Real-Time Characterization of Virulence Factor Expression in Yersinia pestis Using a Green Fluorescent Protein Reporter System

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

Forde, C; Rocco, J; Fitch, J P

2004-06-09

A real-time reporter system was developed to monitor the thermal induction of virulence factors in Yersinia pestis. The reporter system consists of a plasmid in Y. pestis in which the expression of green fluorescent protein (GFP) is under the control of the promoters for six virulence factors, yopE, sycE, yopK, yopT, yscN, and lcrE/yopN, which are all components of the Type III secretion virulence mechanism of Y. pestis. Induction of the expression of these genes in vivo was determined by the increase in fluorescence intensity of GFP in real time. Basal expression levels observed for the Y. pestis promoters, expressedmore » as percentages of the positive control with GFP under the control of the lac promoter, were: yopE (15%), sycE (15%), yopK (13%), yopT (4%), lcrE (3.3%) and yscN (0.8%). The yopE reporter showed the strongest gene induction following temperature transition from 26 C to 37 C. The induction levels of the other virulence factors, expressed as percentages of yopE induction, were: yopK (57%), sycE (9%), yscN (3%), lcrE (3%), and yopT (2%). The thermal induction of each of these promoter fusions was repressed by calcium, and the ratios of the initial rates of thermal induction without calcium supplementation compared to the rate with calcium supplementation were: yopE (11 fold), yscN (7 fold), yopK (6 fold), lcrE (3 fold), yopT (2 fold), and sycE (2 fold). This work demonstrates a novel approach to quantify gene induction and provides a method to rapidly determine the effects of external stimuli on expression of Y. pestis virulence factors in real time, in living cells.« less

Demonstration of plant fluorescence by imaging technique and Intelligent FluoroSensor

NASA Astrophysics Data System (ADS)

Lenk, Sándor; Gádoros, Patrik; Kocsányi, László; Barócsi, Attila

2015-10-01

Photosynthesis is a process that converts carbon-dioxide into organic compounds, especially into sugars, using the energy of sunlight. The absorbed light energy is used mainly for photosynthesis initiated at the reaction centers of chlorophyll-protein complexes, but part of it is lost as heat and chlorophyll fluorescence. Therefore, the measurement of the latter can be used to estimate the photosynthetic activity. The basic method, when illuminating intact leaves with strong light after a dark adaptation of at least 20 minutes resulting in a transient change of fluorescence emission of the fluorophore chlorophyll-a called `Kautsky effect', is demonstrated by an imaging setup. The experimental kit includes a high radiant blue LED and a CCD camera (or a human eye) equipped with a red transmittance filter to detect the changing fluorescence radiation. However, for the measurement of several fluorescence parameters, describing the plant physiological processes in detail, the variation of several excitation light sources and an adequate detection method are needed. Several fluorescence induction protocols (e.g. traditional Kautsky, pulse amplitude modulated and excitation kinetic), are realized in the Intelligent FluoroSensor instrument. Using it, students are able to measure different plant fluorescence induction curves, quantitatively determine characteristic parameters and qualitatively interpret the measured signals.

Fluorescent Arc/Arg3.1 indicator mice: a versatile tool to study brain activity changes in vitro and in vivo

PubMed Central

Grinevich, Valery; Kolleker, Alexander; Eliava, Marina; Takada, Naoki; Takuma, Hiroshi; Fukazawa, Yugo; Shigemoto, Ryuichi; Kuhl, Dietmar; Waters, Jack; Seeburg, Peter H.; Osten, Pavel

2014-01-01

The brain-specific immediate early gene Arc/Arg3.1 is induced in response to a variety of stimuli, including sensory and behavior-linked neural activity. Here we report the generation of transgenic mice, termed TgArc/Arg3.1-d4EGFP, expressing a 4-hour half-life form of enhanced green fluorescent protein (d4EGFP) under the control of the Arc/Arg3.1 promoter. We show that d4EGFP-mediated fluorescence faithfully reports Arc/Arg3.1 induction in response to physiological, pathological and pharmacological stimuli, and that this fluorescence permits electrical recording from activated neurons in the live mouse. Moreover, the fluorescent Arc/Arg3.1 indicator revealed activity changes in circumscribed brain areas in distinct modes of stress and in a mouse model of Alzheimer’s disease. These findings identify the TgArc/Arg3.1-d4EGFP mouse as a versatile tool to monitor Arc/Arg3.1 induction in neural circuits, both in vitro and in vivo. PMID:19628007

A plasmid-based Escherichia coli gene expression system with cell-to-cell variation below the extrinsic noise limit

PubMed Central

2017-01-01

Experiments in synthetic biology and microbiology can benefit from protein expression systems with low cell-to-cell variability (noise) and expression levels precisely tunable across a useful dynamic range. Despite advances in understanding the molecular biology of microbial gene regulation, many experiments employ protein-expression systems exhibiting high noise and nearly all-or-none responses to induction. I present an expression system that incorporates elements known to reduce gene expression noise: negative autoregulation and bicistronic transcription. I show by stochastic simulation that while negative autoregulation can produce a more gradual response to induction, bicistronic expression of a repressor and gene of interest can be necessary to reduce noise below the extrinsic limit. I synthesized a plasmid-based system incorporating these principles and studied its properties in Escherichia coli cells, using flow cytometry and fluorescence microscopy to characterize induction dose-response, induction/repression kinetics and gene expression noise. By varying ribosome binding site strengths, expression levels from 55–10,740 molecules/cell were achieved with noise below the extrinsic limit. Individual strains are inducible across a dynamic range greater than 20-fold. Experimental comparison of different regulatory networks confirmed that bicistronic autoregulation reduces noise, and revealed unexpectedly high noise for a conventional expression system with a constitutively expressed transcriptional repressor. I suggest a hybrid, low-noise expression system to increase the dynamic range. PMID:29084263

Dual-labeling with 5-aminolevulinic acid and fluorescein for fluorescence-guided resection of high-grade gliomas: technical note.

PubMed

Suero Molina, Eric; Wölfer, Johannes; Ewelt, Christian; Ehrhardt, André; Brokinkel, Benjamin; Stummer, Walter

2018-02-01

OBJECTIVE Fluorescence guidance with 5-aminolevulinic acid (5-ALA) helps improve resections of malignant gliomas. However, one limitation is the low intensity of blue light for background illumination. Fluorescein has recently been reintroduced into neurosurgery, and novel microscope systems are available for visualizing this fluorochrome, which highlights all perfused tissues but has limited selectivity for tumor detection. Here, the authors investigate a combination of both fluorochromes: 5-ALA for distinguishing tumor and fluorescein for providing tissue fluorescence of adjacent brain tissue. METHODS The authors evaluated 6 patients who harbored cerebral lesions suggestive of high-grade glioma. Patients received 5-ALA (20 mg/kg) orally 4 hours before induction of anesthesia. Low-dose fluorescein (3 mg/kg intravenous) was injected immediately after anesthesia induction. Pentero microscopes (equipped either with Yellow 560 or Blue 400 filters) were used to visualize fluorescence. To simultaneously visualize both fluorochromes, the Yellow 560 module was combined with external blue light illumination (D-light C System). RESULTS Fluorescein-induced fluorescence created a useful background for protoporphyrin IX (PPIX) fluorescence, which appeared orange to red, surrounded by greenly fluorescent normal brain and edematous tissue. Green brain-tissue fluorescence was helpful in augmenting background. Levels of blue illumination that were too strong obscured PPIX fluorescence. Unspecific extravasation of fluorescein was noted at resection margins, which did not interfere with PPIX fluorescence detection. CONCLUSIONS Dual labeling with both PPIX and fluorescein fluorescence is feasible and gives superior background information during fluorescence-guided resections. The authors believe that this technique carries potential as a next step in fluorescence-guided resections if it is completely integrated into the surgical microscope.

A Novel Yeast Surface Display Method for Large-Scale Screen Inhibitors of Sortase A.

PubMed

Wu, Lin; Li, Huijun; Tang, Tianle

2017-01-24

Fluorescence resonance energy transfer substrates of sortase A are too expensive to be used to roughly screen high-throughput sortase A inhibitors. This makes therapeutic strategies difficult to realize in a clinical therapeutic use. Instead, we design here an LPETG-EGFP (leucine, proline, glutamic, threonine and glycine-enhanced green fluorescence) protein displayed on a yeast surface as a substrate by adaptively reducing the cost. We do this by optimizing the induction conditions of sortase A expression in Escherichia coli DE3(BL21) and catalyzing LPETG proteins, which are displayed on surface of Pichia pastoris . Different expression conditions of sortase A include: induction temperature (22 °C, 28 °C, 37 °C and 40 °C), induction time (4 h, 5 h, 6 h and 7 h) and induction concentration of isopropyl β-d-thiogalactoside IPTG (0.25 mmol/L, 0.5 mmol/L, 1 mmol/L, and 2 mmol/L). The fluorescence change of the LPETG-EGFP protein on the surface of P. pastoris over time was detected by flow cytometry and fluorescence spectrophotometry, and then the sensitivities of the two methods were compared. Using berberine chloride as an inhibitor, the activity of sortase A was investigated with the substrates of LPETG-EGFP protein, and compared to Dabcyl-QALPETGEE-Edans. A high yield of sortase A was achieved by inducing 1.0 mmol/L IPTG at 28 °C for 6 h. The intensity of green fluorescence of substrates displayed on the yeast surface was increased over time, while the stability was decreased slightly. Both fluorescence spectrophotometery and flow cytometry were fit for detection because of their high sensitivity. We utilized two different substrates of sortase A to investigate sortase A activity, which resulted in the increase of fluorescence intensity with respect to the increased time of growth. However, the method with Dabcyl-QALPETGEE-Edans as its substrate was more robust. Thus, the method described in this paper is a simple and cheap method which is very suitable for high-throughput analysis, but the conventional method is much more sensitive. The method described in this paper is expected to lead to large-scale screening of sortase A inhibitors which can be used to decrease the risk of drug resistance development.

Development and Characterization of a Green Fluorescent Protein-Based Bacterial Biosensor for Bioavailable Toluene and Related Compounds†

PubMed Central

Stiner, Lawrence; Halverson, Larry J.

2002-01-01

A green fluorescent protein-based Pseudomonas fluorescens strain A506 biosensor was constructed and characterized for its potential to measure benzene, toluene, ethylbenzene, and related compounds in aqueous solutions. The biosensor is based on a plasmid carrying the toluene-benzene utilization (tbu) pathway transcriptional activator TbuT from Ralstonia pickettii PKO1 and a transcriptional fusion of its promoter PtbuA1 with a promoterless gfp gene on a broad-host-range promoter probe vector. TbuT was not limiting, since it was constitutively expressed by being fused to the neomycin phosphotransferase (nptII) promoter. The biosensor cells were readily induced, and fluorescence emission after induction periods of 3 h correlated well with toluene, benzene, ethylbenzene, and trichloroethylene concentrations. Our experiments using flow cytometry show that intermediate levels of gfp expression in response to toluene reflect uniform induction of cells. As the toluene concentration increases, the level of gfp expression per cell increases until saturation kinetics of the TbuT-PtbuA1 system are observed. Each inducer had a unique minimum concentration that was necessary for induction, with Kapp values that ranged from 3.3 ± 1.8 μM for toluene to 35.6 ± 16.6 μM for trichloroethylene (means ± standard errors of the means), and maximal fluorescence response. The fluorescence response was specific for alkyl-substituted benzene derivatives and branched alkenes (di- and trichloroethylene, 2-methyl-2-butene). The biosensor responded in an additive fashion to the presence of multiple inducers and was unaffected by the presence of compounds that were not inducers, such as those present in gasoline. Flow cytometry revealed that, in response to toxic concentrations of gasoline, there was a small uninduced population and another larger fully induced population whose levels of fluorescence corresponded to the amount of effectors present in the sample. These results demonstrate the potential for green fluorescent protein-based bacterial biosensors to measure environmental contaminants. PMID:11916719

TRACE ANALYSIS OF FLUORESCEIN-DERIVATIZED PHENOXY ACID HERBICIDES BY MICELLAR ELECTROKINETIC CHROMATOGRAPHY WITH LASER-INDUCTED FLUORESCENCE DETECTION

EPA Science Inventory

Micellar electrokinetic chromatography (MEKC) with laser-induced fluorescence (LIF) detection was used for the trace analysis of phenoxy acid herbicides. Capillary electrophoresis (CE) with LIF detection, which has not previously been used for pesticide analysis, overcomes the po...

Heme oxygenase attenuates angiotensin II-mediated superoxide production in cultured mouse thick ascending loop of Henle cells.

PubMed

Kelsen, Silvia; Patel, Bijal J; Parker, Lawson B; Vera, Trinity; Rimoldi, John M; Gadepalli, Rama S V; Drummond, Heather A; Stec, David E

2008-10-01

Heme oxygenase (HO)-1 induction can attenuate the development of angiotensin II (ANG II)-dependent hypertension. However, the mechanism by which HO-1 lowers blood pressure is not clear. The goal of this study was to test the hypothesis that induction of HO-1 can reduce the ANG II-mediated increase in superoxide production in cultured thick ascending loop of Henle (TALH) cells. Studies were performed on an immortalized cell line of mouse TALH (mTALH) cells. HO-1 was induced in cultured mTALH cells by treatment with cobalt protoporphyrin (CoPP, 10 microM) or hemin (50 microM) or by transfection with a plasmid containing the human HO-1 isoform. Treatment of mTALH cells with 10(-9) M ANG II increased dihydroethidium (DHE) fluorescence (an index of superoxide levels) from 35.5+/-5 to 136+/-18 relative fluorescence units (RFU)/microm2. Induction of HO-1 via CoPP, hemin, or overexpression of the human HO-1 isoform significantly reduced ANG II-induced DHE fluorescence to 64+/-5, 64+/-8, and 41+/-4 RFU/microm2, respectively. To determine which metabolite of HO-1 is responsible for reducing ANG II-mediated increases in superoxide production in mTALH cells, cells were preincubated with bilirubin or carbon monoxide (CO)-releasing molecule (CORM)-A1 (each at 100 microM) before exposure to ANG II. DHE fluorescence averaged 80+/-7 RFU/microm2 after incubation with ANG II and was significantly decreased to 55+/-7 and 53+/-4 RFU/microm2 after pretreatment with bilirubin and CORM-A1. These results demonstrate that induction of HO-1 in mTALH cells reduces the levels of ANG II-mediated superoxide production through the production of both bilirubin and CO.

[Effects of NaCl stress on photosynthesis characteristics and fast chlorophyll fluorescence induction dynamics of Pistacia chinensis leaves].

PubMed

Li, Xu-Xin; Liu, Bing-Xiang; Guo, Zhi-Tao; Chang, Yue-Xia; He, Lei; Chen, Fang; Lu, Bing-She

2013-09-01

By using fast chlorophyll fluorescence induction dynamics analysis technique (JIP-test), this paper studied the photosynthesis characteristics and fast chlorophyll fluorescence induction dynamics of 1-year old Pistacia chinensis seedlings under the stress of NaCl at the concentrations 0% (CK), 0.15%, 0.3%, 0.45%, and 0.6%. With the increasing concentration of NaCl, the contents of Chl a, Chl b, and Chl (a+b) in the seedlings leaves decreased, the Chl a/b ratio decreased after an initial increase, and the carotenoid content increased. The net photosynthetic rate (P(n)) and stomatal conductance (g(s)) decreased gradually with increasing NaCl concentration. The decrease of P(n) was mainly attributed to the stomatal limitation when the NaCl concentration was lower than 0.3%, and to the non-stomatal limitation when the NaCl concentration was higher than 0.3%. The trapped energy flux per RC (TR0/CS0), electron transport flux per RC (ET0/CS0), density of RCs (RC/CS0), and yield or flux ratio (psi(0) or phi(E0)) decreased, but the absorption flux per CS (ABS/CS0) and the K phase (W(k)) and J phase (V) in the O-J-I-P chlorophyll fluorescence induction curves increased distinctly, indicating that NaCl stress damaged the leaf oxygen-evolving complex (OEC), donor sides, and PS II reaction centers. When the NaCl concentration reached 0.3%, the maximum photochemical efficiency (F(v)/F(m)) and performance index (PI(ABS)) decreased 17.7% and 36.6%, respectively, as compared with the control.

Determination of major elements by wavelength-dispersive X-ray fluorescence spectrometry and trace elements by inductively coupled plasma mass spectrometry in igneous rocks from the same fused sample (110 mg)

NASA Astrophysics Data System (ADS)

Amosova, Alena A.; Panteeva, Svetlana V.; Chubarov, Victor M.; Finkelshtein, Alexandr L.

2016-08-01

The fusion technique is proposed for simultaneous determination of 35 elements from the same sample. Only 110 mg of rock sample was used to obtain fused glasses for quantitative determination of 10 major elements by wavelength dispersive X-ray fluorescence analysis, 16 rare earth elements and some other trace elements by inductively coupled plasma mass spectrometry analysis. Fusion was performed with 1.1 g of lithium metaborate and LiBr solution as the releasing agent in platinum crucible in electric furnace at 1100 °C. The certified reference materials of ultramafic, mafic, intermediate and felsic igneous rocks have been applied to obtain the calibration curves for rock-forming oxides (Na2O, MgO, Al2O3, SiO2, P2O5, K2O, CaO, TiO2, MnO, Fe2O3) and some trace elements (Ba, Sr, Zr) determination by X-ray fluorescence analysis. The repeatability does not exceed the allowable standard deviation for a wide range of concentrations. In the most cases the relative standard deviation was less than 5%. Obtained glasses were utilized for the further determination of rare earth (La, Ce, Pr, Nd, Sm, Eu, Gd, Tb, Dy, Ho, Er, Tm, Yb, Lu) and some other (Ba, Sr, Zr, Rb, Cs, Y, Nb, Hf, Ta, Th and U) trace elements by inductively coupled plasma mass spectrometry analysis with the same certified reference materials employed. The results could mostly be accepted as satisfactory. The proposed procedure essentially reduces the expenses in comparison with separate sample preparation for inductively coupled plasma mass spectrometry and X-ray fluorescence analysis.

P700+- and 3P700-induced quenching of the fluorescence at 760 nm in trimeric Photosystem I complexes from the cyanobacterium Arthrospira platensis.

PubMed

Schlodder, Eberhard; Cetin, Marianne; Byrdin, Martin; Terekhova, Irina V; Karapetyan, Navassard V

2005-01-07

The 5 K absorption spectrum of Photosystem I (PS I) trimers from Arthrospira platensis (old name: Spirulina platensis) exhibits long-wavelength antenna (exciton) states absorbing at 707 nm (called C707) and at 740 nm (called C740). The lowest energy state (C740) fluoresces around 760 nm (F760) at low temperature. The analysis of the spectral properties (peak position and line width) of the lowest energy transition (C740) as a function of temperature within the linear electron-phonon approximation indicates a large optical reorganization energy of approximately 110 cm(-1) and a broad inhomogeneous site distribution characterized by a line width of approximately 115 cm(-1). Linear dichroism (LD) measurements indicate that the transition dipole moment of the red-most state is virtually parallel to the membrane plane. The relative fluorescence yield at 760 nm of PS I with P700 oxidized increases only slightly when the temperature is lowered to 77 K, whereas in the presence of reduced P700 the fluorescence yield increases nearly 40-fold at 77 K as compared to that at room temperature (RT). A fluorescence induction effect could not be resolved at RT. At 77 K the fluorescence yield of PS I trimers frozen in the dark in the presence of sodium ascorbate decreases during illumination by about a factor of 5 due to the irreversible formation of (P700+)F(A/B-) in about 60% of the centers and the reversible accumulation of the longer-lived state (P700+)FX-. The quenching efficiency of different functionally relevant intermediate states of the photochemistry in PS I has been studied. The redox state of the acceptors beyond A(0) does not affect F760. Direct kinetic evidence is presented that the fluorescence at 760 nm is strongly quenched not only by P700+ but also by 3P700. Similar kinetics were observed for flash-induced absorbance changes attributed to the decay of 3P700 or P700+, respectively, and flash-induced fluorescence changes at 760 nm measured under identical conditions. A nonlinear relationship between the variable fluorescence around 760 nm and the [P700red]/[P700total] ratio was derived from titration curves of the absorbance change at 826 nm and the variable fluorescence at 760 nm as a function of the redox potential imposed on the sample solution at room temperature before freezing. The result indicates that the energy exchange between the antennae of different monomers within a PS I trimer stimulates quenching of F760 by P700+.

Monitoring protein turnover during phosphate starvation-dependent autophagic degradation using a photoconvertible fluorescent protein aggregate in tobacco BY-2 cells

PubMed Central

Tasaki, Maiko; Asatsuma, Satoru; Matsuoka, Ken

2014-01-01

We have developed a system for quantitative monitoring of autophagic degradation in transformed tobacco BY-2 cells using an aggregate-prone protein comprised of cytochrome b5 (Cyt b5) and a tetrameric red fluorescent protein (RFP). Unfortunately, this system is of limited use for monitoring the kinetics of autophagic degradation because the proteins synthesized before and after induction of autophagy cannot be distinguished. To overcome this problem, we developed a system using kikume green-red (KikGR), a photoconvertible and tetrameric fluorescent protein that changes its fluorescence from green to red upon irradiation with purple light. Using the fusion protein of Cyt b5 and KikGR together with a method for the bulk conversion of KikGR, which we had previously used to convert the Golgi-localized monomeric KikGR fusion protein, we were able to monitor both the growth and de novo formation of aggregates. Using this system, we found that tobacco cells do not cease protein synthesis under conditions of phosphate (Pi)-starvation. Induction of autophagy under Pi-starvation, but not under sugar- or nitrogen-starvation, was specifically inhibited by phosphite, which is an analog of Pi with a different oxidation number. Therefore, the mechanism by which BY-2 cells can sense Pi-starvation and induce autophagy does not involve sensing a general decrease in energy supply and a specific Pi sensor might be involved in the induction of autophagy under Pi-starvation. PMID:24817874

Monitoring protein turnover during phosphate starvation-dependent autophagic degradation using a photoconvertible fluorescent protein aggregate in tobacco BY-2 cells.

PubMed

Tasaki, Maiko; Asatsuma, Satoru; Matsuoka, Ken

2014-01-01

We have developed a system for quantitative monitoring of autophagic degradation in transformed tobacco BY-2 cells using an aggregate-prone protein comprised of cytochrome b5 (Cyt b5) and a tetrameric red fluorescent protein (RFP). Unfortunately, this system is of limited use for monitoring the kinetics of autophagic degradation because the proteins synthesized before and after induction of autophagy cannot be distinguished. To overcome this problem, we developed a system using kikume green-red (KikGR), a photoconvertible and tetrameric fluorescent protein that changes its fluorescence from green to red upon irradiation with purple light. Using the fusion protein of Cyt b5 and KikGR together with a method for the bulk conversion of KikGR, which we had previously used to convert the Golgi-localized monomeric KikGR fusion protein, we were able to monitor both the growth and de novo formation of aggregates. Using this system, we found that tobacco cells do not cease protein synthesis under conditions of phosphate (Pi)-starvation. Induction of autophagy under Pi-starvation, but not under sugar- or nitrogen-starvation, was specifically inhibited by phosphite, which is an analog of Pi with a different oxidation number. Therefore, the mechanism by which BY-2 cells can sense Pi-starvation and induce autophagy does not involve sensing a general decrease in energy supply and a specific Pi sensor might be involved in the induction of autophagy under Pi-starvation.

Concentration of Elements in Food: How Can It Reflect Impact of Environmental and Other Influencing Factors?

NASA Astrophysics Data System (ADS)

Vincevica-Gaile, Zane; Klavins, Maris

2013-12-01

Element content of food is variable and can be influenced by different factors. The aim of the present study was to discover the linkage between macro- and microelement concentrations in food produced in Latvia, and possible impacts of environmental factors. More than 300 fresh food samples such as eggs, cottage cheese, honey, root vegetables, apple juice, apple wine were collected in the time period from 2009 to 2011. Samples were mineralised or analysed directly by appropriate method of quantitative analysis: atomic absorption spectrometry, inductively coupled plasma mass spectrometry or total reflection X-ray fluorescence spectrometry. Statistical analysis of data revealed that food elemental content can be influenced by sitespecific factors such as geographical origin, seasonality, environmental pollution.

Stochasticity in the signalling network of a model microbe

NASA Astrophysics Data System (ADS)

Bischofs, Ilka; Foley, Jonathan; Battenberg, Eric; Fontaine-Bodin, Lisa; Price, Gavin; Wolf, Denise; Arkin, Adam

2007-03-01

The soil dwelling bacterium Bacillus subtilis is an excellent model organism for studying stochastic stress response induction in an isoclonal population. Subjected to the same stressor cells undergo different cell fates, including sporulation, competence, degradative enzyme synthesis and motility. For example, under conditions of nutrient deprivation and high cell density only a portion of the cell population forms an endospore. Here we use a combined experimental and theoretical approach to study stochastic sporulation induction in Bacillus subtilis. Using several fluorescent reporter strains we apply time lapse fluorescent microscopy in combination with quantitative image analysis to study cell fate progression on a single cell basis and elucidate key noise generators in the underlying cellular network.

Correlation between lifetime heterogeneity and kinetics heterogeneity during chlorophyll fluorescence induction in leaves: 2. Multi-frequency phase and modulation analysis evidences a loosely connected PSII pigment-protein complex.

PubMed

Moise, Nicolae; Moya, Ismaël

2004-06-28

We report the first direct decomposition of the fluorescence lifetime heterogeneity during multiphasic fluorescence induction in dark-adapted leaves by multi-frequency phase and modulation fluorometry (PMF). A very fast component, assigned to photosystem I (PSI), was found to be constant in lifetime and yield, whereas the two slow components, which are strongly affected by the closure of the reaction centers by light, were assigned to PSII. Based on a modified "reversible radical pair" kinetic model with three compartments, we showed that a loosely connected pigment complex, which is assumed to be the CP47 complex, plays a specific role with respect to the structure and function of the PSII: (i) it explains the heterogeneity of PSII fluorescence lifetime as a compartmentation of excitation energy in the antenna, (ii) it is the site of a conformational change in the first second of illumination, and (iii) it is involved in the mechanisms of nonphotochemical quenching (NPQ). On the basis of the multi-frequency PMF analysis, we reconciled two apparently antagonistic aspects of chlorophyll a fluorescence in vivo: it is heterogeneous with respect to the kinetic structure (several lifetime components) and homogeneous with respect to average quantities (quasi-linear mean tau-Phi relationship).

On the origin of the slow M-T chlorophyll a fluorescence decline in cyanobacteria: interplay of short-term light-responses.

PubMed

Bernát, Gábor; Steinbach, Gábor; Kaňa, Radek; Govindjee; Misra, Amarendra N; Prašil, Ondřej

2018-05-01

The slow kinetic phases of the chlorophyll a fluorescence transient (induction) are valuable tools in studying dynamic regulation of light harvesting, light energy distribution between photosystems, and heat dissipation in photosynthetic organisms. However, the origin of these phases are not yet fully understood. This is especially true in the case of prokaryotic oxygenic photoautotrophs, the cyanobacteria. To understand the origin of the slowest (tens of minutes) kinetic phase, the M-T fluorescence decline, in the context of light acclimation of these globally important microorganisms, we have compared spectrally resolved fluorescence induction data from the wild type Synechocystis sp. PCC 6803 cells, using orange (λ = 593 nm) actinic light, with those of mutants, ΔapcD and ΔOCP, that are unable to perform either state transition or fluorescence quenching by orange carotenoid protein (OCP), respectively. Our results suggest a multiple origin of the M-T decline and reveal a complex interplay of various known regulatory processes in maintaining the redox homeostasis of a cyanobacterial cell. In addition, they lead us to suggest that a new type of regulatory process, operating on the timescale of minutes to hours, is involved in dissipating excess light energy in cyanobacteria.

Drought is Coming: Monitoring Vegetation Response to Water Scarcity through Variable Chlorophyll a Fluorescence

NASA Astrophysics Data System (ADS)

Guadagno, C. R.; Beverly, D.; Pleban, J. R.; Speckman, H. N.; Ewers, B. E.; Weinig, C.

2017-12-01

Aridity is one of the most pronounced environmental limits to plant survival, and understanding how plants respond to drought and recovery is crucial for predicting impacts on managed and natural ecosystems. Changes in soil moisture conditions induce a suite of physiological responses from the cell to ecosystem scale, complicating the assessment of drought effects. Characterizing early indicators of water scarcity across species can inform biophysical models with improved understanding of plant hydraulics. While indexes exist for drought monitoring across scales, many are unable to identify imminent vegetative drought. We explore a method of early diagnosis using leaf-level and kinetic imaging measures of variable chlorophyll a fluorescence. This is a fast and reliable tool capturing leaf physiological changes in advance of changes in NDVI or passive solar induced fluorescence. Both image and leaf level Pulse Amplitude Method (PAM) measurements illustrate the utility of variable chlorophyll a fluorescence for monitoring vegetative drought. Variable fluorescence was monitored across populations of crops, desert shrubs, montane conifers and riparian deciduous trees under variable water regimes. We found a strong correlation (R = 0.85) between the maximum efficiency of photosystem II measured using variable fluorescence (Fv'Fm') and leaf level electrolyte leakage, a proximal cause of drought stress induced by cellular damage in leaves. This association was confirmed in two gymnosperm species (Picea engelmannii and Pinus contorta) and for diverse varieties of the crop species Brassica rapa. The use of chlorophyll a fluorescence per image also allowed for early detection of drought in aspen (Populus tremuloides). These results provide evidence that variable chlorophyll fluorescence decreases between 25% and 70% in mild and severely droughted twigs with respect to ones collected from trees in wet soil conditions. While current systems for monitoring variable fluorescence are limited in scale, chlorophyll fluorescence comprises an indicator of drought stress across multiple spatial scales, from leaf to ecosystem level.

lac operon induction in Escherichia coli: Systematic comparison of IPTG and TMG induction and influence of the transacetylase LacA.

PubMed

Marbach, Anja; Bettenbrock, Katja

2012-01-01

Most commonly used expression systems in bacteria are based on the Escherichia coli lac promoter. Furthermore, lac operon elements are used today in systems and synthetic biology. In the majority of the cases the gratuitous inducers IPTG or TMG are used. Here we report a systematic comparison of lac promoter induction by TMG and IPTG which focuses on the aspects inducer uptake, population heterogeneity and a potential influence of the transacetylase, LacA. We provide induction curves in E. coli LJ110 and in isogenic lacY and lacA mutant strains and we show that both inducers are substrates of the lactose permease at low inducer concentrations but can also enter cells independently of lactose permease if present at higher concentrations. Using a gfp reporter strain we compared TMG and IPTG induction at single cell level and showed that bimodal induction with IPTG occurred at approximately ten-fold lower concentrations than with TMG. Furthermore, we observed that lac operon induction is influenced by the transacetylase, LacA. By comparing two Plac-gfp reporter strains with and without a lacA deletion we could show that in the lacA(+) strain the fluorescence level decreased after few hours while the fluorescence further increased in the lacA(-) strain. The results indicate that through the activity of LacA the IPTG concentration can be reduced below an inducing threshold concentration-an influence that should be considered if low inducer amounts are used. Copyright © 2011 Elsevier B.V. All rights reserved.

The causes of altered chlorophyll fluorescence quenching induction in the Arabidopsis mutant lacking all minor antenna complexes.

PubMed

Townsend, Alexandra J; Saccon, Francesco; Giovagnetti, Vasco; Wilson, Sam; Ungerer, Petra; Ruban, Alexander V

2018-03-13

Non-photochemical quenching (NPQ) of chlorophyll fluorescence is the process by which excess light energy is harmlessly dissipated within the photosynthetic membrane. The fastest component of NPQ, known as energy-dependent quenching (qE), occurs within minutes, but the site and mechanism of qE remain of great debate. Here, the chlorophyll fluorescence of Arabidopsis thaliana wild type (WT) plants was compared to mutants lacking all minor antenna complexes (NoM). Upon illumination, NoM exhibits altered chlorophyll fluorescence quenching induction (i.e. from the dark-adapted state) characterised by three different stages: (i) a fast quenching component, (ii) transient fluorescence recovery and (iii) a second quenching component. The initial fast quenching component originates in light harvesting complex II (LHCII) trimers and is dependent upon PsbS and the formation of a proton gradient across the thylakoid membrane (ΔpH). Transient fluorescence recovery is likely to occur in both WT and NoM plants, but it cannot be overcome in NoM due to impaired ΔpH formation and a reduced zeaxanthin synthesis rate. Moreover, an enhanced fluorescence emission peak at ~679 nm in NoM plants indicates detachment of LHCII trimers from the bulk antenna system, which could also contribute to the transient fluorescence recovery. Finally, the second quenching component is triggered by both ΔpH and PsbS and enhanced by zeaxanthin synthesis. This study indicates that minor antenna complexes are not essential for qE, but reveals their importance in electron stransport, ΔpH formation and zeaxanthin synthesis. Copyright © 2018 The Author(s). Published by Elsevier B.V. All rights reserved.

Improved circuit for measuring capacitive and inductive reactances

NASA Technical Reports Server (NTRS)

Dalins, I.; Mc Carty, V.

1967-01-01

Amplifier circuit measures very small changes of capacitive or inductive reactance, such as produced by a variable capacitance or a variable inductance displacement transducer. The circuit employs reactance-sensing oscillators in which field effect transistors serve as the active elements.

A Sensitive Sensor Cell Line for the Detection of Oxidative Stress Responses in Cultured Human Keratinocytes

PubMed Central

Hofmann, Ute; Priem, Melanie; Bartzsch, Christine; Winckler, Thomas; Feller, Karl-Heinz

2014-01-01

In the progress of allergic and irritant contact dermatitis, chemicals that cause the generation of reactive oxygen species trigger a heat shock response in keratinocytes. In this study, an optical sensor cell line based on cultured human keratinocytes (HaCaT cells) expressing green fluorescent protein (GFP) under the control of the stress-inducible HSP70B' promoter were constructed. Exposure of HaCaT sensor cells to 25 μM cadmium, a model substance for oxidative stress induction, provoked a 1.7-fold increase in total glutathione and a ∼300-fold induction of transcript level of the gene coding for heat shock protein HSP70B'. An extract of Arnica montana flowers resulted in a strong induction of the HSP70B' gene and a pronounced decrease of total glutathione in keratinocytes. The HSP70B' promoter-based sensor cells conveniently detected cadmium-induced stress using GFP fluorescence as read-out with a limit of detection of 6 μM cadmium. In addition the sensor cells responded to exposure of cells to A. montana extract with induction of GFP fluorescence. Thus, the HaCaT sensor cells provide a means for the automated detection of the compromised redox status of keratinocytes as an early indicator of the development of human skin disorders and could be applied for the prediction of skin irritation in more complex in vitro 3D human skin models and in the development of micro-total analysis systems (μTAS) that may be utilized in dermatology, toxicology, pharmacology and drug screenings. PMID:24967604

The promoter of the cereal VERNALIZATION1 gene is sufficient for transcriptional induction by prolonged cold.

PubMed

Alonso-Peral, Maria M; Oliver, Sandra N; Casao, M Cristina; Greenup, Aaron A; Trevaskis, Ben

2011-01-01

The VERNALIZATION1 (VRN1) gene of temperate cereals is transcriptionally activated by prolonged cold during winter (vernalization) to promote flowering. To investigate the mechanisms controlling induction of VRN1 by prolonged cold, different regions of the VRN1 gene were fused to the GREEN FLUORESCENT PROTEIN (GFP) reporter and expression of the resulting gene constructs was assayed in transgenic barley (Hordeum vulgare). A 2 kb segment of the promoter of VRN1 was sufficient for GFP expression in the leaves and shoot apex of transgenic barley plants. Fluorescence increased at the shoot apex prior to inflorescence initiation and was subsequently maintained in the developing inflorescence. The promoter was also sufficient for low-temperature induction of GFP expression. A naturally occurring insertion in the proximal promoter, which is associated with elevated VRN1 expression and early flowering in some spring wheats, did not abolish induction of VRN1 transcription by prolonged cold, however. A translational fusion of the promoter and transcribed regions of VRN1 to GFP, VRN1::GFP, was localised to nuclei of cells at the shoot apex of transgenic barley plants. The distribution of VRN1::GFP at the shoot apex was similar to the expression pattern of the VRN1 promoter-GFP reporter gene. Fluorescence from the VRN1::GFP fusion protein increased in the developing leaves after prolonged cold treatment. These observations suggest that the promoter of VRN1 is targeted by mechanisms that trigger vernalization-induced flowering in economically important temperate cereal crops.

Time-lapse monitoring of TLR2 ligand internalization with newly developed fluorescent probes.

PubMed

Arai, Yohei; Yokoyama, Kouhei; Kawahara, Yuki; Feng, Qi; Ohta, Ippei; Shimoyama, Atsushi; Inuki, Shinsuke; Fukase, Koichi; Kabayama, Kazuya; Fujimoto, Yukari

2018-05-23

As a mammalian toll-like receptor family member protein, TLR2 recognizes lipoproteins from bacteria and modulates the immune response by inducing the expression of various cytokines. We have developed fluorescence-labeled TLR2 ligands with either hydrophilic or hydrophobic fluorescence groups. The labeled ligands maintained the inflammatory IL-6 induction activity and enabled us to observe the internalization and colocalization of the TLR2 ligands using live-cell imaging. The time-lapse monitoring in the live-cell imaging of the fluorescence-labeled TLR2 ligand showed that TLR2/CD14 expression in the host cells enhanced the internalization of TLR2 ligand molecules.

XRF inductive bead fusion and PLC based control system

NASA Astrophysics Data System (ADS)

Zhu, Jin-hong; Wang, Ying-jie; Shi, Hong-xin; Chen, Qing-ling; Chen, Yu-xi

2009-03-01

In order to ensure high-quality X-ray fluorescence spectrometry (XRF) analysis, an inductive bead fusion machine was developed. The prototype consists of super-audio IGBT induction heating power supply, rotation and swing mechanisms, and programmable logic controller (PLC). The system can realize sequence control, mechanical movement control, output current and temperature control. Experimental results show that the power supply can operate at an ideal quasi-resonant state, in which the expected power output and the required temperature can be achieved for rapid heating and the uniform formation of glass beads respectively.

Altered impulse activity modifies synaptic physiology and mitochondria in crayfish phasic motor neurons.

PubMed

Nguyen, P V; Atwood, H L

1994-12-01

1. Crayfish phasic motor synapses produce large initial excitatory postsynaptic potentials (EPSPs) that fatigue rapidly during high-frequency stimulation. Periodic in vivo stimulation of an identified phasic abdominal extensor motor neuron (axon 3) induced long-term adaptation (LTA) of neuromuscular transmission: initial EPSP amplitude became smaller and synaptic depression was significantly reduced. We tested the hypothesis that activity-induced synaptic fatigue-resistance seen during LTA was dependent upon, or correlated with, mitochondrial oxidative competence. 2. Periodic unilateral conditioning stimulation of axon 3 entering each of two adjacent homologous abdominal segments (segments 2 and 3) increased the synaptic stamina in both "conditioned" axons; mean final EPSP amplitudes, recorded after 20 min of 5-Hz test stimulation, were significantly larger than those measured with the same protocol from contralateral unstimulated axons. 3. During 5-Hz test stimulation of the conditioned axon 3 of segment 3, acute superfusion with 0.8 mM dinitrophenol or 20 mM sodium azide [inhibitors of oxidative adenosinetriphosphate (ATP) synthesis] produced increased synaptic depression. Drug-free saline superfusion of the conditioned axon 3 of segment 2 in these same animals did not affect the increased synaptic fatigue resistance seen in this segment. Thus both successful induction (in axon 3 of saline-perfused segment 2) and attenuation (in axon 3 of drug-perfused segment 3) of the increased synaptic stamina can be demonstrated with this twin-segment conditioning protocol. 4. Confocal microscopic imaging of mitochondrial rhodamine-123 (Rh123) fluorescence was used to assess relative oxidative competence of conditioned and unconditioned phasic axons. Conditioned phasic axons showed significantly higher mean mitochondrial Rh123 fluorescence than contralateral unstimulated axons. In the same preparations that showed increased postconditioning Rh123 fluorescence, the synaptic fatigue resistance measured from conditioned axon 3 was also significantly greater than that recorded from contralateral unstimulated axon 3. 5. Axotomy of the phasic extensor nerve root (containing axon 3), before in vivo conditioning stimulation of its decentralized segment, prevented induction of both the increased synaptic stamina in axon 3 and the enhanced mitochondrial fluorescence in decentralized motor axons of the nerve root. Hence, induction of both changes requires axonal transport of materials between the soma and the motor synapses of axon 3. 5. Axotomy of the phasic extensor nerve root (containing axon 3), before in vivo conditioning stimulation of its decentralized segment, Prevented induction of both the increased synaptic stamina in axon 3 and the enhanced mitochondrial fluorescence in decentralized motor axons of the nerve root Hence, induction of both changes requires axonal transport of materials between the soma and the motor synapses of axon 3 6. Because mitochondrial Rh123 fluorescence is primarily dependent upon the oxidative activity of these organelles, our findings suggest that conditioning stimulation of phasic extensor axon 3 increases its mitochondrial oxidative competence and that the enhanced synaptic stamina seen during LTA in axon 3 is correlated with, and dependent upon, oxidative activity.(ABSTRACT TRUNCATED AT 400 WORDS)

Enhancing Induction Coil Reliability

NASA Astrophysics Data System (ADS)

Kreter, K.; Goldstein, R.; Yakey, C.; Nemkov, V.

2014-12-01

In induction hardening, thermal fatigue is one of the main copper failure modes of induction heat treating coils. There have been papers published that describe this failure mode and others that describe some good design practices. The variables previously identified as the sources of thermal fatigue include radiation from the part surface, frequency, current, concentrator losses, water pressure and coil wall thickness. However, there is very little quantitative data on the factors that influence thermal fatigue in induction coils is available in the public domain. By using finite element analysis software this study analyzes the effect of common design variables of inductor cooling, and quantifies the relative importance of these variables. A comprehensive case study for a single shot induction coil with Fluxtrol A concentrator applied is used for the analysis.

Live Dynamics of 53BP1 Foci Following Simultaneous Induction of Clustered and Dispersed DNA Damage in U2OS Cells

PubMed Central

Sollazzo, Alice; Brzozowska, Beata; Cheng, Lei; Lundholm, Lovisa; Scherthan, Harry

2018-01-01

Cells react differently to clustered and dispersed DNA double strand breaks (DSB). Little is known about the initial reaction to simultaneous induction of DSBs with different complexities. Here, we used live cell microscopy to analyse the behaviour of 53BP1-GFP (green fluorescence protein) foci formation at DSBs induced in U2OS cells by alpha particles, X-rays or mixed beams over a 75 min period post irradiation. X-ray-induced foci rapidly increased and declined over the observation interval. After an initial increase, mixed beam-induced foci remained at a constant level over the observation interval, similarly as alpha-induced foci. The average areas of radiation-induced foci were similar for mixed beams and X-rays, being significantly smaller than those induced by alpha particles. Pixel intensities were highest for mixed beam-induced foci and showed the lowest level of variability over time as compared to foci induced by alphas and X-rays alone. Finally, mixed beam-exposed foci showed the lowest level of mobility as compared to alpha and X-ray exposure. The results suggest paralysation of chromatin around foci containing clustered DNA damage. PMID:29419809

Preliminary study of effects of military obscurant smokes on flora and fauna during field and laboratory exposures. Final technical report

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

Schaeffer, D.J.; Lower, W.R.; Kapila, S.

1986-12-01

Since continued routine use of obscurant smokes could be detrimental to the native flora and fauna of training sites, a preliminary biological and chemical study of smokes was conducted to determine whether tests could be developed to demonstrate measurable changes in organisms exposed to smokes and to evaluate whether short exposures to smokes produced measurable changes in the organisms tested. Fog oil, hexachloroethane, and tank diesel smokes were tested. Tradescantia clones were examined for mutagenic effects indicated by micronuclei induction in developing pollen and pink somatic mutations in stamen hairs. Photosynthetic perturbations were measured in Tradescantia and Ambrosia dumosa usingmore » variable fluorescence induction. Animals were examined for sister chromatid exchanges and chromosome aberrations. It was found that all of the smokes tested exerted varying degrees of physiological and mutagenic effects in one or several of the assay systems at one or more of the exposure distances. These studies indicate that exposed ecological systems, or at least components of these systems, are at a higher risk than are control organisms for several types of damage attributed to obscurant smoke exposure.« less

A T-DNA insertion mutant of AtHMA1 gene encoding a Cu transporting ATPase in Arabidopsis thaliana has a defect in the water-water cycle of photosynthesis.

PubMed

Higuchi, Mieko; Ozaki, Hiroshi; Matsui, Minami; Sonoike, Kintake

2009-03-03

The water-water cycle is the electron flow through scavenging enzymes for the reactive species of oxygen in chloroplasts, and is proposed to play a role in alternative electron sink in photosynthesis. Here we showed that the water-water cycle is impaired in the T-DNA insertion mutant of AtHMA1 gene encoding a Cu transporting ATPase in chloroplasts. Chlorophyll fluorescence under steady state was not affected in hma1, indicating that photosynthetic electron transport under normal condition was not impaired. Under electron acceptor limited conditions, however, hma1 showed distinguished phenotype in chlorophyll fluorescence characteristics. The most severe phenotype of hma1 could be observed in high (0.1%) CO(2) concentrations, indicating that hma1 has the defect other than photorespiration. The transient increase of chlorophyll fluorescence upon the cessation of the actinic light as well as the NPQ induction of chlorophyll fluorescence revealed that the two pathways of cyclic electron flow around PSI, NDH-pathway and FQR-pathway, are both intact in hma1. Based on the NPQ induction under 0% oxygen condition, we conclude that the water-water cycle is impaired in hma1, presumably due to the decreased level of Cu/Zn SOD in the mutant. Under high CO(2) condition, hma1 exhibited slightly higher NPQ induction than wild type plants, while this increase of NPQ in hma1 was suppressed when hma1 was crossed with crr2 having a defect in NDH-mediated PSI cyclic electron flow. We propose that the water-water cycle and NDH-mediated pathways might be regulated compensationally with each other especially when photorespiration is suppressed.

PHYTOPLANKTON NUTRIENT STATUS AND VARIABLE FLUORESCENCE MEASUREMENTS IN A GULF COAST ESTUARY

EPA Science Inventory

Changes in variable fluorescence parameters such as the maximum quantum yield of fluorescence (a.k.a. photosynthetic efficiency and Fv/Fm) have been related to nutrient status in single-species cultures. To test if changes in Fv/Fm of mixed natural assemblages were related to nut...

Induction of apoptosis in human promyelocytic leukemia HL-60 cells by Ampelopsis cantoniensis crude extract.

PubMed

Tan, Tzu-Wei; Tsai, Huei-Yann; Chen, Yuh-Fung; Chung, Jing-Gung

2004-01-01

The crude extract of Ampelopsis cantoniensis induced apoptosis in human promyelocytic leukemia HL-60 cells and this induction was investigated by flow cytometric analysis, DNA gel electrophoresis and poly (ADP-ribose) fluorescence staining. The results demonstrated that this extract induced dose-dependent cytotoxicity and apoptosis. The level of active caspase-3 was increased after treatment with the crude extract for 24 hours.

Portable real-time fluorescence cytometry of microscale cell culture analog devices

NASA Astrophysics Data System (ADS)

Kim, Donghyun; Tatosian, Daniel A.; Shuler, Michael L.

2006-02-01

A portable fluorescence cytometric system that provides a modular platform for quantitative real-time image measurements has been used to explore the applicability to investigating cellular events on multiple time scales. For a short time scale, we investigated the real-time dynamics of uptake of daunorubicin, a chemotherapeutic agent, in cultured mouse L-cells in a micro cell culture analog compartment using the fluorescent cytometric system. The green fluorescent protein (GFP) expression to monitor induction of pre-specified genes, which occurs on a much longer time scale, has also been measured. Here GFP fluorescence from a doxycycline inducible promoter in a mouse L-cell line was determined. Additionally, a system based on inexpensive LEDs showed performance comparable to a broadband light source based system and reduced photobleaching compared to microscopic examination.

Highly selective and sensitive detection of Cu2+ with lysine enhancing bovine serum albumin modified-carbon dots fluorescent probe.

PubMed

Liu, Jia-Ming; Lin, Li-ping; Wang, Xin-Xing; Lin, Shao-Qin; Cai, Wen-Lian; Zhang, Li-Hong; Zheng, Zhi-Yong

2012-06-07

Based on the ability of lysine (Lys) to enhance the fluorescence intensity of bovine serum albumin modified-carbon dots (CDs-BSA) to decrease surface defects and quench fluorescence of the CDs-BSA-Lys system in the presence of Cu(2+) under conditions of phosphate buffer (PBS, pH = 5.0) at 45 °C for 10 min, a sensitive Lys enhancing CDs-BSA fluorescent probe was designed. The environment-friendly, simple, rapid, selective and sensitive fluorescent probe has been utilized to detect Cu(2+) in hair and tap water samples and it achieved consistent results with those obtained by inductively coupled plasma mass spectroscopy (ICP-MS). The mechanism of the proposed assay for the detection of Cu(2+) is discussed.

The onset of anthracene phototoxicity to Lemna gibba and the protective effects of humic acid

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

Gensemer, R.W.; Dixon, D.G.; Greenberg, B.M.

1994-12-31

The toxicity of anthracene to the freshwater duckweed Lemna gibba is strongly photo-induced in the presence of light containing natural levels of ultraviolet (UV) radiation. This was demonstrated using 8-day static renewal bioassays at an anthracene concentration of 2 mg-L{sup {minus}1}. Plants were incubated under simulated solar radiation (SSR) which mimics UV levels found in natural sunlight at a visible:UV-A:UV-B ratio of 100:10:1. Anthracene phototoxicity was expressed as inhibition of population growth and fluorescence induction decreases in chlorophyll content, and changes in low-temperature chlorophyll fluorescence emission scans. Furthermore, adding 6.2 mg-L-1 of an artificial humic acid ameliorated anthracene phototoxicity evenmore » though HA is also photo modified by UV light. However, anthracene inhibited photosynthesis days before the endpoint assays were performed. Therefore, the authors repeated these experiments at short time intervals following exposure to both light and chemical. Anthracene phototoxicity occurred after only 1 hour as detected by chlorophyll fluorescence induction, whereas chlorophyll contents and low-temperature fluorescence emission scans were not affected until 24--48 hours, respectively. Humic acid again ameliorated anthracene toxicity by delaying the negative physiological events by approximately 24 hours.« less

Comparative study of atomic fluorescence spectroscopy and inductively coupled plasma mass spectrometry for mercury and arsenic multispeciation.

PubMed

Gómez-Ariza, José Luis; Lorenzo, Fernando; García-Barrera, Tamara

2005-05-01

Mercury and arsenic are two elements of undoubted importance owing to their toxic character. Although speciation of these elements has been developed separately, in this work for the first time the speciation of As and Hg using two atomic fluorescence detectors in a sequential ensemble is presented. A coupling based on the combination of high-performance liquid chromatography (where mercury and arsenic species are separated) and two atomic fluorescence detectors in series, with several online treatments, including photooxidation (UV) and hydride generation, has allowed the determination of mercury and arsenic compounds simultaneously. The detection limits for this device were 16, 3, 17, 12 and 8 ng mL(-1) for As(III), monomethylarsinic acid, As(V), Hg2+ and methylmercury, respectively. This coupling was compared with an analogous one based on inductively coupled plasma-mass spectrometry (ICP-MS) detection, with detection limits of 0.7, 0.5, 0.8, 0.9 and 1.1 ng mL(-1), respectively. Multispeciation based on ICP-MS exhibits better sensitivity than the coupling based on tandem atomic fluorescence, but this second device is a very robust system and exhibits obvious advantages related to the low cost of acquisition and maintenance, as well as easy handling, which makes it a suitable system for routine laboratories.

High temporal resolution fluorescence measurements of a mitochondrial dye for detection of early stage apoptosis

PubMed Central

Iyer, Divya; Ray, Rachel D.; Pappas, Dimitri

2013-01-01

In the present study, early stage apoptosis is explored with high temporal resolution. In addition to monitoring early apoptosis induction in single cells by ultrasensitive confocal fluorescence microscopy (UCFM), the mitochondrial proteins release kinetics was explored. The current study shows development and optimization of a novel, rapid apoptosis assay to explore the earliest changes in cells by the intrinsic apoptosis pathway. We show that early apoptotic changes in the mitochondria begin nearly simultaneously with the addition of an apoptosis-inducing drug, such as staurosporine. With a temporal resolution of five minutes, this non-invasive analytical technique can elucidate the earliest apoptotic events in living cells. Moreover, our results show that the mitochondrial inter-membrane proteins are not involved in the extrinsic pathway of Ramos cells mediated by an anti-CD95 antibody. Additional techniques such as light microscopy and flow cytometry were employed to confirm the results obtained by ultrasensitive confocal fluorescence microscopy. The results of this study help to understand the earliest mechanisms of apoptosis induction in cells, enabling new methods of drug testing and dose-response analyses. PMID:23831722

Variable fluorescence parameters in the filamentous Patagonian rhodophytes, Callithamnion gaudichaudii and Ceramium sp. under solar radiation.

PubMed

Häder, Donat-P; Lebert, Michael; Helbling, E Walter

2004-01-23

The filamentous rhodophytes Callithamnion gaudichaudi Agardh and Ceramium sp. were utilized to study the effects of solar radiation (PAR, 400-700 nm, UV-B, 280-315 nm and UV-A, 315-400 nm) on the photosynthetic performance in situ in Patagonia waters (Argentina). A pulse amplitude modulated (PAM) fluorometer was used to determine the fluorescence parameters. The two species grew in different habitats in the eulittoral: Ceramium sp. was found only in rock pools while C. gaudichaudii grew on exposed rocks and fell dry during low tide. Both species differed in their fluorescence parameters and their sensitivity to solar radiation exposure. The photosynthetic quantum yield had its lowest values at noon, but it recovered in the afternoon/evening hours, when irradiances were lower. PAR (irradiance of about 400 W m(-2) at noon) was responsible for most of the decrease in the yield on clear days, especially in Ceramium sp., but UVR (280-400 nm) also accounted for a significant decrease. Fluence rate response curves indicated that both species were adapted to low fluence rates and showed a pronounced non-photochemical quenching at intermediate and higher irradiances. Both species showed a rapid adaptation during measurement of fast induction kinetics but differed significantly in their fluorescence components. All photosynthetic pigments were bleached after 8 h exposure to solar radiation over a full day. Strong absorption in the UV-A range, most likely due to mycosporine-like amino acids, was detected in both strains. The pronounced sensitivity to solar radiation in situ and the recovery capacity of these two filamentous Rhodophyte species, as well as the presence of protective compounds, suggests that these algae have the ability to adapt to the relatively high radiation levels and changes in irradiance found in the Patagonia waters.

Freeze-Fracture Ultrastructure of Thylakoid Membranes in Chloroplasts from Manganese-Deficient Plants

PubMed Central

Simpson, David J.; Robinson, Simon P.

1984-01-01

Leaves from spinach (Spinacia oleracea L. cv Hybrid 102) plants grown in Mn-deficient nutrient solution were characterized by chlorosis, lowered chlorophyll a/b ratio and reduced electron transport. There were characteristic changes in room temperature fluorescence induction kinetics with increased initial yield (Fo) and decreased variable fluorescence (Fv). The fluorescence yield after the maximum fell rapidly to a level below Fo. The shape of the rise from Fo to the maximum was altered and the size of photosystem II units increased, as measured by half-rise time of Fv in the presence of 3-(3,4-dichlorophenyl)-1,1-dimethylurea. The Mn-deficient leaves were harvested before necrosis, when thin section electron microscopy revealed no disorganization of the thylakoid system. Thylakoid membranes were examined by freeze-fracture electron microscopy. The effect of Mn-deficiency was the specific loss of three-quarters of the particles from the endoplasmic fracture face of appressed thylakoids (EFs). Mn-deficient leaves were restored to near normal 2 days after application of exogenous Mn to the nutrient solution. It is concluded that the loss of most, but not all, functional photosystem II reaction centers from grana, with no alteration in light-harvesting complex or photosystem I, is responsible for the fluorescence and functional properties observed. The response of thylakoids to Mn deficiency shows that there is a fundamental difference in composition and function of stacked and unstacked endoplasmic fracture particles. The stacked endoplasmic fracture particle probably contains, in close association, the photosystem II reaction center and also the Mn-containing polypeptide, the 3-(3,4-dichlorophenyl)-1,1-dimethylurea-binding protein, and all electron transport components in between. Images Fig. 3 Fig. 4 Fig. 5 PMID:16663491

Temporal variability in chlorophyll fluorescence of back-reef corals in Ofu, American Samoa

USGS Publications Warehouse

Piniak, G.A.; Brown, E.K.

2009-01-01

Change in the yield of chlorophyll a fluorescence is a common indicator of thermal stress in corals. The present study reports temporal variability in quantum yield measurements for 10 coral species in Ofu, American Samoa - a place known to experience elevated and variable seawater temperatures. In winter, the zooxanthellae generally had higher dark-adapted maximum quantum yield (F v/Fm), higher light- adapted effective quantum yield (??F/F'm), and lower relative electron transport rates (rETR) than in the summer. Temporal changes appeared unrelated to the expected bleaching sensitivity of corals. All species surveyed, with the exception of Montipora grisea, demonstrated significant temporal changes in the three fluorescence parameters. Fluorescence responses were influenced by the microhabitat - temporal differences in fluorescence parameters were usually observed in the habitat with a more variable temperature regime (pool 300), while differences in Fv/Fm between species were observed only in the more environmentally stable habitat (pool 400). Such species-specific responses and microhabitat variability should be considered when attempting to determine whether observed in situ changes are normal seasonal changes or early signs of bleaching. ?? 2009 Marine Biological Laboratory.

Pharmacokinetics and pharmacodynamics of propofol with or without 2% benzyl alcohol following a single induction dose administered intravenously in cats.

PubMed

Griffenhagen, Gregg M; Rezende, Marlis L; Gustafson, Daniel L; Hansen, Ryan J; Lunghofer, Paul J; Mama, Khursheed R

2015-09-01

To compare the pharmacokinetics and pharmacodynamics of propofol with or without 2% benzyl alcohol administered intravenously (IV) as a single induction dose in cats. Prospective experimental study. Six healthy adult cats, three female intact, three male castrated, weighing 4.8 ± 1.8 kg. Cats received 8 mg kg(-1) IV of propofol (P) or propofol with 2% benzyl alcohol (P28) using a randomized crossover design. Venous blood samples were collected at predetermined time points to 24 hours after drug administration to determine drug plasma concentrations. Physiologic and behavioral variables were also recorded. Propofol and benzyl alcohol concentrations were determined using high pressure liquid chromatography with fluorescence detection. Pharmacokinetic parameters were described using a 2-compartment model. Pharmacokinetic and pharmacodynamic parameters were analyzed using repeated measures anova (p < 0.05). Plasma concentrations of benzyl alcohol were below the lower limits of quantification (LLOQ) at all time points for two of the six cats (33%), and by 30 minutes for the remaining four cats. Propofol pharmacokinetics, with or without 2% benzyl alcohol, were characterized by rapid distribution, a long elimination phase, and a large volume of distribution. No differences were noted between treatments with the exception of clearance from the second compartment (CLD2), which was 23.6 and 38.8 mL kg(-1)  minute(-1) in the P and P28 treatments, respectively. Physiologic and behavioral variables were not different between treatments with the exception of heart rate at 4 hours post administration. The addition of 2% benzyl alcohol as a preservative minimally altered the pharmacokinetics and pharmacodynamics of propofol 1% emulsion when administered as a single IV bolus in this group of cats. These data support the cautious use of propofol with 2% benzyl alcohol for induction of anesthesia in healthy cats. © 2014 Association of Veterinary Anaesthetists and the American College of Veterinary Anesthesia and Analgesia.

Participation of intracellular and extracellular pH changes in photosynthetic response development induced by variation potential in pumpkin seedlings.

PubMed

Sherstneva, O N; Vodeneev, V A; Katicheva, L A; Surova, L M; Sukhov, V S

2015-06-01

Electrical signals presented in plants by action potential and by variation potential (VP) can induce a reversible inactivation of photosynthesis. Changes in the intracellular and extracellular pH during VP generation are a potential mechanism of photosynthetic response induction; however, this hypothesis requires additional experimental investigation. The purpose of the present work was to analyze the influence of pH changes on induction of the photosynthetic response in pumpkin. It was shown that a burning of the cotyledon induced VP propagation into true leaves of pumpkin seedlings inducing a decrease in the photosynthetic CO2 assimilation and an increase in non-photochemical quenching of fluorescence, whereas respiration was activated insignificantly. The photosynthetic response magnitude depended linearly on the VP amplitude. The intracellular and extracellular concentrations of protons were analyzed using pH-sensitive fluorescent probes, and the VP generation was shown to be accompanied by apoplast alkalization (0.4 pH unit) and cytoplasm acidification (0.3 pH unit). The influence of changes in the incubation medium pH on the non-photochemical quenching of fluorescence of isolated chloroplasts was also investigated. It was found that acidification of the medium stimulated the non-photochemical quenching, and the magnitude of this increase depended on the decrease in pH. Our results confirm the contribution of changes in intracellular and extracellular pH to induction of the photosynthetic response caused by VP. Possible mechanisms of the influence of pH changes on photosynthesis are discussed.

Autophagy induction by silibinin positively contributes to its anti-metastatic capacity via AMPK/mTOR pathway in renal cell carcinoma.

PubMed

Li, Feng; Ma, Zhenkun; Guan, Zhenfeng; Chen, Yule; Wu, Kaijie; Guo, Peng; Wang, Xinyang; He, Dalin; Zeng, Jin

2015-04-15

Silibinin, a dietary cancer chemopreventive flavonoid from the seeds of milk thistle, has been reported to exhibit anti-metastatic effects on renal cell carcinoma (RCC), but the mechanism underlying this phenomenon is not fully understood. The present study aimed at examining the potential role of autophagy in regulating silibinin-induced anti-metastatic effects on RCC cells. Using RCC ACHN and 786-O cells as a model system in vitro, we found that silibinin treatment increased the expression of LC3-II, resulted in the formation of autophagolysosome vacuoles, and caused a punctate fluorescence pattern with the monomeric red fluorescence protein-enhanced green fluorescence protein-LC3 (mRFP-EGFP-LC3) protein, which all are markers for cellular autophagy. Autophagy flux was induced by silibinin in RCC cells, as determined by LC3 turnover assay. Mechanically, the adenosine 5'-monophosphate activated protein kinase (AMPK)/mammalian target of rapamycin (mTOR) pathway was identified as involved in regulation of silibinin-induced autophagy. Furthermore, autophagy induction was demonstrated to positively contribute to silibinin-induced anti-metastatic effects on RCC cells in vitro. Activation of autophagy enhanced silibinin-induced inhibition of migration and invasion of RCC cells, while inhibition of autophagy attenuated it. These findings thus provide new information about the potential link between autophagy and metastasis inhibition induced by silibinin, and the induction of autophagy may shed some light into future treatment strategies for metastatic RCC.

Autophagy Induction by Silibinin Positively Contributes to Its Anti-Metastatic Capacity via AMPK/mTOR Pathway in Renal Cell Carcinoma

PubMed Central

Li, Feng; Ma, Zhenkun; Guan, Zhenfeng; Chen, Yule; Wu, Kaijie; Guo, Peng; Wang, Xinyang; He, Dalin; Zeng, Jin

2015-01-01

Silibinin, a dietary cancer chemopreventive flavonoid from the seeds of milk thistle, has been reported to exhibit anti-metastatic effects on renal cell carcinoma (RCC), but the mechanism underlying this phenomenon is not fully understood. The present study aimed at examining the potential role of autophagy in regulating silibinin-induced anti-metastatic effects on RCC cells. Using RCC ACHN and 786-O cells as a model system in vitro, we found that silibinin treatment increased the expression of LC3-II, resulted in the formation of autophagolysosome vacuoles, and caused a punctate fluorescence pattern with the monomeric red fluorescence protein-enhanced green fluorescence protein-LC3 (mRFP-EGFP-LC3) protein, which all are markers for cellular autophagy. Autophagy flux was induced by silibinin in RCC cells, as determined by LC3 turnover assay. Mechanically, the adenosine 5'-monophosphate activated protein kinase (AMPK)/mammalian target of rapamycin (mTOR) pathway was identified as involved in regulation of silibinin-induced autophagy. Furthermore, autophagy induction was demonstrated to positively contribute to silibinin-induced anti-metastatic effects on RCC cells in vitro. Activation of autophagy enhanced silibinin-induced inhibition of migration and invasion of RCC cells, while inhibition of autophagy attenuated it. These findings thus provide new information about the potential link between autophagy and metastasis inhibition induced by silibinin, and the induction of autophagy may shed some light into future treatment strategies for metastatic RCC. PMID:25884331

In situ investigation of heavy metals at trace concentrations in greenhouse soils via portable X-ray fluorescence spectroscopy.

PubMed

Tian, Kang; Huang, Biao; Xing, Zhe; Hu, Wenyou

2018-04-01

Soil pollution by heavy metals (HMs) has rapidly become a major threat to vegetable security. Nearly all cultivated soils are at risk of metal accumulation, and greenhouse soils are among the most heavily impacted soils. In this study, a rapid assessment of HMs at trace concentrations was conducted via portable X-ray fluorescence (PXRF) spectroscopy in Shouguang, China. Measurements were made via PXRF under in situ, ex situ and sieved conditions and by inductively coupled plasma mass spectrometry (ICP-MS) after acid digestion. The performance of each PXRF measure relative to the ICP-MS method was assessed by linear regression. Redundancy analysis was performed to quantify the proportion of explained variability between the PXRF and ICP-MS data. Evaluation of the possible sources of HMs and their potential risks was then conducted by multivariate analysis. The results showed that the PXRF data were closely correlated with ICP-MS quantification for Cu, Mn and Zn, whereas no significant correlations were found for As, Ni and Pb. The uncertainties of PXRF measurement derived from soil heterogeneity accounted for 20.02% of total variability and those from moisture and particle size accounted for 20.15%. The geo-accumulation index (I geo ) indicated that the greenhouse soils were potentially contaminated by Cu and Zn (I geo  > 0), which can be attributed to anthropogenic activities. Overall, PXRF spectroscopy is promising as a rapid and nondestructive in situ technique for assessing the potential risks of HMs at trace concentrations in greenhouse soils.

The Covert World of Fish Biofluorescence: A Phylogenetically Widespread and Phenotypically Variable Phenomenon

PubMed Central

Schelly, Robert C.; Smith, W. Leo; Davis, Matthew P.; Tchernov, Dan; Pieribone, Vincent A.

2014-01-01

The discovery of fluorescent proteins has revolutionized experimental biology. Whereas the majority of fluorescent proteins have been identified from cnidarians, recently several fluorescent proteins have been isolated across the animal tree of life. Here we show that biofluorescence is not only phylogenetically widespread, but is also phenotypically variable across both cartilaginous and bony fishes, highlighting its evolutionary history and the possibility for discovery of numerous novel fluorescent proteins. Fish biofluorescence is especially common and morphologically variable in cryptically patterned coral-reef lineages. We identified 16 orders, 50 families, 105 genera, and more than 180 species of biofluorescent fishes. We have also reconstructed our current understanding of the phylogenetic distribution of biofluorescence for ray-finned fishes. The presence of yellow long-pass intraocular filters in many biofluorescent fish lineages and the substantive color vision capabilities of coral-reef fishes suggest that they are capable of detecting fluoresced light. We present species-specific emission patterns among closely related species, indicating that biofluorescence potentially functions in intraspecific communication and evidence that fluorescence can be used for camouflage. This research provides insight into the distribution, evolution, and phenotypic variability of biofluorescence in marine lineages and examines the role this variation may play. PMID:24421880

The covert world of fish biofluorescence: a phylogenetically widespread and phenotypically variable phenomenon.

PubMed

Sparks, John S; Schelly, Robert C; Smith, W Leo; Davis, Matthew P; Tchernov, Dan; Pieribone, Vincent A; Gruber, David F

2014-01-01

The discovery of fluorescent proteins has revolutionized experimental biology. Whereas the majority of fluorescent proteins have been identified from cnidarians, recently several fluorescent proteins have been isolated across the animal tree of life. Here we show that biofluorescence is not only phylogenetically widespread, but is also phenotypically variable across both cartilaginous and bony fishes, highlighting its evolutionary history and the possibility for discovery of numerous novel fluorescent proteins. Fish biofluorescence is especially common and morphologically variable in cryptically patterned coral-reef lineages. We identified 16 orders, 50 families, 105 genera, and more than 180 species of biofluorescent fishes. We have also reconstructed our current understanding of the phylogenetic distribution of biofluorescence for ray-finned fishes. The presence of yellow long-pass intraocular filters in many biofluorescent fish lineages and the substantive color vision capabilities of coral-reef fishes suggest that they are capable of detecting fluoresced light. We present species-specific emission patterns among closely related species, indicating that biofluorescence potentially functions in intraspecific communication and evidence that fluorescence can be used for camouflage. This research provides insight into the distribution, evolution, and phenotypic variability of biofluorescence in marine lineages and examines the role this variation may play.

Spatial variability of oceanic phycoerythrin spectral types derived from airborne laser-induced fluorescence emissions

NASA Astrophysics Data System (ADS)

Hoge, Frank E.; Wright, C. Wayne; Kana, Todd M.; Swift, Robert N.; Yungel, James K.

1998-07-01

We report spatial variability of oceanic phycoerythrin spectral types detected by means of a blue spectral shift in airborne laser-induced fluorescence emission. The blue shift of the phycoerythrobilin fluorescence is known from laboratory studies to be induced by phycourobilin chromophore substitution at phycoerythrobilin chromophore sites in some strains of phycoerythrin-containing marine cyanobacteria. The airborne 532-nm laser-induced phycoerythrin fluorescence of the upper oceanic volume showed distinct segregation of cyanobacterial chromophore types in a flight transect from coastal water to the Sargasso Sea in the western North Atlantic. High phycourobilin levels were restricted to the oceanic (oligotrophic) end of the flight transect, in agreement with historical ship findings. These remotely observed phycoerythrin spectral fluorescence shifts have the potential to permit rapid, wide-area studies of the spatial variability of spectrally distinct cyanobacteria, especially across interfacial regions of coastal and oceanic water masses. Airborne laser-induced phytoplankton spectral fluorescence observations also further the development of satellite algorithms for passive detection of phytoplankton pigments. Optical modifications to the NASA Airborne Oceanographic Lidar are briefly described that permitted observation of the fluorescence spectral shifts.

Linking fluorescence induction curve and biomass in herbicide screening.

PubMed

Christensen, Martin G; Teicher, Harald B; Streibig, Jens C

2003-12-01

A suite of dose-response bioassays with white mustard (Sinapis alba L) and sugar beet (Beta vulgaris L) in the greenhouse and with three herbicides was used to analyse how the fluorescence induction curves (Kautsky curves) were affected by the herbicides. Bentazone, a photosystem II (PSII) inhibitor, completely blocked the normal fluorescence decay after the P-step. In contrast, fluorescence decay was still obvious for flurochloridone, a PDS inhibitor, and glyphosate, an EPSP inhibitor, which indicated that PSII inhibition was incomplete. From the numerous parameters that can be derived from OJIP-steps of the Kautsky curve the relative changes at the J-step [Fvj = (Fm - Fj)/Fm] was selected to be a common response parameter for the herbicides and yielded consistent dose-response relationships. Four hours after treatment, the response Fvj on the doses of bentazone and flurochloridone could be measured. For glyphosate, the changes of the Kautsky curve could similarly be detected 4 h after treatment in sugar beet, but only after 24 hs in S alba. The best prediction of biomass in relation to Fvj was found for bentazone. The experiments were conducted between May and August 2002 and showed that the ambient temperature and solar radiation in the greenhouse could affect dose-response relationships. If the Kautsky curve parameters should be used to predict the outcome of herbicide screening experiments in the greenhouse, where ambient radiation and temperature can only partly be controlled, it is imperative that the chosen fluorescence parameters can be used to predict accurately the resulting biomass used in classical bioassays.

Fluorescence quenching and the "ring-mode" to "red-mode" transition in alkali inductively coupled plasmas

NASA Astrophysics Data System (ADS)

Huang, M.; Bazurto, R.; Camparo, J.

2018-01-01

The ring-mode to red-mode transition in alkali metal inductively coupled plasmas (ICPs) (i.e., rf-discharge lamps) is perhaps the most important physical phenomenon affecting these devices as optical pumping light sources for atomic clocks and magnetometers. It sets the limit on useful ICP operating temperature, thereby setting a limit on ICP light output for atomic-clock/magnetometer signal generation, and it is a temperature region of ICP operation associated with discharge instability. Previous work has suggested that the mechanism driving the ring-mode to red-mode transition is associated with radiation trapping, but definitive experimental evidence validating that hypothesis has been lacking. Based on that hypothesis, one would predict that the introduction of an alkali-fluorescence quenching gas (i.e., N2) into the ICP would increase the ring-mode to red-mode transition temperature. Here, we test that prediction, finding direct evidence supporting the radiation-trapping hypothesis.

Focal Adhesion Induction at the Tip of a Functionalized Nanoelectrode

PubMed Central

Fuentes, Daniela E.; Bae, Chilman; Butler, Peter J.

2012-01-01

Cells dynamically interact with their physical micro-environment through the assembly of nascent focal contacts and focal adhesions. The dynamics and mechanics of these contact points are controlled by transmembrane integrins and an array of intracellular adaptor proteins. In order to study the mechanics and dynamics of focal adhesion assembly, we have developed a technique for the timed induction of a nascent focal adhesion. Bovine aortic endothelial cells were approached at the apical surface by a nanoelectrode whose position was controlled with a resolution of 10s of nanometers using changes in electrode current to monitor distance from the cell surface. Since this probe was functionalized with fibronectin, a focal contact formed at the contact location. Nascent focal adhesion assembly was confirmed using time-lapse confocal fluorescent images of red fluorescent protein (RFP) – tagged talin, an adapter protein that binds to activated integrins. Binding to the cell was verified by noting a lack of change of electrode current upon retraction of the electrode. This study demonstrates that functionalized nanoelectrodes can enable precisely-timed induction and 3-D mechanical manipulation of focal adhesions and the assay of the detailed molecular kinetics of their assembly. PMID:22247742

Detecting crop population growth using chlorophyll fluorescence imaging.

PubMed

Wang, Heng; Qian, Xiangjie; Zhang, Lan; Xu, Sailong; Li, Haifeng; Xia, Xiaojian; Dai, Liankui; Xu, Liang; Yu, Jingquan; Liu, Xu

2017-12-10

For both field and greenhouse crops, it is challenging to evaluate their growth information on a large area over a long time. In this work, we developed a chlorophyll fluorescence imaging-based system for crop population growth information detection. Modular design was used to make the system provide high-intensity uniform illumination. This system can perform modulated chlorophyll fluorescence induction kinetics measurement and chlorophyll fluorescence parameter imaging over a large area of up to 45  cm×34  cm. The system can provide different lighting intensity by modulating the duty cycle of its control signal. Results of continuous monitoring of cucumbers in nitrogen deficiency show the system can reduce the judge error of crop physiological status and improve monitoring efficiency. Meanwhile, the system is promising in high throughput application scenarios.

Polypyrrole-based nanotheranostics for activatable fluorescence imaging and chemo/photothermal dual therapy of triple-negative breast cancer

NASA Astrophysics Data System (ADS)

Park, Dongjin; Ahn, Kyung-Ohk; Jeong, Kyung-Chae; Choi, Yongdoo

2016-05-01

Here, we fabricated polypyrrole nanoparticles (PPys) (termed HA10-PPy, HA20-PPy, and HA40-PPy) doped with different average molecular weight hyaluronic acids (HAs) (10, 20, and 40 kDa, respectively), and evaluated the effect of molecular weight of doped HA on photothermal induction, fluorescence quenching, and drug loading efficiencies. Doxorubicin-loaded HA-doped PPys (DOX@HA-PPys) could be used for imaging and therapy of triple-negative breast cancer (TNBC). Fluorescence turn-on, stimuli-responsive drug release, and photo-induced heating of DOX@HA-PPys enabled not only activatable fluorescence imaging but also subsequent chemo/photothermal dual therapy for TNBC. In particular, we illustrated the potential usefulness of the photothermal effect of the nanoparticles for overcoming chemoresistance in TNBC.

