222 in increasing the expression of p53 up-regulated modulator of apoptosis (PUMA) and enhancing the sensitivity of OSCC to cisplatin (CDDP). Results showed that antisense (As)-miR-222 inhibits the expression of miR-222. In contrast, PUMA was dramaticallyup-regulated. IC50 values were significantly decreased in cells treated with As-miR-222 combined with CDDP, to a greater extent than in cells treated with CDDP alone. Furthermore, As-miR-222 enhanced apoptosis and inhibited the invasiveness of UM1 cells. Analysis of the above data suggested that, in UM1 cells, there might be a regulatory loop between miR-222 and PUMA, and that miR-222 inhibition increased the chemosensitivity to CDDP. These findings demonstrated that down-regulation of miR-222 could enhance the chemosensitivity of human OSCC cells to CDDP, and that the combination of As-miR-222 and CDDP could be an effective therapeutic strategy by boosting the expression of PUMA for controlling the growth of OSCC.

  1. Differential gene expression, GATA1 target genes, and the chemotherapy sensitivity of Down syndrome megakaryocytic leukemia.

    PubMed

    Ge, Yubin; Dombkowski, Alan A; LaFiura, Katherine M; Tatman, Dana; Yedidi, Ravikiran S; Stout, Mark L; Buck, Steven A; Massey, Gita; Becton, David L; Weinstein, Howard J; Ravindranath, Yaddanapudi; Matherly, Larry H; Taub, Jeffrey W

    2006-02-15

    Children with Down syndrome (DS) with acute megakaryocytic leukemia (AMkL) have very high survival rates compared with non-DS AMkL patients. Somatic mutations identified in the X-linked transcription factor gene, GATA1, in essentially all DS AMkL cases result in the synthesis of a shorter (40 kDa) protein (GATA1s) with altered transactivation activity and may lead to altered expression of GATA1 target genes. Using the Affymetrix U133A microarray chip, we identified 551 differentially expressed genes between DS and non-DS AMkL samples. Transcripts for the bone marrow stromal-cell antigen 2 (BST2) gene, encoding a transmembrane glycoprotein potentially involved in interactions between leukemia cells and bone marrow stromal cells, were 7.3-fold higher (validated by real-time polymerase chain reaction) in the non-DS compared with the DS group. Additional studies confirmed GATA1 protein binding and transactivation of the BST2 promoter; however, stimulation of BST2 promoter activity by GATA1s was substantially reduced compared with the full-length GATA1. CMK sublines, transfected with the BST2 cDNA and incubated with HS-5 bone marrow stromal cells, exhibited up to 1.7-fold reduced cytosine arabinoside (ara-C)-induced apoptosis, compared with mock-transfected cells. Our results demonstrate that genes that account for differences in survival between DS and non-DS AMkL cases may be identified by microarray analysis and that differential gene expression may reflect relative transactivation capacities of the GATA1s and full-length GATA1 proteins.

  2. Differential gene expression, GATA1 target genes, and the chemotherapy sensitivity of Down syndrome megakaryocytic leukemia

    PubMed Central

    Ge, Yubin; Dombkowski, Alan A.; LaFiura, Katherine M.; Tatman, Dana; Yedidi, Ravikiran S.; Stout, Mark L.; Buck, Steven A.; Massey, Gita; Becton, David L.; Weinstein, Howard J.; Ravindranath, Yaddanapudi; Matherly, Larry H.; Taub, Jeffrey W.

    2006-01-01

    Children with Down syndrome (DS) with acute megakaryocytic leukemia (AMkL) have very high survival rates compared with non-DS AMkL patients. Somatic mutations identified in the X-linked transcription factor gene, GATA1, in essentially all DS AMkL cases result in the synthesis of a shorter (40 kDa) protein (GATA1s) with altered transactivation activity and may lead to altered expression of GATA1 target genes. Using the Affymetrix U133A microarray chip, we identified 551 differentially expressed genes between DS and non-DS AMkL samples. Transcripts for the bone marrow stromal-cell antigen 2 (BST2) gene, encoding a transmembrane glycoprotein potentially involved in interactions between leukemia cells and bone marrow stromal cells, were 7.3-fold higher (validated by real-time polymerase chain reaction) in the non-DS compared with the DS group. Additional studies confirmed GATA1 protein binding and transactivation of the BST2 promoter; however, stimulation of BST2 promoter activity by GATA1s was substantially reduced compared with the full-length GATA1. CMK sublines, transfected with the BST2 cDNA and incubated with HS-5 bone marrow stromal cells, exhibited up to 1.7-fold reduced cytosine arabinoside (ara-C)-induced apoptosis, compared with mock-transfected cells. Our results demonstrate that genes that account for differences in survival between DS and non-DS AMkL cases may be identified by microarray analysis and that differential gene expression may reflect relative transactivation capacities of the GATA1s and full-length GATA1 proteins. PMID:16249385

  3. Inhibition of CRM1-dependent nuclear export sensitizes malignant cells to cytotoxic and targeted agents.

    PubMed

    Turner, Joel G; Dawson, Jana; Cubitt, Christopher L; Baz, Rachid; Sullivan, Daniel M

    2014-08-01

    the addition of alkylating agents (melphalan), anthracyclines (doxorubicin and daunomycin), BRAF inhibitors, platinum drugs (cisplatin and oxaliplatin), proteosome inhibitors (bortezomib and carfilzomib), or tyrosine-kinase inhibitors (imatinib). Also, the sequence of treatment may be important for combination therapy. We found that the most effective treatment regimen involved first priming the cancer cells with the CRM1 inhibitor followed by doxorubicin, bortezomib, carfilzomib, or melphalan. This order sensitized both de novo and acquired drug-resistant cancer cell lines.

  4. 78 FR 40144 - Governmentwide Travel Advisory Committee (GTAC); Upcoming Public Advisory Committee Meeting

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-07-03

    ... ADMINISTRATION Governmentwide Travel Advisory Committee (GTAC); Upcoming Public Advisory Committee Meeting AGENCY... Governmentwide Travel Advisory Committee (GTAC) (the Committee), is a Federal Advisory Committee established in..., Designated Federal Officer (DFO), Governmentwide Travel Advisory Committee (GTAC), Office of...

  5. Prostaglandin E2 suppresses allergic sensitization and lung inflammation by targeting the E prostanoid 2 receptor on T cells

    PubMed Central

    Zaslona, Zbigniew; Okunishi, Katsuhide; Bourdonnay, Emilie; Domingo-Gonzalez, Racquel; Moore, Bethany B.; Lukacs, Nicholas W.; Aronoff, David M.; Peters-Golden, Marc

    2013-01-01

    Background Endogenous prostanoids have been suggested to modulate sensitization during experimental allergic asthma, but the specific role of prostaglandin E2 (PGE2) or of specific E prostanoid (EP) receptors is not known. Objective Here we tested the role of EP2 signaling in allergic asthma. Methods Wild type (WT) and EP2−/− mice were subjected to ovalbumin sensitization and acute airway challenge. The PGE2 analog misoprostol was administered during sensitization in both genotypes. In vitro culture of splenocytes and of flow-sorted dendritic cells and T cells defined the mechanism by which EP2 exerted its protective effect. Adoptive transfer of WT and EP2−/− CD4 T cells was used to validate the importance of EP2 expression on T cells. Results As compared to WT mice, EP2−/− mice had exaggerated airway inflammation in this model. Splenocytes and lung lymph node cells from sensitized EP2−/− mice produced more IL-13 than did WT cells, suggesting increased sensitization. In WT but not EP2−/− mice, subcutaneous administration of a stable PGE2 analog during sensitization inhibited allergic inflammation. PGE2 decreased cytokine production and inhibited STAT6 phosphorylation by CD3/CD28-stimulated CD4pos T cells. Co-culture of flow cytometry-sorted splenic CD4pos T cells and CD11cpos dendritic cells from WT or EP2−/− mice suggested that the increased IL-13 production in EP2−/− mice was due to the lack of EP2 specifically on T cells. Adoptive transfer of CD4pos EP2−/− T cells caused greater cytokine production in the lungs of WT mice than did transfer of WT CD4pos T cells. Conclusion We conclude that the PGE2-EP2 axis is an important endogenous brake on allergic airway inflammation, primarily targets T cells, and its agonism represents a potential novel therapeutic approach to asthma. PMID:24075232

  6. 77 FR 74099 - Reestablishment of Advisory Group

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-12-12

    .... Reestablishing the Advisory Group on Prevention, Health Promotion, and Integrative and Public Health. The Advisory Group on Prevention, Health Promotion, and Integrative and Public Health (Advisory Group), as...

  7. A Nanoparticle Carrying the p53 Gene Targets Tumors Including Cancer Stem Cells, Sensitizes Glioblastoma to Chemotherapy and Improves Survival

    PubMed Central

    2015-01-01

    Temozolomide (TMZ)-resistance in glioblastoma multiforme (GBM) has been linked to upregulation of O6-methylguanine-DNA methyltransferase (MGMT). Wild-type (wt) p53 was previously shown to down-modulate MGMT. However, p53 therapy for GBM is limited by lack of efficient delivery across the blood brain barrier (BBB). We have developed a systemic nanodelivery platform (scL) for tumor-specific targeting (primary and metastatic), which is currently in multiple clinical trials. This self-assembling nanocomplex is formed by simple mixing of the components in a defined order and a specific ratio. Here, we demonstrate that scL crosses the BBB and efficiently targets GBM, as well as cancer stem cells (CSCs), which have been implicated in recurrence and treatment resistance in many human cancers. Moreover, systemic delivery of scL-p53 down-modulates MGMT and induces apoptosis in intracranial GBM xenografts. The combination of scL-p53 and TMZ increased the antitumor efficacy of TMZ with enhanced survival benefit in a mouse model of highly TMZ-resistant GBM. scL-p53 also sensitized both CSCs and bulk tumor cells to TMZ, increasing apoptosis. These results suggest that combining scL-p53 with standard TMZ treatment could be a more effective therapy for GBM. PMID:24811110

  8. On the sensitivity of the goes flare classification to properties of the electron beam in the thick-target model

    SciTech Connect

    Reep, J. W.; Bradshaw, S. J.; McAteer, R. T. J. E-mail: stephen.bradshaw@rice.edu

    2013-11-20

    The collisional thick-target model, wherein a large number of electrons are accelerated down a flaring loop, can be used to explain many observed properties of solar flares. In this study, we focus on the sensitivity of (GOES) flare classification to the properties of the thick-target model. Using a hydrodynamic model with RHESSI-derived electron beam parameters, we explore the effects of the beam energy flux (or total non-thermal energy), the cut-off energy, and the spectral index of the electron distribution on the soft X-rays observed by GOES. We conclude that (1) the GOES class is proportional to the non-thermal energy E {sup α} for α ≈ 1.7 in the low-energy passband (1-8 Å) and ≈1.6 in the high-energy passband (0.5-4 Å); (2) the GOES class is only weakly dependent on the spectral index in both passbands; (3) increases in the cut-off will increase the flux in the 0.5-4 Å passband but decrease the flux in the 1-8 Å passband, while decreases in the cut-off will cause a decrease in the 0.5-4 Å passband and a slight increase in the 1-8 Å passband.

  9. Antitumor activity of EGFR targeted pH-sensitive immunoliposomes encapsulating gemcitabine in A549 xenograft nude mice.

    PubMed

    Kim, In-Young; Kang, Young-Sook; Lee, Doo Sung; Park, Heon-Joo; Choi, Eun-Kyung; Oh, Yu-Kyoung; Son, Hye-Jung; Kim, Jin-Seok

    2009-11-16

    Immunoliposomes directed by monoclonal antibodies are promising vehicles for tumor targeted drug delivery. Development of a long-circulating formulation of pH-sensitive liposomes (PSLs) with epidermal growth factor receptor (EGFR) antibody attached was designed and tested using A549 cells and BALB/c-nu/nu mouse tumor model. PSL formulation was prepared using small unilamellar vesicles of DOPE and CHEMS (6:4 molar ratio) by REV method. The average size and zeta-potential of the formulation measured by dynamic laser-light scattering were approximately 146+/-43.9 nm (PDI=0.09+/-0.02) and -1.77+/-0.03 mV, respectively. A549 cells were xenotransplanted into BALB/c-nu/nu mice and various formulations of gemcitabine (gem), such as in its free form, PSLs or Ab-PSLs, were injected intravenously via a tail vein. The rate of tumor volume increment in Ab-PSLs with gem-treated group was remarkably slower than that of other drug-treated group. The tumor from Ab-PSLs with gem 160 mg/kg-injected group exhibited a markedly lowest account of PCNA labeled cells and had highest TUNEL-positive cells among tested. This suggests that treatment of Ab-PSLs with gem resulted in an increased apoptosis of tumor cells, leading to tumor growth inhibition. These results demonstrate that PSLs provide an efficient and targeted delivery of gemcitabine and may represent a useful new treatment approach for tumors which overexpress the EGFR.

  10. Ammonium tetrathiomolybdate treatment targets the copper transporter ATP7A and enhances sensitivity of breast cancer to cisplatin

    PubMed Central

    Wong, Ada Hang-Heng; Vazquez-Ortiz, Guelaguetza; Chen, Weiping; Xu, Xiaoling; Deng, Chu-Xia

    2016-01-01

    Cisplatin is an effective breast cancer drug but resistance often develops over prolonged chemotherapy. Therefore, we performed a candidate approach RNAi screen in combination with cisplatin treatment to identify molecular pathways conferring survival advantages. The screen identified ATP7A as a therapeutic target. ATP7A is a copper ATPase transporter responsible for intercellular movement and sequestering of cisplatin. Pharmaceutical replacement for ATP7A by ammonium tetrathiomolybdate (TM) enhanced cisplatin treatment in breast cancer cells. Allograft and xenograft models in athymic nude mice treated with cisplatin/TM exhibited retarded tumor growth, reduced accumulation of cancer stem cells and decreased cell proliferation as compared to mono-treatment with cisplatin or TM. Cisplatin/TM treatment of cisplatin-resistant tumors reduced ATP7A protein levels, attenuated cisplatin sequestering by ATP7A, increased nuclear availability of cisplatin, and subsequently enhanced DNA damage and apoptosis. Microarray analysis of gene ontology pathways that responded uniquely to cisplatin/TM double treatment depicted changes in cell cycle regulation, specifically in the G1/S transition. These findings offer the potential to combat platinum-resistant tumors and sensitize patients to conventional breast cancer treatment by identifying and targeting the resistant tumors' unique molecular adaptations. PMID:27806319

  11. Targeting of Wnt/β-Catenin by Anthelmintic Drug Pyrvinium Enhances Sensitivity of Ovarian Cancer Cells to Chemotherapy

    PubMed Central

    Zhang, Chongyuan; Zhang, Zhenge; Zhang, Shuirong; Wang, Wenrong; Hu, Ping

    2017-01-01

    Background Aberrant activation of Wnt/β-catenin has been shown to promote ovarian cancer proliferation and chemoresistance. Pyrvinium, an FDA-approved anthelmintic drug, has been identified as a potent Wnt inhibitor. Pyrvinium may sensitize ovarian cancer cells to chemotherapy. Material/Methods The effect of pyrvinium alone and its combination with paclitaxel in ovarian cancer was investigated using an in vitro culture system and in vivo xenograft models. The mechanisms of its action were also analyzed, focusing on the Wnt/β-catenin pathway. Results Pyrvinium inhibited growth and induced apoptosis of paclitaxel- and cisplatin-resistant epithelial ovarian cancer cell lines A2278/PTX and SK-OV-3. Its combination with paclitaxel was synergistic in targeting ovarian cancer cells in vitro. In 3 independent ovarian xenograft mouse models, pyrvinium alone inhibited tumor growth. More importantly, we observed significant inhibition of tumor growth throughout the treatment when using pyrvinium and paclitaxel combined. Mechanistically, pyrvinium increased the Wnt-negative regulator axin and decreased the β-catenin levels in ovarian cancer cells. In addition, pyrvinium suppressed Wnt/β-catenin-mediated transcription, as shown by the decreased mRNA levels of MYC, cyclin D, and BCL-9. In contrast, the inhibitory effects of pyrvinium were reversed by β-catenin stabilization or overexpression, demonstrating that pyrvinium acted on ovarian cancer cells via targeting the Wnt/β-catenin signaling pathway. Conclusions We demonstrated that the anthelmintic drug pyrvinium targets ovarian cancer cells through suppressing Wnt/β-catenin signaling. Our work highlights the therapeutic value of inhibiting Wnt/β-catenin in ovarian cancer. PMID:28090074

  12. Sensitivity-enhanced solid-state NMR detection of expansin’s target in plant cell walls

    PubMed Central

    Wang, Tuo; Park, Yong Bum; Caporini, Marc A.; Rosay, Melanie; Zhong, Linghao; Cosgrove, Daniel J.; Hong, Mei

    2013-01-01

    Structure determination of protein binding to noncrystalline macromolecular assemblies such as plant cell walls (CWs) poses a significant structural biology challenge. CWs are loosened during growth by expansin proteins, which weaken the noncovalent network formed by cellulose, hemicellulose, and pectins, but the CW target of expansins has remained elusive because of the minute amount of the protein required for activity and the complex nature of the CW. Using solid-state NMR spectroscopy, combined with sensitivity-enhancing dynamic nuclear polarization (DNP) and differential isotopic labeling of expansin and polysaccharides, we have now determined the functional binding target of expansin in the Arabidopsis thaliana CW. By transferring the electron polarization of a biradical dopant to the nuclei, DNP allowed selective detection of 13C spin diffusion from trace concentrations of 13C, 15N-labeled expansin in the CW to nearby polysaccharides. From the spin diffusion data of wild-type and mutant expansins, we conclude that to loosen the CW, expansin binds highly specific cellulose domains enriched in xyloglucan, whereas more abundant binding to pectins is unrelated to activity. Molecular dynamics simulations indicate short 13C-13C distances of 4–6 Å between a hydrophobic surface of the cellulose microfibril and an aromatic motif on the expansin surface, consistent with the observed NMR signals. DNP-enhanced 2D 13C correlation spectra further reveal that the expansin-bound cellulose has altered conformation and is enriched in xyloglucan, thus providing unique insight into the mechanism of CW loosening. DNP-enhanced NMR provides a powerful, generalizable approach for investigating protein binding to complex macromolecular targets. PMID:24065828

  13. Design and evaluation of novel pH-sensitive ureido-conjugated chitosan/TPP nanoparticles targeted to Helicobacter pylori.

    PubMed

    Jing, Zi-Wei; Jia, Yi-Yang; Wan, Ning; Luo, Min; Huan, Meng-Lei; Kang, Tai-Bin; Zhou, Si-Yuan; Zhang, Bang-Le

    2016-04-01

    The covalently modified ureido-conjugated chitosan/TPP multifunctional nanoparticles have been developed as targeted nanomedicine delivery system for eradication of Helicobacter pylori. H. pylori can specifically express the urea transport protein on its membrane to transport urea into cytoplasm for urease to produce ammonia, which protects the bacterium in the acid milieu of stomach. The clinical applicability of topical antimicrobial agent is needed to eradicate H. pylori in the infected fundal area. In this study, we designed and synthesized two ureido-conjugated chitosan derivatives UCCs-1 and UCCs-2 for preparation of multifunctional nanoparticles. The process was optimized in order to prepare UCCs/TPP nanoparticles for encapsulation of amoxicillin. The results showed that the amoxicillin-UCCs/TPP nanoparticles exhibited favorable pH-sensitive characteristics, which could procrastinate the release of amoxicillin at gastric acids and enable the drug to deliver and target to H. pylori at its survival region effectively. Compared with unmodified amoxicillin-chitosan/TPP nanoparticles, a more specific and effective H. pylori growth inhibition was observed for amoxicillin-UCCs/TPP nanoparticles. Drug uptake analysis tested by flow cytometry and confocal laser scanning microscopy verified that the uptake of FITC-UCCs-2/TPP nanoparticles was associated with urea transport protein on the membrane of H. pylori and reduced with the addition of urea as competitive transport substrate. These findings suggest that the multifunctional amoxicillin-loaded nanoparticles have great potential for effective therapy of H. pylori infection. They may also serve as pharmacologically effective nanocarriers for oral targeted delivery of other therapeutic drugs to treat H. pylori.

  14. Dual-targeted and pH-sensitive Doxorubicin Prodrug-Microbubble Complex with Ultrasound for Tumor Treatment

    PubMed Central

    Luo, Wanxian; Wen, Ge; Yang, Li; Tang, Jiao; Wang, Jianguo; Wang, Jihui; Zhang, Shiyu; Zhang, Li; Ma, Fei; Xiao, Liling; Wang, Ying; Li, Yingjia

    2017-01-01

    In this study, we investigated the potential of a dual-targeted pH-sensitive doxorubicin prodrug-microbubble complex (DPMC) in ultrasound (US)-assisted antitumor therapy. The doxorubicin prodrug (DP) consists of a succinylated-heparin carrier conjugated with doxorubicin (DOX) via hydrazone linkage and decorated with dual targeting ligands, folate and cRGD peptide. Combination of microbubble (MB) and DP, generated via avidin-biotin binding, promoted intracellular accumulation and improved therapeutic efficiency assisted by US cavitation and sonoporation. Aggregates of prepared DP were observed with an inhomogeneous size distribution (average diameters: 149.6±29.8 nm and 1036.2±38.8 nm, PDI: 1.0) while DPMC exhibited a uniform distribution (average diameter: 5.804±2.1 μm), facilitating its usage for drug delivery. Notably, upon US exposure, DPMC was disrupted and aggregated DP dispersed into homogeneous small-sized nanoparticles (average diameter: 128.6±42.3 nm, PDI: 0.21). DPMC could target to angiogenic endothelial cells in tumor region via αvβ3-mediated recognition and subsequently facilitate its specific binding to tumor cells mediated via recognition of folate receptor (FR) after US exposure. In vitro experiments showed higher tumor specificity and killing ability of DPMC with US than free DOX and DP for breast cancer MCF-7 cells. Furthermore, significant accumulation and specificity for tumor tissues of DPMC with US were detected using in vivo fluorescence and ultrasound molecular imaging, indicating its potential to integrate tumor imaging and therapy. In particular, through inducing apoptosis, inhibiting cell proliferation and antagonizing angiogenesis, DPMC with US produced higher tumor inhibition rates than DOX or DPMC without US in MCF-7 xenograft tumor-bearing mice while inducing no obvious body weight loss. Our strategy provides an effective platform for the delivery of large-sized or aggregated particles to tumor sites, thereby extending their

  15. Advisory Committee Handbook.

    ERIC Educational Resources Information Center

    Black Hawk Coll., Moline, IL.

    An advisory committee is generally comprised of persons outside the education profession who have specialized knowledge in a given area. The committee advises, makes recommendations, and gives service to the college and its students, instructors, and administrators. At Black Hawk College, there are four types of advisory committees: community,…

  16. 76 FR 75874 - Technology Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-12-05

    ... COMMISSION Technology Advisory Committee AGENCY: Commodity Futures Trading Commission (``CFTC''). ACTION: Notice of Meeting of Technology Advisory Committee. SUMMARY: The Technology Advisory Committee will hold..., attention: Office of the Secretary. Please use the title ``Technology Advisory Committee'' in any...

  17. 76 FR 8715 - Technology Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-02-15

    ... COMMISSION Technology Advisory Committee AGENCY: Commodity Futures Trading Commission (``CFTC''). ACTION: Notice of meeting of Technology Advisory Committee. SUMMARY: The Technology Advisory Committee will hold...: Office of the Secretary. Please use the title ``Technology Advisory Committee'' in any written...

  18. Direct detection of calmodulin tuning by ryanodine receptor channel targets using a Ca2+-sensitive acrylodan-labeled calmodulin.

    PubMed

    Fruen, Bradley R; Balog, Edward M; Schafer, Janet; Nitu, Florentin R; Thomas, David D; Cornea, Razvan L

    2005-01-11

    Calmodulin (CaM) activates the skeletal muscle ryanodine receptor (RyR1) at nanomolar Ca(2+) concentrations but inhibits it at micromolar Ca(2+) concentrations, indicating that binding of Ca(2+) to CaM may provide a molecular switch for modulating RyR1 channel activity. To directly examine the Ca(2+) sensitivity of RyR1-complexed CaM, we used an environment-sensitive acrylodan adduct of CaM. The resulting (ACR)CaM probe displayed high-affinity binding to, and Ca(2+)-dependent regulation of, RyR1 similar to that of unlabeled wild-type (WT) CaM. Upon addition of Ca(2+), (ACR)CaM exhibited a substantial (>50%) decrease in fluorescence (K(Ca) = 2.7 +/- 0.8 microM). A peptide derived from the RyR1 CaM binding domain (RyR1(3614)(-)(43)) caused an even more pronounced Ca(2+)-dependent fluorescence decrease, and a >or=10-fold leftward shift in its K(Ca) (0.2 +/- 0.1 microM). In the presence of intact RyR1 channels in SR vesicles, (ACR)CaM fluorescence spectra were distinct from those in the presence of RyR1(3614)(-)(43), although a Ca(2+)-dependent decrease in fluorescence was still observed. The K(Ca) for (ACR)CaM fluorescence in the presence of SR (0.8 +/- 0.4 microM) was greater than in the presence of RyR1(3614)(-)(43) but was consistent with functional determinations showing the conversion of (ACR)CaM from channel activator (apoCaM) to inhibitor (Ca(2+)CaM) at Ca(2+) concentrations between 0.3 and 1 microM. These results indicate that binding to RyR1 targets evokes significant changes in the CaM structure and Ca(2+) sensitivity (i.e., CaM tuning). However, changes resulting from binding of CaM to the full-length, tetrameric channels are clearly distinct from changes caused by the RyR1-derived peptide. We suggest that the Ca(2+) sensitivity of CaM when in complex with full-length channels may be tuned to respond to physiologically relevant changes in Ca(2+).

  19. The sensitive area for targeting observation associated with two types of El Niño events

    NASA Astrophysics Data System (ADS)

    Li, Xuquan; Duan, Wansuo

    2016-04-01

    The optimal forcing vector (OFV) approach is an effective method to rectify a numerical model by offsetting the tendency error of the model. Applying the OFV approach to Zebiak-Cane model, we successfully simulate 8 El Niño events after 1980 including 3 eastern Pacific (EP) ones and 5 central Pacific (CP) ones. Then we compute the conditional nonlinear optimal perturbation (CNOP) of each El Niño event which represents the fastest growing initial error of each event. It is found that the CNOP-type initial errors of different types of El Niño event have similar structures in both SSTA pattern and thermocline depth anomaly pattern. The CNOP-type errors can be classified into two types. One type has a SSTA pattern with negative anomalies in the equatorial central western Pacific, positive anomalies in the equatorial eastern Pacific, and a thermocline depth anomaly pattern with positive anomalies along the equator; while the other type presents patterns almost opposite to the former type. All these initial errors develop dramatically and make the predict results far away from the truths. This indicates that initial errors with particular patterns can cause serious uncertainty of El Nino predictions. We choose the region where SSTA errors are larger and when subtracting the initial errors in this area, the development of initial errors is significantly depressed and as a result, the predict skill of two types of El Nino events improves greatly. The region with initial errors being larger represents the sensitive area for targeting observation associated with predictions of two types of El Nino events. Increasing observations in the sensitive area is helpful for predicting which type of El Nino event will occur.

  20. Multifunctional pH-sensitive magnetic nanoparticles for simultaneous imaging, sensing and targeted intracellular anticancer drug delivery

    NASA Astrophysics Data System (ADS)

    Banerjee, Shashwat S.; Chen, Dong-Hwang

    2008-12-01

    A novel multifunctional magnetic nanocarrier was fabricated for synchronous cancer therapy and sensing. The nanocarrier, programed to display a response to environmental stimuli (pH value), was synthesized by coupling doxorubicin (DOX) to adipic dihydrazide-grafted gum arabic modified magnetic nanoparticles (ADH-GAMNP) via the hydrolytically degradable pH-sensitive hydrazone bond. The resultant nanocarrier, DOX-ADH-GAMNP, had a mean diameter of 13.8 nm and the amount of DOX coupled was about 6.52 mg g-1. Also, it exhibited pH triggered release of DOX in an acidic environment (pH 5.0) but was relatively stable at physiological pH (pH 7.4). Furthermore, both GAMNP and DOX were found to possess fluorescence properties when excited in the near-infrared region due to the two-photon absorption mechanism. The coupling of DOX to GAMNP resulted in a reversible self-quenching of fluorescence through the fluorescence resonant energy transfer (FRET) between the donor GAMNP and acceptor DOX. The release of DOX from DOX-ADH-GAMNP when exposed to acidic media indicated the recovery of fluorescence from both GAMNP and DOX. The change in the fluorescence intensity of DOX-ADH-GAMNP on the release of DOX can act as a potential sensor to sense the delivery of the drug. The analysis of zeta potential and plasmon absorbance in different pH conditions also confirmed the pH sensitivity of the product. This multifunctional nanocarrier is a significant breakthrough in developing a drug delivery vehicle that combines drug targeting as well as sensing and therapy at the same time.

  1. Multifunctional pH-sensitive magnetic nanoparticles for simultaneous imaging, sensing and targeted intracellular anticancer drug delivery.

    PubMed

    Banerjee, Shashwat S; Chen, Dong-Hwang

    2008-12-17

    A novel multifunctional magnetic nanocarrier was fabricated for synchronous cancer therapy and sensing. The nanocarrier, programed to display a response to environmental stimuli (pH value), was synthesized by coupling doxorubicin (DOX) to adipic dihydrazide-grafted gum arabic modified magnetic nanoparticles (ADH-GAMNP) via the hydrolytically degradable pH-sensitive hydrazone bond. The resultant nanocarrier, DOX-ADH-GAMNP, had a mean diameter of 13.8 nm and the amount of DOX coupled was about 6.52 mg g(-1). Also, it exhibited pH triggered release of DOX in an acidic environment (pH 5.0) but was relatively stable at physiological pH (pH 7.4). Furthermore, both GAMNP and DOX were found to possess fluorescence properties when excited in the near-infrared region due to the two-photon absorption mechanism. The coupling of DOX to GAMNP resulted in a reversible self-quenching of fluorescence through the fluorescence resonant energy transfer (FRET) between the donor GAMNP and acceptor DOX. The release of DOX from DOX-ADH-GAMNP when exposed to acidic media indicated the recovery of fluorescence from both GAMNP and DOX. The change in the fluorescence intensity of DOX-ADH-GAMNP on the release of DOX can act as a potential sensor to sense the delivery of the drug. The analysis of zeta potential and plasmon absorbance in different pH conditions also confirmed the pH sensitivity of the product. This multifunctional nanocarrier is a significant breakthrough in developing a drug delivery vehicle that combines drug targeting as well as sensing and therapy at the same time.

  2. 77 FR 13131 - Advisory Committees; Filing of Closed Meeting Reports

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-03-05

    ... Research Allergenic Products Advisory Committee Blood Products Advisory Committee Cellular, Tissue and Gene Therapies Advisory Committee Vaccines and Related Biological Products Advisory Committee Center for...

  3. Magnetic-fluorescent-targeting multifunctional aptasensorfor highly sensitive and one-step rapid detection of ochratoxin A.

    PubMed

    Wang, Chengquan; Qian, Jing; Wang, Kan; Wang, Kun; Liu, Qian; Dong, Xiaoya; Wang, Chengke; Huang, Xingyi

    2015-06-15

    A multifunctional aptasensor for highly sensitive and one-step rapid detection of ochratoxin A (OTA), has been developed using aptamer-conjugated magnetic beads (MBs) as the recognition and concentration element and a heavy CdTe quantum dots (QDs) as the label. Initially, the thiolated aptamer was conjugated on the Fe3O4@Au MBs through Au-S covalent binding. Subsequently, multiple CdTe QDs were loaded both in and on a versatile SiO2 nanocarrier to produce a large amplification factor of hybrid fluorescent nanoparticles (HFNPs) labeled complementary DNA (cDNA). The magnetic-fluorescent-targeting multifunctional aptasensor was thus fabricated by immobilizing the HFNPs onto MBs' surface through the hybrid reaction between the aptamer and cDNA. This aptasensor can be produced at large scale in a single run, and then can be conveniently used for rapid detection of OTA through a one-step incubation procedure. The presence of OTA would trigger aptamer-OTA binding, resulting in the partial release of the HFNPs into bulk solution. After a simple magnetic separation, the supernatant liquid of the above solution contained a great number of CdTe QDs produced an intense fluorescence emission. Under the optimal conditions, the fluorescence intensity of the released HFNPs was proportional to the concentration of OTA in a wide range of 15 pg mL(-1) -100 ng mL(-1) with a detection limit of 5.4 pg mL(-1) (S/N=3). This multifunctional aptasensor represents a promising path toward routine quality control of food safety, and also creates the opportunity to develop aptasensors for other targets using this strategy.

  4. Targeting Pro-Apoptotic TRAIL Receptors Sensitizes HeLa Cervical Cancer Cells to Irradiation-Induced Apoptosis

    SciTech Connect

    Maduro, John H.; Vries, Elisabeth de; Meersma, Gert-Jan; Hougardy, Brigitte; Zee, Ate G.J. van der; Jong, Steven de

    2008-10-01

    Purpose: To investigate the potential of irradiation in combination with drugs targeting the tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) death receptor (DR)4 and DR5 and their mechanism of action in a cervical cancer cell line. Methods and Materials: Recombinant human TRAIL (rhTRAIL) and the agonistic antibodies against DR4 and DR5 were added to irradiated HeLa cells. The effect was evaluated with apoptosis and cytotoxicity assays and at the protein level. Membrane receptor expression was measured with flow cytometry. Small-interfering RNA against p53, DR4, and DR5 was used to investigate their function on the combined effect. Results: rhTRAIL and the agonistic DR4 and DR5 antibodies strongly enhanced 10-Gy-induced apoptosis. This extra effect was 22%, 23%, and 29% for rhTRAIL, DR4, and DR5, respectively. Irradiation increased p53 expression and increased the membrane expression of DR5 and DR4. p53 suppression, as well as small-interfering RNA against DR5, resulted in a significant downregulation of DR5 membrane expression but did not affect apoptosis induced by irradiation and rhTRAIL. After small-interfering RNA against DR4, rhTRAIL-induced apoptosis and the additive effect of irradiation on rhTRAIL-induced apoptosis were abrogated, implicating an important role for DR4 in apoptosis induced through irradiation in combination with rhTRAIL. Conclusion: Irradiation-induced apoptosis is strongly enhanced by targeting the pro-apoptotic TRAIL receptors DR4 or DR5. Irradiation results in a p53-dependent increase in DR5 membrane expression. The sensitizing effect of rhTRAIL on irradiation in the HeLa cell line is, however especially mediated through the DR4 receptor.

  5. CD44 Receptor Targeting and Endosomal pH-Sensitive Dual Functional Hyaluronic Acid Micelles for Intracellular Paclitaxel Delivery.

    PubMed

    Liu, Yanhua; Zhou, Chengming; Wang, Wenping; Yang, Jianhong; Wang, Hao; Hong, Wei; Huang, Yu

    2016-12-05

    A novel CD44 receptor targeting and endosome pH-sensitive dual functional hyaluronic acid-deoxycholic acid-histidine (HA-DOCA-His) micellar system was designed for intracellular paclitaxel (PTX) delivery. The HA-DOCA-His micelles exhibited desirable endosome pH (5.0-6.0)-induced aggregation and deformation behavior verified by size distribution, critical micellar concentration, and zeta potential changes. The HA-DOCA-His micelles presented excellent encapsulation efficiency and loading capacity of 90.0% and 18.9% for PTX, respectively. The PTX release from HA-DOCA-His micelles was pH-dependent, with more rapid PTX release at pH 6.0 and 5.0 than those at pH 7.4 and 6.5. The cellular uptake performance of HA-DOCA-His micelles was enhanced comparing with pH-insensitive HA-DOCA micelles by qualitative and quantitative measurements. HA-DOCA-His micelles could be taken up via CD44-receptor mediated endocytosis, transported into endosomes, and triggered drug release to cytoplasm. In vitro cytotoxicity study exhibited PTX-loaded HA-DOCA-His micelles were more active in tumor cell growth inhibition in MCF-7 cells at pH 5.8 than those at pH 6.8 and pH 7.4. A superior antitumor efficacy was demonstrated with HA-DOCA-His micelles in a MCF-7 breast tumor model. These indicated that the dual functional HA-DOCA-His micelles combined targeted intracellular delivery and endosomal release strategies could be developed as a promising nanocarrier for anticancer efficacy improvement of PTX.

  6. Se-methylselenocysteine sensitizes hypoxic tumor cells to irinotecan by targeting hypoxia-inducible factor 1α

    PubMed Central

    Chintala, Sreenivasulu; Tóth, Károly; Cao, Shousong; Durrani, Farukh A.; Vaughan, Mary M.; Jensen, Randy L.; Rustum, Youcef M.

    2010-01-01

    Purpose Hypoxic tumor cells overexpressing hypoxia-inducible factor 1alpha (HIF-1α) are generally resistant to chemo/radiotherapy. We have reported that Se-methylselenocysteine (MSC) therapeutically enhances the efficacy and selectivity of irinotecan against human tumor xenografts. The aim of this study was to delineate the mechanism responsible for the observed efficacy targeting on HIF-1α and its transcriptionally regulated genes VEGF and CAIX. Methods We investigated the mechanism of HIF-1α inhibition by MSC and its critical role in the therapeutic outcome by generating HIF-1α stable knockdown (KD) human head and neck squamous cell carcinoma, FaDu by transfecting HIF-1α short hairpin RNA. Results While cytotoxic efficacy in combination with methylselenic acid (MSA) with SN-38 (active metabolites of MSC and irinotecan) could not be confirmed in vitro against normoxic tumor cells, the hypoxic tumor cells were more sensitive to the combination. Reduction in HIF-1α either by MSA or shRNA knockdown resulted in significant increase in cytotoxicity of SN38 in vitro against hypoxic, but not the normoxic tumor cells. Similarly, in vivo, either MSC in combination with irinotecan treatment of parental xenografts or HIF-1α KD tumors treated with irinotecan alone resulted in comparable therapeutic response and increase in the long-term survival of mice bearing FaDu xenografts. Conclusions Our results show that HIF-1α is a critical target for MSC and its inhibition was associated with enhanced antitumor activity of irinotecan. Inhibition of HIF-1α appeared to be mediated through stabilization of PHD2, 3 and downregulation of ROS by MSC. Thus, our findings support the development of MSC as a HIF-1α inhibitor in combination chemotherapy. PMID:20066420

  7. WP1066 Sensitizes Oral Squamous Cell Carcinoma Cells to Cisplatin by Targeting STAT3/miR-21 axis

    PubMed Central

    Zhou, Xuan; Ren, Yu; Liu, Aiqin; Jin, Rui; Jiang, Qingping; Huang, Yuanyuan; Kong, Lingping; Wang, Xudong; Zhang, Lun

    2014-01-01

    Accumulating evidence reveals that activation of STAT3 and miR-21 contributes to chemoresistance in multiple tumors. We examined the expression of STAT3 and miR-21 in 43 oral squamous cell carcinoma (OSCC) tumors and classified them into cisplatin sensitive or resistant group. Tca8113 and Tca8113/DDP cells were treated with cisplatin (DDP), WP1066 (STAT3 inhibitor) or in combination. MTT, colony formation, wound healing, 3-D culture, and transwell chamber assays were used to evaluate the malignant phenotype of OSCC cells. We evaluated the effect of WP1066 on the expression of STAT3 and miR-21. A Tca8113/DDP OSCC xenograft tumor model was established to evaluate the therapeutic effect of WP1066 in combination with DDP. The expression of STAT3/miR-21 was significantly increased in DDP-resistant OSCC samples and Tca8113/DDP cells compared to its parental cell. Treatment of DDP combined with WP1066 efficiently inhibited Tca8113 and Tca8113/DDP cell proliferation, migration and invasion. STAT3 mediated OSCC cell survival and DDP resistance through upregulating the expression of miR-21 and downregulating miR-21 downstream targets, including PTEN, TIMP3 and PDCD4. WP1066 plus DDP treatment could inhibit Tca8113 and Tca8113/DDP cell growth by inhibiting STAT3 phosphorylation and miR-21 expression. These results indicated that STAT3/miR-21 axis could be a candidate therapeutic target for OSCC chemoresistance. PMID:25514838

  8. Zyflamend Suppresses Growth and Sensitizes Human Pancreatic Tumors to Gemcitabine in an Orthotopic Mouse Model Through Modulation of Multiple Targets

    PubMed Central

    Kunnumakkara, Ajaikumar B.; Sung, Bokyung; Ravindran, Jayaraj; Diagaradjane, Parmeswaran; Deorukhkar, Amit; Dey, Sanjit; Koca, Cemile; Tong, Zhimin; Gelovani, Juri G.; Guha, Sushovan; Krishnan, Sunil; Aggarwal, Bharat B.

