A Return to Love in William James and Jean-Luc Marion

ERIC Educational Resources Information Center

Rocha, Samuel

2009-01-01

In this essay Samuel Rocha primarily addresses, and challenges, the modern conception of reason and the lowly place of intuition, feeling, and love in what has become traditional philosophy and education. Drawing upon the rich thought of William James and Jean-Luc Marion, Rocha introduces the reader to a certain harmony between their ideas, most…

Comparison of red-shifted firefly luciferase Ppy RE9 and conventional Luc2 as bioluminescence imaging reporter genes for in vivo imaging of stem cells

NASA Astrophysics Data System (ADS)

Liang, Yajie; Walczak, Piotr; Bulte, Jeff W. M.

2012-01-01

One critical issue for noninvasive imaging of transplanted bioluminescent cells is the large amount of light absorption in tissue when emission wavelengths below 600 nm are used. Luciferase with a red-shifted spectrum can potentially bypass this limitation. We assessed and compared a mutant of firefly luciferase (Ppy RE9, PRE9) against the yellow luciferase luc2 gene for use in cell transplantation studies. C17.2 neural stem cells expressing PRE9-Venus and luc2-Venus were sorted by flow cytometry and assessed for bioluminescence in vitro in culture and in vivo after transplantation into the brain of immunodeficient Rag2-/- mice. We found that the luminescence from PRE9 was stable, with a peak emission at 620 nm, shifted to the red compared to that of luc2. The emission peak for PRE9 was pH-independent, in contrast to luc2, and much less affected by tissue absorbance compared to that of luc2. However, the total emitted light radiance from PRE9 was substantially lower than that of luc2, both in vitro and in vivo. We conclude that PRE9 has favorable properties as compared to luc2 in terms of pH independence, red-shifted spectrum, tissue light penetration, and signal quantification, justifying further optimization of protein expression and enzymatic activity.

Fluorophore-NanoLuc BRET Reporters Enable Sensitive In Vivo Optical Imaging and Flow Cytometry for Monitoring Tumorigenesis

PubMed Central

Schaub, Franz X; Reza, Md Shamim; Flaveny, Colin A; Li, Weimin; Musicant, Adele M; Hoxha, Sany; Guo, Min; Cleveland, John L; Amelio, Antonio L

2015-01-01

Fluorescent proteins are widely used to study molecular and cellular events, yet this traditionally relies on delivery of excitation light, which can trigger autofluorescence, photoxicity, and photobleaching, impairing their use in vivo. Accordingly, chemiluminescent light sources such as those generated by luciferases have emerged, as they do not require excitation light. However, current luciferase reporters lack the brightness needed to visualize events in deep tissues. We report the creation of chimeric eGFP-NanoLuc (GpNLuc) and LSSmOrange-NanoLuc (OgNLuc) fusion reporter proteins coined LumiFluors, which combine the benefits of eGFP or LSSmOrange fluorescent proteins with the bright, glow-type bioluminescent light generated by an enhanced small luciferase subunit (NanoLuc) of the deep sea shrimp Oplophorus gracilirostris. The intramolecular bioluminescence resonance energy transfer (BRET) that occurs between NanoLuc and the fused fluorophore generates the brightest bioluminescent signal known to date, including improved intensity, sensitivity and durable spectral properties, thereby dramatically reducing image acquisition times and permitting highly sensitive in vivo imaging. Notably, the self-illuminating and bi-functional nature of these LumiFluor reporters enables greatly improved spatio-temporal monitoring of very small numbers of tumor cells via in vivo optical imaging and also allows the isolation and analyses of single cells by flow cytometry. Thus, LumiFluor reporters are inexpensive, robust, non-invasive tools that allow for markedly improved in vivo optical imaging of tumorigenic processes. PMID:26424696

Monitoring of tumor growth and metastasis potential in MDA-MB-435s/ tk-luc human breast cancer xenografts

NASA Astrophysics Data System (ADS)

Chang, Ya-Fang; Lin, Yi-Yu; Wang, Hsin-Ell; Liu, Ren-Shen; Pang, Fei; Hwang, Jeng-Jong

2007-02-01

Molecular imaging of reporter gene expression provides a rapid, sensitive and non-invasive monitoring of tumor behaviors. In this study, we reported the establishment of a novel animal model for longitudinal examination of tumor growth kinetics and metastatic spreading in vivo. The highly metastatic human breast carcinoma MDA-MB-435s cell line was engineered to stably express herpes simplex virus type 1 thymidine kinase (HSV-1- tk) and luciferase ( luc). Both 131I-FIAU and D-luciferin were used as reporter probes. For orthotopic tumor formation, MDA-MB-435s/ tk-luc cells were implanted into the first nipple of 6-week-old female NOD/SCID mice. For metastatic study, cells were injected via the lateral tail vein. Mice-bearing MDA-MB-435s/ tk-luc tumors were scanned for tumor growth and metastatsis using Xenogen IVIS50 system. Gamma scintigraphy and whole-body autoradiography were also applied to confirm the tumor localization. The results of bioluminescence imaging as well as histopathological finding showed that tumors could be detected in femur, spine, ovary, lungs, kidney, adrenal gland, lymph nodes and muscle at 16 weeks post i.v. injection, and correlated photons could be quantified. This MDA-MB-435s/ tk-luc human breast carcinoma-bearing mouse model combined with multimodalities of molecular imaging may facilitate studies on the molecular mechanisms of cancer invasion and metastasis.

FLaapLUC: A pipeline for the generation of prompt alerts on transient Fermi-LAT γ-ray sources

NASA Astrophysics Data System (ADS)

Lenain, J.-P.

2018-01-01

The large majority of high energy sources detected with Fermi-LAT are blazars, which are known to be very variable sources. High cadence long-term monitoring simultaneously at different wavelengths being prohibitive, the study of their transient activities can help shedding light on our understanding of these objects. The early detection of such potentially fast transient events is the key for triggering follow-up observations at other wavelengths. A Python tool, FLaapLUC, built on top of the Science Tools provided by the Fermi Science Support Center and the Fermi-LAT collaboration, has been developed using a simple aperture photometry approach. This tool can effectively detect relative flux variations in a set of predefined sources and alert potential users. Such alerts can then be used to trigger target of opportunity observations with other facilities. It is shown that FLaapLUC is an efficient tool to reveal transient events in Fermi-LAT data, providing quick results which can be used to promptly organise follow-up observations. Results from this simple aperture photometry method are also compared to full likelihood analyses. The FLaapLUC package is made available on GitHub and is open to contributions by the community.

The Enunciation of the Subject: Sharing Jean-Luc Nancy's Singular Plural in the Classroom

ERIC Educational Resources Information Center

Collins, Ashok

2015-01-01

This article seeks to explore the implications of Jean-Luc Nancy's reading of the subject for educational philosophy by connecting his re-interpretation of Descartes to his later thinking on what he names the ontological singular plural. Nancy's re-imagining of the Cogito coalesces around the figure of the mouth ("la bouche") through…

Imaging of bioluminescent LNCaP-luc-M6 tumors: a new animal model for the study of metastatic human prostate cancer.

PubMed

Scatena, Caroline D; Hepner, Mischa A; Oei, Yoko A; Dusich, Joan M; Yu, Shang-Fan; Purchio, Tony; Contag, Pamela R; Jenkins, Darlene E

2004-05-15

Animal experiments examining hormone-sensitive metastatic prostate cancer using the human LNCaP cell line have been limited to endpoint analyses. To permit longitudinal studies, we generated a luciferase-expressing cell line and used bioluminescent imaging (BLI) to non-invasively monitor the in vivo growth of primary LNCaP tumors and metastasis. LNCaP.FGC cells were transfected to constitutively express firefly luciferase. LNCaP-luc-M6 cells were tested for bioluminescent signal intensity and hormone responsiveness in vitro. The cells were implanted in subcutaneous and orthotopic sites in SCID-bg mice and imaged over time. The LNCaP-luc-M6 cells formed subcutaneous and orthotopic tumors in SCID-bg mice, and nearly all tumor-bearing animals developed pulmonary metastases. Early detection and temporal growth of primary tumors and metastatic lesions was successfully monitored by BLI. The LNCaP-luc-M6 cell line is a bioluminescent, hormone-sensitive prostate cancer cell line applicable for BLI studies to non-invasively monitor subcutaneous and orthotopic prostate tumor growth and metastasis in vivo. Copyright 2004 Wiley-Liss, Inc.

Profiling the immunotoxicity of chemicals based on in vitro evaluation by a combination of the Multi-ImmunoTox assay and the IL-8 Luc assay.

PubMed

Kimura, Yutaka; Fujimura, Chizu; Ito, Yumiko; Takahashi, Toshiya; Terui, Hitoshi; Aiba, Setsuya

2018-06-01

We established a luciferase reporter assay system, the Multi-ImmunoTox Assay (MITA), which can evaluate the effects of chemicals on the promoter activities of four cytokines: IL-2, IFN-γ, IL-1β, and IL-8. We previously reported that MITA correctly reflected the change in mRNA of human whole-blood cells treated with dexamethasone, cyclosporine, FK506, or several other immunosuppressive drugs. In this study, we combined MITA with the IL-8 Luc assay to detect skin sensitization chemicals (OECD 442E) (modified MITA: mMITA) and established a data set of 60 chemicals examined by mMITA. Using the mMITA results, chemicals can be classified based on the lowest observed effect level (LOEL) of chemicals in suppressing or augmenting the promoter activities of the four cytokines. Moreover, we demonstrated that K-means clustering and hierarchical clustering of the 60 chemicals based on the LOEL for their effects on IL-2 and IL-8 promoter activities and the judgment by the IL-8 Luc assay resulted in the same 6-cluster solution: cluster 1 with preferential suppression of IL-8, cluster 2 with suppression of IL-2 and a positive IL-8 Luc assay result, cluster 3 with suppression of both IL-2 and IL-8, cluster 4 with no effects on IL-2 or IL-8 and a negative IL-8 Luc assay result, cluster 5 with suppression of both IL-2 and IL-8 and a negative IL-8 Luc assay result, and cluster 6 with preferential suppression of IL-8. These data suggest that mMITA is a promising novel high-throughput approach for detecting unrecognized immunological effects of chemicals and for profiling their immunotoxic effects.

Prediction of in vivo developmental toxicity by combination of Hand1-Luc embryonic stem cell test and metabolic stability test with clarification of metabolically inapplicable candidates.

PubMed

Nagahori, Hirohisa; Suzuki, Noriyuki; Le Coz, Florian; Omori, Takashi; Saito, Koichi

2016-09-30

Hand1-Luc Embryonic Stem Cell Test (Hand1-Luc EST) is a promising alternative method for evaluation of developmental toxicity. However, the problems of predictivity have remained due to appropriateness of the solubility, metabolic system, and prediction model. Therefore, we assessed the usefulness of rat liver S9 metabolic stability test using LC-MS/MS to develop new prediction model. A total of 71 chemicals were analyzed by measuring cytotoxicity and differentiation toxicity, and highly reproducible (CV=20%) results were obtained. The first prediction model was developed by discriminant analysis performed on a full dataset using Hand1-Luc EST, and 66.2% of the chemicals were correctly classified by the cross-validated classification. A second model was developed with additional descriptors obtained from the metabolic stability test to calculate hepatic availability, and an accuracy of 83.3% was obtained with applicability domain of 50.7% (=36/71) after exclusion of 22 metabolically inapplicable candidates, which potentially have a metabolic activation property. A step-wise prediction scheme with combination of Hand1-Luc EST and metabolic stability test was therefore proposed. The current results provide a promising in vitro test method for accurately predicting in vivo developmental toxicity. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

An ultrasensitive NanoLuc-based luminescence system for monitoring Plasmodium berghei throughout its life cycle.

PubMed

De Niz, Mariana; Stanway, Rebecca R; Wacker, Rahel; Keller, Derya; Heussler, Volker T

2016-04-21

Bioluminescence imaging is widely used for cell-based assays and animal imaging studies, both in biomedical research and drug development. Its main advantages include its high-throughput applicability, affordability, high sensitivity, operational simplicity, and quantitative outputs. In malaria research, bioluminescence has been used for drug discovery in vivo and in vitro, exploring host-pathogen interactions, and studying multiple aspects of Plasmodium biology. While the number of fluorescent proteins available for imaging has undergone a great expansion over the last two decades, enabling simultaneous visualization of multiple molecular and cellular events, expansion of available luciferases has lagged behind. The most widely used bioluminescent probe in malaria research is the Photinus pyralis firefly luciferase, followed by the more recently introduced Click-beetle and Renilla luciferases. Ultra-sensitive imaging of Plasmodium at low parasite densities has not been previously achieved. With the purpose of overcoming these challenges, a Plasmodium berghei line expressing the novel ultra-bright luciferase enzyme NanoLuc, called PbNLuc has been generated, and is presented in this work. NanoLuc shows at least 150 times brighter signal than firefly luciferase in vitro, allowing single parasite detection in mosquito, liver, and sexual and asexual blood stages. As a proof-of-concept, the PbNLuc parasites were used to image parasite development in the mosquito, liver and blood stages of infection, and to specifically explore parasite liver stage egress, and pre-patency period in vivo. PbNLuc is a suitable parasite line for sensitive imaging of the entire Plasmodium life cycle. Its sensitivity makes it a promising line to be used as a reference for drug candidate testing, as well as the characterization of mutant parasites to explore the function of parasite proteins, host-parasite interactions, and the better understanding of Plasmodium biology. Since the substrate

Do convection-permitting models improve the representation of the impact of LUC?

NASA Astrophysics Data System (ADS)

Vanden Broucke, Sam; Van Lipzig, Nicole

2017-10-01

In this study we assess the added value of convection permitting scale (CPS) simulations in studies using regional climate models to quantify the bio-geophysical climate impact of land-use change (LUC). To accomplish this, a comprehensive model evaluation methodology is applied to both non-CPS and CPS simulations. The main characteristics of the evaluation methodology are (1) the use of paired eddy-covariance site observations (forest vs open land) and (2) a simultaneous evaluation of all surface energy budget components. Results show that although generally satisfactory, non-CPS simulations fall short of completely reproducing the observed LUC signal because of three key biases. CPS scale simulations succeed at significantly reducing two of these biases, namely, those in daytime shortwave radiation and daytime sensible heat flux. Also, CPS slightly reduces a third bias in nighttime incoming longwave radiation. The daytime improvements can be attributed partially to the switch from parameterized to explicit convection, the associated improvement in the simulation of afternoon convective clouds, and resulting surface energy budget and atmospheric feedbacks. Also responsible for the improvements during daytime is a better representation of surface heterogeneity and thus, surface roughness. Meanwhile, the modest nighttime longwave improvement can be attributed to increased vertical atmospheric resolution. However, the model still fails at reproducing the magnitude of the observed nighttime longwave difference. One possible explanation for this persistent bias is the nighttime radiative effect of biogenic volatile organic compound emissions over the forest site. A correlation between estimated emission rates and the observed nighttime longwave difference, as well as the persistence of the longwave bias provide support for this hypothesis. However, more research is needed to conclusively determine if the effect indeed exists.

Prediction of Protein-Protein Interactions by NanoLuc-Based Protein-Fragment Complementation Assay | Office of Cancer Genomics

Cancer.gov

The CTD2 Center at Emory has developed a new NanoLuc®-based protein-fragment complementation assay (NanoPCA) which allows the detection of novel protein-protein interactions (PPI). NanoPCA allows the study of PPI dynamics with reversible interactions.  Read the abstract. Experimental Approaches Read the detailed Experimetnal Approaches. 

Advancing bioluminescence imaging technology for the evaluation of anticancer agents in the MDA-MB-435-HAL-Luc mammary fat pad and subrenal capsule tumor models.

PubMed

Zhang, Cathy; Yan, Zhengming; Arango, Maria E; Painter, Cory L; Anderes, Kenna

2009-01-01

Tumors grafted s.c. or under the mammary fat pad (MFP) rarely develop efficient metastasis. By applying bioluminescence imaging (BLI) technology, the MDA-MB-435-HAL-Luc subrenal capsule (SRC) model was compared with the MFP model for disease progression, metastatic potential, and response to therapy. The luciferase-expressing MDA-MB-435-HAL-Luc cell line was used in both MFP and SRC models. BLI technology allowed longitudinal assessment of disease progression and the therapeutic response to PD-0332991, Avastin, and docetaxel. Immunohistochemical analysis of Ki67 and CD31 staining in the primary tumors was compared in these models. Caliper measurement was used in the MFP model to validate the BLI quantification of primary tumors. The primary tumors in MDA-MB-435-HAL-Luc MFP and SRC models displayed comparable growth rates and vascularity. However, tumor-bearing mice in the SRC model developed lung metastases much earlier (4 weeks) than in the MFP model (>7 weeks), and the metastatic progression contributed significantly to the survival time. In the MFP model, BLI and caliper measurements were comparable for quantifying palpable tumors, but BLI offered an advantage for detecting the primary tumors that fell below a palpable threshold and for visualizing metastases. In the SRC model, BLI allowed longitudinal assessment of the antitumor and antimetastatic effects of PD-0332991, Avastin, and docetaxel, and the results correlated with the survival benefits of these agents. The MDA-MB-435-HAL-Luc SRC model and the MFP model displayed differences in disease progression. BLI is an innovative approach for developing animal models and creates opportunities for improving preclinical evaluations of anticancer agents.

STANDARDIZATION AND VALIDATION OF ADENOVIRAL TRANSDUCTION OF AN ANDROGEN RECEPTOR POSITIVE CELL LINE WITH AN MMTV-LUC REPORTER FOR ENDOCRINE SCREENING

EPA Science Inventory

Standardization and Validation of Adenoviral Transduction of an Androgen Receptor Positive Cell Line with an MMTV-Luc Reporter for Endocrine Screening P. Hartig, K . Bobseine,
M. Cardon, C. Lambright and L. E. Gray, Jr. USEPA, Reproductive Toxicology Division, NHEERL, RTP, NC...

Effect of sodium-alginate and laminaran on Salmonella Typhimurium infection in human enterocyte-like HT-29-Luc cells and BALB/c mice.

PubMed

Kuda, Takashi; Kosaka, Misa; Hirano, Shino; Kawahara, Miho; Sato, Masahiro; Kaneshima, Tai; Nishizawa, Makoto; Takahashi, Hajime; Kimura, Bon

2015-07-10

Brown algal polysaccharides such as alginate, polymers of uronic acids, and laminaran, beta-1,3 and 1,6-glucan, can be fermented by human intestinal microbiota. To evaluate the effects of these polysaccharides on infections caused by food poisoning pathogens, we investigated the adhesion and invasion of pathogens (Salmonella Typhimurium, Listeria monocytogenes and Vibrio parahaemolyticus) in human enterocyte-like HT-29-Luc cells and in infections caused in BALB/c mice. Both sodium Na-alginate and laminaran (0.1% each) inhibited the adhesion of the pathogens to HT-29-Luc cells by approximately 70-90%. The invasion of S. Typhimurium was also inhibited by approximately 70 and 80% by Na-alginate and laminaran, respectively. We observed that incubation with Na-alginate for 18 h increased the transepithelial electrical resistance of HT-29-Luc monolayer cells. Four days after inoculation with 7 log CFU/mouse of S. Typhimurium, the faecal pathogen count in mice that were not fed polysaccharides (control mice) was about 6.5 log CFU/g while the count in mice that were fed Na-alginate had decreased to 5.0 log CFU/g. The liver pathogen count, which was 4.1 log CFU/g in the control mice, was also decreased in mice that were fed Na-alginate. In contrast, the mice that were fed laminaran exhibited a more severe infection than that exhibited by control mice. Copyright © 2015 Elsevier Ltd. All rights reserved.

Emory University: Prediction of Protein-Protein Interactions by NanoLuc-Based Protein-Fragment Complementation Assay | Office of Cancer Genomics

Cancer.gov

The CTD2 Center at Emory has developed a new NanoLuc®-based protein-fragment complementation assay (NanoPCA) which allows the detection of novel protein-protein interactions (PPI). NanoPCA allows the study of PPI dynamics with reversible interactions.  Read the abstract. Experimental Approaches Read the detailed Experimetnal Approaches. 

[Influence of MSA on cell growth and spontaneousn metastasis of L9981-Luc lung cancer transplanted model in nude mice by bioluminescence imaging].

PubMed

Ren, Yuanrong; Wang, Yuli; Liu, Hongyu; Yan, Huiqin; Chen, Jun; Hou, Mei; Li, Weimin; Fan, Yaguang; Zhou, Qinghua

2013-02-01

Methylseleninic acid (MSA) is an artificially developed selenium compound. It has been proven that MSA could inhibit growth and metastasis on many tumor cells. This study investigated whether MSA has an impact on the growth and metastasis of L9981-Luc lung cancer transplanted model in nude mice or not. A transplantated tumor model was established in nude mice. Fifteen nude mice were randomly divided into three groups: the control group treated with normal saline (0.2 mL/d), the MSA group treated with MSA solution (0.2 mL), and the cisplatin (DDP) group injected intraperitoneally with DDP (4 mg/kg/w). Inhibition of MSA on tumor growth and tumor metastasis was observed using the IVIS Imaging System 200 Series. A significant difference was obserced in the primary tumor bioluminescence among the three groups (P=0.002) on 21 days post-inoculation. Primary tumor bioluminescence in the DDP group (P=0.001) and in the MSA group (P=0.031) was significantly lower than that in the control group (P=0.001). No significant difference in the metastasis bioluminescence of the thoracic area was indicated among the three groups (P>0.05). MSA can inhibit the growth of planted tumor of transgenic lung cancer cell lines L9981-Luc in nude mice. MSA may also suppress the distant metastasis of the transplanted tumor of transgenic lung cancer cell lines L9981-Luc in nude mice.

NPY-Induced Phase Shifts of PER2::LUC Rhythms are Mediated by Long-Term Suppression of Neuronal Excitability in a Phase-Specific Manner

PubMed Central

Besing, Rachel C.; Hablitz, Lauren M.; Paul, Jodi R.; Johnson, Russell L.; Prosser, Rebecca A.; Gamble, Karen L.

2013-01-01

Endogenous circadian rhythms are entrained to the 24-h light/dark cycle by both light and nonphotic stimuli. During the day, nonphotic stimuli, such as novel-wheel induced exercise, produce large phase advances. Neuropeptide Y (NPY) release from the thalamus onto suprachiasmatic nucleus (SCN) neurons at least partially mediates this nonphotic signal. We examined the hypothesis that NPY-induced phase advances are accompanied by suppression of PER2 and are mediated by long-term depression of neuronal excitability in a phase-specific manner. First, we found that NPY-induced phase advances in PER2::LUC SCN cultures are largest when NPY (2.35 µM) is given in the early part of the day (circadian time [CT] 0–6). In addition, PER2::LUC levels in NPY-treated (compared to vehicle-treated) samples were suppressed beginning 6–7 h after treatment. Similar NPY application to organotypic Per1::GFP SCN cultures resulted in long-term suppression of spike rate of GFP+ cells when slices were treated with NPY during the early or middle of the day (zeitgeber time [ZT] 2 or 6), but not during the late day (ZT 10). Furthermore, 1-h bath application of NPY to acute SCN brain slices decreased general neuronal activity measured through extracellular recordings. Finally, NPY-induced phase advances of PER2::LUC rhythms were blocked by latent depolarization with 34.5 mM [K+] 3 h after NPY application. These results suggest that NPY-induced phase advances may be mediated by long-term depression of neuronal excitability. This model is consistent with findings in other brain regions that NPY-induced persistent hyperpolarization underlies mechanisms of energy homeostasis, anxiety-related behavior, and thalamocortical synchronous firing. PMID:22324550

Effect of Lactobacillus plantarum Tennozu-SU2 on Salmonella Typhimurium Infection in Human Enterocyte-Like HT-29-Luc Cells and BALB/c Mice.

PubMed

Hirano, Shino; Yokota, Yasushi; Eda, Mika; Kuda, Takashi; Shikano, Ayane; Takahashi, Hajime; Kimura, Bon

2017-03-01

The probiotic properties and inhibitory effect on Salmonella Typhimurium adhesion on human enterocyte-like HT-29-Luc cells of three Lactobacillus plantarum strains isolated from fermented fish, beach sand and a coastal plant were determined. Compared with the type strain L. plantarum NBRC 15891 T , which was isolated from pickled cabbage, L. plantarum Tennozu-SU2 isolated from the acorn of a coastal tree showed high autoaggregation in de Man, Rogosa and Sharpe (MRS) broth and an antagonistic effect against S. Typhimurium in brain heart infusion (BHI) broth. Furthermore, heat-killed L. plantarum Tennozu-SU2 cells inhibited S. Typhimurium adhesion on HT-29-Luc cells. Both live and heat-killed L. plantarum Tennozu-SU2 cells showed an inhibitory effect on gut colonisation in BALB/c mice, as assessed by viable Salmonella count in faecal samples and by invasion into liver and spleen tissues. The properties shown in this study suggest that L. plantarum Tennozu-SU2 is useful as a starter and probiotic bacteria in functional food material.

NanoLuc reporter for dual luciferase imaging in living animals.

PubMed

Stacer, Amanda C; Nyati, Shyam; Moudgil, Pranav; Iyengar, Rahul; Luker, Kathryn E; Rehemtulla, Alnawaz; Luker, Gary D

2013-10-01

Bioluminescence imaging is widely used for cell-based assays and animal imaging studies in biomedical research and drug development, capitalizing on the high signal to background of this technique. A relatively small number of luciferases are available for imaging studies, substantially limiting the ability to image multiple molecular and cellular events, as done commonly with fluorescence imaging. To advance dual reporter bioluminescence molecular imaging, we tested a recently developed, adenosine triphosphate–independent luciferase enzyme from Oplophorus gracilirostris (NanoLuc [NL]) as a reporter for animal imaging. We demonstrated that NL could be imaged in superficial and deep tissues in living mice, although the detection of NL in deep tissues was limited by emission of predominantly blue light by this enzyme. Changes in bioluminescence from NL over time could be used to quantify tumor growth, and secreted NL was detectable in small volumes of serum. We combined NL and firefly luciferase reporters to quantify two key steps in transforming growth factor β signaling in intact cells and living mice, establishing a novel dual luciferase imaging strategy for quantifying signal transduction and drug targeting. Our results establish NL as a new reporter for bioluminescence imaging studies in intact cells and living mice that will expand imaging of signal transduction in normal physiology, disease, and drug development.

NanoLuc Reporter for Dual Luciferase Imaging in Living Animals

PubMed Central

Stacer, Amanda C.; Nyati, Shyam; Moudgil, Pranav; Iyengar, Rahul; Luker, Kathryn E.; Rehemtulla, Alnawaz; Luker, Gary D.

2014-01-01

Bioluminescence imaging is utilized widely for cell-based assays and animal imaging studies in biomedical research and drug development, capitalizing on high signal-to-background of this technique. A relatively small number of luciferases are available for imaging studies, substantially limiting the ability to image multiple molecular and cellular events as done commonly with fluorescence imaging. To advance dual reporter bioluminescence molecular imaging, we tested a recently developed, ATP-independent luciferase enzyme from Oplophorus gracilirostris (NanoLuc, NL) as a reporter for animal imaging. We demonstrated that NL could be imaged in superficial and deep tissues in living mice, although detection of NL in deep tissues was limited by emission of predominantly blue light by this enzyme. Changes in bioluminescence from NL over time could be used to quantify tumor growth, and secreted NL was detectable in small volumes of serum. We combined NL and firefly luciferase reporters to quantify two key steps in TGF-β signaling in intact cells and living mice, establishing a novel dual luciferase imaging strategy for quantifying signal transduction and drug targeting. Our results establish NL as new reporter for bioluminescence imaging studies in intact cells and living mice that will expand imaging of signal transduction in normal physiology, disease, and drug development. PMID:24371848

Simultaneous imaging of temporal changes of NF-κB activity and viable tumor cells in Huh7/NF-κB-tk-luc2/rfp tumor-bearing mice.

PubMed

Wang, Wei-Hsun; Chiang, I-Tsang; Liu, Yu-Chang; Hsu, Fei-Ting; Chen, Hong-Wen; Chen, Chuan-Lin; Lee, Yi-Jang; Lin, Wuu-Jyh; Hwang, Jeng-Jong

2013-01-01

Few studies have reported that the effect of sorafenib on advanced human hepatocellular carcinoma (HCC) is taking place via the inhibition of NF-κB signal transduction. Here we constructed a human HCC Huh7 stable clone with NF-κB-responsive element to drive dual reporter genes, herpes simplex virus thymidine kinase (tk) and firefly luciferase (luc2), and co-transfected with a third red fluorescent protein (rfp) gene, renamed as Huh7/NF-κB-tk-luc2/rfp cells, and combined with bioluminescent imaging (BLI) and red fluorescent protein imaging (RFPI) to monitor the effect of sorafenib on NF-κB activation and tumor inhibition. The results show that sorafenib could suppress the NF-κB-DNA binding activity, and the expression of downstream effector proteins. Notably, the relative photon fluxes obtained from RFPI and BLI, which represent the viable tumor cells and cells with NF-κB activation, decreased after sorafenib treatment by 50 to 65%, and 87.5 to >90%, respectively, suggesting that NF-κB activation is suppressed in viable HCC cells by sorafenib. Simultaneous molecular imaging of the temporal change of NF-κB activity and of viable cells in the same Huh7/NF-κB-tk-luc2/rfp tumors of the animal may reflect the real status of NF-κB activity and the viable tumor cells at the time of imaging.

Effects of a non thermal plasma treatment alone or in combination with gemcitabine in a MIA PaCa2-luc orthotopic pancreatic carcinoma model.

PubMed

Brullé, Laura; Vandamme, Marc; Riès, Delphine; Martel, Eric; Robert, Eric; Lerondel, Stéphanie; Trichet, Valérie; Richard, Serge; Pouvesle, Jean-Michel; Le Pape, Alain

2012-01-01

Pancreatic tumors are the gastrointestinal cancer with the worst prognosis in humans and with a survival rate of 5% at 5 years. Nowadays, no chemotherapy has demonstrated efficacy in terms of survival for this cancer. Previous study focused on the development of a new therapy by non thermal plasma showed significant effects on tumor growth for colorectal carcinoma and glioblastoma. To allow targeted treatment, a fibered plasma (Plasma Gun) was developed and its evaluation was performed on an orthotopic mouse model of human pancreatic carcinoma using a MIA PaCa2-luc bioluminescent cell line. The aim of this study was to characterize this pancreatic carcinoma model and to determine the effects of Plasma Gun alone or in combination with gemcitabine. During a 36 days period, quantitative BLI could be used to follow the tumor progression and we demonstrated that plasma gun induced an inhibition of MIA PaCa2-luc cells proliferation in vitro and in vivo and that this effect could be improved by association with gemcitabine possibly thanks to its radiosensitizing properties.

Effects of a Non Thermal Plasma Treatment Alone or in Combination with Gemcitabine in a MIA PaCa2-luc Orthotopic Pancreatic Carcinoma Model

PubMed Central

Brullé, Laura; Vandamme, Marc; Riès, Delphine; Martel, Eric; Robert, Eric; Lerondel, Stéphanie; Trichet, Valérie; Richard, Serge; Pouvesle, Jean-Michel; Le Pape, Alain

2012-01-01

Pancreatic tumors are the gastrointestinal cancer with the worst prognosis in humans and with a survival rate of 5% at 5 years. Nowadays, no chemotherapy has demonstrated efficacy in terms of survival for this cancer. Previous study focused on the development of a new therapy by non thermal plasma showed significant effects on tumor growth for colorectal carcinoma and glioblastoma. To allow targeted treatment, a fibered plasma (Plasma Gun) was developed and its evaluation was performed on an orthotopic mouse model of human pancreatic carcinoma using a MIA PaCa2-luc bioluminescent cell line. The aim of this study was to characterize this pancreatic carcinoma model and to determine the effects of Plasma Gun alone or in combination with gemcitabine. During a 36 days period, quantitative BLI could be used to follow the tumor progression and we demonstrated that plasma gun induced an inhibition of MIA PaCa2-luc cells proliferation in vitro and in vivo and that this effect could be improved by association with gemcitabine possibly thanks to its radiosensitizing properties. PMID:23300736

AKT1, LKB1, and YAP1 revealed as MYC interactors with NanoLuc-based protein-fragment complementation assay. | Office of Cancer Genomics

Cancer.gov

The c-Myc (MYC) transcription factor is a major cancer driver and a well-validated therapeutic target. However, directly targeting MYC has been challenging. Thus, identifying proteins that interact with and regulate MYC may provide alternative strategies to inhibit its oncogenic activity. Here we report the development of a NanoLuc®-based protein-fragment complementation assay (NanoPCA) and mapping of the MYC protein interaction hub in live mammalian cells.

Postchallenge Administration of Brincidofovir Protects Healthy and Immune-Deficient Mice Reconstituted with Limited Numbers of T Cells from Lethal Challenge with IHD-J-Luc Vaccinia Virus

PubMed Central

McCullough, Kevin Tyler; Cruz, Stephanie; Thomas, Antonia; Diaz, Claudia G.; Keilholz, Laurie; Grossi, Irma M.; Trost, Lawrence C.; Golding, Hana

2015-01-01

ABSTRACT Protection from lethality by postchallenge administration of brincidofovir (BCV, CMX001) was studied in normal and immune-deficient (nude, nu/nu) BALB/c mice infected with vaccinia virus (VACV). Whole-body bioluminescence imaging was used to record total fluxes in the nasal cavity, lungs, spleen, and liver and to enumerate pox lesions on tails of mice infected via the intranasal route with 105 PFU of recombinant IHD-J-Luc VACV expressing luciferase. Areas under the flux curve (AUCs) were calculated for individual mice to assess viral loads. A three-dose regimen of 20 mg/kg BCV administered every 48 h starting either on day 1 or day 2 postchallenge protected 100% of mice. Initiating BCV treatment earlier was more efficient in reducing viral loads and in providing protection from pox lesion development. All BCV-treated mice that survived challenge were also protected from rechallenge with IHD-J-Luc or WRvFire VACV without additional treatment. In immune-deficient mice, BCV protected animals from lethality and reduced viral loads while animals were on the drug. Viral recrudescence occurred within 4 to 9 days, and mice succumbed ∼10 to 20 days after treatment termination. Nude mice reconstituted with 105 T cells prior to challenge with 104 PFU of IHD-J-Luc and treated with BCV postchallenge survived the infection, cleared the virus from all organs, and survived rechallenge with 105 PFU of IHD-J-Luc VACV without additional BCV treatment. Together, these data suggest that BCV protects immunocompetent and partially T cell-reconstituted immune-deficient mice from lethality, reduces viral dissemination in organs, prevents pox lesion development, and permits generation of VACV-specific memory. IMPORTANCE Mass vaccination is the primary element of the public health response to a smallpox outbreak. In addition to vaccination, however, antiviral drugs are required for individuals with uncertain exposure status to smallpox or for whom vaccination is contraindicated

Food-anticipatory activity and liver per1-luc activity in diabetic transgenic rats

NASA Technical Reports Server (NTRS)

Davidson, Alec J.; Stokkan, Karl-Arne; Yamazaki, Shin; Menaker, Michael

2002-01-01

The mammalian Per1 gene is an important component of the core cellular clock mechanism responsible for circadian rhythms. The rodent liver and other tissues rhythmically express Per1 in vitro but typically damp out within a few cycles. In the liver, the peak of this rhythm occurs in the late subjective night in an ad lib-fed rat, but will show a large phase advance in response to restricted availability of food during the day. The relationship between this shift in the liver clock and food-anticipatory activity (FAA), the circadian behavior entrained by daily feeding, is currently unknown. Insulin is released during feeding in mammals and could serve as an entraining signal to the liver. To test the role of insulin in the shift in liver Per1 expression and the generation of FAA, per-luciferase transgenic rats were made diabetic with a single injection of streptozotocine. Following 1 week of restricted feeding and locomotor activity monitoring, liver was collected for per-luc recording. In two separate experiments, FAA emerged and liver Per1 phase-shifted in response to daytime 8-h food restriction. The results rule out insulin as a necessary component of this system.

Electron-phonon interaction in the binary superconductor lutetium carbide LuC2 via first-principles calculations

NASA Astrophysics Data System (ADS)

Dilmi, S.; Saib, S.; Bouarissa, N.

2018-06-01

Structural, electronic, electron-phonon coupling and superconducting properties of the intermetallic compound LuC2 are investigated by means of ab initio pseudopotential plane wave method within the generalized gradient approximation. The calculated equilibrium lattice parameters yielded a very good accord with experiment. There is no imaginary phonon frequency in the whole Brillouin zone supporting thus the dynamical stability in the material of interest. The average electron-phonon coupling parameter is found to be 0.59 indicating thus a weak-coupling BCS superconductor. Using a reasonable value of μ* = 0.12 for the effective Coulomb repulsion parameter, the superconducting critical temperature Tc is found to be 3.324 which is in excellent agreement with the experimental value of 3.33 K. The effect of the spin-orbit coupling on the superconducting properties of the material of interest has been examined and found to be weak.

Choosing a College Major: Factors that Might Influence the Way Students Make Decisions

ERIC Educational Resources Information Center

Lee, Wei-Chun Vanessa

2009-01-01

This current study investigated Janis and Mann's (1977) Conflict Model of Decision Making. Specifically, Janis and Mann's model was tested to examine decision-making styles (coping patterns) and students who either have already decided or who have yet to decide on their college major. Furthermore, the current study is aimed to expand Janis and…

Exploiting NanoLuc luciferase for smartphone-based bioluminescence cell biosensor for (anti)-inflammatory activity and toxicity.

PubMed

Cevenini, Luca; Calabretta, Maria Maddalena; Lopreside, Antonia; Tarantino, Giuseppe; Tassoni, Annalisa; Ferri, Maura; Roda, Aldo; Michelini, Elisa

2016-12-01

The availability of smartphones with high-performance digital image sensors and processing power has completely reshaped the landscape of point-of-need analysis. Thanks to the high maturity level of reporter gene technology and the availability of several bioluminescent proteins with improved features, we were able to develop a bioluminescence smartphone-based biosensing platform exploiting the highly sensitive NanoLuc luciferase as reporter. A 3D-printed smartphone-integrated cell biosensor based on genetically engineered Hek293T cells was developed. Quantitative assessment of (anti)-inflammatory activity and toxicity of liquid samples was performed with a simple and rapid add-and-measure procedure. White grape pomace extracts, known to contain several bioactive compounds, were analyzed, confirming the suitability of the smartphone biosensing platform for analysis of untreated complex biological matrices. Such approach could meet the needs of small medium enterprises lacking fully equipped laboratories for first-level safety tests and rapid screening of new bioactive products. Graphical abstract Smartphone-based bioluminescence cell biosensor.

Illness as the saturated phenomenon: the contribution of Jean-Luc Marion.

PubMed

Grīnfelde, Māra

2018-05-24

During the last few decades, many thinkers have advocated for the importance of the phenomenological approach in developing the understanding of the lived experience of illness. In their attempts, they have referred to ideas found in the history of phenomenology, most notably, in the works of Edmund Husserl, Martin Heidegger, Maurice Merleau-Ponty and Jean-Paul Sartre. The aim of this paper is to sketch out an interpretation of illness based on a yet unexplored conceptual framework of the phenomenology of French thinker Jean-Luc Marion. Focusing on concepts of the saturated phenomenon and flesh, the paper develops an interpretation of illness as the saturated phenomenon, which highlights a variety of dimensions of illness already elaborated within the phenomenology of medicine, such as the affective dimension of illness, the disruptive dimension of illness, the transformed perception of the self in illness, mineness of flesh in illness and the inexpressible and hermeneutical dimension of illness. In addition to that, the paper explores some of the consequences the proposed interpretation of illness offers regarding the nature of illness and health. It is argued that illness in its essence is very similar to the experience of other saturated phenomena, suggesting that the difference between them does not lie within the character of the affective givenness, but rather within the dynamic relationship between the affective givenness and its conceptualization. It is also shown that the experience of health is compatible with the experience of saturation and thus is not limited to the tacit and harmonious background state.

NanoLuc: A Small Luciferase is Brightening up the Field of Bioluminescence

PubMed Central

Cai, Weibo

2016-01-01

The biomedical field has greatly benefited from the discovery of bioluminescent proteins. Currently, scientists employ bioluminescent systems for numerous biomedical applications, ranging from highly sensitive cellular assays to bioluminescence-based molecular imaging. Traditionally, these systems are based on Firefly and Renilla luciferases; however, the applicability of these enzymes is limited by their size, stability, and luminescence efficiency. NanoLuc (NLuc), a novel bioluminescence platform, offers several advantages over established systems, including enhanced stability, smaller size, and >150-fold increase in luminescence. In addition, the substrate for NLuc displays enhanced stability and lower background activity, opening up new possibilities in the field of bioluminescence imaging. The NLuc system is incredibly versatile and may be utilized for a wide array of applications. The increased sensitivity, high stability, and small size of the NLuc system have the potential to drastically change the field of reporter assays in the future. However, as with all such technology, NLuc has limitations (including a non-ideal emission for in vivo applications and its unique substrate) which may cause it to find restricted use in certain areas of molecular biology. As this unique technology continues to broaden, NLuc may have a significant impact in both preclinical and clinical fields, with potential roles in disease detection, molecular imaging, and therapeutic monitoring. This review will present the NLuc technology to the scientific community in a non-biased manner, allowing the audience to adopt their own views of this novel system. PMID:27045664

C3-Luc Cells Are an Excellent Model for Evaluation of Cellular Immunity following HPV16L1 Vaccination.

PubMed

Li, Li-Li; Wang, He-Rong; Zhou, Zhi-Yi; Luo, Jing; Wang, Xiao-Li; Xiao, Xiang-Qian; Zhou, Yu-Bai; Zeng, Yi

2016-01-01

C3 and TC-1 are the two model cell lines most commonly used in studies of vaccines and drugs against human papillomavirus (HPV) infection. Because C3 cells contain both the HPV16 E and L genes, but TC-1 cells contain only the HPV16 E genes, C3 cells are usually used as the model cell line in studies targeting the HPV16 L protein. However, expression of the L1 protein is difficult to detect in C3 cells using common methods. In our study, Short tandem repeat analysis (STR) was used to demonstrate that C3 cells are indeed derived from mice, PCR results show that HPV16 L1, E6 and E7 genes were detected in C3 genomic DNA, and RT-PCR results demonstrated that L1 transcription had occurred in C3 cells. However, the expression of C3 protein was not found in the results of western blot and immunohistochemistry (IHC). Growth and proliferation of C3 were inhibited by mice spleen lymphocytes that had been immunized with a vaccine against HPV16L1. The luciferase gene was integrated into C3 cells, and it was confirmed that addition of the exogenous gene had no effect on C3 cells by comparing cell growth and tumor formation with untransformed cells. Cells stably expressing luciferase (C3-luc) were screened and subcutaneously injected into the mice. Tumors became established and were observed using a Spectrum Pre-clinical in Vivo Imaging System. Tumor size of mice in the different groups at various time points was calculated by counting photons. The sensitivity of the animals to the vaccine was quantified by statistical comparison. Ten or 30 days following injection of the C3-luc cells, tumor size differed significantly between the PBS and vaccine groups, indicating that C3 cells were susceptible to vaccination even after tumors were formed in vivo.

Dietary exposure to polychlorinated dibenzo-p-dioxins and dibenzofurans via fish consumption and dioxin-like activity in fish determined by H4IIE-luc bioassay.

PubMed

Chan, Janet Kit Yan; Man, Yu Bon; Xing, Guan Hua; Wu, Sheng Chun; Murphy, Margaret B; Xu, Ying; Wong, Ming H

2013-10-01

Dietary exposure to polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs) via fish consumption in two major electronic (e) waste sites: Guiyu (GY), Guangdong Province and Taizhou (TZ), Zhejiang Province, and dioxin-like activity in fish determined by H4IIE-luc bioassay. In the present study, all fish were below EU's maximum allowable concentration in muscle of fish (4 pg WHO-TEQ/g wet wt), except crucian (4.28 pg WHO-TEQ/g wet wt) and silver carps (7.49 pg WHO-TEQ/g wet wt) collected from GY rivers. Moreover, the residual concentration in bighead carp collected from GY (2.15 pg WHO-TEQ/g wet wt) was close to the EU's action level (3 pg WHO-TEQ/g wet wt) which gives "early warning" to the competent authorities and operators to take measures to eliminate contamination. In addition, results indicated that the maximum human intake of PCDD/Fs via freshwater fish consumption in GY was 4.31 pg WHO-TEQ/kg bw/day, which exceeds the higher end of the tolerable daily intake recommended by the WHO, EC-SCF and JECFA (1-4, 2 and 2.3 pg WHO-TEQ/kg bw/day respectively). Furthermore, H4IIE-luc cell bioassay provides a very sensitive and cost-efficient screening tool for assessing the overall dioxin-like toxicity in the study, and is therefore valuable for high-throughput environmental monitoring studies. Copyright © 2012 Elsevier B.V. All rights reserved.

Forced rather than voluntary exercise entrains peripheral clocks via a corticosterone/noradrenaline increase in PER2::LUC mice

PubMed Central

Sasaki, Hiroyuki; Hattori, Yuta; Ikeda, Yuko; Kamagata, Mayo; Iwami, Shiho; Yasuda, Shinnosuke; Tahara, Yu; Shibata, Shigenobu

2016-01-01

Exercise during the inactive period can entrain locomotor activity and peripheral circadian clock rhythm in mice; however, mechanisms underlying this entrainment are yet to be elucidated. Here, we showed that the bioluminescence rhythm of peripheral clocks in PER2::LUC mice was strongly entrained by forced treadmill and forced wheel-running exercise rather than by voluntary wheel-running exercise at middle time during the inactivity period. Exercise-induced entrainment was accompanied by increased levels of serum corticosterone and norepinephrine in peripheral tissues, similar to the physical stress-induced response. Adrenalectomy with norepinephrine receptor blockers completely blocked the treadmill exercise-induced entrainment. The entrainment of the peripheral clock by exercise is independent of the suprachiasmatic nucleus clock, the main oscillator in mammals. The present results suggest that the response of forced exercise, but not voluntary exercise, may be similar to that of stress, and possesses the entrainment ability of peripheral clocks through the activation of the adrenal gland and the sympathetic nervous system. PMID:27271267

The impacts of different long-term fertilization regimes on the bioavailability of arsenic in soil: integrating chemical approach with Escherichia coli arsRp::luc-based biosensor.

PubMed

Hou, Qi-Hui; Ma, An-Zhou; Lv, Di; Bai, Zhi-Hui; Zhuang, Xu-Liang; Zhuang, Guo-Qiang

2014-07-01

An Escherichia coli arsRp::luc-based biosensor was constructed to measure the bioavailability of arsenic (As) in soil. In previous induction experiments, it produced a linear response (R (2) = 0.96, P < 0.01) to As from 0.05 to 5 μmol/L after a 2-h incubation. Then, both chemical sequential extraction, Community Bureau of Reference recommended sequential extraction procedures (BCR-SEPs) and E. coli biosensor, were employed to assess the impact of different long-term fertilization regimes containing N, NP, NPK, M (manure), and NPK + M treatments on the bioavailability of arsenic (As) in soil. Per the BCR-SEPs analysis, the application of M and M + NPK led to a significant (P < 0.01) increase of exchangeable As (2-7 times and 2-5 times, respectively) and reducible As (1.5-2.5 times and 1.5-2.3 times, respectively) compared with the no fertilization treated soil (CK). In addition, direct contact assay of E. coli biosensor with soil particles also supported that bioavailable As in manure-fertilized (M and M + NPK) soil was significantly higher (P < 0.01) than that in CK soil (7 and 9 times, respectively). Organic carbon may be the major factor governing the increase of bioavailable As. More significantly, E. coli biosensor-determined As was only 18.46-85.17 % of exchangeable As and 20.68-90.1 % of reducible As based on BCR-SEPs. In conclusion, NKP fertilization was recommended as a more suitable regime in As-polluted soil especially with high As concentration, and this E. coli arsRp::luc-based biosensor was a more realistic approach in assessing the bioavailability of As in soil since it would not overrate the risk of As to the environment.

Luciferase NanoLuc as a reporter for gene expression and protein levels in Saccharomyces cerevisiae.

PubMed

Masser, Anna E; Kandasamy, Ganapathi; Kaimal, Jayasankar Mohanakrishnan; Andréasson, Claes

2016-05-01

Reporter proteins are essential tools in the study of biological processes and are employed to monitor changes in gene expression and protein levels. Luciferases are reporter proteins that enable rapid and highly sensitive detection with an outstanding dynamic range. Here we evaluated the usefulness of the 19 kDa luciferase NanoLuc (Nluc), derived from the deep sea shrimp Oplophorus gracilirostris, as a reporter protein in yeast. Cassettes with codon-optimized genes expressing yeast Nluc (yNluc) or its destabilized derivative yNlucPEST have been assembled in the context of the dominant drug resistance marker kanMX. The reporter proteins do not impair the growth of yeast cells and exhibit half-lives of 40 and 5 min, respectively. The commercial substrate Nano-Glo® is compatible with detection of yNluc bioluminescence in < 50 cells. Using the unstable yNlucPEST to report on the rapid and transient expression of a heat-shock promoter (PCYC1-HSE ), we found a close match between the intensity of the bioluminescent signal and mRNA levels during both induction and decay. We demonstrated that the bioluminescence of yNluc fused to the C-terminus of a temperature-sensitive protein reports on its protein levels. In conclusion, yNluc and yNlucPEST are valuable new reporter proteins suitable for experiments with yeast using standard commercial substrate. © 2016 The Authors. Yeast published by John Wiley & Sons Ltd. © 2016 The Authors. Yeast published by John Wiley & Sons Ltd.

Expert Decision-Making in Naturalistic Environments: A Summary of Research

DTIC Science & Technology

2005-03-01

a number of descriptive decision theories arose (Plous, 1993). One of these is the rational choice model of decision - making (Janis & Mann, 1977...possible association between time pressure and increased levels of emotion . To date, the role played by emotion in decision - making has not been given... rational choice model seems to describe some decision events and Janis and Mann (1977) have highlighted emotion as a potential influence on decision

Was the Group Dynamic Phenomenon Groupthink Present on Board the USS Greeneville (SSN-772) When She Collided with the Japanese Fishing Vessel Ehime Maru?

DTIC Science & Technology

2002-05-31

Janis, professor of psychology at Yale, first presented his theory of groupthink. Janis introduced the term “groupthink” to refer to “a mode of thinking...thesis explores the groupthink phenomenon and utilizes Janis’ theory to come to the conclusion that groupthink was indeed present and was one of many...form a group and just as many conflicting theories that attempt to answer them. Is the group a fair representation of the individual members’ attitudes

Plasmodium falciparum Transfected with Ultra Bright NanoLuc Luciferase Offers High Sensitivity Detection for the Screening of Growth and Cellular Trafficking Inhibitors

PubMed Central

Elsworth, Brendan; Charnaud, Sarah C.; Sanders, Paul R.; Crabb, Brendan S.; Gilson, Paul R.

2014-01-01

Drug discovery is a key part of malaria control and eradication strategies, and could benefit from sensitive and affordable assays to quantify parasite growth and to help identify the targets of potential anti-malarial compounds. Bioluminescence, achieved through expression of exogenous luciferases, is a powerful tool that has been applied in studies of several aspects of parasite biology and high throughput growth assays. We have expressed the new reporter NanoLuc (Nluc) luciferase in Plasmodium falciparum and showed it is at least 100 times brighter than the commonly used firefly luciferase. Nluc brightness was explored as a means to achieve a growth assay with higher sensitivity and lower cost. In addition we attempted to develop other screening assays that may help interrogate libraries of inhibitory compounds for their mechanism of action. To this end parasites were engineered to express Nluc in the cytoplasm, the parasitophorous vacuole that surrounds the intraerythrocytic parasite or exported to the red blood cell cytosol. As proof-of-concept, these parasites were used to develop functional screening assays for quantifying the effects of Brefeldin A, an inhibitor of protein secretion, and Furosemide, an inhibitor of new permeation pathways used by parasites to acquire plasma nutrients. PMID:25392998

Operation Plan I-45 ICEBERG

DTIC Science & Technology

1945-02-01

8217COmIIAND OF MAJOR- GENERAL del VALLE: R. O, BARE, Col, USMC, ·C of S. ANNEXES: ABLE - Distribution B1iKER - Intelligence CHARLIE - Naval Gunfire Support Plan...3. ABLE Annex BAKER to Opn Plan 1-45, 1st MarDiv-(Rein) INTELLIGENCE 00015 1st Mar Div (Rein). 1990-5-80 In The Field 475/355 . 1100, 1OFeb, 1945. AP...distributed. (c) JANIS No. 86, August, 1944; JANIS No. 86 (Change 1), October 1944. (d) Engineer Intelligence Information of OKINAWA SHIMA prepared by Eng

Biochemical characteristics and gene expression profiles of two paralogous luciferases from the Japanese firefly Pyrocoelia atripennis (Coleoptera, Lampyridae, Lampyrinae): insight into the evolution of firefly luciferase genes.

PubMed

Bessho-Uehara, Manabu; Konishi, Kaori; Oba, Yuichi

2017-08-09

Two paralogous genes of firefly luciferase, Luc1 and Luc2, have been isolated from the species in two subfamilies, Luciolinae and Photurinae, of the family Lampyridae. The gene expression profiles have previously been examined only in the species of Luciolinae. Here we isolated Luc1 and Luc2 genes from the Japanese firefly Pyrocoelia atripennis. This is the first report of the presence of both Luc1 and Luc2 genes in the species of the subfamily Lampyrinae and of the exon-intron structure of Luc2 in the family Lampyridae. The luminescence of both gene products peaked at 547 nm under neutral buffer conditions, and the spectrum of Luc1, but not Luc2, was red-shifted under acidic conditions, as observed for Luc2 in the Luciolinae species. The semi-quantitative reverse transcription-polymerase chain reaction suggested that Luc1 was expressed in lanterns of all the stages except eggs, while Luc2 was expressed in the non-lantern bodies of eggs, prepupae, pupae, and female adults. These expression profiles are consistent with those in the Luciolinae species. Considering the distant phylogenetic relationship between Lampyrinae and Luciolinae in Lampyridae, we propose that fireflies generally possess two different luciferase genes and the biochemical properties and gene expression profiles for each paralog are conserved among lampyrid species.

Organic anion transporter 1 (OAT1/SLC22A6) enhances bioluminescence based on d-luciferin-luciferase reaction in living cells by facilitating the intracellular accumulation of d-luciferin.

PubMed

Furuya, Takahito; Takehara, Issey; Shimura, Asuka; Kishimoto, Hisanao; Yasujima, Tomoya; Ohta, Kinya; Shirasaka, Yoshiyuki; Yuasa, Hiroaki; Inoue, Katsuhisa

2018-01-15

Bioluminescence (BL) imaging based on d-luciferin (d-luc)-luciferase reaction allows noninvasive and real-time monitoring of luciferase-expressing cells. Because BL intensity depends on photons generated through the d-luc-luciferase reaction, an approach to increase intracellular levels of d-luc could improve the detection sensitivity. In the present study, we showed that organic anion transporter 1 (OAT1) is useful, as a d-luc transporter, in boosting the BL intensity in luciferase-expressing cells. Functional screening of several transporters showed that the expression of OAT1 in HEK293 cells stably expressing Pyrearinus termitilluminans luciferase (HEK293/eLuc) markedly enhanced BL intensity in the presence of d-luc. When OAT1 was transiently expressed in HEK293 cells, intracellular accumulation of d-luc was higher than that in control cells, and the specific d-luc uptake mediated by OAT1 was saturable with a Michaelis constant (K m ) of 0.23 μM. The interaction between OAT1 and d-luc was verified using 6-carboxyfluorescein, a typical substrate of OAT1, which showed that d-luc inhibited the uptake of 6-carboxyfluorescein mediated by OAT1. BL intensity was concentration-dependent at steady states in HEK293/eLuc cells stably expressing OAT1, and followed Michaelis-Menten kinetics with an apparent K m of 0.36 μM. In addition, the enhanced BL was significantly inhibited by OAT1-specific inhibitors. Thus, OAT1-mediated transport of d-luc could be a rate-limiting step in the d-luc-luciferase reaction. Furthermore, we found that expressing OAT1 in HEK293/eLuc cells implanted subcutaneously in mice also significantly increased the BL after intraperitoneal injection of d-luc. Our findings suggest that because OAT1 is capable of transporting d-luc, it can also be used to improve visualization and monitoring of luciferase-expressing cells. Copyright © 2017 Elsevier Inc. All rights reserved.

Development of a rapid and quantitative method for the analysis of viral entry and release using a NanoLuc luciferase complementation assay.

PubMed

Sasaki, Michihito; Anindita, Paulina D; Phongphaew, Wallaya; Carr, Michael; Kobayashi, Shintaro; Orba, Yasuko; Sawa, Hirofumi

2018-01-02

Subviral particles (SVPs) self-assemble and are released from cells transfected with expression plasmids encoding flavivirus structural proteins. Flavivirus-like particles (VLPs), consisting of flavivirus structural proteins and a subgenomic replicon, can enter cells and cause single-round infections. Neither SVPs or VLPs possess complete viral RNA genomes, therefore are replication-incompetent systems; however, they retain the capacity to fuse and bud from target cells and follow the same maturation process as whole virions. SVPs and VLPs have been previously employed in studies analyzing entry and release steps of viral life cycles. In this study, we have developed quantitative methods for the detection of cellular entry and release of SVPs and VLPs by applying a luciferase complementation assay based on the high affinity interaction between the split NanoLuc luciferase protein, LgBiT and the small peptide, HiBiT. We introduced HiBiT into the structural protein of West Nile virus and generated SVPs and VLPs harboring HiBiT (SVP-HiBiT and VLP-HiBiT, respectively). As SVP-HiBiT emitted strong luminescence upon exposure to LgBiT and its substrate, the nascently budded SVP-HiBiT in the supernatant was readily quantified by luminometry. Similarly, the cellular entry of VLP-HiBiT generated luminescence when VLP-HiBiT was infected into LgBiT-expressing cells. These methods utilizing SVP-HiBiT and VLP-HiBiT will facilitate research into life cycles of flaviviruses, including WNV. Copyright © 2017 Elsevier B.V. All rights reserved.

Sexually immature male ERE-Luc reporter mice to assess low dose estrogen-like effects of CdCl2 versus dietary Cd

PubMed Central

Ramachandran, Balaji; Rizzi, Nicoletta; Maggi, Adriana

2014-01-01

CdCl2 salt is widely used in exposure oriented studies, while the biological exposure of Cadmium (Cd) occurs mostly through diet. Hence, we designed a in vivo imaging methodology with sexually immature male ERE-Luc reporter mice to test the estrogen-like (EL) effects of Cd as a natural component in wheat and flax bread based diets (containing 17.57 and 49.22 ug/kg Cd concentrations respectively) and CdCl2 per-oral dose of 1 ug/kg/bw/day. Total exposure of ingested and % bioaccumulation of Cd in selected organs were estimated as 547 ng (4.4%), 776 ng (0.3%) and 2131.8 ng (0.1%) corresponding to CdCl2, wheat and flax bread based diet treatments respectively. Cd from CdCl2 bioaccumulated more readily, despite the exposure of Cd is higher with bread based diets. Longitudinal in vivo imaging did not reveal significant changes in luciferase activity. White adipose tissue (WAT) and prostate were identified as novel target organs of Cd. Indeed, the rest of the observed EL effects, endogenous target gene expression and necropsy findings are not consistent to any particular organ or treatment. This implies that, the observed EL effects due to low doses of Cd (either from CdCl2 or dietary form) occur only as subtle changes at the molecular level, but inadequate to cause significant changes at the anatomo-pathological level during the 21 day exposure period. The study demonstrates the sensitivity of the methodology to assess EL effects of food embedded Cd and underlines the limitations of directly extrapolating the results of suspected chemicals in their pure form to dietary exposure scenarios. PMID:24795841

Land-use change and greenhouse gas emissions from corn and cellulosic ethanol

PubMed Central

2013-01-01

Background The greenhouse gas (GHG) emissions that may accompany land-use change (LUC) from increased biofuel feedstock production are a source of debate in the discussion of drawbacks and advantages of biofuels. Estimates of LUC GHG emissions focus mainly on corn ethanol and vary widely. Increasing the understanding of LUC GHG impacts associated with both corn and cellulosic ethanol will inform the on-going debate concerning their magnitudes and sources of variability. Results In our study, we estimate LUC GHG emissions for ethanol from four feedstocks: corn, corn stover, switchgrass, and miscanthus. We use new computable general equilibrium (CGE) results for worldwide LUC. U.S. domestic carbon emission factors are from state-level modelling with a surrogate CENTURY model and U.S. Forest Service data. This paper investigates the effect of several key domestic lands carbon content modelling parameters on LUC GHG emissions. International carbon emission factors are from the Woods Hole Research Center. LUC GHG emissions are calculated from these LUCs and carbon content data with Argonne National Laboratory’s Carbon Calculator for Land Use Change from Biofuels Production (CCLUB) model. Our results indicate that miscanthus and corn ethanol have the lowest (−10 g CO2e/MJ) and highest (7.6 g CO2e/MJ) LUC GHG emissions under base case modelling assumptions. The results for corn ethanol are lower than corresponding results from previous studies. Switchgrass ethanol base case results (2.8 g CO2e/MJ) were the most influenced by assumptions regarding converted forestlands and the fate of carbon in harvested wood products. They are greater than miscanthus LUC GHG emissions because switchgrass is a lower-yielding crop. Finally, LUC GHG emissions for corn stover are essentially negligible and insensitive to changes in model assumptions. Conclusions This research provides new insight into the influence of key carbon content modelling variables on LUC GHG emissions

Development of a rapid immunoassay system: Luminescent detection of antigen-associated antibody-luciferase in the presence of a dye that absorbs light from free antibody-luciferase.

PubMed

Mori, Akihiro; Ojima-Kato, Teruyo; Fuchi, Shingo; Kaiya, Shinichi; Kojima, Takaaki; Nakano, Hideo

2017-12-01

In this report, we developed a rapid immunoassay system, designated the bioluminescent interference gathering optical (BINGO) assay, which required no time-consuming washing steps for removal of unbound antibodies. This system employed a luciferase (Luc)-conjugated antibody (LucAb) and a dye that absorbed light from the LucAb. The antigen-associated LucAb was localized by transfer of an antigen to the detector-side of a chamber where a detector photomultiplier tube (PMT) was installed. In contrast, the free LucAb was distributed throughout the solution, and the light emitted by the free LucAb was absorbed by the dye. Therefore, only light from LucAb associated with antigen could be detected by the PMT. The new system could be used to rapidly detect the amount of antigen-antibody-Luc complex by collecting steps, such as centrifugation or magnetic collection of antibody-coated magnetic beads. Proof-of-principle experiments were performed using a model system with streptavidin beads and biotinylated Luc. The feasibility of the system was demonstrated using magnetic beads coated with anti-Escherichia coli O157 antibody, enabling detection of 4 × 10 3  cells in only 15 min. Thus, this system may have applications in a variety of biomedical fields. Copyright © 2017 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

Chemical structure determination of DNA bases modified by active metabolites of lucidin-3-O-primeveroside.

PubMed

Ishii, Yuji; Okamura, Toshiya; Inoue, Tomoki; Fukuhara, Kiyoshi; Umemura, Takashi; Nishikawa, Akiyoshi

2010-01-01

Lucidin-3-O- primeveroside (LuP) is one of the components of madder root (Rubia tinctorum L.; MR) which is reported to be carcinogenic in the kidney and liver of rats. Since metabolism of LuP generates genotoxic compounds such as lucidin (Luc) and rubiadin (Rub), it is likely that LuP plays a key role in MR carcinogenesis. In the present study, the chemical structures of Luc-specific 2'-deoxyguanosine (dG) and 2'-deoxyadenosine (dA) adducts following the reactions of dG and dA with a Luc carbocation or quinone methide intermediate derived from Acetoxy-Luc were determined by liquid chromatography with photodiode array and electron spray ionizaion-mass spectrometry (LC-PDA-ESI/MS). The identification of the two measurable adducts as Luc-N(2)-dG and Luc-N(6)-dA was confirmed by NMR analysis. Subsequently, using a newly developed quantitative analytical method using LC-ESI/MS, the formation of Luc-N(2)-dG and Luc-N(6)-dA from the reaction of calf thymus DNA with Luc in the presence of S9 mixture was observed. The fact that this reaction with Rub also gave rise to the same dG and dA adducts strongly suggests that Rub genotoxicity involves a metabolic conversion to Luc. The precise determination of the modified DNA bases generated by LuP and the method for their analysis may contribute to further comprehension of the mode of action underlying carcinogenesis by MR and related anthraquinones.

[Establishment of a human bladder cancer cell line stably co-expressing hSPRY2 and luciferase genes and its subcutaneous tumor xenograft model in nude mice].

PubMed

Yin, Xiaotao; Li, Fanglong; Jin, Yipeng; Yin, Zhaoyang; Qi, Siyong; Wu, Shuai; Wang, Zicheng; Wang, Lin; Yu, Jiyun; Gao, Jiangping

2017-03-01

Objective To establish a human bladder cancer cell line stably co-expressing human sprouty2 (hSPRY2) and luciferase (Luc) genes simultaneously, and develop its subcutaneous tumor xenograft model in nude mice. Methods The hSPRY2 and Luc gene segments were amplified by PCR, and were cloned into lentiviral vector pCDH and pLVX respectively to produce corresponding lentivirus particles. The J82 human bladder cancer cells were infected with these two kinds of lentivirus particles, and then further screened by puromycin and G418. The expressions of hSPRY2 and Luc genes were detected by bioluminescence, immunofluorescence and Western blot analysis. The screened J82-hSPRY2/Luc cells were injected subcutaneously into BALB/c nude mice, and the growth of tumor was monitored dynamically using in vivo fluorescence imaging system. Results J82-hSPRY2/Luc cell line stably expressing hSPRY2 and Luc genes was established successfully. Bioluminescence, immunofluorescence and Western blot analysis validated the expressions of hSPRY2 and Luc genes. The in vivo fluorescence imaging system showed obvious fluorescence in subcutaneous tumor xenograft in nude mice. Conclusion The J82-hSPRY2/Luc bladder cancer cell line and its subcutaneous tumor xenograft model in nude mice have been established successfully.

The Impact of Pre-Industrial Land Use Change on Atmospheric Composition and Aerosol Radiative Forcing.

NASA Astrophysics Data System (ADS)

Hamilton, D. S.; Carslaw, K. S.; Spracklen, D. V.; Folberth, G.; Kaplan, J. O.; Pringle, K.; Scott, C.

2015-12-01

Anthropogenic land use change (LUC) has had a major impact on the climate by altering the amount of carbon stored in vegetation, changing surface albedo and modifying the levels of both biogenic and pyrogenic emissions. While previous studies of LUC have largely focused on the first two components, there has recently been a recognition that changes to aerosol and related pre-cursor gas emissions from LUC are equally important. Furthermore, it has also recently been recognised that the pre-industrial (PI) to present day (PD) radiative forcing (RF) of climate from aerosol cloud interactions (ACI) due to anthropogenic emissions is highly sensitive to the amount of natural aerosol that was present in the PI. This suggests that anthropogenic RF from ACI may be highly sensitive to land-use in the PI. There are currently two commonly used baseline reference years for the PI; 1750 and 1860. Rapid LUC occurred between 1750 and 1860, with large reductions in natural vegetation cover in Eastern Northern America, Europe, Central Russia, India and Eastern China as well as lower reductions in parts of Brazil and Africa. This LUC will have led to significant changes in biogenic and fire emissions with implications for natural aerosol concentrations and PI-to-PD RF. The focus of this study is therefore to quantify the impact of LUC between 1750 and 1860 on aerosol concentrations and PI-to-PD RF calculations from ACI. We use the UK Met Office HadGEM3-UKCA coupled-chemistry-climate model to calculate the impacts of anthropogenic emissions and anthropogenic LUC on aerosol size distributions in both 1750 and 1860. We prescribe LUC using the KK10 historical dataset of land cover change. Monoterpene emissions are coupled directly to the prescribed LUC through the JULES land surface scheme in HadGEM3. Fire emissions from LUC were calculated offline using the fire module LPJ-LMfire in the Lund-Potsdam-Jena dynamic global vegetation model. To separate out the impacts of LUC from

Future land-use change emissions: CO2, BVOC and wildfire

NASA Astrophysics Data System (ADS)

Arneth, A.; Knorr, W.; Hantson, S.; Anthoni, P.; Szogs, S.

2015-12-01

Historical land-use (LUC) change is known to have been a large source of CO2 emissions, mostly from deforestation: the equivalent of around 1/3 of today's CO2 in the atmosphere arises from LUC. And LUC will continue into the future, although the expected area change, the type of LUC (deforestation vs. afforestation/reforestation) and regions where the LUC will take place will differ greatly, depending on the future scenario. But LUC is not only of importance for projecting emissions of CO2. It also affects greatly emissions of biogenic volatile organic carbon, and from wildfires - all of which are important for the quantification of precursor substances relevant to air quality, and interactions with climate change. We show here that accounting for future socio-economic developments and LUC scenarios has the potential to override climate change and effects of CO2 fertilisation on fire and BVOC emission, regionally and in some cases also globally. Simulation experiments with the dynamic global vegetation model LPJ-GUESS will be performed, covering the 20th and 21st century, and assessing a rage of future population growth, LUC and climate change scenarios. For wildfire emissions, we find that burned area and emissions depend greatly on the type of population growth scenario, and on the distribution of urban vs rural population. BVOC emissions depend greatly on the amount and location of deforestation vs the region and magnitude of forest expansion in response to warming, such as through expansion of vegetation in the northern hemisphere, and via reforestation/afforestation. LUC so far has not been given sufficient attention for simulations of future air quality-climate interactions. In terms of terrestrial precursor emissions of atmospherically reactive substances our simulations clearly demonstrate the importance of including LUC in combination with vegetation that responds dynamically to changes in climate and atmospheric CO2 levels.

Influence of land use on water quality in a tropical landscape: a multi-scale analysis

PubMed Central

Yackulic, Charles B.; Lim, Yili; Arce-Nazario, Javier A.

2015-01-01

There is a pressing need to understand the consequences of human activities, such as land transformations, on watershed ecosystem services. This is a challenging task because different indicators of water quality and yield are expected to vary in their responsiveness to large versus local-scale heterogeneity in land use and land cover (LUC). Here we rely on water quality data collected between 1977 and 2000 from dozens of gauge stations in Puerto Rico together with precipitation data and land cover maps to (1) quantify impacts of spatial heterogeneity in LUC on several water quality indicators; (2) determine the spatial scale at which this heterogeneity influences water quality; and (3) examine how antecedent precipitation modulates these impacts. Our models explained 30–58% of observed variance in water quality metrics. Temporal variation in antecedent precipitation and changes in LUC between measurements periods rather than spatial variation in LUC accounted for the majority of variation in water quality. Urbanization and pasture development generally degraded water quality while agriculture and secondary forest re-growth had mixed impacts. The spatial scale over which LUC influenced water quality differed across indicators. Turbidity and dissolved oxygen (DO) responded to LUC in large-scale watersheds, in-stream nitrogen concentrations to LUC in riparian buffers of large watersheds, and fecal matter content and in-stream phosphorus concentration to LUC at the sub-watershed scale. Stream discharge modulated impacts of LUC on water quality for most of the metrics. Our findings highlight the importance of considering multiple spatial scales for understanding the impacts of human activities on watershed ecosystem services. PMID:26146455

A Genetic Approach to the Identification of Plant Genes Involved in Viral Movement

DTIC Science & Technology

1999-09-30

Arabidopsis.. We plan to create two transgenic plant lines of Arabidopsis, one that expresses the firefly luciferase (Luc) gene, and one that... transgenic plant lines that express Luc or CD upon infection with RCNMV. Phenotypically these lines will either be luminescent (Luc) or sensitive to 5

Posttranslationally caused bioluminescence burst of the Escherichia coli luciferase reporter strain.

PubMed

Ideguchi, Yamato; Oshikoshi, Yuta; Ryo, Masashi; Motoki, Shogo; Kuwano, Takashi; Tezuka, Takafumi; Aoki, Setsuyuki

2016-01-01

We continuously monitored bioluminescence from a wild-type reporter strain of Escherichia coli (lacp::luc+/WT), which carries the promoter of the lac operon (lacp) fused with the firefly luciferase gene (luc+). This strain showed a bioluminescence burst when shifted into the stationary growth phase. Bioluminescence profiles of other wild-type reporter strains (rpsPp::luc+ and argAp::luc+) and gene-deletion reporter strains (lacp::luc+/crp- and lacp::luc+/lacI-) indicate that transcriptional regulation is not responsible for generation of the burst. Consistently, changes in the luciferase protein levels did not recapitulate the profile of the burst. On the other hand, dissolved oxygen levels increased over the period across the burst, suggesting that the burst is, at least partially, caused by an increase in intracellular oxygen levels. We discuss limits of the firefly luciferase when used as a reporter for gene expression and its potential utility for monitoring metabolic changes in cells.

Monitoring autophagic flux using p62/SQSTM1 based luciferase reporters in glioma cells.

PubMed

Min, Zhang; Ting, Yao; Mingtao, Gong; Xiaofei, Tang; Dong, Yan; Chenguang, Zhang; Wei, Ding

2018-02-01

Autophagy is a highly dynamic process characterized with the term of autophagic flux. In the present study, we developed a quantifiable luciferase reporter system to measure the capacity as well as the dynamics of autophagic flux. Briefly, a luciferase variant of Luc2p was fused with p62/SQSTM1 or its UBA domain deletion mutant (p62ΔU) and transfected into cells. The expressed Luc2p-p62 fusion protein was primarily degraded via autophagy, while Luc2p-p62ΔU was employed as a normalization control due to its resistance to autophagic degradation. The luciferase activity of the lysates from two parallel populations of glioma cells expressing either Luc2p-p62 or Luc2p-p62ΔU was determined and the ratio of Luc2p-p62ΔU/Luc2p-p62 was used to assay the autophagic flux. By this approach, the induction of autophagy was manifested as an increased Luc2p-p62ΔU/Luc2p-p62 ratio, which could be neutralized by autophagy inhibitors or knockdown of ATG5. The performance of our autophagic flux detection system was comparable to a recently reported GFP-LC3-RFP-LC3ΔG probe. We tested the system in TMZ treated glioma cells, and found that coadministration of chloroquine to attenuate cellular autophagic flux significantly improved the TMZ efficacy by triggering more early apoptosis. Collectively, our luciferase-based autophagic flux assay may serve as a useful alternative yet sensitive method for autophagic flux detection in tumor cells. Copyright © 2018 Elsevier Inc. All rights reserved.

Optimized Replicating Renilla Luciferase Reporter HIV-1 Utilizing Novel Internal Ribosome Entry Site Elements for Native Nef Expression and Function.

PubMed

Alberti, Michael O; Jones, Jennifer J; Miglietta, Riccardo; Ding, Haitao; Bakshi, Rakesh K; Edmonds, Tara G; Kappes, John C; Ochsenbauer, Christina

2015-12-01

We previously developed replication-competent reporter HIV-1 (referred to herein as LucR.T2A reporter viruses), utilizing a "ribosome skipping" T2A peptide strategy to link Renilla luciferase (LucR) with Nef expression. The demonstrated utility for HIV-1 vaccine and transmission study applications included measurement of neutralizing antibody (NAb) activity in vaccine sera, improved cell-mediated virus inhibition assays, such as T cell-mediated virus inhibition and antibody-dependent cell-mediated cytotoxicity (ADCC) assays, and humanized mouse models. Herein, we extend our prior work and introduce reporter virus technology for applications that require fully functional Nef. We demonstrate that in CD4(+) T cells productively infected with LucR.T2A reporter viruses, T2A peptide-driven Nef expression and function, such as down-regulation of surface CD4 and MHC-I, were impaired. We overcame this limitation of LucR.T2A reporter viruses and achieved physiological Nef expression and function by engineering novel LucR reporter HIV-1 comprising 11 different internal ribosome entry site (IRES) elements chosen for size and relative activity. A range of Nef expression was observed in 293T cells transfected with the different LucR.IRES reporter virus constructs. Iteratively, we identified IRES reporter genomes that expressed Nef closest to physiological levels and produced virus with infectivity, titers, and replication kinetics similar to nonreporter viruses. Our results demonstrated that LucR reporter activity was stable over multiple replication cycles in peripheral blood mononuclear cells (PBMCs). Furthermore, we analyzed Nef functionality, i.e., down-modulation of MHC-I and CD4, following infection of T cell lines and PBMCs. Unlike LucR.T2A reporter virus, one of the redesigned LucR.IRES reporter viruses [containing the modified encephalomyocarditis virus (EMCV) 6ATR IRES element, "6ATRi"] demonstrated Nef expression and function similar to parental "nonreporter" virus

A Double Selection Approach to Achieve Specific Expression of Toxin Genes for Ovarian Cancer Gene Therapy

DTIC Science & Technology

2007-11-01

cells ( gray fill), cells preincubated with PBS and infected with virus (solid line), and cells preincubated with recombinant knob protein and incubated...U118-hCAR-tailless cells (b). Gray line¼ cells alone, solid line¼ cells+Ad5Luc1-CK1, dashed line¼ cells+Ad5Luc1. Figure 5 Ad5Luc1-CK1CAR-independent...line, whereas U118MG-hCAR-tailless stably expresses the extracellular domain of human CAR. Cells were infected with Ad5Luc1 ( gray bar) and Ad5-r1 (black

TAF1, From a General Transcription Factor to Modulator of Androgen Receptor in Prostate Cancer

DTIC Science & Technology

2008-02-01

or pARR3tk-Luc), and the Renilla luciferase vector (pRL-TK) as an internal control. Cells were then treated with or without R1881 for 24 h prior to...increasing amount of HA tagged full length TAF1 (pCS2+/TAF1), pRL-TK- Renilla (83 ng/well), and pARR3-tk-Luc (167 ng/well) (A) or pPSA-Luc (1ug/well...numbers of pRL-TK- Renilla , as a non androgenic reporter was also enhanced by TAF1 over-expression in both LNCaP and PC3 cells, the ARR3tk-Luc

Real-time monitoring of stress erythropoiesis in vivo using Gata1 and beta-globin LCR luciferase transgenic mice.

PubMed

Suzuki, Mikiko; Ohneda, Kinuko; Hosoya-Ohmura, Sakie; Tsukamoto, Saho; Ohneda, Osamu; Philipsen, Sjaak; Yamamoto, Masayuki

2006-07-15

Erythroid progenitors have the potential to proliferate rapidly in response to environmental stimuli. This process is referred to as stress erythropoiesis, with erythropoietin (EPO) playing central roles in its promotion. In this study, we wanted to elucidate the molecular mechanisms governing the regulation of stress erythropoiesis and the maintenance of red-cell homeostasis. This was achieved by our development of a noninvasive real-time monitoring system for erythropoiesis using transgenic mouse lines expressing luciferase under the control of the mouse Gata1 hematopoietic regulatory domain (G1-HRD-luc) or human beta-globin locus control region (Hbb-LCR-luc). Optical bioluminescence images revealed that the luciferase was specifically expressed in spleen and bone marrow and was induced rapidly in response to anemia and hypoxia stimuli. The G1-HRD-luc activity tracked the emergence and disappearance of proerythroblast-stage progenitors, whereas the Hbb-LCR-luc activity tracked erythroblasts and later stage erythroid cells. Increased plasma EPO concentration preceded an increase in G1-HRD-luc, supporting our contention that EPO acts as the key upstream signal in stress erythropoiesis. Hence, we conclude that G1-HRD-luc and Hbb-LCR-luc reporters are differentially activated during stress erythropoiesis and that the transgenic mouse lines used serve as an important means for understanding the homeostatic regulation of erythropoiesis.

Optimized Replicating Renilla Luciferase Reporter HIV-1 Utilizing Novel Internal Ribosome Entry Site Elements for Native Nef Expression and Function

PubMed Central

Alberti, Michael O.; Jones, Jennifer J.; Miglietta, Riccardo; Ding, Haitao; Bakshi, Rakesh K.; Edmonds, Tara G.; Kappes, John C.