Chemometric analysis for extraction of individual fluorescence spectrum and lifetimes from a target mixture

NASA Technical Reports Server (NTRS)

Hallidy, William H. (Inventor); Chin, Robert C. (Inventor)

1999-01-01

The present invention is a system for chemometric analysis for the extraction of the individual component fluorescence spectra and fluorescence lifetimes from a target mixture. The present invention combines a processor with an apparatus for generating an excitation signal to transmit at a target mixture and an apparatus for detecting the emitted signal from the target mixture. The present invention extracts the individual fluorescence spectrum and fluorescence lifetime measurements from the frequency and wavelength data acquired from the emitted signal. The present invention uses an iterative solution that first requires the initialization of several decision variables and the initial approximation determinations of intermediate matrices. The iterative solution compares the decision variables for convergence to see if further approximation determinations are necessary. If the solution converges, the present invention then determines the reduced best fit error for the analysis of the individual fluorescence lifetime and the fluorescence spectrum before extracting the individual fluorescence lifetime and fluorescence spectrum from the emitted signal of the target mixture.

Fluorescence F 0 of photosystems II and I in developing C3 and C 4 leaves, and implications on regulation of excitation balance.

PubMed

Peterson, Richard B; Oja, Vello; Eichelmann, Hillar; Bichele, Irina; Dall'Osto, Luca; Laisk, Agu

2014-10-01

This work addresses the question of occurrence and function of photosystem II (PSII) in bundle sheath (BS) cells of leaves possessing NADP-malic enzyme-type C4 photosynthesis (Zea mays). Although no requirement for PSII activity in the BS has been established, several component proteins of PSII have been detected in BS cells of developing maize leaves exhibiting O2-insensitive photosynthesis. We used the basal fluorescence emissions of PSI (F 0I) and PSII (F 0II) as quantitative indicators of the respective relative photosystem densities. Chl fluorescence induction was measured simultaneously at 680 and 750 nm. In mature leaves, the F m(680)/F 0(680) ratio was 10.5 but less in immature leaves. We propose that the lower ratio was caused by the presence of a distinct non-variable component, F c, emitting at 680 and 750 nm. After F c was subtracted, the fluorescence of PSI (F 0I) was detected as a non-variable component at 750 nm and was undetectably low at 680 nm. Contents of Chls a and b were measured in addition to Chl fluorescence. The Chl b/(a + b) was relatively stable in developing sunflower leaves (0.25-0.26), but in maize it increased from 0.09 to 0.21 with leaf tissue age. In sunflower, the F 0I/(F 0I + F 0II) was 0.39 ± 0.01 independent of leaf age, but in maize, this parameter was 0.65 in young tissue of very low Chl content (20-50 mg m(-2)) falling to a stable level of 0.53 ± 0.01 at Chl contents >100 mg m(-2). The values of F 0I/(F 0I + F 0II) showed that in sunflower, excitation was partitioned between PSII and PSI in a ratio of 2:1, but the same ratio was 1:1 in the C4 plant. The latter is consistent with a PSII:PSI ratio of 2:1 in maize mesophyll cells and PSI only in BS cells (2:1:1 distribution). We suggest, moreover, that redox mediation of Chl synthesis, rather than protein accumulation, regulates photosystem assembly to ensure optimum excitation balance between functional PSII and PSI. Indeed, the apparent necessity for two Chls (a and b) may reside in their targeted functions in influencing accumulation of PSI and PSII, respectively, as opposed to their spectral differences.

On the inherent competition between valid and spurious inductive inferences in Boolean data

NASA Astrophysics Data System (ADS)

Andrecut, M.

Inductive inference is the process of extracting general rules from specific observations. This problem also arises in the analysis of biological networks, such as genetic regulatory networks, where the interactions are complex and the observations are incomplete. A typical task in these problems is to extract general interaction rules as combinations of Boolean covariates, that explain a measured response variable. The inductive inference process can be considered as an incompletely specified Boolean function synthesis problem. This incompleteness of the problem will also generate spurious inferences, which are a serious threat to valid inductive inference rules. Using random Boolean data as a null model, here we attempt to measure the competition between valid and spurious inductive inference rules from a given data set. We formulate two greedy search algorithms, which synthesize a given Boolean response variable in a sparse disjunct normal form, and respectively a sparse generalized algebraic normal form of the variables from the observation data, and we evaluate numerically their performance.

Energy saving concepts relating to induction generators

NASA Technical Reports Server (NTRS)

Nola, F. J.

1980-01-01

Energy saving concepts relating to induction generators are presented. The first describes a regenerative scheme using an induction generator as a variable load for prime movers under test is described. A method for reducing losses in induction machines used specifically as wind driven generators is also described.

A facile, sensitive, and highly specific trinitrophenol assay based on target-induced synergetic effects of acid induction and electron transfer towards DNA-templated copper nanoclusters.

PubMed

Li, Haiyin; Chang, Jiafu; Hou, Ting; Ge, Lei; Li, Feng

2016-11-01

Reliable, selective and sensitive approaches for trinitrophenol (TNP) detection are highly desirable with respect to national security and environmental protection. Herein, a simple and novel fluorescent strategy for highly sensitive and specific TNP assay has been successfully developed, which is based on the quenching of the fluorescent poly(thymine)-templated copper nanoclusters (DNA-CuNCs), through the synergetic effects of acid induction and electron transfer. Upon the addition of TNP, donor-acceptor complexes between the electron-deficient nitro-groups in TNP and the electron-donating DNA templates are formed, resulting in the close proximity between TNP and CuNCs. Moreover, the acidity of TNP contributes to the pH decrease of the system. These factors combine to dramatically quench the fluorescence of DNA-CuNCs, providing a "signal-off" strategy for TNP sensing. The as-proposed strategy demonstrates high sensitivity for TNP assay, and a detection limit of 0.03μM is obtained, which is lower than those reported by using organic fluorescent materials. More significantly, this approach shows outstanding selectivity over a number of TNP analogues, such as 2,4,6-trinitrotoluene (TNT), 2,4-dinitrotoluene (DNT), 2,4-dinitrophenol (DNP), 3-nitrophenol (NP), nitrobenzene (NB), phenol (BP), and toluene (BT). Compared with previous studies, this method does not need complex DNA sequence design, fluorescent dye labeling, or sophisticated organic reactions, rendering the strategy with additional advantages of simplicity and cost-effectiveness. In addition, the as-proposed strategy has been adopted for the detection of TNP in natural water samples, indicating its great potential to be applied in the fields of public safety and environmental monitoring. Copyright © 2016 Elsevier B.V. All rights reserved.

Xenopus tropicalis transgenic lines and their use in the study of embryonic induction.

PubMed

Hirsch, Nicolas; Zimmerman, Lyle B; Gray, Jessica; Chae, Jeiwook; Curran, Kristen L; Fisher, Marilyn; Ogino, Hajime; Grainger, Robert M

2002-12-01

For over a century, amphibian embryos have been a source of significant insight into developmental mechanisms, including fundamental discoveries about the process of induction. The recently developed transgenesis for Xenopus offers new approaches to these poorly understood processes, particularly when undertaken in the quickly maturing species Xenopus tropicalis, which greatly facilitates establishment of permanent transgenic lines. Several X. tropicalis transgenic lines have now been generated, and experiments demonstrating the value of these lines to study induction in embryonic tissue recombinants and explants are presented here. A revised protocol for transgenesis in X. tropicalis resulting in a significant increase in the percentage of transgenic animals that reach adulthood is presented, as well as improvements in tadpole and froglet husbandry, which have facilitated the raising of large numbers of adults. Working transgenic populations have been rapidly expanded, and some transgenes have been bred to homozygosity. Established lines include those bearing the promoter regions of Pax-6, Otx-2, Rx, and EF1alpha coupled to fluorescent reporter genes. Multireporter lines combining, in a single animal, up to three gene promoters coupled to different fluorescent reporters have also been established. The value of X. tropicalis transgenic lines for the study of induction is demonstrated by showing activation of Pax-6 by noggin treatment of Pax-6/GFP transgenic animal caps, illustrating how reporter lines allow a rapid, in vivo assay for an inductive response. An experiment showing lens induction in gamma-crystallin/GFP transgenic lens ectoderm when it is recombined with mouse optic vesicle demonstrates conservation of inducing signals from amphibians and mammals. It also shows how the warmer culture temperatures tolerated by X. tropicalis embryos can be used in assays of factors produced by mammalian cells and tissues. The many applications of transgenic reporter lines and other lines designed to target gene expression in particular tissues promise to bring significant new insights to the classic issues first defined in amphibian systems. Copyright 2002 Wiley-Liss, Inc.

Induction of Fluorescence from Tremorgenic Mycotoxins

USDA-ARS?s Scientific Manuscript database

There are a large number of neurotoxic mycotoxins, most of which contain an indole-moiety within their structures. A subgroup of such toxins are the tremorgens, which are further divided into several smaller, structurally related groups. Tremorgens have been associated with toxic occurrences in ca...

Variability in chlorophyll fluorescence spectra of eggplant fruit grown under different light environments: a case study.

PubMed

Ospina Calvo, Brian; Parapugna, Tamara L; Lagorio, M Gabriela

2017-05-17

The main goal of the present work was to clarify physiological strategies in plants whose chloroplasts were developed under different light environments. The specific objective was to elucidate the influence of the spectral distribution of light on the chlorophyll fluorescence ratio and on photosynthetic parameters. To achieve this purpose, three species of eggplant fruit (black, purple and white striped and white) were used as a case study and their chlorophyll fluorescence was analyzed in detail. Spectra of the non-variable fluorescence in each part of the fruit were corrected for distortions by light reabsorption processes using a physical model. The main conclusion of this work was that the corrected fluorescence ratio was dependent on the contribution of each photosystem to the fluorescence and consequently on the environmental lighting conditions, becoming higher when illumination was rich in long wavelengths. Variable chlorophyll fluorescence, similar to that observed from plant leaves, was detected for the pulp of the black eggplant, for the pulp of the purple and white striped eggplant and for the intact fruit of the black eggplant. The maximum quantum efficiency of photosystem II in the light-adapted state (F' v /F' m ), the quantum efficiency of photosystem II (Φ PSII ), and the photochemical and non-photochemical quenching coefficients (qP and qNP/NPQ respectively) were determined in each case. The results could be explained very interestingly, in relation with the proportion of exciting light reaching each photosystem (I and II). The photochemical parameters obtained from variable chlorophyll fluorescence, allowed us to monitor non-destructively the physiological state of the black fruit during storage under both chilled or room-temperature conditions.

Relationship between the Fluorescence Lifetime of Chlorophyll 'a' and Primary Productivity within the Mississippi River Plume and Adjacent Shelf Region

NASA Technical Reports Server (NTRS)

Hall, Callie; Miller, Richard L.; Fernandez, Salvador M.; McKee, Brent A.

2000-01-01

In situ measurements of chlorophyll fluorescence intensity have been widely used to estimate phytoplankton biomass. However, because the fluorescence quantum yield of chlorophyll a in vivo can be highly variable, measurements of chlorophyll fluorescence intensity cannot be directly correlated with phytoplankton biomass and do not provide information on the physiological state of the phytoplankton under study. Conversely, lifetime-based measurements of chlorophyll fluorescence provide a framework in which photosynthetic rates of phytoplankton can be analyzed according to phytoplankton physiology. Along with the measurement of primary production and ambient nutrient concentrations within the Mississippi River plume in the northern Gulf of Mexico, phytoplankton fluorescence lifetimes were measured using a Fluorescence Lifetime Phytoplankton Analyzer (developed under a NASA Small Business Innovative Research contract to Ciencia, Inc.). Variability of fluorescence lifetimes within the plume can be used as a background from which to interpret variations in the maximum quantum yield of photochemistry. The extent to which nutrient and effluent loading in this dynamic coastal area affect the photosynthetic performance of phytoplankton will be presented as a function of phytoplankton fluorescence lifetimes.

Étude comparative des techniques d'analyse par fluorescence X à dispersion d'énergie (ED-XRF) et à dispersion de longueur d'onde (WD-XRF), et par spectrométrie d'émission atomique à source plasma couplée par induction (ICP-AES)

NASA Astrophysics Data System (ADS)

Rahmani, A.; Benyaïch, F.; Bounakhla, M.; Bilal, E.; Moutte, J.; Gruffat, J. J.; Zahry, F.

2004-11-01

Dans ce travail, nous présentons une étude comparative des techniques d'analyse par fluorescence X à dispersion d'énergie (ED-XRF) et à dispersion de longueur d'onde (WD-XRF), et par spectrométrie d'émission atomique à source plasma couplée par induction (ICP-AES). Les résultats de la calibration des spectromètres à dispersion d'énergie, à excitation par sources radioactives (55Fe, 109Cd et 241Am) et à excitation secondaire (cible secondaire Mo et Cu) du Centre National pour l'Energie, les Sciences et les Techniques Nucléaires (CNESTEN, Rabat, Maroc) sur des échantillons étalons de références de l'Agence International de l'Energie Atomique (AIEA) et du Community Bureau of Référence (BCR) ont été comparés aux résultats d'analyse des mêmes échantillons étalons par la spectrométrie X à dispersion de longueur d'onde (WD-XRF) et par spectrométrie d'émission atomique à source plasma couplé par induction (ICP-AES) au département GENERIC du centre SPIN à l'Ecole des Mines de Saint-Etienne (France). Les trois techniques d'analyse utilisées donnent des résultats comparables pour le dosage des éléments majeurs, alors que pour les traces on note des déviations importantes à cause des effets de matrice qui sont difficiles à corriger dans le cas de la fluorescence X.

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.

Induction of Parkinson disease-related proteins in motor neurons after transient spinal cord ischemia in rabbits.

PubMed

Sakurai, Masahiro; Kawamura, Takae; Nishimura, Hidekazu; Suzuki, Hiroyoshi; Tezuka, Fumiaki; Abe, Koji

2009-04-01

The mechanism of spinal cord injury has been thought to be related to the vulnerability of spinal motor neuron cells against ischemia. However, the mechanisms of such vulnerability are not fully understood. We investigated a possible mechanism of neuronal death by immunohistochemical analysis for DJ-1, PINK1, and alpha-Synuclein. We used a 15-min rabbit spinal cord ischemia model, with use of a balloon catheter. Western blot analysis for DJ-1, PINK1, and alpha-Synuclein; temporal profiles of DJ-1, PINK1, and alpha-Synuclein immunoreactivity; and double-label fluorescence immunocytochemical studies were performed. Western blot analysis revealed scarce immunoreactivity for DJ-1, PINK1, and alpha-Synuclein in the sham-operated spinal cords. However, they became apparent at 8 h after transient ischemia, which returned to the baseline level at 1 day. Double-label fluorescence immunocytochemical study revealed that both DJ-1 and PINK1, and DJ-1 and alpha-Synuclein were positive at 8 h of reperfusion in the same motor neurons, which eventually die. The induction of DJ-1 and PINK1 proteins in the motor neurons at the early stage of reperfusion may indicate oxidative stress, and the induction of alpha-Synuclein may be implicated in the programmed cell death change after transient spinal cord ischemia.

Novel mouse model of colitis characterized by hapten-protein visualization.

PubMed

Ishiguro, Kazuhiro; Ando, Takafumi; Maeda, Osamu; Watanabe, Osamu; Goto, Hidemi

2010-09-01

Trinitrobenzene sulfonic acid (TNBS) and oxazolone are used to induce colitis for the investigation of inflammatory reactions in the colon. Although these chemicals are presumed to bind proteins in the colonic mucosa and then induce colitis as haptens, hapten-protein formation has not yet been confirmed in the colonic mucosa. We developed a mouse model of colitis characterized by hapten-protein visualization, using 4-chloro-7-nitro-2,1,3-benzoxadiazole (NBD-Cl), which emits fluorescence after binding to proteins. The enema of 1 mg/mL NBD-Cl induced severe diarrhea, rectal bleeding, and body weight reductions in BALB/c mice. Mucosal signs indicative of colitis, such as redness and swelling observed under stereomicroscopy or inflammatory cell infiltration and crypt-epithelium destruction under microscopy, were manifested around NBD-proteins visualized with fluorescence. Fluorescence microscopy showed the infiltration of F4/80+ cells around areas of NBD-proteins, and flow cytometry indicated the uptake of NBD-proteins by CD11b+ cells. We also found critical roles for T cells and interleukin-6 in colitis induction with NBD-proteins. NBD-Cl-induced colitis presents a unique model to study the relevance between hapten-protein formation and inflammatory reactions and offers a method to assess experimental interventions on colitis induction in the mucosa, where hapten-protein formation is confirmed.

Structural reorganization of the fungal endoplasmic reticulum upon induction of mycotoxin biosynthesis

DOE PAGES

Boenisch, Marike Johanne; Broz, Karen Lisa; Purvine, Samuel Owen; ...

2017-03-13

Eukaryotic cells routinely compartmentalize metabolic pathways to particular organelles for biosynthetic purposes. Relatively few studies have addressed the cellular localization of pathways for secondary metabolites synthesis. In this study, the phytopathogenic fungus Fusarium graminearum reorganized its endoplasmic reticulum (ER) when triggered to produce mycotoxins, both in vitro and in planta. Fluorescence tagged biosynthetic proteins were found to co-localize with the modified ER as confirmed by co-fluorescence and co-purification with known ER proteins. Microscopy, cell sorting, and proteomics were applied in this FICUS collaborative effort.

Four quadrant control of induction motors

NASA Technical Reports Server (NTRS)

Hansen, Irving G.

1991-01-01

Induction motors are the nation's workhorse, being the motor of choice in most applications due to their simple rugged construction. It has been estimated that 14 to 27 percent of the country's total electricity use could be saved with adjustable speed drives. Until now, induction motors have not been suited well for variable speed or servo-drives, due to the inherent complexity, size, and inefficiency of their variable speed controls. Work at NASA Lewis Research Center on field oriented control of induction motors using pulse population modulation method holds the promise for the desired drive electronics. The system allows for a variable voltage to frequency ratio which enables the user to operate the motor at maximum efficiency, while having independent control of both the speed and torque of an induction motor in all four quadrants of the speed torque map. Multiple horsepower machine drives were demonstrated, and work is on-going to develop a 20 hp average, 40 hp peak class of machine. The pulse population technique, results to date, and projections for implementation of this existing new motor control technology are discussed.

Induction of Shiga Toxin-Encoding Prophage by Abiotic Environmental Stress in Food.

PubMed

Fang, Yuan; Mercer, Ryan G; McMullen, Lynn M; Gänzle, Michael G

2017-10-01

The prophage-encoded Shiga toxin is a major virulence factor in Stx-producing Escherichia coli (STEC). Toxin production and phage production are linked and occur after induction of the RecA-dependent SOS response. However, food-related stress and Stx-prophage induction have not been studied at the single-cell level. This study investigated the effects of abiotic environmental stress on stx expression by single-cell quantification of gene expression in STEC O104:H4 Δ stx2 :: gfp :: amp r In addition, the effect of stress on production of phage particles was determined. The lethality of stressors, including heat, HCl, lactic acid, hydrogen peroxide, and high hydrostatic pressure, was selected to reduce cell counts by 1 to 2 log CFU/ml. The integrity of the bacterial membrane after exposure to stress was measured by propidium iodide (PI). The fluorescent signals of green fluorescent protein (GFP) and PI were quantified by flow cytometry. The mechanism of prophage induction by stress was evaluated by relative gene expression of recA and cell morphology. Acid (pH < 3.5) and H 2 O 2 (2.5 mM) induced the expression of stx 2 in about 18% and 3% of the population, respectively. The mechanism of prophage induction by acid differs from that of induction by H 2 O 2 H 2 O 2 induction but not acid induction corresponded to production of infectious phage particles, upregulation of recA , and cell filamentation. Pressure (200 MPa) or heat did not induce the Stx2-encoding prophage (Stx2-prophage). Overall, the quantification method developed in this study allowed investigation of prophage induction and physiological properties at the single-cell level. H 2 O 2 and acids mediate different pathways to induce Stx2-prophage. IMPORTANCE Induction of the Stx-prophage in STEC results in production of phage particles and Stx and thus relates to virulence as well as the transduction of virulence genes. This study developed a method for a detection of the induction of Stx-prophages at the single-cell level; membrane permeability and an indication of SOS response to environmental stress were additionally assessed. H 2 O 2 and mitomycin C induced expression of the prophage and activated a SOS response. In contrast, HCl and lactic acid induced the Stx-prophage but not the SOS response. The lifestyle of STEC exposes the organism to intestinal and extraintestinal environments that impose oxidative and acid stress. A more thorough understanding of the influence of food processing-related stressors on Stx-prophage expression thus facilitates control of STEC in food systems by minimizing prophage induction during food production and storage. Copyright © 2017 American Society for Microbiology.

Induction of Shiga Toxin-Encoding Prophage by Abiotic Environmental Stress in Food

PubMed Central

Fang, Yuan; Mercer, Ryan G.; McMullen, Lynn M.

2017-01-01

ABSTRACT The prophage-encoded Shiga toxin is a major virulence factor in Stx-producing Escherichia coli (STEC). Toxin production and phage production are linked and occur after induction of the RecA-dependent SOS response. However, food-related stress and Stx-prophage induction have not been studied at the single-cell level. This study investigated the effects of abiotic environmental stress on stx expression by single-cell quantification of gene expression in STEC O104:H4 Δstx2::gfp::ampr. In addition, the effect of stress on production of phage particles was determined. The lethality of stressors, including heat, HCl, lactic acid, hydrogen peroxide, and high hydrostatic pressure, was selected to reduce cell counts by 1 to 2 log CFU/ml. The integrity of the bacterial membrane after exposure to stress was measured by propidium iodide (PI). The fluorescent signals of green fluorescent protein (GFP) and PI were quantified by flow cytometry. The mechanism of prophage induction by stress was evaluated by relative gene expression of recA and cell morphology. Acid (pH < 3.5) and H2O2 (2.5 mM) induced the expression of stx2 in about 18% and 3% of the population, respectively. The mechanism of prophage induction by acid differs from that of induction by H2O2. H2O2 induction but not acid induction corresponded to production of infectious phage particles, upregulation of recA, and cell filamentation. Pressure (200 MPa) or heat did not induce the Stx2-encoding prophage (Stx2-prophage). Overall, the quantification method developed in this study allowed investigation of prophage induction and physiological properties at the single-cell level. H2O2 and acids mediate different pathways to induce Stx2-prophage. IMPORTANCE Induction of the Stx-prophage in STEC results in production of phage particles and Stx and thus relates to virulence as well as the transduction of virulence genes. This study developed a method for a detection of the induction of Stx-prophages at the single-cell level; membrane permeability and an indication of SOS response to environmental stress were additionally assessed. H2O2 and mitomycin C induced expression of the prophage and activated a SOS response. In contrast, HCl and lactic acid induced the Stx-prophage but not the SOS response. The lifestyle of STEC exposes the organism to intestinal and extraintestinal environments that impose oxidative and acid stress. A more thorough understanding of the influence of food processing-related stressors on Stx-prophage expression thus facilitates control of STEC in food systems by minimizing prophage induction during food production and storage. PMID:28778890

Dynamometer Research Facilities | Wind | NREL

Science.gov Websites

drivetrains by replacing the rotor and blades of a turbine with a powerful motor. The National Renewable -horsepower variable-speed induction motor, with AC grid connections of 120, 240, and 480 volts (V) and a dynamometer features a 3,351-horsepower (hp), 415-amp AC induction motor with variable-frequency drive that

Changes in Beliefs about Category Homogeneity and Variability across Childhood

ERIC Educational Resources Information Center

Brandone, Amanda C.

2017-01-01

Effective category-based induction requires understanding that categories include both fundamental similarities between members and important variation. This article explores 4- to 11-year-olds' (n = 207) and adults' (n = 49) intuitions about this balance between within-category homogeneity and variability using a novel induction task in which…

Confocal Microscopy for the Histological Fluorescence Pattern of a Recurrent Atypical Meningioma: Case Report

PubMed Central

Whitson, Wesley J.; Valdes, Pablo A.; Harris, Brent T.; Paulsen, Keith D.; Roberts, David W.

2013-01-01

Background and Importance Fluorescence-guided resection with 5-aminolevulinic acid (5-ALA), which has shown promising results in the resection of malignant gliomas, has been used for meningioma resection in an attempt to more clearly delineate the tumor margin. However, no article has investigated the fluorescence pattern of meningiomas on a histological level. Understanding the microscopic pattern of fluorescence could help assess the precision and utility of using 5-ALA for these tumors. We present the case of a recurrent atypical meningioma operated on with 5-ALA fluorescence-guided resection for delineation of tumor tissue from surrounding uninvolved dura. Clinical Presentation A 53-year-old woman presented with recurrent atypical meningioma of the falx. Prior treatment included surgical resection 6 years earlier with subsequent fractionated radiation therapy and radiosurgery for tumor progression. The patient was given 5-ALA 20 mg/kg body weight dissolved in 100 mL water 3 hours before induction of anesthesia. Intraoperative fluorescence was coregistered with preoperative imaging. Neuropathological analysis of the resected falx with confocal microscopy enabled correlation of fluorescence with the extent of tumor on a histological level. Conclusion Fluorescence guidance allowed clear intraoperative delineation of tumor tissue from adjacent, uninvolved dura. On a microscopic level, there was a very close correlation of fluorescence with tumor, but some tumor cells did not fluoresce. PMID:21389893

Noncontact point spectroscopy guided by two-channel fluorescence imaging in a hamster cheek pouch model

NASA Astrophysics Data System (ADS)

Yang, Victor X.; Yeow, Jenny; Lilge, Lothar D.; Kost, James; Mang, Thomas S.; Wilson, Brian C.

1999-07-01

A system for in vivo, fluorescence image-guided, non-contact point fluorescence spectroscopy is presented. A 442 nm HeCd laser is used as the fluorescence excitation source. An intensified CCD serves as the detector for both imaging and spectroscopy, on which two regions of 300 X 300 pixels were used for green (500 +/- 18 nm) and red (630 +/- 18 nm) imaging channels, and a strip of 600 X 120 pixels are used for emission spectroscopy (450 - 750 nm). At a working distance of 40 mm, the system has a spatial resolution of 0.16 mm and a spectral resolution of 5 nm. System performance is demonstrated in a carcinogenesis model in hamsters, where tumors were induced by painting DMBA in the cheek pouch. Autofluorescence and Photofrin-induced fluorescence measurements were performed every 2 weeks during the 18 weeks of tumor induction. Punch biopsies on selected animals were taken for histological staging. The results show that autofluorescence fluorescence can distinguish dysplasia from normal mucosal tissue model, utilizing the peak red intensity (or the red-to-green intensity ratio). Photofrin-induced fluorescence was superior to autofluorescence for differentiating high grade dysplasia from invasive cancer.

Fluorescence detection of small gastrointestinal tumours: principles, technique, first clinical experience.

PubMed

Orth, K; Russ, D; Steiner, R; Beger, H G

2000-11-01

Photodynamic therapy (PDT) is a form of cancer treatment based on the selective accumulation of a photosensitizer in neoplastic tissue. The fluorescent properties of a photosensitizer permit diagnostic localization of primary tumours and/or metastasis. Occult lesions are hard to detect and can easily be missed during routine laparoscopy. Fluorescence observation offers additional optical information and the ability to detect these occult tumours. Clinically, we used 5-aminolevulinic acid for peritoneal staining and tumour demarcation via tumour-specific fluorescence induced by protoporphyrin IX. For laparoscopic observations, a "D-Light" system was used; the conventional white light source was equipped with an optical blocking filter that transmits at the excitation wavelength (380-450 nm) and blocks all other parts of the spectrum. With the aid of a suitable observation filter, the relevant fluorescence was detectable. With the help of this fluorescence we increased the capacity to detect occult tumours, that were missed with white-light observation (9/26). In the gastrointestinal tract, we used a krypton laser at 405 nm for PP IX fluorescence induction. Although there were high sensitivity rates for neoplasms (81% peritoneal carcinomas, 60% gastric cancer), no exact histopathological statement could be achieved at because of false-positive fluorescence, mainly caused by inflammation (6/32). Current clinical goals and the future perspectives of photodynamic diagnostic are discussed.

Effects of chemical smokes on flora and fauna under field and laboratory exposures

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

Schaeffer, D.J.; Novak, E.W.; Lower, W.R.

1987-06-01

Various types of obscurant smokes are used routinely in training by the U.S. Army. Because continued routine use of the smokes could be detrimental to the native flora and fauna at training sites, a preliminary biological and chemical field study of fogoil, hexachloroethane, and tank diesel smokes was conducted. Smoke plumes were sampled and chemically analyzed at distances of 15-150 m from the smoke source where Tradescantia clones 4430 and 03 and the native plant Ambrosia dumosa and the native rodent Dipodomys merriami were exposed for 30 min. In addition, Tradescantia clone 4430 was exposed to tank diesel in themore » laboratory at concentration levels equivalent to exposure at 15 and 50 m. Tradescantia clones were examined for mutagenic effects indicated by micronuclei induction in developing pollen and pink somatic mutations in stamen hairs. Photosynthetic perturbations were measured in Tradescantia and A. dumosa using variable fluorescence induction. Animals were examined for sister chromatid exchanges and chromosome aberrations. It was found that all of the smokes tested exerted varying degrees of physiological and mutagenic effects in one or more assay system at one or more exposure distance. The studies reported here indicate that exposed ecological systems, or at least components of these systems, are at a higher risk than are unexposed components (e.g., organisms) for several types of damage attributed to obscurant smoke exposure.« less

Effects of chemical smokes on flora and fauna under field and laboratory exposures.

PubMed

Schaeffer, D J; Novak, E W; Lower, W R; Yanders, A; Kapila, S; Wang, R

1987-06-01

Various types of obscurant smokes are used routinely in training by the U.S. Army. Because continued routine use of the smokes could be detrimental to the native flora and fauna at training sites, a preliminary biological and chemical field study of fogoil, hexachloroethane, and tank diesel smokes was conducted. Smoke plumes were sampled and chemically analyzed at distances of 15-150 m from the smoke source where Tradescantia clones 4430 and 03 and the native plant Ambrosia dumosa and the native rodent Dipodomys merriami were exposed for 30 min. In addition, Tradescantia clone 4430 was exposed to tank diesel in the laboratory at concentration levels equivalent to exposure at 15 and 50 m. Tradescantia clones were examined for mutagenic effects indicated by micronuclei induction in developing pollen and pink somatic mutations in stamen hairs. Photosynthetic perturbations were measured in Tradescantia and A. dumosa using variable fluorescence induction. Animals were examined for sister chromatid exchanges and chromosome aberrations. It was found that all of the smokes tested exerted varying degrees of physiological and mutagenic effects in one or more assay system at one or more exposure distance. The studies reported here indicate that exposed ecological systems, or at least components of these systems, are at a higher risk than are unexposed components (e.g., organisms) for several types of damage attributed to obscurant smoke exposure.

Robo-Lector - a novel platform for automated high-throughput cultivations in microtiter plates with high information content.

PubMed

Huber, Robert; Ritter, Daniel; Hering, Till; Hillmer, Anne-Kathrin; Kensy, Frank; Müller, Carsten; Wang, Le; Büchs, Jochen

2009-08-01

In industry and academic research, there is an increasing demand for flexible automated microfermentation platforms with advanced sensing technology. However, up to now, conventional platforms cannot generate continuous data in high-throughput cultivations, in particular for monitoring biomass and fluorescent proteins. Furthermore, microfermentation platforms are needed that can easily combine cost-effective, disposable microbioreactors with downstream processing and analytical assays. To meet this demand, a novel automated microfermentation platform consisting of a BioLector and a liquid-handling robot (Robo-Lector) was sucessfully built and tested. The BioLector provides a cultivation system that is able to permanently monitor microbial growth and the fluorescence of reporter proteins under defined conditions in microtiter plates. Three examplary methods were programed on the Robo-Lector platform to study in detail high-throughput cultivation processes and especially recombinant protein expression. The host/vector system E. coli BL21(DE3) pRhotHi-2-EcFbFP, expressing the fluorescence protein EcFbFP, was hereby investigated. With the method 'induction profiling' it was possible to conduct 96 different induction experiments (varying inducer concentrations from 0 to 1.5 mM IPTG at 8 different induction times) simultaneously in an automated way. The method 'biomass-specific induction' allowed to automatically induce cultures with different growth kinetics in a microtiter plate at the same biomass concentration, which resulted in a relative standard deviation of the EcFbFP production of only +/- 7%. The third method 'biomass-specific replication' enabled to generate equal initial biomass concentrations in main cultures from precultures with different growth kinetics. This was realized by automatically transferring an appropiate inoculum volume from the different preculture microtiter wells to respective wells of the main culture plate, where subsequently similar growth kinetics could be obtained. The Robo-Lector generates extensive kinetic data in high-throughput cultivations, particularly for biomass and fluorescence protein formation. Based on the non-invasive on-line-monitoring signals, actions of the liquid-handling robot can easily be triggered. This interaction between the robot and the BioLector (Robo-Lector) combines high-content data generation with systematic high-throughput experimentation in an automated fashion, offering new possibilities to study biological production systems. The presented platform uses a standard liquid-handling workstation with widespread automation possibilities. Thus, high-throughput cultivations can now be combined with small-scale downstream processing techniques and analytical assays. Ultimately, this novel versatile platform can accelerate and intensify research and development in the field of systems biology as well as modelling and bioprocess optimization.

High-resolution elemental mapping of human placental chorionic villi using synchrotron X-ray fluorescence spectroscopy

DOE PAGES

Punshon, Tracy; Chen, Si; Finney, Lydia; ...

2015-07-03

The placenta is the organ that mediates transport of nutrients and waste materials between mother and fetus. Synchrotron X-ray fluorescence (SXRF) microanalysis is a tool for imaging the distribution and quantity of elements in biological tissue, which can be used to study metal transport across biological membranes. Our aims were to pilot placental biopsy specimen preparation techniques that could be integrated into an ongoing epidemiology birth cohort study without harming rates of sample acquisition. We studied the effects of fixative (formalin or glutaraldehyde) and storage duration (30 days or immediate processing) on metal distribution and abundance and investigated a thaw-fixationmore » protocol for archived specimens stored at -80° C. We measured fixative elemental composition with and without a placental biopsy via inductively coupled plasma mass spectrometry (ICP-MS) to quantify fixative-induced elemental changes. Formalin-fixed specimens showed hemolysis of erythrocytes. The glutaraldehyde-paraformaldehyde solution in HEPES buffer (GTA-HEPES) had superior anatomical preservation, avoided hemolysis, and minimized elemental loss, although some cross-linking of exogenous Zn was evident. Elemental loss from tissue stored in fixative for 1 month showed variable losses (≈ 40 % with GTA-HEPES), suggesting storage duration be controlled for. Lastly, thawing of tissue held at -80 °C in a GTA-HEPES solution provided high-quality visual images and elemental images« less

Nerium oleander indirect leaf photosynthesis and light harvesting reductions after clipping injury or Spodoptera eridania herbivory: high sensitivity to injury.

PubMed

Delaney, Kevin J

2012-04-01

Variable indirect photosynthetic rate (P(n)) responses occur on injured leaves after insect herbivory. It is important to understand factors that influence indirect P(n) reductions after injury. The current study examines the relationship between gas exchange and chlorophyll a fluorescence parameters with injury intensity (% single leaf tissue removal) from clipping or Spodoptera eridania Stoll (Noctuidae) herbivory on Nerium oleander L. (Apocynaceae). Two experiments showed intercellular [CO(2)] increases but P(n) and stomatal conductance reductions with increasing injury intensity, suggesting non-stomatal P(n) limitation. Also, P(n) recovery was incomplete at 3d post-injury. This is the first report of a negative exponential P(n) impairment function with leaf injury intensity to suggest high N. oleander leaf sensitivity to indirect P(n) impairment. Negative linear functions occurred between most other gas exchange and chlorophyll a fluorescence parameters with injury intensity. The degree of light harvesting impairment increased with injury intensity via lower (1) photochemical efficiency indicated lower energy transfer efficiency from reaction centers to PSII, (2) photochemical quenching indicated reaction center closure, and (3) electron transport rates indicated less energy traveling through PSII. Future studies can examine additional mechanisms (mesophyll conductance, carbon fixation, and cardenolide induction) to cause N. oleander indirect leaf P(n) reductions after injury. Published by Elsevier Ireland Ltd.

75 FR 22031 - Energy Conservation Program for Consumer Products and Certain Commercial and Industrial Equipment...

Federal Register 2010, 2011, 2012, 2013, 2014

2010-04-27

..., including induction and fluorescent. Non-HID technologies are achieving market penetration in lighting... the electromagnetic (magnetic) type. The industry-accepted life of magnetic ballasts is 50,000 hours... analysis for today's proposed determination, DOE only considered magnetic ballasts because they are the...

A fluorescence-based method for rapid and direct determination of polybrominated diphenyl ethers in water

DOE PAGES

Shan, Huimei; Liu, Chongxuan; Wang, Zheming; ...

2015-01-01

A new method was developed for rapid and direct measurement of polybrominated diphenyl ethers (PBDEs) in aqueous samples using fluorescence spectroscopy. The fluorescence spectra of tri- to deca-BDE (BDE 28, 47, 99, 153, 190, and 209) commonly found in environment were measured at variable emission and excitation wavelengths. The results revealed that the PBDEs have distinct fluorescence spectral profiles and peak positions that can be exploited to identify these species and determine their concentrations in aqueous solutions. The detection limits as determined in deionized water spiked with PBDEs are 1.71-5.82 ng/L for BDE 28, BDE 47, BDE 190, and BDEmore » 209 and 45.55–69.95 ng/L for BDE 99 and BDE 153. The effects of environmental variables including pH, humic substance, and groundwater chemical composition on PBDEs measurements were also investigated. These environmental variables affected fluorescence intensity, but their effect can be corrected through linear additivity and separation of spectral signal contribution. Compared with conventional GC-based analytical methods, the fluorescence spectroscopy method is more efficient as it only uses a small amount of samples (2-4 mL), avoids lengthy complicated concentration and extraction steps, and has a low detection limit of a few ng/L.« less

Daily Course of CO2 Fluxes in the Atmosphere-Water System and Variable Fluorescence of Phytoplankton during the Open-Water Period for Lake Baikal according to Long-Term Measurements

NASA Astrophysics Data System (ADS)

Zavoruev, V. V.; Domysheva, V. M.; Pestunov, D. A.; Sakirko, M. V.; Panchenko, M. V.

2018-04-01

The process of gas exchange of CO2 in the atmosphere-water system and its relation to the daily course of variable fluorescence of phytoplankton is studied on the basis of long-term (2004-2014) measurements during the open water period for Lake Baikal. It is found that the decrease in photosynthetic activity of plankton is almost synchronous to the increase in the CO2 flux from atmosphere to water. It follows from comparison of the spring and summer data with December measurements that the daily decrease in variable fluorescence of phytoplankton is caused by the internal daily rhythm of the photosynthetic activity of plankton.

Assessment of variable fluorescence fluorometry as an approach for rapidly detecting living photoautotrophs in ballast water

NASA Astrophysics Data System (ADS)

First, Matthew R.; Robbins-Wamsley, Stephanie H.; Riley, Scott C.; Drake, Lisa A.

2018-03-01

Variable fluorescence fluorometry, an analytical approach that estimates the fluorescence yield of chlorophyll a (F0, a proximal measure of algal concentration) and photochemical yield (FV/FM, an indicator of the physiological status of algae) was evaluated as a means to rapidly assess photoautotrophs. Specifically, it was used to gauge the efficacy of ballast water treatment designed to reduce the transport and delivery of potentially invasive organisms. A phytoflagellate, Tetraselmis spp. (10-12 μm) and mixed communities of ambient protists were examined in both laboratory experiments and large-scale field trials simulating 5-d hold times in mock ballast tanks. In laboratory incubations, ambient organisms held in the dark exhibited declining F0 and FV/FM measurements relative to organisms held under lighted conditions. In field experiments, increases and decreases in F0 and FV/FM over the tank hold time corresponded to those of microscope counts of organisms in two of three trials. In the third trial, concentrations of organisms ≥ 10 and < 50 μm (presumably heterotrophic protists) increased while F0 and FV/FM decreased. Rapid and sensitive, variable fluorescence fluorometry is appropriate for detecting changes in organism concentrations and physiological status in samples dominated by microalgae. Changes in the heterotrophic community, which may become more prevalent in light-limited ballast tanks, would not be detected via variable fluorescence fluorometry, however.

Inductively coupled plasma atomic fluorescence spectrometric determination of cadmium, copper, iron, lead, manganese and zinc

USGS Publications Warehouse

Sanzolone, R.F.

1986-01-01

An inductively coupled plasma atomic fluorescence spectrometric method is described for the determination of six elements in a variety of geological materials. Sixteen reference materials are analysed by this technique to demonstrate its use in geochemical exploration. Samples are decomposed with nitric, hydrofluoric and hydrochloric acids, and the residue dissolved in hydrochloric acid and diluted to volume. The elements are determined in two groups based on compatibility of instrument operating conditions and consideration of crustal abundance levels. Cadmium, Cu, Pb and Zn are determined as a group in the 50-ml sample solution under one set of instrument conditions with the use of scatter correction. Limitations of the scatter correction technique used with the fluorescence instrument are discussed. Iron and Mn are determined together using another set of instrumental conditions on a 1-50 dilution of the sample solution without the use of scatter correction. The ranges of concentration (??g g-1) of these elements in the sample that can be determined are: Cd, 0.3-500; Cu, 0.4-500; Fe, 85-250 000; Mn, 45-100 000; Pb, 5-10 000; and Zn, 0.4-300. The precision of the method is usually less than 5% relative standard deviation (RSD) over a wide concentration range and acceptable accuracy is shown by the agreement between values obtained and those recommended for the reference materials.

Elemental ratios for characterization of quantum-dots populations in complex mixtures by asymmetrical flow field-flow fractionation on-line coupled to fluorescence and inductively coupled plasma mass spectrometry.

PubMed

Menendez-Miranda, Mario; Fernandez-Arguelles, Maria T; Costa-Fernandez, Jose M; Encinar, Jorge Ruiz; Sanz-Medel, Alfredo

2014-08-11

Separation and identification of nanoparticles of different composition, with similar particle diameter, coexisting in heterogeneous suspensions of polymer-coated CdSe/ZnS quantum dots (QDs) have been thoroughly assessed by asymmetric flow field-flow fractionation (AF4) coupled on-line to fluorescence and inductively coupled plasma mass spectrometry (ICPMS) detectors. Chemical characterization of any previously on-line separated nanosized species was achieved by the measurement of the elemental molar ratios of every element involved in the synthesis of the QDs, using inorganic standards and external calibration by flow injection analysis (FIA). Such elemental molar ratios, strongly limited so far to pure single nanoparticles suspensions, have been achieved with adequate accuracy by coupling for the first time an ICP-QQQ instrument to an AF4 system. This hyphenation turned out to be instrumental to assess the chemical composition of the different populations of nanoparticles coexisting in the relatively complex mixtures, due to its capabilities to detect the hardly detectable elements involved in the synthesis. Interestingly such information, complementary to that obtained by fluorescence, was very valuable to detect and identify unexpected nanosized species, present at significant level, produced during QDs synthesis and hardly detectable by standard approaches. Copyright © 2014. Published by Elsevier B.V.

Fluorescent magnetic hybrid nanoprobe for multimodal bioimaging

PubMed Central

Bright, Vanessa

2011-01-01

A fluorescent magnetic hybrid imaging nanoprobe (HINP) was fabricated by conjugation of superparamagnetic Fe3O4 nanoparticles and visible light-emitting (~600 nm) fluorescent CdTe/CdS quantum dots (QDs). The assembly strategy used the covalent linking of the oxidized dextran shell of magnetic particles to the glutathione ligands of QDs. Synthesized HINP formed stable water-soluble colloidal dispersions. The structure and properties of the particles were characterized by transmission electron and atomic force microscopy, energy dispersive X-ray analysis and inductively coupled plasma optical emission spectroscopy, dynamic light scattering analysis, optical absorption and photoluminescence spectroscopy, and fluorescent imaging. The luminescence imaging region of the nanoprobe was extended to the near-infrared (NIR) (~800 nm) by conjugation of superparamagnetic nanoparticles with synthesized CdHgTe/CdS QDs. Cadmium, mercury based QDs in HINP can be easily replaced by novel water soluble glutathione stabilized AgInS2/ZnS QDs to present a new class of cadmium-free multimodal imaging agents. Observed NIR photoluminescence of fluorescent magnetic nanocomposites supports their use for bioimaging. The developed HINP provides dual-imaging channels for simultaneous optical and magnetic resonance imaging. PMID:21597146

Validation of photosynthetic-fluorescence parameters as biomarkers for isoproturon toxic effect on alga Scenedesmus obliquus.

PubMed

Dewez, David; Didur, Olivier; Vincent-Héroux, Jonathan; Popovic, Radovan

2008-01-01

Photosynthetic-fluorescence parameters were investigated to be used as valid biomarkers of toxicity when alga Scenedesmus obliquus was exposed to isoproturon [3-(4-isopropylphenyl)-1,1-dimethylurea] effect. Chlorophyll fluorescence induction of algal cells treated with isoproturon showed inactivation of photosystem II (PSII) reaction centers and strong inhibition of PSII electron transport. A linear correlation was found (R2>or=0.861) between the change of cells density affected by isoproturon and the change of effective PSII quantum yield (PhiM'), photochemical quenching (qP) and relative photochemical quenching (qP(rel)) values. The cells density was also linearly dependent (R2=0.838) on the relative unquenched fluorescence parameter (UQF(rel)). Non-linear correlation was found (R2=0.937) only between cells density and the energy transfer efficiency from absorbed light to PSII reaction center (ABS/RC). The order of sensitivity determined by the EC-50% was: UQF(rel)>PhiM'>qP>qP(rel)>ABS/RC. Correlations between cells density and those photosynthetic-fluorescence parameters provide supporting evidence to use them as biomarkers of toxicity for environmental pollutants.

The effect of UV-B radiation on photosynthesis and respiration of phytoplankton, benthic macroalgae and seagrasses.

PubMed

Larkum, A W; Wood, W F

1993-04-01

Several species of marine benthic algae, four species of phytoplankton and two species of seagrass have been subjected to ultraviolet B irradiation for varying lengths of time and the effects on respiration, photosynthesis and fluorescence rise kinetics studied. No effect on respiration was found. Photosynthesis was inhibited to a variable degree in all groups of plants after irradiation over periods of up to 1 h and variable fluorescence was also inhibited in a similar way. The most sensitive plants were phytoplankton and deep-water benthic algae. Intertidal benthic algae were the least sensitive to UV-B irradiation and this may be related to adaptation, through the accumulation of UV-B screening compounds, to high light/high UV-B levels. Inhibition of variable fluorescence (Fv) of the fluorescence rise curve was a fast and sensitive indicator of UV-B damage. Two plants studied, a brown alga and a seagrass, showed very poor recovery of Fv over a period of 32 h.

In vivo analysis of intestinal permeability following hemorrhagic shock

PubMed Central

Alsaigh, Tom; Chang, Marisol; Richter, Michael; Mazor, Rafi; Kistler, Erik B

2015-01-01

AIM: To determine the time course of intestinal permeability changes to proteolytically-derived bowel peptides in experimental hemorrhagic shock. METHODS: We injected fluorescently-conjugated casein protein into the small bowel of anesthetized Wistar rats prior to induction of experimental hemorrhagic shock. These molecules, which fluoresce when proteolytically cleaved, were used as markers for the ability of proteolytically cleaved intestinal products to access the central circulation. Blood was serially sampled to quantify the relative change in concentration of proteolytically-cleaved particles in the systemic circulation. To provide spatial resolution of their location, particles in the mesenteric microvasculature were imaged using in vivo intravital fluorescent microscopy. The experiments were then repeated using an alternate measurement technique, fluorescein isothiocyanate (FITC)-labeled dextrans 20, to semi-quantitatively verify the ability of bowel-derived low-molecular weight molecules (< 20 kD) to access the central circulation. RESULTS: Results demonstrate a significant increase in systemic permeability to gut-derived peptides within 20 min after induction of hemorrhage (1.11 ± 0.19 vs 0.86 ± 0.07, P < 0.05) compared to control animals. Reperfusion resulted in a second, sustained increase in systemic permeability to gut-derived peptides in hemorrhaged animals compared to controls (1.2 ± 0.18 vs 0.97 ± 0.1, P < 0.05). Intravital microscopy of the mesentery also showed marked accumulation of fluorescent particles in the microcirculation of hemorrhaged animals compared to controls. These results were replicated using FITC dextrans 20 [10.85 ± 6.52 vs 3.38 ± 1.11 fluorescent intensity units (× 105, P < 0.05, hemorrhagic shock vs controls)], confirming that small bowel ischemia in response to experimental hemorrhagic shock results in marked and early increases in gut membrane permeability. CONCLUSION: Increased small bowel permeability in hemorrhagic shock may allow for systemic absorption of otherwise retained proteolytically-generated peptides, with consequent hemodynamic instability and remote organ failure. PMID:26557479

Non-invasive In Vivo Fluorescence Optical Imaging of Inflammatory MMP Activity Using an Activatable Fluorescent Imaging Agent.

PubMed

Schwenck, Johannes; Maier, Florian C; Kneilling, Manfred; Wiehr, Stefan; Fuchs, Kerstin

2017-05-08

This paper describes a non-invasive method for imaging matrix metalloproteinases (MMP)-activity by an activatable fluorescent probe, via in vivo fluorescence optical imaging (OI), in two different mouse models of inflammation: a rheumatoid arthritis (RA) and a contact hypersensitivity reaction (CHR) model. Light with a wavelength in the near infrared (NIR) window (650 - 950 nm) allows a deeper tissue penetration and minimal signal absorption compared to wavelengths below 650 nm. The major advantages using fluorescence OI is that it is cheap, fast and easy to implement in different animal models. Activatable fluorescent probes are optically silent in their inactivated states, but become highly fluorescent when activated by a protease. Activated MMPs lead to tissue destruction and play an important role for disease progression in delayed-type hypersensitivity reactions (DTHRs) such as RA and CHR. Furthermore, MMPs are the key proteases for cartilage and bone degradation and are induced by macrophages, fibroblasts and chondrocytes in response to pro-inflammatory cytokines. Here we use a probe that is activated by the key MMPs like MMP-2, -3, -9 and -13 and describe an imaging protocol for near infrared fluorescence OI of MMP activity in RA and control mice 6 days after disease induction as well as in mice with acute (1x challenge) and chronic (5x challenge) CHR on the right ear compared to healthy ears.

Application of fluorescence spectroscopy for on-line bioprocess monitoring and control

NASA Astrophysics Data System (ADS)

Boehl, Daniela; Solle, D.; Toussaint, Hans J.; Menge, M.; Renemann, G.; Lindemann, Carsten; Hitzmann, Bernd; Scheper, Thomas-Helmut

2001-02-01

12 Modern bioprocess control requires fast data acquisition and in-time evaluation of bioprocess variables. On-line fluorescence spectroscopy for data acquisition and the use of chemometric methods accomplish these requirements. The presented investigations were performed with fluorescence spectrometers with wide ranges of excitation and emission wavelength. By detection of several biogenic fluorophors (amino acids, coenzymes and vitamins) a large amount of information about the state of the bioprocess are obtained. For the evaluation of the process variables partial least squares regression is used. This technique was applied to several bioprocesses: the production of ergotamine by Claviceps purpurea, the production of t-PA (tissue plasminogen activator) by animal cells and brewing processes. The main point of monitoring the brewing processes was to determine the process variables cell count and extract concentration.

Acclimation of shade-tolerant and light-resistant Tradescantia species to growth light: chlorophyll a fluorescence, electron transport, and xanthophyll content.

PubMed

Mishanin, Vladimir I; Trubitsin, Boris V; Patsaeva, Svetlana V; Ptushenko, Vasily V; Solovchenko, Alexei E; Tikhonov, Alexander N

2017-09-01

In this study, we have compared the photosynthetic characteristics of two contrasting species of Tradescantia plants, T. fluminensis (shade-tolerant species), and T. sillamontana (light-resistant species), grown under the low light (LL, 50-125 µmol photons m -2  s -1 ) or high light (HL, 875-1000 µmol photons m -2  s -1 ) conditions during their entire growth period. For monitoring the functional state of photosynthetic apparatus (PSA), we measured chlorophyll (Chl) a emission fluorescence spectra and kinetics of light-induced changes in the heights of fluorescence peaks at 685 and 740 nm (F 685 and F 740 ). We also compared the light-induced oxidation of P 700 and assayed the composition of carotenoids in Tradescantia leaves grown under the LL and HL conditions. The analyses of slow induction of Chl a fluorescence (SIF) uncovered different traits in the LL- and HL-grown plants of ecologically contrasting Tradescantia species, which may have potential ecophysiological significance with respect to their tolerance to HL stress. The fluorometry and EPR studies of induction events in chloroplasts in situ demonstrated that acclimation of both Tradescantia species to HL conditions promoted faster responses of their PSA as compared to LL-grown plants. Acclimation of both species to HL also caused marked changes in the leaf anatomy and carotenoid composition (an increase in Violaxanthin + Antheraxantin + Zeaxanthin and Lutein pools), suggesting enhanced photoprotective capacity of the carotenoids in the plants grown in nature under high irradiance. Collectively, the results of the present work suggest that the mechanisms of long-term PSA photoprotection in Tradescantia are based predominantly on the light-induced remodeling of pigment-protein complexes in chloroplasts.

Investigation of self-excited induction generators for wind turbine applications

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

Muljadi, E.; Butterfield, C.P.; Sallan, J.

2000-02-28

The use of squirrel-cage induction machines in wind generation is widely accepted as a generator of choice. The squirrel-cage induction machine is simple, reliable, cheap, lightweight, and requires very little maintenance. Generally, the induction generator is connected to the utility at constant frequency. With a constant frequency operation, the induction generator operates at practically constant speed (small range of slip). The wind turbine operates in optimum efficiency only within a small range of wind speed variation. The variable-speed operation allows an increase in energy captured and reduces both the torque peaks in the drive train and the power fluctuations sentmore » to the utility. In variable-speed operation, an induction generator needs an interface to convert the variable frequency output of the generator to the fixed frequency at the utility. This interface can be simplified by using a self-excited generator because a simple diode bridge is required to perform the ac/dc conversion. The subsequent dc/ac conversion can be performed using different techniques. The use of a thyristor bridge is readily available for large power conversion and has a lower cost and higher reliability. The firing angle of the inverter bridge can be controlled to track the optimum power curve of the wind turbine. With only diodes and thyristors used in power conversion, the system can be scaled up to a very high voltage and high power applications. This paper analyzes the operation of such a system applied to a 1/3-hp self-excited induction generator. It includes the simulations and tests performed for the different excitation configurations.« less

DNA-encapsulated magnesium phosphate nanoparticles elicit both humoral and cellular immune responses in mice

PubMed Central

Bhakta, Gajadhar; Nurcombe, Victor; Maitra, Amarnath; Shrivastava, Anju

2014-01-01

The efficacy of pEGFP (plasmid expressing enhanced green fluorescent protein)-encapsulated PEGylated (meaning polyethylene glycol coated) magnesium phosphate nanoparticles (referred to as MgPi-pEGFP nanoparticles) for the induction of immune responses was investigated in a mouse model. MgPi-pEGFP nanoparticles induced enhanced serum antibody and antigen-specific T-lymphocyte responses, as well as increased IFN-? and IL-12 levels compared to naked pEGFP when administered via intravenous, intraperitoneal or intramuscular routes. A significant macrophage response, both in size and activity, was also observed when mice were immunized with the nanoparticle formulation. The response was highly specific for the antigen, as the increase in interaction between macrophages and lymphocytes as well as lymphocyte proliferation took place only when they were re-stimulated with recombinant green fluorescence protein (rGFP). Thus the nanoparticle formulation elicited both humoral as well as cellular responses. Cytokine profiling revealed the induction of Th-1 type responses. The results suggest DNA-encapsulated magnesium phosphate (MgPi) nanoparticles may constitute a safer, more stable and cost-efficient DNA vaccine formulation. PMID:24936399

Correlating Fluorescence and High-Resolution Scanning Electron Microscopy (HRSEM) for the study of GABAA receptor clustering induced by inhibitory synaptic plasticity.

PubMed

Orlando, Marta; Ravasenga, Tiziana; Petrini, Enrica Maria; Falqui, Andrea; Marotta, Roberto; Barberis, Andrea

2017-10-23

Both excitatory and inhibitory synaptic contacts display activity dependent dynamic changes in their efficacy that are globally termed synaptic plasticity. Although the molecular mechanisms underlying glutamatergic synaptic plasticity have been extensively investigated and described, those responsible for inhibitory synaptic plasticity are only beginning to be unveiled. In this framework, the ultrastructural changes of the inhibitory synapses during plasticity have been poorly investigated. Here we combined confocal fluorescence microscopy (CFM) with high resolution scanning electron microscopy (HRSEM) to characterize the fine structural rearrangements of post-synaptic GABA A Receptors (GABA A Rs) at the nanometric scale during the induction of inhibitory long-term potentiation (iLTP). Additional electron tomography (ET) experiments on immunolabelled hippocampal neurons allowed the visualization of synaptic contacts and confirmed the reorganization of post-synaptic GABA A R clusters in response to chemical iLTP inducing protocol. Altogether, these approaches revealed that, following the induction of inhibitory synaptic potentiation, GABA A R clusters increase in size and number at the post-synaptic membrane with no other major structural changes of the pre- and post-synaptic elements.

A flow cytometry-optimized assay using an SOS-green fluorescent protein (SOS-GFP) whole-cell biosensor for the detection of genotoxins in complex environments.

PubMed

Norman, Anders; Hansen, Lars Hestbjerg; Sørensen, Søren J

2006-02-28

Whole-cell biosensors have become popular tools for detection of ecotoxic compounds in environmental samples. We have developed an assay optimized for flow cytometry with detection of genotoxic compounds in mind. The assay features extended pre-incubation and a cell density of only 10(6)-10(7) cells/mL, and proved far more sensitive than a previously published assay using the same biosensor strain. By applying the SOS-green fluorescent protein (GFP) whole-cell biosensor directly to soil microcosms we were also able to evaluate both the applicability and sensitivity of a biosensor based on SOS-induction in whole soil samples. Soil microcosms were spiked with a dilution-series of crude broth extract from the mitomycin C-producing streptomycete Streptomyces caespitosus. Biosensors extracted from these microcosms after 1 day of incubation at 30 degrees C were easily distinguished from extracts of non-contaminated soil particles when using flow cytometry, and induction of the biosensor by mitomycin C was detectable at concentrations as low as 2.5 ng/g of soil.

Disruption of the ndhF1 gene affects Chl fluorescence through state transition in the Cyanobacterium Synechocystis sp. PCC 6803, resulting in apparent high efficiency of photosynthesis.

PubMed

Ogawa, Takako; Harada, Tetsuyuki; Ozaki, Hiroshi; Sonoike, Kintake

2013-07-01

In Synechocystis sp. PCC 6803, the disruption of the ndhF1 gene (slr0844), which encodes a subunit of one of the NDH-1 complexes (NDH-1L complex) serving for respiratory electron transfer, causes the largest change in Chl fluorescence induction kinetics among the kinetics of 750 disruptants searched in the Fluorome, the cyanobacterial Chl fluorescence database. The cause of the explicit phenotype of the ndhF1 disruptant was examined by measurements of the photosynthetic rate, Chl fluorescence and state transition. The results demonstrate that the defects in respiratory electron transfer obviously have great impact on Chl fluorescence in cyanobacteria. The inactivation of NDH-1L complexes involving electron transfer from NDH-1 to plastoquinone (PQ) would result in the oxidation of the PQ pool, leading to the transition to State 1, where the yield of Chl fluorescence is high. Apparently, respiration, although its rate is far lower than that of photosynthesis, could affect Chl fluorescence through the state transition as leverage. The disruption of the ndhF1 gene caused lower oxygen-evolving activity but the estimated electron transport rate from Chl fluorescence measurements was faster in the mutant than in the wild-type cells. The discrepancy could be ascribed to the decreased level of non-photochemical quenching due to state transition. One must be cautious when using the Chl fluorescence parameter to estimate photosynthesis in mutants defective in state transition.

Portable total reflection x-ray fluorescence analysis in the identification of unknown laboratory hazards

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

Liu, Ying, E-mail: liu.ying.48r@st.kyoto-u.ac.jp; Imashuku, Susumu; Sasaki, Nobuharu

In this study, a portable total reflection x-ray fluorescence (TXRF) spectrometer was used to analyze unknown laboratory hazards that precipitated on exterior surfaces of cooling pipes and fume hood pipes in chemical laboratories. With the aim to examine the accuracy of TXRF analysis for the determination of elemental composition, analytical results were compared with those of wavelength-dispersive x-ray fluorescence spectrometry, scanning electron microscope and energy-dispersive x-ray spectrometry, energy-dispersive x-ray fluorescence spectrometry, inductively coupled plasma atomic emission spectrometry, x-ray diffraction spectrometry (XRD), and x-ray photoelectron spectroscopy (XPS). Detailed comparison of data confirmed that the TXRF method itself was not sufficient tomore » determine all the elements (Z > 11) contained in the samples. In addition, results suggest that XRD should be combined with XPS in order to accurately determine compound composition. This study demonstrates that at least two analytical methods should be used in order to analyze the composition of unknown real samples.« less

Ca2+ mobilization during cell death induction by sodium 5, 6-benzylidene-L-ascorbate.

PubMed

Takahashi, H; Sakagami, H; Ohata, H; Iida, M; Momose, K; Yamamura, M; Takeda, M

1996-01-01

The cytotoxic activity of sodium 5,6-benzylidene-L-ascorbate (SBA) against human KG-1-C glioma and T98G glioblastoma cell lines was augmented by pretreatment of the cells with L-buthionine-[S, R]-sulfoximine (BSO), which reduced the intracellular glutathione concentrations. SBA produced shrunken cells and large DNA fragments, without the induction of nuclear and internucleosomal DNA fragmentation. The rapid elevation of intracellular free Ca2+ concentration observed after SBA treatment was further augmented by BSO pretreatment. A confocal experiment with Fluo-3 fluorescence revealed that SBA markedly elevated the free Ca2+ concentration in the nuclear region, but did not significantly affect that in the cytoplasmic region. The present study suggests that the nuclear accumulation of Ca2+ is an important initial step for cell death induction by SBA.

Phytoplankton natural fluorescence variability in the Sargasso Sea

NASA Astrophysics Data System (ADS)

Westberry, T. K.; Siegel, D. A.

2003-03-01

Phytoplankton fluorescence has been used historically as a means of assessing phytoplankton biomass, rates of primary production (PP) and physiological status in laboratory, in situ, and satellite based investigations. Assumptions about the quantum yield of phytoplankton fluorescence, φf, are often overlooked and can become problematic when fluorescence based methods are applied. A time series of φf observations from the northwestern Sargasso Sea is presented with the goal of understanding the controls on fluorescence and its applicability for assessing upper ocean biological processes. Accurate estimates of φf require accounting for Raman scattering and the conversion of planar to scalar irradiance. Variability in φf occurs on both seasonal and episodic time scales. Seasonal variations show maxima in the surface layer during summer months while lower, more uniform values are found throughout the winter when deep mixing occurs. Large episodic variations in φf are observed throughout the record which dwarf seasonal changes. Predictions of depth-dependent and depth-integrated PP rates using φf and natural fluorescence fluxes are only marginally successful ( r2˜50%), although comparable with results from global bio-optical models for the Sargasso Sea. Improvements in PP predictions are hindered by weak statistical relationships with other parameters. φf is largely decoupled from the quantum yield of carbon assimilation, φc, indicating that an inverse relationship between fluorescence and photosynthesis does not exist. Consequently, variability in the quantum yield of thermal de-excitation, φh, is found to be of similar magnitude as φf on the timescales observed. These observations show that assumptions about photochemical energy flow through the phytoplankton community must be made carefully and that the fluorescence-photosynthesis relationship is not straightforward.

Drivers and variability of the Chl fluorescence emission spectrum from the leaf through the canopy

NASA Astrophysics Data System (ADS)

Magney, T. S.; Frankenberg, C.; Grossman, K.; Koehler, P.; North, G.; Porcar-Castell, A.; Stutz, J.; Fisher, J.

2017-12-01

Recent advances in the retrieval of solar induced chlorophyll fluorescence (SIF) from remote sensing platforms provide a significant step towards mapping instantaneous plant photosynthesis across space and time. However, our current understanding of the variability and controls on the shape of the chlorophyll fluorescence (ChlF) spectrum is limited. To address these uncertainties, we have developed instrumentation to make highly resolved spectral measurements of SIF from both leaf and canopy scales. At the leaf scale, we simultaneously collected active (PAM) and passive (675-850 nm) fluorescence with photosynthesis across a range of species and conditions; and at the canopy scale, diurnal and seasonal Fraunhofer-based SIF retrievals across the red and far-red spectrum are made at four different flux tower sites (Costa Rica, Iowa (2), and Colorado). From both of these scales we are able to determine (1) the variability in steady-state spectra across species and individuals; and (2) the environmental, functional, and structural controls on SIF. Here we report on the sensitivity of SIF spectra from a singular value decomposition analysis; and present on the mechanisms - pigment concentration, species, non-photochemical and photochemical quenching, and environmental conditions - controlling SIF variability. Further, we will discuss how an improved understanding of leaf-level variability can inform canopy level SIF, and ultimately how such information may enable proper interpretation of satellite retrievals.

Optical phantoms with variable properties and geometries for diffuse and fluorescence optical spectroscopy

NASA Astrophysics Data System (ADS)

Leh, Barbara; Siebert, Rainer; Hamzeh, Hussein; Menard, Laurent; Duval, Marie-Alix; Charon, Yves; Abi Haidar, Darine

2012-10-01

Growing interest in optical instruments for biomedical applications has increased the use of optically calibrated phantoms. Often associated with tissue modeling, phantoms allow the characterization of optical devices for clinical purposes. Fluorescent gel phantoms have been developed, mimicking optical properties of healthy and tumorous brain tissues. Specific geometries of dedicated molds offer multiple-layer phantoms with variable thicknesses and monolayer phantoms with cylindrical inclusions at various depths and diameters. Organic chromophores are added to allow fluorescence spectroscopy. These phantoms are designed to be used with 405 nm as the excitation wavelength. This wavelength is then adapted to excite large endogenous molecules. The benefits of these phantoms in understanding fluorescence tissue analysis are then demonstrated. In particular, detectability aspects as a function of geometrical and optical parameters are presented and discussed.

Laser fluorescence fluctuation excesses in molecular immunology experiments

NASA Astrophysics Data System (ADS)

Galich, N. E.; Filatov, M. V.

2007-04-01

A novel approach to statistical analysis of flow cytometry fluorescence data have been developed and applied for population analysis of blood neutrophils stained with hydroethidine during respiratory burst reaction. The staining based on intracellular oxidation hydroethidine to ethidium bromide, which intercalate into cell DNA. Fluorescence of the resultant product serves as a measure of the neutrophil ability to generate superoxide radicals after induction respiratory burst reaction by phorbol myristate acetate (PMA). It was demonstrated that polymorphonuclear leukocytes of persons with inflammatory diseases showed a considerably changed response. Cytofluorometric histograms obtained have unique information about condition of neutrophil population what might to allow a determination of the pathology processes type connecting with such inflammation. A novel approach to histogram analysis is based on analysis of high-momentum dynamic of distribution. The features of fluctuation excesses of distribution have unique information about disease under consideration.

Thylakoid membrane model of the Chl a fluorescence transient and P700 induction kinetics in plant leaves.

PubMed

Belyaeva, N E; Bulychev, A A; Riznichenko, G Yu; Rubin, A B

2016-12-01

A new Thylakoid model is presented, which describes in detail the electron/proton transfer reactions between membrane protein complexes including photosystems II and I (PSII, PSI), cytochrome (Cyt) b 6 f, mobile plastoquinone PQ pool in the thylakoid membrane, plastocyanin in lumen and ferredoxin in stroma, reduction of NADP via FNR and cyclic electron transfer. The Thylakoid model parameters were fitted both to Chl fluorescence induction data (FI) and oxido-reductions of P700 (ΔA 810 ) measured from 20 μs up to 20 s in pea leaves. The two-wave kinetics of FI and ΔA 810 (O(JI)PSM and OABCDE) were described quantitatively, provided that the values of membrane electrochemical potential components ΔΨ(t), pH L (t)/pH S (t) are in physiologically relevant ranges. The time courses on the time scale from nanoseconds to tens of seconds of oxido-reduction changes of ET components as well as concentrations of proton/ions (K + , Cl - ) were calculated. We assume a low constant FNR activity over this period. Charge movements across the thylakoid membrane by passive leakage and active ATPase transport and proton buffer reactions are simulated. The dynamics of charge fluxes during photosynthetic induction under low light (PFD 200 μmol photons m -2  s -1 ) were analyzed. The initial wave of P700 oxidation within 20 ms during independent operation of PSI and PSII was followed after 50 ms by PSI donor-side reduction from reduced PQ pool via Cyt b 6 f site. The Cyt b 6 f reactions contribute to the stabilization of fluxes in the time range 1 s < t < 10 s. The detailed analysis of Chl a fluorescence at the PSM stage (t > 10 s) would need the investigation of FNR activation effect in order to explain the transitions between cyclic and linear electron transport.

Study of the Many Fluorescent Lines and the Absorption Variability in GX 301-2 with XMM-Newton

NASA Technical Reports Server (NTRS)

Fuerst, F.; Suchy, S.; Kreykenbohm, I.; Barragan, L.; Wilms, J.; Pottschmidt, K.; Caballero, I.; Kretschmar, P.; Ferrigno, C.; Rothschild, R. E.

2011-01-01

We present an in-depth study of the High Mass X-ray Binary (HMXB) GX 301-2 during its pre-periastron flare using data from the XMM-Newton satellite. The energy spectrum shows a power law continuum absorbed by a large equivalent hydrogen column on the order of 10(exp 24)/ sq cm and a prominent Fe K-alpha fluorescent emission line. Besides the Fe K-alpha line, evidence for Fe K-Beta, Ni K-alpha, Ni K-Beta, S K-alpha, Ar K-alpha, Ca K-alpha, and Cr K-alpha fluorescent lines is found. The observed line strengths are consistent with fluorescence in a cold absorber. This is the first time that Cr K-alpha is seen in emission in the X-ray spectrum of a HMXB. In addition to the modulation by the strong pulse period of approx 685 sec the source is highly variable and shows different states of activity. We perform time-resolved as well as pulse-to-pulse resolved spectroscopy to investigate differences between these states of activity. We find that fluorescent line fluxes are strongly variable and generally follow the overall flux. The N-H value is variable by a factor of 2, but not correlated to continuum normalization. We find an interval of low flux in the light curve in which the pulsations cease almost completely, without any indication of an increasing absorption column. We investigate this dip in detail and argue that it is most likely that during the dip the accretion ceased and the afterglow of the fluorescent iron accounted for the main portion of the X-ray flux. A similar dip was found earlier in RXTE data, and we compare our findings to these results.

Long-working-distance fluorescence microscope with high-numerical-aperture objectives for variable-magnification imaging in live mice from macro- to subcellular

NASA Astrophysics Data System (ADS)

Kimura, Hiroaki; Momiyama, Masashi; Tomita, Katsuro; Tsuchiya, Hiroyuki; Hoffman, Robert M.

2010-11-01

We demonstrate the development of a long-working-distance fluorescence microscope with high-numerical-aperture objectives for variable-magnification imaging in live mice from macro- to subcellular. To observe cytoplasmic and nuclear dynamics of cancer cells in the living mouse, 143B human osteosarcoma cells are labeled with green fluorescent protein in the nucleus and red fluorescent protein in the cytoplasm. These dual-color cells are injected by a vascular route in an abdominal skin flap in nude mice. The mice are then imaged with the Olympus MVX10 macroview fluorescence microscope. With the MVX10, the nuclear and cytoplasmic behavior of cancer cells trafficking in blood vessels of live mice is observed. We also image lung metastases in live mice from the macro- to the subcellular level by opening the chest wall and imaging the exposed lung in live mice. Injected splenocytes, expressing cyan fluorescent protein, could also be imaged on the lung of live mice. We demonstrate that the MVX10 microscope offers the possibility of full-range in vivo fluorescence imaging from macro- to subcellular and should enable widespread use of powerful imaging technologies enabled by genetic reporters and other fluorophores.

Robo-Lector – a novel platform for automated high-throughput cultivations in microtiter plates with high information content

PubMed Central

Huber, Robert; Ritter, Daniel; Hering, Till; Hillmer, Anne-Kathrin; Kensy, Frank; Müller, Carsten; Wang, Le; Büchs, Jochen

2009-01-01

Background In industry and academic research, there is an increasing demand for flexible automated microfermentation platforms with advanced sensing technology. However, up to now, conventional platforms cannot generate continuous data in high-throughput cultivations, in particular for monitoring biomass and fluorescent proteins. Furthermore, microfermentation platforms are needed that can easily combine cost-effective, disposable microbioreactors with downstream processing and analytical assays. Results To meet this demand, a novel automated microfermentation platform consisting of a BioLector and a liquid-handling robot (Robo-Lector) was sucessfully built and tested. The BioLector provides a cultivation system that is able to permanently monitor microbial growth and the fluorescence of reporter proteins under defined conditions in microtiter plates. Three examplary methods were programed on the Robo-Lector platform to study in detail high-throughput cultivation processes and especially recombinant protein expression. The host/vector system E. coli BL21(DE3) pRhotHi-2-EcFbFP, expressing the fluorescence protein EcFbFP, was hereby investigated. With the method 'induction profiling' it was possible to conduct 96 different induction experiments (varying inducer concentrations from 0 to 1.5 mM IPTG at 8 different induction times) simultaneously in an automated way. The method 'biomass-specific induction' allowed to automatically induce cultures with different growth kinetics in a microtiter plate at the same biomass concentration, which resulted in a relative standard deviation of the EcFbFP production of only ± 7%. The third method 'biomass-specific replication' enabled to generate equal initial biomass concentrations in main cultures from precultures with different growth kinetics. This was realized by automatically transferring an appropiate inoculum volume from the different preculture microtiter wells to respective wells of the main culture plate, where subsequently similar growth kinetics could be obtained. Conclusion The Robo-Lector generates extensive kinetic data in high-throughput cultivations, particularly for biomass and fluorescence protein formation. Based on the non-invasive on-line-monitoring signals, actions of the liquid-handling robot can easily be triggered. This interaction between the robot and the BioLector (Robo-Lector) combines high-content data generation with systematic high-throughput experimentation in an automated fashion, offering new possibilities to study biological production systems. The presented platform uses a standard liquid-handling workstation with widespread automation possibilities. Thus, high-throughput cultivations can now be combined with small-scale downstream processing techniques and analytical assays. Ultimately, this novel versatile platform can accelerate and intensify research and development in the field of systems biology as well as modelling and bioprocess optimization. PMID:19646274

Investigating Electromagnetic Induction through a Microcomputer-Based Laboratory.

ERIC Educational Resources Information Center

Trumper, Ricardo; Gelbman, Moshe

2000-01-01

Describes a microcomputer-based laboratory experiment designed for high school students that very accurately analyzes Faraday's law of electromagnetic induction, addressing each variable separately while the others are kept constant. (Author/CCM)

Reassessing the Role of Type II Toxin-Antitoxin Systems in Formation of Escherichia coli Type II Persister Cells.