    2011-01-01

    Agents that can potentiate the efficacy of standard chemotherapy against pancreatic cancer are of great interest. Because of their low cost and safety, patients commonly use a variety of dietary supplements, although evidence of their efficacy is often lacking. One such commonly used food supplement, Zyflamend, is a polyherbal preparation with potent anti-inflammatory activities, and preclinical efficacy against prostate and oral cancer. Whether Zyflamend has any efficacy against human pancreatic cancer alone or in combination with gemcitibine, a commonly used agent, was examined in cell cultures and in an orthotopic mouse model. In vitro, Zyflamend inhibited the proliferation of pancreatic cancer cell lines regardless of p53 status and also enhanced gemcitabine-induced apoptosis. This finding correlated with inhibition of NF-κB activation by Zyflamend and suppression of cyclin D1, c-myc, COX-2, Bcl-2, IAP, survivin, VEGF, ICAM-1, and CXCR4. In nude mice, oral administration of Zyflamend alone significantly inhibited the growth of orthotopically transplanted human pancreatic tumors, and when combined with gemcitabine, further enhanced the antitumor effects. Immunohistochemical and Western blot analyses of tumor tissue showed that the suppression of pancreatic cancer growth correlated with inhibition of proliferation index marker (Ki-67), COX-2, MMP-9, NF-κB, and VEGF. Overall, these results suggest that the concentrated multiherb product Zyflamend alone can inhibit the growth of human pancreatic tumors and, in addition, can sensitize pancreatic cancers to gemcitabine through the suppression of multiple targets linked to tumorigenesis. PMID:21935918

  9. Targeting miR-21 enhances the sensitivity of human colon cancer HT-29 cells to chemoradiotherapy in vitro.

    PubMed

    Deng, Jun; Lei, Wan; Fu, Jian-Chun; Zhang, Ling; Li, Jun-He; Xiong, Jian-Ping

    2014-01-17

    5-Fluorouracil (5-FU) is a classic chemotherapeutic drug that has been widely used for colorectal cancer treatment, but colorectal cancer cells are often resistant to primary or acquired 5-FU therapy. Several studies have shown that miR-21 is significantly elevated in colorectal cancer. This suggests that this miRNA might play a role in this resistance. In this study, we investigated this possibility and the possible mechanism underlying this role. We showed that forced expression of miR-21 significantly inhibited apoptosis, enhanced cell proliferation, invasion, and colony formation ability, promoted G1/S cell cycle transition and increased the resistance of tumor cells to 5-FU and X radiation in HT-29 colon cancer cells. Furthermore, knockdown of miR-21 reversed these effects on HT-29 cells and increased the sensitivity of HT-29/5-FU to 5-FU chemotherapy. Finally, we showed that miR-21 targeted the human mutS homolog2 (hMSH2), and indirectly regulated the expression of thymidine phosphorylase (TP) and dihydropyrimidine dehydrogenase (DPD). These results demonstrate that miR-21 may play an important role in the 5-FU resistance of colon cancer cells.

  10. Proton-sensitive cation channels and ion exchangers in ischemic brain injury: new therapeutic targets for stroke?

    PubMed Central

    Leng, Tiandong; Shi, Yejie; Xiong, Zhi-Gang; Sun, Dandan

    2014-01-01

    Ischemic brain injury results from complicated cellular mechanisms. The present therapy for acute ischemic stroke is limited to thrombolysis with the recombinant tissue plasminogen activator (rtPA) and mechanical recanalization. Therefore, a better understanding of ischemic brain injury is needed for the development of more effective therapies. Disruption of ionic homeostasis plays an important role in cell death following cerebral ischemia. Glutamate receptor-mediated ionic imbalance and neurotoxicity have been well established in cerebral ischemia after stroke. However, non-NMDA receptor-dependent mechanisms, involving acid-sensing ion channel 1a (ASIC1a), transient receptor potential melastatin 7 (TRPM7), and Na+/H+ exchanger isoform 1 (NHE1), have recently emerged as important players in the dysregulation of ionic homeostasis in the CNS under ischemic conditions. These H+-sensitive channels and/or exchangers are expressed in the majority of cell types of the neurovascular unit. Sustained activation of these proteins causes excessive influx of cations, such as Ca2+, Na+, and Zn2+, and leads to ischemic reperfusion brain injury. In this review, we summarize recent pre-clinical experimental research findings on how these channels/exchangers are regulated in both in vitro and in vivo models of cerebral ischemia. The blockade or transgenic knockdown of these proteins was shown to be neuroprotective in these ischemia models. Taken together, these non-NMDA receptor-dependent mechanisms may serve as novel therapeutic targets for stroke intervention. PMID:24467911

  11. MicroRNA-320a sensitizes tamoxifen-resistant breast cancer cells to tamoxifen by targeting ARPP-19 and ERRγ.

    PubMed

    Lü, Mingrong; Ding, Keshuo; Zhang, Guofeng; Yin, Mianmian; Yao, Guidong; Tian, Hui; Lian, Jie; Liu, Lin; Liang, Meng; Zhu, Tao; Sun, Fei

    2015-03-04

    Tamoxifen represents a major adjuvant therapy to those patients with estrogen receptor-alpha positive breast cancer. However, tamoxifen resistance occurs quite often, either de novo or acquired during treatment. To investigate the role of miR-320a in the development of resistance to tamoxifen, we established tamoxifen-resistant (TamR) models by continually exposing MCF-7 or T47D breast cancer cells to tamoxifen, and identified microRNA(miRNA)-320a as a down-regulated miRNA in tamoxifen resistant cells. Re-expression of miR-320a was sufficient to sensitize TamR cells to tamoxifen by targeting cAMP-regulated phosphoprotein (ARPP-19) and estrogen-related receptor gamma (ERRγ) as well as their downstream effectors, c-Myc and Cyclin D1. Furthermore, progesterone (P4) promoted the expression of miR-320a by repressing c-Myc expression, while estrogen (E2) exerted the opposite effect. These results suggest the potential therapeutic approach for tamoxifen-resistant breast cancer by restorating miR-320a expression or depleting ARPP-19/ERRγ expression.

  12. Tumor-targeting, pH-sensitive nanoparticles for docetaxel delivery to drug-resistant cancer cells

    PubMed Central

    Tran, Tuan Hiep; Ramasamy, Thiruganesh; Choi, Ju Yeon; Nguyen, Hanh Thuy; Pham, Thanh Tung; Jeong, Jee-Heon; Ku, Sae Kwang; Choi, Han-Gon; Yong, Chul Soon; Kim, Jong Oh

    2015-01-01

    The attachment of polyethylene glycol (PEG) increases the circulation time of drug-containing nanoparticles; however, this also negatively affects cellular uptake. To overcome this problem, unique lipid polymer hybrid (LPH) nanoparticles were developed with a pH-responsive PEG layer that detached prior to cell uptake. Docetaxel (DTX) was incorporated into the lipid core of the nanoparticles, which was then shielded with the pH-responsive block co-polymer polyethylene glycol-b-polyaspartic acid (PEG-b-PAsp) using a modified emulsion method. The optimized LPH nanoparticles were ~200 nm and had a narrow size distribution. Drug release from DTX-loaded LPH (DTX-LPH) nanoparticles was pH-sensitive, which is beneficial for tumor targeting. More importantly, DTX-LPH nanoparticles were able to effectively induce apoptosis in cancer cells. The negative surface charge and PEG shell of vehicle remarkably enhanced the blood circulation and physiological activity of DTX-LPH nanoparticles compared with that of free DTX. The nanoparticles were also found to reduce the size of tumors in tumor-bearing xenograft mice. The in vivo anticancer effect of DTX-LPH nanoparticles was further confirmed by the elevated levels of caspase-3 and poly ADP ribose polymerase found in the tumors after treatment. Thus, the results suggest that this novel LPH system could be an effective new treatment for cancer. PMID:26346426

  13. 76 FR 22395 - Federal Advisory Committee Act; Open Internet Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-04-21

    ... COMMISSION Federal Advisory Committee Act; Open Internet Advisory Committee AGENCY: Federal Communications... ``Open Internet Advisory Committee'' (hereinafter ``the Committee''), is being established. FOR FURTHER... Internet rules (available at...

  14. Improving sensitivity and specificity of capturing and detecting targeted cancer cells with anti-biofouling polymer coated magnetic iron oxide nanoparticles.

    PubMed

    Lin, Run; Li, Yuancheng; MacDonald, Tobey; Wu, Hui; Provenzale, James; Peng, Xingui; Huang, Jing; Wang, Liya; Wang, Andrew Y; Yang, Jianyong; Mao, Hui

    2017-02-01

    Detecting circulating tumor cells (CTCs) with high sensitivity and specificity is critical to management of metastatic cancers. Although immuno-magnetic technology for in vitro detection of CTCs has shown promising potential for clinical applications, the biofouling effect, i.e., non-specific adhesion of biomolecules and non-cancerous cells in complex biological samples to the surface of a device/probe, can reduce the sensitivity and specificity of cell detection. Reported herein is the application of anti-biofouling polyethylene glycol-block-allyl glycidyl ether copolymer (PEG-b-AGE) coated iron oxide nanoparticles (IONPs) to improve the separation of targeted tumor cells from aqueous phase in an external magnetic field. PEG-b-AGE coated IONPs conjugated with transferrin (Tf) exhibited significant anti-biofouling properties against non-specific protein adsorption and off-target cell uptake, thus substantially enhancing the ability to target and separate transferrin receptor (TfR) over-expressed D556 medulloblastoma cells. Tf conjugated PEG-b-AGE coated IONPs exhibited a high capture rate of targeted tumor cells (D556 medulloblastoma cell) in cell media (58.7±6.4%) when separating 100 targeted tumor cells from 1×10(5) non-targeted cells and 41 targeted tumor cells from 100 D556 medulloblastoma cells spiked into 1mL blood. It is demonstrated that developed nanoparticle has higher efficiency in capturing targeted cells than widely used micron-sized particles (i.e., Dynabeads(®)).

  15. 78 FR 67362 - Federal Advisory Committee Act; Technological Advisory Council

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-11-12

    .... FOR FURTHER INFORMATION CONTACT: Walter Johnston, Chief, Electromagnetic Compatibility Division, 202-418-0807; Walter.Johnston@FCC.gov . SUPPLEMENTARY INFORMATION: The FCC Technological Advisory Council... to: Walter Johnston, the FCC's Designated Federal Officer for Technological Advisory Council by...

  16. 77 FR 12839 - Federal Advisory Committee Act; Technological Advisory Council

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-03-02

    ... 20554. FOR FURTHER INFORMATION CONTACT: Walter Johnston, Chief, Electromagnetic Compatibility Division, 202-418-0807; Walter.Johnston@FCC.gov . SUPPLEMENTARY INFORMATION: Technical Advisory Council members... Johnston, the FCC's Designated Federal Officer for Technological Advisory Council by email:...

  17. 78 FR 33092 - Federal Advisory Committee Act; Technological Advisory Council

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-06-03

    ... From the Federal Register Online via the Government Publishing Office FEDERAL COMMUNICATIONS COMMISSION Federal Advisory Committee Act; Technological Advisory Council AGENCY: Federal Communications..., this notice advises interested persons that the Federal Communications Commission's (FCC)...

  18. Rockford's Remarkable Advisory Council

    ERIC Educational Resources Information Center

    Mitchell, Brad

    1977-01-01

    Describes the cooperative relationship between the Rockford, Illinois, advisory council and the Comprehensive Employment and Training Act (CETA) staff for Winnebago and Boone Counties and credits this cooperation and community input with CETA's success. (MF)

  19. Environmental Laboratory Advisory Board

    EPA Pesticide Factsheets

    The Environmental Laboratory Advisory Board (ELAB) was established to provide consensus advice, information and recommendations on issues related to EPA measurement programs, and operation of the national accreditation program

  20. FIFRA Scientific Advisory Panel

    EPA Pesticide Factsheets

    Experts on the Federal Insecticide, Fungicide, and Rodenticide Act Scientific Advisory Panel provide independent scientific advice to the EPA on a wide range of health and safety issues related to pesticides.

  1. Automated Pilot Advisory System

    NASA Technical Reports Server (NTRS)

    Parks, J. L., Jr.; Haidt, J. G.

    1981-01-01

    An Automated Pilot Advisory System (APAS) was developed and operationally tested to demonstrate the concept that low cost automated systems can provide air traffic and aviation weather advisory information at high density uncontrolled airports. The system was designed to enhance the see and be seen rule of flight, and pilots who used the system preferred it over the self announcement system presently used at uncontrolled airports.

  2. Determination of the primary target of a quinolone drug and the effect of quinolone resistance-conferring mutations by measuring quinolone sensitivity based on its mode of action.

    PubMed

    Pfeiffer, Emily S; Hiasa, Hiroshi

    2007-09-01

    We used an assay to measure quinolone sensitivity as a shift in the position of the cleavage-religation equilibrium. This assay was found to be useful in identifying the primary target of a quinolone drug and assessing the effect of quinolone resistance-conferring mutations.

  3. 78 FR 69991 - Advisory Committee; Veterinary Medicine Advisory Committee; Termination

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-11-22

    ... HUMAN SERVICES Food and Drug Administration 21 CFR Part 14 Advisory Committee; Veterinary Medicine... Food and Drug Administration (FDA) is announcing the termination of the Veterinary Medicine Advisory Committee. This document removes the Veterinary Advisory Committee from the Agency's list of...

  4. MiR-490-3p sensitizes ovarian cancer cells to cisplatin by directly targeting ABCC2

    PubMed Central

    Tian, Jing; Xu, Yan-Ying; Li, Lian; Hao, Quan

    2017-01-01

    Cisplatin (CDDP) resistance becomes a large obstacle of the beneficial therapy for patients with ovarian cancer. MicroRNAs (miRNAs) act as post-transcriptional regulators of multiple genes’ expression and have been reported to be involved in multi-drug resistance. The purpose of this study was to determine the roles and molecular mechanism of miR-490-3p in the CDDP resistance in ovarian cancer. We found that miR-490-3p was downregulated in CDDP-resistant OVCAR3/CDDP and SKOV3/CDDP cells, which was due to the hypermethylation of miR-490-3p promoter. Functional studies demonstrated that miR-490-3p increased the cell response to CDDP in OVCAR3, SKOV3 and CDDP-resistant cells, while miR-490-3p inhibition resulted in opposite effects. Luciferase assay, real-time PCR and Western blot as well as immunohistochemistry validated that ABCC2 was a direct target of miR-490-3p and miR-490-3p downregulated ABCC2 expression via binding to its 3’UTR. Importantly, silencing of ABCC2 alleviated CDDP resistance induced by miR-490-3p inhibition, while ABCC2 overexpression restored CDDP resistance inhibited by miR-490-3p. In vivo study showed that miR-490-3p enhanced the cytotoxicity of CDDP. Finally, we found that miR-490-3p was downregulated in CDDP-resistant ovarian cancer tissues, while ABCC2 was upregulated. In conclusion, our data indicate that miR-490-3p enhances CDDP sensitivity of ovarian cancer cells through downregulating ABCC2 expression, and suggest that delivery of miR-490-3p might be a potential therapeutic strategy for patients with CDP-resistant ovarian cancer. PMID:28386339

  5. Targeting neddylation induces DNA damage and checkpoint activation and sensitizes chronic lymphocytic leukemia B cells to alkylating agents.

    PubMed

    Paiva, C; Godbersen, J C; Berger, A; Brown, J R; Danilov, A V

    2015-07-09

    Microenvironment-mediated upregulation of the B-cell receptor (BCR) and nuclear factor-κB (NF-κB) signaling in CLL cells resident in the lymph node and bone marrow promotes apoptosis evasion and clonal expansion. We recently reported that MLN4924 (pevonedistat), an investigational agent that inhibits the NEDD8-activating enzyme (NAE), abrogates stromal-mediated NF-κB pathway activity and CLL cell survival. However, the NAE pathway also assists degradation of multiple other substrates. MLN4924 has been shown to induce DNA damage and cell cycle arrest, but the importance of this mechanism in primary neoplastic B cells has not been studied. Here we mimicked the lymph node microenvironment using CD40 ligand (CD40L)-expressing stroma and interleukin-21 (IL-21) to find that inducing proliferation of the primary CLL cells conferred enhanced sensitivity to NAE inhibition. Treatment of the CD40-stimulated CLL cells with MLN4924 resulted in deregulation of Cdt1, a DNA replication licensing factor, and cell cycle inhibitors p21 and p27. This led to DNA damage, checkpoint activation and G2 arrest. Alkylating agents bendamustine and chlorambucil enhanced MLN4924-mediated DNA damage and apoptosis. These events were more prominent in cells stimulated with IL-21 compared with CD40L alone, indicating that, following NAE inhibition, the culture conditions were able to direct CLL cell fate from an NF-κB inhibition to a Cdt1 induction program. Our data provide insight into the biological consequences of targeting NAE in CLL and serves as further rationale for studying the clinical activity of MLN4924 in CLL, particularly in combination with alkylating agents.

  6. Targeting helicase-dependent amplification products with an electrochemical genosensor for reliable and sensitive screening of genetically modified organisms.

    PubMed

    Moura-Melo, Suely; Miranda-Castro, Rebeca; de-Los-Santos-Álvarez, Noemí; Miranda-Ordieres, Arturo J; Dos Santos Junior, J Ribeiro; da Silva Fonseca, Rosana A; Lobo-Castañón, Maria Jesús

    2015-08-18

    Cultivation of genetically modified organisms (GMOs) and their use in food and feed is constantly expanding; thus, the question of informing consumers about their presence in food has proven of significant interest. The development of sensitive, rapid, robust, and reliable methods for the detection of GMOs is crucial for proper food labeling. In response, we have experimentally characterized the helicase-dependent isothermal amplification (HDA) and sequence-specific detection of a transgene from the Cauliflower Mosaic Virus 35S Promoter (CaMV35S), inserted into most transgenic plants. HDA is one of the simplest approaches for DNA amplification, emulating the bacterial replication machinery, and resembling PCR but under isothermal conditions. However, it usually suffers from a lack of selectivity, which is due to the accumulation of spurious amplification products. To improve the selectivity of HDA, which makes the detection of amplification products more reliable, we have developed an electrochemical platform targeting the central sequence of HDA copies of the transgene. A binary monolayer architecture is built onto a thin gold film where, upon the formation of perfect nucleic acid duplexes with the amplification products, these are enzyme-labeled and electrochemically transduced. The resulting combined system increases genosensor detectability up to 10(6)-fold, allowing Yes/No detection of GMOs with a limit of detection of ∼30 copies of the CaMV35S genomic DNA. A set of general utility rules in the design of genosensors for detection of HDA amplicons, which may assist in the development of point-of-care tests, is also included. The method provides a versatile tool for detecting nucleic acids with extremely low abundance not only for food safety control but also in the diagnostics and environmental control areas.

  7. Cytomembrane ATP-sensitive K+ channels in neurovascular unit targets of ischemic stroke in the recovery period

    PubMed Central

    Zhang, Yang; Pan, Sipei; Zheng, Xiaolu; Wan, Qi

    2016-01-01

    The present study was to analyze the mechanism of cytomembrane ATP-sensitive K+ channels (KATP) in the neurovascular unit treatment of ischemic stroke in the recovery period. A total of 24 healthy adult male Wistar rats of 5–8 weeks age, weighing 160–200 g were randomly divided into the control (sham-operation group), model, KATP blocker and KATP opener groups (n=6 rats per group). Nylon cerebral artery occlusion was conducted using nylon monofilament coated with Poly-L-lysine, which was used to produce a cerebral infarction model. After feeding normally for 3 days, 5-hydroxydecanoate (40 mg/Kg), and diazoxide (40 mg/Kg) were injected to the abdominal cavity in the blocker, and opener groups, respectively. The control received an equivalent normal saline that was injected into the sham-operation and model groups. The animals were mutilated and samples were collected after 3 days. RT-PCR was used to detect the expression levels of the three subunits of KATP, i.e., kir6.1, and sulfonylurea receptor (SUR) 1 and SUR2 mRNA, as well as to calculate infarct size in tetrazolium chloride staining. The expression level of mRNA in the opener group were significantly higher, followed by the model and blocker groups, with the control group being the lowest (P<0.05). Infarct size in the opener group was markedly smaller than the model and blocker groups, and infarct size in the blocker group was significantly larger (P<0.05). Thus, the target treatment on KATP may improve the prognosis of ischemic stroke during the recovery period. PMID:27446320

  8. Strand Invasion Based Amplification (SIBA®): a novel isothermal DNA amplification technology demonstrating high specificity and sensitivity for a single molecule of target analyte.

    PubMed

    Hoser, Mark J; Mansukoski, Hannu K; Morrical, Scott W; Eboigbodin, Kevin E

    2014-01-01

    Isothermal nucleic acid amplification technologies offer significant advantages over polymerase chain reaction (PCR) in that they do not require thermal cycling or sophisticated laboratory equipment. However, non-target-dependent amplification has limited the sensitivity of isothermal technologies and complex probes are usually required to distinguish between non-specific and target-dependent amplification. Here, we report a novel isothermal nucleic acid amplification technology, Strand Invasion Based Amplification (SIBA). SIBA technology is resistant to non-specific amplification, is able to detect a single molecule of target analyte, and does not require target-specific probes. The technology relies on the recombinase-dependent insertion of an invasion oligonucleotide (IO) into the double-stranded target nucleic acid. The duplex regions peripheral to the IO insertion site dissociate, thereby enabling target-specific primers to bind. A polymerase then extends the primers onto the target nucleic acid leading to exponential amplification of the target. The primers are not substrates for the recombinase and are, therefore unable to extend the target template in the absence of the IO. The inclusion of 2'-O-methyl RNA to the IO ensures that it is not extendible and that it does not take part in the extension of the target template. These characteristics ensure that the technology is resistant to non-specific amplification since primer dimers or mis-priming are unable to exponentially amplify. Consequently, SIBA is highly specific and able to distinguish closely-related species with single molecule sensitivity in the absence of complex probes or sophisticated laboratory equipment. Here, we describe this technology in detail and demonstrate its use for the detection of Salmonella.

  9. Exonuclease III-Assisted Target Recycling Amplification Coupled with Liposome-Assisted Amplification: One-Step and Dual-Amplification Strategy for Highly Sensitive Fluorescence Detection of DNA.

    PubMed

    Zhou, Fulin; Li, Baoxin

    2015-07-21

    Detection of ultralow concentration of specific DNA sequence is a central challenge in the early diagnosis of gene-related disease and biodefense application. Herein, we report a dual-amplification strategy for highly sensitive fluorescence detection of DNA. In this proposed strategy, a dumbbell-shaped DNA probe is designed to integrate target binding, magnetic separation, and signal response. In the presence of specific DNA target, the multifunctional dumbbell probe can initiate exonuclease III (Exo III)-aided target recycling amplification, and, in the meantime, generate a large number of fluorescein (FAM)-encapsulated liposomes. The developed method offers very high sensitivity due to primary amplification via numerous FAM from a liposome and secondary amplification via target recycling amplification. The detection limit of the proposed method can reach 4 aM, which is much lower than that of the Exo III-aided target recycling technique applied for DNA quantification without FAM-encapsulated liposomes amplification. Moreover, the dual-signal amplification process can be completed one-step in this system. Therefore, this method provides a simple, isothermal, and low-cost approach for sensitive detection of DNA and holds a great potential for early diagnosis in gene-related diseases.

  10. 76 FR 18757 - Monthly Public Meetings of the Local Government Advisory Committee's Small Community Advisory...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-04-05

    ... From the Federal Register Online via the Government Publishing Office ENVIRONMENTAL PROTECTION AGENCY Monthly Public Meetings of the Local Government Advisory Committee's Small Community Advisory... Advisory Committee Act, the U.S. Environmental Protection Agency's Local Government Advisory...

  11. 76 FR 29752 - The President's Management Advisory Board (PMAB); Notification of Upcoming Public Advisory Meeting

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-05-23

    ... Technology Management in the Federal Government, and Senior Executive Service (SES) Development and... ADMINISTRATION The President's Management Advisory Board (PMAB); Notification of Upcoming Public Advisory Meeting.... SUMMARY: The President's Management Advisory Board, a Federal Advisory Committee established in...

  12. 75 FR 22790 - Science Advisory Board Staff Office; Request for Nominations of Candidates for EPA's Advisory...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-04-30

    ... AGENCY Science Advisory Board Staff Office; Request for Nominations of Candidates for EPA's Advisory Council on Clean Air Compliance Analysis (Council) EPA's Clean Air Scientific Advisory Committee (CASAC) and EPA's Science Advisory Board (SAB) AGENCY: Environmental Protection Agency (EPA). ACTION:...

  13. Doses to radiation sensitive organs and structures located outside the radiotherapeutic target volume for four treatment situations

    SciTech Connect

    Foo, M.L.; McCullough, E.C.; Foote, R.L.; Pisansky, T.M.; Shaw, E.G. )

    1993-09-20

    This study documents dosage to radiation sensitive organs/structures located outside the radiotherapeutic target volume for four treatment situations: (a) head and neck, (b) brain (pituitary and temporal lobe), (c) breast and (d) pelvis. Clinically relevant treatment fields were simulated on a tissue-equivalent anthropomorphic phantom and subsequently irradiated with Cobalt-60 gamma rays, 6- and 18-MV x-ray beams. Thermoluminescent dosimeters and diodes were used to measure absorbed dose. The head and neck treatment resulted in significant doses of radiation to the lens and thyroid gland. The total treatment lens dose (300-400 cGy) could be cataractogenic while measured thyroid doses (1000-8000 cGy) have the potential of causing chemical hypothyroidism, thyroid neoplasms, Graves' disease and hyperparathyroidism. Total treatment retinal (400-700 cGy) and pituitary (460-1000 cGy) doses are below that considered capable of producing chronic disease. The pituitary treatment studied consisted of various size parallel opposed lateral and vertex fields (4 x 4 through 8 x 8 cm). The lens dose (40-200 cGy) with all field sizes is below those of clinical concern. Parotid doses (130-1200 cGy) and thyroid doses (350-600 cGy) are in a range where temporary xerostomia (parotid) and thyroid neoplasia development are a reasonable possibility. The retinal dose (4000 cGy) from the largest field size (8 x 8 cm[sup 2]) is in the range where retinopathy has been reported. The left temporal lobe treatment also used parallel opposed lateral and vertex fields (7 x 7 and 10 x 10 cm). Doses to the pituitary gland (5200-6200 cGy), both parotids (200-6900 cGy), left lens (200-300 cGy), and left retina (1700-4500 cGy) are capable of causing significant future clinical problems. Right-sided structures received insignificant doses. Secondary malignancies could result from the measured total treatment thyroid doses (670-980 cGy). 82 refs., 7 figs., 5 tabs.

  14. Target recycling amplification for label-free and sensitive colorimetric detection of adenosine triphosphate based on un-modified aptamers and DNAzymes.

    PubMed

    Gong, Xue; Li, Jinfu; Zhou, Wenjiao; Xiang, Yun; Yuan, Ruo; Chai, Yaqin

    2014-05-30

    Based on target recycling amplification, the development of a new label-free, simple and sensitive colorimetric detection method for ATP by using un-modified aptamers and DNAzymes is described. The association of the model target molecules (ATP) with the corresponding aptamers of the dsDNA probes leads to the release of the G-quadruplex sequences. The ATP-bound aptamers can be further degraded by Exonuclease III to release ATP, which can again bind the aptamers of the dsDNA probes to initiate the target recycling amplification process. Due to this target recycling amplification, the amount of the released G-quadruplex sequences is significantly enhanced. Subsequently, these G-quadruplex sequences bind hemin to form numerous peroxidase mimicking DNAzymes, which cause substantially intensified color change of the probe solution for highly sensitive colorimetric detection of ATP down to the sub-nanomolar (0.33nM) level. Our method is highly selective toward ATP against other control molecules and can be performed in one single homogeneous solution, which makes our sensing approach hold great potential for sensitive colorimetric detection of other small molecules and proteins.

  15. Well-Defined Redox-Sensitive Polyethene Glycol-Paclitaxel Prodrug Conjugate for Tumor-Specific Delivery of Paclitaxel Using Octreotide for Tumor Targeting.

    PubMed

    Yin, Tingjie; Wu, Qu; Wang, Lei; Yin, Lifang; Zhou, Jianping; Huo, Meirong

    2015-08-03

    A redox-sensitive prodrug, octreotide(Phe)-polyethene glycol-disulfide bond-paclitaxel [OCT(Phe)-PEG-ss-PTX], was successfully developed for targeted intracellular delivery of PTX. The formulation emphasizes long-circulation-time polymer-drug conjugates, combined targeting based on EPR and OCT-receptor mediated endocytosis, sharp redox response, and programmed drug release. The nontargeted redox-sensitive prodrug, mPEG-ss-PTX, and the targeted insensitive prodrug, OCT(Phe)-PEG-PTX, were also synthesized as controls. These polymer-PTX conjugates, structurally confirmed by 1H NMR, exhibited approximately 23,000-fold increase in water solubility over parent PTX and possessed drug contents ranging from 11% to 14%. The redox-sensitivity of the objective OCT(Phe)-PEG-ss-PTX prodrug was verified by in vitro PTX release profile in simulated reducing conditions, and the SSTRs-mediated endocytosis was demonstrated by flow cytometry and confocal laser scanning microscopy analyses. Consequently, compared with mPEG-PTX and OCT(Phe)-PEG-PTX, the OCT(Phe)-PEG-ss-PTX exhibited much stronger cyotoxicity and apoptosis-inducing ability against NCI-H446 tumor cells (SSTRs overexpression), whereas a comparable cytotoxicity of these prodrugs was obtained against WI-38 normal cells (no SSTRs expression). Finally, the in vivo studies on NCI-H466 tumor-bearing nude mice demonstrated that the OCT(Phe)-PEG-ss-PTX possessed superior tumor-targeting ability and antitumor activity over mPEG-PTX, OCT(Phe)-PEG-PTX and Taxol, as well as minimal collateral damage. This targeted redox-sensitive polymer-PTX prodrug system is promising in tumor therapy.

  16. Differential sensitivity of 16S rRNA targeted oligonucleotide probes used for fluorescence in situ hybridization is a result of ribosomal higher order structure.

    PubMed

    Frischer, M E; Floriani, P J; Nierzwicki-Bauer, S A

    1996-10-01

    The use of 16S rRNA targeted gene probes for the direct analysis of microbial communities has revolutionized the field of microbial ecology, yet a comprehensive approach for the design of such probes does not exist. The development of 16S rRNA targeted oligonucleotide probes for use with fluorescence in situ hybridization (FISH) procedures has been especially difficult as a result of the complex nature of the rRNA target molecule. In this study a systematic comparison of 16S rRNA targeted oligonucleotide gene probes was conducted to determine if target location influences the hybridization efficiency of oligonucleotide probes when used with in situ hybridization protocols for the detection of whole microbial cells. Five unique universal 12-mer oligonucleotide sequences, located at different regions of the 16S rRNA molecule, were identified by a computer-aided sequence analysis of over 1000 partial and complete 16S rRNA sequences. The complements of these oligomeric sequences were chemically synthesized for use as probes and end labeled with either [gamma-32P]ATP or the fluorescent molecule tetramethylrhodamine-5/-6. Hybridization sensitivity for each of the probes was determined by hybridization to heat-denatured RNA immobilized on blots or to formaldehyde fixed whole cells. All of the probes hybridized with equal efficiency to denatured RNA. However, the probes exhibited a wide range of sensitivity (from none to very strong) when hybridized with whole cells using a previously developed FISH procedure. Differential hybridization efficiencies against whole cells could not be attributed to cell wall type, since the relative probe efficiency was preserved when either Gram-negative or -positive cells were used. These studies represent one of the first attempts to systematically define criteria for 16S rRNA targeted probe design for use against whole cells and establish target site location as a critical parameter in probe design.

  17. Enzyme-free and label-free fluorescence aptasensing strategy for highly sensitive detection of protein based on target-triggered hybridization chain reaction amplification.

    PubMed

    Wang, Xiuzhong; Jiang, Aiwen; Hou, Ting; Li, Haiyin; Li, Feng

    2015-08-15

    Proteins are of great importance in medical and biological fields. In this paper, a novel fluorescent aptasensing strategy for protein assay has been developed based on target-triggered hybridization chain reaction (HCR) and graphene oxide (GO)-based selective fluorescence quenching. Three DNA probes, a helper DNA probe (HP), hairpin probe 1 (H1) and hairpin probe 2 (H2) are ingeniously designed. In the presence of the target, the aptamer sequences in HP recognize the target to form a target-aptamer complex, which causes the HP conformation change, and then triggers the chain-like assembly of H1 and H2 through the hybridization chain reaction, generating a long chain of HP leading complex of H1 and H2. At last the fluorescence indicator SYBR Green I (SG) binds with the long double strands of the HCR product through both intercalation and minor groove binding. When GO was added into the solutions after HCR, the free H1, H2 and SG would be closely adsorbed onto GO surface via π-π stacking. However, the HCR product cannot be adsorbed on GO surface, thereby the SG bound to HCR product gives a strong fluorescence signal dependent on the concentration of the target. With the use of platelet-derived growth factor BB (PDGF-BB) as the model analyte, this newly designed protocol provides a highly sensitive fluorescence detection of PDGF-BB with a limit of detection down to 1.25 pM, and also exhibit good selectivity and applicability in complex matrixes. Therefore, the proposed aptasensing strategy based on target-triggered hybridization chain reaction amplification should have wide applications in the diagnosis of genetic diseases due to its simplicity, low cost, and high sensitivity at extremely low target concentrations.

  18. 78 FR 60302 - National Advisory Council

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-10-01

    ... SECURITY Federal Emergency Management Agency National Advisory Council AGENCY: Federal Emergency Management Agency, DHS. ACTION: Committee Management; Notice of Federal Advisory Committee Meeting. SUMMARY: The Federal Emergency Management Agency (FEMA) National Advisory Council (NAC) will meet in person on...

  19. 75 FR 56997 - Global Markets Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-09-17

    ... COMMISSION Global Markets Advisory Committee AGENCY: Commodity Futures Trading Commission (``CFTC''). ACTION: Notice of meeting of Global Markets Advisory Committee. SUMMARY: The Global Markets Advisory Committee...., Washington, DC 20581, attention Office of the Secretary. Please use the title ``Global Markets...

  20. 77 FR 43064 - Meeting; Technology Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-07-23

    ... COMMISSION Meeting; Technology Advisory Committee AGENCY: Commodity Futures Trading Commission (CFTC). ACTION: Notice of emergency meeting of technology advisory committee. SUMMARY: The CFTC announces that on Thursday, July 26, 2012, the CFTC's Technology Advisory Committee (``TAC'') will hold an emergency...

  1. 75 FR 58367 - Technology Advisory Committee Meeting

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-09-24

    ... COMMISSION Technology Advisory Committee Meeting AGENCY: Commodity Futures Trading Commission (``CFTC''). ACTION: Notice of meeting of Technology Advisory Committee. SUMMARY: The Technology Advisory Committee...., Washington, DC 20581, attention: Office of the Secretary. Please use the title ``Technology...

  2. 76 FR 776 - Technology Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-01-06

    ... COMMISSION Technology Advisory Committee AGENCY: Commodity Futures Trading Commission (``CFTC''). ACTION: Notice of meeting of Technology Advisory Committee. SUMMARY: The Technology Advisory Committee will hold...., Washington, DC 20581, attention: Office of the Secretary. Please use the title ``Technology...

  3. 78 FR 50040 - Technology Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-08-16

    ... COMMISSION Technology Advisory Committee AGENCY: Commodity Futures Trading Commission. ACTION: Notice of Meeting of Technology Advisory Committee. SUMMARY: The Commodity Futures Trading Commission (CFTC) announces that on September 12, 2013, the CFTC's Technology Advisory Committee (TAC) will hold a...

  4. 76 FR 41196 - Shoshone Resource Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-07-13

    ... Forest Service Shoshone Resource Advisory Committee AGENCY: Forest Service, USDA. ACTION: Notice of meeting. SUMMARY: The Shoshone Resource Advisory Committee (Committee) will meet in Thermopolis, Wyoming...: Olga Troxel, Resource Advisory Committee Coordinator, Shoshone National Forest Supervisor's...

  5. 76 FR 3081 - Shoshone Resource Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-01-19

    ... Forest Service Shoshone Resource Advisory Committee AGENCY: Forest Service, USDA. ACTION: Notice of meeting. SUMMARY: The Shoshone Resource Advisory Committee (Committee) will hold a conference call on.... FOR FURTHER INFORMATION CONTACT: Olga Troxel, Resource Advisory Committee Coordinator,...

  6. 77 FR 51512 - Allegheny Resource Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-08-24

    ... Forest Service Allegheny Resource Advisory Committee AGENCY: Forest Service, USDA. ACTION: Notice of meeting. SUMMARY: The Allegheny Resource Advisory Committee will meet in Warren, Pennsylvania. The... conducted: Allegheny Resource Advisory Committee members will solicit and consider project proposals...

  7. 76 FR 22672 - Shoshone Resource Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-04-22

    ... Forest Service Shoshone Resource Advisory Committee AGENCY: Forest Service, USDA. ACTION: Notice of meeting. SUMMARY: The Shoshone Resource Advisory Committee (Committee) will meet in Thermopolis, Wyoming... CONTACT: Olga Troxel, Resource Advisory Committee Coordinator, Shoshone National Forest...