2015-01-01

Abstract We previously developed replication-competent reporter HIV-1 (referred to herein as LucR.T2A reporter viruses), utilizing a “ribosome skipping” T2A peptide strategy to link Renilla luciferase (LucR) with Nef expression. The demonstrated utility for HIV-1 vaccine and transmission study applications included measurement of neutralizing antibody (NAb) activity in vaccine sera, improved cell-mediated virus inhibition assays, such as T cell-mediated virus inhibition and antibody-dependent cell-mediated cytotoxicity (ADCC) assays, and humanized mouse models. Herein, we extend our prior work and introduce reporter virus technology for applications that require fully functional Nef. We demonstrate that in CD4+ T cells productively infected with LucR.T2A reporter viruses, T2A peptide-driven Nef expression and function, such as down-regulation of surface CD4 and MHC-I, were impaired. We overcame this limitation of LucR.T2A reporter viruses and achieved physiological Nef expression and function by engineering novel LucR reporter HIV-1 comprising 11 different internal ribosome entry site (IRES) elements chosen for size and relative activity. A range of Nef expression was observed in 293T cells transfected with the different LucR.IRES reporter virus constructs. Iteratively, we identified IRES reporter genomes that expressed Nef closest to physiological levels and produced virus with infectivity, titers, and replication kinetics similar to nonreporter viruses. Our results demonstrated that LucR reporter activity was stable over multiple replication cycles in peripheral blood mononuclear cells (PBMCs). Furthermore, we analyzed Nef functionality, i.e., down-modulation of MHC-I and CD4, following infection of T cell lines and PBMCs. Unlike LucR.T2A reporter virus, one of the redesigned LucR.IRES reporter viruses [containing the modified encephalomyocarditis virus (EMCV) 6ATR IRES element, “6ATRi”] demonstrated Nef expression and function similar to parental �

Teaching Medical Student Psychiatry Through Contemporary Music

ERIC Educational Resources Information Center

Egan, William H.

1977-01-01

An audio technique uses contemporary music recordings to illustrate various personality disorders, including: schizoid, paranoid, compulsive, antisocial, and hysterical. The works of Bob Dylan, Paul Simon, Harry Chapin, the Beatles, Janis Ian, James Taylor, Tammy Wynette, and others are cited. (LBH)

Comparison of monkeypox viruses pathogenesis in mice by in vivo imaging

USGS Publications Warehouse

Osorio, Jorge E.; Iams, Keith P.; Meteyer, Carol U.; Rocke, Tonie E.

2009-01-01

Monkeypox viruses (MPXV) cause human monkeypox, a zoonotic smallpox-like disease endemic to Africa, and are of worldwide public health and biodefense concern. Using viruses from the Congo (MPXV-2003-Congo-358) and West African (MPXV-2003-USA-044) clades, we constructed recombinant viruses that express the luciferase gene (MPXV-Congo/Luc+and MPXV-USA-Luc+) and compared their viral infection in mice by biophotonic imaging. BALB/c mice became infected by both MPXV clades, but they recovered and cleared the infection within 10 days post-infection (PI). However, infection in severe combined immune deficient (SCID) BALB/c mice resulted in 100% lethality. Intraperitoneal (IP) injection of both MPXV-Congo and MPXV-Congo/Luc+resulted in a systemic clinical disease and the same mean time-to-death at 9 (??0) days post-infection. Likewise, IP injection of SCID-BALB/c mice with MPXV-USA or the MPXV-USA-Luc+, resulted in similar disease but longer (P<0.05) mean time-to-death (11??0 days) for both viruses compared to the Congo strains. Imaging studies in SCID mice showed luminescence in the abdomen within 24 hours PI with subsequent spread elsewhere. Animals infected with the MPXV-USA/Luc+had less intense luminescence in tissues than those inoculated with MPXV-Congo/Luc+, and systemic spread of the MPXV-USA/Luc+virus occurred approximately two days later than the MPXV-Congo/Luc+. The ovary was an important target for viral replication as evidenced by the high viral titers and immunohistochemistry. These studies demonstrate the suitability of a mouse model and biophotonic imaging to compare the disease progression and tissue tropism of MPX viruses.

Bioluminescent system for dynamic imaging of cell and animal behavior

DOE Office of Scientific and Technical Information (OSTI.GOV)

Hara-Miyauchi, Chikako; Laboratory for Cell Function Dynamics, Brain Science Institute, RIKEN, Saitama 351-0198; Department of Biophysics and Biochemistry, Graduate School of Health Care Sciences, Tokyo Medical and Dental University, Tokyo 113-8510

2012-03-09

Highlights: Black-Right-Pointing-Pointer We combined a yellow variant of GFP and firefly luciferase to make ffLuc-cp156. Black-Right-Pointing-Pointer ffLuc-cp156 showed improved photon yield in cultured cells and transgenic mice. Black-Right-Pointing-Pointer ffLuc-cp156 enabled video-rate bioluminescence imaging of freely-moving animals. Black-Right-Pointing-Pointer ffLuc-cp156 mice enabled tracking real-time drug delivery in conscious animals. -- Abstract: The current utility of bioluminescence imaging is constrained by a low photon yield that limits temporal sensitivity. Here, we describe an imaging method that uses a chemiluminescent/fluorescent protein, ffLuc-cp156, which consists of a yellow variant of Aequorea GFP and firefly luciferase. We report an improvement in photon yield by over threemore » orders of magnitude over current bioluminescent systems. We imaged cellular movement at high resolution including neuronal growth cones and microglial cell protrusions. Transgenic ffLuc-cp156 mice enabled video-rate bioluminescence imaging of freely moving animals, which may provide a reliable assay for drug distribution in behaving animals for pre-clinical studies.« less

Carbon Calculator for Land Use Change from Biofuels Production (CCLUB) Users’ Manual and Technical Documentation

DOE Office of Scientific and Technical Information (OSTI.GOV)

Dunn, Jennifer B.; Qin, Zhangcai; Mueller, Steffen

The Carbon Calculator for Land Use Change from Biofuels Production (CCLUB) calculates carbon emissions from land use change (LUC) for four different ethanol production pathways including corn grain ethanol and cellulosic ethanol from corn stover, Miscanthus, and switchgrass, and a soy biodiesel pathway. This document discusses the version of CCLUB released September 30, 2017 which includes five ethanol LUC scenarios and four soy biodiesel LUC scenarios.

Selected Eastern Europe Sociological Translations

DTIC Science & Technology

1960-02-29

direction» There teachers Kujtim Dedej, Jani Xhupir Antulla Mosko, etc., conduct practical laboratory experiments in chemistry, physics, mathematics...and Pogradec have done excellent work’ in this respect. However, the gymnasium of Gjinokaster, under the direction of Kujtim Dejej, has done

Assessment of malaria transmission changes in Africa, due to the climate impact of land use change using Coupled Model Intercomparison Project Phase 5 earth system models.

PubMed

Tompkins, Adrian M; Caporaso, Luca

2016-03-31

Using mathematical modelling tools, we assessed the potential for land use change (LUC) associated with the Intergovernmental Panel on Climate Change low- and high-end emission scenarios (RCP2.6 and RCP8.5) to impact malaria transmission in Africa. To drive a spatially explicit, dynamical malaria model, data from the four available earth system models (ESMs) that contributed to the LUC experiment of the Fifth Climate Model Intercomparison Project are used. Despite the limited size of the ESM ensemble, stark differences in the assessment of how LUC can impact climate are revealed. In three out of four ESMs, the impact of LUC on precipitation and temperature over the next century is limited, resulting in no significant change in malaria transmission. However, in one ESM, LUC leads to increases in precipitation under scenario RCP2.6, and increases in temperature in areas of land use conversion to farmland under both scenarios. The result is a more intense transmission and longer transmission seasons in the southeast of the continent, most notably in Mozambique and southern Tanzania. In contrast, warming associated with LUC in the Sahel region reduces risk in this model, as temperatures are already above the 25-30°C threshold at which transmission peaks. The differences between the ESMs emphasise the uncertainty in such assessments. It is also recalled that the modelling framework is unable to adequately represent local-scale changes in climate due to LUC, which some field studies indicate could be significant.

77 FR 62490 - Performance Review Board Appointments

Federal Register 2010, 2011, 2012, 2013, 2014

2012-10-15

...; Gutter, Karis T.; Harden, Krysta.; Hipp, Janie; Holtzman, Max T.; Jett, Carole E.; Jones, Carmen....; Chasteen, G. Taylor; Christian, Lisa A.; Clanton, Michael W.; Coffee, Richard; Farington, Kim S.; Foster....; Watts, Michael; White, John S.; White, Sharmian L.; Wilburn, Curtis; Wilusz, Lisa; Young, Benjamin...

76 FR 67402 - Performance Review Board Appointments

Federal Register 2010, 2011, 2012, 2013, 2014

2011-11-01

....; Harden, Krysta; Hipp, Janie; Holtzman, Max T.; Jett, Carole E.; MacMillian, Anne; Mande, Jerold; Mills...; Christian, Lisa A.; Clanton, Michael W.; Davenport, Peter; Douglas, Walt; Foster, Andrea L.; Golden, John....; White, Sharmian L.; Wilburn, Curtis; Wilusz, Lisa; Worthington, Ruth M.; Young, Benjamin; Young, Mike...

Proceedings of the Annual Meeting of the Association for Education in Journalism and Mass Communication (86th, Kansas City, Missouri, July 30-August 2, 2003). Miscellaneous Divisions.

ERIC Educational Resources Information Center

2003

The Miscellaneous Divisions of the proceedings contains the following 17 papers: "Analyzing Sequential Art: Visual Narrative Techniques in 'Calvin and Hobbes'" (Sharron M. Hope); "A Critical Vision of Gender in 2002 Campaign Ads" (Janis Teruggi Page); "Personal Impact Assessment of Advertising Culture of 'Whiteness':…

The Bioluminescence Resonance Energy Transfer from Firefly Luciferase to a Synthetic Dye and its Application for the Rapid Homogeneous Immunoassay of Progesterone.

PubMed

Smirnova, Daria V; Samsonova, Jeanne V; Ugarova, Natalia N

2016-01-01

The sensitive BRET system for the homogeneous immunoassay of a low-molecular weight antigen was developed using progesterone as an example. Two thermostable mutants of the Luciola mingrelica firefly luciferase (Luc)-the "red" mutant with λmax.em = 590 nm (RedLuc) and the "green" mutant with λmax.em = 550 nm (GreenLuc)-were tested as the donors. The water-soluble Alexa Fluor 610× (AF) dye was selected as the acceptor because its two absorption maxima, located at 550 and 610 nm, are close to the bioluminescence maxima of the GreenLuc and RedLuc, respectively. The methods for the synthesis of the luciferase-progesterone (Luc-Pg) conjugate and the conjugate of the dye and the polyclonal antiprogesterone antibody (AF-Ab) were developed. Both conjugates retained their functional properties, had high antigen-antibody binding activity, and demonstrated a high BRET signal. The homogeneous immunoassay system based on the BRET from the firefly luciferase to the synthetic dye was established to assay progesterone as a model antigen. Optimization of the assay conditions, the composition of the reaction mixture, and the concentrations of the donor and the acceptor made it possible to reach the minimum detectable progesterone concentration of 0.5 ng mL(-1) . © 2015 The American Society of Photobiology.

Characterization of luciferases and its paralogue in the Panamanian luminous click beetle Pyrophorus angustus: a click beetle luciferase lacks the fatty acyl-CoA synthetic activity.

PubMed

Oba, Yuichi; Kumazaki, Mizuho; Inouye, Satoshi

2010-02-15

Two luciferase genes (dPaLuc and vPaLuc) and one paralogue of luciferase (PaLL) were isolated from the Panamanian luminous click beetle, Pyrophorus angustus (Elateridae, Pyrophorinae). The transcripts of dPaLuc and vPaLuc were predominantly detected in the body parts with dorsal photophore and ventral photophore, respectively, and the transcript of PaLL was detected in both parts. The gene products of dPaLuc and vPaLuc possessed luminescence activity with firefly luciferin (lambda(max)=536 and 566 nm, respectively) but did not show significant activity of fatty acyl-CoA synthesis. On the other hand, the gene product of PaLL had fatty acyl-CoA synthetic activity with very weak luminescence activity. The catalytic properties of click beetle luciferase are different from our previous results that firefly luciferase has both luminescence activity and fatty acyl-CoA synthetic activity. These results suggested that the ancestral fatty acyl-CoA synthetase in the Pyrophorinae lineage has undergone gene duplication event, followed by specialization of one copy in luciferase. Subsequently, the luciferase was duplicated again and the two copies diverged in their luminescent color and expression pattern. Copyright 2009 Elsevier B.V. All rights reserved.

Nanodiamond modified copolymer scaffolds affects tumour progression of early neoplastic oral keratinocytes.

PubMed

Suliman, Salwa; Mustafa, Kamal; Krueger, Anke; Steinmüller-Nethl, Doris; Finne-Wistrand, Anna; Osdal, Tereza; Hamza, Amani O; Sun, Yang; Parajuli, Himalaya; Waag, Thilo; Nickel, Joachim; Johannessen, Anne Christine; McCormack, Emmet; Costea, Daniela Elena

2016-07-01

This study aimed to evaluate the tumorigenic potential of functionalising poly(LLA-co-CL) scaffolds. The copolymer scaffolds were functionalised with nanodiamonds (nDP) or with nDP and physisorbed BMP-2 (nDP-PHY) to enhance osteoinductivity. Culturing early neoplastic dysplastic keratinocytes (DOK(Luc)) on nDP modified scaffolds reduced significantly their subsequent sphere formation ability and decreased significantly the cells' proliferation in the supra-basal layers of in vitro 3D oral neoplastic mucosa (3D-OT) when compared to DOK(Luc) previously cultured on nDP-PHY scaffolds. Using an in vivo non-invasive environmentally-induced oral carcinogenesis model, nDP scaffolds were observed to reduce bioluminescence intensity of tumours formed by DOK(Luc) + carcinoma associated fibroblasts (CAF). nDP modification was also found to promote differentiation of DOK(Luc) both in vitro in 3D-OT and in vivo in xenografts formed by DOK(Luc) alone. The nDP-PHY scaffold had the highest number of invasive tumours formed by DOK(Luc) + CAF outside the scaffold area compared to the nDP and control scaffolds. In conclusion, in vitro and in vivo results presented here demonstrate that nDP modified copolymer scaffolds are able to decrease the tumorigenic potential of DOK(Luc), while confirming concerns for the therapeutic use of BMP-2 for reconstruction of bone defects in oral cancer patients due to its tumour promoting capabilities. Copyright © 2016 The Authors. Published by Elsevier Ltd.. All rights reserved.

Information Technology Challenges Facing the Strategic Leaders of Homeland Security in the 21st Century

DTIC Science & Technology

2004-05-03

A special thanks to all of these folks (Glenn Starnes, Lee Gutierrez, Bill Rapp, Joel Hillison, Carlos Gomez, Steve Fraunfelter, Janie Hopkins, Dave ...DiClemente, Joe Nunez, John Bonin , and Steve Nerheim). viii INFORMATION TECHNOLOGY CHALLENGES FACING THE STRATEGIC LEADERS OF HOMELAND SECURITY IN THE 21ST

ACUTE AND CHRONIC TOXICITY OF BREVETOXIN TO OYSTERS AND GRASS SHRIMP

EPA Science Inventory

Walker, Calvin C., James T. Winstead, Steven S. Foss, Janis C. Kurtz, James Watts, Jeanne E. Scott and William S. Fisher. In press. Acute and Chronic Toxicity of Brevetoxin to Oysters and Grass Shrimp (Abstract). To be presented at the SETAC Fourth World Congress, 14-18 November ...

Environmentally Induced Gene Silencing in Breast Cancer

DTIC Science & Technology

2007-07-01

fibrosarcoma cell line (HTD114), and a human breast cancer cell line (MCF7). The MLH1 promoter was only tested in the MCG7 cells. The control TRE-Luc...TRE- Luc MLH1 - Luc step in silencing is quite unstable. Nonetheless, cells that exhibit stable silencing of the HPRT construct can arise in...mechanism (i.e., gene repression). Finally, during the last year we have isolated or acquired functional promoters for the BRCA-1, MLH1 , and E

Replication Competent Molecular Clones of HIV-1 Expressing Renilla Luciferase Facilitate the Analysis of Antibody Inhibition in PBMC

PubMed Central

Edmonds, Tara G.; Ding, Haitao; Yuan, Xing; Wei, Qing; Smith, Kendra S.; Conway, Joan A.; Wieczorek, Lindsay; Brown, Bruce; Polonis, Victoria; West, John T.; Montefiori, David C.; Kappes, John C.; Ochsenbauer, Christina

2010-01-01

Effective vaccine development for human immunodeficiency virus type 1 (HIV-1) will require assays that ascertain the capacity of vaccine immunogens to elicit neutralizing antibodies (NAb) to diverse HIV-1 strains. To facilitate NAb assessment in peripheral blood mononuclear cell (PBMC)-based assays, we developed an assay-adaptable platform based on a Renilla luciferase (LucR) expressing HIV-1 proviral backbone. LucR was inserted into pNL4-3 DNA, preserving all viral open reading frames. The proviral genome was engineered to facilitate expression of diverse HIV-1 env sequences, allowing analysis in an isogenic background. The resulting Env-IMC-LucR viruses are infectious, and LucR is stably expressed over multiple replications in PBMC. HIV-1 neutralization, targeting TZM-bl cells, was highly correlative comparing virus (LucR) and cell (firefly luciferase) readouts. In PBMC, NAb activity can be analyzed either within a single or multiple cycles of replication. These results represent advancement toward a standardizable PBMC-based neutralization assay for assessing HIV-1 vaccine immunogen efficacy. PMID:20863545

Replication competent molecular clones of HIV-1 expressing Renilla luciferase facilitate the analysis of antibody inhibition in PBMC.

PubMed

Edmonds, Tara G; Ding, Haitao; Yuan, Xing; Wei, Qing; Smith, Kendra S; Conway, Joan A; Wieczorek, Lindsay; Brown, Bruce; Polonis, Victoria; West, John T; Montefiori, David C; Kappes, John C; Ochsenbauer, Christina

2010-12-05

Effective vaccine development for human immunodeficiency virus type 1 (HIV-1) will require assays that ascertain the capacity of vaccine immunogens to elicit neutralizing antibodies (NAb) to diverse HIV-1 strains. To facilitate NAb assessment in peripheral blood mononuclear cell (PBMC)-based assays, we developed an assay-adaptable platform based on a Renilla luciferase (LucR) expressing HIV-1 proviral backbone. LucR was inserted into pNL4-3 DNA, preserving all viral open reading frames. The proviral genome was engineered to facilitate expression of diverse HIV-1 env sequences, allowing analysis in an isogenic background. The resulting Env-IMC-LucR viruses are infectious, and LucR is stably expressed over multiple replications in PBMC. HIV-1 neutralization, targeting TZM-bl cells, was highly correlative comparing virus (LucR) and cell (firefly luciferase) readouts. In PBMC, NAb activity can be analyzed either within a single or multiple cycles of replication. These results represent advancement toward a standardizable PBMC-based neutralization assay for assessing HIV-1 vaccine immunogen efficacy. Copyright © 2010 Elsevier Inc. All rights reserved.

Payback time for soil carbon and sugar-cane ethanol

NASA Astrophysics Data System (ADS)

Mello, Francisco F. C.; Cerri, Carlos E. P.; Davies, Christian A.; Holbrook, N. Michele; Paustian, Keith; Maia, Stoécio M. F.; Galdos, Marcelo V.; Bernoux, Martial; Cerri, Carlos C.

2014-07-01

The effects of land-use change (LUC) on soil carbon (C) balance has to be taken into account in calculating the CO2 savings attributed to bioenergy crops. There have been few direct field measurements that quantify the effects of LUC on soil C for the most common land-use transitions into sugar cane in Brazil, the world's largest producer . We quantified the C balance for LUC as a net loss (carbon debt) or net gain (carbon credit) in soil C for sugar-cane expansion in Brazil. We sampled 135 field sites to 1 m depth, representing three major LUC scenarios. Our results demonstrate that soil C stocks decrease following LUC from native vegetation and pastures, and increase where cropland is converted to sugar cane. The payback time for the soil C debt was eight years for native vegetation and two to three years for pastures. With an increasing need for biofuels and the potential for Brazil to help meet global demand, our results will be invaluable for guiding expansion policies of sugar-cane production towards greater sustainability.

Effects of Postchallenge Administration of ST-246 on Dissemination of IHD-J-Luc Vaccinia Virus in Normal Mice and in Immune-Deficient Mice Reconstituted with T Cells

PubMed Central

Shotwell, Elisabeth; Scott, John; Cruz, Stephanie; King, Lisa R.; Manischewitz, Jody; Diaz, Claudia G.; Jordan, Robert A.; Grosenbach, Douglas W.; Golding, Hana

2013-01-01

Whole-body bioimaging was used to study dissemination of vaccinia virus (VACV) in normal and in immune deficient (nu−/nu−) mice protected from lethality by postchallenge administration of ST-246. Total fluxes were recorded in the liver, spleen, lungs, and nasal cavities of live mice after intranasal infection with a recombinant IHD-J-Luc VACV expressing luciferase. Areas under the flux curve were calculated for individual mice to assess viral loads. Treatment for 2 to 5 days of normal BALB/c mice with ST-246 at 100 mg/kg starting 24 h postchallenge conferred 100% protection and reduced viral loads in four organs compared to control mice. Mice also survived after 5 days of treatment with ST-246 at 30 mg/kg, and yet the viral loads and poxes were higher in these mice compared to 100-mg/kg treatment group. Nude mice were not protected by ST-246 alone or by 10 million adoptively transferred T cells. In contrast, nude mice that received T cells and 7-day treatment with ST-246 survived infection and exhibited reduced viral loads compared to nonreconstituted and ST-246-treated mice after ST-246 was stopped. Similar protection of nude mice was achieved using adoptively transferred 1.0 and 0.1 million, but not 0.01 million, purified T cells or CD4+ or CD8+ T cells in conjunction with ST-246 treatment. These data suggest that ST-246 protects immunocompetent mice from lethality and reduces viral dissemination in internal organs and poxvirus lesions. Furthermore, immune-deficient animals with partial T cell reconstitution can control virus replication after a course of ST-246 and survive lethal vaccinia virus challenge. PMID:23468500

Human Adipose Tissue Stem Cells Promote the Growth of Acute Lymphoblastic Leukemia Cells in NOD/SCID Mice.

PubMed

Lee, Myoung Woo; Park, Yoo Jin; Kim, Dae Seong; Park, Hyun Jin; Jung, Hye Lim; Lee, Ji Won; Sung, Ki Woong; Koo, Hong Hoe; Yoo, Keon Hee

2018-06-01

In this study, the effect of adipose tissue stem cells (ASCs) on the growth of acute lymphoblastic leukemia (ALL) cells was examined in an in vivo model. We established ALL cell lines expressing firefly luciferase (ALL/fLuc) by lentiviral infection that were injected intraperitoneally to NOD/SCID mice. The luciferase activities were significantly higher in mice co-injected with 10 5 ALL/fLuc cells and ASCs than in those injected with ALL/fLuc cells alone. Co-injection of 10 5 ALL/fLuc cells and ASCs in differing ratios into mice gradually increased the bioluminescence intensity in all groups, and mice co-injected with 1 or 2 × 10 6 ASCs showed higher bioluminescence intensity than those receiving lower numbers. Interestingly, in the mice injected with 10 5 or 10 7 ALL/fLuc cells alone, the formation of tumor masses was not observed for at least five weeks. Moreover, co-injection of 10 7 ALL/fLuc cells and 5 × 10 5 ASCs into mice increased the bioluminescence intensity in all groups, and showed significantly higher bioluminescence intensity compared to mice co-injected with human normal fibroblast HS68 cells. Overall, ASCs promote the growth of ALL cells in vivo, suggesting that ASCs negatively influence hematologic malignancy, which should be considered in developing cell therapy using ASCs.

Effects of Organic and Inorganic Fertilizers on Marigold Growth and Flowering

USDA-ARS?s Scientific Manuscript database

Two experiments were conducted to evaluate the growth and flowering responses of greenhouse-grown French marigold (Tagetes patula L. ‘Janie Deep Orange’) to two non-composted broiler chicken litter-based organic fertilizers 4-2-2 and 3-3-3, and one commonly used synthetic controlled-release fertiliz...

Effective Teaching in the Multi-Cultural Classroom.

ERIC Educational Resources Information Center

Gotch, Donna; Brydges, Michael

Community college instructors and administrators need to understand teacher immediacy research and the role of immediacy in the multi-cultural classroom. Immediacy can be viewed as a combination of nonverbal behaviors used to accentuate a verbal message and reduce physical and psychological distance between interactants. Janis Andersen's research…

Small Group Dynamics and the Watergate Coverup: A Case Study of Groupthink.

ERIC Educational Resources Information Center

Cline, Rebecca J.

The decisions President Richard Nixon and his closest advisors made in the Watergate coverup were products of what Irving Janis calls "groupthink." Groupthink, a type of decision-making emphasizing unanimity over objective evaluation, develops when the decision makers (1) form a group of marked cohesiveness, (2) insulate themselves from…

Decisionmaking Context Model for Enhancing Evaluation Utilization.

ERIC Educational Resources Information Center

Brown, Robert D.; And Others

1984-01-01

This paper discusses two models that hold promise for helping evaluators understand and cope with different decision contexts: (1) the conflict Model (Janis and Mann, 1977) and the Social Process Model (Vroom and Yago, 1974). Implications and guidelines for using decisionmaking models in evaluation settings are presented. (BS)

Prescription of medicated feedingstuffs.

PubMed

McDonald, Janis

2012-08-11

This article, by Janis McDonald of the Veterinary Medicines Directorate (VMD), sets out advice on best practice in using medicated feedingstuffs prescriptions. It has been produced by the VMD in the light of concerns that procedures for completing medicated feedingstuffs prescriptions may not always be being followed correctly.

Defining Strong State Accountability Systems: How Can Better Standards Gain Greater Traction? A First Look

ERIC Educational Resources Information Center

Reed, Eileen; Scull, Janie; Slicker, Gerilyn; Winkler, Amber M.

2012-01-01

Rigorous standards and aligned assessments are vital tools for boosting education outcomes but they have little traction without strong accountability systems that attach consequences to performance. In this pilot study, Eileen Reed, Janie Scull, Gerilyn Slicker, and Amber Winkler lay out the essential features of such accountability systems,…

Multimodal Imaging of Pathophysiological Changes and Their Role in Development of Breast Cancer Brain Metastasis

DTIC Science & Technology

2012-09-01

and tissue oxygenation. Moreover, by introducing hypoxia reporter gene ( HRE -luciferase) into breast tumor lines, we will be able to use...hypoxia reporter gene, HRE -ODD-luc. The single nodule lesion was visualized and followed up by both BLI and MRI. As an example presented in Figure 1...MDA-MB231 cells with stable transfection of a hypoxia reporter gene, HRE -ODD-luc. a. 3 × 105 MDA-MB231/5HRE-ODD-luc cells incubated in each well of

"Feminism Lite?" Feminist Identification, Speaker Appearance, and Perceptions of Feminist and Antifeminist Messengers

ERIC Educational Resources Information Center

Bullock, Heather E.; Fernald, Julian L.

2003-01-01

Drawing on a communications model of persuasion (Hovland, Janis, & Kelley, 1953), this study examined the effect of target appearance on feminists' and nonfeminists' perceptions of a speaker delivering a feminist or an antifeminist message. One hundred three college women watched one of four videotaped speeches that varied by content (profeminist…

15. 'Southern Pacific R.R., One 236'0' Bet. End Pins S. ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

15. 'Southern Pacific R.R., One 236'-0' Bet. End Pins S. Tr. Thro'. Draw Bridge over Sacramento River at Tehama, The Phoenix Bridge Co., Phoenixville, Pa., Jan'y 19th 98, Dwg. 585.' - Southern Pacific Railroad Shasta Route, Bridge No. 210.52, Milepost 210.52, Tehama, Tehama County, CA

Harvard Education Letter. Volume 20, Number 5, September-October 2004

ERIC Educational Resources Information Center

Sadowski, Michael, Ed.

2004-01-01

"Harvard Education Letter" is published bimonthly at the Harvard Graduate School of Education. This issue of "Harvard Education Letter" contains the following articles: (1) Adolescent Literacy: Are We Overlooking the Struggling Teenage Reader? (Robert Rothman); (2) The "N-Word" and the Racial Dynamics of Teaching (Wendy Luttrell and Janie Ward);…

Regulation of Kruppel-like factor 4, 9, and 13 genes and the steroidogenic genes LDLR, StAR, and CYP11A in ovarian granulosa cells.

PubMed

Natesampillai, Sekar; Kerkvliet, Jason; Leung, Peter C K; Veldhuis, Johannes D

2008-02-01

Kruppel-like factors (KLFs) are important Sp1-like eukaryotic transcriptional proteins. The LDLR, StAR, and CYP11A genes exhibit GC-rich Sp1-like sites, which have the potential to bind KLFs in multiprotein complexes. We now report that KLF4, KLF9, and KLF13 transcripts are expressed in and regulate ovarian cells. KLF4 and 13, but not KLF9, mRNA expression was induced and then repressed over time (P < 0.001). Combined LH and IGF-I stimulation increased KLF4 mRNA at 2 h (P < 0.01), whereas LH decreased KLF13 mRNA at 6 h (P < 0.05), and IGF-I reduced KLF13 at 24 h (P < 0.01) compared with untreated control. KLF9 was not regulated by either hormone. Transient transfection of KLF4, KLF9, and KLF13 suppressed LDLR/luc, StAR/luc, and CYP11A/luc by 80-90% (P < 0.001). Histone-deacetylase (HDAC) inhibitors stimulated LDLR/luc five- to sixfold and StAR/luc and CYP11A/luc activity twofold (P < 0.001) and partially reversed suppression by all three KLFs (P < 0.001). Deletion of the zinc finger domain of KLF13 abrogated repression of LDLR/luc. Lentiviral overexpression of the KLF13 gene suppressed LDLR mRNA (P < 0.001) and CYP11A mRNA (P = 0.003) but increased StAR mRNA (P = 0.007). Collectively, these data suggest that KLFs may recruit inhibitory complexes containing HDAC corepressors, thereby repressing LDLR and CYP11A transcription. Conversely, KLF13 may recruit unknown coactivators or stabilize StAR mRNA, thereby explaining enhancement of in situ StAR gene expression. These data introduce new potent gonadal transregulators of genes encoding proteins that mediate sterol uptake and steroid biosynthesis.

Multifunctional triblock co-polymer mP3/4HB-b-PEG-b-lPEI for efficient intracellular siRNA delivery and gene silencing.

PubMed

Zhou, Li; Chen, Zhifei; Wang, Feifei; Yang, Xiuqun; Zhang, Biliang

2013-04-01

A non-viral siRNA carrier composed of mono-methoxy-poly (3-hydroxybutyrate-co-4-hydroxybutyrate)-block-polyethylene glycol-block-linear polyethyleneimine (mP3/4HB-b-PEG-b-lPEI) was synthesized using 1800 Da linear polyethyleneimine and evaluated for siRNA delivery. Our study demonstrated that siRNA could be efficiently combined with mP3/4HB-b-PEG-b-lPEI (mAG) co-polymer and was protected from nuclease degradation. The combined siRNA were released from the complexes easily under heparin competition. The particle size of the mAG/siRNA complexes was 158 nm, with a ζ-potential of around 28 mV. Atomic force microscopy images displayed spherical and homogeneously distributed complexes. The mAG block co-polymer displayed low cytotoxicity and efficient cellular uptake of Cy3-siRNA in A549 cells by flow cytometry and confocal microscopy. In vitro transfection efficiency of the block co-polymer was assessed using siRNA against luciferase in cultured A549-Luc, HeLa-Luc, HLF-Luc, A375-Luc and MCF-7-Luc cells. A higher transfection efficiency and lower cytotoxicity was obtained by mAG block co-polymer in five cell lines. Furthermore, a remarkable improvement in luciferase gene silencing efficiency of the mAG complex (up to 90-95%) over that of Lipofectamine™ 2000 (70-82%) was observed in HLF-Luc and A375-Luc cells. Additionally, a mAG/p65-siRNA complex also showed a better capability than Lipofectamine™ 2000/p65-siRNA complex to drastically reduce the p65 mRNA level down to 10-16% in HeLa, U251 and HUVEC cells at an N/P ratio of 70. Crown Copyright © 2013. Published by Elsevier Ltd. All rights reserved.

Establishment and characterization of in vivo orthotopic bioluminescent xenograft models from human osteosarcoma cell lines in Swiss nude and NSG mice.

PubMed

Marques da Costa, Maria Eugenia; Daudigeos-Dubus, Estelle; Gomez-Brouchet, Anne; Bawa, Olivia; Rouffiac, Valerie; Serra, Massimo; Scotlandi, Katia; Santos, Conceição; Geoerger, Birgit; Gaspar, Nathalie

2018-03-01

Osteosarcoma is one of the most common primary bone tumors in childhood and adolescence. Metastases occurrence at diagnosis or during disease evolution is the main therapeutic challenge. New drug evaluation to improve patient survival requires the development of various preclinical models mimicking at best the complexity of the disease and its metastatic potential. We describe here the development and characteristics of two orthotopic bioluminescent (Luc/mKate2) cell-derived xenograft (CDX) models, Saos-2-B-Luc/mKate2-CDX and HOS-Luc/mKate2-CDX, in different immune (nude and NSG mouse strains) and bone (intratibial and paratibial with periosteum activation) contexts. IVIS SpectrumCT system allowed both longitudinal computed tomography (CT) and bioluminescence real-time follow-up of primary tumor growth and metastatic spread, which was confirmed by histology. The murine immune context influenced tumor engraftment, primary tumor growth, and metastatic spread to lungs, bone, and spleen (an unusual localization in humans). Engraftment in NSG mice was found superior to that found in nude mice and intratibial bone environment more favorable to engraftment compared to paratibial injection. The genetic background of the two CDX models also led to distinct primary tumor behavior observed on CT scan. Saos-2-B-Luc/mKate2-CDX showed osteocondensed, HOS-Luc/mKate2-CDX osteolytic morphology. Bioluminescence defined a faster growth of the primary tumor and metastases in Saos-2-B-Luc/mKate2-CDX than in HOS-Luc/mKate2-CDX. The early detection of primary tumor growth and metastatic spread by bioluminescence allows an improved exploration of osteosarcoma disease at tumor progression, and metastatic spread, as well as the evaluations of anticancer treatments. Our orthotopic models with metastatic spread bring complementary information to other types of existing osteosarcoma models. © 2018 The Authors. Cancer Medicine published by John Wiley & Sons Ltd.

A spatial modeling framework to evaluate domestic biofuel-induced potential land use changed and emissions

USGS Publications Warehouse

Elliot, Joshua; Sharma, Bhavna; Best, Neil; Glotter, Michael; Dunn, Jennifer B.; Foster, Ian; Miguez, Fernando; Mueller, Steffen; Wang, Michael

2014-01-01

We present a novel bottom-up approach to estimate biofuel-induced land-use change (LUC) and resulting CO2 emissions in the U.S. from 2010 to 2022, based on a consistent methodology across four essential components: land availability, land suitability, LUC decision-making, and induced CO2 emissions. Using highresolution geospatial data and modeling, we construct probabilistic assessments of county-, state-, and national-level LUC and emissions for macroeconomic scenarios. We use the Cropland Data Layer and the Protected Areas Database to characterize availability of land for biofuel crop cultivation, and the CERES-Maize and BioCro biophysical crop growth models to estimate the suitability (yield potential) of available lands for biofuel crops. For LUC decisionmaking, we use a county-level stochastic partial-equilibrium modeling framework and consider five scenarios involving annual ethanol production scaling to 15, 22, and 29 BG, respectively, in 2022, with corn providing feedstock for the first 15 BG and the remainder coming from one of two dedicated energy crops. Finally, we derive high-resolution above-ground carbon factors from the National Biomass and Carbon Data set to estimate emissions from each LUC pathway. Based on these inputs, we obtain estimates for average total LUC emissions of 6.1, 2.2, 1.0, 2.2, and 2.4 gCO2e/MJ for Corn-15 Billion gallons (BG), Miscanthus × giganteus (MxG)-7 BG, Switchgrass (SG)-7 BG, MxG-14 BG, and SG-14 BG scenarios, respectively.

Tropical Land Use Conversion Effects on Soil Microbial Community Structure and Function: Emerging Patterns and Knowledge Gaps

NASA Astrophysics Data System (ADS)

Seeley, M.; Marin-Spiotta, E.

2016-12-01

Modifications in vegetation due to land use conversions (LUC) between primary forests, pasture, cropping systems, tree plantations, and secondary forests drive shifts in soil microbial communities. These microbial community alterations affect carbon sequestration, nutrient cycling, aboveground biomass, and numerous other soil processes. Despite their importance, little is known about soil microbial organisms' response to LUC, especially in tropical regions where LUC rates are greatest. This project identifies current trends and uncertainties in tropical soil microbiology by comparing 56 published studies on LUC in tropical regions. This review indicates that microbial biomass and functional groups shifted in response to LUC, supporting demonstrated trends in changing soil carbon stocks due to LUC. Microbial biomass was greatest in primary forests when compared to secondary forests and in all forests when compared to both cropping systems and tree plantations. No trend existed when comparing pasture systems and forests, likely due to variations in pasture fertilizer use. Cropping system soils had greater gram positive and less gram negative bacteria than forest soils, potentially resulting in greater respiration of older carbon stocks in agricultural soils. Bacteria dominated primary forests while fungal populations were greatest in secondary forests. To characterize changes in microbial communities resulting from land use change, research must reflect the biophysical variation across the tropics. A chi-squared test revealed that the literature sites represented mean annual temperature variation across the tropics (p-value=0.66).

Comparative analysis of distribution and abundance of West Nile and Eastern Equine Encephalomyelitis virus vectors in Suffolk County, New York, using human population density and land use/cover data

USGS Publications Warehouse

Rochlin, I.; Harding, K.; Ginsberg, H.S.; Campbell, S.R.

2008-01-01

Five years of CDC light trap data from Suffolk County, NY, were analyzed to compare the applicability of human population density (HPD) and land use/cover (LUC) classification systems to describe mosquito abundance and to determine whether certain mosquito species of medical importance tend to be more common in urban (defined by HPD) or residential (defined by LUC) areas. Eleven study sites were categorized as urban or rural using U.S. Census Bureau data and by LUC types using geographic information systems (GISs). Abundance and percent composition of nine mosquito taxa, all known or potential vectors of arboviruses, were analyzed to determine spatial patterns. By HPD definitions, three mosquito species, Aedes canadensis (Theobald), Coquillettidia perturbans (Walker), and Culiseta melanura (Coquillett), differed significantly between habitat types, with higher abundance and percent composition in rural areas. Abundance and percent composition of these three species also increased with freshwater wetland, natural vegetation areas, or a combination when using LUC definitions. Additionally, two species, Ae. canadensis and Cs. melanura, were negatively affected by increased residential area. One species, Aedes vexans (Meigen), had higher percent composition in urban areas. Two medically important taxa, Culex spp. and Aedes triseriatus (Say), were proportionally more prevalent in residential areas by LUC classification, as was Aedes trivittatus (Coquillett). Although HPD classification was readily available and had some predictive value, LUC classification resulted in higher spatial resolution and better ability to develop location specific predictive models.

Variations of Luzon Undercurrent from observations and numerical model simulations

NASA Astrophysics Data System (ADS)

Wang, Qingye; Zhai, Fangguo; Hu, Dunxin

2014-06-01

Significant intraseasonal variability (ISV) of about 45-80 days and seasonal variation of the Luzon Undercurrent (LUC) at 18°N are studied using direct current measurements and a high-resolution global Hybrid Coordinate Ocean Model. The variations of the LUC are vertically coherent with those of Kuroshio Current both on intraseasonal and seasonal time scales. The ISV of the LUC is dominated by eddies with diameters of about 200-300 km and extending from sea surface to intermediate layer east of Luzon Island. The LUC becomes strong (weak) when cyclonic (anticyclonic) eddies occur. The eddies east of Luzon Island mainly originate from the bifurcation point (˜13°N) of the North Equatorial Current. These eddies propagate northwestward at a typical propagation speed of about 0.16 m s-1 along the east coast of Philippines, gradually strengthen and pass the Luzon coast, and continue northward to Luzon strait. On seasonal time scale, the LUC is strong (weak) in boreal winter (summer), and this variation is related to the seasonal evolution of large-scale ocean circulation east of Philippines mainly controlled by local wind forcing.

Effect of land-use change and management on biogenic volatile organic compound emissions--selecting climate-smart cultivars.

PubMed

Rosenkranz, Maaria; Pugh, Thomas A M; Schnitzler, Jörg-Peter; Arneth, Almut

2015-09-01

Land-use change (LUC) has fundamentally altered the form and function of the terrestrial biosphere. Increasing human population, the drive for higher living standards and the potential challenges of mitigating and adapting to global environmental change mean that further changes in LUC are unavoidable. LUC has direct consequences on climate not only via emissions of greenhouse gases and changing the surface energy balance but also by affecting the emission of biogenic volatile organic compounds (BVOCs). Isoprenoids, which dominate global BVOC emissions, are highly reactive and strongly modify atmospheric composition. The effects of LUC on BVOC emissions and related atmospheric chemistry have been largely ignored so far. However, compared with natural ecosystems, most tree species used in bioenergy plantations are strong BVOC emitters, whereas intensively cultivated crops typically emit less BVOCs. Here, we summarize the current knowledge on LUC-driven BVOC emissions and how these might affect atmospheric composition and climate. We further discuss land management and plant-breeding strategies, which could be taken to move towards climate-friendly BVOC emissions while simultaneously maintaining or improving key ecosystem functions such as crop yield under a changing environment. © 2014 John Wiley & Sons Ltd.

Spatiotemporal deformation patterns of the Lake Urmia Causeway as characterized by multisensor InSAR analysis.

PubMed

Karimzadeh, Sadra; Matsuoka, Masashi; Ogushi, Fumitaka

2018-04-03

We present deformation patterns in the Lake Urmia Causeway (LUC) in NW Iran based on data collected from four SAR sensors in the form of interferometric synthetic aperture radar (InSAR) time series. Sixty-eight images from Envisat (2004-2008), ALOS-1 (2006-2010), TerraSAR-X (2012-2013) and Sentinel-1 (2015-2017) were acquired, and 227 filtered interferograms were generated using the small baseline subset (SBAS) technique. The rate of line-of-sight (LOS) subsidence of the LUC peaked at 90 mm/year between 2012 and 2013, mainly due to the loss of most of the water in Lake Urmia. Principal component analysis (PCA) was conducted on 200 randomly selected time series of the LUC, and the results are presented in the form of the three major components. The InSAR scores obtained from the PCA were used in a hydro-thermal model to investigate the dynamics of consolidation settlement along the LUC based on detrended water level and temperature data. The results can be used to establish a geodetic network around the LUC to identify more detailed deformation patterns and to help plan future efforts to reduce the possible costs of damage.

Evaluating Brazilian sugarcane expansion effects on soil structure using VESS

USDA-ARS?s Scientific Manuscript database

Increasing global demand for biofuel has accelerated land-use change (LUC) in Brazil, primarily by replacing degraded pasture with sugarcane (Saccharum officinarum). The intensive echanization associated with this LUC has increased concerns regarding soil structural quality (SSQ). Through decades of...

Are We Facing an Epidemic of Self-Injury?

ERIC Educational Resources Information Center

Brumberg, Joan Jacobs

2006-01-01

This article discusses two Cornell research studies on the prevalence of self-injury or "cutting" in today's youth, led by Janis Whitlock, and presents historical perspectives on the issue. The first study randomly surveyed 3,069 undergraduate and graduate students at Cornell University and Princeton University. Data showed that 17 percent of the…

Tough Calls in Tough Times.

ERIC Educational Resources Information Center

Lyman, Linda

1992-01-01

Analyzes school boards' response, in terms of crisis decision making, to a situation in which a superintendent was charged and later convicted of sexually assaulting a 17-year-old female student. Cites "Crucial Decisions," by Irving Janis, and offers recommendations to help board members be more effective decision makers. Presents an outline of…

Rapid obtention of stable, bioluminescent tumor cell lines using a tCD2-luciferase chimeric construct

PubMed Central

2011-01-01

Background Bioluminescent tumor cell lines are experimental tools of major importance for cancer investigation, especially imaging of tumors in xenografted animals. Stable expression of exogenous luciferase in tumor cells combined to systemic injection of luciferin provides an excellent signal/background ratio for external optical imaging. Therefore, there is a need to rationalize and speed up the production of luciferase-positive tumor cell lines representative of multiple tumor phenotypes. For this aim we have designed a fusion gene linking the luciferase 2 protein to the c-terminus of a truncated form of the rat CD2 protein (tCD2-luc2). To allow simultaneous assessment of the wild-type luciferase 2 in a context of tCD2 co-expression, we have made a bicistronic construct for concomitant but separate expression of these two proteins (luc2-IRES-tCD2). Both the mono- and bi-cistronic constructs were transduced in lymphoid and epithelial cells using lentiviral vectors. Results The tCD2-luc2 chimera behaves as a type I membrane protein with surface presentation of CD2 epitopes. One of these epitopes reacts with the OX34, a widely spread, high affinity monoclonal antibody. Stably transfected cells are sorted by flow cytometry on the basis of OX34 staining. In vitro and, moreover, in xenografted tumors, the tCD2-luc2 chimera retains a substantial and stable luciferase activity, although not as high as the wild-type luciferase expressed from the luc2-IRES-tCD2 construct. Expression of the tCD2-luc2 chimera does not harm cell and tumor growth. Conclusion Lentiviral transduction of the chimeric tCD2-luc2 fusion gene allows selection of cell clones with stable luciferase expression in less than seven days without antibiotic selection. We believe that it will be helpful to increase the number of tumor cell lines available for in vivo imaging and assessment of novel therapeutic modalities. On a longer term, the tCD2-luc2 chimera has the potential to be expressed from multi

Large differences in land use emission quantifications implied by definition discrepancies

NASA Astrophysics Data System (ADS)

Stocker, B. D.; Joos, F.

2015-03-01

The quantification of CO2 emissions from anthropogenic land use and land use change (eLUC) is essential to understand the drivers of the atmospheric CO2 increase and to inform climate change mitigation policy. Reported values in synthesis reports are commonly derived from different approaches (observation-driven bookkeeping and process-modelling) but recent work has emphasized that inconsistencies between methods may imply substantial differences in eLUC estimates. However, a consistent quantification is lacking and no concise modelling protocol for the separation of primary and secondary components of eLUC has been established. Here, we review the conceptual differences of eLUC quantification methods and apply an Earth System Model to demonstrate that what is claimed to represent total eLUC differs by up to ~20% when quantified from ESM vs. offline vegetation models. Under a future business-as-usual scenario, differences tend to increase further due to slowing land conversion rates and an increasing impact of altered environmental conditions on land-atmosphere fluxes. We establish how coupled Earth System Models may be applied to separate component fluxes of eLUC arising from the replacement of potential C sinks/sources and the land use feedback and show that secondary fluxes derived from offline vegetation models are conceptually and quantitatively not identical to either, nor their sum. Therefore, we argue that synthesis studies and global carbon budget accountings should resort to the "least common denominator" of different methods, following the bookkeeping approach where only primary land use emissions are quantified under the assumption of constant environmental boundary conditions.

Quantitative measurement of cell membrane receptor internalization by the nanoluciferase reporter: Using the G protein-coupled receptor RXFP3 as a model.

PubMed

Liu, Yu; Song, Ge; Shao, Xiao-Xia; Liu, Ya-Li; Guo, Zhan-Yun

2015-02-01

Nanoluciferase (NanoLuc) is a newly developed small luciferase reporter with the brightest bioluminescence to date. In the present work, we developed NanoLuc as a sensitive bioluminescent reporter to measure quantitatively the internalization of cell membrane receptors, based on the pH dependence of the reporter activity. The G protein-coupled receptor RXFP3, the cognate receptor of relaxin-3/INSL7, was used as a model receptor. We first generated stable HEK293T cells that inducibly coexpressed a C-terminally NanoLuc-tagged human RXFP3 and a C-terminally enhanced green fluorescent protein (EGFP)-tagged human RXFP3. The C-terminal EGFP-tag and NanoLuc-tag had no detrimental effects on the ligand-binding potency and intracellular trafficking of RXFP3. Based on the fluorescence of the tagged EGFP reporter, the ligand-induced RXFP3 internalization was visualized directly under a fluorescence microscope. Based on the bioluminescence of the tagged NanoLuc reporter, the ligand-induced RXFP3 internalization was measured quantitatively by a convenient bioluminescent assay. Coexpression of an EGFP-tagged inactive [E141R]RXFP3 had no detrimental effect on the ligand-binding potency and ligand-induced internalization of the NanoLuc-tagged wild-type RXFP3, suggesting that the mutant RXFP3 and wild-type RXFP3 worked independently. The present bioluminescent internalization assay could be extended to other G protein-coupled receptors and other cell membrane receptors to study ligand-receptor and receptor-receptor interactions. Copyright © 2014 Elsevier B.V. All rights reserved.

Biogeophysical Impacts of Land‐Use Change on Climate Extremes in Low‐Emission Scenarios: Results From HAPPI‐Land

PubMed Central

Seneviratne, Sonia I.; Beyerle, Urs; Boysen, Lena R.; Brovkin, Victor; Davin, Edouard L.; Doelman, Jonathan C.; Kim, Hyungjun; Mitchell, Daniel M.; Nitta, Tomoko; Shiogama, Hideo; Sparrow, Sarah; Stehfest, Elke; van Vuuren, Detlef P.; Wilson, Simon

2018-01-01

Abstract The impacts of land use have been shown to have considerable influence on regional climate. With the recent international commitment to limit global warming to well below 2°C, emission reductions need to be ambitious and could involve major land‐use change (LUC). Land‐based mitigation efforts to curb emissions growth include increasing terrestrial carbon sequestration through reforestation, or the adoption of bioenergy crops. These activities influence local climate through biogeophysical feedbacks, however, it is uncertain how important they are for a 1.5° climate target. This was the motivation for HAPPI‐Land: the half a degree additional warming, prognosis, and projected impacts—land‐use scenario experiment. Using four Earth system models, we present the first multimodel results from HAPPI‐Land and demonstrate the critical role of land use for understanding the characteristics of regional climate extremes in low‐emission scenarios. In particular, our results show that changes in temperature extremes due to LUC are comparable in magnitude to changes arising from half a degree of global warming. We also demonstrate that LUC contributes to more than 20% of the change in temperature extremes for large land areas concentrated over the Northern Hemisphere. However, we also identify sources of uncertainty that influence the multimodel consensus of our results including how LUC is implemented and the corresponding biogeophysical feedbacks that perturb climate. Therefore, our results highlight the urgent need to resolve the challenges in implementing LUC across models to quantify the impacts and consider how LUC contributes to regional changes in extremes associated with sustainable development pathways.

Biogeophysical Impacts of Land-Use Change on Climate Extremes in Low-Emission Scenarios: Results From HAPPI-Land

NASA Astrophysics Data System (ADS)

Hirsch, Annette L.; Guillod, Benoit P.; Seneviratne, Sonia I.; Beyerle, Urs; Boysen, Lena R.; Brovkin, Victor; Davin, Edouard L.; Doelman, Jonathan C.; Kim, Hyungjun; Mitchell, Daniel M.; Nitta, Tomoko; Shiogama, Hideo; Sparrow, Sarah; Stehfest, Elke; van Vuuren, Detlef P.; Wilson, Simon

2018-03-01

The impacts of land use have been shown to have considerable influence on regional climate. With the recent international commitment to limit global warming to well below 2°C, emission reductions need to be ambitious and could involve major land-use change (LUC). Land-based mitigation efforts to curb emissions growth include increasing terrestrial carbon sequestration through reforestation, or the adoption of bioenergy crops. These activities influence local climate through biogeophysical feedbacks, however, it is uncertain how important they are for a 1.5° climate target. This was the motivation for HAPPI-Land: the half a degree additional warming, prognosis, and projected impacts—land-use scenario experiment. Using four Earth system models, we present the first multimodel results from HAPPI-Land and demonstrate the critical role of land use for understanding the characteristics of regional climate extremes in low-emission scenarios. In particular, our results show that changes in temperature extremes due to LUC are comparable in magnitude to changes arising from half a degree of global warming. We also demonstrate that LUC contributes to more than 20% of the change in temperature extremes for large land areas concentrated over the Northern Hemisphere. However, we also identify sources of uncertainty that influence the multimodel consensus of our results including how LUC is implemented and the corresponding biogeophysical feedbacks that perturb climate. Therefore, our results highlight the urgent need to resolve the challenges in implementing LUC across models to quantify the impacts and consider how LUC contributes to regional changes in extremes associated with sustainable development pathways.

Holocene peatland and ice-core data constraints on the timing and magnitude of CO2 emissions from past land use.

PubMed

Stocker, Benjamin David; Yu, Zicheng; Massa, Charly; Joos, Fortunat

2017-02-14

CO 2 emissions from preindustrial land-use change (LUC) are subject to large uncertainties. Although atmospheric CO 2 records suggest only a small land carbon (C) source since 5,000 y before present (5 kyBP), the concurrent C sink by peat buildup could mask large early LUC emissions. Here, we combine updated continuous peat C reconstructions with the land C balance inferred from double deconvolution analyses of atmospheric CO 2 and [Formula: see text]C at different temporal scales to investigate the terrestrial C budget of the Holocene and the last millennium and constrain LUC emissions. LUC emissions are estimated with transient model simulations for diverging published scenarios of LU area change and shifting cultivation. Our results reveal a large terrestrial nonpeatland C source after the Mid-Holocene (66 [Formula: see text] 25 PgC at 7-5 kyBP and 115 [Formula: see text] 27 PgC at 5-3 kyBP). Despite high simulated per-capita CO 2 emissions from LUC in early phases of agricultural development, humans emerge as a driver with dominant global C cycle impacts only in the most recent three millennia. Sole anthropogenic causes for particular variations in the CO 2 record ([Formula: see text]20 ppm rise after 7 kyBP and [Formula: see text]10 ppm fall between 1500 CE and 1600 CE) are not supported. This analysis puts a strong constraint on preindustrial vs. industrial-era LUC emissions and suggests that upper-end scenarios for the extent of agricultural expansion before 1850 CE are not compatible with the C budget thereafter.

Holocene peatland and ice-core data constraints on the timing and magnitude of CO2 emissions from past land use

PubMed Central

Massa, Charly

2017-01-01

CO2 emissions from preindustrial land-use change (LUC) are subject to large uncertainties. Although atmospheric CO2 records suggest only a small land carbon (C) source since 5,000 y before present (5 kyBP), the concurrent C sink by peat buildup could mask large early LUC emissions. Here, we combine updated continuous peat C reconstructions with the land C balance inferred from double deconvolution analyses of atmospheric CO2 and δ13C at different temporal scales to investigate the terrestrial C budget of the Holocene and the last millennium and constrain LUC emissions. LUC emissions are estimated with transient model simulations for diverging published scenarios of LU area change and shifting cultivation. Our results reveal a large terrestrial nonpeatland C source after the Mid-Holocene (66 ± 25 PgC at 7–5 kyBP and 115 ± 27 PgC at 5–3 kyBP). Despite high simulated per-capita CO2 emissions from LUC in early phases of agricultural development, humans emerge as a driver with dominant global C cycle impacts only in the most recent three millennia. Sole anthropogenic causes for particular variations in the CO2 record (∼20 ppm rise after 7 kyBP and ∼10 ppm fall between 1500 CE and 1600 CE) are not supported. This analysis puts a strong constraint on preindustrial vs. industrial-era LUC emissions and suggests that upper-end scenarios for the extent of agricultural expansion before 1850 CE are not compatible with the C budget thereafter. PMID:28137849

Soil physical quality response to sugarcane expansion in Brazil

USDA-ARS?s Scientific Manuscript database

With an increasing global demand for bioenergy, the rate of land-use change (LUC) is increasing rapidly to support cellulosic feedstock production. In Brazil, sugarcane (Saccharum officinarum) expansion to produce ethanol is displacing degraded pastures, but long-term effects of this LUC are unknown...

Early Student Support for the Study of Inertial Motions in the Arctic Ocean

DTIC Science & Technology

2015-09-30

Dosser, Hayley V., Sasan J. Ghaemsaidi, Thomas Peacock , and Luc Rainville, x. Internal Wave 5 Propagation and Stability in the Western Arctic...Ocean. In preparation for J. Phys. Oceanogr. Ghaemsaidi, Sasan J., Hayley V. Dosser, Luc Rainville, and Thomas Peacock , 2015. The impact of multiple

Soil quality indexing strategies for evaluating sugarcane expansion in Brazil

USDA-ARS?s Scientific Manuscript database

Increasing demands for biofuels have intensified the land use change (LUC) for sugarcane cropping expansion in Brazil. Assessments of LUC-induced changes on soil quality (SQ) are essential for quantifying and monitoring the sustainability of sugarcane production over time. Since there is not a unive...

In vivo imaging of an inducible oncogenic tumor antigen visualizes tumor progression and predicts CTL tolerance.

PubMed

Buschow, Christian; Charo, Jehad; Anders, Kathleen; Loddenkemper, Christoph; Jukica, Ana; Alsamah, Wisam; Perez, Cynthia; Willimsky, Gerald; Blankenstein, Thomas

2010-03-15

Visualizing oncogene/tumor Ag expression by noninvasive imaging is of great interest for understanding processes of tumor development and therapy. We established transgenic (Tg) mice conditionally expressing a fusion protein of the SV40 large T Ag and luciferase (TagLuc) that allows monitoring of oncogene/tumor Ag expression by bioluminescent imaging upon Cre recombinase-mediated activation. Independent of Cre-mediated recombination, the TagLuc gene was expressed at low levels in different tissues, probably due to the leakiness of the stop cassette. The level of spontaneous TagLuc expression, detected by bioluminescent imaging, varied between the different Tg lines, depended on the nature of the Tg expression cassette, and correlated with Tag-specific CTL tolerance. Following liver-specific Cre-loxP site-mediated excision of the stop cassette that separated the promoter from the TagLuc fusion gene, hepatocellular carcinoma development was visualized. The ubiquitous low level TagLuc expression caused the failure of transferred effector T cells to reject Tag-expressing tumors rather than causing graft-versus-host disease. This model may be useful to study different levels of tolerance, monitor tumor development at an early stage, and rapidly visualize the efficacy of therapeutic intervention versus potential side effects of low-level Ag expression in normal tissues.

Spatial model of land use change related to sediment yield (case study: Cipeles and Cilutung watershed, West Java)

NASA Astrophysics Data System (ADS)

Wulandari, D. W.; Kusratmoko, E.; Indra, T. L.

2018-05-01

Land use changes (LUC) as a result of increasing human need for space are likely to destroy the hydrological function of the watershed, increase land degradation, stimulate erosion and drive the process of sedimentation. This study aimed to predict LUC during the period 1990 to 2030 in relation to sediment yield in Cilutung and Cipeles Watershed, West Java. LUC were simulated following the model of Cellular Automata-Marcov Chain, whereas land use composition in 2030 was predicted using Land Change Modeler on Idrisi Selva Software. Elevation, slope, distance from road, distance from river, and distance from settlement were selected as driving factors for LUC in this study. Erosion and sediment yield were predicted using WATEM/SEDEM model based on land use, rainfall, soil texture and topography. The results showed that the areas of forest and shrub have slightly declined up to 5% during the period 1990 to 2016, generally being converted into rice fields, settlements, non-irrigated fields and plantations. In addition, rice fields, settlements, and plantations were expected to substantially increase up to 50% in 2030. Furthermore, the study also revealed that erosion and sediment yield tend to increase every year. This is likely associated with LUC occurring in Cipeles and Cilutung Watershed.

Uncertainties in the land-use flux resulting from land-use change reconstructions and gross land transitions

NASA Astrophysics Data System (ADS)

Bayer, Anita D.; Lindeskog, Mats; Pugh, Thomas A. M.; Anthoni, Peter M.; Fuchs, Richard; Arneth, Almut

2017-02-01

Land-use and land-cover (LUC) changes are a key uncertainty when attributing changes in measured atmospheric CO2 concentration to its sinks and sources and must also be much better understood to determine the possibilities for land-based climate change mitigation, especially in the light of human demand on other land-based resources. On the spatial scale typically used in terrestrial ecosystem models (0.5 or 1°) changes in LUC over time periods of a few years or more can include bidirectional changes on the sub-grid level, such as the parallel expansion and abandonment of agricultural land (e.g. in shifting cultivation) or cropland-grassland conversion (and vice versa). These complex changes between classes within a grid cell have often been neglected in previous studies, and only net changes of land between natural vegetation cover, cropland and pastures accounted for, mainly because of a lack of reliable high-resolution historical information on gross land transitions, in combination with technical limitations within the models themselves. In the present study we applied a state-of-the-art dynamic global vegetation model with a detailed representation of croplands and carbon-nitrogen dynamics to quantify the uncertainty in terrestrial ecosystem carbon stocks and fluxes arising from the choice between net and gross representations of LUC. We used three frequently applied global, one recent global and one recent European LUC datasets, two of which resolve gross land transitions, either in Europe or in certain tropical regions. When considering only net changes, land-use-transition uncertainties (expressed as 1 standard deviation around decadal means of four models) in global carbon emissions from LUC (ELUC) are ±0.19, ±0.66 and ±0.47 Pg C a-1 in the 1980s, 1990s and 2000s, respectively, or between 14 and 39 % of mean ELUC. Carbon stocks at the end of the 20th century vary by ±11 Pg C for vegetation and ±37 Pg C for soil C due to the choice of LUC

Development of New Low Temperature Systems at Janis Research

NASA Astrophysics Data System (ADS)

Shvarts, V.; Jirmanus, M. N.; Zhao, Z.

2006-09-01

A new line of ultra low loss He-3 and He-4 superconducting magnet systems and UHV compatible He-3 systems for scanning tunneling microscopy studies have been developed during the past year. This paper discusses the details of the design and performance for the various systems.

Longitudinal Imaging of Cancer Cell Metastases in Two Preclinical Models: A Correlation of Noninvasive Imaging to Histopathology

PubMed Central

Adiseshaiah, Pavan P.; Patel, Nimit L.; Ileva, Lilia V.; Kalen, Joseph D.; Haines, Diana C.; McNeil, Scott E.

2014-01-01

Metastatic spread is the leading cause of death from cancer. Early detection of cancer at primary and metastatic sites by noninvasive imaging modalities would be beneficial for both therapeutic intervention and disease management. Noninvasive imaging modalities such as bioluminescence (optical), positron emission tomography (PET)/X-ray computed tomography (CT), and magnetic resonance imaging (MRI) can provide complementary information and accurately measure tumor growth as confirmed by histopathology. Methods. We validated two metastatic tumor models, MDA-MD-231-Luc and B16-F10-Luc intravenously injected, and 4T1-Luc cells orthotopically implanted into the mammary fat pad. Longitudinal whole body bioluminescence imaging (BLI) evaluated metastasis, and tumor burden of the melanoma cell line (B16-F10-Luc) was correlated with (PET)/CT and MRI. In addition, ex vivo imaging evaluated metastasis in relevant organs and histopathological analysis was used to confirm imaging. Results. BLI revealed successful colonization of cancer cells in both metastatic tumor models over a 4-week period. Furthermore, lung metastasis of B16-F10-Luc cells imaged by PET/CT at week four showed a strong correlation (R 2 = 0.9) with histopathology. The presence and degree of metastasis as determined by imaging correlated (R 2 = 0.7) well with histopathology findings. Conclusions. We validated two metastatic tumor models by longitudinal noninvasive imaging with good histopathology correlation. PMID:24724022

Validation of the use of an artificial mitochondrial reporter DNA vector containing a Cytomegalovirus promoter for mitochondrial transgene expression.

PubMed

Yamada, Yuma; Ishikawa, Takuya; Harashima, Hideyoshi

2017-08-01

Mitochondria have their own gene expression system that is independent of the nuclear system, and control cellular functions in cooperation with the nucleus. While a number of useful technologies for achieving nuclear transgene expression have been reported, only a few have focused on mitochondria. In this study, we validated the utility of an artificial mitochondrial DNA vector with a virus promoter on mitochondrial transgene expression. We designed and constructed pCMV-mtLuc (CGG) that contains a CMV promotor derived from Cytomegalovirus and an artificial mitochondrial genome with a NanoLuc (Nluc) luciferase gene that records adjustments to the mitochondrial codon system. Nluc luciferase activity measurements showed that the pCMV-mtLuc (CGG) efficiently produced the Nluc luciferase protein in human HeLa cells. Moreover, we optimized the mitochondrial transfection of pCMV-mtLuc (CGG) using a MITO-Porter system, a liposome-based carrier for mitochondrial delivery via membrane fusion. As a result, we found that transfection of pCMV-mtLuc (CGG) by MITO-Porter modified with the KALA peptide (cationic amphipathic cell-penetrating peptide) showed a high mitochondrial transgene expression. The developed mitochondrial transgene expression system represents a potentially useful tool for the fields of nanoscience and nanotechnology for controlling the intracellular microenvironment via the regulation of mitochondrial function and promises to open additional innovative research fields of study. Copyright © 2017 Elsevier Ltd. All rights reserved.

Coping with pregnancy resolution among never-married women.

PubMed

Bracken, Michael B; Klerman, Lorraine V; Bracken, Maryann

1978-04-01

The Janis-Mann model of decision-making provides the theoretical orientation for empirical analyses of decisions to deliver or abort in matched samples of never-married women. Results focus on four variables: happiness about pregnancy; initial acceptance of delivery or abortion; ease of decision-making; and satisfaction with final choice. Path analyses summarize findings, which are discussed in terms of conflict resolution strategies.

Influence of Culture and Personality on Determinants of Cognitive Processes Under Conditions of Uncertainty

DTIC Science & Technology

2004-05-14

Tal , Y., Raviv , A., & Spitzer, A., 1999). Janis and Mann (1977) suggested that situational conditions determine how individuals cope with decision...and ignore contrary information relative to non-stressful conditions, which can have disastrous consequences. Bar- Tal , Raviv , and Spitzer (1999...1176. Bar- Tal , Y., Raviv , A., & Spitzer, A. (1999). The need and ability to achieve cognitive structuring: Individual differences that moderate

Representing anthropogenic gross land use change, wood harvest, and forest age dynamics in a global vegetation model ORCHIDEE-MICT v8.4.2

NASA Astrophysics Data System (ADS)

Yue, Chao; Ciais, Philippe; Luyssaert, Sebastiaan; Li, Wei; McGrath, Matthew J.; Chang, Jinfeng; Peng, Shushi

2018-01-01

Land use change (LUC) is among the main anthropogenic disturbances in the global carbon cycle. Here we present the model developments in a global dynamic vegetation model ORCHIDEE-MICT v8.4.2 for a more realistic representation of LUC processes. First, we included gross land use change (primarily shifting cultivation) and forest wood harvest in addition to net land use change. Second, we included sub-grid evenly aged land cohorts to represent secondary forests and to keep track of the transient stage of agricultural lands since LUC. Combination of these two features allows the simulation of shifting cultivation with a rotation length involving mainly secondary forests instead of primary ones. Furthermore, a set of decision rules regarding the land cohorts to be targeted in different LUC processes have been implemented. Idealized site-scale simulation has been performed for miombo woodlands in southern Africa assuming an annual land turnover rate of 5 % grid cell area between forest and cropland. The result shows that the model can correctly represent forest recovery and cohort aging arising from agricultural abandonment. Such a land turnover process, even though without a net change in land cover, yields carbon emissions largely due to the imbalance between the fast release from forest clearing and the slow uptake from agricultural abandonment. The simulation with sub-grid land cohorts gives lower emissions than without, mainly because the cleared secondary forests have a lower biomass carbon stock than the mature forests that are otherwise cleared when sub-grid land cohorts are not considered. Over the region of southern Africa, the model is able to account for changes in different forest cohort areas along with the historical changes in different LUC activities, including regrowth of old forests when LUC area decreases. Our developments provide possibilities to account for continental or global forest demographic change resulting from past anthropogenic and

[Establishment of an iRFP and luciferase dual-color fluorescence-traced hepatocellular carcinoma transplantation model in nude mice].

PubMed

Li, Hongjun; Yang, Tianhua; Huang, Yanping; Liu, Mingzhu; Qin, Zhongqiang; Chu, Fei; Li, Zhenghong; Li, Yonghai

2017-11-01

Objective To establish a hepatocellular carcinoma xenograft model in nude mice which could stably express gene and be monitored dynamically. Methods We first constructed the lentiviral particles containing luciferase (Luc) and near-infrared fluorescent protein (iRFP) and puromycin resistance gene, and then transduced them into the HepG2 hepatoma cells. The cell line stably expressing Luc and iRFP genes were screened and inoculated into nude mice to establish xenograft tumor model. Tumor growth was monitored using in vivo imaging system. HE staining and immunohistochemistry were used to evaluate the pathological features and tumorigenic ability. Results HepG2 cells stably expressing iRFP and Luc were obtained; with the engineered cell line, xenograft model was successfully established with the features of proper tumor developing time and high rate of tumor formation as well as typical pathological features as showed by HE staining and immunohistochemistry. Conclusion Hepatocellular carcinoma model in nude mice with the features of stable gene expression and dynamical monitoring has been established successfully with the HepG2-iRFP-Luc cell line.

Urban Land Use Change Effects on Below and Aboveground Carbon Stocks—a Global Perspective and Future Research Needs

NASA Astrophysics Data System (ADS)

Pouyat, R. V.; Chen, Y.; Yesilonis, I.; Day, S.

2014-12-01

Land use change (LUC) has a significant impact on both above- and below-ground carbon (C) stocks; however, little is known about the net effects of urban LUC on the C cycle and climate system. Moreover, as climate change becomes an increasingly pressing concern, there is growing evidence that urban policy and management decisions can have significant regional impacts on C dynamics. Soil organic carbon (SOC) varies significantly across ecoregions at global and continental scales due to differential sensitivity of primary production, substrate quality, and organic matter decay to changes in temperature and soil moisture. These factors are highly modified by urban LUC due to vegetation removal, soil relocation and disruption, pollution, urban heat island effects, and increased atmospheric CO2 concentrations. As a result, on a global scale urban LUC differentially affects the C cycle from ecoregion to ecoregion. For urban ecosystems, the data collected thus far suggests urbanization can lead to both an increase and decrease in soil C pools and fluxes, depending on the native ecosystem being impacted by urban development. For example, in drier climates, urban landscapes accumulate higher C densities than the native ecosystems they replaced. Results suggest also that soil C storage in urban ecosystems is highly variable with very high (> 20.0) and low (< 2.0) C densities (kg m-2 to a 1 m depth) present in the landscape at any one time. Moreover, similar to non-urban soils, total SOC densities are consistently 2-fold greater than aboveground stocks. For those soils with low SOC densities, there is potential to increase C sequestration through management, but specific urban related management practices need to be evaluated. In addition, urban LUC is a human-driven process and thus can be modified or adjusted to reduce its impacts on the C cycle. For example, policies that influence development patterns, population density, management practices, and other human factors can

IA Channels Encoded by Kv1.4 and Kv4.2 Regulate Circadian Period of PER2 Expression in the Suprachiasmatic Nucleus.

PubMed

Granados-Fuentes, Daniel; Hermanstyne, Tracey O; Carrasquillo, Yarimar; Nerbonne, Jeanne M; Herzog, Erik D

2015-10-01

Neurons in the suprachiasmatic nucleus (SCN), the master circadian pacemaker in mammals, display daily rhythms in electrical activity with more depolarized resting potentials and higher firing rates during the day than at night. Although these daily variations in the electrical properties of SCN neurons are required for circadian rhythms in physiology and behavior, the mechanisms linking changes in neuronal excitability to the molecular clock are not known. Recently, we reported that mice deficient for either Kcna4 (Kv1.4(-/-)) or Kcnd2 (Kv4.2(-/-); but not Kcnd3, Kv4.3(-/-)), voltage-gated K(+) (Kv) channel pore-forming subunits that encode subthreshold, rapidly activating, and inactivating K(+) currents (IA), have shortened (0.5 h) circadian periods in SCN firing and in locomotor activity compared with wild-type (WT) mice. In the experiments here, we used a mouse (Per2(Luc)) line engineered with a bioluminescent reporter construct, PERIOD2::LUCIFERASE (PER2::LUC), replacing the endogenous Per2 locus, to test the hypothesis that the loss of Kv1.4- or Kv4.2-encoded IA channels also modifies circadian rhythms in the expression of the clock protein PERIOD2 (PER2). We found that SCN explants from Kv1.4(-/-)Per2(Luc) and Kv4.2(-/-) Per2(Luc), but not Kv4.3(-/-)Per2(Luc), mice have significantly shorter (by approximately 0.5 h) circadian periods in PER2 rhythms, compared with explants from Per2(Luc) mice, revealing that the membrane properties of SCN neurons feedback to regulate clock (PER2) expression. The combined loss of both Kv1.4- and Kv4.2-encoded IA channels in Kv1.4(-/-)/Kv4.2(-/-)Per2(Luc) SCN explants did not result in any further alterations in PER2 rhythms. Interestingly, however, mice lacking both Kv1.4 and Kv4.2 show a striking (approximately 1.8 h) advance in their daily activity onset in a light cycle compared with WT mice, suggesting additional roles for Kv1.4- and Kv4.2-encoded IA channels in controlling the light-dependent responses of neurons within

Whole-genome re-sequencing of two Italian tomato landraces reveals sequence variations in genes associated with stress tolerance, fruit quality and long shelf-life traits.

PubMed

Tranchida-Lombardo, Valentina; Aiese Cigliano, Riccardo; Anzar, Irantzu; Landi, Simone; Palombieri, Samuela; Colantuono, Chiara; Bostan, Hamed; Termolino, Pasquale; Aversano, Riccardo; Batelli, Giorgia; Cammareri, Maria; Carputo, Domenico; Chiusano, Maria Luisa; Conicella, Clara; Consiglio, Federica; D'Agostino, Nunzio; De Palma, Monica; Di Matteo, Antonio; Grandillo, Silvana; Sanseverino, Walter; Tucci, Marina; Grillo, Stefania

2017-11-14

Tomato is a high value crop and the primary model for fleshy fruit development and ripening. Breeding priorities include increased fruit quality, shelf life and tolerance to stresses. To contribute towards this goal, we re-sequenced the genomes of Corbarino (COR) and Lucariello (LUC) landraces, which both possess the traits of plant adaptation to water deficit, prolonged fruit shelf-life and good fruit quality. Through the newly developed pipeline Reconstructor, we generated the genome sequences of COR and LUC using datasets of 65.8 M and 56.4 M of 30-150 bp paired-end reads, respectively. New contigs including reads that could not be mapped to the tomato reference genome were assembled, and a total of 43, 054 and 44, 579 gene loci were annotated in COR and LUC. Both genomes showed novel regions with similarity to Solanum pimpinellifolium and Solanum pennellii. In addition to small deletions and insertions, 2, 000 and 1, 700 single nucleotide polymorphisms (SNPs) could exert potentially disruptive effects on 1, 371 and 1, 201 genes in COR and LUC, respectively. A detailed survey of the SNPs occurring in fruit quality, shelf life and stress tolerance related-genes identified several candidates of potential relevance. Variations in ethylene response components may concur in determining peculiar phenotypes of COR and LUC. © The Author 2017. Published by Oxford University Press on behalf of Kazusa DNA Research Institute.

Land-Use Change and the Billion Ton 2016 Resource Assessment: Understanding the Effects of Land Management on Environmental Indicators

NASA Astrophysics Data System (ADS)

Kline, K. L.; Eaton, L. M.; Efroymson, R.; Davis, M. R.; Dunn, J.; Langholtz, M. H.

2016-12-01

The federal government, led by the U.S. Department of Energy (DOE), quantified potential U.S. biomass resources for expanded production of renewable energy and bioproducts in the 2016 Billion-Ton Report: Advancing Domestic Resources for a Thriving Bioeconomy (BT16) (DOE 2016). Volume 1 of the report provides analysis of projected supplies from 2015 to2040. Volume 2 (forthcoming) evaluates changes in environmental indicators for water quality and quantity, carbon, air quality, and biodiversity associated with production scenarios in BT16 volume 1. This presentation will review land-use allocations under the projected biomass production scenarios and the changes in land management that are implied, including drivers of direct and indirect LUC. National and global concerns such as deforestation and displacement of food production are addressed. The choice of reference scenario, input parameters and constraints (e.g., regarding land classes, availability, and productivity) drive LUC results in any model simulation and are reviewed to put BT16 impacts into context. The principal LUC implied in BT16 supply scenarios involves the transition of 25-to-47 million acres (net) from annual crops in 2015 baseline to perennial cover by 2040 under the base case and 3% yield growth case, respectively. We conclude that clear definitions of land parameters and effects are essential to assess LUC. A lack of consistency in parameters and outcomes of historic LUC analysis in the U.S. underscores the need for science-based approaches.

Beta emitters rhenium-188 and lutetium-177 are equally effective in radioimmunotherapy of HPV-positive experimental cervical cancer.

PubMed

Phaeton, Rebecca; Jiang, Zewei; Revskaya, Ekaterina; Fisher, Darrell R; Goldberg, Gary L; Dadachova, Ekaterina

2016-01-01

Cervical cancer caused by the infection with the human papillomavirus (HPV) remains the fourth leading killer of women worldwide. Therefore, more efficacious treatments are needed. We are developing radioimmunotherapy (RIT) of HPV-positive cervical cancers by targeting E6 and E7 viral oncoproteins expressed by the cancer cells with the radiolabeled monoclonal antibodies (mAbs). To investigate the influence of different radionuclides on the RIT efficacy-we performed RIT of experimental cervical cancer with Rhenium-188 ((188) Re) and Lutetium-177 ((177) Lu)-labeled mAb C1P5 to E6. The biodistribution of (188) Re- and (177) Lu-labeled C1P5 was performed in nude female mice bearing CasKi cervical cancer xenografts and the radiation dosimetry calculations for the tumors and organs were carried out. For RIT the mice were treated with 7.4 MBq of either (188) Re-C1P5 or (177) Lu-C1P5 or left untreated, and observed for their tumor size for 28 days. The levels of (188) Re- and (177) Lu-C1P5 mAbs-induced double-strand breaks in CasKi tumors were compared on days 5 and 10 post treatment by staining with anti-gamma H2AX antibody. The radiation doses to the heart and lungs were similar for both (177) Lu-C1P5 and (188) Re-C1P5. The dose to the liver was five times higher for (177) Lu-C1P5. The doses to the tumor were 259 and 181 cGy for (177) Lu-C1P5 and (188) Re-C1P5, respectively. RIT with either (177) Lu-C1P5 or (188) Re-C1P5 was equally effective in inhibiting tumor growth when each was compared to the untreated controls (P = 0.001). On day 5 there was a pronounced staining for gamma H2AX foci in (177) Lu-C1P5 group only and on day 10 it was observed in both (177) Lu-C1P5 and (188) Re-C1P5 groups. (188) Re- and (177) Lu-labeled mAbs were equally effective in arresting the growth of CasKi cervical tumors. Thus, both of these radionuclides are candidates for the clinical trials of this approach in patients with advanced, recurrent or metastatic cervical cancer. © 2015 The

Army Synthetic Validity Project Report of Phase 2 Results. Volume 2. Appendixes

DTIC Science & Technology

1990-10-01

to Equipment & Food o Personal Hygine - Field & Garrison (4) o Kitchen Equipment - Garrison o Field Preparation of Foods & Equipment o Food, Field...Results: Volume II: Appendi i 12. PERSONAL AUTHOR(S) Wise, Lauress L. (AIR); Peterson, Norman G.; Houston, Janis (PDRI); Hoffman, R. Gene Campbell, John...o Handling KIA o Personal Hygiene & Preventive Medicine Numbers in parentheses indicate the number of participants that identified the task as

Potent Effects of Flavonoid Nobiletin on Amplitude, Period, and Phase of the Circadian Clock Rhythm in PER2::LUCIFERASE Mouse Embryonic Fibroblasts.