PubMed

Goormaghtigh, Frédéric; Fraikin, Nathan; Putrinš, Marta; Hallaert, Thibaut; Hauryliuk, Vasili; Garcia-Pino, Abel; Sjödin, Andreas; Kasvandik, Sergo; Udekwu, Klas; Tenson, Tanel; Kaldalu, Niilo; Van Melderen, Laurence

2018-06-12

Persistence is a reversible and low-frequency phenomenon allowing a subpopulation of a clonal bacterial population to survive antibiotic treatments. Upon removal of the antibiotic, persister cells resume growth and give rise to viable progeny. Type II toxin-antitoxin (TA) systems were assumed to play a key role in the formation of persister cells in Escherichia coli based on the observation that successive deletions of TA systems decreased persistence frequency. In addition, the model proposed that stochastic fluctuations of (p)ppGpp levels are the basis for triggering activation of TA systems. Cells in which TA systems are activated are thought to enter a dormancy state and therefore survive the antibiotic treatment. Using independently constructed strains and newly designed fluorescent reporters, we reassessed the roles of TA modules in persistence both at the population and single-cell levels. Our data confirm that the deletion of 10 TA systems does not affect persistence to ofloxacin or ampicillin. Moreover, microfluidic experiments performed with a strain reporting the induction of the yefM-yoeB TA system allowed the observation of a small number of type II persister cells that resume growth after removal of ampicillin. However, we were unable to establish a correlation between high fluorescence and persistence, since the fluorescence of persister cells was comparable to that of the bulk of the population and none of the cells showing high fluorescence were able to resume growth upon removal of the antibiotic. Altogether, these data show that there is no direct link between induction of TA systems and persistence to antibiotics. IMPORTANCE Within a growing bacterial population, a small subpopulation of cells is able to survive antibiotic treatment by entering a transient state of dormancy referred to as persistence. Persistence is thought to be the cause of relapsing bacterial infections and is a major public health concern. Type II toxin-antitoxin systems are small modules composed of a toxic protein and an antitoxin protein counteracting the toxin activity. These systems were thought to be pivotal players in persistence until recent developments in the field. Our results demonstrate that previous influential reports had technical flaws and that there is no direct link between induction of TA systems and persistence to antibiotics. Copyright © 2018 Goormaghtigh et al.

Detection of a variable number of ribosomal DNA loci by fluorescent in situ hybridization in Populus species.

PubMed

Prado, E A; Faivre-Rampant, P; Schneider, C; Darmency, M A

1996-10-01

Fluorescent in situ hybridization (FISH) was applied to related Populus species (2n = 19) in order to detect rDNA loci. An interspecific variability in the number of hybridization sites was revealed using as probe an homologous 25S clone from Populus deltoides. The application of image analysis methods to measure fluorescence intensity of the hybridization signals has enabled us to characterize major and minor loci in the 18S-5.8S-25S rDNA. We identified one pair of such rDNA clusters in Populus alba; two pairs, one major and one minor, in both Populus nigra and P. deltoides; and three pairs in Populus balsamifera, (two major and one minor) and Populus euroamericana (one major and two minor). FISH results are in agreement with those based on RFLP analysis. The pBG13 probe containing 5S sequence from flax detected two separate clusters corresponding to the two size classes of units that coexist within 5S rDNA of most Populus species. Key words : Populus spp., fluorescent in situ hybridization, FISH, rDNA variability, image analysis.

Absorption and fluorescence properties of colored dissolved organic matter in the Ross Sea during austral summer

NASA Astrophysics Data System (ADS)

D'Sa, E. J.; Kim, H. C.; Ha, S. Y.

2016-12-01

Colored dissolved organic matter (CDOM) spectral absorption and excitation-emission matrix (EEMs) fluorescence with parallel factor analysis (PARAFAC) were examined in the Ross Sea during a survey conducted on board the R/V Araon in the austral summer of 14/15. CDOM absorption at 355 nm ranged from 0.06 to 1.14 m-1 while spectral slope S calculated between 275-295 nm wavelength ranged from 18.83 to 33.32 µm-1 with water masses playing an important role in its variability. Spectral slope S decreased with increasing CDOM absorption indicating the strong role of photo-oxidation on CDOM abundance during the summer. PARAFAC analysis of EEM data identified two humic-like (terrestrial and marine-like) and a protein-like (tryptophan-like) component. The two humic-like components were well correlated with little variability spatially and across the water column ( 0-100 m) likely indicating more refractory material. The protein-like fluorescent component was relatively quite variable supporting the autochthonous production of this fluorescent component in the highly productive Ross Sea waters.

Fluorescence-encoded gold nanoparticles: library design and modulation of cellular uptake into dendritic cells.

PubMed

Rodriguez-Lorenzo, Laura; Fytianos, Kleanthis; Blank, Fabian; von Garnier, Christophe; Rothen-Rutishauser, Barbara; Petri-Fink, Alke

2014-04-09

In order to harness the unique properties of nanoparticles for novel clinical applications and to modulate their uptake into specific immune cells we designed a new library of homo- and hetero-functional fluorescence-encoded gold nanoparticles (Au-NPs) using different poly(vinyl alcohol) and poly(ethylene glycol)-based polymers for particle coating and stabilization. The encoded particles were fully characterized by UV-Vis and fluorescence spectroscopy, zeta potential and dynamic light scattering. The uptake by human monocyte derived dendritic cells in vitro was studied by confocal laser scanning microscopy and quantified by fluorescence-activated cell sorting and inductively coupled plasma atomic emission spectroscopy. We show how the chemical modification of particle surfaces, for instance by attaching fluorescent dyes, can conceal fundamental particle properties and modulate cellular uptake. In order to mask the influence of fluorescent dyes on cellular uptake while still exploiting its fluorescence for detection, we have created hetero-functionalized Au-NPs, which again show typical particle dependent cellular interactions. Our study clearly prove that the thorough characterization of nanoparticles at each modification step in the engineering process is absolutely essential and that it can be necessary to make substantial adjustments of the particles in order to obtain reliable cellular uptake data, which truly reflects particle properties. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Novel fluorescence molecular imaging of chemotherapy-induced intestinal apoptosis

NASA Astrophysics Data System (ADS)

Levin, Galit; Shirvan, Anat; Grimberg, Hagit; Reshef, Ayelet; Yogev-Falach, Merav; Cohen, Avi; Ziv, Ilan

2009-09-01

Chemotherapy-induced enteropathy (CIE) is one of the most serious complications of anticancer therapy, and tools for its early detection and monitoring are highly needed. We report on a novel fluorescence method for detection of CIE, based on molecular imaging of the related apoptotic process. The method comprises systemic intravenous administration of the ApoSense fluorescent biomarker (N,N'-didansyl-L-cystine DDC) in vivo and subsequent fluorescence imaging of the intestinal mucosa. In the reported proof-of-concept studies, mice were treated with either taxol+cyclophosphamide or doxil. DDC was administered in vivo at various time points after drug administration, and tracer uptake by ileum tissue was subsequently evaluated by ex vivo fluorescent microscopy. Chemotherapy caused marked and selective uptake of DDC in ileal epithelial cells, in correlation with other hallmarks of apoptosis (i.e., DNA fragmentation and Annexin-V binding). Induction of DDC uptake occurred early after chemotherapy, and its temporal profile was parallel to that of the apoptotic process, as assessed histologically. DDC may therefore serve as a useful tool for detection of CIE. Future potential integration of this method with fluorescent endoscopic techniques, or development of radio-labeled derivatives of DDC for emission tomography, may advance early diagnosis and monitoring of this severe adverse effect of chemotherapy.

Cytidine-directed rapid synthesis of water-soluble and highly yellow fluorescent bimetallic AuAg nanoclusters.

PubMed

Zhang, Yuanyuan; Jiang, Hui; Ge, Wei; Li, Qiwei; Wang, Xuemei

2014-09-16

Fluorescent gold/silver nanoclusters templated by DNA or oligonucleotides have been widely reported since DNA or oligonucleotides could be designed to position a few metal ions at close proximity prior to their reduction, but nucleoside-templated synthesis is more challenging. In this work, a novel type of strategy taking cytidine (C) as template to rapid synthesis of fluorescent, water-soluble gold and silver nanoclusters (C-AuAg NCs) has been developed. The as-prepared C-AuAg NCs have been characterized by UV-vis absorption spectroscopy, fluorescence, transmission electron microscopy (TEM), energy dispersive X-ray spectroscopy (EDS), X-ray photoelectron spectroscopy (XPS), Fourier transform infrared spectroscopy (FT-IR), and inductively coupled plasma mass spectroscopy (ICP-MS). The characterizations demonstrate that C-AuAg NCs with a diameter of 1.50 ± 0.31 nm, a quantum yield ∼9%, and an average lifetime ∼6.07 μs possess prominent fluorescence properties, good dispersibility, and easy water solubility, indicating the promising application in bioanalysis and biomedical diagnosis. Furthermore, this strategy by rapid producing of highly fluorescent nanoclusters could be explored for the possible recognition of some disease-related changes in blood serum. This raises the possibility of their promising application in bioanalysis and biomedical diagnosis.

An agmatine-inducible system for the expression of recombinant proteins in Enterococcus faecalis.

PubMed

Linares, Daniel M; Perez, Marta; Ladero, Victor; Del Rio, Beatriz; Redruello, Begoña; Martin, M Cruz; Fernandez, María; Alvarez, Miguel A

2014-12-04

Scientific interest in Enterococcus faecalis has increased greatly over recent decades. Some strains are involved in food fermentation and offer health benefits, whereas others are vancomycin-resistant and cause infections that are difficult to treat. The limited availability of vectors able to express cloned genes efficiently in E. faecalis has hindered biotechnological studies on the bacterium's regulatory and pathogenicity-related genes. The agmatine deiminase (AGDI) pathway of E. faecalis, involved in the conversion of agmatine into putrescine, is driven by a response inducer gene aguR. This study describes that the exposure to the induction factor (agmatine) results in the transcription of genes under the control of the aguB promoter, including the aguBDAC operon. A novel E. faecalis expression vector, named pAGEnt, combining the aguR inducer gene and the aguB promoter followed by a cloning site and a stop codon was constructed. pAGEnt was designed for the overexpression and purification of a protein fused to a 10-amino-acid His-tag at the C-terminus. The use of GFP as a reporter of gene expression in E. faecalis revealed that under induction with 60 mM agmatine, fluorescence reached 40 arbitrary units compared to 0 in uninduced cells. pAGEnt vector can be used for the overexpression of recombinant proteins under the induction of agmatine in E. faecalis, with a close correlation between agmatine concentration and fluorescence when GFP was used as reporter.

75 FR 37975 - Energy Conservation Program for Consumer Products and Certain Commercial and Industrial Equipment...

Federal Register 2010, 2011, 2012, 2013, 2014

2010-07-01

... technologies including induction and fluorescent lamps. 75 FR 22031, 22033 (April 27, 2010). In comments... of magnetic ballasts of 50,000 hours. After the 2003 draft report, Allegheny noted that MV ballast... considered only magnetic ballasts because they are the most common ballast used in HID lighting systems. DOE...

Variable-frequency synchronous motor drives for electric vehicles

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

Chalmers, B.J.; Musaba, L.; Gosden, D.F.

1996-07-01

The performance capability envelope of a variable-frequency, permanent-magnet synchronous motor drive with field weakening is dependent upon the product of maximum current and direct-axis inductance. To obtain a performance characteristic suitable for a typical electric vehicle drive, in which short-term increase of current is applied, it is necessary to design an optimum value of direct-axis inductance. The paper presents an analysis of a hybrid motor design which uses a two-part rotor construction comprising a surface-magnet part and an axially laminated reluctance part. This arrangement combines the properties of all other types of synchronous motor and offers a greater choice ofmore » design variables. It is shown that the desired form of performance may be achieved when the high-inductance axis of the reluctance part is arranged to lead the magnet axis by 90{degree} (elec.).« less

Variable-frequency synchronous motor drives for electric vehicles

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

Chalmers, B.J.; Musaba, L.; Gosden, D.F.

1995-12-31

The performance capability envelope of a variable-frequency, permanent-magnet synchronous motor drive with field weakening is dependent upon the product of maximum current and direct-axis inductance. To obtain a performance characteristic suitable for a typical electric vehicle drive, in which short-term increase of current is applied, it is necessary to design an optimum value of direct-axis inductance. The paper presents an analysis of a hybrid motor design which uses a two-part rotor construction comprising a surface-magnet part and an axially-laminated reluctance part. This arrangement combines the properties of all other types of synchronous motor and offers a greater choice of designmore » variables. It is shown that the desired form of performance may be achieved when the high-inductance axis of the reluctance part is arranged to lead the magnet axis by 90{degree} (elec.).« less

Systems approach to excitation-energy and electron transfer reaction networks in photosystem II complex: model studies for chlorophyll a fluorescence induction kinetics.

PubMed

Matsuoka, Takeshi; Tanaka, Shigenori; Ebina, Kuniyoshi

2015-09-07

Photosystem II (PS II) is a protein complex which evolves oxygen and drives charge separation for photosynthesis employing electron and excitation-energy transfer processes over a wide timescale range from picoseconds to milliseconds. While the fluorescence emitted by the antenna pigments of this complex is known as an important indicator of the activity of photosynthesis, its interpretation was difficult because of the complexity of PS II. In this study, an extensive kinetic model which describes the complex and multi-timescale characteristics of PS II is analyzed through the use of the hierarchical coarse-graining method proposed in the authors׳ earlier work. In this coarse-grained analysis, the reaction center (RC) is described by two states, open and closed RCs, both of which consist of oxidized and neutral special pairs being in quasi-equilibrium states. Besides, the PS II model at millisecond scale with three-state RC, which was studied previously, could be derived by suitably adjusting the kinetic parameters of electron transfer between tyrosine and RC. Our novel coarse-grained model of PS II can appropriately explain the light-intensity dependent change of the characteristic patterns of fluorescence induction kinetics from O-J-I-P, which shows two inflection points, J and I, between initial point O and peak point P, to O-J-D-I-P, which shows a dip D between J and I inflection points. Copyright © 2015 Elsevier Ltd. All rights reserved.

Spectroscopy detection of green and red fluorescent proteins in genetically modified plants using a fiber optics system

NASA Astrophysics Data System (ADS)

Liew, Oi Wah; Asundi, Anand K.; Chen, Jun-Wei; Chew, Yiwen; Yu, Shangjuan; Yeo, Gare H.

2001-05-01

In this paper, fiber optic spectroscopy is developed to detect and quantify recombinant green (EGFP) and red (DsRED) fluorescent proteins in vitro and in vivo. The bacterial expression vectors carrying the coding regions of EGFP and DsRED were introduced into Escherichia coli host cells and fluorescent proteins were produced following induction with IPTG. Soluble EGFP and DsRED proteins were isolated from lysed bacterial cells and serially diluted for quantitative analysis by fiber optic spectroscopy. Fluorescence at the appropriate emission wavelengths could be detected up to 64X dilution for EGFP and 40X dilution for DsRED. To determine the capability of spectroscopy detection in vivo, transgenic potato hairy roots expressing EGFP and DsRED were regenerated. This was achieved by cloning the EGFP and DsRED genes into the plant binary vector, pTMV35S, to create the recombinant vectors pGLOWGreen and pGLOWRed. These latter binary vectors were introduced into Agrobacterium rhizogenes strain A4T. Infection of potato cells with transformed agrobacteria was used to insert the fluorescent protein genes into the potato genome. Genetically modified potato cells were then regenerated into hairy roots. A panel of transformed hairy roots expressing varying levels of fluorescent proteins was selected by fluorescence microscopy. We are now assessing the capability of spectroscopic detection system for in vivo quantification of green and red fluorescence levels in transformed roots.

Intracellular in situ labeling of TiO2 nanoparticles for fluorescence microscopy detection

PubMed Central

Brown, Koshonna; Thurn, Ted; Xin, Lun; Liu, William; Bazak, Remon; Chen, Si; Lai, Barry; Vogt, Stefan; Jacobsen, Chris; Paunesku, Tatjana; Woloschak, Gayle E.

2018-01-01

Titanium dioxide (TiO2) nanoparticles are produced for many different purposes, including development of therapeutic and diagnostic nanoparticles for cancer detection and treatment, drug delivery, induction of DNA double-strand breaks, and imaging of specific cells and subcellular structures. Currently, the use of optical microscopy, an imaging technique most accessible to biology and medical pathology, to detect TiO2 nanoparticles in cells and tissues ex vivo is limited with low detection limits, while more sensitive imaging methods (transmission electron microscopy, X-ray fluorescence microscopy, etc.) have low throughput and technical and operational complications. Herein, we describe two in situ post-treatment labeling approaches to stain TiO2 nanoparticles taken up by the cells. The first approach utilizes fluorescent biotin and fluorescent streptavidin to label the nanoparticles before and after cellular uptake; the second approach is based on the copper-catalyzed azide-alkyne cycloaddition, the so-called Click chemistry, for labeling and detection of azide-conjugated TiO2 nanoparticles with alkyne-conjugated fluorescent dyes such as Alexa Fluor 488. To confirm that optical fluorescence signals of these nanoparticles match the distribution of the Ti element, we used synchrotron X-ray fluorescence microscopy (XFM) at the Advanced Photon Source at Argonne National Laboratory. Titanium-specific XFM showed excellent overlap with the location of optical fluorescence detected by confocal microscopy. Therefore, future experiments with TiO2 nanoparticles may safely rely on confocal microscopy after in situ nanoparticle labeling using approaches described here. PMID:29541425

Intracellular in situ labeling of TiO2 nanoparticles for fluorescence microscopy detection.

PubMed

Brown, Koshonna; Thurn, Ted; Xin, Lun; Liu, William; Bazak, Remon; Chen, Si; Lai, Barry; Vogt, Stefan; Jacobsen, Chris; Paunesku, Tatjana; Woloschak, Gayle E

2018-01-01

Titanium dioxide (TiO 2 ) nanoparticles are produced for many different purposes, including development of therapeutic and diagnostic nanoparticles for cancer detection and treatment, drug delivery, induction of DNA double-strand breaks, and imaging of specific cells and subcellular structures. Currently, the use of optical microscopy, an imaging technique most accessible to biology and medical pathology, to detect TiO 2 nanoparticles in cells and tissues ex vivo is limited with low detection limits, while more sensitive imaging methods (transmission electron microscopy, X-ray fluorescence microscopy, etc.) have low throughput and technical and operational complications. Herein, we describe two in situ post-treatment labeling approaches to stain TiO 2 nanoparticles taken up by the cells. The first approach utilizes fluorescent biotin and fluorescent streptavidin to label the nanoparticles before and after cellular uptake; the second approach is based on the copper-catalyzed azide-alkyne cycloaddition, the so-called Click chemistry, for labeling and detection of azide-conjugated TiO 2 nanoparticles with alkyne-conjugated fluorescent dyes such as Alexa Fluor 488. To confirm that optical fluorescence signals of these nanoparticles match the distribution of the Ti element, we used synchrotron X-ray fluorescence microscopy (XFM) at the Advanced Photon Source at Argonne National Laboratory. Titanium-specific XFM showed excellent overlap with the location of optical fluorescence detected by confocal microscopy. Therefore, future experiments with TiO 2 nanoparticles may safely rely on confocal microscopy after in situ nanoparticle labeling using approaches described here.

Pinocytosis and locomotion of amoebae. XV. Visualization of Ca++-dynamics by chlorotetracycline (CTC) fluorescence during induced pinocytosis in living Amoeba proteus.

PubMed

Gawlitta, W; Stockem, W; Wehland, J; Weber, K

1980-01-01

The dynamics of Ca++ during induced pinocytosis were studied in Amoeba proteus using chlorotetracycline (CTC). The fluorescence of the Ca++ - CTC-complex was monitored by an image intensification system, which has certain advantages over standard equipment: (1) Living cells are not subjected to the damaging influence of intensive microscopic illumination, (2) fluorescent probes are not bleached during observation, and (3) the rapid dynamics of the Ca++ -fluxes can be recorded using short exposure times. The results demonstrate the existence of Ca++ bound to intracellular and extracellular sites of the cell membrane complex in normal locomoting and pinocytotic Amoeba proteus. The application of cations inducing pinocytosis causes a rapid decrease in the external CTC-fluorescence probably due to a release of Ca++ from the mucous layer. The degree of fluorescence intensity is correlated with the capacity of pinocytotic channel formation, i.e., the fluorescence decreases as the number of channels increases. During the phase of vesiculation a distinct fluorescence mainly restricted to the basal region of the channels is observed. Intracellular Ca++ was detected in close vicinity to the plasma membrane after both microinjection and external application of CTC. The internal CTC-fluorescence is slightly decreased during the induction phase of pinocytosis. The observations are in good agreement with previous results on the localization of Ca++ -binding sites at the plasma membrane of Amoeba proteus and demonstrate the important role of Ca++ -fluxes for the process of pinocytosis.

Miniaturized fiber-coupled confocal fluorescence microscope with an electrowetting variable focus lens using no moving parts

PubMed Central

Ozbay, Baris N.; Losacco, Justin T.; Cormack, Robert; Weir, Richard; Bright, Victor M.; Gopinath, Juliet T.; Restrepo, Diego; Gibson, Emily A.

2015-01-01

We report a miniature, lightweight fiber-coupled confocal fluorescence microscope that incorporates an electrowetting variable focus lens to provide axial scanning for full three-dimensional (3D) imaging. Lateral scanning is accomplished by coupling our device to a laser-scanning confocal microscope through a coherent imaging fiber-bundle. The optical components of the device are combined in a custom 3D-printed adapter with an assembled weight of <2 g that can be mounted onto the head of a mouse. Confocal sectioning provides an axial resolution of ~12 µm and an axial scan range of ~80 µm. The lateral field-of-view is 300 µm, and the lateral resolution is 1.8 µm. We determined these parameters by imaging fixed sections of mouse neuronal tissue labeled with green fluorescent protein (GFP) and fluorescent bead samples in agarose gel. To demonstrate viability for imaging intact tissue, we resolved multiple optical sections of ex vivo mouse olfactory nerve fibers expressing yellow fluorescent protein (YFP). PMID:26030555

Variable speed induction motor operation from a 20-kHz power bus

NASA Technical Reports Server (NTRS)

Hansen, Irving G.

1989-01-01

Induction motors are recognized for their simple rugged construction. To date, however, their application to variable speed or servo drives was hampered by limitations on their control. Induction motor drives tend to be complex and to display troublesome low speed characteristics due in part to nonsinusoidal driving voltages. A technique was developed which involves direct synthesis of sinusoidal driving voltages from a high frequency power bus and independent control of frequency and voltages. Separation of frequency and voltage allows independent control of rotor and stator flux, full four quadrant operation, and instantaneous torque control. Recent test results, current status of the technology, and proposed aerospace applications will be discussed.

Variable speed induction motor operation from a 20-kHz power bus

NASA Technical Reports Server (NTRS)

Hansen, Irving G.

1989-01-01

Induction motors are recognized for their simple rugged construction to date, however, their application to variable speed or servo drives has been hampered by limitations on their control. Induction motor drives tend to be complex and to display troublesome low speed characteristics due in part to nonsinusoidal driving voltages. A technique was developed which involves direct synthesis of sinusoidal driving voltages from a high frequency power bus and independent control of frequency and voltages. Separation offrequency and voltage allows independent control of rotor and stator flux, full four-quadrant operation, and instantaneous torque control. Recent test results, current status of the technology, and proposed aerospace applications will be discussed.

Modular fluorescence complementation sensors for live cell detection of epigenetic signals at endogenous genomic sites.

PubMed

Lungu, Cristiana; Pinter, Sabine; Broche, Julian; Rathert, Philipp; Jeltsch, Albert

2017-09-21

Investigation of the fundamental role of epigenetic processes requires methods for the locus-specific detection of epigenetic modifications in living cells. Here, we address this urgent demand by developing four modular fluorescence complementation-based epigenetic biosensors for live-cell microscopy applications. These tools combine engineered DNA-binding proteins with domains recognizing defined epigenetic marks, both fused to non-fluorescent fragments of a fluorescent protein. The presence of the epigenetic mark at the target DNA sequence leads to the reconstitution of a functional fluorophore. With this approach, we could for the first time directly detect DNA methylation and histone 3 lysine 9 trimethylation at endogenous genomic sites in live cells and follow dynamic changes in these marks upon drug treatment, induction of epigenetic enzymes and during the cell cycle. We anticipate that this versatile technology will improve our understanding of how specific epigenetic signatures are set, erased and maintained during embryonic development or disease onset.Tools for imaging epigenetic modifications can shed light on the regulation of epigenetic processes. Here, the authors present a fluorescence complementation approach for detection of DNA and histone methylation at endogenous genomic sites allowing following of dynamic changes of these marks by live-cell microscopy.

Induction of neonatal tolerance to GFP-labeled karyocytes in C57/B6 mice.

PubMed

Dovhyi, Roman; Pishel, Iryna; Butenko, Gennadij; Skivka, Larysa

2017-08-01

Green fluorescent protein is widely used in biological studies including parabiosis models for visualization of cellular structures and cells. However, the growing number of the data is available regarding immunogenicity of this protein, which can interfere with its use in in vivo experiments. In this study, we attempted to induce neonatal immunological tolerance to GFP-labeled karyocytes by intraperitoneal injections of B6.GFP mouse splenocytes to newborn C57/B6 mice. GFP + skin graft integrity was evaluated under UV light at 6weeks after skin grafting. GFP + skin transplants survived up to 6weeks after grafting in all animals that undergone neonatal tolerance induction, whereas all skin grafts were rejected in control naïve mice within first two weeks. Thus, current protocol is suitable for induction of immune tolerance against GFP-labeled karyocytes. Copyright © 2017 Elsevier B.V. All rights reserved.

Evaluation of hyperaccumulator plant species grown in metalliferous sites in Albania

NASA Astrophysics Data System (ADS)

Babani, F.; Civici, N.; Mullaj, A.; Kongjika, E.; Ylli, A.

2007-04-01

Heavy metal contamination of soils causes serious problems to our society. A small number of interesting plant species have been identified that can grow in soils containing high levels of heavy metals, and can also accumulate these metals to high concentrations in the shoot. The heavy metal contents in root, shoot, leaves and flowers of spontaneous plants grown in metalliferous sites in Albania together with the elemental composition of the native soils were determined by X-ray fluorescence spectrometry. Efficiency of photosynthetic apparatus of analyzed ecotypes was evaluated via chlorophyll fluorescence imaging during induction kinetics. Response of plant root system to the presence of metals, the available pools of metals to plants, effect of plant biomass to phytoextraction, photosynthetic pigment metabolism and chlorophyll fluorescence signature of leaves allowed to characterize hyperaccumulator properties and to detect the variation between selected ecotypes to heavy metal accumulation.

Portable plant chlorophyll fluorimeter based on blue LED rapid induced technology

NASA Astrophysics Data System (ADS)

Zheng, Yibo; Mi, Ting; Zhang, Lei; Zhao, Jun

2018-01-01

Fluorimeter is an effective device for detecting chlorophyll a content in plants. In order to realize real-time nondestructive detection of plant blades, a camera based fluorescence instrument based on two color mirrors has been developed. The blue light LED is used as the excitation light source, and the lens is used for shaping and focusing the excitation light to ensure the excitation intensity and uniform illumination of the light source. The device uses a 45 degree two color mirror to separate the chlorophyll a excited light path and the fluorescence receiving light path. Finally, the fluorescent signal is collected by the silicon photocell, and the signal is processed by the circuit to transmit the digital information to the display. Through the analysis of the experimental data, the device has the advantages of small size, easy to carry, fast induction, etc., and can be widely applied in outdoor teaching and field investigation.

Heterogeneous accumulation of fluorescent bile acids in primary rat hepatocytes does not correlate with their homogenous expression of ntcp.

PubMed

Murray, John W; Thosani, Amar J; Wang, Pijun; Wolkoff, Allan W

2011-07-01

Sodium taurocholate-cotransporting polypeptide (ntcp) is considered to be a major determinant of bile acid uptake into hepatocytes. However, the regulation of ntcp and the degree that it participates in the accumulation of specific substrates are not well understood. We utilized fluorescent bile acid derivatives and direct quantitation of fluorescent microscopy images to examine the regulation of ntcp and its role in the cell-to-cell variability of fluorescent bile acid accumulation. Primary-cultured rat hepatocytes rapidly accumulated the fluorescent bile acids, chenodeoxycholylglycylamidofluorescein (CDCGamF), 7-β- nitrobenzoxadiazole 3-α hydroxy 5-β cholan-24-oic acid (NBD-CA), and cholyl-glycylamido-fluorescein (CGamF). However, in stably transfected HeLa cells, ntcp preferred CDCGamF, whereas the organic anion transporter, organic anion transporting polypeptide 1 (oatp1a1), preferred NBD-CA, and neither ntcp nor oatp1a1 showed strong accumulation of CGamF by these methods. Ntcp-mediated transport of CDCGamF was inhibited by taurocholate, cyclosporin, actin depolymerization, and an inhibitor of atypical PKC-ζ. The latter two agents altered the cellular distribution of ntcp as visualized in ntcp-green fluorescent protein-transfected cells. Although fluorescent bile acid accumulation was reproducible by the imaging assays, individual cells showed variable accumulation that was not attributable to changes in membrane permeability or cell viability. In HeLa cells, this was accounted for by variable levels of ntcp, whereas, in hepatocytes, ntcp expression was uniform, and low accumulation was seen in a large portion of cells despite the presence of ntcp. These studies indicate that single-cell imaging can provide insight into previously unrecognized details of anion transport in the complex environment of polarized hepatocytes.

Heterogeneous accumulation of fluorescent bile acids in primary rat hepatocytes does not correlate with their homogenous expression of ntcp

PubMed Central

Thosani, Amar J.; Wang, Pijun; Wolkoff, Allan W.

2011-01-01

Sodium taurocholate-cotransporting polypeptide (ntcp) is considered to be a major determinant of bile acid uptake into hepatocytes. However, the regulation of ntcp and the degree that it participates in the accumulation of specific substrates are not well understood. We utilized fluorescent bile acid derivatives and direct quantitation of fluorescent microscopy images to examine the regulation of ntcp and its role in the cell-to-cell variability of fluorescent bile acid accumulation. Primary-cultured rat hepatocytes rapidly accumulated the fluorescent bile acids, chenodeoxycholylglycylamidofluorescein (CDCGamF), 7-β- nitrobenzoxadiazole 3-α hydroxy 5-β cholan-24-oic acid (NBD-CA), and cholyl-glycylamido-fluorescein (CGamF). However, in stably transfected HeLa cells, ntcp preferred CDCGamF, whereas the organic anion transporter, organic anion transporting polypeptide 1 (oatp1a1), preferred NBD-CA, and neither ntcp nor oatp1a1 showed strong accumulation of CGamF by these methods. Ntcp-mediated transport of CDCGamF was inhibited by taurocholate, cyclosporin, actin depolymerization, and an inhibitor of atypical PKC-ζ. The latter two agents altered the cellular distribution of ntcp as visualized in ntcp-green fluorescent protein-transfected cells. Although fluorescent bile acid accumulation was reproducible by the imaging assays, individual cells showed variable accumulation that was not attributable to changes in membrane permeability or cell viability. In HeLa cells, this was accounted for by variable levels of ntcp, whereas, in hepatocytes, ntcp expression was uniform, and low accumulation was seen in a large portion of cells despite the presence of ntcp. These studies indicate that single-cell imaging can provide insight into previously unrecognized details of anion transport in the complex environment of polarized hepatocytes. PMID:21474652

Photorhabdus luminescens genes induced upon insect infection

PubMed Central

Münch, Anna; Stingl, Lavinia; Jung, Kirsten; Heermann, Ralf

2008-01-01

Background Photorhabdus luminescens is a Gram-negative luminescent enterobacterium and a symbiote to soil nematodes belonging to the species Heterorhabditis bacteriophora. P.luminescens is simultaneously highly pathogenic to insects. This bacterium exhibits a complex life cycle, including one symbiotic stage characterized by colonization of the upper nematode gut, and a pathogenic stage, characterized by release from the nematode into the hemocoel of insect larvae, resulting in rapid insect death caused by bacterial toxins. P. luminescens appears to sense and adapt to the novel host environment upon changing hosts, which facilitates the production of factors involved in survival within the host, host-killing, and -exploitation. Results A differential fluorescence induction (DFI) approach was applied to identify genes that are up-regulated in the bacterium after infection of the insect host Galleria mellonella. For this purpose, a P. luminescens promoter-trap library utilizing the mCherry fluorophore as a reporter was constructed, and approximately 13,000 clones were screened for fluorescence induction in the presence of a G. mellonella larvae homogenate. Since P. luminescens has a variety of regulators that potentially sense chemical molecules, like hormones, the screen for up-regulated genes or operons was performed in vitro, excluding physicochemical signals like oxygen, temperature or osmolarity as variables. Clones (18) were obtained exhibiting at least 2.5-fold induced fluorescence and regarded as specific responders to insect homogenate. In combination with a bioinformatics approach, sequence motifs were identified in these DNA-fragments that are similar to 29 different promoters within the P. luminescens genome. By cloning each of the predicted promoters upstream of the reporter gene, induction was verified for 27 promoters in vitro, and for 24 promoters in viable G. mellonella larvae. Among the validated promoters are some known to regulate the expression of toxin genes, including tccC1 (encoding an insecticidal toxin complex), and others encoding putative toxins. A comparably high number of metabolic genes or operons were observed to be induced upon infection; among these were eutABC, hutUH, and agaZSVCD, which encode proteins involved in ethanolamine, histidine and tagatose degradation, respectively. The results reflect rearrangements in metabolism and the use of other metabolites available from the insect. Furthermore, enhanced activity of promoters controlling the expression of genes encoding enzymes linked to antibiotic production and/or resistance was observed. Antibiotic production and resistance may influence competition with other bacteria, and thus might be important for a successful infection. Lastly, several genes of unknown function were identified that may represent novel pathogenicity factors. Conclusion We show that a DFI screen is useful for identifying genes or operons induced by chemical stimuli, such as diluted insect homogenate. A bioinformatics comparison of motifs similar to known promoters is a powerful tool for identifying regulated genes or operons. We conclude that signals for the regulation of those genes or operons induced in P. luminescens upon insect infection may represent a wide variety of compounds that make up the insect host. Our results provide insight into the complex response to the host that occurs in a bacterial pathogen, particularly reflecting the potential for metabolic shifts and other specific changes associated with virulence. PMID:18489737

Induction signals from Callisto's ionosphere and their implications on a possible subsurface ocean

NASA Astrophysics Data System (ADS)

Hartkorn, Oliver; Saur, Joachim

2017-11-01

We investigate whether induction within Callisto's electrically conductive ionosphere can explain observed magnetic fields which have previously been interpreted as evidence of induction in a saline, electrically conductive subsurface ocean. Callisto's ionosphere is subject to the flow of time-periodic magnetized plasma of Jupiter's magnetosphere, which induces electric fields and electric currents in Callisto's electrically conductive ionosphere. We develop a simple analytic model for a first quantitative understanding of the effects of induction in Callisto's ionosphere caused by the interaction with a time-variable magnetic field environment. With this model, we also investigate how the associated ionospheric currents close in the ambient magnetospheric plasma. Based on our model, we find that the anisotropic nature of Callisto's ionospheric conductivity generates an enhancement effect on ionospheric loop currents which are driven by the time-variable magnetic field. This effect is similar to the Cowling channel effect known from Earth's ionosphere. Subsequently, we numerically calculate the expected induced magnetic fields due to Jupiter's time-variable magnetic field in an anisotropic conductive ionosphere and compare our results with the Galileo C-3 and C-9 flybys. We find that induction within Callisto's ionosphere is responsible for a significant part of the observed magnetic fields. Ionospheric induction creates induced magnetic fields to some extent similar as expected from a subsurface water ocean. Depending on currently unknown properties such as Callisto's nightside ionosphere, the existence of layers of "dirty ice" and the details of the plasma interaction, a water ocean might be located much deeper than previously thought or might not exist at all.

Effects of gold nanoparticles on the photophysical and photosynthetic parameters of leaves and chloroplasts.

PubMed

Torres, Rocio; Diz, Virginia E; Lagorio, M Gabriela

2018-04-18

Effects of gold nanoparticles (average diameter: 10-14 nm) on leaves and chloroplasts have been studied. Gold nanoparticles (AuNPs) quenched significantly chlorophyll fluorescence when introduced both in intact leaves and isolated chloroplasts. Additionally, the fluorescence spectra corrected for light re-absorption processes showed a net decrease in the fluorescence ratio calculated as the quotient between the maximum fluorescence at 680 and 735 nm. This fact gave evidence for a reduction in the fluorescence emission of the PSII relative to that of the PSI. Strikingly, the photosynthetic parameters derived from the analysis of the slow phase of Kautsky's kinetics, the rate of oxygen evolution and the rate of photo-reduction of 2,6-dichlorophenolindophenol were increased in the presence of AuNPs indicating an apparent greater photosynthetic capacity. The observed results were consistent with an electron transfer process from the excited PSII, which was thermodynamically possible, and which competed with both the electron transport process that initiated photosynthesis and the deactivation of the excited PSII by fluorescence emission. Additionally, it is here explained, in terms of a completely rational kinetic scheme and their corresponding algebraic expressions, why the photosynthetic parameters and the variable and non-variable fluorescence of chlorophyll are modified in a photosynthetic tissue containing gold nanoparticles.

Synthesis of yellow and red fluorescent 1,3a,6a-triazapentalenes and the theoretical investigation of their optical properties† †Electronic supplementary information (ESI) available: the experimental details for the synthesis of the triazapentalenes and the fluorescent cell staining, the absorption and fluorescence spectra, and the 1H and 13C NMR spectra. Also given are the molecular orbitals, the natural charges, the dipole moments, and the Cartesian coordinates of the triazapentalenes (1a, 1b, 1g, 1e, and 1f). See DOI: 10.1039/c4sc02780a Click here for additional data file.

PubMed Central

Osawa, Ayumi; Mera, Akane; Tano, Fumi; Chuman, Yoshiro; Sakuda, Eri; Taketsugu, Tetsuya; Sakaguchi, Kazuyasu; Kitamura, Noboru

2015-01-01

To expand the originally developed fluorescent 1,3a,6a-triazapentalenes as fluorescent labelling reagents, the fluorescence wavelength of the 1,3a,6a-triazapentalenes was extended to the red color region. Based on the noteworthy correlation of the fluorescence wavelength with the inductive effect of the 2-substituent, electron-deficient 2-(2-cyano-4-methoxycarbonylphenyl)-1,3a,6a-triazapentalene and 2-(2,6-dicyano-4-methoxycarbonylphenyl)-1,3a,6a-triazapentalene were synthesized. The former exhibited yellow fluorescence and the latter exhibited red fluorescence, and both compounds exhibited large Stokes shifts, and the 1,3a,6a-triazapentalene system enabled the same fluorescent chromophore to cover the entire region of visible wavelengths. The potential applications of the 1,3a,6a-triazapentalenes as fluorescent probes in the fields of the life sciences were investigated, and the 1,3a,6a-triazapentalene system was clearly proven to be useful as a fluorescent reagent for live cell imaging. Quantum chemical calculations were performed to investigate the optical properties of the 1,3a,6a-triazapentalenes. These calculations revealed that the excitation involves a significant charge-transfer from the 1,3a,6a-triazapentalene skeleton to the 2-substituent. The calculated absorption and fluorescence wavelengths showed a good correlation with the experimental ones, and thus the system could enable the theoretical design of substituents with the desired optical properties. PMID:29560196

Effects of acepromazine and trazodone on anesthetic induction dose of propofol and cardiovascular variables in dogs undergoing general anesthesia for orthopedic surgery.

PubMed

Murphy, Lindsey A; Barletta, Michele; Graham, Lynelle F; Reichl, Lorna J; Duxbury, Margaret M; Quandt, Jane E

2017-02-15

OBJECTIVE To compare the doses of propofol required to induce general anesthesia in dogs premedicated with acepromazine maleate or trazodone hydrochloride and compare the effects of these premedicants on cardiovascular variables in dogs anesthetized for orthopedic surgery. DESIGN Prospective, randomized study. ANIMALS 30 systemically healthy client-owned dogs. PROCEDURES 15 dogs received acepromazine (0.01 to 0.03 mg/kg [0.005 to 0.014 mg/lb], IM) 30 minutes before anesthetic induction and 15 received trazodone (5 mg/kg [2.27 mg/lb] for patients > 10 kg or 7 mg/kg [3.18 mg/lb] for patients ≤ 10 kg, PO) 2 hours before induction. Both groups received morphine sulfate (1 mg/kg [0.45 mg/lb], IM) 30 minutes before induction. Anesthesia was induced with propofol (4 to 6 mg/kg [1.82 to 2.73 mg/lb], IV, to effect) and maintained with isoflurane or sevoflurane in oxygen. Bupivacaine (0.5 mg/kg [0.227 mg/lb]) and morphine (0.1 mg/kg [0.045 mg/lb]) were administered epidurally. Dogs underwent tibial plateau leveling osteotomy (n = 22) or tibial tuberosity advancement (8) and were monitored throughout anesthesia. Propofol induction doses and cardiovascular variables (heart rate and systemic, mean, and diastolic arterial blood pressures) were compared between groups. RESULTS The mean dose of propofol required for anesthetic induction and all cardiovascular variables evaluated did not differ between groups. Intraoperative hypotension developed in 6 and 5 dogs of the acepromazine and trazodone groups, respectively; bradycardia requiring intervention developed in 3 dogs/group. One dog that received trazodone had priapism 24 hours later and was treated successfully. No other adverse effects were reported. CONCLUSIONS AND CLINICAL RELEVANCE At the described dosages, cardiovascular effects of trazodone were similar to those of acepromazine in healthy dogs undergoing anesthesia for orthopedic surgery.

Drill core major, trace and rare earth element anlayses from wells RN-17B and RN-30, Reykjanes, Iceland

DOE Data Explorer

Andrew Fowler

2015-04-01

Analytical results for X-ray fluorescence (XRF) and inductively coupled plasma mass spectrometry (ICP-MS) measurement of major, trace and rare earth elements in drill core from geothermal wells in Reykjanes, Iceland. Total Fe was analyzed as FeO, therefore is not included under the Fe2O3 column.

Drill cutting and core major, trace and rare earth element anlayses from wells RN-17B and RN-30, Reykjanes, Iceland

DOE Data Explorer

Andrew Fowler

2015-05-01

Analytical results for x-ray fluorescence (XRF) and Inductively Couple Plasma Mass Spectrometry (ICP-MS) measurement of major, trace and rare earth elements in drill cuttings from geothermal wells in Reykjanes, Iceland. Total Fe was analyzed as FeO, therefore is not included under the Fe2O3 column.

Uptake of bright fluorophore core-silica shell nanoparticles by biological systems

PubMed Central

Zane, Andrew; McCracken, Christie; Knight, Deborah A; Young, Tanya; Lutton, Anthony D; Olesik, John W; Waldman, W James; Dutta, Prabir K

2015-01-01

Nanoparticles are used in a variety of consumer applications. Silica nanoparticles in particular are common, including as a component of foods. There are concerns that ingested nano-silica particles can cross the intestinal epithelium, enter the circulation, and accumulate in tissues and organs. Thus, tracking these particles is of interest, and fluorescence spectroscopic methods are well-suited for this purpose. However, nanosilica is not fluorescent. In this article, we focus on core-silica shell nanoparticles, using fluorescent Rhodamine 6G, Rhodamine 800, or CdSe/CdS/ZnS quantum dots as the core. These stable fluorophore/silica nanoparticles had surface characteristics similar to those of commercial silica particles. Thus, they were used as model particles to examine internalization by cultured cells, including an epithelial cell line relevant to the gastrointestinal tract. Finally, these particles were administered to mice by gavage, and their presence in various organs, including stomach, small intestine, cecum, colon, kidney, lung, brain, and spleen, was examined. By combining confocal fluorescence microscopy with inductively coupled plasma mass spectrometry, the presence of nanoparticles, rather than their dissolved form, was established in liver tissues. PMID:25759579

Nepenthesin protease activity indicates digestive fluid dynamics in carnivorous nepenthes plants.

PubMed

Buch, Franziska; Kaman, Wendy E; Bikker, Floris J; Yilamujiang, Ayufu; Mithöfer, Axel

2015-01-01

Carnivorous plants use different morphological features to attract, trap and digest prey, mainly insects. Plants from the genus Nepenthes possess specialized leaves called pitchers that function as pitfall-traps. These pitchers are filled with a digestive fluid that is generated by the plants themselves. In order to digest caught prey in their pitchers, Nepenthes plants produce various hydrolytic enzymes including aspartic proteases, nepenthesins (Nep). Knowledge about the generation and induction of these proteases is limited. Here, by employing a FRET (fluorescent resonance energy transfer)-based technique that uses a synthetic fluorescent substrate an easy and rapid detection of protease activities in the digestive fluids of various Nepenthes species was feasible. Biochemical studies and the heterologously expressed Nep II from Nepenthes mirabilis proved that the proteolytic activity relied on aspartic proteases, however an acid-mediated auto-activation mechanism was necessary. Employing the FRET-based approach, the induction and dynamics of nepenthesin in the digestive pitcher fluid of various Nepenthes plants could be studied directly with insect (Drosophila melanogaster) prey or plant material. Moreover, we observed that proteolytic activity was induced by the phytohormone jasmonic acid but not by salicylic acid suggesting that jasmonate-dependent signaling pathways are involved in plant carnivory.

Nepenthesin Protease Activity Indicates Digestive Fluid Dynamics in Carnivorous Nepenthes Plants

PubMed Central

Buch, Franziska; Kaman, Wendy E.; Bikker, Floris J.; Yilamujiang, Ayufu; Mithöfer, Axel

2015-01-01

Carnivorous plants use different morphological features to attract, trap and digest prey, mainly insects. Plants from the genus Nepenthes possess specialized leaves called pitchers that function as pitfall-traps. These pitchers are filled with a digestive fluid that is generated by the plants themselves. In order to digest caught prey in their pitchers, Nepenthes plants produce various hydrolytic enzymes including aspartic proteases, nepenthesins (Nep). Knowledge about the generation and induction of these proteases is limited. Here, by employing a FRET (fluorescent resonance energy transfer)-based technique that uses a synthetic fluorescent substrate an easy and rapid detection of protease activities in the digestive fluids of various Nepenthes species was feasible. Biochemical studies and the heterologously expressed Nep II from Nepenthes mirabilis proved that the proteolytic activity relied on aspartic proteases, however an acid-mediated auto-activation mechanism was necessary. Employing the FRET-based approach, the induction and dynamics of nepenthesin in the digestive pitcher fluid of various Nepenthes plants could be studied directly with insect (Drosophila melanogaster) prey or plant material. Moreover, we observed that proteolytic activity was induced by the phytohormone jasmonic acid but not by salicylic acid suggesting that jasmonate-dependent signaling pathways are involved in plant carnivory. PMID:25750992

Tungsten devices in analytical atomic spectrometry

NASA Astrophysics Data System (ADS)

Hou, Xiandeng; Jones, Bradley T.

2002-04-01

Tungsten devices have been employed in analytical atomic spectrometry for approximately 30 years. Most of these atomizers can be electrically heated up to 3000 °C at very high heating rates, with a simple power supply. Usually, a tungsten device is employed in one of two modes: as an electrothermal atomizer with which the sample vapor is probed directly, or as an electrothermal vaporizer, which produces a sample aerosol that is then carried to a separate atomizer for analysis. Tungsten devices may take various physical shapes: tubes, cups, boats, ribbons, wires, filaments, coils and loops. Most of these orientations have been applied to many analytical techniques, such as atomic absorption spectrometry, atomic emission spectrometry, atomic fluorescence spectrometry, laser excited atomic fluorescence spectrometry, metastable transfer emission spectroscopy, inductively coupled plasma optical emission spectrometry, inductively coupled plasma mass spectrometry and microwave plasma atomic spectrometry. The analytical figures of merit and the practical applications reported for these techniques are reviewed. Atomization mechanisms reported for tungsten atomizers are also briefly summarized. In addition, less common applications of tungsten devices are discussed, including analyte preconcentration by adsorption or electrodeposition and electrothermal separation of analytes prior to analysis. Tungsten atomization devices continue to provide simple, versatile alternatives for analytical atomic spectrometry.

Input Power Characteristics of the Thyristor Variable Voltage Power Conditioner

DOT National Transportation Integrated Search

1973-11-01

A laboratory study was made of transformer and thyristor voltage control for speed control of a rotary induction motor. The test program consisted of two parts; the first dealing with measurements of the induction motor characteristics and the second...

Survey of the occurrence of desiccation-induced quenching of basal fluorescence in 28 species of green microalgae.

PubMed

Wieners, Paul Christian; Mudimu, Opayi; Bilger, Wolfgang

2018-05-30

Desiccation-induced chlorophyll fluorescence quenching seems to be an indispensable part of desiccation resistance in the surveyed 28 green microalgal species. Lichens are desiccation tolerant meta-organisms. In the desiccated state photosynthesis is inhibited rendering the photobionts potentially sensitive to photoinhibition. As a photoprotective mechanism, strong non-radiative dissipation of absorbed light leading to quenching of chlorophyll fluorescence has been proposed. Desiccation-induced quenching affects not only variable fluorescence, but also the so-called basal fluorescence, F 0 . This phenomenon is well-known for intact lichens and some free living aero-terrestrial algae, but it was often absent in isolated lichen algae. Therefore, a thorough screening for the appearance of desiccation-induced quenching was undertaken with 13 different aero-terrestrial microalgal species and lichen photobionts. They were compared with 15 aquatic green microalgal species, among them also three marine species. We asked the following questions: Do isolated lichen algae show desiccation-induced quenching? Are aero-terrestrial algae different in this respect to aquatic algae and is the potential for desiccation-induced quenching coupled to desiccation tolerance? How variable is desiccation-induced quenching among species? Most of the aero-terrestrial algae, including all lichen photobionts, showed desiccation-induced quenching, although highly variable in extent, whereas most of the aquatic algae did not. All algae displaying quenching were also desiccation tolerant, whereas all algae unable to perform desiccation-induced quenching were desiccation intolerant. Desiccation-induced fluorescence quenching seems to be an indispensable part of desiccation resistance in the investigated species.

Intracellular in situ labeling of TiO 2 nanoparticles for fluorescence microscopy detection

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

Brown, Koshonna; Thurn, Ted; Xin, Lun

Titanium dioxide (TiO 2) nanoparticles are produced for many different purposes, including development of therapeutic and diagnostic nanoparticles for cancer detection and treatment, drug delivery, induction of DNA double-strand breaks, and imaging of specific cells and subcellular structures. Currently, the use of optical microscopy, an imaging technique most accessible to biology and medical pathology, to detect TiO 2 nanoparticles in cells and tissues ex vivo is limited with low detection limits, while more sensitive imaging methods (transmission electron microscopy, X-ray fluorescence microscopy, etc.) have low throughput and technical and operational complications. In this paper, we describe two in situ posttreatmentmore » labeling approaches to stain TiO 2 nanoparticles taken up by the cells. The first approach utilizes fluorescent biotin and fluorescent streptavidin to label the nanoparticles before and after cellular uptake; the second approach is based on the copper-catalyzed azide-alkyne cycloaddition, the so-called Click chemistry, for labeling and detection of azide-conjugated TiO 2 nanoparticles with alkyneconjugated fluorescent dyes such as Alexa Fluor 488. To confirm that optical fluorescence signals of these nanoparticles match the distribution of the Ti element, we used synchrotron X-ray fluorescence microscopy (XFM) at the Advanced Photon Source at Argonne National Laboratory. Titanium-specific XFM showed excellent overlap with the location of optical fluorescence detected by confocal microscopy. Finally and therefore, future experiments with TiO 2 nanoparticles may safely rely on confocal microscopy after in situ nanoparticle labeling using approaches described here.« less

Intracellular in situ labeling of TiO 2 nanoparticles for fluorescence microscopy detection

DOE PAGES

Brown, Koshonna; Thurn, Ted; Xin, Lun; ...

2017-07-19

Titanium dioxide (TiO 2) nanoparticles are produced for many different purposes, including development of therapeutic and diagnostic nanoparticles for cancer detection and treatment, drug delivery, induction of DNA double-strand breaks, and imaging of specific cells and subcellular structures. Currently, the use of optical microscopy, an imaging technique most accessible to biology and medical pathology, to detect TiO 2 nanoparticles in cells and tissues ex vivo is limited with low detection limits, while more sensitive imaging methods (transmission electron microscopy, X-ray fluorescence microscopy, etc.) have low throughput and technical and operational complications. In this paper, we describe two in situ posttreatmentmore » labeling approaches to stain TiO 2 nanoparticles taken up by the cells. The first approach utilizes fluorescent biotin and fluorescent streptavidin to label the nanoparticles before and after cellular uptake; the second approach is based on the copper-catalyzed azide-alkyne cycloaddition, the so-called Click chemistry, for labeling and detection of azide-conjugated TiO 2 nanoparticles with alkyneconjugated fluorescent dyes such as Alexa Fluor 488. To confirm that optical fluorescence signals of these nanoparticles match the distribution of the Ti element, we used synchrotron X-ray fluorescence microscopy (XFM) at the Advanced Photon Source at Argonne National Laboratory. Titanium-specific XFM showed excellent overlap with the location of optical fluorescence detected by confocal microscopy. Finally and therefore, future experiments with TiO 2 nanoparticles may safely rely on confocal microscopy after in situ nanoparticle labeling using approaches described here.« less

Variable selection based on clustering analysis for improvement of polyphenols prediction in green tea using synchronous fluorescence spectra

NASA Astrophysics Data System (ADS)

Shan, Jiajia; Wang, Xue; Zhou, Hao; Han, Shuqing; Riza, Dimas Firmanda Al; Kondo, Naoshi

2018-04-01

Synchronous fluorescence spectra, combined with multivariate analysis were used to predict flavonoids content in green tea rapidly and nondestructively. This paper presented a new and efficient spectral intervals selection method called clustering based partial least square (CL-PLS), which selected informative wavelengths by combining clustering concept and partial least square (PLS) methods to improve models’ performance by synchronous fluorescence spectra. The fluorescence spectra of tea samples were obtained and k-means and kohonen-self organizing map clustering algorithms were carried out to cluster full spectra into several clusters, and sub-PLS regression model was developed on each cluster. Finally, CL-PLS models consisting of gradually selected clusters were built. Correlation coefficient (R) was used to evaluate the effect on prediction performance of PLS models. In addition, variable influence on projection partial least square (VIP-PLS), selectivity ratio partial least square (SR-PLS), interval partial least square (iPLS) models and full spectra PLS model were investigated and the results were compared. The results showed that CL-PLS presented the best result for flavonoids prediction using synchronous fluorescence spectra.

Variable selection based on clustering analysis for improvement of polyphenols prediction in green tea using synchronous fluorescence spectra.

PubMed

Shan, Jiajia; Wang, Xue; Zhou, Hao; Han, Shuqing; Riza, Dimas Firmanda Al; Kondo, Naoshi

2018-03-13

Synchronous fluorescence spectra, combined with multivariate analysis were used to predict flavonoids content in green tea rapidly and nondestructively. This paper presented a new and efficient spectral intervals selection method called clustering based partial least square (CL-PLS), which selected informative wavelengths by combining clustering concept and partial least square (PLS) methods to improve models' performance by synchronous fluorescence spectra. The fluorescence spectra of tea samples were obtained and k-means and kohonen-self organizing map clustering algorithms were carried out to cluster full spectra into several clusters, and sub-PLS regression model was developed on each cluster. Finally, CL-PLS models consisting of gradually selected clusters were built. Correlation coefficient (R) was used to evaluate the effect on prediction performance of PLS models. In addition, variable influence on projection partial least square (VIP-PLS), selectivity ratio partial least square (SR-PLS), interval partial least square (iPLS) models and full spectra PLS model were investigated and the results were compared. The results showed that CL-PLS presented the best result for flavonoids prediction using synchronous fluorescence spectra.

Machine learning methods applied on dental fear and behavior management problems in children.

PubMed

Klingberg, G; Sillén, R; Norén, J G

1999-08-01

The etiologies of dental fear and dental behavior management problems in children were investigated in a database of information on 2,257 Swedish children 4-6 and 9-11 years old. The analyses were performed using computerized inductive techniques within the field of artificial intelligence. The database held information regarding dental fear levels and behavior management problems, which were defined as outcomes, i.e. dependent variables. The attributes, i.e. independent variables, included data on dental health and dental treatments, information about parental dental fear, general anxiety, socioeconomic variables, etc. The data contained both numerical and discrete variables. The analyses were performed using an inductive analysis program (XpertRule Analyser, Attar Software Ltd, Lancashire, UK) that presents the results in a hierarchic diagram called a knowledge tree. The importance of the different attributes is represented by their position in this diagram. The results show that inductive methods are well suited for analyzing multifactorial and complex relationships in large data sets, and are thus a useful complement to multivariate statistical techniques. The knowledge trees for the two outcomes, dental fear and behavior management problems, were very different from each other, suggesting that the two phenomena are not equivalent. Dental fear was found to be more related to non-dental variables, whereas dental behavior management problems seemed connected to dental variables.

A Tightly Coupled Non-Equilibrium Magneto-Hydrodynamic Model for Inductively Coupled RF Plasmas

DTIC Science & Technology

2016-02-29

development a tightly coupled magneto-hydrodynamic model for Inductively Coupled Radio- Frequency (RF) Plasmas. Non Local Thermodynamic Equilibrium (NLTE...for Inductively Coupled Radio-Frequency (RF) Plasmas. Non Local Thermodynamic Equilibrium (NLTE) effects are described based on a hybrid State-to-State... thermodynamic variable. This choice allows one to hide the non-linearity of the gas (total) thermal conductivity κ and can partially alle- 2 viate numerical

Transfer of benzo[a]pyrene from microplastics to Artemia nauplii and further to zebrafish via a trophic food web experiment: CYP1A induction and visual tracking of persistent organic pollutants.

PubMed

Batel, Annika; Linti, Frederic; Scherer, Martina; Erdinger, Lothar; Braunbeck, Thomas

2016-07-01

The uptake of microplastic particles and the transfer of potential harmful substances along with microplastics has been studied in a variety of organisms, especially invertebrates. However, the potential accumulation of very small microplastic particles along food webs ending with vertebrate models has not been investigated so far. Therefore, a simple artificial food chain with Artemia sp. nauplii and zebrafish (Danio rerio) was established to analyze the transfer of microplastic particles and associated persistent organic pollutants (POPs) between different trophic levels. Very small (1-20 μm) microplastic particles accumulated in Artemia nauplii and were subsequently transferred to fish. Virgin particles not loaded with POPs did not cause any observable physical harm in the intestinal tracts of zebrafish, although parts of the particles were retained within the mucus of intestinal villi and might even have been taken up by epithelial cells. The transfer of associated POPs was tested with the polycyclic aromatic hydrocarbon benzo[a]pyrene and an ethoxyresorufin-O-deethylase (EROD) assay for CYP1A induction in zebrafish liver as well as via fluorescence analyses. Whereas a significant induction in the EROD assay could not be shown, because of high individual variation and low sensitivity regarding substance concentration, the fluorescence tracking of benzo[a]pyrene indicates that food-borne microplastic-associated POPs may actually desorb in the intestine of fish and are thus transferred to the intestinal epithelium and liver. Environ Toxicol Chem 2016;35:1656-1666. © 2016 SETAC. © 2016 SETAC.

Myricetin and methyl eugenol combination enhances the anticancer activity, cell cycle arrest and apoptosis induction of cis-platin against HeLa cervical cancer cell lines

PubMed Central

Yi, Jin-Ling; Shi, Song; Shen, Yan-Li; Wang, Ling; Chen, Hai-Yan; Zhu, Jun; Ding, Yan

2015-01-01

Drug combination therapies are common practice in the treatment of cancer. In this study, we evaluated the anticancer effects of myricetin (MYR), methyl eugenol (MEG) and cisplatin (CP) both separately as well as in combination against cervical cancer (HeLa) cells. To demonstrate whether MYR and MEG enhance the anticancer activity of CP against cervical cancer cells, we treated HeLa cells with MYR and MEG alone or in combination with cisplatin and evaluated cell growth and apoptosis using MTT (3 (4, 5 dimethyl thiazol 2yl) 2, 5 diphenyltetrazolium bromide) assay, LDH release assay, flow cytometry and fluorescence microscopy. The results revealed that, as compared to single drug treatment, the combination of MYR or MEG with CP resulted in greater effect in inhibiting cancer cell growth and inducing apoptosis. Cell apoptosis induction, Caspase-3 activity, cell cycle arrest and mitochondrial membrane potential loss were systematically studied to reveal the mechanisms of synergy between MYR, MEG and CP. Combination of MYR or MEG with CP resulted in more potent apoptosis induction as revealed by fluorescence microscopy using Hoechst 33258 and AO-ETBR staining. The combination treatment also increased the number of cells in G0/G1 phase dramatically as compared to single drug treatment. Mitochondrial membrane potential loss (ΛΨm) as well as Caspase-3 activity was much higher in combination treatment as compared to single drug treatment. Findings of this investigation suggest that MYR and MEG combined with cisplatin is a potential clinical chemotherapeutic approach in human cervical cancer. PMID:25972998

Electron-Beam Diagnostic Methods for Hypersonic Flow Diagnostics

NASA Technical Reports Server (NTRS)

1994-01-01

The purpose of this work was the evaluation of the use of electron-bean fluorescence for flow measurements during hypersonic flight. Both analytical and numerical models were developed in this investigation to evaluate quantitatively flow field imaging concepts based upon the electron beam fluorescence technique for use in flight research and wind tunnel applications. Specific models were developed for: (1) fluorescence excitation/emission for nitrogen, (2) rotational fluorescence spectrum for nitrogen, (3) single and multiple scattering of electrons in a variable density medium, (4) spatial and spectral distribution of fluorescence, (5) measurement of rotational temperature and density, (6) optical filter design for fluorescence imaging, and (7) temperature accuracy and signal acquisition time requirements. Application of these models to a typical hypersonic wind tunnel flow is presented. In particular, the capability of simulating the fluorescence resulting from electron impact ionization in a variable density nitrogen or air flow provides the capability to evaluate the design of imaging instruments for flow field mapping. The result of this analysis is a recommendation that quantitative measurements of hypersonic flow fields using electron-bean fluorescence is a tractable method with electron beam energies of 100 keV. With lower electron energies, electron scattering increases with significant beam divergence which makes quantitative imaging difficult. The potential application of the analytical and numerical models developed in this work is in the design of a flow field imaging instrument for use in hypersonic wind tunnels or onboard a flight research vehicle.

Bioimaging of metallothioneins in ocular tissue sections by laser ablation-ICP-MS using bioconjugated gold nanoclusters as specific tags.

PubMed

Cruz-Alonso, María; Fernandez, Beatriz; Álvarez, Lydia; González-Iglesias, Héctor; Traub, Heike; Jakubowski, Norbert; Pereiro, Rosario

2017-12-18

An immunohistochemical method is described to visualize the distribution of metallothioneins 1/2 (MT 1/2) and metallothionein 3 (MT 3) in human ocular tissue. It is making use of (a) antibodies conjugated to gold nanoclusters (AuNCs) acting as labels, and (b) laser ablation (LA) coupled to inductively coupled plasma - mass spectrometry (ICP-MS). Water-soluble fluorescent AuNCs (with an average size of 2.7 nm) were synthesized and then conjugated to antibody by carbodiimide coupling. The surface of the modified AuNCs was then blocked with hydroxylamine to avoid nonspecific interactions with biological tissue. Immunoassays for MT 1/2 and MT 3 in ocular tissue sections (5 μm thick) from two post mortem human donors were performed. Imaging studies were then performed by fluorescence using confocal microscopy, and LA-ICP-MS was performed in the retina to measure the signal for gold. Signal amplification by the >500 gold atoms in each nanocluster allowed the antigens (MT 1/2 and MT 3) to be imaged by LA-ICP-MS using a laser spot size as small as 4 μm. The image patterns found in retina are in good agreement with those obtained by conventional fluorescence immunohistochemistry which was used as an established reference method. Graphical abstract Gold nanoclusters (AuNCs) conjugated to a primary specific antibody serve as a label for amplified bioimaging of metallothioneins (MTs) by laser ablation coupled to inductively coupled plasma - mass spectrometry (ICP-MS) in human ocular tissue sections.

Laser microbeams for DNA damage induction, optical tweezers for the search on blood pressure relaxing drugs: contributions to ageing research

NASA Astrophysics Data System (ADS)

Grigaravicius, P.; Monajembashi, S.; Hoffmann, M.; Altenberg, B.; Greulich, K. O.

2009-08-01

One essential cause of human ageing is the accumulation of DNA damages during lifetime. Experimental studies require quantitative induction of damages and techniques to visualize the subsequent DNA repair. A new technique, the "immuno fluorescent comet assay", is used to directly visualize DNA damages in the microscope. Using DNA repair proteins fluorescently labeled with green fluorescent protein, it could be shown that the repair of the most dangerous DNA double strand breaks starts with the inaccurate "non homologous end joining" pathway and only after 1 - 1 ½ minutes may switch to the more accurate "homologous recombination repair". One might suggest investigating whether centenarians use "homologous recombination repair" differently from those ageing at earlier years and speculate whether it is possible, for example by nutrition, to shift DNA repair to a better use of the error free pathway and thus promote healthy ageing. As a complementary technique optical tweezers, and particularly its variant "erythrocyte mediated force application", is used to simulate the effects of blood pressure on HUVEC cells representing the inner lining of human blood vessels. Stimulating one cell induces in the whole neighbourhood waves of calcium and nitric oxide, known to relax blood vessels. NIFEDIPINE and AMLODIPINE, both used as drugs in the therapy of high blood pressure, primarily a disease of the elderly, prolong the availability of nitric oxide. This partially explains their mode of action. In contrast, VERAPAMILE, also a blood pressure reducing drug, does not show this effect, indicating that obviously an alternative mechanism must be responsible for vessel relaxation.

A chromosomally encoded T7 RNA polymerase-dependent gene expression system for Corynebacterium glutamicum: construction and comparative evaluation at the single-cell level

PubMed Central

Kortmann, Maike; Kuhl, Vanessa; Klaffl, Simon; Bott, Michael

2015-01-01

Corynebacterium glutamicum has become a favourite model organism in white biotechnology. Nevertheless, only few systems for the regulatable (over)expression of homologous and heterologous genes are currently available, all of which are based on the endogenous RNA polymerase. In this study, we developed an isopropyl-β-d-1-thiogalactopyranosid (IPTG)-inducible T7 expression system in the prophage-free strain C. glutamicum MB001. For this purpose, part of the DE3 region of Escherichia coli BL21(DE3) including the T7 RNA polymerase gene 1 under control of the lacUV5 promoter was integrated into the chromosome, resulting in strain MB001(DE3). Furthermore, the expression vector pMKEx2 was constructed allowing cloning of target genes under the control of the T7lac promoter. The properties of the system were evaluated using eyfp as heterologous target gene. Without induction, the system was tightly repressed, resulting in a very low specific eYFP fluorescence (= fluorescence per cell density). After maximal induction with IPTG, the specific fluorescence increased 450-fold compared with the uninduced state and was about 3.5 times higher than in control strains expressing eyfp under control of the IPTG-induced tac promoter with the endogenous RNA polymerase. Flow cytometry revealed that T7-based eyfp expression resulted in a highly uniform population, with 99% of all cells showing high fluorescence. Besides eyfp, the functionality of the corynebacterial T7 expression system was also successfully demonstrated by overexpression of the C. glutamicum pyk gene for pyruvate kinase, which led to an increase of the specific activity from 2.6 to 135 U mg−1. It thus presents an efficient new tool for protein overproduction, metabolic engineering and synthetic biology approaches with C. glutamicum. PMID:25488698

Diurnal Variability in Chlorophyll-a, Carotenoids, CDOM and SO₄(2-) Intensity of Offshore Seawater Detected by an Underwater Fluorescence-Raman Spectral System.

PubMed

Chen, Jing; Ye, Wangquan; Guo, Jinjia; Luo, Zhao; Li, Ying

2016-07-13

A newly developed integrated fluorescence-Raman spectral system (λex = 532 nm) for detecting Chlorophyll-a (chl-a), Chromophoric Dissolved Organic Matter (CDOM), carotenoids and SO₄(2-) in situ was used to successfully investigate the diurnal variability of all above. Simultaneously using the integration of fluorescence spectroscopy and Raman spectroscopy techniques provided comprehensive marine information due to the complementarity between the different excitation mechanisms and different selection rules. The investigation took place in offshore seawater of the Yellow Sea (36°05'40'' N, 120°31'32'' E) in October 2014. To detect chl-a, CDOM, carotenoids and SO₄(2-), the fluorescence-Raman spectral system was deployed. It was found that troughs of chl-a and CDOM fluorescence signal intensity were observed during high tides, while the signal intensity showed high values with larger fluctuations during ebb-tide. Chl-a and carotenoids were influenced by solar radiation within a day cycle by different detection techniques, as well as displaying similar and synchronous tendency. CDOM fluorescence cause interference to the measurement of SO₄(2-). To avoid such interference, the backup Raman spectroscopy system with λex = 785 nm was employed to detect SO₄(2-) concentration on the following day. The results demonstrated that the fluorescence-Raman spectral system has great potential in detection of chl-a, carotenoids, CDOM and SO₄(2-) in the ocean.

Aptamer-Based Dual-Functional Probe for Rapid and Specific Counting and Imaging of MCF-7 Cells.

PubMed

Yang, Bin; Chen, Beibei; He, Man; Yin, Xiao; Xu, Chi; Hu, Bin

2018-02-06

Development of multimodal detection technologies for accurate diagnosis of cancer at early stages is in great demand. In this work, we report a novel approach using an aptamer-based dual-functional probe for rapid, sensitive, and specific counting and visualization of MCF-7 cells by inductively coupled plasma-mass spectrometry (ICP-MS) and fluorescence imaging. The probe consists of a recognition unit of aptamer to catch cancer cells specifically, a fluorescent dye (FAM) moiety for fluorescence resonance energy transfer (FRET)-based "off-on" fluorescence imaging as well as gold nanoparticles (Au NPs) tag for both ICP-MS quantification and fluorescence quenching. Due to the signal amplification effect and low spectral interference of Au NPs in ICP-MS, an excellent linearity and sensitivity were achieved. Accordingly, a limit of detection of 81 MCF-7 cells and a relative standard deviation of 5.6% (800 cells, n = 7) were obtained. The dynamic linear range was 2 × 10 2 to 1.2 × 10 4 cells, and the recoveries in human whole blood were in the range of 98-110%. Overall, the established method provides quantitative and visualized information on MCF-7 cells with a simple and rapid process and paves the way for a promising strategy for biomedical research and clinical diagnostics.

Field oriented control of induction motors

NASA Technical Reports Server (NTRS)

Burrows, Linda M.; Zinger, Don S.; Roth, Mary Ellen

1990-01-01

Induction motors have always been known for their simple rugged construction, but until lately were not suitable for variable speed or servo drives due to the inherent complexity of the controls. With the advent of field oriented control (FOC), however, the induction motor has become an attractive option for these types of drive systems. An FOC system which utilizes the pulse population modulation method to synthesize the motor drive frequencies is examined. This system allows for a variable voltage to frequency ratio and enables the user to have independent control of both the speed and torque of an induction motor. A second generation of the control boards were developed and tested with the next point of focus being the minimization of the size and complexity of these controls. Many options were considered with the best approach being the use of a digital signal processor (DSP) due to its inherent ability to quickly evaluate control algorithms. The present test results of the system and the status of the optimization process using a DSP are discussed.

Significance of Cytochrome P450 System Responses and Levels of Bile Fluorescent Aromatic Compounds in Marine Wildlife Following Oil Spills

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

Lee, Richard F.; Anderson, Jack W.

2005-07-01

The relationships among cytochrome P450 induction in marine wildlife species, levels of fluorescent aromatic compounds (FAC) in their bile, the chemical composition of the inducing compounds, the significance of the exposure pathway, and any resulting injury, as a consequence of exposure to crude oil following a spill, are reviewed. Fish collected after oil spills often show increases in cytochrome P450 system activity, cytochrome P4501A (CYP1A) and bile fluorescent aromatic compounds (FAC), that are correlated with exposure to polycyclic aromatic hydrocarbons (PAH) in the oil. There is also some evidence for increases in bile FAC and induction of cytochrome P450 inmore » marine birds and mammals after oil spills. However, when observed, increases in these exposure indicators are transitory and generally decrease to background levels within one year after the exposure. Laboratory studies have shown induction of cytochrome P450 systems occurs after exposure of fish to crude oil in water, sediment or food. Most of the PAH found in crude oil (dominantly 2- and 3-ring PAH) are not strong inducers of cytochrome P450. Exposure to the 4-ring chrysenes or the photooxidized products of the PAH may account for the cytochrome P450 responses in fish collected from oil-spill sites. The contribution of non-spill background PAH, particularly combustion-derived (pyrogenic) PAH, to bile FAC and cytochrome P450 system responses can be confounding and needs to be considered when evaluating oil spill effects. The ubiquity of pyrogenic PAH makes it important to fully characterize all sources of PAH, including PAH from natural resources, e.g. retene, in oil spill studies. In addition, such parameters as species, sex, age, ambient temperature and season need to be taken into account. While increases in fish bile FAC and cytochrome P450 system responses, can together, be sensitive general indicators of PAH exposure after an oil spill, there is little unequivocal evidence to suggest a linkage to higher order biological effects, e.g. toxicity, lesions, reproductive failure.« less

Enhanced resistance to citrus canker in transgenic mandarin expressing Xa21 from rice.

PubMed

Omar, Ahmad A; Murata, Mayara M; El-Shamy, Hesham A; Graham, James H; Grosser, Jude W

2018-04-01

Genetic engineering approaches offer an alternative method to the conventional breeding of Citrus sp. 'W. Murcott' mandarin (a hybrid of 'Murcott' and an unknown pollen parent) is one of the most commercially important cultivars grown in many regions around the world. Transformation of 'W. Murcott' mandarin was achieved by direct DNA uptake using a protoplast transformation system. DNA construct (pAO3), encoding Green Fluorescent Protein (GFP) and the cDNA of Xa21, a Xanthomonas resistance gene from rice, was used to transform protoplasts of 'W. Murcott' mandarin. Following citrus protoplast culture and regeneration, transformed micro calli were microscopically designated via GFP expression, physically isolated from non-transformed tissue, and cultured on somatic embryogenesis induction medium. More than 150 transgenic embryos were recovered and from them, ten transgenic lines were regenerated and cultured on rooting medium for shoot elongation. Transgenic shoots were micrografted and established in the greenhouse with 3-5 replicates per line. The insertion of Xa21 and GFP was confirmed by PCR and southern blot analysis. GFP expression was verified by fluorescence microscopy and western blot analysis revealed expression of Xa21 although it was variable among transgenic lines, as shown by RT-qPCR. Transgenic plants challenged with the citrus canker pathogen by syringe inoculation showed a reduction in lesion number and bacterial populations within lesions compared to non-transgenic control plants. Transgenic 'W. Murcott' mandarin lines with improved canker resistance via protoplast transformation from embryogenic callus with the Xa21 gene from rice are being evaluated under field conditions to validate the level of resistance.