  8. 76 FR 12317 - Shoshone Resource Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-03-07

    ... Forest Service Shoshone Resource Advisory Committee AGENCY: Forest Service, USDA. ACTION: Notice of meeting. SUMMARY: The Shoshone Resource Advisory Committee (Committee) will meet in Thermopolis, Wyoming... Broadway, Thermopolis, Wyoming. FOR FURTHER INFORMATION CONTACT: Olga Troxel, Resource Advisory...

  9. 76 FR 2081 - Ketchikan Resource Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-01-12

    ...; ] DEPARTMENT OF AGRICULTURE Forest Service Ketchikan Resource Advisory Committee AGENCY: Forest Service, USDA. ACTION: Notice of meeting. SUMMARY: The Ketchikan Resource Advisory Committee will meet in Ketchikan..., 3031 Tongass Avenue, Ketchikan, Alaska. Send written comments to Ketchikan Resource Advisory...

  10. 76 FR 31578 - Siskiyou Resource Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-06-01

    ... Forest Service Siskiyou Resource Advisory Committee AGENCY: Forest Service, USDA. ACTION: Notice of meeting. SUMMARY: The Siskiou Resource Advisory Committee will meet in Smith River, California. The..., OR Resource Advisory committee for Josephine, Coos and Curry Counties; review and recommend...

  11. Redox-sensitive micelles self-assembled from amphiphilic hyaluronic acid-deoxycholic acid conjugates for targeted intracellular delivery of paclitaxel.

    PubMed

    Li, Jing; Huo, Meirong; Wang, Jing; Zhou, Jianping; Mohammad, Jumah M; Zhang, Yinlong; Zhu, Qinnv; Waddad, Ayman Y; Zhang, Qiang

    2012-03-01

    A targeted intracellular delivery system of paclitaxel (PTX) was successfully developed based on redox-sensitive hyaluronic acid-deoxycholic acid (HA-ss-DOCA) conjugates. The conjugates self-assembled into nano-size micelles in aqueous media and exhibited excellent drug-loading capacities (34.1%) and entrapment efficiency (93.2%) for PTX. HA-ss-DOCA micelles were sufficiently stable at simulated normal physiologic condition but fast disassembled in the presence of 20 mm reducing agent, glutathione. In vitro drug release studies showed that the PTX-loaded HA-ss-DOCA micelles accomplished rapid drug release under reducing condition. Intracellular release of fluorescent probe nile red indicated that HA-ss-DOCA micelles provide an effective approach for rapid transport of cargo into the cytoplasm. Enhanced cytotoxicity of PTX-loaded HA-ss-DOCA micelles further confirmed that the sensitive micelles are more potent for intracellular drug delivery as compared to the insensitive control. Based on flow cytometry and confocal microscopic analyses, observations revealed that HA-ss-DOCA micelles were taken up to human breast adenocarcinoma cells (MDA-MB-231) via HA-receptor mediated endocytosis. In vivo investigation of micelles in tumor-bearing mice confirmed that HA-ss-DOCA micelles possessed much higher tumor targeting capacity than the insensitive control. These results suggest that redox-sensitive HA-ss-DOCA micelles hold great potential as targeted intracellular delivery carriers of lipophilic anticancer drugs.

  12. Nicking endonuclease-assisted recycling of target-aptamer complex for sensitive electrochemical detection of adenosine triphosphate.

    PubMed

    Hu, Tianxing; Wen, Wei; Zhang, Xiuhua; Wang, Shengfu

    2016-02-21

    An electrochemical biosensor was developed for the detection of adenosine triphosphate (ATP) based on target-induced conformation switching and nicking endonuclease (NEase)-assisted signal amplification. The electrochemical biosensor was constructed by base pairing and target recognition. After capture DNA hybridized with the gold electrode, a significant current of Methylene Blue (MB) was obtained by differential pulse voltammetry. In the presence of ATP, the hairpin DNA formed a G-quadruplex structure due to the specific recognition between hairpin DNA and ATP. Then the exposed part of the target-aptamer complex hybridized with the 3'-terminus of capture DNA to form a specific nicking site for Nb.BbvCI, which led to NEase-assisted target-aptamer complex recycling. The released target-aptamer complex hybridized with the remaining capture DNA. Nb.BbvCI-assisted target-aptamer complex recycling caused the continuous cleavage of capture DNA with MB at its 5'-terminus, resulting in release of a certain amount of DNA fragment labeled with MB. Then the current value decreased significantly. The reduced current showed a linear range from 10 nM to 1 μM with a limit of detection as low as 3.4 nM. Furthermore, the proposed strategy can be used for the detection of similar substances.

  13. National and Governmental Advisory Committees: Meetings

    EPA Pesticide Factsheets

    Meetings and Teleconference Information for the National Advisory Committee and Governmental Advisory Committee to the United States Representative to the North American Commission for Environmental Cooperation,

  14. 75 FR 26918 - Siuslaw Resource Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-05-13

    ...; ] DEPARTMENT OF AGRICULTURE Forest Service Siuslaw Resource Advisory Committee AGENCY: Forest Service, USDA. ACTION: Notice of meeting. SUMMARY: The Siuslaw Resource Advisory Committee will meet in Corvallis,...

  15. 75 FR 39910 - Dixie Resource Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-07-13

    ...; ] DEPARTMENT OF AGRICULTURE Forest Service Dixie Resource Advisory Committee AGENCY: Forest Service, USDA. ACTION: Notice of meetings. SUMMARY: The Dixie Resource Advisory Committee will meet in Cedar City,...

  16. 78 FR 22842 - Nicolet Resource Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-04-17

    ...; ] DEPARTMENT OF AGRICULTURE Forest Service Nicolet Resource Advisory Committee AGENCY: Forest Service, USDA. ACTION: Notice of meeting. SUMMARY: The Nicolet Resource Advisory Committee will meet in Crandon, WI....

  17. Citizen Advisory Committees.

    ERIC Educational Resources Information Center

    Miller, Leann R.

    This guide, describing community involvement through citizen advisory committees, is a summary of the literature on such committees. Its main concern is district committees created by school boards. Citations in the bibliography contain all points of view on committees and present many alternatives on most of the topics covered in the guide.…

  18. A Placement Advisory Test

    ERIC Educational Resources Information Center

    Hughes, Chris

    2010-01-01

    The primary method of placement at Portland CC (PCC) is the Compass Placement test. For the most part, students are placed correctly, but there are cases when students feel that they have been placed too low. In such cases we use our newly created Placement Advisory Test (PAT) to help us place them appropriately. (Contains 2 figures.)

  19. The UKCIS Advisory Service.

    ERIC Educational Resources Information Center

    Bingham, Archie

    1979-01-01

    Describes the free advisory service available to both users and potential users of chemical and biological databases in the United Kingdom and Ireland. Three specific areas are discussed in which queries about Chemical Abstracts Service (CAS) Registry Numbers have been received: isomers, replacing registry numbers, and mixed compounds. (JD)

  20. On the sensitivity decay of the cumarine targets for fast ozone measurements. Implications for the estimation of the instrumental zero and flux calculations

    NASA Astrophysics Data System (ADS)

    Finco, Angelo; Gerosa, Giacomo; Marzuoli, Riccardo

    2015-04-01

    Fast ozone concentrations measurements are necessary in order to measure ozone fluxes with the eddy covariance technique. Since the development of the first instrument early in the 90s several other instruments, all based on a chemiluminescent reaction between ozone and a cumarine target, were developed but only in 2010 Mueller et al. recognized the importance of estimating the zero (i.e. the voltage at zero ozone concentration) which depends both on instrument and target performances. In this work we will show a new methodology to estimate the zero, this new methodology avoids some problems which were unsolved by the Mueller's one. Our first assumption wais that the sensitivity of the targets decays in an exponential way rather than a linear one, as proposed by Mueller et al. (2010). This assumption was in agreement with what proposed by Ermel et al. (2013) Similarly to the Mueller's approach, the first step we performed was plotting the instrument voltage output versus the ozone concentrations, but two main differences were introduced in our methodology: first of all we compared periods in which the target received a comparable ozone dose and then the estimation of the zero is extrapolated with an exponential fit of the data rather a linear one. In this way it was possible to avoid negative zeroes which were sometimes obtained, especially in the first 24/36 hours of the target life, by applying Mueller's methodology; negative zeroes lead to an underestimation of the ozone fluxes . After estimating the zero for some sub-periods of the target life, the evolution of the zero is modeled by interpolating the zero data as a function of the ozone dose received by the target. Moreover, with this approach the zero changes continuously with no abrupt change during the target life, avoiding remarkable discontinuities in the fluxes. Comparisons between the two methodologies will be showed.

  1. A highly sensitive target-primed rolling circle amplification (TPRCA) method for fluorescent in situ hybridization detection of microRNA in tumor cells.

    PubMed

    Ge, Jia; Zhang, Liang-Liang; Liu, Si-Jia; Yu, Ru-Qin; Chu, Xia

    2014-02-04

    The ability to detect spatial and temporal microRNA (miRNA) distribution at the single-cell level is essential for understanding the biological roles of miRNAs and miRNA-associated gene regulatory networks. We report for the first time the development of a target-primed RCA (TPRCA) strategy for highly sensitive and selective in situ visualization of miRNA expression patterns at the single-cell level. This strategy uses a circular DNA as the probe for in situ hybridization (ISH) with the target miRNA molecules, and the free 3' terminus of miRNA then initiates an in situ RCA reaction to generate a long tandem repeated sequence with thousands of complementary segments. After hybridization with fluorescent detection probes, target miRNA molecules can be visualized with ultrahigh sensitivity. Because the RCA reaction can only be initiated by the free 3' end of target miRNA, the developed strategy offers the advantage over existing ISH methods in eliminating the interference from precursor miRNA or mRNA. This strategy is demonstrated to show high sensitivity and selectivity for the detection of miR-222 expression levels in human hepatoma SMMC-7721 cells and hepatocyte L02 cells. Moreover, the developed TPRCA-based ISH strategy is successfully applied to multiplexed detection using two-color fluorescent probes for two miRNAs that are differentially expressed in the two cell lines. The results reveal that the developed strategy may have great potential for in situ miRNA expression analysis for basic research and clinical diagnostics.

  2. Novel multifunctional pH-sensitive nanoparticles loaded into microbubbles as drug delivery vehicles for enhanced tumor targeting

    PubMed Central

    Lv, Yongjiu; Hao, Lan; Hu, Wenjing; Ran, Ya; Bai, Yan; Zhang, Liangke

    2016-01-01

    This study fabricated novel multifunctional pH-sensitive nanoparticles loaded into microbubbles (PNP-MB) with the combined advantages of two excellent drug delivery vehicles, namely, pH-sensitive nanoparticles and microbubbles. As an antitumor drug, resveratrol (RES) was loaded into acetylated β-cyclodextrin nanoparticles (RES-PNP). The drug-loaded nanoparticles were then encapsulated into the internal space of the microbubbles. The characterization and morphology of this vehicle were investigated through dynamic light scattering and confocal laser scanning microscopy, respectively. In vitro drug release was performed to investigate the pH sensitivity of RES-PNP. The antitumor property of RES-loaded PNP-MB (RES-PNP-MB) was also analyzed in vivo to evaluate the antitumor effect of RES-PNP-MB. Results suggested that PNP exhibited pH sensitivity, and was successfully encapsulated into the microbubbles. RES-PNP-MB exhibit effective tumor growth suppressing in vivo. Therefore, such drug delivery vehicle should be of great attention in tumor therapy. PMID:27378018

  3. miR-483-5p determines mitochondrial fission and cisplatin sensitivity in tongue squamous cell carcinoma by targeting FIS1.

    PubMed

    Fan, Song; Chen, Wei-Xiong; Lv, Xiao-Bin; Tang, Qiong-Lan; Sun, Li-Juan; Liu, Bo-Du; Zhong, Jiang-Long; Lin, Zhao-Yu; Wang, You-Yuan; Li, Qun-Xing; Yu, Xin; Zhang, Han-Qing; Li, Yi-Lin; Wen, Bin; Zhang, Zhang; Chen, Wei-Liang; Li, Jin-Song

    2015-07-01

    Mitochondria play an important role in the initiation of apoptosis. However, whether cisplatin can induce apoptosis by initiating a mitochondrial fission pathway and the mechanism underlying this effect remain poorly understood. In this study, we show that the mitochondrial fission protein FIS1 is upregulated upon cisplatin treatment in tongue squamous cell carcinoma (TSCC) cells. FIS1 knockdown can attenuate mitochondrial fission and cisplatin sensitivity. We found that FIS1 is a direct target of miR-483-5p and that miR-483-5p can inhibit mitochondrial fission and cisplatin sensitivity in vitro and in vivo. Furthermore, we found that miR-483-5p and FIS1 are significantly associated with cisplatin sensitivity and with overall survival in patients with TSCC in a retrospective analysis of multiple centers. This study revealed that a novel mitochondrial fission pathway composed of miR-483-5p and FIS1 regulates cisplatin sensitivity. The modulation of miR-483-5p and FIS1 levels may provide a new approach for increasing cisplatin sensitivity.

  4. Target-size embracing dimension for sensitive detection of viruses with various sizes and influenza virus strains.

    PubMed

    Lin, Ying-Yi; Liao, Jiunn-Der; Yang, Mei-Lin; Wu, Chao-Liang

    2012-05-15

    The focused ion beam (FIB) technique was employed to precisely fabricate hexagon-like Au nano-rods (fibAu_h) arrays as a surface enhanced Raman scattering - active substrate. A "ring diameter" (D(R)) was created by the convergence of three fibAu_h with respect to the dimension of the target viruses (D(T)), such as adenovirus (Adeno), encephalomyocarditis virus (EMCV), influenza virus (H1N1) with different sizes. Three influenza A virus strains were also compared. The results indicate that as that with a D(R)/D(T) ratio of around 1, the discrimination ability for detecting the target viruses and SERS mechanism become obvious. The enhanced lightning rod effect surrounding the seized target virus is anticipated if its size and dimension is suitably embraced within three fibAu_h. Hence the as-designed fibAu_h sample with a target-size embracing dimension provides good discrimination ability for distinguishing virus of various sizes or virus strains.

  5. Roles of uptake, biotransformation, and target site sensitivity in determining the differential toxicity of chlorpyrifos to second to fourth instar Chironomous riparius (Meigen)

    USGS Publications Warehouse

    Buchwalter, D.B.; Sandahl, J.F.; Jenkins, J.J.; Curtis, L.R.

    2004-01-01

    Early life stages of aquatic organisms tend to be more sensitive to various chemical contaminants than later life stages. This research attempted to identify the key biological factors that determined sensitivity differences among life stages of the aquatic insect Chironomous riparius. Specifically, second to fourth instar larvae were exposed in vivo to both low and high waterborne concentrations of chlorpyrifos to examine differences in accumulation rates, chlorpyrifos biotransformation, and overall sensitivity among instars. In vitro acetylcholinesterase (AChE) assays were performed with chlorpyrifos and the metabolite, chlorpyrifos-oxon, to investigate potential target site sensitivity differences among instars. Earlier instars accumulated chlorpyrifos more rapidly than later instars. There were no major differences among instars in the biotransformation rates of chlorpyrifos to the more polar metabolites, chlorpyrifos-oxon, and chlorpyridinol (TCP). Homogenate AChE activities from second to fourth instar larvae were refractory to chlorpyrifos, even at high concentrations. In contrast, homogenate AChE activities were responsive in a dose-dependent manner to chlorpyrifos-oxon. In general, it appeared that chlorpyrifos sensitivity differences among second to fourth instar C. riparius were largely determined by differences in uptake rates. In terms of AChE depression, fourth instar homogenates were more sensitive to chlorpyrifos and chlorpyrifos-oxon than earlier instars. However, basal AChE activity in fourth instar larvae was significantly higher than basal AChE activity in second to third instar larvae, which could potentially offset the apparent increased sensitivity to the oxon. ?? 2003 Elsevier B.V. All rights reserved.

  6. ATM mutations and E-cadherin expression define sensitivity to EGFR-targeted therapy in colorectal cancer.

    PubMed

    Geißler, Anna-Lena; Geißler, Miriam; Kottmann, Daniel; Lutz, Lisa; Fichter, Christiane D; Fritsch, Ralph; Weddeling, Britta; Makowiec, Frank; Werner, Martin; Lassmann, Silke

    2017-02-09

    EGFR-targeted therapy is a key treatment approach in patients with RAS wildtype metastatic colorectal cancers (CRC). Still, also RAS wildtype CRC may be resistant to EGFR-targeted therapy, with few predictive markers available for improved stratification of patients. Here, we investigated response of 7 CRC cell lines (Caco-2, DLD1, HCT116, HT29, LS174T, RKO, SW480) to Cetuximab and correlated this to NGS-based mutation profiles, EGFR promoter methylation and EGFR expression status as well as to E-cadherin expression. Moreover, tissue specimens of primary and/or recurrent tumors as well as liver and/or lung metastases of 25 CRC patients having received Cetuximab and/or Panitumumab were examined for the same molecular markers. In vitro and in situ analyses showed that EGFR promoter methylation and EGFR expression as well as the MSI and or CIMP-type status did not guide treatment responses. In fact, EGFR-targeted treatment responses were also observed in RAS exon 2 p.G13 mutated CRC cell lines or CRC cases and were further linked to PIK3CA exon 9 mutations. In contrast, non-response to EGFR-targeted treatment was associated with ATM mutations and low E-cadherin expression. Moreover, down-regulation of E-cadherin by siRNA in otherwise Cetuximab responding E-cadherin positive cells abrogated their response. Hence, we here identify ATM and E-cadherin expression as potential novel supportive predictive markers for EGFR-targeted therapy.

  7. A Sensitive Assay Using a Native Protein Substrate For Screening HIV-1 Maturation Inhibitors Targeting the Protease Cleavage Site between Matrix and Capsid

    PubMed Central

    Lee, Sook-Kyung; Cheng, Nancy; Hull-Ryde, Emily; Potempa, Marc; Schiffer, Celia A.; Janzen, William; Swanstrom, Ronald

    2013-01-01

    The matrix/capsid processing site in the HIV-1 Gag precursor is likely the most sensitive target to inhibit HIV-1 replication. We have previously shown that modest incomplete processing at the site leads to a complete loss of virion infectivity. In the current study, a sensitive assay based on fluorescence polarization is described that can monitor cleavage at the MA/CA site in the context of the folded protein substrate. The substrate, an MA/CA fusion protein, was labeled with the fluorescein-based FlAsH (Fluorescein Arsenical Hairpin) reagent which binds to a tetracysteine motif (CCGPCC) that was introduced within the N-terminal domain of CA. By limiting the size of CA and increasing the size of MA (with an N-terminal GST fusion), significant differences in polarization values were measurable as a function of HIV-1 protease cleavage. The sensitivity of the assay was tested in the presence of increasing amounts of an HIV-1 PR inhibitor, which resulted in a gradual decrease in the FP values demonstrating that the assay is sensitive discerning changes in protease processing. The high-throughput screening assay validation in 384-well plates showed that the assay is reproducible and robust with an average Z'–value of 0.79 and average coefficient of variation values less than 3%. The robustness and reproducibility of the assay was further validated using the LOPAC1280 compound library, demonstrating that the assay provides a sensitive high-throughput screening platform that can be used with large compound libraries for identifying novel maturation inhibitors targeting the MA/CA site of the HIV-1 Gag polyprotein. PMID:23763575

  8. MiR-424 and miR-27a increase TRAIL sensitivity of acute myeloid leukemia by targeting PLAG1

    PubMed Central

    Han, Xiao-yu; Ji, Min; Zhou, Ying; Zhang, A-min; Wang, Hong-chun; Ma, Dao-xin; Ji, Chun-yan

    2016-01-01

    Although microRNAs have been elaborated to participate in various physiological and pathological processes, their functions in TRAIL resistance of acute myeloid leukemia (AML) remain obscure. In this study, we detected relatively lower expression levels of miR-424&27a in TRAIL-resistant and semi-resistant AML cell lines as well as newly diagnosed patient samples. Overexpression of miR-424&27a, by targeting the 3′UTR of PLAG1, enhanced TRAIL sensitivity in AML cells. Correspondingly, knockdown of PLAG1 sensitized AML cells to TRAIL-induced apoptosis and proliferation inhibition. We further found that PLAG1 as a transcription factor could reinforce Bcl2 promoter activity, causing its upregulation at the mRNA level. Both downregulated PLAG1 and elevated expression of miR-424&27a led to Bcl2 downregulation and augmented cleavage of Caspase8, Caspase3 and PARP in the presence of TRAIL. Restoration of Bcl2 could eliminate their effects on AML TRAIL sensitization. Overall, we propose that miR-424&27a and/or PLAG1 might serve as novel therapeutic targets in AML TRAIL therapy. PMID:27013583

  9. miR-33-5p, a novel mechano-sensitive microRNA promotes osteoblast differentiation by targeting Hmga2

    PubMed Central

    Wang, Han; Sun, Zhongyang; Wang, Yixuan; Hu, Zebing; Zhou, Hua; Zhang, Lianchang; Hong, Bo; Zhang, Shu; Cao, Xinsheng

    2016-01-01

    MicroRNAs (miRNAs) interfere with the translation of specific target mRNAs and are thought to thereby regulate many cellular processes. However, the role of miRNAs in osteoblast mechanotransduction remains to be defined. In this study, we investigated the ability of a miRNA to respond to different mechanical environments and regulate mechano-induced osteoblast differentiation. First, we demonstrated that miR-33-5p expressed by osteoblasts is sensitive to multiple mechanical environments, microgravity and fluid shear stress. We then confirmed the ability of miR-33-5p to promote osteoblast differentiation. Microgravity or fluid shear stress influences osteoblast differentiation partially via miR-33-5p. Through bioinformatics analysis and a luciferase assay, we subsequently confirmed that Hmga2 is a target gene of miR-33-5p that negatively regulates osteoblast differentiation. Moreover, miR-33-5p regulates osteoblast differentiation partially via Hmga2. In summary, our findings demonstrate that miR-33-5p is a novel mechano-sensitive miRNA that can promote osteoblast differentiation and participate in the regulation of differentiation induced by changes in the mechanical environment, suggesting this miRNA as a potential target for the treatment of pathological bone loss. PMID:26980276

  10. Targeting Medicare consumers. Managed care providers can make inroads by understanding preference and cost-sensitivity issues.

    PubMed

    Stensrud, J; Sylvestre, E; Sivadas, E

    1997-01-01

    The authors' conjoint study provided valuable information on the preferences of the hugh Medicare-eligible and soon-to-be-eligible markets. Leading the list were hospitalization coverage, skilled nursing facilities, and out-of-area coverage. The task of defining choice sets was made easier and more meaningful by selecting the top six attributes for each respondent. Asking respondents to rank levels within each attribute and assessing the importance of the various levels provided a more robust estimate of consumer preferences. Using an innovative price-sensitivity method preserved the integrity of the data. The method minimized respondent fatigue and enabled the authors to gather price-sensitivity data from respondents who were not actually paying for their health services. Respondents preferred Supplemental F and Medicare products even though they placed more value on the qualities of alternative health care products. This suggests that managed care providers need to change consumer perceptions about their products.

  11. 78 FR 56231 - Government-Wide Travel Advisory Committee (GTAC); Public Advisory Committee Meeting

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-09-12

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  12. 77 FR 69869 - National Advisory Council on Alcohol Abuse and Alcoholism, National Advisory Council on Drug...

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  13. 78 FR 51191 - Government-wide Travel Advisory Committee (GTAC); Public Advisory Committee Meeting

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-08-20

    ... ADMINISTRATION Government-wide Travel Advisory Committee (GTAC); Public Advisory Committee Meeting AGENCY: Office... Government-wide Travel Advisory Committee (GTAC) (the Committee), is a Federal Advisory Committee established in accordance with the Federal Advisory Committee Act (FACA), 5 U.S.C., App 2. This ] notice...

  14. Bombs from On-High: Weaponized Stratospheric Airships for Close Air Support and Time-Sensitive-Target Missions

    DTIC Science & Technology

    2009-04-01

    battlefield. Using helium and solar power, WSAs may have hourly operational costs of under $500. SAs’ inherent stealth and resiliency make them... helium envelope. The airship will operate for years at high altitude providing tracking of air and ground targets.15 SMDC has also accomplished several...low-cost propulsion of WSAs an attractive feature. With solar power providing weeks to months of loiter time over an area, WSAs have no fossil

  15. Thermo-sensitive liposomes loaded with doxorubicin and lysine modified single-walled carbon nanotubes as tumor-targeting drug delivery system.

    PubMed

    Zhu, Xiali; Xie, Yingxia; Zhang, Yingjie; Huang, Heqing; Huang, Shengnan; Hou, Lin; Zhang, Huijuan; Li, Zhi; Shi, Jinjin; Zhang, Zhenzhong

    2014-11-01

    This report focuses on the thermo-sensitive liposomes loaded with doxorubicin and lysine-modified single-walled carbon nanotube drug delivery system, which was designed to enhance the anti-tumor effect and reduce the side effects of doxorubicin. Doxorubicin-lysine/single-walled carbon nanotube-thermo-sensitive liposomes was prepared by reverse-phase evaporation method, the mean particle size was 232.0 ± 5.6 nm, and drug entrapment efficiency was 86.5 ± 3.7%. The drug release test showed that doxorubicin released more quickly at 42℃ than at 37℃. Compared with free doxorubicin, doxorubicin-lysine/single-walled carbon nanotube-thermo-sensitive liposomes could efficiently cross the cell membranes and afford higher anti-tumor efficacy on the human hepatic carcinoma cell line (SMMC-7721) cells in vitro. For in vivo experiments, the relative tumor volumes of the sarcomaia 180-bearing mice in thermo-sensitive liposomes group and doxorubicin group were significantly smaller than those of N.S. group. Meanwhile, the combination of near-infrared laser irradiation at 808 nm significantly enhanced the tumor growth inhibition both on SMMC-7721 cells and the sarcomaia 180-bearing mice. The quality of life such as body weight, mental state, food and water intake of sarcomaia 180 tumor-bearing mice treated with doxorubicin-lysine/single-walled carbon nanotube-thermo-sensitive liposomes were much higher than those treated with doxorubicin. In conclusion, doxorubicin-lysine/single-walled carbon nanotube-thermo-sensitive liposomes combined with near-infrared laser irradiation at 808 nm may potentially provide viable clinical strategies for targeting delivery of anti-cancer drugs.

  16. Preparation and performance of a pH-sensitive cisplatin-loaded magnetic nanomedicine that targets tumor cells via folate receptor mediation

    PubMed Central

    CHEN, SHUAI-JUN; ZHANG, HONG-ZHENG; WAN, LIANG-CAI; JIANG, SHAN-SHAN; XU, YI-MING; LIU, FANG; ZHANG, TAO; MA, DONG; XIE, MIN-QIANG

    2016-01-01

    The present study aimed to prepare cisplatin (CDDP)-loaded magnetic nanoparticles (MNPs), which target folate receptors via a pH-sensitive release system (FA-PEG-NH-N=MNPs-CDDP). This is of interest for the development of intelligent drug delivery systems that target tumors of the head and neck. The chemical coprecipitation method was used to prepare ferroferric oxide MNPs. These were modified with aldehyde sodium alginate complexed with the chemotherapeutic agent, CDDP on the surface of the nanoparticles. Double hydrazine-poly(ethylene glycol; PEG) was also prepared by attaching the carboxyl group of hydrazine-folate on one side of the double hydrazine-PEG, obtaining folate-hydrazine-PEG-diazenyl. This binds the aldehyde group of sodium alginic acid on the MNP to enclose CDDP, in order that it is sequestered within the carrier. This method obtained a pH-sensitive, FA-modified CDDP-loaded MNP (FA-PEG-NH-N=MNPs-CDDP), which acts as an intelligent tumor targeting drug delivery system. The mean size of the MNPs was ~10.2±1.5 nm, the mean hydrodynamic diameter detected by laser particle sizing instruments was 176.6±1.1 nm, and the ζ-potential was −20.91±1.76 mV. The CDDP content was 0.773 mg/ml, the iron content was ~1.908 mg/ml and the maximum saturation magnetization was 16.3±0.2 emu/g. The current study produced a pH-sensitive FA-modified CDDP-loaded MNP that is stable and exhibits magnetic responsiveness, which releases CDDP in a low pH environment. PMID:27109546

  17. 75 FR 43156 - Federal Advisory Committee; Missile Defense Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-07-23

    ... staff and Program Managers on the Agency's strategic perspective and the Ballistic Missile Defense..., but are not limited to briefings on the Ballistic Missile Defense Review, Early Intercept, Phased... Office of the Secretary Federal Advisory Committee; Missile Defense Advisory Committee AGENCY:...

  18. Effect of Fibroblast-Like Cells of Mesenchymal Origin of Cytotoxic Activity of Lymphocytes against NK-Sensitive Target Cells.

    PubMed

    Lupatov, A Yu; Kim, Ya S; Bystrykh, O A; Vakhrushev, I V; Pavlovich, S V; Yarygin, K N; Sukhikh, G T

    2017-02-01

    We studied immunosuppressive properties of skin fibroblasts and mesenchymal stromal cells against NK cells. In vitro experiments showed that mesenchymal stromal cells isolated from human umbilical cord and human skin fibroblasts can considerably attenuate cytotoxic activity of NK cells against Jurkat cells sensitive to NK-mediated lysis. NK cells cultured in lymphocyte population exhibited higher cytotoxic activity than isolated NK cells. Mesenchymal stromal cells or fibroblasts added 1:1 to lymphocyte culture almost completely suppressed NK cell cytotoxicity. This suggests that fibroblast-like cells can suppress not only isolated NK cells, but also NK cells in natural cell microenvironment.

  19. Dual Targeting of Akt and mTORC1 Impairs Repair of DNA Double-Strand Breaks and Increases Radiation Sensitivity of Human Tumor Cells

    PubMed Central

    Holler, Marina; Grottke, Astrid; Mueck, Katharina; Manes, Julia; Jücker, Manfred

    2016-01-01

    Inhibition of mammalian target of rapamycin-complex 1 (mTORC1) induces activation of Akt. Because Akt activity mediates the repair of ionizing radiation-induced DNA double-strand breaks (DNA-DSBs) and consequently the radioresistance of solid tumors, we investigated whether dual targeting of mTORC1 and Akt impairs DNA-DSB repair and induces radiosensitization. Combining mTORC1 inhibitor rapamycin with ionizing radiation in human non-small cell lung cancer (NSCLC) cells (H661, H460, SK-MES-1, HTB-182, A549) and in the breast cancer cell line MDA-MB-231 resulted in radiosensitization of H661 and H460 cells (responders), whereas only a very slight effect was observed in A549 cells, and no effect was observed in SK-MES-1, HTB-182 or MDA-MB-231 cells (non-responders). In responder cells, rapamycin treatment did not activate Akt1 phosphorylation, whereas in non-responders, rapamycin mediated PI3K-dependent Akt activity. Molecular targeting of Akt by Akt inhibitor MK2206 or knockdown of Akt1 led to a rapamycin-induced radiosensitization of non-responder cells. Compared to the single targeting of Akt, the dual targeting of mTORC1 and Akt1 markedly enhanced the frequency of residual DNA-DSBs by inhibiting the non-homologous end joining repair pathway and increased radiation sensitivity. Together, lack of radiosensitization induced by rapamycin was associated with rapamycin-mediated Akt1 activation. Thus, dual targeting of mTORC1 and Akt1 inhibits repair of DNA-DSB leading to radiosensitization of solid tumor cells. PMID:27137757

  20. miR-337-3p and Its Targets STAT3 and RAP1A Modulate Taxane Sensitivity in Non-Small Cell Lung Cancers

    PubMed Central

    Du, Liqin; Subauste, Maria C.; DeSevo, Christopher; Zhao, Zhenze; Baker, Michael; Borkowski, Robert; Schageman, Jeoffrey J.; Greer, Rachel; Yang, Chin-Rang; Suraokar, Milind; Wistuba, Ignacio I.; Gazdar, Adi F.; Minna, John D.; Pertsemlidis, Alexander

    2012-01-01

    NSCLC (non-small cell lung cancer) often exhibits resistance to paclitaxel treatment. Identifying the elements regulating paclitaxel response will advance efforts to overcome such resistance in NSCLC therapy. Using in vitro approaches, we demonstrated that over-expression of the microRNA miR-337-3p sensitizes NCI-H1155 cells to paclitaxel, and that miR-337-3p mimic has a general effect on paclitaxel response in NSCLC cell lines, which may provide a novel adjuvant strategy to paclitaxel in the treatment of lung cancer. By combining in vitro and in silico approaches, we identified STAT3 and RAP1A as direct targets that mediate the effect of miR-337-3p on paclitaxel sensitivity. Further investigation showed that miR-337-3p mimic also sensitizes cells to docetaxel, another member of the taxane family, and that STAT3 levels are significantly correlated with taxane resistance in lung cancer cell lines, suggesting that endogenous STAT3 expression is a determinant of intrinsic taxane resistance in lung cancer. The identification of a miR-337-3p as a modulator of cellular response to taxanes, and STAT3 and RAP1A as regulatory targets which mediate that response, defines a novel regulatory pathway modulating paclitaxel sensitivity in lung cancer cells, which may provide novel adjuvant strategies along with paclitaxel in the treatment of lung cancer and may also provide biomarkers for predicting paclitaxel response in NSCLC. PMID:22723956

  1. Quantum dots-labeled strip biosensor for rapid and sensitive detection of microRNA based on target-recycled nonenzymatic amplification strategy.

    PubMed

    Deng, Huaping; Liu, Qianwen; Wang, Xin; Huang, Ru; Liu, Hongxing; Lin, Qiumei; Zhou, Xiaoming; Xing, Da

    2017-01-15

    MicroRNAs (miRNAs) have been proved to be potential biomarkers in early cancer diagnosis. It is of great significance for rapid and sensitive detection of miRNAs, particularly with point-of-care (POC) diagnosis. Herein, it is the first time to construct quantum dots (QDs)-labeled strip biosensor based on target-recycled nonenzymatic amplification strategy for miRNA detection. In the system, QDs were served as bright, photostable signal labels, which endow this biosensor with good detection efficiency. Moreover, a target-recycled amplification strategy relies on sequence-specific hairpins strand displacement process without the assistance of enzymes, was introduced to further improve the sensitivity. Meanwhile eliminating the requirement of environment-susceptible enzyme protein makes it easy to preserve and enhances the stability and reproducibility of this sensor. Benefiting from these outstanding characteristics, this platform exhibited a good detection sensitivity range from 2fmol to 200fmol with a limit of 200amol, using only 20μL of sample within 80min. The assay was also 10-fold more sensitive than that with a conventional colloidal gold-based test strip for miRNA detection. Additionally, the analysis of miRNA in various tumor cell extracts was in accordance with the performance of quantitative realtime polymerase chain reaction (qRT-PCR). Clinical tumor samples were also tested, and 16 of 20 samples gave out positive signals, which demonstrated the practical application capacity of the biosensor. Therefore, the proposed biosensor holds great promise for potential POC applications and early cancer diagnosis.

  2. Highly sensitive detection of 25-HydroxyvitaminD3 by using a target-induced displacement of aptamer.

    PubMed

    Lee, Bang Hyun; Nguyen, Van Thuan; Gu, Man Bock

    2017-02-15

    For the prevention of 25-HydroxyvitaminD3 deficiency, in this study, aptamers which can bind to 25-HydroxyvitaminD3 with high specificity and affinity, were successfully developed by using immobilization-free, graphene oxide-based systemic evolution of ligands by exponential enrichment (GO-SELEX) method. The 9 sequences including VDBA14 aptamer were obtained out of 16 aptamer candidates, based on the specificity and affinity of the aptamers confirmed by both the gold nanoparticles (AuNPs)-based colorimetric assay and the isothermal titration calorimetry (ITC) method. Among them, the aptamer, VDBA14, developed in this study was found to show a great affinity to 25-HydroxyvitaminD3, with 11nM of its Kd value. Moreover, the circular dichroism (CD) analysis data indicated the target-induced displacement of the aptamer VDBA14clearly. In addition, this target-induced change of the aptamer was also confirmed again by conducting two different experimental formats, the use of streptavidin-coated 96-well plates and the use of magnetic beads. The results clearly indicated that the structure of VDBA14 aptamer was changed upon the binding of the target, 25-HydroxyvitaminD3, and so the indicator sequences (partially complementary to the aptamer sequence) tagged with an enzyme as a signaling molecule could be de-hybridized from the aptamer. Finally, the limit of detection for vitamin D based on AuNPs-based colorimetric assay using VDBA14 aptamer was found to be 1µM. All these results were taken together, the aptamer which was developed could play an exquisite role in the fields of early medical diagnosis of vitamin D deficiency with accurate, rapid and simple analytical method.

  3. 77 FR 55218 - Homeland Security Advisory Council

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  4. 76 FR 55079 - Homeland Security Advisory Council

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  8. 77 FR 64532 - Homeland Security Advisory Council

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  9. 75 FR 59278 - Homeland Security Advisory Council

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-09-27

    ... SECURITY Homeland Security Advisory Council AGENCY: The Office of Policy, DHS. ACTION: Notice of Closed Federal Advisory Committee Meeting. SUMMARY: The Homeland Security Advisory Council (HSAC) will meet on... message. Fax: (202) 282-9207. Mail: Homeland Security Advisory Council, Department of Homeland...

  10. 77 FR 59627 - Homeland Security Advisory Council

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-09-28

    ... SECURITY Homeland Security Advisory Council AGENCY: The Office of Policy, DHS. ACTION: Notice of open teleconference federal advisory committee meeting. SUMMARY: The Homeland Security Advisory Council (HSAC) will... line of the message. Fax: (202) 282-9207. Mail: Homeland Security Advisory Council, Department...

  11. 78 FR 70317 - Invasive Species Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-11-25

    ... Office of the Secretary Invasive Species Advisory Committee AGENCY: Office of the Secretary, Interior. ACTION: Notice of Public Meeting (via Teleconference) of the Invasive Species Advisory Committee. SUMMARY... Invasive Species Advisory Committee. The purpose of the Advisory Committee is to provide advice to...

  12. 78 FR 77443 - Electricity Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-12-23

    ... Electricity Advisory Committee AGENCY: Office of Electricity Delivery and Energy Reliability, Department of... Electricity Advisory Committee (EAC). The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770.../oe/services/electricity-advisory-committee-eac . FOR FURTHER INFORMATION CONTACT: Matthew...

  13. 16 CFR 1018.26 - Advisory functions.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 16 Commercial Practices 2 2010-01-01 2010-01-01 false Advisory functions. 1018.26 Section 1018.26 Commercial Practices CONSUMER PRODUCT SAFETY COMMISSION GENERAL ADVISORY COMMITTEE MANAGEMENT Operation of Advisory Committees § 1018.26 Advisory functions. (a) Unless otherwise specifically provided by...

  14. Targeted in vivo delivery of EGFR siRNA inhibits ovarian cancer growth and enhances drug sensitivity

    PubMed Central

    Satpathy, Minati; Mezencev, Roman; Wang, Lijuan; McDonald, John F.