PubMed

Shinozaki, Ayako; Misawa, Kenichiro; Ikeda, Yuko; Haraguchi, Atsushi; Kamagata, Mayo; Tahara, Yu; Shibata, Shigenobu

2017-01-01

Flavonoids are natural polyphenols that are widely found in plants. The effects of flavonoids on obesity and numerous diseases such as cancer, diabetes, and Alzheimer's have been well studied. However, little is known about the relationships between flavonoids and the circadian clock. In this study, we show that continuous or transient application of flavonoids to the culture medium of embryonic fibroblasts from PER2::LUCIFERASE (PER2::LUC) mice induced various modifications in the circadian clock amplitude, period, and phase. Transient application of some of the tested flavonoids to cultured cells induced a phase delay of the PER2::LUC rhythm at the down slope phase. In addition, continuous application of the polymethoxy flavonoids nobiletin and tangeretin increased the amplitude and lengthened the period of the PER2::LUC rhythm. The nobiletin-induced phase delay was blocked by co-treatment with U0126, an ERK inhibitor. In summary, among the tested flavonoids, polymethoxy flavones increased the amplitude, lengthened the period, and delayed the phase of the PER2::LUC circadian rhythm. Therefore, foods that contain polymethoxy flavones may have beneficial effects on circadian rhythm disorders and jet lag.

Transnasal endoscopic medial maxillectomy in recurrent maxillary sinus inverted papilloma.

PubMed

Kamel, Reda H; Abdel Fattah, Ahmed F; Awad, Ayman G

2014-12-01

Maxillary sinus inverted papilloma entails medial maxillectomy and is associated with high incidence of recurrence. To study the impact of prior surgery on recurrence rate after transnasal endoscopic medial maxillectomy. Eighteen patients with primary and 33 with recurrent maxillary sinus inverted papilloma underwent transnasal endoscopic medial maxillectomy. Caldwell-Luc operation was the primary surgery in 12 patients, transnasal endoscopic resection in 20, and midfacial degloving technique in one. The follow-up period ranged between 2 to 19.5 years with an average of 8.8 years. Recurrence was detected in 8/51 maxillary sinus inverted papilloma patients (15.7 %), 1/18 of primary cases (5.5 %), 7/33 of recurrent cases (21.2 %); 3/20 of the transnasal endoscopic resection group (15%) and 4/12 of the Caldwell-Luc group (33.3%). Redo transnasal endoscopic medial maxillectomy was followed by a single recurrence in the Caldwell-Luc group (25%), and no recurrence in the other groups. Recurrence is more common in recurrent maxillary sinus inverted papilloma than primary lesions. Recurrent maxillary sinus inverted papilloma after Caldwell-Luc operation has higher incidence of recurrence than after transnasal endoscopic resection.

Quantifying differences in land use emission estimates implied by definition discrepancies

NASA Astrophysics Data System (ADS)

Stocker, B. D.; Joos, F.

2015-11-01

The quantification of CO2 emissions from anthropogenic land use and land use change (eLUC) is essential to understand the drivers of the atmospheric CO2 increase and to inform climate change mitigation policy. Reported values in synthesis reports are commonly derived from different approaches (observation-driven bookkeeping and process-modelling) but recent work has emphasized that inconsistencies between methods may imply substantial differences in eLUC estimates. However, a consistent quantification is lacking and no concise modelling protocol for the separation of primary and secondary components of eLUC has been established. Here, we review differences of eLUC quantification methods and apply an Earth System Model (ESM) of Intermediate Complexity to quantify them. We find that the magnitude of effects due to merely conceptual differences between ESM and offline vegetation model-based quantifications is ~ 20 % for today. Under a future business-as-usual scenario, differences tend to increase further due to slowing land conversion rates and an increasing impact of altered environmental conditions on land-atmosphere fluxes. We establish how coupled Earth System Models may be applied to separate secondary component fluxes of eLUC arising from the replacement of potential C sinks/sources and the land use feedback and show that secondary fluxes derived from offline vegetation models are conceptually and quantitatively not identical to either, nor their sum. Therefore, we argue that synthesis studies should resort to the "least common denominator" of different methods, following the bookkeeping approach where only primary land use emissions are quantified under the assumption of constant environmental boundary conditions.

Regionalization of land-use impacts on streamflow using a network of paired catchments

NASA Astrophysics Data System (ADS)

Ochoa-Tocachi, Boris F.; Buytaert, Wouter; De Bièvre, Bert

2016-09-01

Quantifying the impact of land use and cover (LUC) change on catchment hydrological response is essential for land-use planning and management. Yet hydrologists are often not able to present consistent and reliable evidence to support such decision-making. The issue tends to be twofold: a scarcity of relevant observations, and the difficulty of regionalizing any existing observations. This study explores the potential of a paired catchment monitoring network to provide statistically robust, regionalized predictions of LUC change impact in an environment of high hydrological variability. We test the importance of LUC variables to explain hydrological responses and to improve regionalized predictions using 24 catchments distributed along the Tropical Andes. For this, we calculate first 50 physical catchment properties, and then select a subset based on correlation analysis. The reduced set is subsequently used to regionalize a selection of hydrological indices using multiple linear regression. Contrary to earlier studies, we find that incorporating LUC variables in the regional model structures increases significantly regression performance and predictive capacity for 66% of the indices. For the runoff ratio, baseflow index, and slope of the flow duration curve, the mean absolute error reduces by 53% and the variance of the residuals by 79%, on average. We attribute the explanatory capacity of LUC in the regional model to the pairwise monitoring setup, which increases the contrast of the land-use signal in the data set. As such, it may be a useful strategy to optimize data collection to support watershed management practices and improve decision-making in data-scarce regions.

Increase in DNA vaccine efficacy by virosome delivery and co-expression of a cytolytic protein.

PubMed

Gargett, Tessa; Grubor-Bauk, Branka; Miller, Darren; Garrod, Tamsin; Yu, Stanley; Wesselingh, Steve; Suhrbier, Andreas; Gowans, Eric J

2014-06-01

The potential of DNA vaccines has not been realised due to suboptimal delivery, poor antigen expression and the lack of localised inflammation, essential for antigen presentation and an effective immune response to the immunogen. Initially, we examined the delivery of a DNA vaccine encoding a model antigen, luciferase (LUC), to the respiratory tract of mice by encapsulation in a virosome. Virosomes that incorporated influenza virus haemagglutinin effectively delivered DNA to cells in the mouse respiratory tract and resulted in antigen expression and systemic and mucosal immune responses to the immunogen after an intranasal (IN) prime/intradermal (ID) boost regimen, whereas a multidose ID regimen only generated systemic immunity. We also examined systemic immune responses to LUC after ID vaccination with a DNA vaccine, which also encoded one of the several cytolytic or toxic proteins. Although the herpes simplex virus thymidine kinase, in the presence of the prodrug, ganciclovir, resulted in cell death, this failed to increase the humoral or cell-mediated immune responses. In contrast, the co-expression of LUC with the rotavirus non-structural protein 4 (NSP4) protein or a mutant form of mouse perforin, proteins which are directly cytolytic, resulted in increased LUC-specific humoral and cell-mediated immunity. On the other hand, co-expression of LUC with diphtheria toxin subunit A or overexpression of perforin or NSP4 resulted in a lower level of immunity. In summary, the efficacy of DNA vaccines can be improved by targeted IN delivery of DNA or by the induction of cell death in vaccine-targeted cells after ID delivery.

Development of a recombinant human ovarian (BG1) cell line containing estrogen receptor α and β for improved detection of estrogenic/antiestrogenic chemicals

PubMed Central

Brennan, Jennifer C.; Bassal, Arzoo; He, Guochun; Denison, Michael S.

2016-01-01

Estrogenic endocrine disrupting chemicals are found in environmental and biological samples, commercial and consumer products, food, and numerous other sources. Given their ubiquitous nature and potential for adverse effects, there is a critical need for rapidly detecting these chemicals. We developed an estrogen-responsive recombinant human ovarian (BG1Luc4E2) cell line recently accepted by the USEPA and OECD as a bioanalytical method to detect estrogen receptor (ER) agonists/antagonists. Unfortunately, these cells appear to contain only one of the two known ER isoforms, ERα but not ERβ, and the differential ligand selectivity of these ERs indicates that the currently accepted screening method only detects a subset of total estrogenic chemicals. To improve the estrogen screening bioassay, BG1Luc4E2 cells were stably transfected with an ERβ expression plasmid and positive clones identified using ERβ-selective ligands (genistein and Br-ERβ-041). A highly responsive clone (BG1LucERβc9) was identified that exhibited greater sensitivity and responsiveness to ERβ-selective ligands than BG1Luc4E2 cells and qRT-PCR confirmed the presence of ERβ expression in these cells. Screening of pesticides and industrial chemicals identified chemicals that preferentially stimulated ERβ-dependent reporter gene expression. Together, these results not only demonstrate the utility of this dual ER recombinant cell line for detecting a broader range of estrogenic chemicals than the current BG1Luc4E2 cell line, but screening with both cell lines allows identification of ERα and ERβ-selective chemicals. PMID:26139245

A causal analysis framework for land-use change and the potential role of bioenergy policy

DOE PAGES

Efroymson, Rebecca A.; Kline, Keith L.; Angelsen, Arild; ...

2016-10-05

Here we propose a causal analysis framework to increase the reliability of land-use change (LUC) models and the accuracy of net greenhouse gas (GHG) emissions calculations for biofuels. The health-sciences-inspired framework is used here to determine probable causes of LUC, with an emphasis on bioenergy and deforestation. Calculations of net GHG emissions for LUC are critical in determining whether a fuel qualifies as a biofuel or advanced biofuel category under national (U.S., U.K.), state (California), and European Union regulations. Biofuel policymakers and scientists continue to discuss whether presumed indirect land-use change (ILUC) estimates, which often involve deforestation, should be includedmore » in GHG accounting for biofuel pathways. Current estimates of ILUC for bioenergy rely largely on economic simulation models that focus on causal pathways involving global commodity trade and use coarse land cover data with simple land classification systems. ILUC estimates are highly uncertain, partly because changes are not clearly defined and key causal links are not sufficiently included in the models. The proposed causal analysis framework begins with a definition of the change that has occurred and proceeds to a strength-of-evidence approach based on types of epidemiological evidence including plausibility of the relationship, completeness of the causal pathway, spatial co-occurrence, time order, analogous agents, simulation model results, and quantitative agent response relationships.Lastly, we discuss how LUC may be allocated among probable causes for policy purposes and how the application of the framework has the potential to increase the validity of LUC models and resolve ILUC and biofuel controversies.« less

A causal analysis framework for land-use change and the potential role of bioenergy policy

DOE Office of Scientific and Technical Information (OSTI.GOV)

Efroymson, Rebecca A.; Kline, Keith L.; Angelsen, Arild

Here we propose a causal analysis framework to increase the reliability of land-use change (LUC) models and the accuracy of net greenhouse gas (GHG) emissions calculations for biofuels. The health-sciences-inspired framework is used here to determine probable causes of LUC, with an emphasis on bioenergy and deforestation. Calculations of net GHG emissions for LUC are critical in determining whether a fuel qualifies as a biofuel or advanced biofuel category under national (U.S., U.K.), state (California), and European Union regulations. Biofuel policymakers and scientists continue to discuss whether presumed indirect land-use change (ILUC) estimates, which often involve deforestation, should be includedmore » in GHG accounting for biofuel pathways. Current estimates of ILUC for bioenergy rely largely on economic simulation models that focus on causal pathways involving global commodity trade and use coarse land cover data with simple land classification systems. ILUC estimates are highly uncertain, partly because changes are not clearly defined and key causal links are not sufficiently included in the models. The proposed causal analysis framework begins with a definition of the change that has occurred and proceeds to a strength-of-evidence approach based on types of epidemiological evidence including plausibility of the relationship, completeness of the causal pathway, spatial co-occurrence, time order, analogous agents, simulation model results, and quantitative agent response relationships.Lastly, we discuss how LUC may be allocated among probable causes for policy purposes and how the application of the framework has the potential to increase the validity of LUC models and resolve ILUC and biofuel controversies.« less

The Effects of Stress on Judgement and Decision Making: An Annotated Bibliography.

DTIC Science & Technology

1991-06-01

consideration of these and other results led to the conclusion that peak traffic is the most generalisable environental stressor for ATC’s behaviour...Hemisphere. JANIS, I. L. (1986). Problems of international crisis management in the nuclear age. Journal of Social Issues , 42(2), 201-220. it...applied to the crucial issue as to whether behavior is consistent across different stress situations or whether it is situation specific. . . .Reviewing

Customized Learning: Potential Air Force Applications

DTIC Science & Technology

2011-01-01

Technology in Education, Vol. 41, No. 2, Winter 2008–2009, pp. 143–159. Dee, Lesley, Cristina Devecchi, Lani Florian, and Steve Cochrane, Being, Having and...225–231. Krasner, Steve , ed., Learning Styles, Connecticut State Department of Education, SERC Library, 2002. As of September 30, 2009: http...Training Command, February 18, 2009. Torrance, Harry, Helen Colley, Dean Garratt, Janis Jarvis , Heather Piper, Kathryn Ecclestone, and David James, The

Forecasting the response of Earth's surface to future climatic and land use changes: A review of methods and research needs

DOE PAGES

Pelletier, Jon D.; Murray, A. Brad; Pierce, Jennifer L.; ...

2015-07-14

In the future, Earth will be warmer, precipitation events will be more extreme, global mean sea level will rise, and many arid and semiarid regions will be drier. Human modifications of landscapes will also occur at an accelerated rate as developed areas increase in size and population density. We now have gridded global forecasts, being continually improved, of the climatic and land use changes (C&LUC) that are likely to occur in the coming decades. However, besides a few exceptions, consensus forecasts do not exist for how these C&LUC will likely impact Earth-surface processes and hazards. In some cases, we havemore » the tools to forecast the geomorphic responses to likely future C&LUC. Fully exploiting these models and utilizing these tools will require close collaboration among Earth-surface scientists and Earth-system modelers. This paper assesses the state-of-the-art tools and data that are being used or could be used to forecast changes in the state of Earth's surface as a result of likely future C&LUC. We also propose strategies for filling key knowledge gaps, emphasizing where additional basic research and/or collaboration across disciplines are necessary. The main body of the paper addresses cross-cutting issues, including the importance of nonlinear/threshold-dominated interactions among topography, vegetation, and sediment transport, as well as the importance of alternate stable states and extreme, rare events for understanding and forecasting Earth-surface response to C&LUC. Five supplements delve into different scales or process zones (global-scale assessments and fluvial, aeolian, glacial/periglacial, and coastal process zones) in detail.« less

Development of a recombinant human ovarian (BG1) cell line containing estrogen receptor α and β for improved detection of estrogenic/antiestrogenic chemicals.

PubMed

Brennan, Jennifer C; Bassal, Arzoo; He, Guochun; Denison, Michael S

2016-01-01

Estrogenic endocrine-disrupting chemicals are found in environmental and biological samples, commercial and consumer products, food, and numerous other sources. Given their ubiquitous nature and potential for adverse effects, a critical need exists for rapidly detecting these chemicals. The authors developed an estrogen-responsive recombinant human ovarian (BG1Luc4E2) cell line recently accepted by the US Environmental Protection Agency (USEPA) and Organisation for Economic Co-operation and Development (OECD) as a bioanalytical method to detect estrogen receptor (ER) agonists/antagonists. Unfortunately, these cells appear to contain only 1 of the 2 known ER isoforms, ERα but not ERβ, and the differential ligand selectivity of these ERs indicates that the currently accepted screening method only detects a subset of total estrogenic chemicals. To improve the estrogen screening bioassay, BG1Luc4E2 cells were stably transfected with an ERβ expression plasmid and positive clones identified using ERβ-selective ligands (genistein and Br-ERβ-041). A highly responsive clone (BG1LucERβc9) was identified that exhibited greater sensitivity and responsiveness to ERβ-selective ligands than BG1Luc4E2 cells, and quantitative reverse-transcription polymerase chain reaction confirmed the presence of ERβ expression in these cells. Screening of pesticides and industrial chemicals identified chemicals that preferentially stimulated ERβ-dependent reporter gene expression. Together, these results not only demonstrate the utility of this dual-ER recombinant cell line for detecting a broader range of estrogenic chemicals than the current BG1Luc4E2 cell line, but screening with both cell lines allows identification of ERα- and ERβ-selective chemicals. © 2015 SETAC.

Estimating 20-year land-use change and derived CO2 emissions associated with crops, pasture and forestry in Brazil and each of its 27 states.

PubMed

Novaes, Renan M L; Pazianotto, Ricardo A A; Brandão, Miguel; Alves, Bruno J R; May, André; Folegatti-Matsuura, Marília I S

2017-09-01

Land-use change (LUC) in Brazil has important implications on global climate change, ecosystem services and biodiversity, and agricultural expansion plays a critical role in this process. Concerns over these issues have led to the need for estimating the magnitude and impacts associated with that, which are increasingly reported in the environmental assessment of products. Currently, there is an extensive debate on which methods are more appropriate for estimating LUC and related emissions and regionalized estimates are lacking for Brazil, which is a world leader in agricultural production (e.g. food, fibres and bioenergy). We developed a method for estimating scenarios of past 20-year LUC and derived CO 2 emission rates associated with 64 crops, pasture and forestry in Brazil as whole and in each of its 27 states, based on time-series statistics and in accordance with most used carbon-footprinting standards. The scenarios adopted provide a range between minimum and maximum rates of CO 2 emissions from LUC according to different possibilities of land-use transitions, which can have large impacts in the results. Specificities of Brazil, like multiple cropping and highly heterogeneous carbon stocks, are also addressed. The highest CO 2 emission rates are observed in the Amazon biome states and crops with the highest rates are those that have undergone expansion in this region. Some states and crops showing large agricultural areas have low emissions associated, especially in southern and eastern Brazil. Native carbon stocks and time of agricultural expansion are the most decisive factors to the patterns of emissions. Some implications on LUC estimation methods and standards and on agri-environmental policies are discussed. © 2017 John Wiley & Sons Ltd.

Characterisation of circadian rhythms of various duckweeds.

PubMed

Muranaka, T; Okada, M; Yomo, J; Kubota, S; Oyama, T

2015-01-01

The plant circadian clock controls various physiological phenomena that are important for adaptation to natural day-night cycles. Many components of the circadian clock have been identified in Arabidopsis thaliana, the model plant for molecular genetic studies. Recent studies revealed evolutionary conservation of clock components in green plants. Homologues of clock-related genes have been isolated from Lemna gibba and Lemna aequinoctialis, and it has been demonstrated that these homologues function in the clock system in a manner similar to their functioning in Arabidopsis. While clock components are widely conserved, circadian phenomena display diversity even within the Lemna genus. In order to survey the full extent of diversity in circadian rhythms among duckweed plants, we characterised the circadian rhythms of duckweed by employing a semi-transient bioluminescent reporter system. Using a particle bombardment method, circadian bioluminescent reporters were introduced into nine strains representing five duckweed species: Spirodela polyrhiza, Landoltia punctata, Lemna gibba, L. aequinoctialis and Wolffia columbiana. We then monitored luciferase (luc+) reporter activities driven by AtCCA1, ZmUBQ1 or CaMV35S promoters under entrainment and free-running conditions. Under entrainment, AtCCA1::luc+ showed similar diurnal rhythms in all strains. This suggests that the mechanism of biological timing under day-night cycles is conserved throughout the evolution of duckweeds. Under free-running conditions, we observed circadian rhythms of AtCCA1::luc+, ZmUBQ1::luc+ and CaMV35S::luc+. These circadian rhythms showed diversity in period length and sustainability, suggesting that circadian clock mechanisms are somewhat diversified among duckweeds. © 2014 German Botanical Society and The Royal Botanical Society of the Netherlands.

Functional analysis of the sea urchin-derived arylsulfatase (Ars)-element in mammalian cells.

PubMed

Watanabe, Satoshi; Watanabe, Sachiko; Sakamoto, Naoaki; Sato, Masahiro; Akasaka, Koji

2006-09-01

An insulator is a DNA sequence that has both enhancer-blocking activity, through its ability to modify the influence of neighboring cis-acting elements, and a barrier function that protects a transgene from being silenced by surrounding chromatin. Previously, we isolated and characterized a 582-bp-long element from the sea urchin arylsulfatase gene (Ars). This Ars-element was effective in sea urchin and Drosophila embryos and in plant cells. To investigate Ars-element activity in mammalian cells, we placed the element between the cytomegalovirus enhancer and a luciferase (luc) expression cassette. In contrast to controls lacking the Ars-element, NIH3T3 and 293T cells transfected with the element-containing construct displayed reduced luciferase activities. The Ars-element therefore acts as an enhancer-blocking element in mammalian cells. We assessed the barrier activity of the Ars-element using vectors in which a luc expression cassette was placed between two elements. Transfection experiments demonstrated that luc activity in these vectors was approximately ten-fold higher than in vectors lacking elements. Luc activities were well maintained even after 12 weeks in culture. Our observations demonstrate that the Ars-element has also a barrier activity. These results indicated that the Ars-element act as an insulator in mammalian cells.

Analysis of molecular chaperones using a Xenopus oocyte protein refolding assay.

PubMed

Heikkila, John J; Kaldis, Angelo; Abdulle, Rashid

2006-01-01

Heat shock proteins (Hsps) are molecular chaperones that aid in the folding and translocation of protein under normal conditions and protect cellular proteins during stressful situations. A family of Hsps, the small Hsps, can maintain denatured target proteins in a folding-competent state such that they can be refolded and regain biological activity in the presence of other molecular chaperones. Previous assays have employed cellular lysates as a source of molecular chaperones involved in folding. In this chapter, we describe the production and purification of a Xenopus laevis recombinant small Hsp, Hsp30C, and an in vivo luciferase (LUC) refolding assay employing microinjected Xenopus oocytes. This assay tests whether LUC can be maintained in a folding-competent state when heat denatured in the presence of a small Hsp or other molecular chaperone. For example, micro-injection of heat-denatured LUC alone into oocytes resulted in minimal reactivation of enzyme activity. However, LUC heat denatured in the presence of Hsp30C resulted in 100% recovery of enzyme activity after microinjection. The in vivo oocyte refolding system is more sensitive and requires less molecular chaperone than in vitro refolding assays. Also, this protocol is not limited to testing Xenopus molecular chaperones because small Hsps from other organisms have been used successfully.

A novel dual luciferase assay for the simultaneous monitoring of HIV infection and cell viability.

PubMed

Mitsuki, Yu-Ya; Yamamoto, Takuya; Mizukoshi, Fuminori; Momota, Masatoshi; Terahara, Kazutaka; Yoshimura, Kazuhisa; Harada, Shigeyoshi; Tsunetsugu-Yokota, Yasuko

2016-05-01

Human immunodeficiency virus type 1 (HIV-1) reporter cell lines are critical tools for drug development. However, one disadvantage of HIV-1 reporter cell lines is that reductions in reporter gene activity need to be normalized to cytotoxicity, i.e., live cell numbers. Here, we developed a dual luciferase assay based on a R. reniformis luciferase (hRLuc)-expressing R5-type HIV-1 (NLAD8-hRLuc) and a CEM cell line expressing CCR5 and firefly luciferase (R5CEM-FiLuc). The NLAD8-hRLuc reporter virus was replication competent in peripheral blood mononuclear cells. The level of hRLuc was correlated with p24 antigen levels (p<0.001, R=0.862). The target cell line, R5CEM-FiLuc, stably expressed the firefly luciferase (FiLuc) reporter gene and allowed the simultaneous monitoring of compound cytotoxicity. The dual reporter assay combining a NLAD8-hRLuc virus with R5CEM-FiLuc cells permitted the accurate determination of drug susceptibility for entry, reverse transcriptase, integrase, and protease inhibitors at different multiplicities of infection. This dual reporter assay provides a rapid and direct method for the simultaneous monitoring of HIV infection and cell viability. Copyright © 2016 Elsevier B.V. All rights reserved.

Scientific analysis is essential to assess biofuel policy effects: in response to the paper by Kim and Dale on "Indirect land use change for biofuels: Testing predictions and improving analytical methodologies"

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kline, Keith L; Oladosu, Gbadebo A; Dale, Virginia H

2011-01-01

Vigorous debate on the effects of biofuels derives largely from the changes in land use estimated using economic models designed mainly for the analysis of agricultural trade and markets. The models referenced for land-use change (LUC) analysis in the U.S. Environmental Protection Agency Final Rule on the Renewable Fuel Standard include GTAP, FAPRI-CARD, and FASOM. To address bioenergy impacts, these models were expanded and modified to facilitate simulations of hypothesized LUC. However, even when models use similar basic assumptions and data, the range of LUC results can vary by ten-fold or more. While the market dynamics simulated in these modelsmore » include processes that are important in estimating effects of biofuel policies, the models have not been validated for estimating land-use changes and employ crucial assumptions and simplifications that contradict empirical evidence.« less

Characterization of recombinant Raccoonpox Vaccine Vectors in Chickens

USGS Publications Warehouse

Hwa, S.-H.; Iams, Keith P.; Hall, Jeffrey S.; Kingstad, B.A.; Osorio, Jorge E.

2010-01-01

Raccoonpox virus (RCN) has been used as a recombinant vector against several mammalian pathogens but has not been tested in birds. The replication of RCN in chick embryo fibroblasts (CEFs) and chickens was studied with the use of highly pathogenic avian influenza virus H5N1 hemagglutinin (HA) as a model antigen and luciferase (luc) as a reporter gene. Although RCN replicated to low levels in CEFs, it efficiently expressed recombinant proteins and, in vivo, elicited anti-HA immunoglobulin yolk (IgY) antibody responses comparable to inactivated influenza virus. Biophotonic in vivo imaging of 1-wk-old chicks with RCN-luc showed strong expression of the luc reporter gene lasting up to 3 days postinfection. These studies demonstrate the potential of RCN as a vaccine vector for avian influenza and other poultry pathogens. ?? American Association of Avian Pathologists 2010.

Acts of Atrocity: Effects on Public Opinion Support During War or Conflict

DTIC Science & Technology

2007-12-01

Taylor Martin, “Her Job: Lock Up Iraq’s Bad Guys: Army Brig. General Janis Karpinski, the Lone Female Commander in Iraq, Runs the Prison System that...photographing naked male and female detainees; forcibly arranging detainees in various sexually explicit positions for photographing; forcing...a dog chain or strap around a naked detainee’s neck and having a female soldier pose for a picture; a male MP guard having sex with a female

Drivers of potential GHG fluxes under bioenergy land use change in the UK

NASA Astrophysics Data System (ADS)

Parmar, Kim; Keith, Aidan M.; Perks, Mike; Rowe, Rebecca; Sohi, Saran; McNamara, Niall

2013-04-01

The greatest contributors to global greenhouse gases (GHG's) are CO2 emissions from fossil fuel use and following land use change (LUC). Globally, soils contain three times more carbon than the atmosphere and have the potential to act as GHG sources or sinks. A significant amount of land may be converted to bioenergy production to help meet UK 2050 renewable energy and GHG emissions reduction targets. This raises considerable sustainability concerns with respect to the effects of LUC on soil carbon (C) conservation and GHG emissions. Forests are a key component in the global C cycle and when managed effectively can reduce atmospheric GHG concentrations. Together with other dedicated bioenergy crops, Short Rotation Forestry (SRF) could be used to meet biomass requirements. SRF is defined as high density plantations of fastgrowing tree species grown on short rotational lengths (8-20 years) for biomass (McKay 2011). As SRF is likely to be an important domestic source of biomass for energy it is imperative that we gain an understanding of the implications for large-scale commercial application on soil C and the GHG balance. We utilized a paired-site approach to investigate how LUC to SRF could potentially alter the underlying processes of soil GHG production and consumption. This work was linked to a wider soil C stock inventory for bioenergy LUC, so our major focus was on changes to soil respiration. Specifically, we examined the relative importance of litter, soil, and microbial properties in determining potential soil respiration, and whether these relationships were consistent at different soil temperatures (10 ° C and 20 ° C). Soils were sampled to a depth of 30 cm from 30 LUC transitions across the UK and incubated under controlled laboratory conditions, with gas samples taken over a seven day enclosure period. CO2, N2O and CH4 gas fluxes were measured by gas chromatography and were examined together with other soil properties measured in the field and

Gaining ground in the modeling of land-use change greenhouse gas emissions associated with biofuel production

NASA Astrophysics Data System (ADS)

Dunn, J.; Mueller, S.; Kwon, H.; Wang, M.; Wander, M.

2012-12-01

Land-use change (LUC) resulting from biofuel feedstock production and the associated greenhouse gas (GHG) emissions are a hotly-debated aspect of biofuels. Certainly, LUC GHG emissions are one of the most uncertain elements in life cycle analyses (LCA) of biofuels. To estimate LUC GHG emissions, two sets of data are necessary. First, information on the amount and type of land that is converted to biofuel feedstock production is required. These data are typically generated through application of computable general equilibrium (CGE) models such as Purdue University's Global Trade Analysis Project (GTAP) model. Second, soil carbon content data for the affected land types is essential. Recently, Argonne National Laboratory's Carbon Calculator for Land Use Change from Biofuels Production (CCLUB) has been updated with CGE modeling results that estimate the amount and type of LUC world-wide from production of ethanol from corn, corn stover, miscanthus, and switchgrass (Mueller et al. 2012). Moreover, we have developed state-specific carbon content data, determined through modeling with CENTURY, for the two most dominant soil types in the conterminous 48 U.S. states (Kwon et al. 2012) to enable finer-resolution results for domestic LUC GHG emissions for these ethanol production scenarios. Of the feedstocks examined, CCLUB estimates that LUC GHG emissions are highest for corn ethanol (9.1 g CO2e/MJ ethanol) and lowest for miscanthus (-12 g CO2e/MJ ethanol). We will present key observations from CCLUB results incorporated into Argonne National Laboratory's Greenhouse Gases, Regulated Emissions, and Energy use in Transportation (GREET) model, which is a LCA tool for transportation fuels and advanced vehicle technologies. We will discuss selected issues in this modeling, including the sensitivity of domestic soil carbon emission factors to modeling parameters and assumptions about the fate of harvested wood products. Further, we will discuss efforts to update CCLUB with county

In vivo visualization and ex vivo quantification of murine breast cancer cells in the mouse brain using MRI cell tracking and electron paramagnetic resonance.

PubMed

Danhier, Pierre; Magat, Julie; Levêque, Philippe; De Preter, Géraldine; Porporato, Paolo E; Bouzin, Caroline; Jordan, Bénédicte F; Demeur, Gladys; Haufroid, Vincent; Feron, Olivier; Sonveaux, Pierre; Gallez, Bernard

2015-03-01

Cell tracking could be useful to elucidate fundamental processes of cancer biology such as metastasis. The aim of this study was to visualize, using MRI, and to quantify, using electron paramagnetic resonance (EPR), the entrapment of murine breast cancer cells labeled with superparamagnetic iron oxide particles (SPIOs) in the mouse brain after intracardiac injection. For this purpose, luciferase-expressing murine 4 T1-luc breast cancer cells were labeled with fluorescent Molday ION Rhodamine B SPIOs. Following intracardiac injection, SPIO-labeled 4 T1-luc cells were imaged using multiple gradient-echo sequences. Ex vivo iron oxide quantification in the mouse brain was performed using EPR (9 GHz). The long-term fate of 4 T1-luc cells after injection was characterized using bioluminescence imaging (BLI), brain MRI and immunofluorescence. We observed hypointense spots due to SPIO-labeled cells in the mouse brain 4 h after injection on T2 *-weighted images. Histology studies showed that SPIO-labeled cancer cells were localized within blood vessels shortly after delivery. Ex vivo quantification of SPIOs showed that less than 1% of the injected cells were taken up by the mouse brain after injection. MRI experiments did not reveal the development of macrometastases in the mouse brain several days after injection, but immunofluorescence studies demonstrated that these cells found in the brain established micrometastases. Concerning the metastatic patterns of 4 T1-luc cells, an EPR biodistribution study demonstrated that SPIO-labeled 4 T1-luc cells were also entrapped in the lungs of mice after intracardiac injection. BLI performed 6 days after injection of 4 T1-luc cells showed that this cell line formed macrometastases in the lungs and in the bones. Conclusively, EPR and MRI were found to be complementary for cell tracking applications. MRI cell tracking at 11.7 T allowed sensitive detection of isolated SPIO-labeled cells in the mouse brain, whereas EPR

microRNA-Based Immunotherapy for Control of Early Stage Lung Cancer

DTIC Science & Technology

2016-09-01

in NSG hosts via subcutaneously injection. A549-Luc developed tumors successfully in NSG hosts and mice bearing A549-Luc tumors were the subject...activation of NK cells from whole blood. Next we evaluated NK cells and host interaction by transferring NK cells into A549-tumor bearing NSG host via...that did not receive NK cells. 4 At day 28, we harvested tumors, blood and tissues from tumor- bearing mice to analyze for NK presence in the

Molecular Determinants of Antiestrogen and Drug Sensitivity in Breast Carcinoma Cells

DTIC Science & Technology

1997-08-01

W.S. human HT1080 fibrosarcoma cells (a gift of Pear, personal communication). G.R. Stark), susceptible to ecotropic retrovirus In several...transfection with equal amounts of plasmid pOPRSVIluc (generated by replacing CAT of pORRSVICAT with luc) which expresses luc from R02 promoter, and control...retroviral transduction into (Fig. 1). During transient co-transfection of NIH 3T3 human HT1080 fibrosarcoma or mouse NIH 3T3 cell cells with a laac expressing

In vitro modeling of HIV proviral activity in microglia.

PubMed

Campbell, Lee A; Richie, Christopher T; Zhang, Yajun; Heathward, Emily J; Coke, Lamarque M; Park, Emily Y; Harvey, Brandon K

2017-12-01

Microglia, the resident macrophages of the brain, play a key role in the pathogenesis of HIV-associated neurocognitive disorders (HAND) due to their productive infection by HIV. This results in the release of neurotoxic viral proteins and pro-inflammatory compounds which negatively affect the functionality of surrounding neurons. Because models of HIV infection within the brain are limited, we aimed to create a novel microglia cell line with an integrated HIV provirus capable of recreating several hallmarks of HIV infection. We utilized clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 gene editing technology and integrated a modified HIV provirus into CHME-5 immortalized microglia to create HIV-NanoLuc CHME-5. In the modified provirus, the Gag-Pol region is replaced with the coding region for NanoLuciferase (NanoLuc), which allows for the rapid assay of HIV long terminal repeat activity using a luminescent substrate, while still containing the necessary genetic material to produce established neurotoxic viral proteins (e.g. tat, nef, gp120). We confirmed that HIV-NanoLuc CHME-5 microglia express NanoLuc, along with the HIV viral protein Nef. We subsequently exposed these cells to a battery of experiments to modulate the activity of the provirus. Proviral activity was enhanced by treating the cells with pro-inflammatory factors lipopolysaccharide (LPS) and tumor necrosis factor alpha and by overexpressing the viral regulatory protein Tat. Conversely, genetic modification of the toll-like receptor-4 gene by CRISPR/Cas9 reduced LPS-mediated proviral activation, and pharmacological application of NF-κB inhibitor sulfasalazine similarly diminished proviral activity. Overall, these data suggest that HIV-NanoLuc CHME-5 may be a useful tool in the study of HIV-mediated neuropathology and proviral regulation. Published 2017. This article is a U.S. Government work and is in the public domain in the USA.

A synthetic luciferin improves in vivo bioluminescence imaging of gene expression in cardiovascular brain regions.

PubMed

Simonyan, Hayk; Hurr, Chansol; Young, Colin N

2016-10-01

Bioluminescence imaging is an effective tool for in vivo investigation of molecular processes. We have demonstrated the applicability of bioluminescence imaging to spatiotemporally monitor gene expression in cardioregulatory brain nuclei during the development of cardiovascular disease, via incorporation of firefly luciferase into living animals, combined with exogenous d-luciferin substrate administration. Nevertheless, d-luciferin uptake into the brain tissue is low, which decreases the sensitivity of bioluminescence detection, particularly when considering small changes in gene expression in tiny central areas. Here, we tested the hypothesis that a synthetic luciferin, cyclic alkylaminoluciferin (CycLuc1), would be superior to d-luciferin for in vivo bioluminescence imaging in cardiovascular brain regions. Male C57B1/6 mice underwent targeted delivery of an adenovirus encoding the luciferase gene downstream of the CMV promoter to the subfornical organ (SFO) or paraventricular nucleus of hypothalamus (PVN), two crucial cardioregulatory neural regions. While bioluminescent signals could be obtained following d-luciferin injection (150 mg/kg), CycLuc1 administration resulted in a three- to fourfold greater bioluminescent emission from the SFO and PVN, at 10- to 20-fold lower substrate concentrations (7.5-15 mg/kg). This CycLuc1-mediated enhancement in bioluminescent emission was evident early following substrate administration (i.e., 6-10 min) and persisted for up to 1 h. When the exposure time was reduced from 60 s to 1,500 ms, minimal signal in the PVN was detectable with d-luciferin, whereas bioluminescent images could be reliably captured with CycLuc1. These findings demonstrate that bioluminescent imaging with the synthetic luciferin CycLuc1 provides an improved physiological genomics tool to investigate molecular events in discrete cardioregulatory brain nuclei. Copyright © 2016 the American Physiological Society.

Lentivirus-mediated bifunctional cell labeling for in vivo melanoma study

PubMed Central

Day, Chi-Ping; Carter, John; Bonomi, Carrie; Esposito, Dominic; Crise, Bruce; Ortiz-Conde, Betty; Hollingshead, Melinda; Merlino, Glenn

2009-01-01

SUMMARY Lentiviral vectors (LVs) are capable of labeling a broad spectrum of cell types, achieving stable expression of transgenes. However, for in vivo studies, the duration of marker gene expression has been highly variable. We have developed a series of LVs harboring different promoters for expressing reporter gene in mouse cells. Long-term culture and colony formation of several LV-labeled mouse melanoma cells showed that promoters derived from mammalian house-keeping genes, especially those encoding RNA polymerase II (Pol2) and ferritin (FerH), provided the highest consistency for reporter expression. For in vivo studies, primary B16BL6 mouse melanoma were infected with LVs whose luciferase-GFP fusion gene (Luc/GFP) was driven by either Pol2 or FerH promoters. When transplanted into syngeneic C57BL/6 mice, Luc/GFP-labeled B16BL6 mouse melanoma cells can be monitored by bioluminescence imaging in vivo, and GFP-positive cells can be isolated from the tumors by FACS. Pol2-Luc/GFP labeling, while lower in activity, was more sustainable than FerH-Luc/GFP labeling in B16BL6 over consecutive passages into mice. We conclude that Pol-2-Luc/GFP labeling allows long-term in vivo monitoring and tumor cell isolation in immunocompetent mouse melanoma models. SIGNIFICANCE In this study we have developed and identified lentiviral vectors that allow labeled mouse melanoma cells to maintain long-term and consistent expression of a bifunctional luciferase-GFP marker gene, even in syngeneic mice with an intact immune function. This cell-labeling system can be used to build immunocompetent mouse melanoma models that permit both tumor monitoring and FACS-based tumor cell isolation from tissues, greatly facilitating the in vivo study of melanoma. PMID:19175523

Human induced pluripotent stem cell-derived cardiomyocytes as an in vitro model for coxsackievirus B3-induced myocarditis and antiviral drug screening platform.

PubMed

Sharma, Arun; Marceau, Caleb; Hamaguchi, Ryoko; Burridge, Paul W; Rajarajan, Kuppusamy; Churko, Jared M; Wu, Haodi; Sallam, Karim I; Matsa, Elena; Sturzu, Anthony C; Che, Yonglu; Ebert, Antje; Diecke, Sebastian; Liang, Ping; Red-Horse, Kristy; Carette, Jan E; Wu, Sean M; Wu, Joseph C

2014-08-29

Viral myocarditis is a life-threatening illness that may lead to heart failure or cardiac arrhythmias. A major causative agent for viral myocarditis is the B3 strain of coxsackievirus, a positive-sense RNA enterovirus. However, human cardiac tissues are difficult to procure in sufficient enough quantities for studying the mechanisms of cardiac-specific viral infection. This study examined whether human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) could be used to model the pathogenic processes of coxsackievirus-induced viral myocarditis and to screen antiviral therapeutics for efficacy. hiPSC-CMs were infected with a luciferase-expressing coxsackievirus B3 strain (CVB3-Luc). Brightfield microscopy, immunofluorescence, and calcium imaging were used to characterize virally infected hiPSC-CMs for alterations in cellular morphology and calcium handling. Viral proliferation in hiPSC-CMs was quantified using bioluminescence imaging. Antiviral compounds including interferonβ1, ribavirin, pyrrolidine dithiocarbamate, and fluoxetine were tested for their capacity to abrogate CVB3-Luc proliferation in hiPSC-CMs in vitro. The ability of these compounds to reduce CVB3-Luc proliferation in hiPSC-CMs was consistent with reported drug effects in previous studies. Mechanistic analyses via gene expression profiling of hiPSC-CMs infected with CVB3-Luc revealed an activation of viral RNA and protein clearance pathways after interferonβ1 treatment. This study demonstrates that hiPSC-CMs express the coxsackievirus and adenovirus receptor, are susceptible to coxsackievirus infection, and can be used to predict antiviral drug efficacy. Our results suggest that the hiPSC-CM/CVB3-Luc assay is a sensitive platform that can screen novel antiviral therapeutics for their effectiveness in a high-throughput fashion. © 2014 American Heart Association, Inc.

Mapping hydrological signatures in the tropical Andes using a network of paired catchments

NASA Astrophysics Data System (ADS)

Ochoa-Tocachi, B. F.; Buytaert, W.; De Bièvre, B.

2016-12-01

The complexity and data scarcity of tropical Andean catchments make regional hydrological predictions very challenging. The strong spatiotemporal patterns of the local climate contrast with the inadequate coverage, especially of remote areas, by the national monitoring networks. We present an approach to regionalize the hydrological impacts of land-use and land-cover (LUC) using a network of 24 headwater catchments in a pairwise comparison approach. We monitored precipitation and streamflow through an informal partnership of stakeholders in the Andes, known as iMHEA. Using a `trading-space-for-time' approach, our design aims at strengthening the statistical significance of LUC signals. To test our hypothesis, we summarized the hydrological responses using a set of indices, which are then regionalized against catchment properties including land-use. Lastly, the regionalization model is then used to generate distributed maps of hydrological signatures in ungauged areas. Our results clearly reflect the dominant regional climate patterns of the tropical Andes and the associated wide spectrum of hydrological responses. Although the hydrological impacts of LUC are equally diverse, we find consistent trends within different biomes. Contrary to earlier studies, we find that incorporating LUC variables in the regionalization increases significantly the performance of the regression model and its predictive capacity, which makes it possible to generate regional maps that predict the dynamics and propagation of streamflow signatures in complex regions with an explicit report of uncertainty. We attribute the robust regionalization results to the regional pairwise setup that covers diverse physiographic characteristics, contrasting LUC types, and degrees of conservation/alteration. As such, it may be a useful strategy to optimize data collection, leverage commonly available geographical information, and understand the major controls of hydrological response in data

An audit of the global carbon budget: identifying and reducing sources of uncertainty

NASA Astrophysics Data System (ADS)

Ballantyne, A. P.; Tans, P. P.; Marland, G.; Stocker, B. D.

2012-12-01

Uncertainties in our carbon accounting practices may limit our ability to objectively verify emission reductions on regional scales. Furthermore uncertainties in the global C budget must be reduced to benchmark Earth System Models that incorporate carbon-climate interactions. Here we present an audit of the global C budget where we try to identify sources of uncertainty for major terms in the global C budget. The atmospheric growth rate of CO2 has increased significantly over the last 50 years, while the uncertainty in calculating the global atmospheric growth rate has been reduced from 0.4 ppm/yr to 0.2 ppm/yr (95% confidence). Although we have greatly reduced global CO2 growth rate uncertainties, there remain regions, such as the Southern Hemisphere, Tropics and Arctic, where changes in regional sources/sinks will remain difficult to detect without additional observations. Increases in fossil fuel (FF) emissions are the primary factor driving the increase in global CO2 growth rate; however, our confidence in FF emission estimates has actually gone down. Based on a comparison of multiple estimates, FF emissions have increased from 2.45 ± 0.12 PgC/yr in 1959 to 9.40 ± 0.66 PgC/yr in 2010. Major sources of increasing FF emission uncertainty are increased emissions from emerging economies, such as China and India, as well as subtle differences in accounting practices. Lastly, we evaluate emission estimates from Land Use Change (LUC). Although relative errors in emission estimates from LUC are quite high (2 sigma ~ 50%), LUC emissions have remained fairly constant in recent decades. We evaluate the three commonly used approaches to estimating LUC emissions- Bookkeeping, Satellite Imagery, and Model Simulations- to identify their main sources of error and their ability to detect net emissions from LUC.; Uncertainties in Fossil Fuel Emissions over the last 50 years.

The Major Portal of Entry of Koi Herpesvirus in Cyprinus carpio Is the Skin▿

PubMed Central

Costes, B.; Raj, V. Stalin; Michel, B.; Fournier, G.; Thirion, M.; Gillet, L.; Mast, J.; Lieffrig, F.; Bremont, M.; Vanderplasschen, A.

2009-01-01

Koi herpesvirus (KHV), recently designated Cyprinid herpesvirus 3, is the causative agent of a lethal disease in koi and common carp. In the present study, we investigated the portal of entry of KHV in carp by using bioluminescence imaging. Taking advantage of the recent cloning of the KHV genome as a bacterial artificial chromosome (BAC), we produced a recombinant plasmid encoding a firefly luciferase (LUC) expression cassette inserted in the intergenic region between open reading frame (ORF) 136 and ORF 137. Two viral strains were then reconstituted from the modified plasmid, the FL BAC 136 LUC excised strain and the FL BAC 136 LUC TK revertant strain, including a disrupted and a wild-type thymidine kinase (TK) locus, respectively. In vitro, the two recombinant strains replicated comparably to the parental FL strain. The FL BAC 136 LUC TK revertant strain was shown in vitro to induce a bioluminescent signal allowing the detection of single positive cells as early as 24 h postinfection, while in vivo, it induced KHV infection in carp that was indistinguishable from that induced by the parental FL strain. To identify the KHV portal of entry, carp were analyzed by bioluminescence imaging at different times postinfection with the FL BAC 136 LUC TK revertant strain. These analyses demonstrated that the skin of the fish covering the fins and also the body is the major portal of entry for KHV in carp. Finally, to further demonstrate the role of the skin as the KHV portal of entry, we constructed an original system, nicknamed “U-tube,” to perform percutaneous infection restricted to the posterior part of the fish. All the data obtained in the present study demonstrate that the skin, and not the gills, is the major portal of entry for KHV in carp. PMID:19153228

How can land-use modelling tools inform bioenergy policies?

PubMed Central

Davis, Sarah C.; House, Joanna I.; Diaz-Chavez, Rocio A.; Molnar, Andras; Valin, Hugo; DeLucia, Evan H.

2011-01-01

Targets for bioenergy have been set worldwide to mitigate climate change. Although feedstock sources are often ambiguous, pledges in European nations, the United States and Brazil amount to more than 100 Mtoe of biorenewable fuel production by 2020. As a consequence, the biofuel sector is developing rapidly, and it is increasingly important to distinguish bioenergy options that can address energy security and greenhouse gas mitigation from those that cannot. This paper evaluates how bioenergy production affects land-use change (LUC), and to what extent land-use modelling can inform sound decision-making. We identified local and global internalities and externalities of biofuel development scenarios, reviewed relevant data sources and modelling approaches, identified sources of controversy about indirect LUC (iLUC) and then suggested a framework for comprehensive assessments of bioenergy. Ultimately, plant biomass must be managed to produce energy in a way that is consistent with the management of food, feed, fibre, timber and environmental services. Bioenergy production provides opportunities for improved energy security, climate mitigation and rural development, but the environmental and social consequences depend on feedstock choices and geographical location. The most desirable solutions for bioenergy production will include policies that incentivize regionally integrated management of diverse resources with low inputs, high yields, co-products, multiple benefits and minimal risks of iLUC. Many integrated assessment models include energy resources, trade, technological development and regional environmental conditions, but do not account for biodiversity and lack detailed data on the location of degraded and underproductive lands that would be ideal for bioenergy production. Specific practices that would maximize the benefits of bioenergy production regionally need to be identified before a global analysis of bioenergy-related LUC can be accomplished. PMID

A key general stress response motif is regulated non-uniformly by CAMTA transcription factors.

PubMed

Benn, Geoffrey; Wang, Chang-Quan; Hicks, Derrick R; Stein, Jeffrey; Guthrie, Cade; Dehesh, Katayoon

2014-10-01

Plants cope with environmental challenges by rapidly triggering and synchronizing mechanisms governing stress-specific and general stress response (GSR) networks. The GSR acts rapidly and transiently in response to various stresses, but the underpinning mechanisms have remained elusive. To define GSR regulatory components we have exploited the Rapid Stress Response Element (RSRE), a previously established functional GSR motif, using Arabidopsis plants expressing a 4xRSRE::Luciferase (RSRE::LUC) reporter. Initially, we searched public microarray datasets and found an enrichment of RSRE in promoter sequences of stress genes. Next, we treated RSRE::LUC plants with wounding and a range of rapidly stress-inducible hormones and detected a robust LUC activity solely in response to wounding. Application of two Ca(2+) burst inducers, flagellin22 (flg22) and oligogalacturonic acid, activated RSRE strongly and systemically, while the Ca(2+) chelator ethylene glycol tetraacetic acid (EGTA) significantly reduced wound induction of RSRE::LUC. In line with the signaling function of Ca(2+) in transduction events leading to activation of RSRE, we examined the role of CALMODULIN-BINDING TRANSCRIPTIONAL ACTIVATORs (CAMTAs) in RSRE induction. Transient expression assays displayed CAMTA3 induction of RSRE and not that of the mutated element mRSRE. Treatment of selected camta mutant lines integrated into RSRE::LUC parent plant, with wounding, flg22, and freezing, established a differential function of these CAMTAs in potentiating the activity of RSRE. Wound response studies using camta double mutants revealed a cooperative function of CAMTAs2 and 4 with CAMTA 3 in the RSRE regulation. These studies provide insights into governing components of transduction events and reveal transcriptional modules that tune the expression of a key GSR motif. © 2014 The Authors The Plant Journal © 2014 John Wiley & Sons Ltd.

Computational investigation of the effect of pH on the color of firefly bioluminescence by DFT.

PubMed

Pinto da Silva, Luís; Esteves da Silva, Joaquim C G

2011-04-04

In spite of recent advances towards understanding the mechanism of firefly bioluminescence, there is no consensus about which oxyluciferin (OxyLH(2)) species are the red and yellow-green emitters. The crystal structure of Luciola cruciata luciferase (LcLuc) revealed different conformations for the various steps of the bioluminescence reaction, with different degrees of polarity and rigidity of the active-site microenvironment. In this study, these different conformations of luciferase (Luc) are simulated and their effects on the different chemical equilibria of OxyLH(2) are investigated as a function of pH by means of density functional theory with the PBE0 functional. In particular, the thermodynamic properties and the absorption spectra of each species, as well as their relative stabilities in the ground and excited states, were computed in the different conformations of Luc. From the calculations it is possible to derive the acid dissociation and tautomeric constants, and the corresponding distribution diagrams. It is observed that the anionic keto form of OxyLH(2) is both the red and the yellow-green emitter. Consequently, the effect of Luc conformations on the structural and electronic properties of the Keto-(-1) form are studied. Finally, insights into the Luc-catalyzed light-emitting reaction are derived from the calculations. The multicolor bioluminescence can be explained by interactions of the emitter with active-site molecules, the effects of which on light emission are modulated by the internal dielectric constant of the different conformations. These interactions can suffer also from rearrangement due to entry of external solvent and changes in the protonation state of some amino acid residues and adenosine monophosphate (AMP). Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Attributing land-use change carbon emissions to exported biomass

DOE Office of Scientific and Technical Information (OSTI.GOV)

Saikku, Laura, E-mail: laura.saikku@helsinki.fi; Soimakallio, Sampo, E-mail: sampo.soimakallio@vtt.fi; Pingoud, Kim, E-mail: kim.pingoud@vtt.fi

2012-11-15

In this study, a simple, transparent and robust method is developed in which land-use change (LUC) emissions are retrospectively attributed to exported biomass products based on the agricultural area occupied for the production. LUC emissions account for approximately one-fifth of current greenhouse gas emissions. Increasing agricultural exports are becoming an important driver of deforestation. Brazil and Indonesia are used as case studies due to their significant deforestation in recent years. According to our study, in 2007, approximately 32% and 15% of the total agricultural land harvested and LUC emissions in Brazil and Indonesia respectively were due to exports. The mostmore » important exported single items with regard to deforestation were palm oil for Indonesia and bovine meat for Brazil. To reduce greenhouse gas (GHG) emissions effectively worldwide, leakage of emissions should be avoided. This can be done, for example, by attributing embodied LUC emissions to exported biomass products. With the approach developed in this study, controversial attribution between direct and indirect LUC and amortization of emissions over the product life cycle can be overcome, as the method operates on an average basis and annual level. The approach could be considered in the context of the UNFCCC climate policy instead of, or alongside with, other instruments aimed at reducing deforestation. However, the quality of the data should be improved and some methodological issues, such as the allocation procedure in multiproduct systems and the possible dilution effect through third parties not committed to emission reduction targets, should be considered. - Highlights: Black-Right-Pointing-Pointer CO{sub 2} emissions from land use changes are highly important. Black-Right-Pointing-Pointer Attribution of land use changes for products is difficult. Black-Right-Pointing-Pointer Simple and robust method is developed to attribute land use change emissions.« less

Evaluating reporter genes of different luciferases for optimized in vivo bioluminescence imaging of transplanted neural stem cells in the brain.

PubMed

Mezzanotte, Laura; Aswendt, Markus; Tennstaedt, Annette; Hoeben, Rob; Hoehn, Mathias; Löwik, Clemens

2013-01-01

Bioluminescence imaging (BLI) has become the method of choice for optical tracking of cells in small laboratory animals. However, the use of luciferases from different species, depending on different substrates and emitting at distinct wavelengths, has not been optimized for sensitive neuroimaging. In order to identify the most suitable luciferase, this quantitative study compared the luciferases Luc2, CBG99, PpyRE9 and hRluc. Human embryonic kidney (HEK-293) cells and mouse neural stem cells were transduced by lentiviral vector-mediated transfer to express one of the four luciferases, together with copGFP. A T2A peptide linker promoted stoichiometric expression between both imaging reporters and the comparison of cell populations upon flow cytometry. Cell dilution series were used to determine highest BLI sensitivity in vitro for Luc2. However, Coelenterazine h-dependent hRluc signals clearly exceeded d-luciferin-dependent BLI in vitro. For the quantitative in vivo analysis, cells were transplanted into mouse brain and BLI was performed including the recording of emission kinetics and spectral characteristics. Differences in light kinetics were observed for d-luciferin vs Coelenterazine h. The emission spectra of Luc2 and PpyRE9 remained almost unchanged, while the emission spectrum of CBG99 became biphasic. Most importantly, photon emission decreased in the order of Luc2, CBG99, PpyRE9 to hRluc. The feasibility of combining different luciferases for dual color and dual substrate neuroimaging was tested and discussed. This investigation provides the first complete quantitative comparison of different luciferases expressed by neural stem cells. It results in a clear recommendation of Luc2 as the best luciferase selection for in vivo neuroimaging. Copyright © 2013 John Wiley & Sons, Ltd.

Development and evaluation of yeast-based GFP and luciferase reporter assays for chemical-induced genotoxicity and oxidative damage.

PubMed

Suzuki, Hajime; Sakabe, Takahiro; Hirose, Yuu; Eki, Toshihiko

2017-01-01

We aimed to develop the bioassays for genotixicity and/or oxidative damage using the recombinant yeast. A genotoxicity assay was developed using recombinant Saccharomyces cerevisiae strain BY4741 with a green fluorescent protein (GFP) reporter plasmid, driven by the DNA damage-responsive RNR3 promoter. Enhanced fluorescence induction was observed in DNA repair-deficient strains treated with methyl methanesulfonate, but not with hydrogen peroxide. A GFP reporter yeast strain driven by the oxidative stress-responsive TRX2 promoter was newly developed to assess oxidative damage, but fluorescence was poorly induced by oxidants. In place of GFP, yeast strains with luciferase gene reporter plasmids (luc2 and luc2CP, encoding stable and unstable luciferase, respectively) were prepared. Transient induction of luciferase activity was clearly detected only in a TRX2 promoter-driven luc2CP reporter strain within 90 min of oxidant exposure. However, luciferase was strongly induced by hydroxyurea in the RNR3 promoter-driven luc2 and GFP reporter strains over 8 h after the exposure, suggesting that the RNR3 promoter is continuously upregulated by DNA damage, whereas the TRX2 promoter is transiently activated by oxidative agents. Luciferase activity levels were also increased in a TRX2-promoter-driven luc2CP reporter strain treated with tert-butyl hydroperoxide and menadione and weakly induced with diamide and diethyl maleate. Weakly enhanced luciferase activity induction was detected in the sod1Δ, sod2Δ, and rad27Δ strains treated with hydrogen peroxide compared with that in the wild-type strain. In conclusion, tests using GFP and stable luciferase reporters are useful for genotoxicity, and oxidative damage can be clearly detected by assay with an unstable luciferase reporter.

Climate extremes, land–climate feedbacks and land-use forcing at 1.5°C

DOE Office of Scientific and Technical Information (OSTI.GOV)

Seneviratne, Sonia I.; Wartenburger, Richard; Guillod, Benoit P.

This article investigates projected changes in temperature and water cycle extremes at 1.5°C global warming, and highlights the role of land processes and land-use changes (LUC) for these projections. We provide new comparisons of changes in climate at 1.5°C vs 2°C based on empirical sampling analyses of transient simulations vs simulations from the 'Half a degree Additional warming, Prognosis and Projected Impacts' (HAPPI) multi-model experiment. The two approaches yield overall similar results regarding changes in climate extremes on land, and reveal a substantial difference in regional extremes occurrence at 1.5°C vs 2°C. Land processes mediated through soil moisture feedbacks andmore » land-use forcing play a major role for projected changes in extremes at 1.5°C in most mid-latitude regions, including densely populated areas in North America, Europe and Asia. This has important implications for low-emissions scenarios derived from Integrated Assessment Models (IAMs), which include major LUC in ambitious mitigation pathways (e.g. associated with increased bioenergy use), but are also shown to differ in the simulated LUC patterns. Biogeophysical effects from LUC are not considered in the development of IAM scenarios, but play an important role for projected regional changes in climate extremes, and are thus of high relevance for sustainable development pathways.« less

Hypoxia/hepatoma dual specific suicide gene expression plasmid delivery using bio-reducible polymer for hepatocellular carcinoma therapy.

PubMed

Kim, Hyun Ah; Nam, Kihoon; Lee, Minhyung; Kim, Sung Wan

2013-10-10

Gene therapy is suggested as a promising alternative strategy of hepatocellular carcinoma (HCC, also called hepatoma) therapy. To achieve a successful and safe gene therapy, tight regulation of gene expression is required to minimize side-effects in normal tissues. In this study, we developed a novel hypoxia and hepatoma dual specific gene expression vector. The constructed vectors were transfected into various cell lines using bio-reducible polymer, PAM-ABP. First, pAFPS-Luc or pAFPL-Luc vector was constructed with the alpha-fectoprotein (AFP) promoter and enhancer for hepatoma tissue specific gene expression. Then, pEpo-AFPL-Luc was constructed by insertion of the erythropoietin (Epo) enhancer for hypoxic cancer specific gene expression. In vitro transfection assay showed that pEpo-AFPL-Luc transfected hepatoma cell increased gene expression under hypoxic condition. To confirm the therapeutic effect of dual specific vector, herpes simplex virus thymidine kinase (HSV-TK) gene was introduced for cancer cell killing. The pEpo-AFPL-TK was transfected into hepatoma cell lines in the presence of ganciclovir (GCV) pro-drug. Caspase-3/7, MTT and TUNEL assays elucidated that pEpo-AFPL-TK transfected cells showed significant increasing of death rate in hypoxic hepatoma cells compared to controls. Therefore, the hypoxia/hepatoma dual specific gene expression vector with the Epo enhancer and AFP promoter may be useful for hepatoma specific gene therapy. © 2013.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Seneviratne, Sonia I.; Wartenburger, Richard; Guillod, Benoit

This article investigates projected changes in temperature and water cycle extremes at 1.5°C global warming, and highlights the role of land processes and land-use changes (LUC) for these projections. We provide new comparisons of changes in climate at 1.5°C vs 2°C based on empirical sampling analyses of transient simulations vs simulations from the “Half a degree Additional warming, Prognosis and Projected Impacts” (HAPPI) multi-model experiment. The two approaches yield overall similar results regarding changes in climate extremes on land, and reveal a substantial difference in regional extremes occurrence at 1.5°C vs 2°C. Land processes mediated through soil moisture feedbacks andmore » land-use forcing play a major role for projected changes in extremes at 1.5°C in most mid-latitude regions, including densely populated areas in North America, Europe and Asia. This has important implications for low-emissions scenarios derived from Integrated Assessment Models (IAMs), which include major LUC in ambitious mitigation pathways (e.g. associated with increased bioenergy use), but are also shown to differ in the simulated LUC patterns. Biogeophysical effects from LUC are not considered in the development of IAM scenarios, but play an important role for projected regional changes in climate extremes, and are thus of high relevance for sustainable development pathways.« less

Enzyme-synthesized Poly(amine-co-esters) as Non-viral Vectors for Gene Delivery

PubMed Central

Liu, Jie; Jiang, Zhaozhong; Zhou, Jiangbing; Zhang, Shengmin; Saltzman, W. Mark

2010-01-01

A family of biodegradable poly(amine-co-esters) was synthesized in one step via enzymatic copolymerization of diesters with amino-substituted diols. Diesters of length C4–C12 (i.e., from succinate to dodecanedioate) were successfully copolymerized with diethanolamines with either an alkyl (methyl, ethyl, n-butyl, t-butyl) or an aryl (phenyl) substituent on the nitrogen. Upon protonation at slightly acidic conditions, these poly(amine-co-esters) readily turned to cationic polyelectrolytes, which were capable of condensing with polyanionic DNA to form nanometer-sized polyplexes. In vitro screening with pLucDNA revealed that two of the copolymers, poly(N-methyldiethyleneamine sebacate) (PMSC) and poly(N-ethyldiethyleneamine sebacate) (PESC), possessed comparable or higher transfection efficiencies compared to Lipofectamine 2000. PMSC/pLucDNA and PESC/pLucDNA nanoparticles had desirable particle sizes (40–70 nm) for cellular uptake and were capable of functioning as proton sponges to facilitate endosomal escape after cellular uptake. These polyplex nanoparticles exhibited extremely low cytotoxicity. Furthermore, in vivo gene transfection experiments revealed that PMSC is a substantially more effective gene carrier than PEI in delivering pLucDNAto cells in tumors in mice. All these properties suggest that poly(amine-co-esters) are promising non-viral vectors for safe and efficient DNA delivery in gene therapy. PMID:21171165

Development of an Ultra-Safe Rechargeable Lithium-Ion Battery.

DTIC Science & Technology

1994-11-15

34 DEVELOPMENT OF AN ULTRA-SAFE RECHARGEABLE LITHIUM - ION BATTERY DTIC \\ JANI 0 1995 19941221 079 Contract # N00014-94-C-0141 ARPA Order...DEVELOPMENT OF AN ULTRA-SAFE RECHARGEABLE LITHIUM - ION BATTERY R&D STATUS REPORT 1931-1001/0 ARPA Order No.: 9332004arp01/13APR1994/313ES Program Code...Title of Work: Lithium - ion Battery Development Reporting Period: August 15, 1994 to November 15, 1994 Description of Progress: The project activities had

Legal Issues for the Commander,

DTIC Science & Technology

1983-01-01

Frederick T. Kiley, USAF Deputy Director, Plans and Programs Major William A. Buckingham, Jr., USAF Deputy Director, Administration Lieutenant (Junior...Writer-Editors Editorial Clerks Evelyn Lakes Pat Williams , Janis L. Hietala Lead Clerk Rebecca W. Miller Dorothy M. Mack Albert C. Helder Carol Valentine...9 M.J. 575 (A.F.C.M.R. 1980). 41 AL .. ~ ~. .~ ~ i 42 References 28. United States v. Poundstone , 22 U.S.C.M.A. 277, 46 C.M.R. 277 (1973). 29. United

Organizational Dysfunction in the US Air Force: Lessons from the ICBM Community

DTIC Science & Technology

2016-06-01

separated the space and missile career fields, highlighting a need to produce career professionals in the ICBM community.12 During this time, errors...raises for missileers,” Air Force Times 74, no. 29 (27 Jan, 2014): 10. 20 Gen Martin Dempsey, “America’s Military – A Profession of Arms.” Washington...Bullying”, 18-20; Cynthia Coccia, "Avoiding a "Toxic" Organization," Nursing Management 29, no. 5 (May 1998): 32; Janie Fritz. “Organizational Misbehavior

Mutagenesis and Characterization Studies to Develop Novel Bioluminescent Systems

DTIC Science & Technology

2010-05-12

described previously [42] and were corrected for the spectral response of the Turner TD -20e H6199 photomultiplier tube employed. b Tm, mean aggregation...the DNA encoding the biotin binding domain ( BBD , residues Met12-Val76 in the pET-KPBT-Luc plasmid) using the following primer and its respective...endonuclease sites are underlined). The BBD was amplified from the pET-KPBT-Luc plasmid by PCR (initial denaturation at 95 C for 2 min; a 60-cycle

Intraventricular Delivery of Engineered Oncolytic Herpes Simplex Virotherapy to Treat Localized and Metastatic Pediatric Brain Tumors

DTIC Science & Technology

2016-08-01

medulloblastoma (MB) to oHSV. Patient derived MB xenograft D341-luc, which is luciferase enabled, was used for the experiment. Athymic nude mice received...ability of G207 and M002 to kill another patient-derived pediatric group Page 10 3 MB xenograft D425-luc. Additionally, we will begin to test...from pediatric MB xenografts D425 and D341 (no identifiable information will be accessible to the research team by any means) and in Trp53-/- Ptch

Historical warming reduced due to enhanced land carbon uptake

PubMed Central

Shevliakova, Elena; Stouffer, Ronald J.; Malyshev, Sergey; Krasting, John P.; Hurtt, George C.; Pacala, Stephen W.

2013-01-01

Previous studies have demonstrated the importance of enhanced vegetation growth under future elevated atmospheric CO2 for 21st century climate warming. Surprisingly no study has completed an analogous assessment for the historical period, during which emissions of greenhouse gases increased rapidly and land-use changes (LUC) dramatically altered terrestrial carbon sources and sinks. Using the Geophysical Fluid Dynamics Laboratory comprehensive Earth System Model ESM2G and a reconstruction of the LUC, we estimate that enhanced vegetation growth has lowered the historical atmospheric CO2 concentration by 85 ppm, avoiding an additional 0.31 ± 0.06 °C warming. We demonstrate that without enhanced vegetation growth the total residual terrestrial carbon flux (i.e., the net land flux minus LUC flux) would be a source of 65–82 Gt of carbon (GtC) to atmosphere instead of the historical residual carbon sink of 186–192 GtC, a carbon saving of 251–274 GtC. PMID:24062452

Historical warming reduced due to enhanced land carbon uptake.

PubMed

Shevliakova, Elena; Stouffer, Ronald J; Malyshev, Sergey; Krasting, John P; Hurtt, George C; Pacala, Stephen W

2013-10-15

Previous studies have demonstrated the importance of enhanced vegetation growth under future elevated atmospheric CO2 for 21st century climate warming. Surprisingly no study has completed an analogous assessment for the historical period, during which emissions of greenhouse gases increased rapidly and land-use changes (LUC) dramatically altered terrestrial carbon sources and sinks. Using the Geophysical Fluid Dynamics Laboratory comprehensive Earth System Model ESM2G and a reconstruction of the LUC, we estimate that enhanced vegetation growth has lowered the historical atmospheric CO2 concentration by 85 ppm, avoiding an additional 0.31 ± 0.06 °C warming. We demonstrate that without enhanced vegetation growth the total residual terrestrial carbon flux (i.e., the net land flux minus LUC flux) would be a source of 65-82 Gt of carbon (GtC) to atmosphere instead of the historical residual carbon sink of 186-192 GtC, a carbon saving of 251-274 GtC.

Intracranial implantation with subsequent 3D in vivo bioluminescent imaging of murine gliomas.

PubMed

Abdelwahab, Mohammed G; Sankar, Tejas; Preul, Mark C; Scheck, Adrienne C

2011-11-06

The mouse glioma 261 (GL261) is recognized as an in vivo model system that recapitulates many of the features of human glioblastoma multiforme (GBM). The cell line was originally induced by intracranial injection of 3-methyl-cholantrene into a C57BL/6 syngeneic mouse strain (1); therefore, immunologically competent C57BL/6 mice can be used. While we use GL261, the following protocol can be used for the implantation and monitoring of any intracranial mouse tumor model. GL261 cells were engineered to stably express firefly luciferase (GL261-luc). We also created the brighter GL261-luc2 cell line by stable transfection of the luc2 gene expressed from the CMV promoter. C57BL/6-cBrd/cBrd/Cr mice (albino variant of C57BL/6) from the National Cancer Institute, Frederick, MD were used to eliminate the light attenuation caused by black skin and fur. With the use of albino C57BL/6 mice; in vivo imaging using the IVIS Spectrum in vivo imaging system is possible from the day of implantation (Caliper Life Sciences, Hopkinton, MA). The GL261-luc and GL261-luc2 cell lines showed the same in vivo behavior as the parental GL261 cells. Some of the shared histological features present in human GBMs and this mouse model include: tumor necrosis, pseudopalisades, neovascularization, invasion, hypercellularity, and inflammation (1). Prior to implantation animals were anesthetized by an intraperitoneal injection of ketamine (50 mg/kg), xylazine (5 mg/kg) and buprenorphine (0.05 mg/kg), placed in a stereotactic apparatus and an incision was made with a scalpel over the cranial midline. A burrhole was made 0.1 mm posterior to the bregma and 2.3mm to the right of the midline. A needle was inserted to a depth of 3mm and withdrawn 0.4 mm to a depth of 2.6 mm. Two μl of GL261-luc or GL261-luc2 cells (10(7) cells/ml) were infused over the course of 3 minutes. The burrhole was closed with bonewax and the incision was sutured. Following stereotactic implantation the bioluminescent cells are

[30 years since the first AIDS cases were reported: history and the present. Part I].

PubMed

Brůcková, Marie

2012-05-01

The 30-year natural history of AIDS disease is presented from the first clinical cases reported in 1981 to the identification of the HIV as the etiological agent of the disease. The priority dispute between Robert C. Gallo and Luc Montagnier over the discovery of the human immunodeficiency virus is briefly addressed. The final confirmation of the French priority was provided by the fact that the Nobel Prize in Medicine 2008 was awarded to Luc Montagnier and Francoise Barré--Sinoussi from the Pasteur Institute in Paris.

Directory of Researchers for Manpower, Personnel, Training, and Human Factors

DTIC Science & Technology

1998-03-01

Cambridge, MA Bohres, G Naval Surface Warfare Center - Dahlgren Div 904-234-4184 Panama City, FL DSN:436-4184 Bowers, Clint A Dr Univ of Central...Research 703-696-4318 Arlington, VA DSN:426-4318 Clevenger, J Evans and Sutherland Computer Corp 801-588-1518 Salt Lake City, UT Clingerman, B Space and...Drew W. CAPT Bergondy, M Bilazarian, P Bowers, Clint A Dr Bradley, S Breaux, R Mr Burke, J Burns, Christopher M Mr Burns, J Cannon-Bowers, Janis A Dr

A Handbook for Alcohol and Drug Control Officers. Volume I.

DTIC Science & Technology

1975-02-01

Lester R ., Management by Exception (New York: McGraw-Hill Book Co., 1964). *Cleland, David I ., Management (New York: McGraw-Hill Book Co., 1972). - * *Dale...b Option 1: By Battaliono n // Enough Matel ials I I . Yes ~Analyze objective to determine if a / Enough change in hours of training, number... 1971 ). IV-40 -4. i 16. R.A.J. Webb, "Fear and Communication," Journal of Drug Education 4:97-103 (Spring 1974). 17. A. A. Lumsdaine and I . L. Janis

Mutational Analysis of the Rift Valley Fever Virus Glycoprotein Precursor Proteins for Gn Protein Expression

PubMed Central

Phoenix, Inaia; Lokugamage, Nandadeva; Nishiyama, Shoko; Ikegami, Tetsuro

2016-01-01

The Rift Valley fever virus (RVFV) M-segment encodes the 78 kD, NSm, Gn, and Gc proteins. The 1st AUG generates the 78 kD-Gc precursor, the 2nd AUG generates the NSm-Gn-Gc precursor, and the 3rd AUG makes the NSm’-Gn-Gc precursor. To understand biological changes due to abolishment of the precursors, we quantitatively measured Gn secretion using a reporter assay, in which a Gaussia luciferase (gLuc) protein is fused to the RVFV M-segment pre-Gn region. Using the reporter assay, the relative expression of Gn/gLuc fusion proteins was analyzed among various AUG mutants. The reporter assay showed efficient secretion of Gn/gLuc protein from the precursor made from the 2nd AUG, while the removal of the untranslated region upstream of the 2nd AUG (AUG2-M) increased the secretion of the Gn/gLuc protein. Subsequently, recombinant MP-12 strains encoding mutations in the pre-Gn region were rescued, and virological phenotypes were characterized. Recombinant MP-12 encoding the AUG2-M mutation replicated slightly less efficiently than the control, indicating that viral replication is further influenced by the biological processes occurring after Gn expression, rather than the Gn abundance. This study showed that, not only the abolishment of AUG, but also the truncation of viral UTR, affects the expression of Gn protein by the RVFV M-segment. PMID:27231931

Reporter Gene-Facilitated Detection of Compounds in Arabidopsis Leaf Extracts that Activate the Karrikin Signaling Pathway.

PubMed

Sun, Yueming K; Flematti, Gavin R; Smith, Steven M; Waters, Mark T

2016-01-01

Karrikins are potent germination stimulants generated by the combustion of plant matter. Treatment of Arabidopsis with karrikins triggers a signaling process that is dependent upon a putative receptor protein KARRIKIN INSENSITIVE 2 (KAI2). KAI2 is a homolog of DWARF 14 (D14), the receptor for endogenous strigolactone hormones. Genetic analyses suggest that KAI2 also perceives endogenous signal(s) that are not strigolactones. Activation of KAI2 by addition of karrikins to Arabidopsis plants induces expression of transcripts including D14-LIKE 2 ( DLK2 ). We constructed the synthetic reporter gene DLK2 : LUC in Arabidopsis , which comprises the firefly luciferase gene ( LUC ) driven by the DLK2 promoter. Here we describe a luminescence-based reporter assay with Arabidopsis seeds to detect chemical signals that can activate the KAI2 signaling pathway. We demonstrate that the DLK2 : LUC assay can selectively and sensitively detect karrikins and a functionally similar synthetic strigolactone analog. Crucially we show that crude extracts from Arabidopsis leaves can also activate DLK2 : LUC in a KAI2-dependent manner. Our work provides the first direct evidence for the existence of endogenous chemical signals that can activate the KAI2-mediated signaling pathway in Arabidopsis . This sensitive reporter system can now be used for the bioassay-guided purification and identification of putative endogenous KAI2 ligands or their precursors, and endogenous compounds that might modulate the KAI2 signaling pathway.

Reporter Gene-Facilitated Detection of Compounds in Arabidopsis Leaf Extracts that Activate the Karrikin Signaling Pathway

PubMed Central

Sun, Yueming K.; Flematti, Gavin R.; Smith, Steven M.; Waters, Mark T.

2016-01-01

Karrikins are potent germination stimulants generated by the combustion of plant matter. Treatment of Arabidopsis with karrikins triggers a signaling process that is dependent upon a putative receptor protein KARRIKIN INSENSITIVE 2 (KAI2). KAI2 is a homolog of DWARF 14 (D14), the receptor for endogenous strigolactone hormones. Genetic analyses suggest that KAI2 also perceives endogenous signal(s) that are not strigolactones. Activation of KAI2 by addition of karrikins to Arabidopsis plants induces expression of transcripts including D14-LIKE 2 (DLK2). We constructed the synthetic reporter gene DLK2:LUC in Arabidopsis, which comprises the firefly luciferase gene (LUC) driven by the DLK2 promoter. Here we describe a luminescence-based reporter assay with Arabidopsis seeds to detect chemical signals that can activate the KAI2 signaling pathway. We demonstrate that the DLK2:LUC assay can selectively and sensitively detect karrikins and a functionally similar synthetic strigolactone analog. Crucially we show that crude extracts from Arabidopsis leaves can also activate DLK2:LUC in a KAI2-dependent manner. Our work provides the first direct evidence for the existence of endogenous chemical signals that can activate the KAI2-mediated signaling pathway in Arabidopsis. This sensitive reporter system can now be used for the bioassay-guided purification and identification of putative endogenous KAI2 ligands or their precursors, and endogenous compounds that might modulate the KAI2 signaling pathway. PMID:27994609

Developmental and Wound-, Cold-, Desiccation-, Ultraviolet-B-Stress-Induced Modulations in the Expression of the Petunia Zinc Finger Transcription Factor Gene ZPT2-21

PubMed Central

van der Krol, Alexander R.; van Poecke, Remco M.P.; Vorst, Oscar F.J.; Voogt, Charlotte; van Leeuwen, Wessel; Borst-Vrensen, Tanja W.M.; Takatsuji, Hiroshi; van der Plas, Linus H.W.

1999-01-01

The ZPT2-2 gene belongs to the EPF gene family in petunia (Petunia hybrida), which encodes proteins with TFIIIA-type zinc-finger DNA-binding motifs. To elucidate a possible function for ZPT2-2, we analyzed its pattern of expression in relation to different developmental and physiological stress signals. The activity of the ZPT2-2 promoter was analyzed using a firefly luciferase (LUC) reporter gene, allowing for continuous measurements of transgene activity in planta. We show that ZPT2-2::LUC is active in all plant tissues, but is strongly modulated in cotyledons upon germination, in leaves in response to desiccation, cold treatment, wounding, or ultraviolet-B light, and in petal tissue in response to pollination of the stigma. Analysis of mRNA levels indicated that the modulations in ZPT2-2::LUC expression reflect modulations in endogenous ZPT2-2 gene expression. The change in ZPT2-2::LUC activity by cold treatment, wounding, desiccation, and ultraviolet-B light suggest that the phytohormones ethylene and jasmonic acid are involved in regulating the expression of ZPT2-2. Although up-regulation of expression of ZPT2-2 can be blocked by inhibitors of ethylene perception, expression in plants is not induced by exogenously applied ethylene. The application of jasmonic acid does result in an up-regulation of gene activity and, thus, ZPT2-2 may play a role in the realization of the jasmonic acid hormonal responses in petunia. PMID:10594102

Climate extremes, land–climate feedbacks and land-use forcing at 1.5°C

DOE PAGES

Seneviratne, Sonia I.; Wartenburger, Richard; Guillod, Benoit P.; ...

2018-04-02

Here, this article investigates projected changes in temperature and water cycle extremes at 1.5°C of global warming, and highlights the role of land processes and land-use changes (LUCs) for these projections. We provide new comparisons of changes in climate at 1.5°C versus 2°C based on empirical sampling analyses of transient simulations versus simulations from the ‘Half a degree Additional warming, Prognosis and Projected Impacts’ (HAPPI) multi-model experiment. The two approaches yield similar overall results regarding changes in climate extremes on land, and reveal a substantial difference in the occurrence of regional extremes at 1.5°C versus 2°C. Land processes mediated throughmore » soil moisture feedbacks and land-use forcing play a major role for projected changes in extremes at 1.5°C in most mid-latitude regions, including densely populated areas in North America, Europe and Asia. This has important implications for low-emissions scenarios derived from integrated assessment models (IAMs), which include major LUCs in ambitious mitigation pathways (e.g. associated with increased bioenergy use), but are also shown to differ in the simulated LUC patterns. Biogeophysical effects from LUCs are not considered in the development of IAM scenarios, but play an important role for projected regional changes in climate extremes, and are thus of high relevance for sustainable development pathways.« less

Mutational Analysis of the Rift Valley Fever Virus Glycoprotein Precursor Proteins for Gn Protein Expression.