Effect of a Short-term Fast on Ketamine–Xylazine Anesthesia in Rats

PubMed Central

Struck, Maggie B; Andrutis, Karl A; Ramirez, Harvey E; Battles, August H

2011-01-01

Although ketamine–xylazine (KX) anesthesia is commonly used in rats, it is often reported to have an inconsistent anesthetic effect, with a prolonged induction time, an inadequate anesthetic plane, or a very short sleep time. Blood flow to the liver is known to shift after a meal in rats, perhaps explaining anesthetic variability among rats with variable prandial status. The current study tested the hypothesis that a short period of fasting (3 h) prior to induction with intraperitoneal KX anesthesia would provide a shorter time to recumbency, a longer total sleep time, and a more consistent loss of toe pinch response than would fed rats. Two groups of male Sprague–Dawley rats were used in blinded, crossover experiments. KX anesthesia was administered at 2 different doses (50 mg/kg–5 mg/kg and 70 mg/kg–7 mg/kg) after ad libitum feeding or a 3-h fast. There were no significant differences between groups in induction time, total sleep time, or loss of toe pinch response. We conclude that fasting rats for 3 h prior to KX intraperitoneal anesthesia does not affect induction time, total sleep time, loss of toe pinch response or reduce KX anesthetic variability in male Sprague–Dawley rats. PMID:21640029

Processes Affecting the Variability of Fluorescence Signals from Benthic Targets in Shallow Waters

DTIC Science & Technology

1998-09-30

analysis of zooxanthellae isolated from coral samples. We examined fluorescence lifetimes from benthic targets using femtosecond laser based Single...a temporal resolution 5 ps. The laser set-up was employed for laboratory studies of fluorescence lifetimes from model targets such as zooxanthellae ...seagrasses. Over 200 measurements on isolated zooxanthellae were conducted with a bench-top FRR fluorometer and Phase Shift Fluorometer. During the CoBOP-98

Parametric models to compute tryptophan fluorescence wavelengths from classical protein simulations.

PubMed

Lopez, Alvaro J; Martínez, Leandro

2018-02-26

Fluorescence spectroscopy is an important method to study protein conformational dynamics and solvation structures. Tryptophan (Trp) residues are the most important and practical intrinsic probes for protein fluorescence due to the variability of their fluorescence wavelengths: Trp residues emit in wavelengths ranging from 308 to 360 nm depending on the local molecular environment. Fluorescence involves electronic transitions, thus its computational modeling is a challenging task. We show that it is possible to predict the wavelength of emission of a Trp residue from classical molecular dynamics simulations by computing the solvent-accessible surface area or the electrostatic interaction between the indole group and the rest of the system. Linear parametric models are obtained to predict the maximum emission wavelengths with standard errors of the order 5 nm. In a set of 19 proteins with emission wavelengths ranging from 308 to 352 nm, the best model predicts the maximum wavelength of emission with a standard error of 4.89 nm and a quadratic Pearson correlation coefficient of 0.81. These models can be used for the interpretation of fluorescence spectra of proteins with multiple Trp residues, or for which local Trp environmental variability exists and can be probed by classical molecular dynamics simulations. © 2018 Wiley Periodicals, Inc. © 2018 Wiley Periodicals, Inc.

Fluorescence-Activated Cell Sorting-Based Analysis Reveals an Asymmetric Induction of Interferon-Stimulated Genes in Response to Seasonal Influenza A Virus

PubMed Central

von Recum-Knepper, Jessica; Sadewasser, Anne; Weinheimer, Viola K.

2015-01-01

ABSTRACT Influenza A virus (IAV) infection provokes an antiviral response involving the expression of type I and III interferons (IFN) and IFN-stimulated genes (ISGs) in infected cell cultures. However, the spatiotemporal dynamics of the IFN reaction are incompletely understood, as previous studies investigated mainly the population responses of virus-infected cultures, although substantial cell-to-cell variability has been documented. We devised a fluorescence-activated cell sorting-based assay to simultaneously quantify expression of viral antigens and ISGs, such as ISG15, MxA, and IFIT1, in IAV-infected cell cultures at the single-cell level. This approach revealed that seasonal IAV triggers an unexpected asymmetric response, as the major cell populations expressed either viral antigen or ISG, but rarely both. Further investigations identified a role of the viral NS1 protein in blocking ISG expression in infected cells, which surprisingly did not reduce paracrine IFN signaling to noninfected cells. Interestingly, viral ISG control was impaired in cultures infected with avian-origin IAV, including the H7N9 virus from eastern China. This phenotype was traced back to polymorphic NS1 amino acids known to be important for stable binding of the polyadenylation factor CPSF30 and concomitant suppression of host cell gene expression. Most significantly, mutation of two amino acids within the CPSF30 attachment site of NS1 from seasonal IAV diminished the strict control of ISG expression in infected cells and substantially attenuated virus replication. In conclusion, our approach revealed an asymmetric, NS1-dependent ISG induction in cultures infected with seasonal IAV, which appears to be essential for efficient virus propagation. IMPORTANCE Interferons are expressed by infected cells in response to IAV infection and play important roles in the antiviral immune response by inducing hundreds of interferon-stimulated genes (ISGs). Unlike many previous studies, we investigated the ISG response at the single-cell level, enabling novel insights into this virus-host interaction. Hence, cell cultures infected with seasonal IAV displayed an asymmetric ISG induction that was confined almost exclusively to noninfected cells. In comparison, ISG expression was observed in larger cell populations infected with avian-origin IAV, suggesting a more resolute antiviral response to these strains. Strict control of ISG expression by seasonal IAV was explained by the binding of the viral NS1 protein to the polyadenylation factor CPSF30, which reduces host cell gene expression. Mutational disruption of CPSF30 binding within NS1 concomitantly attenuated ISG control and replication of seasonal IAV, illustrating the importance of maintaining an asymmetric ISG response for efficient virus propagation. PMID:25903337

Cytotoxic effects of Cochlospermum regium (Mart & Schrank) Pilger aqueous root extract on mammalian cells.

PubMed

Ceschini, Livônios; Campos, Elida Geralda

2006-01-16

We investigated the effect of Cochlospermum regium (Mart & Schrank) Pilger aqueous root extract on Chinese hamster ovarian (CHO)-K1 cells. The extract significantly decreased proliferation of CHO-K1 cells (EC(50)=1.5mg/mL). Apoptosis induction was analysed by fluorescent microscopy. Cell cultures treated with Cochlospermum regium extract for 4h contained 13.6% apoptotic cells after 24h (investigated by fluorescent DNA-microscopy with acridine orange/ethidium bromide staining). Characteristic chromatin condensation and fragmentation, verified by 4',6-diamidino-2-phenylindole (DAPI) staining, was observed in the cells after treatment with Cochlospermum regium extract. The results confirm the toxicity of Cochlospermum regium root extract to immortal, non-tumorigenic mammalian cells in vitro.

Silica nanoparticle-based dual imaging colloidal hybrids: cancer cell imaging and biodistribution

PubMed Central

Lee, Haisung; Sung, Dongkyung; Kim, Jinhoon; Kim, Byung-Tae; Wang, Tuntun; An, Seong Soo A; Seo, Soo-Won; Yi, Dong Kee

2015-01-01

In this study, fluorescent dye-conjugated magnetic resonance (MR) imaging agents were investigated in T mode. Gadolinium-conjugated silica nanoparticles were successfully synthesized for both MR imaging and fluorescence diagnostics. Polyamine and polycarboxyl functional groups were modified chemically on the surface of the silica nanoparticles for efficient conjugation of gadolinium ions. The derived gadolinium-conjugated silica nanoparticles were investigated by zeta potential analysis, transmission electron microscopy, inductively coupled plasma mass spectrometry, and energy dispersive x-ray spectroscopy. MR equipment was used to investigate their use as contrast-enhancing agents in T1 mode under a 9.4 T magnetic field. In addition, we tracked the distribution of the gadolinium-conjugated nanoparticles in both lung cancer cells and organs in mice. PMID:26357472

Minimally-invasive Laser Ablation Inductively Coupled Plasma Mass Spectrometry analysis of model ancient copper alloys

NASA Astrophysics Data System (ADS)

Walaszek, Damian; Senn, Marianne; Wichser, Adrian; Faller, Markus; Wagner, Barbara; Bulska, Ewa; Ulrich, Andrea

2014-09-01

This work describes an evaluation of a strategy for multi-elemental analysis of typical ancient bronzes (copper, lead bronze and tin bronze) by means of laser ablation inductively coupled plasma mass spectrometry (LA-ICPMS).The samples originating from archeological experiments on ancient metal smelting processes using direct reduction in a ‘bloomery’ furnace as well as historical casting techniques were investigated with the use of the previously proposed analytical procedure, including metallurgical observation and preliminary visual estimation of the homogeneity of the samples. The results of LA-ICPMS analysis were compared to the results of bulk composition obtained by X-ray fluorescence spectrometry (XRF) and by inductively coupled plasma mass spectrometry (ICPMS) after acid digestion. These results were coherent for most of the elements confirming the usefulness of the proposed analytical procedure, however the reliability of the quantitative information about the content of the most heterogeneously distributed elements was also discussed in more detail.

Development of cytochromes P450 in avian species as a biomarker for environmental contaminant exposure and effect: Procedures and baseline values

USGS Publications Warehouse

Melancon, M.J.; Bengston, David A.; Henshel, Diane S.

1996-01-01

As in mammals and fish, birds respond to many environmental contaminants with induction of hepatic cytochromes P450. In order to monitor cytchromes P450 in specific avian species, for assessing the status of that species or the habitat it utilizes, it is necessary to have background information on the appropriate assay conditions and the responsiveness of cytochrome P450 induction in that species. Assay of four monooxygenases which give resorufin as product using a fluorescence microwell plate scanner has proven to be an effective approach. Information is provided on the incubation conditions and baseline activity for twenty avian species at ages ranging from pipping embryo to adult. Induction responsiveness is presented for sixteen of them. This information can serve as a guide for those who wish to utilize cytochrome P450 as a biomarker for contaminant exposure and effect to aid in selection of appropriate species, age, and monooxygenase assay(s).

Dimmable electronic ballasts by variable power density modulation technique

NASA Astrophysics Data System (ADS)

Borekci, Selim; Kesler, Selami

2014-11-01

Dimming can be accomplished commonly by switching frequency and pulse density modulation techniques and a variable inductor. In this study, a variable power density modulation (VPDM) control technique is proposed for dimming applications. A fluorescent lamp is operated in several states to meet the desired lamp power in a modulation period. The proposed technique has the same advantages of magnetic dimming topologies have. In addition, a unique and flexible control technique can be achieved. A prototype dimmable electronic ballast is built and experiments related to it have been conducted. As a result, a 36WT8 fluorescent lamp can be driven for a desired lamp power from several alternatives without modulating the switching frequency.

Use of Complementary Approaches to Imaging Biomolecules and Endogenous and Exogenous Trace Elements and Nanoparticles in Biological Samples

NASA Astrophysics Data System (ADS)

Brown, Koshonna Dinettia

X-ray Fluorescence Microscopy (XFM) is a useful technique for study of biological samples. XFM was used to map and quantify endogenous biological elements as well as exogenous materials in biological samples, such as the distribution of titanium dioxide (TiO2) nanoparticles. TiO 2 nanoparticles are produced for many different purposes, including development of therapeutic and diagnostic particles for cancer detection and treatment, drug delivery, and induction of DNA breaks. Delivery of such nanoparticles can be targeted to specific cells and subcellular structures. In this work, we develop two novel approaches to stain TiO2 nanoparticles for optical microscopy and to confirm that staining by XFM. The first approach utilizes fluorescent biotin and fluorescent streptavidin to label the nanoparticles before and after cellular uptake; the second approach is based on the copper-catalyzed azide-alkyne cycloaddition, the so-called CLICK chemistry, for labeling of azide conjugated TiO2 nanoparticles with "clickable" dyes such as alkyne Alexa Fluor dyes with a high fluorescent yield. To confirm that the optical fluorescence signals of nanoparticles stained in situ match the distribution of the Ti element, we used high resolution synchrotron X-Ray Fluorescence Microscopy (XFM) using the Bionanoprobe instrument at the Advanced Photon Source at Argonne National Laboratory. Titanium-specific X-ray fluorescence showed excellent overlap with the location of Alexa Fluor optical fluorescence detected by confocal microscopy. In this work XFM was also used to investigate native elemental differences between two different types of head and neck cancer, one associated with human papilloma virus infection, the other virus free. Future work may see a cross between these themes, for example, exploration of TiO2 nanoparticles as anticancer treatment for these two different types of head and neck cancer.

Topography of Cells Revealed by Variable-Angle Total Internal Reflection Fluorescence Microscopy.

PubMed

Cardoso Dos Santos, Marcelina; Déturche, Régis; Vézy, Cyrille; Jaffiol, Rodolphe

2016-09-20

We propose an improved version of variable-angle total internal reflection fluorescence microscopy (vaTIRFM) adapted to modern TIRF setup. This technique involves the recording of a stack of TIRF images, by gradually increasing the incident angle of the light beam on the sample. A comprehensive theory was developed to extract the membrane/substrate separation distance from fluorescently labeled cell membranes. A straightforward image processing was then established to compute the topography of cells with a nanometric axial resolution, typically 10-20 nm. To highlight the new opportunities offered by vaTIRFM to quantify adhesion process of motile cells, adhesion of MDA-MB-231 cancer cells on glass substrate coated with fibronectin was examined. Copyright © 2016 Biophysical Society. Published by Elsevier Inc. All rights reserved.

Development of tolerance of egg plant (Solanum melangena L.) to field application of dimethoate.

PubMed

Khillar, R; Acharya, S; Mohapatra, P K

2010-07-01

Dimethoate, at field concentration (1.419 mg g(-1) fr wt), caused inhibition of photosynthesis, transpiration and stomatal conductance of Solanum melangena L. on first treatment but subsequent treatments caused adaptation and recovery of these parameters. The variable fluorescence (F(v)), dissipation (DI(0)/RC), 2 ms relative variable fluorescence (V(j)), net rate of PS II closure (M(0)), and maximum trapping rate of active PS II (TR(0)/RC) increased initially but reduced to the control value with repetition of treatment. However, fluorescence yield (TR(0)/Abs), electron transport probability (ET(0)/TR(0)) and activity of RC (ET(0)/RC) increased with each treatment. With each subsequent treatment there was enhancement of activities of esterases and decrease of insecticide content of leaves.

Unmixing of fluorescence spectra to resolve quantitative time-series measurements of gene expression in plate readers.

PubMed

Lichten, Catherine A; White, Rachel; Clark, Ivan B N; Swain, Peter S

2014-02-03

To connect gene expression with cellular physiology, we need to follow levels of proteins over time. Experiments typically use variants of Green Fluorescent Protein (GFP), and time-series measurements require specialist expertise if single cells are to be followed. Fluorescence plate readers, however, a standard in many laboratories, can in principle provide similar data, albeit at a mean, population level. Nevertheless, extracting the average fluorescence per cell is challenging because autofluorescence can be substantial. Here we propose a general method for correcting plate reader measurements of fluorescent proteins that uses spectral unmixing and determines both the fluorescence per cell and the errors on that fluorescence. Combined with strain collections, such as the GFP fusion collection for budding yeast, our methodology allows quantitative measurements of protein levels of up to hundreds of genes and therefore provides complementary data to high throughput studies of transcription. We illustrate the method by following the induction of the GAL genes in Saccharomyces cerevisiae for over 20 hours in different sugars and argue that the order of appearance of the Leloir enzymes may be to reduce build-up of the toxic intermediate galactose-1-phosphate. Further, we quantify protein levels of over 40 genes, again over 20 hours, after cells experience a change in carbon source (from glycerol to glucose). Our methodology is sensitive, scalable, and should be applicable to other organisms. By allowing quantitative measurements on a per cell basis over tens of hours and over hundreds of genes, it should increase our understanding of the dynamic changes that drive cellular behaviour.

Unmixing of fluorescence spectra to resolve quantitative time-series measurements of gene expression in plate readers

PubMed Central

2014-01-01

Background To connect gene expression with cellular physiology, we need to follow levels of proteins over time. Experiments typically use variants of Green Fluorescent Protein (GFP), and time-series measurements require specialist expertise if single cells are to be followed. Fluorescence plate readers, however, a standard in many laboratories, can in principle provide similar data, albeit at a mean, population level. Nevertheless, extracting the average fluorescence per cell is challenging because autofluorescence can be substantial. Results Here we propose a general method for correcting plate reader measurements of fluorescent proteins that uses spectral unmixing and determines both the fluorescence per cell and the errors on that fluorescence. Combined with strain collections, such as the GFP fusion collection for budding yeast, our methodology allows quantitative measurements of protein levels of up to hundreds of genes and therefore provides complementary data to high throughput studies of transcription. We illustrate the method by following the induction of the GAL genes in Saccharomyces cerevisiae for over 20 hours in different sugars and argue that the order of appearance of the Leloir enzymes may be to reduce build-up of the toxic intermediate galactose-1-phosphate. Further, we quantify protein levels of over 40 genes, again over 20 hours, after cells experience a change in carbon source (from glycerol to glucose). Conclusions Our methodology is sensitive, scalable, and should be applicable to other organisms. By allowing quantitative measurements on a per cell basis over tens of hours and over hundreds of genes, it should increase our understanding of the dynamic changes that drive cellular behaviour. PMID:24495318

Reinforcement and Induction of Operant Variability

ERIC Educational Resources Information Center

Neuringer, Allen

2012-01-01

The target paper by Barba (2012) raises issues that were the focus of the author's first two publications on operant variability. The author will describe the main findings in those papers and then discuss Barba's specific arguments. Barba has argued against the operant nature of variability. (Contains 2 figures.)

SELECTIVITY AND SPECIFICITY OF SMALL MOLECULE FLUORESCENT DYES/PROBES USED FOR THE DETECTION OF Zn2+ AND Ca2+ IN CELLS

PubMed Central

Landero-Figueroa, Julio A.; Vignesh, Kavitha Subramanian; Deepe, George; Caruso, Joseph

2014-01-01

Fluorescent dyes are widely used in the detection of labile (free or exchangeable) Zn2+ and Ca2+ in living cells. However, their specificity over other cations and selectivity for detection of labile vs. protein-bound metal in cells remains unclear. We characterized these important properties for commonly used Zn2+ and Ca2+ dyes in a cellular environment. By tracing the fluorescence emission signal along with UV-Vis and size exclusion chromatography-inductively coupled plasma mass spectrometry (SEC-ICP-MS) in tandem, we demonstrated that among the dyes used for Zn2+, Zinpyr-1 fluoresces in the low molecular mass (LMM) region containing labile Zn2+, but also fluoresces in different molecular mass regions where zinc ion is detected. However, FluoZin™-3 AM, Newport Green™ DCF and Zinquin ethyl ester display weak fluorescence, lack of metal specificity and respond strongly in the high molecular mass (HMM) region. Four Ca2+ dyes were studied in an unperturbed cellular environment, and two of these were tested for binding behavior under an intracellular Ca2+ release stimulus. A majority of Ca2+ was in the labile form as tested by SEC-ICP-MS, but the fluorescence from Calcium Green-1™ AM, Oregon Green® 488 BAPTA-1, Fura red™ AM and Fluo-4 NW dyes in cells did not correspond to free Ca2+ detection. Instead, the dyes showed non-specific fluorescence in the mid- and high-molecular mass regions containing Zn, Fe and Cu. Proteomic analysis of one of the commonly seen fluorescing regions showed the possibility for some dyes to recognize Zn and Cu bound to metallothionein-2. These studies indicate that Zn2+ and Ca2+ binding dyes manifest fluorescence responses that are not unique to recognition of labile metals and bind other metals, leading to suboptimal specificity and selectivity. PMID:24356796

Fifteen years of PARAFAC application to organic matter fluorescence - progress, problems and possibilities

NASA Astrophysics Data System (ADS)

Murphy, K.; Stedmon, C. A.; Wunsch, U.

2017-12-01

The study of dissolved organic matter in aquatic milieu frequently involves measuring and interpreting fluorescence excitation emission matrices (EEMs) as a proxy for studying the total organic matter pool. Parallel Factor Analysis (PARAFAC) is used widely to identify and track independent organic matter fractions. This approach assumes that each EEM reflects the combined fluorescence signal from a limited number of unique, non-interacting chemical components, which are determined via a fitting algorithm. During the past fifteen years, considerable progress in understanding dissolved organic matter fluorescence has been achieved with the aid of PARAFAC; however, very few identical or ubiquitous fluorescence spectra have been independently identified. We studied the influence of wavelength selection on PARAFAC models and found this factor to have a decisive impact on PARAFAC spectra despite receiving little attention in most studies. Because large, chemically-diverse datasets may be too complex to analyse with PARAFAC, we are exploring novel methods for increasing variability in small datasets in order to reduce biases and increase interpretability. Our results suggest that spectral variability in PARAFAC models between studies are in many cases due to artefacts that could be minimised by careful experimental and modelling approaches.

Ratiometric analysis of Acridine Orange staining in the study of acidic organelles and autophagy.

PubMed

Thomé, Marcos P; Filippi-Chiela, Eduardo C; Villodre, Emilly S; Migliavaca, Celina B; Onzi, Giovana R; Felipe, Karina B; Lenz, Guido

2016-12-15

Acridine Orange is a cell-permeable green fluorophore that can be protonated and trapped in acidic vesicular organelles (AVOs). Its metachromatic shift to red fluorescence is concentration-dependent and, therefore, Acridine Orange fluoresces red in AVOs, such as autolysosomes. This makes Acridine Orange staining a quick, accessible and reliable method to assess the volume of AVOs, which increases upon autophagy induction. Here, we describe a ratiometric analysis of autophagy using Acridine Orange, considering the red-to-green fluorescence intensity ratio (R/GFIR) to quantify flow cytometry and fluorescence microscopy data of Acridine-Orange-stained cells. This method measured with accuracy the increase in autophagy induced by starvation or rapamycin, and the reduction in autophagy produced by bafilomycin A1 or the knockdown of Beclin1 or ATG7. Results obtained with Acridine Orange, considering R/GFIR, correlated with the conversion of the unlipidated form of LC3 (LC3-I) into the lipidated form (LC3-II), SQSTM1 degradation and GFP-LC3 puncta formation, thus validating this assay to be used as an initial and quantitative method for evaluating the late step of autophagy in individual cells, complementing other methods. © 2016. Published by The Company of Biologists Ltd.

Observation of aggregation triggered by Resonance Energy Transfer (RET) induced intermolecular pairing force.

PubMed

Pan, Xiaoyong; Wang, Weizhi; Ke, Lin; Zhang, Nan

2017-07-20

In this report, we showed the existence of RET induced intermolecular pairing force by comparing their fluorescence behaviors under room illumination vs standing in dark area for either PFluAnt solution or PFluAnt&PFOBT mixture. Their prominent emission attenuation under room illumination brought out the critical role of photo, i.e. RET induced intermolecular pairing force in induction of polymer aggregation. Constant UV-Vis absorption and fluorescence spectra in terms of both peak shapes and maximum wavelengths implied no chemical decomposition was involved. Recoverable fluorescence intensity, fluorescence lifetime as well as NMR spectra further exclude photo induced decomposition. The controllable on/off state of RET induced intermolecular pairing force was verified by the masking effect of outside PFluAnt solution which function as filter to block the excitation of inside PFluAnt and thus off the RET induced intermolecular pairing force. Theoretical calculation suggest that magnitude of RET induced intermolecular pairing force is on the same scale as that of van der Waals interaction. Although the absolute magnitude of RET induced intermolecular pairing force was not tunable, its effect can be magnified by intentionally turn it "on", which was achieved by irradiance with 5 W desk lamp in this report.

Diurnal Variability in Chlorophyll-a, Carotenoids, CDOM and SO42− Intensity of Offshore Seawater Detected by an Underwater Fluorescence-Raman Spectral System

PubMed Central

Chen, Jing; Ye, Wangquan; Guo, Jinjia; Luo, Zhao; Li, Ying

2016-01-01

A newly developed integrated fluorescence-Raman spectral system (λex = 532 nm) for detecting Chlorophyll-a (chl-a), Chromophoric Dissolved Organic Matter (CDOM), carotenoids and SO42− in situ was used to successfully investigate the diurnal variability of all above. Simultaneously using the integration of fluorescence spectroscopy and Raman spectroscopy techniques provided comprehensive marine information due to the complementarity between the different excitation mechanisms and different selection rules. The investigation took place in offshore seawater of the Yellow Sea (36°05′40′′ N, 120°31′32′′ E) in October 2014. To detect chl-a, CDOM, carotenoids and SO42−, the fluorescence-Raman spectral system was deployed. It was found that troughs of chl-a and CDOM fluorescence signal intensity were observed during high tides, while the signal intensity showed high values with larger fluctuations during ebb-tide. Chl-a and carotenoids were influenced by solar radiation within a day cycle by different detection techniques, as well as displaying similar and synchronous tendency. CDOM fluorescence cause interference to the measurement of SO42−. To avoid such interference, the backup Raman spectroscopy system with λex = 785 nm was employed to detect SO42− concentration on the following day. The results demonstrated that the fluorescence-Raman spectral system has great potential in detection of chl-a, carotenoids, CDOM and SO42− in the ocean. PMID:27420071

Primary production estimates from recordings of solar-stimulated fluorescence in the equatorial Pacific at 150 deg W

NASA Technical Reports Server (NTRS)

Stegmann, P. M.; Lewis, M. R.; Davis, C. O.; Cullen, J. J.

1992-01-01

Biological, optical, and hydrographical data were collected on the WEC88 cruise along 150 deg W and during a 6-day time-series station on the equator during February/March 1988. This area was characterized by a subsurface chlorophyll maximum (SCM), located at 50-70 m depth at the equator and descending down to 120-125 m at the north and south end of the transect. Highest primary production rates were near-surface and confined to the equatorial region and stations between 7 deg and 11 deg N. To determine the relationship between solar-stimulated fluorescence (centered at 683 nm wavelength) and primary production, a production-fluorescence model based on phytoplankton physiology and marine optics is described. Results of model calculations predict that there is a linear relation between production and fluorescence. A comparison between morning and midday measurements of the production-fluorescence relation showed that there was some difference between the two, whereas evening measurements, on the other hand, were distinctly different from the morning/midday ones. This seems to suggest that diurnal variations contribute significantly to variability in the quantum yield of photochemical processes. The ratio of the quantum yield of photosynthesis to the quantum yield of fluorescence ranged between 0.24 and 0.44 molC/Ein for all stations. The highest value for this ratio occurred at the equatorial stations, indicating that latitudinal variability could have an effect on the production-fluorescence relation.

Arthritis imaging using a near-infrared fluorescence folate-targeted probe

PubMed Central

Chen, Wei-Tsung; Mahmood, Umar; Weissleder, Ralph; Tung, Ching-Hsuan

2005-01-01

A recently developed near-infrared fluorescence-labeled folate probe (NIR2-folate) was tested for in vivo imaging of arthritis using a lipopolysaccharide intra-articular injection model and a KRN transgenic mice serum induction mouse model. In the lipopolysaccharide injection model, the fluorescence signal intensity of NIR2-folate (n = 12) and of free NIR2 (n = 5) was compared between lipopolysaccharide-treated and control joints. The fluorescence signal intensity of the NIR2-folate probe at the inflammatory joints was found to be significantly higher than the control normal joints (up to 2.3-fold, P < 0.001). The NIR2-free dye injection group showed a persistent lower enhancement ratio than the NIR2-folate probe injection group. Excessive folic acid was also given to demonstrate a competitive effect with the NIR2-folate. In the KRN serum transfer model (n = 4), NIR2-folate was applied at different time points after serum transfer, and the inflamed joints could be detected as early as 30 hours after arthritogenic antibody transfer (1.8-fold increase in signal intensity). Fluorescence microscopy, histology, and immunohistochemistry validated the optical imaging results. We conclude that in vivo arthritis detection was feasible using a folate-targeted near-infrared fluorescence probe. This receptor-targeted imaging method may facilitate improved arthritis diagnosis and early assessment of the disease progress by providing an in vivo characterization of active macrophage status in inflammatory joint diseases. PMID:15743478

Protoporphyrin IX fluorescence as potential indicator of psoriasis severity and progression.

PubMed

Wang, Bo; Xu, Yu-Ting; Zhang, Li; Zheng, Jie; Sroka, Ronald; Wang, Hong-Wei; Wang, Xiu-Li

2017-09-01

In psoriatic lesions, fluorescence diagnosis with blue light can detect protoporphyrin IX accumulation, especially after topical 5-aminolaevulinic acid (ALA) application. However, variable fluorescence distributions, interpersonal variations and long incubation time limit its wide application in clinic. This study is aimed to identify a consistent and convenient method to facilitate diagnosis and evaluation of psoriatic lesions. 104 psoriatic lesions from 30 patients were evaluated. Single lesion PSI scoring and fluorescence by macrospectrofluorometry were recorded on each lesion before and after treatment with narrow-band UVB. Punctate red fluorescence, emitted mainly by protoporphyrin IX, is observed in some psoriatic lesions. According to psoriasis severity index, fluorescence-positive lesions are more severe than lesions without fluorescence. We found a significant positive correlation between psoriasis severity and fluorescence intensity from protoporphyrin IX. Protoporphyrin IX-induced red fluorescence can be used as a novel and convenient approach for psoriasis diagnosis and progression evaluation. Copyright © 2017. Published by Elsevier B.V.

Test Program for Evaluation of Variable Frequency Power Conditioners

DOT National Transportation Integrated Search

1973-08-01

A test program is outlined for variable frequency power conditioners for 3-phase induction motors in vehicle propulsion applications. The Power Conditioner Unit (PCU) performance characteristics are discussed in some detail. Measurement methods, reco...

Weighted partial least squares based on the error and variance of the recovery rate in calibration set.

PubMed

Yu, Shaohui; Xiao, Xue; Ding, Hong; Xu, Ge; Li, Haixia; Liu, Jing

2017-08-05

The quantitative analysis is very difficult for the emission-excitation fluorescence spectroscopy of multi-component mixtures whose fluorescence peaks are serious overlapping. As an effective method for the quantitative analysis, partial least squares can extract the latent variables from both the independent variables and the dependent variables, so it can model for multiple correlations between variables. However, there are some factors that usually affect the prediction results of partial least squares, such as the noise, the distribution and amount of the samples in calibration set etc. This work focuses on the problems in the calibration set that are mentioned above. Firstly, the outliers in the calibration set are removed by leave-one-out cross-validation. Then, according to two different prediction requirements, the EWPLS method and the VWPLS method are proposed. The independent variables and dependent variables are weighted in the EWPLS method by the maximum error of the recovery rate and weighted in the VWPLS method by the maximum variance of the recovery rate. Three organic matters with serious overlapping excitation-emission fluorescence spectroscopy are selected for the experiments. The step adjustment parameter, the iteration number and the sample amount in the calibration set are discussed. The results show the EWPLS method and the VWPLS method are superior to the PLS method especially for the case of small samples in the calibration set. Copyright © 2017 Elsevier B.V. All rights reserved.

Synthesis, quantitative structure-property relationship study of novel fluorescence active 2-pyrazolines and application.

PubMed

Girgis, Adel S; Basta, Altaf H; El-Saied, Houssni; Mohamed, Mohamed A; Bedair, Ahmad H; Salim, Ahmad S

2018-03-01

A variety of fluorescence-active fluorinated pyrazolines 13-33 was synthesized in good yields through cyclocondensation reaction of propenones 1-9 with aryl hydrazines 10-12 . Some of the synthesized compounds provided promising fluorescence properties with quantum yield ( Φ ) higher than that of quinine sulfate (standard reference). Quantitative structure-property relationship studies were undertaken supporting the exhibited fluorescence properties and estimating the parameters governing properties. Five synthesized fluorescence-active pyrazolines ( 13 , 15 , 18 , 19 and 23 ) with variable Φ were selected for treating two types of paper sheets (Fabriano and Bible paper). These investigated fluorescence compounds, especially compounds 19 and 23 , provide improvements in strength properties of paper sheets. Based on the observed performance they can be used as markers in security documents.

Synthesis, quantitative structure–property relationship study of novel fluorescence active 2-pyrazolines and application

PubMed Central

Girgis, Adel S.; El-Saied, Houssni; Mohamed, Mohamed A.; Bedair, Ahmad H.; Salim, Ahmad S.

2018-01-01

A variety of fluorescence-active fluorinated pyrazolines 13–33 was synthesized in good yields through cyclocondensation reaction of propenones 1–9 with aryl hydrazines 10–12. Some of the synthesized compounds provided promising fluorescence properties with quantum yield (Φ) higher than that of quinine sulfate (standard reference). Quantitative structure–property relationship studies were undertaken supporting the exhibited fluorescence properties and estimating the parameters governing properties. Five synthesized fluorescence-active pyrazolines (13, 15, 18, 19 and 23) with variable Φ were selected for treating two types of paper sheets (Fabriano and Bible paper). These investigated fluorescence compounds, especially compounds 19 and 23, provide improvements in strength properties of paper sheets. Based on the observed performance they can be used as markers in security documents. PMID:29657796

Synthesis, quantitative structure-property relationship study of novel fluorescence active 2-pyrazolines and application

NASA Astrophysics Data System (ADS)

Girgis, Adel S.; Basta, Altaf H.; El-Saied, Houssni; Mohamed, Mohamed A.; Bedair, Ahmad H.; Salim, Ahmad S.

2018-03-01

A variety of fluorescence-active fluorinated pyrazolines 13-33 was synthesized in good yields through cyclocondensation reaction of propenones 1-9 with aryl hydrazines 10-12. Some of the synthesized compounds provided promising fluorescence properties with quantum yield (Φ) higher than that of quinine sulfate (standard reference). Quantitative structure-property relationship studies were undertaken supporting the exhibited fluorescence properties and estimating the parameters governing properties. Five synthesized fluorescence-active pyrazolines (13, 15, 18, 19 and 23) with variable Φ were selected for treating two types of paper sheets (Fabriano and Bible paper). These investigated fluorescence compounds, especially compounds 19 and 23, provide improvements in strength properties of paper sheets. Based on the observed performance they can be used as markers in security documents.

Ectopic bone formation cannot occur by hydroxyapatite/β-tricalcium phosphate bioceramics in green fluorescent protein chimeric mice

NASA Astrophysics Data System (ADS)

Cheng, Lijia; Duan, Xin; Xiang, Zhou; Shi, Yujun; Lu, Xiaofeng; Ye, Feng; Bu, Hong

2012-12-01

Many studies have shown that calcium phosphate ceramics (CP) have osteoconductive and osteoinductive properties; however, the exact mechanism of bone induction has not yet been reported. This study was performed to investigate if destroying immunological function will influence osteogenesis, to explain the mechanism which is unclear. In this study, twenty C57BL/6 mice were divided into two groups (n = 10), in group 1, a hydroxyapatite/β-tricalcium phosphate (HA/β-TCP) ceramic was implanted into both the left and right leg muscles of each mouse; in group 2, ten mice experienced lethal irradiation, then were injected bone marrow (BM) cells from green fluorescent protein (GFP) transgenic mice by tail veil, after bone marrow transplantation (BMT), heart, liver, spleen, lung, kidney, and muscle were harvested for biological analysis, after the GFP chimera model was established successfully, the same HA/β-TCP ceramic was implanted into both leg muscles of each mouse immediately after irradiation. 45 and 90 days after implantation, the ceramics of the two groups were harvested to perform with hematoxylin and eosin (HE) and immunohistochemistry (IHC) staining; the results showed that there was no bone formation in group 2, while new bone tissues were detected in group 1. Our findings suggest that the BM cell from GFP transgenic mice is a good biomarker and it could set a good platform for chimera model; it also shows that BM cell is one of cell resources of bone induction, and destruction of immune function will impede osteoinduction by CP. Overall, our results may shed light on clear mechanism study of bone induction in the future.

Sensitivity evaluation of the green alga Chlamydomonas reinhardtii to uranium by pulse amplitude modulated (PAM) fluorometry.

PubMed

Herlory, Olivier; Bonzom, Jean-Marc; Gilbin, Rodolphe

2013-09-15

Although ecotoxicological studies tend to address the toxicity thresholds of uranium in freshwaters, there is a lack of information on the effects of the metal on physiological processes, particularly in aquatic plants. Knowing that uranium alters photosynthesis via impairment of the water photo-oxidation process, we determined whether pulse amplitude modulated (PAM) fluorometry was a relevant tool for assessing the impact of uranium on the green alga Chlamydomonas reinhardtii and investigated how and to what extent uranium hampered photosynthetic performance. Photosynthetic activity and quenching were assessed from fluorescence induction curves generated by PAM fluorometry, after 1 and 5h of uranium exposure in controlled conditions. The oxygen-evolving complex (OEC) of PSII was identified as the primary action site of uranium, through alteration of the water photo-oxidation process as revealed by F0/Fv. Limiting re-oxidation of the plastoquinone pool, uranium impaired the electron flux between the photosystems until almost complete inhibition of the PSII quantum efficiency ( [Formula: see text] , EC50=303 ± 64 μg UL(-1) after 5h of exposure) was observed. Non-photochemical quenching (qN) was identified as the most sensitive fluorescence parameter (EC50=142 ± 98 μg UL(-1) after 5h of exposure), indicating that light energy not used in photochemistry was dissipated in non-radiative processes. It was shown that parameters which stemmed from fluorescence induction kinetics are valuable indicators for evaluating the impact of uranium on PSII in green algae. PAM fluorometry provided a rapid and reasonably sensitive method for assessing stress response to uranium in microalgae. Copyright © 2013 Elsevier B.V. All rights reserved.

Curcumin induced autophagy anticancer effects on human lung adenocarcinoma cell line A549

PubMed Central

Liu, Furong; Gao, Song; Yang, Yuxuan; Zhao, Xiaodan; Fan, Yameng; Ma, Wenxia; Yang, Danrong; Yang, Aimin; Yu, Yan

2017-01-01

To investigate the anticancer effects of curcumin-induced autophagy and its effects on the human lung adenocarcinoma A549 cell line, inverted phase contrast microscopy was used to observe alterations to the cytomorphology of cells. An MTT assay was used to measure cell viability. Autophagy was detected using acridine orange (AO) staining and 3-methyladenine (3-MA) was used as an autophagy-specific inhibitor. Dose- and time-dependent A549 cell viability inhibition was observed following curcumin treatment. A dose-dependent increase in the red fluorescent structures in A549 cells was identified following curcumin treatment for 48 h through AO staining. In addition, the activation of autophagy was determined through changes in the number of autophagic vesicles (AVs; fluorescent particles) infected with monodansylcadaverine (MDC). The fluorescence intensity and density of AVs in the curcumin-treated groups were higher at 48 h compared with the control group. Finally, the MTT assay demonstrated that the survival rates of the curcumin-treated cells were increased when pretreated with 3-MA for 3 h, indicating that the inhibitory effect of curcumin on A549 cells is reduced following the inhibition of autophagy. Furthermore, AO and MDC staining confirmed that 3-MA does inhibit the induction of autophagy. Thus, it was hypothesized that the induction of autophagy is partially involved in the reduction of cell viability observed following curcumin treatment. The anticancer effects of curcumin on A549 cells can be reduced using autophagy inhibitors. This suggests a possible cancer therapeutic application of curcumin through the activation of autophagy. These findings have improved the understanding of the mechanism underlying the anticancer property of curcumin. PMID:28928819

A high-throughput quantitative expression analysis of cancer-related genes in human HepG2 cells in response to limonene, a potential anticancer agent.

PubMed

Hafidh, Rand R; Hussein, Saba Z; MalAllah, Mohammed Q; Abdulamir, Ahmed S; Abu Bakar, Fatimah

2017-11-14

Citrus bioactive compounds, as active anticancer agent, have been under focus by several studies worldwide. However, the underlying genes responsible for the anticancer potential have not been sufficiently highlighted. The current study investigated the gene expression profile of hepatocellular carcinoma, HepG2, cells after treatment with Limonene. The concentration that killed 50% of HepG2 cells was used to elucidate the genetic mechanisms of limonene anticancer activity. The apoptotic induction was detected by flow cytometry and confocal fluorescence microscope. Two of pro-apoptotic events, caspase-3 activation and phosphatidylserine translocation were manifested by confocal fluorescence microscopy. High-throughput real-time PCR was used to profile 1023 cancer-related genes in 16 different gene families related to the cancer development. In comparison to untreated cells, limonene increased the percentage of apoptotic cells up to 89.61%, by flow cytometry, and 48.2% by fluorescence microscopy. There was a significant limonene-driven differential gene expression of HepG2 cells in 15 different gene families. Limonene was shown to significantly (>2log) up-regulate and down-regulate 14 and 59 genes, respectively. The affected gene families, from most to least affected, were apoptosis induction, signal transduction, cancer genes augmentation, alteration in kinases expression, inflammation, DNA damage repair, and cell cycle proteins. The current study reveals that limonene could be a promising, cheap, and effective anticancer compound. The broad spectrum of limonene anticancer activity is interesting for anticancer drug development. Further research is needed to confirm the current findings and to examine the anticancer potential of limonene along with underlying mechanisms on different cell lines. Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.

Curcumin induced autophagy anticancer effects on human lung adenocarcinoma cell line A549.

PubMed

Liu, Furong; Gao, Song; Yang, Yuxuan; Zhao, Xiaodan; Fan, Yameng; Ma, Wenxia; Yang, Danrong; Yang, Aimin; Yu, Yan

2017-09-01

To investigate the anticancer effects of curcumin-induced autophagy and its effects on the human lung adenocarcinoma A549 cell line, inverted phase contrast microscopy was used to observe alterations to the cytomorphology of cells. An MTT assay was used to measure cell viability. Autophagy was detected using acridine orange (AO) staining and 3-methyladenine (3-MA) was used as an autophagy-specific inhibitor. Dose- and time-dependent A549 cell viability inhibition was observed following curcumin treatment. A dose-dependent increase in the red fluorescent structures in A549 cells was identified following curcumin treatment for 48 h through AO staining. In addition, the activation of autophagy was determined through changes in the number of autophagic vesicles (AVs; fluorescent particles) infected with monodansylcadaverine (MDC). The fluorescence intensity and density of AVs in the curcumin-treated groups were higher at 48 h compared with the control group. Finally, the MTT assay demonstrated that the survival rates of the curcumin-treated cells were increased when pretreated with 3-MA for 3 h, indicating that the inhibitory effect of curcumin on A549 cells is reduced following the inhibition of autophagy. Furthermore, AO and MDC staining confirmed that 3-MA does inhibit the induction of autophagy. Thus, it was hypothesized that the induction of autophagy is partially involved in the reduction of cell viability observed following curcumin treatment. The anticancer effects of curcumin on A549 cells can be reduced using autophagy inhibitors. This suggests a possible cancer therapeutic application of curcumin through the activation of autophagy. These findings have improved the understanding of the mechanism underlying the anticancer property of curcumin.

Processes Affecting the Variability of Fluorescence Signals from Benthic Targets in Shallow Waters

DTIC Science & Technology

1999-09-30

isolated green fluorescent proteins from a variety of cnidarians and characterized their spectral signatures and biochemical characteristics. RESULTS...types of GFP-like fluorophores are present in Cnidarians . The first one had the only excitation band centered at ca. 495 nm; the second type had two

The influence of low-frequency magnetic field on plasma antioxidant capacity and heart rate.

PubMed

Ciejka, Elzbieta B; Goraca, Anna

2009-01-01

Low-frequency magnetic field is widely applied as magnetotherapy in physiotherapeutic treatment. Recognition of positive and negative effects of the magnetic field has been the subject of numerous studies. Experimental studies concern, among others, the effect of this field on the heart rate and plasma antioxidant capacity. The aim of the study was to check whether a time-variable magnetic field of constant frequency and induction affects the heart rate and plasma antioxidant capacity. The tests were performed on Spraque-Dawley rats exposed to the magnetic field of the following parameters: frequency - 40 Hz, induction - 7 mT, time of exposure - 30 and 60 minutes. The measurements of ECG and plasma antioxidant capacity expressed in the number of reduced iron ions were performed on experimental animals: before, after a single exposure and after 14 days of exposure. A significant decrease of the heart rate was observed after 14 days of exposure. A variable magnetic field of the parameters: frequency - 40 Hz, induction - 7 mT and exposure time of 14 days caused an increase of the organism antioxidant defence, whereas a variable magnetic field of the frequency of 40 Hz, induction - 7 mT and exposure time 60 minutes for 14 days caused a significant decrease of the organism antioxidant defence. The exposure time affects heart rate, plasma antioxidant capacity and the organism defense ability against free radicals.

An interlaboratory comparison of bone lead measurements via K-shell X-ray fluorescence spectrometry: validation against inductively coupled plasma mass spectrometry

PubMed Central

Bellis, David J.; Todd, Andrew C.

2012-01-01

109Cd-based K-shell X-ray fluorescence spectrometry (hereafter, for brevity, XRF) is used, often in epidemiological studies, to perform non-invasive, in vivo measurements of lead in bone. We conducted the first interlaboratory study of XRF via the circulation of nine goat tibiæ in which the mean lead value ranged from 4.0 µg g−1 to 55.3 µg g−1 bone mineral. The test tibiæ were subsequently analyzed via nitric acid digestion followed by lead determination by inductively coupled plasma mass spectrometry (ICP-MS) – along with certified reference materials for bone lead – thus providing measurement traceability to SI units. Analysis of dried bone for lead via nitric acid digestion and ICP-MS yields mass fraction data in units of µg g−1 dry weight. The mean bone lead value based on ICP-MS analysis ranged from 1.8 µg g−1 to 35.8 µg g−1 dry weight. For comparison purposes, XRF-measured Pb values (µg g−1 bone mineral) were converted into the ICP-MS-measured units (µg g−1dry weight bone) by multiplying the former by the average ash fraction from the nine tibiæ. Eight of the XRF systems did not yield a significant bias for any of the nine tibiæ; one system was biased for one of the tibiæ; two systems were biased for two tibiæ; one system was biased for four tibiæ; two systems (813-1 and 804-2) were biased for five tibiæ and one system (801-1) was biased for six of the nine tibiæ. Average bias for the systems (under those particular operating conditions) that were biased for the majority of samples ranged from −2.6 µg g−1 (−15.7%) to 5.1 µg g−1 (30.7%) dry weight bone. All participants now have the ICP-MS data, allowing any corrective actions deemed necessary to be implemented. The ICP-MS data, however, indicated that the lead mass fraction varied considerably with the sampling location within the tibiæ, to the extent of exceeding XRF variability for the higher lead values. Material heterogeneity is an unavoidable reality of measuring lead in bone. PMID:22468015

Induction powered biological radiosonde

NASA Technical Reports Server (NTRS)

Fryer, T. B. (Inventor)

1980-01-01

An induction powered implanted monitor for epidurally measuring intracranial pressure and telemetering the pressure information to a remote readout is disclosed. The monitor utilizes an inductance-capacitance (L-C) oscillator in which the C comprises a variable capacitance transducer, one electrode of which is a small stiff pressure responsive diaphragm. The oscillator is isolated from a transmitting tank circuit by a buffer circuit and all electric components in the implanted unit except an input and an output coil are shielded by a metal housing.

Heterogeneous distribution of antigens on human platelets demonstrated by fluorescence flow cytometry.

PubMed

Dunstan, R A; Simpson, M B

1985-12-01

We have used fluorescence flow cytometry to analyse cell-to-cell variability in the density of platelet ABH, Ii, Lewis, P, P1A1, Bak,a and HLA class I antigens. Human IgG and IgM antibodies were used in a two-stage assay with goat FITC-conjugated antihuman IgG (H&L) antibody as the label, followed by single cell analysis of 10 000 platelets per sample using a 256-channel fluorescence flow cytometer (Becton-Dickinson FACS Analyser). Computer analysis of fluorescence intensity histograms for mean and peak channel and coefficient of variation shows that the degree of heterogeneity in platelet antigen density varies with each particular blood group. The broad fluorescence distribution curves with oligosaccharide antigens (CVs: A = 53, B = 40, I = 44, Lea = 40, P = 40) indicate that these antigens possess a greater variability in the number of sites per cell compared to the more homogeneous distribution of P1,A1 BaK,a and HLA (CVs: P1A1 = 24, HLA = 30). These findings may partly account for the mechanism by which transfusion of ABO-incompatible platelets results in a biphasic survival curve, with a period of early rapid removal of those platelets with a high density of antigen sites, followed by a relatively normal survival curve for those platelets that possess only a few or no antigen sites. In contrast, P1A1 and HLA sites are less variable in number from one platelet to another in a given donor, and immune-mediated removal would be more likely to approximate a single exponential curve.

Introduction to the special section ‘Applications of electromagnetic induction to digital soil mapping’

USDA-ARS?s Scientific Manuscript database

Use of electromagnetic induction (EMI) instruments has increased as a tool to map soils because it provides a means of locating suitable sampling sites that provide the basis for mapping the spatial variability of various soil properties either directly or indirectly measured with EMI, including sa...

Arrhenius Rate: constant volume burn

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

Menikoff, Ralph

A constant volume burn occurs for an idealized initial state in which a large volume of reactants at rest is suddenly raised to a high temperature and begins to burn. Due to the uniform spatial state, there is no fluid motion and no heat conduction. This reduces the time evolu tion to an ODE for the reaction progress variable. With an Arrhenius reaction rate, two characteristics of thermal ignition are illustrated: induction time and thermal runaway. The Frank-Kamenetskii approximation then leads to a simple expression for the adiabatic induction time. For a first order reaction, the analytic solution is derivedmore » and used to illustrate the effect of varying the activation temperature; in particular, on the induction time. In general, the ODE can be solved numerically. This is used to illustrate the effect of varying the reaction order. We note that for a first order reaction, the time evolution of the reaction progress variable has an exponential tail. In contrast, for a reaction order less than one, the reaction completes in a nite time. The reaction order also affects the induction time.« less

Spectral and Temporal Laser Fluorescence Analysis Such as for Natural Aquatic Environments

NASA Technical Reports Server (NTRS)

Chekalyuk, Alexander (Inventor)

2015-01-01

An Advanced Laser Fluorometer (ALF) can combine spectrally and temporally resolved measurements of laser-stimulated emission (LSE) for characterization of dissolved and particulate matter, including fluorescence constituents, in liquids. Spectral deconvolution (SDC) analysis of LSE spectral measurements can accurately retrieve information about individual fluorescent bands, such as can be attributed to chlorophyll-a (Chl-a), phycobiliprotein (PBP) pigments, or chromophoric dissolved organic matter (CDOM), among others. Improved physiological assessments of photosynthesizing organisms can use SDC analysis and temporal LSE measurements to assess variable fluorescence corrected for SDC-retrieved background fluorescence. Fluorescence assessments of Chl-a concentration based on LSE spectral measurements can be improved using photo-physiological information from temporal measurements. Quantitative assessments of PBP pigments, CDOM, and other fluorescent constituents, as well as basic structural characterizations of photosynthesizing populations, can be performed using SDC analysis of LSE spectral measurements.

[Construction of injectable tissue engineered adipose tissue with fibrin glue scaffold and human adipose-derived stem cells transfected by lentivirus vector expressing hepatocyte growth factor].

PubMed

Zhu, Yuanzheng; Yi, Yangyan; Yang, Shuifa; Zhang, Jing; Wu, Shu; Wang, Zhaohui

2017-09-01

To discuss the possibility of constructing injectable tissue engineered adipose tissue, and to provide a new approach for repairing soft tissue defects. Human adipose-derived stem cells (hADSCs) were extracted from the lipid part of human liposuction aspirate by enzymatic digestion and identified by morphological observation, flow cytometry, and adipogenic induction. The hADSCs underwent transfection by lentivirus vector expressing hepatocyte growth factor and green fluorescent protein (HGF-GFP-LVs) of different multiplicity of infection (MOI, 10, 30, 50, and 100), the transfection efficiency was calculated to determine the optimum MOI. The hADSCs transfected by HGF-GFP-LVs of optimal MOI and being adipogenic inducted were combined with injectable fibrin glue scaffold, and were injected subcutaneously into the right side of the low back of 10 T-cell deficiency BALB/c female nude mice (transfected group); non-HGF-GFP-LVs transfected hADSCs (being adipogenic inducted) combined with injectable fibrin glue scaffold were injected subcutaneously into the left side of the low back (untransfected group); and injectable fibrin glue scaffold were injected subcutaneously into the middle part of the neck (blank control group); 0.4 mL at each point. Twelve weeks later the mice were killed and the implants were taken out. Gross observation, wet weight measurement, HE staining, GFP fluorescence labeling, and immunofluorescence staining were performed to assess the in vivo adipogenic ability of the seed cells and the neovascularization of the grafts. The cultured cells were identified as hADSCs. Poor transfection efficiency was observed in MOI of 10 and 30, the transfection efficiency of MOI of 50 and 100 was more than 80%, so the optimum MOI was 50. Adipose tissue-like new-born tissues were found in the injection sites of the transfected and untransfected groups after 12 weeks of injection, and no new-born tissues was found in the blank control group. The wet-weight of new-born tissue in the transfected group [(32.30±4.06) mg] was significantly heavier than that of the untransfected group [(25.27±3.94) mg] ( t =3.929, P =0.001). The mature adipose cells in the transfected group [(126.93±5.36) cells/field] were significantly more than that in the untransfected group [(71.36±4.52) cells/field] ( t =30.700, P =0.000). Under fluorescence microscopy, some of the single cell adipocytes showed a network of green fluorescence, indicating the presence of GFP labeled exogenous hADSCs in the tissue. The vascular density of new-born tissue of the transfected group [(16.37±2.76)/field] was significantly higher than that of the untransfected group [(9.13±1.68)/field] ( t =8.678, P =0.000). The hADSCs extracted from the lipid part after liposuction can be used as seed cells. After HGF-GFP-LVs transfection and adipose induction, the hADSCs combined with injectable fibrin glue scaffold can construct mature adipose tissue in vivo , which may stimulate angiogenesis, and improve retention rate of new-born tissue.

Physical and geochemical drivers of CDOM variability near a natural seep site in the Gulf of Mexico

NASA Astrophysics Data System (ADS)

Edwards, C. R.; Powers, L.; Medeiros, P. M.

2016-02-01

Colored dissolved organic matter (CDOM) on the continental shelf and slope can serve as a marker for fresh water influence, indicate the presence of hydrocarbons, and provide important clues about nutrient content and organic matter cycling. Autonomous underwater vehicles such as gliders allow for subsurface measurement of CDOM fluorescence for weeks to months; these time series may be especially valuable in the northern Gulf of Mexico, where CDOM inputs of both terrestrial and oil and gas sources can be significant. Data from a recent glider deployment near a natural seep site (GC600) on the continental slope over 180km from shore suggest simultaneous influence of Mississippi plume water and hydrocarbon inputs in the upper 200m, with variability in fluorescence at a range of vertical and temporal scales. We will explore patterns in spatial and temporal variability of glider-measured hydrography, dissolved oxygen, and bio-optical data (CDOM, chlorophyll-a, backscatter fluorescence), and use their combination to infer a terrigenous and/or fossil fuel source(s). Taking advantage of a combination of satellite sea surface temperature, ocean color, wind, and data from moored and mobile platforms, we will examine physical controls on transport and vertical mixing of CDOM and the potential role of nonlinear mesoscale eddies, which can trap water in their interior and may transport river- or hydrocarbon-derived CDOM over long distances. The combined data set will be used to consider and potentially constrain the effect of photodegradation and other biogeochemical causes for CDOM fluorescence variability in the upper 200m.

Optical fluorescence biosensor for plant water stress detection

NASA Astrophysics Data System (ADS)

Chong, Jenny P. C.; Liew, O. W.; Li, B. Q.; Asundi, A. K.

2007-05-01

Precision farming in arable agriculture and horticulture allows conservative use of resources that are applied according to plant needs. The growing concern for sustainability in crop production has accentuated the significance of our work to develop a rapid, sensitive and non-destructive spectroscopic method for real-time monitoring of plant water stress. Elucidation of crop water status before the onset of irreversible cellular damage is critical for effective water management to ensure maximum crop yield and profit margin. A two-component bio-sensing system comprising transgenic 'Indicator Plants' and a spectrometer-linked stereoscopic microscope was developed to detect early signs of water stress before the permanent wilting point is reached. The 'Indicator Plants' are transgenic Petunia hybrida genetically engineered with a drought-responsive promoter-linked enhanced green fluorescent protein marker gene (EGFP). No EGFP fluorescence was detected prior to induction of dehydration stress. Fluorescence emission intensity increased with dehydration period and was found mainly in the stems, leaf veins and leaf tips. While fluorescence emission above endogenous background was detectable after 2 hours of water stress treatment, the plants reached permanent wilting point after 6 hours, showing that our system was able to detect water stress prior to plant entry into the stage of irreversible damage. Future work will be geared towards overcoming biological and instrument-related difficulties encountered in our initial detection system.

HS-133, a novel fluorescent phosphatidylinositol 3-kinase inhibitor as a potential imaging and anticancer agent for targeted therapy

PubMed Central

Lee, Hyunseung; Son, Mi Kwon; Yun, Sun-Mi; Ahn, Sung-Hoon; Lee, Kyeong-Ryoon; Lee, Soyoung; Kim, Donghee; Hong, Sungwoo; Hong, Soon-Sun

2014-01-01

As PI3K/Akt signaling is frequently deregulated in a wide variety of human tumors, PI3K inhibitors are an emerging class of drugs for cancer treatment. The monitoring of the drug behavior and distribution in the biological system can play an important role for targeted therapy and provide information regarding the response or resistance to available therapies. In this study, therefore, we have developed a family of xanthine derivatives, serving as a dual function exhibiting fluorescence, as well as inhibiting PI3K. Among them, HS-133 showed anti-proliferative effects and was monitored for its subcellular localization by a fluorescence microscopy. HS-133 suppressed the PI3K/Akt pathway and induced cell cycle arrest at the G0/G1 phase. The induction of apoptosis by HS-133 was confirmed by the increases of the cleaved PARP, caspase-3, and caspase-8. Furthermore, HS-133 decreased the protein expression of HIF-1α and VEGF, as well inhibited the tube formation and migration of the human umbilical vein endothelial cells. In vivo imaging also showed that tumors were visualized fluorescent with HS-133, and its oral administration significantly inhibited the growth of tumor in SkBr3 mouse xenograft models. Thus, we suggest that HS-133 may be used as a fluorescent anticancer agent against human breast cancer. PMID:25338206

HS-133, a novel fluorescent phosphatidylinositol 3-kinase inhibitor as a potential imaging and anticancer agent for targeted therapy.

PubMed

Lee, Ju-Hee; Jung, Kyung Hee; Lee, Hyunseung; Son, Mi Kwon; Yun, Sun-Mi; Ahn, Sung-Hoon; Lee, Kyeong-Ryoon; Lee, Soyoung; Kim, Donghee; Hong, Sungwoo; Hong, Soon-Sun

2014-10-30

As PI3K/Akt signaling is frequently deregulated in a wide variety of human tumors, PI3K inhibitors are an emerging class of drugs for cancer treatment. The monitoring of the drug behavior and distribution in the biological system can play an important role for targeted therapy and provide information regarding the response or resistance to available therapies. In this study, therefore, we have developed a family of xanthine derivatives, serving as a dual function exhibiting fluorescence, as well as inhibiting PI3K. Among them, HS-133 showed anti-proliferative effects and was monitored for its subcellular localization by a fluorescence microscopy. HS-133 suppressed the PI3K/Akt pathway and induced cell cycle arrest at the G0/G1 phase. The induction of apoptosis by HS-133 was confirmed by the increases of the cleaved PARP, caspase-3, and caspase-8. Furthermore, HS-133 decreased the protein expression of HIF-1α and VEGF, as well inhibited the tube formation and migration of the human umbilical vein endothelial cells. In vivo imaging also showed that tumors were visualized fluorescent with HS-133, and its oral administration significantly inhibited the growth of tumor in SkBr3 mouse xenograft models. Thus, we suggest that HS-133 may be used as a fluorescent anticancer agent against human breast cancer.

Early quantitative method for measuring germination in non-green spores of Dryopteris paleacea using an epifluorescence-microscope technique

NASA Technical Reports Server (NTRS)

Scheuerlein, R.; Wayne, R.; Roux, S. J.

1988-01-01

A method is described to determine germination by blue-light excited red fluorescence in the positively photoblastic spores of Dryopteris paleacea Sw. This fluorescence is due to chlorophyll as evidenced from 1) a fluorescence-emission spectrum in vivo, where a bright fluorescence around 675 nm is obtained only in red light (R)-irradiated spores and 2) in vitro measurements with acetone extracts prepared from homogenized spores. Significant amounts of chlorophyll can be found only in R-treated spores; this chlorophyll exhibits an emission band around 668 nm, when irradiated with 430 nm light at 21 degrees C. Compared to other criteria for germination, such as swelling of the cell, coat splitting, greening, and rhizoid formation, which require longer periods after induction for their expression, chlorophyll fluorescence can be used to quantify germination after two days. This result is confirmed by fluence-response curves for R-induced spore germination; the same relationship between applied R and germination is obtained by the evaluation with the epifluorescence method 2 days after the light treatment as compared with the evaluation with bright-field microscopy 5 days after the inducing R. Using this technique we show for the first time that Ca2+ contributes to the signal-transduction chain in phytochrome-mediated chlorophyll synthesis in spores of Dryopteris paleacea.

Peroxynitrite-induced structural perturbations in human IgG: A physicochemical study.

PubMed

Arfat, Mir Yasir; Arif, Zarina; Chaturvedi, Sumit Kumar; Moinuddin; Alam, Khursheed

2016-08-01

IgG is an important defence protein. To exhibit optimum function the molecule must maintain its native structure. Peroxynitrite is a potent oxidizing and nitrating agent produced in vivo under pathophysiological conditions. It can oxidize and/or nitrate various amino acids causing changes in the structure and function of proteins. Such proteins may be involved in the pathogenesis of many inflammatory diseases, including rheumatoid arthritis. In the present work, peroxynitrite-induced structural changes in IgG have been studied by UV-visible, fluorescence, CD, FT-IR, DLS spectroscopy and DSC as well as by SDS-PAGE. Peroxynitrite-modified IgG exhibited hyperchromicity at 280 nm, quenching of tryptophan fluorescence, increase in ANS fluorescence, loss of β-sheet, shift in the positions of amide I and amide II bands, appearance of new peak in FT-IR, attachment of nitro residues and increase in melting temperature, compared to native IgG. Furthermore, peroxynitrite-modified IgG exhibited an additional peak at 420 nm, quenching in tyrosine fluorescence and enhancement in dityrosine fluorescence compared to native IgG. Generation of nitrotyrosine, dityrosine and nitrotryptophan was also observed in peroxynitrite-modified IgG. Gross structural changes in IgG caused by peroxynitrite and observed in vitro may favour autoantibodies induction in vivo under similar conditions. Copyright © 2016 Elsevier Inc. All rights reserved.

A theranostic nanoplatform: magneto-gold@fluorescence polymer nanoparticles for tumor targeting T1&T2-MRI/CT/NIR fluorescence imaging and induction of genuine autophagy mediated chemotherapy.

PubMed

Wang, Guannan; Qian, Kun; Mei, Xifan

2018-06-14

Multifunctional nanoparticles, bearing low toxicity and tumor-targeting properties, coupled with multifunctional diagnostic imaging and enhanced treatment efficacy, have drawn tremendous attention due to their enormous potential for medical applications. Herein, we report a new kind of biocompatible and tumor-targeting magneto-gold@fluorescent polymer nanoparticle (MGFs-LyP-1), which is based on ultra-small magneto-gold (Fe 3 O 4 -Au) nanoparticles and NIR emissive fluorescent polymers by a solvent-mediated method. This kind of nanoparticle could be taken up efficiently and simultaneously serve for in vivo tumor targeting T 1 &T 2 -MRI/CT/near infrared (NIR) fluorescence bioimaging. Furthermore, the nanoparticles exhibit small size, higher tumor targeting accumulation, excellent cytocompatibility for long-term tracking, and no disturbing cell proliferation and differentiation. Moreover, clear and convincing evidence proves that as-synthesized MGFs-LyP-1 could elicit genuine autophagy via inducing autophagosome formation, which offers a definite synergistic effect to enhance cancer therapy with doxorubicin (DOX) at a nontoxic concentration through enhancement of the autophagy flux. Meanwhile, the as-prepared nanoparticles could be rapidly cleared from mice without any obvious organ impairment. The results indeed reveal a promising prospect of an MGFs-LyP-1 contrast agent with low toxicity and high efficiency for promising application in biomedicine.

A variable-speed, constant-frequency wind power generation scheme using a slip-ring induction generator

NASA Astrophysics Data System (ADS)

Velayudhan, C.; Bundell, J. H.

This paper investigates a variable-speed, constant-frequency double output induction generator which is capable of absorbing the mechanical energy from a fixed pitch wind turbine and converting it into electrical energy at constant grid voltage and frequency. Rotor power at varying voltage and frequency is either fed to electronically controlled resistances and used as heat energy or is rectified, inverted by a controllable line-commutated inverter and returned to the grid. Optimal power tracking is by means of an adaptive controller which controls the developed torque of the generator by monitoring the shaft speed.

[Experimental analysis of some determinants of inductive reasoning].

PubMed

Ono, K

1989-02-01

Three experiments were conducted from a behavioral perspective to investigate the determinants of inductive reasoning and to compare some methodological differences. The dependent variable used in these experiments was the threshold of confident response (TCR), which was defined as "the minimal sample size required to establish generalization from instances." Experiment 1 examined the effects of population size on inductive reasoning, and the results from 35 college students showed that the TCR varied in proportion to the logarithm of population size. In Experiment 2, 30 subjects showed distinct sensitivity to both prior probability and base-rate. The results from 70 subjects who participated in Experiment 3 showed that the TCR was affected by its consequences (risk condition), and especially, that humans were sensitive to a loss situation. These results demonstrate the sensitivity of humans to statistical variables in inductive reasoning. Furthermore, methodological comparison indicated that the experimentally observed values of TCR were close to, but not as precise as the optimal values predicted by Bayes' model. On the other hand, the subjective TCR estimated by subjects was highly discrepant from the observed TCR. These findings suggest that various aspects of inductive reasoning can be fruitfully investigated not only from subjective estimations such as probability likelihood but also from an objective behavioral perspective.

B cell epitopes on infliximab identified by oligopeptide microarray with unprocessed patient sera.

PubMed

Homann, Arne; Röckendorf, Niels; Kromminga, Arno; Frey, Andreas; Jappe, Uta

2015-10-29

Autoimmune diseases like rheumatoid arthritis and inflammatory bowel disease are treated with TNF-alpha-blocking antibodies such as infliximab and adalimumab. A common side effect of therapeutic antibodies is the induction of anti-drug antibodies, which may reduce therapeutic efficacy. In order to reveal immunogenic epitopes on infliximab which are responsible for the adverse effects, sera from patients treated with infliximab were screened by ELISA for anti-infliximab antibodies. Sera containing high levels of anti-drug-antibodies (>1.25 µg/ml) were analyzed in an oligopeptide microarray system containing immobilized 15-meric oligopeptides from the infliximab amino acid sequence. Immunogenic infliximab IgG-epitopes were identified by infrared fluorescence scanning and comparison of infliximab-treated patients versus untreated controls. Six relevant epitopes on infliximab were recognized by the majority of all patient sera: 4 in the variable and 2 in the constant region. Three of the epitopes in the variable region are located in the TNF-alpha binding region of infliximab. The fourth epitope of the variable part of infliximab is located close to the TNF-alpha binding region and contains an N-glycosylation sequon. The sera positive for anti-infliximab antibodies do not contain antibodies against adalimumab as determined by ELISA. Thus, there is no infliximab-adalimumab cross-reactivity as determined by these systems. Our data shall contribute to a knowledge-based recommendation for a potentially necessary therapy switch from infliximab to another type of TNF-alpha-blocker. The characterization of immunogenic epitopes on therapeutic monoclonal antibodies using unprocessed patient sera shall lead to direct translational aspects for the development of less immunogenic therapeutic antibodies. Patients benefit from less adverse events and longer lasting drug effects.

Iron triggers λSo prophage induction and release of extracellular DNA in Shewanella oneidensis MR-1 biofilms.

PubMed

Binnenkade, Lucas; Teichmann, Laura; Thormann, Kai M

2014-09-01

Prophages are ubiquitous elements within bacterial chromosomes and affect host physiology and ecology in multiple ways. We have previously demonstrated that phage-induced lysis is required for extracellular DNA (eDNA) release and normal biofilm formation in Shewanella oneidensis MR-1. Here, we investigated the regulatory mechanisms of prophage λSo spatiotemporal induction in biofilms. To this end, we used a functional fluorescence fusion to monitor λSo activation in various mutant backgrounds and in response to different physiological conditions. λSo induction occurred mainly in a subpopulation of filamentous cells in a strictly RecA-dependent manner, implicating oxidative stress-induced DNA damage as the major trigger. Accordingly, mutants affected in the oxidative stress response (ΔoxyR) or iron homeostasis (Δfur) displayed drastically increased levels of phage induction and abnormal biofilm formation, while planktonic cells were not or only marginally affected. To further investigate the role of oxidative stress, we performed a mutant screen and identified two independent amino acid substitutions in OxyR (T104N and L197P) that suppress induction of λSo by hydrogen peroxide (H2O2). However, λSo induction was not suppressed in biofilms formed by both mutants, suggesting a minor role of intracellular H2O2 in this process. In contrast, addition of iron to biofilms strongly enhanced λSo induction and eDNA release, while both processes were significantly suppressed at low iron levels, strongly indicating that iron is the limiting factor. We conclude that uptake of iron during biofilm formation triggers λSo-mediated lysis of a subpopulation of cells, likely by an increase in iron-mediated DNA damage sensed by RecA. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

Investigation of the electrical discharge parameters in electrodeless inductive lamps with a re-entrant coupler and magnetic core

NASA Astrophysics Data System (ADS)

Statnic, Eugen; Tanach, Valentin

2006-08-01

The inductively coupled fluorescent lamp with a cored induction coil placed in a re-entrant cavity is in fact a coaxial transformer operated in the radiofrequency range between 100 kHz and a few MHz. The magnetic coupling coefficient k between the primary coil and the plasma ring is relatively low because of the open magnetic circuit. The acting mutual inductance M enables us to quantify the interaction between the magnetic field produced by the primary coil current I1 and the opposing magnetic field produced by the powerful plasma current I2. A contra-electromotive force jωMI2 is induced in the induction coil L1, defining the primary voltage V1 = I1(R1 + jω L1) - jωMI2. The current I1 induces in the secondary conductive plasma the driving electromotive force jωMI1 supplying the secondary load consisting of the average plasma resistance R2 and the secondary inductance L2, according to the equation jωMI2 = I2(R2 + jωL2). It is the aim of this paper to find a model to determine k on the basis of the measured primary parameters V1, I1, P1, L1, R1 and finally all electrical inaccessible parameters, such as M, V2, I2, phiv2, L2, R2, in order to optimize the discharge and lamp efficacy. The complex characteristic of plasma inductance for this type of lamp is analysed and clarified. Some reflexive basic relations verifying the correctness of the inferred plasma parameters are also developed. The described experiments are related to a lamp working at about 2.6 MHz.

Analysis of SOS-Induced Spontaneous Prophage Induction in Corynebacterium glutamicum at the Single-Cell Level

PubMed Central

Nanda, Arun M.; Heyer, Antonia; Krämer, Christina; Grünberger, Alexander; Kohlheyer, Dietrich

2014-01-01

The genome of the Gram-positive soil bacterium Corynebacterium glutamicum ATCC 13032 contains three integrated prophage elements (CGP1 to -3). Recently, it was shown that the large lysogenic prophage CGP3 (∼187 kbp) is excised spontaneously in a small number of cells. In this study, we provide evidence that a spontaneously induced SOS response is partly responsible for the observed spontaneous CGP3 induction. Whereas previous studies focused mainly on the induction of prophages at the population level, we analyzed the spontaneous CGP3 induction at the single-cell level using promoters of phage genes (Pint2 and Plysin) fused to reporter genes encoding fluorescent proteins. Flow-cytometric analysis revealed a spontaneous CGP3 activity in about 0.01 to 0.08% of the cells grown in standard minimal medium, which displayed a significantly reduced viability. A PrecA-eyfp promoter fusion revealed that a small fraction of C. glutamicum cells (∼0.2%) exhibited a spontaneous induction of the SOS response. Correlation of PrecA to the activity of downstream SOS genes (PdivS and PrecN) confirmed a bona fide induction of this stress response rather than stochastic gene expression. Interestingly, the reporter output of PrecA and CGP3 promoter fusions displayed a positive correlation at the single-cell level (ρ = 0.44 to 0.77). Furthermore, analysis of the PrecA-eyfp/Pint2-e2-crimson strain during growth revealed the highest percentage of spontaneous PrecA and Pint2 activity in the early exponential phase, when fast replication occurs. Based on these studies, we postulate that spontaneously occurring DNA damage induces the SOS response, which in turn triggers the induction of lysogenic prophages. PMID:24163339

Estimating disturbance effects from military training using developmental instability and physiological measures of plant stress

USGS Publications Warehouse

Duda, J.J.; Freeman, D.C.; Brown, M.L.; Graham, J.H.; Krzysik, A.J.; Emlen, J.M.; Zak, J.C.; Kovacic, D.A.

2004-01-01

We used developmental instability, water potential, and variable fluorescence to determine if populations of winged sumac (Rhus copallinum) were being negatively effected by military training disturbance. We established nine sites that represented a land-use disturbance gradient with three impact levels (low, medium, and high), the effects mostly due to mechanized infantry training maneuvers. Although mean values of developmental instability, water potential, and variable fluorescence differed significantly among sites, the patterns did not consistently differentiate sites relative to the disturbance gradient. At the population level, some measures of developmental instability and variable fluorescence were positively correlated. All nine sites consisted of habitat mosaics, with the abundance of higher quality habitat patches and canopy gaps closely related to habitat impacts. It may be that R. copallinum is selecting similar micro-environments at all sites and therefore minimizing inter-site variation in stress measures, despite large differences in overall habitat condition. Our results call for caution in developing ecological indicators using the response of physiological and morphological measures from a single plant species. ?? 2003 Elsevier Ltd. All rights reserved.

Stoichiometry determination of (Pb,La)(Zr,Ti)O3-type nano-crystalline ferroelectric ceramics by wavelength-dispersive X-ray fluorescence spectrometry.

PubMed

Sitko, Rafał; Zawisza, Beata; Kita, Andrzej; Płońska, Małgorzata

2006-07-01

Analysis of small samples of lanthanum-doped lead zirconate titanate (PLZT) by wavelength-dispersive X-ray fluorescence spectrometry (WDXRF) is presented. The powdered material in ca. 30 mg was suspended in water and collected on the membrane filter. The pure oxide standards (PbO, La2O3, ZrO2 and TiO2) were used for calibration. The matrix effects were corrected using a theoretical influence coefficients algorithm for intermediate-thickness specimens. The results from XRF method were compared with the results from the inductively coupled plasma optical emission spectrometry (ICP-OES). Agreement between XRF and ICP-OES analysis was satisfactory and indicates the usefulness of XRF method for stoichiometry determination of PLZT.

Analysis of field-oriented controlled induction motor drives under sensor faults and an overview of sensorless schemes.

PubMed

Arun Dominic, D; Chelliah, Thanga Raj

2014-09-01

To obtain high dynamic performance on induction motor drives (IMD), variable voltage and variable frequency operation has to be performed by measuring speed of rotation and stator currents through sensors and fed back them to the controllers. When the sensors are undergone a fault, the stability of control system, may be designed for an industrial process, is disturbed. This paper studies the negative effects on a 12.5 hp induction motor drives when the field oriented control system is subjected to sensor faults. To illustrate the importance of this study mine hoist load diagram is considered as shaft load of the tested machine. The methods to recover the system from sensor faults are discussed. In addition, the various speed sensorless schemes are reviewed comprehensively. Copyright © 2014 ISA. Published by Elsevier Ltd. All rights reserved.

Hemodynamic responses to etomidate versus ketamine-thiopental sodium combination for anesthetic induction in coronary artery bypass graft surgery patients with low ejection fraction: a double-blind, randomized, clinical trial.