    2016-01-01

    A functionalized nanohydrogel siRNA delivery system and a mouse model of serous ovarian cancer were used to test predictions from previous cell line studies that knockdown of EGFR (epidermal growth factor receptor) may be of clinical significance in the treatment of epithelial tumors especially with respect to the enhancement of platinum based therapies. Our results support these predictions and suggest that targeted delivery of EGFR siRNA may be an effective strategy for the treatment of ovarian and other epithelial tumors associated with elevated levels of EGFR and especially those demonstrating resistance to platinum-based therapies. PMID:27819259

  15. Target loads of atmospheric sulfur deposition for the protection and recovery of acid-sensitive streams in the Southern Blue Ridge Province.

    PubMed

    Sullivan, Timothy J; Cosby, Bernard J; Jackson, William A

    2011-11-01

    An important tool in the evaluation of acidification damage to aquatic and terrestrial ecosystems is the critical load (CL), which represents the steady-state level of acidic deposition below which ecological damage would not be expected to occur, according to current scientific understanding. A deposition load intended to be protective of a specified resource condition at a particular point in time is generally called a target load (TL). The CL or TL for protection of aquatic biota is generally based on maintaining surface water acid neutralizing capacity (ANC) at an acceptable level. This study included calibration and application of the watershed model MAGIC (Model of Acidification of Groundwater in Catchments) to estimate the target sulfur (S) deposition load for the protection of aquatic resources at several future points in time in 66 generally acid-sensitive watersheds in the southern Blue Ridge province of North Carolina and two adjoining states. Potential future change in nitrogen leaching is not considered. Estimated TLs for S deposition ranged from zero (ecological objective not attainable by the specified point in time) to values many times greater than current S deposition depending on the selected site, ANC endpoint, and evaluation year. For some sites, one or more of the selected target ANC critical levels (0, 20, 50, 100μeq/L) could not be achieved by the year 2100 even if S deposition was reduced to zero and maintained at that level throughout the simulation. Many of these highly sensitive streams were simulated by the model to have had preindustrial ANC below some of these target values. For other sites, the watershed soils contained sufficiently large buffering capacity that even very high sustained levels of atmospheric S deposition would not reduce stream ANC below common damage thresholds.

  16. miR-181b increases drug sensitivity in acute myeloid leukemia via targeting HMGB1 and Mcl-1.

    PubMed

    Lu, Fei; Zhang, Jingru; Ji, Min; Li, Peng; Du, Yahui; Wang, Hongchun; Zang, Shaolei; Ma, Daoxin; Sun, Xiulian; Ji, Chunyan

    2014-07-01

    Multidrug resistance (MDR) remains the major cause of disease relapse and poor prognosis in adults with acute myeloid leukemia (AML). Emerging evidence shows that drug resistance not only exists against conventional chemotherapeutic drugs, but also limits the efficacy of new biological agents. Therefore, it is important to elucidate the mechanisms through which AML patients develop drug resistance. MicroRNAs have been shown to play an important role in regulating the chemotherapy resistance in AML. A detailed understanding of the mechanisms of microRNA that are clinically relevant in AML may enhance our ability to predict and overcome drug resistance. Here, we demonstrated, for the first time, that miR-181b was decreased significantly in human multidrug-resistant leukemia cells and relapsed/refractory AML patient samples. Overexpression of miR-181b increased the sensitivity of leukemia cells to cytotoxic chemotherapeutic agents and promoted drug-induced apoptosis. Moreover, miR-181b inhibited HMGB1 and Mcl-1 expression by direct binding to their 3'-untranslated regions. In addition, HMGB1 was expressed at high levels in relapsed/refractory AML patients and suppression of HMGB1 via RNA interference sensitized multidrug-resistant leukemia cells to chemotherapy and induced apoptosis. In conclusion, these results provide a strong rationale for the development of miR-181b-based therapeutic strategies for the enhancement of efficacy in AML treatment.

  17. Ash cloud aviation advisories

    SciTech Connect

    Sullivan, T.J.; Ellis, J.S.; Schalk, W.W.; Nasstrom, J.S.

    1992-06-25

    During the recent (12--22 June 1991) Mount Pinatubo volcano eruptions, the US Air Force Global Weather Central (AFGWC) requested assistance of the US Department of Energy`s Atmospheric Release Advisory Capability (ARAC) in creating volcanic ash cloud aviation advisories for the region of the Philippine Islands. Through application of its three-dimensional material transport and diffusion models using AFGWC meteorological analysis and forecast wind fields ARAC developed extensive analysis and 12-hourly forecast ash cloud position advisories extending to 48 hours for a period of five days. The advisories consisted of ``relative`` ash cloud concentrations in ten layers (surface-5,000 feet, 5,000--10,000 feet and every 10,000 feet to 90,000 feet). The ash was represented as a log-normal size distribution of 10--200 {mu}m diameter solid particles. Size-dependent ``ashfall`` was simulated over time as the eruption clouds dispersed. Except for an internal experimental attempt to model one of the Mount Redoubt, Alaska, eruptions (12/89), ARAC had no prior experience in modeling volcanic eruption ash hazards. For the cataclysmic eruption of 15--16 June, the complex three-dimensional atmospheric structure of the region produced dramatically divergent ash cloud patterns. The large eruptions (> 7--10 km) produced ash plume clouds with strong westward transport over the South China Sea, Southeast Asia, India and beyond. The low-level eruptions (< 7 km) and quasi-steady-state venting produced a plume which generally dispersed to the north and east throughout the support period. Modeling the sequence of eruptions presented a unique challenge. Although the initial approach proved viable, further refinement is necessary and possible. A distinct need exists to quantify eruptions consistently such that ``relative`` ash concentrations relate to specific aviation hazard categories.

  18. Aerospace safety advisory panel

    NASA Technical Reports Server (NTRS)

    1994-01-01

    This report from the Aerospace Safety Advisory Panel (ASAP) contains findings, recommendations, and supporting material concerning safety issues with the space station program, the space shuttle program, aeronautics research, and other NASA programs. Section two presents findings and recommendations, section three presents supporting information, and appendices contain data about the panel membership, the NASA response to the March 1993 ASAP report, and a chronology of the panel's activities during the past year.

  19. Interactions of calmodulin with metal ions and with its target proteins revealed by conformation-sensitive monoclonal antibodies.

    PubMed

    Wolf, T; Solomon, B; Ivnitski, D; Rishpon, J; Fleminger, G

    1998-01-01

    Two monoclonal antibodies (mAbs) raised against bovine calmodulin (CaM), CAM1 and CAM4, enable one to monitor conformational changes that occur in the molecule. The interaction of CAM1 with CaM depends on the Ca2+ occupancy of its Ca(2+)-binding sites. CAM4, in contrast, interacts with CaM in a Ca(2+)-independent manner, interacting with both holoCaM and EGTA-treated CaM to a similar extent. Their interaction with various CaMs, CaM tryptic fragments and chemically modified CaM, as well as molecular graphics, led to identification of the CAM1 and CAM4 epitopes on the C- and N-terminal lobes of CAM respectively. The two mAbs were used as macromolecular probes to detect conformational changes occurring in the CaM molecule upon binding of metal ions and target proteins and peptides. MAb CAM1 successfully detected changes associated with Al3+ binding even in the presence of Ca2+, indicating that Al3+ and Ca2+ ions may bind to the protein simultaneously, leading to a new conformation of the molecule. MAbs CAM1 and CAM4 were used to follow the interactions of CaM with its target peptides and proteins. Complexes with melittin, mastoparan, calcineurin and phosphodiesterase showed different immunological properties on an immuno-enzyme electrode, indicating unique structural properties for each complex.

  20. A Silver(I)-Estrogen Nanocluster: GSH Sensitivity and Targeting Suppression on HepG2 Cell.

    PubMed

    Shen, De-Feng; Wu, Shan-Shan; Wang, Rui-Rui; Zhang, Qiang; Ren, Zhong-Juan; Liu, Hong; Guo, Hua-Dong; Gao, Guang-Gang

    2016-11-01

    A structure-determined silver nanocluster of [Ag10 (Eth)4 (CF3 COO)6 (CH3 OH)3 ]·3C-H3 OH (Eth = ethisterone) (1), is firstly demonstrated by self-assembly of silver salt and ethisterone. Due to the thiophilicity of silver(I) ions, complex 1 shows reactivity with glutathione (GSH) molecules in solution and induces the fluorescence quenching behavior. Thus, complex 1 can be used as a fluorescent sensor for GSH. In consideration of the higher level of GSH in cancerous cells, complex 1 presents significant tumor suppression reactivity toward the human hepatocellular carcinoma (HepG2) cells with IC50 value of 165 × 10(-9) m. Especially, complex 1 displays 3.4-fold higher in vitro cytotoxicity to HepG2 cells than that of the normal CCC-HEL-1 cells, which makes complex 1 a potential targeting suppression agent for cancerous cells. The molecular design of complex 1 not only generates a new medicine-silver(I) cluster family, but also opens a new avenue to the targeting anticancer organosilver(I) materials.

  1. In-Depth Study of the Interaction, Sensitivity, and Gating Modulation by PUFAs on K+ Channels; Interaction and New Targets

    PubMed Central

    Moreno, Cristina; de la Cruz, Alicia; Valenzuela, Carmen

    2016-01-01

    Voltage gated potassium channels (KV) are membrane proteins that allow selective flow of K+ ions in a voltage-dependent manner. These channels play an important role in several excitable cells as neurons, cardiomyocytes, and vascular smooth muscle. Over the last 20 years, it has been shown that omega-3 polyunsaturated fatty acids (PUFAs) enhance or decrease the activity of several cardiac KV channels. PUFAs-dependent modulation of potassium ion channels has been reported to be cardioprotective. However, the precise cellular mechanism underlying the cardiovascular benefits remained unclear in part because new PUFAs targets and signaling pathways continue being discovered. In this review, we will focus on recent data available concerning the following aspects of the KV channel modulation by PUFAs: (i) the exact residues involved in PUFAs-KV channels interaction; (ii) the structural PUFAs determinants important for their effects on KV channels; (iii) the mechanism of the gating modulation of KV channels and, finally, (iv) the PUFAs modulation of a few new targets present in smooth muscle cells (SMC), KCa1.1, K2P, and KATP channels, involved in vascular relaxation. PMID:27933000

  2. Sensitivity of KRAS-Mutant Colorectal Cancers to Combination Therapy that Co-Targets MEK and CDK4/6

    PubMed Central

    Ziemke, Elizabeth K.; Dosch, Joseph S.; Maust, Joel D.; Shettigar, Amrith; Sen, Ananda; Welling, Theodore H.; Hardiman, Karin M.; Sebolt-Leopold, Judith S.

    2015-01-01

    Purpose The emerging need for rational combination treatment approaches led us to test the concept that co-targeting MEK and CDK4/6 would prove efficacious in KRAS mutant (KRASmt) colorectal cancers, where upregulated CDK4 and hyperphosphorylated retinoblastoma (RB) typify the vast majority of tumors. Experimental Design Initial testing was carried out in the HCT-116 tumor model, which is known to harbor a KRAS mutation. Efficacy studies were then performed with five RB+ patient-derived colorectal xenograft models, genomically diverse with respect to KRAS, BRAF, and PIK3CA mutational status. Tolerance, efficacy, and pharmacodynamic evaluation of target modulation were evaluated in response to daily dosing with either agent alone or concurrent co-administration. Results Synergy was observed in vitro when HCT-116 cells were treated over a broad range of doses of trametinib and palbociclib. Subsequent in vivo evaluation of this model showed a higher degree of antitumor activity resulting from the combination compared to that achievable with single agent treatment. Testing of colorectal patient-derived xenograft (PDX) models further showed that combination of trametinib and palbociclib was well tolerated and resulted in objective responses in all KRASmt models tested. Stasis was observed in a KRAS/BRAF wild type and a BRAFmt model. Conclusions Combination of trametinib and palbociclib was well tolerated and highly efficacious in all three KRAS mutant CRC PDX models tested. Promising preclinical activity seen here supports clinical evaluation of this treatment approach to improve therapeutic outcome for metastatic colorectal cancer patients. PMID:26369631

  3. 77 FR 57085 - Federal Advisory Committee Act; Advisory Committee on Diversity for Communications in the Digital...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-09-17

    ... COMMISSION Federal Advisory Committee Act; Advisory Committee on Diversity for Communications in the Digital... Communications Commission's (FCC) Advisory Committee on Diversity for Communications in the Digital Age (``Diversity Committee''). The Committee's mission is to provide recommendations to the Commission...

  4. 76 FR 64348 - Federal Advisory Committee Act; Advisory Committee on Diversity for Communications in the Digital...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-10-18

    ... COMMISSION Federal Advisory Committee Act; Advisory Committee on Diversity for Communications in the Digital... Communications Commission's (FCC) Advisory Committee on Diversity for Communications in the Digital Age (``Diversity Committee''). The Committee's mission is to provide recommendations to the Commission...

  5. 78 FR 21354 - Federal Advisory Committee Act; Advisory Committee on Diversity for Communications in the Digital...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-04-10

    ... COMMISSION Federal Advisory Committee Act; Advisory Committee on Diversity for Communications in the Digital... Communications Commission's (FCC) Advisory Committee on Diversity for Communications in the Digital Age (``Diversity Committee''). The Committee's mission is to provide recommendations to the Commission...

  6. 77 FR 6113 - Federal Advisory Committee Act; Advisory Committee on Diversity for Communications in the Digital...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-02-07

    ... COMMISSION Federal Advisory Committee Act; Advisory Committee on Diversity for Communications in the Digital... Communications Commission's (FCC's) Advisory Committee on Diversity for Communications in the Digital Age (``Diversity Committee''). The Committee's mission is to provide recommendations to the Commission...

  7. 77 FR 55863 - NASA Advisory Council; Science Committee; Earth Science Subcommittee; Applied Sciences Advisory...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-09-11

    ... SPACE ADMINISTRATION NASA Advisory Council; Science Committee; Earth Science Subcommittee; Applied Sciences Advisory Group Meeting AGENCY: National Aeronautics and Space Administration. ACTION: Notice of... Aeronautics and Space Administration (NASA) announces a meeting of the Applied Science Advisory Group....

  8. 78 FR 64941 - Government-Wide Travel Advisory Committee (GTAC); Public Advisory Committee Meetings

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-10-30

    ... ADMINISTRATION Government-Wide Travel Advisory Committee (GTAC); Public Advisory Committee Meetings AGENCY..., Designated Federal Officer (DFO), Government-wide Travel ] Advisory Committee (GTAC), Office of Government... recommendations to existing travel policies, processes and procedures, including the per diem methodology...

  9. 75 FR 4069 - Science Advisory Board Staff Office; Notification of a Clean Air Scientific Advisory Committee...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-01-26

    ... Clean Air Scientific Advisory Committee (CASAC) Ambient Air Monitoring & Methods Subcommittee (AAMMS or... AGENCY Science Advisory Board Staff Office; Notification of a Clean Air Scientific Advisory Committee (CASAC); Ambient Air Methods and Monitoring Subcommittee (AAMMS); Meeting and Public...

  10. 76 FR 70425 - Federal Advisory Committee; Defense Intelligence Agency (DIA) Advisory Board; Closed Meeting

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-11-14

    ... Office of the Secretary Federal Advisory Committee; Defense Intelligence Agency (DIA) Advisory Board... discussions of classified information relating to DIA's intelligence operations including its support to... Advisory Board to discuss DIA operations and capabilities in support of current intelligence...

  11. 77 FR 2277 - Federal Advisory Committee; Defense Intelligence Agency (DIA) Advisory Board; Closed Meeting

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-01-17

    ... of the Secretary Federal Advisory Committee; Defense Intelligence Agency (DIA) Advisory Board; Closed... discussions of classified information relating to DIA's intelligence operations including its support to... Advisory Board to discuss DIA operations and capabilities in support of current intelligence...

  12. 77 FR 70482 - Notice of Establishment of a NASA Federal Advisory Committee; Applied Sciences Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-11-26

    ... SPACE ADMINISTRATION Notice of Establishment of a NASA Federal Advisory Committee; Applied Sciences... the establishment of the Applied Sciences Advisory Committee as a Federal advisory committee under... Integration and Management Division, Science Mission Directorate, NASA Headquarters, 300 E Street...

  13. Federal Advisory Committees: An Overview

    DTIC Science & Technology

    2009-04-16

    initiating a tradition of presidential use of outside expertise when, in 1794 , he appointed an ad hoc group of commissioners to investigate the Whiskey ... Rebellion .7 Since 1842, Congress has legislated control over federal advisory bodies — mostly by limiting funding and committee member pay. In 1842...reports aggregated advisory body information to Congress annually.12 7 " Whiskey Insurrection in

  14. OCLC and Its Advisory Committees.

    ERIC Educational Resources Information Center

    Baker, Shirley K.

    1998-01-01

    Describes the Online Computer Library Center (OCLC) advisory committees in terms of research, public, college and university, and special libraries. All four of the type-of-library advisory groups work to shape OCLC policies and programs, according to the particular needs of each group. OCLC's financial and programmatic success depends upon…

  15. Stepwise targeted drug delivery to liver cancer cells for enhanced therapeutic efficacy by galactose-grafted, ultra-pH-sensitive micelles.

    PubMed

    Yan, Guoqing; Wang, Jun; Hu, Liefeng; Wang, Xin; Yang, Guanqing; Fu, Shengxiang; Cheng, Xu; Zhang, Panpan; Tang, Rupei

    2017-03-15

    To promote drug accumulation and cell-killing ability at tumor tissue, we have prepared a stepwise targeted drug delivery system that can remain stealthy and long-circulating in the blood vessels, improve drug retention at extracellular stimuli, enhance cellular uptake through special targeting ligands, and then achieve rapid drug release to improve toxicity to tumor cells at intracellular stimuli. Herein, galactose-grafted, ultra-pH-sensitive drug carriers (POEAd-g-LA-DOX micelles), which could respond to both extracellular and intracellular pH, and combine with galactose-receptors in cell membrane, were constructed by a facile method, therefore achieving: (i) remaining stable at pH 7.4; (ii) responding to tumoral extracellular pH following gradually larger nanoparticles (NPs); (iii) conjugating receptors in the cell membrane of liver cancer through surface galactose-ligands of micelles; (iv) being sensitive to tumoral intracellular pH following further swelling for rapid drug release. In vitro cytotoxicity and cellular uptake measurement showed that POEAd-g-LA20-DOX micelle was more easily internalized and more toxic effect on tumor cells than free DOX. Moreover, in vivo biodistribution and tumor inhibition examinations demonstrated that POEAd-g-LA20-DOX formulation had more superior efficacy to significantly enhance drug accumulation in tumor, and then restrain tumor growth while decreasing drug concentration in heart.

  16. Microwave-accelerated surface plasmon-coupled directional luminescence 2: a platform technology for ultra fast and sensitive target DNA detection in whole blood.

    PubMed

    Aslan, Kadir; Previte, Michael J R; Zhang, Yongxia; Geddes, Chris D

    2008-02-29

    The application of Microwave-Accelerated Surface Plasmon-Coupled Luminescence (MA-SPCL) to fast and sensitive DNA hybridization assays in buffer and whole blood is presented. In this regard, a model DNA hybridization assay whereby a fluorophore-labeled target ssDNA specific to human immunodeficiency, Hepatitis C (Hep C), is probed by an anchor probe immobilized on thin gold films, is driven to completion within 1 min with microwave heating, as compared to an identical assay completed in approximately 4 h at room temperature. Finite-Difference Time-Domain calculations show that gold disks are preferentially heated around the edges creating a temperature gradient along the disks, which in turn results in the larger influx of complementary DNA towards anchor probe-modified surface. Thermal images of the assay platform during microwave heating also provide additional information on the microwave heating pattern in the microwave cavity. Finally, the effects of low power microwave heating on the ability of DNA to re-hybridize with the complimentary target on the surface gold films, which allows the multiple re-use of the gold films, is demonstrated. The MA-SPCL technique offers an alternative approach to current DNA based detection technologies, especially when speed and sensitivity are required, such as in the identification of DNA or even RNA-based diseases using whole blood samples that affect human health.

  17. Gastrointestinal syndrome and mucosal clonogenic cells: relationships between target cell sensitivities, LD/sub 50/ and cell survival, and their modification by antibiotics

    SciTech Connect

    Hendry, J.H.; Potten, C.S.; Roberts, N.P.

    1983-10-01

    The sensitivity of the target cells responsible for the gastrointestinal syndrome in mice was deduced from the steepness of the dose-survival curve for mice assessed on Day 7 after irradiation. The D/sub 0/ value was 1.25 +- 0.22 Gy, virtually identical to the value of 1.23 +- 0.08 measured for microcolony-forming cells (clonogens) over about the same range of dose in concurrent experiments. The survival of clonogens was similar when assayed in mice surviving to Days 3, 4, or 5, but clonogenic sensitivity was lower when assessed on Day 7. This was shown at one dose to be due largely to a selection of mice with high colony counts with only a small contribution from crypt budding. The LD/sub 50/ for mice corresponded to a surviving fraction of crypts of about 0.35. An injection of 5 mg streptomycin sulphate ip daily for 5 days after irradition increased the latent period by about 1 day, increased the LD/sub 50/ by about 1.4 Gy, but did not significantly change the survival of clonogens. These studies are the first to test and satisfy the interpretation of a dose-response curve for animal survival in terms of target cell survival, where measurements of both are made over a similar range of dose in concurrent experiments.

  18. 77 FR 62312 - Debt Management Advisory Committee; Meeting

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-10-12

    ... No: 2012-24947] DEPARTMENT OF THE TREASURY Debt Management Advisory Committee; Meeting Notice is... following debt management advisory committee: Treasury Borrowing Advisory Committee of The Securities... operations. Historically, this advice has been offered by debt management advisory committees established...

  19. 76 FR 7551 - Advisory Committee on Student Financial Assistance: Hearing

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-02-10

    ... Advisory Committee on Student Financial Assistance: Hearing AGENCY: Advisory Committee on Student Financial... Student Financial Assistance (the Advisory Committee). This notice also describes the functions of the.... Alison Bane, Associate Director of Government Relations, Advisory Committee on Student...

  20. mTOR is a Promising Therapeutic Target Both in Cisplatin-Sensitive and Cisplatin-Resistant Clear Cell Carcinoma of the Ovary

    PubMed Central

    Mabuchi, Seiji; Kawase, Chiaki; Altomare, Deborah A.; Morishige, Kenichirou; Sawada, Kenjiro; Hayashi, Masami; Tsujimoto, Masahiko; Yamoto, Mareo; Klein-Szanto, Andres J.; Schilder, Russell J.; Ohmichi, Masahide; Testa, Joseph R.; Kimura, Tadashi

    2009-01-01

    Translational Relevance Clear cell carcinoma (CCC) of the ovary is a distinctive subtype of epithelial ovarian cancer associated with a poorer sensitivity to platinum-based chemotherapy and a worse prognosis than the more common serous adenocarcinoma (SAC). To improve survival, the development of new treatment strategies that target CCC more effectively is necessary. Our results show that mTOR is more frequently activated in CCCs than in SACs. Our data have relevance for the design of future clinical studies of first-line treatment for patients with CCC of the ovary. Moreover, the finding of increased expression of phospho-mTOR and greater sensitivity to RAD001 in cisplatin-resistant CCC cells than in cisplatin-sensitive cells suggests a novel treatment option for patients with recurrent disease after cisplatin-based first-line chemotherapy. Purpose mTOR (mammalian target of rapamycin) plays a central role in cell proliferation and is regarded as a promising target in cancer therapy including for ovarian cancer. This study aims to examine the role of mTOR as a therapeutic target in clear cell carcinoma (CCC) of the ovary which is regarded as aggressive, chemo-resistant histological subtype. Experimental Design Using tissue microarrays of 98 primary ovarian cancers (52 clear cell carcinomas and 46 serous adenocarcinomas), the expression of phospho-mTOR was assessed by immunohistochemistry. Then, the growth-inhibitory effect of mTOR inhibition by RAD001 (everolimus) was examined using 2 pairs of cisplatin-sensitive parental (RMG1 and KOC7C) and cisplatin-resistant human CCC cell lines (RMG1-CR and KOC7C-CR) both in vitro and in vivo. Results Immunohistochemical analysis demonstrated mTOR was more frequently activated in CCCs than in serous adenocarcinomas (86.6% vs 50%). Treatment with RAD001 markedly inhibited the growth of both RMG1 and KOC7C cells both in vitro and in vivo. Increased expression of phospho-mTOR was observed in cisplatin-resistant RMG1-CR and KOC7C

  1. Efficacy of risk-based, culturally sensitive Ogaa (walleye) consumption advice for Anishinaabe tribal members in the Great Lakes Region.

    PubMed

    DeWeese, Adam D; Kmiecik, Neil E; Chiriboga, Esteban D; Foran, Jeffery A

    2009-05-01

    The Great Lakes Indian Fish and Wildlife Commission (GLIFWC) has produced Ogaa (walleye-Sander vitreus) consumption advisories since 1996 for Anishinaabe from GLIFWC member tribes in the 1837 and 1842 ceded territories of Wisconsin. GLIFWC's advisory maps were revised in 2005 to address cultural sensitivities (to protect tribal lifeways), to utilize recent mercury exposure information, and to incorporate changes in advisory levels for methyl mercury. Lake-specific, risk-based, culturally sensitive consumption advice was provided on color-coded maps for two groups: children under age 15 years and females of childbearing age, and males 15 years and older and females beyond childbearing age. The maps were distributed to, and a behavioral intervention program developed for, the six GLIFWC member tribes in Wisconsin as well as member tribes in Minnesota and the 1842 ceded territory of Michigan. Tribal fish harvesters, tribal health care providers, women of childbearing age or with young children, tribal leaders, elders, and children were targeted specifically for the behavioral intervention. The efficacy of the behavioral intervention was assessed using surveys of 275 tribal fish harvesters from Wisconsin, 139 tribal harvesters from Michigan and Minnesota, and 156 Wisconsin women of childbearing age. Significant increases in the percentage of survey participants who indicated awareness of advisory maps occurred among Wisconsin harvesters (increase from 60% to 77%), Michigan and Minnesota harvesters (29% to 51%), and women of childbearing age in Wisconsin (40% to 87%). A significant increase in preference for smaller Ogaa occurred among tribal harvesters in Wisconsin (41% to 72%) and tribal harvesters in Michigan and Minnesota (49% to 71%), although not among women of childbearing age. The GLIFWC map-based advisory program did not adversely affect tribal harvest of Ogaa, which increased from 63,000 to 88,000 fish in the three states after the intervention.

  2. miRNA-181b increases the sensitivity of pancreatic ductal adenocarcinoma cells to gemcitabine in vitro and in nude mice by targeting BCL-2.

    PubMed

    Cai, Baobao; An, Yong; Lv, Nan; Chen, Jianmin; Tu, Min; Sun, Jie; Wu, Pengfei; Wei, Jishu; Jiang, Kuirong; Miao, Yi

    2013-05-01

    Pancreatic ductal adenocarcinoma (PDAC) is a highly lethal disease and is usually resistant to chemotherapy. MicroRNA‑181b (miR-181b) has been reported to be associated with chemoresistance in various types of cancer. In this study, we investigated the effects of miR-181b on the chemosensitivity of PDAC cells to gemcitabine and the underlying molecular events. miR-181b mimics and inhibitors were synthesized for transient gene transfection in vitro. Lentivirus carrying miR-181b mimics were used to infect PDAC cells for nude mouse xenograft assays by implanting infected PDAC cells into recipient mice. Cell viability was determined by MTT assays, while gene expression was assessed using qRT-PCR, western blot analysis and enzyme-linked immunosorbent assay (ELISA). miR-181b targeting BCL-2 expression was assessed by a dual-luciferase activity assay. The data showed that miRNA-181b expression sensitized PDAC cells to gemcitabine treatment. Although gemcitabine-resistant PDAC cell sublines (SW1990/GR and CFPAC-1/GR) expressed higher levels of miRNA-181b, gemcitabine induced higher levels of apoptosis in PDAC cells transfected with miRNA-181b mimics. The nude mouse xenograft assay data showed that miR-181b transfection also sensitized the cells to gemcitabine treatment in vivo. Molecularly, bioinformatics data predicted that miR-181b was able to bind to BCL-2 mRNA 3'UTR. The dual luciferase activity assay revealed that miRNA-181b downregulated BCL-2 expression. The results from western blot analysis showed a reduced BCL-2 expression following miR-181b transfection but an enhanced caspase-3 activity in miRNA-181b mimic-transfected PDAC cells. This study demonstrates that miRNA-181b sensitizes PDAC cells to gemcitabine by targeting BCL-2.

  3. Hedgehog signaling regulates drug sensitivity by targeting ABC transporters ABCB1 and ABCG2 in epithelial ovarian cancer.

    PubMed

    Chen, Yi; Bieber, Marcia M; Teng, Nelson N H

    2014-08-01

    A major challenge of successful chemotherapy in ovarian cancer is overcoming intrinsic or acquired multi-drug resistance caused by active drug efflux mediated by ATP-binding cassette (ABC) transporters. Regulation of these transporters in ovarian cancer is poorly understood. We have found that abnormal expression of the hedgehog (Hh) signaling pathway transcription factor Gli1 is involved in the regulation of ABC transporters ABCB1 and ABCG2 in ovarian cancer. Hh is a known regulator of cancer cell proliferation and differentiation in several other types of invasive and metastatic malignancies. Our work has demonstrated that Gli1 is abnormally activated in a portion of ovarian cancers. Inhibition of Gli1 expression decreases ABCB1 and ABCG2 gene expression levels and enhances the response of ovarian cancer cells to certain chemotherapeutic drugs. The underlying mechanism is a direct association of Gli1 with a specific consensus sequence located in the promoter region of ABCB1 and ABCG2 genes. This study provides new understanding of ABC gene regulation by Hh signaling pathway, which may lead to the identification of new markers to detect and to anticipate ovarian cancer chemotherapy drug sensitivity.

  4. MiR-23a sensitizes nasopharyngeal carcinoma to irradiation by targeting IL-8/Stat3 pathway

    PubMed Central

    Jia-Quan, Qu; Hong-Mei, Yi; Xu, Ye; Li-Na, Li; Jin-Feng, Zhu; Ta, Xiao; Li, Yuan; Jiao-Yang, Li; Yuan-Yuan, Wang; Juan, Feng; Qiu-Yan, He; Shan-Shan, Lu; Hong, Yi; Zhi-Qiang, Xiao

    2015-01-01

    Radioresistance poses a major challenge in nasopharyngeal carcinoma (NPC) treatment, but little is known about how miRNA regulates this phenomenon. In this study, we investigated the function and mechanism of miR-23a in NPC radioresistance, one of downregulated miRNAs in the radioresistant NPC cells identified by our previous microarray analysis. We observed that miR-23a was frequently downregulated in the radioresistant NPC tissues, and its decrement correlated with NPC radioresistance and poor patient survival, and was an independent predictor for reduced patient survival. In vitro radioresponse assays showed that restoration of miR-23a expression markedly increased NPC cell radiosensitivity. In a mouse model, therapeutic administration of miR-23a agomir dramatically sensitized NPC xenografts to irradiation. Mechanistically, we found that reduced miR-23a promoted NPC cell radioresistance by activating IL-8/Stat3 signaling. Moreover, the levels of IL-8 and phospho-Stat3 were increased in the radioresistance NPC tissues, and negatively associated with miR-23a level. Our data demonstrate that miR-23a is a critical determinant of NPC radioresponse and prognostic predictor for NPC patients, and its decrement enhances NPC radioresistance through activating IL-8/Stat3 signaling, highlighting the therapeutic potential of miR-23a/IL-8/Stat3 signaling axis in NPC radiosensitization. PMID:26314966

  5. An efficient PEGylated liposomal nanocarrier containing cell-penetrating peptide and pH-sensitive hydrazone bond for enhancing tumor-targeted drug delivery

    PubMed Central

    Ding, Yuan; Sun, Dan; Wang, Gui-Ling; Yang, Hong-Ge; Xu, Hai-Feng; Chen, Jian-Hua; Xie, Ying; Wang, Zhi-Qiang

    2015-01-01

    Cell-penetrating peptides (CPPs) as small molecular transporters with abilities of cell penetrating, internalization, and endosomal escape have potential prospect in drug delivery systems. However, a bottleneck hampering their application is the poor specificity for cells. By utilizing the function of hydration shell of polyethylene glycol (PEG) and acid sensitivity of hydrazone bond, we constructed a kind of CPP-modified pH-sensitive PEGylated liposomes (CPPL) to improve the selectivity of these peptides for tumor targeting. In CPPL, CPP was directly attached to liposome surfaces via coupling with stearate (STR) to avoid the hindrance of PEG as a linker on the penetrating efficiency of CPP. A PEG derivative by conjugating PEG with STR via acid-degradable hydrazone bond (PEG2000-Hz-STR, PHS) was synthesized. High-performance liquid chromatography and flow cytometry demonstrated that PHS was stable at normal neutral conditions and PEG could be completely cleaved from liposome surface to expose CPP under acidic environments in tumor. An optimal CPP density on liposomes was screened to guaranty a maximum targeting efficiency on tumor cells as well as not being captured by normal cells that consequently lead to a long circulation in blood. In vitro and in vivo studies indicated, in 4 mol% CPP of lipid modified system, that CPP exerted higher efficiency on internalizing the liposomes into targeted subcellular compartments while remaining inactive and free from opsonins at a maximum extent in systemic circulation. The 4% CPPL as a drug delivery system will have great potential in the clinical application of anticancer drugs in future. PMID:26491292

  6. In situ amplified electrochemical aptasensing for sensitive detection of adenosine triphosphate by coupling target-induced hybridization chain reaction with the assembly of silver nanotags.

    PubMed

    Zhou, Qian; Lin, Youxiu; Lin, Yuping; Wei, Qiaohua; Chen, Guonan; Tang, Dianping

    2016-01-01

    Biomolecular immobilization and construction of the sensing platform are usually crucial for the successful development of a high-efficiency detection system. Herein we report on a novel and label-free signal-amplified aptasensing for sensitive electrochemical detection of small molecules (adenosine triphosphate, ATP, used in this case) by coupling with target-induced hybridization chain reaction (HCR) and the assembly of electroactive silver nanotags. The system mainly consisted of two alternating hairpin probes, a partial-pairing trigger-aptamer duplex DNA and a capture probe immobilized on the electrode. Upon target ATP introduction, the analyte attacked the aptamer and released the trigger DNA, which was captured by capture DNA immobilized on the electrode to form a newly partial-pairing double-stranded DNA. Thereafter, the exposed domain at trigger DNA could be utilized as the initator strand to open the hairpin probes in sequence, and propagated a chain reaction of hybridization events between two alternating hairpins to form a long nicked double-helix. The electrochemical signal derived from the assembled silver nanotags on the nicked double-helix. Under optimal conditions, the electrochemical aptasensor could exhibit a high sensitivity and a low detection limit, and allowed the detection of ATP at a concentration as low as 0.03 pM. Our design showed a high selectivity for target ATP against its analogs because of the high-specificity ATP-aptamer reaction, and its applicable for monitoring ATP in the spiking serum samples. Improtantly, the distinct advantages of the developed aptasensor make it hold a great potential for the development of simple and robust sensing strategies for the detection of other small molecules by controlling the apatmer sequence.

  7. Novel Mad2-targeting miR-493-3p controls mitotic fidelity and cancer cells’ sensitivity to paclitaxel

    PubMed Central

    Mäki-Jouppila, Jenni; Chen, Ping; Elgaaen, Bente Vilming; Straume, Anne Hege; Huhtinen, Kaisa; Cárpen, Olli; Lønning, Per Eystein; Davidson, Ben; Hautaniemi, Sampsa; Kallio, Marko J.

    2016-01-01

    The molecular pathways that contribute to the proliferation and drug response of cancer cells are highly complex and currently insufficiently characterized. We have identified a previously unknown microRNA-based mechanism that provides cancer cells means to stimulate tumorigenesis via increased genomic instability and, at the same time, evade the action of clinically utilized microtubule drugs. We demonstrate miR-493-3p to be a novel negative regulator of mitotic arrest deficient-2 (MAD2), an essential component of the spindle assembly checkpoint that monitors the fidelity of chromosome segregation. The microRNA targets the 3′ UTR of Mad2 mRNA thereby preventing translation of the Mad2 protein. In cancer cells, overexpression of miR-493-3p induced a premature mitotic exit that led to increased frequency of aneuploidy and cellular senescence in the progeny cells. Importantly, excess of the miR-493-3p conferred resistance of cancer cells to microtubule drugs. In human neoplasms, miR-493-3p and Mad2 expression alterations correlated with advanced ovarian cancer forms and high miR-493-3p levels were associated with reduced survival of ovarian and breast cancer patients with aggressive tumors, especially in the paclitaxel therapy arm. Our results suggest that intratumoral profiling of miR-493-3p and Mad2 levels can have diagnostic value in predicting the efficacy of taxane chemotherapy. PMID:26943585

  8. Driver mutations and differential sensitivity to targeted therapies: a new approach to the treatment of lung adenocarcinoma.

    PubMed

    Bronte, Giuseppe; Rizzo, Sergio; La Paglia, Laura; Adamo, Vincenzo; Siragusa, Sergio; Ficorella, Corrado; Santini, Daniele; Bazan, Viviana; Colucci, Giuseppe; Gebbia, Nicola; Russo, Antonio

    2010-11-01

    The adenocarcinoma of the lung has recently shown peculiar molecular characteristics, which relate with both carcinogenesis and response to targeted drugs. Several molecular alterations have been defined as "driver mutations". These are responsible for both the initiation and maintenance of the malignancy. The epidermal growth factor receptor (EGFR) pathway is the main regulator of cell function and cancer development. It has a widely defined role in the occurrence of driver mutations. Up till now EGFR gene mutations, KRAS gene mutations and EML4-ALK fusion genes are the most widely recognized alterations involved in both the biology and the clinical management of lung adenocarcinoma. In this review we report the molecular bases that have led to the clinical application of the detection for such genetic impairments. Subsequently we discuss the clinical studies regarding the prognostic role and the predictive value for response to anti-EGFR tyrosine kinase inhibitors (TKI) of the same mutations. We also provide a potential algorithm as a guide in the choice of the best treatment for patients with adenocarcinoma.

  9. 76 FR 21349 - Science Advisory Board Staff Office; Request for Nominations of Candidates to the EPA's Advisory...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-04-15

    ... AGENCY Science Advisory Board Staff Office; Request for Nominations of Candidates to the EPA's Advisory Council on Clean Air Compliance Analysis (Council) EPA's Clean Air Scientific Advisory Committee (CASAC) and EPA's Science Advisory Board (SAB) AGENCY: Environmental Protection Agency. ACTION:...