PubMed

Phoenix, Inaia; Lokugamage, Nandadeva; Nishiyama, Shoko; Ikegami, Tetsuro

2016-05-24

The Rift Valley fever virus (RVFV) M-segment encodes the 78 kD, NSm, Gn, and Gc proteins. The 1st AUG generates the 78 kD-Gc precursor, the 2nd AUG generates the NSm-Gn-Gc precursor, and the 3rd AUG makes the NSm'-Gn-Gc precursor. To understand biological changes due to abolishment of the precursors, we quantitatively measured Gn secretion using a reporter assay, in which a Gaussia luciferase (gLuc) protein is fused to the RVFV M-segment pre-Gn region. Using the reporter assay, the relative expression of Gn/gLuc fusion proteins was analyzed among various AUG mutants. The reporter assay showed efficient secretion of Gn/gLuc protein from the precursor made from the 2nd AUG, while the removal of the untranslated region upstream of the 2nd AUG (AUG2-M) increased the secretion of the Gn/gLuc protein. Subsequently, recombinant MP-12 strains encoding mutations in the pre-Gn region were rescued, and virological phenotypes were characterized. Recombinant MP-12 encoding the AUG2-M mutation replicated slightly less efficiently than the control, indicating that viral replication is further influenced by the biological processes occurring after Gn expression, rather than the Gn abundance. This study showed that, not only the abolishment of AUG, but also the truncation of viral UTR, affects the expression of Gn protein by the RVFV M-segment.

A dual function fusion protein of Herpes simplex virus type 1 thymidine kinase and firefly luciferase for noninvasive in vivo imaging of gene therapy in malignant glioma.

PubMed

Söling, Ariane; Theiss, Christian; Jungmichel, Stephanie; Rainov, Nikolai G

2004-08-04

BACKGROUND: Suicide gene therapy employing the prodrug activating system Herpes simplex virus type 1 thymidine kinase (HSV-TK)/ ganciclovir (GCV) has proven to be effective in killing experimental brain tumors. In contrast, glioma patients treated with HSV-TK/ GCV did not show significant treatment benefit, most likely due to insufficient transgene delivery to tumor cells. Therefore, this study aimed at developing a strategy for real-time noninvasive in vivo monitoring of the activity of a therapeutic gene in brain tumor cells. METHODS: The HSV-TK gene was fused to the firefly luciferase (Luc) gene and the fusion construct HSV-TK-Luc was expressed in U87MG human malignant glioma cells. Nude mice with subcutaneous gliomas stably expressing HSV-TK-Luc were subjected to GCV treatment and tumor response to therapy was monitored in vivo by serial bioluminescence imaging. Bioluminescent signals over time were compared with tumor volumes determined by caliper. RESULTS: Transient and stable expression of the HSV-TK-Luc fusion protein in U87MG glioma cells demonstrated close correlation of both enzyme activities. Serial optical imaging of tumor bearing mice detected in all cases GCV induced death of tumor cells expressing the fusion protein and proved that bioluminescence can be reliably used for repetitive and noninvasive quantification of HSV-TK/ GCV mediated cell kill in vivo. CONCLUSION: This approach may represent a valuable tool for the in vivo evaluation of gene therapy strategies for treatment of malignant disease.

3,3',5-Triiodo-L-thyronine-like activity in effluents from domestic sewage treatment plants detected by in vitro and in vivo bioassays

DOE Office of Scientific and Technical Information (OSTI.GOV)

Murata, Tomonori; Yamauchi, Kiyoshi

2008-02-01

Thyroid system-disrupting activity in effluents from municipal domestic sewage treatment plants was detected using three in vitro assays and one in vivo assay. Contaminants in the effluents were extracted by solid-phase extraction (SPE) and eluted stepwise with different organic solvents. The majority of the thyroid system-disrupting activity was detected in the dichloromethane/methanol (1/1) fraction after SPE in all three in vitro assays: competitive assays of 3,3',5-[{sup 125}I]triiodo-L-thyronine ([{sup 125}I]T{sub 3}) binding to the plasma protein transthyretin (TTR assay) and thyroid hormone receptor (TR assay) and T{sub 3}-dependent luciferase assay (Luc assay). Subsequent reverse-phase high-performance liquid chromatography (RP-HPLC) of the dichloromethane/methanolmore » (1/1) fraction separated contaminants potent in the TR and Luc assays from those potent in the TTR assay. The contaminants potent in the TR and Luc assays were also potent in an in vivo short-term gene expression assay in Xenopus laevis (Tadpole assay). The present study demonstrated that the effluents from domestic sewage treatment plants contain contaminants with T{sub 3}-like activity of {approx} 10{sup -10} M T{sub 3}-equivalent concentration (T{sub 3}EQ) and that the TR and Luc assays are powerful in vitro bioassays for detecting thyroid system-disrupting activity in effluents. The availability and applicability of these bioassays for screening contaminants with thyroid system-disrupting activity in the water environment are discussed.« less

Ultrasound-induced cavitation enhances the delivery and therapeutic efficacy of an oncolytic virus in an in vitro model.

PubMed

Bazan-Peregrino, Miriam; Arvanitis, Costas D; Rifai, Bassel; Seymour, Leonard W; Coussios, Constantin-C

2012-01-30

We investigated whether ultrasound-induced cavitation at 0.5 MHz could improve the extravasation and distribution of a potent breast cancer-selective oncolytic adenovirus, AdEHE2F-Luc, to tumour regions that are remote from blood vessels. We developed a novel tumour-mimicking model consisting of a gel matrix containing human breast cancer cells traversed by a fluid channel simulating a tumour blood vessel, through which the virus and microbubbles could be made to flow. Ultrasonic pressures were chosen to maximize either broadband emissions, associated with inertial cavitation, or ultraharmonic emissions, associated with stable cavitation, while varying duty cycle to keep the total acoustic energy delivered constant for comparison across exposures. None of the exposure conditions tested affected cell viability in the absence of the adenovirus. When AdEHE2F-Luc was delivered via the vessel, inertial cavitation increased transgene expression in tumour cells by up to 200 times. This increase was not observed in the absence of Coxsackie and Adenovirus Receptor cell expression, discounting sonoporation as the mechanism of action. In the presence of inertial cavitation, AdEHE2F-Luc distribution was greatly improved in the matrix surrounding the vessel, particularly in the direction of the ultrasound beam; this enabled AdEHE2F-Luc to kill up to 80% of cancer cells within the ultrasound focal volume in the gel 24 hours after delivery, compared to 0% in the absence of cavitation. Copyright © 2011 Elsevier B.V. All rights reserved.

Climate extremes, land–climate feedbacks and land-use forcing at 1.5°C

DOE Office of Scientific and Technical Information (OSTI.GOV)

Seneviratne, Sonia I.; Wartenburger, Richard; Guillod, Benoit P.

Here, this article investigates projected changes in temperature and water cycle extremes at 1.5°C of global warming, and highlights the role of land processes and land-use changes (LUCs) for these projections. We provide new comparisons of changes in climate at 1.5°C versus 2°C based on empirical sampling analyses of transient simulations versus simulations from the ‘Half a degree Additional warming, Prognosis and Projected Impacts’ (HAPPI) multi-model experiment. The two approaches yield similar overall results regarding changes in climate extremes on land, and reveal a substantial difference in the occurrence of regional extremes at 1.5°C versus 2°C. Land processes mediated throughmore » soil moisture feedbacks and land-use forcing play a major role for projected changes in extremes at 1.5°C in most mid-latitude regions, including densely populated areas in North America, Europe and Asia. This has important implications for low-emissions scenarios derived from integrated assessment models (IAMs), which include major LUCs in ambitious mitigation pathways (e.g. associated with increased bioenergy use), but are also shown to differ in the simulated LUC patterns. Biogeophysical effects from LUCs are not considered in the development of IAM scenarios, but play an important role for projected regional changes in climate extremes, and are thus of high relevance for sustainable development pathways.« less

Short Hairpin RNA Gene Silencing of Prolyl Hydroxylase-2 with a Minicircle Vector Improves Neovascularization of Hindlimb Ischemia

PubMed Central

Lijkwan, Maarten A.; Hellingman, Alwine A.; Bos, Ernst J.; van der Bogt, Koen E.A.; Huang, Mei; Kooreman, Nigel G.; de Vries, Margreet R.; Peters, Hendrika A.B.; Robbins, Robert C.; Quax, Paul H.A.

2014-01-01

Abstract In this study, we target the hypoxia inducible factor-1 alpha (HIF-1-alpha) pathway by short hairpin RNA interference therapy targeting prolyl hydroxylase-2 (shPHD2). We use the minicircle (MC) vector technology as an alternative for conventional nonviral plasmid (PL) vectors in order to improve neovascularization after unilateral hindlimb ischemia in a murine model. Gene expression and transfection efficiency of MC and PL, both in vitro and in vivo, were assessed using bioluminescence imaging (BLI) and firefly luciferase (Luc) reporter gene. C57Bl6 mice underwent unilateral electrocoagulation of the femoral artery and gastrocnemic muscle injection with MC-shPHD2, PL-shPHD2, or phosphate-buffered saline (PBS) as control. Blood flow recovery was monitored using laser Doppler perfusion imaging, and collaterals were visualized by immunohistochemistry and angiography. MC-Luc showed a 4.6-fold higher in vitro BLI signal compared with PL-Luc. BLI signals in vivo were 4.3×105±3.3×105 (MC-Luc) versus 0.4×105±0.3×105 (PL-Luc) at day 28 (p=0.016). Compared with PL-shPHD2 or PBS, MC-shPHD2 significantly improved blood flow recovery, up to 50% from day 3 until day 14 after ischemia induction. MC-shPHD2 significantly increased collateral density and capillary density, as monitored by alpha-smooth muscle actin expression and CD31+ expression, respectively. Angiography data confirmed the histological findings. Significant downregulation of PHD2 mRNA levels by MC-shPHD2 was confirmed by quantitative polymerase chain reaction. Finally, Western blot analysis confirmed significantly higher levels of HIF-1-alpha protein by MC-shPHD2, compared with PL-shPHD2 and PBS. This study provides initial evidence of a new potential therapeutic approach for peripheral artery disease. The combination of HIF-1-alpha pathway targeting by shPHD2 with the robust nonviral MC plasmid improved postischemic neovascularization, making this approach a promising potential treatment option for

Soil carbon, nitrogen and phosphorus changes under sugarcane expansion in Brazil.

PubMed

Franco, André L C; Cherubin, Maurício R; Pavinato, Paulo S; Cerri, Carlos E P; Six, Johan; Davies, Christian A; Cerri, Carlos C

2015-05-15

Historical data of land use change (LUC) indicated that the sugarcane expansion has mainly displaced pasture areas in Central-Southern Brazil, globally the largest producer, and that those pastures were prior established over native forests in the Cerrado biome. We sampled 3 chronosequences of land use comprising native vegetation (NV), pasture (PA), and sugarcane crop (SC) in the sugarcane expansion region to assess the effects of LUC on soil carbon, nitrogen, and labile phosphorus pools. Thirty years after conversion of NV to PA, we found significant losses of original soil organic matter (SOM) from NV, while insufficient new organic matter was introduced from tropical grasses into soil to offset the losses, reflecting in a net C emission of 0.4 Mg ha(-1)yr(-1). These findings added to decreases in (15)N signal indicated that labile portions of SOM are preserved under PA. Afterwards, in the firsts five years after LUC from PA to SC, sparse variations were found in SOM levels. After more than 20 years of sugarcane crop, however, there were losses of 40 and 35% of C and N stocks, respectively, resulting in a rate of C emission of 1.3 Mg ha(-1)yr(-1) totally caused by the respiration of SOM from C4-cycle plants. In addition, conversion of pastures to sugarcane mostly increased (15)N signal, indicating an accumulation of more recalcitrant SOM under sugarcane. The microbe- and plant-available P showed site-specific responses to LUC as a function of different P-input managements, with the biological pool mostly accounting for more than 50% of the labile P in both anthropic land uses. With the projections of 6.4 Mha of land required by 2021 for sugarcane expansion in Brazil to achieve ethanol's demand, this explanatory approach to the responses of SOM to LUC will contribute for an accurate assessment of the CO₂ balance of sugarcane ethanol. Copyright © 2015 Elsevier B.V. All rights reserved.

A non-invasive in vivo imaging system to study dissemination of bioluminescent Yersinia pestis CO92 in a mouse model of pneumonic plague.

PubMed

Sha, Jian; Rosenzweig, Jason A; Kirtley, Michelle L; van Lier, Christina J; Fitts, Eric C; Kozlova, Elena V; Erova, Tatiana E; Tiner, Bethany L; Chopra, Ashok K

2013-02-01

The gold standard in microbiology for monitoring bacterial dissemination in infected animals has always been viable plate counts. This method, despite being quantitative, requires sacrificing the infected animals. Recently, however, an alternative method of in vivo imaging of bioluminescent bacteria (IVIBB) for monitoring microbial dissemination within the host has been employed. Yersinia pestis is a Gram-negative bacterium capable of causing bubonic, septicemic, and pneumonic plague. In this study, we compared the conventional counting of bacterial colony forming units (cfu) in the various infected tissues to IVIBB in monitoring Y. pestis dissemination in a mouse model of pneumonic plague. By using a transposon mutagenesis system harboring the luciferase (luc) gene, we screened approximately 4000 clones and obtained a fully virulent, luc-positive Y. pestis CO92 (Y. pestis-luc2) reporter strain in which transposition occurred within the largest pMT1 plasmid which possesses murine toxin and capsular antigen encoding genes. The aforementioned reporter strain and the wild-type CO92 exhibited similar growth curves, formed capsule based on immunofluorescence microscopy and flow cytometry, and had a similar LD(50). Intranasal infection of mice with 15 LD(50) of CO92-luc2 resulted in animal mortality by 72 h, and an increasing number of bioluminescent bacteria were observed in various mouse organs over a 24-72 h period when whole animals were imaged. However, following levofloxacin treatment (10 mg/kg/day) for 6 days 24 h post infection, no luminescence was observed after 72 h of infection, indicating that the tested antimicrobial killed bacteria preventing their detection in host peripheral tissues. Overall, we demonstrated that IVIBB is an effective and non-invasive way of monitoring bacterial dissemination in animals following pneumonic plague having strong correlation with cfu, and our reporter CO92-luc2 strain can be employed as a useful tool to monitor the efficacy

The effect of surface demineralization of cortical bone allograft on the properties of recombinant adeno-associated virus coatings.

PubMed

Yazici, Cemal; Yanoso, Laura; Xie, Chao; Reynolds, David G; Samulski, R Jude; Samulski, Jade; Yannariello-Brown, Judith; Gertzman, Arthur A; Zhang, Xinping; Awad, Hani A; Schwarz, Edward M

2008-10-01

Freeze-dried recombinant adeno-associated virus (rAAV) coated structural allografts have emerged as an approach to engender necrotic cortical bone with host factors that will persist for weeks following surgery to facilitate revascularization, osteointegration, and remodeling. However, one major limitation is the nonporous cortical surface that prohibits uniform distribution of the rAAV coating prior to freeze-drying. To overcome this we have developed a demineralization method to increase surface absorbance while retaining the structural integrity of the allograft. Demineralized bone wafers (DBW) made from human femoral allograft rings demonstrated a significant 21.1% (73.6+/-3.9% versus 52.5+/-2.6%; p<0.001) increase in percent surface area coating versus mineralized controls. Co-incubation of rAAV-luciferase (rAAV-Luc) coated DBW with a monolayer of C3H10T1/2 cells in culture led to peak luciferase levels that were not significantly different from soluble rAAV-Luc controls (p>0.05), although the peaks occurred at 60h and 12h, respectively. To assess the transduction efficiency of rAAV-Luc coated DBW in vivo, we first performed a dose response with allografts containing 10(7), 10(9) or 10(10) particles that were surgically implanted into the quadriceps of mice, and assayed by in vivo bioluminescence imaging (BLI) on days 1, 3, 5, 7, 10, 14, and 21. The results demonstrated a dose response in which the DBW coated with 10(10) rAAV-Luc particles achieved peak gene expression levels on day 3, which persisted until day 21, and was significantly greater than the 10(7) dose throughout this time period (p<0.01). A direct comparison of mineralized versus DBW coated with 10(10) rAAV-Luc particles failed to demonstrate any significant differences in transduction kinetics or efficiency in vivo. Thus, surface demineralization of human cortical bone allograft increases its absorbance for uniform rAAV coating, without affecting vector transduction efficiency.

A toxin-antitoxin module as a target for antimicrobial development.

PubMed

Lioy, Virginia S; Rey, Oscar; Balsa, Dolors; Pellicer, Teresa; Alonso, Juan C

2010-01-01

The emergence and spread of pathogenic bacteria that have become resistant to multiple antibiotics through lateral gene transfer have created the need of novel antimicrobials. Toxin-antitoxin (TA) modules, which have been implicated in plasmid maintenance and stress management, are ubiquitous among plasmids from vancomycin or methicillin resistant bacteria. In the Streptococcus pyogenes pSM19035-encoded TA loci, the labile epsilon antitoxin binds to free zeta toxin and neutralizes it. When the zeta toxin is freed from the epsilon antitoxin, it induces a reversible state of growth arrest with a drastic reduction on the rate of replication, transcription and translation. However, upon prolonged zeta toxin action, the cells can no longer be rescued from their stasis state. A compound that disrupts the epsilon.zeta interaction can be considered as an attractive antimicrobial agent. Gene epsilon was fused to luc (Luc-epsilon antitoxin) and zeta to the gfp gene (zeta-GFP). Luc-epsilon or epsilon antitoxin neutralizes the toxic effect of the zeta or zeta-GFP toxin. In the absence of the antitoxin, free zeta or zeta-GFP triggers a reversible loss of cell proliferation, but the zetaK46A-GFP variant fails to block growth. Bioluminescence resonance energy transfer (BRET) assay was developed for high-throughput screening (HTS). To develop the proper controls, molecular dynamics studies were used to predict that the Asp18 and/or Glu22 residues might be relevant for epsilon.zeta interaction. Luc-epsilon efficiently transfers the excited energy to the fluorescent acceptor molecule (zeta-GFP or zetaK46A-GFP) and rendered high bioluminescence BRET signals. The exchange of Asp18 to Ala from zeta (D18A) affects Luc-epsilon.zetaD18A K46A-GFP interaction. In this study, we validate the hypothesis that it is possible to disrupt a TA module and offer a novel and unexploited targets to fight against antibiotic-resistant strains. Copyright 2009 Elsevier Inc. All rights reserved.

A non-invasive in vivo imaging system to study dissemination of bioluminescent Yersinia pestis CO92 in a mouse model of pneumonic plague

PubMed Central

Sha, Jian; Rosenzweig, Jason A.; Kirtley, Michelle L.; van Lier, Christina J.; Fitts, Eric C.; Kozlova, Elena V.; Erova, Tatiana E.; Tiner, Bethany L.; Chopra, Ashok K.

2012-01-01

The gold standard in microbiology for monitoring bacterial dissemination in infected animals has always been viable plate counts. This method, despite being quantitative, requires sacrificing the infected animals. Recently, however, an alternative method of in vivo imaging of bioluminescent bacteria (IVIBB) for monitoring microbial dissemination within the host has been employed. Yersina pestis is a Gram-negative bacterium capable of causing bubonic, septicemic, and pneumonic plague. In this study, we compared the conventional counting of bacterial colony forming units (cfu) in the various infected tissues to IVIBB in monitoring Y. pestis dissemination in a mouse model of pneumonic plague. By using a transposon mutagenesis system harboring the luciferase (luc) gene, we screened approximately 4000 clones and obtained a fully virulent, luc-positive Y. pestis CO92 (Y. pestis-luc2) reporter strain in which transposition occurred within the largest pMT1 plasmid which possesses murine toxin and capsular antigen encoding genes. The aforementioned reporter strain and the wild-type CO92 exhibited similar growth curves, formed capsule based on immunofluorescence microscopy and flow cytometry, and had a similar LD50. Intranasal infection of mice with 15 LD50 of CO92-luc2 resulted in animal mortality by 72 h, and an increasing number of bioluminescent bacteria were observed in various mouse organs over a 24–72 h period when whole animals were imaged. However, following levofloxacin treatment (10 mg/kg/day) for 6 days 24 h post infection, no luminescence was observed after 72 h of infection, indicating that the tested antimicrobial killed bacteria preventing their detection in host peripheral tissues. Overall, we demonstrated that IVIBB is an effective and non-invasive way of monitoring bacterial dissemination in animals following pneumonic plague having strong correlation with cfu, and our reporter CO92-luc2 strain can be employed as a useful tool to monitor the efficacy of

Loss of soil (macro)fauna due to the expansion of Brazilian sugarcane acreage.

PubMed

Franco, André L C; Bartz, Marie L C; Cherubin, Maurício R; Baretta, Dilmar; Cerri, Carlos E P; Feigl, Brigitte J; Wall, Diana H; Davies, Christian A; Cerri, Carlos C

2016-09-01

Land use changes (LUC) from pasture to sugarcane (Saccharum spp.) crop are expected to add 6.4Mha of new sugarcane land by 2021 in the Brazilian Cerrado and Atlantic Forest biomes. We assessed the effects of these LUC on the abundance and community structure of animals that inhabit soils belowground through a field survey using chronosequences of land uses comprising native vegetation, pasture, and sugarcane along a 1000-km-long transect across these two major tropical biomes in Brazil. Macrofauna community composition differed among land uses. While most groups were associated with samples taken in native vegetation, high abundance of termites and earthworms appeared associated with pasture soils. Linear mixed effects analysis showed that LUC affected total abundance (X(2)(1)=6.79, p=0.03) and taxa richness (X(2)(1)=6.08, p=0.04) of soil macrofauna. Abundance increased from 411±70individualsm(-2) in native vegetation to 1111±202individualsm(-2) in pasture, but decreased sharply to 106±24individualsm(-2) in sugarcane soils. Diversity decreased 24% from native vegetation to pasture, and 39% from pasture to sugarcane. Thus, a reduction of ~90% in soil macrofauna abundance, besides a loss of ~40% in the diversity of macrofauna groups, can be expected when sugarcane crops replace pasture in Brazilian tropical soils. In general, higher abundances of major macrofauna groups (ants, coleopterans, earthworms, and termites) were associated with higher acidity and low contents of macronutrients and organic matter in soil. This study draws attention for a significant biodiversity loss belowground due to tropical LUC in sugarcane expansion areas. Given that many groups of soil macrofauna are recognized as key mediators of ecosystem processes such as soil aggregation, nutrients cycling and soil carbon storage, our results warrant further efforts to understand the impacts of altering belowground biodiversity and composition on soil functioning and agriculture performance

Amelioration of radiation-induced pulmonary fibrosis by a water-soluble bifunctional sulfoxide radiation mitigator (MMS350).

PubMed

Kalash, Ronny; Epperly, Michael W; Goff, Julie; Dixon, Tracy; Sprachman, Melissa M; Zhang, Xichen; Shields, Donna; Cao, Shaonan; Franicola, Darcy; Wipf, Peter; Berhane, Hebist; Wang, Hong; Au, Jeremiah; Greenberger, Joel S

2013-11-01

A water-soluble ionizing radiation mitigator would have considerable advantages for the management of acute and chronic effects of ionizing radiation. We report that a novel oxetanyl sulfoxide (MMS350) is effective both as a protector and a mitigator of clonal mouse bone marrow stromal cell lines in vitro, and is an effective in vivo mitigator when administered 24 h after 9.5 Gy (LD100/30) total-body irradiation of C57BL/6NHsd mice, significantly improving survival (P = 0.0097). Furthermore, MMS350 (400 μM) added weekly to drinking water after 20 Gy thoracic irradiation significantly decreased: expression of pulmonary inflammatory and profibrotic gene transcripts and proteins; migration into the lungs of bone marrow origin luciferase+/GFP+ (luc+/GFP+) fibroblast progenitors (in both luc+ marrow chimeric and luc+ stromal cell line injected mouse models) and decreased radiation-induced pulmonary fibrosis (P < 0.0001). This nontoxic and orally administered small molecule may be an effective therapeutic in clinical radiotherapy and as a counter measure against the acute and chronic effects of ionizing radiation.

Kennedy Space Center Press Site (SWMU 074) Interim Measure Report

NASA Technical Reports Server (NTRS)

Applegate, Joseph L.

2015-01-01

This report summarizes the Interim Measure (IM) activities conducted at the Kennedy Space Center (KSC) Press Site ("the Press Site"). This facility has been designated as Solid Waste Management Unit 074 under KSC's Resource Conservation and Recovery Act Corrective Action program. The activities were completed as part of the Vehicle Assembly Building (VAB) Area Land Use Controls Implementation Plan (LUCIP) Elimination Project. The purpose of the VAB Area LUCIP Elimination Project was to delineate and remove soil affected with constituents of concern (COCs) that historically resulted in Land Use Controls (LUCs). The goal of the project was to eliminate the LUCs on soil. LUCs for groundwater were not addressed as part of the project and are not discussed in this report. This report is intended to meet the Florida Department of Environmental Protection (FDEP) Corrective Action Management Plan requirement as part of the KSC Hazardous and Solid Waste Amendments permit and the U.S. Environmental Protection Agency's (USEPA's) Toxic Substance Control Act (TSCA) self-implementing polychlorinated biphenyl (PCB) cleanup requirements of 40 Code of Federal Regulations (CFR) 761.61(a).

Amelioration of Radiation-Induced Pulmonary Fibrosis by a Water-Soluble Bifunctional Sulfoxide Radiation Mitigator (MMS350)

PubMed Central

Kalash, Ronny; Epperly, Michael W.; Goff, Julie; Dixon, Tracy; Sprachman, Melissa M.; Zhang, Xichen; Shields, Donna; Cao, Shaonan; Franicola, Darcy; Wipf, Peter; Berhane, Hebist; Wang, Hong; Au, Jeremiah; Greenberger, Joel S.

2014-01-01

A water-soluble ionizing radiation mitigator would have considerable advantages for the management of acute and chronic effects of ionizing radiation. We report that a novel oxetanyl sulfoxide (MMS350) is effective both as a protector and a mitigator of clonal mouse bone marrow stromal cell lines in vitro, and is an effective in vivo mitigator when administered 24 h after 9.5 Gy (LD100/30) total-body irradiation of C57BL/6NHsd mice, significantly improving survival (P =0.0097). Furthermore, MMS350 (400 μM) added weekly to drinking water after 20 Gy thoracic irradiation significantly decreased: expression of pulmonary inflammatory and profibrotic gene transcripts and proteins; migration into the lungs of bone marrow origin luciferase+/GFP+ (luc+/GFP+) fibroblast progenitors (in both luc+ marrow chimeric and luc+ stromal cell line injected mouse models) and decreased radiation-induced pulmonary fibrosis (P < 0.0001). This nontoxic and orally administered small molecule may be an effective therapeutic in clinical radiotherapy and as a counter measure against the acute and chronic effects of ionizing radiation. PMID:24125487

Novel Locally Active Estrogens Accelerate Cutaneous Wound Healing-Part 2.

PubMed

Brufani, Mario; Rizzi, Nicoletta; Meda, Clara; Filocamo, Luigi; Ceccacci, Francesca; D'Aiuto, Virginia; Bartoli, Gabriele; Bella, Angela La; Migneco, Luisa M; Bettolo, Rinaldo Marini; Leonelli, Francesca; Ciana, Paolo; Maggi, Adriana

2017-05-31

Estrogen deprivation is associated with delayed healing, while estrogen replacement therapy (ERT) accelerates acute wound healing and protects against development of chronic wounds. However, current estrogenic molecules have undesired systemic effects, thus the aim of our studies is to generate new molecules for topic administration that are devoid of systemic effects. Following a preliminary study, the new 17β-estradiol derivatives 1 were synthesized. The estrogenic activity of these novel compounds was evaluated in vitro using the cell line ERE-Luc B17 stably transfected with an ERE-Luc reporter. Among the 17β-estradiol derivatives synthesized, compounds 1e and 1f showed the highest transactivation potency and were therefore selected for the study of their systemic estrogenic activity. The study of these compounds in the ERE-Luc mouse model demonstrated that both compounds lack systemic effects when administered in the wound area. Furthermore, wound-healing experiments showed that 1e displays a significant regenerative and anti-inflammatory activity. It is therefore confirmed that this class of compounds are suitable for topical administration and have a clear beneficial effect on wound healing.

Cinnamic acid shortens the period of the circadian clock in mice.

PubMed

Oishi, Katsutaka; Yamamoto, Saori; Oike, Hideaki; Ohkura, Naoki; Taniguchi, Masahiko

2017-03-01

Cinnamic acid (CA) derivatives have recently received focus due to their anticancer, antioxidant, and antidiabetic properties. The present study aimed to determine the effects of cinnamic acid on the circadian clock, which is a cell-autonomous endogenous system that generates circadian rhythms that govern the behavior and physiology of most organisms. Cinnamic acid significantly shortened the circadian period of PER2::LUC expression in neuronal cells that differentiated from neuronal progenitor cells derived from PER2::LUC mouse embryos. Cinnamic acid did not induce the transient mRNA expression of clock genes such as Per1 and Per2 in neuronal cells, but significantly shortened the half-life of PER2::LUC protein in neuronal cells incubated with actinomycin D, suggested that CA post-transcriptionally affects the molecular clock by decreasing Per2 mRNA stability. A continuous infusion of CA into mice via an Alzet osmotic pump under constant darkness significantly shortened the free-running period of wheel-running rhythms. These findings suggest that CA shortens the circadian period of the molecular clock in mammals.

Land-Use Change and Emerging Infectious Disease on an Island Continent

PubMed Central

McFarlane, Rosemary A.; Sleigh, Adrian C.; McMichael, Anthony J.

2013-01-01

A more rigorous and nuanced understanding of land-use change (LUC) as a driver of emerging infectious disease (EID) is required. Here we examine post hunter-gatherer LUC as a driver of infectious disease in one biogeographical region with a compressed and documented history—continental Australia. We do this by examining land-use and native vegetation change (LUCC) associations with infectious disease emergence identified through a systematic (1973–2010) and historical (1788–1973) review of infectious disease literature of humans and animals. We find that 22% (20) of the systematically reviewed EIDs are associated with LUCC, most frequently where natural landscapes have been removed or replaced with agriculture, plantations, livestock or urban development. Historical clustering of vector-borne, zoonotic and environmental disease emergence also follows major periods of extensive land clearing. These advanced stages of LUCC are accompanied by changes in the distribution and density of hosts and vectors, at varying scales and chronology. This review of infectious disease emergence in one continent provides valuable insight into the association between accelerated global LUC and concurrent accelerated infectious disease emergence. PMID:23812027

Climate extremes, land–climate feedbacks and land-use forcing at 1.5°C

PubMed Central

Wartenburger, Richard; Guillod, Benoit P.; Hirsch, Annette L.; Vogel, Martha M.; Brovkin, Victor; van Vuuren, Detlef P.; Schaller, Nathalie; Boysen, Lena; Calvin, Katherine V.; Doelman, Jonathan; Greve, Peter; Havlik, Petr; Humpenöder, Florian; Krisztin, Tamas; Mitchell, Daniel; Popp, Alexander; Riahi, Keywan; Rogelj, Joeri; Schleussner, Carl-Friedrich; Sillmann, Jana; Stehfest, Elke

2018-01-01

This article investigates projected changes in temperature and water cycle extremes at 1.5°C of global warming, and highlights the role of land processes and land-use changes (LUCs) for these projections. We provide new comparisons of changes in climate at 1.5°C versus 2°C based on empirical sampling analyses of transient simulations versus simulations from the ‘Half a degree Additional warming, Prognosis and Projected Impacts’ (HAPPI) multi-model experiment. The two approaches yield similar overall results regarding changes in climate extremes on land, and reveal a substantial difference in the occurrence of regional extremes at 1.5°C versus 2°C. Land processes mediated through soil moisture feedbacks and land-use forcing play a major role for projected changes in extremes at 1.5°C in most mid-latitude regions, including densely populated areas in North America, Europe and Asia. This has important implications for low-emissions scenarios derived from integrated assessment models (IAMs), which include major LUCs in ambitious mitigation pathways (e.g. associated with increased bioenergy use), but are also shown to differ in the simulated LUC patterns. Biogeophysical effects from LUCs are not considered in the development of IAM scenarios, but play an important role for projected regional changes in climate extremes, and are thus of high relevance for sustainable development pathways. This article is part of the theme issue ‘The Paris Agreement: understanding the physical and social challenges for a warming world of 1.5°C above pre-industrial levels'. PMID:29610382

Effects of inorganic polyphosphate on bone sialoprotein gene expression.

PubMed

Wang, Zhitao; Li, Xinyue; Li, Zhengyang; Yang, Li; Sasaki, Yoko; Wang, Shuang; Zhou, Liming; Araki, Shouta; Mezawa, Masaru; Takai, Hideki; Ogata, Yorimasa

2010-03-01

Inorganic polyphosphate (poly(P)) is a biopolymer existing in almost all cells and tissues. The biological functions of poly(P) in micro-organisms have been extensively investigated in studies of poly(P) in eukaryotic cells, especially osteoblasts, and are increasing. Bone sialoprotein (BSP) is thought to function in bone mineralization, and is selectively expressed by differentiated osteoblasts. In this study, application of sodium phosphate glass type 25 (SPG25, 12.5 and 125 microM) increased BSP mRNA levels at 12 h in osteoblast-like ROS 17/2.8 cells. In transient transfection assay, 12.5 and 125 microM SPG25 increased luciferase activities of the constructs pLUC3 (-116 to +60), pLUC4 (-425 to +60), pLUC5 (-801 to +60) and pLUC6 (-938 to +60). Introduction of 2 bp mutations to the luciferase constructs showed that the effects of SPG25 were mediated by a FGF2 response element (FRE) and a homeodomain protein binding site (HOX). Luciferase activities induced by SPG25 were blocked by tyrosine kinase inhibitor herbimycine A, MAP kinase kinase inhibitor U0126, PI3-kinase/Akt inhibitor LY249002 and inorganic phosphate transport inhibitor foscarnet. Gel shift analyses showed that both 12.5 and 125 microM SPG25 increased nuclear protein binding to FRE and HOX elements. These studies demonstrate that SPG25 stimulates BSP transcription by targeting FRE and HOX elements in the proximal promoter of the rat BSP gene. Copyright 2010 Elsevier B.V. All rights reserved.

Climate extremes, land-climate feedbacks and land-use forcing at 1.5°C.

PubMed

Seneviratne, Sonia I; Wartenburger, Richard; Guillod, Benoit P; Hirsch, Annette L; Vogel, Martha M; Brovkin, Victor; van Vuuren, Detlef P; Schaller, Nathalie; Boysen, Lena; Calvin, Katherine V; Doelman, Jonathan; Greve, Peter; Havlik, Petr; Humpenöder, Florian; Krisztin, Tamas; Mitchell, Daniel; Popp, Alexander; Riahi, Keywan; Rogelj, Joeri; Schleussner, Carl-Friedrich; Sillmann, Jana; Stehfest, Elke

2018-05-13

This article investigates projected changes in temperature and water cycle extremes at 1.5°C of global warming, and highlights the role of land processes and land-use changes (LUCs) for these projections. We provide new comparisons of changes in climate at 1.5°C versus 2°C based on empirical sampling analyses of transient simulations versus simulations from the 'Half a degree Additional warming, Prognosis and Projected Impacts' (HAPPI) multi-model experiment. The two approaches yield similar overall results regarding changes in climate extremes on land, and reveal a substantial difference in the occurrence of regional extremes at 1.5°C versus 2°C. Land processes mediated through soil moisture feedbacks and land-use forcing play a major role for projected changes in extremes at 1.5°C in most mid-latitude regions, including densely populated areas in North America, Europe and Asia. This has important implications for low-emissions scenarios derived from integrated assessment models (IAMs), which include major LUCs in ambitious mitigation pathways (e.g. associated with increased bioenergy use), but are also shown to differ in the simulated LUC patterns. Biogeophysical effects from LUCs are not considered in the development of IAM scenarios, but play an important role for projected regional changes in climate extremes, and are thus of high relevance for sustainable development pathways.This article is part of the theme issue 'The Paris Agreement: understanding the physical and social challenges for a warming world of 1.5°C above pre-industrial levels'. © 2018 The Authors.

Adenovirus vector covalently conjugated to polyethylene glycol with a cancer-specific promoter suppresses the tumor growth through systemic administration.

PubMed

Yao, Xinglei; Yoshioka, Yasuo; Morishige, Tomohiro; Eto, Yusuke; Narimatsu, Shogo; Mizuguchi, Hiroyuki; Mukai, Yohei; Okada, Naoki; Nakagawa, Shinsaku

2010-01-01

Cancer gene therapy with adenovirus vectors (Adv) is limited to local administration because systemic administration of Adv produces a weak therapeutic effect and severe side effects. Previously, we generated a dual cancer-specific Adv system by using Adv covalently conjugated to polyethylene glycol (PEG) for transductional targeting and the telomere reverse transcriptase (TERT) promoter as a cancer-specific promoter for transcriptional targeting (PEG-Ad-TERT). We demonstrated that systemic administration of PEG-Ad-TERT showed superior antitumor effects against lung metastatic cancer with negligible side effects. Here, we investigated the therapeutic efficacy of systemic administration of PEG-Ad-TERT for the treatment of primary tumors. We first evaluated the transgene expression of PEG-Ad-TERT containing the luciferase gene (PEG-Ad-TERT/Luc) in primary tumors. Systemic administration of PEG-Ad-TERT/Luc resulted high transgene expression, similar to that observed in tumors for the conventional cytomegalovirus (CMV) promoter-driven Adv containing the luciferase gene (Ad-CMV/Luc). By comparison, transgene expression was 2500-fold lower than that of Ad-CMV/Luc in liver. We then examined the therapeutic effect of systemic administration of PEG-Ad-TERT containing the herpes simplex virus thymidine kinase (HSVtk) gene (PEG-Ad-TERT/HSVtk) for the treatment of primary tumors. We showed that PEG-Ad-TERT/HSVtk produced a notable antitumor effect against primary tumors with negligible side effects. These results demonstrated that PEG-Ad-TERT can be regarded as a prototype Adv with suitable efficacy and safety for systemic cancer gene therapy against both metastatic and primary tumors.

The Biosphere Under Potential Paris Outcomes

NASA Astrophysics Data System (ADS)

Ostberg, Sebastian; Boysen, Lena R.; Schaphoff, Sibyll; Lucht, Wolfgang; Gerten, Dieter

2018-01-01

Rapid economic and population growth over the last centuries have started to push the Earth out of its Holocene state into the Anthropocene. In this new era, ecosystems across the globe face mounting dual pressure from human land use change (LUC) and climate change (CC). With the Paris Agreement, the international community has committed to holding global warming below 2°C above preindustrial levels, yet current pledges by countries to reduce greenhouse gas emissions appear insufficient to achieve that goal. At the same time, the sustainable development goals strive to reduce inequalities between countries and provide sufficient food, feed, and clean energy to a growing world population likely to reach more than 9 billion by 2050. Here, we present a macro-scale analysis of the projected impacts of both CC and LUC on the terrestrial biosphere over the 21st century using the Representative Concentration Pathways (RCPs) to illustrate possible trajectories following the Paris Agreement. We find that CC may cause major impacts in landscapes covering between 16% and 65% of the global ice-free land surface by the end of the century, depending on the success or failure of achieving the Paris goal. Accounting for LUC impacts in addition, this number increases to 38%-80%. Thus, CC will likely replace LUC as the major driver of ecosystem change unless global warming can be limited to well below 2°C. We also find a substantial risk that impacts of agricultural expansion may offset some of the benefits of ambitious climate protection for ecosystems.

Climate extremes, land-climate feedbacks and land-use forcing at 1.5°C

NASA Astrophysics Data System (ADS)

Seneviratne, Sonia I.; Wartenburger, Richard; Guillod, Benoit P.; Hirsch, Annette L.; Vogel, Martha M.; Brovkin, Victor; van Vuuren, Detlef P.; Schaller, Nathalie; Boysen, Lena; Calvin, Katherine V.; Doelman, Jonathan; Greve, Peter; Havlik, Petr; Humpenöder, Florian; Krisztin, Tamas; Mitchell, Daniel; Popp, Alexander; Riahi, Keywan; Rogelj, Joeri; Schleussner, Carl-Friedrich; Sillmann, Jana; Stehfest, Elke

2018-05-01

This article investigates projected changes in temperature and water cycle extremes at 1.5°C of global warming, and highlights the role of land processes and land-use changes (LUCs) for these projections. We provide new comparisons of changes in climate at 1.5°C versus 2°C based on empirical sampling analyses of transient simulations versus simulations from the `Half a degree Additional warming, Prognosis and Projected Impacts' (HAPPI) multi-model experiment. The two approaches yield similar overall results regarding changes in climate extremes on land, and reveal a substantial difference in the occurrence of regional extremes at 1.5°C versus 2°C. Land processes mediated through soil moisture feedbacks and land-use forcing play a major role for projected changes in extremes at 1.5°C in most mid-latitude regions, including densely populated areas in North America, Europe and Asia. This has important implications for low-emissions scenarios derived from integrated assessment models (IAMs), which include major LUCs in ambitious mitigation pathways (e.g. associated with increased bioenergy use), but are also shown to differ in the simulated LUC patterns. Biogeophysical effects from LUCs are not considered in the development of IAM scenarios, but play an important role for projected regional changes in climate extremes, and are thus of high relevance for sustainable development pathways. This article is part of the theme issue `The Paris Agreement: understanding the physical and social challenges for a warming world of 1.5°C above pre-industrial levels'.

Time delay and magnification centroid due to gravitational lensing by black holes and naked singularities

DOE Office of Scientific and Technical Information (OSTI.GOV)

Virbhadra, K. S.; Keeton, C. R.; Department of Physics and Astronomy, Rutgers University, 136 Frelinghuysen Road, Piscataway, NJ 08854

We model the massive dark object at the center of the Galaxy as a Schwarzschild black hole as well as Janis-Newman-Winicour naked singularities, characterized by the mass and scalar charge parameters, and study gravitational lensing (particularly time delay, magnification centroid, and total magnification) by them. We find that the lensing features are qualitatively similar (though quantitatively different) for Schwarzschild black holes, weakly naked, and marginally strongly naked singularities. However, the lensing characteristics of strongly naked singularities are qualitatively very different from those due to Schwarzschild black holes. The images produced by Schwarzschild black hole lenses and weakly naked and marginallymore » strongly naked singularity lenses always have positive time delays. On the other hand, strongly naked singularity lenses can give rise to images with positive, zero, or negative time delays. In particular, for a large angular source position the direct image (the outermost image on the same side as the source) due to strongly naked singularity lensing always has a negative time delay. We also found that the scalar field decreases the time delay and increases the total magnification of images; this result could have important implications for cosmology. As the Janis-Newman-Winicour metric also describes the exterior gravitational field of a scalar star, naked singularities as well as scalar star lenses, if these exist in nature, will serve as more efficient cosmic telescopes than regular gravitational lenses.« less

Land Use Control Implementation Plan

NASA Technical Reports Server (NTRS)

Starr, Andrew Scott

2015-01-01

This Land Use Control Implementation Plan (LUCIP) has been prepared to inform current and potential future users of Building M7-505 of institutional controls that have been implemented at the site. Although there are no current unacceptable risks to human health or the environment associated with Building M7-505, institutional land use controls (LUCs) are necessary to prohibit the use of groundwater from the site. LUCs are also necessary to prevent access to soil under electrical equipment in the northwest portion of the site. Controls necessary to prevent human exposure will include periodic inspection, condition certification, and agency notification.

FGF-23 Regulates CYP27B1 Transcription in the Kidney and in Extra-Renal Tissues

PubMed Central

Chanakul, Ankanee; Zhang, Martin Y. H.; Louw, Andrew; Armbrecht, Harvey J.; Miller, Walter L.; Portale, Anthony A.; Perwad, Farzana

2013-01-01

The mitochondrial enzyme 25-hydroxyvitamin D 1α-hydroxylase, which is encoded by the CYP27B1 gene, converts 25OHD to the biological active form of vitamin D, 1,25-dihydroxyvitamin D (1,25(OH)2D). Renal 1α-hydroxylase activity is the principal determinant of the circulating 1,25(OH)2D concentration and enzyme activity is tightly regulated by several factors. Fibroblast growth factor-23 (FGF-23) decreases serum 1,25(OH)2D concentrations by suppressing CYP27B1 mRNA abundance in mice. In extra-renal tissues, 1α-hydroxylase is responsible for local 1,25(OH)2D synthesis, which has important paracrine actions, but whether FGF-23 regulates CYP27B1 gene expression in extra-renal tissues is unknown. We sought to determine whether FGF-23 regulates CYP27B1 transcription in the kidney and whether extra-renal tissues are target sites for FGF-23-induced suppression of CYP27B1. In HEK293 cells transfected with the human CYP27B1 promoter, FGF-23 suppressed promoter activity by 70%, and the suppressive effect was blocked by CI-1040, a specific inhibitor of extracellular signal regulated kinase 1/2. To examine CYP27B1 transcriptional activity in vivo, we crossed fgf-23 null mice with mice bearing the CYP27B1 promoter-driven luciferase transgene (1α-Luc). In the kidney of FGF-23 null/1α-Luc mice, CYP27B1 promoter activity was increased by 3-fold compared to that in wild-type/1α-Luc mice. Intraperitoneal injection of FGF-23 suppressed renal CYP27B1 promoter activity and protein expression by 26% and 60% respectively, and the suppressive effect was blocked by PD0325901, an ERK1/2 inhibitor. These findings provide evidence that FGF-23 suppresses CYP27B1 transcription in the kidney. Furthermore, we demonstrate that in FGF-23 null/1α-Luc mice, CYP27B1 promoter activity and mRNA abundance are increased in several extra-renal sites. In the heart of FGF-23 null/1α-Luc mice, CYP27B1 promoter activity and mRNA were 2- and 5-fold higher, respectively, than in control mice. We also

Continuation of Novel Analytical and Empircal Approaches to the Origin and Prediction of Pathogenicity

DTIC Science & Technology

2014-01-31

properties of strains as encoded by key mutations (e.g., E627K in PB2 in H5N1 or H7N9 , which confers increased replication in mammals) (Janies et al...the recent outbreak of H7N9 in China. We have created SUPRAMAPs and ROUTEMAPs describing the host and geographic origins and movement of each...segment. Here is a summary of our results:  The China-Taiwan H7N9 outbreak in early 2013 was caused by a reassortant virus made primarily of genetic

Geodesic motion around traversable wormholes supported by a massless conformally coupled scalar field

NASA Astrophysics Data System (ADS)

Willenborg, Felix; Grunau, Saskia; Kleihaus, Burkhard; Kunz, Jutta

2018-06-01

We consider a traversable wormhole solution of Einstein's gravity conformally coupled to a massless scalar field, a solution derived by Barcelo and Visser based on the Janis-Newman-Winicour-Wyman spacetime. We study the geodesic motion of timelike and spacelike particles in this spacetime. We solve the equations of motion analytically in terms of the Weierstraß functions and discuss all possible orbit types and their parameter dependence. Interestingly, bound orbits occur for timelike geodesics only in one of the two worlds. Moreover, under no conditions there exist timelike two world bound orbits.

Development of a luciferase based viral inhibition assay to evaluate vaccine induced CD8 T-cell responses

PubMed Central

Naarding, Marloes A.; Fernandez-Fernandez, Natalia; Kappes, John C.; Hayes, Peter; Ahmed, Tina; Icyuz, Mert; Edmonds, Tara G.; Bergin, Philip; Anzala, Omu; Hanke, Tomas; Clark, Lorna; Cox, Josephine H.; Cormier, Emmanuel; Ochsenbauer, Christina; Gilmour, Jill

2014-01-01

Emergence of SIV and HIV specific CD8 T cells has been shown to correlate with control of in vivo replication. Poor correlation between IFN-γ ELISPOT responses and in vivo control of the virus has triggered the development of more relevant assays to assess functional HIV-1 specific CD8 T-cell responses for the evaluation and prioritization of new HIV-1 vaccine candidates. We previously established a viral inhibition assay (VIA) that measures the ability of vaccine-induced CD8 T-cell responses to inhibit viral replication in autologous CD4 T cells. In this assay, viral replication is determined by measuring p24 in the culture supernatant. Here we describe the development of a novel VIA, referred to as IMC LucR VIA that exploits replication-competent HIV-1 infectious molecular clones (IMCs) in which the complete proviral genome is strain-specific and which express the Renilla luciferase (LucR) gene to determine viral growth and inhibition. The introduction of the luciferase readout does provide significant improvement of the read out time. In addition to switching to the LucR read out, changes made to the overall protocol resulted in the miniaturization of the assay from a 48 to a 96-well plate format, which preserved sample and allowed for the introduction of replicates. The overall assay time was reduced from 13 to 8 days. The assay has a high degree of specificity, and the previously observed non-specific background inhibition in cells from HIV-1 negative volunteers has been reduced dramatically. Importantly, we observed an increase in positive responses, indicating an improvement in sensitivity compared to the original VIA. Currently, only a limited number of “whole-genome” IMC-LucR viruses are available and our efforts will focus on expanding the panel to better evaluate anti-viral breadth. Overall, we believe the IMC LucR VIA provides a platform to assess functional CD8 T-cell responses in large-scale clinical trial testing, which will enhance the ability

Engineered Luciferase Reporter from a Deep Sea Shrimp Utilizing a Novel Imidazopyrazinone Substrate

PubMed Central

2012-01-01

Bioluminescence methodologies have been extraordinarily useful due to their high sensitivity, broad dynamic range, and operational simplicity. These capabilities have been realized largely through incremental adaptations of native enzymes and substrates, originating from luminous organisms of diverse evolutionary lineages. We engineered both an enzyme and substrate in combination to create a novel bioluminescence system capable of more efficient light emission with superior biochemical and physical characteristics. Using a small luciferase subunit (19 kDa) from the deep sea shrimp Oplophorus gracilirostris, we have improved luminescence expression in mammalian cells ∼2.5 million-fold by merging optimization of protein structure with development of a novel imidazopyrazinone substrate (furimazine). The new luciferase, NanoLuc, produces glow-type luminescence (signal half-life >2 h) with a specific activity ∼150-fold greater than that of either firefly (Photinus pyralis) or Renilla luciferases similarly configured for glow-type assays. In mammalian cells, NanoLuc shows no evidence of post-translational modifications or subcellular partitioning. The enzyme exhibits high physical stability, retaining activity with incubation up to 55 °C or in culture medium for >15 h at 37 °C. As a genetic reporter, NanoLuc may be configured for high sensitivity or for response dynamics by appending a degradation sequence to reduce intracellular accumulation. Appending a signal sequence allows NanoLuc to be exported to the culture medium, where reporter expression can be measured without cell lysis. Fusion onto other proteins allows luminescent assays of their metabolism or localization within cells. Reporter quantitation is achievable even at very low expression levels to facilitate more reliable coupling with endogenous cellular processes. PMID:22894855

Dynamic monitoring of p53 translocation to mitochondria for the analysis of specific inhibitors using luciferase-fragment complementation.

PubMed

Noda, Natsumi; Awais, Raheela; Sutton, Robert; Awais, Muhammad; Ozawa, Takeaki

2017-12-01

Intracellular protein translocation plays a pivotal role in regulating complex biological processes, including cell death. The tumor suppressor p53 is a transcription factor activated by DNA damage and oxidative stress that also translocates from the cytosol into the mitochondrial matrix to facilitate necrotic cell death. However, specific inhibitors of p53 mitochondrial translocation are largely unknown. To explore the inhibitors of p53, we developed a bioluminescent probe to monitor p53 translocation from cytosol to mitochondria using luciferase fragment complementation assays. The probe is composed of a novel pair of luciferase fragments, the N-terminus of green click beetle luciferase CBG68 (CBGN) and multiple-complement luciferase fragment (McLuc1). The combination of luciferase fragments showed significant luminescence intensity and high signal-to-background ratio. When the p53 connected with McLuc1 translocates from cytosol into mitochondrial matrix, CBGN in mitochondrial matrix enables to complement with McLuc1, resulting in the restoration of the luminescence. The luminescence intensity was significantly increased under hydrogen peroxide-induced oxidative stress following the complementation of CBGN and McLuc1. Pifithrin-μ, a selective inhibitor of p53 mitochondrial translocation, prevented the mitochondrial translocation of the p53 probe in a concentration-dependent manner. Furthermore, the high luminescence intensity made it easier to visualize the p53 translocation at a single cell level under a bioluminescence microscope. This p53 mitochondrial translocation assay is a new tool for high-throughput screening to identify novel p53 inhibitors, which could be developed as drugs to treat diseases in which necrotic cell death is a major contributor. © 2017 Wiley Periodicals, Inc.

Graphene nanoribbons as a drug delivery agent for lucanthone mediated therapy of glioblastoma multiforme

DOE Office of Scientific and Technical Information (OSTI.GOV)

Chowdhury, Sayan Mullick; Surhland, Cassandra; Sanchez, Zina

We report use of PEG-DSPE coated oxidized graphene nanoribbons (O-GNR-PEG-DSPE) as agent for delivery of anti-tumor drug Lucanthone (Luc) into Glioblastoma Multiformae (GBM) cells targeting base excision repair enzyme APE-1 (Apurinic endonuclease-1). Lucanthone, an endonuclease inhibitor of APE-1, was loaded onto O-GNR-PEG-DSPEs using a simple non-covalent method. We found its uptake by GBM cell line U251 exceeding 67% and 60% in APE-1-overexpressing U251, post 24 hours (h). However, their uptake was ~38% and 29% by MCF-7 and rat glial progenitor cells (CG-4), respectively. TEM analysis of U251 showed large aggregates of O-GNR-PEG-DSPE in vesicles. Luc-O-GNR-PEG-DSPE was significantly toxic to U251more » but showed little / no toxicity when exposed to MCF-7/CG-4 cells. This differential uptake effect can be exploited to use O-GNR-PEG-DSPEs as a vehicle for Luc delivery to GBM, while reducing nonspecific cytotoxicity to the surrounding healthy tissue. In conclusion, cell death in U251 was necrotic, probably due to oxidative degradation of APE-1.« less

Interaction of Osmotic Stress, Temperature, and Abscisic Acid in the Regulation of Gene Expression in Arabidopsis

PubMed Central

Xiong, Liming; Ishitani, Manabu; Zhu, Jian-Kang

1999-01-01

The impact of simultaneous environmental stresses on plants and how they respond to combined stresses compared with single stresses is largely unclear. By using a transgene (RD29A-LUC) consisting of the firefly luciferase coding sequence (LUC) driven by the stress-responsive RD29A promoter, we investigated the interactive effects of temperature, osmotic stress, and the phytohormone abscisic acid (ABA) in the regulation of gene expression in Arabidopsis seedlings. Results indicated that both positive and negative interactions exist among the studied stress factors in regulating gene expression. At a normal growth temperature (22°C), osmotic stress and ABA act synergistically to induce the transgene expression. Low temperature inhibits the response to osmotic stress or to combined treatment of osmotic stress and ABA, whereas low temperature and ABA treatments are additive in inducing transgene expression. Although high temperature alone does not activate the transgene, it significantly amplifies the effects of ABA and osmotic stress. The effect of multiple stresses in the regulation of RD29A-LUC expression in signal transduction mutants was also studied. The results are discussed in the context of cold and osmotic stress signal transduction pathways. PMID:9880362

G protein-coupled receptor 30 down-regulates cofactor expression and interferes with the transcriptional activity of glucocorticoid.

PubMed

Ylikomi, Timo; Vienonen, Annika; Ahola, Tytti M

2004-11-01

G protein-coupled receptor 30 (GPR30) has previously been described to be important in steroid-mediated growth and to inhibit cell proliferation. Here we investigated whether the effect of GPR30 on cell growth is dependent on steroid hormone receptors. We stably introduced GPR30 in immortalized normal mammary epithelial (HME) cells using retroviruses for gene delivery. GPR30 inhibited the growth and proliferation of the cells. They expressed glucocorticoid receptor, but not estrogen or progesterone receptor. GPR30 down-regulated the expression of cofactor transcription intermediary factor 2 (TIF2) analyzed using quantitative RT-PCR analysis, and also diminished the expression of TIF2 at protein level analyzed by Western blotting using nuclear extracts from mammary epithelial cells. When HME cells were transiently transfected with the glucocorticoid response element MMTV-luc reporter plasmid, stable expression of GPR30 resulted in the abolition of ligand-induced transactivation of the promoter. In COS cells, transient transfection of GPR30 with glucocorticoid receptor alpha resulted in an abrogation of the MMTV-luc and GRE-luc reporter activities induced by dexamethasone. The results suggest a novel mechanism by which membrane-initiated signaling interferes with steroid signaling.

Graphene nanoribbons as a drug delivery agent for lucanthone mediated therapy of glioblastoma multiforme

DOE PAGES

Chowdhury, Sayan Mullick; Surhland, Cassandra; Sanchez, Zina; ...

2014-08-13

We report use of PEG-DSPE coated oxidized graphene nanoribbons (O-GNR-PEG-DSPE) as agent for delivery of anti-tumor drug Lucanthone (Luc) into Glioblastoma Multiformae (GBM) cells targeting base excision repair enzyme APE-1 (Apurinic endonuclease-1). Lucanthone, an endonuclease inhibitor of APE-1, was loaded onto O-GNR-PEG-DSPEs using a simple non-covalent method. We found its uptake by GBM cell line U251 exceeding 67% and 60% in APE-1-overexpressing U251, post 24 hours (h). However, their uptake was ~38% and 29% by MCF-7 and rat glial progenitor cells (CG-4), respectively. TEM analysis of U251 showed large aggregates of O-GNR-PEG-DSPE in vesicles. Luc-O-GNR-PEG-DSPE was significantly toxic to U251more » but showed little / no toxicity when exposed to MCF-7/CG-4 cells. This differential uptake effect can be exploited to use O-GNR-PEG-DSPEs as a vehicle for Luc delivery to GBM, while reducing nonspecific cytotoxicity to the surrounding healthy tissue. In conclusion, cell death in U251 was necrotic, probably due to oxidative degradation of APE-1.« less

An Arabidopsis mutant showing reduced feedback inhibition of photosynthesis.

PubMed

Van Oosten, J J; Gerbaud, A; Huijser, C; Dijkwel, P P; Chua, N H; Smeekens, S C

1997-11-01

Many plant genes are responsive to sugars but the mechanisms used by plants to sense sugars are unknown. A genetic approach has been used in Arabidopsis to identify genes involved in perception and transduction of sugar signals. For this purpose, an in vivo reporter system was established consisting of the light- and sugar-regulated plastocyanin promoter, fused to the luciferase coding sequence (PC-LUC construct). At the seedling stage, expression of the PC-LUC gene is repressed by sucrose, and a number of sucrose-uncoupled (sun) mutants were selected in which sucrose is unable to repress the activity of the PC promoter. Three mutants have been characterized in more detail. The sugar analog 2-deoxy-D-glucose (2DG) was used to repress whole plant photosynthesis, PC-LUC gene expression and total ribulose-1,5-bisphosphate activity. It was found that the sun6 mutation makes plants unresponsive to these 2DG-induced effects. Moreover, unlike wild-type plants, sun6 mutants are insensitive to elevated levels of glucose in the growth medium. These findings suggest that the SUN6 gene is active in a hexose-activated signal transduction pathway.

Extratropical Respones to Amazon Deforestation

NASA Astrophysics Data System (ADS)

Badger, A.; Dirmeyer, P.

2014-12-01

Land-use change (LUC) is known to impact local climate conditions through modifications of land-atmosphere interactions. Large-scale LUC, such as Amazon deforestation, could have a significant effect on the local and regional climates. The question remains as to what the global impact of large-scale LUC could be, as previous modeling studies have shown non-local responses due to Amazon deforestation. A common shortcoming in many previous modeling studies is the use of prescribed ocean conditions, which can act as a boundary condition to dampen the global response with respect to changes in the mean and variability. Using fully coupled modeling simulations with the Community Earth System Model version 1.2.0, the Amazon rainforest has been replaced with a distribution of representative tropical crops. Through the modifications of local land-atmosphere interactions, a significant change in the region, both at the surface and throughout the atmosphere, can be quantified. Accompanying these local changes are significant changes to the atmospheric circulation across all scales, thus modifying regional climates in other locales. Notable impacts include significant changes in precipitation, surface fluxes, basin-wide sea surface temperatures and ENSO behavior.

Oil palm for biodiesel in Brazil—risks and opportunities

NASA Astrophysics Data System (ADS)

Englund, Oskar; Berndes, Göran; Persson, U. Martin; Sparovek, Gerd

2015-04-01

Although mainly used for other purposes, and historically mainly established at the expense of tropical forests, oil palm can be the most land efficient feedstock for biodiesel. Large parts of Brazil are suitable for oil palm cultivation and a series of policy initiatives have recently been launched to promote oil palm production. These initiatives are however highly debated both in the parliament and in academia. Here we present results of a high resolution modelling study of opportunities and risks associated with oil palm production for biodiesel in Brazil, under different energy, policy, and infrastructure scenarios. Oil palm was found to be profitable on extensive areas, including areas under native vegetation where establishment would cause large land use change (LUC) emissions. However, some 40-60 Mha could support profitable biodiesel production corresponding to approximately 10% of the global diesel demand, without causing direct LUC emissions or impinging on protected areas. Pricing of LUC emissions could make oil palm production unprofitable on most lands where conversion would impact on native ecosystems and carbon stocks, if the carbon price is at the level 125/tC, or higher.

Insulin-like growth factor-II regulates bone sialoprotein gene transcription.

PubMed

Choe, Jin; Sasaki, Yoko; Zhou, Liming; Takai, Hideki; Nakayama, Yohei; Ogata, Yorimasa

2016-09-01

Insulin-like growth factor-I and -II (IGF-I and IGF-II) have been found in bone extracts of several different species, and IGF-II is the most abundant growth factor stored in bone. Bone sialoprotein (BSP) is a noncollagenous extracellular matrix glycoprotein associated with mineralized connective tissues. In this study, we have investigated the regulation of BSP transcription by IGF-II in rat osteoblast-like ROS17/2.8 cells. IGF-II (50 ng/ml) increased BSP mRNA and protein levels after 6-h stimulation, and enhanced luciferase activities of the constructs pLUC3 (-116 to +60), pLUC4 (-425 to +60), pLUC5 (-801 to +60) and pLUC6 (-938 to +60). Effects of IGF-II were inhibited by tyrosine kinase, extracellular signal-regulated kinase1/2 and phosphatidylinositol 3-kinase inhibitors, and abrogated by 2-bp mutations in cAMP response element (CRE), FGF2 response element (FRE) and homeodomain protein-binding site (HOX). The results of gel shift assays showed that nuclear proteins binding to CRE, FRE and HOX sites were increased by IGF-II (50 ng/ml) at 3 and 6 h. CREB1, phospho-CREB1, c-Fos and c-Jun antibodies disrupted the formation of the CRE-protein complexes. Dlx5 and Runx2 antibodies disrupted the FRE- and HOX-protein complex formations. These studies therefore demonstrated that IGF-II increased BSP transcription by targeting CRE, FRE and HOX elements in the proximal promoter of the rat BSP gene. Moreover, phospho-CREB1, c-Fos, c-Jun, Dlx5 and Runx2 transcription factors appear to be key regulators of IGF-II effects on BSP transcription.

Gravity affects the responsiveness of Runx2 to 1, 25-dihydroxyvitamin D3 (VD3)

NASA Astrophysics Data System (ADS)

Guo, Feima; Dai, Zhongquan; Wu, Feng; Liu, Zhaoxia; Tan, Yingjun; Wan, Yumin; Shang, Peng; Li, Yinghui

2013-03-01

Bone loss resulting from spaceflight is mainly caused by decreased bone formation, and decreased osteoblast proliferation and differentiation. Transcription factor Runx2 plays an important role in osteoblast differentiation and function by responding to microenvironment changes including cytokine and mechanical factors. The effects of 1, 25-dihydroxyvitamin D3 (VD3) on Runx2 in terms of mechanical competence is far less clear. This study describes how gravity affects the response of Runx2 to VD3. A MC3T3-6OSE2-Luc osteoblast model was constructed in which the activity of Runx2 was reflected by reporter luciferase activity identifed by bone-related cytokines. The results showed that luciferase activity in MC3T3-6OSE2-Luc cells transfected with Runx2 was twice that of the vacant vector. Alkaline phosphatase (ALP) activity was increased in MC3T3-6OSE2-Luc cells by different concentrations of IGF-I and BMP2. MC3T3-6OSE2-Luc cells were cultured under simulated microgravity or centrifuge with or without VD3. In simulated microgravity, luciferase activity was decreased after 48 h of clinorotation culture, but increased in the centrifuge culture. Luciferase activity was increased after VD3 treatment in normal conditions and simulated microgravity, the increase in luciferase activity in simulated microgravity was lower than that in the 1 g condition when simultaneously treated with VD3 and higher than that in the centrifuge condition. Co-immunoprecipitation showed that the interaction between the VD3 receptor (VDR) and Runx2 was decreased by simulated microgravity, but increased by centrifugation. From these results, we conclude that gravity affects the response of Runx2 to VD3 which results from an alteration in the interaction between VDR and Runx2 under different gravity conditions.

Developing a new formulation of sodium phenylbutyrate.

PubMed

Guffon, Nathalie; Kibleur, Yves; Copalu, William; Tissen, C; Breitkreutz, Joerg

2012-12-01

Sodium phenylbutyrate (NaPB) is used as a treatment for urea cycle disorders (UCD). However, the available, licensed granule form has an extremely bad taste, which can compromise compliance and metabolic control. A new, taste-masked, coated-granule formulation (Luc 01) under development was characterised for its in vitro taste characteristics, dissolution profiles and bioequivalence compared with the commercial product. Taste, safety and tolerability were also compared in healthy adult volunteers. The in vitro taste profile of NaPB indicated a highly salty and bitter tasting molecule, but Luc 01 released NaPB only after a lag time of ∼10 s followed by a slow release over a few minutes. In contrast, the licensed granules released NaPB immediately. The pharmacokinetic study demonstrated the bioequivalence of a single 5 g dose of the two products in 13 healthy adult volunteers. No statistical difference was seen either for maximal plasma concentration (C(max)) or for area under the plasma concentration-time curve (AUC). CI for C(max) and AUC(0-inf) of NaPB were included in the bioequivalence range of 0.80-1.25. One withdrawal for vomiting and five reports of loss of taste perception (ageusia) were related to the licensed product. Acceptability, bitterness and saltiness assessed immediately after administration indicated a significant preference for Luc 01 (p<0.01), confirming the results of the taste prediction derived from in vitro measurements. In vitro dissolution, in vitro and in vivo taste profiles support the view that the newly developed granules can be swallowed before release of the bitter active substance, thus avoiding stimulation of taste receptors. Moreover, Luc 01 was shown to be bioequivalent to the licensed product. The availability of a taste-masked form should improve compliance which is critical to the efficacy of NaPB treatment in patients with UCD.

The Androgen Metabolite, 5α-Androstane-3β,17β-Diol (3β-Diol), Activates the Oxytocin Promoter Through an Estrogen Receptor-β Pathway

PubMed Central

Hiroi, Ryoko; Lacagnina, Anthony F.; Hinds, Laura R.; Carbone, David G.; Uht, Rosalie M.

2013-01-01

Testosterone has been shown to suppress the acute stress-induced activation of the hypothalamic-pituitary-adrenal axis; however, the mechanisms underlying this response remain unclear. The hypothalamic-pituitary-adrenal axis is regulated by a neuroendocrine subpopulation of medial parvocellular neurons in the paraventricular nucleus of the hypothalamus (PVN). These neurons are devoid of androgen receptors (ARs). Therefore, a possibility is that the PVN target neurons respond to a metabolite in the testosterone catabolic pathway via an AR-independent mechanism. The dihydrotestosterone metabolite, 5α-androstane-3β,17β-diol (3β-diol), binds and activates estrogen receptor-β (ER-β), the predominant ER in the PVN. In the PVN, ER-β is coexpressed with oxytocin (OT). Therefore, we tested the hypothesis that 3β-diol regulates OT expression through ER-β activation. Treatment of ovariectomized rats with estradiol benzoate or 3β-diol for 4 days increased OT mRNA selectively in the midcaudal, but not rostral PVN compared with vehicle-treated controls. 3β-Diol treatment also increased OT mRNA in the hypothalamic N38 cell line in vitro. The functional interactions between 3β-diol and ER-β with the human OT promoter were examined using an OT promoter-luciferase reporter construct (OT-luc). In a dose-dependent manner, 3β-diol treatment increased OT-luc activity when cells were cotransfected with ER-β, but not ER-α. The 3β-diol–induced OT-luc activity was reduced by deletion of the promoter region containing the composite hormone response element (cHRE). Point mutations of the cHRE also prevented OT-luc activation by 3β-diol. These results indicate that 3β-diol induces OT promoter activity via ER-β–cHRE interactions. PMID:23515287

Development of an animal model of progressive vaccinia in nu/nu mice and the use of bioluminescence imaging for assessment of the efficacy of monoclonal antibodies against vaccinial B5 and L1 proteins.

PubMed

Zaitseva, Marina; Thomas, Antonia; Meseda, Clement A; Cheung, Charles Y K; Diaz, Claudia G; Xiang, Yan; Crotty, Shane; Golding, Hana

2017-08-01

Bioluminescence imaging (BLI) was used to follow dissemination of recombinant vaccinia virus (VACV) expressing luciferase (IHD-J-Luc) in BALB/c nu/nu mice treated post-challenge with monoclonal antibodies (MAbs) against L1 and B5 VACV proteins in a model of Progressive Vaccinia (PV). Areas Under the flux Curve (AUC) were calculated for viral loads in multiple organs in individual mice. Following scarification with 10 5  pfu, IHD-J-Luc VACV undergoes fast replication at the injection site and disseminates rapidly to the inguinal lymph nodes followed by spleen, liver, and axillary lymph nodes within 2-3 days and before primary lesions are visible at the site of scarification. Extension of survival in nude mice treated with a combination of anti-B5 and anti-L1 MAbs 24 h post challenge correlated with a significant reduction in viral load at the site of scarification and delayed systemic dissemination. Nude mice reconstituted with 10 4  T cells prior to challenge with IHD-J-Luc, and treated with MAbs post-challenge, survived infection, cleared the virus from all organs and scarification site, and developed anti-VACV IgG and VACV-specific polyfunctional CD8 + T cells that co-expressed the degranulation marker CD107a, and IFNγ and TNFα cytokines. All T cell reconstituted mice survived intranasal re-challenge with IHD-J-Luc (10 4  pfu) two months after the primary infection. Thus, using BLI to monitor VACV replication in a PV model, we showed that anti-VACV MAbs administered post challenge extended survival of nude mice and protected T cell reconstituted nude mice from lethality by reducing replication at the site of scarification and systemic dissemination of VACV. Published by Elsevier B.V.

Proline Coordination with Fatty Acid Synthesis and Redox Metabolism of Chloroplast and Mitochondria.

PubMed

Shinde, Suhas; Villamor, Joji Grace; Lin, Wendar; Sharma, Sandeep; Verslues, Paul E

2016-10-01

Proline (Pro) accumulation is one of the most prominent changes in plant metabolism during drought and low water potential; however, the regulation and function of Pro metabolism remain unclear. We used a combination of forward genetic screening based on a Proline Dehydrogenase1 (PDH1) promoter-luciferase reporter (PDH1 pro :LUC2) and RNA sequencing of the Pro synthesis mutant p5cs1-4 to identify multiple loci affecting Pro accumulation in Arabidopsis (Arabidopsis thaliana). Two mutants having high PDH1 pro :LUC2 expression and increased Pro accumulation at low water potential were found to be alleles of Cytochrome P450, Family 86, Subfamily A, Polypeptide2 (CYP86A2) and Long Chain Acyl Synthetase2 (LACS2), which catalyze two successive steps in very-long-chain fatty acid (VLCFA) synthesis. Reverse genetic experiments found additional VLCFA and lipid metabolism-related mutants with increased Pro accumulation. Altered cellular redox status is a key factor in the coordination of Pro and VLCFA metabolism. The NADPH oxidase inhibitor diphenyleneiodonium (DPI) induced high levels of Pro accumulation and strongly repressed PDH1 pro :LUC2 expression. cyp86a2 and lacs2 mutants were hypersensitive to diphenyleneiodonium but could be reverted to wild-type Pro and PDH1 pro :LUC2 expression by reactive oxygen species scavengers. The coordination of Pro and redox metabolism also was indicated by the altered expression of chloroplast and mitochondria electron transport genes in p5cs1-4 These results show that Pro metabolism is both influenced by and influences cellular redox status via previously unknown coordination with several metabolic pathways. In particular, Pro and VLCFA synthesis share dual roles to help buffer cellular redox status while producing products useful for stress resistance, namely the compatible solute Pro and cuticle lipids. © 2016 American Society of Plant Biologists. All Rights Reserved.

Proline Coordination with Fatty Acid Synthesis and Redox Metabolism of Chloroplast and Mitochondria1[OPEN

PubMed Central

Shinde, Suhas; Villamor, Joji Grace; Lin, Wendar; Verslues, Paul E.

2016-01-01

Proline (Pro) accumulation is one of the most prominent changes in plant metabolism during drought and low water potential; however, the regulation and function of Pro metabolism remain unclear. We used a combination of forward genetic screening based on a Proline Dehydrogenase1 (PDH1) promoter-luciferase reporter (PDH1pro:LUC2) and RNA sequencing of the Pro synthesis mutant p5cs1-4 to identify multiple loci affecting Pro accumulation in Arabidopsis (Arabidopsis thaliana). Two mutants having high PDH1pro:LUC2 expression and increased Pro accumulation at low water potential were found to be alleles of Cytochrome P450, Family 86, Subfamily A, Polypeptide2 (CYP86A2) and Long Chain Acyl Synthetase2 (LACS2), which catalyze two successive steps in very-long-chain fatty acid (VLCFA) synthesis. Reverse genetic experiments found additional VLCFA and lipid metabolism-related mutants with increased Pro accumulation. Altered cellular redox status is a key factor in the coordination of Pro and VLCFA metabolism. The NADPH oxidase inhibitor diphenyleneiodonium (DPI) induced high levels of Pro accumulation and strongly repressed PDH1pro:LUC2 expression. cyp86a2 and lacs2 mutants were hypersensitive to diphenyleneiodonium but could be reverted to wild-type Pro and PDH1pro:LUC2 expression by reactive oxygen species scavengers. The coordination of Pro and redox metabolism also was indicated by the altered expression of chloroplast and mitochondria electron transport genes in p5cs1-4. These results show that Pro metabolism is both influenced by and influences cellular redox status via previously unknown coordination with several metabolic pathways. In particular, Pro and VLCFA synthesis share dual roles to help buffer cellular redox status while producing products useful for stress resistance, namely the compatible solute Pro and cuticle lipids. PMID:27512016

Carbon dynamics on agricultural land reverting to woody land in Ontario, Canada.

PubMed

Voicu, Mihai F; Shaw, Cindy; Kurz, Werner A; Huffman, Ted; Liu, Jiangui; Fellows, Max

2017-05-15

The 2015 Paris Agreement reinforces the importance of the land sector and its contribution to greenhouse gas (GHG) reductions. Thus, there is growing interest in improving estimates of the GHG balance in response to land-use changes (LUCs) involving agriculture and forestry, for national-scale reporting, and for carbon (C) offsets. Large agricultural areas in Europe, Russia and North America are reverting to forest, either naturally or through planting, after abandonment of agricultural land, and this trend may have a substantial impact on carbon budgets. We report results of a pilot project in the Mixedwood Plains ecozone of eastern Canada to analyze the change in the C budget on a landscape over 15 years on abandoned cropland where woody vegetation is regenerating. Thirty-six plots (2 km × 2 km) with paired aerial photographs taken circa 1994 and circa 2008 at a scale of 1:10,000 or larger were randomly selected from the 20 km × 20 km National Forest Inventory (NFI) grid. A spatially-explicit version of the Carbon Budget Model of the Canadian Forest Sector (CBM-CFS3) was used to estimate impacts of LUC on C stocks and fluxes. Polygons identifying areas of LUC within each photo plot were delineated, classified, and evaluated to provide input data for the model. The rate of C accumulation in our study area was found to be relatively constant over the entire simulation period, at 1.07 Mg C/ha/yr. Abandoned agricultural land reverting to woody lands could play an important role in regional and national C sequestration in Canada, but more research is required to quantify the areal extent of this LUC. Crown Copyright © 2017. Published by Elsevier Ltd. All rights reserved.

Norisoboldine, an alkaloid from Radix linderae, inhibits NFAT activation and attenuates 2,4-dinitrofluorobenzene-induced dermatitis in mice.

PubMed

Gao, Shuang; Li, Wencai; Lin, Guochao; Liu, Guangrong; Deng, Wenjuan; Zhai, Chuntao; Bian, Chunliang; He, Gaiying; Hu, Zhenlin

2016-10-01

The nuclear factor of activated T-cells (NFAT) is a family of transcription factors, essential for T-cell activation. Norisoboldine (NOR), an isoquinoline alkaloid from Radix linderae, has been demonstrated to possess anti-inflammatory activity. This study examines NOR's effect on NFAT activation and its therapeutic potential for atopic dermatitis (AD). The transcriptional activity of NFAT was examined with luciferase reporter assay, using K562-luc cells, stimulated with 20 ng/mL PMA plus 1 μM ionomycin. NFAT dephosphorylation was examined by immuno-blotting in K562-luc cells and Jurkat cells. Interleukin-2 (IL-2) expression in Jurkat cells was examined by real-time PCR. A mouse model of dermatitis, induced by 2,4-dinitrochlorobenzene (DNCB), was used to test NOR's therapeutic potential for AD. NOR, dose-dependently, inhibited PMA and ionomycin-induced NFAT reporter gene expression in K562-luc cells in the range of 2-50 μM. NOR also inhibited PMA and ionomycin-induced NFAT dephosphorylation in K562-luc cells and Jurkat cells. Consequently, NOR suppressed PMA plus ionomycin-induced IL-2 expression in Jurkat cells. The administration of NOR (10 mg/kg, i.p.), alleviated DNCB-induced dermatitis in mice, by the reduction of ear swelling and attenuation of inflammatory infiltration into ear tissue. Moreover, mRNA levels of INF-γ, TNF-α, IL-4 and IL-6 in ears of NOR-treated mice were reduced by 78.4, 77.8, 72.3 and 73.9%, respectively, compared with untreated controls. This study demonstrates that NOR inhibits NFAT activation in T-cells and alleviates AD-like inflammatory reaction in a DNCB-induced dermatitis model, highlighting NOR as a potential therapeutic agent for AD.

Influence of spatially dependent, modeled soil carbon emission factors on life-cycle greenhouse gas emissions of corn and cellulosic ethanol

DOE Office of Scientific and Technical Information (OSTI.GOV)

Qin, Zhangcai; Dunn, Jennifer B.; Kwon, Hoyoung

Converting land to biofuel feedstock production incurs changes in soil organic carbon (SOC) that can influence biofuel life-cycle greenhouse gas (GHG) emissions. Estimates of these land use change (LUC) and life-cycle GHG emissions affect biofuels’ attractiveness and eligibility under a number of renewable fuel policies in the U.S. and abroad. Modeling was used to refine the spatial resolution and depth-extent of domestic estimates of SOC change for land (cropland, cropland pasture, grasslands, and forests) conversion scenarios to biofuel crops (corn, corn stover, switchgrass, Miscanthus, poplar, and willow). In most regions, conversions from cropland and cropland pasture to biofuel crops ledmore » to neutral or small levels of SOC sequestration, while conversion of grassland and forest generally caused net SOC loss. Results of SOC change were incorporated into the Greenhouse Gases, Regulated Emissions, and Energy use in Transportation (GREET) model to assess their influence on life-cycle GHG emissions for the biofuels considered. Total LUC GHG emissions (g CO2eq MJ-1) were 2.1–9.3 for corn, -0.7 for corn stover, -3.4–12.9 for switchgrass, and -20.1–-6.2 for Miscanthus; these varied with SOC modeling assumptions applied. Extending soil depth from 30 to 100cm affected spatially-explicit SOC change and overall LUC GHG emissions; however the influence on LUC GHG emissions estimates were less significant in corn and corn stover than cellulosic feedstocks. Total life-cycle GHG emissions (g CO2eq MJ-1, 100cm) were estimated to be 59–66 for corn ethanol, 14 for stover ethanol, 18-26 for switchgrass ethanol, and -0.6–-7 for Miscanthus ethanol.« less

In vivo effects of human adipose-derived stem cells reseeding on acellular bovine pericardium in nude mice.