PubMed

Habibi, Mohammad Reza; Baradari, Afshin Gholipour; Soleimani, Aria; Emami Zeydi, Amir; Nia, Hamid Sharif; Habibi, Ali; Onagh, Naser

2014-10-01

During induction of anesthesia and intubation, hemodynamic changes are very important; especially in patients with coronary artery disease (CAD) and left ventricular dysfunction. A little information is available on the hemodynamic effects of a combination of ketamine-thiopental for induction of anesthesia in patients undergoing coronary artery bypass graft (CABG) surgery, with impaired ventricular function. The aim of this study was to compare the hemodynamic responses to etomidate versus ketamine-thiopental sodium combination for anesthetic induction in CABG surgery patients with low ejection fraction (EF<45%). In a double blind randomized clinical trial, a total of 100 patients, scheduled for elective CABG surgery were randomly assigned into two groups. These patients received either etomidate or ketamine-thiopental sodium combination at induction of anesthesia. Hemodynamics variable were measured and recorded at baseline, immediately before and after laryngoscopy and intubation, one, two and three minutes after intubation. Also, muscle twitching incidence among patients in two groups was evaluated. No significant differences between the two groups regarding the changes of hemodynamic variables including systolic and diastolic arterial blood pressure, mean arterial pressure and heart rate, were notice (p>0.05). Muscle twitching was not observed in the two groups. Hemodynamic stability after administration of ketamine-thiopental sodium combination for induction of anesthesia in patients undergoing CABG surgery, with impaired ventricular function, supports the clinical impression that this combination is safe in CABG surgery patients with low EF.

Reliable measurement of E. coli single cell fluorescence distribution using a standard microscope set-up.

PubMed

Cortesi, Marilisa; Bandiera, Lucia; Pasini, Alice; Bevilacqua, Alessandro; Gherardi, Alessandro; Furini, Simone; Giordano, Emanuele

2017-01-01

Quantifying gene expression at single cell level is fundamental for the complete characterization of synthetic gene circuits, due to the significant impact of noise and inter-cellular variability on the system's functionality. Commercial set-ups that allow the acquisition of fluorescent signal at single cell level (flow cytometers or quantitative microscopes) are expensive apparatuses that are hardly affordable by small laboratories. A protocol that makes a standard optical microscope able to acquire quantitative, single cell, fluorescent data from a bacterial population transformed with synthetic gene circuitry is presented. Single cell fluorescence values, acquired with a microscope set-up and processed with custom-made software, are compared with results that were obtained with a flow cytometer in a bacterial population transformed with the same gene circuitry. The high correlation between data from the two experimental set-ups, with a correlation coefficient computed over the tested dynamic range > 0.99, proves that a standard optical microscope- when coupled with appropriate software for image processing- might be used for quantitative single-cell fluorescence measurements. The calibration of the set-up, together with its validation, is described. The experimental protocol described in this paper makes quantitative measurement of single cell fluorescence accessible to laboratories equipped with standard optical microscope set-ups. Our method allows for an affordable measurement/quantification of intercellular variability, whose better understanding of this phenomenon will improve our comprehension of cellular behaviors and the design of synthetic gene circuits. All the required software is freely available to the synthetic biology community (MUSIQ Microscope flUorescence SIngle cell Quantification).

A Neuron-Based Screening Platform for Optimizing Genetically-Encoded Calcium Indicators

PubMed Central

Schreiter, Eric R.; Hasseman, Jeremy P.; Tsegaye, Getahun; Fosque, Benjamin F.; Behnam, Reza; Shields, Brenda C.; Ramirez, Melissa; Kimmel, Bruce E.; Kerr, Rex A.; Jayaraman, Vivek; Looger, Loren L.; Svoboda, Karel; Kim, Douglas S.

2013-01-01

Fluorescent protein-based sensors for detecting neuronal activity have been developed largely based on non-neuronal screening systems. However, the dynamics of neuronal state variables (e.g., voltage, calcium, etc.) are typically very rapid compared to those of non-excitable cells. We developed an electrical stimulation and fluorescence imaging platform based on dissociated rat primary neuronal cultures. We describe its use in testing genetically-encoded calcium indicators (GECIs). Efficient neuronal GECI expression was achieved using lentiviruses containing a neuronal-selective gene promoter. Action potentials (APs) and thus neuronal calcium levels were quantitatively controlled by electrical field stimulation, and fluorescence images were recorded. Images were segmented to extract fluorescence signals corresponding to individual GECI-expressing neurons, which improved sensitivity over full-field measurements. We demonstrate the superiority of screening GECIs in neurons compared with solution measurements. Neuronal screening was useful for efficient identification of variants with both improved response kinetics and high signal amplitudes. This platform can be used to screen many types of sensors with cellular resolution under realistic conditions where neuronal state variables are in relevant ranges with respect to timing and amplitude. PMID:24155972

Evaluation of mRNA expression levels and electrophysiological function of neuron-like cells derived from canine bone marrow stromal cells.

PubMed

Nakano, Rei; Edamura, Kazuya; Sugiya, Hiroshi; Narita, Takanori; Okabayashi, Ken; Moritomo, Tadaaki; Teshima, Kenji; Asano, Kazushi; Nakayama, Tomohiro

2013-10-01

To investigate the in vitro differentiation of canine bone marrow stromal cells (BMSCs) into functional, mature neurons. Bone marrow from 6 adult dogs. BMSCs were isolated from bone marrow and chemically induced to develop into neurons. The morphology of the BMSCs during neuronal induction was monitored, and immunocytochemical analyses for neuron markers were performed after the induction. Real-time PCR methods were used to evaluate the mRNA expression levels of markers for neural stem or progenitor cells, neurons, and ion channels, and western blotting was used to assess the expression of neuronal proteins before and after neuronal induction. The electrophysiological properties of the neuron-like cells induced from canine BMSCs were evaluated with fluorescent dye to monitor Ca(2)+ influx. Canine BMSCs developed a neuron-like morphology after neuronal induction. Immunocytochemical analysis revealed that these neuron-like cells were positive for neuron markers. After induction, the cells' mRNA expression levels of almost all neuron and ion channel markers increased, and the protein expression levels of nestin and neurofilament-L increased significantly. However, the neuron-like cells derived from canine BMSCs did not have the Ca(2)+ influx characteristic of spiking neurons. Although canine BMSCs had neuron-like morphological and biochemical properties after induction, they did not develop the electrophysiological characteristics of neurons. Thus, these results have suggested that canine BMSCs could have the capacity to differentiate into a neuronal lineage, but the differentiation protocol used may have been insufficient to induce development into functional neurons.

Wireless Monitoring of Induction Machine Rotor Physical Variables

PubMed Central

Doolan Fernandes, Jefferson; Carvalho Souza, Francisco Elvis; de Paiva, José Alvaro

2017-01-01

With the widespread use of electric machines, there is a growing need to extract information from the machines to improve their control systems and maintenance management. The present work shows the development of an embedded system to perform the monitoring of the rotor physical variables of a squirrel cage induction motor. The system is comprised of: a circuit to acquire desirable rotor variable(s) and value(s) that send it to the computer; a rectifier and power storage circuit that converts an alternating current in a continuous current but also stores energy for a certain amount of time to wait for the motor’s shutdown; and a magnetic generator that harvests energy from the rotating field to power the circuits mentioned above. The embedded system is set on the rotor of a 5 HP squirrel cage induction motor, making it difficult to power the system because it is rotating. This problem can be solved with the construction of a magnetic generator device to avoid the need of using batteries or collector rings and will send data to the computer using a wireless NRF24L01 module. For the proposed system, initial validation tests were made using a temperature sensor (DS18b20), as this variable is known as the most important when identifying the need for maintenance and control systems. Few tests have shown promising results that, with further improvements, can prove the feasibility of using sensors in the rotor. PMID:29156564

Wireless Monitoring of Induction Machine Rotor Physical Variables.

PubMed

Doolan Fernandes, Jefferson; Carvalho Souza, Francisco Elvis; Cipriano Maniçoba, Glauco George; Salazar, Andrés Ortiz; de Paiva, José Alvaro

2017-11-18

With the widespread use of electric machines, there is a growing need to extract information from the machines to improve their control systems and maintenance management. The present work shows the development of an embedded system to perform the monitoring of the rotor physical variables of a squirrel cage induction motor. The system is comprised of: a circuit to acquire desirable rotor variable(s) and value(s) that send it to the computer; a rectifier and power storage circuit that converts an alternating current in a continuous current but also stores energy for a certain amount of time to wait for the motor's shutdown; and a magnetic generator that harvests energy from the rotating field to power the circuits mentioned above. The embedded system is set on the rotor of a 5 HP squirrel cage induction motor, making it difficult to power the system because it is rotating. This problem can be solved with the construction of a magnetic generator device to avoid the need of using batteries or collector rings and will send data to the computer using a wireless NRF24L01 module. For the proposed system, initial validation tests were made using a temperature sensor (DS18b20), as this variable is known as the most important when identifying the need for maintenance and control systems. Few tests have shown promising results that, with further improvements, can prove the feasibility of using sensors in the rotor.

Effect of different methods of pulse width modulation on power losses in an induction motor

NASA Astrophysics Data System (ADS)

Gulyaev, Alexander; Fokin, Dmitrii; Shuharev, Sergey; Ten, Evgenii

2017-10-01

We consider the calculation of modulation power losses in a system “induction motor-inverter” for various pulse width modulation (PWM) methods of the supply voltage. Presented values of modulation power losses are the result of modeling a system “DC link - two-level three-phase voltage inverter - induction motor - load”. In this study the power losses in a system “induction motor - inverter” are computed, as well as losses caused by higher harmonics of PWM supply voltage, followed by definition of active power consumed by the DC link for a specified value mechanical power on the induction motor shaft. Mechanical power was determined by the rotation speed and the torque on the motor shaft in various quasi-sinusoidal supply voltage PWM modes. These calculations reveal the best coefficient of performance (COP) in a system of a variable frequency drive (VFD) with independent voltage inverter controlled by induction motor PWM.

Plant cell plasma membrane structure and properties under clinostatting

NASA Astrophysics Data System (ADS)

Polulakh, Yu. A.; Zhadko, S. I.; Klimchuk, D. A.; Baraboy, V. A.; Alpatov, A. N.; Sytnik, K. M.

Structural-functional organization of plasma membrane of pea roots seedling was investigated by methods of chemiluminescence, fluorescence probes, chromatography and freeze-fracture studies under normal conditions and clinostatting. Phase character of lipid peroxidation intensity was fixed. The initial phase of this process is characterized by lipid peroxidation decreasing with its next induction. The primary changes depending on free-radical mechanisms of lipid peroxidation were excellently revealed by chemiluminescence. Plasmalemma microviscosity increased on the average of 15-20 % under microgravity at the initial stages of its phenomenon. There were major changes of phosphatidilcholine and phosphatidilethanolamine contents. The total quantity of phospholipids remained rather stable. Changes of phosphatide acid concentration point to degradation and phospholipids biosynthesis. There were increases of unsaturated fatty acids mainly at the expense of linoleic and linolenic acids and also a decrease of saturated fatty acid content at the expense of palmitic and stearic acids. Unsaturation index of fatty acids increased as well. On the whole fatty acid composition was variable in comparison with phospholipids. Probably it is one of mechanisms of maintaining of microviscosity within definite limits. Considerable structural changes in organization of plasmalemma protein-lipid complex were not revealed by the freeze-fracture studies.

Demonstration of variable speed permanent magnet generator at small, low-head hydro site

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

Brown Kinloch, David

Small hydro developers face a limited set of bad choices when choosing a generator for a small low-head hydro site. Direct drive synchronous generators are expensive and technically complex to install. Simpler induction generators are higher speed, requiring a speed increaser, which results in inefficiencies and maintenance problems. In addition, both induction and synchronous generators turn at a fixed speed, causing the turbine to run off its peak efficiency curve whenever the available head is different than the designed optimum head.The solution to these problems is the variable speed Permanent Magnet Generators (PMG). At the Weisenberger Mill in Midway, KY,more » a variable speed Permanent Magnet Generator has been installed and demonstrated. This new PMG system replaced an existing induction generator that had a HTD belt drive speed increaser system. Data was taken from the old generator before it was removed and compared to data collected after the PMG system was installed. The new variable speed PMG system is calculated to produce over 96% more energy than the old induction generator system during an average year. This significant increase was primarily due to the PMG generator operating at the correct speed at the maximum head, and the ability for the PMG generator to reduce its speed to lower optimum speeds as the stream flow increased and the net head decreased.This demonstration showed the importance of being able to adjust the speed of fixed blade turbines. All fixed blade turbines with varying net heads could achieve higher efficiencies if the speed can be matched to the optimum speed as the head changes. In addition, this demonstration showed that there are many potential efficiencies that could be realized with variable speed technology at hydro sites where mismatched turbine and generator speeds result in lower power output, even at maximum head. Funding for this project came from the US Dept. of Energy, through Award Number DE-EE0005429.« less

Development of species-specific rDNA probes for Giardia by multiple fluorescent in situ hybridization combined with immunocytochemical identification of cyst wall antigens.

PubMed

Erlandsen, Stanley L; Jarroll, Edward; Wallis, Peter; van Keulen, Harry

2005-08-01

In this study, we describe the development of fluorescent oligonucleotide probes to variable regions in the small subunit of 16S rRNA in three distinct Giardia species. Sense and antisense probes (17-22 mer) to variable regions 1, 3, and 8 were labeled with digoxygenin or selected fluorochomes (FluorX, Cy3, or Cy5). Optimal results were obtained with fluorochome-labeled oligonucleotides for detection of rRNA in Giardia cysts. Specificity of fluorescent in situ hybridization (FISH) was shown using RNase digestion and high stringency to diminish the hybridization signal, and oligonucleotide probes for rRNA in Giardia lamblia, Giardia muris, and Giardia ardeae were shown to specifically stain rRNA only within cysts or trophozoites of those species. The fluorescent oligonucleotide specific for rRNA in human isolates of Giardia was positive for ten different strains. A method for simultaneous FISH detection of cysts using fluorescent antibody (genotype marker) and two oligonucleotide probes (species marker) permitted visualization of G. lamblia and G. muris cysts in the same preparation. Testing of an environmental water sample revealed the presence of FISH-positive G. lamblia cysts with a specific rDNA probe for rRNA, while negative cysts were presumed to be of animal or bird origin.

Dynamics of molecular excitons near a semiconductor surface studied by fluorescence quenching of polycrystalline tetracene on silicon

NASA Astrophysics Data System (ADS)

Piland, Geoffrey B.; Burdett, Jonathan J.; Hung, Tzu-Yao; Chen, Po-Hsun; Lin, Chi-Feng; Chiu, Tien-Lung; Lee, Jiun-Haw; Bardeen, Christopher J.

2014-05-01

Tetracene, a molecule that undergoes singlet fission, is deposited on Si with variable thickness LiF spacer layers. In agreement with earlier work (Hayashi et al., 1983 [10]), the fluorescence intensity of the tetracene greatly increases as the LiF thickness approaches 100 nm. This increase is partly due to a 30% increase in the prompt fluorescence decay time but mostly results from weaker coupling of the luminescence into the Si substrate. A decrease in the prompt fluorescence lifetime is observed as the tetracene thickness is increased on bare Si. We find no evidence for triplet energy transfer to the Si.

Inductive/Deductive Learning by Considering the Role of Gender--A Case Study of Iranian French-Learners

ERIC Educational Resources Information Center

Rahmatian, Rouhollah; Zarekar, Fatemeh

2016-01-01

This article defines the objective of discovering the first preferred styles of Iranian learners of French as a Foreign Language (FFL) as regards inductive or deductive learning; and secondly, the difference between gender-based learning tendencies. Considering these points as target variables, the questionnaire developed by Felder and Silverman…

Electrical Engineering and Nontechnical Design Variables of Multiple Inductive Loop Systems for Auditoriums

ERIC Educational Resources Information Center

Alterovitz, Gil

2004-01-01

This research analyzed both engineering and nontechnical issues involved in the use of Induction Loop Amplification (ILA) devices in auditoriums or large gathering places for hard-of-hearing individuals. A variety of parameters need to be taken into account to determine an optimal shape/configuration for the ILA device. In many cases, an optimal…

Using In-Situ Optical Sensors to Understand the Biogeochemistry of Dissolved Organic Matter Across a Stream Network

NASA Astrophysics Data System (ADS)

Wymore, Adam S.; Potter, Jody; Rodríguez-Cardona, Bianca; McDowell, William H.

2018-04-01

The advent of high-frequency in situ optical sensors provides new opportunities to study the biogeochemistry of dissolved organic matter (DOM) in aquatic ecosystems. We used fDOM (fluorescent dissolved organic matter) to examine the spatial and temporal variability in dissolved organic carbon (DOC) and dissolved organic nitrogen (DON) across a heterogeneous stream network that varies in NO3- concentration. Across the ten study streams fDOM explained twice the variability in the concentration of DOC (r2 = 0.82) compared to DON (r2 = 0.39), which suggests that the N-rich fraction of DOM is either more variable in its sources or more bioreactive than the more stable C-rich fraction. Among sites, DON molar fluorescence was approximately 3x more variable than DOC molar fluorescence and was correlated with changes in inorganic N, indicating that DON is both more variable in composition as well as highly responsive to changes in inorganic N. Laboratory results also indicate that the fDOM sensors we used perform as well as the excitation-emission wavelength pair generally referred to as the "tryptophan-like" peak when measured under laboratory conditions. However, since neither the field sensor not the laboratory measurements explained a large percentage of variation in DON concentrations, challenges still remain for monitoring the ambient pool of dissolved organic nitrogen. Sensor networks provide new insights into the potential reactivity of DOM and the variability in DOC and DON biogeochemistry across sites. These insights are needed to build spatially explicit models describing organic matter dynamics and water quality.

CDOM fluorescence as a proxy of DOC concentration in natural waters: a comparison of four contrasting tropical systems.

PubMed

Rochelle-Newall, E; Hulot, F D; Janeau, J L; Merroune, A

2014-01-01

Chromophoric dissolved organic matter (CDOM) fluorescence or absorption is often proposed as a rapid alternative to chemical methods for the estimation of bulk dissolved organic carbon (DOC) concentration in natural waters. However, the robustness of this method across a wide range of systems remains to be shown. We measured CDOM fluorescence and DOC concentration in four tropical freshwater and coastal environments (estuary and coastal, tropical shallow lakes, water from the freshwater lens of two small islands, and soil leachates). We found that although this method can provide an estimation of DOC concentration in sites with low variability in DOC and CDOM sources in systems where the variability of DOC and CDOM sources are high, this method should not be used as it will lead to errors in the estimation of the bulk DOC concentration.

Traffic Lights in Trichodesmium. Regulation of Photosynthesis for Nitrogen Fixation Studied by Chlorophyll Fluorescence Kinetic Microscopy1

PubMed Central

Küpper, Hendrik; Ferimazova, Naila; Šetlík, Ivan; Berman-Frank, Ilana

2004-01-01

We investigated interactions between photosynthesis and nitrogen fixation in the non-heterocystous marine cyanobacterium Trichodesmium IMS101 at the single-cell level by two-dimensional (imaging) microscopic measurements of chlorophyll fluorescence kinetics. Nitrogen fixation was closely associated with the appearance of cells with high basic fluorescence yield (F0), termed bright cells. In cultures aerated with normal air, both nitrogen fixation and bright cells appeared in the middle of the light phase. In cultures aerated with 5% oxygen, both processes occurred at a low level throughout most of the day. Under 50% oxygen, nitrogen fixation commenced at the beginning of the light phase but declined soon afterwards. Rapid reversible switches between fluorescence levels were observed, which indicated that the elevated F0 of the bright cells originates from reversible uncoupling of the photosystem II (PSII) antenna from the PSII reaction center. Two physiologically distinct types of bright cells were observed. Type I had about double F0 compared to the normal F0 in the dark phase and a PSII activity, measured as variable fluorescence (Fv = Fm − F0), similar to normal non-diazotrophic cells. Correlation of type I cells with nitrogen fixation, oxygen concentration, and light suggests that this physiological state is connected to an up-regulation of the Mehler reaction, resulting in oxygen consumption despite functional PSII. Type II cells had more than three times the normal F0 and hardly any PSII activity measurable by variable fluorescence. They did not occur under low-oxygen concentrations, but appeared under high-oxygen levels outside the diazotrophic period, suggesting that this state represents a reaction to oxidative stress not necessarily connected to nitrogen fixation. In addition to the two high-fluorescence states, cells were observed to reversibly enter a low-fluorescence state. This occurred mainly after a cell went through its bright phase and may represent a fluorescence-quenching recovery phase. PMID:15299119

Chromophoric dissolved organic matter (CDOM) variability in Barataria Basin using excitation-emission matrix (EEM) fluorescence and parallel factor analysis (PARAFAC).

PubMed

Singh, Shatrughan; D'Sa, Eurico J; Swenson, Erick M

2010-07-15

Chromophoric dissolved organic matter (CDOM) variability in Barataria Basin, Louisiana, USA,was examined by excitation emission matrix (EEM) fluorescence combined with parallel factor analysis (PARAFAC). CDOM optical properties of absorption and fluorescence at 355nm along an axial transect (36 stations) during March, April, and May 2008 showed an increasing trend from the marine end member to the upper basin with mean CDOM absorption of 11.06 + or - 5.01, 10.05 + or - 4.23, 11.67 + or - 6.03 (m(-)(1)) and fluorescence 0.80 + or - 0.37, 0.78 + or - 0.39, 0.75 + or - 0.51 (RU), respectively. PARAFAC analysis identified two terrestrial humic-like (component 1 and 2), one non-humic like (component 3), and one soil derived humic acid like (component 4) components. The spatial variation of the components showed an increasing trend from station 1 (near the mouth of basin) to station 36 (end member of bay; upper basin). Deviations from this increasing trend were observed at a bayou channel with very high chlorophyll-a concentrations especially for component 3 in May 2008 that suggested autochthonous production of CDOM. The variability of components with salinity indicated conservative mixing along the middle part of the transect. Component 1 and 4 were found to be relatively constant, while components 2 and 3 revealed an inverse relationship for the sampling period. Total organic carbon showed increasing trend for each of the components. An increase in humification and a decrease in fluorescence indices along the transect indicated an increase in terrestrial derived organic matter and reduced microbial activity from lower to upper basin. The use of these indices along with PARAFAC results improved dissolved organic matter characterization in the Barataria Basin. Copyright 2010 Elsevier B.V. All rights reserved.

Against Popperized epidemiology.

PubMed

Jacobsen, M

1976-03-01

The recommendation of Popper's philosophy of science should be adopted by epidemiologists is disputed. Reference is made to other authors who have shown that the most constructive elements in Popper's ideas have been advocated by earlier philosophers and have been used in epidemiology without abandoning inductive reasoning. It is argued that Popper's denigration of inductive methods is particularly harmful to epidemiology. Inductive reasoning and statistical inference play a key role in the science; it is suggested that unfamiliarity with these ideas contributes to widespread misunderstanding of the function of epidemiology. Attention is drawn to a common fallacy involving correlations between three random variables. The prevalence of the fallacy may be related to confusion between deductive and inductive logic.

Assessment of organic pollution of an industrial river by synchronous fluorescence and UV-vis spectroscopy: the Fensch River (NE France).

PubMed

Assaad, Aziz; Pontvianne, Steve; Pons, Marie-Noëlle

2017-05-01

To rapidly monitor the surface water quality in terms of organic pollution of an industrial river undergoing restoration, optical methods (UV-visible spectrometry and fluorescence) were applied in parallel to classical physical-chemical analyses. UV-visible spectra were analyzed using the maximum of the second derivative at 225 nm (related to nitrates), specific absorbance at 254 nm (SUVA 254 ), and the spectral slope between 275 and 295 nm (S 275-295 ) (related to the aromaticity and molecular weight of dissolved organic carbon). The synchronous fluorescence spectra (wavelength difference = 50 nm) exhibited a high variability in the composition of dissolved organic material between the upstream and downstream sections and also versus time. The principal components analysis of the entire set of synchronous fluorescence spectra helped to define three river sections with different pollution characteristics. Spectral decomposition was applied to the two most upstream sections: five fluorophores, classical in rivers impacted by domestic sewage and related to protein-like (λ ex  = 280 nm) and humic-like fluorescence (M-type with λ ex  ≈ 305-310 nm and C-type with λ ex  ≥ 335 nm), were identified. The irregular shape of the synchronous fluorescence spectra in the most downstream section is likely due to organic pollutants of industrial origin; however, their variability and the complexity of the spectra did not allow the further elucidation of their nature.

Direct Measurements of 3D Structure, Chemistry and Mass Density During the Induction Period of C3S Hydration

PubMed Central

Hu, Qinang; Aboustait, Mohammed; Kim, Taehwan; Ley, M. Tyler; Bullard, Jeffrey W.; Scherer, George; Hanan, Jay C.; Rose, Volker; Winarski, Robert; Gelb, Jeffrey

2017-01-01

The reasons for the start and end of the induction period of cement hydration remain topic of controversy. One long-standing hypothesis is that a thin metastable hydrate forming on the surface of cement grains significantly reduces the particle dissolution rate; the eventual disappearance of this layer re-establishes higher dissolution rates at the beginning of the acceleration period. However, the importance, or even the existence, of this metastable layer has been questioned because it cannot be directly detected in most experiments. In this work, a combined analysis using nano-tomography and nano-X-ray fluorescence makes the direct imaging of early hydration products possible. These novel X-ray imaging techniques provide quantitative measurements of 3D structure, chemical composition, and mass density of the hydration products during the induction period. This work does not observe a low density product on the surface of the particle, but does provide insights into the formation of etch pits and the subsequent hydration products that fill them. PMID:28919638

Nanoscale displacement sensing using microfabricated variable-inductance planar coils

NASA Astrophysics Data System (ADS)

Coskun, M. Bulut; Thotahewa, Kasun; Ying, York-Sing; Yuce, Mehmet; Neild, Adrian; Alan, Tuncay

2013-09-01

Microfabricated spiral inductors were employed for nanoscale displacement detection, suitable for use in implantable pressure sensor applications. We developed a variable inductor sensor consisting of two coaxially positioned planar coils connected in series to a measurement circuit. The devices were characterized by varying the air gap between the coils hence changing the inductance, while a Colpitts oscillator readout was used to obtain corresponding frequencies. Our approach shows significant advantages over existing methodologies combining a displacement resolution of 17 nm and low hysteresis (0.15%) in a 1 × 1 mm2 device. We show that resolution could be further improved by shrinking the device's lateral dimensions.

Variable frequency inverter for ac induction motors with torque, speed and braking control

NASA Technical Reports Server (NTRS)

Nola, F. J. (Inventor)

1975-01-01

A variable frequency inverter was designed for driving an ac induction motor which varies the frequency and voltage to the motor windings in response to varying torque requirements for the motor so that the applied voltage amplitude and frequency are of optimal value for any motor load and speed requirement. The slip frequency of the motor is caused to vary proportionally to the torque and feedback is provided so that the most efficient operating voltage is applied to the motor. Winding current surge is limited and a controlled negative slip causes motor braking and return of load energy to a dc power source.

A quantitative characterization of the yeast heterotrimeric G protein cycle

PubMed Central

Yi, Tau-Mu; Kitano, Hiroaki; Simon, Melvin I.

2003-01-01

The yeast mating response is one of the best understood heterotrimeric G protein signaling pathways. Yet, most descriptions of this system have been qualitative. We have quantitatively characterized the heterotrimeric G protein cycle in yeast based on direct in vivo measurements. We used fluorescence resonance energy transfer to monitor the association state of cyan fluorescent protein (CFP)-Gα and Gβγ-yellow fluorescent protein (YFP), and we found that receptor-mediated G protein activation produced a loss of fluorescence resonance energy transfer. Quantitative time course and dose–response data were obtained for both wild-type and mutant cells possessing an altered pheromone response. These results paint a quantitative portrait of how regulators such as Sst2p and the C-terminal tail of α-factor receptor modulate the kinetics and sensitivity of G protein signaling. We have explored critical features of the dynamics including the rapid rise and subsequent decline of active G proteins during the early response, and the relationship between the G protein activation dose–response curve and the downstream dose–response curves for cell-cycle arrest and transcriptional induction. Fitting the data to a mathematical model produced estimates of the in vivo rates of heterotrimeric G protein activation and deactivation in yeast. PMID:12960402

Responses of sun-induced chlorophyll fluorescence to biological and environmental variations measured with a versatile Fluorescence Auto-Measurement Equipment (FAME)

NASA Astrophysics Data System (ADS)

Gu, L.

2017-12-01

In this study, we examine responses of sun-induced chlorophyll fluorescence to biological and environmental variations measured with a versatile Fluorescence Auto-Measurement Equipment (FAME). FAME was developed to automatically and continuously measure chlorophyll fluorescence (F) of a leaf, plant or canopy in both laboratory and field environments, excited by either artificial light source or sunlight. FAME is controlled by a datalogger and allows simultaneous measurements of environmental variables complementary to the F signals. A built-in communication system allows FAME to be remotely monitored and data-downloaded. Radiance and irradiance calibrations can be done online. FAME has been applied in a variety of environments, allowing an investigation of biological and environmental controls on F emission.

Magnetic iron oxide modified pyropheophorbide-a fluorescence nanoparticles as photosensitizers for photodynamic therapy against ovarian cancer (SKOV-3) cells.

PubMed

Tan, Guanghui; Li, Wenting; Cheng, Jianjun; Wang, Zhiqiang; Wei, Shuquan; Jin, Yingxue; Guo, Changhong; Qu, Fengyu

2016-11-30

Magnetic iron oxide modified pyropheophorbide-a fluorescence nanoparticles, Fe 3 O 4 @SiO 2 @APTES@PPa (FSAP), were designed as magnetically targeted photodynamic antineoplastic agents and prepared through continuous covalent chemical modification on the surface of Fe 3 O 4 nanoparticles. The properties of the intermediates and the final product were comprehensively characterized by transmission electron microscopy, powder X-ray diffraction analysis, Fourier transform infrared spectroscopy, vibrating sample magnetometry, zeta potential measurement, ultraviolet-visible absorption spectroscopy, fluorescence emission spectroscopy, and thermogravimetric analysis. In this work, we demonstrated the in vitro photodynamic therapy (PDT) of FSAP against ovarian cancer (SKOV-3) cells, which indicated that FSAP could be taken up successfully and showed low dark toxicity without irradiation, but remarkable phototoxicity after irradiation. Meanwhile, FSAP had showed good biocompatibility and low dark toxicity against normal cells in the biological experiments on mouse normal fibroblast cell lines (L929 cells). In addition, in the photochemical process of FSAP mediated photodynamic therapy, the Type-II photo-oxygenation process (generated singlet oxygen) played an important role in the induction of cell damage.

Lysosomal Interaction of Akt with Phafin2: A Critical Step in the Induction of Autophagy

PubMed Central

Matsuda-Lennikov, Mami; Suizu, Futoshi; Hirata, Noriyuki; Hashimoto, Manabu; Kimura, Kohki; Nagamine, Tadashi; Fujioka, Yoichiro; Ohba, Yusuke; Iwanaga, Toshihiko; Noguchi, Masayuki

2014-01-01

Autophagy is an evolutionarily conserved mechanism for the gross disposal of intracellular proteins in mammalian cells and dysfunction in this pathway has been associated with human disease. Although the serine threonine kinase Akt is suggested to play a role in this process, little is known about the molecular mechanisms by which Akt induces autophagy. Using a yeast two-hybrid screen, Phafin2 (EAPF or PLEKHF2), a lysosomal protein with a unique structure of N-terminal PH (pleckstrin homology) domain and C-terminal FYVE (Fab 1, YOTB, Vac 1, and EEA1) domain was found to interact with Akt. A sucrose gradient fractionation experiment revealed that both Akt and Phafin2 co-existed in the same lysosome enriched fraction after autophagy induction. Confocal microscopic analysis and BiFC analysis demonstrated that both Akt and Phafin2 accumulate in the lysosome after induction of autophagy. BiFC analysis using PtdIns (3)P interaction defective mutant of Phafin2 demonstrated that lysosomal accumulation of the Akt-Phafin2 complex and subsequent induction of autophagy were lysosomal PtdIns (3)P dependent events. Furthermore, in murine macrophages, both Akt and Phafin2 were required for digestion of fluorescent bacteria and/or LPS-induced autophagy. Taken together, these findings establish that lysosomal accumulation of Akt and Phafin2 is a critical step in the induction of autophagy via an interaction with PtdIns (3)P. PMID:24416124

Interspecific Potato Breeding Lines Display Differential Colonization Patterns and Induced Defense Responses after Ralstonia solanacearum Infection

PubMed Central

Ferreira, Virginia; Pianzzola, María J.; Vilaró, Francisco L.; Galván, Guillermo A.; Tondo, María L.; Rodriguez, María V.; Orellano, Elena G.; Valls, Marc; Siri, María I.

2017-01-01

Potato (Solanum tuberosum L.) is one of the main hosts of Ralstonia solanacearum, the causative agent of bacterial wilt. This plant pathogen bacteria produce asymptomatic latent infections that promote its global spread, hindering disease control. A potato breeding program is conducted in Uruguay based on the introgression of resistance from the wild native species S. commersonii Dun. Currently, several backcrosses were generated exploiting the high genetic variability of this wild species resulting in advanced interspecific breeding lines with different levels of bacterial wilt resistance. The overall aim of this work was to characterize the interaction of the improved potato germplasm with R. solanacearum. Potato clones with different responses to R. solanacearum were selected, and colonization, dissemination and multiplication patterns after infection were evaluated. A R. solanacearum strain belonging to the phylotype IIB-sequevar 1, with high aggressiveness on potato was genetically modified to constitutively generate fluorescence and luminescence from either the green fluorescence protein gene or lux operon. These reporter strains were used to allow a direct and precise visualization of fluorescent and luminescent cells in plant tissues by confocal microscopy and luminometry. Based on wilting scoring and detection of latent infections, the selected clones were classified as susceptible or tolerant, while no immune-like resistance response was identified. Typical wilting symptoms in susceptible plants were correlated with high concentrations of bacteria in roots and along the stems. Tolerant clones showed a colonization pattern restricted to roots and a limited number of xylem vessels only in the stem base. Results indicate that resistance in potato is achieved through restriction of bacterial invasion and multiplication inside plant tissues, particularly in stems. Tolerant plants were also characterized by induction of anatomical and biochemical changes after R. solanacearum infection, including hyperplasic activity of conductor tissue, tylose production, callose and lignin deposition, and accumulation of reactive oxygen species. This study highlights the potential of the identified tolerant interspecific potato clones as valuable genetic resources for potato-breeding programs and leads to a better understanding of resistance against R. solanacearum in potato. PMID:28894453

Facile preparation and formation mechanism of Sr2Si5N8:Eu2+ red-emitting phosphors

NASA Astrophysics Data System (ADS)

Wang, Yang; Wang, Yunli; Wang, Ming; Shao, Yiran; Zhu, Yingchun

2018-05-01

The red-emitting Sr2Si5N8:Eu2+ phosphors have been synthesized in a new facile process using (oxy)nitride precursors by inductive calcination under N2 atmosphere at ordinary pressure. Different from the prevailing methods, lower cost raw materials, simpler pretreatment, without harsh conditions and a shorter reaction time are achieved. It was found that red-emitting Sr2Si5N8:Eu2+ phosphors were synthesized with high crystallinity and purity after 1 h inductive calcination. The formation mechanism was characterized by XRD, SEM, TEM and Fluorescence microscopy. It was demonstrated that a hexagonal mesophase of Sr-doped α-Si3N4 was primarily formed in the reaction process, which transformed into the final product of the orthorhombic Sr2Si5N8:Eu2+ phosphors. During the reaction process, the color of the samples transforms from greenish-yellow to orange and eventually to red. The as-prepared phosphors have a wide excitation in the range of 250 ∼ 570 nm which matches blue light chips and give a red-light emission peaking at 610 nm. The results indicate a promising prospect for a simple, efficient and inexpensive way to prepare Sr2Si5N8:Eu2+ phosphors for blue/UV-based warm-white LEDs and other fluorescent applications.

Detergent Induction of HEK 293A Cell Membrane Permeability Measured under Quiescent and Superfusion Conditions Using Whole Cell Patch Clamp

PubMed Central

2015-01-01

Detergents have several biological applications but present cytotoxicity concerns, since they can solubilize cell membranes. Using the IonFlux 16, an ensemble whole cell planar patch clamp, we observed that anionic sodium dodecyl sulfate (SDS), cationic cetyltrimethylammonium bromide (CTAB), and cationic, fluorescent octadecyl rhodamine B (ORB) increased the membrane permeability of cells substantially within a second of exposure, under superfusion conditions. Increased permeability was irreversible for 15 min. At subsolubilizing detergent concentrations, patched cells showed increased membrane currents that reached a steady state and were intact when imaged using fluorescence microscopy. SDS solubilized cells at concentrations of 2 mM (2× CMC), while CTAB did not solubilize cells even at concentrations of 10 mM (1000× CMC). The relative activity for plasma membrane current induction was 1:20:14 for SDS, CTAB, and ORB, respectively. Under quiescent conditions, the relative ratio of lipid to detergent in cell membranes at the onset of membrane permeability was 1:7:5 for SDS, CTAB, and ORB, respectively. The partition constants (K) for SDS, CTAB, and ORB were 23000, 55000, and 39000 M–1, respectively. Combining the whole cell patch clamp data and XTT viability data, SDS ≤ 0.2 mM and CTAB and ORB ≤ 1 mM induced cell membrane permeability without causing acute toxicity. PMID:24548291

Patterns of [PSI+] aggregation allow insights into cellular organization of yeast prion aggregates

PubMed Central

Tyedmers, Jens

2012-01-01

The yeast prion phenomenon is very widespread and mounting evidence suggests that it has an impact on cellular regulatory mechanisms related to phenotypic responses to changing environments. Studying the aggregation patterns of prion amyloids during different stages of the prion life cycle is a first key step to understand major principles of how and where cells generate, organize and turn-over prion aggregates. The induction of the [PSI+] state involves the actin cytoskeleton and quality control compartments such as the Insoluble Protein Deposit (IPOD). An initially unstable transitional induction state can be visualized by overexpression of the prion determinant and displays characteristic large ring- and ribbon-shaped aggregates consisting of poorly fragmented bundles of very long prion fibrils. In the mature prion state, the aggregation pattern is characterized by highly fragmented, shorter prion fibrils that form aggregates, which can be visualized through tagging with fluorescent proteins. The number of aggregates formed varies, ranging from a single large aggregate at the IPOD to multiple smaller ones, depending on several parameters discussed. Aggregate units below the resolution of light microscopy that are detectable by fluorescence correlation spectroscopy are in equilibrium with larger aggregates in this stage and can mediate faithful inheritance of the prion state. Loss of the prion state is often characterized by reduced fragmentation of prion fibrils and fewer, larger aggregates. PMID:22449721

Analytical procedure for characterization of medieval wall-paintings by X-ray fluorescence spectrometry, laser ablation inductively coupled plasma mass spectrometry and Raman spectroscopy

NASA Astrophysics Data System (ADS)

Syta, Olga; Rozum, Karol; Choińska, Marta; Zielińska, Dobrochna; Żukowska, Grażyna Zofia; Kijowska, Agnieszka; Wagner, Barbara

2014-11-01

Analytical procedure for the comprehensive chemical characterization of samples from medieval Nubian wall-paintings by means of portable X-ray fluorescence (pXRF), laser ablation inductively coupled plasma mass spectrometry (LA-ICPMS) and Raman spectroscopy (RS) was proposed in this work. The procedure was used for elemental and molecular investigations of samples from archeological excavations in Nubia (modern southern Egypt and northern Sudan). Numerous remains of churches with painted decorations dated back to the 7th-14th century were excavated in the region of medieval kingdoms of Nubia but many aspects of this art and its technology are still unknown. Samples from the selected archeological sites (Faras, Old Dongola and Banganarti) were analyzed in the form of transfers (n = 26), small fragments collected during the excavations (n = 35) and cross sections (n = 15). XRF was used to collect data about elemental composition, LA-ICPMS allowed mapping of selected elements, while RS was used to get the molecular information about the samples. The preliminary results indicated the usefulness of the proposed analytical procedure for distinguishing the substances, from both the surface and sub-surface domains of the wall-paintings. The possibility to identify raw materials from the wall-paintings will be used in the further systematic, archeometric studies devoted to the detailed comparison of various historic Nubian centers.

Variable-frequency inverter controls torque, speed, and braking in ac induction motors

NASA Technical Reports Server (NTRS)

Nola, F. J.

1974-01-01

Dc to ac inverter provides optimum frequency and voltage to ac induction motor, in response to different motor-load and speed requirements. Inverter varies slip frequency of motor in proportion to required torque. Inverter protects motor from high current surges, controls negative slip to apply braking, and returns energy stored in momentum of load to dc power source.

Fuzzy – PI controller to control the velocity parameter of Induction Motor

NASA Astrophysics Data System (ADS)

Malathy, R.; Balaji, V.

2018-04-01

The major application of Induction motor includes the usage of the same in industries because of its high robustness, reliability, low cost, highefficiency and good self-starting capability. Even though it has the above mentioned advantages, it also have some limitations: (1) the standard motor is not a true constant-speed machine, itsfull-load slip varies less than 1 % (in high-horsepower motors).And (2) it is not inherently capable of providing variable-speedoperation. In order to solve the above mentioned problem smart motor controls and variable speed controllers are used. Motor applications involve non linearity features, which can be controlled by Fuzzy logic controller as it is capable of handling those features with high efficiency and it act similar to human operator. This paper presents individuality of the plant modelling. The fuzzy logic controller (FLC)trusts on a set of linguistic if-then rules, a rule-based Mamdani for closed loop Induction Motor model. Themotor model is designed and membership functions are chosenaccording to the parameters of the motor model. Simulation results contains non linearity in induction motor model. A conventional PI controller iscompared practically to fuzzy logic controller using Simulink.

Development of a new fluorescent reporter:operator system: location of AraC regulated genes in Escherichia coli K-12.

PubMed

Sellars, Laura E; Bryant, Jack A; Sánchez-Romero, María-Antonia; Sánchez-Morán, Eugenio; Busby, Stephen J W; Lee, David J

2017-08-03

In bacteria, many transcription activator and repressor proteins regulate multiple transcription units that are often distally distributed on the bacterial genome. To investigate the subcellular location of DNA bound proteins in the folded bacterial nucleoid, fluorescent reporters have been developed which can be targeted to specific DNA operator sites. Such Fluorescent Reporter-Operator System (FROS) probes consist of a fluorescent protein fused to a DNA binding protein, which binds to an array of DNA operator sites located within the genome. Here we have developed a new FROS probe using the Escherichia coli MalI transcription factor, fused to mCherry fluorescent protein. We have used this in combination with a LacI repressor::GFP protein based FROS probe to assess the cellular location of commonly regulated transcription units that are distal on the Escherichia coli genome. We developed a new DNA binding fluorescent reporter, consisting of the Escherichia coli MalI protein fused to the mCherry fluorescent protein. This was used in combination with a Lac repressor:green fluorescent protein fusion to examine the spatial positioning and possible co-localisation of target genes, regulated by the Escherichia coli AraC protein. We report that induction of gene expression with arabinose does not result in co-localisation of AraC-regulated transcription units. However, measurable repositioning was observed when gene expression was induced at the AraC-regulated promoter controlling expression of the araFGH genes, located close to the DNA replication terminus on the chromosome. Moreover, in dividing cells, arabinose-induced expression at the araFGH locus enhanced chromosome segregation after replication. Regions of the chromosome regulated by AraC do not colocalise, but transcription events can induce movement of chromosome loci in bacteria and our observations suggest a role for gene expression in chromosome segregation.

Factor VII R353Q genetic polymorphism is associated with altered warfarin sensitivity among CYP2C9 *1/*1 carriers.

PubMed

Mlynarsky, Liat; Bejarano-Achache, Idit; Muszkat, Mordechai; Caraco, Yoseph

2012-05-01

Warfarin responsiveness is characterized by marked interindividual variability. A major portion of this variability is attributed to CYP2C9 and VKORC1 polymorphisms, but almost 50% is still unaccounted for. This paper reports the first prospective study on the association between factor VII R353Q polymorphism and warfarin responsiveness during induction. Genotyping for factor VII R353Q and 323D/I polymorphisms was performed in a cohort consisting of 374 patients (198 CYP2C9*1/*1) treated with warfarin who were prospectively followed from warfarin initiation. Compared with *1/*1-R/R and *1/*1-R/Q genotype carriers, *1/*1-Q/Q homozygotes achieved higher International Normalized Ratio (INR) values while consuming lower warfarin doses. The greater sensitivity was illustrated by 82.1% higher Warfarin Sensitivity Index During Induction (WSIDI) (0.14 ± 0.11 vs. 0.08 ± 0.50 mg⁻¹ Mann-Whitney, P = 0.043). Multiple regression analysis consisting of both genetic and nongenetic factors explained 26% of WSIDI variability, with R353Q genetic polymorphism having a modest yet significant effect and accounting for 1.7% of the overall variability. Moreover, the incidence of overanticoagulation (i.e., INR > 4) was 6.94-fold higher among *1/*1-Q/Q vs. *1/*1-R/R&R/Q carriers during warfarin induction (Pearson chi-square, P = 0.005). These findings were not accounted for by a chance difference in the distribution of VKORC1 genotypes. Analysis of these parameters among the entire cohort, including CYP2C9*2 and CYP2C9*3 variant allele carriers, did not reach statistical significance. Warfarin responsiveness during induction was unrelated to factor VII 323D/I genetic polymorphism. The response to warfarin during induction is influenced by factor VII R353Q polymorphism. The prospective use of this polymorphism, along with CYP2C9 and VKORC1, may enhance the accuracy of warfarin loading. However, the impact of R353Q polymorphism on overall warfarin response is subtle, and it is therefore unlikely that its use would be of clinical importance.

Protochlorophyll complexes with similar steady-state fluorescence characteristics can differ in fluorescence lifetimes. A model study in Triton X-100.

PubMed

Myśliwa-Kurdziel, Beata; Solymosi, Katalin; Kruk, Jerzy; Böddi, Béla; Strzałka, Kazimierz

2007-03-01

The steady-state and time-resolved fluorescence characteristics of protochlorophyll (Pchl) dissolved in neat Triton X-100 and in Triton X-100 micelles were investigated, and the fluorescence lifetimes of different Pchl spectral forms were studied. Varying the concentration of Pchl or diluting the micellar solutions either with a buffer or with a micellar solution, 631-634, 645-655, 680-692 and above 700 nm emitting Pchl complexes were prepared, the ratios of which varied from one another. The fluorescence decay of the 631-634 nm emitting (monomeric) form had a mono-exponential character with a 5.4-ns fluorescence lifetime. The long-wavelength Pchl complexes (aggregates) had two fluorescence lifetime values within a range of 1.4-3.9 ns and 0.15-0.84 ns, which showed high variability in different environments. Depending on the conditions, either mono- or double-exponential fluorescence decay was found for a fluorescence band at 680-685 nm. These data show that despite their very similar steady-state fluorescence properties, Pchl complexes can differ in fluorescence lifetimes, which may reflect different molecular structures, intrinsic geometries or different molecular interactions. This underlines the importance of complex spectroscopic analysis for a precise description of native and artificial chlorophyllous pigment forms.

Heat generation and light scattering of green fluorescent protein-like pigments in coral tissue

NASA Astrophysics Data System (ADS)

Lyndby, Niclas H.; Kühl, Michael; Wangpraseurt, Daniel

2016-05-01

Green fluorescent protein (GFP)-like pigments have been proposed to have beneficial effects on coral photobiology. Here, we investigated the relationships between green fluorescence, coral heating and tissue optics for the massive coral Dipsastraea sp. (previously Favia sp.). We used microsensors to measure tissue scalar irradiance and temperature along with hyperspectral imaging and combined imaging of variable chlorophyll fluorescence and green fluorescence. Green fluorescence correlated positively with coral heating and scalar irradiance enhancement at the tissue surface. Coral tissue heating saturated for maximal levels of green fluorescence. The action spectrum of coral surface heating revealed that heating was highest under red (peaking at 680 nm) irradiance. Scalar irradiance enhancement in coral tissue was highest when illuminated with blue light, but up to 62% (for the case of highest green fluorescence) of this photon enhancement was due to green fluorescence emission. We suggest that GFP-like pigments scatter the incident radiation, which enhances light absorption and heating of the coral. However, heating saturates, because intense light scattering reduces the vertical penetration depth through the tissue eventually leading to reduced light absorption at high fluorescent pigment density. We conclude that fluorescent pigments can have a central role in modulating coral light absorption and heating.

Simulation experiments to elucidate variable fluorescence as a potential proxy for bulk microalgal viability from natural water, sediments and biofilms: Implication in ships ballast water management.

PubMed

Patil, Jagadish S; Anil, Arga Chandrashekar

2018-05-30

The variable fluorescence fluorometry measuring microalgal biomass (initial fluorescence - F 0 , a chl-a proxy) and photosynthetic efficiency (F v /F m ) has been suggested as a potential tool in ballast-water assessment. In ballast tank, microalgae can be found in contiguous compartments i.e., in water, sediment, and biofilms. Therefore the utility of F 0 and F v /F m depends upon proper background corrections, which is straightforward for water samples but not for sediment and biofilms. This study proposes procedures for correcting F 0 values from sediment and biofilms. Irrespective of the saturation flash protocol used on any sample types the outcome of the results from viable and non-viable microalgae will remain same. Stress experiments (continuous darkness and biocide treatments) confirm that variable fluorescence (F v ) can be used as a potential proxy for viable cells as the values were negligible for non-viable cells and increased with an increase in abundance. Through this study, the utility of F v and σ PSII (functional-absorption-cross-section of photosystem II) along with F 0 and F v /F m in providing additional information on cell-viability and algal-size group during assessment is discussed. The findings will have implications not only from the perspective of ballast water but also in testing/assays of specific interest (e.g. toxicity, water treatments, antifouling) and ecological studies involving microalgae. Copyright © 2018 Elsevier Ltd. All rights reserved.

Alpha Alumina Nanoparticle Conjugation to Cysteine Peptidase A and B: An Efficient Method for Autophagy Induction

PubMed Central

Beyzay, Fatemeh; Zavaran Hosseini, Ahmad; Soudi, Sara

2017-01-01

Background: Autophagy as a cellular pathway facilitates several immune responses against infection. It also eliminates invading pathogens through transferring content between the cytosol and the lysosomal vesicles and contributes to the cross-presentation of exogenous antigens to T lymphocytes via MHC class I pathway. Autophagy induction is one of the main targets for new drugs and future vaccine formulations. Nanoparticles are one of the candidates for autophagy induction. Cysteine Peptidase A (CPA) and Cysteine Peptidase B (CPB) are two members of papain family (Clan CA, family C1) enzyme that have been considered as a virulence factor of Leishmania (L.) major, making them suitable vaccine candidates. In this research, Leishmania major cysteine peptidase A and B (CPA and CPB) conjugation to alpha alumina nanoparticle was the main focus and their entrance efficacy to macrophages was assessed. Methods: For this purpose, CPA and CPB genes were cloned in expression vectors. Related proteins were extracted from transformed Escherichia coli (E. coli) and purified using Ni affinity column. Alpha alumina nanoparticles were conjugated to CPA/CPB proteins using Aldehyde/Hydrazine Reaction. Autophagy induction in macrophages was assessed using acridine orange staining. Results: CPA/CPB protein loading to nanoparticles was confirmed by Fourier Transform Infrared Spectroscopy. α-alumina conjugated CPA/CPB antigen uptake by macrophages at different concentrations was confirmed using fluorescence microscope and flowcytometry. Highly efficient CPA/CPB protein loading to α-alumina nanoparticles and rapid internalization to macrophages introduced these nanocarriers as a delivery tool. Acridine orange staining demonstrated higher autophagy induction in CPA/CPB protein conjugated with α-alumina nanoparticles. Conclusion: α-alumina nanoparticles may be a promising adjuvant in the development of therapeutic leishmania vaccines through antigen delivery to intracellular compartments, induction of autophagy and cross presentation to CD8 lymphocytes. PMID:28496946

Time-dependent upregulation of electron transport with concomitant induction of regulated excitation dissipation in Haslea diatoms.

PubMed

Perkins, R; Williamson, C; Lavaud, J; Mouget, J-L; Campbell, D A

2018-04-16

Photoacclimation by strains of Haslea "blue" diatom species H. ostrearia and H. silbo sp. nov. ined. was investigated with rapid light curves and induction-recovery curves using fast repetition rate fluorescence. Cultures were grown to exponential phase under 50 µmol m -2  s -1 photosynthetic available radiation (PAR) and then exposed to non-sequential rapid light curves where, once electron transport rate (ETR) had reached saturation, light intensity was decreased and then further increased prior to returning to near growth light intensity. The non-sequential rapid light curve revealed that ETR was not proportional to the instantaneously applied light intensity, due to rapid photoacclimation. Changes in the effective absorption cross sections for open PSII reaction centres (σ PSII ') or reaction centre connectivity (ρ) did not account for the observed increases in ETR under extended high light. σ PSII ' in fact decreased as a function of a time-dependent induction of regulated excitation dissipation Y(NPQ), once cells were at or above a PAR coinciding with saturation of ETR. Instead, the observed increases in ETR under extended high light were explained by an increase in the rate of PSII reopening, i.e. Q A - oxidation. This acceleration of electron transport was strictly light dependent and relaxed within seconds after a return to low light or darkness. The time-dependent nature of ETR upregulation and regulated NPQ induction was verified using induction-recovery curves. Our findings show a time-dependent induction of excitation dissipation, in parallel with very rapid photoacclimation of electron transport, which combine to make ETR independent of short-term changes in PAR. This supports a selective advantage for these diatoms when exposed to fluctuating light in their environment.

Predictability and environmental drivers of chlorophyll fluctuations vary across different time scales and regions of the North Sea

NASA Astrophysics Data System (ADS)

Blauw, Anouk N.; Benincà, Elisa; Laane, Remi W. P. M.; Greenwood, Naomi; Huisman, Jef

2018-02-01

Phytoplankton concentrations display strong temporal variability at different time scales. Recent advances in automated moorings enable detailed investigation of this variability. In this study, we analyzed phytoplankton fluctuations at four automated mooring stations in the North Sea, which measured phytoplankton abundance (chlorophyll) and several environmental variables at a temporal resolution of 12-30 min for two to nine years. The stations differed in tidal range, water depth and freshwater influence. This allowed comparison of the predictability and environmental drivers of phytoplankton variability across different time scales and geographical regions. We analyzed the time series using wavelet analysis, cross correlations and generalized additive models to quantify the response of chlorophyll fluorescence to various environmental variables (tidal and meteorological variables, salinity, suspended particulate matter, nitrate and sea surface temperature). Hour-to-hour and day-to-day fluctuations in chlorophyll fluorescence were substantial, and mainly driven by sinking and vertical mixing of phytoplankton cells, horizontal transport of different water masses, and non-photochemical quenching of the fluorescence signal. At the macro-tidal stations, these short-term phytoplankton fluctuations were strongly driven by the tides. Along the Dutch coast, variation in salinity associated with the freshwater influence of the river Rhine played an important role, while in the central North Sea variation in weather conditions was a major determinant of phytoplankton variability. At time scales of weeks to months, solar irradiance, nutrient conditions and thermal stratification were the dominant drivers of changes in chlorophyll concentrations. These results show that the dominant drivers of phytoplankton fluctuations differ across marine environments and time scales. Moreover, our findings show that phytoplankton variability on hourly to daily time scales should not be dismissed as environmental noise, but is related to vertical and horizontal particle transport driven by winds and tides. Quantification of these transport processes contributes to an improved predictability of marine phytoplankton concentrations.

Fluorescence and fluorescence-lifetime imaging microscopy (FLIM) to characterize yeast strains by autofluorescence

NASA Astrophysics Data System (ADS)

Bhatta, H.; Goldys, E. M.; Ma, J.

2006-02-01

We characterised populations of wild type baking and brewing yeast cells using intrinsic fluorescence and fluorescence lifetime microscopy, in order to obtain quantitative identifiers of different strains. The cell autofluorescence was excited at 405 nm and observed within 440-540 nm range where strong cell to cell variability was observed. The images were analyzed using customised public domain software, which provided information on cell size, intensity and texture-related features. In light of significant diversity of the data, statistical methods were utilized to assess the validity of the proposed quantitative identifiers for strain differentiation. The Kolmogorov-Smirnov test was applied to confirm that empirical distribution functions for size, intensity and entropy for different strains were statistically different. These characteristics were followed with culture age of 24, 48 and 72 h, (the latter corresponding to a stationary growth phase) and size, and to some extent entropy, were found to be independent of age. The fluorescence intensity presented a distinctive evolution with age, different for each of the examined strains. The lifetime analysis revealed a short decay time component of 1.4 ns and a second, longer one with the average value of 3.5 ns and a broad distribution. High variability of lifetime values within cells was observed however a lifetime texture feature in the studied strains was statistically different.

A dark green fluorescent protein as an acceptor for measurement of Förster resonance energy transfer.

PubMed

Murakoshi, Hideji; Shibata, Akihiro C E; Nakahata, Yoshihisa; Nabekura, Junichi

2015-10-15

Measurement of Förster resonance energy transfer by fluorescence lifetime imaging microscopy (FLIM-FRET) is a powerful method for visualization of intracellular signaling activities such as protein-protein interactions and conformational changes of proteins. Here, we developed a dark green fluorescent protein (ShadowG) that can serve as an acceptor for FLIM-FRET. ShadowG is spectrally similar to monomeric enhanced green fluorescent protein (mEGFP) and has a 120-fold smaller quantum yield. When FRET from mEGFP to ShadowG was measured using an mEGFP-ShadowG tandem construct with 2-photon FLIM-FRET, we observed a strong FRET signal with low cell-to-cell variability. Furthermore, ShadowG was applied to a single-molecule FRET sensor to monitor a conformational change of CaMKII and of the light oxygen voltage (LOV) domain in HeLa cells. These sensors showed reduced cell-to-cell variability of both the basal fluorescence lifetime and response signal. In contrast to mCherry- or dark-YFP-based sensors, our sensor allowed for precise measurement of individual cell responses. When ShadowG was applied to a separate-type Ras FRET sensor, it showed a greater response signal than did the mCherry-based sensor. Furthermore, Ras activation and translocation of its effector ERK2 into the nucleus could be observed simultaneously. Thus, ShadowG is a promising FLIM-FRET acceptor.

Induction of food craving experience: the role of mental imagery, dietary restraint, mood and coping strategies.

PubMed

Jáuregui-Lobera, I; Bolaños-Ríos, P; Valero, E; Ruiz Prieto, I

2012-01-01

Food craving consists of a strong motivational state whereby a person is driven to seek and ingest a specifically desired food. To explore the influence of mental imagery on the food craving experience as well as to analyse the role of different psychological variables. Participants consisted of 65 normal weight undergraduate students. An experimental induction of food craving was analysed considering the actual previous craving and the induced one as a state food craving. Measures of trait food craving, imaging ability, dietary restraint, anxiety, depression, and coping strategies were considered. Sweet foods in general and chocolate in particular were the most craved foods. During the induction thoughts and images were the most highly rated triggers, and all the different sensory modalities were involved. Anxiety, depression, and negative coping strategies influenced the results with regards to the food craving. This study confirms the role of mental imagery, the correlation between state and trait food craving, and the influence of different psychological variables on the food craving.

Method and apparatus for monitoring the rotating frequency of de-energized induction motors

DOEpatents

Mikesell, H.E.; Lucy, E.

1998-02-03

The rotational speed of a coasting induction motor is measured by sensing e residual electrical voltages at the power terminals of the motor, thus eliminating the need for conventional tachometer equipment, additional mechanical components or modifications to the induction motor itself. The power terminal voltage signal is detected and transformed into a DC voltage proportional to the frequency of the signal. This DC voltage can be input to the control system of a variable frequency motor controller to regulate the output characteristics thereof relative to the speed of the coasting motor. 6 figs.

Method and apparatus for monitoring the rotating frequency of de-energized induction motors

DOEpatents

Mikesell, Harvey E.; Lucy, Eric

1998-01-01

The rotational speed of a coasting induction motor is measured by sensing e residual electrical voltages at the power terminals of the motor, thus eliminating the need for conventional tachometer equipment, additional mechanical components or modifications to the induction motor itself. The power terminal voltage signal is detected and transformed into a DC voltage proportional to the frequency of the signal. This DC voltage can be input to the control system of a variable frequency motor controller to regulate the output characteristics thereof relative to the speed of the coasting motor.

Contrasting effect of dark-chilling on chloroplast structure and arrangement of chlorophyll-protein complexes in pea and tomato: plants with a different susceptibility to non-freezing temperature.

PubMed

Garstka, Maciej; Venema, Jan Henk; Rumak, Izabela; Gieczewska, Katarzyna; Rosiak, Malgorzata; Koziol-Lipinska, Joanna; Kierdaszuk, Borys; Vredenberg, Wim J; Mostowska, Agnieszka

2007-10-01

The effect of dark-chilling and subsequent photoactivation on chloroplast structure and arrangements of chlorophyll-protein complexes in thylakoid membranes was studied in chilling-tolerant (CT) pea and in chilling-sensitive (CS) tomato. Dark-chilling did not influence chlorophyll content and Chl a/b ratio in thylakoids of both species. A decline of Chl a fluorescence intensity and an increase of the ratio of fluorescence intensities of PSI and PSII at 120 K was observed after dark-chilling in thylakoids isolated from tomato, but not from pea leaves. Chilling of pea leaves induced an increase of the relative contribution of LHCII and PSII fluorescence. A substantial decrease of the LHCII/PSII fluorescence accompanied by an increase of that from LHCI/PSI was observed in thylakoids from chilled tomato leaves; both were attenuated by photoactivation. Chlorophyll fluorescence of bright grana discs in chloroplasts from dark-chilled leaves, detected by confocal laser scanning microscopy, was more condensed in pea but significantly dispersed in tomato, compared with control samples. The chloroplast images from transmission-electron microscopy revealed that dark-chilling induced an increase of the degree of grana stacking only in pea chloroplasts. Analyses of O-J-D-I-P fluorescence induction curves in leaves of CS tomato before and after recovery from chilling indicate changes in electron transport rates at acceptor- and donor side of PS II and an increase in antenna size. In CT pea leaves these effects were absent, except for a small but irreversible effect on PSII activity and antenna size. Thus, the differences in chloroplast structure between CS and CT plants, induced by dark-chilling are a consequence of different thylakoid supercomplexes rearrangements.

Molecular Beacon-Based MicroRNA Imaging During Neurogenesis.

PubMed

Lee, Jonghwan; Kim, Soonhag

2016-01-01

The fluorescence monitoring system for examining endogenous microRNA (miRNA) activity in cellular level provides crucial information on not only understanding a critical role of miRNA involving a variety of biological processes, but also evaluating miRNA expression patterns in a noninvasive manner. In this protocol, we report the details of a new procedure for a molecular beacon-based miRNA monitoring system, which includes the illustration scheme for miRNA detection strategy, exogenous miRNA detection, and measurement of endogenous miRNA expression level during neurogenesis. The fluorescence signal of miR-124a beacon quenched by BHQ2 was gradually recovered as increasing concentration of the miR-124a in tube. The functional work of miR-124a beacon was examined in intracellular environment, allowing for the internalization of the miR-124a beacon by lipofectamine, which resulted in activated fluorescent signals of the miR-124a beacon in the HeLa cells after the addition of synthetic miR-124a. The endogenous miR-124a expression level was detected by miR-124a beacon system during neurogenesis, showing brighter fluorescence intensity in cytoplasmic area of P19 cells after induction of neuronal differentiation by retinoic acid. The molecular beacon based-miRNA detection technique could be applicable to the simultaneous visualization of a variety of miRNA expression patterns using different fluorescence dyes. For the study of examining endogenous miRNA expression level using miRNA-beacon system, if cellular differentiation step is already prepared, transfection step of miR-124a beacon into P19 cells, and acquisition of activated fluorescence signal measured by confocal microscope can be conducted approximately within 6 h.

A Variable Frequency, Mis-Match Tolerant, Inductive Plasma Source

NASA Astrophysics Data System (ADS)

Rogers, Anthony; Kirchner, Don; Skiff, Fred

2014-10-01

Presented here is a survey and analysis of an inductively coupled, magnetically confined, singly ionized Argon plasma generated by a square-wave, variable frequency plasma source. The helicon-style antenna is driven directly by the class ``D'' amplifier without matching network for increased efficiency while maintaining independent control of frequency and applied power at the feed point. The survey is compared to similar data taken using a traditional exciter--power amplifier--matching network source. Specifically, the flexibility of this plasma source in terms of the independent control of electron plasma temperature and density is discussed in comparison to traditional source arrangements. Supported by US DOE Grant DE-FG02-99ER54543.

Classification of California streams using combined deductive and inductive approaches: Setting the foundation for analysis of hydrologic alteration

USGS Publications Warehouse

Pyne, Matthew I.; Carlisle, Daren M.; Konrad, Christopher P.; Stein, Eric D.

2017-01-01

Regional classification of streams is an early step in the Ecological Limits of Hydrologic Alteration framework. Many stream classifications are based on an inductive approach using hydrologic data from minimally disturbed basins, but this approach may underrepresent streams from heavily disturbed basins or sparsely gaged arid regions. An alternative is a deductive approach, using watershed climate, land use, and geomorphology to classify streams, but this approach may miss important hydrological characteristics of streams. We classified all stream reaches in California using both approaches. First, we used Bayesian and hierarchical clustering to classify reaches according to watershed characteristics. Streams were clustered into seven classes according to elevation, sedimentary rock, and winter precipitation. Permutation-based analysis of variance and random forest analyses were used to determine which hydrologic variables best separate streams into their respective classes. Stream typology (i.e., the class that a stream reach is assigned to) is shaped mainly by patterns of high and mean flow behavior within the stream's landscape context. Additionally, random forest was used to determine which hydrologic variables best separate minimally disturbed reference streams from non-reference streams in each of the seven classes. In contrast to stream typology, deviation from reference conditions is more difficult to detect and is largely defined by changes in low-flow variables, average daily flow, and duration of flow. Our combined deductive/inductive approach allows us to estimate flow under minimally disturbed conditions based on the deductive analysis and compare to measured flow based on the inductive analysis in order to estimate hydrologic change.

Hybrid renewable energy system using doubly-fed induction generator and multilevel inverter

NASA Astrophysics Data System (ADS)

Ahmed, Eshita

The proposed hybrid system generates AC power by combining solar and wind energy converted by a doubly-fed induction generator (DFIG). The DFIG, driven by a wind turbine, needs rotor excitation so the stator can supply a load or the grid. In a variable-speed wind energy system, the stator voltage and its frequency vary with wind speed, and in order to keep them constant, variable-voltage and variable-frequency rotor excitation is to be provided. A power conversion unit supplies the rotor, drawing power either from AC mains or from a PV panel depending on their availability. It consists of a multilevel inverter which gives lower harmonic distortion in the stator voltage. Maximum power point tracking techniques have been implemented for both wind and solar power. The complete hybrid renewable energy system is implemented in a PSIM-Simulink interface and the wind energy conversion portion is realized in hardware using dSPACE controller board.

Active Power and Flux Control of a Self-Excited Induction Generator for a Variable-Speed Wind Turbine Generation

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

Na, Woonki; Muljadi, Eduard; Leighty, Bill

A Self-Excited Induction Generation (SEIG) for a variable speed wind turbine generation(VS-WG) is normally considered to be a good candidate for implementation in stand-alone applications such as battery charging, hydrogenation, water pumping, water purification, water desalination, and etc. In this study, we have examined a study on active power and flux control strategies for a SEIG for a variable speed wind turbine generation. The control analysis for the proposed system is carried out by using PSCAD software. In the process, we can optimize the control design of the system, thereby enhancing and expediting the control design procedure for this application.more » With this study, this control design for a SEIG for VS-WG can become the industry standard for analysis and development in terms of SEIG.« less

Design of inductive sensors for tongue control system for computers and assistive devices.

PubMed

Lontis, Eugen R; Struijk, Lotte N S A

2010-07-01

The paper introduces a novel design of air-core inductive sensors in printed circuit board (PCB) technology for a tongue control system. The tongue control system provides a quadriplegic person with a keyboard and a joystick type of mouse for interaction with a computer or for control of an assistive device. Activation of inductive sensors was performed with a cylindrical, soft ferromagnetic material (activation unit). Comparative analysis of inductive sensors in PCB technology with existing hand-made inductive sensors was performed with respect to inductance, resistance, and sensitivity to activation when the activation unit was placed in the center of the sensor. Optimisation of the activation unit was performed in a finite element model. PCBs with air-core inductive sensors were manufactured in a 10 layers, 100 microm and 120 microm line width technology. These sensors provided quality signals that could drive the electronics of the hand-made sensors. Furthermore, changing the geometry of the sensors allowed generation of variable signals correlated with the 2D movement of the activation unit at the sensors' surface. PCB technology for inductive sensors allows flexibility in design, automation of production and ease of possible integration with supplying electronics. The basic switch function of the inductive sensor can be extended to two-dimensional movement detection for pointing devices.

FUZZY LOGIC BASED INTELLIGENT CONTROL OF A VARIABLE SPEED CAGE MACHINE WIND GENERATION SYSTEM

EPA Science Inventory

The paper describes a variable-speed wind generation system where fuzzy logic principles are used to optimize efficiency and enhance performance control. A squirrel cage induction generator feeds the power to a double-sided pulse width modulated converter system which either pump...

FUZZY LOGIC BASED INTELLIGENT CONTROL OF A VARIABLE SPEED CAGE MACHINE WIND GENERATION SYSTEM

EPA Science Inventory

The report gives results of a demonstration of the successful application of fuzzy logic to enhance the performance and control of a variable-speed wind generation system. A squirrel cage induction generator feeds the power to either a double-sided pulse-width modulation converte...

Identification of tissular origin of particles based on autofluorescence multispectral image analysis at the macroscopic scale

NASA Astrophysics Data System (ADS)

Corcel, Mathias; Devaux, Marie-Françoise; Guillon, Fabienne; Barron, Cécile

2017-06-01

Powders produced from plant materials are heterogeneous in relation to native plant heterogeneity, and during grinding, dissociation often occurred at the tissue scale. The tissue composition of powdery samples could be modified through dry fractionation diagrams and impact their end-uses properties. If tissue identification is often made on native plant structure, this characterization is not straightforward in destructured samples such powders. Taking advantage of the autofluorescence properties of cell wall components, multispectral image acquisition is envisioned to identify the tissular origin of particles. Images were acquired on maize stem sections and ground tissues isolated from the same stem by hand dissection. The variability in fluorescence intensity profiles was analysed using principal component analysis. The correspondence between fluorescence profiles and the different tissues observed in maize sections was assessed based on histology or known compositional heterogeneity. Similar variability was encountered in fluorescence profiles extracted from powder leading to the potential ability to predict tissular origin based on this autofluorescence multispectral signal.

Bacterial Microcolonies in Gel Beads for High-Throughput Screening of Libraries in Synthetic Biology.

PubMed

Duarte, José M; Barbier, Içvara; Schaerli, Yolanda

2017-11-17

Synthetic biologists increasingly rely on directed evolution to optimize engineered biological systems. Applying an appropriate screening or selection method for identifying the potentially rare library members with the desired properties is a crucial step for success in these experiments. Special challenges include substantial cell-to-cell variability and the requirement to check multiple states (e.g., being ON or OFF depending on the input). Here, we present a high-throughput screening method that addresses these challenges. First, we encapsulate single bacteria into microfluidic agarose gel beads. After incubation, they harbor monoclonal bacterial microcolonies (e.g., expressing a synthetic construct) and can be sorted according their fluorescence by fluorescence activated cell sorting (FACS). We determine enrichment rates and demonstrate that we can measure the average fluorescent signals of microcolonies containing phenotypically heterogeneous cells, obviating the problem of cell-to-cell variability. Finally, we apply this method to sort a pBAD promoter library at ON and OFF states.

Estimating weak ratiometric signals in imaging data. I. Dual-channel data.

PubMed

Broder, Josef; Majumder, Anirban; Porter, Erika; Srinivasamoorthy, Ganesh; Keith, Charles; Lauderdale, James; Sornborger, Andrew

2007-09-01

Ratiometric fluorescent indicators are becoming increasingly prevalent in many areas of biology. They are used for making quantitative measurements of intracellular free calcium both in vitro and in vivo, as well as measuring membrane potentials, pH, and other important physiological variables of interest to researchers in many subfields. Often, functional changes in the fluorescent yield of ratiometric indicators are small, and the signal-to-noise ratio (SNR) is of order unity or less. In particular, variability in the denominator of the ratio can lead to very poor ratio estimates. We present a statistical optimization method for objectively detecting and estimating ratiometric signals in dual-wavelength measurements of fluorescent, ratiometric indicators that improves on standard methods. With the use of an appropriate statistical model for ratiometric signals and by taking the pixel-pixel covariance of an imaging dataset into account, we are able to extract user-independent spatiotemporal information that retains high resolution in both space and time.

Production of recombinant protein by a novel oxygen-induced system in Escherichia coli.

PubMed

Baez, Antonino; Majdalani, Nadim; Shiloach, Joseph

2014-04-07

The SoxRS regulon of E. coli is activated in response to elevated dissolved oxygen concentration likely to protect the bacteria from possible oxygen damage. The soxS expression can be increased up to 16 fold, making it a possible candidate for recombinant protein expression. Compared with the existing induction approaches, oxygen induction is advantageous because it does not involve addition or depletion of growth factors or nutrients, addition of chemical inducers or temperature changes that can affect growth and metabolism of the producing bacteria. It also does not affect the composition of the growth medium simplifying the recovery and purification processes. The soxS promoter was cloned into the commercial pGFPmut3.1 plasmid creating pAB49, an expression vector that can be induced by increasing oxygen concentration. The efficiency and the regulatory properties of the soxS promoter were characterized by measuring the GFP expression when the culture dissolved oxygen concentration was increased from 30% to 300% air saturation. The expression level of recombinant GFP was proportional to the oxygen concentration, demonstrating that pAB49 is a controllable expression vector. A possible harmful effect of elevated oxygen concentration on the recombinant product was found to be negligible by determining the protein-carbonyl content and its specific fluorescence. By performing high density growth in modified LB medium, the cells were induced by increasing the oxygen concentration. After 3 hours at 300% air saturation, GFP fluorescence reached 109000 FU (494 mg of GFP/L), representing 3.4% of total protein, and the cell concentration reached 29.1 g/L (DW). Induction of recombinant protein expression by increasing the dissolved oxygen concentration was found to be a simple and efficient alternative expression strategy that excludes the use of chemical, nutrient or thermal inducers that have a potential negative effect on cell growth or the product recovery.

Processes Affecting Variability of Fluorescence Signals from Benthic Targets in Shallow Waters

DTIC Science & Technology

1997-09-30

processes in the Department of Chemistry at Brookhaven National Laboratory. The model organisms used are primarily cultured zooxanthellae obtained from...and closed (Fm) photosystem II reaction centers in the zooxanthellae isolated from the fire coral, Montipora. The short lifetime curve corresponds...individual zooxanthellae strains, is highly correlated Figure 2. The correlation between the average fluorescence lifetimes, calculated from a four

Messenger RNA Detection in Leukemia Cell lines by Novel Metal-Tagged in situ Hybridization using Inductively Coupled Plasma Mass Spectrometry.

PubMed

Ornatsky, Olga I; Baranov, Vladimir I; Bandura, Dmitry R; Tanner, Scott D; Dick, John

2006-01-01

Conventional gene expression profiling relies on using fluorescent detection of hybridized probes. Physical characteristics of fluorophores impose limitations on achieving a highly multiplex gene analysis of single cells. Our work demonstrates the feasibility of using metal-tagged in situ hybridization for mRNA detection by inductively coupled plasma mass spectrometry (ICP-MS). ICP-MS as an analytical detector has a number of unique and relevant properties: 1) metals and their stable isotopes generate non-overlapping distinct signals that can be detected simultaneously; 2) these signals can be measured over a wide dynamic range; 3) ICP-MS is quantitative and very sensitive. We used commercial antibodies conjugated to europium (Eu) and gold together with biotinylated oligonucleotide probes reacted with terbium-labeled streptavidin to demonstrate simultaneous mRNA and protein detection by ICP-MS in leukemia cells.