  10. Dimeric DNA Aptamer Complexes for High-capacity-targeted Drug Delivery Using pH-sensitive Covalent Linkages.

    PubMed

    Boyacioglu, Olcay; Stuart, Christopher H; Kulik, George; Gmeiner, William H

    2013-07-16

    Treatment with doxorubicin (Dox) results in serious systemic toxicities that limit effectiveness for cancer treatment and cause long-term health issues for cancer patients. We identified a new DNA aptamer to prostate-specific membrane antigen (PSMA) using fixed sequences to promote Dox binding and developed dimeric aptamer complexes (DACs) for specific delivery of Dox to PSMA(+) cancer cells. DACs are stable under physiological conditions and are internalized specifically into PSMA(+) C4-2 cells with minimal uptake into PSMA-null PC3 cells. Cellular internalization of DAC was demonstrated by confocal microscopy and flow cytometry. Covalent modification of DAC with Dox (DAC-D) resulted in a complex with stoichiometry ~4:1. Dox was covalently bound in DAC-D using a reversible linker that promotes covalent attachment of Dox to genomic DNA following cell internalization. Dox was released from the DAC-D under physiological conditions with a half-life of 8 hours, sufficient for in vivo targeting. DAC-D was used to selectively deliver Dox to C4-2 cells with endosomal release and nuclear localization of Dox. DAC-D was selectively cytotoxic to C4-2 cells with similar cytotoxicity as the molar equivalent of free-Dox. In contrast, DAC-D displayed minimal cytotoxicity to PC3 cells, demonstrating the complex displays a high degree of selectivity for PSMA(+) cells. DAC-D displays specificity and stability features that may be useful for improved delivery of Dox selectively to malignant tissue in vivo.Molecular Therapy-Nucleic Acids (2013) 2, e107; doi:10.1038/mtna.2013.37; published online 16 July 2013.

  11. Loss of protein targeting to glycogen sensitizes human hepatocellular carcinoma cells towards glucose deprivation mediated oxidative stress and cell death.

    PubMed

    Yang, Rongqiang; Zhang, Mei; Gustafson, Amber Renee; Wang, Eugenia; Cole, Marsha Paulette; Tooley, Christine Elizabeth Schaner; Cheng, Alan

    2015-05-01

    Protein targeting to glycogen (PTG) is a ubiquitously expressed scaffolding protein that critically regulates glycogen levels in many tissues, including the liver, muscle and brain. However, its importance in transformed cells has yet to be explored in detail. Since recent studies have demonstrated an important role for glycogen metabolism in cancer cells, we decided to assess the effect of PTG levels on the ability of human hepatocellular carcinoma (HepG2) cells to respond to metabolic stress. Although PTG expression did not significantly affect the proliferation of HepG2 cells under normal culture conditions, we determined that PTG plays an important role during glucose deprivation. Overexpression of PTG protected cells from cell death in the absence of glucose, whereas knocking down PTG further promoted cytotoxicity, as measured by the release of lactate dehydrogenase (LDH) into the media. Additionally, we demonstrated that PTG attenuates glucose deprivation induced haeme oxygenase-1 (HO-1) expression, suggesting that PTG protects against glucose deprivation-induced oxidative stress. Indeed, treating cells with the antioxidant N-acetyl cysteine (NAC) rescued cells from cytotoxicity caused by glucose deprivation. Finally, we showed that loss of PTG resulted in enhanced autophagy. In control cells, glucose deprivation suppressed autophagy as determined by the increase in the levels of p62, an autophagy substrate. However, in knockdown cells, this suppression was relieved. Blockade of autophagy also attenuated cytotoxicity from glucose deprivation in PTG knockdown cells. Taken together, our findings identify a novel role for PTG in protecting hepatocellular carcinoma cells from metabolic stress, in part by regulating oxidative stress and autophagy.

  12. Loss of protein targeting to glycogen sensitizes human hepatocellular carcinoma cells towards glucose deprivation mediated oxidative stress and cell death

    PubMed Central

    Yang, Rongqiang; Zhang, Mei; Gustafson, Amber Renee; Wang, Eugenia; Cole, Marsha Paulette; Tooley, Christine Elizabeth Schaner; Cheng, Alan

    2015-01-01

    Protein targeting to glycogen (PTG) is a ubiquitously expressed scaffolding protein that critically regulates glycogen levels in many tissues, including the liver, muscle and brain. However, its importance in transformed cells has yet to be explored in detail. Since recent studies have demonstrated an important role for glycogen metabolism in cancer cells, we decided to assess the effect of PTG levels on the ability of human hepatocellular carcinoma (HepG2) cells to respond to metabolic stress. Although PTG expression did not significantly affect the proliferation of HepG2 cells under normal culture conditions, we determined that PTG plays an important role during glucose deprivation. Overexpression of PTG protected cells from cell death in the absence of glucose, whereas knocking down PTG further promoted cytotoxicity, as measured by the release of lactate dehydrogenase (LDH) into the media. Additionally, we demonstrated that PTG attenuates glucose deprivation induced haeme oxygenase-1 (HO-1) expression, suggesting that PTG protects against glucose deprivation-induced oxidative stress. Indeed, treating cells with the antioxidant N-acetyl cysteine (NAC) rescued cells from cytotoxicity caused by glucose deprivation. Finally, we showed that loss of PTG resulted in enhanced autophagy. In control cells, glucose deprivation suppressed autophagy as determined by the increase in the levels of p62, an autophagy substrate. However, in knockdown cells, this suppression was relieved. Blockade of autophagy also attenuated cytotoxicity from glucose deprivation in PTG knockdown cells. Taken together, our findings identify a novel role for PTG in protecting hepatocellular carcinoma cells from metabolic stress, in part by regulating oxidative stress and autophagy. PMID:26182369

  13. Dimethyl fumarate restores apoptosis sensitivity and inhibits tumor growth and metastasis in CTCL by targeting NF-κB

    PubMed Central

    Müller-Decker, Karin; Schroeder, Anne; Brechmann, Markus; Möbs, Markus; Géraud, Cyrill; Assaf, Chalid; Goerdt, Sergij; Krammer, Peter H.; Gülow, Karsten

    2016-01-01

    Despite intensive efforts in recent years, a curative therapy for cutaneous T-cell lymphoma (CTCL) has not yet been developed. Therefore, the establishment of new therapeutic approaches with higher efficacy rates and milder side effects is strongly desired. A characteristic feature of the malignant T-cell population in CTCL is resistance toward cell death resulting from constitutive NF-κB activation. Therefore, NF-κB–dependent cell death resistance represents an interesting therapeutic target in CTCL because an NF-κB–directed therapy would leave bystander T cells widely unaffected. We investigated the effects of dimethyl fumarate (DMF) on CTCL cells in vitro and in vivo. DMF induced cell death in primary patient-derived CD4+ cells and CTCL cell lines, but hardly in T cells from healthy donors. DMF-induced cell death was linked specifically to NF-κB inhibition. To study the impact of DMF in vivo, we developed 2 CTCL xenograft mouse models with different cutaneous localizations of the T-cell infiltrate. DMF treatment delayed the growth of CTCL tumors and prevented formation of distant metastases. In addition, DMF induced increased cell death in primary CTCL tumors and in liver metastases. In summary, DMF treatment represents a remarkable therapeutic option in CTCL because it restores CTCL apoptosis in vitro and in preclinical models in vivo and prevents spreading of the disease to distant sites. DMF treatment is of particular promise in CTCL because DMF is already in successful clinical use in the treatment of psoriasis and multiple sclerosis allowing fast translation into clinical studies in CTCL. PMID:27268084

  14. Targeting the eIF4E/β-catenin axis sensitizes cervical carcinoma squamous cells to chemotherapy

    PubMed Central

    Xu, Hai; Wang, Zhiyin; Xu, Lang; Mo, Guoyan; Duan, Gangfeng; Wang, Yali; Sun, Zhengang; Chen, Hao

    2017-01-01

    Chemotherapy has improved the clinical outcomes of cervical cancer patients. However, patients develop chemoresistance, whose underlying mechanisms are not well understood. In this study, we investigated the phosphorylation levels of eukaryotic translation initiation factor 4E (eIF4E) in cervical cancer cells subjected to chemotherapy. Results showed that chemotherapeutic drugs significantly increased eIF4E phosphorylation at S209 in HeLa and SiHa cells. Upregulation of phosphorylated eIF4E (p-eIF4E) levels has also been shown in cisplatin-resistant HeLa cells and has been observed to be a common response of cervical cancer patients undergoing chemotherapy. We further showed that chemotherapeutic drugs increase β-catenin activity and mRNA levels of Wnt/β-catenin target genes in cervical cancer cells but not in eIF4E-depleted cells, suggesting that chemotherapeutic drugs activate Wnt/β-catenin signaling in an eIF4E-dependent manner. Inhibiting eIF4E via siRNA knockdown or Wnt/β-catenin using the Wnt inhibitor pyrvinium effectively enhanced the anti-proliferative and pro-apoptotic effects of cisplatin in cervical cancer cells both in vitro and in vivo. Our findings demonstrate that eIF4E/β-catenin signaling plays a positive regulatory role in the resistance of cervical cancer cell to chemotherapy and thus highlight the therapeutic value of eIF4E or β-catenin inhibition in overcoming chemoresistance. PMID:28386346

  15. Development of a rapid, sensitive TaqMan real-time RT-PCR assay for the detection of Rose rosette virus using multiple gene targets.

    PubMed

    Babu, Binoy; Jeyaprakash, Ayyamperumal; Jones, Debra; Schubert, Timothy S; Baker, Carlye; Washburn, Brian K; Miller, Steven H; Poduch, Kristina; Knox, Gary W; Ochoa-Corona, Francisco M; Paret, Mathews L

    2016-09-01

    Rose rosette virus (RRV), belonging to the genus Emaravirus, is a highly destructive pathogen that causes rose rosette disease. The disease is a major concern for the rose industry in the U.S. due to the lack of highly sensitive methods for early detection of RRV. This is critical, as early identification of the infected plants and eradication is necessary in minimizing the risks associated with the spread of the disease. A highly reliable, specific and sensitive detection assay is thus required to test and confirm the presence of RRV in suspected plant samples. In this study a TaqMan real-time reverse transcription-polymerase chain reaction (RT-PCR) assay was developed for the detection of RRV from infected roses, utilizing multiple gene targets. Four pairs of primers and probes; two of them (RRV_2-1 and RRV_2-2) based on the consensus sequences of the glycoprotein gene (RNA2) and the other two (RRV_3-2 and RRV_3-5) based on the nucleocapsid gene (RNA3) were designed. The specificity of the primers and probes was evaluated against other representative viruses infecting roses, belonging to the genera Alfamovirus, Cucumovirus, Ilarvirus, Nepovirus, Tobamovirus, and Tospovirus and one Emaravirus (Wheat mosaic virus). Dilution assays using the in vitro transcripts (spiked with total RNA from healthy plants, and non-spiked) showed that all the primers and probes are highly sensitive in consistently detecting RRV with a detection limit of 1 fg. Testing of the infected plants over a period of time (three times in monthly intervals) indicated high reproducibility, with the primer/probe RRV_3-5 showing 100% positive detection, while RRV_2-1, RRV_2-2 and RRV_3-2 showed 90% positive detection. The developed real-time RT-PCR assay is reliable, highly sensitive, and can be easily used in diagnostic laboratories for testing and confirmation of RRV.

  16. Sensitive detection of Tomato ringspot virus by real-time TaqMan RT-PCR targeting the highly conserved 3'-UTR region.

    PubMed

    Tang, Joe; Khan, Subuhi; Delmiglio, Catia; Ward, Lisa I

    2014-06-01

    A real-time TaqMan RT-PCR assay was developed for the rapid and sensitive detection of Tomato ringspot virus (ToRSV), an important plant virus which infects a wide range of fruit and ornamental crops. Primers and a probe were designed based on the highly conserved 3'-untranslated region (UTR) sequences of ToRSV, to amplify a 182bp fragment of this region of RNA-1 and RNA-2. The assay was demonstrated to reliably amplify all ToRSV isolates tested. The detection limit was estimated to be about 12 copies of the ToRSV target region. No amplification was observed from the RNA of other nepoviruses or healthy host species. A comparison with a published conventional RT-PCR and a SYBR-based qRT-PCR indicated that both of the published assays lacked reliability and sensitivity, as neither were able to amplify all ToRSV isolates tested, and both were approximately 1000 times less sensitive than the novel TaqMan real-time assay. This TaqMan real-time assay was tested using four different reagent kits and was shown to be robust and stable, with no significant differences in sensitivity between kits. It is expected that the implementation of this TaqMan real-time RT-PCR assay will facilitate efficient phytosanitary certification of nursery stock requiring testing for ToRSV by regulatory agencies, and will also have wider uses for the general detection of ToRSV in a range of hosts.

  17. Analysis of the common deletions in the mitochondrial DNA is a sensitive biomarker detecting direct and non-targeted cellular effects of low dose ionizing radiation.

    PubMed

    Schilling-Tóth, Boglárka; Sándor, Nikolett; Kis, Eniko; Kadhim, Munira; Sáfrány, Géza; Hegyesi, Hargita

    2011-11-01

    One of the key issues of current radiation research is the biological effect of low doses. Unfortunately, low dose science is hampered by the unavailability of easily performable, reliable and sensitive quantitative biomarkers suitable detecting low frequency alterations in irradiated cells. We applied a quantitative real time polymerase chain reaction (qRT-PCR) based protocol detecting common deletions (CD) in the mitochondrial genome to assess direct and non-targeted effects of radiation in human fibroblasts. In directly irradiated (IR) cells CD increased with dose and was higher in radiosensitive cells. Investigating conditioned medium-mediated bystander effects we demonstrated that low and high (0.1 and 2Gy) doses induced similar levels of bystander responses and found individual differences in human fibroblasts. The bystander response was not related to the radiosensitivity of the cells. The importance of signal sending donor and signal receiving target cells was investigated by placing conditioned medium from a bystander response positive cell line (F11-hTERT) to bystander negative cells (S1-hTERT) and vice versa. The data indicated that signal sending cells are more important in the medium-mediated bystander effect than recipients. Finally, we followed long term effects in immortalized radiation sensitive (S1-hTERT) and normal (F11-hTERT) fibroblasts up to 63 days after IR. In F11-hTERT cells CD level was increased until 35 days after IR then reduced back to control level by day 49. In S1-hTERT cells the increased CD level was also normalized by day 42, however a second wave of increased CD incidence appeared by day 49 which was maintained up to day 63 after IR. This second CD wave might be the indication of radiation-induced instability in the mitochondrial genome of S1-hTERT cells. The data demonstrated that measuring CD in mtDNA by qRT-PCR is a reliable and sensitive biomarker to estimate radiation-induced direct and non-targeted effects.

  18. 76 FR 14009 - Federal Advisory Committee Act; Technological Advisory Council

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-03-15

    ..., DC 20554. FOR FURTHER INFORMATION CONTACT: Walter Johnston, Chief, Electromagnetic Compatibility Division, 202-418-0807; Walter.Johnston@FCC.gov . SUPPLEMENTARY INFORMATION: Technical Advisory Council... public may submit written comments before the meeting to: Walter Johnston, the FCC's Designated...

  19. 77 FR 70434 - Federal Advisory Committee Act; Technological Advisory Council

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-11-26

    ..., DC 20554. FOR FURTHER INFORMATION CONTACT: Walter Johnston, Chief, Electromagnetic Compatibility Division, 202-418-0807; Walter.Johnston@FCC.gov . SUPPLEMENTARY INFORMATION: The FCC Technological Advisory... written comments before the meeting to: Walter Johnston, the FCC's Designated Federal Officer...

  20. 76 FR 71033 - Federal Advisory Committee Act; Technological Advisory Council

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-11-16

    ..., DC 20554. FOR FURTHER INFORMATION CONTACT: Walter Johnston, Chief, Electromagnetic Compatibility Division, (202) 418-0807; Walter.Johnston@FCC.gov . SUPPLEMENTARY INFORMATION: Technical Advisory Council... written comments before the meeting to: Walter Johnston, the FCC's Designated Federal Officer...

  1. Upregulation of microRNA-524-5p enhances the cisplatin sensitivity of gastric cancer cells by modulating proliferation and metastasis via targeting SOX9

    PubMed Central

    Zhou, Jin; Sun, Qing; Liang, Hansi; Gao, Ling

    2017-01-01

    Cisplatin-based chemotherapy is the most commonly used treatment regimen for gastric cancer (GC), however, the resistance to cisplatin represents the key limitation for the therapeutic efficacy. Aberrant expression of MiR-524-5p appears to be involves in tumorigenesis and chemoresistance. However, the mechanism by which miR-524-5p mediates effects of cisplatin treatment in GC remains poorly understood. Expressions of MiR-524-5p was detected in GC tissues and cell lines by qRT-PCR. Cell proliferation was observed by MTT assay; Cell migration was detected by transwell migration and invasion assay. The targeting protein of miR-524-5p was identified by luciferase reporter assay and western blot. We found that downregulation of miR-524-5p in GC tissues and cell lines. SC-M1 and AZ521 cells resistant to cisplatin expressed low levels of miR-524-5p in comparison to the sensitive parental cells. Overexpression of miR-524-5p expression in SC-M1 and AZ521 cells inhibited cell proliferation, migration, and invasion, and conferred sensitivity to cisplatin-resistant GC cells. Subsequently, we identified SOX9 as a functional target protein of miR-524-5p and found that SOX9 overexpression could counteracts the chemosensitizing effects of miR-524-5p. These results provide novel insight into the regulation of GC tumorigenesis and progression by miRNAs. Restoration of miR-524-5p may have therapeutic potential against GC. PMID:27880941

  2. Retraction Statement: 'MicroRNA-218 increases cellular sensitivity to Rapamycin via targeting Rictor in cervical cancer' by Li J, Wang J, Wang Y, Qiu H.

    PubMed

    2017-02-01

    The above article from APMIS, published online on 24 April 2015 in Wiley Online Library (wileyonlinelibrary.com) and in Volume 123, pp. 562-570, has been retracted by agreement between the authors, the journal Editors in Chief, Professors Bodil Norrild, Ben Vainer and Elisabeth Ralfkiaer, and John Wiley & Sons Ltd. The article has been retracted due to errors in the reported results. In this study, the authors used HeLa and SiHa cell lines to investigate the biological roles of miR-218. However, subsequently it emerged that the two cell lines were contaminated in the laboratory by other unknown cell lines. When repeating the experiments, it was found that the functions of miR-218 were not as significant as had been previously reported, especially its effects on rapamycin sensitivity. Reference Li J, Li X, Wang J, Wang Y, Qiu H. MicroRNA-218 increases cellular sensitivity to Rapamycin via targeting Rictor in cervical cancer. APMIS 2015; 123:562-570. doi: 10.1111/apm.12387.

  3. Bilberry extract (Antho 50) selectively induces redox-sensitive caspase 3-related apoptosis in chronic lymphocytic leukemia cells by targeting the Bcl-2/Bad pathway

    PubMed Central

    Alhosin, Mahmoud; León-González, Antonio J.; Dandache, Israa; Lelay, Agnès; Rashid, Sherzad K.; Kevers, Claire; Pincemail, Joël; Fornecker, Luc-Matthieu; Mauvieux, Laurent; Herbrecht, Raoul; Schini-Kerth, Valérie B.

    2015-01-01

    Defect in apoptosis has been implicated as a major cause of resistance to chemotherapy observed in B cell chronic lymphocytic leukaemia (B CLL). This study evaluated the pro-apoptotic effect of an anthocyanin-rich dietary bilberry extract (Antho 50) on B CLL cells from 30 patients and on peripheral blood mononuclear cells (PBMCs) from healthy subjects, and determined the underlying mechanism. Antho 50 induced concentration- and time-dependent pro-apoptotic effects in B CLL cells but little or no effect in PBMCs. Among the main phenolic compounds of the bilberry extract, delphinidin-3-O-glucoside and delphinidin-3-O-rutinoside induced a pro-apoptotic effect. Antho 50-induced apoptosis is associated with activation of caspase 3, down-regulation of UHRF1, a rapid dephosphorylation of Akt and Bad, and down-regulation of Bcl-2. Antho 50 significantly induced PEG-catalase-sensitive formation of reactive oxygen species in B CLL cells. PEG-catalase prevented the Antho 50-induced induction of apoptosis and related signaling. The present findings indicate that Antho 50 exhibits strong pro-apoptotic activity through redox-sensitive caspase 3 activation-related mechanism in B CLL cells involving dysregulation of the Bad/Bcl-2 pathway. This activity of Antho 50 involves the glucoside and rutinoside derivatives of delphinidin. They further suggest that Antho 50 has chemotherapeutic potential by targeting selectively B CLL cells. PMID:25757575

  4. Targeting the PI3K/AKT pathway via GLI1 inhibition enhanced the drug sensitivity of acute myeloid leukemia cells

    PubMed Central

    Liang, Hui; Zheng, Qi-Li; Fang, Peng; Zhang, Jian; Zhang, Tuo; Liu, Wei; Guo, Min; Robinson, Christopher L.; Chen, Shui-bing; Chen, Xiao-Ping; Chen, Fang-Ping; Zeng, Hui

    2017-01-01

    Combination targeted therapy is commonly used to treat acute myeloid leukemia (AML) patients, particularly in refractory/relapse (RR) population. However, concerns have been raised regarding the safety and patient tolerance of combination chemotherapy. It is critical to choose the appropriate treatment for precision therapy. We performed genome-wide RNA profiling using RNA-Seq to compare the RR group and the complete remission (CR) group (a total of 42 adult AML patients). The Hedgehog (Hh) and PI3K/AKT pathways were upregulated in the RR population, which was further confirmed by western blot and/or qPCR. Overexpression of GLI1 in AML cells led to increased AKT phosphorylation and decreased drug sensitivity, which was attenuated by GLI1 inhibition. By contrast, neither the expression of GLI1 nor apoptosis in response to Ara-C treatment of AML cells was significantly affected by PI3K inhibition. Furthermore, co-inhibition of GLI1 and PI3K induced apoptosis of hematopoietic stem/progenitor cells (HSPCs), which raised serious concerns about the side effects of this treatment. These results indicated that GLI1 inhibition alone, but not combined inhibition, is sufficient to enhance AML drug sensitivity, which provides a novel therapeutic strategy for AML treatment. PMID:28098170

  5. A sensitive quartz crystal microbalance assay of adenosine triphosphate via DNAzyme-activated and aptamer-based target-triggering circular amplification.

    PubMed

    Song, Weiling; Zhu, Zheng; Mao, Yaning; Zhang, Shusheng

    2014-03-15

    In this work, a simple and novel quartz crystal microbalance (QCM) assay is demonstrated to selectively and sensitively detect the adenosine triphosphate (ATP). The amplification process consists of circular nucleic acid strand-displacement polymerization, aptamer recognition strategy and nanoparticle signal amplification. With the involvement of an aptamer-based complex, two amplification reaction templates and AuNP-functionalized probes, the whole circle amplification process is triggered by the target recognition of ATP. As an efficient mass amplifier, AuNP-functionalized probes are introduced to enhance the QCM signals. As a result of DNA multiple amplification, a large number of AuNP-functionalized probes are released and hybridized with the capture probes on the gold electrode. Therefore the QCM signals are significantly enhanced, reaching a detection limit of ATP as low as 1.3 nM. This strategy can be conveniently used for any aptamer-target binding events with other biological detection such as protein and small molecules. Moreover, the practical determination of ATP in cancer cells demonstrates the feasibility of this QCM approach and potential application in clinical diagnostics.

  6. Reaction rate sensitivity of 44Ti production in massive stars and implications of a thick target yield measurement of 40Ca(alpha,gamma)44Ti

    SciTech Connect

    Hoffman, R D; Sheets, S A; Burke, J T; Scielzo, N D; Rauscher, T; Norman, E B; Tumey, S; Brown, T A; Grant, P G; Hurst, A M; Phair, L; Stoyer, M A; Wooddy, T; Fisker, J L; Bleuel, D

    2010-02-16

    We evaluate two dominant nuclear reaction rates and their uncertainties that affect {sup 44}Ti production in explosive nucleosynthesis. Experimentally we develop thick-target yields for the {sup 40}Ca({alpha},{gamma}){sup 44}Ti reaction at E{sub {alpha}} = 4.13, 4.54, and 5.36 MeV using {gamma}-ray spectroscopy. At the highest beam energy, we also performed an activation measurement which agrees with the thick target result. From the measured yields a stellar reaction rate was developed that is smaller than current statistical-model calculations and recent experimental results, which would suggest lower {sup 44}Ti production in scenarios for the {alpha}-rich freeze out. Special attention has been paid to assessing realistic uncertainties of stellar reaction rates produced from a combination of experimental and theoretical cross sections. With such methods, we also develop a re-evaluation of the {sup 44}Ti({alpha},p){sup 47}V reaction rate. Using these two rates we carry out a sensitivity survey of {sup 44}Ti synthesis in eight expansions representing peak temperature and density conditions drawn from a suite of recent supernova explosion models. Our results suggest that the current uncertainty in these two reaction rates could lead to as large an uncertainty in {sup 44}Ti synthesis as that produced by different treatments of stellar physics.

  7. 78 FR 30305 - The President's Management Advisory Board (PMAB); Public Advisory Meeting

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-05-22

    ... work and focus for 2013 which include Management Innovation and Optimizing Federal Real Estate. In... ADMINISTRATION The President's Management Advisory Board (PMAB); Public Advisory Meeting AGENCY: Office of... Management Advisory Board (PMAB), a Federal Advisory Committee established in accordance with the...

  8. 75 FR 33616 - Science Advisory Board Staff Office; Notification of Closed Meetings of the Science Advisory...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-06-14

    ... AGENCY Science Advisory Board Staff Office; Notification of Closed Meetings of the Science Advisory Board... (EPA). ACTION: Notice. SUMMARY: The U.S. Environmental Protection Agency's (EPA), Science Advisory... is: U.S. EPA Science Advisory Board (1400F), U.S. Environmental Protection Agency, 1200...

  9. 75 FR 54871 - Science Advisory Board Staff Office; Notification of a Clean Air Scientific Advisory Committee...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-09-09

    ... AGENCY Science Advisory Board Staff Office; Notification of a Clean Air Scientific Advisory Committee...). ACTION: Notice. SUMMARY: The Environmental Protection Agency (EPA) Science Advisory Board (SAB) Staff... Officer (DFO), EPA Science Advisory Board (1400R), U.S. Environmental Protection Agency, 1200...

  10. Chemical Safety Advisory Committee (CSAC)

    EPA Pesticide Factsheets

    Chemical Safety Advisory Committee (CSAC) provides expert scientific advice, information, and recommendations to the Office of Pollution Prevention and Toxics (OPPT) on the scientific basis for risk assessments, methodologies, and pollution prevention meas

  11. 77 FR 46733 - Marine Fisheries Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-08-06

    ... National Oceanic and Atmospheric Administration RIN 0648-XC145 Marine Fisheries Advisory Committee AGENCY: National Marine Fisheries Service (NMFS), National Oceanic and Atmospheric Administration (NOAA), Commerce... of a forthcoming meeting of the Marine Fisheries Advisory Committee (MAFAC). The members will...

  12. 78 FR 3402 - Marine Fisheries Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-01-16

    ... National Oceanic and Atmospheric Administration RIN 0648-XC443 Marine Fisheries Advisory Committee AGENCY: National Marine Fisheries Service (NMFS), National Oceanic and Atmospheric Administration (NOAA), Commerce... of a forthcoming meeting of the Marine Fisheries Advisory Committee (MAFAC). The members will...

  13. 76 FR 14379 - Marine Fisheries Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-03-16

    ... National Oceanic and Atmospheric Administration RIN 0648-XA265 Marine Fisheries Advisory Committee AGENCY: National Marine Fisheries Service (NMFS), National Oceanic and Atmospheric Administration (NOAA), Commerce... of a forthcoming meeting of the Marine Fisheries Advisory Committee (MAFAC). The members will...

  14. 77 FR 9953 - National Advisory Council

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-02-21

    ... SECURITY Federal Emergency Management Agency National Advisory Council AGENCY: Federal Emergency Management Agency, DHS. ACTION: Committee Management; Request for Applicants for Appointment to the National Advisory Council. SUMMARY: The Federal Emergency Management Agency (FEMA) is requesting individuals who...

  15. 76 FR 22911 - National Advisory Council

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-04-25

    ... SECURITY Federal Emergency Management Agency National Advisory Council AGENCY: Federal Emergency Management Agency, DHS. ACTION: Committee Management; Notice of Federal Advisory Committee Meeting. SUMMARY: The.... Mail: Office of Chief Counsel, Federal Emergency Management Agency (Room 835), 500 C Street,...

  16. 75 FR 67351 - Environmental Management Advisory Board

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-11-02

    ... Environmental Management Advisory Board AGENCY: Department of Energy. ACTION: Notice of Open Teleconference. SUMMARY: This notice announces a teleconference of the Environmental Management Advisory Board (EMAB). The... is to provide the Assistant Secretary for Environmental Management (EM) with advice...

  17. 77 FR 62541 - Investor Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-10-15

    ... From the Federal Register Online via the Government Publishing Office ] SECURITIES AND EXCHANGE COMMISSION Investor Advisory Committee AGENCY: Securities and Exchange Commission. ACTION: Notice of Telephonic Meeting of Securities and Exchange Commission Investor Advisory Committee. SUMMARY: The...

  18. 77 FR 70453 - Chemical Transportation Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-11-26

    ... From the Federal Register Online via the Government Publishing Office DEPARTMENT OF HOMELAND SECURITY Coast Guard Chemical Transportation Advisory Committee AGENCY: Coast Guard, DHS. ] ACTION: Committee Management; Notice of Federal Advisory Committee Meeting. SUMMARY: The Chemical...

  19. 77 FR 10798 - Aviation Rulemaking Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-02-23

    ... TRANSPORTATION Federal Aviation Administration Aviation Rulemaking Advisory Committee AGENCY: Federal Aviation Administration (FAA), DOT. ACTION: Notice of withdrawal of task assignment to the Aviation Rulemaking Advisory Committee (ARAC). SUMMARY: The FAA has withdrawn a task assigned to the Aviation Rulemaking...

  20. 76 FR 39884 - Aviation Security Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-07-07

    ... SECURITY Aviation Security Advisory Committee AGENCY: Transportation Security Administration, DHS. ACTION... Security Administration (TSA) announces the re-establishment of the Aviation Security Advisory Committee...-governmental organizations (NGOs) and stakeholder representatives concerning potential risks to...

  1. 77 FR 57086 - Open Internet Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-09-17

    ... COMMISSION Open Internet Advisory Committee AGENCY: Federal Communications Commission. ACTION: Notice. SUMMARY: The Commission announces the next meeting date, time, and agenda of the Open Internet Advisory... Open Internet rules, and to provide any recommendations it deems appropriate to the...

  2. 78 FR 16852 - Open Internet Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-03-19

    ... COMMISSION Open Internet Advisory Committee AGENCY: Federal Communications Commission. ACTION: Notice. SUMMARY: The Commission announces the next meeting date, time, and agenda of the Open Internet Advisory... Open Internet rules, and to provide any recommendations it deems appropriate to the...

  3. 75 FR 9416 - Advisory Committee Information Hotline

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-03-02

    ... Respiratory Therapy Devices Panel 3014512624 Circulatory System Devices Panel 3014512625 Clinical Chemistry... Central Nervous System Drugs Advisory Committee 3014512543 Pharmaceutical Science & Clinical Pharmacology... current information available about any particular advisory committee meeting, this system will...

  4. 75 FR 13598 - NASA Advisory Council; Meeting

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-03-22

    ... SPACE ADMINISTRATION NASA Advisory Council; Meeting AGENCY: National Aeronautics and Space... Information Technology Infrastructure Committee of the NASA Advisory Council. DATES: Thursday, April 15, 2010...; 2939943. ADDRESSES: NASA Headquarters, 300 E Street, SW., Washington, DC, Room 2O43 FOR...

  5. 77 FR 66476 - National Advisory Council

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-11-05

    ... National Advisory Council will meet by teleconference on Monday, November 19, 2012. The meeting will be open to the public. DATES: The National Advisory Council will meet Monday, November 19, 2012, from...

  6. 76 FR 4089 - Nicolet Resource Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-01-24

    ... From the Federal Register Online via the Government Publishing Office DEPARTMENT OF AGRICULTURE Forest Service Nicolet Resource Advisory Committee AGENCY: Forest Service, USDA. ACTION: Notice of Meeting. SUMMARY: The Nicolet Resource Advisory Committee will meet in Crandon, Wisconsin. The...

  7. 76 FR 770 - Mendocino Resource Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-01-06

    ... From the Federal Register Online via the Government Publishing Office DEPARTMENT OF AGRICULTURE Forest Service Mendocino Resource Advisory Committee AGENCY: Forest Service, USDA. ACTION: Notice of meeting. SUMMARY: The Mendocino County Resource Advisory Committee will meet January 21, 2011 (RAC)...

  8. 77 FR 45329 - Sabine Resource Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-07-31

    ... From the Federal Register Online via the Government Publishing Office DEPARTMENT OF AGRICULTURE Forest Service Sabine Resource Advisory Committee AGENCY: Forest Service, USDA. ACTION: Notice of meeting. SUMMARY: The Sabine Resource Advisory Committee will meet in ] Hemphill, Texas. The committee...

  9. 76 FR 51343 - Mendocino Resource Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-08-18

    ... From the Federal Register Online via the Government Publishing Office DEPARTMENT OF AGRICULTURE Forest Service Mendocino Resource Advisory Committee AGENCY: Forest Service, USDA. ACTION: Notice of meeting. ] SUMMARY: The Mendocino County Resource Advisory Committee will meet September 16, 2011 (RAC)...

  10. 77 FR 47812 - Yakutat Resource Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-08-10

    ... From the Federal Register Online via the Government Publishing Office DEPARTMENT OF AGRICULTURE Forest Service Yakutat Resource Advisory Committee AGENCY: Forest Service, USDA. ACTION: Notice of meeting. SUMMARY: The Yakutat Resource Advisory Committee will meet in Yakutat, Alaska. The committee...

  11. 78 FR 23741 - Flathead Resource Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-04-22

    ... From the Federal Register Online via the Government Publishing Office DEPARTMENT OF AGRICULTURE Forest Service Flathead Resource Advisory Committee AGENCY: Forest Service, USDA. ACTION: Notice of meetings. SUMMARY: The Flathead Resource Advisory Committee will meet in Kalispell, Montana. The...

  12. 77 FR 48126 - Flathead Resource Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-08-13

    ... From the Federal Register Online via the Government Publishing Office DEPARTMENT OF AGRICULTURE Forest Service Flathead Resource Advisory Committee AGENCY: Forest Service, USDA. ACTION: Notice of meetings. SUMMARY: The Flathead Resource Advisory Committee will meet in Kalispell, Montana. The...

  13. 77 FR 49410 - Chequamegon Resource Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-08-16

    ... From the Federal Register Online via the Government Publishing Office DEPARTMENT OF AGRICULTURE Forest Service Chequamegon Resource Advisory Committee AGENCY: Forest Service, USDA. ACTION: Notice of meeting. SUMMARY: The Chequamegon Resource Advisory Committee will meet in Park Falls, Wisconsin....

  14. 77 FR 50676 - Nicolet Resource Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-08-22

    ... From the Federal Register Online via the Government Publishing Office DEPARTMENT OF AGRICULTURE Forest Service Nicolet Resource Advisory Committee AGENCY: Forest Service, USDA. ACTION: Notice of meeting. SUMMARY: The Nicolet Resource Advisory Committee will meet in Crandon, WI. The committee...

  15. 75 FR 52304 - Coconino Resource Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-08-25

    ... Doc No: 2010-21066] DEPARTMENT OF AGRICULTURE Forest Service Coconino Resource Advisory Committee AGENCY: Forest Service, USDA. ACTION: Notice of meeting. SUMMARY: The Coconino Resource Advisory...., Flagstaff, Arizona 86004. Send written comments to Brady Smith. RAC Coordinator, Coconino Resource...

  16. 76 FR 23275 - Allegheny Resource Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-04-26

    ... From the Federal Register Online via the Government Publishing Office DEPARTMENT OF AGRICULTURE Forest Service Allegheny Resource Advisory Committee AGENCY: Forest Service, USDA. ACTION: Notice of meeting. SUMMARY: The Allegheny Resource Advisory Committee will meet in Clarendon, Pennsylvania....

  17. 77 FR 49410 - Kisatchie Resource Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-08-16

    ... From the Federal Register Online via the Government Publishing Office DEPARTMENT OF AGRICULTURE Forest Service Kisatchie Resource Advisory Committee AGENCY: Forest Service, USDA. ACTION: Notice of meeting. SUMMARY: The Kisatchie Resource Advisory Committee will meet in Natchitoches, Louisiana....

  18. 76 FR 20310 - Ketchikan Resource Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-04-12

    ... Forest Service Ketchikan Resource Advisory Committee AGENCY: Forest Service, USDA. ACTION: Notice of meeting. SUMMARY: The Ketchikan Resource Advisory Committee will meet in Ketchikan, Alaska, June 28, 2011... Ranger District, 3031 Tongass Avenue, Ketchikan, Alaska. Send written comments to Ketchikan...

  19. 76 FR 44573 - Chequamegon Resource Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-07-26

    ... From the Federal Register Online via the Government Publishing Office DEPARTMENT OF AGRICULTURE Forest Service Chequamegon Resource Advisory Committee AGENCY: Forest Service, USDA. ACTION: Notice of meeting. SUMMARY: The Chequamegon Resource Advisory Committee will meet in Park Falls, Wisconsin....

  20. 76 FR 22672 - Nicolet Resource Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-04-22

    ... From the Federal Register Online via the Government Publishing Office DEPARTMENT OF AGRICULTURE Forest Service Nicolet Resource Advisory Committee AGENCY: Forest Service, USDA. ACTION: Notice of meeting. SUMMARY: The Nicolet Resource Advisory Committee will meet in Crandon, WI. The committee...

  1. 75 FR 45091 - Ontonagon Resource Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-08-02

    ... From the Federal Register Online via the Government Publishing Office DEPARTMENT OF AGRICULTURE Forest Service Ontonagon Resource Advisory Committee AGENCY: Forest Service, USDA. ACTION: Notice of meeting. SUMMARY: The Ontonagon Resource Advisory Committee will meet in Ontonagon, Michigan....

  2. 78 FR 22842 - Nicolet Resource Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-04-17

    ... From the Federal Register Online via the Government Publishing Office DEPARTMENT OF AGRICULTURE Forest Service Nicolet Resource Advisory Committee AGENCY: Forest Service, USDA. ACTION: Notice of meeting. SUMMARY: The Nicolet Resource Advisory Committee will meet in Crandon, WI. The committee...

  3. 76 FR 50993 - Allegheny Resource Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-08-17

    ... From the Federal Register Online via the Government Publishing Office DEPARTMENT OF AGRICULTURE Forest Service Allegheny Resource Advisory Committee AGENCY: Forest Service, USDA. ACTION: Notice of meeting. SUMMARY: The Allegheny Resource Advisory Committee will meet in Warren, Pennsylvania....

  4. 75 FR 26918 - Fishlake Resource Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-05-13

    ... From the Federal Register Online via the Government Publishing Office DEPARTMENT OF AGRICULTURE Forest Service Fishlake Resource Advisory Committee AGENCY: Forest Service, USDA. ACTION: Notice of meeting. SUMMARY: The Fishlake Resource Advisory Committee will meet in Richfield, Utah. The committee...