PubMed

Wu, Qingkai; Dai, Miao; Xu, Peirong; Hou, Min; Teng, Yincheng; Feng, Jie

2016-01-01

Tissue-engineered biologic products may be a viable option in the reconstruction of pelvic organ prolapse (POP). This study was based on the hypothesis that human adipose-derived stem cells (hASCs) are viable in acellular bovine pericardium (ABP), when reseeded by two different techniques, and thus, aid in the reconstruction. To investigate the reseeding of hASCs on ABP grafts by using non-invasive bioluminescence imaging (BLI), and to identify the effective hASCs-scaffold combinations that enabled regeneration. Thirty female athymic nude mice were randomly divided into three groups: In the VIVO group, ABPs were implanted in the subcutaneous pockets and enhanced green fluorescent protein luciferase (eGFP·Luc)-hASCs (1 × 10(6) cells/50 µL) were injected on the ABP at the same time. In the VITRO group, the mice were implanted with grafts that ABP were co-cultured with eGFP·Luc-hASCs in vitro. The BLANK group mice were implanted with ABP only. The eGFP·Luc-hASCs reseeded on ABP were analyzed by BLI, histology, and immunohistochemistry. The eGFP·Luc-hASCs reseeded on ABP could be visualized at 12 weeks in vivo. Histology revealed that the VIVO group displayed the highest cell ingrowths, small vessels, and percent of collagen content per unit area. Desmin and α-smooth muscle actin were positive at the same site in the VIVO group cells. However, few smooth muscles were observed in the VITRO and BLANK groups. These results suggest that hASCs reseeded on ABP in vivo during surgery may further enhance the properties of ABP and may promote regeneration at the recipient site, resulting in a promising treatment option for POP. © 2016 by the Society for Experimental Biology and Medicine.

Bibliography of Soviet Development in Magnetohydrodynamics

DTIC Science & Technology

1976-03-22

re i I I i .._ i ,»,- J \\*^ ’ —* Ak. _ „ ■***"" I 1.1*1 .->• , JanyS«.Dec»--©?^ jT I I I I ! ! 6tuavt| / S » O./i-.u beritt-V. A...for a round casting. MG, no. 3, 1975, 119-123. 53. Ginzburg, A. S ., et al. On the possibility of flow measuring by a J conduction method...regimes. IN: Sb 35, no. 1, 974, 230-243. (RZhElektrotekhenerg, 2/75, 2F32) 86. Medin, S . A. End effect in an optimal plane MHD generator

Newman-Penrose constants of the Kerr-Newman metric

DOE Office of Scientific and Technical Information (OSTI.GOV)

Gong Xuefei; Shang Yu; Bai Shan

The Newman-Unti formalism of the Kerr-Newman metric near future null infinity is developed, with which the Newman-Penrose constants for both the gravitational and electromagnetic fields of the Kerr-Newman metric are computed and shown to be zero. The multipole structure near future null infinity in the sense of Janis-Newman of the Kerr-Newman metric is then further studied. It is found that up to the 2{sup 4}-pole, modulo a constant dependent upon the order of the pole, these multipole moments agree with those of Geroch-Hansen multipole moments defined at spatial infinity.

Effect of external and internal factors on the expression of reporter genes driven by the N resistance gene promoter.

PubMed

Kathiria, Palak; Sidler, Corinne; Woycicki, Rafal; Yao, Youli; Kovalchuk, Igor

2013-07-01

The role of resistance (R) genes in plant pathogen interaction has been studied extensively due to its economical impact on agriculture. Interaction between tobacco mosaic virus (TMV) and the N protein from tobacco is one of the most widely used models to understand various aspects of pathogen resistance. The transcription activity governed by N gene promoter is one of the least understood elements of the model. In this study, the N gene promoter was cloned and fused with two different reporter genes, one encoding β-glucuronidase (N::GUS) and another, luciferase (N::LUC). Tobacco plants transformed with the N::GUS or N::LUC reporter constructs were screened for homozygosity and stable expression. Histochemical analysis of N::GUS tobacco plants revealed that the expression is organ specific and developmentally regulated. Whereas two week old plants expressed GUS in midveins only, 6-wk-old plants also expressed GUS in leaf lamella. Roots did not show GUS expression at any time during development. Experiments to address effects of external stress were performed using N::LUC tobacco plants. These experiments showed that N gene promoter expression was suppressed when plants were exposed to high but not low temperatures. Expression was also upregulated in response to TMV, but no changes were observed in plants treated with SA.

Biofuels that cause land-use change may have much larger non-GHG air quality emissions than fossil fuels.

PubMed

Tsao, C-C; Campbell, J E; Mena-Carrasco, M; Spak, S N; Carmichael, G R; Chen, Y

2012-10-02

Although biofuels present an opportunity for renewable energy production, significant land-use change resulting from biofuels may contribute to negative environmental, economic, and social impacts. Here we examined non-GHG air pollution impacts from both indirect and direct land-use change caused by the anticipated expansion of Brazilian biofuels production. We synthesized information on fuel loading, combustion completeness, and emission factors, and developed a spatially explicit approach with uncertainty and sensitivity analyses to estimate air pollution emissions. The land-use change emissions, ranging from 6.7 to 26.4 Tg PM(2.5), were dominated by deforestation burning practices associated with indirect land-use change. We also found Brazilian sugar cane ethanol and soybean biodiesel including direct and indirect land-use change effects have much larger life-cycle emissions than conventional fossil fuels for six regulated air pollutants. The emissions magnitude and uncertainty decrease with longer life-cycle integration periods. Results are conditional to the single LUC scenario employed here. After LUC uncertainty, the largest source of uncertainty in LUC emissions stems from the combustion completeness during deforestation. While current biofuels cropland burning policies in Brazil seek to reduce life-cycle emissions, these policies do not address the large emissions caused by indirect land-use change.

High-throughput, luciferase-based reverse genetics systems for identifying inhibitors of Marburg and Ebola viruses.

PubMed

Uebelhoer, Luke S; Albariño, César G; McMullan, Laura K; Chakrabarti, Ayan K; Vincent, Joel P; Nichol, Stuart T; Towner, Jonathan S

2014-06-01

Marburg virus (MARV) and Ebola virus (EBOV), members of the family Filoviridae, represent a significant challenge to global public health. Currently, no licensed therapies exist to treat filovirus infections, which cause up to 90% mortality in human cases. To facilitate development of antivirals against these viruses, we established two distinct screening platforms based on MARV and EBOV reverse genetics systems that express secreted Gaussia luciferase (gLuc). The first platform is a mini-genome replicon to screen viral replication inhibitors using gLuc quantification in a BSL-2 setting. The second platform is complementary to the first and expresses gLuc as a reporter gene product encoded in recombinant infectious MARV and EBOV, thereby allowing for rapid quantification of viral growth during treatment with antiviral compounds. We characterized these viruses by comparing luciferase activity to virus production, and validated luciferase activity as an authentic real-time measure of viral growth. As proof of concept, we adapt both mini-genome and infectious virus platforms to high-throughput formats, and demonstrate efficacy of several antiviral compounds. We anticipate that both approaches will prove highly useful in the development of anti-filovirus therapies, as well as in basic research on the filovirus life cycle. Published by Elsevier B.V.

Visualization of Oxidative Stress Induced by Experimental Periodontitis in Keap1-Dependent Oxidative Stress Detector-Luciferase Mice.

PubMed

Kataoka, Kota; Ekuni, Daisuke; Tomofuji, Takaaki; Irie, Koichiro; Kunitomo, Muneyoshi; Uchida, Yoko; Fukuhara, Daiki; Morita, Manabu

2016-11-16

The aim of this study was to investigate whether a Keap1-dependent oxidative stress detector-luciferase (OKD-LUC) mouse model would be useful for the visualization of oxidative stress induced by experimental periodontitis. A ligature was placed around the mandibular first molars for seven days to induce periodontitis. Luciferase activity was measured with an intraperitoneal injection of d-luciferin on days 0, 1, and 7. The luciferase activity in the periodontitis group was significantly greater than that in the control group at seven days. The expressions of heme oxygenase-1 (HO-1) and malondialdehyde in periodontal tissue were significantly higher in the periodontitis group than in the control group. Immunofluorescent analysis confirmed that the nuclear translocation of nuclear factor erythroid 2-related factor 2 (Nrf2) occurred more frequently in the periodontitis group than in the control group. This study found that under oxidative stress induced by experimental periodontitis, the Nrf2/antioxidant defense pathway was activated and could be visualized from the luciferase activity in the OKD-LUC model. Thus, the OKD-LUC mouse model may be useful for exploring the mechanism underlying the relationship between the Nrf2/antioxidant defense pathway and periodontitis by enabling the visualization of oxidative stress over time.

Biosynthesis-inspired deracemizative production of d-luciferin by combining luciferase and thioesterase.

PubMed

Maeda, Juri; Kato, Dai-Ichiro; Okuda, Masatoshi; Takeo, Masahiro; Negoro, Seiji; Arima, Kazunari; Ito, Yuji; Niwa, Kazuki

2017-08-01

Due to the strict enantioselectivity of firefly luciferase, only d-luciferin can be used as a substrate for bioluminescence reactions. Unfortunately, luciferin racemizes easily and accumulation of nonluminous l-luciferin has negative influences on the light emitting reaction. Thus, maintaining the enantiopurity of luciferin in the reaction mixture is one of the most important demands in bioluminescence applications using firefly luciferase. In fireflies, however, l-luciferin is the biosynthetic precursor of d-luciferin, which is produced from the L-form undergoing deracemization. This deracemization consists of three successive reactions: l-enantioselective thioesterification by luciferase, in situ epimerization, and hydrolysis by thioesterase. In this work, we introduce a deracemizative luminescence system inspired by the biosynthetic pathway of d-luciferin using a combination of firefly luciferase from Luciola cruciata (LUC-G) and fatty acyl-CoA thioesterase II from Escherichia coli (TESB). The enzymatic reaction property analysis indicated the importance of the concentration balance between LUC-G and TESB for efficient d-luciferin production and light emission. Using this deracemizative luminescence system, a highly sensitive quantitative analysis method for l-cysteine was constructed. This LUC-G-TESB combination system can improve bioanalysis applications using the firefly bioluminescence reaction by efficient deracemization of D-luciferin. Copyright © 2017 Elsevier B.V. All rights reserved.

A novel method for quantifying the greenhouse gas emissions of biofuels based on historical land use change

NASA Astrophysics Data System (ADS)

Liu, X.; Rhodes, J.; Clarens, A. F.

2012-12-01

Land use change (LUC) emissions have been at the center of an ongoing debate about how the carbon footprint of biofuels compare to petroleum-based fuels over their entire life cycle. The debate about LUC has important implications in the US, the EU, and other countries that are working to deploy biofuel policies, informed by life cycle assessment, that promote carbon emission reductions, among other things. LUC calculations often distinguish between direct land use change (DLUC), those that occur onsite, and indirect land use change (ILUC), those that result from market mechanisms leading to emissions that are either spatially or temporally removed from the agricultural activity. These designations are intended to capture the fundamental connection between agricultural production of biofuel feedstock and its physical effects on the land, but both DLUC and ILUC can be difficult to measure and apply broadly. ILUC estimates are especially challenging to quantify because they rely on global economic models to assess how much land would be brought into production in other countries as a consequence of biofuel feedstock cultivation. As a result, ILUC estimates inherently uncertain, are sensitive to complex assumptions, have limited transparency, and have precipitated sufficient controversy to delay development of coherent biofuel policies. To address these shortcomings of conventional LUC methodologies, we have developed a method for estimating land use change emissions that is based on historical emissions from a parcel of land. The method, which we call historical land use change (HLUC) can be readily quantified for any parcel of land in the world using open source datasets of historical emissions. HLUC is easy to use and is directly tied to the physical processes on land used for biofuel production. The emissions from the HLUC calculations are allocated between historical agricultural activity and proposed biofuel feedstock cultivation. This is compatible with

[Self-esteem in adolescents with and without cleft-lip and/or palate].

PubMed

de Andrade, D; Angerami, E L

2001-01-01

Self-image is an important determinant of self-esteem among men and women, which makes us reflect about the process of rehabilitation of people with facial malformations. In order to measure self-esteem, the Janis and Field Scale was used. The sample consisted of 608 adolescents aged 17 to 20 years including males and females. Two hundred and thirty-five had a cleft lip and/or palate and 373 did not. The analysis of the self-esteem levels obtained enabled to conclude that the adolescents with a cleft lip and/or palate behave differently from those without it, that is, they present lower self-esteem scores.

Mechanism of alcohol-enhanced lucigenin chemiluminescence in alkaline solution.

PubMed

Chi, Quan; Chen, Wanying; He, Zhike

2015-11-01

The chemiluminescence (CL) of lucigenin (Luc(2+)) can be enhanced by different alcohols in alkaline solution. The effect of different fatty alcohols on the CL of lucigenin was related to the carbon chain length and the number of hydroxyl groups. Glycerol provides the greatest enhancement. UV/Vis absorption spectra and fluorescence spectra showed that N-methylacridone (NMA) was produced in the CL reaction in the presence of different alcohols. The peak of the CL spectrum was located at 470 nm in all cases, indicating that the luminophore was always the excited-state NMA. The quenching of lucigenin CL by superoxide dismutase (SOD) and the electron spin resonance (ESR) results with the spin trap of 5,5-dimethyl-1-pyrroline N-oxide (DMPO) demonstrated that superoxide anions (O2 (•-)) were generated from dissolved oxygen in the CL reaction and that glycerol and dihydroxyacetone (DHA) can promote O2 (•-) production by the reduction of dissolved oxygen in alkaline solution. It was assumed that the enhancement provided by different alcohols was related to the solvent effect and reducing capacity. Glycerol and DHA can also reduce Luc(2+) into lucigenin cation radicals (Luc(•+) ), which react with O2 (•-) to produce CL, and glycerol can slowly transform into DHA, which is oxidized quickly in alkaline solution. Copyright © 2015 John Wiley & Sons, Ltd.

Visualization and in vivo tracking of the exosomes of murine melanoma B16-BL6 cells in mice after intravenous injection.

PubMed

Takahashi, Yuki; Nishikawa, Makiya; Shinotsuka, Haruka; Matsui, Yuriko; Ohara, Saori; Imai, Takafumi; Takakura, Yoshinobu

2013-05-20

The development of exosomes as delivery vehicles requires understanding how and where exogenously administered exosomes are distributed in vivo. In the present study, we designed a fusion protein consisting of Gaussia luciferase and a truncated lactadherin, gLuc-lactadherin, and constructed a plasmid expressing the fusion protein. B16-BL6 murine melanoma cells were transfected with the plasmid, and exosomes released from the cells were collected by ultracentrifugation. Strong luciferase activity was detected in the fraction containing exosomes, indicating their efficient labeling with gLuc-lactadherin. Then, the labeled B16-BL6 exosomes were intravenously injected into mice, and their tissue distribution was evaluated. Pharmacokinetic analysis of the exosome blood concentration-time profile revealed that B16-BL6 exosomes disappeared very quickly from the blood circulation with a half-life of approximately 2min. Little luciferase activity was detected in the serum at 4h after exosome injection, suggesting rapid clearance of B16-BL6 exosomes in vivo. Moreover, sequential in vivo imaging revealed that the B16-BL6 exosome-derived signals distributed first to the liver and then to the lungs. These results indicate that gLuc-lactadherin labeling is useful for tracing exosomes in vivo and that B16-BL6 exosomes are rapidly cleared from the blood circulation after systemic administration. Copyright © 2013 Elsevier B.V. All rights reserved.

The role of cyclophosphamide in enhancing antitumor efficacy of an adenovirus oncolytic vector in subcutaneous Syrian hamster tumors

PubMed Central

Young, Brittany A.; Spencer, Jacqueline F.; Ying, Baoling; Tollefson, Ann E.; Toth, Karoly; Wold, William S. M.

2013-01-01

We have previously reported that intratumoral injection of VRX-007—an Ad5-based vector overexpressing ADP (Adenovirus Death Protein)—can suppress the growth of subcutaneous HaK (hamster renal cancer) tumors. VRX-007 replication and tumor growth inhibition are enhanced when the hamsters are immunosuppressed by a high dose of cyclophosphamide (CP), an immunosuppressive and chemotherapeutic agent. Here we report that continuous immunosuppression with CP was not required for increased oncolytic activity of VRX-007 because short-term dosing or continuous dosing with the drug yielded similar antitumor results. Prolonged viral replication was found only in animals on continuous CP treatment. We used 007-Luc, a replication-competent, luciferase-expressing vector similar to VRX-007 to investigate the replication of the vector over time. Tumor growth inhibition was similar in hamsters given CP treatment either one week before or one week after 007-Luc injection, which suggests that CP exerts its antitumor efficacy independently of vector therapy. 007-Luc did not spread far from the inoculation site, even in immunosuppressed, CP-treated animals. Our results indicate that the enhanced effectiveness that is produced by the combination of VRX-007 and CP therapies is due to their two independent mechanisms and that they do not have to be given simultaneously for the improved outcome shown. PMID:23928731

The role of cyclophosphamide in enhancing antitumor efficacy of an adenovirus oncolytic vector in subcutaneous Syrian hamster tumors.

PubMed

Young, B A; Spencer, J F; Ying, B; Tollefson, A E; Toth, K; Wold, W S M

2013-09-01

We have previously reported that intratumoral injection of VRX-007--an Ad5 (a species C adenovirus)-based vector overexpressing adenovirus death protein--can suppress the growth of subcutaneous HaK (hamster renal cancer) tumors. VRX-007 replication and tumor growth inhibition are enhanced when the hamsters are immunosuppressed by a high dose of cyclophosphamide (CP), an immunosuppressive and chemotherapeutic agent. Here, we report that continuous immunosuppression with CP was not required for increased oncolytic activity of VRX-007 because short-term dosing or continuous dosing with the drug yielded similar antitumor results. Prolonged viral replication was found only in animals on continuous CP treatment. We used 007-Luc, a replication-competent, luciferase-expressing vector similar to VRX-007, to investigate the replication of the vector over time. Tumor growth inhibition was similar in hamsters given CP treatment either 1 week before or 1 week after 007-Luc injection, which suggests that CP exerts its antitumor efficacy independently of vector therapy. 007-Luc did not spread far from the inoculation site, even in immunosuppressed, CP-treated animals. Our results indicate that the enhanced effectiveness that is produced by the combination of VRX-007 and CP therapies is due to their two independent mechanisms and that they do not have to be given simultaneously for the improved outcome.

Life cycle greenhouse gas emissions of sugar cane renewable jet fuel.

PubMed

Moreira, Marcelo; Gurgel, Angelo C; Seabra, Joaquim E A

2014-12-16

This study evaluated the life cycle GHG emissions of a renewable jet fuel produced from sugar cane in Brazil under a consequential approach. The analysis included the direct and indirect emissions associated with sugar cane production and fuel processing, distribution, and use for a projected 2020 scenario. The CA-GREET model was used as the basic analytical tool, while Land Use Change (LUC) emissions were estimated employing the GTAP-BIO-ADV and AEZ-EF models. Feedstock production and LUC impacts were evaluated as the main sources of emissions, respectively estimated as 14.6 and 12 g CO2eq/MJ of biofuel in the base case. However, the renewable jet fuel would strongly benefit from bagasse and trash-based cogeneration, which would enable a net life cycle emission of 8.5 g CO2eq/MJ of biofuel in the base case, whereas Monte Carlo results indicate 21 ± 11 g CO2eq/MJ. Besides the major influence of the electricity surplus, the sensitivity analysis showed that the cropland-pasture yield elasticity and the choice of the land use factor employed to sugar cane are relevant parameters for the biofuel life cycle performance. Uncertainties about these estimations exist, especially because the study relies on projected performances, and further studies about LUC are also needed to improve the knowledge about their contribution to the renewable jet fuel life cycle.

Circadian Clock-Regulated Expression of Phytochrome and Cryptochrome Genes in Arabidopsis1

PubMed Central

Tóth, Réka; Kevei, Éva; Hall, Anthony; Millar, Andrew J.; Nagy, Ferenc; Kozma-Bognár, László

2001-01-01

Many physiological and biochemical processes in plants exhibit endogenous rhythms with a period of about 24 h. Endogenous oscillators called circadian clocks regulate these rhythms. The circadian clocks are synchronized to the periodic environmental changes (e.g. day/night cycles) by specific stimuli; among these, the most important is the light. Photoreceptors, phytochromes, and cryptochromes are involved in setting the clock by transducing the light signal to the central oscillator. In this work, we analyzed the spatial, temporal, and long-term light-regulated expression patterns of the Arabidopsis phytochrome (PHYA to PHYE) and cryptochrome (CRY1 and CRY2) promoters fused to the luciferase (LUC+) reporter gene. The results revealed new details of the tissue-specific expression and light regulation of the PHYC and CRY1 and 2 promoters. More importantly, the data obtained demonstrate that the activities of the promoter::LUC+ constructs, with the exception of PHYC::LUC+, display circadian oscillations under constant conditions. In addition, it is shown by measuring the mRNA abundance of PHY and CRY genes under constant light conditions that the circadian control is also maintained at the level of mRNA accumulation. These observations indicate that the plant circadian clock controls the expression of these photoreceptors, revealing the formation of a new regulatory loop that could modulate gating and resetting of the circadian clock. PMID:11743105

Development of a dual luciferase activity and fluorescamine protein assay adapted to a 384 micro-well plate format: Reducing variability in human luciferase transactivation cell lines aimed at endocrine active substances.

PubMed

Brennan, Jennifer C; Tillitt, Donald E

2018-03-01

There is a need to adapt cell bioassays to 384-well and 1536-well formats instead of the traditional 96-well format as high-throughput screening (HTS) demands increase. However, the sensitivity and performance of the bioassay must be re-verified in these higher micro-well plates, and verification of cell health must also be HT (high-throughput). We have adapted two commonly used human breast luciferase transactivation cell bioassays, the recently re-named estrogen agonist/antagonist screening VM7Luc4E2 cell bioassay (previously designated BG1Luc4E2) and the androgen/glucocorticoid screening MDA-kb2 cell bioassay, to 384-well formats for HTS of endocrine-active substances (EASs). This cost-saving adaptation includes a fast, accurate, and easy measurement of protein amount in each well via the fluorescamine assay with which to normalize luciferase activity of cell lysates without requiring any transfer of the cell lysates. Here we demonstrate that by accounting for protein amount in the cell lysates, antagonistic agents can easily be distinguished from cytotoxic agents in the MDA-kb2 and VM7Luc4E2 cell bioassays. Additionally, we demonstrate via the fluorescamine assay improved interpretation of luciferase activity in wells along the edge of the plate (the so-called "edge effect"), thereby increasing usable wells to the entire plate, not just interior wells. Published by Elsevier Ltd.

Development of a dual luciferase activity and fluorescamine protein assay adapted to a 384 micro-well plate format: Reducing variability in human luciferase transactivation cell lines aimed at endocrine active substances

USGS Publications Warehouse

Brennan, Jennifer; Tillitt, Donald E.

2018-01-01

There is a need to adapt cell bioassays to 384-well and 1536-well formats instead of the traditional 96-well format as high-throughput screening (HTS) demands increase. However, the sensitivity and performance of the bioassay must be re-verified in these higher micro-well plates, and verification of cell health must also be HT (high-throughput). We have adapted two commonly used human breast luciferase transactivation cell bioassays, the recently re-named estrogen agonist/antagonist screening VM7Luc4E2 cell bioassay (previously designated BG1Luc4E2) and the androgen/glucocorticoid screening MDA-kb2 cell bioassay, to 384-well formats for HTS of endocrine-active substances (EASs). This cost-saving adaptation includes a fast, accurate, and easy measurement of protein amount in each well via the fluorescamine assay with which to normalize luciferase activity of cell lysates without requiring any transfer of the cell lysates. Here we demonstrate that by accounting for protein amount in the cell lysates, antagonistic agents can easily be distinguished from cytotoxic agents in the MDA-kb2 and VM7Luc4E2 cell bioassays. Additionally, we demonstrate via the fluorescamine assay improved interpretation of luciferase activity in wells along the edge of the plate (the so-called “edge effect”), thereby increasing usable wells to the entire plate, not just interior wells.

Functional repression of PtSND2 represses growth and development by disturbing auxin biosynthesis, transport and signaling in transgenic poplar.

PubMed

Wang, Haihai; Tang, Renjie; Wang, Cuiting; Qi, Qi; Gai, Ying; Jiang, Xiangning; Zhang, Hongxia

2015-01-01

Using chimeric repressor silencing technology, we previously reported that functional repression of PtSND2 severely arrested wood formation in transgenic poplar (Populus). Here, we provide further evidence that auxin biosynthesis, transport and signaling were disturbed in these transgenic plants, leading to pleiotropic defects in their growth patterns, including inhibited leaf enlargement and vascular tissue development in the leaf central vein, suppressed cambial growth and fiber elongation in the stem, and arrested growth in the root system. Two transgenic lines, which displayed the most remarkable phenotypic deviation from the wild-type, were selected for detailed studies. In both transgenic lines, expression of genes for auxin biosynthesis, transport and signaling was down-regulated, and indole-3-acetic acid distribution was severely disturbed in the apical buds, leaves, stems and roots of field-grown transgenic plants. Transient transcription dual-luciferase assays of ProPtTYDC2::LUC, ProPttLAX2::LUC and ProPoptrIAA20.2::LUC in poplar protoplasts revealed that expression of auxin-related genes might be regulated by PtSND2 at the transcriptional level. All these results indicate that functional repression of PtSND2 altered auxin biosynthesis, transport and signaling, and thereby disturbed the normal growth and development of transgenic plants. © The Author 2014. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

Different Types of Luciferase Reporters Show Distinct Susceptibility to T3-Evoked Downregulation.

PubMed

Kollár, Anna; Kvárta-Papp, Zsuzsanna; Egri, Péter; Gereben, Balázs

2016-01-01

The firefly luciferase reporter protein is a crucial tool for studies targeting a broad range of biological questions. Importantly, luciferase assays are also widely used to explore mechanisms underlying thyroid hormone dependent regulation of gene expression. However, it was demonstrated that the firefly luciferase reporter is subject to triiodothyronine (T3)-evoked, promoter independent downregulation that is mediated by the thyroid hormone receptor. Since this effect can interfere with readout accuracy, the study aimed to find luciferase reporters that are not susceptible to this phenomenon. Luciferase reporter constructs were generated under the control of a minimal thymidine kinase (TK) promoter and transiently transfected into JEG-3 cells to test their activity upon T3 treatment. Activity of the TK-(dCpG)Luc encoding a synthetic (dCpG)Luciferase and TK-NanoLuc expressing the NanoLuc reporter was not significantly changed by T3 treatment while the firefly luciferase control was suppressed by ∼2.6-fold. T3 also downregulated the activity of Renilla luciferase by ∼30%. Novel types of luciferase reporters, especially the synthetic (dCpG)Luciferase, can be more accurate to study T3-regulated gene expression than the classical firefly luciferase reporter. Renilla luciferase, a popular transfection control of dual luciferase assays, should be used with caution in conditions with T3 treatment.

The impact of historical land use change from 1850 to 2000 on secondary particulate matter and ozone

NASA Astrophysics Data System (ADS)

Heald, Colette L.; Geddes, Jeffrey A.

2016-12-01

Anthropogenic land use change (LUC) since preindustrial (1850) has altered the vegetation distribution and density around the world. We use a global model (GEOS-Chem) to assess the attendant changes in surface air quality and the direct radiative forcing (DRF). We focus our analysis on secondary particulate matter and tropospheric ozone formation. The general trend of expansion of managed ecosystems (croplands and pasturelands) at the expense of natural ecosystems has led to an 11 % decline in global mean biogenic volatile organic compound emissions. Concomitant growth in agricultural activity has more than doubled ammonia emissions and increased emissions of nitrogen oxides from soils by more than 50 %. Conversion to croplands has also led to a widespread increase in ozone dry deposition velocity. Together these changes in biosphere-atmosphere exchange have led to a 14 % global mean increase in biogenic secondary organic aerosol (BSOA) surface concentrations, a doubling of surface aerosol nitrate concentrations, and local changes in surface ozone of up to 8.5 ppb. We assess a global mean LUC-DRF of +0.017, -0.071, and -0.01 W m-2 for BSOA, nitrate, and tropospheric ozone, respectively. We conclude that the DRF and the perturbations in surface air quality associated with LUC (and the associated changes in agricultural emissions) are substantial and should be considered alongside changes in anthropogenic emissions and climate feedbacks in chemistry-climate studies.

Trading forests: land-use change and carbon emissions embodied in production and exports of forest-risk commodities

NASA Astrophysics Data System (ADS)

Henders, Sabine; Persson, U. Martin; Kastner, Thomas

2015-12-01

Production of commercial agricultural commodities for domestic and foreign markets is increasingly driving land clearing in tropical regions, creating links and feedback effects between geographically separated consumption and production locations. Such teleconnections are commonly studied through calculating consumption footprints and quantifying environmental impacts embodied in trade flows, e.g., virtual water and land, biomass, or greenhouse gas emissions. The extent to which land-use change (LUC) and associated carbon emissions are embodied in the production and export of agricultural commodities has been less studied. Here we quantify tropical deforestation area and carbon emissions from LUC induced by the production and the export of four commodities (beef, soybeans, palm oil, and wood products) in seven countries with high deforestation rates (Argentina, Bolivia, Brazil, Paraguay, Indonesia, Malaysia, and Papua New Guinea). We show that in the period 2000-2011, the production of the four analyzed commodities in our seven case countries was responsible for 40% of total tropical deforestation and resulting carbon losses. Over a third of these impacts was embodied in exports in 2011, up from a fifth in 2000. This trend highlights the growing influence of global markets in deforestation dynamics. Main flows of embodied LUC are Latin American beef and soybean exports to markets in Europe, China, the former Soviet bloc, the Middle East and Northern Africa, whereas embodied emission flows are dominated by Southeast Asian exports of palm oil and wood products to consumers in China, India and the rest of Asia, as well as to the European Union. Our findings illustrate the growing role that global consumers play in tropical LUC trajectories and highlight the need for demand-side policies covering whole supply chains. We also discuss the limitations of such demand-side measures and call for a combination of supply- and demand-side policies to effectively limit tropical

Historical emissions critical for mapping decarbonization pathways

NASA Astrophysics Data System (ADS)

Majkut, J.; Kopp, R. E.; Sarmiento, J. L.; Oppenheimer, M.

2016-12-01

Policymakers have set a goal of limiting temperature increase from human influence on the climate. This motivates the identification of decarbonization pathways to stabilize atmospheric concentrations of CO2. In this context, the future behavior of CO2 sources and sinks define the CO2 emissions necessary to meet warming thresholds with specified probabilities. We adopt a simple model of the atmosphere-land-ocean carbon balance to reflect uncertainty in how natural CO2 sinks will respond to increasing atmospheric CO2 and temperature. Bayesian inversion is used to estimate the probability distributions of selected parameters of the carbon model. Prior probability distributions are chosen to reflect the behavior of CMIP5 models. We then update these prior distributions by running historical simulations of the global carbon cycle and inverting with observationally-based inventories and fluxes of anthropogenic carbon in the ocean and atmosphere. The result is a best-estimate of historical CO2 sources and sinks and a model of how CO2 sources and sinks will vary in the future under various emissions scenarios, with uncertainty. By linking the carbon model to a simple climate model, we calculate emissions pathways and carbon budgets consistent with meeting specific temperature thresholds and identify key factors that contribute to remaining uncertainty. In particular, we show how the assumed history of CO2 emissions from land use change (LUC) critically impacts estimates of the strength of the land CO2 sink via CO2 fertilization. Different estimates of historical LUC emissions taken from the literature lead to significantly different parameterizations of the carbon system. High historical CO2 emissions from LUC lead to a more robust CO2 fertilization effect, significantly lower future atmospheric CO2 concentrations, and an increased amount of CO2 that can be emitted to satisfy temperature stabilization targets. Thus, in our model, historical LUC emissions have a

Synthetic versions of firefly luciferase and Renilla luciferase reporter genes that resist transgene silencing in sugarcane

PubMed Central

2014-01-01

Background Down-regulation or silencing of transgene expression can be a major hurdle to both molecular studies and biotechnology applications in many plant species. Sugarcane is particularly effective at silencing introduced transgenes, including reporter genes such as the firefly luciferase gene. Synthesizing transgene coding sequences optimized for usage in the host plant is one method of enhancing transgene expression and stability. Using specified design rules we have synthesised new coding sequences for both the firefly luciferase and Renilla luciferase reporter genes. We have tested these optimized versions for enhanced levels of luciferase activity and for increased steady state luciferase mRNA levels in sugarcane. Results The synthetic firefly luciferase (luc*) and Renilla luciferase (Renluc*) coding sequences have elevated G + C contents in line with sugarcane codon usage, but maintain 75% identity to the native firefly or Renilla luciferase nucleotide sequences and 100% identity to the protein coding sequences. Under the control of the maize pUbi promoter, the synthetic luc* and Renluc* genes yielded 60x and 15x higher luciferase activity respectively, over the native firefly and Renilla luciferase genes in transient assays on sugarcane suspension cell cultures. Using a novel transient assay in sugarcane suspension cells combining co-bombardment and qRT-PCR, we showed that synthetic luc* and Renluc* genes generate increased transcript levels compared to the native firefly and Renilla luciferase genes. In stable transgenic lines, the luc* transgene generated significantly higher levels of expression than the native firefly luciferase transgene. The fold difference in expression was highest in the youngest tissues. Conclusions We developed synthetic versions of both the firefly and Renilla luciferase reporter genes that resist transgene silencing in sugarcane. These transgenes will be particularly useful for evaluating the expression patterns conferred

Synthetic versions of firefly luciferase and Renilla luciferase reporter genes that resist transgene silencing in sugarcane.

PubMed

Chou, Ting-Chun; Moyle, Richard L

2014-04-08

Down-regulation or silencing of transgene expression can be a major hurdle to both molecular studies and biotechnology applications in many plant species. Sugarcane is particularly effective at silencing introduced transgenes, including reporter genes such as the firefly luciferase gene.Synthesizing transgene coding sequences optimized for usage in the host plant is one method of enhancing transgene expression and stability. Using specified design rules we have synthesised new coding sequences for both the firefly luciferase and Renilla luciferase reporter genes. We have tested these optimized versions for enhanced levels of luciferase activity and for increased steady state luciferase mRNA levels in sugarcane. The synthetic firefly luciferase (luc*) and Renilla luciferase (Renluc*) coding sequences have elevated G + C contents in line with sugarcane codon usage, but maintain 75% identity to the native firefly or Renilla luciferase nucleotide sequences and 100% identity to the protein coding sequences.Under the control of the maize pUbi promoter, the synthetic luc* and Renluc* genes yielded 60x and 15x higher luciferase activity respectively, over the native firefly and Renilla luciferase genes in transient assays on sugarcane suspension cell cultures.Using a novel transient assay in sugarcane suspension cells combining co-bombardment and qRT-PCR, we showed that synthetic luc* and Renluc* genes generate increased transcript levels compared to the native firefly and Renilla luciferase genes.In stable transgenic lines, the luc* transgene generated significantly higher levels of expression than the native firefly luciferase transgene. The fold difference in expression was highest in the youngest tissues. We developed synthetic versions of both the firefly and Renilla luciferase reporter genes that resist transgene silencing in sugarcane. These transgenes will be particularly useful for evaluating the expression patterns conferred by existing and newly isolated

Impact of anthropogenic climate change and human activities on environment and ecosystem services in arid regions.

PubMed

Mahmoud, Shereif H; Gan, Thian Y

2018-08-15

The implications of anthropogenic climate change, human activities and land use change (LUC) on the environment and ecosystem services in the coastal regions of Saudi Arabia were analyzed. Earth observations data was used to drive land use categories between 1970 and 2014. Next, a Markov-CA model was developed to characterize the dynamic of LUC between 2014 and 2100 and their impacts on regions' climate and environment. Non-parametric change point and trend detection algorithms were applied to temperature, precipitation and greenhouse gases data to investigate the presence of anthropogenic climate change. Lastly, climate models were used to project future climate change between 2014 and 2100. The analysis of LUC revealed that between 1970 and 2014, built up areas experienced the greatest growth during the study period, leading to a significant monotonic trend. Urban areas increased by 2349.61km 2 between 1970 and 2014, an average increase of >53.4km 2 /yr. The projected LUC between 2014 and 2100 indicate a continued increase in urban areas and irrigated cropland. Human alteration of land use from natural vegetation and forests to other uses after 1970, resulted in a loss, degradation, and fragmentation, all of which usually have devastating effects on the biodiversity of the region. Resulting in a statistically significant change point in temperature anomaly after 1968 with a warming trend of 0.24°C/decade and a downward trend in precipitation anomaly of 12.2mm/decade. Total greenhouse gas emissions including all anthropogenic sources showed a statistically significant positive trend of 78,090Kt/decade after 1991. This is reflected in the future projection of temperature anomaly between 1900 and 2100 with a future warming trend of 0.19°C/decade. In conclusion, human activities, industrial revelation, deforestation, land use transformation and increase in greenhouse gases had significant implications on the environment and ecosystem services of the study area

Transfection and heat-inducible expression of molluscan promoter-luciferase reporter gene constructs in the Biomphalaria glabrata embryonic snail cell line.

PubMed

Yoshino, T P; Wu, X J; Liu, H D

1998-09-01

Studies were initiated to begin developing a genetic transformation system for cells derived from the freshwater gastropod, Biomphalaria glabrata, an intermediate host of the human blood fluke Schistosoma mansoni. Using a 70-kD heat-shock protein (HSP70) cDNA probe obtained from the B. glabrata embryonic (Bge) cell line, we cloned from Bge cells a complete HSP70 gene including a 1-kb genomic DNA fragment in its 5'-flanking region containing sequences indicative of a HSP promoter. Identified in the 5'-half (416 nucleotides) of this genomic fragment were TATA and CAAT boxes, two putative transcription initiation sites, and a series of palindromic DNA repeats with shared homology to the heat-shock element consensus sequence (Bge HSP70(0.5k) promoter). The 3'-half of this upstream flanking region was comprised of a 508-base intron located immediately 5' of the ATG start codon. To determine the functionality of the putative snail promoter sequence, Bge HSP promoter/luciferase (Luc) reporter gene constructs were introduced into Bge cells by N-(1-(2,3-dioleoyloxy) propyl)-N,N,N-trimethylammonium methylsulfate (DOTAP)-mediated transfection methods, and assayed for Luc activity 48 hr following a 1.5-hr heat-shock treatment (40 degrees C). Compared with control vectors or the Bge HSP70(0.5k/1.0k) promoter constructs at 26 degrees C, a 10- to 300-fold increase in Luc expression was obtained only in the Bge HSP70 promoter/Luc-transfected cells following heat-shock. Results of transfection experiments demonstrate that the Bge HSP70(0.5k) DNA segment contains appropriate promoter sequences for driving temperature-inducible gene expression in the Bge snail cell line. This report represents the first isolation and functional characterization of an inducible promoter from a freshwater gastropod mollusc. Successful transient expression of a foreign reporter gene in Bge cells using a homologous, inducible promoter sequence now paves the way for development of methods for stable

Cellular uptake and cytotoxicity of a near-IR fluorescent corrole-TiO2 nanoconjugate.

PubMed

Blumenfeld, Carl M; Sadtler, Bryce F; Fernandez, G Esteban; Dara, Lily; Nguyen, Cathie; Alonso-Valenteen, Felix; Medina-Kauwe, Lali; Moats, Rex A; Lewis, Nathan S; Grubbs, Robert H; Gray, Harry B; Sorasaenee, Karn

2014-11-01

We are investigating the biological and biomedical imaging roles and impacts of fluorescent metallocorrole-TiO2 nanoconjugates as potential near-infrared optical contrast agents in vitro in cancer and normal cell lines. The TiO2 nanoconjugate labeled with the small molecule 2,17-bis(chlorosulfonyl)-5,10,15-tris(pentafluorophenyl)corrolato aluminum(III) (1-Al-TiO2) was prepared. The nanoparticle 1-Al-TiO2 was characterized by transmission electron microscopy (TEM) and integrating-sphere electronic absorption spectroscopy. TEM images of three different samples of TiO2 nanoparticles (bare, H2O2 etched, and 1-Al functionalized) showed similarity in shapes and sizes with an average diameter of 29nm for 1-Al-TiO2. Loading of 1-Al on the TiO2 surfaces was determined to be ca. 20-40mg 1-Al/g TiO2. Confocal fluorescence microscopy (CFM) studies of luciferase-transfected primary human glioblastoma U87-Luc cells treated with the nanoconjugate 1-Al-TiO2 as the contrast agent in various concentrations were performed. The CFM images revealed that 1-Al-TiO2 was found inside the cancer cells even at low doses (0.02-2μg/mL) and localized in the cytosol. Bioluminescence studies of the U87-Luc cells exposed to various amounts of 1-Al-TiO2 showed minimal cytotoxic effects even at higher doses (2-2000μg/mL) after 24h. A similar observation was made using primary mouse hepatocytes (PMH) treated with 1-Al-TiO2 at low doses (0.0003-3μg/mL). Longer incubation times (after 48 and 72h for U87-Luc) and higher doses (>20μg/mL 1-Al-TiO2 for U87-Luc and >3μg/mL 1-Al-TiO2 for PMH) showed decreased cell viability. Copyright © 2014 Elsevier Inc. All rights reserved.

Limited protection of macro-aggregate-occluded organic carbon in Siberian steppe soils

NASA Astrophysics Data System (ADS)

Bischoff, Norbert; Mikutta, Robert; Shibistova, Olga; Puzanov, Alexander; Silanteva, Marina; Grebennikova, Anna; Fuß, Roland; Guggenberger, Georg

2017-05-01

Macro-aggregates especially in agricultural steppe soils are supposed to play a vital role for soil organic carbon (OC) stabilization at a decadal timescale. While most research on soil OC stabilization in steppes focused on North American prairie soils of the Great Plains with information mainly provided by short-term incubation experiments, little is known about the agricultural steppes in southwestern Siberia, though they belong to the greatest conversion areas in the world and occupy an area larger than that in the Great Plains. To quantify the proportion of macro-aggregate-protected OC under different land use as function of land use intensity and time since land use change (LUC) from pasture to arable land in Siberian steppe soils, we determined OC mineralization rates of intact (250-2000 µm) and crushed (< 250 µm) macro-aggregates in long-term incubations over 401 days (20 °C; 60 % water holding capacity) along two agricultural chronosequences in the Siberian Kulunda steppe. Additionally, we incubated bulk soil (< 2000 µm) to determine the effect of LUC and subsequent agricultural use on a fast and a slow soil OC pool (labile vs. more stable OC), as derived from fitting exponential-decay models to incubation data. We hypothesized that (i) macro-aggregate crushing leads to increased OC mineralization due to an increasing microbial accessibility of a previously occluded labile macro-aggregate OC fraction, and (ii) bulk soil OC mineralization rates and the size of the fast OC pool are higher in pasture than in arable soils with decreasing bulk soil OC mineralization rates and size of the fast OC pool as land use intensity and time since LUC increase. Against our hypothesis, OC mineralization rates of crushed macro-aggregates were similar to those of intact macro-aggregates under all land use regimes. Macro-aggregate-protected OC was almost absent and accounted for < 1 % of the total macro-aggregate OC content and to a maximum of 8 ± 4 % of mineralized OC

Heat Shock Protein 70 Enhances Mucosal Immunity against Human Norovirus When Coexpressed from a Vesicular Stomatitis Virus Vector

PubMed Central

Ma, Yuanmei; Duan, Yue; Wei, Yongwei; Liang, Xueya; Niewiesk, Stefan; Oglesbee, Michael

2014-01-01

ABSTRACT Human norovirus (NoV) accounts for 95% of nonbacterial gastroenteritis worldwide. Currently, there is no vaccine available to combat human NoV as it is not cultivable and lacks a small-animal model. Recently, we demonstrated that recombinant vesicular stomatitis virus (rVSV) expressing human NoV capsid protein (rVSV-VP1) induced strong immunities in mice (Y. Ma and J. Li, J. Virol. 85:2942–2952, 2011). To further improve the safety and efficacy of the vaccine candidate, heat shock protein 70 (HSP70) was inserted into the rVSV-VP1 backbone vector. A second construct was generated in which the firefly luciferase (Luc) gene was inserted in place of HSP70 as a control for the double insertion. The resultant recombinant viruses (rVSV-HSP70-VP1 and rVSV-Luc-VP1) were significantly more attenuated in cell culture and viral spread in mice than rVSV-VP1. At the inoculation dose of 1.0 × 106 PFU, rVSV-HSP70-VP1 triggered significantly higher vaginal IgA than rVSV-VP1 and significantly higher fecal and vaginal IgA responses than rVSV-Luc-VP1, although serum IgG and T cell responses were similar. At the inoculation dose of 5.0 × 106 PFU, rVSV-HSP70-VP1 stimulated significantly higher T cell, fecal, and vaginal IgA responses than rVSV-VP1. Fecal and vaginal IgA responses were also significantly increased when combined vaccination of rVSV-VP1 and rVSV-HSP70 was used. Collectively, these data indicate that (i) insertion of an additional gene (HSP70 or Luc) into the rVSV-VP1 backbone further attenuates the VSV-based vaccine in vitro and in vivo, thus improving the safety of the vaccine candidate, and (ii) HSP70 enhances the human NoV-specific mucosal and T cell immunities triggered by a VSV-based human NoV vaccine. IMPORTANCE Human norovirus (NoV) is responsible for more than 95% of acute nonbacterial gastroenteritis worldwide. Currently, there is no vaccine for this virus. Development of a live attenuated vaccine for human NoV has not been possible because it is

Headache

MedlinePlus

... ed. Philadelphia, PA: Elsevier Mosby; 2014:chap 30. Review Date 11/22/2017 Updated by: Luc Jasmin, ... of Maxillofacial Surgery at UCSF, San Francisco, CA. Review provided by VeriMed Healthcare Network. Also reviewed by ...

Cluster headache

MedlinePlus

... ed. Philadelphia, PA: Elsevier Mosby; 2014:chap 30. Review Date 11/22/2017 Updated by: Luc Jasmin, ... of Maxillofacial Surgery at UCSF, San Francisco, CA. Review provided by VeriMed Healthcare Network. Also reviewed by ...

American Idol's Randy Jackson He doesn't miss a beat controlling his diabetes

MedlinePlus

... Jackson is a well-known name in the music world. He has played bass guitar with such musical legends as jazz violinist Jean Luc Ponty, the pop-rock band Journey, and many others. And he's produced ...

Reproductive Biology and Its Impact on Body Size: Comparative Analysis of Mammalian, Avian and Dinosaurian Reproduction

PubMed Central

Werner, Jan; Griebeler, Eva Maria

2011-01-01

Janis and Carrano (1992) suggested that large dinosaurs might have faced a lower risk of extinction under ecological changes than similar-sized mammals because large dinosaurs had a higher potential reproductive output than similar-sized mammals (JC hypothesis). First, we tested the assumption underlying the JC hypothesis. We therefore analysed the potential reproductive output (reflected in clutch/litter size and annual offspring number) of extant terrestrial mammals and birds (as “dinosaur analogs”) and of extinct dinosaurs. With the exception of rodents, the differences in the reproductive output of similar-sized birds and mammals proposed by Janis and Carrano (1992) existed even at the level of single orders. Fossil dinosaur clutches were larger than litters of similar-sized mammals, and dinosaur clutch sizes were comparable to those of similar-sized birds. Because the extinction risk of extant species often correlates with a low reproductive output, the latter difference suggests a lower risk of population extinction in dinosaurs than in mammals. Second, we present a very simple, mathematical model that demonstrates the advantage of a high reproductive output underlying the JC hypothesis. It predicts that a species with a high reproductive output that usually faces very high juvenile mortalities will benefit more strongly in terms of population size from reduced juvenile mortalities (e.g., resulting from a stochastic reduction in population size) than a species with a low reproductive output that usually comprises low juvenile mortalities. Based on our results, we suggest that reproductive strategy could have contributed to the evolution of the exceptional gigantism seen in dinosaurs that does not exist in extant terrestrial mammals. Large dinosaurs, e.g., the sauropods, may have easily sustained populations of very large-bodied species over evolutionary time. PMID:22194835

Reproductive biology and its impact on body size: comparative analysis of mammalian, avian and dinosaurian reproduction.

PubMed

Werner, Jan; Griebeler, Eva Maria

2011-01-01

Janis and Carrano (1992) suggested that large dinosaurs might have faced a lower risk of extinction under ecological changes than similar-sized mammals because large dinosaurs had a higher potential reproductive output than similar-sized mammals (JC hypothesis). First, we tested the assumption underlying the JC hypothesis. We therefore analysed the potential reproductive output (reflected in clutch/litter size and annual offspring number) of extant terrestrial mammals and birds (as "dinosaur analogs") and of extinct dinosaurs. With the exception of rodents, the differences in the reproductive output of similar-sized birds and mammals proposed by Janis and Carrano (1992) existed even at the level of single orders. Fossil dinosaur clutches were larger than litters of similar-sized mammals, and dinosaur clutch sizes were comparable to those of similar-sized birds. Because the extinction risk of extant species often correlates with a low reproductive output, the latter difference suggests a lower risk of population extinction in dinosaurs than in mammals. Second, we present a very simple, mathematical model that demonstrates the advantage of a high reproductive output underlying the JC hypothesis. It predicts that a species with a high reproductive output that usually faces very high juvenile mortalities will benefit more strongly in terms of population size from reduced juvenile mortalities (e.g., resulting from a stochastic reduction in population size) than a species with a low reproductive output that usually comprises low juvenile mortalities. Based on our results, we suggest that reproductive strategy could have contributed to the evolution of the exceptional gigantism seen in dinosaurs that does not exist in extant terrestrial mammals. Large dinosaurs, e.g., the sauropods, may have easily sustained populations of very large-bodied species over evolutionary time.

Persistence, population dynamics and competitiveness for nodulation of marker gene-tagged Rhizobium galegae strains in field lysimeters in the boreal climatic zone.

PubMed

Pitkäjärvi, Jyrki; Räsänen, Leena A; Langenskiöld, Jenny; Wallenius, Kaisa; Niemi, Maarit; Lindström, Kristina

2003-10-01

Abstract A non-indigenous wild-type strain Rhizobium galegae HAMBI 540, which specifically nodulates perennial goat's rue (Galega orientalis), and its marker gene-tagged derivatives R. galegae HAMBI 2363(luc), R. galegae HAMBI 2368(gusA21) and R. galegae HAMBI 2364(gusA30) were used to evaluate the persistence, population dynamics and competitiveness for nodulation of rhizobia under field conditions in Finland. The lysimeters were filled with clean or diesel oil-polluted (3000 mug g(-1)) agricultural soil. During the first 2 years of the field release luc- and gusA21-tagged strains could be effectively detected by cultivation, reinforced with colony polymerase chain reaction. The population densities remained relatively stable from 10(4) to 10(5) cfu g(-1) dry soil from spring until late autumn. Replicate limiting dilution polymerase chain reaction analysis gave comparable results with cultivation with strain HAMBI 2363 until 49 weeks after inoculation. GUS activity of strain HAMBI 2368 could be stably detected in nodules and soil. On the other hand, luc activity weakened clearly in cold conditions along with decreased metabolic activity of rhizobia. The competitive ability for nodulation of the gusA30-tagged strain decreased slowly with time compared to the wild-type strain. Moderate soil pollution did not have significant effects on target bacteria or plant growth. Limited vertical movement of target bacteria outside the rhizosphere was detected from percolated water.

1.5 °C carbon budget dependent on carbon cycle uncertainty and future non-CO2 forcing.

PubMed

Mengis, Nadine; Partanen, Antti-Ilari; Jalbert, Jonathan; Matthews, H Damon

2018-04-11

Estimates of the 1.5 °C carbon budget vary widely among recent studies, emphasizing the need to better understand and quantify key sources of uncertainty. Here we quantify the impact of carbon cycle uncertainty and non-CO 2 forcing on the 1.5 °C carbon budget in the context of a prescribed 1.5 °C temperature stabilization scenario. We use Bayes theorem to weight members of a perturbed parameter ensemble with varying land and ocean carbon uptake, to derive an estimate for the fossil fuel (FF) carbon budget of 469 PgC since 1850, with a 95% likelihood range of (411,528) PgC. CO 2 emissions from land-use change (LUC) add about 230 PgC. Our best estimate of the total (FF + LUC) carbon budget for 1.5 °C is therefore 699 PgC, which corresponds to about 11 years of current emissions. Non-CO 2 greenhouse gas and aerosol emissions represent equivalent cumulative CO 2 emissions of about 510 PgC and -180 PgC for 1.5 °C, respectively. The increased LUC, high non-CO 2 emissions and decreased aerosols in our scenario, cause the long-term FF carbon budget to decrease following temperature stabilization. In this scenario, negative emissions would be required to compensate not only for the increasing non-CO 2 climate forcing, but also for the declining natural carbon sinks.

Medical treatment of orthotopic glioblastoma with transferrin-conjugated nanoparticles encapsulating zoledronic acid.

PubMed

Porru, Manuela; Zappavigna, Silvia; Salzano, Giuseppina; Luce, Amalia; Stoppacciaro, Antonella; Balestrieri, Maria Luisa; Artuso, Simona; Lusa, Sara; De Rosa, Giuseppe; Leonetti, Carlo; Caraglia, Michele

2014-11-15

Glioblastomas are highly aggressive adult brain tumors with poor clinical outcome. In the central nervous system (CNS) the blood-brain barrier (BBB) is the most important limiting factor for both development of new drugs and drug delivery. Here, we propose a new strategy to treat glioblastoma based on transferrin (Tf)-targeted self-assembled nanoparticles (NPs) incorporating zoledronic acid (ZOL) (NPs-ZOL-Tf). NPs-ZOL-Tf have been assessed on the glioblastoma cell line U373MG-LUC that showed a refractoriness in vitro to temozolomide (TMZ) and fotemustine (FTM). NPs-ZOL-Tf treatment resulted in higher in vitro cytotoxic activity than free ZOL. However, the potentiation of anti-proliferative activity of NPs-ZOL-Tf was superimposable to that one induced by NPs-ZOL (not armed with Tf). On the other hand, NPs-ZOL-Tf showed a higher antitumor efficacy if compared with that one caused by NPs-ZOL in immunosuppressed mice intramuscularly bearing U373MG-LUC xenografts, inducing a significant tumor weight inhibition (TWI). The experiments performed on mice with intracranial U373MG-LUC xenografts confirmed the efficacy of NPs-ZOL-Tf. These effects were paralleled by a higher intratumour localization of fluorescently-labeled-NPs-Tf both in intramuscular and intracranial xenografts. In conclusion, our results demonstrate that the encapsulation of ZOL increases the antitumor efficacy of this drug in glioblastoma through the acquisition of ability to cross the BBB.

Evaluation of estrogen receptor alpha activation by glyphosate-based herbicide constituents.

PubMed

Mesnage, Robin; Phedonos, Alexia; Biserni, Martina; Arno, Matthew; Balu, Sucharitha; Corton, J Christopher; Ugarte, Ricardo; Antoniou, Michael N

2017-10-01

The safety, including the endocrine disruptive capability, of glyphosate-based herbicides (GBHs) is a matter of intense debate. We evaluated the estrogenic potential of glyphosate, commercial GBHs and polyethoxylated tallowamine adjuvants present as co-formulants in GBHs. Glyphosate (≥10,000 μg/L or 59 μM) promoted proliferation of estrogen-dependent MCF-7 human breast cancer cells. Glyphosate also increased the expression of an estrogen response element-luciferase reporter gene (ERE-luc) in T47D-KBluc cells, which was blocked by the estrogen antagonist ICI 182,780. Commercial GBH formulations or their adjuvants alone did not exhibit estrogenic effects in either assay. Transcriptomics analysis of MCF-7 cells treated with glyphosate revealed changes in gene expression reflective of hormone-induced cell proliferation but did not overlap with an ERα gene expression biomarker. Calculation of glyphosate binding energy to ERα predicts a weak and unstable interaction (-4.10 kcal mol -1 ) compared to estradiol (-25.79 kcal mol -1 ), which suggests that activation of this receptor by glyphosate is via a ligand-independent mechanism. Induction of ERE-luc expression by the PKA signalling activator IBMX shows that ERE-luc is responsive to ligand-independent activation, suggesting a possible mechanism of glyphosate-mediated activation. Our study reveals that glyphosate, but not other components present in GBHs, can activate ERα in vitro, albeit at relatively high concentrations. Copyright © 2017 The Authors. Published by Elsevier Ltd.. All rights reserved.

Characterization of various promoter regions of the human DNA helicase-encoding genes and identification of duplicated ets (GGAA) motifs as an essential transcription regulatory element.

PubMed

Uchiumi, Fumiaki; Watanabe, Takeshi; Tanuma, Sei-ichi

2010-05-15

DNA helicases are important in the regulation of DNA transaction and thereby various cellular functions. In this study, we developed a cost-effective multiple DNA transfection assay with DEAE-dextran reagent and analyzed the promoter activities of the human DNA helicases. The 5'-flanking regions of the human DNA helicase-encoding genes were isolated and subcloned into luciferase (Luc) expression plasmids. They were coated onto 96-well plate and used for co-transfection with a renilla-Luc expression vector into various cells, and dual-Luc assays were performed. The profiles of promoter activities were dependent on cell lines used. Among these human DNA helicase genes, XPB, RecQL5, and RTEL promoters were activated during TPA-induced HL-60 cell differentiation. Interestingly, duplicated ets (GGAA) elements are commonly located around the transcription start sites of these genes. The duplicated GGAA motifs are also found in the promoters of DNA replication/repair synthesis factor genes including PARG, ATR, TERC, and Rb1. Mutation analyses suggested that the duplicated GGAA-motifs are necessary for the basal promoter activity in various cells and some of them positively respond to TPA in HL-60 cells. TPA-induced response of 44-bp in the RTEL promoter was attenuated by co-transfection of the PU.1 expression vector. These findings suggest that the duplicated ets motifs regulate DNA-repair associated gene expressions during macrophage-like differentiation of HL-60 cells. Copyright 2010 Elsevier Inc. All rights reserved.

Transgenic mice that accept Luciferase- or GFP-expressing syngeneic tumor cells at high efficiencies.

PubMed

Aoyama, Naoki; Miyoshi, Hiroyuki; Miyachi, Hitoshi; Sonoshita, Masahiro; Okabe, Masaru; Taketo, Makoto Mark

2018-05-11

Jellyfish green fluorescent protein (GFP) and firefly luciferase can serve as versatile tracking markers for identification and quantification of transplanted cancer cells in vivo. However, immune reactions against these markers can hamper the formation of syngraft tumors and metastasis that follows. Here, we report two transgenic (Tg) mouse lines that express nonfunctional mutant marker proteins, namely modified firefly luciferase (Luc2) or enhanced GFP (EGFP). These mice, named as Tg-mLuc2 and Tg-mEGFP, turned out to be immunologically tolerant to the respective tracking markers and thus efficiently accepted syngeneic cancer cells expressing the active forms of the markers. We then injected intrarectally the F 1 hybrid Tg mice (BALB/c × C57BL/6J) with Colon-26 (C26) colon cancer cells that originated from a BALB/c mouse. Even when C26 cells expressed active Luc2 or EGFP, they formed primary tumors in the Tg mice with only 10 4 cells per mouse compared with more than 10 6 cells required in the nontransgenic BALB/c hosts. Furthermore, we detected metastatic foci of C26 cells in the liver and lungs of the Tg mice by tracking the specific reporter activities. These results show the usefulness of the Tg mouse lines as recipients for transplantation experiments with the non-self tracking marker-expressing cells. © 2018 Molecular Biology Society of Japan and John Wiley & Sons Australia, Ltd.

Medical treatment of orthotopic glioblastoma with transferrin-conjugated nanoparticles encapsulating zoledronic acid

PubMed Central

Porru, Manuela; Zappavigna, Silvia; Salzano, Giuseppina; Luce, Amalia; Stoppacciaro, Antonella; Balestrieri, Maria Luisa; Artuso, Simona; Lusa, Sara; De Rosa, Giuseppe; Leonetti, Carlo; Caraglia, Michele

2014-01-01

Glioblastomas are highly aggressive adult brain tumors with poor clinical outcome. In the central nervous system (CNS) the blood-brain barrier (BBB) is the most important limiting factor for both development of new drugs and drug delivery. Here, we propose a new strategy to treat glioblastoma based on transferrin (Tf)-targeted self-assembled nanoparticles (NPs) incorporating zoledronic acid (ZOL) (NPs-ZOL-Tf). NPs-ZOL-Tf have been assessed on the glioblastoma cell line U373MG-LUC that showed a refractoriness in vitro to temozolomide (TMZ) and fotemustine (FTM). NPs-ZOL-Tf treatment resulted in higher in vitro cytotoxic activity than free ZOL. However, the potentiation of anti-proliferative activity of NPs-ZOL-Tf was superimposable to that one induced by NPs-ZOL (not armed with Tf). On the other hand, NPs-ZOL-Tf showed a higher antitumor efficacy if compared with that one caused by NPs-ZOL in immunosuppressed mice intramuscularly bearing U373MG-LUC xenografts, inducing a significant tumor weight inhibition (TWI). The experiments performed on mice with intracranial U373MG-LUC xenografts confirmed the efficacy of NPs-ZOL-Tf. These effects were paralleled by a higher intratumour localization of fluorescently-labeled-NPs-Tf both in intramuscular and intracranial xenografts. In conclusion, our results demonstrate that the encapsulation of ZOL increases the antitumor efficacy of this drug in glioblastoma through the acquisition of ability to cross the BBB. PMID:25431953

SAC3B, a central component of the mRNA export complex TREX-2, is required for prevention of epigenetic gene silencing in Arabidopsis

PubMed Central

Yang, Yu; La, Honggui; Tang, Kai; Miki, Daisuke; Yang, Lan; Wang, Bangshing; Duan, Cheng-Guo; Nie, Wenfeng; Wang, Xingang; Wang, Siwen; Pan, Yufeng; Tran, Elizabeth J.; An, Lizhe; Zhang, Huiming; Zhu, Jian-Kang

2017-01-01

Epigenetic regulation is important for organismal development and response to the environment. Alteration in epigenetic status has been known mostly from the perspective of enzymatic actions of DNA methylation and/or histone modifications. In a genetic screen for cellular factors involved in preventing epigenetic silencing, we isolated an Arabidopsis mutant defective in SAC3B, a component of the conserved TREX-2 complex that couples mRNA transcription with nuleo-cytoplasmic export. Arabidopsis SAC3B dysfunction causes gene silencing at transgenic and endogenous loci, accompanied by elevation in the repressive histone mark H3K9me2 and by reduction in RNA polymerase Pol II occupancy. SAC3B dysfunction does not alter promoter DNA methylation level of the transgene d35S::LUC, although the DNA demethylase ROS1 is also required for d35S::LUC anti-silencing. THP1 and NUA were identified as SAC3B-associated proteins whose mutations also caused d35S::LUC silencing. RNA-DNA hybrid exists at the repressed loci but is unrelated to gene suppression by the sac3b mutation. Genome-wide analyses demonstrated minor but clear involvement of SAC3B in regulating siRNAs and DNA methylation, particularly at a group of TAS and TAS-like loci. Together our results revealed not only a critical role of mRNA-export factors in transcriptional anti-silencing but also the contribution of SAC3B in shaping plant epigenetic landscapes. PMID:27672037

Ultrasound-induced hyperthermia for the spatio-temporal control of gene expression in bone repair

NASA Astrophysics Data System (ADS)

Wilson, Christopher; Padilla, Frédéric; Zhang, Man; Vilaboa, Nuria; Kripfgans, Oliver; Fowlkes, Brian; Franceschi, Renny

2012-10-01

Spatial and temporal control over the expression of growth/differentiation factors is of great interest for regeneration of bone, but technologies capable of providing tight and active control over gene expression remain elusive. We propose the use of focused ultrasound for the targeted activation of heat shock-sensitive expression systems in engineered bone. We report in vitro results with cells that express firefly luciferase (fLuc) under the control of a heat shock protein promoter. Cells were embedded in fibrin scaffolds and exposed to focused ultrasound, using a custom 3.3MHz transducer (focal length 4", f-number 1.33", focal dimension 1.2mm lateral FWHM) in CW mode for 2-20 minutes at intensities ISPTA=120-440 W/cm2. The kinetics of ultrasound-mediated activation of the cells was compared with that of strictly thermal activation. Bioluminescence imaging revealed fLuc expression in an area ≥2.5mm in diameter at the position of the ultrasound focus, and the diameter and intensity of the signal increased with the amplitude of the acoustic energy. We also found that ultrasound activated fLuc expression with substantially shorter exposures than thermal activation. Our results demonstrate the potential for focused ultrasound to selectively activate the expression of a gene of interest in an engineered tissue and suggest that focused ultrasound activates the heat shock pathway by a combination of thermal and non-thermal mechanisms.

Boron nitride nanotubes for gene silencing.

PubMed

Şen, Özlem; Çobandede, Zehra; Emanet, Melis; Bayrak, Ömer Faruk; Çulha, Mustafa

2017-09-01

Non-viral gene delivery is increasingly investigated as an alternative to viral vectors due to low toxicity and immunogenicity, easy preparation, tissue specificity, and ability to transfer larger sizes of genes. In this study, boron nitride nanotubes (BNNTs) are functionalized with oligonucleotides (oligo-BNNTs). The morpholinos complementary to the oligonucleotides attached to the BNNTs (morpholino/oligo-BNNTs) are hybridized to silence the luciferase gene. The morpholino/oligo-BNNTs conjugates are administered to luciferase-expressing cells (MDA-MB-231-luc2) and the luciferase activity is monitored. The luciferase activity is decreased when MDA-MB-231-luc2 cells were treated with morpholino/oligo-BNNTs. The study suggests that BNNTs can be used as a potential vector to transfect cells. BNNTs are potential new nanocarriers for gene delivery applications. Copyright © 2017 Elsevier B.V. All rights reserved.

SpyTag/SpyCatcher Cyclization Enhances the Thermostability of Firefly Luciferase

PubMed Central

Si, Meng; Xu, Qing

2016-01-01

SpyTag can spontaneously form a covalent isopeptide bond with its protein partner SpyCatcher. Firefly luciferase from Photinus pyralis was cyclized in vivo by fusing SpyCatcher at the N terminus and SpyTag at the C terminus. Circular LUC was more thermostable and alkali-tolerant than the wild type, without compromising the specific activity. Structural analysis indicated that the cyclized LUC increased the thermodynamic stability of the structure and remained more properly folded at high temperatures when compared with the wild type. We also prepared an N-terminally and C-terminally shortened form of the SpyCatcher protein and cyclization using this truncated form led to even more thermostability than the original form. Our findings suggest that cyclization with SpyTag and SpyCatcher is a promising and effective strategy to enhance thermostability of enzymes. PMID:27658030

GRC-2011-C-05116

NASA Image and Video Library

2009-01-24

Janis Niedra (RPT/Thermal Energy Conversion Branch) is using a Helmholtz coil to measure the approximate remanence of a magnet segment. The magnet segment being measured is similar to the ones used in the linear alternator of the Advanced Stirling Convertor (ASC) which was designed and built by Sunpower, Inc. The magnet (not shown in this view) is positioned in the slot cut along the top of the aluminum block which is positioned at the center of the coil in the region of uniform sensitivity. The ASC is the type of free-piston Stirling convertor that is used in the Advanced Stirling Radioisotope Generator (ASRG) currently being developed by Lockheed-Martin for the Department of Energy and NASA.

Afrotropical Melambiina: revision of the genus Tragardhus Koch, 1956 and redescription of Pseudemmallus Koch, 1956 (Tenebrionidae: Pedinini).

PubMed

Kamiński, Marcin Jan

2017-11-10

The genus Tragardhus Koch, 1956 (Pedinini: Melambiina) is revised to include nine Afrotropical species, five of which are new. A taxonomic treatment of the genus is provided including a morphological study, new species descriptions, keys, illustrations, and notes on species distributions. The following species are included: Tragardhus (Mitragardhus) nodosus Koch, 1956, T. (M.) ewae sp. nov., T. (M.) ryszardi sp. nov., T. (M.) zuzannae sp. nov., Tragardhus (Tragardhus) biapicalis Koch, 1956, T. (T.) glandipleurum Koch, 1956, T. (T.) jani sp. nov., T. (T.) majae sp. nov. and T. (T.) stigmaticus Koch, 1956. Additionally, a monotypic genus Pseudemmallus Koch, 1956, previously known from a single specimen representing P. aspericollis Koch, 1956, is redescribed based on newly available material.

Live imaging of the innate immune response in neonates reveals differential TLR2 dependent activation patterns in sterile inflammation and infection.

PubMed

Lalancette-Hébert, Melanie; Faustino, Joel; Thammisetty, Sai Sampath; Chip, Sophorn; Vexler, Zinaida S; Kriz, Jasna

2017-10-01

Activation of microglial cells in response to brain injury and/or immune stimuli is associated with a marked induction of Toll-like receptors (TLRs). While in adult brain, the contribution of individual TLRs, including TLR2, in pathophysiological cascades has been well established, their role and spatial and temporal induction patterns in immature brain are far less understood. To examine whether infectious stimuli and sterile inflammatory stimuli trigger distinct TLR2-mediated innate immune responses, we used three models in postnatal day 9 (P9) mice, a model of infection induced by systemic endotoxin injection and two models of sterile inflammation, intra-cortical IL-1β injection and transient middle cerebral artery occlusion (tMCAO). We took advantage of a transgenic mouse model bearing the dual reporter system luciferase/GFP under transcriptional control of a murine TLR2 promoter (TLR2-luc-GFP) to visualize the TLR2 response in the living neonatal brain and then determined neuroinflammation, microglial activation and leukocyte infiltration. We show that in physiological postnatal brain development the in vivo TLR2-luc signal undergoes a marked ∼30-fold decline and temporal-spatial changes during the second and third postnatal weeks. We then show that while endotoxin robustly induces the in vivo TLR2-luc signal in the living brain and increases levels of several inflammatory cytokines and chemokines, the in vivo TLR2-luc signal is reduced after both IL-1β and tMCAO and the inflammatory response is muted. Immunofluorescence revealed that microglial cells are the predominant source of TLR2 production during postnatal brain development and in all three neonatal models studied. Flow cytometry revealed developmental changes in CD11b + /CD45 + and CD11b + /Ly6C + cell populations, involvement of cells of the monocyte lineage, but lack of Ly6G + neutrophils or CD3 + cells in acutely injured neonatal brains. Cumulatively, our results suggest distinct TLR2 induction

A Small Molecule Inhibitor of ETV1, YK-4-279, Prevents Prostate Cancer Growth and Metastasis in a Mouse Xenograft Model

PubMed Central

Rahim, Said; Minas, Tsion; Hong, Sung-Hyeok; Justvig, Sarah; Çelik, Haydar; Kont, Yasemin Saygideger; Han, Jenny; Kallarakal, Abraham T.; Kong, Yali; Rudek, Michelle A.; Brown, Milton L.; Kallakury, Bhaskar; Toretsky, Jeffrey A.; Üren, Aykut

2014-01-01

Background The erythroblastosis virus E26 transforming sequences (ETS) family of transcription factors consists of a highly conserved group of genes that play important roles in cellular proliferation, differentiation, migration and invasion. Chromosomal translocations fusing ETS factors to promoters of androgen responsive genes have been found in prostate cancers, including the most clinically aggressive forms. ERG and ETV1 are the most commonly translocated ETS proteins. Over-expression of these proteins in prostate cancer cells results in a more invasive phenotype. Inhibition of ETS activity by small molecule inhibitors may provide a novel method for the treatment of prostate cancer. Methods and Findings We recently demonstrated that the small molecule YK-4-279 inhibits biological activity of ETV1 in fusion-positive prostate cancer cells leading to decreased motility and invasion in-vitro. Here, we present data from an in-vivo mouse xenograft model. SCID-beige mice were subcutaneously implanted with fusion-positive LNCaP-luc-M6 and fusion-negative PC-3M-luc-C6 tumors. Animals were treated with YK-4-279, and its effects on primary tumor growth and lung metastasis were evaluated. YK-4-279 treatment resulted in decreased growth of the primary tumor only in LNCaP-luc-M6 cohort. When primary tumors were grown to comparable sizes, YK-4-279 inhibited tumor metastasis to the lungs. Expression of ETV1 target genes MMP7, FKBP10 and GLYATL2 were reduced in YK-4-279 treated animals. ETS fusion-negative PC-3M-luc-C6 xenografts were unresponsive to the compound. Furthermore, YK-4-279 is a chiral molecule that exists as a racemic mixture of R and S enantiomers. We established that (S)-YK-4-279 is the active enantiomer in prostate cancer cells. Conclusion Our results demonstrate that YK-4-279 is a potent inhibitor of ETV1 and inhibits both the primary tumor growth and metastasis of fusion positive prostate cancer xenografts. Therefore, YK-4-279 or similar compounds may be

Evaluation of five dry particle deposition parameterizations for incorporation into atmospheric transport models

NASA Astrophysics Data System (ADS)

Khan, Tanvir R.; Perlinger, Judith A.

2017-10-01

Despite considerable effort to develop mechanistic dry particle deposition parameterizations for atmospheric transport models, current knowledge has been inadequate to propose quantitative measures of the relative performance of available parameterizations. In this study, we evaluated the performance of five dry particle deposition parameterizations developed by Zhang et al. (2001) (Z01), Petroff and Zhang (2010) (PZ10), Kouznetsov and Sofiev (2012) (KS12), Zhang and He (2014) (ZH14), and Zhang and Shao (2014) (ZS14), respectively. The evaluation was performed in three dimensions: model ability to reproduce observed deposition velocities, Vd (accuracy); the influence of imprecision in input parameter values on the modeled Vd (uncertainty); and identification of the most influential parameter(s) (sensitivity). The accuracy of the modeled Vd was evaluated using observations obtained from five land use categories (LUCs): grass, coniferous and deciduous forests, natural water, and ice/snow. To ascertain the uncertainty in modeled Vd, and quantify the influence of imprecision in key model input parameters, a Monte Carlo uncertainty analysis was performed. The Sobol' sensitivity analysis was conducted with the objective to determine the parameter ranking from the most to the least influential. Comparing the normalized mean bias factors (indicators of accuracy), we find that the ZH14 parameterization is the most accurate for all LUCs except for coniferous forest, for which it is second most accurate. From Monte Carlo simulations, the estimated mean normalized uncertainties in the modeled Vd obtained for seven particle sizes (ranging from 0.005 to 2.5 µm) for the five LUCs are 17, 12, 13, 16, and 27 % for the Z01, PZ10, KS12, ZH14, and ZS14 parameterizations, respectively. From the Sobol' sensitivity results, we suggest that the parameter rankings vary by particle size and LUC for a given parameterization. Overall, for dp = 0.001 to 1.0 µm, friction velocity was one of

INNOVATIVE BIODIESEL PRODUCTION: A SOLUTION TO THE SCIENTIFIC, TECHNICAL, AND EDUCATIONAL CHALLENGES OF SUSTAINABILITY

EPA Science Inventory

Loyola's STEP students completed over 20 team projects: Developed a business plan for biodiesel production, created the LUC biodiesel website, created the Bio­shorts documentaries, tabled at environmental events, publicized and put on two Biodiesel Forums (2nd one pending,...

Texas trip generation manual : 1st edition-volume 1 : user's guide.

DOT National Transportation Integrated Search

2014-08-01

The purpose of this Manual is to provide a summary of Texas trip generation data for various : Land Use Codes (LUCs) and time periods, for data obtained from workplace and special : generator (WSG) surveys performed as part of the Texas Travel Survey...

Synthesis of N-peptide-6-amino-D-luciferin Conjugates.

PubMed

Kovács, Anita K; Hegyes, Péter; Szebeni, Gábor J; Nagy, Lajos I; Puskás, László G; Tóth, Gábor K

2018-01-01

A general strategy for the synthesis of N -peptide-6-amino-D-luciferin conjugates has been developed. The applicability of the strategy was demonstrated with the preparation of a known substrate, N -Z-Asp-Glu-Val-Asp-6-amino-D-luciferin ( N -Z-DEVD-aLuc). N -Z-DEVD-aLuc was obtained via a hybrid liquid/solid phase synthesis method, in which the appropriately protected C-terminal amino acid was coupled to 6-amino-2-cyanobenzothiazole and the resulting conjugate was reacted with D-cysteine in order to get the protected amino acid-6-amino-D-luciferin conjugate, which was then attached to resin. The resulting loaded resin was used for the solid-phase synthesis of the desired N -peptide-6-amino-D-luciferin conjugate without difficulties, which was then attested with NMR spectroscopy and LC-MS, and successfully tested in a bioluminescent system.

Synthesis of N-peptide-6-amino-D-luciferin Conjugates

PubMed Central

Kovács, Anita K.; Hegyes, Péter; Szebeni, Gábor J.; Nagy, Lajos I.; Puskás, László G.; Tóth, Gábor K.

2018-01-01

A general strategy for the synthesis of N-peptide-6-amino-D-luciferin conjugates has been developed. The applicability of the strategy was demonstrated with the preparation of a known substrate, N-Z-Asp-Glu-Val-Asp-6-amino-D-luciferin (N-Z-DEVD-aLuc). N-Z-DEVD-aLuc was obtained via a hybrid liquid/solid phase synthesis method, in which the appropriately protected C-terminal amino acid was coupled to 6-amino-2-cyanobenzothiazole and the resulting conjugate was reacted with D-cysteine in order to get the protected amino acid-6-amino-D-luciferin conjugate, which was then attached to resin. The resulting loaded resin was used for the solid-phase synthesis of the desired N-peptide-6-amino-D-luciferin conjugate without difficulties, which was then attested with NMR spectroscopy and LC-MS, and successfully tested in a bioluminescent system. PMID:29725588

High field gradient targeting of magnetic nanoparticle-loaded endothelial cells to the surfaces of steel stents

PubMed Central

Polyak, Boris; Fishbein, Ilia; Chorny, Michael; Alferiev, Ivan; Williams, Darryl; Yellen, Ben; Friedman, Gary; Levy, Robert J.

2008-01-01

A cell delivery strategy was investigated that was hypothesized to enable magnetic targeting of endothelial cells to the steel surfaces of intraarterial stents because of the following mechanisms: (i) preloading cells with biodegradable polymeric superparamagnetic nanoparticles (MNPs), thereby rendering the cells magnetically responsive; and (ii) the induction of both magnetic field gradients around the wires of a steel stent and magnetic moments within MNPs because of a uniform external magnetic field, thereby targeting MNP-laden cells to the stent wires. In vitro studies demonstrated that MNP-loaded bovine aortic endothelial cells (BAECs) could be magnetically targeted to steel stent wires. In vivo MNP-loaded BAECs transduced with adenoviruses expressing luciferase (Luc) were targeted to stents deployed in rat carotid arteries in the presence of a uniform magnetic field with significantly greater Luc expression, detected by in vivo optical imaging, than nonmagnetic controls. PMID:18182491

Desertification, land use, and the transformation of global drylands

USGS Publications Warehouse

Bestelmeyer, Brandon T.; Okin, Gregory S.; Duniway, Michael C.; Archer, Steven R.; Sayre, Nathan F.; Williamson, Jebediah C.; Herrick, Jeffrey E.

2015-01-01

Desertification is an escalating concern in global drylands, yet assessments to guide management and policy responses are limited by ambiguity concerning the definition of “desertification” and what processes are involved. To improve clarity, we propose that assessments of desertification and land transformation be placed within a state change–land-use change (SC–LUC) framework. This framework considers desertification as state changes occurring within the context of particular land uses (eg rangeland, cropland) that interact with land-use change. State changes that can be readily reversed are distinguished from regime shifts, which are state changes involving persistent alterations to vegetation or soil properties. Pressures driving the transformation of rangelands to other types of land uses may be low, fluctuating, or high, and may influence and be influenced by state change. We discuss how the SC–LUC perspective can guide more effective assessment of desertification and management of drylands.