Messenger RNA Detection in Leukemia Cell lines by Novel Metal-Tagged in situ Hybridization using Inductively Coupled Plasma Mass Spectrometry

PubMed Central

Ornatsky, Olga I.; Baranov, Vladimir I.; Bandura, Dmitry R.; Tanner, Scott D.; Dick, John

2006-01-01

Conventional gene expression profiling relies on using fluorescent detection of hybridized probes. Physical characteristics of fluorophores impose limitations on achieving a highly multiplex gene analysis of single cells. Our work demonstrates the feasibility of using metal-tagged in situ hybridization for mRNA detection by inductively coupled plasma mass spectrometry (ICP-MS). ICP-MS as an analytical detector has a number of unique and relevant properties: 1) metals and their stable isotopes generate non-overlapping distinct signals that can be detected simultaneously; 2) these signals can be measured over a wide dynamic range; 3) ICP-MS is quantitative and very sensitive. We used commercial antibodies conjugated to europium (Eu) and gold together with biotinylated oligonucleotide probes reacted with terbium-labeled streptavidin to demonstrate simultaneous mRNA and protein detection by ICP-MS in leukemia cells. PMID:23662035

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

Liu, Qinghao; Qin, Yixian; Zhou, Lei

In this study, we confirmed the autophagy induced by porcine reproductive and respiratory syndrome virus (PRRSV) in permissive cells and investigated the role of autophagy in the replication of PRRSV. We first demonstrated that PRRSV infection significantly results in the increased double-membrane vesicles, the accumulation of LC3 fluorescence puncta, and the raised ratio of LC3-II/{beta}-actin, in MARC-145 cells. Then we discovered that induction of autophagy by rapamycin significantly enhances the viral titers of PRRSV, while inhibition of autophagy by 3-MA and silencing of LC3 gene by siRNA reduces the yield of PRRSV. The results showed functional autolysosomes can be formedmore » after PRRSV infection and the autophagosome-lysosome-fusion inhibitor decreases the virus titers. We also examined the induction of autophagy by PRRSV infection in pulmonary alveolar macrophages. These findings indicate that autophagy induced by PRRSV infection plays a role in sustaining the replication of PRRSV in host cells.« less

The spread of Ras activity triggered by activation of a single dendritic spine.

PubMed

Harvey, Christopher D; Yasuda, Ryohei; Zhong, Haining; Svoboda, Karel

2008-07-04

In neurons, individual dendritic spines isolate N-methyl-d-aspartate (NMDA) receptor-mediated calcium ion (Ca2+) accumulations from the dendrite and other spines. However, the extent to which spines compartmentalize signaling events downstream of Ca2+ influx is not known. We combined two-photon fluorescence lifetime imaging with two-photon glutamate uncaging to image the activity of the small guanosine triphosphatase Ras after NMDA receptor activation at individual spines. Induction of long-term potentiation (LTP) triggered robust Ca2+-dependent Ras activation in single spines that decayed in approximately 5 minutes. Ras activity spread over approximately 10 micrometers of dendrite and invaded neighboring spines by diffusion. The spread of Ras-dependent signaling was necessary for the local regulation of the threshold for LTP induction. Thus, Ca2+-dependent synaptic signals can spread to couple multiple synapses on short stretches of dendrite.

A hybrid segmentation approach for geographic atrophy in fundus auto-fluorescence images for diagnosis of age-related macular degeneration.

PubMed

Lee, Noah; Laine, Andrew F; Smith, R Theodore

2007-01-01

Fundus auto-fluorescence (FAF) images with hypo-fluorescence indicate geographic atrophy (GA) of the retinal pigment epithelium (RPE) in age-related macular degeneration (AMD). Manual quantification of GA is time consuming and prone to inter- and intra-observer variability. Automatic quantification is important for determining disease progression and facilitating clinical diagnosis of AMD. In this paper we describe a hybrid segmentation method for GA quantification by identifying hypo-fluorescent GA regions from other interfering retinal vessel structures. First, we employ background illumination correction exploiting a non-linear adaptive smoothing operator. Then, we use the level set framework to perform segmentation of hypo-fluorescent areas. Finally, we present an energy function combining morphological scale-space analysis with a geometric model-based approach to perform segmentation refinement of false positive hypo- fluorescent areas due to interfering retinal structures. The clinically apparent areas of hypo-fluorescence were drawn by an expert grader and compared on a pixel by pixel basis to our segmentation results. The mean sensitivity and specificity of the ROC analysis were 0.89 and 0.98%.

Using simulated fluorescence cell micrographs for the evaluation of cell image segmentation algorithms.

PubMed

Wiesmann, Veit; Bergler, Matthias; Palmisano, Ralf; Prinzen, Martin; Franz, Daniela; Wittenberg, Thomas

2017-03-18

Manual assessment and evaluation of fluorescent micrograph cell experiments is time-consuming and tedious. Automated segmentation pipelines can ensure efficient and reproducible evaluation and analysis with constant high quality for all images of an experiment. Such cell segmentation approaches are usually validated and rated in comparison to manually annotated micrographs. Nevertheless, manual annotations are prone to errors and display inter- and intra-observer variability which influence the validation results of automated cell segmentation pipelines. We present a new approach to simulate fluorescent cell micrographs that provides an objective ground truth for the validation of cell segmentation methods. The cell simulation was evaluated twofold: (1) An expert observer study shows that the proposed approach generates realistic fluorescent cell micrograph simulations. (2) An automated segmentation pipeline on the simulated fluorescent cell micrographs reproduces segmentation performances of that pipeline on real fluorescent cell micrographs. The proposed simulation approach produces realistic fluorescent cell micrographs with corresponding ground truth. The simulated data is suited to evaluate image segmentation pipelines more efficiently and reproducibly than it is possible on manually annotated real micrographs.

LCLS in—photon out: fluorescence measurement of neon using soft x-rays

DOE PAGES

Obaid, Razib; Buth, Christian; Dakovski, Georgi L.; ...

2018-01-09

Here, we measured the fluorescence photon yield of neon upon soft x-ray ionization (~1200 eV) from the x-ray free-electron laser at Linac Coherent Light Source, and demonstrated the usage of a grazing incidence spectrometer with a variable line spacing grating to perform x-ray fluorescence spectroscopy on a gas phase system. Our measurements also allowed us to estimate the focal size of the beam from the theoretical description developed, in terms of the rate equation approximation accounting for photoionization shake off of neutral neon and double auger decay of single core holes.

LCLS in—photon out: fluorescence measurement of neon using soft x-rays

NASA Astrophysics Data System (ADS)

Obaid, Razib; Buth, Christian; Dakovski, Georgi L.; Beerwerth, Randolf; Holmes, Michael; Aldrich, Jeff; Lin, Ming-Fu; Minitti, Michael; Osipov, Timur; Schlotter, William; Cederbaum, Lorenz S.; Fritzsche, Stephan; Berrah, Nora

2018-02-01

We measured the fluorescence photon yield of neon upon soft x-ray ionization (∼1200 eV) from the x-ray free-electron laser at Linac Coherent Light Source, and demonstrated the usage of a grazing incidence spectrometer with a variable line spacing grating to perform x-ray fluorescence spectroscopy on a gas phase system. Our measurements also allowed us to estimate the focal size of the beam from the theoretical description developed, in terms of the rate equation approximation accounting for photoionization shake off of neutral neon and double auger decay of single core holes.

Quantitation of CYP1A1 and 1B1 mRNA in polycyclic aromatic hydrocarbon-treated human T-47D and HepG2 cells by a modified bDNA assay using fluorescence detection.

PubMed

Wu, Susan J; Spink, David C; Spink, Barbara C; Kaminsky, Laurence S

2003-01-15

The quantitation of mRNA, essential for assessing mechanisms of enzyme regulation, is normally carried out using reverse transcriptase-polymerase chain reaction (RT-PCR). An alternative method uses a signal-amplification nucleic acid probe assay, which measures RNA directly by the QuantiGene Expression Kit and incorporates branched DNA technology from Bayer and luminometer-based readings of a chemilumigenic alkaline phosphatase substrate. To broaden the utility of this assay, we investigated substitution of a fluorescent substrate, 2'-(2-benzothiazol)-6'-hydroxybenzothiazole phosphate and a fluorometer, and applied the method to quantitation of CYP1A1 and 1B1 mRNA in human T-47D and HepG2 cells following induction by benzo[a]pyrene (B[a]P) and dibenzo[a,h]anthracene (DB[a,h]A). The fluorescence response increased linearly for 200 min without photobleaching and increased linearly (r2=0.997) up to at least 0.2 microg total RNA. The data revealed that at 0.5 and 1.0 microM inducing agent, the induction of CYP1A1 mRNA in HepG2 cells by DB[a,h]A exceeded that by B[a]P by 18- and 6-fold, respectively. In T-47D cells B[a]P induced CYP1A1 mRNA by 23-fold and CYP1B1 mRNA by 3.9-fold. A B[a]P cocontaminant in the environment, arsenite, did not affect B[a]P-induced levels of CYP1A1 or 1B1 mRNA in these cells. The modified analytical system provides a rapid-throughput, reproducible, and less labor-intensive method than RT-PCR for quantifying cellular mRNA levels.

Indications for chlororespiration in relation to light regime in the marine diatom Thalassiosira weissflogii.

PubMed

Dijkman, Nicole A; Kroon, Bernd M A

2002-04-01

The marine diatom Thalassiosira weissflogii was cultured under a light regime simulating the daily rise and fall of the sun. The light regime caused a daily cycle in non-photochemical quenching. Remarkable were the changes in fluorescence directly after a light-to-dark transition that occurred in addition to the changes induced by non-photochemical quenching. A transient non-photochemical reduction of PQ and of Q(A) was indicated by a transient increase in apparent F(o) and by changes in the shape of the fluorescence induction curve. The observed changes developed approximately the first 100-120 s after a light-to-dark transition and could be reversed by the application of far-red illumination. Chlororespiration is thought to cause the reduction of PQ and, as the PQ-pool is in equilibrium with Q(A), also a reduction of Q(A). The function and ecological relevance of chlororespiration are discussed.

Structural reorganization of the fungal endoplasmic reticulum upon induction of mycotoxin biosynthesis.

PubMed

Boenisch, Marike Johanne; Broz, Karen Lisa; Purvine, Samuel Owen; Chrisler, William Byron; Nicora, Carrie Diana; Connolly, Lanelle Reine; Freitag, Michael; Baker, Scott Edward; Kistler, Harold Corby

2017-03-13

Compartmentalization of metabolic pathways to particular organelles is a hallmark of eukaryotic cells. Knowledge of the development of organelles and attendant pathways under different metabolic states has been advanced by live cell imaging and organelle specific analysis. Nevertheless, relatively few studies have addressed the cellular localization of pathways for synthesis of fungal secondary metabolites, despite their importance as bioactive compounds with significance to medicine and agriculture. When triggered to produce sesquiterpene (trichothecene) mycotoxins, the endoplasmic reticulum (ER) of the phytopathogenic fungus Fusarium graminearum is reorganized both in vitro and in planta. Trichothecene biosynthetic enzymes accumulate in organized smooth ER with pronounced expansion at perinuclear- and peripheral positions. Fluorescence tagged trichothecene biosynthetic proteins co-localize with the modified ER as confirmed by co-fluorescence and co-purification with known ER proteins. We hypothesize that changes to the fungal ER represent a conserved process in specialized eukaryotic cells such as in mammalian hepatocytes and B-cells.

Near-Infrared Phosphorus-Substituted Rhodamine with Emission Wavelength above 700 nm for Bioimaging.

PubMed

Chai, Xiaoyun; Cui, Xiaoyan; Wang, Baogang; Yang, Fan; Cai, Yi; Wu, Qiuye; Wang, Ting

2015-11-16

Phosphorus has been successfully fused into a classic rhodamine framework, in which it replaces the bridging oxygen atom to give a series of phosphorus-substituted rhodamines (PRs). Because of the electron-accepting properties of the phosphorus moiety, which is due to effective σ*-π* interactions and strengthened by the inductivity of phosphine oxide, PR exhibits extraordinary long-wavelength fluorescence emission, elongating to the region above 700 nm, with bathochromic shifts of 140 and 40 nm relative to rhodamine and silicon-substituted rhodamine, respectively. Other advantageous properties of the rhodamine family, including high molar extinction coefficient, considerable quantum efficiency, high water solubility, pH-independent emission, great tolerance to photobleaching, and low cytotoxicity, stay intact in PR. Given these excellent properties, PR is desirable for NIR-fluorescence imaging in vivo. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

X-ray fluorescence analysis of K, Al and trace elements in chloroaluminate melts

NASA Astrophysics Data System (ADS)

Shibitko, A. O.; Abramov, A. V.; Denisov, E. I.; Lisienko, D. G.; Rebrin, O. I.; Bunkov, G. M.; Rychkov, V. N.

2017-09-01

Energy dispersive x-ray fluorescence spectrometry was applied to quantitative determination of K, Al, Cr, Fe and Ni in chloroaluminate melts. To implement the external standard calibration method, an unconventional way of samples preparation was suggested. A mixture of metal chlorides was melted in a quartz cell at 350-450 °C under a slightly excessive pressure of purified argon (99.999 %). The composition of the calibration samples (CSs) prepared was controlled by means of the inductively coupled plasma atomic emission spectrometry (ICP-AES). The optimal conditions for analytical lines excitation were determined, the analytes calibration curves were obtained. There was some influence of matrix effects in synthesized samples on the analytical signal of some elements. The CSs are to be stored in inert gas atmosphere. The precision, accuracy, and reproducibility factors of the quantitative chemical analysis were computed.

Measuring iron in the brain using quantitative susceptibility mapping and X-ray fluorescence imaging

PubMed Central

Zheng, Weili; Nichol, Helen; Liu, Saifeng; Cheng, Yu-Chung N.; Haacke, E. Mark

2013-01-01

Measuring iron content in the brain has important implications for a number of neurodegenerative diseases. Quantitative susceptibility mapping (QSM), derived from magnetic resonance images, has been used to measure total iron content in vivo and in post mortem brain. In this paper, we show how magnetic susceptibility from QSM correlates with total iron content measured by X-ray fluorescence (XRF) imaging and by inductively coupled plasma mass spectrometry (ICPMS). The relationship between susceptibility and ferritin iron was estimated at 1.10 ± 0.08 ppb susceptibility per μg iron/g wet tissue, similar to that of iron in fixed (frozen/thawed) cadaveric brain and previously published data from unfixed brains. We conclude that magnetic susceptibility can provide a direct and reliable quantitative measurement of iron content and that it can be used clinically at least in regions with high iron content. PMID:23591072

A fluorimetric study of the thorium-morin system

USGS Publications Warehouse

Milkey, R.G.; Fletcher, M.H.

1957-01-01

Thorium reacts with morin to yield a yellow complex that fluoresces when irradiated with ultraviolet light. The effect on the fluorescence of such variables as concentration of acid, alcohol, thorium, morin, and complex; time, temperature and wave length of exciting light are studied to determine experimental conditions yielding maximum fluorescence. The effects of Zr+4, Al+3, Fe+3, Ca+2 and La+3 are discussed. The fundamental relationships between light absorption and fluorescence are expressed in a general equation that applies to a three-component system when the fluorescence is measured in a transmission-type fluorimeter. This general equation is used to obtain an expression for the fluorescence of the thoriummorin system. Equations, derived from experimental data, relate both the fraction of thorium reacted to form complex and the fraction of unquenched fluorescence to the concentration of uncombined morin. These functions, when combined with the general equation, give an expression whichrelates the total net fluorescence to the amount of uncombined morin in the solution. This last equation can be used to determine the one region for the concentration of uncombined morin that gives maximum sensitivity for the system. Calculated standard curves are in good agreement with experimental curves.

A fluorimetric study of the thorium-morin system

USGS Publications Warehouse

Milkey, Robert G.; Fletcher, Mary H.

1956-01-01

Thorium reacts with morin to yield a yellow complex that fluoresces when irradiated with ultraviolet light. The effect on the fluorescence of such variable as concentration of acid, alcohol, thorium, morin, and complex; time, temperature, and wavelength of exciting light are studied to determine experimental conditions yielding maximum fluorescence. The effects of Zr4+, Al3+, Fe3+, Ca2+, and La3+ are discussed. The fundamental relationships between light absorption and fluorescence are expressed in a general equation which applied to a three-component system when the fluorescence is measured in a transmission-type fluorimeter. This general equation is used to obtain an expression for the fluorescence of the thorium-morin system. Equations, derived from experimental data, related both the fraction of thorium reacted to form complex and the fraction of unquenched fluorescence to the concentration of uncombined morin. These functions, when combined with the general equation, give an expression which relates the total net fluorescence to the amount of uncombined morin in the solution. This last equation can be used to determine the one region for the concentration of uncombined morin that gives maximum sensitivity for the system. Calculated standard curves are in excellent agreement with experimental curves.

Microplastic Size-Dependent Toxicity, Oxidative Stress Induction, and p-JNK and p-p38 Activation in the Monogonont Rotifer (Brachionus koreanus).

PubMed

Jeong, Chang-Bum; Won, Eun-Ji; Kang, Hye-Min; Lee, Min-Chul; Hwang, Dae-Sik; Hwang, Un-Ki; Zhou, Bingsheng; Souissi, Sami; Lee, Su-Jae; Lee, Jae-Seong

2016-08-16

In this study, we evaluated accumulation and adverse effects of ingestion of microplastics in the monogonont rotifer (Brachionus koreanus). The dependence of microplastic toxicity on particle size was investigated by measuring several in vivo end points and studying the ingestion and egestion using 0.05-, 0.5-, and 6-μm nonfunctionalized polystyrene microbeads. To identify the defense mechanisms activated in response to microplastic exposure, the activities of several antioxidant-related enzymes and the phosphorylation status of mitogen-activated protein kinases (MAPKs) were determined. Exposure to polystyrene microbeads of all sizes led to significant size-dependent effects, including reduced growth rate, reduced fecundity, decreased lifespan and longer reproduction time. Rotifers exposed to 6-μm fluorescently labeled microbeads exhibited almost no fluorescence after 24 h, while rotifers exposed to 0.05- and 0.5-μm fluorescently labeled microbeads displayed fluorescence until 48 h, suggesting that 6-μm microbeads are more effectively egested from B. koreanus than 0.05- or 0.5-μm microbeads. This observation provides a potential explanation for our findings that microbead toxicity was size-dependent and smaller microbeads were more toxic. In vitro tests revealed that antioxidant-related enzymes and MAPK signaling pathways were significantly activated in response to microplastic exposure in a size-dependent manner.

Decreased Photosystem II Core Phosphorylation in a Yellow-Green Mutant of Wheat Showing Monophasic Fluorescence Induction Curve.

PubMed Central

Giardi, M. T.; Kucera, T.; Briantais, J. M.; Hodges, M.

1995-01-01

In the present work we study the regulation of the distribution of the phosphorylated photosystem II (PSII) core populations present in grana regions of the thylakoids from several plant species. The heterogeneous nature of PSII core phosphorylation has previously been reported (M.T. Giardi, F. Rigoni, R. Barbato [1992] Plant Physiol 100: 1948-1954; M.T. Giardi [1993] Planta 190: 107-113). The pattern of four phosphorylated PSII core populations in the grana regions appears to be ubiquitous in higher plants. In the dark, at least two phosphorylated PSII core populations are always detected. A mutant of wheat (Triticum durum) that shows monophasic room-temperature photoreduction of the primary quinone electron acceptor of PSII as measured by chlorophyll fluorescence increase in the presence and absence of 3-(3,4-dichlorophenyl)-1,1-dimethylurea and by fluorescence upon flash illumination in intact leaves also lacks the usual distribution of phosphorylated PSII core populations. In this mutant, the whole PSII core population pattern is changed, probably due to altered threonine kinase activity, which leads to the absence of light-induced phosphorylation of CP43 and D2 proteins. The results, correlated to previous experiments in vivo, support the idea that the functional heterogeneity observed by fluorescence is correlated in part to the PSII protein phosphorylation in the grana. PMID:12228652

Laser Induced Chlorophyll Fluorescence Spectra of Cajanus Cajan L Plant Growing Under Cadmium Stress

NASA Astrophysics Data System (ADS)

Gopal, Ram; Pandey, J. K.

2010-06-01

Laser-induced Chlorophyll fluorescence (LICF) spectra of Cajanus cajan L leaves treated with different concentrations of Cd (0.05, 0.5 and 1 mM) are recorded at 10 and 20 days after first treatment of cadmium. LICF spectra are recorded in the region of 650-780 nm using violet diode laser (405 nm). LICF spectra of plant leaves show two maxima near 685 and 730nm. Fluorescence induction kinetics (FIK) curve are recorded at 685 and 730 nm with red diode laser (635 nm) for excitation. The fluorescence intensity ratios (FIR) F685/F730 are calculated from LICF spectra and vitality index (Rfd) are determined from FIK curve. FIR and Rfd value are good stress indicator of plant health. These parameters along with chlorophyll content are used to analyze the effect of Cd on wheat plants. The result indicates that higher concentrations of Cd hazardous for photosynthetic activity and health of Arhar plants. The lower concentration of 0.05 mM shows stimulatory response up to 10 days while after 20 days this concentration also shows inhibitory response. R. Gopal, K. B. Mishra, M. Zeeshan, S. M. Prasad, and M. M. Joshi Curr. Sci., 83, 880, 2002 K. B. Mishra and R. Gopal Int. J. Rem. Sen., 29, 157, 2008 R. Maurya, S. M. Prasad, and R. Gopal J. Photochem. Photobio. C: Photochem. Rev., 9, 29, 2008

Gadolinium and 5-Aminolevulinic Acid-induced Protoporphyrin IX Levels in Human Gliomas: An Ex Vivo Quantitative Study to Correlate Protoporphyrin IX Levels and Blood-Brain Barrier Breakdown

PubMed Central

Valdés, Pablo A.; Moses, Ziev B.; Kim, Anthony; Belden, Clifford J.; Wilson, Brian C.; Paulsen, Keith D.; Roberts, David W.; Harris, Brent T.

2012-01-01

In recent years, 5-aminolevulinic acid (ALA)-induced protoporphyrin IX (PpIX) fluorescence guidance has been used as a surgical adjunct to improve the extent of resection of gliomas. Exogenous administration of ALA prior to surgery leads to the accumulation of red fluorescent PpIX in tumor tissue that the surgeon can visualize and thereby discriminate between normal and tumor tissue. Selective accumulation of PpIX has been linked to numerous factors, of which blood-brain barrier (BBB) breakdown has been suggested to be a key factor. To test the hypothesis that PpIX concentration (CPpIX) positively correlates with gadolinium (Gd) concentrations (CGd), we performed ex vivo measurements of PpIX and of Gd using Inductively-Coupled Plasma Mass Spectrometry (ICP-MS) the latter as a quantitative biomarker of BBB breakdown; this was corroborated with immunohistochemistry of microvascular density in surgical biopsies of patients undergoing fluorescence guided surgery for glioma .We found positive correlations between CPpIX and CGd (r = 0.58, p < 0.0001), and between CPpIX and microvascular density (r = 0.55, p < 0.0001), suggesting a significant, yet limited association between BBB breakdown and ALA-induced PpIX fluorescence. To our knowledge, this is the first time that Gd measurements by ICP-MS have been used in human gliomas. PMID:22878664

Reasoning in people with obsessive-compulsive disorder.

PubMed

Simpson, Jane; Cove, Jennifer; Fineberg, Naomi; Msetfi, Rachel M; J Ball, Linden

2007-11-01

The aim of this study was to investigate the inductive and deductive reasoning abilities of people with obsessive-compulsive disorder (OCD). Following previous research, it was predicted that people with OCD would show different abilities on inductive reasoning tasks but similar abilities to controls on deductive reasoning tasks. A two-group comparison was used with both groups matched on a range of demographic variables. Where appropriate, unmatched variables were entered into the analyses as covariates. Twenty-three people with OCD and 25 control participants were assessed on two tasks: an inductive reasoning task (the 20-questions task) and a deductive reasoning task (a syllogistic reasoning task with a content-neutral and content-emotional manipulation). While no group differences emerged on several of the parameters of the inductive reasoning task, the OCD group did differ on one, and arguably the most important, parameter by asking fewer correct direct-hypothesis questions. The syllogistic reasoning task results were analysed using both correct response and conclusion acceptance data. While no main effects of group were evident, significant interactions indicated important differences in the way the OCD group reasoned with content neutral and emotional syllogisms. It was argued that the OCD group's patterns of response on both tasks were characterized by the need for more information, states of uncertainty, and doubt and postponement of a final decision.

Manipulation of heart rate variability can modify response to anger-inducing stimuli.

PubMed

Francis, Heather M; Penglis, Kathryn M; McDonald, Skye

2016-10-01

Research suggests that heart rate variability (HRV) is a physiological indicator of the flexibility of the autonomic nervous system and can provide an objective measure of an individual's ability to appropriately match emotional responses to environmental demands. The present study investigated whether angry response to emotional stimuli was related to HRV, and whether manipulation of HRV using biofeedback could change the anger response in a healthy adult population. Fifty-eight participants received HRV biofeedback (n = 29) or an active control condition (n = 29). HRV measures included standard deviation of normal-to-normal intervals (SDNN), low-frequency (LF) and high-frequency (HF) power, and was recorded across three sessions: baseline, training, and anger induction. The anger induction procedure resulted in increased subjective experience of anger, as well as physiological changes. The biofeedback group had higher HRV than active controls both during the training session (SDNN and LF HRV) and during anger induction (LF HRV). HRV during anger induction was significantly associated with self-reported emotional response for participants receiving biofeedback but not for active controls. Results provide support for HRV as an index of emotion regulation, specifically anger. Further research is needed to determine whether long-term HRV biofeedback can have a lasting effect on managing anger.

Red fluorescent biofilm: the thick, the old, and the cariogenic

PubMed Central

Volgenant, Catherine M.C.; Hoogenkamp, Michel A.; Buijs, Mark J.; Zaura, Egija; ten Cate, Jacob (Bob) M.; van der Veen, Monique H.

2016-01-01

Background Some dental plaque fluoresces red. The factors involved in this fluorescence are yet unknown. Objective The aim of this study was to assess systematically the effect of age, thickness, and cariogenicity on the extent of red fluorescence produced by in vitro microcosm biofilms. Design The effects of biofilm age and thickness on red fluorescence were tested in a constant depth film fermentor (CDFF) by growing biofilms of variable thicknesses that received a constant supply of defined mucin medium (DMM) and eight pulses of sucrose/day. The influence of cariogenicity on red fluorescence was tested by growing biofilm on dentin disks receiving DMM, supplemented with three or eight pulses of sucrose/day. The biofilms were analyzed at different time points after inoculation, up to 24 days. Emission spectra were measured using a fluorescence spectrophotometer (λexc405 nm) and the biofilms were photographed with a fluorescence camera. The composition of the biofilms was assessed using 454-pyrosequecing of the 16S rDNA gene. Results From day 7 onward, the biofilms emitted increasing intensities of red fluorescence as evidenced by the combined red fluorescence peaks. The red fluorescence intensity correlated with biofilm thickness but not in a linear way. Biofilm fluorescence also correlated with the imposed cariogenicity, evidenced by the induced dentin mineral loss. Increasing the biofilm age or increasing the sucrose pulsing frequency led to a shift in the microbial composition. These shifts in composition were accompanied by an increase in red fluorescence. Conclusions The current study shows that a thicker, older, or more cariogenic biofilm results in a higher intensity of red fluorescence. PMID:27060056

Fluorescent pH sensor based on Ag@SiO2 core-shell nanoparticle.

PubMed

Bai, Zhenhua; Chen, Rui; Si, Peng; Huang, Youju; Sun, Handong; Kim, Dong-Hwan

2013-06-26

We have demonstrated a novel method for the preparation of a fluorescence-based pH sensor by combining the plasmon resonance band of Ag core and pH sensitive dye (HPTS). A thickness-variable silica shell is placed between Ag core and HPTS dye to achieve the maximum fluorescence enhancement. At the shell thickness of 8 nm, the fluorescence intensity increases 4 and 9 times when the sensor is excited at 405 and 455 nm, respectively. At the same time, the fluorescence intensity shows a good sensitivity toward pH value in the range of 5-9, and the ratio of emission intensity at 513 nm excited at 455 nm to that excited at 405 nm versus the pH value in the range of 5-9 is determined. It is believed that the present pH sensor has the potential for determining pH real time in the biological sample.

Segmentation and classification of cell cycle phases in fluorescence imaging.

PubMed

Ersoy, Ilker; Bunyak, Filiz; Chagin, Vadim; Cardoso, M Christina; Palaniappan, Kannappan

2009-01-01

Current chemical biology methods for studying spatiotemporal correlation between biochemical networks and cell cycle phase progression in live-cells typically use fluorescence-based imaging of fusion proteins. Stable cell lines expressing fluorescently tagged protein GFP-PCNA produce rich, dynamically varying sub-cellular foci patterns characterizing the cell cycle phases, including the progress during the S-phase. Variable fluorescence patterns, drastic changes in SNR, shape and position changes and abundance of touching cells require sophisticated algorithms for reliable automatic segmentation and cell cycle classification. We extend the recently proposed graph partitioning active contours (GPAC) for fluorescence-based nucleus segmentation using regional density functions and dramatically improve its efficiency, making it scalable for high content microscopy imaging. We utilize surface shape properties of GFP-PCNA intensity field to obtain descriptors of foci patterns and perform automated cell cycle phase classification, and give quantitative performance by comparing our results to manually labeled data.

Space fabrication demonstration system. [beam builder and induction fastening

NASA Technical Reports Server (NTRS)

1980-01-01

The development effort on the composite beam cap fabricator was completed within cost and close to abbreviated goals. The design and analysis of flight weight primary and secondary beam builder structures proceeded satisfactorily but remains curtailed until further funding is made available to complete the work. The induction fastening effort remains within cost and schedule constraints. Tests of the LARC prototype induction welder is continuing in an instrumented test stand comprised of a Dumore drill press (air over oil feed for variable applied loads) and a dynamometer to measure actual welding loads. Continued testing shows that the interface screening must be well impregnated with resin to ensure proper flow when bonding graphite-acrylic lap shear samples. Specimens prepared from 0.030 inch thick graphite-polyethersulfone are also available for future induction fastening evaluation.

Deep Space Gateway Ecosystem Observatory

NASA Astrophysics Data System (ADS)

Huemmrich, K. F.; Campbell, P. E.; Middleton, E. M.

2018-02-01

Advance global understanding of seasonal change and diurnal variability of terrestrial ecosystem function, photosynthesis, and stress responses using spectral reflectance, thermal, and fluorescence signals.

Micro-hole array fluorescent sensor based on AC-Dielectrophoresis (DEP) for simultaneous analysis of nano-molecules

NASA Astrophysics Data System (ADS)

Kim, Hye Jin; Kang, Dong-Hoon; Lee, Eunji; Hwang, Kyo Seon; Shin, Hyun-Joon; Kim, Jinsik

2018-02-01

We propose a simple fluorescent bio-chip based on two types of alternative current-dielectrophoretic (AC-DEP) force, attractive (positive DEP) and repulsive (negative DEP) force, for simultaneous nano-molecules analysis. Various radius of micro-holes on the bio-chip are designed to apply the different AC-DEP forces, and the nano-molecules are concentrated inside the micro-hole arrays according to the intensity of the DEP force. The bio-chip was fabricated by Micro Electro Mechanical system (MEMS) technique, and was composed of two layers; a SiO2 layer and Ta/Pt layer were accomplished for an insulation layer and a top electrode with micro-hole arrays to apply electric fields for DEP force, respectively. Each SiO2 and Ta/Pt layers were deposited by thermal oxidation and sputtering, and micro-hole arrays were fabricated with Inductively Coupled Plasma (ICP) etching process. For generation of each positive and negative DEP at micro-holes, we applied two types of sine-wave AC voltage with different frequency range alternately. The intensity of the DEP force was controlled by the radius of the micro-hole and size of nano-molecule, and calculated with COMSOL multi-physics. Three types of nano-molecules labelled with different fluorescent dye were used and the intensity of nano-molecules was examined by the fluorescent optical analysis after applying the DEP force. By analyzing the fluorescent intensities of the nano-molecules, we verify the various nano-molecules in analyte are located successfully inside corresponding micro-holes with different radius according to their size.

Primula auriculata Extracts Exert Cytotoxic and Apoptotic Effects against HT-29 Human Colon Adenocarcinoma Cells.

PubMed

Behzad, Sahar; Ebrahim, Karim; Mosaddegh, Mahmoud; Haeri, Ali

2016-01-01

Primula auriculata (Tootia) is one of the most important local medicinal plants in Hamedan district, Iran. To investigate cytotoxicity and apoptosis induction of crude methanolic extract and different fraction of it, we compared several methods on HT-29 human colon Adenocarcinoma cells. Cancer cell proliferation was measured by 3-(4, 5‑dimethylthiazolyl)2, 5‑diphenyl‑tetrazolium bromide (MTT) assay and apoptosis induction was analyzed by fluorescence microscopy (acridin orange/ethidium bromide, annexin V/propidium iodide staining, TUNEL assay and Caspase-3 activity assay). Crude methanolic extract (CM) inhibited the growth of malignant cells in a dose-dependent manner. Among solvent fractions, the dichloromethane fraction (CF) was found to be the most toxic compared to other fractions. With double staining methods, high percentage of 40 µg/mL of (CM) and (CF) treated cells exhibited typical characteristics of apoptotic cells. Apoptosis induction was also revealed by apoptotic fragmentation of nuclear DNA and activation of caspas-3 in treated cells. These findings indicate that crude methanolic extract and dichloromethan fraction of P.auriculata induced apoptosis and inhibited proliferation in colon cancer cells and could be used as a source for new lead structures in drug design to combat colon cancer.

Use of Bone Marrow-Derived Mesenchymal Stem Cells in Improving Thioacetamide Induced Liver Fibrosis in Rats

NASA Astrophysics Data System (ADS)

Mansour, Fatma A. A.; Shaheed, Iman; Hassan, Nabiha R. A.

Liver fibrosis, is one of big problems usually ends with cirrhosis which considered a life threatening disease as the only way of treatment is the liver transplantation, this study aimed to find a new way for fibrosis treatment by the use of bone marrow isolated Mesenchymal stem cells (MSCs). Thioacetamide (TAA) was used for fibrosis induction in male Sprague Dawely (SD) rats which divided into two random groups: group infused with TAA for fibrosis induction and group as control negative group. MSCs were isolated from bone marrow of twenty five (4-5) weeks male SD rats, and labeled with fluorescent material (PKH26) to confirm the homing of cells. After fibrosis induction, rats were divided into four subgroups to study the effect of MSCs injection in fibrosis treatment. After 4 weeks from MSCs administration, all rats were sacrificed. Liver tissue were collected for histopathological and immunohistopathological studies. In comparison with control groups, the treated groups with MSCs showed improvement in the amount of deposited collagen which decreased compared to control positive group. So MSCs can be used to replace liver transplantation in the treatment of fibrosis.

Fluorescence microscopy point spread function model accounting for aberrations due to refractive index variability within a specimen.

PubMed

Ghosh, Sreya; Preza, Chrysanthe

2015-07-01

A three-dimensional (3-D) point spread function (PSF) model for wide-field fluorescence microscopy, suitable for imaging samples with variable refractive index (RI) in multilayered media, is presented. This PSF model is a key component for accurate 3-D image restoration of thick biological samples, such as lung tissue. Microscope- and specimen-derived parameters are combined with a rigorous vectorial formulation to obtain a new PSF model that accounts for additional aberrations due to specimen RI variability. Experimental evaluation and verification of the PSF model was accomplished using images from 175-nm fluorescent beads in a controlled test sample. Fundamental experimental validation of the advantage of using improved PSFs in depth-variant restoration was accomplished by restoring experimental data from beads (6  μm in diameter) mounted in a sample with RI variation. In the investigated study, improvement in restoration accuracy in the range of 18 to 35% was observed when PSFs from the proposed model were used over restoration using PSFs from an existing model. The new PSF model was further validated by showing that its prediction compares to an experimental PSF (determined from 175-nm beads located below a thick rat lung slice) with a 42% improved accuracy over the current PSF model prediction.

Evaluation of Hydrodynamic Chromatography Coupled with UV-Visible, Fluorescence and Inductively Coupled Plasma Mass Spectrometry Detectors for Sizing and Quantifying Colloids in Environmental Media

PubMed Central

Philippe, Allan; Schaumann, Gabriele E.

2014-01-01

In this study, we evaluated hydrodynamic chromatography (HDC) coupled with inductively coupled plasma mass spectrometry (ICP-MS) for the analysis of nanoparticles in environmental samples. Using two commercially available columns (Polymer Labs-PDSA type 1 and 2), a set of well characterised calibrants and a new external time marking method, we showed that flow rate and eluent composition have few influence on the size resolution and, therefore, can be adapted to the sample particularity. Monitoring the agglomeration of polystyrene nanoparticles over time succeeded without observable disagglomeration suggesting that even weak agglomerates can be measured using HDC. Simultaneous determination of gold colloid concentration and size using ICP-MS detection was validated for elemental concentrations in the ppb range. HDC-ICP-MS was successfully applied to samples containing a high organic and ionic background. Indeed, online combination of UV-visible, fluorescence and ICP-MS detectors allowed distinguishing between organic molecules and inorganic colloids during the analysis of Ag nanoparticles in synthetic surface waters and TiO2 and ZnO nanoparticles in commercial sunscreens. Taken together, our results demonstrate that HDC-ICP-MS is a flexible, sensitive and reliable method to measure the size and the concentration of inorganic colloids in complex media and suggest that there may be a promising future for the application of HDC in environmental science. Nonetheless the rigorous measurements of agglomerates and of matrices containing natural colloids still need to be studied in detail. PMID:24587393

The detection and assessment of the aneugenic potential of selected oestrogens, progestins and androgens using the in vitro cytokinesis blocked micronucleus assay.

PubMed

Kayani, M A; Parry, James M

2008-03-12

The use of 17-beta-oestradiol, testosterone, progesterone, zearanol, trenbolone acetate and melengesterol acetate in animal feed as growth promoters has been banned in the European Union since 1989. However, the data available on their genotoxicity is limited. To bridge this gap the present study was carried out with the aim of evaluating these hormones for their ability to induce aneuploidy. Aneuploidy has been recently considered sufficiently important to be included in the routine testing of chemicals and radiation. These types of numerical chromosomal aberrations may arise by at least two mechanisms, chromosome loss and non-disjunction. Over the past few years, the cytokinesis blocked micronucleus (CBMN) technique has evolved into a robust assay for the detection of aneuploidy induction. At the present time, it is the only assay which can reliably detect both chromosome loss and non-disjunction when the basic methodology is coupled with appropriate molecular probing techniques such as immunoflourescent labelling of kinetochores and Fluorescence in situ Hybridisation. In this present study, aneuploidy induction by three groups of hormones was studied using CBMN assay coupled with Fluorescence in situ Hybridisation. The results from the present study demonstrate that 17-beta-oestradiol, diethylstilboestrol, progesterone and testosterone are genotoxic and induce aneuploidy by non-disjunctional mechanism, whereas trenbolone is also genotoxic by a clastogenic mechanism. However, melengesterol acetate and zearanol proved to be non-genotoxic in vitro.

Effect of oxidation and extent of oxidation on biologically active PACs in asphalt products.

PubMed

Trumbore, David; Osborn, Linda; Blackburn, Gary; Niebo, Ron; Kriech, Anthony; Maxim, L Daniel

2011-10-01

Recent studies have reported divergent results in rodent cancer assays using fume condensates from a variety of asphalt products. This paper presents results of a study investigating the role of oxidation, or extent of oxidation, on these findings. Five straight run asphalts, made from widely used crude oils, were used as inputs to both production scale and laboratory oxidation units and processed to a range of softening points used in common roofing products. For each of the five asphalts studied, the oxidation reaction significantly decreased measures of polycyclic aromatic compounds (PACs) that have been linked, previously and in analyses included in this study, to tumor induction in rodent bioassays. Mutagenicity index determined by the modified Ames assay was reduced between 41% and 50% from the input asphalt to the final oxidized product. A fluorescence method tuned to a subset of PAC compounds that have been associated with carcinogenic behavior in mouse bioassays was reduced between 39% and 71%. The decrease was largest in the first quarter of the oxidation reaction. These findings indicate that oxidation, by itself, was not a likely factor in the tumor induction seen in the previous studies. Rather, other factors such as the conditions of fume generation and crude source (coupled with possible differences in distillation endpoints) were more likely to have determined the outcomes. Analyses of previously published data, presented in this paper, suggest that the modified Ames and fluorescence assays are valuable screening tools for use in future health-related asphalt research.

Nanoparticle-mediated combination chemotherapy and photodynamic therapy overcomes tumor drug resistance in vitro.

PubMed

Khdair, Ayman; Handa, Hitesh; Mao, Guangzhao; Panyam, Jayanth

2009-02-01

Drug resistance limits the success of many anticancer drugs. Reduced accumulation of the drug at its intracellular site of action because of overexpression of efflux transporters such as P-glycoprotein (P-gp) is a major mechanism of drug resistance. In this study, we investigated whether photodynamic therapy (PDT) using methylene blue, also a P-gp inhibitor, can be used to enhance doxorubicin-induced cytotoxicity in drug-resistant tumor cells. Aerosol OT (AOT)-alginate nanoparticles were used as a carrier for the simultaneous cellular delivery of doxorubicin and methylene blue. Methylene blue was photoactivated using light of 665 nm wavelength. Induction of apoptosis and necrosis following treatment with combination chemotherapy and PDT was investigated in drug-resistant NCI/ADR-RES cells using flow cytometry and fluorescence microscopy. Effect of encapsulation in nanoparticles on the intracellular accumulation of doxorubicin and methylene blue was investigated qualitatively using fluorescence microscopy and was quantitated using HPLC. Encapsulation in AOT-alginate nanoparticles significantly enhanced the cytotoxicity of combination therapy in resistant tumor cells. Nanoparticle-mediated combination therapy resulted in a significant induction of both apoptosis and necrosis. Improvement in cytotoxicity could be correlated with enhanced intracellular and nuclear delivery of the two drugs. Further, nanoparticle-mediated combination therapy resulted in significantly elevated reactive oxygen species (ROS) production compared to single drug treatment. In conclusion, nanoparticle-mediated combination chemotherapy and PDT using doxorubicin and methylene blue was able to overcome resistance mechanisms and resulted in improved cytotoxicity in drug-resistant tumor cells.

Chemically Induced Conditional Rescue of the Reduced Epidermal Fluorescence8 Mutant of Arabidopsis Reveals Rapid Restoration of Growth and Selective Turnover of Secondary Metabolite Pools1[C][OPEN

PubMed Central

Kim, Jeong Im; Ciesielski, Peter N.; Donohoe, Bryon S.; Chapple, Clint; Li, Xu

2014-01-01

The phenylpropanoid pathway is responsible for the biosynthesis of diverse and important secondary metabolites including lignin and flavonoids. The reduced epidermal fluorescence8 (ref8) mutant of Arabidopsis (Arabidopsis thaliana), which is defective in a lignin biosynthetic enzyme p-coumaroyl shikimate 3′-hydroxylase (C3′H), exhibits severe dwarfism and sterility. To better understand the impact of perturbation of phenylpropanoid metabolism on plant growth, we generated a chemically inducible C3′H expression construct and transformed it into the ref8 mutant. Application of dexamethasone to these plants greatly alleviates the dwarfism and sterility and substantially reverses the biochemical phenotypes of ref8 plants, including the reduction of lignin content and hyperaccumulation of flavonoids and p-coumarate esters. Induction of C3′H expression at different developmental stages has distinct impacts on plant growth. Although early induction effectively restored the elongation of primary inflorescence stem, application to 7-week-old plants enabled them to produce new rosette inflorescence stems. Examination of hypocotyls of these plants revealed normal vasculature in the newly formed secondary xylem, presumably restoring water transport in the mutant. The ref8 mutant accumulates higher levels of salicylic acid than the wild type, but depletion of this compound in ref8 did not relieve the mutant’s growth defects, suggesting that the hyperaccumulation of salicylic acid is unlikely to be responsible for dwarfism in this mutant. PMID:24381065

Evaluation of hydrodynamic chromatography coupled with UV-visible, fluorescence and inductively coupled plasma mass spectrometry detectors for sizing and quantifying colloids in environmental media.

PubMed

Philippe, Allan; Schaumann, Gabriele E

2014-01-01

In this study, we evaluated hydrodynamic chromatography (HDC) coupled with inductively coupled plasma mass spectrometry (ICP-MS) for the analysis of nanoparticles in environmental samples. Using two commercially available columns (Polymer Labs-PDSA type 1 and 2), a set of well characterised calibrants and a new external time marking method, we showed that flow rate and eluent composition have few influence on the size resolution and, therefore, can be adapted to the sample particularity. Monitoring the agglomeration of polystyrene nanoparticles over time succeeded without observable disagglomeration suggesting that even weak agglomerates can be measured using HDC. Simultaneous determination of gold colloid concentration and size using ICP-MS detection was validated for elemental concentrations in the ppb range. HDC-ICP-MS was successfully applied to samples containing a high organic and ionic background. Indeed, online combination of UV-visible, fluorescence and ICP-MS detectors allowed distinguishing between organic molecules and inorganic colloids during the analysis of Ag nanoparticles in synthetic surface waters and TiO₂ and ZnO nanoparticles in commercial sunscreens. Taken together, our results demonstrate that HDC-ICP-MS is a flexible, sensitive and reliable method to measure the size and the concentration of inorganic colloids in complex media and suggest that there may be a promising future for the application of HDC in environmental science. Nonetheless the rigorous measurements of agglomerates and of matrices containing natural colloids still need to be studied in detail.

Size of submicrometric and nanometric particles affect cellular uptake and biological activity of macrophages in vitro.

PubMed

Leclerc, L; Rima, W; Boudard, D; Pourchez, J; Forest, V; Bin, V; Mowat, P; Perriat, P; Tillement, O; Grosseau, P; Bernache-Assollant, D; Cottier, M

2012-08-01

Micrometric and nanometric particles are increasingly used in different fields and may exhibit variable toxicity levels depending on their physicochemical characteristics. The aim of this study was to determine the impact of the size parameter on cellular uptake and biological activity, working with well-characterized fluorescent particles. We focused our attention on macrophages, the main target cells of the respiratory system responsible for the phagocytosis of the particles. FITC fluorescent silica particles of variable submicronic sizes (850, 500, 250 and 150 nm) but with similar surface coating (COOH) were tailored and physico-chemically characterized. These particles were then incubated with the RAW 264.7 macrophage cell line. After microscopic observations (SEM, TEM, confocal), a quantitative evaluation of the uptake was carried out. Fluorescence detected after a quenching with trypan blue allows us to distinguish and quantify entirely engulfed fluorescent particles from those just adhering to the cell membrane. Finally, these data were compared to the in vitro toxicity assessed in terms of cell damage, inflammation and oxidative stress (evaluated by LDH release, TNF-α and ROS production respectively). Particles were well characterized (fluorescence, size distribution, zeta potential, agglomeration and surface groups) and easily visualized after cellular uptake using confocal and electron microscopy. The number of internalized particles was precisely evaluated. Size was found to be an important parameter regarding particles uptake and in vitro toxicity but this latter strongly depends on the particles doses employed.

Mitochondrial distribution and activity in human mature oocytes: gonadotropin-releasing hormone agonist versus antagonist for pituitary down-regulation.

PubMed

Dell'Aquila, Maria Elena; Ambruosi, Barbara; De Santis, Teresa; Cho, Yoon Sung

2009-01-01

To analyze the effects of GnRH agonists versus antagonists on mitochondrial distribution and activity in human mature oocytes. Randomized research experimental study. Academic basic research laboratory and hospital-based fertility center. Two hundred twenty-five supernumerary mature oocytes from 44 patients. Fluorescent staining and confocal laser scanning microscopy on oocytes after the use of either GnRH agonist (group A) or GnRH antagonist (group B). Oocyte mitochondrial distribution pattern and activity using MitoTracker Orange CMTM Ros. More oocytes showing polarized mitochondrial distribution pattern were found in group A than in group B (35% vs. 14%). In group B, hCG rather than GnRH agonist, for ovulation induction, resulted in more oocytes showing heterogeneous (57% vs. 14%), in particular polarized (24% vs. 0) mitochondrial distribution. In groups A and B, fluorescence intensity did not vary according to mitochondrial distribution pattern. However, fluorescence intensity was higher in oocytes with polarized and large granules configurations in group B compared to group A. The GnRH agonist and antagonist may have different effects on oocyte mitochondrial distribution pattern and activity. The GnRH antagonist may induce mitochondrial hyperactivity, which may be detrimental to the oocyte.

In vitro translation with [34S]-labeled methionine, selenomethionine, and telluromethionine.

PubMed

Ogra, Yasumitsu; Kitaguchi, Takashi; Suzuki, Noriyuki; Suzuki, Kazuo T

2008-01-01

Heteroisotope and heteroatom tagging with [(34)S]-enriched methionine (Met), selenomethionine (SeMet), and telluromethionine (TeMet) was applied to in vitro translation. Green fluorescent protein (GFP) and JNK stimulatory phosphatase-1 (JSP-1) genes were translated with wheat germ extract (WGE) in the presence of Met derivatives. GFPs containing Met derivatives were subjected to HPLC coupled with treble detection, i.e., a photodiode array detector, a fluorescence detector, and an inductively coupled plasma mass spectrometer (ICP-MS). The activities of JSP-1-containing Met derivatives were also measured. GFP and JSP-1 containing [(34)S]-Met and SeMet showed comparable fluorescence intensities and enzyme activities to those containing naturally occurring Met. TeMet was unstable and decomposed in WGE, whereas SeMet was stable throughout the experimental period. Thus, although Te was the most sensitive to ICP-MS detection among S, Se, and Te, TeMet was less incorporated into the proteins than Met and SeMet. Finally, the potential of heteroisotope and heteroatom tagging of desired proteins in in vitro translation followed by ICP-MS detection was discussed. [figure: see text] TeMet was less incorporated into GFP than Met and SeMet due to its instability in WGE.

Detection of intramyocardially injected DiR-labeled mesenchymal stem cells by optical and optoacoustic tomography.

PubMed

Berninger, Markus T; Mohajerani, Pouyan; Wildgruber, Moritz; Beziere, Nicolas; Kimm, Melanie A; Ma, Xiaopeng; Haller, Bernhard; Fleming, Megan J; Vogt, Stephan; Anton, Martina; Imhoff, Andreas B; Ntziachristos, Vasilis; Meier, Reinhard; Henning, Tobias D

2017-06-01

The distribution of intramyocardially injected rabbit MSCs, labeled with the near-infrared dye 1,1'-dioctadecyl-3,3,3',3'-tetramethylindotricarbo-cyanine-iodide (DiR) using hybrid Fluorescence Molecular Tomography-X-ray Computed Tomography (FMT-XCT) and Multispectral Optoacoustic Tomography (MSOT) imaging technologies, was investigated. Viability and induction of apoptosis of DiR labeled MSCs were assessed by XTT- and Caspase-3/-7-testing in vitro . 2 × 10 6 , 2 × 10 5 and 2 × 10 4 MSCs labeled with 5 and 10 μg DiR/ml were injected into fresh frozen rabbit hearts. FMT-XCT, MSOT and fluorescence cryosection imaging were performed. Concentrations up to 10 μg DiR/ml did not cause apoptosis in vitro (p > 0.05). FMT and MSOT imaging of labeled MSCs led to a strong signal. The imaging modalities highlighted a difference in cell distribution and concentration correlated to the number of injected cells. Ex-vivo cryosectioning confirmed the molecular fluorescence signal. FMT and MSOT are sensitive imaging techniques offering high-anatomic resolution in terms of detection and distribution of intramyocardially injected stem cells in a rabbit model.

NIST/ISAC standardization study: variability in assignment of intensity values to fluorescence standard beads and in cross calibration of standard beads to hard dyed beads.

PubMed

Hoffman, Robert A; Wang, Lili; Bigos, Martin; Nolan, John P

2012-09-01

Results from a standardization study cosponsored by the International Society for Advancement of Cytometry (ISAC) and the US National Institute of Standards and Technology (NIST) are reported. The study evaluated the variability of assigning intensity values to fluorophore standard beads by bead manufacturers and the variability of cross calibrating the standard beads to stained polymer beads (hard-dyed beads) using different flow cytometers. Hard dyed beads are generally not spectrally matched to the fluorophores used to stain cells, and spectral response varies among flow cytometers. Thus if hard dyed beads are used as fluorescence calibrators, one expects calibration for specific fluorophores (e.g., FITC or PE) to vary among different instruments. Using standard beads surface-stained with specific fluorophores (FITC, PE, APC, and Pacific Blue™), the study compared the measured intensity of fluorophore standard beads to that of hard dyed beads through cross calibration on 133 different flow cytometers. Using robust CV as a measure of variability, the variation of cross calibrated values was typically 20% or more for a particular hard dyed bead in a specific detection channel. The variation across different instrument models was often greater than the variation within a particular instrument model. As a separate part of the study, NIST and four bead manufacturers used a NIST supplied protocol and calibrated fluorophore solution standards to assign intensity values to the fluorophore beads. Values assigned to the reference beads by different groups varied by orders of magnitude in most cases, reflecting differences in instrumentation used to perform the calibration. The study concluded that the use of any spectrally unmatched hard dyed bead as a general fluorescence calibrator must be verified and characterized for every particular instrument model. Close interaction between bead manufacturers and NIST is recommended to have reliable and uniformly assigned fluorescence standard beads. Copyright © 2012 International Society for Advancement of Cytometry.

A pulse-forming network for particle path visualization. [at Ames Aeromechanics Water Tunnel Facility

NASA Technical Reports Server (NTRS)

Mcalister, K. W.

1981-01-01

A procedure is described for visualizing nonsteady fluid flow patterns over a wide velocity range using discrete nonluminous particles. The paramount element responsible for this capability is a pulse-forming network with variable inductance that is used to modulate the discharge of a fixed amount of electrical energy through a xenon flashtube. The selectable duration of the resultant light emission functions as a variable shutter so that particle path images of constant length can be recorded. The particles employed as flow markers are hydrogen bubbles that are generated by electrolysis in a water tunnel. Data are presented which document the characteristics of the electrical circuit and establish the relation of particle velocity to both section inductance and film exposure.

Cultural Background Variables in Dance Talent Development: Findings from the UK Centres for Advanced Training

ERIC Educational Resources Information Center

Sanchez, Erin N.; Aujla, Imogen J.; Nordin-Bates, Sanna

2013-01-01

This study is a qualitative enquiry into cultural background variables--social support, values, race/ethnicity and economic means--in the process of dance talent development. Seven urban dance students in pre-vocational training, aged 15-19, participated in semi-structured interviews. Interviews were inductively analysed using QSR International…

Rational Variability in Children's Causal Inferences: The Sampling Hypothesis

ERIC Educational Resources Information Center

Denison, Stephanie; Bonawitz, Elizabeth; Gopnik, Alison; Griffiths, Thomas L.

2013-01-01

We present a proposal--"The Sampling Hypothesis"--suggesting that the variability in young children's responses may be part of a rational strategy for inductive inference. In particular, we argue that young learners may be randomly sampling from the set of possible hypotheses that explain the observed data, producing different hypotheses with…

Highways and Byways: The Career Paths of Senior Student Affairs Officers

ERIC Educational Resources Information Center

Tull, Ashley; Miller, Michael T.

2009-01-01

The highways and byways, or career paths, to the Senior Student Affairs Officer (SSAO) position differ based on a variety of variables. This study examined several variables including the induction or graduate preparation process, professional pathways, and professional and academic involvement of more than half of current land grant SSAOs. Data…

A randomized trial of misoprostol versus extra-amniotic sodium chloride infusion with oxytocin for induction of labor.

PubMed

Buccellato, C A; Stika, C S; Frederiksen, M C

2000-05-01

Our purpose was to compare the efficacy and safety of misoprostol and extra-amniotic sodium chloride infusion with oxytocin for induction of labor. This randomized trial compared two methods of labor induction in women requiring cervical ripening. One hundred twenty-three women undergoing labor induction with a Bishop score < or =5 were randomly selected to receive either misoprostol, 50 microg intravaginally every 4 hours, or extra-amniotic sodium chloride infusion. The primary outcome variable was the time interval from induction to vaginal delivery. Sixty-one women received extra-amniotic sodium chloride infusion and 62 women received misoprostol. The mean time interval from the start of induction to vaginal delivery was 15.0 +/- 5.0 hours and 16.5 +/- 7.2 hours for the extra-amniotic infusion and misoprostol groups, respectively (P, not significant). The cesarean delivery rate was not significantly different between the 2 groups (32.8% for the extra-amniotic infusion group; 19.4% for the misoprostol group). Maternal and neonatal outcomes were similar between the 2 groups. Both methods of induction are equally efficacious and result in similar maternal and neonatal outcomes.

Mood state dependency of dysfunctional attitudes in bipolar affective disorder.

PubMed

Babakhani, Anet; Startup, Mike

2012-01-01

Studies of cognitive styles among euthymic people with bipolar affective disorder (BAD) without use of mood induction techniques to access those cognitive styles give misleading impressions of normality of those cognitions. The aim of this study was to assess dysfunctional attitudes of participants with BAD, and control participants with no previous psychiatric histories, after mood inductions. Sad and happy moods were induced within 49 BAD and 37 controls. Dysfunctional attitudes were measured following mood inductions using the Dysfunctional Attitude Scale-short form (DAS-24), which has three subscales of achievement, interpersonal, and goal attainment. It was hypothesised that within BAD the sad mood induction would help in accessing dysfunctional attitudes in all three domains relative to the happy mood induction. This was supported. It was also hypothesised that the mood inductions would not affect dysfunctional attitudes within controls. This was supported. When diagnosis was entered as a between group variable, achievement dysfunctional attitudes were significantly higher in BAD compared to controls after a happy induction. Both sad and happy moods provoked higher levels of dysfunctional attitudes within BAD. Euphoria may be related to elevated achievement dysfunctional attitudes, raising risk for mania.

Transvaginal ultrasonographic cervical measurement in predicting failed labor induction and cesarean delivery for failure to progress in nulliparous women.

PubMed

Park, Kyo Hoon

2007-08-01

The aim of this study was to evaluate the value of transvaginal sonographic cervical measurement in predicting failed labor induction and cesarean delivery for failure to progress in nulliparous women. One hundred and sixty-one women scheduled for labor induction underwent transvaginal ultrasonography and digital cervical examinations. Logistic regression demonstrated that cervical length and gestational age at induction, but not the Bishop score, significantly and independently predicted failed labor induction. According to the receiver operating characteristic curves analysis, the best cut-off value of cervical length for predicting failed labor induction was 28 mm, with a sensitivity of 62% and a specificity of 60%. In terms of the likelihood of a cesarean delivery for failure to progress as the outcome variable, logistic regression indicated that maternal height and birth weight, but not cervical length or Bishop score, were significantly and independently associated with an increased risk of cesarean delivery for failure to progress. Transvaginal sonographic measurements of cervical length thus independently predicted failed labor induction in nulliparous women. However, the relatively poor predictive performance of this test undermines its clinical usefulness as a predictor of failed labor induction. Moreover, cervical length appears to have a poor predictive value for the likelihood of a cesarean delivery for failure to progress.

On Variability in Satellite Terrestrial Chlorophyll Fluorescence Measurements: Relationships with Phenology and Ecosystem-Atmosphere Carbon Exchange, Vegetation Structure, Clouds, and Sun-Satellite Geometry

NASA Astrophysics Data System (ADS)

Joiner, J.; Yoshida, Y.; Guanter, L.; Zhang, Y.; Vasilkov, A. P.; Schaefer, K. M.; Huemmrich, K. F.; Middleton, E.; Koehler, P.; Jung, M.; Tucker, C. J.; Lyapustin, A.; Wang, Y.; Frankenberg, C.; Berry, J. A.; Koster, R. D.; Reichle, R. H.; Lee, J. E.; Kawa, S. R.; Collatz, G. J.; Walker, G. K.; Van der Tol, C.

2014-12-01

Over the past several years, there have been several breakthroughs in our ability to detect the very small fluorescence emitted by chlorophyll in vegetation globally from space. There are now multiple instruments in space capable of measuring this signal at varying temporal and spatial resolutions. We will review the state-of-the-art with respect to these relatively new satellite measurements and ongoing studies that examine the relationships with photosynthesis. Now that we have a data record spanning more than seven years, we can examine variations due to seasonal carbon uptake, interannual variability, land-use changes, and water and temperature stress. In addition, we examine how clouds and satellite viewing geometry impact the signal. We compare and contrast these variations with those from popular vegetation indices, such as the Normalized Difference Vegetation Index (NDVI), related to the potential photosynthesis as well as with measurements from flux tower gas exchange measurements and other model-based estimates of Global Primary Productivity (GPP). Vegetation fluorescence can be simulated in global vegetation models as well as with 1D canopy radiative transport models. We will describe how the satellite fluorescence data are being used to evaluate and potentially improve these models.

Vitellogenin synthesis in primary cultures of fish liver cells as endpoint for in vitro screening of the (anti)estrogenic activity of chemical substances.

PubMed

Navas, José M; Segner, Helmut

2006-10-25

Concern over possible adverse effects of endocrine-disrupting compounds on fish has caused the development of appropriate testing methods. In vitro screening assays may provide initial information on endocrine activities of a test compound and thereby may direct and optimize subsequent testing. Induction of vitellogenin (VTG) is used as a biomarker of exposure of fish to estrogen-active substances. Since VTG induction can be measured not only in vivo but also in fish hepatocytes in vitro, the use of VTG induction response in isolated fish liver cells has been suggested as in vitro screen for identifying estrogenic-active substances. The main advantages of the hepatocyte VTG assay are considered its ability to detect effects of estrogenic metabolites, since hepatocytes in vitro remain metabolically competent, and its ability to detect both estrogenic and anti-estrogenic effects. In this article, we critically review the current knowledge on the VTG response of cultured fish hepatocytes to (anti)estrogenic substances. In particular, we discuss the sensitivity, specificity, and variability of the VTG hepatocyte assay. In addition, we review the available data on culture factors influencing basal and induced VTG production, the response to natural and synthetic estrogens as well as to xenoestrogens, the detection of indirect estrogens, and the sources of assay variability. The VTG induction in cultured fish hepatocytes is clearly influenced by culture conditions (medium composition, temperature, etc.) and culture system (hepatocyte monolayers, aggregates, liver slices, etc.). The currently available database on estrogen-mediated VTG induction in cultured teleost hepatocytes is too small to support conclusive statements on whether there exist systematic differences of the VTG response between in vitro culture systems, VTG analytical methods or fish species. The VTG hepatocyte assay detects sensitively natural and synthetic estrogens, whereas the response to xenoestrogens appears to be more variable. The detection of weak estrogens can be critical due to the overshadow with cytotoxic concentrations. Moreover, the VTG hepatocyte assay is able to detect antiestrogens as well as indirect estrogens, i.e substances which require metabolic activation to induce an estrogenic response. Nevertheless, more chemicals need to be analysed to corroborate this statement. It will be necessary to establish standardized protocols to minimize assay variability, and to develop a set of pass-fail criteria as well as cut-offs for designating positive and negative responses.

Behavior of Excited Argon Atoms in Inductively Driven Plasmas

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

HEBNER,GREGORY A.; MILLER,PAUL A.

1999-12-07

Laser induced fluorescence has been used to measure the spatial distribution of the two lowest energy argon excited states, 1s{sub 5} and 1s{sub 4}, in inductively driven plasmas containing argon, chlorine and boron trichloride. The behavior of the two energy levels with plasma conditions was significantly different, probably because the 1s{sub 5} level is metastable and the 1s{sub 4} level is radiatively coupled to the ground state but is radiation trapped. The argon data is compared with a global model to identify the relative importance of processes such as electron collisional mixing and radiation trapping. The trends in the datamore » suggest that both processes play a major role in determining the excited state density. At lower rfpower and pressure, excited state spatial distributions in pure argon were peaked in the center of the discharge, with an approximately Gaussian profile. However, for the highest rfpowers and pressures investigated, the spatial distributions tended to flatten in the center of the discharge while the density at the edge of the discharge was unaffected. The spatially resolved excited state density measurements were combined with previous line integrated measurements in the same discharge geometry to derive spatially resolved, absolute densities of the 1s{sub 5} and 1s{sub 4} argon excited states and gas temperature spatial distributions. Fluorescence lifetime was a strong fi.mction of the rf power, pressure, argon fraction and spatial location. Increasing the power or pressure resulted in a factor of two decrease in the fluorescence lifetime while adding Cl{sub 2} or BCl{sub 3} increased the fluorescence lifetime. Excited state quenching rates are derived from the data. When Cl{sub 2} or BCl{sub 3} was added to the plasma, the maximum argon metastable density depended on the gas and ratio. When chlorine was added to the argon plasma, the spatial density profiles were independent of chlorine fraction. While it is energetically possible for argon excited states to dissociate some of the molecular species present in this discharge, it does not appear to be a significant source of dissociation. The major source of interaction between the argon and the molecular species BCl{sub 3} and Cl{sub 2} appears to be through modification of the electron density.« less

Changes in heart rate variability during the induction and decay of heat acclimation.

PubMed

Flouris, Andreas D; Poirier, Martin P; Bravi, Andrea; Wright-Beatty, Heather E; Herry, Christophe; Seely, Andrew J; Kenny, Glen P

2014-10-01

We evaluated the changes in core temperature, heart rate, and heart rate variability (HRV) during the induction and decay of heat acclimation. Ten males (23 ± 3 years; 79.5 ± 3.5 kg; 15.2 ± 4.5 percent body fat; 51.13 ± 4.61 mLO(2)∙kg(-1)∙min(-1) peak oxygen uptake) underwent a 14-day heat acclimation protocol comprising of 90-min cycling at ~50 % peak oxygen uptake at 40 °C and ~20 % relative humidity. Core temperature, heart rate, and 102 HRV measures were recorded during a heat tolerance test conducted at baseline (day 0) and at the end of the induction (day 14) and decay (day 28) phases. Heat acclimation resulted in significantly reduced core temperature [rectal (χ (2) = 1298.14, p < 0.001); esophageal (χ (2) = 1069.88, p < 0.001)] and heart rate (χ (2) = 1230.17, p < 0.001). Following the decay phase, 26, 40, and 60 % of the heat acclimation-induced reductions in rectal temperature, esophageal temperature, and heart rate, respectively, were lost. Heat acclimation was accompanied by profound and broad changes in HRV: at the end of the induction phase, 75 of the 102 variability measures computed were significantly different (p < 0.001), compared to only 47 of the 102 at the end of the decay phase. Heat acclimation is accompanied by reduced core temperature, significant bradycardia, and marked alterations in HRV, which we interpret as being related to vagal dominance. The observed changes in core temperature persist for at least 2 weeks of non-exposure to heat, while the changes in heart rate and HRV decay faster and are only partly evident after 2 weeks of non-exposure to heat.

Dose requirements of alfentanil to eliminate autonomic responses during rapid-sequence induction with thiopental 4 mg/kg and rocuronium 0.6 mg/kg.

PubMed

Abou-Arab, Mohammad H; Rostrup, Morten; Heier, Tom

2016-12-01

Opioids are integral part of anesthesia induction, but information on optimal dosing is limited. We aimed to determine doses of alfentanil needed to eliminate increases in 5 autonomic response variables (plasma concentrations of epinephrine, norepinephrine and vasopressin, arterial blood pressure [ABP], and heart rate) during rapid-sequence induction of anesthesia with thiopental 4 mg/kg and rocuronium 0.6 mg/kg. Prospective, randomized, observer-blinded, interventional clinical study. Large academic institution. Eighty-four healthy patients, aged 18 to 55 years, received 1 of 7 assessor-blinded doses of alfentanil (0, 10, 20, 30, 40, 50, and 60 μg/kg) together with thiopental 4 mg/kg and rocuronium 0.6 mg/kg, administered in rapid succession (15 seconds). Laryngoscopy was initiated 40 seconds after rocuronium, and tracheal intubation was concluded within 15 seconds thereafter. An indwelling radial artery catheter was used for hemodynamic monitoring and blood sampling. Relationships between alfentanil dose and response variables were tested with linear regression, and the influence of covariates (sex, body weight, and age) was determined. Alfentanil dose needed to prevent increases in ABP >10% above baseline with 95% probability was estimated with logistic regression. Significant relationships were determined between alfentanil dose and response variables. Clinically interesting influence of covariates was not found. Alfentanil 55 μg/kg was needed to prevent increases in ABP postintubation >10% above baseline with 95% probability. One individual needed a bolus of vasopressor postintubation. Optimal control of autonomic responses during rapid-sequence induction was achieved with clinically relevant doses of alfentanil in healthy patients anesthetized with thiopental 4 mg/kg and rocuronium 0.6 mg/kg. Copyright © 2016 Elsevier Inc. All rights reserved.

Baseline Chromatin Modification Levels May Predict ...

EPA Pesticide Factsheets

Traditional toxicological paradigms have relied on factors such as age, genotype, and disease status to explain variability in responsiveness to toxicant exposure; however, these are neither sufficient to faithfully identify differentially responsive individuals nor are they modifiable factors that can be leveraged to mitigate the exposure effects. Unlike these factors, the epigenome is dynamic and shaped by an individual's environment. We sought to determine whether baseline levels of specific chromatin modifications correlated with the interindividual variability in their ozone (03)-mediated induction in an air-liquid interface model using primary human bronchial epithelial cells from a panel of 11 donors. We characterized the relationship between the baseline abundance of 6 epigenetic markers with established roles as key regulators of gene expression-histone H3 lysine 4 trimethylation (H3K4me3), H3K27 acetylation (H3K27ac), pan­acetyl H4 (H4ac), histone H3K27 di/trimethylation (H3K27me2/3), unmodified H3, and5-hydroxymethylcytosine (5-hmC)-and the variability in the 03-induced expression of IL-8, IL-6, COX2, and HMOX1. Baseline levels of H3K4me3, H3K27me2/3, and 5-hmC, but not H3K27ac, H4ac, and total H3, correlated with the interindividual variability in 03-mediated induction of HMOX1 and COX2. In contrast, none of the chromatin modifications that we examined correlated with the induction of IL-8 and IL-6. From these findings, we propose an "epigenetic see

Counterfeit-resistant materials and a method and apparatus for authenticating materials

DOEpatents

Ramsey, J. Michael; Klatt, Leon N.

2001-01-01

Fluorescent dichroic fibers randomly incorporated within a media provide an improved method for authentication and counterfeiting protection. The dichroism is provided by an alignment of fluorescent molecules along the length of the fibers. The fluorescent fibers provide an authentication mechanism of varying levels of capability. The authentication signature depends on four parameters, the x,y position, the dichroism and the local environment. The availability of so many non-deterministic variables makes production of counterfeit articles (e.g., currency, credit cards, etc.) essentially impossible Counterfeit-resistant articles, an apparatus for authenticating articles, and a process for forming counterfeit-resistant media are also provided&

Counterfeit-resistant materials and a method and apparatus for authenticating materials

DOEpatents

Ramsey, J. Michael; Klatt, Leon N.

2000-01-01

Fluorescent dichroic fibers randomly incorporated within a media provide an improved method for authentication and counterfeiting protection. The dichroism is provided by an alignment of fluorescent molecules along the length of the fibers. The fluorescent fibers provide an authentication mechanism of varying levels of capability. The authentication signature depends on four parameters; the x,y position, the dichroism and the local environment. The availability of so many non-deterministic variables makes production of counterfeit articles (e.g., currency, credit cards, etc.) essentially impossible. Counterfeit-resistant articles, an apparatus for authenticating articles, and a process for forming counterfeit-resistant media are also provided.

Design and testing of a coil-unit barrel for helical coil electromagnetic launcher

NASA Astrophysics Data System (ADS)

Yang, Dong; Liu, Zhenxiang; Shu, Ting; Yang, Lijia; Ouyang, Jianming

2018-01-01

A coil-unit barrel for a helical coil electromagnetic launcher is described. It provides better features of high structural strength and flexible adjustability. It is convenient to replace the damaged coil units and easy to adjust the number of turns in the stator coils due to the modular design. In our experiments, the highest velocity measured for a 4.5-kg projectile is 47.3 m/s and the mechanical reinforcement of the launcher could bear 35 kA peak current. The relationship between the energy conversion efficiency and the inductance gradient of the launcher is also studied. In the region of low inductance gradient, the efficiency is positively correlated with the inductance gradient. However, in the region of high inductance gradient, the inter-turn arc erosion becomes a major problem of limiting the efficiency and velocity of the launcher. This modular barrel allows further studies in the inter-turn arc and the variable inductance gradient helical coil launcher.

Design and testing of a coil-unit barrel for helical coil electromagnetic launcher.

PubMed

Yang, Dong; Liu, Zhenxiang; Shu, Ting; Yang, Lijia; Ouyang, Jianming

2018-01-01

A coil-unit barrel for a helical coil electromagnetic launcher is described. It provides better features of high structural strength and flexible adjustability. It is convenient to replace the damaged coil units and easy to adjust the number of turns in the stator coils due to the modular design. In our experiments, the highest velocity measured for a 4.5-kg projectile is 47.3 m/s and the mechanical reinforcement of the launcher could bear 35 kA peak current. The relationship between the energy conversion efficiency and the inductance gradient of the launcher is also studied. In the region of low inductance gradient, the efficiency is positively correlated with the inductance gradient. However, in the region of high inductance gradient, the inter-turn arc erosion becomes a major problem of limiting the efficiency and velocity of the launcher. This modular barrel allows further studies in the inter-turn arc and the variable inductance gradient helical coil launcher.

Discrimination of corn from monocotyledonous weeds with ultraviolet (UV) induced fluorescence.

PubMed

Panneton, Bernard; Guillaume, Serge; Samson, Guy; Roger, Jean-Michel

2011-01-01

In production agriculture, savings in herbicides can be achieved if weeds can be discriminated from crop, allowing the targeting of weed control to weed-infested areas only. Previous studies demonstrated the potential of ultraviolet (UV) induced fluorescence to discriminate corn from weeds and recently, robust models have been obtained for the discrimination between monocots (including corn) and dicots. Here, we developed a new approach to achieve robust discrimination of monocot weeds from corn. To this end, four corn hybrids (Elite 60T05, Monsanto DKC 26-78, Pioneer 39Y85 (RR), and Syngenta N2555 (Bt, LL)) and four monocot weeds (Digitaria ischaemum (Schreb.) I, Echinochloa crus-galli (L.) Beauv., Panicum capillare (L.), and Setaria glauca (L.) Beauv.) were grown either in a greenhouse or in a growth cabinet and UV (327 nm) induced fluorescence spectra (400 to 755 nm) were measured under controlled or uncontrolled ambient light intensity and temperature. This resulted in three contrasting data sets suitable for testing the robustness of discrimination models. In the blue-green region (400 to 550 nm), the shape of the spectra did not contain any useful information for discrimination. Therefore, the integral of the blue-green region (415 to 455 nm) was used as a normalizing factor for the red fluorescence intensity (670 to 755 nm). The shape of the normalized red fluorescence spectra did not contribute to the discrimination and in the end, only the integral of the normalized red fluorescence intensity was left as a single discriminant variable. Applying a threshold on this variable minimizing the classification error resulted in calibration errors ranging from 14.2% to 15.8%, but this threshold varied largely between data sets. Therefore, to achieve robustness, a model calibration scheme was developed based on the collection of a calibration data set from 75 corn plants. From this set, a new threshold can be estimated as the 85% quantile on the cumulative frequency curve of the integral of the normalized red fluorescence. With this approach the classification error was nearly constant (16.0% to 18.5%), thereby indicating the potential of UV-induced fluorescence to reliably discriminate corn from monocot weeds.