  5. 75 FR 3197 - Mendocino Resource Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-01-20

    ... From the Federal Register Online via the Government Publishing Office DEPARTMENT OF AGRICULTURE Forest Service Mendocino Resource Advisory Committee AGENCY: Forest Service, USDA. ACTION: Notice of meeting. SUMMARY: The Mendocino County Resource Advisory Committee will meet February 5, 2010 (RAC)...

  6. 75 FR 55539 - Shoshone Resource Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-09-13

    ... From the Federal Register Online via the Government Publishing Office DEPARTMENT OF AGRICULTURE Forest Service Shoshone Resource Advisory Committee AGENCY: Forest Service, USDA. ACTION: Notice of meeting. SUMMARY: The Shoshone Resource Advisory Committee will meet in Thermopolis, Wyoming....

  7. 76 FR 38109 - Allegheny Resource Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-06-29

    ... From the Federal Register Online via the Government Publishing Office DEPARTMENT OF AGRICULTURE Forest Service Allegheny Resource Advisory Committee AGENCY: Forest Service, USDA. ACTION: Notice of meeting. SUMMARY: The Allegheny Resource Advisory Committee will meet in Marienville, Pennsylvania....

  8. 76 FR 2882 - Ontonagon Resource Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-01-18

    ... From the Federal Register Online via the Government Publishing Office DEPARTMENT OF AGRICULTURE Forest Service Ontonagon Resource Advisory Committee AGENCY: Forest Service, USDA. ACTION: Notice of meeting. SUMMARY: The Ontonagon Resource Advisory Committee will meet in Ewen, Michigan. The Committee...

  9. 76 FR 85 - Coconino Resource Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-01-03

    ... Forest Service Coconino Resource Advisory Committee AGENCY: Forest Service, USDA. ACTION: Notice of meeting. SUMMARY: The Coconino Resource Advisory Committee will meet in Flagstaff, Arizona, to finalize...., Flagstaff, Arizona 86004. Send written comments to Brady Smith, RAC Coordinator, Coconino Resource...

  10. 76 FR 4862 - Fishlake Resource Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-01-27

    ... From the Federal Register Online via the Government Publishing Office DEPARTMENT OF AGRICULTURE Forest Service Fishlake Resource Advisory Committee AGENCY: Forest Service, USDA. ACTION: Notice of meeting. SUMMARY: The Fishlake Resource Advisory Committee will meet in Richfield, Utah. The committee...

  11. 76 FR 22077 - Gogebic Resource Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-04-20

    ... From the Federal Register Online via the Government Publishing Office DEPARTMENT OF AGRICULTURE Forest Service Gogebic Resource Advisory Committee AGENCY: Forest Service, USDA. ACTION: Notice of meeting. SUMMARY: The Gogebic Resource Advisory Committee will meet in Marenisco, Michigan. The...

  12. 77 FR 50460 - Cherokee Resource Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-08-21

    ... From the Federal Register Online via the Government Publishing Office DEPARTMENT OF AGRICULTURE Forest Service Cherokee Resource Advisory Committee AGENCY: Forest Service, USDA. ACTION: Notice of meeting. SUMMARY: The Cherokee Resource Advisory Committee will meet in Knoxville, Tennessee....

  13. 76 FR 45227 - Allegheny Resource Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-07-28

    ... From the Federal Register Online via the Government Publishing Office ] DEPARTMENT OF AGRICULTURE Forest Service Allegheny Resource Advisory Committee AGENCY: Forest Service, USDA. ACTION: Notice of meeting. SUMMARY: The Allegheny Resource Advisory Committee will meet in Clarendon, Pennsylvania....

  14. 76 FR 11193 - Superior Resource Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-03-01

    ... From the Federal Register Online via the Government Publishing Office DEPARTMENT OF AGRICULTURE Forest Service Superior Resource Advisory Committee AGENCY: Forest Service, USDA. ACTION: Notice of meeting. SUMMARY: The Superior Resource Advisory Committee will meet in Duluth, Minnesota. The...

  15. 76 FR 48801 - Gallatin Resource Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-08-09

    ... From the Federal Register Online via the Government Publishing Office DEPARTMENT OF AGRICULTURE Forest Service Gallatin Resource Advisory Committee AGENCY: Forest Service, USDA. ACTION: Notice of meeting. SUMMARY: The Gallatin National Forest's Gallatin Resource Advisory Committee will meet in...

  16. 75 FR 42067 - Chequamegon Resource Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-07-20

    ... From the Federal Register Online via the Government Publishing Office DEPARTMENT OF AGRICULTURE Forest Service Chequamegon Resource Advisory Committee AGENCY: Forest Service, USDA. ACTION: Notice of meeting. SUMMARY: The Nicolet Resource Advisory Committee will meet in Crandon, Wisconsin. The...

  17. 76 FR 56454 - Consumer Advisory Committee Meeting

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-09-13

    ... COMMISSION Consumer Advisory Committee Meeting AGENCY: Federal Communications Commission. ACTION: Notice. SUMMARY: The Commission announces the next meeting date and agenda of its Consumer Advisory Committee (Committee). The purpose of the Committee is to make recommendations to the Commission regarding...

  18. 75 FR 33305 - Consumer Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-06-11

    ... COMMISSION Consumer Advisory Committee AGENCY: Federal Communications Commission. ACTION: Notice. SUMMARY: The Commission announces the next meeting date and agenda of its Consumer Advisory Committee (``Committee''). The purpose of the Committee is to make recommendations to the Commission regarding...

  19. 75 FR 9898 - Consumer Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-03-04

    ... COMMISSION Consumer Advisory Committee AGENCY: Federal Communications Commission. ACTION: Notice. SUMMARY: The Commission announces the next meeting date and agenda of its Consumer Advisory Committee (``Committee''). The purpose of the ] Committee is to make recommendations to the Commission regarding...

  20. 77 FR 55214 - Consumer Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-09-07

    ... COMMISSION Consumer Advisory Committee AGENCY: Federal Communications Commission. ACTION: Notice. SUMMARY: The Commission announces the next meeting date, time, and agenda of its Consumer Advisory Committee... within the jurisdiction of the Commission and to facilitate the participation of all consumers...

  1. 76 FR 3633 - Consumer Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-01-20

    ... COMMISSION Consumer Advisory Committee AGENCY: Federal Communications Commission. ACTION: Notice. SUMMARY: This document announces the rechartering of the Consumer Advisory Committee (hereinafter ``the... ``Commission'') regarding consumer issues within the jurisdiction of the Commission and to facilitate...

  2. 76 FR 45570 - Consumer Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-07-29

    ... COMMISSION Consumer Advisory Committee AGENCY: Federal Communications Commission. ACTION: Notice. SUMMARY: The Commission announces appointment of members and chairperson to its Consumer Advisory Committee... consumers in proceedings before the Commission. DATES: The meeting of the Committee will take place...

  3. 76 FR 65726 - Consumer Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-10-24

    ... COMMISSION Consumer Advisory Committee AGENCY: Federal Communications Commission. ACTION: Notice. SUMMARY: The Commission announces the next meeting date, time, and agenda of its Consumer Advisory Committee... within the jurisdiction of the Commission and to facilitate the participation of all consumers...

  4. 75 FR 41863 - Consumer Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-07-19

    ... COMMISSION Consumer Advisory Committee AGENCY: Federal Communications Commission. ACTION: Notice. SUMMARY: The Commission announces the next meeting date and agenda of its Consumer Advisory Committee (``Committee''). The purpose of the Committee is to make recommendations to the Commission regarding...

  5. 78 FR 54629 - Consumer Advisory Board meeting

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-09-05

    ... From the Federal Register Online via the Government Publishing Office CONSUMER FINANCIAL PROTECTION BUREAU Consumer Advisory Board meeting AGENCY: Bureau of Consumer Financial Protection. ACTION... Consumer Advisory Board (``CAB'' or ``Board'') of the Consumer Financial Protection Bureau (Bureau)....

  6. 75 FR 63830 - Consumer Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-10-18

    ... COMMISSION Consumer Advisory Committee AGENCY: Federal Communications Commission. ACTION: Notice. SUMMARY: The Commission announces the next meeting date and agenda of its Consumer Advisory Committee (``Committee''). The purpose of the Committee is to make recommendations to the Commission regarding...

  7. 75 FR 4819 - Consumer Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-01-29

    ... COMMISSION Consumer Advisory Committee AGENCY: Federal Communications Commission. ACTION: Notice. SUMMARY: The Commission announces the next meeting date and agenda of its Consumer Advisory Committee (``Committee''). The purpose of the Committee is to make recommendations to the Commission regarding...

  8. Practice advisory on liposuction.

    PubMed

    Iverson, Ronald E; Lynch, Dennis J

    2004-04-15

    COMMITTEE STATEMENT: At the 69th annual meeting of the American Society of Plastic Surgeons (ASPS) in October of 2000, the ASPS Board of Directors convened the Task Force on Patient Safety in Office-Based Surgery Facilities. The task force was assembled in the wake of several highly publicized patient deaths involving plastic surgery and increasing state legislative and regulatory activity of office-based surgery facilities. In response to the increased scrutiny of the office-based surgery setting, the task force produced two practice advisories: "Procedures in the Office-Based Surgery Setting" and "Patient Selection in the Office-Based Surgery Setting." Since the task force's inception, professional and public awareness of patient safety issues has continued to grow. This heightened interest resulted in an increased need for plastic surgeons to communicate their views on the topic. To meet this challenge, the task force evolved into the Committee on Patient Safety, allowing the committee to address topics affecting the safety and welfare of plastic surgery patients, regardless of the facility setting. The "Practice Advisory on Liposuction" is the first advisory developed since the committee was formed. It was a lengthy and painstaking process for the committee, which included representatives from related plastic surgery organizations as well as the American Society of Anesthesiologists (ASA). Committee members included Ronald E. Iverson, M.D., chair; Jeffery L. Apfelbaum, M.D., ASA representative; Bruce L. Cunningham, M.D., ASPS/Plastic Surgery Educational Foundation (PSEF) Joint Outcomes Task Force representative; Richard A. D'Amico, M.D., ASPS representative; Victor L. Lewis, Jr., M.D., ASPS Health Policy Analysis Committee representative; Dennis J. Lynch, M.D., ASPS representative; Noel B. McDevitt, M.D., ASPS Deep Vein Thrombosis Task Force representative; Michael F. McGuire, M.D., The American Society for Aesthetic Plastic Surgery (ASAPS) representative

  9. Galactose-installed photo-crosslinked pH-sensitive degradable micelles for active targeting chemotherapy of hepatocellular carcinoma in mice.

    PubMed

    Zou, Yan; Song, Yuan; Yang, Weijing; Meng, Fenghua; Liu, Haiyan; Zhong, Zhiyuan

    2014-11-10

    In this study, we designed and developed galactose-installed photo-crosslinked pH-sensitive degradable micelles (Gal-CLMs) for active targeting chemotherapy of hepatocellular carcinoma in mice. Gal-CLMs were readily obtained from co-self-assembly of poly(ethylene glycol)-b-poly(mono-2,4,6-trimethoxy benzylidene-pentaerythritol carbonate-co-acryloyl carbonate) (PEG-b-P(TMBPEC-co-AC)) and Gal-PEG-b-poly(ε-caprolactone) (Gal-PEG-b-PCL) copolymers followed by photo-crosslinking. Notably, paclitaxel (PTX)-loaded Gal-CLMs (Gal-PTX-CLMs) showed a narrow distribution (PDI=0.08-0.12) with average sizes ranging from 92.1 to 136.3nm depending on the Gal contents. The release of PTX from Gal-CLMs while inhibited at physiological pH was enhanced under endosomal pH conditions. MTT assays in asialoglycoprotein receptor (ASGP-R) over-expressing HepG2 cells demonstrated that half-maximal inhibitory concentration (IC50) values of Gal-PTX-CLMs decreased from 11.7 to 2.9 to 1.1μg/mL with increasing Gal contents from 10% to 20% to 30%, supporting receptor-mediated endocytosis mechanism. The in vivo biodistribution studies in human hepatoma SMMC-7721 tumor-bearing nude mice displayed that Gal20-PTX-CLMs resulted in significantly enhanced drug accumulation in the tumors over non-targeting PTX-CLM counterpart. In accordance, Gal20-PTX-CLMs caused much greater tumor growth inhibition than non-targeting PTX-CLMs as well as non-crosslinking Gal20-PTX-NCLM controls (average tumor volume: ca. 35mm(3)versus 144mm(3) and 130mm(3), respectively). Histological analysis showed that Gal20-PTX-CLMs induced more extensive apoptosis of tumor cells while less damage to normal liver and kidney compared to Taxol. Ligand-installed photo-crosslinked pH-responsive degradable micelles have a great potential for targeted cancer chemotherapy.

  10. Aerospace Safety Advisory Panel

    NASA Astrophysics Data System (ADS)

    1993-03-01

    The Aerospace Safety Advisory Panel (ASAP) provided oversight on the safety aspects of many NASA programs. In addition, ASAP undertook three special studies. At the request of the Administrator, the panel assessed the requirements for an assured crew return vehicle (ACRV) for the space station and reviewed the organization of the safety and mission quality function within NASA. At the behest of Congress, the panel formed an independent, ad hoc working group to examine the safety and reliability of the space shuttle main engine. Section 2 presents findings and recommendations. Section 3 consists of information in support of these findings and recommendations. Appendices A, B, C, and D, respectively, cover the panel membership, the NASA response to the findings and recommendations in the March 1992 report, a chronology of the panel's activities during the reporting period, and the entire ACRV study report.

  11. Aerospace Safety Advisory Panel

    NASA Technical Reports Server (NTRS)

    1993-01-01

    The Aerospace Safety Advisory Panel (ASAP) provided oversight on the safety aspects of many NASA programs. In addition, ASAP undertook three special studies. At the request of the Administrator, the panel assessed the requirements for an assured crew return vehicle (ACRV) for the space station and reviewed the organization of the safety and mission quality function within NASA. At the behest of Congress, the panel formed an independent, ad hoc working group to examine the safety and reliability of the space shuttle main engine. Section 2 presents findings and recommendations. Section 3 consists of information in support of these findings and recommendations. Appendices A, B, C, and D, respectively, cover the panel membership, the NASA response to the findings and recommendations in the March 1992 report, a chronology of the panel's activities during the reporting period, and the entire ACRV study report.

  12. 78 FR 49543 - Towing Safety Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-08-14

    ... SECURITY Coast Guard Towing Safety Advisory Committee AGENCY: Coast Guard, DHS. ACTION: Committee Management; Notice of Federal Advisory Committee Meeting. SUMMARY: The Towing Safety Advisory Committee (TSAC... related to shallow draft inland, coastal waterway navigation and towing safety. The meetings will be...

  13. 78 FR 2950 - Forestry Research Advisory Council

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-01-15

    ... Forest Service Forestry Research Advisory Council AGENCY: Forest Service, USDA. ACTION: Notice of meeting. SUMMARY: The Forestry Research Advisory Council will meet in Washington, DC February 7-8, 2013. The... Apple, Designated Federal Officer, Forestry Research Advisory Council, USDA Forest Service Research...

  14. 75 FR 54854 - Science Advisory Board

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-09-09

    ... National Oceanic and Atmospheric Administration Science Advisory Board AGENCY: Office of Oceanic and... agenda of a forthcoming meeting of the NOAA Science Advisory Board. The members will discuss and provide.... Cynthia Decker, Executive Director, Science Advisory Board, NOAA, Rm. 11230, 1315 East-West...

  15. 77 FR 476 - Science Advisory Board

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-01-05

    ... National Oceanic and Atmospheric Administration (NOAA) Science Advisory Board AGENCY: Office of Oceanic and... agenda of a forthcoming meeting of the NOAA Science Advisory Board. The members will discuss and provide... be reviewed prior to the meeting date. SUPPLEMENTARY INFORMATION: The Science Advisory Board...

  16. 76 FR 2672 - Science Advisory Board Meeting

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-01-14

    ... National Oceanic and Atmospheric Administration Science Advisory Board Meeting AGENCY: Office of Oceanic... agenda of a forthcoming meeting of the NOAA Science Advisory Board. The members will discuss and provide... Decker, Executive Director, Science Advisory Board, NOAA, Rm. 11230, 1315 East-West Highway,...

  17. 77 FR 58356 - Science Advisory Board

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-09-20

    ... National Oceanic and Atmospheric Administration Science Advisory Board AGENCY: Office of Oceanic and... agenda of a forthcoming meeting of the NOAA Science Advisory Board. The members will discuss and provide... the meeting date. SUPPLEMENTARY INFORMATION: The Science Advisory Board (SAB) was established by...

  18. 78 FR 68037 - Science Advisory Board (SAB)

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-11-13

    ... National Oceanic and Atmospheric Administration Science Advisory Board (SAB) AGENCY: Office of Oceanic and... the NOAA Science Advisory Board (SAB) was published in the Federal Register on October 2, 2013.... Cynthia Decker, Executive Director, Science Advisory Board, NOAA, Rm. 11230, 1315 East-West...

  19. 77 FR 4238 - Advisory Committee Management

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-01-27

    ... Part 8 RIN 1400-AC64 Advisory Committee Management AGENCY: Department of State. ACTION: Final rule... establish uniform administrative guidelines and management controls for advisory committees established by... of Subjects in 22 CFR Part 8 Advisory Committee Management. Accordingly, under the authority of 22...

  20. Linchpins or Lost Time: Creating Effective Advisories

    ERIC Educational Resources Information Center

    Johnson, Bil

    2009-01-01

    That advisories in secondary schools are fairly pervasive around the country may be one of the great unintended consequences of the Coalition of Essential Schools (CES) reform effort. While no CES Common Principle explicitly states that advisories should exist in schools, as Coalition Schools evolved, advisories became one of the logical ways for…

  1. 75 FR 53707 - Homeland Security Advisory Council

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-09-01

    ...-53708] [FR Doc No: 2010-21787] DEPARTMENT OF HOMELAND SECURITY [Docket No. DHS-2010-0074] Homeland Security Advisory Council AGENCY: The Office of Policy, DHS. ACTION: Notice of Open Teleconference Federal Advisory Committee Meeting. SUMMARY: The Homeland Security Advisory Council (HSAC) will meet...

  2. 76 FR 4123 - Homeland Security Advisory Council

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-01-24

    ... SECURITY Homeland Security Advisory Council AGENCY: The Office of Policy, DHS. ACTION: Notice of partially closed Federal Advisory Committee Meeting. SUMMARY: The Homeland Security Advisory Council (HSAC) will... . Include docket number in the subject line of the message. Fax: (202) 282-9207 Mail: Homeland...

  3. 78 FR 4120 - Recreation Resource Advisory Committees

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-01-18

    ...; ] DEPARTMENT OF AGRICULTURE Office of the Secretary Recreation Resource Advisory Committees AGENCY: Forest Service, USDA. ACTION: Notice of intent to re-establish the Recreation Resource Advisory Committees... Recreation Resource Advisory Committees (Recreation RACs) pursuant to Section 4 of the Federal...

  4. 78 FR 74174 - Humanities Panel Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-12-10

    ... ARTS AND THE HUMANITIES Humanities Panel Advisory Committee AGENCY: National Endowment for the Humanities. ACTION: Notice of Charter Renewal for Humanities Panel Advisory Committee. SUMMARY: Pursuant to... Humanities Panel advisory committee was renewed for an additional two-year period on November 26, 2013....

  5. 76 FR 2646 - 2010 Census Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-01-14

    ... Bureau of the Census 2010 Census Advisory Committee AGENCY: Bureau of the Census, Department of Commerce. ACTION: Notice of public meeting. SUMMARY: The Bureau of the Census (Census Bureau) is giving notice of a meeting of the 2010 Census Advisory Committee. The Committee will address advisory committee...

  6. 77 FR 26518 - Public Advisory Committees

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-05-04

    ... United States Patent and Trademark Office Public Advisory Committees AGENCY: United States Patent and... three (3)-year terms. The Public Advisory Committee members must be United States citizens and represent..., which, among other things, established two Public Advisory Committees to review the policies,...

  7. 75 FR 15417 - Public Advisory Committees

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-03-29

    ... United States Patent and Trademark Office Public Advisory Committees AGENCY: United States Patent and... three (3)-year terms. The Public Advisory Committee members must be United States citizens and represent..., which, among other things, established two Public Advisory Committees to review the policies,...

  8. 78 FR 55278 - Chemical Transportation Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-09-10

    ... SECURITY Coast Guard Chemical Transportation Advisory Committee AGENCY: Coast Guard, DHS. ACTION: Committee... that the re- establishment of the Chemical Transportation Advisory Committee (CTAC) is necessary and in... Committee: Chemical Transportation Advisory Committee. ADDRESSES: If you desire to submit comments on...

  9. 78 FR 67379 - Chemical Transportation Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-11-12

    ... SECURITY Coast Guard Chemical Transportation Advisory Committee AGENCY: Coast Guard, DHS. ACTION: Committee Management; Notice of Federal Advisory Committee Meeting. SUMMARY: The Chemical Transportation Advisory... hazardous materials c. Update on Bulk Chemical Data Guide (Blue Book) d. Vessel to vessel transfer...

  10. 76 FR 34240 - Chemical Transportation Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-06-13

    ... SECURITY Coast Guard Chemical Transportation Advisory Committee AGENCY: Coast Guard, DHS. ACTION: Committee... that the establishment of the Chemical Transportation Advisory Committee (CTAC) is necessary and in the...: Chemical Transportation Advisory Committee. ADDRESSES: If you desire to submit comments on this...

  11. 76 FR 30955 - Invasive Species Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-05-27

    ... Office of the Secretary Invasive Species Advisory Committee AGENCY: Office of the Secretary, Interior. ACTION: Notice of public meetings of the Invasive Species Advisory Committee. SUMMARY: Pursuant to the provisions of the Federal Advisory Committee Act, notice is hereby given of meetings of the Invasive...

  12. 78 FR 11899 - Invasive Species Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-02-20

    ... Office of the Secretary Invasive Species Advisory Committee AGENCY: Office of the Secretary, Interior... Invasive Species Advisory Committee. The document contained incorrect dates. This document corrects those.... Meeting of the Invasive Species Advisory Committee (OPEN): Thursday, March 7, 2013 through Friday, March...

  13. 76 FR 10577 - Electricity Advisory Committee Meeting

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-02-25

    ... Electricity Advisory Committee Meeting AGENCY: Office of Electricity Delivery and Energy Reliability... reestablished Electricity Advisory Committee (EAC). The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat... of Electricity Delivery and Energy Reliability, U.S. Department of Energy, Forrestal Building,...

  14. 77 FR 10486 - Electricity Advisory Committee Meeting

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-02-22

    ... Electricity Advisory Committee Meeting AGENCY: Office of Electricity Delivery and Energy Reliability... Electricity Advisory Committee (EAC). The Federal Advisory Committee Act (Pub. L. 92- 463, 86 Stat. 770... INFORMATION CONTACT: Matthew Rosenbaum, Office of Electricity Delivery and Energy Reliability, U.S....

  15. 78 FR 9038 - Electricity Advisory Committee Meeting

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-02-07

    ... Electricity Advisory Committee Meeting AGENCY: Office of Electricity Delivery and Energy Reliability... Electricity Advisory Committee (EAC). The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770... FURTHER INFORMATION CONTACT: Matthew Rosenbaum, Office of Electricity Delivery and Energy Reliability,...

  16. 75 FR 61454 - Electricity Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-10-05

    ... Electricity Advisory Committee AGENCY: Department of Energy, Office of Electricity Delivery and Energy...-established DOE Electricity Advisory Committee. The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat...: David Meyer, Designated Federal Officer, Office of Electricity Delivery and Energy Reliability,...

  17. 75 FR 38086 - Technology Advisory Committee Meeting

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-07-01

    ... COMMISSION Technology Advisory Committee Meeting The Commodity Futures Trading Commission's (``Commission'') Technology Advisory Committee will conduct a meeting on Wednesday, July 14, 2010, beginning at 1 p.m. The...://www.cftc.gov . This will be the first meeting of the reestablished Technology Advisory...

  18. 77 FR 15737 - Technology Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-03-16

    ... COMMISSION Technology Advisory Committee AGENCY: Commodity Futures Trading Commission (``CFTC''). ACTION: Notice of meeting of Technology Advisory Committee. SUMMARY: The CFTC announces that on March 29, 2012, the CFTC's Technology Advisory Committee (``TAC'') will hold a public meeting at the CFTC's...

  19. 77 FR 40032 - Methane Hydrate Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-07-06

    ... Methane Hydrate Advisory Committee AGENCY: Office of Fossil Energy, Department of Energy. ACTION: Notice of open meeting. SUMMARY: This notice announces a meeting of the Methane Hydrate Advisory Committee.... SUPPLEMENTARY INFORMATION: Purpose of the Committee: The purpose of the Methane Hydrate Advisory Committee is...

  20. 77 FR 15091 - Environmental Management Advisory Board

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-03-14

    ... Environmental Management Advisory Board AGENCY: Department of Energy. ACTION: Notice of Solicitation of Nominations for Appointment as a member of the Environmental Management Advisory Board. SUMMARY: In accordance... soliciting nominations for candidates to fill vacancies on the Environmental Management Advisory Board...

  1. 76 FR 63664 - Arts Advisory Panel

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-10-13

    ... Doc No: 2011-26421] NATIONAL FOUNDATION ON THE ARTS AND THE HUMANITIES National Endowment for the Arts Arts Advisory Panel Pursuant to Section 10(a)(2) of the Federal Advisory Committee Act (Pub. L. 92-463), as amended, notice is hereby given that thirteen meetings of the Arts Advisory Panel to the...

  2. 76 FR 21772 - Navigation Safety Advisory Council

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-04-18

    ... SECURITY Coast Guard Navigation Safety Advisory Council AGENCY: Coast Guard, DHS. ACTION: Notice of Federal Advisory Committee Meeting. SUMMARY: The Navigation Safety Advisory Council (NAVSAC) will meet on May 4-5...; including the Inland and International Rules of the Road, navigation regulations and equipment,...

  3. 39 CFR 447.31 - Advisory service.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... Advisory Services and Post-Employment Activities § 447.31 Advisory service. (a) The Ethical Conduct Officer is responsible for the administration of the ethics program of the Postal Service. In the exercise of that responsibility, the Ethical Conduct Officer shall coordinate the advisory service provided by...

  4. 77 FR 16894 - Financial Research Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-03-22

    ... Financial Research Advisory Committee AGENCY: Office of Financial Research, Treasury. ACTION: Notice of establishment of the Financial Research Advisory Committee and solicitation of applications for Committee... the Financial Research Advisory Committee. A Charter for the Committee has been prepared and will...

  5. 77 FR 47592 - Ketchikan Resource Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-08-09

    ... Forest Service Ketchikan Resource Advisory Committee AGENCY: Forest Service, USDA. ACTION: Notice of meeting. SUMMARY: The Ketchikan Resource Advisory Committee will meet in Ketchikan, Alaska, September 11..., Ketchikan, Alaska. Send written comments to Ketchikan Resource Advisory Committee, c/o District Ranger,...

  6. 77 FR 54876 - Colville Resource Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-09-06

    ... Forest Service Colville Resource Advisory Committee AGENCY: Forest Service, USDA. ACTION: Notice of meeting. SUMMARY: The Colville Resource Advisory Committee will meet in Colville, Washington. The..., presentation and voting of 2012 Colville Resource Advisory Committee projects. The full agenda may be...

  7. 76 FR 23537 - Resource Advisory Committee Meeting

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-04-27

    ... Resource Advisory Committee Meeting AGENCY: Forest Service, USDA. ACTION: Notice of Public Meeting, Sabine National Forest Resource Advisory Committee. SUMMARY: In accordance with the Secure Rural Schools and..., Sabine National Forest Resource Advisory Committee (RAC) meeting will meet as indicated below. DATES:...

  8. 75 FR 54846 - Sitka Resource Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-09-09

    ... Forest Service Sitka Resource Advisory Committee AGENCY: Forest Service, USDA. ACTION: Notice of meeting. SUMMARY: The Sitka Resource Advisory Committee will meet in Sitka, Alaska, October 8, 2010. The purpose of... Sitka Resource Advisory Committee, c/o District Ranger, USDA Forest Service, 204 Siginaka Way, Sitka,...

  9. 75 FR 54846 - Ketchikan Resource Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-09-09

    ... Forest Service Ketchikan Resource Advisory Committee AGENCY: Forest Service, USDA. ACTION: Notice of meeting. SUMMARY: The Ketchikan Resource Advisory Committee will meet in Ketchikan, Alaska, September 30..., Ketchikan, Alaska. Send written comments to Ketchikan Resource Advisory Committee, c/o District Ranger,...

  10. 75 FR 61693 - Coconino Resource Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-10-06

    ... Forest Service Coconino Resource Advisory Committee AGENCY: Forest Service, USDA. ACTION: Notice of meeting. SUMMARY: The Coconino Resource Advisory Committee will meet in Flagstaff, Arizona. The purpose of the meeting is to receive training on Resource Advisory Committees and National Environmental...

  11. 78 FR 64910 - Virginia Resource Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-10-30

    ... Forest Service Virginia Resource Advisory Committee AGENCY: Forest Service, USDA. ACTION: Notice of meetings. SUMMARY: The Virginia Resource Advisory Committee will meet in Roanoke, Virginia. The committee... 31, 2014: 10 a.m. to 6 p.m. February 7, 2014: 10 a.m. to 6 p.m. All Resource Advisory...

  12. 75 FR 39656 - Coconino Resource Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-07-12

    ... Forest Service Coconino Resource Advisory Committee AGENCY: Forest Service, USDA. ACTION: Notice of meeting. SUMMARY: The Coconino Resource Advisory Committee will meet in Flagstaff, Arizona. The purpose of..., Coconino Resource Advisory Committee, c/o Forest Service, USDA, 1824 S. Thompson St., Flagstaff,...

  13. 76 FR 3081 - Sitka Resource Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-01-19

    ... Forest Service Sitka Resource Advisory Committee AGENCY: Forest Service, USDA. ACTION: Notice of meeting. SUMMARY: The Sitka Resource Advisory Committee will meet in Sitka, Alaska, February 15, 2011. The purpose... to Sitka Resource Advisory Committee, c/o District Ranger, USDA Forest Service, 204 Siginaka...

  14. 75 FR 33241 - Coconino Resource Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-06-11

    ... Forest Service Coconino Resource Advisory Committee AGENCY: Forest Service, USDA. ACTION: Notice of meeting. SUMMARY: The Coconino Resource Advisory Committee will meet in Sedona, Arizona. The purpose of... Smith, RAC Coordinator, Coconino Resource Advisory Committee, c/o Forest Service, USDA, 1824 S....

  15. 75 FR 8916 - Ketchikan Resource Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-02-26

    ... Forest Service Ketchikan Resource Advisory Committee AGENCY: Forest Service, USDA. ACTION: Notice of meeting. SUMMARY: The Ketchikan Resource Advisory Committee will meet in Ketchikan, Alaska, March 31, 2010..., Alaska. Send written comments to Ketchikan Resource Advisory Committee, do District Ranger, USDA...

  16. 75 FR 46902 - Ketchikan Resource Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-08-04

    ... Forest Service Ketchikan Resource Advisory Committee AGENCY: Forest Service, USDA. ACTION: Notice of meeting. SUMMARY: The Ketchikan Resource Advisory Committee will meet in Ketchikan, Alaska, August 18..., Ketchikan, Alaska. Send written comments to Ketchikan Resource Advisory Committee, c/o District Ranger,...

  17. 75 FR 10460 - Yakutat Resource Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-03-08

    ... Forest Service Yakutat Resource Advisory Committee AGENCY: Forest Service, USDA. ACTION: Notice of meeting. SUMMARY: The Yakutat Resource Advisory Committee will meet in Yakutat, Alaska. The purpose of the meeting is to continue business of the Yakutat Resource Advisory Committee. The committee was formed...

  18. 78 FR 45496 - Flathead Resource Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-07-29

    ... Forest Service Flathead Resource Advisory Committee AGENCY: Forest Service, USDA. ACTION: Notice of meetings. SUMMARY: The Flathead Resource Advisory Committee will meet in Kalispell, Montana on the dates..., 2013--4:00 p.m. to 6:30 p.m. All Resource Advisory Committee meetings are subject to change...

  19. MicroRNA-223 Increases the Sensitivity of Triple-Negative Breast Cancer Stem Cells to TRAIL-Induced Apoptosis by Targeting HAX-1

    PubMed Central

    Sun, Xu; Li, Yongqing; Zheng, Meizhu; Zuo, Wenshu; Zheng, Wenzhu

    2016-01-01

    Drug resistance remains a significant challenge in the treatment of triple-negative breast cancer (TNBC). Recent studies have demonstrated that this drug resistance is associated with a group of cells known as cancer stem cells (CSCs), which are believed to determine the sensitivity of tumor cells to cancer treatment. MicroRNAs (miRNAs) are small, non-coding RNAs that play significant roles in normal and cancer cells. MiR-223 reportedly acts as a tumor suppressor in a range of cancers. However, the role of miR-223 in TNBC, especially in triple-negative breast cancer stem cells (TNBCSCs), remains unknown. Here, we found that miR-223 expression was down-regulated in CD44+CD24-/low TNBCSCs compared with non-CSCs. Furthermore, we found that miR-223 overexpression resensitized TNBCSCs to tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-induced apoptosis. The HAX-1 gene, which is located in the mitochondria and functions as an anti-apoptotic protein, was found to be directly regulated by miR-223 in MDA-MB-231 cells. We demonstrated that miR-223 overexpression promoted TRAIL-induced apoptosis through the mitochondria/ROS pathway. In conclusion, our results suggest that miR-223 increases the sensitivity of TNBCSCs to TRAIL-induced apoptosis by targeting HAX-1. Our findings have improved our understanding of the role of miR-223 in TNBC and may contribute to TNBC treatment. PMID:27618431

  20. miR-15a/16 Enhances Radiation Sensitivity of Non-Small Cell Lung Cancer Cells by Targeting the TLR1/NF-κB Signaling Pathway

    SciTech Connect

    Lan, Fengming; Yue, Xiao; Ren, Gang; Li, Hongqi; Ping, Li; Wang, Yingjie; Xia, Tingyi

    2015-01-01

    Purpose: Many miRNAs have been identified as essential issues and core determining factors in tumor radiation. Recent reports have demonstrated that miRNAs and Toll-like receptors could exert reciprocal effects to control cancer development in various ways. However, a novel role of miR-15a/16 in enhancing radiation sensitivity by directly targeting TLR1 has not been reported, to our knowledge. Methods and Materials: Bioinformatic analyses, luciferase reporter assay, biochemical assays, and subcutaneous tumor establishment were used to characterize the signaling pathways of miRNA-15a/16 in response to radiation treatment. Results: First, an inverse correlation between the expression of miR-15a/16 and TLR1 protein was revealed in non-small cell lung cancer (NSCLC) and normal lung tissues. Next, we corroborated that miR-15a/16 specifically bound to TLR1 3′UTR and inhibited the expression of TLR1 in H358 and A549 cells. Furthermore, miR-15a/16 downregulated the activity of the NF-κB signaling pathway through TLR1. In addition, overexpression of miR-15a/16 inhibited survival capability and increased radiation-induced apoptosis, resulting in enhancement of radiosensitivity in H358 and A549 cells. Finally, subcutaneous tumor bearing NSCLC cells in a nude mice model was established, and the results showed that combined groups (miR-15a/16 + radiation) inhibited tumor growth more significantly than did radiation alone. Conclusions: We mainly elucidate that miRNA-15a/16 can enhance radiation sensitivity by regulating the TLR1/NF-κB signaling pathway and act as a potential therapeutic approach to overcome radioresistance for lung cancer treatment.

  1. Targeting ACLY sensitizes castration-resistant prostate cancer cells to AR antagonism by impinging on an ACLY-AMPK-AR feedback mechanism.

    PubMed

    Shah, Supriya; Carriveau, Whitney J; Li, Jinyang; Campbell, Sydney L; Kopinski, Piotr K; Lim, Hee-Woong; Daurio, Natalie; Trefely, Sophie; Won, Kyoung-Jae; Wallace, Douglas C; Koumenis, Constantinos; Mancuso, Anthony; Wellen, Kathryn E

    2016-07-12

    The androgen receptor (AR) plays a central role in prostate tumor growth. Inappropriate reactivation of the AR after androgen deprivation therapy promotes development of incurable castration-resistant prostate cancer (CRPC). In this study, we provide evidence that metabolic features of prostate cancer cells can be exploited to sensitize CRPC cells to AR antagonism. We identify a feedback loop between ATP-citrate lyase (ACLY)-dependent fatty acid synthesis, AMPK, and the AR in prostate cancer cells that could contribute to therapeutic resistance by maintaining AR levels. When combined with an AR antagonist, ACLY inhibition in CRPC cells promotes energetic stress and AMPK activation, resulting in further suppression of AR levels and target gene expression, inhibition of proliferation, and apoptosis. Supplying exogenous fatty acids can restore energetic homeostasis; however, this rescue does not occur through increased β-oxidation to support mitochondrial ATP production. Instead, concurrent inhibition of ACLY and AR may drive excess ATP consumption as cells attempt to cope with endoplasmic reticulum (ER) stress, which is prevented by fatty acid supplementation. Thus, fatty acid metabolism plays a key role in coordinating ER and energetic homeostasis in CRPC cells, thereby sustaining AR action and promoting proliferation. Consistent with a role for fatty acid metabolism in sustaining AR levels in prostate cancer in vivo, AR mRNA levels in human prostate tumors correlate positively with expression of ACLY and other fatty acid synthesis genes. The ACLY-AMPK-AR network can be exploited to sensitize CRPC cells to AR antagonism, suggesting novel therapeutic opportunities for prostate cancer.

  2. SYBR Green real-time PCR-RFLP assay targeting the plasmodium cytochrome B gene--a highly sensitive molecular tool for malaria parasite detection and species determination.

    PubMed

    Xu, Weiping; Morris, Ulrika; Aydin-Schmidt, Berit; Msellem, Mwinyi I; Shakely, Delér; Petzold, Max; Björkman, Anders; Mårtensson, Andreas

    2015-01-01

    A prerequisite for reliable detection of low-density Plasmodium infections in malaria pre-elimination settings is the availability of ultra-sensitive and high-throughput molecular tools. We developed a SYBR Green real-time PCR restriction fragment length polymorphism assay (cytb-qPCR) targeting the cytochrome b gene of the four major human Plasmodium species (P. falciparum, P. vivax, P. malariae, and P. ovale) for parasite detection and species determination with DNA extracted from dried blood spots collected on filter paper. The performance of cytb-qPCR was first compared against four reference PCR methods using serially diluted Plasmodium samples. The detection limit of the cytb-qPCR was 1 parasite/μl (p/μl) for P. falciparum and P. ovale, and 2 p/μl for P. vivax and P. malariae, while the reference PCRs had detection limits of 0.5-10 p/μl. The ability of the PCR methods to detect low-density Plasmodium infections was then assessed using 2977 filter paper samples collected during a cross-sectional survey in Zanzibar, a malaria pre-elimination setting in sub-Saharan Africa. Field samples were defined as 'final positive' if positive in at least two of the five PCR methods. Cytb-qPCR preformed equal to or better than the reference PCRs with a sensitivity of 100% (65/65; 95%CI 94.5-100%) and a specificity of 99.9% (2910/2912; 95%CI 99.7-100%) when compared against 'final positive' samples. The results indicate that the cytb-qPCR may represent an opportunity for improved molecular surveillance of low-density Plasmodium infections in malaria pre-elimination settings.