Combining Optical Reporter Proteins with Different Half-lives to Detect Temporal Evolution of Hypoxia and Reoxygenation in Tumors

PubMed Central

Danhier, Pierre; Krishnamachary, Balaji; Bharti, Santosh; Kakkad, Samata; Mironchik, Yelena; Bhujwalla, Zaver M.

2015-01-01

Here we have developed a hypoxia response element driven imaging strategy that combined the hypoxia-driven expression of two optical reporters with different half-lives to detect temporal changes in hypoxia and hypoxia inducible factor (HIF) activity. For this purpose, human prostate cancer PC3 cells were transfected with the luciferase gene fused with an oxygen-dependent degradation domain (ODD-luc) and a variant of the enhanced green fluorescent protein (EGFP). Both ODD-luciferase and EGFP were under the promotion of a poly-hypoxia-response element sequence (5xHRE). The cells constitutively expressed tdTomato red fluorescent protein. For validating the imaging strategy, cells were incubated under hypoxia (1% O2) for 48 hours and then reoxygenated. The luciferase activity of PC3-HRE-EGFP/HRE-ODD-luc/tdtomato cells detected by bioluminescent imaging rapidly decreased after reoxygenation, whereas EGFP levels in these cells remained stable for several hours. After in vitro validation, PC3-HRE-EGFP/HRE-ODD-luc/tdtomato tumors were implanted subcutaneously and orthotopically in nude male mice and imaged in vivo and ex vivo using optical imaging in proof-of-principle studies to demonstrate differences in optical patterns between EGFP expression and bioluminescence. This novel "timer" imaging strategy of combining the short-lived ODD-luciferase and the long-lived EGFP can provide a time frame of HRE activation in PC3 prostate cancer cells and will be useful to understand the temporal changes in hypoxia and HIF activity during cancer progression and following treatments including HIF targeting strategies. PMID:26696369

Raman spectroscopy of bone metastasis

NASA Astrophysics Data System (ADS)

Esmonde-White, Karen A.; Sottnik, Joseph; Morris, Michael; Keller, Evan

2012-02-01

Raman spectroscopy of bone has been used to characterize chemical changes occurring in diseases such as osteoporosis, osteoarthritis and osteomyelitis. Metastasis of cancer into bone causes changes to bone quality that are similar to those observed in osteoporosis, such as decreased bone strength, but with an accelerated timeframe. In particular, osteolytic (bone degrading) lesions in bone metastasis have a marked effect on patient quality of life because of increased risk of fractures, pain, and hypercalcemia. We use Raman spectroscopy to examine bone from two different mouse models of osteolytic bone metastasis. Raman spectroscopy measures physicochemical information which cannot be obtained through standard biochemical and histological measurements. This study was reviewed and approved by the University of Michigan University Committee on the Care and Use of Animals. Two mouse models of prostate cancer bone metastasis, RM1 (n=3) and PC3-luc (n=4) were examined. Tibiae were injected with RM1 or PC3-luc cancer cells, while the contralateral tibiae received a placebo injection for use as controls. After 2 weeks of incubation, the mice were sacrificed and the tibiae were examined by Raman microspectroscopy (λ=785 nm). Spectroscopic markers corresponding to mineral stoichiometry, bone mineralization, and mineral crystallinity were compared in spectra from the cancerous and control tibiae. X-ray imaging of the tibia confirmed extensive osteolysis in the RM1 mice, with tumor invasion into adjoining soft tissue and moderate osteolysis in the PC3-luc mice. Raman spectroscopic markers indicate that osteolytic lesions are less mineralized than normal bone tissue, with an altered mineral stoichiometry and crystallinity.

An inducible model of abacterial prostatitis induces antigen specific inflammatory and proliferative changes in the murine prostate

PubMed Central

Haverkamp, Jessica M.; Charbonneau, Bridget; Meyerholz, David K.; Cohen, Michael B.; Snyder, Paul W.; Svensson, Robert U.; Henry, Michael D.; Wang, Hsing- Hui

2011-01-01

Background Prostatitis is a poorly understood disease and increasing evidence suggests inflammation is involved in other prostatic diseases, including prostate cancer. Methods The ability of pre-activated CD8 T cells to induce prostatitis was examined by adoptive transfer into POET-3 mice or POET-3/Luc/Pten−/+ mice. Characterization of the inflammatory response was determined by examining leukocyte infiltration by histological analysis, flow cytometry and by evaluating cytokine and chemokine levels in prostate tissue. The impact of inflammation on the prostate was evaluated by monitoring epithelial cell proliferation over time. Results Initiation of inflammation by ovalbumin specific CD8+ T cells (OT-I cells) resulted in development of acute prostatitis in the anterior, dorsolateral and anterior prostate of POET-3 and POET-3/Luc/Pten−/+ mice. Acute prostatitis was characterized by recruitment of adoptively transferred OT-I cells and importantly, autologous CD4+ and CD8+ T cells, myeloid-derived suppressor cells (MDSC) and regulatory T cells (Treg). In concert with leukocyte infiltration elevated levels of pro-inflammatory cytokines and chemokines were observed. Inflammation also resulted in marked epithelial cell proliferation that was sustained up to 80 days post adoptive-transfer of OT-I cells. Conclusions The POET-3 model represents a novel mouse model to study both acute and chronic prostate inflammation in an antigen-specific system. Further, the POET-3 mouse model can be crossed with other genetic models of disease such as the C57/Luc/Pten−/− model of prostate cancer, allowing the impact of prostatitis on other prostatic diseases to be evaluated. PMID:21656824

Direct comparison of administration routes for AAV8-mediated ocular gene therapy.

PubMed

Igarashi, Tsutomu; Miyake, Koichi; Asakawa, Nagisa; Miyake, Noriko; Shimada, Takashi; Takahashi, Hiroshi

2013-05-01

We recently demonstrated that direct subretinal (SR) injection of adeno-associated virus (AAV) type 8 (AAV8) into photoreceptor cells and retinal pigment epithelium (RPE) is a highly efficient model of gene delivery. The current study compared transduction efficiency and expression patterns associated with various routes of vector administration. The efficacy of intravitreal (VT), SR and subconjunctival (SC) injections for delivery of AAV8-derived vectors, i.e. those expressing luciferase (Luc) and enhanced green fluorescent protein (GFP) - AAV8/Luc and AAV8/GFP, respectively - were compared in an animal (mouse) model (n = 8 mice/group). Transduction efficiency and expression patterns were examined at post-injection weeks 1 and 2, and months 1, 3, 6 and 12 via in vivo imaging. One year after AAV injection, AAV8/Luc-treated mice exhibited stable and sustained high expression of vector in the VT and SR groups, but not in the SC group (VT:SR:SC = 3,218:2,923:115; 1 × 10(5 )photons/s). Histological analysis showed that GFP expression was observed in the inner retina of VT group mice, and in photoreceptor cells and RPE of SR group mice, whereas no GFP expression was noted in the SC group. Electroretinography (ERG) revealed adverse effects following SR delivery. Results suggest that both SR and VT injections of AAV8 vectors are useful routes for administering ocular gene therapy, and stress the importance of selecting an appropriate administration route, i.e. one that targets specific cells, for treating ocular disorders.

Sibling Communication in "Star Trek: The Next Generation": Conflicts between Brothers.

ERIC Educational Resources Information Center

Fuss-Reineck, Marilyn

"Star Trek: The Next Generation" depicts sibling relationships between Data and Lore (android), Worf and Kurn (Klingon) and Jean-Luc and Robert (human) that illustrate conflictual communication and suggest teaching applications. The most disturbing empirical communication research on conflict between brothers reports that male sibling…

Materials Data on LuC2 (SG:139) by Materials Project

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kristin Persson

2014-07-09

Computed materials data using density functional theory calculations. These calculations determine the electronic structure of bulk materials by solving approximations to the Schrodinger equation. For more information, see https://materialsproject.org/docs/calculations

77 FR 8258 - Availability of ICCVAM Evaluation Report and Recommendations on the Usefulness and Limitations of...

Federal Register 2010, 2011, 2012, 2013, 2014

2012-02-14

... development of BG1Luc ER TA test method performance standards. ICCVAM assigned the activities a high priority... Vitro Test Methods for Detecting Potential Endocrine Disruptors. Research Triangle Park, NC: National...Final.pdf . ICCVAM. 2003a. ICCVAM Evaluation of In Vitro Test Methods For Detecting Potential Endocrine...

IN VITRO TO IN VIVO SCREENING OF THYROID HORMONE RECEPTOR DISRUPTING CHEMICALS

EPA Science Inventory

Upon completion of these studies, we will have established the predictive value of the GH3.TRE-LUC cell line to detect chemicals that can impact TH regulated gene expression and TH regulated developmental events in vivo. These studies have excellent potential to discover new c...

From carbonate platform to euxinic sea - the collapse of an Early/Middle Devonian reef, Cantabrian Mountains (Spain)

NASA Astrophysics Data System (ADS)

Loevezijn, Gerard B. S. van; Raven, J. G. M.

2017-12-01

The Santa Lucía Formation represents the major phase in Devonian reef development of the Cantabrian Zone (Cantabrian Mountains, northwest Spain). In the present study the transition from the carbonate platform deposits of the Santa Lucía Formation to the overlying euxinic basinal deposits of the Huergas Formation is described. These transitional strata are connected to the Basal Choteč Event and represent a condensed sedimentation of micritic dark-grey and black limestones with an upward increase of dark shale intercalations with iron mineralisation surfaces and storm-induced brachiopod coquinas. The transitional beds are grouped into a new unit, the Cabornera Bed, which consists of limestone, limestone-shale and shale facies associations, representing a sediment-starved euxinic offshore area just below the storm wave base. Four stages in reef decline can be recognised: a reef stage, an oxygen-depleted, nutrient-rich stage, a siliciclastic-influx stage and a pelagic-siliciclastic stage. Additional geochemical and geophysical investigations are needed to verify the results presented herein.

Highly specific expression of luciferase gene in lungs of naive nude mice directed by prostate-specific antigen promoter

DOE Office of Scientific and Technical Information (OSTI.GOV)

Li Hongwei; Department of Neurological Surgery, University of Virginia Health System, Charlottesville, VA 22908; Li Jinzhong

PSA promoter has been demonstrated the utility for tissue-specific toxic gene therapy in prostate cancer models. Characterization of foreign gene overexpression in normal animals elicited by PSA promoter should help evaluate therapy safety. Here we constructed an adenovirus vector (AdPSA-Luc), containing firefly luciferase gene under the control of the 5837 bp long prostate-specific antigen promoter. A charge coupled device video camera was used to non-invasively image expression of firefly luciferase in nude mice on days 3, 7, 11 after injection of 2 x 10{sup 9} PFU of AdPSA-Luc virus via tail vein. The result showed highly specific expression of themore » luciferase gene in lungs of mice from day 7. The finding indicates the potential limitations of the suicide gene therapy of prostate cancer based on selectivity of PSA promoter. By contrary, it has encouraging implications for further development of vectors via PSA promoter to enable gene therapy for pulmonary diseases.« less

Generation of iPS-derived model cells for analyses of hair shaft differentiation.

PubMed

Kido, Takumi; Horigome, Tomoatsu; Uda, Minori; Adachi, Naoki; Hirai, Yohei

2017-09-01

Biological evaluation of hair growth/differentiation activity in vitro has been a formidable challenge, primarily due to the lack of relevant model cell systems. To solve this problem, we generated a stable model cell line in which successive differentiation via epidermal progenitors to hair components is easily inducible and traceable. Mouse induced pluripotent stem (iPS) cell-derived cells were selected to stably express a tetracycline (Tet)-inducible bone morphogenic protein-4 (BMP4) expression cassette and a luciferase reporter driven by a hair-specific keratin 31 gene (krt31) promoter (Tet-BMP4-KRT31-Luc iPS). While Tet- BMP4-KRT31-Luc iPS cells could be maintained as stable iPS cells, the cells differentiated to produce luciferase luminescence in the presence of all-trans retinoic acid (RA) and doxycycline (Dox), and addition of a hair differentiation factor significantly increased luciferase fluorescence. Thus, this cell line may provide a reliable cell-based screening system to evaluate drug candidates for hair differentiation activity.

First principle study of electronic nanoscale structure of In x Ga1- x P with variable size, shape and alloying percentage

NASA Astrophysics Data System (ADS)

Hussein, M. T.; Kasim, T.; Abdulsattar, M. A.

2013-11-01

In present work, we investigate electronic properties of alloying percentage of In x Ga1- x P compound with different sizes of superlattice large unit cell (LUC) method with 8, 16, 54, and 64 nanocrystals core atoms. The size and type of alloying compound are varied so that it can be tuned to a required application. To determine properties of indium gallium phosphide nanocrystals density functional theory at the generalized-gradient approximation level coupled with LUC method is used to simulate electronic structure of zinc blende indium gallium phosphide nanocrystals that have dimensions around 2-2.8 nm. The calculated properties include lattice constant, energy gap, valence band width, cohesive energy, density of states (DOS) etc. Results show that laws that are applied at microscale alloying percentage are no more applicable at the present nanoscale. Results also show that size, shape and quantum effects are strong. Many properties fluctuate at nanoscale while others converge to definite values. DOS summarizes many of the above quantities.

Comparison of human optimized bacterial luciferase, firefly luciferase, and green fluorescent protein for continuous imaging of cell culture and animal models

NASA Astrophysics Data System (ADS)

Close, Dan M.; Hahn, Ruth E.; Patterson, Stacey S.; Baek, Seung J.; Ripp, Steven A.; Sayler, Gary S.

2011-04-01

Bioluminescent and fluorescent reporter systems have enabled the rapid and continued growth of the optical imaging field over the last two decades. Of particular interest has been noninvasive signal detection from mammalian tissues under both cell culture and whole animal settings. Here we report on the advantages and limitations of imaging using a recently introduced bacterial luciferase (lux) reporter system engineered for increased bioluminescent expression in the mammalian cellular environment. Comparison with the bioluminescent firefly luciferase (Luc) system and green fluorescent protein system under cell culture conditions demonstrated a reduced average radiance, but maintained a more constant level of bioluminescent output without the need for substrate addition or exogenous excitation to elicit the production of signal. Comparison with the Luc system following subcutaneous and intraperitoneal injection into nude mice hosts demonstrated the ability to obtain similar detection patterns with in vitro experiments at cell population sizes above 2.5 × 104 cells but at the cost of increasing overall image integration time.

Paddy rice productivity under climate and land-use change in northern Japan

NASA Astrophysics Data System (ADS)

Yoshida, R.; Fukui, S.; Shimada, T.; Hasegawa, T.; Iwasaki, T.

2013-12-01

An evaluation of the best rice cultivar under climate change is an important issue because the projected climate change has a potential to bring a negative impact on crop yield. In this study, we estimate an impact of climate change on rice yield and potential best cultivar in northern Japan where the larger paddy field is located than other regions in Japan. Two global climate model data, MIROC5 (RCP 4.5) and MRI-AGCM (SRES A1B), are applied as the future scenario. These data are too coarse to resolve the regional differences in northern Japan; we conduct the downscale experiments by a regional climate model (JMA-NHM) with a 10-km grid spacing. Considering that rice yield is sensitive to warm season climate, we conduct the downscaling from 28th May to 1st September during 1981-2000 and 2081-2099. The biases of downscaled two scenarios are corrected to match their cumulative distribution functions (CDF) of present climate with that of the station-based observation. The derived CDF-based biases are also used to correct the future scenarios. These corrected scenarios are applied to rice growth model (NIAES-Rice). To take account for the impacts of land use change (LUC) on climate and rice yield, we consider the additional temperature changes due to the LUC. As a reference, we add the LUC-induced temperature change in southwest Japan because the data are available from the previous study. We first check the climate change in northern Japan. General tendencies derived from the bias-corrected-downscaled future climates are that 1) surface warming was approximately twice in low elevation area relative to mountainous area and 2) downward shortwave radiation homogeneously increased 7-8 W m-2. Then, we evaluated the simulated yield through comparing with observation. Using observed ambient conditions as input data, the NIAES-Rice model provides the reasonable performance in simulating the rice yield with biases ranging from -19.0 to 29.2 % in prefecture base. Climate

Ar-Ar and U-Pb ages of marble-hosted ruby deposits from Central and South-east Asia

NASA Astrophysics Data System (ADS)

Garnier, V.; Giuliani, G.; Maluski, H.; Ohnenstetter, D.; Deloule, E.

2003-04-01

Marble-hosted ruby deposits represent the first source of gemstones in Asia. The deposits from Jegdalek (Afghanistan), Hunza Valley (Pakistan), Nangimali (Azad-Kashmir), Chumar, Ruyil (Nepal), Mogok (Myanmar), Luc Yen, Yen Bai and Quy Chau (Vietnam) were dated using the 40Ar-39Ar laser stepwise heating technique on syngenetic micas. The following ages were obtained : 24.7 ± 0.3 Ma at Jegdalek ; 10.8 ± 0.3 to 5.4 ± 0.3 Ma at Hunza ; 17.2 ± 0.2 to 15.3 ± 0.1 Ma at Nangimali ; 4.6 ± 0.1 Ma at Ruyil ; 5.6 ± 0.4 Ma at Chumar ; 18.7 ± 0.2 to 17.1 ± 0.2 Ma at Mogok ; 33.8 ± 0.4 to 30.8 ± 0.8 Ma at Luc Yen ; 24.4 ± 0.4 to 23.2 ± 0.6 Ma at Yen Bai, 22.1 ± 0.6 to 21.6 ± 0.7 Ma at Quy Chau. These ages represent cooling ages and thus minimum ages for ruby formation. The ages obtained for Nangimali are close to the Ar-Ar cooling age of 19 Ma recorded in the Chichi granite, North to the ruby deposit. However, (C,O)-isotopic studies of the ruby-bearing marbles show no genetic relation between granite emplacement and ruby deposition in this area. The age found at Jegdalek is similar to the K-Ar ages obtained on the Sairobi pegmatitic dykes (20-26 Ma) and of the Jalalabad pluton (25 Ma), located close to the ruby deposit. At Mogok, the ruby deposits yield ages close to those obtained on high grade metamorphic and foliated intrusive regional rocks (15.8 ± 0.7 - 19.5 ± 1.0 Ma). The ages obtained at Chumar and Ruyil agree with those of the Lesser Himalaya Formation (12 - 6 Ma). Those found at Quy Chau agree with those found for the shear zone activity. Furthermore, U-Pb dating was done on zircons included in a ruby from Luc Yen and spinels in marble from Luc Yen and Hunza. The wide range of 238U-206Pb ages obtained for Luc Yen (266 - 45 Ma) evidences a complex metamorphic history. Ruby crystallised at 45 Ma during ductile activity of the Red River shear zone. At Hunza, an 238U-206Pb age of 94.0 ± 2.1 Ma obtained on inherited zircons confirms the U-Pb age obtained on

Impossible Practice and Theories of the Impossible: A Response to Helene Illeris's "Potentials of Togetherness"

ERIC Educational Resources Information Center

Kallio-Tavin, Mira

2014-01-01

In a recent commentary in "Studies in Art Education," Helene Illeris (2013) discussed the idea of "performative experimental communities" via a critique of visual culture pedagogy and the romanticism of community-oriented art education in Nordic countries. Illeris underpinned her arguments with Jean-Luc Nancy's (1997)…

20180416 - Retrofitting an Estrogen Receptor Transactivation Assay with Metabolic Competence Using Alginate Immobilization of Metabolic Enzymes (AIME) (SETAC HTS)

EPA Science Inventory

The VM7Luc4E2 estrogen receptor (ER) transactivation assay is an OECD approved method (TG 457) for the detection of ER agonists and antagonists, and is also part of the Tox21 high-throughput screening (HTS) portfolio. Despite international acceptance as a screening assay, immorta...

20180312 - Retrofitting an Estrogen Receptor Transactivation Assay with Metabolic Competence Using Alginate Immobilization of Metabolic Enzymes (AIME) (SOT)

EPA Science Inventory

The VM7Luc4E2 estrogen receptor (ER) transactivation assay is an OECD approved method (TG 457) for the detection of ER agonists and antagonists, and is also part of the Tox21 high-throughput screening (HTS) portfolio. Despite its international acceptance as a screening assay, imm...

The New Spirit of Capitalism in European Liberal Arts Programs

ERIC Educational Resources Information Center

Claus, Jakob; Meckel, Thomas; Pätz, Farina

2018-01-01

The following paper suggests a connection between recent developments in the justification of the capitalist system and contemporary European Liberal Arts programs. By looking at Luc Boltanski's and Eve Chiapello's study on "The New Spirit Of Capitalism" and Gilles Deleuze's term of "societies of control" we highlight a pivot…

Soil organic carbon pool's contribution to climate change mitigation on marginal land of a Mediterranean montane area in Italy.

PubMed

Tommaso, Chiti; Emanuele, Blasi; Guido, Pellis; Lucia, Perugini; Vincenza, Chiriacò Maria; Riccardo, Valentini

2018-07-15

To evaluate the mitigation potential provided by the SOC pool, we investigated the impact of woody encroachment in the 0-30 cm depth of mineral soil across a natural succession from abandoned pastures and croplands to broadleaves forests on the central Apennine in Italy. In parallel, to assess the effect of the land use change (LUC) from cropland to pasture, a series of pastures established on former agricultural sites, abandoned at different time in the past, were also investigated. Our results show that woody encroachment on former pastures and croplands contributes largely to mitigate climate change, with an increase of the original SOC stock of 45% (40.5 Mg C ha -1 ) and 120% (66.5 Mg C ha -1 ), respectively. Also the LUC from croplands to pastures, greatly contributes to climate change mitigation trough a SOC increase of about 80% of the original SOC (45.9 Mg C ha -1 ). The management of abandoned lands represent a crucial point in the mitigation potential of agriculture and forestry activities, and particularly the role of the SOC pool. A policy effort should focus on minimizing the risk of speculative management options, particularly when the value of woody biomass become convenient to supply new energy systems allowing monetizing a long term forests productivity. In conclusion, despite both the land abandonment and the LUC can have a different impact on the SOC pool under different climatic conditions, these results can be useful to improve the SOC estimates in the National greenhouse gases Inventory at country level. Copyright © 2018 Elsevier Ltd. All rights reserved.

Preclinical Evaluation of the Supercritical Extract of Azadirachta Indica (Neem) Leaves In Vitro and In Vivo on Inhibition of Prostate Cancer Tumor Growth

PubMed Central

Wu, Qiang; Kohli, Manish; Bergen, H. Robert; Cheville, John C.; Karnes, R. Jeffrey; Cao, Hong; Young, Charles Y.F.; Tindall, Donald J.; McNiven, Mark A.; Donkena, Krishna Vanaja

2015-01-01

Azadirachta indica, commonly known as neem, has gained worldwide prominence because of its medical properties, namely antitumor, antiviral, anti-inflammatory, antihyperglycemic, antifungal, and antibacterial activities. Despite these promising results, gaps remain in our understanding of the molecular mechanism of action of neem compounds and their potential for use in clinical trials. We investigated supercritical extract of neem leaves (SENL) for the following: molecular targets in vitro, in vivo efficacy to inhibit tumor growth, and bioactive compounds that exert antitumor activity. Treatment of LNCaP-luc2 prostate cancer cells with SENL suppressed dihydrotestosterone-induced androgen receptor and prostate-specific antigen levels. SENL inhibited integrin β1, calreticulin, and focal adhesion kinase activation in LNCaP-luc2 and PC3 prostate cancer cells. Oral administration of SENL significantly reduced LNCaP-luc2 xenograft tumor growth in mice with the formation of hyalinized fibrous tumor tissue, reduction in the prostate-specific antigen, and increase in AKR1C2 levels. To identify the active anticancer compounds, we fractionated SENL by high-pressure liquid chromatography and evaluated 16 peaks for cytotoxic activity. Four of the 16 peaks exhibited significant cytotoxic activity against prostate cancer cells. Mass spectrometry of the isolated peaks suggested the compounds with cytotoxic activity were nimbandiol, nimbolide, 2′,3′-dihydronimbolide, and 28-deoxonim-bolide. Analysis of tumor tissue and plasma samples from mice treated with SENL indicated 28-deoxonim-bolide and nimbolide as the bioactive compounds. Overall, our data revealed the bioactive compounds in SENL and suggested that the anticancer activity could be mediated through alteration in androgen receptor and calreticulin levels in prostate cancer. PMID:24674886

Combining Optical Reporter Proteins with Different Half-lives to Detect Temporal Evolution of Hypoxia and Reoxygenation in Tumors.

PubMed

Danhier, Pierre; Krishnamachary, Balaji; Bharti, Santosh; Kakkad, Samata; Mironchik, Yelena; Bhujwalla, Zaver M

2015-12-01

Here we have developed a hypoxia response element driven imaging strategy that combined the hypoxia-driven expression of two optical reporters with different half-lives to detect temporal changes in hypoxia and hypoxia inducible factor (HIF) activity. For this purpose, human prostate cancer PC3 cells were transfected with the luciferase gene fused with an oxygen-dependent degradation domain (ODD-luc) and a variant of the enhanced green fluorescent protein (EGFP). Both ODD-luciferase and EGFP were under the promotion of a poly-hypoxia-response element sequence (5xHRE). The cells constitutively expressed tdTomato red fluorescent protein. For validating the imaging strategy, cells were incubated under hypoxia (1% O2) for 48 hours and then reoxygenated. The luciferase activity of PC3-HRE-EGFP/HRE-ODD-luc/tdtomato cells detected by bioluminescent imaging rapidly decreased after reoxygenation, whereas EGFP levels in these cells remained stable for several hours. After in vitro validation, PC3-HRE-EGFP/HRE-ODD-luc/tdtomato tumors were implanted subcutaneously and orthotopically in nude male mice and imaged in vivo and ex vivo using optical imaging in proof-of-principle studies to demonstrate differences in optical patterns between EGFP expression and bioluminescence. This novel "timer" imaging strategy of combining the short-lived ODD-luciferase and the long-lived EGFP can provide a time frame of HRE activation in PC3 prostate cancer cells and will be useful to understand the temporal changes in hypoxia and HIF activity during cancer progression and following treatments including HIF targeting strategies. Copyright © 2015 Nencki Institute of Experimental Biology, Polish Academy of Sciences,. Published by Elsevier Inc. All rights reserved.

Systemic Immunotherapy of Non–Muscle Invasive Mouse Bladder Cancer with Avelumab, an Anti–PD-L1 Immune Checkpoint Inhibitor

PubMed Central

Vandeveer, Amanda J.; Fallon, Jonathan K.; Tighe, Robert; Sabzevari, Helen; Schlom, Jeffrey; Greiner, John W.

2016-01-01

Bacillus Calmette-Guerin (BCG) is the standard of care for intravesical therapy for carcinoma in situ and non–muscle invasive, nonmetastatic human urothelial carcinoma. While the responsiveness to this immunotherapeutic is believed to be linked with (i) a high number of somatic mutations and (ii) a large number of tumor-infiltrating lymphocytes, recent findings of the roles that inhibitory immune receptors and their ligands play in tumor evasion may provide insights into the limitations of the effectiveness of BCG and offer new targets for immune-based therapy. In this study, an aggressive, bioluminescent orthotopic bladder cancer model, MB49 tumor cells transfected with luciferase (MB49luc), was used to study the antitumor effects of avelumab, an antibody to PD-L1. MB49luc murine tumor cells form multifocal tumors on the mucosal wall of the bladder reminiscent of non–muscle invasive, nonmetastatic urothelial carcinomas. MB49luc bladder tumors are highly positive for the expression of PD-L1 and avelumab administration induced significant (P<0.05) antitumor effects. These antitumor effects were more dependent on the presence of CD4 than CD8 T cells, as determined by in vivo immune cell depletions. The findings suggest that in this bladder tumor model, interruption of the immune suppressive PD-1/PD-L1 complex releases a local adaptive immune response that, in turn, reduces tumor growth. This bladder tumor model can be used to further identify host antitumor immune mechanisms and evaluate combinations of immune-based therapies for carcinoma in situ and non–muscle invasive, nonmetastatic urothelial carcinoma, to provide the rationale for subsequent clinical studies. PMID:26921031

Sonochemiluminescence of lucigenin: Evidence of superoxide radical anion formation by ultrasonic irradiation

NASA Astrophysics Data System (ADS)

Matsuoka, Masanori; Takahashi, Fumiki; Asakura, Yoshiyuki; Jin, Jiye

2016-07-01

The sonochemiluminescence (SCL) behavior of lucigenin (Luc2+) has been studied in aqueous solutions irradiated with 500 kHz ultrasound. Compared with the SCL of a luminol system, a tremendously increased SCL intensity is observed from 50 µM Luc2+ aqueous solution (pH =11) when small amounts of coreactants such as 2-propanol coexist. It is shown that SCL intensity strongly depends on the presence of dissolved gases such as air, O2, N2, and Ar. The highest SCL intensity is obtained in an O2-saturated solution, indicating that molecular oxygen is required to generate SCL. Since SCL intensity is quenched completely in the presence of superoxide dismutase (SOD), an enzyme that can catalyze the disproportionation of O2 •-, the generation of O2 •- in the ultrasonic reaction field is important in the SCL of Luc2+. In this work, the evidence of O2 •- production is examined by a spectrofluorometric method using 2-(2-pyridyl)benzothiazoline as the fluorescent probe. The results indicate that the yield of O2 •- is markedly increased in the O2-saturated solutions when a small amount of 2-propanol coexists, which is consistent with the results of SCL measurements. 2-Propanol in the interfacial region of a cavitation bubble reacts with a hydroxyl radical (•OH) to form a 2-propanol radical, CH3C•(OH)CH3, which can subsequently react with dissolved oxygen to generate O2 •-. The most likely pathways for SCL as well as the spatial distribution of SCL in a microreactor are discussed in this study.

Dissociation of Per1 and Bmal1 circadian rhythms in the suprachiasmatic nucleus in parallel with behavioral outputs

PubMed Central

Ono, Daisuke; Honma, Sato; Nakajima, Yoshihiro; Kuroda, Shigeru; Enoki, Ryosuke; Honma, Ken-ichi

2017-01-01

The temporal order of physiology and behavior in mammals is primarily regulated by the circadian pacemaker located in the hypothalamic suprachiasmatic nucleus (SCN). Taking advantage of bioluminescence reporters, we monitored the circadian rhythms of the expression of clock genes Per1 and Bmal1 in the SCN of freely moving mice and found that the rate of phase shifts induced by a single light pulse was different in the two rhythms. The Per1-luc rhythm was phase-delayed instantaneously by the light presented at the subjective evening in parallel with the activity onset of behavioral rhythm, whereas the Bmal1-ELuc rhythm was phase-delayed gradually, similar to the activity offset. The dissociation was confirmed in cultured SCN slices of mice carrying both Per1-luc and Bmal1-ELuc reporters. The two rhythms in a single SCN slice showed significantly different periods in a long-term (3 wk) culture and were internally desynchronized. Regional specificity in the SCN was not detected for the period of Per1-luc and Bmal1-ELuc rhythms. Furthermore, neither is synchronized with circadian intracellular Ca2+ rhythms monitored by a calcium indicator, GCaMP6s, or with firing rhythms monitored on a multielectrode array dish, although the coupling between the circadian firing and Ca2+ rhythms persisted during culture. These findings indicate that the expressions of two key clock genes, Per1 and Bmal1, in the SCN are regulated in such a way that they may adopt different phases and free-running periods relative to each other and are respectively associated with the expression of activity onset and offset. PMID:28416676

Breast Cancer Cell Colonization of the Human Bone Marrow Adipose Tissue Niche.

PubMed

Templeton, Zach S; Lie, Wen-Rong; Wang, Weiqi; Rosenberg-Hasson, Yael; Alluri, Rajiv V; Tamaresis, John S; Bachmann, Michael H; Lee, Kitty; Maloney, William J; Contag, Christopher H; King, Bonnie L

2015-12-01

Bone is a preferred site of breast cancer metastasis, suggesting the presence of tissue-specific features that attract and promote the outgrowth of breast cancer cells. We sought to identify parameters of human bone tissue associated with breast cancer cell osteotropism and colonization in the metastatic niche. Migration and colonization patterns of MDA-MB-231-fLuc-EGFP (luciferase-enhanced green fluorescence protein) and MCF-7-fLuc-EGFP breast cancer cells were studied in co-culture with cancellous bone tissue fragments isolated from 14 hip arthroplasties. Breast cancer cell migration into tissues and toward tissue-conditioned medium was measured in Transwell migration chambers using bioluminescence imaging and analyzed as a function of secreted factors measured by multiplex immunoassay. Patterns of breast cancer cell colonization were evaluated with fluorescence microscopy and immunohistochemistry. Enhanced MDA-MB-231-fLuc-EGFP breast cancer cell migration to bone-conditioned versus control medium was observed in 12/14 specimens (P = .0014) and correlated significantly with increasing levels of the adipokines/cytokines leptin (P = .006) and IL-1β (P = .001) in univariate and multivariate regression analyses. Fluorescence microscopy and immunohistochemistry of fragments underscored the extreme adiposity of adult human bone tissues and revealed extensive breast cancer cell colonization within the marrow adipose tissue compartment. Our results show that breast cancer cells migrate to human bone tissue-conditioned medium in association with increasing levels of leptin and IL-1β, and colonize the bone marrow adipose tissue compartment of cultured fragments. Bone marrow adipose tissue and its molecular signals may be important but understudied components of the breast cancer metastatic niche. Copyright © 2015 The Authors. Published by Elsevier Inc. All rights reserved.

Excess androgen during puberty disrupts circadian organization in female rats.

PubMed

Sellix, Michael T; Murphy, Zachary C; Menaker, Michael

2013-04-01

Circadian clocks have been described in each tissue of the hypothalamo-pituitary-ovarian axis. Although a role for the clock in the timing of ovulation is indicated, the impact of diseases that disrupt fertility on clock function or the clocks' role in the etiology of these pathologies has yet to be fully appreciated. Polycystic ovary syndrome (PCOS) is a particularly devastating endocrinopathy, affecting approximately 10% of women at childbearing age. Common features of PCOS are a polycystic ovary, amenorrhea, and excess serum androgen. Approximately 40% of these women have metabolic syndrome, including hyperinsulinemia, dyslipidemia, and hyperleptinemia. It has been suggested that excess androgen is a critical factor in the etiology of PCOS. We have examined the effects of androgen excess during puberty on the phase of circadian clocks in tissues of the metabolic and hypothalamo-pituitary-ovarian axes. Female period1-luciferase (per1-luc) rats were exposed to androgen (5α-dihydrotestosterone [DHT]) or placebo for 4-6 weeks (short term) or 9-15 weeks (long term). As expected, DHT-treated animals gained more weight than controls and had disrupted estrous cycles. At the end of treatment, tissues, including the liver, lung, kidney, white adipose, cornea, pituitary, oviduct, and ovarian follicles, were cultured, and per1-luc expression in each was recorded. Analysis of per1-luc expression revealed that DHT exposure increased phase distribution of multiple oscillators, including ovarian follicles, liver, and adipose, and altered phase synchrony between animals. These data suggest that excess androgen during puberty, a common feature of PCOS, negatively affects internal circadian organization in both the reproductive and metabolic axes.

Excess Androgen During Puberty Disrupts Circadian Organization in Female Rats

PubMed Central

Murphy, Zachary C.; Menaker, Michael

2013-01-01

Circadian clocks have been described in each tissue of the hypothalamo-pituitary-ovarian axis. Although a role for the clock in the timing of ovulation is indicated, the impact of diseases that disrupt fertility on clock function or the clocks' role in the etiology of these pathologies has yet to be fully appreciated. Polycystic ovary syndrome (PCOS) is a particularly devastating endocrinopathy, affecting approximately 10% of women at childbearing age. Common features of PCOS are a polycystic ovary, amenorrhea, and excess serum androgen. Approximately 40% of these women have metabolic syndrome, including hyperinsulinemia, dyslipidemia, and hyperleptinemia. It has been suggested that excess androgen is a critical factor in the etiology of PCOS. We have examined the effects of androgen excess during puberty on the phase of circadian clocks in tissues of the metabolic and hypothalamo-pituitary-ovarian axes. Female period1-luciferase (per1-luc) rats were exposed to androgen (5α-dihydrotestosterone [DHT]) or placebo for 4-6 weeks (short term) or 9-15 weeks (long term). As expected, DHT-treated animals gained more weight than controls and had disrupted estrous cycles. At the end of treatment, tissues, including the liver, lung, kidney, white adipose, cornea, pituitary, oviduct, and ovarian follicles, were cultured, and per1-luc expression in each was recorded. Analysis of per1-luc expression revealed that DHT exposure increased phase distribution of multiple oscillators, including ovarian follicles, liver, and adipose, and altered phase synchrony between animals. These data suggest that excess androgen during puberty, a common feature of PCOS, negatively affects internal circadian organization in both the reproductive and metabolic axes. PMID:23417420

The influences of a novel anti-adhesion device, thermally cross-linked gelatin film on peritoneal dissemination of tumor cells: The in vitro and in vivo experiments using murine carcinomatous peritonitis models.

PubMed

Miyamoto, Hiroe; Tsujimoto, Hiroyuki; Horii, Tsunehito; Ozamoto, Yuki; Ueda, Joe; Takagi, Toshitaka; Saitoh, Naoto; Hagiwara, Akeo

2017-10-10

To create anti-adhesive materials to be more effective and safer, we developed a thermally cross-linked gelatin film that showed superior anti-adhesive effects with excellent peritoneal regeneration. However, it may act as a convenient scaffold for tumor cell growth, thereby accelerating peritoneal dissemination when used in surgery for abdominal tumors. In this study, we tried to clarify this issue using mouse carcinomatous peritonitis models. First, we examined the in vitro tumor cell growth of mouse B16 melanoma or Colon26 cells on the gelatin film or the conventional hyarulonate/carboxymethylcellulose film. Tumor cell growth on each film was significantly lower than that of the control (no film). Next, we conducted the following in vivo experiments: After the parietal peritoneum was partially removed and covered with each film or without any film, mice were inoculated intraperitoneally with B16 melanoma or Colon26/Nluc cells expressing NanoLuc luciferase gene. At 7 days after the operation, we measured the weight of B16 melanoma tumors or the NanoLuc activity of Colon26/Nluc cells using in vivo imaging at the injured sites. There were no significant differences in the weight of the tumors and the NanoLuc activity among the three groups. We also observed the survival time of mice receiving the same operation and treatments. There was no significant difference in the survival time among the three groups. These results suggest that the gelatin film will likely not accelerate peritoneal dissemination as a convenient scaffold for tumor cell growth when used in surgery for abdominal tumors. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2017. © 2017 Wiley Periodicals, Inc.

Titanium biomaterials with complex surfaces induced aberrant peripheral circadian rhythms in bone marrow mesenchymal stromal cells.

PubMed

Hassan, Nathaniel; McCarville, Kirstin; Morinaga, Kenzo; Mengatto, Cristiane M; Langfelder, Peter; Hokugo, Akishige; Tahara, Yu; Colwell, Christopher S; Nishimura, Ichiro

2017-01-01

Circadian rhythms maintain a high level of homeostasis through internal feed-forward and -backward regulation by core molecules. In this study, we report the highly unusual peripheral circadian rhythm of bone marrow mesenchymal stromal cells (BMSCs) induced by titanium-based biomaterials with complex surface modifications (Ti biomaterial) commonly used for dental and orthopedic implants. When cultured on Ti biomaterials, human BMSCs suppressed circadian PER1 expression patterns, while NPAS2 was uniquely upregulated. The Ti biomaterials, which reduced Per1 expression and upregulated Npas2, were further examined with BMSCs harvested from Per1::luc transgenic rats. Next, we addressed the regulatory relationship between Per1 and Npas2 using BMSCs from Npas2 knockout mice. The Npas2 knockout mutation did not rescue the Ti biomaterial-induced Per1 suppression and did not affect Per2, Per3, Bmal1 and Clock expression, suggesting that the Ti biomaterial-induced Npas2 overexpression was likely an independent phenomenon. Previously, vitamin D deficiency was reported to interfere with Ti biomaterial osseointegration. The present study demonstrated that vitamin D supplementation significantly increased Per1::luc expression in BMSCs, though the presence of Ti biomaterials only moderately affected the suppressed Per1::luc expression. Available in vivo microarray data from femurs exposed to Ti biomaterials in vitamin D-deficient rats were evaluated by weighted gene co-expression network analysis. A large co-expression network containing Npas2, Bmal1, and Vdr was observed to form with the Ti biomaterials, which was disintegrated by vitamin D deficiency. Thus, the aberrant BMSC peripheral circadian rhythm may be essential for the integration of Ti biomaterials into bone.

Melanopsin resets circadian rhythms in cells by inducing clock gene Period1

NASA Astrophysics Data System (ADS)

Yamashita, Shuhei; Uehara, Tomoe; Matsuo, Minako; Kikuchi, Yo; Numano, Rika

2014-02-01

The biochemical, physiological and behavioral processes are under the control of internal clocks with the period of approximately 24 hr, circadian rhythms. The expression of clock gene Period1 (Per1) oscillates autonomously in cells and is induced immediately after a light pulse. Per1 is an indispensable member of the central clock system to maintain the autonomous oscillator and synchronize environmental light cycle. Per1 expression could be detected by Per1∷luc and Per1∷GFP plasmid DNA in which firefly luciferase and Green Fluorescence Protein were rhythmically expressed under the control of the mouse Per1 promoter in order to monitor mammalian circadian rhythms. Membrane protein, MELANOPSIN is activated by blue light in the morning on the retina and lead to signals transduction to induce Per1 expression and to reset the phase of circadian rhythms. In this report Per1 induction was measured by reporter signal assay in Per1∷luc and Per1∷GFP fibroblast cell at the input process of circadian rhythms. To the result all process to reset the rhythms by Melanopsin is completed in single cell like in the retina projected to the central clock in the brain. Moreover, the phase of circadian rhythm in Per1∷luc cells is synchronized by photo-activated Melanopsin, because the definite peak of luciferase activity in one dish was found one day after light illumination. That is an available means that physiological circadian rhythms could be real-time monitor as calculable reporter (bioluminescent and fluorescent) chronological signal in both single and groups of cells.

Reporter enzyme inhibitor study to aid assembly of orthogonal reporter gene assays.

PubMed

Ho, Pei-i; Yue, Kimberley; Pandey, Pramod; Breault, Lyne; Harbinski, Fred; McBride, Aaron J; Webb, Brian; Narahari, Janaki; Karassina, Natasha; Wood, Keith V; Hill, Adam; Auld, Douglas S

2013-05-17

Reporter gene assays (RGAs) are commonly used to measure biological pathway modulation by small molecules. Understanding how such compounds interact with the reporter enzyme is critical to accurately interpret RGA results. To improve our understanding of reporter enzymes and to develop optimal RGA systems, we investigated eight reporter enzymes differing in brightness, emission spectrum, stability, and substrate requirements. These included common reporter enzymes such as firefly luciferase (Photinus pyralis), Renilla reniformis luciferase, and β-lactamase, as well as mutated forms of R. reniformis luciferase emitting either blue- or green-shifted luminescence, a red-light emitting form of Luciola cruciata firefly luciferase, a mutated form of Gaussia princeps luciferase, and a proprietary luciferase termed "NanoLuc" derived from the luminescent sea shrimp Oplophorus gracilirostris. To determine hit rates and structure-activity relationships, we screened a collection of 42,460 PubChem compounds at 10 μM using purified enzyme preparations. We then compared hit rates and chemotypes of actives for each enzyme. The hit rates ranged from <0.1% for β-lactamase to as high as 10% for mutated forms of Renilla luciferase. Related luciferases such as Renilla luciferase mutants showed high degrees of inhibitor overlap (40-70%), while unrelated luciferases such as firefly luciferases, Gaussia luciferase, and NanoLuc showed <10% overlap. Examination of representative inhibitors in cell-based assays revealed that inhibitor-based enzyme stabilization can lead to increases in bioluminescent signal for firefly luciferase, Renilla luciferase, and NanoLuc, with shorter half-life reporters showing increased activation responses. From this study we suggest strategies to improve the construction and interpretation of assays employing these reporter enzymes.

Comparison of optical and power Doppler ultrasound imaging for non-invasive evaluation of arsenic trioxide as a vascular disrupting agent in tumors.

PubMed

Alhasan, Mustafa K; Liu, Li; Lewis, Matthew A; Magnusson, Jennifer; Mason, Ralph P

2012-01-01

Small animal imaging provides diverse methods for evaluating tumor growth and acute response to therapy. This study compared the utility of non-invasive optical and ultrasound imaging to monitor growth of three diverse human tumor xenografts (brain U87-luc-mCherry, mammary MCF7-luc-mCherry, and prostate PC3-luc) growing in nude mice. Bioluminescence imaging (BLI), fluorescence imaging (FLI), and Power Doppler ultrasound (PD US) were then applied to examine acute vascular disruption following administration of arsenic trioxide (ATO).During initial tumor growth, strong correlations were found between manual caliper measured tumor volume and FLI intensity, BLI intensity following luciferin injection, and traditional B-mode US. Administration of ATO to established U87 tumors caused significant vascular shutdown within 2 hrs at all doses in the range 5 to 10 mg/kg in a dose dependant manner, as revealed by depressed bioluminescent light emission. At lower doses substantial recovery was seen within 4 hrs. At 8 mg/kg there was >85% reduction in tumor vascular perfusion, which remained depressed after 6 hrs, but showed some recovery after 24 hrs. Similar response was observed in MCF7 and PC3 tumors. Dynamic BLI and PD US each showed similar duration and percent reductions in tumor blood flow, but FLI showed no significant changes during the first 24 hrs.The results provide further evidence for comparable utility of optical and ultrasound imaging for monitoring tumor growth, More specifically, they confirm the utility of BLI and ultrasound imaging as facile assays of the vascular disruption in solid tumors based on ATO as a model agent.

Playful Dialogues of a Bilingual Child in Everyday Conversations: Foundations of Early Literacy

ERIC Educational Resources Information Center

Soto Huerta, Mary Esther; Riojas-Cortez, Mari

2014-01-01

Many young children of Mexican heritage enter U.S. schools with knowledge of two language systems and emergent biliterate abilities. Bilingualism in young children may go unnoticed when instructional practices favor English. This case study describes how Lucía's bilingualism and emergent biliteracy parallel competencies essential to literacy…

The Implementation of a Geospatial Information Technology (GIT)-Supported Land Use Change Curriculum with Urban Middle School Learners to Promote Spatial Thinking

ERIC Educational Resources Information Center

Bodzin, Alec M.

2011-01-01

This study investigated whether a geospatial information technology (GIT)-supported science curriculum helped students in an urban middle school understand land use change (LUC) concepts and enhanced their spatial thinking. Five 8th grade earth and space science classes in an urban middle school consisting of three different ability level tracks…

Recombinant Platform for Prioritizing Aerolysin Molecular Grenades for Metastatic Prostate Cancer

DTIC Science & Technology

2015-10-01

hydrolysis occurring between glycine and phenylalanine (2)); Fibroblast activated protein (FAP) (i.e., ASGPAGPA] with hydrolysis occurring after the proline...PROBLEMS Nothing to Report ! 14! PRODUCTS 1. Recombinant Prodrug HSA/PA-PSA 2. C19-PSA LUC2 cell line ! 15! PARTICIPANTS & OTHER COLLABORATING ORGANIZATIONS Freddie L . Pruitt – NO CHANGE

The Denaturation of Environmental Education: Exploring the Role of Ecotechnologies

ERIC Educational Resources Information Center

Gough, Annette; Gough, Noel

2016-01-01

This article explores the changing ways "environment" has been represented in the discourses of environmental education and education for sustainable development (ESD) in United Nations (and related) publications since the 1970s. It draws on the writings of Jean-Luc Nancy and discusses the increasingly dominant view of the environment as…

Romanticism, Sexuality, and the Canon.

ERIC Educational Resources Information Center

Rowe, Kathleen K.

1990-01-01

Traces the Romanticism in the work and persona of film director Jean-Luc Godard. Examines the contradictions posed by Godard's politics and representations of sexuality. Asserts, that by bringing an ironic distance to the works of such canonized directors, viewers can take pleasure in those works despite their contradictions. (MM)

Light on curved backgrounds

NASA Astrophysics Data System (ADS)

Batic, D.; Nelson, S.; Nowakowski, M.

2015-05-01

We consider the motion of light on different spacetime manifolds by calculating the deflection angle, lensing properties and by probing into the possibility of bound states. The metrics in which we examine the light motion include, among other items, a general relativistic dark matter metric, a dirty black hole, and a worm hole metric, the last two inspired by noncommutative geometry. The lensing in a holographic screen metric is discussed in detail. We study also the bending of light around naked singularities like, e.g., the Janis-Newman-Winicour metric and include other cases. A generic property of light behavior in these exotic metrics is pointed out. For the standard metric like the Schwarzschild and Schwarzschild-de Sitter cases, we improve the accuracy of the lensing results for the weak and strong regimes.

PREFACE: Annual Conference on Functional Materials and Nanotechnologies - FM&NT 2011

NASA Astrophysics Data System (ADS)

Sternberg, Andris; Muzikante, Inta; Zicans, Janis

2011-06-01

The International Conference Functional Materials and Nanotechnologies (FM&NT-2011) was held in Riga, 5-8 April 2011 in the Institute of Solid State Physics, University of Latvia (ISSP LU). The conference was organized in co-operation with projects ERANET 'MATERA' and National Research programme in Materials Science and Information Technologies. The purpose of the conference was to bring together scientists, engineers and students from universities, research institutes and related industrial companies active in the field of advanced material science and materials technologies trends and future activities. Scientific themes covered in the conference are: theoretical research and modelling of processes and materials; materials for energetics, renewable energy technologies and phtovoltaics; multifunctional inorganic, organic and hybrid materials for photonic, micro and nanoelectronic applications and innovative methods for research of nanostructures; advanced technologies for synthesis and research of nanostructured materials, nanoparticles, thin films and coatings; application of innovative materials in science and economics. The number of registered participants from 17 countries was nearly 300. During three days of the conference 22 invited, 69 oral reports and 163 posters were presented. 40 papers, based on these reports, are included in this volume of IOP Conference Series: Materials Science and Engineering. Additional information about FM&NT-2011 is available in its homepage http://www.fmnt.lu.lv. The Organizing Committee would like to thank all speakers, contributors, session chairs, referees and meeting staff for their efforts in making the FM&NT-2011 successful. The Organizing Committee sincerely hopes that that the conference gave all participants new insights into the widespread development of functional materials and nanotechnologies and would enhance the circulation of information released at the meeting. Andris Sternberg Inta Muzikante Janis Zicans

The Role of Army Railroading at the Operational Level of War

DTIC Science & Technology

1989-05-15

Denmar China France Indonesia German,, Iran Greece Iraq Hungary Israel Italy Japan Latvia V< orea Li thuani a Lebanon Lu>c emo our g Saudi Arabia...Battalion -.. Pecan Street Jacksonville, Florida 7-2211-8719 Captain Horam Simpk ins U.S. Army Logistics Center ATTN: ATCL-E Fort Lee, Virginia 28:1

A Numerical Approach to Solving the Hall MHD Equations Including Diamagnetic Drift (Preprint)

DTIC Science & Technology

2008-02-19

SECURITY CLASSIFICATION OF: 17. LIMITATION OF ABSTRACT 18. NUMBER OF PAGES 19a. NAME OF RESPONSIBLE PERSON Dr. Jean-Luc Cambier a. REPORT...1997. [3] L. Chacon and D.A. Knoll. A 2d high-beta hall mhd implicit nonlinear solver. Journal of Computational Physics, 188:573–592, 2003. [4] Tony F

Regorafenib inhibits tumor progression through suppression of ERK/NF-κB activation in hepatocellular carcinoma bearing mice.

PubMed

Weng, Mao-Chi; Wang, Mei-Hui; Tsai, Jai-Jen; Kuo, Yu-Cheng; Liu, Yu-Chang; Hsu, Fei-Ting; Wang, Hsin-Ell

2018-06-29

Regorafenib has been demonstrated in our previous study to trigger apoptosis through suppression of extracellular signal-regulated kinase (ERK)/nuclear factor-κB (NF-κB) activation in hepatocellular carcinoma (HCC) SK-Hep1 cells in vitro However, the effect of regorafenib on NF-κB-modulated tumor progression in HCC in vivo is ambiguous. The aim of the present study is to investigate the effect of regorafenib on NF-κB-modulated tumor progression in HCC bearing mouse model. pGL4.50 luciferase reporter vector transfected SK-Hep1 (SK-Hep1/ luc2 ) and Hep3B 2.1-7 tumor bearing mice were established and used for the present study. Mice were treated with vehicle or regorafenib (20 mg/kg/day by gavage) for 14 days. Effects of regorafenib on tumor growth and protein expression together with toxicity of regorafenib were evaluated with digital caliper and bioluminescence imaging (BLI), ex vivo Western blotting immunohistochemistry (IHC) staining, and measurement of body weight and pathological examination of liver tissue, respectively, in SK-Hep1/ luc2 and Hep3B 2.1-7 tumor bearing mice. The results indicated regorafenib significantly reduced tumor growth and expression of phosphorylated ERK, NF-κB p65 (Ser536), phosphorylated AKT, and tumor progression-associated proteins. In addition, we found regorafenib induced both extrinsic and intrinsic apoptotic pathways. Body weight and liver morphology were not affected by regorafenib treatment. Our findings present the mechanism of tumor progression inhibition by regorafenib is linked to suppression of ERK/NF-κB signaling in SK-Hep1/ luc2 and Hep3B 2.1-7 tumor bearing mice. © 2018 The Author(s).

Intracellular cleavable poly(2-dimethylaminoethyl methacrylate) functionalized mesoporous silica nanoparticles for efficient siRNA delivery in vitro and in vivo.

PubMed

Lin, Daoshu; Cheng, Qiang; Jiang, Qian; Huang, Yuanyu; Yang, Zheng; Han, Shangcong; Zhao, Yuning; Guo, Shutao; Liang, Zicai; Dong, Anjie

2013-05-21

A low cytotoxicity and high efficiency delivery system with the advantages of low cost and facile fabrication is needed for the application of small interfering RNA (siRNA) delivery both in vitro and in vivo. For these prerequisites, cationic polymer-mesoporous silica nanoparticles (ssCP-MSNs) were prepared by surface functionalized mesoporous silica nanoparticles with disulfide bond cross-linked poly(2-dimethylaminoethyl methacrylate) (PDMAEMA). In vitro and in vivo evaluations were performed. The synthesized ssCP-MSNs are 100-150 nm in diameter with a pore size of 10 nm and a positively charged surface with a high zeta potential of 27 mV. Consequently, the ssCP-MSNs showed an excellent binding capacity for siRNA, and an enhancement in the cell uptake and cytosolic availability of siRNA. Furthermore, the intracellular reducing cleavage of the disulfide bonds cross-linking the PDMAEMA segments led to intracellular cleavage of PDMAEMA from ssCP-MSNs, which facilitated the intracellular triggered release of siRNA. Therefore, promoted RNA interference was observed in HeLa-Luc cells, which was equal to that of Lipofectamine 2000. Significantly, compared to Lipofectamine 2000, the ssCP-MSNs were more biocompatible, with low cytotoxicity (even non-cytotoxicity) and promotion of cell proliferation to HeLa-Luc cells. The in vivo systemic distribution studies certified that ssCP-MSNs/siRNA could prolong the duration of siRNA in vivo, and that they accumulated in the adrenal gland, liver, lung, spleen, kidney, heart and thymus after intravenous injection. Encouragingly, with the ability to deliver siRNA to a tumor, ssCP-MSNs/siRNA showed a tumor suppression effect in the HeLa-Luc xenograft murine model after intravenous injection. Therefore, the ssCP-MSNs cationic polymer-mesoporous silica nanoparticles with low cytotoxicity are promising for siRNA delivery.

Infectivity of attenuated poxvirus vaccine vectors and immunogenicity of a raccoonpox vectored rabies vaccine in the Brazilian Free-tailed bat (Tadarida brasiliensis)

USGS Publications Warehouse

Stading, Benjamin; Osorio, Jorge E.; Velasco-Villa, Andres; Smotherman, Michael; Kingstad-Bakke, Brock; Rocke, Tonie E.

2016-01-01

Bats (Order Chiroptera) are an abundant group of mammals with tremendous ecological value as insectivores and plant dispersers, but their role as reservoirs of zoonotic diseases has received more attention in the last decade. With the goal of managing disease in free-ranging bats, we tested modified vaccinia Ankara (MVA) and raccoon poxvirus (RCN) as potential vaccine vectors in the Brazilian Free-tailed bat (Tadarida brasiliensis), using biophotonic in vivo imaging and immunogenicity studies. Animals were administered recombinant poxviral vectors expressing the luciferase gene (MVA-luc, RCN-luc) through oronasal (ON) or intramuscular (IM) routes and subsequently monitored for bioluminescent signal indicative of viral infection. No clinical illness was noted after exposure to any of the vectors, and limited luciferase expression was observed. Higher and longer levels of expression were observed with the RCN-luc construct. When given IM, luciferase expression was limited to the site of injection, while ON exposure led to initial expression in the oral cavity, often followed by secondary replication at another location, likely the gastric mucosa or gastric associated lymphatic tissue. Viral DNA was detected in oral swabs up to 7 and 9 days post infection (dpi) for MVA and RCN, respectively. While no live virus was detected in oral swabs from MVA-infected bats, titers up to 3.88 x 104 PFU/ml were recovered from oral swabs of RCN-infected bats. Viral DNA was also detected in fecal samples from two bats inoculated IM with RCN, but no live virus was recovered. Finally, we examined the immunogenicity of a RCN based rabies vaccine (RCN-G) following ON administration. Significant rabies neutralizing antibody titers were detected in the serum of immunized bats using the rapid fluorescence focus inhibition test (RFFIT). These studies highlight the safety and immunogenicity of attenuated poxviruses and their potential use as vaccine vectors in bats.

Infectivity of attenuated poxvirus vaccine vectors and immunogenicity of a raccoonpox vectored rabies vaccine in the Brazilian Free-tailed bat (Tadarida brasiliensis)

PubMed Central

Stading, Ben R.; Osorio, Jorge E.; Velasco-Villa, Andres; Smotherman, Michael; Kingstad-Bakke, Brock

2017-01-01

Bats (Order Chiroptera) are an abundant group of mammals with tremendous ecological value as insectivores and plant dispersers, but their role as reservoirs of zoonotic diseases has received more attention in the last decade. With the goal of managing disease in free-ranging bats, we tested modified vaccinia Ankara (MVA) and raccoon poxvirus (RCN) as potential vaccine vectors in the Brazilian Free-tailed bat (Tadarida brasiliensis), using biophotonic in vivo imaging and immunogenicity studies. Animals were administered recombinant poxviral vectors expressing the luciferase gene (MVA-luc, RCN-luc) through oronasal (ON) or intramuscular (IM) routes and subsequently monitored for bioluminescent signal indicative of viral infection. No clinical illness was noted after exposure to any of the vectors, and limited luciferase expression was observed. Higher and longer levels of expression were observed with the RCN-luc construct. When given IM, luciferase expression was limited to the site of injection, while ON exposure led to initial expression in the oral cavity, often followed by secondary replication at another location, likely the gastric mucosa or gastric associated lymphatic tissue. Viral DNA was detected in oral swabs up to 7 and 9 days post infection (dpi) for MVA and RCN, respectively. While no live virus was detected in oral swabs from MVA-infected bats, titers up to 3.88 × 104 PFU/ml were recovered from oral swabs of RCN-infected bats. Viral DNA was also detected in fecal samples from two bats inoculated IM with RCN, but no live virus was recovered. Finally, we examined the immunogenicity of a RCN based rabies vaccine (RCN-G) following ON administration. Significant rabies neutralizing antibody titers were detected in the serum of immunized bats using the rapid fluorescence focus inhibition test (RFFIT). These studies highlight the safety and immunogenicity of attenuated poxviruses and their potential use as vaccine vectors in bats. PMID:27650872

Systemic Immunotherapy of Non-Muscle Invasive Mouse Bladder Cancer with Avelumab, an Anti-PD-L1 Immune Checkpoint Inhibitor.

PubMed

Vandeveer, Amanda J; Fallon, Jonathan K; Tighe, Robert; Sabzevari, Helen; Schlom, Jeffrey; Greiner, John W

2016-05-01

Bacillus Calmette-Guerin (BCG) is the standard of care for intravesical therapy for carcinoma in situ and non-muscle invasive, nonmetastatic human urothelial carcinoma. Although the responsiveness to this immunotherapeutic is believed to be linked with (i) a high number of somatic mutations and (ii) a large number of tumor-infiltrating lymphocytes, recent findings of the roles that inhibitory immune receptors and their ligands play in tumor evasion may provide insights into the limitations of the effectiveness of BCG and offer new targets for immune-based therapy. In this study, an aggressive, bioluminescent orthotopic bladder cancer model, MB49 tumor cells transfected with luciferase (MB49(luc)), was used to study the antitumor effects of avelumab, an antibody to PD-L1. MB49(luc) murine tumor cells form multifocal tumors on the mucosal wall of the bladder reminiscent of non-muscle invasive, nonmetastatic urothelial carcinomas. MB49(luc) bladder tumors are highly positive for the expression of PD-L1, and avelumab administration induced significant (P < 0.05) antitumor effects. These antitumor effects were more dependent on the presence of CD4 than CD8 T cells, as determined by in vivo immune cell depletions. The findings suggest that in this bladder tumor model, interruption of the immune-suppressive PD-1/PD-L1 complex releases a local adaptive immune response that, in turn, reduces tumor growth. This bladder tumor model can be used to further identify host antitumor immune mechanisms and evaluate combinations of immune-based therapies for carcinoma in situ and non-muscle invasive, nonmetastatic urothelial carcinoma, to provide the rationale for subsequent clinical studies. Cancer Immunol Res; 4(5); 452-62. ©2016 AACR. ©2016 American Association for Cancer Research.

Prediction of future urban growth using CA-Markov for urban sustainability planning of Banda Aceh, Indonesia

NASA Astrophysics Data System (ADS)

Achmad, A.; Irwansyah, M.; Ramli, I.

2018-03-01

Banda Aceh experienced rapid growth, both physically, socially, and economically, after the Tsunami that devastated it the end of December in 2004. Hence policy controls are needed to direct the pattern of urban growth to achieve sustainable development for the future. The purpose of this paper is to generate a growth model for Banda Aceh using the CA-Markov process. By knowing the changes in land use between 2005 and 2009 from the results of previous research, simulations for 2013, 2019 and 2029 using the application of Idrisi@Selva. CA-Markov models were prepared to determine the quantity of changes. The simulation results showed that, after the Tsunami, the City of Banda Aceh tended to grow towards the coast. For the control of the LUC, the Banda Aceh City government needs to prepare comprehensive and detailed maps and inventory of LUC for the city to provide basic data and information needed for monitoring and evaluation that can be done effectively and efficiently. An institution for monitoring and evaluation of the urban landscape and the LUC should be formed immediately. This institution could consist of representatives from government, academia, community leaders, the private sector and other experts. The findings from this study can be used to start the monitoring and evaluation of future urban growth. Especially for the coastal areas, the local government should immediately prepare special spatial coastal area plans to control growth in those areas and to ensure that the economic benefits from disaster mitigation and coastal protection are preserved. For the development of the city in the future, it is necessary to achieve a balance between economic development, and social welfare with environmental protection and disaster mitigation. iIt will become a big challenge to achieve sustainable development for the future.

Sensitization of B16 tumor cells with a CXCR4 antagonist increases the efficacy of immunotherapy for established lung metastases

PubMed Central

Lee, Chih-hung; Kakinuma, Takashi; Wang, Julia; Zhang, Hong; Palmer, Douglas C.; Restifo, Nicholas P.; Hwang, Sam T.

2008-01-01

Expression of the chemokine receptor CXCR4 by tumor cells promotes metastasis, possibly by activating pro-survival signals that render cancer cells resistant to immune attack. Inhibition of CXCR4 with a peptide antagonist, T22, blocks metastatic implantation of CXCR4-transduced B16 (CXCR4-luc-B16) melanoma cells in lung, but not the outgrowth of established metastases, raising the question of how T22 can best be used in a clinical setting. Herein, whereas the treatment of CXCR4-luc-B16 cells in vitro with the CXCR4 ligand CXCL12 did not reduce killing induced by cisplatin or cyclophosphamide, CXCL12 markedly reduced Fas-dependent killing by gp100-specific (pmel-1) CD8+ T cells. T22 pretreatment restored sensitivity of CXCR4-luc-B16 cells to pmel-1 killing, even in the presence of CXCL12. Two immune-augmenting regimens were used in combination with T22 to treat experimental lung metastases. First, low-dose cyclophosphamide treatment (100 mg/kg) on day 5 in combination with T22 (days 4–7) yielded a ~70% reduction of B16 metastatic tumor burden in the lungs compared with cyclophosphamide treatment alone (P < 0.001). Furthermore, whereas anti–CTL antigen 4 (CTLA4) monoclonal antibody (mAb; or T22 treatment) alone had little effect on established B16 metastases, pretreatment with T22 (in combination with anti-CTLA4 mAb) resulted in a 50% reduction in lung tumor burden (P = 0.02). Thus, in vitro, CXCR4 antagonism with T22 renders B16 cells susceptible to killing by antigen-specific T cells. In vivo, T22 synergizes with cyclophosphamide or anti-CTLA4 mAb in the treatment of established lung metastases, suggesting a novel strategy for augmenting the efficacy of immunotherapy. PMID:17041104

Input-driven versus turnover-driven controls of simulated changes in soil carbon due to land-use change

NASA Astrophysics Data System (ADS)

Nyawira, Sylvia; Nabel, Julia; Brovkin, Victor; Pongratz, Julia

2017-04-01

Modelling studies estimate a global loss in soil carbon caused by land-use changes (LUCs) over the last century. Although it is known that this loss stems from the changes in quantity of litter inputs from the vegetation to the soil (input-driven) and the changes in turnover of carbon in the soil (turnover-driven) associated with LUC, the individual contribution of these two controls to the total changes have not been assessed. Using the dynamic global vegetation model JSBACH, we apply a factor separation approach to isolate the contribution of the input-driven and turnover-driven changes, as well as their synergies, to the total changes in soil carbon from LUC. To assess how land management through crop and wood harvest influences the controls, we compare our results for simulations with and without land management. Our results reveal that for the afforested regions both the input-driven and turnover-driven changes generally result in soil carbon gain, whereas deforested regions exhibit a loss. However, for regions where croplands have increased at the expense of grasslands and pastures, the input-driven changes result in a loss that is partly offset by a gain via the turnover-driven changes. This gain stems from a decrease in the fire-related carbon losses when grasslands or pastures are replaced with croplands. Omitting land management reduces the carbon losses in regions where natural vegetation has been converted to croplands and enhances the gain in afforested regions. The global simulated losses are substantially reduced from 54.0 Pg C to 22.0 Pg C, with the input-driven losses reducing from 54.7 Pg C to 24.9 Pg C. Our study shows that the dominating control of soil carbon losses is through the input-driven changes, which are more directly influenced by human management than the turnover-driven ones.

AAVPG: A vigilant vector where transgene expression is induced by p53

DOE Office of Scientific and Technical Information (OSTI.GOV)

Bajgelman, Marcio C.; Medrano, Ruan F.V.; Carvalho, Anna Carolina P.V.

2013-12-15

Using p53 to drive transgene expression from viral vectors may provide on demand expression in response to physiologic stress, such as hypoxia or DNA damage. Here we introduce AAVPG, an adeno-associated viral (AAV) vector where a p53-responsive promoter, termed PG, is used to control transgene expression. In vitro assays show that expression from the AAVPG-luc vector was induced specifically in the presence of functional p53 (1038±202 fold increase, p<0.001). The AAVPG-luc vector was an effective biosensor of p53 activation in response to hypoxia (4.48±0.6 fold increase in the presence of 250 µM CoCl{sub 2}, p<0.001) and biomechanical stress (2.53±0.4 foldmore » increase with stretching, p<0.05). In vivo, the vigilant nature of the AAVPG-luc vector was revealed after treatment of tumor-bearing mice with doxorubicin (pre-treatment, 3.4×10{sup 5}±0.43×10{sup 5} photons/s; post-treatment, 6.6×10{sup 5}±2.1×10{sup 5} photons/s, p<0.05). These results indicate that the AAVPG vector is an interesting option for detecting p53 activity both in vitro and in vivo. - Highlights: • AAV vector where transgene expression is controlled by the tumor suppressor p53. • The new vector, AAVPG, shown to function as a biosensor of p53 activity, in vitro and in vivo. • The p53 activity monitored by the AAVPG vector is relevant to cancer and other diseases. • AAVPG reporter gene expression was activated upon DNA damage, hypoxia and mechanical stress.« less

Landscape-based upstream-downstream prevalence of land-use/cover change drivers in southeastern rift escarpment of Ethiopia.

PubMed

Temesgen, Habtamu; Wu, Wei; Legesse, Abiyot; Yirsaw, Eshetu; Bekele, Belew

2018-02-23

Characterized by high population density on a rugged topography, the Gedeo-Abaya landscape dominantly contains a multi-strata traditional agroforests showing the insight of Gedeo farmers on natural resource management practices. Currently, this area has been losing its resilience and is becoming unable to sustain its inhabitants. Based on both RS-derived and GIS-computed land-use/cover changes (LUCC) as well as socioeconomic validations, this article explored the LUCC and agroecological-based driver patterns in Gedeo-Abaya landscape from 1986 to 2015. A combination of geo-spatial technology and cross-sectional survey design were employed to detect the drivers behind these changes. The article discussed that LUCC and the prevalence of drivers are highly diverse and vary throughout agroecological zones. Except for the population, most downstream top drivers are perceived as insignificant in the upstream region and vice versa. In the downstream, land-use/cover (LUC) classes are more dynamic, diverse, and challenged by nearly all anticipated drivers than are upstream ones. Agroforestry LUC has been increasing (by 25% of its initial cover) and is becoming the predominant cover type, although socioeconomic analysis and related findings show its rapid LUC modification. A rapid reduction of woodland/shrubland (63%) occurred in the downstream, while wetland/marshy land increased threefold (158%), from 1986 to 2015 with annual change rates of - 3.7 and + 6%, respectively. Land degradation induced by changes in land use is a serious problem in Africa, especially in the densely populated sub-Saharan regions such as Ethiopia (FAO 2015). Throughout the landscape, LUCC is prominently affecting land-use system of the study landscape due to population pressure in the upstream region and drought/rainfall variability, agribusiness investment, and charcoaling in the downstream that necessitate urgent action.

Optimisations and Challenges Involved in the Creation of Various Bioluminescent and Fluorescent Influenza A Virus Strains for In Vitro and In Vivo Applications

PubMed Central

Herfst, Sander; Bestebroer, Theo M.; Vaes, Vincent P.; van der Hoeven, Barbara; Koster, Abraham J.; Kremers, Gert-Jan; Scott, Dana P.; Gultyaev, Alexander P.; Sorell, Erin M.; de Graaf, Miranda; Bárcena, Montserrat; Rimmelzwaan, Guus F.; Fouchier, Ron A.

2015-01-01

Bioluminescent and fluorescent influenza A viruses offer new opportunities to study influenza virus replication, tropism and pathogenesis. To date, several influenza A reporter viruses have been described. These strategies typically focused on a single reporter gene (either bioluminescent or fluorescent) in a single virus backbone. However, whilst bioluminescence is suited to in vivo imaging, fluorescent viruses are more appropriate for microscopy. Therefore, the idea l reporter virus varies depending on the experiment in question, and it is important that any reporter virus strategy can be adapted accordingly. Herein, a strategy was developed to create five different reporter viruses in a single virus backbone. Specifically, enhanced green fluorescent protein (eGFP), far-red fluorescent protein (fRFP), near-infrared fluorescent protein (iRFP), Gaussia luciferase (gLUC) and firefly luciferase (fLUC) were inserted into the PA gene segment of A/PR/8/34 (H1N1). This study provides a comprehensive characterisation of the effects of different reporter genes on influenza virus replication and reporter activity. In vivo reporter gene expression, in lung tissues, was only detected for eGFP, fRFP and gLUC expressing viruses. In vitro, the eGFP-expressing virus displayed the best reporter stability and could be used for correlative light electron microscopy (CLEM). This strategy was then used to create eGFP-expressing viruses consisting entirely of pandemic H1N1, highly pathogenic avian influenza (HPAI) H5N1 and H7N9. The HPAI H5N1 eGFP-expressing virus infected mice and reporter gene expression was detected, in lung tissues, in vivo. Thus, this study provides new tools and insights for the creation of bioluminescent and fluorescent influenza A reporter viruses. PMID:26241861

"Detecting people"

NASA Astrophysics Data System (ADS)

Arneth, A.; Pugh, T.; Krause, A.; Bayer, A.; Lindeskog, M.

2015-12-01

Land-use change (LUC) is known to significantly affect biogeochemical cycles as well as surface energy partitioning - with important implications, ranging from understanding present-day measurements, to simulations of climate change and impacts on ecosystems, to assessments of the mitigation potential of land-based mitigation policies on ecosystems. When connecting observations of surface-atmosphere interactions and modelling at different scales, two important issues in this context are: legacy effects (e.g., to what degree and for how long does past LUC at a given location affect vegetation structure, CO2 fluxes and carbon pools), and sub-grid variability of the land-use change per se (e.g., whether bi-directional information about changes are taken into consideration). Both are important when bridging between scales (in time and in space) to enhance long-term observation networks. This contribution to the session will be very much from a process-based modelling perspective. Using a second generation dynamic global vegetation model we will show how different land-use histories impact vegetation and soil recovery (carbon pool-size, fluxes) differently, depending on the type of previous land-use, its length, and on the type of biome. We also study the difference between "gross" and "net" LUC accounting for simulated carbon cycling. Two important aspects, considering the session's objectives, are: 1) When establishing and developing observation networks, land-use history is key information for the interpretation of measured fluxes and needs to be collected and made available;, 2) Observation networks that "operate" solely in the natural science domain need to increasingly seek cooperation with socio-economic observations (such as land-use change, land management) in order to gain better understanding of coupled socio-ecological systems.

Suprachiasmatic nucleus function and circadian entrainment are modulated by G protein-coupled inwardly rectifying (GIRK) channels

PubMed Central

Hablitz, L M; Molzof, H E; Paul, J R; Johnson, R L; Gamble, K L

2014-01-01

Abstract G protein signalling within the central circadian oscillator, the suprachiasmatic nucleus (SCN), is essential for conveying time-of-day information. We sought to determine whether G protein-coupled inwardly rectifying potassium channels (GIRKs) modulate SCN physiology and circadian behaviour. We show that GIRK current and GIRK2 protein expression are greater during the day. Pharmacological inhibition of GIRKs and genetic loss of GIRK2 depolarized the day-time resting membrane potential of SCN neurons compared to controls. Behaviourally, GIRK2 knockout (KO) mice failed to shorten free running period in response to wheel access in constant darkness and entrained more rapidly to a 6 h advance of a 12 h:12 h light–dark (LD) cycle than wild-type (WT) littermate controls. We next examined whether these effects were due to disrupted signalling of neuropeptide Y (NPY), which is known to mediate non-photic phase shifts, attenuate photic phase shifts and activate GIRKs. Indeed, GIRK2 KO SCN slices had significantly fewer silent cells in response to NPY, likely contributing to the absence of NPY-induced phase advances of PER2::LUC rhythms in organotypic SCN cultures from GIRK2 KO mice. Finally, GIRK channel activation is sufficient to cause a non-photic-like phase advance of PER2::LUC rhythms on a Per2Luc+/− background. These results suggest that rhythmic regulation of GIRK2 protein and channel function in the SCN contributes to day-time resting membrane potential, providing a mechanism for the fine tuning responses to non-photic and photic stimuli. Further investigation could provide insight into disorders with circadian disruption comorbidities such as epilepsy and addiction, in which GIRK channels have been implicated. PMID:25217379

Molecular and functional evaluation of a novel HIF inhibitor, benzopyranyl 1,2,3-triazole compound

PubMed Central

Park, Kyunghye; Lee, Hye Eun; Lee, Sun Hee; Lee, Doohyun; Lee, Taeho; Lee, You Mie

2017-01-01

Hypoxia occurs in a variety of pathological events, including the formation of solid tumors. Hypoxia-inducible factor (HIF)-1α is stabilized under hypoxic conditions and is a key molecule in tumor growth and angiogenesis. Seeking to develop novel cancer therapeutics, we investigated small molecules from our in-house chemical libraries to target HIF-1α. We employed a dual-luciferase assay that uses a luciferase (Luc) reporter vector harboring five copies of hypoxia-responsive element (HRE) in the promoter. Under hypoxic conditions that increased Luc reporter activity by four-fold, we screened 144 different compounds, nine of which showed 30–50% inhibition of hypoxia-induced Luc reporter activity. Among these, “Compound 12, a benzopyranyl 1,2,3-triazole” was the most efficient at inhibiting the expression of HIF-1α under hypoxic conditions, reducing its expression by 80%. Under hypoxic conditions, the half maximal IC50 of the compound was 24 nM in HEK-293 human embryonic kidney cells, and 2 nM in A549 human lung carcinoma cells. Under hypoxic conditions, Compound 12 increased hydroxylated HIF-1α levels and HIF-1α ubiquitination, and also dose-dependently decreased HIF-1α target gene expression as well as vascular endothelial growth factor (VEGF) secretion. Furthermore, this compound inhibited VEGF-induced in vitro angiogenesis in human umbilical vein endothelial cells (HUVECs), and in vivo, it inhibited chick chorioallantoic membrane angiogenesis. In allogaft assays, cotreatment with Compound 12 and gefitinib significantly inhibited tumor growth and angiogenesis. Compound 12 can be a novel inhibitor of HIF-1α by accelerating its degradation, and shows much potential as an anti-cancer agent through its ability to suppress tumor growth and angiogenesis. PMID:27999195

Developmental programming by androgen affects the circadian timing system in female mice.

PubMed

Mereness, Amanda L; Murphy, Zachary C; Sellix, Michael T

2015-04-01

Circadian clocks play essential roles in the timing of events in the mammalian hypothalamo-pituitary-ovarian (HPO) axis. The molecular oscillator driving these rhythms has been localized to tissues of the HPO axis. It has been suggested that synchrony among these oscillators is a feature of normal reproductive function. The impact of fertility disorders on clock function and the role of the clock in the etiology of endocrine pathology remain unknown. Polycystic ovarian syndrome (PCOS) is a particularly devastating fertility disorder, affecting 5%-10% of women at childbearing age with features including a polycystic ovary, anovulation, and elevated serum androgen. Approximately 40% of these women have metabolic syndrome, marked by hyperinsulinemia, dyslipidemia, and insulin resistance. It has been suggested that developmental exposure to excess androgen contributes to the etiology of fertility disorders, including PCOS. To better define the role of the timing system in these disorders, we determined the effects of androgen-dependent developmental programming on clock gene expression in tissues of the metabolic and HPO axes. Female PERIOD2::luciferase (PER2::LUC) mice were exposed to androgen (dihydrotestosterone [DHT]) in utero (Days 16-18 of gestation) or for 9-10 wk (DHT pellet) beginning at weaning (pubertal androgen excess [PAE]). As expected, both groups of androgen-treated mice had disrupted estrous cycles. Analysis of PER2::LUC expression in tissue explants revealed that excess androgen produced circadian misalignment via tissue-dependent effects on phase distribution. In vitro treatment with DHT differentially affected the period of PER2::LUC expression in tissue explants and granulosa cells, indicating that androgen has direct and tissue-specific effects on clock gene expression that may account for the effects of developmental programming on the timing system. © 2015 by the Society for the Study of Reproduction, Inc.

Characterization and distribution of trace organic contaminants in sediment from Masan Bay, Korea. 2: In vitro gene expression assays

DOE Office of Scientific and Technical Information (OSTI.GOV)

Khim, J.S.; Villeneuve, D.L.; Kannan, K.