Rotational multispectral fluorescence lifetime imaging and intravascular ultrasound: bimodal system for intravascular applications

PubMed Central

Ma, Dinglong; Bec, Julien; Yankelevich, Diego R.; Gorpas, Dimitris; Fatakdawala, Hussain; Marcu, Laura

2014-01-01

Abstract. We report the development and validation of a hybrid intravascular diagnostic system combining multispectral fluorescence lifetime imaging (FLIm) and intravascular ultrasound (IVUS) for cardiovascular imaging applications. A prototype FLIm system based on fluorescence pulse sampling technique providing information on artery biochemical composition was integrated with a commercial IVUS system providing information on artery morphology. A customized 3-Fr bimodal catheter combining a rotational side-view fiberoptic and a 40-MHz IVUS transducer was constructed for sequential helical scanning (rotation and pullback) of tubular structures. Validation of this bimodal approach was conducted in pig heart coronary arteries. Spatial resolution, fluorescence detection efficiency, pulse broadening effect, and lifetime measurement variability of the FLIm system were systematically evaluated. Current results show that this system is capable of temporarily resolving the fluorescence emission simultaneously in multiple spectral channels in a single pullback sequence. Accurate measurements of fluorescence decay characteristics from arterial segments can be obtained rapidly (e.g., 20 mm in 5 s), and accurate co-registration of fluorescence and ultrasound features can be achieved. The current finding demonstrates the compatibility of FLIm instrumentation with in vivo clinical investigations and its potential to complement conventional IVUS during catheterization procedures. PMID:24898604

Mutation induction in bacteria after heavy ion irradiation

NASA Technical Reports Server (NTRS)

Horneck, G.; Kozubek, S.

1994-01-01

From a compilation of experimental data on the mutagenic effects of heavy ions in bacteria, main conclusions have been drawn as follows: (1) The mutagenic efficacy of heavy ions in bacteria depends on physical and biological variables. Physical variables are the radiation dose, energy and charge of the ion; the biological variables are the bacterial strain, the repair genotype of bacteria, and the endpoint investigated (type of mutation, induction of enzymes related to mutagenesis); (2) The responses on dose or fluence are mainly linear or linear quadratic. The quadratic component, if found for low LET radiation, is gradually reduced with increasing LET; (3) At low values of Z and LET the cross section of mutation induction sigma m (as well as SOS response, sigma sos. and lambda phage induction, sigma lambda versus LET curves can be quite consistently described by a common function which increases up to approximately 100 keV/mu m. For higher LET values, the sigma(m) versus LET curves show the so-called 'hooks' observed also for other endpoints; (4) For light ions (Z is less than or equal to 4), the cross sections mostly decrease with increasing ion energy, which is probably related to the decrease of the specific energy departed by the ion inside the sensitive volume (cell). For ions in the range of Z = 10, sigma(m) is nearly independent on the ion energy. For heavier ions (Z is greater than or equal to 16), sigma(m) increases with the energy up to a maximum or saturation around 10 MeV/u. The increment becomes steeper with increasing atomic number of the ion. It correlates with the increasing track radius of the heavy ion; (5) The mutagenic efficiency per lethal event changes slightly with ion energy, if Z is small indicating a rough correlation between cellular lethality and mutation induction, only. For ions of higher Z this relation increases with energy, indicating a change in the 'mode' of radiation action from 'killing-prone' to 'mutation-prone'; and (6) Repair genotype substantially influences the radiation induced mutagenesis. Different mechanisms of mutation induction and/or different types of biologically significant lesions in wild type cells compared to repair deficient strains are a likely explanation.

Elimination of autofluorescence background from fluorescence tissue images by use of time-gated detection and the AzaDiOxaTriAngulenium (ADOTA) fluorophore.

PubMed

Rich, Ryan M; Stankowska, Dorota L; Maliwal, Badri P; Sørensen, Thomas Just; Laursen, Bo W; Krishnamoorthy, Raghu R; Gryczynski, Zygmunt; Borejdo, Julian; Gryczynski, Ignacy; Fudala, Rafal

2013-02-01

Sample autofluorescence (fluorescence of inherent components of tissue and fixative-induced fluorescence) is a significant problem in direct imaging of molecular processes in biological samples. A large variety of naturally occurring fluorescent components in tissue results in broad emission that overlaps the emission of typical fluorescent dyes used for tissue labeling. In addition, autofluorescence is characterized by complex fluorescence intensity decay composed of multiple components whose lifetimes range from sub-nanoseconds to a few nanoseconds. For these reasons, the real fluorescence signal of the probe is difficult to separate from the unwanted autofluorescence. Here we present a method for reducing the autofluorescence problem by utilizing an azadioxatriangulenium (ADOTA) dye with a fluorescence lifetime of approximately 15 ns, much longer than those of most of the components of autofluorescence. A probe with such a long lifetime enables us to use time-gated intensity imaging to separate the signal of the targeting dye from the autofluorescence. We have shown experimentally that by discarding photons detected within the first 20 ns of the excitation pulse, the signal-to-background ratio is improved fivefold. This time-gating eliminates over 96 % of autofluorescence. Analysis using a variable time-gate may enable quantitative determination of the bound probe without the contributions from the background.

Ethidium bromide as a marker of mtDNA replication in living cells

NASA Astrophysics Data System (ADS)

Villa, Anna Maria; Fusi, Paola; Pastori, Valentina; Amicarelli, Giulia; Pozzi, Chiara; Adlerstein, Daniel; Doglia, Silvia Maria

2012-04-01

Mitochondrial DNA (mtDNA) in tumor cells was found to play an important role in maintaining the malignant phenotype. Using laser scanning confocal fluorescence microscopy (LSCFM) in a recent work, we reported a variable fluorescence intensity of ethidium bromide (EB) in mitochondria nucleoids of living carcinoma cells. Since when EB is bound to nucleic acids its fluorescence is intensified; a higher EB fluorescence intensity could reflect a higher DNA accessibility to EB, suggesting a higher mtDNA replication activity. To prove this hypothesis, in the present work we studied, by LSCFM, the EB fluorescence in mitochondria nucleoids of living neuroblastoma cells, a model system in which differentiation affects the level of mtDNA replication. A drastic decrease of fluorescence was observed after differentiation. To correlate EB fluorescence intensity to the mtDNA replication state, we evaluated the mtDNA nascent strands content by ligation-mediated real-time PCR, and we found a halved amount of replicating mtDNA molecules in differentiating cells. A similar result was obtained by BrdU incorporation. These results indicate that the low EB fluorescence of nucleoids in differentiated cells is correlated to a low content of replicating mtDNA, suggesting that EB may be used as a marker of mtDNA replication in living cells.

In vitro Models of Laser Induced Injury: Pathophysiology and Cytoprotection

DTIC Science & Technology

2007-10-29

range of 40-47’C (Dewey, 1979, 1984; Rice, Gray , Dean, & Dewey, 1984; Rice, Gray , & Dewey, 1984; Sapareto & Dewey, 1984). However, most of these in vitro...treated with 1.00 gtg /ml HA for 1 hour, 4 hours, 8 hours, and 24 hours and were then evaluated over a 24-hour time period for production of hsps (Dinh et...Research, 134(3), 331-336. Rice, G. C., Gray , J. W., Dean, P. N., & Dewey, W. C. (1984). Fluorescence-activated cell sorting analysis of the induction

Determination of stoichiometry and concentration of trace elements in thin BaxSr1-xTiO3 perovskite layers.

PubMed

Becker, J S; Boulyga, S F

2001-07-01

This paper describes an analytical procedure for determining the stoichiometry of BaxSr1-xTiO3 perovskite layers using inductively coupled plasma mass spectrometry (ICP-MS). The analytical results of mass spectrometry measurements are compared to those of X-ray fluorescence analysis (XRF). The performance and the limits of solid-state mass spectrometry analytical methods for the surface analysis of thin BaxSr1-xTiO3 perovskite layers sputtered neutral mass spectrometry (SNMS)--are investigated and discussed.

In vitro fluorescence measurements and Monte Carlo simulation of laser irradiation propagation in porcine skin tissue.

PubMed

Drakaki, E; Makropoulou, M; Serafetinides, A A

2008-07-01

In dermatology, the in vivo spectral fluorescence measurements of human skin can serve as a valuable supplement to standard non-invasive techniques for diagnosing various skin diseases. However, quantitative analysis of the fluorescence spectra is complicated by the fact that skin is a complex multi-layered and inhomogeneous organ, with varied optical properties and biophysical characteristics. In this work, we recorded, in vitro, the laser-induced fluorescence emission signals of healthy porcine skin, one of the animals, which is considered as one of the most common models for investigations related to medical diagnostics of human cutaneous tissues. Differences were observed in the form and intensity of the fluorescence signal of the porcine skin, which can be attributed to the different concentrations of the native fluorophores and the variable physical and biological conditions of the skin tissue. As the light transport in the tissue target is directly influencing the absorption and the fluorescence emission signals, we performed Monte Carlo simulation of the light distribution in a five-layer model of human skin tissue, with a pulsed ultraviolet laser beam.

Quantification of cellular autofluorescence of human skin using multiphoton tomography and fluorescence lifetime imaging in two spectral detection channels

PubMed Central

Patalay, Rakesh; Talbot, Clifford; Alexandrov, Yuriy; Munro, Ian; Neil, Mark A. A.; König, Karsten; French, Paul M. W.; Chu, Anthony; Stamp, Gordon W.; Dunsby, Chris

2011-01-01

We explore the diagnostic potential of imaging endogenous fluorophores using two photon microscopy and fluorescence lifetime imaging (FLIM) in human skin with two spectral detection channels. Freshly excised benign dysplastic nevi (DN) and malignant nodular Basal Cell Carcinomas (nBCCs) were excited at 760 nm. The resulting fluorescence signal was binned manually on a cell by cell basis. This improved the reliability of fitting using a double exponential decay model and allowed the fluorescence signatures from different cell populations within the tissue to be identified and studied. We also performed a direct comparison between different diagnostic groups. A statistically significant difference between the median mean fluorescence lifetime of 2.79 ns versus 2.52 ns (blue channel, 300-500 nm) and 2.08 ns versus 1.33 ns (green channel, 500-640 nm) was found between nBCCs and DN respectively, using the Mann-Whitney U test (p < 0.01). Further differences in the distribution of fluorescence lifetime parameters and inter-patient variability are also discussed. PMID:22162820

Developing a scalable model of recombinant protein yield from Pichia pastoris: the influence of culture conditions, biomass and induction regime

PubMed Central

Holmes, William J; Darby, Richard AJ; Wilks, Martin DB; Smith, Rodney; Bill, Roslyn M

2009-01-01

Background The optimisation and scale-up of process conditions leading to high yields of recombinant proteins is an enduring bottleneck in the post-genomic sciences. Typical experiments rely on varying selected parameters through repeated rounds of trial-and-error optimisation. To rationalise this, several groups have recently adopted the 'design of experiments' (DoE) approach frequently used in industry. Studies have focused on parameters such as medium composition, nutrient feed rates and induction of expression in shake flasks or bioreactors, as well as oxygen transfer rates in micro-well plates. In this study we wanted to generate a predictive model that described small-scale screens and to test its scalability to bioreactors. Results Here we demonstrate how the use of a DoE approach in a multi-well mini-bioreactor permitted the rapid establishment of high yielding production phase conditions that could be transferred to a 7 L bioreactor. Using green fluorescent protein secreted from Pichia pastoris, we derived a predictive model of protein yield as a function of the three most commonly-varied process parameters: temperature, pH and the percentage of dissolved oxygen in the culture medium. Importantly, when yield was normalised to culture volume and density, the model was scalable from mL to L working volumes. By increasing pre-induction biomass accumulation, model-predicted yields were further improved. Yield improvement was most significant, however, on varying the fed-batch induction regime to minimise methanol accumulation so that the productivity of the culture increased throughout the whole induction period. These findings suggest the importance of matching the rate of protein production with the host metabolism. Conclusion We demonstrate how a rational, stepwise approach to recombinant protein production screens can reduce process development time. PMID:19570229

Induction of CYP1A1 and CYP1B1 by benzo(k)fluoranthene and benzo(a)pyrene in T-47D human breast cancer cells: Roles of PAH interactions and PAH metabolites

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

Spink, David C.; Wu, Susan J.; Spink, Barbara C.

2008-02-01

The interactions of polycyclic aromatic hydrocarbons (PAH) and cytochromes P450 (CYP) are complex; PAHs are enzyme inducers, substrates, and inhibitors. In T-47D breast cancer cells, exposure to 0.1 to 1 {mu}M benzo(k)fluoranthene (BKF) induced CYP1A1/1B1-catalyzed 17{beta}-estradiol (E{sub 2}) metabolism, whereas BKF levels greater than 1 {mu}M inhibited E{sub 2} metabolism. Time course studies showed that induction of CYP1-catalyzed E{sub 2} metabolism persisted after the disappearance of BKF or co-exposed benzo(a)pyrene, suggesting that BKF metabolites retaining Ah receptor agonist activity were responsible for prolonged CYP1 induction. BKF metabolites were shown, through the use of ethoxyresorufin O-deethylase and CYP1A1-promoter-luciferase reporter assays tomore » induce CYP1A1/1B1 in T-47D cells. Metabolites formed by oxidation at the C-2/C-3 region of BKF had potencies for CYP1 induction exceeding those of BKF, whereas C-8/C-9 oxidative metabolites were somewhat less potent than BKF. The activities of expressed human CYP1A1 and 1B1 with BKF as substrate were investigated by use of HPLC with fluorescence detection, and by GC/MS. The results showed that both enzymes efficiently catalyzed the formation of 3-, 8-, and 9-OHBKF from BKF. These studies indicate that the inductive effects of PAH metabolites as potent CYP1 inducers are likely to be additional important factors in PAH-CYP interactions that affect metabolism and bioactivation of other PAHs, ultimately modulating PAH toxicity and carcinogenicity.« less

Multivariable analysis of anesthetic factors associated with time to extubation in dogs.

PubMed

Kleine, Stephanie; Hofmeister, Erik; Egan, Katrina

2014-12-01

The purpose of this study was to identify factors that prolong the time to extubation in dogs. Anesthetic records of 900 dogs at a university teaching hospital were searched. Multiple linear regression was used to compare independent predictors (patient demographics, anesthetic and intraoperative variables) with the dependent variable (time to extubation). Induction with propofol (P < 0.025) was associated with a shorter time to extubation, while premedication with acepromazine (P = 0.000) was associated with a longer time to extubation. Time to extubation was increased by 0.311 minutes for every kilogram increase in body weight (P = 0.000), 5.924 minutes for every 1 °C loss in body temperature (P = 0.0000), and by 0.096 minutes for every 1 minute increase in anesthetic duration (P = 0.000). Anesthetic variables, which can be manipulated by the anesthetist, include choice of premedication and induction drugs, hypothermia, and duration of anesthesia. Published by Elsevier Ltd.

Control and Analysis for a Self-Excited Induction Generator for Wind Turbine and Electrolyzer Applications

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

Muljadi, Eduard; Na, Woonki; Leighty, Bill

Self-Excited Induction Generation(SEIG) is very rugged, simple, lightweight, and it is easy and inexpensive to implement, very simple to control, and requires a very little maintenance. In this variable-speed operation, the SEIG needs a power electronics interface to convert from the variable frequency output voltage of the generator to a DC output voltage for battery or other DC applications. In our study, a SEIG is connected to the power electronics interface such as diode rectifier and DC/DC converter and then an electrolyzer is connected as a final DC load for fuel cell applications. An equivalent circuit model for an electrolyzermore » is utilized for our application. The control and analysis for the proposed system is carried out by using PSCAD and MATLAB software. This study would be useful for designing and control analysis of power interface circuits for SEIG for a variable speed wind turbine generation with fuel cell applications before the actual implementation.« less

Radiation induction of drug resistance in RIF-1 tumors and tumor cells

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

Hopwood, L.E.; Moulder, J.E.

1989-11-01

The RIF-1 tumor cell line contains a small number of cells (1-20 per 10(6) cells) that are resistant to various single antineoplastic drugs, including 5-fluorouracil (5FU), methotrexate (MTX), and adriamycin (ADR). For 5FU the frequency of drug resistance is lower for tumor-derived cells than for cells from cell culture; for MTX the reverse is true, and for ADR there is no difference. In vitro irradiation at 5 Gy significantly increased the frequency of drug-resistant cells for 5FU, MTX, and ADR. In vivo irradiation at 3 Gy significantly increased the frequency of drug-resistant cells for 5FU and MTX, but not formore » ADR. The absolute risk for in vitro induction of MTX, 5FU, and ADR resistance, and for in vivo induction of 5FU resistance, was 1-3 per 10(6) cells per Gy; but the absolute risk for in vivo induction of MTX resistance was 54 per 10(6) cells per Gy. The frequency of drug-resistant cells among individual untreated tumors was highly variable; among individual irradiated tumors the frequency of drug-resistant cells was significantly less variable. These studies provide supporting data for models of the development of tumor drug resistance, and imply that some of the drug resistance seen when chemotherapy follows radiotherapy may be due to radiation-induced drug resistance.« less

Optimized PID control of depth of hypnosis in anesthesia.

PubMed

Padula, Fabrizio; Ionescu, Clara; Latronico, Nicola; Paltenghi, Massimiliano; Visioli, Antonio; Vivacqua, Giulio

2017-06-01

This paper addresses the use of proportional-integral-derivative controllers for regulating the depth of hypnosis in anesthesia by using propofol administration and the bispectral index as a controlled variable. In fact, introducing an automatic control system might provide significant benefits for the patient in reducing the risk for under- and over-dosing. In this study, the controller parameters are obtained through genetic algorithms by solving a min-max optimization problem. A set of 12 patient models representative of a large population variance is used to test controller robustness. The worst-case performance in the considered population is minimized considering two different scenarios: the induction case and the maintenance case. Our results indicate that including a gain scheduling strategy enables optimal performance for induction and maintenance phases, separately. Using a single tuning to address both tasks may results in a loss of performance up to 102% in the induction phase and up to 31% in the maintenance phase. Further on, it is shown that a suitably designed low-pass filter on the controller output can handle the trade-off between the performance and the noise effect in the control variable. Optimally tuned PID controllers provide a fast induction time with an acceptable overshoot and a satisfactory disturbance rejection performance during maintenance. These features make them a very good tool for comparison when other control algorithms are developed. Copyright © 2017 Elsevier B.V. All rights reserved.

Performance Analysis of Three-Phase Induction Motor with AC Direct and VFD

NASA Astrophysics Data System (ADS)

Kumar, Dinesh

2018-03-01

The electrical machine analysis and performance calculation is a very important aspect of efficient drive system design. The development of power electronics devices and power converters provide smooth speed control of Induction Motors by changing the frequency of input supply. These converters, on one hand are providing a more flexible speed control that also leads to problems of harmonics and their associated ailments like pulsating torque, distorted current and voltage waveforms, increasing losses etc. This paper includes the performance analysis of three phase induction motor with three-phase AC direct and variable frequency drives (VFD). The comparison has been concluded with respect to various parameters. MATLAB-SIMULINKTM is used for the analysis.

Analysis of rainwater dissolved organic carbon compounds using fluorescence spectrophotometry

NASA Astrophysics Data System (ADS)

Muller, Catherine L.; Baker, Andy; Hutchinson, Robert; Fairchild, Ian J.; Kidd, Chris

Global rainwater dissolved organic carbon (DOC) flux was recently estimated as 430 × 10 12 g C yr -1, yet little is known about the wide range of chemical compounds present, their sources, temporal patterns of variation, and the subsequent impact on climate and the environment. Precipitation events were sampled in Birmingham, UK between April 2005 and May 2007. Rainwater DOC compounds were analysed using fluorescence spectrophotometry. Three fluorophores were identified: HUmic-LIke Substances (HULIS), TYrosine-LIke Substances (TYLIS) and TRYptophan-LIke Substances (TRYLIS). Peak fluorescence intensities and locations for each substance were examined, and their variations with various meteorological parameters were investigated. The mean HULIS fluorescence intensity from all events was 209 a.u. (with sample fluorescence ranging from 37 a.u. to 995 a.u); mean fluorescence intensity was 469 a.u. (214-988 a.u) and 265 a.u. (50-876 a.u.) for TYLIS and TRYLIS, respectively. Results indicate that highest HULIS fluorescence intensities are experienced during convective events and events of continental origin, suggesting terrestrial/anthropogenic sources. Under well-mixed conditions, HULIS fluorescence intensity decreases, whereas during low wind speed, stagnation of the atmosphere results in higher fluorescence intensities, attributed to a build up of localised sources, particularly anthropogenic. TYLIS and TRYLIS did not show any significant trends for the meteorological variables. Fluorescence spectrophotometry is a fast, non-invasive technique which is demonstrated to be a powerful means of fingerprinting rainfall DOC compounds in real time for small sample volumes.

VARIABLE TIME DELAY MEANS

DOEpatents

Clemensen, R.E.

1959-11-01

An electrically variable time delay line is described which may be readily controlled simuitaneously with variable impedance matching means coupied thereto such that reflections are prevented. Broadly, the delay line includes a signal winding about a magnetic core whose permeability is electrically variable. Inasmuch as the inductance of the line varies directly with the permeability, the time delay and characteristic impedance of the line both vary as the square root of the permeability. Consequently, impedance matching means may be varied similariy and simultaneously w:th the electrically variable permeability to match the line impedance over the entire range of time delay whereby reflections are prevented.

Monitoring of an antigen manufacturing process.

PubMed

Zavatti, Vanessa; Budman, Hector; Legge, Raymond; Tamer, Melih

2016-06-01

Fluorescence spectroscopy in combination with multivariate statistical methods was employed as a tool for monitoring the manufacturing process of pertactin (PRN), one of the virulence factors of Bordetella pertussis utilized in whopping cough vaccines. Fluorophores such as amino acids and co-enzymes were detected throughout the process. The fluorescence data collected at different stages of the fermentation and purification process were treated employing principal component analysis (PCA). Through PCA, it was feasible to identify sources of variability in PRN production. Then, partial least square (PLS) was employed to correlate the fluorescence spectra obtained from pure PRN samples and the final protein content measured by a Kjeldahl test from these samples. In view that a statistically significant correlation was found between fluorescence and PRN levels, this approach could be further used as a method to predict the final protein content.

The induction of apoptosis by methotrexate in activated lymphocytes as indicated by fluorescence hyperpolarization: a possible model for predicting methotrexate therapy for rheumatoid arthritis patients.

PubMed

Herman, Shoshy; Zurgil, Naomi; Langevitz, Pnina; Ehrenfeld, Michael; Deutsch, Mordechai

2003-04-01

The objectives of this study were to test the in vitro response of healthy non-activated, activated, and rheumatoid arthritis (RA) lymphocytes to methotrexate (MTX), and design an in vitro model for predicting the efficiency of MTX treatment for RA patients. Considering the RA profile of clonal-expanded CD4(+) T cells, phytohemagglutinin-activated mononuclear cells taken from healthy donors were incubated with different concentrations of MTX. The MTX-immunosuppressive effect was tested by fluorescence intensity measurements, including PI assay and annexin V assay. For simple detection, we used the Individual Cell Scanner (IC-S), which enables the measurement of early events in individual cells. Healthy mononuclear cells (MNC), and MNC derived from RA patients, were tested by the IC-S while utilizing fluorescence polarization (FP) measurements of fluorescein diacetate (FDA) as an established marker of activation or suppression. In healthy activated MNC, we found that MTX, through its early incubation period, interferes with the activation signal obtained by PHA and exerts an apoptotic signal, which is noted by increases in the FP. Comparing our model to six long-standing RA patients and five newly-diagnosed patients revealed significant differences in the FP measurements, including fluorescence depolarization as an early established measurement of lymphocyte activation, and hyperpolarization as a measurement of an early immunosuppressive effect. We conclude that MTX, an effective therapy for RA patients, could easily be tested by fluorescence polarization measurements of FDA before (or during) clinical use in order to predict its efficiency on a specific RA patient. Moreover, the FP measurements can be used for the diagnosis, and making timing and dosage decisions.

Pharmacokinetics of Chlorin e6-Cobalt Bis(Dicarbollide) Conjugate in Balb/c Mice with Engrafted Carcinoma

PubMed Central

Volovetsky, Arthur B.; Balalaeva, Irina V.; Dudenkova, Varvara V.; Shilyagina, Natalia Yu.; Feofanov, Аlexey V.; Efremenko, Anastasija V.; Grin, Mikhail A.; Mironov, Andrey F.; Bregadze, Vladimir I.; Maslennikova, Anna V.

2017-01-01

The necessary precondition for efficient boron neutron capture therapy (BNCT) is control over the content of isotope 10B in the tumor and normal tissues. In the case of boron-containing porphyrins, the fluorescent part of molecule can be used for quantitative assessment of the boron content. Study Objective: We performed a study of the biodistribution of the chlorin e6-Cobalt bis(dicarbollide) conjugate in carcinoma-bearing Balb/c mice using ex vivo fluorescence imaging, and developed a mathematical model describing boron accumulation and release based on the obtained experimental data. Materials and Methods: The study was performed on Balb/c tumor-bearing mice (CT-26 tumor model). A solution of the chlorin e6-Cobalt bis(dicarbollide) conjugate (CCDC) was injected into the blood at a dose of 10 mg/kg of the animal’s weight. Analysis of the fluorescence signal intensity was performed at several time points by spectrofluorimetry in blood and by laser scanning microscopy in muscle, liver, and tumor tissues. The boron content in the same samples was determined by mass spectroscopy with inductively coupled plasma. Results: Analysis of a linear approximation between the fluorescence intensity and boron content in the tissues demonstrated a satisfactory value of approximation reliability with a Spearman’s rank correlation coefficient of r = 0.938, p < 0.01. The dynamics of the boron concentration change in various organs, calculated on the basis of the fluorescence intensity, enabled the development of a model describing the accumulation of the studied compound and its distribution in tissues. The obtained results reveal a high level of correspondence between the model and experimental data. PMID:29182594

Pharmacokinetics of Chlorin e₆-Cobalt Bis(Dicarbollide) Conjugate in Balb/c Mice with Engrafted Carcinoma.

PubMed

Volovetsky, Arthur B; Sukhov, Vladimir S; Balalaeva, Irina V; Dudenkova, Varvara V; Shilyagina, Natalia Yu; Feofanov, Аlexey V; Efremenko, Anastasija V; Grin, Mikhail A; Mironov, Andrey F; Sivaev, Igor B; Bregadze, Vladimir I; Maslennikova, Anna V

2017-11-28

The necessary precondition for efficient boron neutron capture therapy (BNCT) is control over the content of isotope 10 B in the tumor and normal tissues. In the case of boron-containing porphyrins, the fluorescent part of molecule can be used for quantitative assessment of the boron content. Study Objective: We performed a study of the biodistribution of the chlorin e ₆-Cobalt bis(dicarbollide) conjugate in carcinoma-bearing Balb/c mice using ex vivo fluorescence imaging, and developed a mathematical model describing boron accumulation and release based on the obtained experimental data. Materials and Methods: The study was performed on Balb/c tumor-bearing mice (CT-26 tumor model). A solution of the chlorin e ₆-Cobalt bis(dicarbollide) conjugate (CCDC) was injected into the blood at a dose of 10 mg/kg of the animal's weight. Analysis of the fluorescence signal intensity was performed at several time points by spectrofluorimetry in blood and by laser scanning microscopy in muscle, liver, and tumor tissues. The boron content in the same samples was determined by mass spectroscopy with inductively coupled plasma. Results: Analysis of a linear approximation between the fluorescence intensity and boron content in the tissues demonstrated a satisfactory value of approximation reliability with a Spearman's rank correlation coefficient of r = 0.938, p < 0.01. The dynamics of the boron concentration change in various organs, calculated on the basis of the fluorescence intensity, enabled the development of a model describing the accumulation of the studied compound and its distribution in tissues. The obtained results reveal a high level of correspondence between the model and experimental data.

ATP hydrolysis is critical for induction of conformational changes in GroEL that expose hydrophobic surfaces.

PubMed

Gorovits, B M; Ybarra, J; Horowitz, P M

1997-03-14

The degree of hydrophobic exposure in the molecular chaperone GroEL during its cycle of ATP hydrolysis was analyzed using 1,1'-bis(4-anilino)naphthalene-5,5'disulfonic acid (bisANS), a hydrophobic probe, whose fluorescence is highly sensitive to the environment. In the presence of 10 mM MgCl2 and 10 mM KCl the addition of ATP, but not ADP or AMP-PNP, resulted in a time-dependent, linear increase in the bisANS fluorescence. The rate of the increase in the bisANS fluorescence depended on the concentrations of both GroEL and the probe. The effect could be substantially inhibited by addition of excess ADP or by converting ATP to ADP using hexokinase, showing that the increase in the bisANS fluorescence was correlated with ATP hydrolysis. The rate of ATP hydrolysis catalyzed by GroEL was uncompetitively inhibited in the presence of bisANS. This uncompetitive inhibition suggests that the probe can interact with the GroEL-ATP complex. The inability of the nonhydrolyzable ATP analog, AMP-PNP, to cause a similar effect is explained by the interaction of bisANS with a transient conformational state of GroEL formed consequent to ATP hydrolysis. It is suggested that this short lived hydrophobic exposure reflects a conformational shift in GroEL that results from electrostatic repulsion between the bound products of ATP hydrolysis, and it plays an important role in the mechanism of the chaperonin cycle.

MIGRATION OF INTRADERMALLY INJECTED QUANTUM DOTS TO SENTINEL ORGANS IN MICE

PubMed Central

Gopee, Neera V.; Roberts, Dean W.; Webb, Peggy; Cozart, Christy R.; Siitonen, Paul H.; Warbritton, Alan R.; Yu, William W.; Colvin, Vicki L.; Walker, Nigel J.; Howard, Paul C.

2012-01-01

Topical exposure to nanoscale materials is likely from a variety of sources including sunscreens and cosmetics. Because the in vivo disposition of nanoscale materials is not well understood, we have evaluated the distribution of quantum dots (QD) following intradermal injection into female SKH-1 hairless mice as a model system for determining tissue localization following intradermal infiltration. The QD [CdSe core, CdS capped, poly(ethylene glycol) (PEG) coated, 37 nm diameter, 621 nm fluorescence emission] were injected intradermally on the right dorsal flank. Within minutes following intradermal injection, the highly UV fluorescent QD could be observed moving from the injection sites apparently through the lymphatic duct system to regional lymph nodes. Residual fluorescent QD remained at the site of injection until necropsy at 24 hours. Quantification of cadmium and selenium levels after 0, 4, 8, 12 or 24 hours in multiple tissues, using inductively coupled plasma mass spectrometry (ICP-MS) showed a time-dependent loss of cadmium from the injection site, and accumulation in the liver, regional draining lymph nodes, kidney, spleen, and hepatic lymph node. Fluorescence microscopy corroborated the ICP-MS results regarding the tissue distribution of QD. The results indicated that (a) intradermally injected nanoscale QD remained as a deposit in skin and penetrated the surrounding viable subcutis, (b) QD were distributed to draining lymph nodes through the subcutaneous lymphatics and to the liver and other organs, and (c) sentinel organs are effective locations for monitoring transdermal penetration of nanoscale materials into animals. PMID:17404394

In Vivo Study of Trichoderma-Pathogen-Plant Interactions, Using Constitutive and Inducible Green Fluorescent Protein Reporter Systems

PubMed Central

Lu, Zexun; Tombolini, Riccardo; Woo, Sheridan; Zeilinger, Susanne; Lorito, Matteo; Jansson, Janet K.

2004-01-01

Plant tissue colonization by Trichoderma atroviride plays a critical role in the reduction of diseases caused by phytopathogenic fungi, but this process has not been thoroughly studied in situ. We monitored in situ interactions between gfp-tagged biocontrol strains of T. atroviride and soilborne plant pathogens that were grown in cocultures and on cucumber seeds by confocal scanning laser microscopy and fluorescence stereomicroscopy. Spores of T. atroviride adhered to Pythium ultimum mycelia in coculture experiments. In mycoparasitic interactions of T. atroviride with P. ultimum or Rhizoctonia solani, the mycoparasitic hyphae grew alongside the pathogen mycelia, and this was followed by coiling and formation of specialized structures similar to hooks, appressoria, and papillae. The morphological changes observed depended on the pathogen tested. Branching of T. atroviride mycelium appeared to be an active response to the presence of the pathogenic host. Mycoparasitism of P. ultimum by T. atroviride occurred on cucumber seed surfaces while the seeds were germinating. The interaction of these fungi on the cucumber seeds was similar to the interaction observed in coculture experiments. Green fluorescent protein expression under the control of host-inducible promoters was also studied. The induction of specific Trichoderma genes was monitored visually in cocultures, on plant surfaces, and in soil in the presence of colloidal chitin or Rhizoctonia by confocal microscopy and fluorescence stereomicroscopy. These tools allowed initiation of the mycoparasitic gene expression cascade to be monitored in vivo. PMID:15128569

The influence of indoxyl sulfate and ammonium on the autofluorescence of human urine.

PubMed

Perinchery, Sandeep Menon; Kuzhiumparambil, Unnikrishnan; Vemulpad, Subramanyam; Goldys, Ewa M

2010-01-15

Despite biological variability the spectral characteristics of undiluted human urine show relatively low autofluorescence at short UV (250-300nm) excitation. However with dilution the fluorescence intensity remarkably increases. This paper examines the mechanisms behind this effect, by using excitation-emission matrices. Corrections for the inner filter effect were made for improved understanding of the spectral patterns. We focused on three major fluorophores (tryptophan, indoxyl sulfate and 5-hydroxyindole-3-acetate) that are excited at these wavelengths, and whose content in urine is strongly linked with various health conditions. Their fluorescence was studied both individually and in combinations. We also examined the effect of ammonium on the fluorescence of these major fluorophores individually and in combinations. Through these studies we have identified the leading effects that reduce the UV fluorescence, namely higher concentration of indoxyl sulfate producing the inner filter effect and concentration quenching and quenching of fluorophores by ammonium. This result will assist in broader utilisation of UV fluorescence in medical diagnostics.

Simple device for the direct visualization of oral-cavity tissue fluorescence

NASA Astrophysics Data System (ADS)

Lane, Pierre M.; Gilhuly, Terence; Whitehead, Peter D.; Zeng, Haishan; Poh, Catherine; Ng, Samson; Williams, Michelle; Zhang, Lewei; Rosin, Miriam; MacAulay, Calum E.

2006-03-01

Early identification of high-risk disease could greatly reduce both mortality and morbidity due to oral cancer. We describe a simple handheld device that facilitates the direct visualization of oral-cavity fluorescence for the detection of high-risk precancerous and early cancerous lesions. Blue excitation light (400 to 460 nm) is employed to excite green-red fluorescence from fluorophores in the oral tissues. Tissue fluorescence is viewed directly along an optical axis collinear with the axis of excitation to reduce inter- and intraoperator variability. This robust, field-of-view device enables the direct visualization of fluorescence in the context of surrounding normal tissue. Results from a pilot study of 44 patients are presented. Using histology as the gold standard, the device achieves a sensitivity of 98% and specificity of 100% when discriminating normal mucosa from severe dysplasia/carcinoma in situ (CIS) or invasive carcinoma. We envisage this device as a suitable adjunct for oral cancer screening, biopsy guidance, and margin delineation.

Comparative assessment of fluorescent proteins for in vivo imaging in an animal model system.

PubMed

Heppert, Jennifer K; Dickinson, Daniel J; Pani, Ariel M; Higgins, Christopher D; Steward, Annette; Ahringer, Julie; Kuhn, Jeffrey R; Goldstein, Bob

2016-11-07

Fluorescent protein tags are fundamental tools used to visualize gene products and analyze their dynamics in vivo. Recent advances in genome editing have expedited the precise insertion of fluorescent protein tags into the genomes of diverse organisms. These advances expand the potential of in vivo imaging experiments and facilitate experimentation with new, bright, photostable fluorescent proteins. Most quantitative comparisons of the brightness and photostability of different fluorescent proteins have been made in vitro, removed from biological variables that govern their performance in cells or organisms. To address the gap, we quantitatively assessed fluorescent protein properties in vivo in an animal model system. We generated transgenic Caenorhabditis elegans strains expressing green, yellow, or red fluorescent proteins in embryos and imaged embryos expressing different fluorescent proteins under the same conditions for direct comparison. We found that mNeonGreen was not as bright in vivo as predicted based on in vitro data but is a better tag than GFP for specific kinds of experiments, and we report on optimal red fluorescent proteins. These results identify ideal fluorescent proteins for imaging in vivo in C. elegans embryos and suggest good candidate fluorescent proteins to test in other animal model systems for in vivo imaging experiments. © 2016 Heppert et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

Genotype-Specific Minimal Residual Disease Interpretation Improves Stratification in Pediatric Acute Lymphoblastic Leukemia

PubMed Central

O’Connor, David; Enshaei, Amir; Bartram, Jack; Hancock, Jeremy; Harrison, Christine J.; Hough, Rachael; Samarasinghe, Sujith; Schwab, Claire; Vora, Ajay; Wade, Rachel; Moppett, John; Moorman, Anthony V.; Goulden, Nick

2018-01-01

Purpose Minimal residual disease (MRD) and genetic abnormalities are important risk factors for outcome in acute lymphoblastic leukemia. Current risk algorithms dichotomize MRD data and do not assimilate genetics when assigning MRD risk, which reduces predictive accuracy. The aim of our study was to exploit the full power of MRD by examining it as a continuous variable and to integrate it with genetics. Patients and Methods We used a population-based cohort of 3,113 patients who were treated in UKALL2003, with a median follow-up of 7 years. MRD was evaluated by polymerase chain reaction analysis of Ig/TCR gene rearrangements, and patients were assigned to a genetic subtype on the basis of immunophenotype, cytogenetics, and fluorescence in situ hybridization. To examine response kinetics at the end of induction, we log-transformed the absolute MRD value and examined its distribution across subgroups. Results MRD was log normally distributed at the end of induction. MRD distributions of patients with distinct genetic subtypes were different (P < .001). Patients with good-risk cytogenetics demonstrated the fastest disease clearance, whereas patients with high-risk genetics and T-cell acute lymphoblastic leukemia responded more slowly. The risk of relapse was correlated with MRD kinetics, and each log reduction in disease level reduced the risk by 20% (hazard ratio, 0.80; 95% CI, 0.77 to 0.83; P < .001). Although the risk of relapse was directly proportional to the MRD level within each genetic risk group, absolute relapse rate that was associated with a specific MRD value or category varied significantly by genetic subtype. Integration of genetic subtype–specific MRD values allowed more refined risk group stratification. Conclusion A single threshold for assigning patients to an MRD risk group does not reflect the response kinetics of the different genetic subtypes. Future risk algorithms should integrate genetics with MRD to accurately identify patients with the lowest and highest risk of relapse. PMID:29131699

Predictors of self-reported negative mood following a depressive mood induction procedure across previously depressed, currently anxious, and control individuals.

PubMed

Scherrer, Martin C; Dobson, Keith S; Quigley, Leanne

2014-09-01

This study identified and examined a set of potential predictors of self-reported negative mood following a depressive mood induction procedure (MIP) in a sample of previously depressed, clinically anxious, and control participants. The examined predictor variables were selected on the basis of previous research and theories of depression, and included symptoms of depression and anxiety, negative and positive affect, negative and positive automatic thoughts, dysfunctional beliefs, rumination, self-concept, and occurrence and perceived unpleasantness of recent negative events. The sample consisted of 33 previously depressed, 22 currently anxious, and 26 non-clinical control participants, recruited from community sources. Participant group status was confirmed through structured diagnostic interviews. Participants completed the Velten negative self-statement MIP as well as self-report questionnaires of affective, cognitive, and psychosocial variables selected as potential predictors of mood change. Symptoms of anxiety were associated with increased self-reported negative mood shift following the MIP in previously depressed participants, but not clinically anxious or control participants. Increased occurrence of recent negative events was a marginally significant predictor of negative mood shift for the previously depressed participants only. None of the other examined variables was significant predictors of MIP response for any of the participant groups. These results identify factors that may increase susceptibility to negative mood states in previously depressed individuals, with implications for theory and prevention of relapse to depression. The findings also identify a number of affective, cognitive, and psychosocial variables that do not appear to influence mood change following a depressive MIP in previously depressed, currently anxious, and control individuals. Limitations of the study and directions for future research are discussed. Current anxiety symptomatology was a significant predictor and occurrence of recent negative events was a marginally significant predictor of greater negative mood shift following the depressive mood induction for previously depressed individuals. None of the examined variables predicted change in mood following the depressive mood induction for currently anxious or control individuals. These results suggest that anxiety symptoms and experience with negative events may increase risk for experiencing depressive mood states among individuals with a vulnerability to depression. The generalizability of the present results to individuals with comorbid depression and anxiety is limited. Future research employing appropriate statistical approaches for confirmatory research is needed to test and confirm the present results. © 2014 The British Psychological Society.

Investigating the procedural variables that determine whether rats will display negative anticipatory contrast or positive induction.

PubMed

Weatherly, Jeffrey N; Nurnberger, Jeri T; Hanson, Brent C

2005-08-31

Previous studies have demonstrated that consumption of a low-valued food substance may decrease if access to a high-valued substance will soon be available (negative anticipatory contrast). Research has also demonstrated that responding for a low-valued reinforcer may increase if responding for a high-valued reinforcer will soon be possible (positive induction). The present experiment employed rats to respond in a procedure similar to that typically used to produce negative anticipatory contrast. The goal was to determine what factors contribute to when a contrast or an induction effect will occur. Based on previous research, the influence of auditory cues, temporal delays, food deprivation, and location of substance delivery were investigated. Auditory cues and temporal delays did little to influence whether subjects increased or decreased their consumption of 1% sucrose when access to 32% sucrose was upcoming. The appearance of contrast or induction was related to level of deprivation, with deprivation promoting induction. Which effect occurred also depended on whether subjects consumed the two substances from one spout in one location (induction) or from two different spouts in two different locations (contrast). The present results help identify the procedural link(s) between these two effects. They also provide insight to why positive induction may occur (i.e., higher-order place conditioning).

Mapping Copper and Lead Concentrations at Abandoned Mine Areas Using Element Analysis Data from ICP–AES and Portable XRF Instruments: A Comparative Study

PubMed Central

Lee, Hyeongyu; Choi, Yosoon; Suh, Jangwon; Lee, Seung-Ho

2016-01-01

Understanding spatial variation of potentially toxic trace elements (PTEs) in soil is necessary to identify the proper measures for preventing soil contamination at both operating and abandoned mining areas. Many studies have been conducted worldwide to explore the spatial variation of PTEs and to create soil contamination maps using geostatistical methods. However, they generally depend only on inductively coupled plasma atomic emission spectrometry (ICP–AES) analysis data, therefore such studies are limited by insufficient input data owing to the disadvantages of ICP–AES analysis such as its costly operation and lengthy period required for analysis. To overcome this limitation, this study used both ICP–AES and portable X-ray fluorescence (PXRF) analysis data, with relatively low accuracy, for mapping copper and lead concentrations at a section of the Busan abandoned mine in Korea and compared the prediction performances of four different approaches: the application of ordinary kriging to ICP–AES analysis data, PXRF analysis data, both ICP–AES and transformed PXRF analysis data by considering the correlation between the ICP–AES and PXRF analysis data, and co-kriging to both the ICP–AES (primary variable) and PXRF analysis data (secondary variable). Their results were compared using an independent validation data set. The results obtained in this case study showed that the application of ordinary kriging to both ICP–AES and transformed PXRF analysis data is the most accurate approach when considers the spatial distribution of copper and lead contaminants in the soil and the estimation errors at 11 sampling points for validation. Therefore, when generating soil contamination maps for an abandoned mine, it is beneficial to use the proposed approach that incorporates the advantageous aspects of both ICP–AES and PXRF analysis data. PMID:27043594

Mapping Copper and Lead Concentrations at Abandoned Mine Areas Using Element Analysis Data from ICP-AES and Portable XRF Instruments: A Comparative Study.

PubMed

Lee, Hyeongyu; Choi, Yosoon; Suh, Jangwon; Lee, Seung-Ho

2016-03-30

Understanding spatial variation of potentially toxic trace elements (PTEs) in soil is necessary to identify the proper measures for preventing soil contamination at both operating and abandoned mining areas. Many studies have been conducted worldwide to explore the spatial variation of PTEs and to create soil contamination maps using geostatistical methods. However, they generally depend only on inductively coupled plasma atomic emission spectrometry (ICP-AES) analysis data, therefore such studies are limited by insufficient input data owing to the disadvantages of ICP-AES analysis such as its costly operation and lengthy period required for analysis. To overcome this limitation, this study used both ICP-AES and portable X-ray fluorescence (PXRF) analysis data, with relatively low accuracy, for mapping copper and lead concentrations at a section of the Busan abandoned mine in Korea and compared the prediction performances of four different approaches: the application of ordinary kriging to ICP-AES analysis data, PXRF analysis data, both ICP-AES and transformed PXRF analysis data by considering the correlation between the ICP-AES and PXRF analysis data, and co-kriging to both the ICP-AES (primary variable) and PXRF analysis data (secondary variable). Their results were compared using an independent validation data set. The results obtained in this case study showed that the application of ordinary kriging to both ICP-AES and transformed PXRF analysis data is the most accurate approach when considers the spatial distribution of copper and lead contaminants in the soil and the estimation errors at 11 sampling points for validation. Therefore, when generating soil contamination maps for an abandoned mine, it is beneficial to use the proposed approach that incorporates the advantageous aspects of both ICP-AES and PXRF analysis data.

Development of fiber optic spectroscopy for in-vitro and in-planta detection of fluorescent proteins

NASA Astrophysics Data System (ADS)

Liew, Oi Wah; Chen, Jun-Wei; Asundi, Anand K.

2001-10-01

The objective of this project is to apply photonics technology to bio-safety management of genetically modified (GM) plants. The conventional method for screening GM plants is through selection using antibiotic resistance markers. There is public concern with such approaches and these are associated with food safety issues, escape of antibiotic resistance genes to pathogenic microorganisms and interference with antibiotic therapy. Thus, the strategy taken in this project is to replace antibiotic resistance markers with fluorescent protein markers that allow for rapid and non-invasive optical screening of genetically modified plants. In this paper, fibre optic spectroscopy was developed to detect and quantify recombinant green (EGFP) and red (DsRED) fluorescent proteins in vitro and in planta. In vitro detection was first carried out to optimize the sensitivity of the optical system. The bacterial expression vectors carrying the coding regions of EGFP and DsRED were introduced into Escherichia coli host cells and fluorescent proteins were produced following induction with IPTG. Soluble EGFP and DsRED proteins were isolated from lysed bacterial cells and serially diluted for quantitative analysis by fibre optic spectroscopy using different light sources, namely, blue LED (475 nm), tungsten halogen (350 - 1000 nm) and double frequency Nd:YAG green laser (532 nm). Fluorescence near the expected emission wavelengths could be detected up to 320X dilution for EGFP and DsRED with blue LED and 532 nm green laser, respectively, as the excitation source. Tungsten halogen was found to be unsuitable for excitation of both EGFP and DsRED. EGFP was successfully purified by size separation under non-denaturing electrophoretic conditions and quantified. The minimum concentration of EGFP detectable with blue LED excitation was 5 mg/ml. To determine the capability of spectroscopy detection in planta, transgenic potato hairy roots and whole modified plant lines expressing the fluorescent markers were regenerated. T

Picturing pathogen infection in plants.

PubMed

Barón, Matilde; Pineda, Mónica; Pérez-Bueno, María Luisa

2016-09-01

Several imaging techniques have provided valuable tools to evaluate the impact of biotic stress on host plants. The use of these techniques enables the study of plant-pathogen interactions by analysing the spatial and temporal heterogeneity of foliar metabolism during pathogenesis. In this work we review the use of imaging techniques based on chlorophyll fluorescence, multicolour fluorescence and thermography for the study of virus, bacteria and fungi-infected plants. These studies have revealed the impact of pathogen challenge on photosynthetic performance, secondary metabolism, as well as leaf transpiration as a promising tool for field and greenhouse management of diseases. Images of standard chlorophyll fluorescence (Chl-F) parameters obtained during Chl-F induction kinetics related to photochemical processes and those involved in energy dissipation, could be good stress indicators to monitor pathogenesis. Changes on UV-induced blue (F440) and green fluorescence (F520) measured by multicolour fluorescence imaging in pathogen-challenged plants seem to be related with the up-regulation of the plant secondary metabolism and with an increase in phenolic compounds involved in plant defence, such as scopoletin, chlorogenic or ferulic acids. Thermal imaging visualizes the leaf transpiration map during pathogenesis and emphasizes the key role of stomata on innate plant immunity. Using several imaging techniques in parallel could allow obtaining disease signatures for a specific pathogen. These techniques have also turned out to be very useful for presymptomatic pathogen detection, and powerful non-destructive tools for precision agriculture. Their applicability at lab-scale, in the field by remote sensing, and in high-throughput plant phenotyping, makes them particularly useful. Thermal sensors are widely used in crop fields to detect early changes in leaf transpiration induced by both air-borne and soil-borne pathogens. The limitations of measuring photosynthesis by Chl-F at the canopy level are being solved, while the use of multispectral fluorescence imaging is very challenging due to the type of light excitation that is used.

Classification of different kinds of pesticide residues on lettuce based on fluorescence spectra and WT-BCC-SVM algorithm

NASA Astrophysics Data System (ADS)

Zhou, Xin; Jun, Sun; Zhang, Bing; Jun, Wu

2017-07-01

In order to improve the reliability of the spectrum feature extracted by wavelet transform, a method combining wavelet transform (WT) with bacterial colony chemotaxis algorithm and support vector machine (BCC-SVM) algorithm (WT-BCC-SVM) was proposed in this paper. Besides, we aimed to identify different kinds of pesticide residues on lettuce leaves in a novel and rapid non-destructive way by using fluorescence spectra technology. The fluorescence spectral data of 150 lettuce leaf samples of five different kinds of pesticide residues on the surface of lettuce were obtained using Cary Eclipse fluorescence spectrometer. Standard normalized variable detrending (SNV detrending), Savitzky-Golay coupled with Standard normalized variable detrending (SG-SNV detrending) were used to preprocess the raw spectra, respectively. Bacterial colony chemotaxis combined with support vector machine (BCC-SVM) and support vector machine (SVM) classification models were established based on full spectra (FS) and wavelet transform characteristics (WTC), respectively. Moreover, WTC were selected by WT. The results showed that the accuracy of training set, calibration set and the prediction set of the best optimal classification model (SG-SNV detrending-WT-BCC-SVM) were 100%, 98% and 93.33%, respectively. In addition, the results indicated that it was feasible to use WT-BCC-SVM to establish diagnostic model of different kinds of pesticide residues on lettuce leaves.

Approximate analytical solution for induction heating of solid cylinders

DOE PAGES

Jankowski, Todd Andrew; Pawley, Norma Helen; Gonzales, Lindsey Michal; ...

2015-10-20

An approximate solution to the mathematical model for induction heating of a solid cylinder in a cylindrical induction coil is presented here. The coupled multiphysics model includes equations describing the electromagnetic field in the heated object, a heat transfer simulation to determine temperature of the heated object, and an AC circuit simulation of the induction heating power supply. A multiple-scale perturbation method is used to solve the multiphysics model. The approximate analytical solution yields simple closed-form expressions for the electromagnetic field and heat generation rate in the solid cylinder, for the equivalent impedance of the associated tank circuit, and formore » the frequency response of a variable frequency power supply driving the tank circuit. The solution developed here is validated by comparing predicted power supply frequency to both experimental measurements and calculated values from finite element analysis for heating of graphite cylinders in an induction furnace. The simple expressions from the analytical solution clearly show the functional dependence of the power supply frequency on the material properties of the load and the geometrical characteristics of the furnace installation. In conclusion, the expressions developed here provide physical insight into observations made during load signature analysis of induction heating.« less

Defining the Ideal Time Interval Between Planned Induction Therapy and Surgery for Stage IIIA Non-Small Cell Lung Cancer.

PubMed

Samson, Pamela; Crabtree, Traves D; Robinson, Cliff G; Morgensztern, Daniel; Broderick, Stephen; Krupnick, A Sasha; Kreisel, Daniel; Patterson, G Alexander; Meyers, Bryan; Puri, Varun

2017-04-01

Induction therapy leads to significant improvement in survival for selected patients with stage IIIA non-small cell lung cancer. The ideal time interval between induction therapy and surgery remains unknown. Clinical stage IIIA non-small cell lung cancer patients receiving induction therapy and surgery were identified in the National Cancer Database. Delayed surgery was defined as greater than or equal to 3 months after starting induction therapy. A logistic regression model identified variables associated with delayed surgery. Cox proportional hazards modeling and Kaplan-Meier analysis were performed to evaluate variables independently associated with overall survival. From 2006 to 2010, 1,529 of 2,380 (64.2%) received delayed surgery. Delayed surgery patients were older (61.2 ± 10.0 years versus 60.3 ± 9.2; p = 0.03), more likely to be non-white (12.4% versus 9.7%; p = 0.046), and less likely to have private insurance (50% versus 58.2%; p = 0.002). Delayed surgery patients were also more likely to have a sublobar resection (6.3% versus 2.9%). On multivariate analysis, age greater than 68 years (odds ratio [OR], 1.37; 95% confidence interval [CI], 1.1 to 1.7) was associated with delayed surgery, whereas white race (OR, 0.75; 95% CI, 0.57 to 0.99) and private insurance status (OR, 0.82; 95% CI, 0.68 to 0.99) were associated with early surgery. Delayed surgery was associated with higher risk of long-term mortality (hazard ratio, 1.25; 95% CI, 1.07 to 1.47). Delayed surgery after induction therapy for stage IIIA lung cancer is associated with shorter survival, and is influenced by both social and physiologic factors. Prospective work is needed to further characterize the relationship between patient comorbidities and functional status with receipt of timely surgery. Copyright © 2017 The Society of Thoracic Surgeons. Published by Elsevier Inc. All rights reserved.

Adaptive control in series load PWM induction heating inverters

NASA Astrophysics Data System (ADS)

Szelitzky, Tibor; Henrietta Dulf, Eva

2013-12-01

Permanent variations of the electric properties of the load in induction heating equipment make difficult to control the plant. To overcome these disadvantages, the authors propose a new approach based on adaptive control methods. For real plants it is enough to present desired performances or start-up variables for the controller, from which the algorithms tune the controllers by itself. To present the advantages of the proposed controllers, comparisons are made to a PI controller tuned through Ziegler-Nichols method.

Use of an anaesthesia workstation barrier device to decrease contamination in a simulated operating room.

PubMed

Hunter, S; Katz, D; Goldberg, A; Lin, H-M; Pasricha, R; Benesh, G; Le Grand, B; DeMaria, S

2017-06-01

Strategies to achieve reductions in perioperative infections have focused on hand hygiene among anaesthestists but have been of limited efficacy. We performed a study in a simulated operating room to determine whether a barrier covering the anaesthesia workstation during induction and intubation might reduce the risk of contamination of the area and possibly, by extension, the patient. Forty-two attending and resident anaesthetists unaware of the study design were enrolled in individual simulation sessions in which they were asked to induce and intubate a human simulator that had been prepared with fluorescent marker in its oropharynx as a marker of potentially pathogenic bacteria. Twenty-one participants were assigned to a control group, whereas the other 21 performed the simulation with a barrier device covering the anaesthesia workstation. After the simulation, an investigator examined 14 target sites with an ultraviolet light to assess spread of the fluorescent marker of contamination to those sites. The difference in rates of contamination between the control group and the barrier group was highly significant, with 44.8% (2.5%) of sites contaminated in the control group vs 19.4% (2.6%) of sites in the barrier group ( P <0.001). Several key clinical sites showed significant differences in addition to this overall decrement. The results of this study suggest that application of a barrier device to the anaesthesia workstation during induction and intubation might reduce contamination of the intraoperative environment. © The Author 2017. Published by Oxford University Press on behalf of the British Journal of Anaesthesia. All rights reserved. For Permissions, please email: journals.permissions@oup.com

Assessment of the contamination problems resulting from the use of stainless steel needles in liver biopsies by total reflection X-ray fluorescence and inductively coupled plasma mass spectrometry

NASA Astrophysics Data System (ADS)

Varga, Imre

2006-11-01

Percutaneous human liver biopsies taken from living patients could not be repeated; therefore considerable contamination was indirectly disproved. In the present study, the possible contamination of biopsy samples during sample collection was determined using a porcine liver model. Portions of porcine liver were cut by a quartz blade and treated the same as the steel needle biopsy samples. Concentrations determined in samples taken by a quartz device represented the non-contaminated values and were used to determine reproducibility of measurement and intra-individual variations. Additionally, multiple samples taken from a human liver of a patient suffering steatosis during autopsy were used to determine intra-individual variation of element concentrations. Concentration data of non-contaminated samples were compared to data of steel needle biopsy samples. To investigate the possible release of elements from the steel needles the samples were allowed to contact with the needle for different time in a refrigerator at 4 °C. Total reflection X-ray fluorescence spectrometry (TXRF) and inductively coupled plasma-mass spectrometry (ICP-MS) were applied for simultaneous determination of Cr, Mn, Fe, Co, Ni, Cu, Zn, Rb, Mo and Pb because of the very low sample demand of the two selected techniques. Although the steel needles in the present study could not be substituted by polypropylene or Teflon utensils, it was demonstrated that the application of needle biopsy sampling in the reported analysis does not involve measurable contamination if contact time is kept to several minutes as usual in the clinical practice.

Rapid screening of heavy metals and trace elements in environmental samples using portable X-ray fluorescence spectrometer, A comparative study

PubMed Central

McComb, Jacqueline Q.; Rogers, Christian; Han, Fengxiang X.; Tchounwou, Paul B.

2014-01-01

With industrialization, great amounts of trace elements and heavy metals have been excavated and released on the surface of the earth and dissipated into the environments. Rapid screening technology for detecting major and trace elements as well as heavy metals in variety of environmental samples is most desired. The objectives of this study were to determine the detection limits, accuracy, repeatability and efficiency of a X-ray fluorescence spectrometer (Niton XRF analyzer) in comparison with the traditional analytical methods, inductively coupled plasma optical emission spectrometer (ICP-OES) and inductively coupled plasma optical emission spectrometer (ICP-MS) in screening of major and trace elements of environmental samples including estuary soils and sediments, contaminated soils, and biological samples. XRF is a fast and non-destructive method in measuring the total concentration of multi--elements simultaneously. Contrary to ICP-OES and ICP-MS, XRF analyzer is characterized by the limited preparation required for solid samples, non-destructive analysis, increased total speed and high throughout, the decreased production of hazardous waste and the low running costs as well as multi-elemental determination and portability in the fields. The current comparative study demonstrates that XRF is a good rapid non-destructive method for contaminated soils, sediments and biological samples containing higher concentrations of major and trace elements. Unfortunately, XRF does not have sensitive detection limits of most major and trace elements as ICP-OES or ICP-MS but it may serve as a rapid screening tool for locating hot spots of uncontaminated field soils and sediments. PMID:25861136

Comparison of a portable micro-X-ray fluorescence spectrometry with inductively coupled plasma atomic emission spectrometry for the ancient ceramics analysis

NASA Astrophysics Data System (ADS)

Papadopoulou, D. N.; Zachariadis, G. A.; Anthemidis, A. N.; Tsirliganis, N. C.; Stratis, J. A.

2004-12-01

Two multielement instrumental methods of analysis, micro X-ray fluorescence spectrometry (micro-XRF) and inductively coupled plasma atomic emission spectrometry (ICP-AES) were applied for the analysis of 7th and 5th century B.C. ancient ceramic sherds in order to evaluate the above two methods and to assess the potential to use the current compact and portable micro-XRF instrument for the in situ analysis of ancient ceramics. The distinguishing factor of interest is that micro-XRF spectrometry offers the possibility of a nondestructive analysis, an aspect of primary importance in the compositional analysis of cultural objects. Micro-XRF measurements were performed firstly directly on the ceramic sherds with no special pretreatment apart from surface cleaning (micro-XRF on sherds) and secondly on pressed pellet disks which were prepared for each ceramic sherd (micro-XRF on pellet). For the ICP-AES determination of elements, test solutions were prepared by the application of a microwave-assisted decomposition procedure in closed high-pressure PFA vessels. Also, the standard reference material SARM 69 was used for the efficiency calibration of the micro-XRF instrument and was analysed by both methods. In order to verify the calibration, the standard reference materials NCS DC 73332 and SRM620 as well as the reference materials AWI-1 and PRI-1 were analysed by micro-XRF. Elemental concentrations determined by the three analytical procedures (ICP-AES, micro-XRF on sherds and micro-XRF on pellets) were statistically treated by correlation analysis and Student's t-test (at the 95% confidence level).

Event-Based control of depth of hypnosis in anesthesia.

PubMed

Merigo, Luca; Beschi, Manuel; Padula, Fabrizio; Latronico, Nicola; Paltenghi, Massimiliano; Visioli, Antonio

2017-08-01

In this paper, we propose the use of an event-based control strategy for the closed-loop control of the depth of hypnosis in anesthesia by using propofol administration and the bispectral index as a controlled variable. A new event generator with high noise-filtering properties is employed in addition to a PIDPlus controller. The tuning of the parameters is performed off-line by using genetic algorithms by considering a given data set of patients. The effectiveness and robustness of the method is verified in simulation by implementing a Monte Carlo method to address the intra-patient and inter-patient variability. A comparison with a standard PID control structure shows that the event-based control system achieves a reduction of the total variation of the manipulated variable of 93% in the induction phase and of 95% in the maintenance phase. The use of event based automatic control in anesthesia yields a fast induction phase with bounded overshoot and an acceptable disturbance rejection. A comparison with a standard PID control structure shows that the technique effectively mimics the behavior of the anesthesiologist by providing a significant decrement of the total variation of the manipulated variable. Copyright © 2017 Elsevier B.V. All rights reserved.

Controlling false-negative errors in microarray differential expression analysis: a PRIM approach.

PubMed

Cole, Steve W; Galic, Zoran; Zack, Jerome A

2003-09-22

Theoretical considerations suggest that current microarray screening algorithms may fail to detect many true differences in gene expression (Type II analytic errors). We assessed 'false negative' error rates in differential expression analyses by conventional linear statistical models (e.g. t-test), microarray-adapted variants (e.g. SAM, Cyber-T), and a novel strategy based on hold-out cross-validation. The latter approach employs the machine-learning algorithm Patient Rule Induction Method (PRIM) to infer minimum thresholds for reliable change in gene expression from Boolean conjunctions of fold-induction and raw fluorescence measurements. Monte Carlo analyses based on four empirical data sets show that conventional statistical models and their microarray-adapted variants overlook more than 50% of genes showing significant up-regulation. Conjoint PRIM prediction rules recover approximately twice as many differentially expressed transcripts while maintaining strong control over false-positive (Type I) errors. As a result, experimental replication rates increase and total analytic error rates decline. RT-PCR studies confirm that gene inductions detected by PRIM but overlooked by other methods represent true changes in mRNA levels. PRIM-based conjoint inference rules thus represent an improved strategy for high-sensitivity screening of DNA microarrays. Freestanding JAVA application at http://microarray.crump.ucla.edu/focus

pH-Responsive Micelle-Based Cytoplasmic Delivery System for Induction of Cellular Immunity.

PubMed

Yuba, Eiji; Sakaguchi, Naoki; Kanda, Yuhei; Miyazaki, Maiko; Koiwai, Kazunori

2017-11-04

(1) Background: Cytoplasmic delivery of antigens is crucial for the induction of cellular immunity, which is an important immune response for the treatment of cancer and infectious diseases. To date, fusogenic protein-incorporated liposomes and pH-responsive polymer-modified liposomes have been used to achieve cytoplasmic delivery of antigen via membrane rupture or fusion with endosomes. However, a more versatile cytoplasmic delivery system is desired for practical use. For this study, we developed pH-responsive micelles composed of dilauroyl phosphatidylcholine (DLPC) and deoxycholic acid and investigated their cytoplasmic delivery performance and immunity-inducing capability. (2) Methods: Interaction of micelles with fluorescence dye-loaded liposomes, intracellular distribution of micelles, and antigenic proteins were observed. Finally, antigen-specific cellular immune response was evaluated in vivo using ELIspot assay. (3) Results: Micelles induced leakage of contents from liposomes via lipid mixing at low pH. Micelles were taken up by dendritic cells mainly via macropinocytosis and delivered ovalbumin (OVA) into the cytosol. After intradermal injection of micelles and OVA, OVA-specific cellular immunity was induced in the spleen. (4) Conclusions: pH-responsive micelles composed of DLPC and deoxycholic acid are promising as enhancers of cytosol delivery of antigens and the induction capability of cellular immunity for the treatment of cancer immunotherapy and infectious diseases.

Laminins affect T cell trafficking and allograft fate

PubMed Central

Warren, Kristi J.; Iwami, Daiki; Harris, Donald G.; Bromberg, Jonathan S.; Burrell, Bryna E.

2014-01-01

Lymph nodes (LNs) are integral sites for the generation of immune tolerance, migration of CD4+ T cells, and induction of Tregs. Despite the importance of LNs in regulation of inflammatory responses, the LN-specific factors that regulate T cell migration and the precise LN structural domains in which differentiation occurs remain undefined. Using intravital and fluorescent microscopy, we found that alloreactive T cells traffic distinctly into the tolerant LN and colocalize in exclusive regions with alloantigen-presenting cells, a process required for Treg induction. Extracellular matrix proteins, including those of the laminin family, formed regions within the LN that were permissive for colocalization of alloantigen-presenting cells, alloreactive T cells, and Tregs. We identified unique expression patterns of laminin proteins in high endothelial venule basement membranes and the cortical ridge that correlated with alloantigen-specific immunity or immune tolerance. The ratio of laminin α4 to laminin α5 was greater in domains within tolerant LNs, compared with immune LNs, and blocking laminin α4 function or inducing laminin α5 overexpression disrupted T cell and DC localization and transmigration through tolerant LNs. Furthermore, reducing α4 laminin circumvented tolerance induction and induced cardiac allograft inflammation and rejection in murine models. This work identifies laminins as potential targets for immune modulation. PMID:24691446

Primula auriculata Extracts Exert Cytotoxic and Apoptotic Effects against HT-29 Human Colon Adenocarcinoma Cells

PubMed Central

Behzad, Sahar; Ebrahim, Karim; Mosaddegh, Mahmoud; Haeri, Ali

2016-01-01

Primula auriculata (Tootia) is one of the most important local medicinal plants in Hamedan district, Iran. To investigate cytotoxicity and apoptosis induction of crude methanolic extract and different fraction of it, we compared several methods on HT-29 human colon Adenocarcinoma cells. Cancer cell proliferation was measured by 3-(4, 5‑dimethylthiazolyl)2, 5‑diphenyl‑tetrazolium bromide (MTT) assay and apoptosis induction was analyzed by fluorescence microscopy (acridin orange/ethidium bromide, annexin V/propidium iodide staining, TUNEL assay and Caspase-3 activity assay). Crude methanolic extract (CM) inhibited the growth of malignant cells in a dose-dependent manner. Among solvent fractions, the dichloromethane fraction (CF) was found to be the most toxic compared to other fractions. With double staining methods, high percentage of 40 µg/mL of (CM) and (CF) treated cells exhibited typical characteristics of apoptotic cells. Apoptosis induction was also revealed by apoptotic fragmentation of nuclear DNA and activation of caspas-3 in treated cells. These findings indicate that crude methanolic extract and dichloromethan fraction of P.auriculata induced apoptosis and inhibited proliferation in colon cancer cells and could be used as a source for new lead structures in drug design to combat colon cancer. PMID:27610172

Apoptotic induction of skin cancer cell death by plant extracts.

PubMed

Thuncharoen, Walairat; Chulasiri, Malin; Nilwarangkoon, Sirinun; Nakamura, Yukio; Watanapokasin, Ramida

2013-01-01

The aim of the present study was to investigate the effects of plant extracts on cancer apoptotic induction. Human epidermoid carcinoma A431 cell line, obtained from the American Type Culture Collection (ATCC, Manassas, VA), was maintained in Dulbecco's Modified Eagle Medium (DMEM) supplemented with 10% fetal bovine serum (FBS) at 37 degrees C, 5% carbon dioxide (CO2). Plant extract solutions were obtained from S & J international enterprises public company limited. These plant extracts include 50% hydroglycol extracts from Etlingera elatior (Jack) R.M.Smith (torch ginger; EE), Rosa damascene (damask rose; DR) and Rafflesia kerrii Meijer (bua phut; RM). The cell viability, time and dose dependency were determined by MTT (3-(4, 5-Dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide) assay. A431 cells were treated with the plant extracts and stained with Hoechst 33342 fluorescent staining dye. Cell viability was demonstrated by the inhibitory concentration 50% (IC50). The anti-proliferative effects were shown to be dependent on time and dose. Typical characteristics of apoptosis which are cell morphological changes and chromatin condensation were clearly observed. The plant extracts was shown to be effective for anti-proliferation and induction of apoptosis cell death in skin cancer cells. Therefore, mechanisms underlying the cell death and its potential use for treatment of skin cancer will be further studied.

SOS gene induction and possible mutagenic effects of freeze-drying in Escherichia coli and Salmonella typhimurium.

PubMed

Rosen, Rachel; Buchinger, Sebastian; Pfänder, Ramona; Pedhazur, Rami; Reifferscheid, Georg; Belkin, Shimshon

2016-11-01

We report the results of a study of the potential negative effects of the freeze-drying process, normally considered a benign means for long-term conservation of living cells and the golden standard in bacterial preservation. By monitoring gene induction using a whole-cell Escherichia coli bioreporter panel, in which diverse stress-responsive gene promoters are fused to luminescent or fluorescent reporting systems, we have demonstrated that DNA repair genes belonging to the SOS operon (recA, sulA, uvrA, umuD, and lexA) were induced upon resuscitation from the freeze-dried state, whereas other stress-responsive promoters such as grpE, katG, phoA, soxS, and sodA were not affected. This observation was confirmed by the UMU-chromotest (activation of the umuD gene promoter) in Salmonella typhimurium, as well as by real-time PCR analyses of selected E. coli SOS genes. We further show that a functional SOS operon is important in viability maintenance following resuscitation, but that at the same time, this repair system may introduce significantly higher mutation rates, comparable to those induced by high concentrations of a known mutagen. Our results also indicate that the entire freeze-drying process, rather than either freezing or drying separately, is instrumental in the induction of DNA damage.

Comparative molecular field analysis to derive pharmacophore maps for induction doses of intravenous anaesthetic agents.

PubMed

Sear, J W

2011-03-01

The present study examines the molecular basis of induction of anaesthesia by i.v. hypnotic agents using comparative molecular field analysis (CoMFA). ED(50) induction doses for 14 i.v. anaesthetics in human subjects (expressed as molar dose per kilogram body weight) were obtained from the literature. Immobilizing potency data for the same 14 agents (expressed as the EC(50) plasma free drug concentrations that abolish movement in response to a noxious stimulus in 50% patients) were taken from our previous publication. These data were used to form CoMFA models for the two aspects of anaesthetic activity. Molecular alignment was achieved by field-fit minimization techniques. The lead structure for both models was eltanolone. The final CoMFA model for the ED(50) induction dose was based on two latent variables, and explained 99.3% of the variance in observed activities. It showed good intrinsic predictability (cross-validated q(2)=0.849). The equivalent model for immobilizing activity was also based on two latent variables, with r(2)=0.988 and q(2)=0.852. Although there was a correlation between -log ED(50) and -log EC(50) (r(2)=0.779), comparison of the pharmacophore maps showed poor correlation for both electrostatic and steric regions when isocontours were constructed by linking lattice grid points, making the greatest 40% contributions; the relative contributions of electrostatic and steric interactions differing between the models (induction dose: 2.5:1; immobilizing activity 1.8:1). Comparison of two CoMFA activity models shows only small elements of commonality, suggesting that different molecular features may be responsible for these two properties of i.v. anaesthetics.

Myc Dynamically and Preferentially Relocates to a Transcription Factory Occupied by Igh

PubMed Central

Osborne, Cameron S; Chakalova, Lyubomira; Mitchell, Jennifer A; Horton, Alice; Wood, Andrew L; Bolland, Daniel J; Corcoran, Anne E; Fraser, Peter

2007-01-01

Transcription in mammalian nuclei is highly compartmentalized in RNA polymerase II-enriched nuclear foci known as transcription factories. Genes in cis and trans can share the same factory, suggesting that genes migrate to preassembled transcription sites. We used fluorescent in situ hybridization to investigate the dynamics of gene association with transcription factories during immediate early (IE) gene induction in mouse B lymphocytes. Here, we show that induction involves rapid gene relocation to transcription factories. Importantly, we find that the Myc proto-oncogene on Chromosome 15 is preferentially recruited to the same transcription factory as the highly transcribed Igh gene located on Chromosome 12. Myc and Igh are the most frequent translocation partners in plasmacytoma and Burkitt lymphoma. Our results show that transcriptional activation of IE genes involves rapid relocation to preassembled transcription factories. Furthermore, the data imply a direct link between the nonrandom interchromosomal organization of transcribed genes at transcription factories and the incidence of specific chromosomal translocations. PMID:17622196

Chiral symmetry breaking by spatial confinement in tactoidal droplets of lyotropic chromonic liquid crystals

PubMed Central

Tortora, Luana; Lavrentovich, Oleg D.

2011-01-01

In many colloidal systems, an orientationally ordered nematic (N) phase emerges from the isotropic (I) melt in the form of spindle-like birefringent tactoids. In cases studied so far, the tactoids always reveal a mirror-symmetric nonchiral structure, sometimes even when the building units are chiral. We report on chiral symmetry breaking in the nematic tactoids formed in molecularly nonchiral polymer-crowded aqueous solutions of low-molecular weight disodium cromoglycate. The parity is broken by twisted packing of self-assembled molecular aggregates within the tactoids as manifested by the observed optical activity. Fluorescent confocal microscopy reveals that the chiral N tactoids are located at the boundaries of cells. We explain the chirality induction as a replacement of energetically costly splay packing of the aggregates within the curved bipolar tactoidal shape with twisted packing. The effect represents a simple pathway of macroscopic chirality induction in an organic system with no molecular chirality, as the only requirements are orientational order and curved shape of confinement. PMID:21402929

Spatial distribution and historical trends of heavy metals in the sediments of petroleum producing regions of the Beibu Gulf, China.

PubMed

Yang, Jichao; Wang, Weiguo; Zhao, Mengwei; Chen, Bin; Dada, Olusegun A; Chu, Zhihui

2015-02-15

The concentrations of As, Sb, Hg, Pb, Cd, and Ba in the surface and core sediments of the oil and gas producing region of the Beibu Gulf were measured by Inductively Coupled Plasma Optical Emission Spectrometry (ICP-OES), Inductively Coupled Plasma Mass Spectrometry (ICP-MS) and Atomic Fluorescence Spectrometry (AFS), and the spatial distribution and historical trends of these elements are discussed. The results show that the concentrations of these elements are highest near the platforms. The results of Enrichment Factor (EF) and Potential Ecological Risk Index (PERI) also reveal significantly higher enrichment around the platforms, which imply that the offshore petroleum production was the cause of the unusual distribution and severe enrichment of these elements in the study area. The environment around the platforms was highly laden with toxic elements, thereby representing a very high ecological risk to the environment of the study area. Copyright © 2014 Elsevier Ltd. All rights reserved.

Development of a Plastic Embedding Method for Large-Volume and Fluorescent-Protein-Expressing Tissues

PubMed Central

Yang, Zhongqin; Hu, Bihe; Zhang, Yuhui; Luo, Qingming; Gong, Hui

2013-01-01

Fluorescent proteins serve as important biomarkers for visualizing both subcellular organelles in living cells and structural and functional details in large-volume tissues or organs. However, current techniques for plastic embedding are limited in their ability to preserve fluorescence while remaining suitable for micro-optical sectioning tomography of large-volume samples. In this study, we quantitatively evaluated the fluorescence preservation and penetration time of several commonly used resins in a Thy1-eYFP-H transgenic whole mouse brain, including glycol methacrylate (GMA), LR White, hydroxypropyl methacrylate (HPMA) and Unicryl. We found that HMPA embedding doubled the eYFP fluorescence intensity but required long durations of incubation for whole brain penetration. GMA, Unicryl and LR White each penetrated the brain rapidly but also led to variable quenching of eYFP fluorescence. Among the fast-penetrating resins, GMA preserved fluorescence better than LR White and Unicryl. We found that we could optimize the GMA formulation by reducing the polymerization temperature, removing 4-methoxyphenol and adjusting the pH of the resin solution to be alkaline. By optimizing the GMA formulation, we increased percentage of eYFP fluorescence preservation in GMA-embedded brains nearly two-fold. These results suggest that modified GMA is suitable for embedding large-volume tissues such as whole mouse brain and provide a novel approach for visualizing brain-wide networks. PMID:23577174

PDT dose dosimetry for Photofrin-mediated pleural photodynamic therapy (pPDT)

NASA Astrophysics Data System (ADS)

Ong, Yi Hong; Kim, Michele M.; Finlay, Jarod C.; Dimofte, Andreea; Singhal, Sunil; Glatstein, Eli; Cengel, Keith A.; Zhu, Timothy C.