  3. RBM3 regulates temperature sensitive miR-142-5p and miR-143 (thermomiRs), which target immune genes and control fever.

    PubMed

    Wong, Justin J-L; Au, Amy Y M; Gao, Dadi; Pinello, Natalia; Kwok, Chau-To; Thoeng, Annora; Lau, Katherine A; Gordon, Jane E A; Schmitz, Ulf; Feng, Yue; Nguyen, Trung V; Middleton, Robert; Bailey, Charles G; Holst, Jeff; Rasko, John E J; Ritchie, William

    2016-04-07

    Fever is commonly used to diagnose disease and is consistently associated with increased mortality in critically ill patients. However, the molecular controls of elevated body temperature are poorly understood. We discovered that the expression of RNA-binding motif protein 3 (RBM3), known to respond to cold stress and to modulate microRNA (miRNA) expression, was reduced in 30 patients with fever, and in THP-1-derived macrophages maintained at a fever-like temperature (40 °C). Notably, RBM3 expression is reduced during fever whether or not infection is demonstrable. Reduced RBM3 expression resulted in increased expression of RBM3-targeted temperature-sensitive miRNAs, we termed thermomiRs. ThermomiRs such as miR-142-5p and miR-143 in turn target endogenous pyrogens including IL-6, IL6ST, TLR2, PGE2 and TNF to complete a negative feedback mechanism, which may be crucial to prevent pathological hyperthermia. Using normal PBMCs that were exogenously exposed to fever-like temperature (40 °C), we further demonstrate the trend by which decreased levels of RBM3 were associated with increased levels of miR-142-5p and miR-143 and vice versa over a 24 h time course. Collectively, our results indicate the existence of a negative feedback loop that regulates fever via reduced RBM3 levels and increased expression of miR-142-5p and miR-143.

  4. Sequence and overexpression of GPP130/GIMPc: evidence for saturable pH-sensitive targeting of a type II early Golgi membrane protein.

    PubMed Central

    Linstedt, A D; Mehta, A; Suhan, J; Reggio, H; Hauri, H P

    1997-01-01

    It is thought that residents of the Golgi stack are localized by a retention mechanism that prevents their forward progress. Nevertheless, some early Golgi proteins acquire late Golgi modifications. Herein, we describe GPP130 (Golgi phosphoprotein of 130 kDa), a 130-kDa phosphorylated and glycosylated integral membrane protein localized to the cis/medial Golgi. GPP130 appears to be the human counterpart of rat Golgi integral membrane protein, cis (GIMPc), a previously identified early Golgi antigen that acquires late Golgi carbohydrate modifications. The sequence of cDNAs encoding GPP130 indicate that it is a type II membrane protein with a predicted molecular weight of 81,880 and an unusually acidic lumenal domain. On the basis of the alignment with several rod-shaped proteins and the presence of multiple predicted coiled-coil regions, GPP130 may form a flexible rod in the Golgi lumen. In contrast to the behavior of previously studied type II Golgi proteins, overexpression of GPP130 led to a pronounced accumulation in endocytotic vesicles, and endogenous GPP130 reversibly redistributed to endocytotic vesicles after chloroquine treatment. Thus, localization of GPP130 to the early Golgi involves steps that are saturable and sensitive to lumenal pH, and GPP130 contains targeting information that specifies its return to the Golgi after chloroquine washout. Given that GIMPc acquires late Golgi modifications in untreated cells, it seems likely that GPP130/GIMPc continuously cycles between the early Golgi and distal compartments and that an unidentified retrieval mechanism is important for its targeting. Images PMID:9201717

  5. RBM3 regulates temperature sensitive miR-142–5p and miR-143 (thermomiRs), which target immune genes and control fever

    PubMed Central

    Wong, Justin J.-L.; Au, Amy Y.M.; Gao, Dadi; Pinello, Natalia; Kwok, Chau-To; Thoeng, Annora; Lau, Katherine A.; Gordon, Jane E.A.; Schmitz, Ulf; Feng, Yue; Nguyen, Trung V.; Middleton, Robert; Bailey, Charles G.; Holst, Jeff; Rasko, John E.J.; Ritchie, William

    2016-01-01

    Fever is commonly used to diagnose disease and is consistently associated with increased mortality in critically ill patients. However, the molecular controls of elevated body temperature are poorly understood. We discovered that the expression of RNA-binding motif protein 3 (RBM3), known to respond to cold stress and to modulate microRNA (miRNA) expression, was reduced in 30 patients with fever, and in THP-1-derived macrophages maintained at a fever-like temperature (40°C). Notably, RBM3 expression is reduced during fever whether or not infection is demonstrable. Reduced RBM3 expression resulted in increased expression of RBM3-targeted temperature-sensitive miRNAs, we termed thermomiRs. ThermomiRs such as miR-142–5p and miR-143 in turn target endogenous pyrogens including IL-6, IL6ST, TLR2, PGE2 and TNF to complete a negative feedback mechanism, which may be crucial to prevent pathological hyperthermia. Using normal PBMCs that were exogenously exposed to fever-like temperature (40°C), we further demonstrate the trend by which decreased levels of RBM3 were associated with increased levels of miR-142–5p and miR-143 and vice versa over a 24 h time course. Collectively, our results indicate the existence of a negative feedback loop that regulates fever via reduced RBM3 levels and increased expression of miR-142–5p and miR-143. PMID:26825461

  6. The effect of dietary fish oil on weight gain and insulin sensitivity is dependent on APOE genotype in humanized targeted replacement mice

    PubMed Central

    Slim, Kenna E.; Vauzour, David; Tejera, Noemi; Voshol, Peter J.; Cassidy, Aedin; Minihane, Anne Marie

    2017-01-01

    We investigated the independent and interactive impact of the common APOE genotype and marine n-3 polyunsaturated fatty acids (PUFAs) on the development of obesity and associated cardiometabolic dysfunction in a murine model. Human APOE3 and APOE4 targeted replacement mice were fed either a control high-fat diet (HFD) or an HFD supplemented with 3% n-3 PUFAs from fish oil (HFD + FO) for 8 wk. We established the impact of intervention on food intake, body weight, and visceral adipose tissue (VAT) mass; plasma, lipids (cholesterol and triglycerides), liver enzymes, and adipokines; glucose and insulin during an intraperitoneal glucose tolerance test; and Glut4 and ApoE expression in VAT. HFD feeding induced more weight gain and higher plasma lipids in APOE3 compared to APOE4 mice (P < 0.05), along with a 2-fold higher insulin and impaired glucose tolerance. Supplementing APOE3, but not APOE4, animals with dietary n-3 PUFAs decreased body-weight gain, plasma lipids, and insulin (P < 0.05) and improved glucose tolerance, which was associated with increased VAT Glut4 mRNA levels (P < 0.05). Our findings demonstrate that an APOE3 genotype predisposes mice to develop obesity and its metabolic complications, which was attenuated by n-3 PUFA supplementation.—Slim, K. E., Vauzour, D., Tejera, N., Voshol, P. J., Cassidy, A., Minihane, A. M. The effect of dietary fish oil on weight gain and insulin sensitivity is dependent on APOE genotype in humanized targeted replacement mice. PMID:27895108

  7. An Analog-sensitive Version of the Protein Kinase Slt2 Allows Identification of Novel Targets of the Yeast Cell Wall Integrity Pathway*

    PubMed Central

    Alonso-Rodríguez, Esmeralda; Fernández-Piñar, Pablo; Sacristán-Reviriego, Almudena; Molina, María; Martín, Humberto

    2016-01-01

    The yeast cell wall integrity MAPK Slt2 mediates the transcriptional response to cell wall alterations through phosphorylation of transcription factors Rlm1 and SBF. However, the variety of cellular functions regulated by Slt2 suggests the existence of a significant number of still unknown substrates for this kinase. To identify novel Slt2 targets, we generated and characterized an analog-sensitive mutant of Slt2 (Slt2-as) that can be specifically inhibited by bulky kinase inhibitor analogs. We demonstrated that Slt2-as is able to use adenosine 5′-[γ-thio]triphosphate analogs to thiophosphorylate its substrates in yeast cell extracts as well as when produced as recombinant proteins in Escherichia coli. Taking advantage of this chemical-genetic approach, we found that Slt2 phosphorylates the MAPK phosphatase Msg5 both in the N-terminal regulatory and C-terminal catalytic domains. Moreover, we identified the calcineurin regulator Rcn2, the 4E-BP (translation initiation factor eIF4E-binding protein) translation repressor protein Caf20, and the Golgi-associated adaptor Gga1 as novel targets for Slt2. The Slt2 phosphorylation sites on Rcn2 and Caf20 were determined. We also demonstrated that, in the absence of SLT2, the GGA1 paralog GGA2 is essential for cells to survive under cell wall stress and for proper protein sorting through the carboxypeptidase Y pathway. Therefore, Slt2-as provides a powerful tool that can expand our knowledge of the outputs of the cell wall integrity MAPK pathway. PMID:26786099

  8. STAT3 is essential for Hodgkin lymphoma cell proliferation and is a target of tyrphostin AG17 which confers sensitization for apoptosis.

    PubMed

    Holtick, U; Vockerodt, M; Pinkert, D; Schoof, N; Stürzenhofecker, B; Kussebi, N; Lauber, K; Wesselborg, S; Löffler, D; Horn, F; Trümper, L; Kube, D

    2005-06-01

    Classical Hodgkin lymphoma (cHL) is a distinct malignancy of the immune system. Despite the progress made in the understanding of the biology of cHL, the transforming events remain to be elucidated. Recently, we demonstrated that the Janus kinase inhibitor AG490 blocked cellular proliferation and STAT3 phosphorylation in cHL. To explore the potential of constitutively activated STAT3 as a drug target and its role in cHL pathogenesis, different cHL cell lines were analyzed. Treatment of cHL cells by the protein tyrosine kinase inhibitor AG17 was associated with inhibition of cellular proliferation and cell cycle arrest. AG17 treatment was accompanied by decreased levels of STAT3 phosphorylation, whereas NF-kappaB and p38/SAPK2 signaling were not inhibited. Incubation with AG17 or AG490 sensitized cHL cells to CD95/Fas/Apo-1 or staurosporine-mediated apoptosis. Coincubation of tyrphostins with staurosporine was accompanied by rapid complete inhibition of STAT3 phosphorylation. RNA interference directed against STAT3 in L428 and L1236 cHL cells demonstrated that STAT3 is essential for cell proliferation of these cHL cells. In conclusion, these findings support the concept that STAT3 signaling is important in the pathogenesis of cHL and tyrphostins are agents for developing new therapeutic strategies.

  9. Development of a Rapid, Sensitive, and Field-Deployable Razor Ex BioDetection System and Quantitative PCR Assay for Detection of Phymatotrichopsis omnivora Using Multiple Gene Targets

    PubMed Central

    Arif, M.; Marek, S. M.; Melcher, U.

    2013-01-01

    A validated, multigene-based method using real-time quantitative PCR (qPCR) and the Razor Ex BioDetection system was developed for detection of Phymatotrichopsis omnivora. This soilborne fungus causes Phymatotrichopsis root rot of cotton, alfalfa, and other dicot crops in the southwestern United States and northern Mexico, leading to significant crop losses and limiting the range of crops that can be grown in soils where the fungus is established. It is on multiple lists of regulated organisms. Because P. omnivora is difficult to isolate, accurate and sensitive culture-independent diagnostic tools are needed to confirm infections by this fungus. Specific PCR primers and probes were designed based on P. omnivora nucleotide sequences of the genes encoding rRNA internal transcribed spacers, beta-tubulin, and the second-largest subunit of RNA polymerase II (RPB2). PCR products were cloned and sequenced to confirm their identity. All primer sets allowed early detection of P. omnivora in infected but asymptomatic plants. A modified rapid DNA purification method, which facilitates a quick (∼30-min) on-site assay capability for P. omnivora detection, was developed. Combined use of three target genes increased the assay accuracy and broadened the range of detection. To our knowledge, this is the first report of a multigene-based, field-deployable, rapid, and reliable identification method for a fungal plant pathogen and should serve as a model for the development of field-deployable assays of other phytopathogens. PMID:23354717

  10. The Core Promoter and Redox-sensitive Cis-elements as Key Targets for Inactivation of the Lysyl Oxidase Gene by Cadmium

    PubMed Central

    Li, Jianmin; Cheng, Guang; Zheng, Maoguen; Zhao, Yinzhi; Zhou, Jing; Li, Wande

    2015-01-01

    Exposure of humans to cadmium (Cd) either from environmental contamination or from cigarette smoke, often induces lung emphysema and cancers. Lysyl oxidase (LOX), a copper-dependent enzyme essential for crosslinking of the extracellular matrix, displays antagonistic effects on emphysema and cancer pathogenesis. Our previous studies showed down-regulation of LOX in Cd-resistant (CdR) rat fetal lung fibroblasts (RFL6) derived from parental cells via long-term Cd exposure. The cloned rat LOX gene promoter −804/−1 (relative to ATG) with the maximal promoter activity contains the Inr-DPE core promoter, putative NFI binding sites, metal response elements (MRE) and antioxidant response elements (ARE). ChIP assays reported here further characterize the rat LOX gene promoter in response to Cd. CdR cells exhibited enhanced methylation of CpG at the LOX core promoter region and reduced activities of the NFI binding sites and MRE, but increased activity of the ARE in a dose-dependent manner. The collective effect of Cd on the LOX promoter is trans-inhibition of the LOX gene as shown by suppression of histone H3 acetylation in the LOX core promoter region. Thus, the LOX core promoter and redox-sensitive cis-elements are key Cd targets for down-regulation of LOX relevant to mechanisms for Cd-induced emphysema and lung cancers. PMID:25741534

  11. 21 CFR 14.90 - Ad hoc advisory committee members.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 1 2011-04-01 2011-04-01 false Ad hoc advisory committee members. 14.90 Section... PUBLIC HEARING BEFORE A PUBLIC ADVISORY COMMITTEE Members of Advisory Committees § 14.90 Ad hoc advisory committee members. In selecting members of an ad hoc advisory committee, the Commissioner may use...

  12. 21 CFR 14.90 - Ad hoc advisory committee members.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 1 2010-04-01 2010-04-01 false Ad hoc advisory committee members. 14.90 Section... PUBLIC HEARING BEFORE A PUBLIC ADVISORY COMMITTEE Members of Advisory Committees § 14.90 Ad hoc advisory committee members. In selecting members of an ad hoc advisory committee, the Commissioner may use...

  13. 22 CFR 214.32 - Calling of advisory committee meetings.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 22 Foreign Relations 1 2010-04-01 2010-04-01 false Calling of advisory committee meetings. 214.32 Section 214.32 Foreign Relations AGENCY FOR INTERNATIONAL DEVELOPMENT ADVISORY COMMITTEE MANAGEMENT Operation of Advisory Committees § 214.32 Calling of advisory committee meetings. (a) No advisory...

  14. Multiple nickel-sensitive targets elicit cardiac arrhythmia in isolated mouse hearts after pituitary adenylate cyclase-activating polypeptide-mediated chronotropy.

    PubMed

    Tevoufouet, Etienne E; Nembo, Erastus N; Distler, Fabian; Neumaier, Felix; Hescheler, Jürgen; Nguemo, Filomain; Schneider, Toni

    2017-03-01

    The pituitary adenylate cyclase-activating polypeptide (PACAP)-27 modulates various biological processes, from the cellular level to function specification. However, the cardiac actions of this neuropeptide are still under intense studies. Using control (+|+) and mice lacking (-|-) either R-type (Cav2.3) or T-type (Cav3.2) Ca(2+) channels, we investigated the effects of PACAP-27 on cardiac activity of spontaneously beating isolated perfused hearts. Superfusion of PACAP-27 (20nM) caused a significant increase of baseline heart frequency in Cav2.3(+|+) (156.9±10.8 to 239.4±23.4 bpm; p<0.01) and Cav2.3(-|-) (190.3±26.4 to 270.5±25.8 bpm; p<0.05) hearts. For Cav3.2, the heart rate was significantly increased in Cav3.2(-|-) (133.1±8.5 bpm to 204.6±27.9 bpm; p<0.05) compared to Cav3.2(+|+) hearts (185.7±11.2 bpm to 209.3±22.7 bpm). While the P wave duration and QTc interval were significantly increased in Cav2.3(+|+) and Cav2.3(-|-) hearts following PACAP-27 superfusion, there was no effect in Cav3.2(+|+) and Cav3.2(-|-) hearts. The positive chronotropic effects observed in the four study groups, as well as the effect on P wave duration and QTc interval were abolished in the presence of Ni(2+) (50μM) and PACAP-27 (20nM) in hearts from Cav2.3(+|+) and Cav2.3(-|-) mice. In addition to suppressing PACAP's response, Ni(2+) also induced conduction disturbances in investigated hearts. In conclusion, the most Ni(2+)-sensitive Ca(2+) channels (R- and T-type) may modulate the PACAP signaling cascade during cardiac excitation in isolated mouse hearts, albeit to a lesser extent than other Ni(2+)-sensitive targets.

  15. Sensitivity of fetal RHD screening for safe guidance of targeted anti-D immunoglobulin prophylaxis: prospective cohort study of a nationwide programme in the Netherlands

    PubMed Central

    de Haas, Masja; Thurik, Florentine F; van der Ploeg, Catharina P B; Veldhuisen, Barbera; Hirschberg, Hoang; Soussan, Aicha Ait; Woortmeijer, Heleen; Abbink, Frithjofna; Page-Christiaens, Godelieve C M L; Scheffer, Peter G; Ellen van der Schoot, C

    2016-01-01

    Objective To determine the accuracy of non-invasive fetal testing for the RHD gene in week 27 of pregnancy as part of an antenatal screening programme to restrict anti-D immunoglobulin use to women carrying a child positive for RHD. Design Prospectively monitoring of fetal RHD testing accuracy compared with serological cord blood typing on introduction of the test. Fetal RHD testing was performed with a duplex real time quantitative polymerase chain reaction, with cell-free fetal DNA isolated from 1 mL of maternal plasma The study period was between 4 July 2011 and 7 October 2012. The proportion of women participating in screening was determined. Setting Nationwide screening programme, the Netherlands. Tests are performed in a centralised setting. Participants 25 789 RhD negative pregnant women. Main outcome measures Sensitivity, specificity, false negative rate, and false positive rate of fetal RHD testing compared with serological cord blood typing; proportion of technical failures; and compliance to the screening programme. Results A fetal RHD test result and serological cord blood result were available for 25 789 pregnancies. Sensitivity for detection of fetal RHD was 99.94% (95% confidence interval 99.89% to 99.97%) and specificity was 97.74% (97.43% to 98.02%). Nine false negative results for fetal RHD testing were registered (0.03%, 95% confidence interval 0.01% to 0.06%). In two cases these were due to technical failures. False positive fetal RHD testing results were registered for 225 samples (0.87%, 0.76% to 0.99%). Weak RhD expression was shown in 22 of these cases, justifying anti-D immunoglobulin use. The negative and positive predictive values were 99.91% (95% confidence interval 99.82% to 99.95%) and 98.60% (98.40% to 98.77%), respectively. More than 98% of the women participated in the screening programme. Conclusions Fetal RHD testing in week 27 of pregnancy as part of a national antenatal screening programme is highly reliable and can

  16. MicroRNA-223 Enhances Radiation Sensitivity of U87MG Cells In Vitro and In Vivo by Targeting Ataxia Telangiectasia Mutated

    SciTech Connect

    Liang, Liping; Zhu, Ji; Zaorsky, Nicholas G.; Deng, Yun; Wu, Xingzhong; Liu, Yong; Liu, Fangqi; Cai, Guoxiang; Gu, Weilie; Shen, Lijun; Zhang, Zhen

    2014-03-15

    Purpose: Ataxia telangiectasia mutated (ATM) protein is important in the DNA damage response because it repairs radiation-induced damage in cancers. We examined the effect of microRNA-223 (miR-223), a regulator of ATM expression, on radiation sensitivity of cancer cells. Methods and Materials: Human embryonic kidney 293 T (293T) cells were infected with pLL3.7-miR-223 plasmid to generate the pLL3.7-miR-223 and -empty virus (EV) lentivirus (miR-223 and EV). A dual luciferase assay in which the reporter contained wild-type 3′ untranslated region (UTR) of ATM was performed. U87MG cells were infected with miR-223 or EV to establish the overexpressed stable cell lines (U87-223 or U87-EV, respectively). Cells were irradiated in vitro, and dose enhancement ratios at 2 Gy (DER{sub 2}) were calculated. Hind legs of BALB/c athymic mice were injected with U87-223 or U87-EV cells; after 2 weeks, half of the tumors were irradiated. Tumor volumes were tracked for a total of 5 weeks. Results: The dual luciferase reporter assay showed a significant reduction in luciferase activity of 293T cells cotransfected with miR-223 and the ATM 3′UTR compared to that in EV control. Overexpression of miR-223 in U87MG cells showed that ATM expression was significantly downregulated in the U87-223 cells compared to that in U87-EV (ATM/β-actin mRNA 1.0 vs 1.5, P<.05). U87-223 cells were hypersensitive to radiation compared to U87-EV cells in vitro (DER{sub 2} = 1.32, P<.01). Mice injected with miR-223-expressing tumors had almost the same tumors after 3 weeks (1.5 cm{sup 3} vs 1.7 cm{sup 3}). However, irradiation significantly decreased tumor size in miR-223-expressing tumors compared to those in controls (0.033 cm{sup 3} vs 0.829 cm{sup 3}). Conclusions: miR-223 overexpression downregulates ATM expression and sensitizes U87 cells to radiation in vitro and in vivo. MicroRNA-223 may be a novel cancer-targeting therapy, although its cancer- and patient-specific roles are

  17. Targeting Drug-Sensitive and -Resistant Strains of Mycobacterium tuberculosis by Inhibition of Src Family Kinases Lowers Disease Burden and Pathology.

    PubMed

    Chandra, Pallavi; Rajmani, R S; Verma, Garima; Bhavesh, Neel Sarovar; Kumar, Dhiraj

    2016-01-01

    In view of emerging drug resistance among bacterial pathogens, including Mycobacterium tuberculosis, the development of novel therapeutic strategies is increasingly being sought. A recent paradigm in antituberculosis (anti-TB) drug development is to target the host molecules that are crucial for intracellular survival of the pathogen. We previously showed the importance of Src tyrosine kinases in mycobacterial pathogenesis. Here, we report that inhibition of Src significantly reduced survival of H37Rv as well as multidrug-resistant (MDR) and extremely drug-resistant (XDR) strains of M. tuberculosis in THP-1 macrophages. Src inhibition was also effective in controlling M. tuberculosis infection in guinea pigs. In guinea pigs, reduced M. tuberculosis burden due to Src inhibition also led to a marked decline in the disease pathology. In agreement with the theoretical framework of host-directed approaches against the pathogen, Src inhibition was equally effective against an XDR strain in controlling infection in guinea pigs. We propose that Src inhibitors could be developed into effective host-directed anti-TB drugs, which could be indiscriminately used against both drug-sensitive and drug-resistant strains of M. tuberculosis. IMPORTANCE The existing treatment regimen for tuberculosis (TB) suffers from deficiencies like high doses of antibiotics, long treatment duration, and inability to kill persistent populations in an efficient manner. Together, these contribute to the emergence of drug-resistant tuberculosis. Recently, several host factors were identified which help intracellular survival of Mycobacterium tuberculosis within the macrophage. These factors serve as attractive targets for developing alternate therapeutic strategies against M. tuberculosis. This strategy promises to be effective against drug-resistant strains. The approach also has potential to considerably lower the risk of emergence of new drug-resistant strains. We explored tyrosine kinase Src as a

  18. miR-223 enhances the sensitivity of non-small cell lung cancer cells to erlotinib by targeting the insulin-like growth factor-1 receptor.

    PubMed

    Zhao, Feng-Yi; Han, Jing; Chen, Xie-Wan; Wang, Jiang; Wang, Xu-Dong; Sun, Jian-Guo; Chen, Zheng-Tang

    2016-07-01

    Lung cancer is the leading cause of cancer-related fatalities worldwide, and non-small cell lung cancer (NSCLC) is the main pathological type. MicroRNAs (miRNAs or miRs) are a class of small non-coding RNAs, which are involved in tumor initiation and progression. miR‑223 is a tumor suppressor miRNA that has been reported in various types of cancer, including lung cancer. In the present study, to characterize the biological behavior of miR‑223 in NSCLC, we established an miR‑223 overexpression model in erlotinib-resistant PC‑9 (PC‑9/ER) cells by infection with lentivirus to induce the overexpression of miR‑223. As a result, miR‑223 enhanced the sensitivity of the PC‑9/ER cells to erlotinib by inducing apoptosis in vitro. Additionally, in vivo experiments were performed using nude mice which were injected with the cancer cells [either the PC‑9 (not resistant), PC‑9/ER, or the PC‑9/ER cells infected with miR‑223)]. We found that the tumor volumes were reduced in the rats injected with the cells infected with miR‑223. To further explore the underlying mechanisms, reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and western blot analysis were used to identify the target molecules of miR‑223. miR‑223 was demonstrated to act as a local regulator of insulin-like growth factor-1 receptor (IGF-1R) in the acquired resistance to tyrosine kinase inhibitors (TKIs). Notably, the οverexpression of IGF-1R in NSCLC was downregulated by miR‑223, and the activation of Akt/S6, the downstream pathway, was also inhibited. The inhibition of IGF-1R by miR‑223 was attenuated by exogenous IGF-1 expression. Therefore, miR‑223 may regulate the acquired resistance of PC‑9/ER cells to erlotinib by targeting the IGF-1R/Akt/S6 signaling pathway. The overexpression of miR‑223 may partially reverse the acquired resistance to epidermal growth factor receptor-TKIs, thus, providing a potential therapeutic strategy for TKI

  19. Development of a bone-targeted pH-sensitive liposomal formulation containing doxorubicin: physicochemical characterization, cytotoxicity, and biodistribution evaluation in a mouse model of bone metastasis

    PubMed Central

    Ferreira, Diêgo dos Santos; Faria, Samilla Dornelas; Lopes, Sávia Caldeira de Araújo; Teixeira, Cláudia Salviano; Malachias, Angelo; Magalhães-Paniago, Rogério; de Souza Filho, José Dias; Oliveira, Bruno Luis de Jesus Pinto; Guimarães, Alexander Ramos; Caravan, Peter; Ferreira, Lucas Antônio Miranda; Alves, Ricardo José; Oliveira, Mônica Cristina

    2016-01-01

    Background Despite recent advances in cancer therapy, the treatment of bone tumors remains a major challenge. A possible underlying hypothesis, limitation, and unmet need may be the inability of therapeutics to penetrate into dense bone mineral, which can lead to poor efficacy and high toxicity, due to drug uptake in healthy organs. The development of nanostructured formulations with high affinity for bone could be an interesting approach to overcome these challenges. Purpose To develop a liposomal formulation with high affinity for hydroxyapatite and the ability to release doxorubicin (DOX) in an acidic environment for future application as a tool for treatment of bone metastases. Materials and methods Liposomes were prepared by thin-film lipid hydration, followed by extrusion and the sulfate gradient-encapsulation method. Liposomes were characterized by average diameter, ζ-potential, encapsulation percentage, X-ray diffraction, and differential scanning calorimetry. Release studies in buffer (pH 7.4 or 5), plasma, and serum, as well as hydroxyapatite-affinity in vitro analysis were performed. Cytotoxicity was evaluated by MTT assay against the MDA-MB-231 cell line, and biodistribution was assessed in bone metastasis-bearing animals. Results Liposomes presented suitable diameter (~170 nm), DOX encapsulation (~2 mg/mL), controlled release, and good plasma and serum stability. The existence of interactions between DOX and the lipid bilayer was proved through differential scanning calorimetry and small-angle X-ray scattering. DOX release was faster when the pH was in the range of a tumor than at physiological pH. The bone-targeted formulation showed a strong affinity for hydroxyapatite. The encapsulation of DOX did not interfere in its intrinsic cytotoxicity against the MDA-MB-231 cell line. Biodistribution studies demonstrated high affinity of this formulation for tumors and reduction of uptake in the heart. Conclusion These results suggest that bone-targeted pH-sensitive

  20. Advancing forensic RNA typing: On non-target secretions, a nasal mucosa marker, a differential co-extraction protocol and the sensitivity of DNA and RNA profiling.

    PubMed

    van den Berge, Margreet; Bhoelai, Bryan; Harteveld, Joyce; Matai, Anuska; Sijen, Titia

    2016-01-01

    The forensic identification of human body fluids and tissues by means of messenger RNA (mRNA) profiling is a long studied methodology that is increasingly applied to casework samples. Previously, we have described an mRNA multiplex system that targets blood, saliva, semen, menstrual secretion, vaginal mucosa and skin (Lindenbergh et al. and van den Berge et al.). In this study we consider various topics to improve this mRNA profiling system or its use and adapt the method accordingly. Bodily secretions that may be encountered at a crime scene whilst not targeted by the multiplex-id est nasal mucosa, sweat, tears, faeces and urine-were examined for false positive signals. The results prompted us to identify a nasal mucosa marker that allows the discrimination of nasal mucosa from saliva or vaginal mucosa and nosebleed blood from peripheral blood. An updated version of the multiplex was prepared to which the nasal mucosa marker was added and in which markers for semen, vaginal mucosa and blood were replaced. Lactobacillus markers were regarded unsuitable as replacement for vaginal mucosa mRNA markers because of background signals on penile swabs that appeared devoid of female DNA. Furthermore, we provide approaches to deal with highly unbalanced mixtures. First, a differential extraction protocol was incorporated into a co-extraction protocol to allow DNA and RNA analysis of separated non-sperm and sperm fractions. In a second approach, besides the standard multiplex, a customized multiplex is used which excludes markers for prevailing cell types. This allows the use of lower cDNA inputs for the prevailing cell types and higher inputs for cell types that appear masked. Additionally, we assessed the relation between the percentage of alleles or markers detected in DNA or RNA profiles when decreasing sample amounts are analysed. While blood, saliva, semen and menstrual secretion show the trend that DNA profiling is more sensitive than RNA profiling, the reverse is seen

  1. Aerospace Safety Advisory Panel

    NASA Technical Reports Server (NTRS)

    1989-01-01

    This report provides findings, conclusions and recommendations regarding the National Space Transportation System (NSTS), the Space Station Freedom Program (SSFP), aeronautical projects and other areas of NASA activities. The main focus of the Aerospace Safety Advisory Panel (ASAP) during 1988 has been monitoring and advising NASA and its contractors on the Space Transportation System (STS) recovery program. NASA efforts have restored the flight program with a much better management organization, safety and quality assurance organizations, and management communication system. The NASA National Space Transportation System (NSTS) organization in conjunction with its prime contractors should be encouraged to continue development and incorporation of appropriate design and operational improvements which will further reduce risk. The data from each Shuttle flight should be used to determine if affordable design and/or operational improvements could further increase safety. The review of Critical Items (CILs), Failure Mode Effects and Analyses (FMEAs) and Hazard Analyses (HAs) after the Challenger accident has given the program a massive data base with which to establish a formal program with prioritized changes.

  2. Aerospace Safety Advisory Panel

    NASA Astrophysics Data System (ADS)

    1989-03-01

    This report provides findings, conclusions and recommendations regarding the National Space Transportation System (NSTS), the Space Station Freedom Program (SSFP), aeronautical projects and other areas of NASA activities. The main focus of the Aerospace Safety Advisory Panel (ASAP) during 1988 has been monitoring and advising NASA and its contractors on the Space Transportation System (STS) recovery program. NASA efforts have restored the flight program with a much better management organization, safety and quality assurance organizations, and management communication system. The NASA National Space Transportation System (NSTS) organization in conjunction with its prime contractors should be encouraged to continue development and incorporation of appropriate design and operational improvements which will further reduce risk. The data from each Shuttle flight should be used to determine if affordable design and/or operational improvements could further increase safety. The review of Critical Items (CILs), Failure Mode Effects and Analyses (FMEAs) and Hazard Analyses (HAs) after the Challenger accident has given the program a massive data base with which to establish a formal program with prioritized changes.

  3. Wind Advisory System

    NASA Technical Reports Server (NTRS)

    Curto, Paul A. (Inventor); Brown, Gerald E. (Inventor); Zysko, Jan A. (Inventor)

    2001-01-01

    The present invention is a two-part wind advisory system comprising a ground station at an airfield and an airborne unit placed inside an aircraft. The ground station monitors wind conditions (wind speed, wind direction, and wind gust) at the airfield and transmits the wind conditions and an airfield ID to the airborne unit. The airborne unit identifies the airfield by comparing the received airfield ID with airfield IDs stored in a database. The airborne unit also calculates the headwind and crosswind for each runway in both directions at the airfield using the received wind conditions and runway information stored in the database. The airborne unit then determines a recommended runway for takeoff and landing operations of the aircraft based on th runway having the greatest headwind value and displays the airfield ID, wind conditions, and recommended runway to the pilot. Another embodiment of the present invention includes a wireless internet based airborne unit in which the airborne unit can receive the wind conditions from the ground station over the internet.

  4. Aerospace safety advisory panel

    NASA Technical Reports Server (NTRS)

    1995-01-01

    The Aerospace Safety Advisory Panel (ASAP) monitored NASA's activities and provided feedback to the NASA Administrator, other NASA officials and Congress throughout the year. Particular attention was paid to the Space Shuttle, its launch processing and planned and potential safety improvements. The Panel monitored Space Shuttle processing at the Kennedy Space Center (KSC) and will continue to follow it as personnel reductions are implemented. There is particular concern that upgrades in hardware, software, and operations with the potential for significant risk reduction not be overlooked due to the extraordinary budget pressures facing the agency. The authorization of all of the Space Shuttle Main Engine (SSME) Block II components portends future Space Shuttle operations at lower risk levels and with greater margins for handling unplanned ascent events. Throughout the year, the Panel attempted to monitor the safety activities related to the Russian involvement in both space and aeronautics programs. This proved difficult as the working relationships between NASA and the Russians were still being defined as the year unfolded. NASA's concern for the unique safety problems inherent in a multi-national endeavor appears appropriate. Actions are underway or contemplated which should be capable of identifying and rectifying problem areas. The balance of this report presents 'Findings and Recommendations' (Section 2), 'Information in Support of Findings and Recommendations' (Section 3) and Appendices describing Panel membership, the NASA response to the March 1994 ASAP report, and a chronology of the panel's activities during the reporting period (Section 4).

  5. Using Theory to Identify Beliefs Associated with Intentions to Follow Fish Consumption Advisories Among Anglers Living in the Great Lakes Region.

    PubMed

    Niederdeppe, Jeff; Connelly, Nancy A; Labuer, T Bruce; Knuth, Barbara A

    2015-11-01

    Fish consumption advisories are issued by states, tribes, and federal agencies to provide guidance to consumers about eating sport-caught fish potentially affected by chemical contaminants. Previous work has found that while anglers report being aware that advisories are available, awareness and use of specific advisory recommendations is low. This study uses the Integrative Model of Behavioral Prediction (IMBP) to identify beliefs with potential to increase intentions to follow fish consumption advisories in Great Lakes states. We conducted a mail survey of 1,712 licensed anglers in seven of eight Great Lakes states (excluding Ohio) to gauge advisory awareness, cognitive factors influencing fish consumption behaviors (informed by the IMBP), and sociodemographic characteristics. Results show that most anglers reported being generally or vaguely aware of fish consumption advisories and try to follow them, but far fewer report being aware of specific advice needed to decide whether or not to consume different types of sport-caught fish. Informed by the IMBP, we also identify several behavioral, normative, and control beliefs that have sufficient room to change, strong associations with intentions to follow the advisories, and potential to be modified if targeted with strategic risk messages. Targeting these beliefs with strategic communication holds potential to increase the proportion of anglers intending to follow fish consumption advisory recommendations in choosing which fish to eat.

  6. 76 FR 37312 - Fruit and Vegetable Industry Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-06-27

    ...; ] DEPARTMENT OF AGRICULTURE Agricultural Marketing Service Fruit and Vegetable Industry Advisory Committee... Agriculture (USDA) Fruit and Vegetable Industry Advisory Committee and a Request for Nominations. SUMMARY: The USDA intends to reestablish the Fruit and Vegetable Industry Advisory Committee (Committee)....

  7. 77 FR 55201 - Biological and Environmental Research Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-09-07

    ... Biological and Environmental Research Advisory Committee AGENCY: Department of Energy, Office of Science... and Environmental Research Advisory Committee (BERAC). The Federal Advisory Committee Act (Pub. L. 92... Science, Office of Biological and Environmental Research, SC-23/Germantown Building, 1000...

  8. 76 FR 40768 - Occupational Information Development Advisory Panel Meeting

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-07-11

    ... ADMINISTRATION Occupational Information Development Advisory Panel Meeting AGENCY: Social Security Administration... Development Advisory Panel (panel) is a discretionary panel, established under the Federal Advisory Committee... development and testing of a content model and taxonomy, work analysis instrumentation, sampling, and...

  9. 76 FR 70804 - Occupational Information Development Advisory Panel Meeting

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-11-15

    ... ADMINISTRATION Occupational Information Development Advisory Panel Meeting AGENCY: Social Security Administration... Development Advisory Panel (panel) is a discretionary panel, established under the Federal Advisory Committee... development and testing of a content model and taxonomy, work analysis instrumentation, sampling, and...

  10. 78 FR 17640 - National Climate Assessment and Development Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-03-22

    ... National Oceanic and Atmospheric Administration National Climate Assessment and Development Advisory... the National Climate Assessment and Development Advisory Committee (NCADAC) for a six-month period.... Decker, Designated Federal Officer, National Climate Assessment and Development Advisory Committee,...

  11. 76 FR 78908 - Biological and Environmental Research Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-12-20

    ... Biological and Environmental Research Advisory Committee AGENCY: Office of Science, Department of Energy. ACTION: Notice of renewal of the Biological and Environmental Research Advisory Committee. SUMMARY... Biological and Environmental Research Advisory Committee will be renewed for a two- year period...

  12. 76 FR 31319 - Biological and Environmental Research Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-05-31

    ... Biological and Environmental Research Advisory Committee AGENCY: Office of Science, Department of Energy... the Biological and Environmental Research Advisory Committee (BERAC). The Federal Advisory Committee..., Office of Biological and Environmental Research, SC-23/Germantown Building, 1000 Independence Avenue,...

  13. 78 FR 29145 - President's National Security Telecommunications Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-05-17

    ... SECURITY President's National Security Telecommunications Advisory Committee AGENCY: National Protection... Advisory Committee Meeting; Correction. SUMMARY: The Department of Homeland Security published a document...'s National Security Telecommunications Advisory Committee (NSTAC). The document contained...