1999-12-01

Extracts of sediment collected from Masan Bay, Korea were fractionated using Florisil columns, Fractions were screened for their ability to induce estrogen- and dioxini-like gene expression in vitro using NVLN and H2IIE-luc cells, respectively, both before and after acid treatment. Florisil fraction 1 (F1), which was shown to contain polychlorinated biphenyls, induced very little response in either assay. The midpolarity Florisil fraction (F2) was the most active fraction. Twenty-seven of 28 F2 samples induced significant estrogenic activity, and all 28 samples induced significant dioxin-like activity. Twelve of the F2 samples produced magnitudes of response in the dioxin-responsive H2IIIE-luc cells similarmore » to those induced by a 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) standard, Acid treatment did not markedly diminish the activity of F2 samples. These results suggested the presence of unidentified, acid stable, aryl hydrocarbon receptor (AhR) and estrogen receptor (ER) agonists in F2. Twenty-three of the 28 most polar florisil fractions (F3) were either cytotoxic or caused morphological changes in estrogen-responsive NVLN cells, while over half of the F3 samples caused similar effects in H2IIE-luc cells. Empirical evidence suggested that acid labile compounds contributed to both the estrogenic and cytotoxic responses of the NVLN cells. Mass balance suggested that known concentrations of alkylphenols and bisphenol A may account for a portion of the estrogenic response but were not great enough to account for the toxicity. Acid labile compounds also contributed substantially to the dioxin-like activity of F3 samples. This adds to a growing body of evidence which suggests the presence of unidentified, relatively polar, aryl hydrocarbon receptor agonists in sediment from some areas.« less

Soil carbon sequestration and land use change associated with biofuel production: Empirical evidence

DOE Office of Scientific and Technical Information (OSTI.GOV)

Qin, Zhangcai; Dunn, Jennifer B.; Kwon, Hoyoung

Soil organic carbon (SOC) change can be a major impact of land use change (LUC) associated with biofuel feedstock production. By collecting and analyzing data from worldwide field observations with major LUCs from cropland, grassland and forest to lands producing biofuel crops (i.e., corn, switchgrass, Miscanthus, poplar and willow), we were able to estimate SOC response ratios and sequestration rates and evaluate the effects of soil depth and time scale on SOC change. Both the amount and rate of SOC change were highly dependent on the specific land transition. Irrespective of soil depth or time horizon, cropland conversions resulted inmore » an overall SOC gain of 6-14% relative to initial SOC level, while conversion from grassland or forest to corn (without residue removal) or poplar caused significant carbon loss (9-35%). No significant SOC changes were observed in land converted from grasslands or forests to switchgrass, Miscanthus or willow. The SOC response ratios were similar in both 0-30 and 0-100 cm soil depths in most cases, suggesting SOC changes in deep soil and that use of top soil only for SOC accounting in biofuel life cycle analysis (LCA) might underestimate total SOC changes. Soil carbon sequestration rates varied greatly among studies and land transition types. Generally, the rates of SOC change tended to be the greatest during the 10 years following land conversion, and had declined to approach 0 within about 20 years for most LUCs. Observed trends in SOC change were generally consistent with previous reports. Soil depth and duration of study significantly influence SOC change rates and so should be considered in carbon emission accounting in biofuel LCA. High uncertainty remains for many perennial systems, field trials and modeling efforts are needed to determine the site- and system-specific rates and direction of change associated with their production.« less

Augmentation of the posterior maxilla: a proposed hierarchy of treatment selection.

PubMed

Fugazzotto, Paul A

2003-11-01

Literature is reviewed that discusses treatment results following Cauldwel Luc approach sinus augmentation therapy or osteotome sinus augmentation therapy, with and without simultaneous implant placement. A hierarchy of treatment selection for the augmentation of the posterior maxilla, based upon quantity and position of residual alveolar bone crestal to the floor of the sinus, is proposed.

Challenges Facing Higher Education at the Millennium. American Council on Education/Oryx Press Series on Higher Education.

ERIC Educational Resources Information Center

Hirsch, Werner Z., Ed.; Weber, Luc E., Ed.

This collection of 17 essays focuses on the major changes that higher education needs to make so that it can effectively pursue teaching and research and significantly contribute public service in a rapidly changing world. The essays include: (1) "Survey of the Main Challenges Facing Higher Education at the Millennium" (Luc E. Weber);…

76 FR 28781 - Independent Scientific Peer Review Panel Report: Evaluation of the Validation Status of an In...

Federal Register 2010, 2011, 2012, 2013, 2014

2011-05-18

... 29-30, 2011, to evaluate the validation status of the LUMI-CELL[supreg] (BG1Luc ER TA) test method...: http://iccvam.niehs.nih.gov/docs/endo_docs/EDPRPRept2011.pdf or by contacting NICEATM (see ADDRESSES). The report contains (1) the Panel's evaluation of the validation status of the test method and (2) the...

Godard on Godard.

ERIC Educational Resources Information Center

Narboni, Jean, Ed.; Milne, Tom, Ed.

The critical writings of the French film director, Jean-Luc Godard, are collected here. Many of them are reviews which originally appeared in the French magazines "Cahiers du Cinema" and "La Gazette du Cinema." They include comments on many American and European films from the years 1950-59, as well as Godard's comments on…

Nucleic acid detection using BRET-beacons based on bioluminescent protein-DNA hybrids.

PubMed

Engelen, Wouter; van de Wiel, Kayleigh M; Meijer, Lenny H H; Saha, Bedabrata; Merkx, Maarten

2017-03-02

Bioluminescent molecular beacons have been developed using a modular design approach that relies on BRET between the bright luciferase NanoLuc and a Cy3 acceptor. While classical molecular beacons are hampered by background fluorescence and scattering, these BRET-beacons allow detection of low pM concentrations of nucleic acids directly in complex media.

A bright cyan-excitable orange fluorescent protein facilitates dual-emission microscopy and enhances bioluminescence imaging in vivo

PubMed Central

Chu, Jun; Oh, Young-Hee; Sens, Alex; Ataie, Niloufar; Dana, Hod; Macklin, John J.; Laviv, Tal; Welf, Erik S.; Dean, Kevin M.; Zhang, Feijie; Kim, Benjamin B.; Tang, Clement Tran; Hu, Michelle; Baird, Michelle A.; Davidson, Michael W.; Kay, Mark A.; Fiolka, Reto; Yasuda, Ryohei; Kim, Douglas S.; Ng, Ho-Leung; Lin, Michael Z.

2016-01-01

Orange-red fluorescent proteins (FPs) are widely used in biomedical research for multiplexed epifluorescence microscopy with GFP-based probes, but their different excitation requirements make multiplexing with new advanced microscopy methods difficult. Separately, orange-red FPs are useful for deep-tissue imaging in mammals due to the relative tissue transmissibility of orange-red light, but their dependence on illumination limits their sensitivity as reporters in deep tissues. Here we describe CyOFP1, a bright engineered orange-red FP that is excitable by cyan light. We show that CyOFP1 enables single-excitation multiplexed imaging with GFP-based probes in single-photon and two-photon microscopy, including time-lapse imaging in light-sheet systems. CyOFP1 also serves as an efficient acceptor for resonance energy transfer from the highly catalytic blue-emitting luciferase NanoLuc. An optimized fusion of CyOFP1 and NanoLuc, called Antares, functions as a highly sensitive bioluminescent reporter in vivo, producing substantially brighter signals from deep tissues than firefly luciferase and other bioluminescent proteins. PMID:27240196

Comparing environmental consequences of anaerobic mono- and co-digestion of pig manure to produce bio-energy--a life cycle perspective.

PubMed

De Vries, J W; Vinken, T M W J; Hamelin, L; De Boer, I J M

2012-12-01

The aim of this work was to assess the environmental consequences of anaerobic mono- and co-digestion of pig manure to produce bio-energy, from a life cycle perspective. This included assessing environmental impacts and land use change emissions (LUC) required to replace used co-substrates for anaerobic digestion. Environmental impact categories considered were climate change, terrestrial acidification, marine and freshwater eutrophication, particulate matter formation, land use, and fossil fuel depletion. Six scenarios were evaluated: mono-digestion of manure, co-digestion with: maize silage, maize silage and glycerin, beet tails, wheat yeast concentrate (WYC), and roadside grass. Mono-digestion reduced most impacts, but represented a limited source for bio-energy. Co-digestion with maize silage, beet tails, and WYC (competing with animal feed), and glycerin increased bio-energy production (up to 568%), but at expense of increasing climate change (through LUC), marine eutrophication, and land use. Co-digestion with wastes or residues like roadside grass gave the best environmental performance. Copyright © 2012 Elsevier Ltd. All rights reserved.

Selection of mRNA 5'-untranslated region sequence with high translation efficiency through ribosome display

DOE Office of Scientific and Technical Information (OSTI.GOV)

Mie, Masayasu; Shimizu, Shun; Takahashi, Fumio

2008-08-15

The 5'-untranslated region (5'-UTR) of mRNAs functions as a translation enhancer, promoting translation efficiency. Many in vitro translation systems exhibit a reduced efficiency in protein translation due to decreased translation initiation. The use of a 5'-UTR sequence with high translation efficiency greatly enhances protein production in these systems. In this study, we have developed an in vitro selection system that favors 5'-UTRs with high translation efficiency using a ribosome display technique. A 5'-UTR random library, comprised of 5'-UTRs tagged with a His-tag and Renilla luciferase (R-luc) fusion, were in vitro translated in rabbit reticulocytes. By limiting the translation period, onlymore » mRNAs with high translation efficiency were translated. During translation, mRNA, ribosome and translated R-luc with His-tag formed ternary complexes. They were collected with translated His-tag using Ni-particles. Extracted mRNA from ternary complex was amplified using RT-PCR and sequenced. Finally, 5'-UTR with high translation efficiency was obtained from random 5'-UTR library.« less

Role of Inflammatory Signaling in the Differential Effects of Saturated and Poly-unsaturated Fatty Acids on Peripheral Circadian Clocks.

PubMed

Kim, Sam-Moon; Neuendorff, Nichole; Chapkin, Robert S; Earnest, David J

2016-05-01

Inflammatory signaling may play a role in high-fat diet (HFD)-related circadian clock disturbances that contribute to systemic metabolic dysregulation. Therefore, palmitate, the prevalent proinflammatory saturated fatty acid (SFA) in HFD and the anti-inflammatory, poly-unsaturated fatty acid (PUFA), docosahexaenoic acid (DHA), were analyzed for effects on circadian timekeeping and inflammatory responses in peripheral clocks. Prolonged palmitate, but not DHA, exposure increased the period of fibroblast Bmal1-dLuc rhythms. Acute palmitate treatment produced phase shifts of the Bmal1-dLuc rhythm that were larger in amplitude as compared to DHA. These phase-shifting effects were time-dependent and contemporaneous with rhythmic changes in palmitate-induced inflammatory responses. Fibroblast and differentiated adipocyte clocks exhibited cell-specific differences in the time-dependent nature of palmitate-induced shifts and inflammation. DHA and other inhibitors of inflammatory signaling (AICAR, cardamonin) repressed palmitate-induced proinflammatory responses and phase shifts of the fibroblast clock, suggesting that SFA-mediated inflammatory signaling may feed back to modulate circadian timekeeping in peripheral clocks. Copyright © 2016 The Authors. Published by Elsevier B.V. All rights reserved.

The MBD7 complex promotes expression of methylated transgenes without significantly altering their methylation status

PubMed Central

Li, Dongming; Palanca, Ana Marie S; Won, So Youn; Gao, Lei; Feng, Ying; Vashisht, Ajay A; Liu, Li; Zhao, Yuanyuan; Liu, Xigang; Wu, Xiuyun; Li, Shaofang; Le, Brandon; Kim, Yun Ju; Yang, Guodong; Li, Shengben; Liu, Jinyuan; Wohlschlegel, James A; Guo, Hongwei; Mo, Beixin; Chen, Xuemei; Law, Julie A

2017-01-01

DNA methylation is associated with gene silencing in eukaryotic organisms. Although pathways controlling the establishment, maintenance and removal of DNA methylation are known, relatively little is understood about how DNA methylation influences gene expression. Here we identified a METHYL-CpG-BINDING DOMAIN 7 (MBD7) complex in Arabidopsis thaliana that suppresses the transcriptional silencing of two LUCIFERASE (LUC) reporters via a mechanism that is largely downstream of DNA methylation. Although mutations in components of the MBD7 complex resulted in modest increases in DNA methylation concomitant with decreased LUC expression, we found that these hyper-methylation and gene expression phenotypes can be genetically uncoupled. This finding, along with genome-wide profiling experiments showing minimal changes in DNA methylation upon disruption of the MBD7 complex, places the MBD7 complex amongst a small number of factors acting downstream of DNA methylation. This complex, however, is unique as it functions to suppress, rather than enforce, DNA methylation-mediated gene silencing. DOI: http://dx.doi.org/10.7554/eLife.19893.001 PMID:28452714

LUC-2-1682 Anthony Wayne Bridge : main cable long term health monitoring.

DOT National Transportation Integrated Search

2014-01-01

In preparation for a large rehabilitation project on the Anthony Wayne Bridge (AWB), the Ohio Department of : Transportation has expressed interest in evaluating monitoring and protection strategies which may extend the life : of the AWB. This study ...

77 FR 50510 - Federal Agency Responses to Interagency Coordinating Committee on the Validation of Alternative...

Federal Register 2010, 2011, 2012, 2013, 2014

2012-08-21

... methods for potential use in the EPA EDSP. The evaluation indicated that no in vitro ER- or AR-based test... considered a high priority based on the lack of adequately validated test methods and the regulatory and... Limitations of the LUMI-CELL[supreg] ER (BG1Luc ER TA) Test Method, An In Vitro Assay for Identifying Human...

Effects of land use/cover change and harvests on forest carbon dynamics in northern states of the United States from remote sensing and inventory data: 1992-2001

Treesearch

Daolan Zheng; Linda S. Heath; Mark J. Ducey; James E. Smith

2011-01-01

We examined spatial patterns of changes in forest area and nonsoil carbon (C) dynamics affected by land use/cover change (LUC) and harvests in 24 northern states of the United States using an integrated methodology combining remote sensing and ground inventory data between 1992 and 2001. We used the Retrofit Change Product from the Multi-Resolution Land Characteristics...

Happy 40th Cinema Guild

ERIC Educational Resources Information Center

Kuzyk, Raya

2008-01-01

On May 10, 1968, the Cannes Film Festival shuddered to a stop when filmmakers including Jean-Luc Godard and Francois Truffaut rushed the stage at the Palais des Festivals to protest French New Waver Henri Langlois's removal from his post as president of the National Cinematheque Francaise. Also that month, in that same spirit of film lovers'…

Design of the subject of quality engineering and security of the product of the degree in engineering in industrial design and development of product based in the methodology of the case

NASA Astrophysics Data System (ADS)

González, M. R.; Lambán, M. P.

2012-04-01

This paper presents the result of designing the subject Quality Engineering and Security of the Product, belonging to the Degree of Engineering in Industrial Design and Product Development, on the basis of the case methodology. Practical sessions of this subject are organized using the whole documents of the Quality System Management of the virtual company BeaLuc S.A.

Figures of Disengagement: Charles Taylor, Scientific Parenting, and the Paradox of Late Modernity

ERIC Educational Resources Information Center

Van den Berge, Luc; Ramaekers, Stefan

2014-01-01

In this essay Luc Van den Berge and Stefan Ramaekers take the idea(l) of "scientific parenting" as an example of ambiguities that are typical of our late-modern condition. On the one hand, parenting seems like a natural thing to do, which makes "scientific parenting" sound like an oxymoron; on the other hand, a disengaged…

U.S. Naval Forces, Vietnam Monthly Historical Summary for October 1969

DTIC Science & Technology

1969-12-12

killed in the action, and there were no friendly casualties with the exception of a helo that was hit by « neny fire forcing it to return to Ben Luc. j...information and captured documents were particularly grat- ifying in that they pointed to the extreme difficulty that the « neny «as having in moving food

Reviews and syntheses: Soil N2O and NO emissions from land use and land-use change in the tropics and subtropics: a meta-analysis

NASA Astrophysics Data System (ADS)

van Lent, J.; Hergoualc'h, K.; Verchot, L. V.

2015-12-01

Deforestation and forest degradation in the tropics may substantially alter soil N-oxide emissions. It is particularly relevant to accurately quantify those changes to properly account for them in a REDD+ climate change mitigation scheme that provides financial incentives to reduce the emissions. With this study we provide updated land use (LU)-based emission rates (104 studies, 392 N2O and 111 NO case studies), we determine the trend and magnitude of flux changes with land-use change (LUC) using a meta-analysis approach (44 studies, 135 N2O and 37 NO cases) and evaluate biophysical drivers of N2O and NO emissions and emission changes for the tropics. The average N2O and NO emissions in intact upland tropical forest amounted to 2.0 ± 0.2 (n = 90) and 1.7 ± 0.5 (n = 36) kg N ha-1 yr-1, respectively. In agricultural soils annual N2O emissions were exponentially related to N fertilization rates and average water-filled pore space (WFPS) whereas in non-agricultural sites a Gaussian response to WFPS fit better with the observed NO and N2O emissions. The sum of soil N2O and NO fluxes and the ratio of N2O to NO increased exponentially and significantly with increasing nitrogen availability (expressed as NO3- / [NO3-+NH4+]) and WFPS, respectively; following the conceptual Hole-In-the-Pipe model. Nitrous and nitric oxide fluxes did not increase significantly overall as a result of LUC (Hedges's d of 0.11 ± 0.11 and 0.16 ± 0.19, respectively), however individual LUC trajectories or practices did. Nitrous oxide fluxes increased significantly after intact upland forest conversion to croplands (Hedges's d = 0.78 ± 0.24) and NO increased significantly following the conversion of low forest cover (secondary forest younger than 30 years, woodlands, shrublands) (Hedges's d of 0.44 ± 0.13). Forest conversion to fertilized systems significantly and highly raised both N2O and NO emission rates (Hedges's d of 1.03 ± 0.23 and 0.52 ± 0.09, respectively). Changes in nitrogen

Soil N2O and NO emissions from land use and land-use change in the tropics and subtropics: a meta-analysis

NASA Astrophysics Data System (ADS)

van Lent, J.; Hergoualc'h, K.; Verchot, L. V.

2015-08-01

Deforestation and forest degradation in the tropics may substantially alter soil N-oxide emissions. It is particularly relevant to accurately quantify those changes to properly account for them in a REDD+ climate change mitigation scheme that provides financial incentives to reduce the emissions. With this study we provide updated land use (LU)-based emission rates (103 studies, 387 N2O and 111 NO case studies), determine the trend and magnitude of flux changes with land-use change (LUC) using a meta-analysis approach (43 studies, 132 N2O and 37 NO cases) and evaluate biophysical drivers of N2O and NO emissions and emission changes for the tropics. The average N2O and NO emissions in intact upland tropical forest amounted to 2.0 ± 0.2 (n = 88) and 1.7 ± 0.5 (n = 36) kg N ha-1 yr-1, respectively. In agricultural soils annual N2O emissions were exponentially related to N fertilization rates and average water-filled pore space (WFPS) whereas in non-agricultural sites a Gaussian response to WFPS fit better the observed NO and N2O emissions. The sum of soil N2O and NO fluxes and the ratio of N2O to NO increased exponentially and significantly with increasing nitrogen availability (expressed as NO3-/[NO3-+NH4+]) and WFPS, respectively; following the conceptual Hole-In-the-Pipe model. Nitrous and nitric oxide fluxes did not overall increase significantly as a result of LUC (Hedges's d of 0.11 ± 0.11 and 0.16 ± 0.19, respectively), however individual LUC trajectories or practices did. Nitrous oxide fluxes increased significantly after intact upland forest conversion to croplands (Hedges's d = 0.78 ± 0.24) and NO increased significantly following the conversion of low forest cover (secondary forest younger than 30 years, woodlands, shrublands) (Hedges's d of 0.44 ± 0.13). Forest conversion to fertilized systems significantly and highly raised both N2O and NO emission rates (Hedges's d of 1.03 ± 0.23 and 0.52 ± 0.09, respectively). Changes in nitrogen availability

Cosmic Perspectives

NASA Astrophysics Data System (ADS)

Biswas, S. K.; Mallik, D. C. V.; Vishveshwara, C. V.

2008-07-01

1. Astronomy in ancient and medieval China Joseph Needham; 2. Indian astronomy: an historical perspective B. V. Subbarayappa; 3. Making of astronomy in ancient India Debiprasad Chattopadhyaya; 4. The impact of astronomy on the development of western science Jean-Claude Pecker; 5. Man and the Universe Hubert Reeves; 6. Understanding the Universe - challenges and directions in modern observational astronomy Harlan Smith, Jr: 7. Frontiers in cosmology Fred Hoyle; 8. Did the Universe originate in a big bang? Jayant Narlikar; 9. The dark matter problem Bernard Carr; 10. Geometry and the Universe C. V. Vishveshwara; 11. The origin and evolution of life Cyril Ponnamperuma; 12. The anthropic principle: self selection as an adjunct to natural selection Brandon Carter; 13. Astrology and science: an examination of the evidence Ivan Kelly, Roger Culver and Peter Loptson; 14. Astronomy and science fiction Allen Janis.

Unattainable extended spacetime regions in conformal gravity

NASA Astrophysics Data System (ADS)

Chakrabarty, Hrishikesh; Benavides-Gallego, Carlos A.; Bambi, Cosimo; Modesto, Leonardo

2018-03-01

The Janis-Newman-Winicour metric is a solution of Einstein's gravity minimally coupled to a real massless scalar field. The γ-metric is instead a vacuum solution of Einstein's gravity. Both spacetimes have no horizon and possess a naked singularity at a finite value of the radial coordinate, where curvature invariants diverge and the spacetimes are geodetically incomplete. In this paper, we reconsider these solutions in the framework of conformal gravity and we show that it is possible to solve the spacetime singularities with a suitable choice of the conformal factor. Now curvature invariants remain finite over the whole spacetime. Massive particles never reach the previous singular surface and massless particles can never do it with a finite value of their affine parameter. Our results support the conjecture according to which conformal gravity can fix the singularity problem that plagues Einstein's gravity.

Self-esteem and obsessive compulsive disorder.

PubMed

Husain, Nusrat; Chaudhry, Imran; Raza-ur-Rehman; Ahmed, Ghazal Riaz

2014-01-01

To explore the association between self-esteem and obsessive compulsive disorder in a low-income country, and to conduct an in-depth analysis into the said relationship by identifying any confounding variables that might exist. The cross-sectional study was conducted at the psychiatry out-patient clinic of Civil Hospital, Karachi, from January to March 2008, and comprised 65 patients diagnosed with obsessive compulsive disorder and 30 healthy controls. The participatnts completed the Janis and Field Social Adequacy scale and the Rosenberg Self-esteem scale. SPSS 15 was used for statistical analysis. Significantly different scores were reported on both measures of self-esteem between the patients and the controls (p<0.001 each), indicating reduced levels of self-esteem in the patients compared to the controls. Data replicated earlier findings from populations in high-income countries.

Carbon Calculator for Land Use Change from Biofuels Production (CCLUB). Users' Manual and Technical Documentation

DOE Office of Scientific and Technical Information (OSTI.GOV)

Dunn, Jennifer B.; Qin, Zhangcai; Mueller, Steffen

The Carbon Calculator for Land Use Change from Biofuels Production (CCLUB) calculates carbon emissions from land use change (LUC) for four different ethanol production pathways including corn grain ethanol and cellulosic ethanol from corn stover, Miscanthus, and switchgrass. This document discusses the version of CCLUB released September 30, 2014 which includes corn and three cellulosic feedstocks: corn stover, Miscanthus, and switchgrass.

Pooling procurement in the Belgian hospital sector.

PubMed

Hebert, Guy

2011-01-01

The Belgian hospital sector is following the example of a number of other European countries and for more than ten years now, has been striving to pool its medical supplies and equipment purchases in a bid to reduce costs. The various experiments of which we are aware come under both opportunist purchases and initiatives which are designed to encourage local-regional contracts. These attempts have now all come to nothing or are struggling in the absence of a structured and professional approach. In 2005, the Saint Luc University Clinic in Brussels decided to set up a high-performance purchasing department, the aim being to centre its initiatives around TCO or Total Cost of Ownership. Following an analysis of the various experiments into pooling procurement in hospitals in Europe, the Saint Luc University Clinic decided on a central procurement agency model, in accordance with new legislation on public procurement. This article seeks to highlight the prerequisites which are vital for a procurement pooling initiative, without underestimating the risks and limitations of implementing such a change in procurement practices. The Mercure central procurement agency is now the largest interhospital purchasing structure in Belgium.

Butyric acid stimulates bone sialoprotein gene transcription.

PubMed

Yang, Li; Li, Zhengyang; Li, Xinyue; Wang, Zhitao; Wang, Shuang; Sasaki, Yoko; Takai, Hideki; Ogata, Yorimasa

2010-06-01

Butyric acid (sodium butyrate; BA) is an extracellular metabolite secreted from periodontopathic bacteria present in subgingival plaque. BA induces apoptosis of T and B cells, and acts as a potent inhibitor of histone deacetylases. Bone sialoprotein (BSP) is thought to function in the initial mineralization of bone, and may be crucial for osteoblast differentiation, bone matrix mineralization and tumor metastasis. In the present study we investigated the regulation of BSP transcription by BA in rat osteoblast-like ROS17/2.8 cells. At 12 h, BA (10(-4) M) increased the level of BSP mRNA, and enhanced the luciferase activity of the construct pLUC3, which includes the promoter sequence between nucleotides -116 and +60. Transcriptional stimulation by BA was abrogated in the pLUC3 construct which containing a 2-bp mutation in the fibroblast growth factor 2 response element (FRE). Gel shift analyses showed that BA increased the binding of nuclear protein to FRE. These data suggest that BA increases the transcription of the BSP gene mediated through FRE in the rat BSP gene promoter, and induces osteoblast activity in the early stage of bone formation.

Transgenic mouse model harboring the transcriptional fusion ccl20-luciferase as a novel reporter of pro-inflammatory response.

PubMed

Crispo, Martina; Van Maele, Laurye; Tabareau, Julien; Cayet, Delphine; Errea, Agustina; Ferreira, Ana María; Rumbo, Martin; Sirard, Jean Claude

2013-01-01

The chemokine CCL20, the unique ligand of CCR6 functions as an attractant of immune cells. Expression of CCL20 is induced by Toll-like Receptor (TLR) signaling or proinflammatory cytokine stimulation. However CCL20 is also constitutively produced at specific epithelial sites of mucosa. This expression profile is achieved by transcriptional regulation. In the present work we characterized regulatory features of mouse Ccl20 gene. Transcriptional fusions between the mouse Ccl20 promoter and the firefly luciferase (luc) encoding gene were constructed and assessed in in vitro and in vivo assays. We found that liver CCL20 expression and luciferase activity were upregulated by systemic administration of the TLR5 agonist flagellin. Using shRNA and dominant negative form specific for mouse TLR5, we showed that this expression was controlled by TLR5. To address in situ the regulation of gene activity, a transgenic mouse line harboring a functional Ccl20-luc fusion was generated. The luciferase expression was highly concordant with Ccl20 expression in different tissues. Our data indicate that the transgenic mouse model can be used to monitor activation of innate response in vivo.

In vitro OECD test methods applied to screen the estrogenic effect of chemicals, used in Korea.

PubMed

Lee, Hee-Seok; Park, Eun-Jung; Han, Songyi; Oh, Gyeong-Yong; Kim, Min-Hee; Kang, Hui-Seung; Suh, Jin-Hyang; Oh, Jae-Ho; Lee, Kwang-Soo; Hwang, Myung-Sil; Moon, Guiim; Hong, Jin-Hwan; Hwang, In-Gyun

2016-09-01

In this study, 27 chemicals found in household products, which became an issue in Korea were screened for the agonistoc and antagonistic effects against human estrogen receptor using official Organization for Economic Cooperation and Development (OECD) in vitro assays, STTA assay using ERα-HeLa-9903 cell line and BG1Luc ER TA assay. In the case of human ER agonist screening by two assays, all tested chemicals did not show agonist effect against ER. In ER antagonist test by BG1Luc ER TA assay, five surfactants α-dodecyl-ω-hydroxypoly(oxyethylene), alcohols C16-18 ethoxylated, nonylphenol, ethoxylated, 3,6,9,12,15,18,21-heptaoxatritriacontan-1-ol, and α-dodecyl-ω-hydroxypoly(oxy-1,2-ethanediyl)) were found to exhibit weak antagonistic activities. The agonist/antagonist effects against human estrogen receptor of various chemicals, used in Korea by OECD test guideline are reported in this study. These results indicated that two OECD in vitro assays will can be applied in Korea by screening of agonistic/antagonistic effects against human ER of various chemicals. Copyright © 2016 Elsevier Ltd. All rights reserved.

In vivo gene delivery and expression by bacteriophage lambda vectors.

PubMed

Lankes, H A; Zanghi, C N; Santos, K; Capella, C; Duke, C M P; Dewhurst, S

2007-05-01

Bacteriophage vectors have potential as gene transfer and vaccine delivery vectors because of their low cost, safety and physical stability. However, little is known concerning phage-mediated gene transfer in mammalian hosts. We therefore performed experiments to examine phage-mediated gene transfer in vivo. Mice were inoculated with recombinant lambda phage containing a mammalian expression cassette encoding firefly luciferase (luc). Efficient, dose-dependent in vivo luc expression was detected, which peaked within 24 h of delivery and declined to undetectable levels within a week. Display of an integrin-binding peptide increased cellular internalization of phage in vitro and enhanced phage-mediated gene transfer in vivo. Finally, in vivo depletion of phagocytic cells using clodronate liposomes had only a minor effect on the efficiency of phage-mediated gene transfer. Unmodified lambda phage particles are capable of transducing mammalian cells in vivo, and may be taken up -- at least in part -- by nonphagocytic mechanisms. Surface modifications that enhance phage uptake result in more efficient in vivo gene transfer. These experiments shed light on the mechanisms involved in phage-mediated gene transfer in vivo, and suggest new approaches that may enhance the efficiency of this process.

Composite three-layer closure of oral antral communication with 10 months follow-up-a case study.

PubMed

Weinstock, Robert J; Nikoyan, Levon; Dym, Harry

2014-02-01

We propose a 3-layer composite closure technique for an oral antral communication (OAC) while avoiding secondary donor site morbidity. A patient had developed a 1-cm OAC after extraction of right maxillary first molar. The patient subsequently developed acute maxillary sinusitis. The patient was taken to the operating room, and a Caldwell-Luc procedure was performed. The bony window from the Caldwell-Luc was "press fit" over the bony OAC defect. Soft tissue closure was then achieved with a buccal fat pad flap and a buccal mucosal advancement flap. The patient was examined on postoperative day 5 and 1, 2, 3, 6, and 10 months postoperatively. The acute sinusitis had resolved. The soft tissue closure was successful. The bone graft remained intact, prevented sinus pneumatization, and restored continuity to the floor of the maxillary sinus. The presented technique for 3-layer closure of OACs allows for the stability of a double-layer closure of OAC with the added benefit of bone grafting from single operative site, achieving stable oral antral closure, bone grafting, and the avoidance of secondary donor site morbidity. Copyright © 2014 American Association of Oral and Maxillofacial Surgeons. Published by Elsevier Inc. All rights reserved.

Assessing the potential impact and uncertainty of climate, land use change and demographic trends on malaria transmission in Africa by 2050.

NASA Astrophysics Data System (ADS)

Tompkins, Adrian; Caporaso, Luca; Colon-Gonzalez, Felipe

2014-05-01

Previous analyses of data has shown that in addition to variability and longer term trends in climate variables, both land use change (LUC) and population mobility and urbanisation trends can impact malaria transmission intensities and socio-economic burden. With the new regional VECTRI dynamical malaria model it is now possible to examine these in an integrated modelling framework. Using 5 global climate models which were bias corrected using the WATCH data for the recent ISIMIP project, the four Representative Concentration Pathways (RCP), population projections disaggregated from the Shared Socioeconomic Pathways (SSP) and Land use change from the HYDE model output used in the CMIP5 process, we construct a multi-member ensemble of malaria transmission intensity projections for 2050. The ensemble integrations indicate that climate has the leading impact on malaria changes, but that population growth and urbanisation can offset the effect of climate locally. LUC impacts can also be significant on the local scale but their assessment is highly uncertain and only indicative in this study. It is argued that the study should be repeated with a range of malaria models or VECTRI configurations in order to assess the additional uncertainty due to the malaria model assumptions.

Carbachol Induces Phase-dependent Phase Shifts of Per1 Transcription Rhythms in Cultured Suprachiasmatic Nucleus Slices.

PubMed

Dojo, Kumiko; Yamaguchi, Yoshiaki; Fustin, Jean-Michel; Doi, Masao; Kobayashi, Masaki; Okamura, Hitoshi

2017-04-01

Among nonphotic stimulants, a classic cholinergic agonist, carbachol, is known to have a strong and unique phase-resetting effect on the circadian clock: Intracerebroventricular carbachol treatment causes phase delays during the subjective early night and phase advances in the subjective late night, but the effects of this drug on the suprachiasmatic nucleus (SCN) in vivo and in vitro are still controversial. In the present study, we succeeded in reproducing the biphasic phase-shifting effect of carbachol on clock gene expression in organotypic SCN slices prepared from mice carrying a Per1-promoter fused luciferase gene ( Per1-luc). Since this biphasic effect of carbachol in Per1-luc SCN was prevented by atropine but not by mecamylamine, we concluded that these phase shifts were muscarinic receptor-dependent. Next, we analyzed the expression of muscarinic receptors in the SCN by in situ hybridization and found that M3 and M4 subtypes were expressed in SCN cells. These signals appeared neonatally and reached adult levels at postnatal day 10. Together, these findings suggest that carbachol has a phase-dependent phase-shifting effect on the SCN clock through muscarinic receptor subtypes expressed in the SCN.

Transgenic Mouse Model Harboring the Transcriptional Fusion Ccl20-Luciferase as a Novel Reporter of Pro-Inflammatory Response

PubMed Central

Crispo, Martina; Van Maele, Laurye; Tabareau, Julien; Cayet, Delphine; Errea, Agustina; Ferreira, Ana María; Rumbo, Martin; Sirard, Jean Claude

2013-01-01

The chemokine CCL20, the unique ligand of CCR6 functions as an attractant of immune cells. Expression of CCL20 is induced by Toll-like Receptor (TLR) signaling or proinflammatory cytokine stimulation. However CCL20 is also constitutively produced at specific epithelial sites of mucosa. This expression profile is achieved by transcriptional regulation. In the present work we characterized regulatory features of mouse Ccl20 gene. Transcriptional fusions between the mouse Ccl20 promoter and the firefly luciferase (luc) encoding gene were constructed and assessed in in vitro and in vivo assays. We found that liver CCL20 expression and luciferase activity were upregulated by systemic administration of the TLR5 agonist flagellin. Using shRNA and dominant negative form specific for mouse TLR5, we showed that this expression was controlled by TLR5. To address in situ the regulation of gene activity, a transgenic mouse line harboring a functional Ccl20-luc fusion was generated. The luciferase expression was highly concordant with Ccl20 expression in different tissues. Our data indicate that the transgenic mouse model can be used to monitor activation of innate response in vivo. PMID:24265691

Luciferase-tagged wild-type and tropism-deficient mouse cytomegaloviruses reveal early dynamics of host colonization following peripheral challenge.

PubMed

Farrell, Helen; Oliveira, Martha; Macdonald, Kate; Yunis, Joseph; Mach, Michael; Bruce, Kimberley; Stevenson, Philip; Cardin, Rhonda; Davis-Poynter, Nicholas

2016-12-01

Cytomegaloviruses (CMVs) establish persistent, systemic infections and cause disease by maternal-foetal transfer, suggesting that their dissemination is a key target for antiviral intervention. Late clinical presentation has meant that human CMV (HCMV) dissemination is not well understood. Murine CMV (MCMV) provides a tractable model. Whole mouse imaging of virus-expressed luciferase has proved a useful way to track systemic infections. MCMV, in which the abundant lytic gene M78 was luciferase-tagged via a self-cleaving peptide (M78-LUC), allowed serial, unbiased imaging of systemic and peripheral infection without significant virus attenuation. Ex vivo luciferase imaging showed greater sensitivity than plaque assay, and revealed both well-known infection sites (the lungs, lymph nodes, salivary glands, liver, spleen and pancreas) and less explored sites (the bone marrow and upper respiratory tract). We applied luciferase imaging to tracking MCMV lacking M33, a chemokine receptor conserved in HCMV and a proposed anti-viral drug target. M33-deficient M78-LUC colonized normally in peripheral sites and local draining lymph nodes but spread poorly to the salivary gland, suggesting a defect in vascular transport consistent with properties of a chemokine receptor.

REPRESSOR OF SILENCING5 Encodes a Member of the Small Heat Shock Protein Family and Is Required for DNA Demethylation in Arabidopsis[C][W

PubMed Central

Zhao, Yusheng; Xie, Shaojun; Li, Xiaojie; Wang, Chunlei; Chen, Zhongzhou; Lai, Jinsheng; Gong, Zhizhong

2014-01-01

In Arabidopsis thaliana, active DNA demethylation is initiated by the DNA glycosylase REPRESSOR OF SILENCING1 (ROS1) and its paralogs DEMETER, DEMETER-LIKE2 (DML2), and DML3. How these demethylation enzymes are regulated, however, is poorly understood. Here, using a transgenic Arabidopsis line harboring the stress-inducible RESPONSIVE TO DEHYDRATION29A (RD29A) promoter–LUCIFERASE (LUC) reporter gene and the cauliflower mosaic virus 35S promoter (35S)–NEOMYCIN PHOSPHOTRANSFERASE II (NPTII) antibiotic resistance marker gene, we characterize a ROS locus, ROS5, that encodes a protein in the small heat shock protein family. ROS5 mutations lead to the silencing of the 35S-NPTII transgene due to DNA hypermethylation but do not affect the expression of the RD29A-LUC transgene. ROS5 physically interacts with the histone acetyltransferase ROS4/INCREASED DNA METHYLATION1 (IDM1) and is required to prevent the DNA hypermethylation of some genes that are also regulated by ROS1 and IDM1. We propose that ROS5 regulates DNA demethylation by interacting with IDM1, thereby creating a chromatin environment that facilitates the binding of ROS1 to erase DNA methylation. PMID:24920332

Epinephrine-Induced and Antiapoptotic Signaling in Prostate Cancer

DTIC Science & Technology

2009-05-01

Phosphorylation of ectopically expressed HA-BAD mirrors endogenous BAD phosphorylation, thus analysis of HA-BAD allows adequately interpret modifications...subjected to emotional stress (Fig.2 A). However, more extensive analysis of CREB phosphorylation in C42Luc xenografts showed that in some cases...data indicate that assignment of tumor samples for analysis of signaling pathways should be done based on blood epinephrine measurements. Given

Test Area C-64 Range Environmental Assessment, Revision 1

DTIC Science & Technology

2010-10-01

DOI U.S. Department of the Interior DNL Day–Night Average Sound Level DU Depleted Uranium EBD Environmental Baseline Document EIAP Environmental...vulnerability, burning sensitivity, drop tests, bullet impact tests, sympathetic detonation tests, advanced warhead design tests, and depleted uranium (DU...land back to range use. Source: U.S. Air Force, 2009 DU = depleted uranium ; ERP = Environmental Restoration Program; LUC = land use control; RW

Endoscopic graduated multiangle, multicorridor resection of juvenile nasopharyngeal angiofibroma: an individualized, tailored, multicorridor skull base approach.

PubMed

Liu, James K; Husain, Qasim; Kanumuri, Vivek; Khan, Mohemmed N; Mendelson, Zachary S; Eloy, Jean Anderson

2016-05-01

OBJECT Juvenile nasopharyngeal angiofibromas (JNAs) are formidable tumors because of their hypervascularity and difficult location in the skull base. Traditional transfacial procedures do not always afford optimal visualization and illumination, resulting in significant morbidity and poor cosmesis. The advent of endoscopic procedures has allowed for resection of JNAs with greater surgical freedom and decreased incidence of facial deformity and scarring. METHODS This report describes a graduated multiangle, multicorridor, endoscopic approach to JNAs that is illustrated in 4 patients, each with a different tumor location and extent. Four different surgical corridors in varying combinations were used to resect JNAs, based on tumor size and location, including an ipsilateral endonasal approach (uninostril); a contralateral, transseptal approach (binostril); a sublabial, transmaxillary Caldwell-Luc approach; and an orbitozygomatic, extradural, transcavernous, infratemporal fossa approach (transcranial). One patient underwent resection via an ipsilateral endonasal uninostril approach (Corridor 1) only. One patient underwent a binostril approach that included an additional contralateral transseptal approach (Corridors 1 and 2). One patient underwent a binostril approach with an additional sublabial Caldwell-Luc approach for lateral extension in the infratemporal fossa (Corridors 1-3). One patient underwent a combined transcranial and endoscopic endonasal/sublabial Caldwell-Luc approach (Corridors 1-4) for an extensive JNA involving both the lateral infratemporal fossa and cavernous sinus. RESULTS A graduated multiangle, multicorridor approach was used in a stepwise fashion to allow for maximal surgical exposure and maneuverability for resection of JNAs. Gross-total resection was achieved in all 4 patients. One patient had a postoperative CSF leak that was successfully repaired endoscopically. One patient had a delayed local recurrence that was successfully resected

In vivo characterisation of the inflammatory reaction following mesh implantation in transgenic mice models.

PubMed

Fet, N; Alizai, P H; Fragoulis, A; Wruck, C; Pufe, T; Tolba, R H; Neumann, U P; Klinge, U

2014-06-01

Hernia repair with prosthetic meshes represents one of the most common surgical procedures in the field of surgery. This intervention is always associated with an ensuing inflammatory response, angiogenesis and fibrotic encapsulation forming a foreign body granuloma (FBG) around the mesh fibres. Several studies have described this inflammatory reaction by characterising inflammatory cell infiltrate around the FBG after mesh explantation. However, very little is known about the real-time progression of such an inflammatory response. The aim of this study was to investigate the feasibility of monitoring the ongoing inflammatory response to mesh implantation using bioluminescence in vivo. Three luciferase transgenic mice strains (FVB/N-Tg(Vegfr2-luc)-Xen, BALB/C-Tg(NFκB-RE-luc)-Xen and Tg(INS/EpRE-Luc)T20Rbl) were used. Mice were anaesthetized with 2 % isoflurane, and two incisions were made on the left and right sides of the abdomen of the mice. A 1-cm(2) propylene mesh was implanted subcutaneously in the right incision wound of each mouse, and the left wound served as control. Two hundred microliters of D-luciferin was injected into the mice, and bioluminescence measurements were done prior to the surgical intervention and subsequently every 3 days. After mesh explantation, histological analysis was done. Statistical analysis was done using prism GraphPad software. Bioluminescence results revealed different time points of maximum signal for the different mice strains. VEGFR2 gene expression peaked on day 6, NFkB on day 12 and ARE on day 3 post mesh implantation. We also observed much higher bioluminescent signal around the FBG surrounding the mesh as compared to the control wound, with p < 0.05 for all the different mice strains. Our results prove the possibility of monitoring the inflammatory reaction after mesh implantation in vivo using bioluminescence signal release. This provides a novel method of accessing and accurately describing the ongoing

Molecular Imaging of Smoke-Induced Changes in Nuclear Factor-Kappa B Expression in Murine Tissues Including the Lung.

PubMed

Syrkina, Olga; Hales, Charles H; Bonab, Ali A; Hamrahi, Victoria; Paul, Kasie; Jung, Walter J; Tompkins, Ronald G; Fischman, Alan J; Carter, Edward A

Many inflammatory responses are mediated by activation of the transcription factor, nuclear factor-kappa B (NF-κB), and a wide variety of human diseases involve abnormal regulation of its expression. In this investigation, we evaluated the effect of smoke inhalation injury on NF-κB expression in lung using two strains of NF-κB reporter mice. Groups of reporter mice with viral thymidine kinase (TK) or "fire fly" luciferase (Luc) genes under control by the NF-κB promoter (TK/NF-κB mice and Luc/NF-κB mice) were subjected to nonlethal smoke inhalation injury. Sham-treated animals served as controls. Twenty-four hours (each animal was injected intravenously with either 9-(4-18F-fluoro-3-[hydroxymethyl]butyl)guanine (FHBG) (~ 1.0 mCi) or luciferin (1.0 mg). One hour later, the TK/NF-κB mice were studied by micro-positron emission tomography (µ-PET) imaging using a Concord P4 µ-PET camera, and the Luc/NF-κB mice were studied by bioluminescence imaging with a charge-coupled device camera. The µ-PET data demonstrated that smoke injury produced massive increases in NF-κB expression (FHBG-standardized uptake value: 3.1 vs 0.0) 24 hours after smoke inhalation, which was reduced 48 hours after smoke inhalation, but still significantly different than the control. Qualitative analysis of the bioluminescence data revealed a remarkably similar effect of burn NF-κB luciferase expression in vivo. Biodistribution studies of FHBG uptake and luciferase activity in lung tissue demonstrated a similar increase 24 hours after injury, which was reduced 48 hours later, but still significantly higher than the sham. The present data with these models providing longitudinal imaging data on the same mouse may prove useful in the examination of the factors producing lung injury by smoke inhalation, as well as the treatment(s) for the damage produced with and without burn injury.

Generation of SNCA Cell Models Using Zinc Finger Nuclease (ZFN) Technology for Efficient High-Throughput Drug Screening.

PubMed

Dansithong, Warunee; Paul, Sharan; Scoles, Daniel R; Pulst, Stefan M; Huynh, Duong P

2015-01-01

Parkinson's disease (PD) is a progressive neurodegenerative disorder caused by loss of dopaminergic neurons of the substantia nigra. The hallmark of PD is the appearance of neuronal protein aggregations known as Lewy bodies and Lewy neurites, of which α-synuclein forms a major component. Familial PD is rare and is associated with missense mutations of the SNCA gene or increases in gene copy number resulting in SNCA overexpression. This suggests that lowering SNCA expression could be therapeutic for PD. Supporting this hypothesis, SNCA reduction was neuroprotective in cell line and rodent PD models. We developed novel cell lines expressing SNCA fused to the reporter genes luciferase (luc) or GFP with the objective to enable high-throughput compound screening (HTS) for small molecules that can lower SNCA expression. Because SNCA expression is likely regulated by far-upstream elements (including the NACP-REP1 located at 8852 bp upstream of the transcription site), we employed zinc finger nuclease (ZFN) genome editing to insert reporter genes in-frame downstream of the SNCA gene in order to retain native SNCA expression control. This ensured full retention of known and unknown up- and downstream genetic elements controlling SNCA expression. Treatment of cells with the histone deacetylase inhibitor valproic acid (VPA) resulted in significantly increased SNCA-luc and SNCA-GFP expression supporting the use of our cell lines for identifying small molecules altering complex modes of expression control. Cells expressing SNCA-luc treated with a luciferase inhibitor or SNCA siRNA resulted in Z'-scores ≥ 0.75, suggesting the suitability of these cell lines for use in HTS. This study presents a novel use of genome editing for the creation of cell lines expressing α-synuclein fusion constructs entirely under native expression control. These cell lines are well suited for HTS for compounds that lower SNCA expression directly or by acting at long-range sites to the SNCA

Modeling urbanized watershed flood response changes with distributed hydrological model: key hydrological processes, parameterization and case studies

NASA Astrophysics Data System (ADS)

Chen, Y.

2017-12-01

Urbanization is the world development trend for the past century, and the developing countries have been experiencing much rapider urbanization in the past decades. Urbanization brings many benefits to human beings, but also causes negative impacts, such as increasing flood risk. Impact of urbanization on flood response has long been observed, but quantitatively studying this effect still faces great challenges. For example, setting up an appropriate hydrological model representing the changed flood responses and determining accurate model parameters are very difficult in the urbanized or urbanizing watershed. In the Pearl River Delta area, rapidest urbanization has been observed in China for the past decades, and dozens of highly urbanized watersheds have been appeared. In this study, a physically based distributed watershed hydrological model, the Liuxihe model is employed and revised to simulate the hydrological processes of the highly urbanized watershed flood in the Pearl River Delta area. A virtual soil type is then defined in the terrain properties dataset, and its runoff production and routing algorithms are added to the Liuxihe model. Based on a parameter sensitive analysis, the key hydrological processes of a highly urbanized watershed is proposed, that provides insight into the hydrological processes and for parameter optimization. Based on the above analysis, the model is set up in the Songmushan watershed where there is hydrological data observation. A model parameter optimization and updating strategy is proposed based on the remotely sensed LUC types, which optimizes model parameters with PSO algorithm and updates them based on the changed LUC types. The model parameters in Songmushan watershed are regionalized at the Pearl River Delta area watersheds based on the LUC types of the other watersheds. A dozen watersheds in the highly urbanized area of Dongguan City in the Pearl River Delta area were studied for the flood response changes due to

Elucidating the mechanism of action of tributyltin (TBT) in zebrafish.

PubMed

McGinnis, Courtney L; Crivello, Joseph F

2011-05-01

Tributyltin (TBT), an antifouling agent, has been implicated in the masculinization of fish species worldwide, but the masculinizing mechanism is not fully understood. We have examined the actions of TBT as an endocrine disruptor in zebrafish (Danio rerio). In HeLa cells transiently co-transfected with plasmid constructs containing the zebrafish estrogen receptors (zfERα, zfERβ(1) and zfERβ(2)) and the zebrafish estrogen response element (zfERE-tk-luc), ethinyl estradiol (EE2) induced luciferase activity 4 to 6-fold and was inhibited by TBT. In HeLa cells transiently co-transfected with the zebrafish androgen receptor (zfAR) and the murine androgen receptor response element (ARE-slp-luc), testosterone induced luciferase activity was not inhibited by TBT. In HeLa cells co-transfected with zfERα, zfERβ(1) and zfERβ(2) and a plasmid containing zebrafish aromatase (zfCyp19b-luc), TBT inhibited luciferase activity. In zebrafish exposed to 1mg/kg and 5mg/kg TBT in vivo, there was a increase in liver sulfotransferase and a decrease acyl-CoA testosterone acyltransferase activity. Real-time PCR analysis of sexual differentiation markers in fish exposed to TBT in vivo revealed a tissue-specific response. In brain there was increased production of Sox9, Dax1, and SF1 mRNA, an androgenizing effect, while in the liver there was increased production of Dax1, Cyp19a and zfERβ(1) mRNA but decreased production of Sox9 mRNA, a feminizing effect. In the gonads there was increased production of zfERα and zfCyp19a mRNA, again a feminizing effect. TBT has an overall masculinizing effect but the masculinizing effect is tempered by a feminizing effect on gene transcription in certain tissues. These results are discussed in the context of TBT as an endocrine disruptor in zebrafish. Copyright © 2011 Elsevier B.V. All rights reserved.

Magnetic single-walled carbon nanotubes as efficient drug delivery nanocarriers in breast cancer murine model: noninvasive monitoring using diffusion-weighted magnetic resonance imaging as sensitive imaging biomarker.

PubMed

Al Faraj, Achraf; Shaik, Abjal Pasha; Shaik, Asma Sultana

2015-01-01

Targeting doxorubicin (DOX) by means of single-walled carbon nanotube (SWCNT) nanocarriers may help improve the clinical utility of this highly active therapeutic agent. Active targeting of SWCNTs using tumor-specific antibody and magnetic attraction by tagging the nanotubes with iron oxide nanoparticles can potentially reduce the unnecessary side effects and provide enhanced theranostics. In the current study, the in vitro and in vivo efficacy of DOX-loaded SWCNTs as theranostic nanoprobes was evaluated in a murine breast cancer model. Iron-tagged SWCNTs conjugated with Endoglin/CD105 antibody with or without DOX were synthetized and extensively characterized. Their biocompatibility was assessed in vitro in luciferase (Luc2)-expressing 4T1 (4T1-Luc2) murine breast cancer cells using TiterTACS™ Colorimetric Apoptosis Detection Kit (apoptosis induction), poly (ADP-ribose) polymerase (marker for DNA damage), and thiobarbituric acid-reactive substances (oxidative stress generation) assays, and the efficacy of DOX-loaded SWCNTs was evaluated by measuring the radiance efficiency using bioluminescence imaging (BLI). Tumor progression and growth were monitored after 4T1-Luc2 cells inoculation using noninvasive BLI and magnetic resonance imaging (MRI) before and after subsequent injection of SWCNT complexes actively and magnetically targeted to tumor sites. Significant increases in apoptosis, DNA damage, and oxidative stress were induced by DOX-loaded SWCNTs. In addition, a tremendous decrease in bioluminescence was observed in a dose- and time-dependent manner. Noninvasive BLI and MRI revealed successful tumor growth and subsequent attenuation along with metastasis inhibition following DOX-loaded SWCNTs injection. Magnetic tagging of SWCNTs was found to produce significant discrepancies in apparent diffusion coefficient values providing a higher contrast to detect treatment-induced variations as noninvasive imaging biomarker. In addition, it allowed their sensitive

Evaluation of a bioluminescent mouse model expressing aromatase PII-promoter-controlled luciferase as a tool for the study of endocrine disrupting chemicals

DOE Office of Scientific and Technical Information (OSTI.GOV)

Rivest, Patricia, E-mail: patricia.rivest@iaf.inrs.ca; Devine, Patrick J., E-mail: patrick.devine@iaf.inrs.ca; Sanderson, J. Thomas, E-mail: thomas.sanderson@iaf.inrs.c

Dysfunction of the enzyme aromatase (CYP19) is associated with endocrine pathologies such as osteoporosis, impaired fertility and development of hormone-dependent cancers. Certain endocrine disrupting chemicals affect aromatase expression and activity in vitro, but little is known about their ability to do so in vivo. We evaluated a bioluminescent mouse model (LPTA (registered)) CD-1-Tg(Cyp19-luc)-Xen) expressing luciferase under control of the gonadal aromatase pII promoter as an in vivo screening tool for chemicals that may affect aromatase expression. We studied the effects of forskolin, pregnant mare serum gonadotropin and atrazine in this model (atrazine was previously shown to induced pII-promoter-driven aromatase expressionmore » in H295R human adrenocortical carcinoma cells). About 2-4 out of every group of 10 male or female Cyp19-luc mice injected i.p. with 10 mg/kg forskolin had increased gonadal bioluminescence after 3-5 days compared to controls; the others appeared non-responsive. Similarly, about 4 per group of 9 individual females injected with pregnant mare serum gonadotropin had increased ovarian bioluminescence after 24 h. There was a statistically significant correlation between ovarian bioluminescence and plasma estradiol concentrations (n = 14; p = 0.022). Males exposed to a single dose of 100 mg/kg or males and females exposed to 5 daily injections of 30 mg/kg atrazine showed no change in gonadal bioluminescence over a 7 day period, but a significant interaction was found between atrazine (100 mg/kg) and time in female mice (p < 0.05; two-way ANOVA). Ex vivo luciferase activity in dissected organs was increased by forskolin in testis, epididymis and ovaries. Atrazine (30 mg/kg/day) increased (30%) luciferase activity significantly in epididymis only. In conclusion, certain individual Cyp19-luc mice are highly responsive to aromatase inducers, suggesting this model, with further optimization, may have potential as an in vivo screening tool

Curcumin induces osteoblast differentiation through mild-endoplasmic reticulum stress-mediated such as BMP2 on osteoblast cells.

PubMed

Son, Hyo-Eun; Kim, Eun-Jung; Jang, Won-Gu

2018-01-15

Curcumin (diferuloylmethane or [1E,6E]-1,7-bis[4-hydroxy-3-methoxyphenyl]-1,6heptadiene-3,5-dione) is a phenolic natural product derived from the rhizomes of the turmeric plant, Curcuma longa. It is reported to have various biological actions such as anti-oxidative, anti-inflammatory, and anti-cancer effects. However, the molecular mechanism of osteoblast differentiation by curcumin has not yet been reported. The cytotoxicity of curcumin was identified using the 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Expression of osteogenic markers and endoplasmic reticulum (ER) stress markers in C3H1-T1/2 cells were measured using reverse-transcriptase polymerase chain reaction (RT-PCR) and Western blotting. Alkaline phosphatase (ALP) staining was performed to assess ALP activity in C3H10T1/2 cells. Transcriptional activity was detected using a luciferase reporter assay. Curcumin increased the expression of genes such as distal-less homeobox 5 (Dlx5), runt-related transcription factor 2 (Runx2), ALP, and osteocalcin (OC), which subsequently induced osteoblast differentiation in C3H10T1/2 cells. In addition, ALP activity and mineralization was found to be increased by curcumin treatment. Curcumin also induced mild ER stress similar to bone morphogenetic protein 2 (BMP2) function in osteoblast cells. Next, we confirmed that curcumin increased mild ER stress and osteoblast differentiation similar to BMP2 in C3H10T1/2 mesenchymal stem cells. Transient transfection studies also showed that curcumin increased ATF6-Luc activity, while decreasing the activities of CREBH-Luc and SMILE-Luc. In addition, similar to BMP2, curcumin induced the phosphorylation of Smad 1/5/9. Overall, these results demonstrate that curcumin-induced mild ER stress increases osteoblast differentiation via ATF6 expression in C3H10T1/2 cells. Copyright © 2017. Published by Elsevier Inc.

Elucidating Mechanisms of Farnesyltransferase Inhibitor Action and Resistance in Breast Cancer by Bioluminescence Imaging

DTIC Science & Technology

2009-06-01

Ras or Cdc42, and a downstream IRES (internal ribosome entry site) to express Renilla luciferase for normalization of infection efficiency. As...internal ribosome entry site (IRES) downstream of Gal4-GFP-VP16-H-Ras/Cdc42 to drive constitutive expression of Renilla luciferase as a means of...for infection efficiency ( renilla luc). 8 mechanisms determining FTI or GGTI sensitivity and resistance in tumors. The system now will be

Testing New Drugs for Treatment of Melanoma Patients Applying Connectivity Map Database Analysis with Melanoma Gene Signatures

DTIC Science & Technology

2012-10-01

use of R packages implemented in Bioconductor. Each dataset was normalized from raw data using the Frozen RMA (fRMA) algorithm . We applied the same...because development of the specific algorithms and fine tuning of the analytic strategy to accomplish this task was not immediately straightforward. We...express firefly luciferase using a retrovirus that encodes a fusion of luciferase and neomycin phosphotransferase (LucNeo), will be implanted and followed

Land use change emissions from oil palm expansion in Pará, Brazil depend on proper policy enforcement on deforested lands

NASA Astrophysics Data System (ADS)

Yui, Sahoko; Yeh, Sonia

2013-12-01

Brazil aims to increase palm oil production to meet the growing national and global demand for edible oil and biodiesel while preserving environmentally and culturally significant areas. As land use change (LUC) is the result of complex interactions between socio-economic and biophysical drivers operating at multiple temporal and spatial scales, the type and location of LUC depend on drivers such as neighboring land use, conversion elasticity, access to infrastructure, distance to markets, and land suitability. The purpose of this study is to develop scenarios to measure the impact of land conversion under three different enforcement scenarios (none, some, and strict enforcement). We found that converting 22.5 million hectares of land can produce approximately 29 billion gallons (110 billion liters) of biodiesel a year. Of that, 22-71% of the area can come from forest land, conservation units, wetland and indigenous areas, emitting 14-84 gCO2e MJ-1. This direct land use emission alone can be higher than the carbon intensity of diesel that it intends to displace for lowering greenhouse gas emissions. This letter focuses narrowly on GHG emissions and does not address socio-economic-ecological prospects for these degraded lands for palm oil or for other purposes. Future studies should carefully evaluate these tradeoffs.

Continuous, real-time bioimaging of chemical bioavailability and toxicology using autonomously bioluminescent human cell lines

NASA Astrophysics Data System (ADS)

Xu, Tingting; Close, Dan M.; Webb, James D.; Price, Sarah L.; Ripp, Steven A.; Sayler, Gary S.

2013-05-01

Bioluminescent imaging is an emerging biomedical surveillance strategy that uses external cameras to detect in vivo light generated in small animal models of human physiology or in vitro light generated in tissue culture or tissue scaffold mimics of human anatomy. The most widely utilized of reporters is the firefly luciferase (luc) gene; however, it generates light only upon addition of a chemical substrate, thus only generating intermittent single time point data snapshots. To overcome this disadvantage, we have demonstrated substrate-independent bioluminescent imaging using an optimized bacterial bioluminescence (lux) system. The lux reporter produces bioluminescence autonomously using components found naturally within the cell, thereby allowing imaging to occur continuously and in real-time over the lifetime of the host. We have validated this technology in human cells with demonstrated chemical toxicological profiling against exotoxin exposures at signal strengths comparable to existing luc systems (~1.33 × 107 photons/second). As a proof-in-principle demonstration, we have engineered breast carcinoma cells to express bioluminescence for real-time screening of endocrine disrupting chemicals and validated detection of 17β-estradiol (EC50 = ~ 10 pM). These and other applications of this new reporter technology will be discussed as potential new pathways towards improved models of target chemical bioavailability, toxicology, efficacy, and human safety.

Bioluminescent imaging demonstrates transplanted human embryonic stem cell-derived CD34+ cells preferentially develop into endothelial cells

PubMed Central

Tian, Xinghui; Hexum, Melinda K.; Penchev, Vesselin R.; Taylor, Russell J.; Shultz, Leonard D.; Kaufman, Dan S

2010-01-01

Human embryonic stem cells (hESCs) provide an important resource for novel regenerative medicine therapies and have been used to derive diverse cell populations, including hematopoietic and endothelial cells. However, it remains a challenge to achieve significant engraftment of hESC-derived blood cells when transplanted into animal models. To better understand mechanisms that enhance or limit the in vivo developmental potential of hESC-derived cells, we utilized hESCs that express firefly luciferase (luc) to allow non-invasive, real-time bioluminescent imaging of hESC-derived CD34+ cells transplanted into the liver of neonatal immunodeficient mice. Serial imaging demonstrated stable engraftment and expansion of the luc+ hESC-derived cells in vivo over several months. While we found that these hESC-derived CD34+ cells have bipotential ability to generate both hematopoietic and endothelial lineages in vitro, these studies demonstrate preferential differentiation into endothelial cells in vivo, with only low levels of hematopoietic cell engraftment. Therefore, these studies reveal key differences in the developmental potential of hESC-derived cells using in vitro and in vivo analyses. While transplanted hESC-derived CD34+ cells are well suited for revascularization therapies, additional measures are needed to provide higher levels of long-term hematopoietic engraftment. PMID:19711457

Land use and land cover changes in Zêzere watershed (Portugal)--Water quality implications.

PubMed

Meneses, B M; Reis, R; Vale, M J; Saraiva, R

2015-09-15

To understand the relations between land use allocation and water quality preservation within a watershed is essential to assure sustainable development. The land use and land cover (LUC) within Zêzere River watershed registered relevant changes in the last decades. These land use and land cover changes (LUCCs) have impacts in water quality, mainly in surface water degradation caused by surface runoff from artificial and agricultural areas, forest fires and burnt areas, and caused by sewage discharges from agroindustry and urban sprawl. In this context, the impact of LUCCs in the quality of surface water of the Zêzere watershed is evaluated, considering the changes for different types of LUC and establishing their possible correlations to the most relevant water quality changes. The results indicate that the loss of coniferous forest and the increase of transitional woodland-shrub are related to increased water's pH; while the growth in artificial surfaces and pastures leads mainly to the increase of soluble salts and fecal coliform concentration. These particular findings within the Zêzere watershed, show the relevance of addressing water quality impact driven from land use and should therefore be taken into account within the planning process in order to prevent water stress, namely within watersheds integrating drinking water catchments. Copyright © 2015 Elsevier B.V. All rights reserved.

Gene silencing in primary and metastatic tumors by small interfering RNA delivery in mice: quantitative analysis using melanoma cells expressing firefly and sea pansy luciferases.

PubMed

Takahashi, Yuki; Nishikawa, Makiya; Kobayashi, Naoki; Takakura, Yoshinobu

2005-07-20

Silencing of oncogenes or other genes contributing to tumor malignancy or progression by RNA interference (RNAi) offers a promising approach to treating tumor patients. To achieve RNAi-based tumor therapy, a small interfering RNA (siRNA) or siRNA-expressing vector needs to be delivered to tumor cells, but little information about its in vivo delivery has been reported. In this study, we examined whether the expression of the target gene in tumor cells can be suppressed by the delivery of RNAi effectors to primary and metastatic tumor cells. To quantitatively evaluate the RNAi effects in tumor cells, mouse melanoma B16-BL6 cells were stably transfected with both firefly (a model target gene) and sea pansy (an internal standard gene) luciferase genes to obtain B16-BL6/dual Luc cells. The target gene expression in subcutaneous primary tumors of B16-BL6/dual Luc cells was significantly suppressed by direct injection of the RNAi effectors followed by electroporation. The expression in metastatic hepatic tumors was also significantly reduced by an intravenous injection of either RNAi effector by the hydrodynamics-based procedure. These results indicate that the both RNAi effectors have a potential to silence target gene in tumor cells in vivo when successfully delivered to tumor cells.

Hepa1-6-FLuc cell line with the stable expression of firefly luciferase retains its primary properties with promising bioluminescence imaging ability.

PubMed

Li, Yasha; Liu, Mengnan; Cui, Jiejie; Yang, Ke; Zhao, Li; Gong, Mengjia; Wang, Yi; He, Yun; He, Tongchuan; Bi, Yang

2018-05-01

Reliable animal models are required for the in vivo study of the molecular mechanisms and effects of chemotherapeutic drugs in hepatocarcinoma. In vivo tracing techniques based on firefly luciferase (FLuc) may optimize the non-invasive monitoring of experimental animals. The present study established a murine Hepa1-6-FLuc cell line that stably expressed a retrovirus-delivered FLuc protein gene. The cell morphology, proliferation, migration and invasion ability of Hepa1-6-FLuc cells were the same as that of the Hepa1-6 cells, and thus is suitable to replace Hepa1-6 cells in the construction of hepatocarcinoma animal models. No differences in subcutaneous tumor mass and its pathomorphology from implanted Hepa1-6-FLuc cells were observed compared with Hepa1-6 control tumors. Bioluminescence imaging indicated that the Luc signal of the Hepa1-6-FLuc cells was consistently strengthened with increases in tumor mass; however, the Luc signal of Hepa1-6-AdFLuc became weaker and eventually disappeared during tumor development. Therefore, compared with the transient expression by adenovirus, stable expression of the FLuc gene in Hepa1-6 cells may better reflect cell proliferation and survival in vivo , and provide a reliable source for the establishment of hepatocarcinoma models.

Input-driven versus turnover-driven controls of simulated changes in soil carbon due to land-use change

NASA Astrophysics Data System (ADS)

Nyawira, S. S.; Nabel, J. E. M. S.; Brovkin, V.; Pongratz, J.

2017-08-01

Historical changes in soil carbon associated with land-use change (LUC) result mainly from the changes in the quantity of litter inputs to the soil and the turnover of carbon in soils. We use a factor separation technique to assess how the input-driven and turnover-driven controls, as well as their synergies, have contributed to historical changes in soil carbon associated with LUC. We apply this approach to equilibrium simulations of present-day and pre-industrial land use performed using the dynamic global vegetation model JSBACH. Our results show that both the input-driven and turnover-driven changes generally contribute to a gain in soil carbon in afforested regions and a loss in deforested regions. However, in regions where grasslands have been converted to croplands, we find an input-driven loss that is partly offset by a turnover-driven gain, which stems from a decrease in the fire-related carbon losses. Omitting land management through crop and wood harvest substantially reduces the global losses through the input-driven changes. Our study thus suggests that the dominating control of soil carbon losses is via the input-driven changes, which are more directly accessible to human management than the turnover-driven ones.

Paranasal sinus mucoceles: our clinical experiments

PubMed Central

Topdag, Murat; Iseri, Mete; Sari, Fatih; Erdogan, Selvet; Keskin, I Gurkan

2015-01-01

Objectives: We present the clinical and radiological features, treatment protocols, and medium-long-term results of our patients following surgery for paranasal sinus mucocele, along with a review of the relevant literature. Materials and methods: A total of 18 patients (11 women and 7 men) who underwent surgery for paranasal sinus mucocele at Kocaeli University Faculty of Medicine, Department of Otolaryngology, between 2006 and 2013 were examined retrospectively. The mean patient age was 41 (range 4-73). Demographic and radiological features, symptoms, treatment protocols, and postoperative outcomes were recorded. Results: The most frequently affected sinus was the maxillary sinus (n=9, 50%) followed by the frontal sinus (n=6, 33%) and sphenoidal sinus (n=3, 16%). The main symptom was headache. Endoscopic marsupialization of the mucocele was applied in all 18 patients, while frontal sinus exploration with the osteoplastic flap procedure was performed in one patient and the Caldwell-Luc operation was performed in another patient. The Caldwell-Luc procedure was subsequently required in one patient (6%) and endoscopic revision surgery was required in another patient (6%). Conclusion: Sinus mucocele that enlarges, eroding the surrounding bone tissue, and induces various clinical symptoms due to the impression of the expansile mass, is treated surgically, and must be planned carefully to prevent serious complications. PMID:26770462

Effects of environmental estrogenic chemicals on AP1 mediated transcription with estrogen receptors alpha and beta.

PubMed

Fujimoto, Nariaki; Honda, Hiroaki; Kitamura, Shigeyuki

2004-01-01

There has been much discussion concerning endocrine disrupting chemicals suspected of exerting adverse effects in both wildlife and humans. Since the majority of these compounds are estrogenic, a large number of in vitro tests for estrogenic characteristics have been developed for screening purpose. One reliable and widely used method is the reporter gene assay employing estrogen receptors (ERs) and a reporter gene with a cis-acting estrogen responsive element (ERE). Other elements such as AP1 also mediate estrogenic signals and the manner of response could be quite different from that of ERE. Since this has yet to be explored, the ER mediated AP1 activity in response to a series of environmental estrogens was investigated in comparison with ERE findings. All the compounds exhibited estrogenic properties with ERE-luc and their AP1 responses were quite similar. These was one exception, however, p,p'-DDT (1,1,1,-trichloro-2,2-bis(p-chlorophenyl)ethane) did not exert any AP1-luc activity, while it appeared to be estrogenic at 10(-7) to 10(-5)M with the ERE action. None of the compounds demonstrated ER beta:AP1 activity. These data suggest that significant differences can occur in responses through the two estrogen pathways depending on environmental chemicals.

Quantifying the uncertainties in life cycle greenhouse gas emissions for UK wheat ethanol

NASA Astrophysics Data System (ADS)

Yan, Xiaoyu; Boies, Adam M.

2013-03-01

Biofuels are increasingly promoted worldwide as a means for reducing greenhouse gas (GHG) emissions from transport. However, current regulatory frameworks and most academic life cycle analyses adopt a deterministic approach in determining the GHG intensities of biofuels and thus ignore the inherent risk associated with biofuel production. This study aims to develop a transparent stochastic method for evaluating UK biofuels that determines both the magnitude and uncertainty of GHG intensity on the basis of current industry practices. Using wheat ethanol as a case study, we show that the GHG intensity could span a range of 40-110 gCO2e MJ-1 when land use change (LUC) emissions and various sources of uncertainty are taken into account, as compared with a regulatory default value of 44 gCO2e MJ-1. This suggests that the current deterministic regulatory framework underestimates wheat ethanol GHG intensity and thus may not be effective in evaluating transport fuels. Uncertainties in determining the GHG intensity of UK wheat ethanol include limitations of available data at a localized scale, and significant scientific uncertainty of parameters such as soil N2O and LUC emissions. Biofuel polices should be robust enough to incorporate the currently irreducible uncertainties and flexible enough to be readily revised when better science is available.

Enzyme and Cancer Cell Selectivity of Nanoparticles: Inhibition of 3D Metastatic Phenotype and Experimental Melanoma by Zinc Oxide.

PubMed

DeLong, Robert K; Mitchell, Jennifer A; Morris, R Tyler; Comer, Jeffrey; Hurst, Miranda N; Ghosh, Kartik; Wanekaya, Adam; Mudge, Miranda; Schaeffer, Ashley; Washington, Laurie L; Risor-Marhanka, Azure; Thomas, Spencer; Marroquin, Shanna; Lekey, Amber; Smith, Joshua J; Garrad, Richard; Aryal, Santosh; Abdelhakiem, Mohamed; Glaspell, Garry P

2017-02-01

Biomedical applications for metal and metal oxide nanoparticles are rapidly increasing. Here their functional impact on two well-characterized model enzymes, Luciferase (Luc) or β-galactosidase (β-Gal) was quantitatively compared. Nickel oxide nanoparticle (NiO-NP) activated β-Gal (>400% control) and boron carbide nanoparticle (B4C-NP) inhibited Luc(<10% control), whereas zinc oxide (ZnO-NP) and cobalt oxide (Co3O4-NP) activated β-Gal to a lesser extent and magnesium oxide (MgO) moderately inhibited both enzymes. Melanoma specific killing was in the order; ZnO > B4C ≥ Cu > MgO > Co3O4 > Fe2O3 > NiO, ZnO-NP inhibiting B16F10 and A375 cells as well as ERK enzyme (>90%) and several other cancer-associated kinases (AKT, CREB, p70S6K). ZnO-NP or nanobelt (NB) serve as photoluminescence (PL) cell labels and inhibit 3-D multi-cellular tumor spheroid (MCTS) growth and were tested in a mouse melanoma model. These results demonstrate nanoparticle and enzyme specific biochemical activity and suggest their utility as new tools to explore the important model metastatic foci 3-D environment and their chemotherapeutic potential.

Relevance of emissions timing in biofuel greenhouse gases and climate impacts.

PubMed

Schwietzke, Stefan; Griffin, W Michael; Matthews, H Scott

2011-10-01

Employing life cycle greenhouse gas (GHG) emissions as a key performance metric in energy and environmental policy may underestimate actual climate change impacts. Emissions released early in the life cycle cause greater cumulative radiative forcing (CRF) over the next decades than later emissions. Some indicate that ignoring emissions timing in traditional biofuel GHG accounting overestimates the effectiveness of policies supporting corn ethanol by 10-90% due to early land use change (LUC) induced GHGs. We use an IPCC climate model to (1) estimate absolute CRF from U.S. corn ethanol and (2) quantify an emissions timing factor (ETF), which is masked in the traditional GHG accounting. In contrast to earlier analyses, ETF is only 2% (5%) over 100 (50) years of impacts. Emissions uncertainty itself (LUC, fuel production period) is 1-2 orders of magnitude higher, which dwarfs the timing effect. From a GHG accounting perspective, emissions timing adds little to our understanding of the climate impacts of biofuels. However, policy makers should recognize that ETF could significantly decrease corn ethanol's probability of meeting the 20% GHG reduction target in the 2007 Energy Independence and Security Act. The added uncertainty of potentially employing more complex emissions metrics is yet to be quantified.

NDEC: A NEA platform for nuclear data testing, verification and benchmarking

NASA Astrophysics Data System (ADS)

Díez, C. J.; Michel-Sendis, F.; Cabellos, O.; Bossant, M.; Soppera, N.

2017-09-01

The selection, testing, verification and benchmarking of evaluated nuclear data consists, in practice, in putting an evaluated file through a number of checking steps where different computational codes verify that the file and the data it contains complies with different requirements. These requirements range from format compliance to good performance in application cases, while at the same time physical constraints and the agreement with experimental data are verified. At NEA, the NDEC (Nuclear Data Evaluation Cycle) platform aims at providing, in a user friendly interface, a thorough diagnose of the quality of a submitted evaluated nuclear data file. Such diagnose is based on the results of different computational codes and routines which carry out the mentioned verifications, tests and checks. NDEC also searches synergies with other existing NEA tools and databases, such as JANIS, DICE or NDaST, including them into its working scheme. Hence, this paper presents NDEC, its current development status and its usage in the JEFF nuclear data project.

AIDS-related health behavior: coping, protection motivation, and previous behavior.

PubMed

Van der Velde, F W; Van der Pligt, J

1991-10-01

The purpose of this study was to examine Rogers' protection motivation theory and aspects of Janis and Mann's conflict theory in the context of AIDS-related health behavior. Subjects were 84 heterosexual men and women and 147 homosexual men with multiple sexual partners; LISREL's path-analysis techniques were used to evaluate the goodness of fit of the structural equation models. Protection motivation theory did fit the data but had considerably more explanatory power for heterosexual than for homosexual subjects (49 vs. 22%, respectively). When coping styles were added, different patterns of findings were found among both groups. Adding variables such as social norms and previous behavior increased the explained variance to 73% for heterosexual subjects and to 44% for homosexual subjects. It was concluded that although protection motivation theory did fit the data fairly adequately, expanding the theory with other variables--especially those related to previous behavior--could improve our understanding of AIDS-related health behavior.

Super-duper chemiluminescent proteins applicable to wide range of bioimaging

NASA Astrophysics Data System (ADS)

Suzuki, Kazushi; Nagai, Takeharu

2017-02-01

We report five new spectral variants of bright luminescent protein made by concatenation of the brightest luciferase, NanoLuc, with various color hues of fluorescent proteins. These proteins, which we call enhanced Nano-lanterns (eNLs), allow five-color live-cell imaging without external light illumination as well as detection of single molecules. Furthermore, eNL-based Ca2+ indicators could be used to image long-term Ca2+ dynamics in iPS-derived cardiomyocytes.

Two-Dimensional, Time-Dependent Plasma Structures of a Hall Effect Thruster

DTIC Science & Technology

2011-09-01

atmospheric pressure to 80 mtorr, is accomplished by a Leybold-Trivac rotary van vacuum pump and the second stage is completed by four 20 in CVI...Thruster”. Physics of Plasmas, 13, 2006. 3. Albarede, Luc, Vanessa Vial, Alexey Lazurenko, Andre Bouchoule, and Michel Dudeck. “Low Frequency Dynamical...Force Research Laboratory Space and Missile Division (AFRL/RZS) 5 Pollux Drive Edwards AFB, CA 93524 DSN 525-5230 AFRL/RZS Approval for public release

The Molecular Epidemiology of Malaria in Western Kenya

DTIC Science & Technology

2002-09-01

including tumor necrosis factor alpha (TNF- α), interleukin-10 (IL-10), transforming growth factor beta (TGF-β), interleukin-6 (IL-6), and interferon gamma...Ricard S, Troesch A, Mallet C, Generenaz L, Evans A, Arveiler D, Luc G, Ruidavets JB, Poirier O. Polymorphisms of the transforming growth factor- beta 1...transforming growth factor- beta 1 and tumour necrosis factor-alpha genes: a technical report. Transpl Immunol 1998 6(3): 193-7. 36. Olomolaiye OO

Advanced Planning Briefing for Industry Proceedings, ’Winning the Information War’ held in Long Branch, New Jersey on May 11 - 12, 1994

DTIC Science & Technology

1994-05-12

Page SESSION IV: "OUT-THINK" THE ENEMY ............ ............... .. 345 Session Overview and Introduction ...... .......... .. 347 Army Global...L C LU 0 cli - 0Dco O)o LUJZ 0 LU 0 LL <O 5;o C)Co W LU 0H 0) MCW < c) 2 <- F«-, C 0 Ir CD cc. 0oz nowo Cl) LUC F- z11. Hz b 0 Z LLIO <C >0 j LL f U

In Vivo Imaging of MDR1A Gene Expression

DTIC Science & Technology

2004-12-01

Engineer PGK-neo and Renilla luciferase cassettes, already available, with appropriate loxP sites, into mdrla locus. Repeat for HSV-tk reporter. The...of the gene-targeting vector. under the control of the PGK promoter. Luc: Renilla luciferase fused in- frame with the translated sequences of exon 2...between the two loxP sites, upstream of the Neo cassette. A cloning strategy was then devised to fuse Renilla luciferase in-frame with the translated

The Role of AhR in Breast Cancer Development

DTIC Science & Technology

2004-07-01

The renilla luciferase vectorphRL-TK (0.05 rtg) was co-transfected with firefly luciferase reporter constructs (0.1 ptg pGudLuc, 0.5-1.0 [tg wildtype...Glo Luciferase system (Promega, Madison, WI ) which allowed sequential reading of the firefly and renilla signals. Cells were lysed according to the...Madison, WI ). The renilla signal was read after quenching the firefly output, thus allowing normalization between sample wells. The normalized firefly

Land Conservation in an Evolving Agricultural Industry: Trade-offs to Consider

NASA Astrophysics Data System (ADS)

Baker, J. S.; Murray, B. C.; McCarl, B. A.; Jackson, R. B.