  14. 77 FR 61421 - Committee Name: Homeland Security Academic Advisory Council

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-10-09

    ... SECURITY Committee Name: Homeland Security Academic Advisory Council AGENCY: Department of Homeland Security. ACTION: Committee Management; Notice of Federal Advisory Committee Meeting. SUMMARY: The Homeland Security Academic Advisory Council (HSAAC) will meet on October 24, 2012 in Washington, DC. The...

  15. 77 FR 20643 - Blood Products Advisory Committee; Notice of Meeting

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-04-05

    ... HUMAN SERVICES Food and Drug Administration Blood Products Advisory Committee; Notice of Meeting AGENCY... public. Name of Committee: Blood Products Advisory Committee. General Function of the Committee: To... following links. Blood Products Advisory Committee Web Cast Link May 15...

  16. 76 FR 39405 - Blood Products Advisory Committee; Notice of Meeting

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-07-06

    ... HUMAN SERVICES Food and Drug Administration Blood Products Advisory Committee; Notice of Meeting AGENCY... public. Name of Committee: Blood Products Advisory Committee. General Function of the Committee: To... available at the following links. Blood Products Advisory Committee Day 1:...

  17. 75 FR 60493 - Aviation Rulemaking Advisory Committee; Renewal

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-09-30

    ... Federal Aviation Administration Aviation Rulemaking Advisory Committee; Renewal AGENCY: Federal Aviation... Regulations, the FAA gives notice it has renewed the Aviation Rulemaking Advisory Committee (ARAC) for a 2..., Executive Director, Aviation Rulemaking Advisory Committee. BILLING CODE 4910-13-P...

  18. 78 FR 20685 - Aviation Security Advisory Committee (ASAC) Meeting

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-04-05

    ... SECURITY Transportation Security Administration Aviation Security Advisory Committee (ASAC) Meeting AGENCY... Committee Meeting. SUMMARY: The Aviation Security Advisory Committee (ASAC) will meet in Arlington, VA. This.... L. 92-463). The Aviation Security Advisory Committee (ASAC) provides advice and...

  19. 77 FR 56909 - Aviation Rulemaking Advisory Committee (ARAC); Renewal

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-09-14

    ... Federal Aviation Administration Aviation Rulemaking Advisory Committee (ARAC); Renewal AGENCY: Federal Aviation Administration (FAA), DOT. ACTION: Notice of Renewal. SUMMARY: The FAA announces the charter renewal of the Aviation Rulemaking Advisory Committee (ARAC), a Federal Advisory Committee that works...

  20. 76 FR 19952 - Davy Crockett National Forest Resource Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-04-11

    ... Davy Crockett National Forest Resource Advisory Committee AGENCY: Forest Service, USDA. ACTION: Notice of Public Meeting, Davy Crockett National Forest Resource Advisory Committee. SUMMARY: In accordance.... Department of Agriculture, Forest Service, Davy Crockett National Forest Resource Advisory Committee...

  1. 76 FR 14647 - Sabine National Forest Resource Advisory Committee Meeting

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-03-17

    ...; ] DEPARTMENT OF AGRICULTURE Sabine National Forest Resource Advisory Committee Meeting AGENCY: Forest Service, USDA. ACTION: Notice of Public Meeting, Sabine National Forest Resource Advisory Committee. SUMMARY: In.... Department of Agriculture, Forest Service, Sabine National Forest Resource Advisory Committee (RAC)...

  2. 75 FR 72863 - Motor Carrier Safety Advisory Committee Public Meeting

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-11-26

    ... Federal Motor Carrier Safety Administration Motor Carrier Safety Advisory Committee Public Meeting AGENCY: Federal Motor Carrier Safety Administration, DOT. ACTION: Notice of Motor Carrier Safety Advisory Committee Meeting. SUMMARY: FMCSA announces that the Agency's Motor Carrier Safety Advisory Committee...

  3. 75 FR 50797 - Motor Carrier Safety Advisory Committee Public Meeting

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-08-17

    ... Federal Motor Carrier Safety Administration Motor Carrier Safety Advisory Committee Public Meeting AGENCY: Federal Motor Carrier Safety Administration (FMCSA), DOT. ACTION: Notice of Motor Carrier Safety Advisory Committee Meeting. SUMMARY: FMCSA announces that its Motor Carrier Safety Advisory Committee (MCSAC)...

  4. 75 FR 29384 - Motor Carrier Safety Advisory Committee Public Meeting

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-05-25

    ... Federal Motor Carrier Safety Administration Motor Carrier Safety Advisory Committee Public Meeting AGENCY: Federal Motor Carrier Safety Administration (FMCSA), DOT. ACTION: Notice of Motor Carrier Safety Advisory Committee meeting. SUMMARY: FMCSA announces that its Motor Carrier Safety Advisory Committee (MCSAC)...

  5. 78 FR 53497 - Commercial Space Transportation Advisory Committee; Closed Session

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-08-29

    ... Federal Aviation Administration Commercial Space Transportation Advisory Committee; Closed Session AGENCY: Federal Aviation Administration (FAA), DOT. ACTION: Notice of Commercial Space Transportation Advisory... closed session of the Commercial Space Transportation Advisory Committee (COMSTAC). The special...

  6. Rapid and Sensitive Detection of Pseudomonas aeruginosa in Chlorinated Water and Aerosols targeting gyrB gene using Real-time PCR

    PubMed Central

    Lee, Chang Soo; Wetzel, Kaedra; Buckley, Timothy; Wozniak, Daniel; Lee, Jiyoung

    2011-01-01

    Aims For the rapid detection of P. aeruginosa from chlorinated water and aerosols, gyrB gene-based real-time PCR assay was developed and investigated. Methods and Results Two novel primer sets (pa722F/746MGB/899R and pa722F/746MGB/788R) were designed using the most updated 611 Pseudomonas and 748 other bacterial gyrB genes for achieving high specificity. Their specificity showed 100% accuracy when tested with various strains including clinical isolates from cystic fibrosis patients. The assay was tested with P. aeruginosa-containing chlorinated water and aerosols to simulate the waterborne and airborne transmission routes (detection limit 3.3 × 102 CFU·PCR−1 − 2.3 × 103 CFU·PCR−1). No chlorine interference in real-time PCR was observed at drinking water level (~ 1 mg·L−1), but high level of chorine (12 mg·L−1) interfered the assay, thus neutralization was needed. P. aeruginosa in aerosol was successfully detected after capturing with gelatin filters with minimum 2 min of sampling time when the initial concentration of 104 CFU·mL−1 bacteria existed in the nebulizer. Conclusions A highly specific and rapid assay (2–3 hrs) was developed by targeting gyrB gene for the detection of P. aeruginosa in chlorinated water and aerosols, combined with optimized sample collection methods and sample processing, so the direct DNA extraction from either water or aerosol was possible while achieving the desired sensitivity of the method. Significance and Impact The new assay can provide timely and accurate risk assessment to prevent P. aeruginosa exposure from water and aerosol, resulting in reduced disease burden, especially among immune-compromised and susceptible individuals. This approach can be easily utilized as a platform technology for the detection of other types of microorganisms, especially for those that are transmitted via water and aerosol routes, such as Legionella pneumophila. PMID:21794031

  7. Identification of Unanticipated and Novel N-Acyl L-Homoserine Lactones (AHLs) Using a Sensitive Non-Targeted LC-MS/MS Method

    PubMed Central

    Patel, Nishaben M.; Moore, Joseph D.; Blackwell, Helen E.; Amador-Noguez, Daniel

    2016-01-01

    N-acyl L-homoserine lactones (AHLs) constitute a predominant class of quorum-sensing signaling molecules used by Gram-negative bacteria. Here, we report a sensitive and non-targeted HPLC-MS/MS method based on parallel reaction monitoring (PRM) to identify and quantitate known, unanticipated, and novel AHLs in microbial samples. Using a hybrid quadrupole-high resolution mass analyzer, this method integrates MS scans and all-ion fragmentation MS/MS scans to allow simultaneous detection of AHL parent-ion masses and generation of full mass spectra at high resolution and high mass accuracy in a single chromatographic run. We applied this method to screen for AHL production in a variety of Gram-negative bacteria (i.e. B. cepacia, E. tarda, E. carotovora, E. herbicola, P. stewartii, P. aeruginosa, P. aureofaciens, and R. sphaeroides) and discovered that nearly all of them produce a larger set of AHLs than previously reported. Furthermore, we identified production of an uncommon AHL (i.e. 3-oxo-C7-HL) in E. carotovora and P. stewartii, whose production has only been previously observed within the genera Serratia and Yersinia. Finally, we used our method to quantitate AHL degradation in B. cepacia, E. carotovora, E. herbicola, P. stewartii, P. aeruginosa, P. aureofaciens, the non-AHL producer E. coli, and the Gram-positive bacterium B. subtilis. We found that AHL degradation ability varies widely across these microbes, of which B. subtilis and E. carotovora are the best degraders, and observed that there is a general trend for AHLs containing long acyl chains (≥10 carbons) to be degraded at faster rates than AHLs with short acyl chains (≤6 carbons). PMID:27706219

  8. 78 FR 12059 - Federal Advisory Committee Act; Technological Advisory Council

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-02-21

    ...., Washington, DC 20554. FOR FURTHER INFORMATION CONTACT: Walter Johnston, Chief, Electromagnetic Compatibility Division, 202-418-0807; Walter.Johnston@FCC.gov . SUPPLEMENTARY INFORMATION: The FCC Technological Advisory.../ . The public may submit written comments before the meeting to: Walter Johnston, the FCC's...

  9. 77 FR 30289 - Federal Advisory Committee Act; Technological Advisory Council

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-05-22

    .... FOR FURTHER INFORMATION CONTACT: Walter Johnston, Chief, Electromagnetic Compatibility Division, 202-418-0807; Walter.Johnston@FCC.gov . SUPPLEMENTARY INFORMATION: The FCC Technological Advisory Council...://www.fcc.gov/live/ . The public may submit written comments before the meeting to: Walter Johnston,...

  10. 76 FR 58513 - Federal Advisory Committee Act; Technological Advisory Council

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-09-21

    ...., Washington, DC 20554. FOR FURTHER INFORMATION CONTACT: Walter Johnston, Chief, Electromagnetic Compatibility Division, 202-418-0807; Walter.Johnston@FCC.gov . SUPPLEMENTARY INFORMATION: Technical Advisory Council... public may submit written comments before the meeting to: Walter Johnston, the FCC's Designated...

  11. 77 FR 52332 - Federal Advisory Committee Act; Technological Advisory Council

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-08-29

    ..., DC 20554. FOR FURTHER INFORMATION CONTACT: Walter Johnston, Chief, Electromagnetic Compatibility Division, 202-418-0807; Walter.Johnston@FCC.gov . SUPPLEMENTARY INFORMATION: The FCC Technological Advisory... public may submit written comments before the meeting to: Walter Johnston, the FCC's Designated...

  12. 78 FR 55255 - Federal Advisory Committee Act; Technological Advisory Council

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-09-10

    ... INFORMATION CONTACT: Walter Johnston, Chief, Electromagnetic Compatibility Division, 202-418-0807; Walter.Johnston@FCC.gov . SUPPLEMENTARY INFORMATION: The FCC Technological Advisory Council will discuss progress...://www.fcc.gov/live/ . The public may submit written comments before the meeting to: Walter Johnston,...

  13. 77 FR 70543 - Pipeline Safety: Meeting of the Gas Pipeline Advisory Committee and the Liquid Pipeline Advisory...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-11-26

    ... Pipeline Advisory Committee and the Liquid Pipeline Advisory Committee AGENCY: Pipeline and Hazardous... Technical Pipeline Safety Standards Committee, and the Liquid Pipeline Advisory Committee (LPAC), also known as the Technical Hazardous Liquid Pipeline Safety Standards Committee. The committees will meet...

  14. 75 FR 7440 - Eastern Washington Cascades Provincial Advisory Committee and the Yakima Provincial Advisory...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-02-19

    ... Forest Service Eastern Washington Cascades Provincial Advisory Committee and the Yakima Provincial Advisory Committee AGENCY: Forest Service, USDA. ACTION: Notice of meeting. SUMMARY: The Eastern Washington..., 2010 at the Sunnyslope Fire Station, 206 Easy Street, Wenatchee, WA. During this meeting...

  15. 75 FR 60484 - NASA Advisory Council; Science Committee; Earth Science Subcommittee; Applied Sciences Advisory...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-09-30

    ... SPACE ADMINISTRATION NASA Advisory Council; Science Committee; Earth Science Subcommittee; Applied Sciences Advisory Group Meeting AGENCY: National Aeronautics and Space Administration. ACTION: Notice of..., the National Aeronautics and Space Administration (NASA) announces a meeting of the Applied...

  16. 78 FR 9917 - The President's Management Advisory Board (PMAB); Notification of Upcoming Public Advisory Meeting

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-02-12

    ... effective strategies for the implementation of best business practices to improve Federal Government... ADMINISTRATION The President's Management Advisory Board (PMAB); Notification of Upcoming Public Advisory Meeting AGENCY: Office of Executive Councils, U.S. General Services Administration (GSA). ACTION: Meeting...

  17. 76 FR 65511 - The President's Management Advisory Board (PMAB); Notification of Upcoming Public Advisory Meeting

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-10-21

    ... effective strategies for the implementation of best business practices to improve Federal Government... ADMINISTRATION The President's Management Advisory Board (PMAB); Notification of Upcoming Public Advisory Meeting AGENCY: Office of Executive Councils, U.S. General Services Administration (GSA). ACTION: Meeting...

  18. 78 FR 53127 - Delta-Bienville Resource Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-08-28

    ...; ] DEPARTMENT OF AGRICULTURE Forest Service Delta-Bienville Resource Advisory Committee AGENCY: Forest Service, USDA. ACTION: Notice of meeting. Date change. SUMMARY: The Delta-Bienville Resource Advisory...

  19. 76 FR 49431 - Missouri River Resource Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-08-10

    ...; ] DEPARTMENT OF AGRICULTURE Forest Service Missouri River Resource Advisory Committee AGENCY: Forest Service, USDA. ACTION: Notice of meeting. SUMMARY: The Missouri River Resource Advisory Committee will meet...

  20. 76 FR 28415 - Fresno County Resource Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-05-17

    ... Robbin Ekman, Fresno County Resource Advisory Committee Coordinator, c/ o Sierra National Forest, High... FURTHER INFORMATION CONTACT: Robbin Ekman, Fresno County Resource ] Advisory Committee Coordinator,...

  1. 76 FR 44302 - Fresno County Resource Advisory Committee

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-07-25

    ... Robbin Ekman, Fresno County Resource Advisory Committee Coordinator, c/ o Sierra National Forest, High... FURTHER INFORMATION CONTACT: Robbin Ekman, Fresno County Resource Advisory Committee Coordinator,...

  2. 36 CFR 219.18 - Role of advisory committees.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... address an issue. Each Forest or Grassland Supervisor must have access to an advisory committee with... forest or grassland. Responsible officials may request establishment of advisory committees and...

  3. 36 CFR 219.18 - Role of advisory committees.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... address an issue. Each Forest or Grassland Supervisor must have access to an advisory committee with... forest or grassland. Responsible officials may request establishment of advisory committees and...

  4. 16 CFR 16.4 - Advisory Committee Management Officer.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... designate the Executive Director as the Advisory Committee Management Officer who shall: (1) Exercise... Commission's advisory committees; (5) Maintain information on the nature, functions, and operations of...

  5. Senior Adult Consumer Advisory Manual,

    ERIC Educational Resources Information Center

    Ater, E. Carolyn, Ed.

    This manual is intended for use by senior adult peer advisors (age 60 and over) engaged in helping relationships in providing consumer education to other senior adults. The advisory procedures are based on a problem solving approach which incorporates the development of a self-help concept. Chapter 1 provides information on consumer advising. It…

  6. Objectives of Youth Advisory Services.

    ERIC Educational Resources Information Center

    World Health Organization, Copenhagen (Denmark). Regional Office for Europe.

    Professionals concerned with youth advisory services from 10 countries met to exchange appropriate experiences and information on their role, organizations and operations, especially within the framework of other community services. Changes in their roles and objectives received special attention to determine possibilities for prevention and early…

  7. Advisory List of Computer Courseware.

    ERIC Educational Resources Information Center

    North Carolina State Dept. of Public Instruction, Raleigh. Div. of Educational Media.

    Computer courseware appropriate for instruction in grades K-12 is listed in two advisory lists. Entries, selected from materials submitted by producers which received favorable reviews by educators, are arranged in the following categories: arts education, communication skills, mathematics, science, utility (a quiz generator), word processing, and…

  8. Serving Boys through Readers' Advisory

    ERIC Educational Resources Information Center

    Sullivan, Michael

    2010-01-01

    Based on more than twenty years' experience working to get boys interested in reading, the author now offers his first readers' advisory volume. With an emphasis on nonfiction and the boy-friendly categories of genre fiction, the work offers a wealth of material including: (1) Suggestions for how to booktalk one-on-one as well as in large groups;…

  9. Writing Reviews for Readers' Advisory

    ERIC Educational Resources Information Center

    Hooper, Brad

    2010-01-01

    Reviews are an important resource for readers' advisory and collection development. They are also a helpful promotional tool, introducing patrons to what is new on the shelf. This resource includes: (1) Tips for writing strong, relevant reviews; (2) Different ways reviews can be used to promote your library; and (3) A chapter by Joyce Saricks…

  10. Aerospace Safety Advisory Panel

    NASA Technical Reports Server (NTRS)

    2001-01-01

    This annual report is based on the activities of the Aerospace Safety Advisory Panel in calendar year 2000. During this year, the construction of the International Space Station (ISS) moved into high gear. The launch of the Russian Service Module was followed by three Space Shuttle construction and logistics flights and the deployment of the Expedition One crew. Continuous habitation of the ISS has begun. To date, both the ISS and Space Shuttle programs have met or exceeded most of their flight objectives. In spite of the intensity of these efforts, it is clear that safety was always placed ahead of cost and schedule. This safety consciousness permitted the Panel to devote more of its efforts to examining the long-term picture. With ISS construction accelerating, demands on the Space Shuttle will increase. While Russian Soyuz and Progress spacecraft will make some flights, the Space Shuttle remains the primary vehicle to sustain the ISS and all other U.S. activities that require humans in space. Development of a next generation, human-rated vehicle has slowed due to a variety of technological problems and the absence of an approach that can accomplish the task significantly better than the Space Shuttle. Moreover, even if a viable design were currently available, the realities of funding and development cycles suggest that it would take many years to bring it to fruition. Thus, it is inescapable that for the foreseeable future the Space Shuttle will be the only human-rated vehicle available to the U.S. space program for support of the ISS and other missions requiring humans. Use of the Space Shuttle will extend well beyond current planning, and is likely to continue for the life of the ISS.

  11. Aerospace Safety Advisory Panel

    NASA Technical Reports Server (NTRS)

    1999-01-01

    This report covers the activities of the Aerospace Safety Advisory Panel (ASAP) for calendar year 1998-a year of sharp contrasts and significant successes at NASA. The year opened with the announcement of large workforce cutbacks. The slip in the schedule for launching the International Space Station (ISS) created a 5-month hiatus in Space Shuttle launches. This slack period ended with the successful and highly publicized launch of the STS-95 mission. As the year closed, ISS assembly began with the successful orbiting and joining of the Functional Cargo Block (FGB), Zarya, from Russia and the Unity Node from the United States. Throughout the year, the Panel maintained its scrutiny of NASAs safety processes. Of particular interest were the potential effects on safety of workforce reductions and the continued transition of functions to the Space Flight Operations Contractor. Attention was also given to the risk management plans of the Aero-Space Technology programs, including the X-33, X-34, and X-38. Overall, the Panel concluded that safety is well served for the present. The picture is not as clear for the future. Cutbacks have limited the depth of talent available. In many cases, technical specialties are "one deep." The extended hiring freeze has resulted in an older workforce that will inevitably suffer significant departures from retirements in the near future. The resulting "brain drain" could represent a future safety risk unless appropriate succession planning is started expeditiously. This and other topics are covered in the section addressing workforce. In the case of the Space Shuttle, beneficial and mandatory safety and operational upgrades are being delayed because of a lack of sufficient present funding. Likewise, the ISS has little flexibility to begin long lead-time items for upgrades or contingency planning.

  12. 75 FR 60458 - Federal Advisory Committee Act; Advisory Committee on Diversity for Communications in the Digital...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-09-30

    ... COMMISSION Federal Advisory Committee Act; Advisory Committee on Diversity for Communications in the Digital... Communications Commission's (FCC) Advisory Committee on Diversity for Communications in the Digital Age (``Diversity Committee'') will hold a meeting on Thursday, October 14, 2010 at 2 p.m. at the...

  13. 75 FR 20844 - Federal Advisory Committee Act; Advisory Committee on Diversity for Communications in the Digital...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-04-21

    ... COMMISSION Federal Advisory Committee Act; Advisory Committee on Diversity for Communications in the Digital... Communications Commission's (FCC) Advisory Committee on Diversity for Communications in the Digital Age (``Diversity Committee'') will hold a meeting on Tuesday, June 15, 2010 at 2:00 p.m. in the Commission...

  14. 75 FR 6031 - Federal Advisory Committee Act; Advisory Committee on Diversity for Communications in the Digital...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-02-05

    ... COMMISSION Federal Advisory Committee Act; Advisory Committee on Diversity for Communications in the Digital... Communications Commission's (FCC) Advisory Committee on Diversity for Communications in the Digital Age (``Diversity Committee'') will hold a meeting on Wednesday, March 24, 2010 at 2 p.m. in the Commission...

  15. 75 FR 70004 - Federal Advisory Committee Act; Advisory Committee on Diversity for Communications in the Digital...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-11-16

    ... COMMISSION Federal Advisory Committee Act; Advisory Committee on Diversity for Communications in the Digital... Communications Commission's (FCC) Advisory Committee on Diversity for Communications in the Digital Age (``Diversity Committee'') will hold a meeting on Thursday, December 2, 2010 at 2 p.m. in the Commission...

  16. 76 FR 5160 - Federal Advisory Committee Act; Advisory Committee on Diversity for Communications in the Digital...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-01-28

    ... COMMISSION Federal Advisory Committee Act; Advisory Committee on Diversity for Communications in the Digital... Communications Commission (FCC) has renewed the charter of the Advisory Committee on Diversity for Communications in the Digital Age (``Diversity Committee''). ADDRESSES: A copy of the charter is available at...

  17. 75 FR 22751 - Federal Advisory Committee; Defense Advisory Committee on Military Personnel Testing; Charter...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-04-30

    ... members, who are eminent authorities in the fields of educational and psychological testing. Committee... of the Secretary Federal Advisory Committee; Defense Advisory Committee on Military Personnel Testing... notice that it is renewing the charter for the Defense Advisory Committee on Military Personnel...

  18. 78 FR 21349 - Defense Advisory Committee on Military Personnel Testing; Notice of Federal Advisory Committee...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-04-10

    ... of the Secretary Defense Advisory Committee on Military Personnel Testing; Notice of Federal Advisory... Defense Advisory Committee on Military Personnel Testing will take place. DATES: Thursday, May 9, 2013... progress in developing computerized tests for military enlistment. Agenda: The agenda includes an...

  19. 75 FR 37410 - Federal Advisory Committee; Defense Advisory Committee on Military Personnel Testing

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-06-29

    ... From the Federal Register Online via the Government Publishing Office ] DEPARTMENT OF DEFENSE Office of the Secretary Federal Advisory Committee; Defense Advisory Committee on Military Personnel...D announces that the Defense Advisory Committee on Military Personnel Testing will meet on July...

  20. 75 FR 63823 - Federal Advisory Committee; Defense Advisory Committee on Military Personnel Testing

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-10-18

    ... From the Federal Register Online via the Government Publishing Office DEPARTMENT OF DEFENSE Office of the Secretary Federal Advisory Committee; Defense Advisory Committee on Military Personnel Testing... Defense announces that the Defense Advisory Committee on Military Personnel Testing will meet November...

  1. 75 FR 22560 - Federal Advisory Committee; U.S. Air Force Scientific Advisory Board; Charter Renewal

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-04-29

    ... of the Secretary Federal Advisory Committee; U.S. Air Force Scientific Advisory Board; Charter... that it is renewing the charter for the U.S. Air Force Scientific Advisory Board (hereafter referred to... of science and technology relating to the Air Force mission. The Board shall: a. Provide...

  2. 75 FR 25212 - Federal Advisory Committee; Defense Audit Advisory Committee (DAAC)

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-05-07

    ... of the Secretary Federal Advisory Committee; Defense Audit Advisory Committee (DAAC) AGENCY: Under... that the Defense Audit Advisory Committee will meet on May 17, 2010. Subject to the availability of... include financial reporting processes, systems of internal controls, audit processes, and processes...

  3. 78 FR 14299 - Notification of a Public Meeting of the Science Advisory Board Chemical Assessment Advisory...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-03-05

    ... AGENCY Notification of a Public Meeting of the Science Advisory Board Chemical Assessment Advisory... SAB Chemical Assessment Advisory Committee (CAAC) to receive a briefing about the Integrated Risk... for chemicals. DATES: The SAB CAAC meeting dates are Tuesday April 2, 2013 from 9 a.m. to 5:00...

  4. 75 FR 43493 - Office of the Secretary; Federal Advisory Committee; Defense Intelligence Agency Advisory Board...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-07-26

    ... of the Secretary; Federal Advisory Committee; Defense Intelligence Agency Advisory Board; Closed Meeting AGENCY: Defense Intelligence Agency, DOD. ACTION: Meeting notice. SUMMARY: Under the provisions of... that Defense Intelligence Agency Advisory Board will meet on September 1 and 2, 2010. The meeting...

  5. 75 FR 81245 - Federal Advisory Committee; Defense Intelligence Agency Advisory Board; Closed Meeting

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-12-27

    ... of the Secretary Federal Advisory Committee; Defense Intelligence Agency Advisory Board; Closed Meeting AGENCY: Defense Intelligence Agency, DoD. ACTION: Notice. SUMMARY: Under the provisions of the... Defense Intelligence Agency Advisory Board and two of its subcommittees will meet on January 26 and...

  6. 77 FR 18797 - Federal Advisory Committee; Defense Intelligence Agency (DIA) Advisory Board; Closed Meeting

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-03-28

    ... of the Secretary Federal Advisory Committee; Defense Intelligence Agency (DIA) Advisory Board; Closed... discussions of classified information relating to DIA's intelligence operations including its support to... intelligence operations. Agenda May 2, 2012 S 8:30 a.m Convene Advisory Board Meeting Mr. William...

  7. 76 FR 53671 - Federal Advisory Committee; Defense Intelligence Agency Advisory Board; Closed Meeting

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-08-29

    ... of the Secretary Federal Advisory Committee; Defense Intelligence Agency Advisory Board; Closed Meeting AGENCY: Defense Intelligence Agency (DIA), Department of Defense (DoD). ACTION: Meeting notice... Department of Defense announces that Defense Intelligence Agency Advisory Board and two of its...

  8. 76 FR 41220 - Federal Advisory Committee; Defense Intelligence Agency Advisory Board; Closed Meeting

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-07-13

    ... of the Secretary Federal Advisory Committee; Defense Intelligence Agency Advisory Board; Closed Meeting AGENCY: Defense Intelligence Agency, DoD. ACTION: Meeting notice. SUMMARY: Under the provisions of... that Defense Intelligence Agency Advisory Board and two of its subcommittees will meet on August 4...

  9. 75 FR 24926 - Federal Advisory Committee; Defense Intelligence Agency Advisory Board; Closed Meeting

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-05-06

    ... of the Secretary Federal Advisory Committee; Defense Intelligence Agency Advisory Board; Closed Meeting AGENCY: Defense Intelligence Agency, DoD. ACTION: Meeting notice. SUMMARY: Under the provisions of... that Defense Intelligence Agency Advisory Board, and its subcommittees, will meet on June 15 and...

  10. 77 FR 26734 - Forestry Research Advisory Council

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-05-07

    ... Daina Apple, Designated Federal Officer, Forestry Research Advisory Council, USDA Forest Service... proposed agenda items to (202) 205- 1530. FOR FURTHER INFORMATION CONTACT: Daina Apple, Forest...

  11. Functions and Responsibilities of Community Advisory Councils

    ERIC Educational Resources Information Center

    Cox, James C.

    1974-01-01

    Suggests that the following five functions are common to all community advisory councils: fact finding, planning, coordination and communications, activation of new resources, and evaluation. (Author/DN)

  12. 75 FR 7255 - Advisory Committee Meeting Notice

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-02-18

    ... gather information from students, staff and faculty. General deliberations leading to provisional findings for referral to the Army Education Advisory Committee will follow on 10 March beginning at...

  13. 78 FR 33049 - Advisory Committee Meeting

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-06-03

    ... Federal Grain Inspection Service operations-market overview, international programs, moisture meter... orally address the Advisory Committee. The meeting will be open to the public. Persons with...

  14. Model-based global sensitivity analysis as applied to identification of anti-cancer drug targets and biomarkers of drug resistance in the ErbB2/3 network

    PubMed Central

    Lebedeva, Galina; Sorokin, Anatoly; Faratian, Dana; Mullen, Peter; Goltsov, Alexey; Langdon, Simon P.; Harrison, David J.; Goryanin, Igor

    2012-01-01

    High levels of variability in cancer-related cellular signalling networks and a lack of parameter identifiability in large-scale network models hamper translation of the results of modelling studies into the process of anti-cancer drug development. Recently global sensitivity analysis (GSA) has been recognised as a useful technique, capable of addressing the uncertainty of the model parameters and generating valid predictions on parametric sensitivities. Here we propose a novel implementation of model-based GSA specially designed to explore how multi-parametric network perturbations affect signal propagation through cancer-related networks. We use area-under-the-curve for time course of changes in phosphorylation of proteins as a characteristic for sensitivity analysis and rank network parameters with regard to their impact on the level of key cancer-related outputs, separating strong inhibitory from stimulatory effects. This allows interpretation of the results in terms which can incorporate the effects of potential anti-cancer drugs on targets and the associated biological markers of cancer. To illustrate the method we applied it to an ErbB signalling network model and explored the sensitivity profile of its key model readout, phosphorylated Akt, in the absence and presence of the ErbB2 inhibitor pertuzumab. The method successfully identified the parameters associated with elevation or suppression of Akt phosphorylation in the ErbB2/3 network. From analysis and comparison of the sensitivity profiles of pAkt in the absence and presence of targeted drugs we derived predictions of drug targets, cancer-related biomarkers and generated hypotheses for combinatorial therapy. Several key predictions have been confirmed in experiments using human ovarian carcinoma cell lines. We also compared GSA-derived predictions with the results of local sensitivity analysis and discuss the applicability of both methods. We propose that the developed GSA procedure can serve as a

  15. Aerospace Safety Advisory Panel

    NASA Technical Reports Server (NTRS)

    2002-01-01

    This Annual Report of the Aerospace Safety Advisory Panel (ASAP) presents results of activities during calendar year 2001. The year was marked by significant achievements in the Space Shuttle and International Space Station (ISS) programs and encouraging accomplishments by the Aerospace Technology Enterprise. Unfortunately, there were also disquieting mishaps with the X-43, a LearJet, and a wind tunnel. Each mishap was analyzed in an orderly process to ascertain causes and derive lessons learned. Both these accomplishments and the responses to the mishaps led the Panel to conclude that safety and risk management is currently being well served within NASA. NASA's operations evidence high levels of safety consciousness and sincere efforts to place safety foremost. Nevertheless, the Panel's safety concerns have never been greater. This dichotomy has arisen because the focus of most NASA programs has been directed toward program survival rather than effective life cycle planning. Last year's Annual Report focused on the need for NASA to adopt a realistically long planning horizon for the aging Space Shuttle so that safety would not erode. NASA's response to the report concurred with this finding. Nevertheless, there has been a greater emphasis on current operations to the apparent detriment of long-term planning. Budget cutbacks and shifts in priorities have severely limited the resources available to the Space Shuttle and ISS for application to risk-reduction and life-extension efforts. As a result, funds originally intended for long-term safety-related activities have been used for operations. Thus, while safety continues to be well served at present, the basis for future safety has eroded. Section II of this report develops this theme in more detail and presents several important, overarching findings and recommendations that apply to many if not all of NASA's programs. Section III of the report presents other significant findings, recommendations and supporting

  16. Aerospace Safety Advisory Panel

    NASA Technical Reports Server (NTRS)

    1998-01-01

    During 1997, the Aerospace Safety Advisory Panel (ASAP) continued its safety reviews of NASA's human space flight and aeronautics programs. Efforts were focused on those areas that the Panel believed held the greatest potential to impact safety. Continuing safe Space Shuttle operations and progress in the manufacture and testing of primary components for the International Space Station (ISS) were noteworthy. The Panel has continued to monitor the safety implications of the transition of Space Shuttle operations to the United Space Alliance (USA). One area being watched closely relates to the staffing levels and skill mix in both NASA and USA. Therefore, a section of this report is devoted to personnel and other related issues that are a result of this change in NASA's way of doing business for the Space Shuttle. Attention will continue to be paid to this important topic in subsequent reports. Even though the Panel's activities for 1997 were extensive, fewer specific recommendations were formulated than has been the case in recent years. This is indicative of the current generally good state of safety of NASA programs. The Panel does, however, have several longer term concerns that have yet to develop to the level of a specific recommendation. These are covered in the introductory material for each topic area in Section 11. In another departure from past submissions, this report does not contain individual findings and recommendations for the aeronautics programs. While the Panel devoted its usual efforts to examining NASA's aeronautic centers and programs, no specific recommendations were identified for inclusion in this report. In lieu of recommendations, a summary of the Panel's observations of NASA's safety efforts in aeronautics and future Panel areas of emphasis is provided. With profound sadness the Panel notes the passing of our Chairman, Paul M. Johnstone, on December 17, 1997, and our Staff Assistant, Ms. Patricia M. Harman, on October 5, 1997. Other

  17. Aerospace Safety Advisory Panel

    NASA Technical Reports Server (NTRS)

    2002-01-01

    This report presents the results of the Aerospace Safety Advisory Panel (ASAP) activities during 2002. The format of the report has been modified to capture a long-term perspective. Section II is new and highlights the Panel's view of NASA's safety progress during the year. Section III contains the pivotal safety issues facing NASA in the coming year. Section IV includes the program area findings and recommendations. The Panel has been asked by the Administrator to perform several special studies this year, and the resulting white papers appear in Appendix C. The year has been filled with significant achievements for NASA in both successful Space Shuttle operations and International Space Station (ISS) construction. Throughout the year, safety has been first and foremost in spite of many changes throughout the Agency. The relocation of the Orbiter Major Modifications (OMMs) from California to Kennedy Space Center (KSC) appears very successful. The transition of responsibilities for program management of the Space Shuttle and ISS programs from Johnson Space Center (JSC) to NASA Headquarters went smoothly. The decision to extend the life of the Space Shuttle as the primary NASA vehicle for access to space is viewed by the Panel as a prudent one. With the appropriate investments in safety improvements, in maintenance, in preserving appropriate inventories of spare parts, and in infrastructure, the Space Shuttle can provide safe and reliable support for the ISS for the foreseeable future. Indications of an aging Space Shuttle fleet occurred on more than one occasion this year. Several flaws went undetected in the early prelaunch tests and inspections. In all but one case, the problems were found prior to launch. These incidents were all handled properly and with safety as the guiding principle. Indeed, launches were postponed until the problems were fully understood and mitigating action could be taken. These incidents do, however, indicate the need to analyze the

  18. Systems pharmacology of the nerve growth factor pathway: use of a systems biology model for the identification of key drug targets using sensitivity analysis and the integration of physiology and pharmacology

    PubMed Central

    Benson, Neil; Matsuura, Tomomi; Smirnov, Sergey; Demin, Oleg; Jones, Hannah M.; Dua, Pinky; van der Graaf, Piet H.

    2013-01-01

    The nerve growth factor (NGF) pathway is of great interest as a potential source of drug targets, for example in the management of certain types of pain. However, selecting targets from this pathway either by intuition or by non-contextual measures is likely to be challenging. An alternative approach is to construct a mathematical model of the system and via sensitivity analysis rank order the targets in the known pathway, with respect to an endpoint such as the diphosphorylated extracellular signal-regulated kinase concentration in the nucleus. Using the published literature, a model was created and, via sensitivity analysis, it was concluded that, after NGF itself, tropomyosin receptor kinase A (TrkA) was one of the most sensitive druggable targets. This initial model was subsequently used to develop a further model incorporating physiological and pharmacological parameters. This allowed the exploration of the characteristics required for a successful hypothetical TrkA inhibitor. Using these systems models, we were able to identify candidates for the optimal drug targets in the known pathway. These conclusions were consistent with clinical and human genetic data. We also found that incorporating appropriate physiological context was essential to drawing accurate conclusions about important parameters such as the drug dose required to give pathway inhibition. Furthermore, the importance of the concentration of key reactants such as TrkA kinase means that appropriate contextual data are required before clear conclusions can be drawn. Such models could be of great utility in selecting optimal targets and in the clinical evaluation of novel drugs. PMID:24427523

  19. Amifostine-conjugated pH-sensitive calcium phosphate-covered magnetic-amphiphilic gelatin nanoparticles for controlled intracellular dual drug release for dual-targeting in HER-2-overexpressing breast cancer.

    PubMed

    Li, Wei-Ming; Chiang, Chih-Sheng; Huang, Wei-Chen; Su, Chia-Wei; Chiang, Min-Yu; Chen, Jian-Yi; Chen, San-Yuan

    2015-12-28

    We developed a surfactant-free method utilizing amifostine to stably link a targeting ligand (Herceptin) to amphiphilic gelatin (AG)-iron oxide@calcium phosphate (CaP) nanoparticles with hydrophobic curcumin (CUR) and hydrophilic doxorubicin (DOX) encapsulated in the AG core and CaP shell (AGIO@CaP-CD), respectively. This multi-functional nanoparticle system has a pH-sensitive CaP shell and degradable amphiphilic gelatin (AG) core, which enables controllable sequential release of the two drugs. The dual-targeting system of AGIO@CaP-CD (HER-AGIO@CaP-CD) with a bioligand and magnetic targeting resulted in significantly elevated cellular uptake in HER2-overexpressing SKBr3 cells and more efficacious therapy than delivery of targeting ligand alone due to the synergistic cell multi-drug resistance/apoptosis-inducing effect of the CUR and DOX combination. This nanoparticle combined with Herceptin and iron oxide nanoparticles not only provided a dual-targeting functionality, but also encapsulated CUR and DOX as a dual-drug delivery system for the combination therapy. This study further demonstrated that the therapeutic efficacy of this dual-targeting co-delivery system can be improved by modifying the application duration of magnetic targeting, which makes this combination therapy system a powerful new tool for in vitro/in vivo cancer therapy, especially for HER2-positive cancers.

  20. 10 CFR 7.13 - Minutes of advisory committee meetings.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 10 Energy 1 2010-01-01 2010-01-01 false Minutes of advisory committee meetings. 7.13 Section 7.13 Energy NUCLEAR REGULATORY COMMISSION ADVISORY COMMITTEES § 7.13 Minutes of advisory committee meetings. (a) Detailed minutes shall be kept of each NRC advisory committee meeting. The minutes shall...