2008-12-01

This study analyzes the interactions of land conservation policy with biofuel expansion using an economic model of the U.S. forest and agricultural sectors. The world agricultural industry is changing rapidly under emerging market and policy-based pressures. An important driver in the U.S. is the Renewable Fuels Standard (RFS), which mandates significant expansion in biofuels production (up to 36 billion gallons/year by 2022). Traditional land conservation practices such as the Conservation Reserve Program (CRP) are at risk in this changing agricultural climate, as the opportunity costs of reverting to cropland continue to rise. Large- scale reversion of CRP acreage is likely to lead to substantial losses in soil carbon, biodiversity, soil erosion protection, and water quality. However, given the increased competition for land resources, continued efforts to maintain the CRP could induce land use change (LUC) and agricultural development from even more sensitive ecosystems, including native grasslands and forests. This study uses economic modeling to study CRP reversion and LUC under multiple scenarios, including: 1) Baseline assumptions of growth in world agricultural demand and energy prices, with and without CRP reversion; 2) Implementation of the RFS while maintaining the CRP; and 3) RFS with CRP reversion allowed. The study is done using the FASOMGHG model (Lee, McCarl et al, 2008), which is well suited for this analysis as it: 1) Depicts land use competition between crops, pasture, CRP, and forestry over a 100 year period 2) Contains comprehensive GHG accounting across the sectors, 3) Allows land in the CRP to revert to cultivation at an economically optimal rate as land values increase, and 4) Extensively models biofuel and conventional agricultural production possibilities. Results generated to date show significant reversion to cultivation, even under the baseline (36% of the total CRP stock by 2020). Implementing the RFS further pressures conservation

LUC-2-1682 long term maintenance of the Anthony Wayne Suspension Bridge main cables.

DOT National Transportation Integrated Search

2017-01-01

The Anthony Wayne Bridge, Ohio's only suspension bridge, is undergoing an extensive rehabilitation. Prior to taking action to preserve the cables, ODOT must decide what measures to take to evaluate the condition of the cables, how best to rehabilitat...

78 FR 51803 - Culturally Significant Objects Imported for Exhibition Determinations: “Nice: Luc Tuymans”

Federal Register 2010, 2011, 2012, 2013, 2014

2013-08-21

.... 2681, et seq.; 22 U.S.C. 6501 note, et seq.), Delegation of Authority No. 234 of October 1, 1999.... Department of State, SA-5, L/PD, Fifth Floor (Suite 5H03), Washington, DC 20522-0505. Dated: August 14, 2013...

Soil Quality Indexing Strategies for Evaluating Sugarcane Expansion in Brazil

PubMed Central

Cherubin, Maurício R.; Karlen, Douglas L.; Cerri, Carlos E. P.; Franco, André L. C.; Tormena, Cássio A.; Davies, Christian A.; Cerri, Carlos C.

2016-01-01

Increasing demand for biofuel has intensified land-use change (LUC) for sugarcane (Saccharum officinarum) expansion in Brazil. Assessments of soil quality (SQ) response to this LUC are essential for quantifying and monitoring sustainability of sugarcane production over time. Since there is not a universal methodology for assessing SQ, we conducted a field-study at three sites within the largest sugarcane-producing region of Brazil to develop a SQ index (SQI). The most common LUC scenario (i.e., native vegetation to pasture to sugarcane) was evaluated using six SQI strategies with varying complexities. Thirty eight soil indicators were included in the total dataset. Two minimum datasets were selected: one using principal component analysis (7 indicators) and the other based on expert opinion (5 indicators). Non-linear scoring curves were used to interpret the indicator values. Weighted and non-weighted additive methods were used to combine individual indicator scores into an overall SQI. Long-term conversion from native vegetation to extensive pasture significantly decreased overall SQ. In contrast, conversion from pasture to sugarcane had no significant impact on overall SQ at the regional scale, but site-specific responses were found. In general, sugarcane production improved chemical attributes (i.e., higher macronutrient levels and lower soil acidity); however it has negative effects on physical and biological attributes (i.e., higher soil compaction and structural degradation as well as lower soil organic carbon (SOC), abundance and diversity of macrofauna and microbial activity). Overall, we found that simple, user-friendly strategies were as effective as more complex ones for identifying SQ changes. Therefore, as a protocol for SQ assessments in Brazilian sugarcane areas, we recommend using a small number of indicators (e.g., pH, P, K, Visual Evaluation of Soil Structure -VESS scores and SOC concentration) and proportional weighting to reflect chemical

Bystander killing effect of DS-8201a, a novel anti-human epidermal growth factor receptor 2 antibody-drug conjugate, in tumors with human epidermal growth factor receptor 2 heterogeneity.

PubMed

Ogitani, Yusuke; Hagihara, Katsunobu; Oitate, Masataka; Naito, Hiroyuki; Agatsuma, Toshinori

2016-07-01

Antibody-drug conjugates deliver anticancer agents selectively and efficiently to tumor tissue and have significant antitumor efficacy with a wide therapeutic window. DS-8201a is a human epidermal growth factor receptor 2 (HER2)-targeting antibody-drug conjugate prepared using a novel linker-payload system with a potent topoisomerase I inhibitor, exatecan derivative (DX-8951 derivative, DXd). It was effective against trastuzumab emtansine (T-DM1)-insensitive patient-derived xenograft models with both high and low HER2 expression. In this study, the bystander killing effect of DS-8201a was evaluated and compared with that of T-DM1. We confirmed that the payload of DS-8201a, DXd (1), was highly membrane-permeable whereas that of T-DM1, Lys-SMCC-DM1, had a low level of permeability. Under a coculture condition of HER2-positive KPL-4 cells and negative MDA-MB-468 cells in vitro, DS-8201a killed both cells, whereas T-DM1 and an antibody-drug conjugate with a low permeable payload, anti-HER2-DXd (2), did not. In vivo evaluation was carried out using mice inoculated with a mixture of HER2-positive NCI-N87 cells and HER2-negative MDA-MB-468-Luc cells by using an in vivo imaging system. In vivo, DS-8201a reduced the luciferase signal of the mice, indicating suppression of the MDA-MB-468-Luc population; however, T-DM1 and anti-HER2-DXd (2) did not. Furthermore, it was confirmed that DS-8201a was not effective against MDA-MB-468-Luc tumors inoculated at the opposite side of the NCI-N87 tumor, suggesting that the bystander killing effect of DS-8201a is observed only in cells neighboring HER2-positive cells, indicating low concern in terms of systemic toxicity. These results indicated that DS-8201a has a potent bystander effect due to a highly membrane-permeable payload and is beneficial in treating tumors with HER2 heterogeneity that are unresponsive to T-DM1. © 2016 The Authors. Cancer Science published by John Wiley & Sons Australia, Ltd on behalf of Japanese Cancer

Soil Quality Indexing Strategies for Evaluating Sugarcane Expansion in Brazil.

PubMed

Cherubin, Maurício R; Karlen, Douglas L; Cerri, Carlos E P; Franco, André L C; Tormena, Cássio A; Davies, Christian A; Cerri, Carlos C

2016-01-01

Increasing demand for biofuel has intensified land-use change (LUC) for sugarcane (Saccharum officinarum) expansion in Brazil. Assessments of soil quality (SQ) response to this LUC are essential for quantifying and monitoring sustainability of sugarcane production over time. Since there is not a universal methodology for assessing SQ, we conducted a field-study at three sites within the largest sugarcane-producing region of Brazil to develop a SQ index (SQI). The most common LUC scenario (i.e., native vegetation to pasture to sugarcane) was evaluated using six SQI strategies with varying complexities. Thirty eight soil indicators were included in the total dataset. Two minimum datasets were selected: one using principal component analysis (7 indicators) and the other based on expert opinion (5 indicators). Non-linear scoring curves were used to interpret the indicator values. Weighted and non-weighted additive methods were used to combine individual indicator scores into an overall SQI. Long-term conversion from native vegetation to extensive pasture significantly decreased overall SQ. In contrast, conversion from pasture to sugarcane had no significant impact on overall SQ at the regional scale, but site-specific responses were found. In general, sugarcane production improved chemical attributes (i.e., higher macronutrient levels and lower soil acidity); however it has negative effects on physical and biological attributes (i.e., higher soil compaction and structural degradation as well as lower soil organic carbon (SOC), abundance and diversity of macrofauna and microbial activity). Overall, we found that simple, user-friendly strategies were as effective as more complex ones for identifying SQ changes. Therefore, as a protocol for SQ assessments in Brazilian sugarcane areas, we recommend using a small number of indicators (e.g., pH, P, K, Visual Evaluation of Soil Structure -VESS scores and SOC concentration) and proportional weighting to reflect chemical

Adoptive Immunotherapy for Epithelial Ovarian Cancer Using T-cells Simultaneously Targeted to Tumor and Tumor-Associated Macrophages

DTIC Science & Technology

2013-12-01

SCID Beige mice. The clinically useful PET tracer 18fluorodeoxyglucose ( FDG ) was used for this study since FDG is preferentially taken up by tumors with...increased glycolytic activity. However, Figure 9 demonstrates that SKOV-luc tumors are not FDG avid, precluding the further development of this...cells   exhibited   a   selective   increase   in   tracer  uptake  in  the  outer  rim  surrounding  the  tumor.  (B

Tamoxifen Dependent Interaction Between the Estrogen Receptor and a Novel P21 Activated Kinase

DTIC Science & Technology

2002-06-01

binding domain at the C Cv1 cells were cotransfected with ARE4-Luc reporter (100 teric G prti Tse bindin dominat the C ng), pRL-SV40 ( Renilla control...48 h and assayed for luciferase and Renilla Three of the four residues defining a putative het- activity. erotrimeric G protein binding motif were...nases. Taken together, these results demonstrated expression of the control Renilla reporter regulated by that the PAK6 CRIB domain was functional with re

Regulation of AR and (beta)-Catenin Signaling by Pin 1 in Prostate Cancer

DTIC Science & Technology

2006-10-01

hrs. cells were harvested for Luciferase reporter Assay. C, CV-1 cells were transfected with Renilla (2.5ng), G5-Luc (10ng), pBIND-LBD (50ng), pACT-AR...stimulation, cells were cultured in 5 to 10% CDS-FBS. Firefly and internal control Renilla lucif- erase activities were determined using a dual luciferase...reporter assay kit (Pro- mega, Madison, WI), and Renilla activities were not consistently affected by any of the cotransfected vectors. The firefly

In Vivo Imaging of mdrla Gene Expression

DTIC Science & Technology

2005-06-01

svImJ mouse strain, compatible with the ES cells used in our Transgenic Mouse Facility. b. Engineer PGK-neo and Renilla luciferase cassettes...inserted between the two loxP sites, upstream of the Neo cassette. A cloning strategy was then devised to fuse Renilla luciferase in-frame with the...sites: B, BamHI; E, EcoRI; S, ScaI. PGK-neo: neo under the control of the PGK promoter. Luc: Renilla luciferase fused in- frame with the translated

Test Area C-80 Complex Final Range Environmental Assessment, Revision 1

DTIC Science & Technology

2009-08-14

C-80 Complex include gaseous chemical materials from current use of ordnance, smokes, and flares, as well as depleted uranium on TA C-80B from...Smoke grenades (various) 430 Flares (various) 430 OS-4 smoke pot 430 C-80B 466 Fog oil 1,000 gallons Ictus nickel foreign weapon exploitation 20...2007b DU = depleted uranium ; ERP = Environmental Restoration Program; LUC = land use control; POI = point of interest A ffected E nvironm ent C hem

The 52nd International Conference on Medicinal Chemistry (RICT 2016) of the French Medicinal Chemistry Society (SCT) Held in Caen (Normandy).

PubMed

Sapi, Janos; Van Hijfte, Luc; Dallemagne, Patrick

2017-06-21

Outstanding Medchem in France: Guest editors Janos Sapi, Luc Van Hjfte, and Patrick Dallemagne look back at the 52 nd International Conference on Medicinal Chemistry (RICT 2016) held in Caen, France. They discuss the history of the French Medicinal Chemistry Society (Société de Chimie Thérapeutique, SCT) and provide highlights of last year's events, including some key presentations now collected in this Special Issue. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

Genome-Wide Analysis of Translational Control in Tuberous Sclerosis Complex

DTIC Science & Technology

2012-07-01

particular non-AUG codons in the 5’UTR. However, these data was “noisy” and required a machine-learning algorithm to identify TIS codons. We develop...To investigate how nutrient signaling affects the folding of nascent chains, we used firefly luciferase (Luc) as a reporter because of its high...folding as the structural basis for the rapid de novo folding of firefly luciferase. Nat Struct Biol 6(7):697-705. 12. Gupta R, Kasturi P, Bracher A

Entropy Viscosity and L1-based Approximations of PDEs: Exploiting Sparsity

DTIC Science & Technology

2015-10-23

AFRL-AFOSR-VA-TR-2015-0337 Entropy Viscosity and L1-based Approximations of PDEs: Exploiting Sparsity Jean-Luc Guermond TEXAS A & M UNIVERSITY 750...REPORT DATE (DD-MM-YYYY) 09-05-2015 2. REPORT TYPE Final report 3. DATES COVERED (From - To) 01-07-2012 - 30-06-2015 4. TITLE AND SUBTITLE Entropy ...conservation equations can be stabilized by using the so-called entropy viscosity method and we proposed to to investigate this new technique. We

Basic Microsurgery Training Using the Laboratory Rat (Rattus norvegicus)

DTIC Science & Technology

2017-03-23

CHANGES: Have there been any personnel/staffing changes (PI/CI/ AI /TC/lnstructor) since the last IACUC approval of protocol, or annual review? _X_ Yes No If...Include Name, Protocol function - PI/CI/ AI /TC/lnstructor, IACUC approval -Yes/No) Maj Ryan Diepenbrock (PI) IACUC Approval Yes 18Jun 15 DELETIONS...Include Name, Protocol function - PI/CI/ AI /TC/lnstructor, Effective date of deletion) Lt Col Jean Luc Niel (PI) June 2015 Col Bradley Turner (Al)July

Carbon Calculator for Land Use Change from Biofuels Production (CCLUB) Users’ Manual and Technical Documentation

DOE Office of Scientific and Technical Information (OSTI.GOV)

Dunn, Jennifer B.; Qin, Zhangcai; Mueller, Steffen

Themore » $$\\underline{C}$$arbon $$\\underline{C}$$alculator for $$\\underline{L}$$and $$\\underline{U}$$se Change from $$\\underline{B}$$iofuels Production (CCLUB) calculates carbon emissions from land use change (LUC) for four different ethanol production pathways including corn grain ethanol and cellulosic ethanol from corn stover, Miscanthus, and switchgrass. This document discusses the version of CCLUB released September 30, 2014 which includes corn and three cellulosic feedstocks: corn stover, Miscanthus, and switchgrass.« less

A Pyrophosphate Based Energy Generating Module

DTIC Science & Technology

2008-12-01

OMB control number. 1. REPORT DATE DEC 2008 2 . REPORT TYPE N/A 3 . DATES COVERED - 4. TITLE AND SUBTITLE A Pyrophosphate Based Energy...for each substrate (PPi, PEP, and AMP) using double reciprocal Lineweaver-Burk plots of saturation data. 10| 2 10| 3 10|4 10|5 10|6 10|7 10-11 10- 10...Partitioning 1 2 3 4 Bilayer - + + + α-Hemolysin - - + + Upper Reservoir +PPi, +PEP +PPi, +PEP +PPi, +PEP +PPi, +PEP, +Luc Lower Reservoir +AMP

JPRS Report West Europe

DTIC Science & Technology

1988-10-12

in French 8 Jul 88 pp 40-41 [Article by Jacques Gevers: "The Optimistic Mandarin"] [Text] Bourgeois childhood and social fiber, loyalty to Marx and...recently tabled by the govern- ment. Flanked by the two ministers of institutional reforms, Philippe Moureaux and Jean Luc Dehaene, he then presented the...executive and its shift to a proportional system. As for Jean Luch Dehaene, he particularly stressed the need for cooperation among the various levels of

Effect of land use land cover change on soil erosion potential in an agricultural watershed.

PubMed

Sharma, Arabinda; Tiwari, Kamlesh N; Bhadoria, P B S

2011-02-01

Universal soil loss equation (USLE) was used in conjunction with a geographic information system to determine the influence of land use and land cover change (LUCC) on soil erosion potential of a reservoir catchment during the period 1989 to 2004. Results showed that the mean soil erosion potential of the watershed was increased slightly from 12.11 t ha(-1) year(-1) in the year 1989 to 13.21 t ha(-1) year(-1) in the year 2004. Spatial analysis revealed that the disappearance of forest patches from relatively flat areas, increased in wasteland in steep slope, and intensification of cultivation practice in relatively more erosion-prone soil were the main factors contributing toward the increased soil erosion potential of the watershed during the study period. Results indicated that transition of other land use land cover (LUC) categories to cropland was the most detrimental to watershed in terms of soil loss while forest acted as the most effective barrier to soil loss. A p value of 0.5503 obtained for two-tailed paired t test between the mean erosion potential of microwatersheds in 1989 and 2004 also indicated towards a moderate change in soil erosion potential of the watershed over the studied period. This study revealed that the spatial location of LUC parcels with respect to terrain and associated soil properties should be an important consideration in soil erosion assessment process.

PDGF-B Gene Therapy Accelerates Bone Engineering and Oral Implant Osseointegration

PubMed Central

Chang, Po-Chun; Seol, Yang-Jo; Cirelli, Joni A; Pellegrini, Gaia R.; Jin, Qiming; Franco, Lea M.; Goldstein, Steven A.; Chandler, Lois A.; Sosnowski, Barbara; Giannobile, William V.

2009-01-01

Platelet-derived growth factor-BB (PDGF-BB) stimulates repair of healing-impaired chronic wounds such as diabetic ulcers and periodontal lesions. However, limitations in predictability of tissue regeneration occur due in part to transient growth factor bioavailability in vivo. Here, we report that gene delivery of PDGF-B stimulates repair of oral implant extraction socket defects. Alveolar ridge defects were created in rats and were treated at the time of titanium implant installation with a collagen matrix containing an adenoviral (Ad) vector encoding PDGF-B (5.5×108 or 5.5×109 pfu/ml), Ad encoding luciferase (Ad-Luc; 5.5×109 pfu/ml; control) or recombinant human PDGF-BB protein (rhPDGF-BB, 0.3 mg/ml). Bone repair and osseointegration were measured via backscattered SEM, histomorphometry, microcomputed tomography, and biomechanical assessments. Further, a panel of local and systemic safety assessments was performed. Results demonstrated bone repair was accelerated by Ad-PDGF-B and rhPDGF-BB delivery compared to Ad-Luc, with the high dose of Ad-PDGF-B more effective than the low dose. No significant dissemination of the vector construct or alteration of systemic parameters was noted. In summary, gene delivery of Ad-PDGF-B demonstrates regenerative and safety capabilities for bone tissue engineering and osseointegration in alveolar bone defects comparable to rhPDGF-BB protein delivery in vivo. PMID:19741730

Biodegradation, Biocompatibility, and Drug Delivery in Poly(ω-pentadecalactone-co-p-dioxanone) Copolyesters

PubMed Central

Liu, Jie; Jiang, Zhaozhong; Zhang, Shengmin; Liu, Chen; Gross, Richard A.; Kyriakides, Themis R.; Saltzman, W. Mark

2011-01-01

Poly(ω-pentadecalactone-co-p-dioxanone) [poly(PDL-co-DO)] copolyesters are copolymers of an isodimorphic system, which remain semicrystalline over the whole range of compositions. Here, we evaluated enzymatically synthesized poly(PDL-co-DO) copolymers as new materials for biomedical applications. In vivo experiments using mice showed that the copolyesters are well tolerated, with tissue responses that are comparable to poly(p-dioxanone). In addition, the copolymers were found to degrade hydrolytically at controlled rates over a period of several months under physiological conditions. The poly(PDL-co-DO) copolymers with up to 69 mol% DO units were successfully transformed to free-standing nanoparticles that are capable of encapsulating an anticancer drug, doxorubicin, or a polynucleotide, siRNA. Drug- or siRNA-loaded nanoparticles exhibited controlled and continuous release of agent over many weeks. In addition, siLUC-encapsulated poly(PDL-co-DO) nanoparticles were active in inhibiting luciferase gene expression in LUC-RKO cells. Because of substantial differences in structure and hydrophobicity between PDL and DO units, poly(PDL-co-DO) biodegradation rate and physical properties can be tuned over a wide range depending on the copolymer composition. Our results demonstrate that the semicrystalline and biodegradable poly(PDL-co-DO) copolyesters are promising biomaterials to serve as drug carriers, as well as potential raw materials for constructing bioabsorbable sutures and other medical devices. PMID:21641030

High-throughput bioluminescence screening of ubiquitin-proteasome pathway inhibitors from chemical and natural sources.

PubMed

Ausseil, Frederic; Samson, Arnaud; Aussagues, Yannick; Vandenberghe, Isabelle; Creancier, Laurent; Pouny, Isabelle; Kruczynski, Anna; Massiot, Georges; Bailly, Christian

2007-02-01

To discover original inhibitors of the ubiquitin-proteasome pathway, the authors have developed a cell-based bioluminescent assay and used it to screen collections of plant extracts and chemical compounds. They first established a DLD-1 human colon cancer cell line that stably expresses a 4Ubiquitin-Luciferase (4Ub-Luc) reporter protein, efficiently targeted to the ubiquitin-proteasome degradation pathway. The assay was then adapted to 96- and 384-well plate formats and calibrated with reference proteasome inhibitors. Assay robustness was carefully assessed, particularly cell toxicity, and the statistical Z factor value was calculated to 0.83, demonstrating a good performance level of the assay. A total of 18,239 molecules and 15,744 plant extracts and fractions thereof were screened for their capacity to increase the luciferase activity in DLD-1 4Ub-Luc cells, and 21 molecules and 66 extracts inhibiting the ubiquitin-proteasome pathway were identified. The fractionation of an active methanol extract of Physalis angulata L. aerial parts was performed to isolate 2 secosteroids known as physalin B and C. In a cell-based Western blot assay, the ubiquitinated protein accumulation was confirmed after a physalin treatment confirming the accuracy of the screening process. The method reported here thus provides a robust approach to identify novel ubiquitin-proteasome pathway inhibitors in large collections of chemical compounds and natural products.

Hypoxia-inducible vascular endothelial growth factor gene therapy using the oxygen-dependent degradation domain in myocardial ischemia.

PubMed

Kim, Hyun Ah; Lim, Soyeon; Moon, Hyung-Ho; Kim, Sung Wan; Hwang, Ki-Chul; Lee, Minhyung; Kim, Sun Hwa; Choi, Donghoon

2010-10-01

A hypoxia-inducible VEGF expression system with the oxygen-dependent degradation (ODD) domain was constructed and tested to be used in gene therapy for ischemic myocardial disease. Luciferase and VEGF expression vector systems were constructed with or without the ODD domain: pEpo-SV-Luc (or pEpo-SV-VEGF) and pEpo-SV-Luc-ODD (or pEpo-SV-VEGF-ODD). In vitro gene expression efficiency of each vector type was evaluated in HEK 293 cells under both hypoxic and normoxic conditions. The amount of VEGF protein was estimated by ELISA. The VEGF expression vectors with or without the ODD domain were injected into ischemic rat myocardium. Fibrosis, neovascularization, and cardiomyocyte apoptosis were assessed using Masson's trichrome staining, α-smooth muscle actin (α-SMA) immunostaining, and the TUNEL assay, respectively. The plasmid vectors containing ODD significantly improved the expression level of VEGF protein in hypoxic conditions. The enhancement of VEGF protein production was attributed to increased protein stability due to oxygen deficiency. In a rat model of myocardial ischemia, the pEpo-SV-VEGF-ODD group exhibited less myocardial fibrosis, higher microvessel density, and less cardiomyocyte apoptosis compared to the control groups (saline and pEpo-SV-VEGF treatments). An ODD-mediated VEGF expression system that facilitates VEGF-production under hypoxia may be useful in the treatment of ischemic heart disease.

Discovery of N-((4-([1,2,4]triazolo[1,5-a]pyridin-6-yl)-5-(6-methylpyridin-2-yl)-1H-imidazol-2-yl)methyl)-2-fluoroaniline (EW-7197): a highly potent, selective, and orally bioavailable inhibitor of TGF-β type I receptor kinase as cancer immunotherapeutic/antifibrotic agent.

PubMed

Jin, Cheng Hua; Krishnaiah, Maddeboina; Sreenu, Domalapally; Subrahmanyam, Vura B; Rao, Kota S; Lee, Hwa Jeong; Park, So-Jung; Park, Hyun-Ju; Lee, Kiho; Sheen, Yhun Yhong; Kim, Dae-Kee

2014-05-22

A series of 2-substituted-4-([1,2,4]triazolo[1,5-a]pyridin-6-yl)-5-(6-methylpyridin-2-yl)imidazoles was synthesized and evaluated to optimize a prototype inhibitor of TGF-β type I receptor kinase (ALK5), 6. Combination of replacement of a quinoxalin-6-yl moiety of 6 with a [1,2,4]triazolo[1,5-a]pyridin-6-yl moiety, insertion of a methyleneamino linker, and a o-F substituent in the phenyl ring markedly increased ALK5 inhibitory activity, kinase selectivity, and oral bioavailability. The 12b (EW-7197) inhibited ALK5 with IC50 value of 0.013 μM in a kinase assay and with IC50 values of 0.0165 and 0.0121 μM in HaCaT (3TP-luc) stable cells and 4T1 (3TP-luc) stable cells, respectively, in a luciferase assay. Selectivity profiling of 12b using a panel of 320 protein kinases revealed that it is a highly selective ALK5/ALK4 inhibitor. Pharmacokinetic study with 12b·HCl in rats showed an oral bioavailability of 51% with high systemic exposure (AUC) of 1426 ng × h/mL and maximum plasma concentration (Cmax) of 1620 ng/mL. Rational optimization of 6 has led to the identification of a highly potent, selective, and orally bioavailable ALK5 inhibitor 12b.

On-command drug release from nanochains inhibits growth of breast tumors.

PubMed

Peiris, Pubudu M; Tam, Morgan; Vicente, Peter; Abramowski, Aaron; Toy, Randall; Bauer, Lisa; Mayer, Aaron; Pansky, Jenna; Doolittle, Elizabeth; Tucci, Samantha; Schmidt, Erik; Shoup, Christopher; Rao, Swetha; Murray, Kaitlyn; Gopalakrishnan, Ramamurthy; Keri, Ruth A; Basilion, James P; Griswold, Mark A; Karathanasis, Efstathios

2014-06-01

To evaluate the ability of radiofrequency (RF)-triggered drug release from a multicomponent chain-shaped nanoparticle to inhibit the growth of an aggressive breast tumor. A two-step solid phase chemistry was employed to synthesize doxorubicin-loaded nanochains, which were composed of three iron oxide nanospheres and one doxorubicin-loaded liposome assembled in a 100-nm-long linear nanochain. The nanochains were tested in the 4T1-LUC-GFP orthotopic mouse model, which is a highly aggressive breast cancer model. The 4T1-LUC-GFP cell line stably expresses firefly luciferase, which allowed the non-invasive in vivo imaging of tumor response to the treatment using bioluminescence imaging (BLI). Longitudinal BLI imaging showed that a single nanochain treatment followed by application of RF resulted in an at least 100-fold lower BLI signal compared to the groups treated with nanochains (without RF) or free doxorubicin followed by RF. A statistically significant increase in survival time of the nanochain-treated animals followed by RF (64.3 days) was observed when compared to the nanochain-treated group without RF (35.7 days), free doxorubicin-treated group followed by RF (38.5 days), and the untreated group (30.5 days; n=5 animals per group). These studies showed that the combination of RF and nanochains has the potential to effectively treat highly aggressive cancers and prolong survival.

Time trend of clinical cases of Lyme disease in two hospitals in Belgium, 2000-2013.

PubMed

De Keukeleire, Mathilde; Vanwambeke, Sophie O; Kabamba, Benoît; Belkhir, Leila; Pierre, Philippe; Luyasu, Victor; Robert, Annie

2017-12-05

As several studies indicated an increase in Lyme disease (LD), notably in neighbouring countries, concerns have arisen regarding the evolution of Lyme disease in Belgium. In order to confirm or infirm the increase of LD in Belgium, we focused on hospital admissions of patients diagnosed with LD between 2000 and 2013 based on hospital admission databases from two hospitals in Belgium. Hospital databases are a stable recording system. We did a retrospective analysis of the medical files of patients hospitalized with Lyme disease in two Belgian hospitals between 2000 and 2013. The annual number of cases of LD for the two studied Belgian hospitals remained stable between 2000 and 2013, ranging from 1 for the Cliniques universitaires Saint-Luc to 15 for the the Clinique Saint-Pierre. No increasing trend were noted in the estimated annual incidence rate but the average estimated annual incidence rate was higher for the hospital Saint-Pierre (8.1 ± 3.7 per 100,000 inhabitants) than Saint-Luc (2.2 ± 1.5 per 100,000 inhabitants). The number of hospital cases of LD peaked between June and November. Based on hospital admissions with LD, no increasing trend was observed for the period 2000-2013 in the two studied Belgian hospitals. This is in line with other studies carried out in Belgium.

The bZIP transcription factor HY5 interacts with the promoter of the monoterpene synthase gene QH6 in modulating its rhythmic expression.

PubMed

Zhou, Fei; Sun, Tian-Hu; Zhao, Lei; Pan, Xi-Wu; Lu, Shan

2015-01-01

The Artemisia annua L. β-pinene synthase QH6 was previously determined to be circadian-regulated at the transcriptional level, showing a rhythmic fluctuation of steady-state transcript abundances. Here we isolated both the genomic sequence and upstream promoter region of QH6. Different regulatory elements, such as G-box (TGACACGTGGCA, -421 bp from the translation initiation site) which might have effects on rhythmic gene expression, were found. Using the yeast one-hybrid and electrophoretic mobility shift assay (EMSA), we confirmed that the bZIP transcription factor HY5 binds to this motif of QH6. Studies with promoter truncations before and after this motif suggested that this G-box was important for the diurnal fluctuation of the transgenic β-glucuronidase gene (GUS) transcript abundance in Arabidopsis thaliana. GUS gene driven by the promoter region immediately after G-box showed an arrhythmic expression in both light/dark (LD) and constant dark (DD) conditions, whereas the control with G-box retained its fluctuation in both LD and DD. We further transformed A. thaliana with the luciferase gene (LUC) driven by an 1400 bp fragment upstream QH6 with its G-box intact or mutated, respectively. The luciferase activity assay showed that a peak in the early morning disappeared in the mutant. Gene expression analysis also demonstrated that the rhythmic expression of LUC was abolished in the hy5-1 mutant.

3D culture of murine neural stem cells on decellularized mouse brain sections.

PubMed

De Waele, Jorrit; Reekmans, Kristien; Daans, Jasmijn; Goossens, Herman; Berneman, Zwi; Ponsaerts, Peter

2015-02-01

Transplantation of neural stem cells (NSC) in diseased or injured brain tissue is widely studied as a potential treatment for various neurological pathologies. However, effective cell replacement therapy relies on the intrinsic capacity of cellular grafts to overcome hypoxic and/or immunological barriers after transplantation. In this context, it is hypothesized that structural support for grafted NSC will be of utmost importance. With this study, we present a novel decellularization protocol for 1.5 mm thick mouse brain sections, resulting in the generation of acellular three-dimensional (3D) brain sections. Next, the obtained 3D brain sections were seeded with murine NSC expressing both the eGFP and luciferase reporter proteins (NSC-eGFP/Luc). Using real-time bioluminescence imaging, the survival and growth of seeded NSC-eGFP/Luc cells was longitudinally monitored for 1-7 weeks in culture, indicating the ability of the acellular brain sections to support sustained ex vivo growth of NSC. Next, the organization of a 3D maze-like cellular structure was examined using confocal microscopy. Moreover, under mitogenic stimuli (EGF and hFGF-2), most cells in this 3D culture retained their NSC phenotype. Concluding, we here present a novel protocol for decellularization of mouse brain sections, which subsequently support long-term 3D culture of undifferentiated NSC. Copyright © 2014 Elsevier Ltd. All rights reserved.

Generation of recombinant rotaviruses expressing fluorescent proteins using an optimized reverse genetics system.

PubMed

Komoto, Satoshi; Fukuda, Saori; Ide, Tomihiko; Ito, Naoto; Sugiyama, Makoto; Yoshikawa, Tetsushi; Murata, Takayuki; Taniguchi, Koki

2018-04-18

An entirely plasmid-based reverse genetics system for rotaviruses was established very recently. We improved the reverse genetics system to generate recombinant rotavirus by transfecting only 11 cDNA plasmids for its 11 gene segments under the condition of increasing the ratio of the cDNA plasmids for NSP2 and NSP5 genes. Utilizing this highly efficient system, we then engineered infectious recombinant rotaviruses expressing bioluminescent (NanoLuc luciferase) and fluorescent (EGFP and mCherry) reporters. These recombinant rotaviruses expressing reporters remained genetically stable during serial passages. Our reverse genetics approach and recombinant rotaviruses carrying reporter genes will be great additions to the tool kit for studying the molecular virology of rotavirus, and for developing future next-generation vaccines and expression vectors. IMPORTANCE Rotavirus is one of the most important pathogens causing severe gastroenteritis in young children worldwide. In this paper, we describe a robust and simple reverse genetics system based on only rotavirus cDNAs, and its application for engineering infectious recombinant rotaviruses harboring bioluminescent (NanoLuc) and fluorescent (EGFP and mCherry) protein genes. This highly efficient reverse genetics system and recombinant RVAs expressing reporters could be powerful tools for the study of different aspects of rotavirus replication. Furthermore, they may be useful for next-generation vaccine production for this medically important virus. Copyright © 2018 American Society for Microbiology.

PEGylation of polypropylenimine dendrimers: effects on cytotoxicity, DNA condensation, gene delivery and expression in cancer cells.

PubMed

Somani, Sukrut; Laskar, Partha; Altwaijry, Najla; Kewcharoenvong, Paphitchaya; Irving, Craig; Robb, Gillian; Pickard, Benjamin S; Dufès, Christine

2018-06-20

Diaminobutyric polypropylenimine (DAB) dendrimers have been shown to be highly efficient non-viral gene delivery systems for cancer therapy. However, their cytotoxicity currently limits their applications. To overcome this issue, PEGylation of DAB dendrimer, using various PEG molecular weights and dendrimer generations, has been attempted to decrease the cytotoxicity and enhance the DNA condensation, size and zeta potential, cellular uptake and transfection efficacy of these dendriplexes. Among all the PEGylated dendrimers synthesized, generation 3- and generation 4-DAB conjugated to low molecular weight PEG (2 kDa) at a dendrimer: DNA ratio of 20:1 and 10:1 resulted in an increase in gene expression on almost all tested cancer cells lines (by up to 3.2-fold compared to unmodified dendrimer in A431 cells). The highest level of β-galactosidase gene expression (10.07 × 10 -3  ± 0.09 × 10 -3  U/mL) was obtained following treatment of B16F10-Luc cells with G4-dendrimer PEGylated with PEG2K at a dendrimer: DNA ratio of 20:1. These delivery systems significantly decreased cytotoxicity on B16F10-Luc cells, by more than 3.4-fold compared to unmodified dendrimer. PEGylated generations 3- and 4-DAB dendrimers are therefore promising gene delivery systems for cancer therapy, combining low cytotoxicity and high transfection efficacy.

Overcoming endosomal barrier by amphotericin B-loaded dual pH-responsive PDMA-b-PDPA micelleplexes for siRNA delivery.

PubMed

Yu, Haijun; Zou, Yonglong; Wang, Yiguang; Huang, Xiaonan; Huang, Gang; Sumer, Baran D; Boothman, David A; Gao, Jinming

2011-11-22

The endosomal barrier is a major bottleneck for the effective intracellular delivery of siRNA by nonviral nanocarriers. Here, we report a novel amphotericin B (AmB)-loaded, dual pH-responsive micelleplex platform for siRNA delivery. Micelles were self-assembled from poly(2-(dimethylamino)ethyl methacrylate)-block-poly(2-(diisopropylamino)ethyl methacrylate) (PDMA-b-PDPA) diblock copolymers. At pH 7.4, AmB was loaded into the hydrophobic PDPA core, and siRNA was complexed with a positively charged PDMA shell to form the micelleplexes. After cellular uptake, the PDMA-b-PDPA/siRNA micelleplexes dissociated in early endosomes to release AmB. Live cell imaging studies demonstrated that released AmB significantly increased the ability of siRNA to overcome the endosomal barrier. Transfection studies showed that AmB-loaded micelleplexes resulted in significant increase in luciferase (Luc) knockdown efficiency over the AmB-free control. The enhanced Luc knockdown efficiency was abolished by bafilomycin A1, a vacuolar ATPase inhibitor that inhibits the acidification of the endocytic organelles. These data support the central hypothesis that membrane poration by AmB and increased endosomal swelling and membrane tension by a "proton sponge" polymer provided a synergistic strategy to disrupt endosomes for improved intracellular delivery of siRNA. © 2011 American Chemical Society

Suppressed cellular oscillations in after-hours mutant mice are associated with enhanced circadian phase-resetting

PubMed Central

Guilding, Clare; Scott, Fiona; Bechtold, David A; Brown, Timothy M; Wegner, Sven; Piggins, Hugh D

2013-01-01

Within the core molecular clock, protein phosphorylation and degradation play a vital role in determining circadian period. The ‘after-hours’ (Afh) mutation in mouse slows the degradation of the core clock protein Cryptochrome, lengthening the period of the molecular clock in the suprachiasmatic nuclei (SCN) and behavioural wheel-running rhythms. However, we do not yet know how the Afh mutation affects other aspects of physiology or the activity of circadian oscillators in other brain regions. Here we report that daily rhythms of metabolism and ingestive behaviours are altered in these animals, as are PERIOD2::LUCIFERASE (PER2::LUC) rhythms in mediobasal hypothalamic nuclei, which influence these behaviours. Overall there is a trend towards period lengthening and a decrease in amplitude of PER2::LUC rhythms throughout the brain. Imaging of single cells from the arcuate and dorsomedial hypothalamic nuclei revealed this reduction in tissue oscillator amplitude to be due to a decrease in the amplitude, rather than a desynchrony, of single cells. Consistent with existing models of oscillator function, this cellular phenotype was associated with a greater susceptibility to phase-shifting stimuli in vivo and in vitro, with light evoking high-amplitude Type 0 resetting in Afh mutant mice. Together, these findings reveal unexpected consequences of the Afh mutation on the amplitude and synchrony of individual cellular oscillators in the SCN. PMID:23207594

Application of the novel bioluminescent ligand-receptor binding assay to relaxin-RXFP1 system for interaction studies.

PubMed

Wu, Qing-Ping; Zhang, Lei; Shao, Xiao-Xia; Wang, Jia-Hui; Gao, Yu; Xu, Zeng-Guang; Liu, Ya-Li; Guo, Zhan-Yun

2016-04-01

Relaxin is a prototype of the relaxin family peptide hormones and plays important biological functions by binding and activating the G protein-coupled receptor RXFP1. To study their interactions, in the present work, we applied the newly developed bioluminescent ligand-receptor binding assay to the relaxin-RXFP1 system. First, a fully active easily labeled relaxin, in which three Lys residues of human relaxin-2 were replaced by Arg, was prepared through overexpression of a single-chain precursor in Pichia pastoris and in vitro enzymatic maturation. Thereafter, the B-chain N-terminus of the easily labeled relaxin was chemically cross-linked with a C-terminal cysteine residue of an engineered NanoLuc through a disulfide linkage. Receptor-binding assays demonstrated that the NanoLuc-conjugated relaxin retained high binding affinity with the receptor RXFP1 (K d = 1.11 ± 0.08 nM, n = 3) and was able to sensitively monitor binding of a variety of ligands with RXFP1. Using the novel bioluminescent binding assay, we demonstrated that three highly conserved B-chain Arg residues of relaxin-3 had distinct contributions to binding of the receptor RXFP1. In summary, our present work provides a novel bioluminescent ligand-receptor binding assay for the relaxin-RXFP1 system to facilitate their interaction studies, such as characterization of relaxin analogues or screening novel agonists or antagonists of RXFP1.

A method to rapidly and accurately compare relative efficacies of non-invasive imaging reporter genes in a mouse model, and its application to luciferase reporters

PubMed Central

Gil, Jose S.; Machado, Hidevaldo B.; Herschman, Harvey R.

2013-01-01

Purpose Our goal is to develop a simple, quantitative, robust method to compare the efficacy of imaging reporter genes in culture and in vivo. We describe an adenoviral vector-liver transduction procedure, and compare the luciferase reporter efficacies. Procedures Alternative reporter genes are expressed in a common adenoviral vector. Vector amounts used in vivo are based on cell culture titrations, ensuring the same transduction efficacy is used for each vector. After imaging, in vivo and in vitro values are normalized to hepatic vector transduction using quantitative real-time PCR. Results We assayed standard firefly luciferase (FLuc), enhanced firefly luciferase (EFLuc), luciferase 2 (Luc2), humanized Renilla luciferase (hRLuc), Renilla luciferase 8.6-535 (RLuc8.6), and a membrane-bound Gaussia luciferase variant (extGLuc) in cell culture and in vivo. We observed a greater that 100-fold increase in bioluminescent signal for both EFLuc and Luc2 when compared to FLuc, and a greater than 106-fold increase for RLuc8.6 when compared to hRLuc. ExtGLuc was not detectable in liver. Conclusions Our findings contrast, in some cases, with conclusions drawn in prior comparisons of these reporter genes, and demonstrate the need for a standardized method to evaluate alternative reporter genes in vivo. Our procedure can be adapted for reporter genes that utilize alternative imaging modalities (fluorescence, bioluminescence, MRI, SPECT, PET). PMID:21850545

IL-15 Deficient Tax Mice Reveal a Role for IL-1α in Tumor Immunity

PubMed Central

Rauch, Daniel A.; Harding, John C.; Ratner, Lee

2014-01-01

IL-15 is recognized as a promising candidate for tumor immunotherapy and has been described as both a promoter of cancer and a promoter of anti-cancer immunity. IL-15 was discovered in cells transformed by HTLV-1, the etiologic agent of adult T cell leukemia/lymphoma (ATL) and the human retrovirus that carries the Tax oncogene. We have developed the TAX-LUC mouse model of ATL in which Tax expression drives both malignant transformation and luciferase expression, enabling non-invasive imaging of tumorigenesis in real time. To identify the role of IL-15 in spontaneous development of lymphoma in vivo, an IL-15−/− TAX-LUC strain was developed and examined. The absence of IL-15 resulted in aggressive tumor growth and accelerated mortality and demonstrated that IL-15 was not required for Tax-mediated lymphoma but was essential for anti-tumor immunity. Further analysis revealed a unique transcriptional profile in tumor cells that arise in the absence of IL-15 that included a significant increase in the expression of IL-1α and IL-1α-regulated cytokines. Moreover, anti-IL-1α antibodies and an IL-1 receptor antagonist (Anakinra) were used to interrogate the potential of IL-1α targeted therapies in this model. Taken together, these findings identify IL-15 and IL-1α as therapeutic targets in lymphoma. PMID:24416335

Insurgency Organization Case Study: Al-Qaeda in the Maghreb, 2006-Present

DTIC Science & Technology

2008-08-01

Al-Qaeda in Islamic Maghreb: A “ Glocal ” Organization.” Studies in Conflict and Terrorism. 1 June 2008. Mekhennet, Souad; Moss, Michael; Schmitt...Al-Qaeda in Islamic Maghreb: A “ Glocal ” Organization.” Studies in Conflict and Terrorism. 1 June 2008. 21Hansen, Andrew. “al-Qaeda in the Islamic...1 July, 2008. 29 Marret, Jean-Luc. “Al-Qaeda in Islamic Maghreb: A “ Glocal ” Organization.” Studies in Conflict and Terrorism. 1 June 2008. 30

Multimodal Imaging of Pathophysiological Changes and Their Role in Development of Breast Cancer Brain Metastasis

DTIC Science & Technology

2010-09-01

vascular and tissue oxygenation. Moreover, by introducing hypoxia reporter gene ( HRE -luciferase) into breast tumor lines, we will be able to use...luciferase re porter gene under the re gulation of an artificial HIF-1-dependent promoter, 5 HRE (14, 1 5). Integrati on of MRI and BLI will provide...mor hypoxi a. 5 x 10 4 MDA-MB231- HRE -ODD-luc cells were directly inje cted into caudal nucle ar area of right side mouse brain. BLI was applied to

Effect of land use change on soil properties and functions

NASA Astrophysics Data System (ADS)

Tonutare, Tonu; Kõlli, Raimo; Köster, Tiina; Rannik, Kaire; Szajdak, Lech; Shanskiy, Merrit

2014-05-01

For good base of sustainable land management and ecologically sound protection of soils are researches on soil properties and functioning. Ecosystem approach to soil properties and functioning is equally important in both natural and cultivated land use conditions. Comparative analysis of natural and agro-ecosystems formed on similar soil types enables to elucidate principal changes caused by land use change (LUC) and to elaborate the best land use practices for local pedo-ecological conditions. Taken for actual analysis mineral soils' catena - rendzina → brown soils → pseudopodzolic soils → gley-podzols - represent ca 1/3 of total area of Estonian normal mineral soils. All soils of this catena differ substantially each from other by calcareousness, acidity, nutrition conditions, fabric and humus cover type. This catena (representative to Estonian pedo-ecological conditions) starts with drought-prone calcareous soils. Brown (distributed in northern and central Estonia) and pseudopodzolic soils (in southern Estonia) are the most broadly acknowledged for agricultural use medium-textured high-quality automorphic soils. Dispersedly distributed gley-podzols are permanently wet and strongly acid, low-productivity sandy soils. In presentation four complex functions of soils are treated: (1) being a suitable soil environment for plant cover productivity (expressed by annual increment, Mg ha-1 yr-1); (2) forming adequate conditions for decomposition, transformation and conversion of fresh falling litter (characterized by humus cover type); (3) deposition of humus, individual organic compounds, plant nutrition elements, air and water, and (4) forming (bio)chemically variegated active space for soil type specific edaphon. Capacity of soil cover as depositor (3) depends on it thickness, texture, calcareousness and moisture conditions. Biological activity of soil (4) is determined by fresh organic matter influx, quality and quantity of biochemical substances and humus

Land use and land management effects on soil organic carbon stock in Mediterranean agricultural areas (Southern Spain)

NASA Astrophysics Data System (ADS)

Parras-Alcántara, Luis; Lozano-García, Beatriz

2014-05-01

INTRODUCTION Soils play a key role in the carbon geochemical cycle. Agriculture contributes to carbon sequestration through photosynthesis and the incorporation of carbon into carbohydrates. Soil management is one of the best tools for climate change mitigation. Small increases or decreases in soil carbon content due to changes in land use or management practices, may result in a significant net exchange of carbon between the soil carbon pool and the atmosphere. In the last decades arable crops (AC) have been transformed into olive grove cultivations (OG) or vineyards (V) in Mediterranean areas. A field study was conducted to determine long-term effects of land use change (LUC) (AC by OG and V) on soil organic carbon (SOC), total nitrogen (TN), C:N ratio and their stratification in Calcic-Chromic Luvisols (LVcc/cr) in Mediterranean conditions. MATERIAL AND METHODS An unirrigated farm in Montilla-Moriles (Córdoba, Spain) cultivated under conventional tillage (animal power with lightweight reversible plows and non-mineral fertilization or pesticides) was selected for study in 1965. In 1966, the farm was divided into three plots with three different uses (AC, OG and V). The preliminary analyses were realized in 1965 for AC (AC1), and the second analyses were realized in 2011 for AC (AC2 - winter crop rotation with annual wheat and barley, receiving mineral fertilization or pesticides), OG (annual passes with disk harrow and cultivator in the spring, followed by a tine harrow in the summer receiving mineral fertilization and weed control with residual herbicides), and V (with three or five chisel passes a year from early spring to early autumn with mineral fertilization or pesticides.). In all cases (AC1, AC2, OG and V) were collected soil entire profiles. Soil properties determined were: soil particle size, bulk density, SOC, TN, C:N ratio, stocks and SRs. The statistical significance of the differences in the variables between land use practices was tested using the

The K-8 Aeronautics Internet Testbook: Project Plan-Year 3

NASA Technical Reports Server (NTRS)

Pallis, Jani Macari

1997-01-01

The Project Plan - Year 3 includes: the major tasks and milestones, the assignment of tasks, and the key personnel assigned to each task, a description of the milestone and major task, and the deliverables for the project. Milestones have project numbers ending in "0.0", such as "25.0.0 Phase III Development". Major tasks under that milestone begin with the same first number such as, "25.0.1 Develop Future Of Aeronautics". The assignment of tasks included in the Year 3 project plan contains the names of the key personnel responsible for the task and others participating on the task. Although the PI has the overall and ultimately responsible for all the tasks and milestones, the name of the first individual on a task or milestone is the person responsible for that task. Other names listed under that task will be participating on the task. Since there are two individuals at Cislunar Aerospace, Inc. with the name "J. Pallis", the principal investigator, Jani Macari Pallis is designated as PI in the Project Plan and James Pallis is designated as J. Pallis.

Measurement of thermal conductivity of materials down to 4.5 K for development of cryosorption pumps

NASA Astrophysics Data System (ADS)

Verma, Ravi; Behera, Upendra; Kasthurirengan, S.; Shivaprakash, N. C.; Udgata, S. S.; Gangradey, R.

2017-02-01

Cryosorption pumps belong to the class of entrapment or capture vacuum pumps and they retain the gas molecules by sorption and / or by condensation on its internal surfaces. An important aspect in their development is the proper adhesion of the activated carbon granules onto the metallic panel and their cooling to the lowest possible temperature by using high thermal conductivity adhesives for adhering the activated carbons. Hence, the thermal conductivity data of the select adhesives and activated carbons down to 4.5 K are quite essential, but they are not available in open literature. Towards this, an experimental setup has been developed to measure the thermal conductivities of samples with high or low thermal conductivities from 300 K to 4.5 K, with liquid helium using a Janis SuperVariTemp cryostat. This paper presents the details of the experimental setup and the results of our studies on (i) standard samples and (ii) epoxy based adhesives samples. The above studies will enable to make the right choice of adhesives for the development of cryosorption pumps.

Cannabis, alcohol use, psychological distress, and decision-making style.

PubMed

Phillips, James G; Ogeil, Rowan P

2017-09-01

There have been suggestions of hypofrontality in cannabis users. To understand cannabis-related differences in decisional processes, Janis and Mann's conflict model of decision making was applied to recreational cannabis smokers who varied in their alcohol use and level of psychological distress. An online sample of recreational substance users (114 male, 119 female) completed the Melbourne Decision Making Questionnaire, the Alcohol Use Disorders Identification Test (AUDIT), Kessler's Psychological Distress Scale (K10), and the Severity of Dependence Scale (SDS) for cannabis. Multivariate analysis of variance examined self-reported decision-making styles as a function of gender, recent cannabis use, risky alcohol consumption, and levels of psychological distress. Psychological distress was associated with lower decisional self-esteem and higher levels of procrastination and buck-passing. There were gender differences associated with cannabis use. Female cannabis users reported higher levels of hypervigilance, while male cannabis users reported lower levels of buck-passing. Although there was little indication of an avoidant decisional style in cannabis users, the results suggest that cannabis affects decisional processes, contributing to panic in females and impulsivity in males.

Building cosmological frozen stars

NASA Astrophysics Data System (ADS)

Kastor, David; Traschen, Jennie

2017-02-01

Janis-Newman-Winicour (JNW) solutions generalize Schwarzschild to include a massless scalar field. While they share the familiar infinite redshift feature of Schwarzschild, they suffer from the presence of naked singularities. Cosmological versions of JNW spacetimes were discovered some years ago, in the most general case, by Fonarev. Fonarev solutions are also plagued by naked singularities, but have the virtue, unlike e.g. Schwarzschild-deSitter, of being dynamical. Given that exact dynamical cosmological black hole solutions are scarce, Fonarev solutions merit further study. We show how Fonarev solutions can be obtained via generalized dimensional reduction from simpler static vacuum solutions. These results may lead towards constructions of actual dynamical cosmological black holes. In particular, we note that cosmological versions of extremal charged dilaton black holes are known. JNW spacetimes represent a different limiting case of the family of charged dilaton black holes, which have been important in the context of string theory, and better understanding their cosmological versions of JNW spacetimes thus provides a second data point towards finding cosmological versions of the entire family.

The Power of Water

NASA Astrophysics Data System (ADS)

Petrova, Zhaneta; Miteva, Kamelia

2013-04-01

The Power of Water Zh. Petrova, K. Miteva Bio Games, Sofia, Bulgaria (petrova.jani@gmail.com; miteva.kamelia@gmail.com) Lessons "The Power of Water" Due to our belief in the initial creativity of the children and their capacity for discover and perceive logically the world, we consider that the primary and even the pre-school learning have a significant influence in the process of suggesting the idea of respect to the natural forces. These classroom activities include a variety of hand- and self-made simulation models with natural materials and toys which lead the children to easy understanding of what could 'friendly' water do and how powerful, dangerous and not-friendly it could be. During the lessons the children draw their own conclusions of the causes and possible solutions of natural hazards caused by water in each of its forms - avalanches, inundations, floods, the water influence in activation of landslides. The children make on their own some of the models and test them via simulations. In the end they discuss what they have learned in groups.

Saving Energy around the House = Tien Tan Trong Viec Tieu Thu Nang Luc Trong Nha.

ERIC Educational Resources Information Center

Noyes, Marilyn; Jarrett, Von

This bilingual booklet is intended to help Vietnamese refugees learn basic energy conservation skills. Included in the booklet are Vietnamese and English translations of basic energy conservation practices related to the following areas: heating, cooling, cooking, using refrigerators and freezers, lighting, water heating, doing laundry, pursuing…

National Dam Safety Program. Goshen Reservoir Number 1 Dam (Inventory Number N.Y. 488), Lower Hudson River Basin, Orange County, New York. Phase I Inspection Report,

DTIC Science & Technology

1981-08-01

ApprOV~U LUC puoiC release; Distribution unlimited. * ~ . IST~aUTaO STATEMENT (of tho obafact solored In Block 20. It dNagweni hom Ropaot) 146...6 4.1 PROCEDURES 6 4.2 MAINTENANCE OF DAM 6 4.3 WARNING SYSTEM 6 4.4 EVALUATION 6 5 HYDROLOGIC/HYDRAULIC 7 PAGE NO. 5.1 DRAINAGE AREA CHARACTERISTICS...embankment. It houses two 20" intake valves to the Goshen water supply system . The spillway is a concrete channel with a concrete cutoff extending into the

Big River Reservoir Project - Pawcatuck River and Narragansett bay Drainage Basins - Water and related Land Resources Study Volume IV. Attachment I.

DTIC Science & Technology

1981-07-01

U0O00,, 1 lrsLg’ilficant Jlani use changes have I j ixL(LUC L or has5 icure t. Lji river basil invo v- Wv nwj ek Hall ftwu siopping ceti-ers wich1...resources. Pesticides also coul! be diverted with flooe flews and assimilated by fishery resources. As a result of physiological phenomena exhibited by...accumulation relative to quantities of heavy metals and pesticides contained in river flood flows should be presented. Page 4-11, para. 4.60 - We question the

Global Carbon Budget 2016

DOE Office of Scientific and Technical Information (OSTI.GOV)

Le Quéré, Corinne; Andrew, Robbie M.; Canadell, Josep G.

Accurate assessment of anthropogenic carbon dioxide (CO 2) emissions and their redistribution among the atmosphere, ocean, and terrestrial biosphere – the “global carbon budget” – is important to better understand the global carbon cycle, support the development of climate policies, and project future climate change. Here we describe data sets and methodology to quantify all major components of the global carbon budget, including their uncertainties, based on the combination of a range of data, algorithms, statistics, and model estimates and their interpretation by a broad scientific community. We discuss changes compared to previous estimates and consistency within and among components,more » alongside methodology and data limitations. CO 2 emissions from fossil fuels and industry ( E FF) are based on energy statistics and cement production data, respectively, while emissions from land-use change ( E LUC), mainly deforestation, are based on combined evidence from land-cover change data, fire activity associated with deforestation, and models. The global atmospheric CO 2 concentration is measured directly and its rate of growth ( G ATM) is computed from the annual changes in concentration. The mean ocean CO 2 sink ( S OCEAN) is based on observations from the 1990s, while the annual anomalies and trends are estimated with ocean models. The variability in S OCEAN is evaluated with data products based on surveys of ocean CO 2 measurements. The global residual terrestrial CO 2 sink ( S LAND) is estimated by the difference of the other terms of the global carbon budget and compared to results of independent dynamic global vegetation models. We compare the mean land and ocean fluxes and their variability to estimates from three atmospheric inverse methods for three broad latitude bands. All uncertainties are reported as ±1 σ, reflecting the current capacity to characterise the annual estimates of each component of the global carbon budget. For the last decade

Global Carbon Budget 2016

DOE PAGES

Le Quéré, Corinne; Andrew, Robbie M.; Canadell, Josep G.; ...

2016-11-14

Accurate assessment of anthropogenic carbon dioxide (CO 2) emissions and their redistribution among the atmosphere, ocean, and terrestrial biosphere – the “global carbon budget” – is important to better understand the global carbon cycle, support the development of climate policies, and project future climate change. Here we describe data sets and methodology to quantify all major components of the global carbon budget, including their uncertainties, based on the combination of a range of data, algorithms, statistics, and model estimates and their interpretation by a broad scientific community. We discuss changes compared to previous estimates and consistency within and among components,more » alongside methodology and data limitations. CO 2 emissions from fossil fuels and industry ( E FF) are based on energy statistics and cement production data, respectively, while emissions from land-use change ( E LUC), mainly deforestation, are based on combined evidence from land-cover change data, fire activity associated with deforestation, and models. The global atmospheric CO 2 concentration is measured directly and its rate of growth ( G ATM) is computed from the annual changes in concentration. The mean ocean CO 2 sink ( S OCEAN) is based on observations from the 1990s, while the annual anomalies and trends are estimated with ocean models. The variability in S OCEAN is evaluated with data products based on surveys of ocean CO 2 measurements. The global residual terrestrial CO 2 sink ( S LAND) is estimated by the difference of the other terms of the global carbon budget and compared to results of independent dynamic global vegetation models. We compare the mean land and ocean fluxes and their variability to estimates from three atmospheric inverse methods for three broad latitude bands. All uncertainties are reported as ±1 σ, reflecting the current capacity to characterise the annual estimates of each component of the global carbon budget. For the last decade

Global carbon budget 2014

DOE PAGES

Le Quéré, C.; Moriarty, R.; Andrew, R. M.; ...

2015-05-08

Accurate assessment of anthropogenic carbon dioxide (CO 2) emissions and their redistribution among the atmosphere, ocean, and terrestrial biosphere is important to better understand the global carbon cycle, support the development of climate policies, and project future climate change. Here we describe data sets and a methodology to quantify all major components of the global carbon budget, including their uncertainties, based on the combination of a range of data, algorithms, statistics, and model estimates and their interpretation by a broad scientific community. We discuss changes compared to previous estimates, consistency within and among components, alongside methodology and data limitations. COmore » 2 emissions from fossil fuel combustion and cement production ( E FF) are based on energy statistics and cement production data, respectively, while emissions from land-use change ( E LUC), mainly deforestation, are based on combined evidence from land-cover-change data, fire activity associated with deforestation, and models. The global atmospheric CO 2 concentration is measured directly and its rate of growth ( G ATM) is computed from the annual changes in concentration. The mean ocean CO 2 sink ( S OCEAN) is based on observations from the 1990s, while the annual anomalies and trends are estimated with ocean models. The variability in S OCEAN is evaluated with data products based on surveys of ocean CO 2 measurements. The global residual terrestrial CO 2 sink ( S LAND) is estimated by the difference of the other terms of the global carbon budget and compared to results of independent dynamic global vegetation models forced by observed climate, CO 2, and land-cover-change (some including nitrogen–carbon interactions). We compare the mean land and ocean fluxes and their variability to estimates from three atmospheric inverse methods for three broad latitude bands. All uncertainties are reported as ±1σ;, reflecting the current capacity to characterise the

Global Carbon Budget 2015

DOE PAGES

Le Quéré, C.; Moriarty, R.; Andrew, R. M.; ...

2015-12-07

Accurate assessment of anthropogenic carbon dioxide (CO 2) emissions and their redistribution among the atmosphere, ocean, and terrestrial biosphere is important to better understand the global carbon cycle, support the development of climate policies, and project future climate change. Here we describe data sets and a methodology to quantify all major components of the global carbon budget, including their uncertainties, based on the combination of a range of data, algorithms, statistics, and model estimates and their interpretation by a broad scientific community. We also discuss changes compared to previous estimates as well as consistency within and among components, alongside methodologymore » and data limitations. CO 2 emissions from fossil fuels and industry ( E FF) are based on energy statistics and cement production data, while emissions from land-use change ( E LUC), mainly deforestation, are based on combined evidence from land-cover-change data, fire activity associated with deforestation, and models. The global atmospheric CO 2 concentration is measured directly and its rate of growth ( G ATM) is computed from the annual changes in concentration. Moreover, the mean ocean CO 2 sink ( S OCEAN) is based on observations from the 1990s, while the annual anomalies and trends are estimated with ocean models. The variability in S OCEAN is evaluated with data products based on surveys of ocean CO 2 measurements. The global residual terrestrial CO 2 sink ( S LAND) is estimated by the difference of the other terms of the global carbon budget and compared to results of independent dynamic global vegetation models forced by observed climate, CO 2, and land-cover change (some including nitrogen–carbon interactions). We compare the mean land and ocean fluxes and their variability to estimates from three atmospheric inverse methods for three broad latitude bands. All uncertainties are reported as ±1σ, reflecting the current capacity to characterise the annual

Global Carbon Budget 2015

DOE Office of Scientific and Technical Information (OSTI.GOV)

Le Quéré, C.; Moriarty, R.; Andrew, R. M.

Accurate assessment of anthropogenic carbon dioxide (CO 2) emissions and their redistribution among the atmosphere, ocean, and terrestrial biosphere is important to better understand the global carbon cycle, support the development of climate policies, and project future climate change. Here we describe data sets and a methodology to quantify all major components of the global carbon budget, including their uncertainties, based on the combination of a range of data, algorithms, statistics, and model estimates and their interpretation by a broad scientific community. We also discuss changes compared to previous estimates as well as consistency within and among components, alongside methodologymore » and data limitations. CO 2 emissions from fossil fuels and industry ( E FF) are based on energy statistics and cement production data, while emissions from land-use change ( E LUC), mainly deforestation, are based on combined evidence from land-cover-change data, fire activity associated with deforestation, and models. The global atmospheric CO 2 concentration is measured directly and its rate of growth ( G ATM) is computed from the annual changes in concentration. Moreover, the mean ocean CO 2 sink ( S OCEAN) is based on observations from the 1990s, while the annual anomalies and trends are estimated with ocean models. The variability in S OCEAN is evaluated with data products based on surveys of ocean CO 2 measurements. The global residual terrestrial CO 2 sink ( S LAND) is estimated by the difference of the other terms of the global carbon budget and compared to results of independent dynamic global vegetation models forced by observed climate, CO 2, and land-cover change (some including nitrogen–carbon interactions). We compare the mean land and ocean fluxes and their variability to estimates from three atmospheric inverse methods for three broad latitude bands. All uncertainties are reported as ±1σ, reflecting the current capacity to characterise the annual

Global carbon budget 2014

DOE Office of Scientific and Technical Information (OSTI.GOV)

Le Quéré, C.; Moriarty, R.; Andrew, R. M.

Accurate assessment of anthropogenic carbon dioxide (CO 2) emissions and their redistribution among the atmosphere, ocean, and terrestrial biosphere is important to better understand the global carbon cycle, support the development of climate policies, and project future climate change. Here we describe data sets and a methodology to quantify all major components of the global carbon budget, including their uncertainties, based on the combination of a range of data, algorithms, statistics, and model estimates and their interpretation by a broad scientific community. We discuss changes compared to previous estimates, consistency within and among components, alongside methodology and data limitations. COmore » 2 emissions from fossil fuel combustion and cement production ( E FF) are based on energy statistics and cement production data, respectively, while emissions from land-use change ( E LUC), mainly deforestation, are based on combined evidence from land-cover-change data, fire activity associated with deforestation, and models. The global atmospheric CO 2 concentration is measured directly and its rate of growth ( G ATM) is computed from the annual changes in concentration. The mean ocean CO 2 sink ( S OCEAN) is based on observations from the 1990s, while the annual anomalies and trends are estimated with ocean models. The variability in S OCEAN is evaluated with data products based on surveys of ocean CO 2 measurements. The global residual terrestrial CO 2 sink ( S LAND) is estimated by the difference of the other terms of the global carbon budget and compared to results of independent dynamic global vegetation models forced by observed climate, CO 2, and land-cover-change (some including nitrogen–carbon interactions). We compare the mean land and ocean fluxes and their variability to estimates from three atmospheric inverse methods for three broad latitude bands. All uncertainties are reported as ±1σ;, reflecting the current capacity to characterise the

TGF-beta1 stimulates expression of the aromatase (CYP19) gene in human osteoblast-like cells and THP-1 cells.

PubMed

Shozu, M; Zhao, Y; Simpson, E R

2000-02-25

Recent evidence has shown that bone is not only a target of estrogen action but also a source of local estrogen production. Bone cells such as osteoblasts express aromatase (P450arom) and the expression of P450arom in osteoblasts is positively regulated in a tissue specific fashion, as in the case of other tissues which express P450arom. To clarify the physiological factors regulating expression of P450arom in bone, we tested TGF-beta1 using osteoblast-like cells obtained from human fetuses as well as THP-1 cells. TGF-beta1 increased IL-1beta+DEX- induced aromatase activity in osteoblast-like cells, while it inhibited activity in skin fibroblasts. Similar enhancement of aromatase activity by TGF-beta1 was found in DEX-stimulated THP-1 cells and this cell line was used for further experiments. In THP-1 cells, TGF-beta1 enhanced DEX-induced aromatase activity almost linearly by 12 h and thereafter. Increased levels of P450arom transcripts were also demonstrated by RT-PCR at 3 h of TGF-beta1 treatment and thereafter. Cyclohexamide abolished enhancement of activity but did not inhibit the accumulation of P450arom transcripts induced by TGF-beta1. Increase in P450arom expression by TGF-beta1 was attributable to expression driven by promoter I.4. TGF-beta1 did not change the half life of P450arom transcripts. To identify the cis-acting elements responsible for TGF-beta1 action on aromatase expression, transient transfection assays were performed using a series of deletion constructs for promoter I.4 (P450-I.4/Luc). Two constructs (-410/+14 and-340/+14) that contain a functional glucocorticoid response element (GRE) and downstream sequence showed significant increase of luciferase activity in response to TGF-beta1. Deletion and mutation of the GRE in P450-I.4/Luc (-340/+14) abolished the TGF-beta1. The luciferase activity of a (GRE)(1)-SV40/Luc construct was also stimulated by TGF-beta1. These results indicate that TGF-beta1 increases the expression of P450arom at the

Tracking changes in land-use and drainage status of organic soils using heterogeneous spatial datasets

NASA Astrophysics Data System (ADS)

Untenecker, Johanna; Tiemeyer, Bärbel; Freibauer, Annette; Laggner, Andreas; Luterbacher, Jürg

2016-04-01

Tracking land-use since 1990 is one of the major challenges in greenhouse gas (GHG) reporting under the United Nations Framework Convention on Climate Change (UNFCCC) and the Kyoto Protocol, as the data availability, especially for the base year 1990, is often poor. Even if data is available, spatial and thematic resolution will often change over time or differ even within one country. Such inconsistencies will cause a strong overestimation of land use change (LUC) if not adequately accounted for. Using different spatial datasets, we present a method that allows tracking changes in land-use and drainage status of organic soils. The drainage status is relevant for the Kyoto activities grazing land management (GM) and wetland drainage and rewetting (WDR) as the GHG emissions of organic soils strongly depend on the groundwater level. We used datasets that are already used for the German national inventory report (Digital Landscape Model of official cadastre data) and high resolution spatial datasets (CIR aerial photography) derived for biodiversity monitoring of six federal states in North and East Germany. This data is combined with the legal protection status such as nature conservation areas. To create a consistent time series, we developed a translation key which allows quantifying gross and net LUC in a spatially explicit manner. The developed method fills the lack of data for 1990 and allows GHG accounting on higher Tier levels as soon as detailed emission factors are ready to be implemented. LUC can be stratified by the protection status. Areas without a protection status show a trend towards both intensification of land-use and drier conditions. Highly protected areas show an opposite trend while a moderate protection level (e.g. by nature parks) did only have very weak effects. Furthermore, there are major differences between federal states. In Schleswig-Holstein, known as a federal state of high agricultural production, organic soils tend to become drier and

Tumour suppressor protein p53 regulates the stress activated bilirubin oxidase cytochrome P450 2A6

DOE Office of Scientific and Technical Information (OSTI.GOV)

Hu, Hao, E-mail: hao.hu1@uqconnect.edu.au; Yu, Ting, E-mail: t.yu2@uq.edu.au; Arpiainen, Satu, E-mail: Satu.Juhila@orion.fi

2015-11-15

Human cytochrome P450 (CYP) 2A6 enzyme has been proposed to play a role in cellular defence against chemical-induced oxidative stress. The encoding gene is regulated by various stress activated transcription factors. This paper demonstrates that p53 is a novel transcriptional regulator of the gene. Sequence analysis of the CYP2A6 promoter revealed six putative p53 binding sites in a 3 kb proximate promoter region. The site closest to transcription start site (TSS) is highly homologous with the p53 consensus sequence. Transfection with various stepwise deletions of CYP2A6-5′-Luc constructs – down to − 160 bp from the TSS – showed p53 responsivenessmore » in p53 overexpressed C3A cells. However, a further deletion from − 160 to − 74 bp, including the putative p53 binding site, totally abolished the p53 responsiveness. Electrophoretic mobility shift assay with a probe containing the putative binding site showed specific binding of p53. A point mutation at the binding site abolished both the binding and responsiveness of the recombinant gene to p53. Up-regulation of the endogenous p53 with benzo[α]pyrene – a well-known p53 activator – increased the expression of the p53 responsive positive control and the CYP2A6-5′-Luc construct containing the intact p53 binding site but not the mutated CYP2A6-5′-Luc construct. Finally, inducibility of the native CYP2A6 gene by benzo[α]pyrene was demonstrated by dose-dependent increases in CYP2A6 mRNA and protein levels along with increased p53 levels in the nucleus. Collectively, the results indicate that p53 protein is a regulator of the CYP2A6 gene in C3A cells and further support the putative cytoprotective role of CYP2A6. - Highlights: • CYP2A6 is an immediate target gene of p53. • Six putative p53REs located on 3 kb proximate CYP2A6 promoter region. • The region − 160 bp from TSS is highly homologous with the p53 consensus sequence. • P53 specifically bind to the p53RE on the − 160 bp region. �

bmo-miR-275 down-regulates expression of Bombyx mori sericin gene 2 in vitro

PubMed Central

Qian, Ping; Jiang, Tao; Wang, Xin; Song, Fei; Chen, Chen; Shen, Xingjia

2018-01-01

We hypothesized that bmo-miR-275 has a potential regulatory function regarding the expression of sericin gene 2 (BmSer-2). First, we examined the expression of bmo-miR-275 and its target gene BmSer-2 in seven different tissues from 5th instar day-3 silkworm larvae. The results showed that they were both specifically expressed in the middle silk gland, implying that spatio-temporal conditions are required for bmo-miR-275 to regulate the expression of BmSer-2. To test this hypothesis, we constructed a pri-bmo-miR-275 expressing plasmid pcDNA3.0 [ie1-egfp-pri-bmo-miR-275-SV40] and BmSer-2-3´UTR recombinant reporter plasmids pGL3.0 [A3-luc-Ser-2-3′ UTR-SV40]. Finally, BmN cells were harvested and luciferase activity was detected. Results showed that luciferase activity was reduced significantly (P<0.05) in BmN cells co-transfected with pcDNA3.0 [ie1-egfp-pri-bmo-miR-275-SV40] and pGL3.0 [A3-luc-Ser-2-3’UTR-SV40], suggesting that bmo-miR-275 can down-regulate the expression of BmSer-2 in vitro. Our results improve the understanding of the regulatory function of Bombyx mori miRNA on the expression of genes regulating silk formation. PMID:29381729

Fibroblast growth factor 2 regulates bone sialoprotein gene transcription in human breast cancer cells.

PubMed

Li, Zhengyang; Wang, Zhitao; Yang, Li; Li, Xinyue; Sasaki, Yoko; Wang, Shuang; Araki, Shouta; Mezawa, Masaru; Takai, Hideki; Nakayama, Youhei; Ogata, Yorimasa

2010-03-01

Bone sialoprotein (BSP) is a major non-collagenous, extracellular matrix glycoprotein associated with mineralized tissues. Fibroblast growth factor 2 (FGF2) is recognized as a potent mitogen for a variety of mesenchymal cells. FGF2 produced by osteoblasts accumulates in the bone matrix and acts as an autocrine/paracrine regulator of osteoblasts. We previously reported that FGF2 regulates BSP gene transcription through the FGF2 response element (FRE) and activator protein 1 (AP1) binding site overlapping with the glucocorticoid response element in the rat BSP gene promoter. In the present study, FGF2 (10 ng/ml) increased BSP and Runx2 mRNA levels at 6 h in MCF7 human breast cancer cells. Transient transfection analyses were performed using chimeric constructs of the human BSP gene promoter linked to a luciferase reporter gene. Treatment of MCF7 cells with FGF2 (10 ng/ml) increased the luciferase activity of the constructs between -84LUC and -927LUC. Gel mobility shift analyses showed that FGF2 increased the binding of AP1 and CRE2. The CRE2- and AP1-protein complexes were disrupted by antibodies against CREB1, c-Fos, c-Jun, Fra2, p300 and Runx2. These studies demonstrate that FGF2 stimulates BSP transcription in MCF7 human breast cancer cells by targeting the AP1 and CRE2 elements in the human BSP gene promoter.

Greenhouse gas emissions and land use change from Jatropha curcas-based jet fuel in Brazil.

PubMed

Bailis, Robert E; Baka, Jennifer E

2010-11-15

This analysis presents a comparison of life-cycle GHG emissions from synthetic paraffinic kerosene (SPK) produced as jet fuel substitute from jatropha curcas feedstock cultivated in Brazil against a reference scenario of conventional jet fuel. Life cycle inventory data are derived from surveys of actual Jatropha growers and processors. Results indicate that a baseline scenario, which assumes a medium yield of 4 tons of dry fruit per hectare under drip irrigation with existing logistical conditions using energy-based coproduct allocation methodology, and assumes a 20-year plantation lifetime with no direct land use change (dLUC), results in the emissions of 40 kg CO₂e per GJ of fuel produced, a 55% reduction relative to conventional jet fuel. However, dLUC based on observations of land-use transitions leads to widely varying changes in carbon stocks ranging from losses in excess of 50 tons of carbon per hectare when Jatropha is planted in native cerrado woodlands to gains of 10-15 tons of carbon per hectare when Jatropha is planted in former agro-pastoral land. Thus, aggregate emissions vary from a low of 13 kg CO₂e per GJ when Jatropha is planted in former agro-pastoral lands, an 85% decrease from the reference scenario, to 141 kg CO₂e per GJ when Jatropha is planted in cerrado woodlands, a 60% increase over the reference scenario. Additional sensitivities are also explored, including changes in yield, exclusion of irrigation, shortened supply chains, and alternative allocation methodologies.

On-command drug release from nanochains inhibits growth of breast tumors

PubMed Central

Peiris, Pubudu M.; Tam, Morgan; Vicente, Peter; Abramowski, Aaron; Toy, Randall; Bauer, Lisa; Mayer, Aaron; Pansky, Jenna; Doolittle, Elizabeth; Tucci, Samantha; Schmidt, Erik; Shoup, Christopher; Rao, Swetha; Murray, Kaitlyn; Gopalakrishnan, Ramamurthy; Keri, Ruth A.; Basilion, James P.; Griswold, Mark A.; Karathanasis, Efstathios

2013-01-01

Purpose To evaluate the ability of radiofrequency (RF)-triggered drug release from a multicomponent chain-shaped nanoparticle to inhibit the growth of an aggressive breast tumor. Methods A two-step solid phase chemistry was employed to synthesize doxorubicin-loaded nanochains, which were composed of three iron oxide nanospheres and one doxorubicin-loaded liposome assembled in a 100-nm-long linear nanochain. The nanochains were tested in the Luc-GFP-4T1 orthotopic mouse model, which is a highly aggressive breast cancer model. The Luc-GFP-4T1 cell line stably expresses firefly luciferase, which allowed the non-invasive in vivo imaging of tumor response to the treatment using bioluminescence imaging (BLI). Results Longitudinal BLI imaging showed that a single nanochain treatment followed by application of RF resulted in an at least 100-fold lower BLI signal compared to the groups treated with nanochains (without RF) or free doxorubicin followed by RF. A statistically significant increase in survival time of the nanochain-treated animals followed by RF (64.3 days) was observed when compared to the nanochain-treated group without RF (35.7 days), free doxorubicin-treated group followed by RF (38.5 days), and the untreated group (30.5 days; n=5 animals per group). Conclusions These studies showed that the combination of RF and nanochains has the potential to effectively treat highly aggressive cancers and prolong survival. PMID:23934254

Overexpression of miRNA-497 inhibits tumor angiogenesis by targeting VEGFR2

DOE Office of Scientific and Technical Information (OSTI.GOV)

Tu, Yingfeng; Liu, Li; Zhao, Dongliang

Recent studies reported miR-497 exhibited inhibitory effects in various cancers. However, whether miR-497 is involved in inhibiting angiogenesis, which is critical for tumor growth and metastasis, is still unknown. The purpose of this study was to investigate the potential role of miR-497 in tumor angiogenesis. In this work, cell proliferation and apoptosis analyses were conducted to explore the potential function of miR-497 in HUVECs by using MTT and TUNEL assays. Western blotting (WB) was employed to validate the downstream targets of miR-497. Furthermore, in order to disclose the role of miR-497 on angiogenesis, VEGFR2-luc transgenic mice were treated with miR-497more » mimic and applied to monitor tumor angiogenesis and growth by in vivo bioluminescent imaging (BLI). The results demonstrated that overexpression of miR-497 showed inhibitory effects on VEGFR2 activation and downstream Raf/MEK/ERK signal pathways in vitro and in vivo. Moreover, overexpression of miR-497 effectively induced HUVECs apoptosis by targeting VEGFR2 and downstream PI3K/AKT signaling pathway. Furthermore, miR-497 exhibited anti-angiogenesis and anti-tumor effects in the VEGFR2-luc breast tumor model proven by BLI, WB and immunohistochemistry analysis. In summary, miR-497 inhibits tumor angiogenesis and growth via targeting VEGFR2, indicating miR-497 can be explored as a potential drug candidate for cancer therapy.« less

Lithium Impacts on the Amplitude and Period of the Molecular Circadian Clockwork

PubMed Central

Li, Jian; Lu, Wei-Qun; Beesley, Stephen; Loudon, Andrew S. I.; Meng, Qing-Jun

2012-01-01

Lithium salt has been widely used in treatment of Bipolar Disorder, a mental disturbance associated with circadian rhythm disruptions. Lithium mildly but consistently lengthens circadian period of behavioural rhythms in multiple organisms. To systematically address the impacts of lithium on circadian pacemaking and the underlying mechanisms, we measured locomotor activity in mice in vivo following chronic lithium treatment, and also tracked clock protein dynamics (PER2::Luciferase) in vitro in lithium-treated tissue slices/cells. Lithium lengthens period of both the locomotor activity rhythms, as well as the molecular oscillations in the suprachiasmatic nucleus, lung tissues and fibroblast cells. In addition, we also identified significantly elevated PER2::LUC expression and oscillation amplitude in both central and peripheral pacemakers. Elevation of PER2::LUC by lithium was not associated with changes in protein stabilities of PER2, but instead with increased transcription of Per2 gene. Although lithium and GSK3 inhibition showed opposing effects on clock period, they acted in a similar fashion to up-regulate PER2 expression and oscillation amplitude. Collectively, our data have identified a novel amplitude-enhancing effect of lithium on the PER2 protein rhythms in the central and peripheral circadian clockwork, which may involve a GSK3-mediated signalling pathway. These findings may advance our understanding of the therapeutic actions of lithium in Bipolar Disorder or other psychiatric diseases that involve circadian rhythm disruptions. PMID:22428012