2018-01-01

Photosensitizer fluorescence excited by photodynamic therapy (PDT) treatment light can be used to monitor the in vivo concentration of the photosensitizer and its photobleaching. The temporal integral of the product of in vivo photosensitizer concentration and light fluence is called PDT dose, which is an important dosimetry quantity for PDT. However, the detected photosensitizer fluorescence may be distorted by variations in the absorption and scattering of both excitation and fluorescence light in tissue. Therefore, correction of the measured fluorescence for distortion due to variable optical properties is required for absolute quantification of photosensitizer concentration. In this study, we have developed a four-channel PDT dose dosimetry system to simultaneously acquire light dosimetry and photosensitizer fluorescence data. We measured PDT dose at four sites in the pleural cavity during pleural PDT. We have determined an empirical optical property correction function using Monte Carlo simulations of fluorescence for a range of physiologically relevant tissue optical properties. Parameters of the optical property correction function for Photofrin fluorescence were determined experimentally using tissue-simulating phantoms. In vivo measurements of photosensitizer fluorescence showed negligible photobleaching of Photofrin during the PDT treatment, but large intra- and inter-patient heterogeneities of in vivo Photofrin concentration are observed. PDT doses delivered to 22 sites in the pleural cavity of 8 patients were different by 2.9 times intra-patient and 8.3 times inter-patient.

Elimination of autofluorescence background from fluorescence tissue images by use of time-gated detection and the AzaDiOxaTriAngulenium (ADOTA) fluorophore

PubMed Central

Rich, Ryan M.; Stankowska, Dorota L.; Maliwal, Badri P.; Sørensen, Thomas Just; Laursen, Bo W.; Krishnamoorthy, Raghu R.; Gryczynski, Zygmunt; Borejdo, Julian

2013-01-01

Sample autofluorescence (fluorescence of inherent components of tissue and fixative-induced fluorescence) is a significant problem in direct imaging of molecular processes in biological samples. A large variety of naturally occurring fluorescent components in tissue results in broad emission that overlaps the emission of typical fluorescent dyes used for tissue labeling. In addition, autofluorescence is characterized by complex fluorescence intensity decay composed of multiple components whose lifetimes range from sub-nanoseconds to a few nanoseconds. For these reasons, the real fluorescence signal of the probe is difficult to separate from the unwanted autofluorescence. Here we present a method for reducing the autofluorescence problem by utilizing an azadioxatriangulenium (ADOTA) dye with a fluorescence lifetime of approximately 15 ns, much longer than those of most of the components of autofluorescence. A probe with such a long lifetime enables us to use time-gated intensity imaging to separate the signal of the targeting dye from the autofluorescence. We have shown experimentally that by discarding photons detected within the first 20 ns of the excitation pulse, the signal-to-background ratio is improved fivefold. This time-gating eliminates over 96 % of autofluorescence. Analysis using a variable time-gate may enable quantitative determination of the bound probe without the contributions from the background. PMID:23254457

Chloro-Functionalized Photo-crosslinking BODIPY for Glutathione Sensing and Subcellular Trafficking.

PubMed

Murale, Dhiraj P; Hong, Seong Cheol; Haque, Md Mamunul; Lee, Jun-Seok

2018-05-18

Glutathione (GSH) is one of major antioxidants inside cells that regulates oxidoreduction homeostasis. Recently, there have been extensive efforts to visualize GSH in live cells, but most of the probes available today are simple detection sensors and do not provide details of cellular localization. A new fluorescent probe (pcBD2-Cl), which is cell permeable and selectively reacts with GSH in situ, has been developed. The in situ GSH-labeled probe (pcBD2-GSH) exhibited quenches fluorescence, but subsequent binding to cellular abundant glutathione S-transferase (GST) recovers the fluorescence intensity, which makes it possible to image the GSH-GST complex in live cells. Interactions between probe and GST were confirmed by means of photo-crosslinking under intact live-cell conditions. Interestingly, isomers of chloro-functionalized 4,4-difluoro-4-bora-3a,4a-diaza-s-indacene (BODIPY) compounds behaved very distinctively inside the cells. Following co-staining imaging with MitoTracker and mitochondria fractionation upon lipopolysaccharide-mediated reactive oxygen species induction experiments showed that pcBD2-GSH accumulated in mitochondria. This is the first example of a live-cell imaging probe to visualize translocation of GSH from the cytosol to mitochondria. © 2018 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

Fluorescence-based optimization of human bitter taste receptor expression in Saccharomyces cerevisiae

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

Sugawara, Taishi; Ito, Keisuke; Shiroishi, Mitsunori

2009-05-15

Human TAS2 receptors (hTAS2Rs) perceive bitter tastants, but few studies have explored the structure-function relationships of these receptors. In this paper, we report our trials on the large-scale preparations of hTAS2Rs for structural analysis. Twenty-five hTAS2Rs were expressed using a GFP-fusion yeast system in which the constructs and the culture conditions (e.g., the signal sequence, incubation time and temperature after induction) were optimized by measuring GFP fluorescence. After optimization, five hTAS2Rs (hTAS2R7, hTAS2R8, hTAS2R16, hTAS2R41, and hTAS2R48) were expressed at levels greater than 1 mg protein/L of culture, which is a preferable level for purification and crystallization. Among these fivemore » bitter taste receptors, hTAS2R41 exhibited the highest detergent solubilization efficiency of 87.1% in n-dodecyl-{beta}-D-maltopyranoside (DDM)/cholesteryl hemisuccinate (CHS). Fluorescence size-exclusion chromatography showed that hTAS2R41 exhibited monodispersity in DDM/CHS without aggregates, suggesting that hTAS2R41 is a good target for future crystallization trials.« less

Quantitative imaging of gold nanoparticle distribution in a tumor-bearing mouse using benchtop x-ray fluorescence computed tomography

PubMed Central

Manohar, Nivedh; Reynoso, Francisco J.; Diagaradjane, Parmeswaran; Krishnan, Sunil; Cho, Sang Hyun

2016-01-01

X-ray fluorescence computed tomography (XFCT) is a technique that can identify, quantify, and locate elements within objects by detecting x-ray fluorescence (characteristic x-rays) stimulated by an excitation source, typically derived from a synchrotron. However, the use of a synchrotron limits practicality and accessibility of XFCT for routine biomedical imaging applications. Therefore, we have developed the ability to perform XFCT on a benchtop setting with ordinary polychromatic x-ray sources. Here, we report our postmortem study that demonstrates the use of benchtop XFCT to accurately image the distribution of gold nanoparticles (GNPs) injected into a tumor-bearing mouse. The distribution of GNPs as determined by benchtop XFCT was validated using inductively coupled plasma mass spectrometry. This investigation shows drastically enhanced sensitivity and specificity of GNP detection and quantification with benchtop XFCT, up to two orders of magnitude better than conventional x-ray CT. The results also reaffirm the unique capabilities of benchtop XFCT for simultaneous determination of the spatial distribution and concentration of nonradioactive metallic probes, such as GNPs, within the context of small animal imaging. Overall, this investigation identifies a clear path toward in vivo molecular imaging using benchtop XFCT techniques in conjunction with GNPs and other metallic probes. PMID:26912068

USE OF PULSE-AMPLITUDE-MODULATED FLUORESCENCE TO ASSESS THE PHYSIOLOGICAL STATUS OF CLADOPHORA SP. ALONG A WATER QUALITY GRADIENT(1).

PubMed

Hiriart-Baer, Véronique P; Arciszewski, Tim J; Malkin, Sairah Y; Guildford, Stephanie J; Hecky, Robert E

2008-12-01

This study investigated the application of pulse-amplitude-modulated (PAM) fluorometry as a rapid assessment of benthic macroalgal physiological status. Maximum quantum efficiency (Fv /Fm ), dark-light induction curves, and rapid fluorescence light-response curves (RLC) were measured on the filamentous macroalgal Cladophora sp. from Lake Ontario on 5 d at 16 sites spanning a gradient of light and nutrient supply. For Cladophora sp. growing in situ, light limitation was assessed by comparing average daily irradiance with the light utilization efficiency parameter (α) derived from RLCs. In this study, there was a nonlinear relationship between Fv /Fm and the degree of P limitation in macroalgae. However, only light-saturated Cladophora sp. showed a significant positive linear relationship between Fv /Fm and P nutrient status. The absence of this relationship among light-limited algae indicates that their photosynthetic rate would be stimulated by increased water clarity, and not by increased P supply. PAM fluorescence measures were successfully able to identify light-saturated macroalgae and, among these, assess the degree to which they were nutrient limited. These results enable us to test hypotheses arising from numeric models predicting the impact of changes in light penetration and nutrient supply on benthic primary production. © 2008 Phycological Society of America.

Investigation of selective induction of breast cancer cells to death with treatment of plasma-activated medium

NASA Astrophysics Data System (ADS)

Hashizume, Hiroshi; Tanaka, Hiromasa; Nakamura, Kae; Kano, Hiroyuki; Ishikawa, Kenji; Kikkawa, Fumitaka; Mizuno, Masaaki; Hori, Masaru

2015-09-01

The applications of plasma in medicine have much attention. We previously showed that plasma-activated medium (PAM) induced glioblastoma cells to apoptosis. However, it has not been elucidated the selectivity of PAM in detail. In this study, we investigated the selective effect of PAM on the death of human breast normal and cancer cells, MCF10A and MCF7, respectively, and observed the selective death with fluorescent microscopy. For the investigation of cell viability with PAM treatment, we prepared various PAMs according to the strengths, and treated each of cells with PAMs. Week PAM treatment only decreased the viability of MCF7 cells, while strong PAM treatment significantly affected both viabilities of MCF7 and MCF10A cells. For the fluorescent observation, we prepared the mixture of MCF7 and fluorescent-probed MCF10A cells, and seeded them. After the treatment of PAMs, the images showed that only MCF7 cells damaged in the mixture with week PAM treatment. These results suggested that a specific range existed with the selective effect in the strength of PAM. This work was partly supported by a Grant-in-Aid for Scientific Research on Innovative Areas ``Plasma Medical Innovation'' Grant No. 24108002 and 24108008 from the Ministry of Education, Culture, Sports, Science and Technology of Japan.

The observation of mitochondrial movement and ATG5 position in Arabidopsis during the process of infection with virulent and avirulent P. syringae strains

NASA Astrophysics Data System (ADS)

Yang, Liu; Ma, Chao; Chen, Wen li

2012-03-01

Infection of plants with pathogens leads to programmed cell death (PCD) associated with the pathogen-triggered hypersensitive response (HR) during plant innate immunity. In this study, the effects of infection by virulent Pseudomonas syringae pv. tomato (Pst) DC3000 and strains harboring avirulence factors AvrRps4 on the induction of HR-PCD were compared. We used Arabidopsis thaliana plants as materials, which expressed green fluorescent protein labeled mitochondria (mito-GFP) and green fluorescent protein tagged ATG5 (ATG5-GFP), these GFP are instantaneous expression. We found both Pst DC3000 and Pst-avrRps4 could induce mitochondria to assemble, the effect of Pst DC3000 was more obvious. ATG5 was located in chloroplasts after infection with Pst DC3000 or Pst-avrRps4. Under the condition of Pst-avrRps4, the expression of ATG5 was stronger than Pst DC3000 treatment.

Study of Linear and Nonlinear Wave Excitation

NASA Astrophysics Data System (ADS)

Chu, Feng; Berumen, Jorge; Hood, Ryan; Mattingly, Sean; Skiff, Frederick

2013-10-01

We report an experimental study of externally excited low-frequency waves in a cylindrical, magnetized, singly-ionized Argon inductively-coupled gas discharge plasma that is weakly collisional. Wave excitation in the drift wave frequency range is accomplished by low-percentage amplitude modulation of the RF plasma source. Laser-induced fluorescence is adopted to study ion-density fluctuations in phase space. The laser is chopped to separate LIF from collisional fluorescence. A single negatively-biased Langmuir probe is used to detect ion-density fluctuations in the plasma. A ring array of Langmuir probes is also used to analyze the spatial and spectral structure of the excited waves. We apply coherent detection with respect to the wave frequency to obtain the ion distribution function associated with externally generated waves. Higher-order spectra are computed to evaluate the nonlinear coupling between fluctuations at various frequencies produced by the externally generated waves. Parametric decay of the waves is observed. This work is supported by U.S. DOE Grant No. DE-FG02-99ER54543.

Functional recombinant protein is present in the pre-induction phases of Pichia pastoris cultures when grown in bioreactors, but not shake-flasks.

PubMed

Bawa, Zharain; Routledge, Sarah J; Jamshad, Mohammed; Clare, Michelle; Sarkar, Debasmita; Dickerson, Ian; Ganzlin, Markus; Poyner, David R; Bill, Roslyn M

2014-09-04

Pichia pastoris is a widely-used host for recombinant protein production; expression is typically driven by methanol-inducible alcohol oxidase (AOX) promoters. Recently this system has become an important source of recombinant G protein-coupled receptors (GPCRs) for structural biology and drug discovery. The influence of diverse culture parameters (such as pH, dissolved oxygen concentration, medium composition, antifoam concentration and culture temperature) on productivity has been investigated for a wide range of recombinant proteins in P. pastoris. In contrast, the impact of the pre-induction phases on yield has not been as closely studied. In this study, we examined the pre-induction phases of P. pastoris bioreactor cultivations producing three different recombinant proteins: the GPCR, human A(2a) adenosine receptor (hA(2a)R), green fluorescent protein (GFP) and human calcitonin gene-related peptide receptor component protein (as a GFP fusion protein; hCGRP-RCP-GFP). Functional hA(2a)R was detected in the pre-induction phases of a 1 L bioreactor cultivation of glycerol-grown P. pastoris. In a separate experiment, a glycerol-grown P. pastoris strain secreted soluble GFP prior to methanol addition. When glucose, which has been shown to repress AOX expression, was the pre-induction carbon source, hA(2a)R and GFP were still produced in the pre-induction phases. Both hA(2a)R and GFP were also produced in methanol-free cultivations; functional protein yields were maintained or increased after depletion of the carbon source. Analysis of the pre-induction phases of 10 L pilot scale cultivations also demonstrated that pre-induction yields were at least maintained after methanol induction, even in the presence of cytotoxic concentrations of methanol. Additional bioreactor data for hCGRP-RCP-GFP and shake-flask data for GFP, horseradish peroxidase (HRP), the human tetraspanins hCD81 and CD82, and the tight-junction protein human claudin-1, demonstrated that bioreactor but not shake-flask cultivations exhibit recombinant protein production in the pre-induction phases of P. pastoris cultures. The production of recombinant hA(2a)R, GFP and hCGRP-RCP-GFP can be detected in bioreactor cultivations prior to methanol induction, while this is not the case for shake-flask cultivations of GFP, HRP, hCD81, hCD82 and human claudin-1. This confirms earlier suggestions of leaky expression from AOX promoters, which we report here for both glycerol- and glucose-grown cells in bioreactor cultivations. These findings suggest that the productivity of AOX-dependent bioprocesses is not solely dependent on induction by methanol. We conclude that in order to maximize total yields, pre-induction phase cultivation conditions should be optimized, and that increased specific productivity may result in decreased biomass yields.

Fluorescent Reporter Libraries as Useful Tools for Optimizing Microbial Cell Factories: A Review of the Current Methods and Applications

PubMed Central

Delvigne, Frank; Pêcheux, Hélène; Tarayre, Cédric

2015-01-01

The use of genetically encoded fluorescent reporters allows speeding up the initial optimization steps of microbial bioprocesses. These reporters can be used for determining the expression level of a particular promoter, not only the synthesis of a specific protein but also the content of intracellular metabolites. The level of protein/metabolite is thus proportional to a fluorescence signal. By this way, mean expression profiles of protein/metabolites can be determined non-invasively at a high-throughput rate, allowing the rapid identification of the best producers. Actually, different kinds of reporter systems are available, as well as specific cultivation devices allowing the on-line recording of the fluorescent signal. Cell-to-cell variability is another important phenomenon that can be integrated into the screening procedures for the selection of more efficient microbial cell factories. PMID:26442261

Fluorescence characterization of fractionated dissolved organic matter in the five tributaries of Poyang Lake, China.

PubMed

Yan, Caixia; Liu, Huihui; Sheng, Yanru; Huang, Xian; Nie, Minghua; Huang, Qi; Baalousha, Mohammed

2018-10-01

Characterization of natural colloids is the key to understand pollutant fate and transport in the environment. The present study investigates the relationship between size and fluorescence properties of colloidal organic matter (COM) from five tributaries of Poyang Lake. Colloids were size-fractionated using cross-flow ultrafiltration and their fluorescence properties were measured by three-dimensional excitation-emission matrix fluorescence spectroscopy (3D-EEM). Parallel factor analysis (PARAFAC) and/or Self-organizing map (SOM) were applied to assess fluorescence properties as proxy indicators for the different size of colloids. PARAFAC analysis identified four fluorescence components including three humic-like components (C1-C3) and a protein-like component (C4). These four fluorescence components, and in particular the protein-like component, are primarily present in <1 kDa phase. For the colloidal fractions (1-10 kDa, 10-100 kDa, and 100 kDa-0.7 μm), the majority of fluorophores are associated with the smallest size fraction. SOM analysis demonstrated that relatively high fluorescence intensity and aromaticity occur primarily in <1 kDa phase, followed by 1-10 kDa colloids. Coupling PARAFAC and SOM facilitate the visualization and interpretation of the relationship between colloidal size and fluorescence properties with fewer input variables, shorter running time, higher reliability, and nondestructive results. Fluorescence indices analysis reveals that the smallest colloidal fraction (1-10 kDa) was dominated by higher humified and less autochthonous COM. Copyright © 2018 Elsevier B.V. All rights reserved.

Doubly fed induction generator wind turbines with fuzzy controller: a survey.

PubMed

Sathiyanarayanan, J S; Kumar, A Senthil

2014-01-01

Wind energy is one of the extraordinary sources of renewable energy due to its clean character and free availability. With the increasing wind power penetration, the wind farms are directly influencing the power systems. The majority of wind farms are using variable speed wind turbines equipped with doubly fed induction generators (DFIG) due to their advantages over other wind turbine generators (WTGs). Therefore, the analysis of wind power dynamics with the DFIG wind turbines has become a very important research issue, especially during transient faults. This paper presents fuzzy logic control of doubly fed induction generator (DFIG) wind turbine in a sample power system. Fuzzy logic controller is applied to rotor side converter for active power control and voltage regulation of wind turbine.

Power Smoothing and MPPT for Grid-connected Wind Power Generation with Doubly Fed Induction Generator

NASA Astrophysics Data System (ADS)

Kai, Takaaki; Tanaka, Yuji; Kaneda, Hirotoshi; Kobayashi, Daichi; Tanaka, Akio

Recently, doubly fed induction generator (DFIG) and synchronous generator are mostly applied for wind power generation, and variable speed control and power factor control are executed for high efficiently for wind energy capture and high quality for power system voltage. In variable speed control, a wind speed or a generator speed is used for maximum power point tracking. However, performances of a wind generation power fluctuation due to wind speed variation have not yet investigated for those controls. The authors discuss power smoothing by those controls for the DFIG inter-connected to 6.6kV distribution line. The performances are verified using power system simulation software PSCAD/EMTDC for actual wind speed data and are examined from an approximate equation of wind generation power fluctuation for wind speed variation.

Study of Variable Frequency Induction Heating in Steel Making Process

NASA Astrophysics Data System (ADS)

Fukutani, Kazuhiko; Umetsu, Kenji; Itou, Takeo; Isobe, Takanori; Kitahara, Tadayuki; Shimada, Ryuichi

Induction heating technologies have been the standard technologies employed in steel making processes because they are clean, they have a high energy density, they are highly the controllable, etc. However, there is a problem in using them; in general, frequencies of the electric circuits have to be kept fixed to improve their power factors, and this constraint makes the processes inflexible. In order to overcome this problem, we have developed a new heating technique-variable frequency power supply with magnetic energy recovery switching. This technique helps us in improving the quality of steel products as well as the productivity. We have also performed numerical calculations and experiments to evaluate its effect on temperature distributions on heated steel plates. The obtained results indicate that the application of the technique in steel making processes would be advantageous.

The AAI index, the BIS index and end-tidal concentration during wash in and wash out of sevoflurane.

PubMed

Anderson, R E; Barr, G; Assareh, H; Jakobsson, J

2003-06-01

The bispectral index (BIS), auditory evoked potential index (AAI) and the end-tidal sevoflurane concentration were studied during induction and emergence in 10 ASA I-II patients. Both during 'wash-in' and 'wash-out' of sevoflurane, the AAI and BIS indices show huge variability and an overlap of indices between awake and not responding to command. This was the most pronounced during induction and the range of values was larger for the AAI index as compared with the BIS index. Mean (range) BIS was 85 (73-98) and 48 (10-83) awake and unconscious, respectively, and mean AAI index was 71 (43-99) and 21 (4-85), respectively. This study demonstrates the difficulties of using processed EEG variables in real time in a clinical situation of non-steady state pharmacodynamics.

Dexamethasone enhances agonist induction of tissue factor in monocytes but not in endothelial cells.

PubMed

Bottles, K D; Morrissey, J H

1993-06-01

Stimulation of monocytic cells by inflammatory agents such as bacterial lipopolysaccharide or tumour necrosis factor-alpha leads to the rapid and transient expression of tissue factor, the major cellular initiator of the extrinsic coagulation cascade in both haemostasis and tissue inflammation. In this study we investigated whether the synthetic anti-inflammatory glucocorticoid, dexamethasone, would inhibit agonist induction of tissue factor expression in both monocytes and endothelial cells. Surprisingly, dexamethasone significantly enhanced the induction of tissue factor expression by peripheral blood mononuclear cells and an established monocytic cell line, THP-1, in response to lipopolysaccharide or tumour necrosis factor-alpha. However, unlike monocytic cells, dexamethasone did not enhance agonist induction of tissue factor in endothelial cells. Synergistic enhancement of tissue factor expression by dexamethasone was also reflected in tissue factor mRNA levels in THP-1 cells, but was not the result of improved TF mRNA stability. Synergism between bacterial lipopolysaccharide and glucocorticoid in the induction of monocyte effector function is extremely unusual and may help to explain the variable outcome of glucocorticoid treatment of septic shock.

Smart Sensor for Online Detection of Multiple-Combined Faults in VSD-Fed Induction Motors

PubMed Central

Garcia-Ramirez, Armando G.; Osornio-Rios, Roque A.; Granados-Lieberman, David; Garcia-Perez, Arturo; Romero-Troncoso, Rene J.

2012-01-01

Induction motors fed through variable speed drives (VSD) are widely used in different industrial processes. Nowadays, the industry demands the integration of smart sensors to improve the fault detection in order to reduce cost, maintenance and power consumption. Induction motors can develop one or more faults at the same time that can be produce severe damages. The combined fault identification in induction motors is a demanding task, but it has been rarely considered in spite of being a common situation, because it is difficult to identify two or more faults simultaneously. This work presents a smart sensor for online detection of simple and multiple-combined faults in induction motors fed through a VSD in a wide frequency range covering low frequencies from 3 Hz and high frequencies up to 60 Hz based on a primary sensor being a commercially available current clamp or a hall-effect sensor. The proposed smart sensor implements a methodology based on the fast Fourier transform (FFT), RMS calculation and artificial neural networks (ANN), which are processed online using digital hardware signal processing based on field programmable gate array (FPGA).

Icing-Protection Requirements for Reciprocating-Engine Induction System

NASA Technical Reports Server (NTRS)

Coles, Willard D; Rollin, Vern G; Mulholland, Donald R

1950-01-01

Despite the development of relatively ice-free fuel-metering systems, the widespread use of alternate and heated-air intakes, and the use of alcohol for emergency de-icing, icing of aircraft-engine induction systems is a serious problem. Investigations have been made to study and to combat all phases of this icing problem. From these investigations, criterions for safe operation and for design of new induction systems have been established. The results were obtained from laboratory investigations of carburetor-supercharger combinations, wind-tunnel investigations of air scoops, multicylinder-engine studies, and flight investigations. Characteristics of three forms of ice, impact, throttling, and fuel evaporation were studied. The effects of several factors on the icing characteristics were also studied and included: (1) atmospheric conditions, (2) engine and air-scoop configurations, including light-airplane system, (3) type fuel used, and (4) operating variables, such as power condition, use of a manifold pressure regulator, mixture setting, carburetor heat, and water-alcohol injection. In addition, ice-detection methods were investigated and methods of preventing and removing induction-system ice were studied. Recommendations are given for design and operation with regard to induction-system design.

T-FLAP improvement for VOS Program

NASA Astrophysics Data System (ADS)

Marcelli, Marco; Piermattei, Viviana; Madonia, Alice; Mainardi, Umberto; Manzella, Giuseppe M. R.

2010-05-01

The operational oceanography has been engaged in the development of new acquisition, transmission and assimilation systems in order to have the widest possible coverage of real time informations, reflecting the guidelines of the World Meteorological Organization (WMO) and of the Intergovernmental Oceanographic Commission (IOC). Physical and biological processes of marine ecosystems have a high spatial and temporal variability, whose study is possible only through high resolution and synoptic observations. More than for the physical variables, the biological ones have to be observed in situ. Especially in the mid-high latitudes, a deep observation of the water column is needed, because of the typical distribution of phytoplankton's biomass (Mann and Lazier, 1991). In the last times an extensive use of XBT was performed in order to provide near real time analysis of the ocean temperature, but there is still a lack in the biomass estimation. T-FLAP technology (Temperature and Fluorescence LAunchable Probe - Marcelli et al. 2007) was designed to answer to the claim of a cost effective temperature and fluorescence autonomous probe, to be used on ships of opportunity for the Voluntary Observing Ship Program (VOS). During the last three years the probe was upgraded and improved both in materials and in measure sensitivity. New LEDs were mounted to increase the radiant power of the excitation source.. In order to evaluate the red filter efficiency to detect fluorescence chlorophyll a emission wavelengths, transmittance of new available filters in the range 682-685 nm was calculated. In vivo fluorescence spectra of each filter were also performed on samples from Chlorella sp. cultures, to assess the percentage of the fluorescence emission peak transmitted by the filters. A multiple system was designed and realized for a high accuracy dynamic calibration of the probes.

A step towards standardization: A method for end-point titer determination by fluorescence index of an automated microscope. End-point titer determination by fluorescence index.

PubMed

Carbone, Teresa; Gilio, Michele; Padula, Maria Carmela; Tramontano, Giuseppina; D'Angelo, Salvatore; Pafundi, Vito

2018-05-01

Indirect Immunofluorescence (IIF) is widely considered the Gold Standard for Antinuclear Antibody (ANA) screening. However, the high inter-reader variability remains the major disadvantage associated with ANA testing and the main reason for the increasing demand of the computer-aided immunofluorescence microscope. Previous studies proposed the quantification of the fluorescence intensity as an alternative for the classical end-point titer evaluation. However, the different distribution of bright/dark light linked to the nature of the self-antigen and its location in the cells result in different mean fluorescence intensities. The aim of the present study was to correlate Fluorescence Index (F.I.) with end-point titers for each well-defined ANA pattern. Routine serum samples were screened for ANA testing on HEp-2000 cells using Immuno Concepts Image Navigator System, and positive samples were serially diluted to assign the end-point titer. A comparison between F.I. and end-point titers related to 10 different staining patterns was made. According to our analysis, good technical performance of F.I. (97% sensitivity and 94% specificity) was found. A significant correlation between quantitative reading of F.I. and end-point titer groups was observed using Spearman's test and regression analysis. A conversion scale of F.I. in end-point titers for each recognized ANA-pattern was obtained. The Image Navigator offers the opportunity to improve worldwide harmonization of ANA test results. In particular, digital F.I. allows quantifying ANA titers by using just one sample dilution. It could represent a valuable support for the routine laboratory and an effective tool to reduce inter- and intra-laboratory variability. Copyright © 2018. Published by Elsevier B.V.

Variable-Speed Induction Motor Drives for Aircraft Environmental Control Compressors

NASA Technical Reports Server (NTRS)

Mildice, J. W.; Hansen, I. G.; Schreiner, K. E.; Roth, M. E.

1996-01-01

New, more-efficient designs for aircraft jet engines are not capable of supplying the large quantities of bleed air necessary to provide pressurization and air conditioning for the environmental control systems (ECS) of the next generation of large passenger aircraft. System analysis and engineering have determined that electrically-driven ECS can help to maintain the improved fuel efficiencies; and electronic controllers and induction motors are now being developed in a NASA/NPD SBIR Program to drive both types of ECS compressors. Previous variable-speed induction motor/controller system developments and publications have primarily focused on field-oriented control, with large transient reserve power, for maximum acceleration and optimum response in actuator and robotics systems. The application area addressed herein is characterized by slowly-changing inputs and outputs, small reserve power capability for acceleration, and optimization for maximum efficiency. This paper therefore focuses on the differences between this case and the optimum response case, and shows the development of this new motor/controller approach. It starts with the creation of a new set of controller requirements. In response to those requirements, new control algorithms are being developed and implemented in an embedded computer, which is integrated into the motor controller closed loop. Buffered logic outputs are used to drive the power switches in a resonant-technology, power processor/motor-controller, at switching/resonant frequencies high enough to support efficient high-frequency induction motor operation at speeds up to 50,000-RPA

[Cesarean after labor induction: Risk factors and prediction score].

PubMed

Branger, B; Dochez, V; Gervier, S; Winer, N

2018-05-01

The objective of the study is to determine the risk factors for caesarean section at the time of labor induction, to establish a prediction algorithm, to evaluate its relevance and to compare the results with observation. A retrospective study was carried out over a year at Nantes University Hospital with 941 cervical ripening and labor inductions (24.1%) terminated by 167 caesarean sections (17.8%). Within the cohort, a case-control study was conducted with 147 caesarean sections and 148 vaginal deliveries. A multivariate analysis was carried out with a logistic regression allowing the elaboration of an equation of prediction and an ROC curve and the confrontation between the prediction and the reality. In univariate analysis, six variables were significant: nulliparity, small size of the mother, history of scarried uterus, use of prostaglandins as a mode of induction, unfavorable Bishop score<6, variety of posterior release. In multivariate analysis, five variables were significant: nulliparity, maternal size, maternal BMI, scar uterus and Bishop score. The most predictive model corresponded to an area under the curve of 0.86 (0.82-0.90) with a correct prediction percentage ("well classified") of 67.6% for a caesarean section risk of 80%. The prediction criteria would make it possible to inform the woman and the couple about the potential risk of Caesarean section in urgency or to favor a planned Caesarean section or a low-lying attempt on more objective, repeatable and transposable arguments in a medical team. Copyright © 2018 Elsevier Masson SAS. All rights reserved.

Improving Performance and Operational Stability of Porcine Interferon-α Production by Pichia pastoris with Combinational Induction Strategy of Low Temperature and Methanol/Sorbitol Co-feeding.

PubMed

Gao, Min-Jie; Zhan, Xiao-Bei; Gao, Peng; Zhang, Xu; Dong, Shi-Juan; Li, Zhen; Shi, Zhong-Ping; Lin, Chi-Chung

2015-05-01

Various induction strategies were investigated for effective porcine interferon-α (pIFN-α) production by Pichia pastoris in a 10 L fermenter. We found that pIFN-α concentration could be significantly improved with the strategies of low-temperature induction or methanol/sorbitol co-feeding. On this basis, a combinational strategy of induction at lower temperature (20 °C) with methanol/sorbitol co-feeding has been proposed for improvement of pIFN-α production. The results reveal that maximal pIFN-α concentration and antiviral activity reach the highest level of 2.7 g/L and 1.8 × 10(7) IU/mg with the proposed induction strategy, about 1.3-2.1 folds higher than those obtained with other sub-optimal induction strategies. Metabolic analysis and online multi-variable measurement results indicate that energy metabolic enrichment is responsible for the performance enhancement of pIFN-α production, as a large amount of ATP could be simultaneously produced from both formaldehyde oxidation pathway in methanol metabolism and tricarboxylic acid (TCA) cycle in sorbitol metabolism. In addition, the proposed combinational induction strategy enables P. pastoris to be resistant to high methanol concentration (42 g/L), which conceivably occur associating with the error-prone methanol over-feeding. As a result, the proposed combinational induction strategy simultaneously increased the targeted protein concentration and operational stability leading to significant improvement of pIFN-α production.

[Effects of long time different negative pressures on osteogenic differentiation of rabbit bone mesenchymal stem cells].

PubMed

Zhao, Bowen; Zhang, Hongwei; Xu, Qiang; Ge, Quanhu; Li, Bolong; Peng, Xinyu; Wu, Xiangwei

2017-05-01

To investigate the effects of long time different negative pressures on osteogenic diffe-rentiation of rabbit bone mesenchymal stem cells (BMSCs). The rabbit BMSCs were isolated and cultured by density gradient centrifugation. Flow cytometry was used to analyze expression of surface markers. The third passage cells cultured under condition of osteogenic induction and under different negative pressure of 0 mm Hg (control group), 75 mm Hg (low negative pressure group), and 150 mm Hg (high negative pressure group) (1 mm Hg=0.133 kPa), and the negative pressure time was 30 min/h. Cell growth was observed under phase contrast microscopy, and the growth curve was drawn; alkaline phosphatase (ALP) activity was detected by ELISA after induced for 3, 7, and 14 days. The mRNA and protein expressions of collagen type I (COL-I) and osteocalcin (OC) in BMSCs were analyzed by real-time fluorescence quantitative PCR and Western blot. The cultured cells were identified as BMSCs by flow cytometry. The third passage BMSCs exhibited typical long shuttle and irregular shape. Cell proliferation was inhibited with the increase of negative pressure. After induced for 4 days, the cell number of high negative pressure group was significantly less than that in control group and low negative pressure group ( P <0.05), but there was no significant difference between the low negative pressure group and the control group ( P >0.05); at 5-7 days, the cell number showed significant difference between 3 groups ( P <0.05). The greater the negative pressure was, the greater the inhibition of cell proliferation was. There was no significant difference in ALP activity between groups at 3 days after induction ( P >0.05); the ALP activity showed significant difference ( P <0.05) between the high negative pressure group and the control group at 7 days after induction; and significant difference was found in the ALP activity between 3 groups at 14 days after induction ( P <0.05). The greater the negative pressure was, the higher the ALP activity was. Real-time fluorescence quantitative PCR and Western blot detection showed that the mRNA and protein expressions of COL-I and OC protein were significantly higher in low negative pressure group and high negative pressure group than control group ( P <0.05), and in the high negative pressure group than the low negative pressure group ( P <0.05). With the increase of the negative pressure, the osteogenic differentiation ability of BMSCs increases gradually, but the cell proliferation is inhibited.

Induction of Intracellular Reductive Stress with a Photoactivatable Phosphine Probe.

PubMed

Tirla, Alina; Rivera-Fuentes, Pablo

2018-04-25

Reductive stress is a condition present in cells that have an increased concentration of reducing species, and it has been associated with a number of pathologies, such as neurodegenerative diseases and cancer. The tools available to study reductive stress lack both in selectivity and specific targeting and some of these shortcomings can be addressed by using photoactivatable compounds. We developed a photoactivatable phosphonium probe, which upon irradiation releases a fluorescent molecule and a trialkyphosphine. The probes can permeate through the plasma membrane and the photoreleased phosphine can induce intracellular reductive stress as proven by the detection of protein aggregates.

Distribution of heavy metals in the coastal area of Abu Dhabi in the United Arab Emirates.

PubMed

Al Rashdi, Saeed; Arabi, Alya A; Howari, Fares M; Siad, Abdi

2015-08-15

Fifty-seven sediment samples were collected from Abu Dhabi coastal area, United Arab Emirates (UAE). The concentrations of heavy metals including antimony, arsenic, barium, cadmium, cobalt, copper, mercury, lead, molybdenum, nickel and zinc were obtained using Inductively Coupled Plasma-Mass Spectroscopy (ICP-MS) and X-ray fluorescence. Heavy metal contaminations in Abu Dhabi had increased since 2004. Nevertheless, the enrichment factors, geoaccumulation indices and the pollution load index of 0.3 showed no pollution with any of the measured metals except arsenic. Copyright © 2015 Elsevier Ltd. All rights reserved.

Influence of land use and land cover on the spatial variability of dissolved organic matter in multiple aquatic environments.

PubMed

Singh, Shatrughan; Dash, Padmanava; Silwal, Saurav; Feng, Gary; Adeli, Ardeshir; Moorhead, Robert J

2017-06-01

Water quality of lakes, estuaries, and coastal areas serves as an indicator of the overall health of aquatic ecosystems as well as the health of the terrestrial ecosystem that drains to the water body. Land use and land cover plays not only a significant role in controlling the quantity of the exported dissolved organic matter (DOM) but also influences the quality of DOM via various biogeochemical and biodegradation processes. We examined the characteristics and spatial distribution of DOM in five major lakes, in an estuary, and in the coastal waters of the Mississippi, USA, and investigated the influence of the land use and land cover of their watersheds on the DOM composition. We employed absorption and fluorescence spectroscopy including excitation-emission matrix (EEM) combined with parallel factor (PARAFAC) analysis modeling techniques to determine optical properties of DOM and its characteristics in this study. We developed a site-specific PARAFAC model to evaluate DOM characteristics resulting in five diverse DOM compositions that included two terrestrial humic-like (C1 and C3), two microbial humic-like (C2 and C5), and one protein-like (C4) DOM. Our results showed elevated fluorescence levels of microbial humic-like or protein-like DOM in the lakes and coastal waters, while the estuarine waters showed relatively high fluorescence levels of terrestrial humic-like DOM. The results also showed that percent forest and wetland coverage explained 68 and 82% variability, respectively, in terrestrial humic-like DOM exports, while 87% variability in microbially derived humiclike DOM was explained by percent agricultural lands. Strong correlations between microbial humic-like DOM and fluorescence-derived DOM indices such as biological index (BIX) and fluorescence index (FI) indicated autochthonous characteristics in the lakes, while the estuary showed largely allochthonous DOM of terrestrial origin. We also observed higher concentrations of total dissolved phosphorous (TDP) and ammonium nitrogen (NH 4 -N) in coastal waters potentially due to photodegradation of refractory DOM derived from the sediment-bound organic matter in the coastal wetlands. This study highlights the relationships between the DOM compositions in the water and the land use and land cover in the watershed. The spatial variability of DOM in three different types of aquatic environments enhances the understanding of the role of land use and land cover in carbon cycling through export of organic matter to the aquatic ecosystems..

Transplantation of NSC-derived cholinergic neuron-like cells improves cognitive function in APP/PS1 transgenic mice.

PubMed

Gu, G; Zhang, W; Li, M; Ni, J; Wang, P

2015-04-16

The ability to selectively control the differentiation of neural stem cells (NSCs) into cholinergic neurons in vivo would be an important step toward cell replacement therapy. First, green fluorescent protein (GFP)-NSCs were induced to differentiate into cholinergic neuron-like cells (CNLs) with retinoic acid (RA) pre-induction followed by nerve growth factor (NGF) induction. Then, these CNLs were transplanted into bilateral hippocampus of APP/PS1 transgenic mice. Behavioral parameters showed by Morris water maze (MWM) tests and the percentages of GFP-labeled cholinergic neurons of CNL transplanted mice were compared with those of controls. Brain levels of choline acetyltransferase (ChAT) mRNA and proteins were analyzed by quantitative real-time PCR and Western blotting, ChAT activity and acetylcholine (ACh) concentration were also evaluated by ChAT activity and ACh concentration assay kits. Immunofluorescence analysis showed that 80.3±1.5% NSCs differentiated into CNLs after RA pre-induction followed by NGF induction in vitro. Three months after transplantation, 82.4±6.3% CNLs differentiated into cholinergic neurons in vivo. APP/PS1 mice transplanted with CNLs showed a significant improvement in learning and memory ability compared with control groups at different time points. Furthermore, CNLs transplantation dramatically increased in the expressions of ChAT mRNA and protein, as well ChAT activity and ACh concentration in APP/PS1 mice. Our findings support the prospect of using NSC-derived CNLs in developing therapies for Alzheimer's disease (AD). Copyright © 2015 IBRO. Published by Elsevier Ltd. All rights reserved.

Comparison of several analytical methods for the determination of tin in geochemical samples as a function of tin speciation

USGS Publications Warehouse

Kane, J.S.; Evans, J.R.; Jackson, J.C.

1989-01-01

Accurate and precise determinations of tin in geological materials are needed for fundamental studies of tin geochemistry, and for tin prospecting purposes. Achieving the required accuracy is difficult because of the different matrices in which Sn can occur (i.e. sulfides, silicates and cassiterite), and because of the variability of literature values for Sn concentrations in geochemical reference materials. We have evaluated three methods for the analysis of samples for Sn concentration: graphite furnace atomic absorption spectrometry (HGA-AAS) following iodide extraction, inductively coupled plasma atomic emission spectrometry (ICP-OES), and energy-dispersive X-ray fluorescence (EDXRF) spectrometry. Two of these methods (HGA-AAS and ICP-OES) required sample decomposition either by acid digestion or fusion, while the third (EDXRF) was performed directly on the powdered sample. Analytical details of all three methods, their potential errors, and the steps necessary to correct these errors were investigated. Results showed that similar accuracy was achieved from all methods for unmineralized samples, which contain no known Sn-bearing phase. For mineralized samples, which contain Sn-bearing minerals, either cassiterite or stannous sulfides, only EDXRF and fusion ICP-OES methods provided acceptable accuracy. This summary of our study provides information which helps to assure correct interpretation of data bases for underlying geochemical processes, regardless of method of data collection and its inherent limitations. ?? 1989.

Discrete wavelet transform and energy eigen value for rotor bars fault detection in variable speed field-oriented control of induction motor drive.

PubMed

Ameid, Tarek; Menacer, Arezki; Talhaoui, Hicham; Azzoug, Youness

2018-05-03

This paper presents a methodology for the broken rotor bars fault detection is considered when the rotor speed varies continuously and the induction machine is controlled by Field-Oriented Control (FOC). The rotor fault detection is obtained by analyzing a several mechanical and electrical quantities (i.e., rotor speed, stator phase current and output signal of the speed regulator) by the Discrete Wavelet Transform (DWT) in variable speed drives. The severity of the fault is obtained by stored energy calculation for active power signal. Hence, it can be a useful solution as fault indicator. The FOC is implemented in order to preserve a good performance speed control; to compensate the broken rotor bars effect in the mechanical speed and to ensure the operation continuity and to investigate the fault effect in the variable speed. The effectiveness of the technique is evaluated in simulation and in a real-time implementation by using Matlab/Simulink with the real-time interface (RTI) based on dSpace 1104 board. Copyright © 2018. Published by Elsevier Ltd.

Fluorescence self-quenching assay for the detection of target collagen sequences using a short probe peptide.

PubMed

Nian, Linge; Hu, Yue; Fu, Caihong; Song, Chen; Wang, Jie; Xiao, Jianxi

2018-01-01

The development of novel assays to detect collagen fragments is of utmost importance for diagnostic, prognostic and therapeutic decisions in various collagen-related diseases, and one essential question is to discover probe peptides that can specifically recognize target collagen sequences. Herein we have developed the fluorescence self-quenching assay as a convenient tool to screen the capability of a series of fluorescent probe peptides of variable lengths to bind with target collagen peptides. We have revealed that the targeting ability of probe peptides is length-dependent, and have discovered a relatively short probe peptide FAM-G(POG) 8 capable to identify the target peptide. We have further demonstrated that fluorescence self-quenching assay together with this short probe peptide can be applied to specifically detect the desired collagen fragment in complex biological media. Fluorescence self-quenching assay provides a powerful new tool to discover effective peptides for the recognition of collagen biomarkers, and it may have great potential to identify probe peptides for various protein biomarkers involved in pathological conditions. Copyright © 2017 Elsevier B.V. All rights reserved.

Reducing the background fluorescence in mice receiving fluorophore/inhibitor DNA duplexes.

PubMed

Liang, Minmin; Liu, Xinrong; Liu, Guozheng; Dou, Shuping; Cheng, Dengfeng; Liu, Yuxia; Rusckowski, Mary; Hnatowich, Donald J

2011-02-07

In principle, a DNA duplex consisting of an antisense fluorophore-conjugated major strand hybridized to a shorter complementary inhibitor-conjugated minor strand should provide fluorescence only in the tumor after intravenous administration if designed to remain intact except in the presence in tumor of its mRNA target. While we have obtained impressive tumor images in mice using this approach, there remains some background fluorescence. In this study, tissue homogenates of selected mouse organs were incubated with a test duplex and the kinetics of duplex dissociation in normal tissues were measured. In this manner we were able to identify the liver as the likely major source responsible for the duplex dissociation providing this fluorescence background. Thereafter liver homogenates were used to screen a series of duplex candidates with variable-length minor strands, and dissociation was measured by gel electrophoresis. The selected fluorophore/inhibitor duplex with improved stability displayed an insignificant (P > 0.05) background fluorescence after administration to SKH-1 normal mice and apparently without affecting target mRNA binding in vitro in cell culture or in vivo in tumor bearing mice.

Thermotolerance and Photosystem II Behaviour in Co-occuring Temperate Tree Species Exposed to Short-term Extreme Heat Waves

NASA Astrophysics Data System (ADS)

Guha, A.; Warren, J.; Cummings, C.; Han, J.

2017-12-01

Thermal stress can induce irreversible photodamage with longer consequences for plant metabolism. We focused on photosystem II (PSII) behaviour to understand how this complex responds in different co-occuring temperate trees exposed to short-term extreme heat waves. The study was designed for understanding complex heat tolerance mechanisms in trees. During manipulative heat-wave experiments, we monitored instantaneous PSII performance and tracked both transient and chronic PSII damages using chlorophyll a fluorescence characteristics. Fluorescence signals were used to simulate PSII bioenergetic processes. The light (Fv'/Fm') and dark-adapted (Fv/Fm) fluorescence traits including fast induction kinetics (OJIP), electron transport rate, PSII operating efficiency and quenching capacities were significantly affected by the heat treatments. Loss in PSII efficiency was more apparent in species like black cottonwood, yellow poplar, walnuts and conifers, whereas oaks maintained relatively better PSII functions. The post-heat recovery of Fv/Fm varied across the studied species showing differential carry over effects. PSII down-regulation was one of dominant factors for the loss in operational photosynthesis during extreme heat wave events. Both light and dark-adapted fluorescence characteristics showed loss in photo-regulatory functions and photodamage. Some resilient species showed rapid recovery from transient PSII damage, whereas fingerprints of chronic PSII damage were observed in susceptibles. Thresholds for Fv/Fm and non-photochemical quenching were identified for the studied species. PSII malfunctioning was largely associated with the observed photosynthetic down-regulation during heat wave treatments, however, its physiological recovery should be a key factor to determine species resilience to short-term extreme heat wave events.

Flow cytometric detection of micronuclei by combined staining of DNA and membranes

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

Wessels, J.M.; Nuesse, M.

1995-03-01

A new staining method is presented for flow cytometric measurement of micronuclei (MN) in cell cultures and human lymphocytes using membrane-specific fluorescent dyes in addition to DNA staining. Several combinations of fluorescent membrane and DNA dyes were studied for a better discrimination of MN from debris in a suspension of nuclei and micronuclei. For staining of membranes, the lipophilic dyes 2-hydroxyethyl-7,12,17-tris(methoxyethyl)porphycene (HEPn) and 1,6-diphenyl-1,3,5-hexatriene (DPH) were used in combination with ethidium bromide (EB), proflavine (PF), and Hoechst 33258 (HO). Due to their spectral properties, HO or EB combined with HEPn were not as suitable for the discrimination of MN frommore » debris as was HEPn in combination with PF. With HEPn in combination with PF, however, additional noise was found at low fluorescence intensities, probably due to free fluorescent dye molecules in the solution. The optimal simultaneous staining of membranes and DNA was obtained using a combination of DPH and EB. The induction of MN in Chinese hamster and mouse NIH-3T3 cells by UV-B illumination was studied with this new staining technique. UV-B illumination (280-360 nm) induced MN in both cell lines. Chinese hamster cells were found to be more sensitive to these wavelengths. Illumination with wavelengths above 360 nm did not induce MN in either cell line. The results obtained from human lymphocytes using the combination of EB or DPH were comparable to the results obtained with the combination of EB and HO. 23 refs., 7 figs.« less

Photoinduction of cyclosis-mediated interactions between distant chloroplasts.

PubMed

Bulychev, Alexander A; Komarova, Anna V

2015-01-01

Communications between chloroplasts and other organelles based on the exchange of metabolites, including redox active substances, are recognized as a part of intracellular regulation, chlororespiration, and defense against oxidative stress. Similar communications may operate between spatially distant chloroplasts in large cells where photosynthetic and respiratory activities are distributed unevenly under fluctuating patterned illumination. Microfluorometry of chlorophyll fluorescence in vivo in internodal cells of the alga Chara corallina revealed that a 30-s pulse of localized light induces a transient increase (~25%) in F' fluorescence of remote cell parts exposed to dim background light at a 1.5-mm distance on the downstream side from the illuminated spot in the plane of unilateral cytoplasmic streaming but has no effect on F' at equal distance on the upstream side. An abrupt arrest of cytoplasmic streaming for about 30s by triggering the action potential extended either the ascending or descending fronts of the F' fluorescence response, depending on the exact moment of streaming cessation. The response of F' fluorescence to localized illumination of a distant cell region was absent in dark-adapted internodes, when the localized light was applied within the first minute after switching on continuous background illumination of the whole cell, but it appeared in full after longer exposures to continuous background light. These results and the elimination of the F' response by methyl viologen known to redirect electron transport pathways beyond photosystem I indicate the importance of photosynthetic induction and the stromal redox state for long-distance communications of chloroplasts in vivo. Copyright © 2015 Elsevier B.V. All rights reserved.

Effect of Pre-treatment Nutritional Status, Folate and Vitamin B12 Levels on Induction Chemotherapy in Children with Acute Lymphoblastic Leukemia.

PubMed

Tandon, Sneha; Moulik, Nirmalya Roy; Kumar, Archana; Mahdi, Abbas Ali; Kumar, Ashutosh

2015-05-01

To evaluate pre-treatment undernutrition, and folate and B12 deficiency in children with acute lymphoblastic leukemia, and their correlation with complications and outcome of induction chemotherapy. Observational study. Tertiary care teaching hospital in Northern India. 50 children with acute lymphoblastic leukemia. Children were assessed for nutritional status (Weight for age Z-score, serum albumin, folate and B12) at presentation, and were followed-up during induction for bone marrow response, counts and outcome. Folate and B12 were repeated twice at monthly intervals after induction. Univariate and multivariate analyses were done to determine the association of nutritional parameters with the outcome variables. Baseline undernutrition was observed in 66%, hypo-albuminemia in 32.6%, folate deficiency in 41.3% and B12 deficiency in 36.9% of included children. Significant decline in folate levels was noted on serial assays during chemotherapy (P=0.001). Folate deficient children had higher risk for delayed marrow recovery and counts on day 14 (P=0.007 and P=0.001). Hypoalbuminemia (P=0.04), B12 deficiency (P=0.001) and folate (P=0.03) deficiency were associated with toxic deaths during induction. Baseline nutritional deficiencies negatively influence the outcome and occurrence of complications during induction chemotherapy in children with acute lymphoblastic leukemia.

Linear phase compressive filter

DOEpatents

McEwan, Thomas E.

1995-01-01

A phase linear filter for soliton suppression is in the form of a laddered series of stages of non-commensurate low pass filters with each low pass filter having a series coupled inductance (L) and a reverse biased, voltage dependent varactor diode, to ground which acts as a variable capacitance (C). L and C values are set to levels which correspond to a linear or conventional phase linear filter. Inductance is mapped directly from that of an equivalent nonlinear transmission line and capacitance is mapped from the linear case using a large signal equivalent of a nonlinear transmission line.

The 2013 FLEX-US Airborne Campaign at the Parker Tract Loblolly Pine Plantation in North Carolina, USA

NASA Technical Reports Server (NTRS)

Middleton, Elizabeth M.; Rascher, Uwe; Corp, Lawrence A.; Huemmrich, K. Fred; Cook, Bruce D.; Noormets, Asko; Schickling, Anke; Pinto, Francisco; Alonso, Luis; Damm, Alexander;

Control of fluorescence in quantum emitter and metallic nanoshell hybrids for medical applications

NASA Astrophysics Data System (ADS)

Singh, Mahi R.; Guo, Jiaohan; J. Cid, José M.; De Hoyos Martinez, Jesús E.

2017-03-01

We study the light emission from a quantum emitter and double metallic nanoshell hybrid systems. Quantum emitters act as local sources which transmit their light efficiently due to a double nanoshell near field. The double nanoshell consists of a dielectric core and two outer nanoshells. The first nanoshell is made of a metal, and the second spacer nanoshell is made of a dielectric material or human serum albumin. We have calculated the fluorescence emission for a quantum emitter-double nanoshell hybrid when it is injected in an animal or a human body. Surface plasmon polariton resonances in the double nanoshell are calculated using Maxwell's equations in the quasi-static approximation, and the fluorescence emission is evaluated using the density matrix method in the presence of dipole-dipole interactions. We have compared our theory with two fluorescence experiments in hybrid systems in which the quantum emitter is Indocyanine Green or infrared fluorescent molecules. The outer spacer nanoshell of double metallic nanoshells consists of silica and human serum albumin with variable thicknesses. Our theory explains the enhancement of fluorescence spectra in both experiments. We find that the thickness of the spacer nanoshell layer increases the enhancement when the fluorescence decreases. The enhancement of the fluorescence depends on the type of quantum emitter, spacer layer, and double nanoshell. We also found that the peak of the fluorescence spectrum can be shifted by changing the shape and the size of the nanoshell. The fluorescence spectra can be switched from one peak to two peaks by removing the degeneracy of excitonic states in the quantum emitter. Hence, using these properties, one can use these hybrids as sensing and switching devices for applications in medicine.

Estimating the leaf nitrogen content of paddy rice by using the combined reflectance and laser-induced fluorescence spectra.

PubMed

Yang, Jian; Du, Lin; Sun, Jia; Zhang, Zhenbing; Chen, Biwu; Shi, Shuo; Gong, Wei; Song, Shalei

2016-08-22

Paddy rice is one of the most important crops in China, and leaf nitrogen content (LNC) serves as a significant indictor for monitoring crop status. A reliable method is needed for precise and fast quantification of LNC. Laser-induced fluorescence (LIF) technology and reflectance spectra of crops are widely used to monitor leaf biochemical content. However, comparison between the fluorescence and reflectance spectra has been rarely investigated in the monitoring of LNC. In this study, the performance of the fluorescence and reflectance spectra for LNC estimation was discussed based on principal component analysis (PCA) and back-propagation neural network (BPNN). The combination of fluorescence and reflectance spectra was also proposed to monitor paddy rice LNC. The fluorescence and reflectance spectra exhibited a high degree of multi-collinearity. About 95.38%, and 97.76% of the total variance included in the spectra were efficiently extracted by using the first three PCs in PCA. The BPNN was implemented for LNC prediction based on new variables calculated using PCA. The experimental results demonstrated that the fluorescence spectra (R² = 0.810, 0.804 for 2014 and 2015, respectively) are superior to the reflectance spectra (R² = 0.721, 0.671 for 2014 and 2015, respectively) for estimating LNC based on the PCA-BPNN model. The proposed combination of fluorescence and reflectance spectra can greatly improve the accuracy of LNC estimation (R² = 0.912, 0.890 for 2014 and 2015, respectively).

Concentrations and characteristics of organic carbon in surface water in Arizona: Influence of urbanization

USGS Publications Warehouse

Westerhoff, P.; Anning, D.

2000-01-01

Dissolved (DOC) and total (TOC) organic carbon concentrations and compositions were studied for several river systems in Arizona, USA. DOC composition was characterized by ultraviolet and visible absorption and fluorescence emission (excitation wavelength of 370 nm) spectra characteristics. Ephemeral sites had the highest DOC concentrations, and unregulated perennial sites had lower concentrations than unregulated intermittent sites, regulated sites, and sites downstream from wastewater-treatment plants (p < 0.05). Reservoir outflows and wastewater-treatment plant effluent were higher in DOC concentration (p < 0.05) and exhibited less variability in concentration than inflows to the reservoirs. Specific ultraviolet absorbance values at 254 nm were typically less than 2 m-1(milligram DOC per liter)-1 and lower than values found in most temperate-region rivers, but specific ultraviolet absorbance values increased during runoff events. Fluorescence measurements indicated that DOC in desert streams typically exhibit characteristics of autochthonous sources; however, DOC in unregulated upland rivers and desert streams experienced sudden shifts from autochthonous to allochthonous sources during runoff events. The urban water system (reservoir systems and wastewater-treatment plants) was found to affect temporal variability in DOC concentration and composition. (C) 2000 Elsevier Science B.V.Dissolved (DOC) and total (TOC) organic carbon concentrations and compositions were studied for several river systems in Arizona, USA. DOC composition was characterized by ultraviolet and visible absorption and fluorescence emission (excitation wavelength of 370 nm) spectra characteristics. Ephemeral sites had the highest DOC concentrations, and unregulated perennial sites had lower concentrations than unregulated intermittent sites, regulated sites, and sites downstream from wastewater-treatment plants (p<0.05). Reservoir outflows and wastewater-treatment plant effluent were higher in DOC concentration (p<0.05) and exhibited less variability in concentration than inflows to the reservoirs. Specific ultraviolet absorbance values at 254 nm were typically less than 2 m-1(milligram DOC per liter)-1 and lower than values found in most temperate-region rivers, but specific ultraviolet absorbance values increased during runoff events. Fluorescence measurements indicated that DOC in desert streams typically exhibit characteristics of autochthonous sources; however, DOC in unregulated upland rivers and desert streams experienced sudden shifts from autochthonous to allochthonous sources during runoff events. The urban water system (reservoir systems and wastewater-treatment plants) was found to affect temporal variability in DOC concentration and composition.The influence of urbanization, becoming increasingly common in arid regions, on dissolved organic carbon (DOC) concentrations in surface water resources was studied. DOC concentration and composition, seasonal watershed runoff events, streamflow variations, water management practices, and urban infrastructure in several Arizona watersheds were monitored. Ephemeral sites had the highest DOC levels, and unregulated perennial sites and lower concentrations than unregulated intermittent sites, regulated sites, and sites downstream from wastewater treatment plants. Reservoir outflows and wastewater treatment plant effluent had higher and less variable DOC concentrations than inflows to reservoirs. UV absorbance values, fluorescence measurements, and other indicators suggest that urban water systems (reservoirs and wastewater treatment plants) affect temporal variability in DOC concentration and composition.

Apparatus for Teaching Physics.

ERIC Educational Resources Information Center

Gottlieb, Herbert H., Ed.

1981-01-01

Describes: (1) a variable inductor suitable for an inductance-capacitance bridge consisting of a fixed cylindrical solenoid and a moveable solenoid; (2) long-range apparatus for demonstrating falling bodies; and (3) an apparatus using two lasers to demonstrate ray optics. (SK)

Doubly Fed Induction Generator Wind Turbines with Fuzzy Controller: A Survey

PubMed Central

Sathiyanarayanan, J. S.; Senthil Kumar, A.

2014-01-01

Wind energy is one of the extraordinary sources of renewable energy due to its clean character and free availability. With the increasing wind power penetration, the wind farms are directly influencing the power systems. The majority of wind farms are using variable speed wind turbines equipped with doubly fed induction generators (DFIG) due to their advantages over other wind turbine generators (WTGs). Therefore, the analysis of wind power dynamics with the DFIG wind turbines has become a very important research issue, especially during transient faults. This paper presents fuzzy logic control of doubly fed induction generator (DFIG) wind turbine in a sample power system. Fuzzy logic controller is applied to rotor side converter for active power control and voltage regulation of wind turbine. PMID:25028677

Strain-specific induction of experimental autoimmune prostatitis (EAP) in mice.

PubMed

Jackson, Christopher M; Flies, Dallas B; Mosse, Claudio A; Parwani, Anil; Hipkiss, Edward L; Drake, Charles G

2013-05-01

Prostatitis, a clinical syndrome characterized by pelvic pain and inflammation, is common in adult males. Although several induced and spontaneous murine models of prostatitis have been explored, the role of genetic background on induction has not been well-defined. Using a standard methodology for the induction of experimental autoimmune prostatitis (EAP), we investigated both acute and chronic inflammation on several murine genetic backgrounds. In our colony, nonobese diabetic (NOD) mice evinced spontaneous prostatitis that was not augmented by immunization with rat prostate extract (RPE). In contrast, the standard laboratory strain Balb/c developed chronic inflammation in response to RPE immunization. Development of EAP in other strains was variable. These data suggest that Balb/c mice injected with RPE may provide a useful model for chronic prostatic inflammation. Copyright © 2012 Wiley Periodicals, Inc.

Strain-Specific Induction of Experimental Autoimmune Prostatitis (EAP) in Mice

PubMed Central

Jackson, Christopher M.; Flies, Dallas B.; Mosse, Claudio A.; Parwani, Anil; Hipkiss, Edward L.; Drake, Charles G.

2013-01-01

BACKGROUND Prostatitis, a clinical syndrome characterized by pelvic pain and inflammation, is common in adult males. Although several induced and spontaneous murine models of prostatitis have been explored, the role of genetic background on induction has not been well-defined. METHODS Using a standard methodology for the induction of experimental autoimmune prostatitis (EAP), we investigated both acute and chronic inflammation on several murine genetic backgrounds. RESULTS In our colony, nonobese diabetic (NOD) mice evinced spontaneous prostatitis that was not augmented by immunization with rat prostate extract (RPE). In contrast, the standard laboratory strain Balb/c developed chronic inflammation in response to RPE immunization. Development of EAP in other strains was variable. CONCLUSIONS These data suggest that Balb/c mice injected with RPE may provide a useful model for chronic prostatic inflammation. PMID:23129407

Influence of Eccentricity and Angular Velocity on Force Effects on Rotor of Magnetorheological Damper

NASA Astrophysics Data System (ADS)

Šedivý, Dominik; Ferfecki, Petr; Fialová, Simona

2018-06-01

This article presents the evaluation of force effects on squeeze film damper rotor. The rotor is placed eccentrically and its motion is translate-circular. The amplitude of rotor motion is smaller than its initial eccentricity. The force effects are calculated from pressure and viscous forces which were measured by using computational modeling. Damper was filled with magnetorheological fluid. Viscosity of this non-Newtonian fluid is given using Bingham rheology model. Yield stress is not constant and it is a function of magnetic induction which is described by many variables. The most important variables of magnetic induction are electric current and gap width between rotor and stator. The simulations were made in finite volume method based solver. The motion of the inner ring of squeeze film damper was carried out by dynamic mesh. Numerical solution was solved for five different initial eccentricities and angular velocities of rotor motion.

Force effects on rotor of squeeze film damper using Newtonian and non-Newtonian fluid

NASA Astrophysics Data System (ADS)

Dominik, Šedivý; Petr, Ferfecki; Simona, Fialová

2017-09-01

This article presents the evaluation of force effects on rotor of squeeze film damper. Rotor is eccentric placed and its motion is translate-circular. The amplitude of rotor motion is smaller than its initial eccentricity. The force effects are calculated from pressure and viscous forces which were gained by using computational modeling. Two types of fluid were considered as filling of damper. First type of fluid is Newtonian (has constant viscosity) and second type is magnetorheological fluid (does not have constant viscosity). Viscosity of non-Newtonian fluid is given using Bingham rheology model. Yield stress is a function of magnetic induction which is described by many variables. The most important variables of magnetic induction are electric current and gap width which is between rotor and stator. Comparison of application two given types of fluids is shown in results.

Can valproic acid be an inducer of clozapine metabolism?

PubMed Central

Diaz, Francisco J.; Eap, Chin B.; Ansermot, Nicolas; Crettol, Severine; Spina, Edoardo; de Leon, Jose

2014-01-01

Introduction Prior clozapine studies indicated no effects, mild inhibition or induction of valproic acid (VPA) on clozapine metabolism. The hypotheses that 1) VPA is a net inducer of clozapine metabolism, and 2) smoking modifies this inductive effect were tested in a therapeutic drug monitoring study. Methods After excluding strong inhibitors and inducers, 353 steady-state total clozapine (clozapine plus norclozapine) concentrations provided by 151 patients were analyzed using a random intercept linear model. Results VPA appeared to be an inducer of clozapine metabolism since total plasma clozapine concentrations in subjects taking VPA were significantly lower (27% lower; 95% confidence interval, 14% to 39%) after controlling for confounding variables including smoking (35% lower, 28% to 56%). Discussion Prospective studies are needed to definitively establish that VPA may 1) be an inducer of clozapine metabolism when induction prevails over competitive inhibition, and 2) be an inducer even in smokers who are under the influence of smoking inductive effects on clozapine metabolism. PMID:24764199

Laser-induced fluorescence studies of premalignant and benign lesions in the female genital tract

NASA Astrophysics Data System (ADS)

af Klinteberg, Claes; Wang, Ingrid; Lindquist, Charlotta; Vaitkuviene, Aurelija; Svanberg, Katarina

1997-12-01

Laser-induced fluorescence (LIF) was studied in vivo from premalignant and benign lesions in the female genital tract, in particular the cervix. The aim of the study was to investigate the possibilities to differentiate cervical intraepithelial neoplasia (CIN) from normal tissue by means of two different fluorescence modalities. Most of the patients were given a low dose (5 mg/kg bw) of (delta) -amino levulinic acid (ALA). The ALA was orally administered 2 - 4 hours prior to the investigation. During this time, the ALA is transformed to the strongly fluorescent protoporphyrin IX (PpIX) via the haem cycle. Excitation light with a wavelength of 405 nm was used to excite the PpIX fluorescence. Excess amounts of PpIX were accumulated preferentially in diseased tissue. However, the variability in the PpIX accumulation from patient to patient was large. By using excitation light at 337 nm, the endogenous fluorophores are more efficiently excited. Therefore, this excitation modality was exploited for studying spectral characteristics of the autofluorescence in different tissue types. The spectra obtained were evaluated by forming fluorescence intensity ratios. The tissue types were grouped according to the histopathological examination. A correlation with the fluorescence ratios was performed. Some problems with the classification remain, mostly due to the difficulties in obtaining histopathologic evaluation of the biopsies at the exact location of the LIF measurements.

The discovery of indicator variables for QSAR using inductive logic programming

NASA Astrophysics Data System (ADS)

King, Ross D.; Srinivasan, Ashwin

1997-11-01

A central problem in forming accurate regression equations in QSAR studies isthe selection of appropriate descriptors for the compounds under study. Wedescribe a novel procedure for using inductive logic programming (ILP) todiscover new indicator variables (attributes) for QSAR problems, and show thatthese improve the accuracy of the derived regression equations. ILP techniqueshave previously been shown to work well on drug design problems where thereis a large structural component or where clear comprehensible rules arerequired. However, ILP techniques have had the disadvantage of only being ableto make qualitative predictions (e.g. active, inactive) and not to predictreal numbers (regression). We unify ILP and linear regression techniques togive a QSAR method that has the strength of ILP at describing stericstructure, with the familiarity and power of linear regression. We evaluatedthe utility of this new QSAR technique by examining the prediction ofbiological activity with and without the addition of new structural indicatorvariables formed by ILP. In three out of five datasets examined the additionof ILP variables produced statistically better results (P < 0.01) over theoriginal description. The new ILP variables did not increase the overallcomplexity of the derived QSAR equations and added insight into possiblemechanisms of action. We conclude that ILP can aid in the process of drugdesign.

Nonpolluting automobiles

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

Hoolboom, G.J.; Szabados, B.

The advantages/disadvantages of energy storage devices, which can provide nonpolluting automobile systems are discussed. Four types of storage devices are identified: electrochemical (batteries); hydrogen; electromechanical (flywheels); and molten salt heat storage. A high-speed flywheel with a small permanent magnet motor/generator has more advantages than any of the other systems and might become a real competitor to the internal combustion engine. A flywheel/motor/generator system for automobiles now becomes practical, because of the technological advances in materials, bearings and solid state control circuits. The motor of choice is the squirrel cage induction motor, specially designed for automobile applications. The preferred controller formore » the induction motor is a forced commutated cycloconverter, which transforms a variable voltage/variable frequency source into a controlled variable-voltage/variable-frequency supply. A modulation strategy of the cycloconverter elements is selected to maintain a unity input displacement factor (power factor) under all conditions of loads voltages and frequencies. The system is similar to that of the existing automobile, if only one motor is used: master controller-controller-motor-gears (fixed)-differential-wheels. In the case of two motors, the mechanical differential is replaced by an electric one: master controller-controller-motor-gears (fixed)-wheel. A four-wheel drive vehicle is obtained when four motors with their own controllers are used. 24 refs.« less

Evaluation of the use of midazolam as a co-induction agent with ketamine for anaesthesia in sedated ponies undergoing field castration.

PubMed

Allison, A; Robinson, R; Jolliffe, C; Taylor, P M

2018-05-01

There are limited investigations comparing ketamine to a ketamine-midazolam co-induction. To compare quality and safety of general anaesthesia induced using ketamine alone with anaesthesia co-induced using ketamine and midazolam. Randomised, double blinded, placebo controlled trial. After i.v. detomidine (20 μg/kg) thirty-eight ponies undergoing field castration received either 0.06 mg/kg (0.6 mL/50 kg) midazolam (group M) or 0.6 mL/50 kg placebo (group P) with 2.2 mg/kg ketamine i.v. for anaesthetic induction. Quality of anaesthetic induction, endotracheal intubation, surgical relaxation and recovery were scored using combinations of simple descriptive and visual analogue scales. Time of sedation, induction, start of endotracheal intubation, first movement, sternal recumbency and standing were recorded, as were time, number and total quantity of additional i.v. detomidine and ketamine injections. Cardiorespiratory variables were assessed every 5 min. Adverse effects were documented. Data were tested for normality and analysed with a mixed model ANOVA, Fisher's exact test, unpaired Students' t test and Wilcoxon Rank-sum as appropriate; P<0.05 was considered significant. Group M had better scores for induction (P = 0.005), intubation (P<0.001) and surgical relaxation (P<0.001) and required fewer additional injections of detomidine and ketamine (P = 0.04). Time (minutes) from induction to first movement (P<0.001), sternal recumbency (P =< 0.001) and standing was longer (P = 0.05) in group M. Recoveries were uneventful with no difference in quality between groups (P = 0.78). Clinical study with noninvasive monitoring undertaken in field conditions. Ketamine-midazolam co-induction compared to ketamine alone improved quality of induction, ease of intubation and muscle relaxation without impacting recovery quality. © 2017 EVJ Ltd.

Apple S-RNase interacts with an actin-binding protein, MdMVG, to reduce pollen tube growth by inhibiting its actin-severing activity at the early stage of self-pollination induction.

PubMed

Yang, Qing; Meng, Dong; Gu, Zhaoyu; Li, Wei; Chen, Qiuju; Li, Yang; Yuan, Hui; Yu, Jie; Liu, Chunsheng; Li, Tianzhong

2018-04-18

In S-RNase-mediated self-incompatibility, S-RNase secreted from the style destroys the actin cytoskeleton of the self-pollen tubes, eventually halting their growth, but the mechanism of this process remains unclear. In vitro biochemical assays revealed that S-RNase does not bind or sever filamentous actin (F-actin). In apple (Malus domestica), we identified an actin-binding protein containing myosin, villin and GRAM (MdMVG), that physically interacts with S-RNase and directly binds and severs F-actin. Immunofluorescence assays and total internal reflection fluorescence microscopy indicated that S-RNase inhibits the F-actin-severing activity of MdMVG in vitro. In vivo, the addition of S-RNase to self-pollen tubes increased the fluorescence intensity of actin microfilaments and reduced the severing frequency of microfilaments and the rate of pollen tube growth in self-pollination induction in the presence of MdMVG overexpression. By generating 25 single-, double- and triple-point mutations in the amino acid motif E-E-K-E-K of MdMVG via mutagenesis and testing the resulting mutants with immunofluorescence, we identified a triple-point mutant, MdMVG (E167A/E171A/K185A) , that no longer has F-actin-severing activity or interacts with any of the four S-haplotype S-RNases, indicating that all three amino acids (E167, E171 and K185) are essential for the severing activity of MdMVG and its interaction with S-RNases. We conclude that apple S-RNase interacts with MdMVG to reduce self-pollen tube growth by inhibiting its F-actin-severing activity. © 2018 The Authors The Plant Journal © 2018 John Wiley & Sons Ltd.

Clinical applicability and prognostic significance of molecular response assessed by fluorescent-PCR of immunoglobulin genes in multiple myeloma. Results from a GEM/PETHEMA study.

PubMed

Martinez-Lopez, Joaquin; Fernández-Redondo, Elena; García-Sánz, Ramón; Montalbán, María Angeles; Martínez-Sánchez, Pilar; Pavia, Bruno; Mateos, María Victoria; Rosiñol, Laura; Martín, Marisa; Ayala, Rosa; Martínez, Rafael; Blanchard, María Jesus; Alegre, Adrian; Besalduch, Joan; Bargay, Joan; Hernandez, Miguel T; Sarasquete, María Eugenia; Sanchez-Godoy, Pedro; Fernández, Manuela; Blade, Joan; San Miguel, Jesús F; Lahuerta, Juan Jose

2013-12-01

Minimal residual disease monitoring is becoming increasingly important in multiple myeloma (MM), but multiparameter flow cytometry (MFC) and allele-specific oligonucleotide polymerase chain reaction (ASO-PCR) techniques are not routinely available. This study investigated the prognostic influence of achieving molecular response assessed by fluorescent-PCR (F-PCR) in 130 newly diagnosed MM patients from Grupo Español Multidisciplinar de Melanoma (GEM)2000/GEM05 trials (NCT00560053, NCT00443235, NCT00464217) who achieved almost very good partial response after induction therapy. As a reference, we used the results observed with simultaneous MFC. F-PCR at diagnosis was performed on DNA using three different multiplex PCRs: IGH D-J, IGK V-J and KDE rearrangements. The applicability of F-PCR was 91·5%. After induction therapy, 64 patients achieved molecular response and 66 non-molecular response; median progression-free survival (PFS) was 61 versus 36 months, respectively (P = 0·001). Median overall survival (OS) was not reached (NR) in molecular response patients (5-year survival: 75%) versus 66 months in the non-molecular response group (P = 0·03). The corresponding PFS and OS values for patients with immunophenotypic versus non-immunophenotypic response were 67 versus 42 months (P = 0·005) and NR (5-year survival: 95%) versus 69 months (P = 0·004), respectively. F-PCR analysis is a rapid, affordable, and easily performable technique that, in some circumstances, may be a valid approach for minimal residual disease investigations in MM. © 2013 John Wiley & Sons Ltd.

Green tea extract induces protective autophagy in A549 non-small lung cancer cell line.

PubMed

Izdebska, Magdalena; Klimaszewska-Wiśniewska, Anna; Hałas, Marta; Gagat, Maciej; Grzanka, Alina

2015-12-31

For many decades, polyphenols, including green tea extract catechins, have been reported to exert multiple anti-tumor activities. However, to date the mechanisms of their action have not been completely elucidated. Thus, the aim of this study was to assess the effect of green tea extract on non-small lung cancer A549 cells. A549 cells following treatment with GTE were analyzed using the inverted light and fluorescence microscope. In order to evaluate cell sensitivity and cell death, the MTT assay and Tali image-based cytometer were used, respectively. Ultrastructural alterations were assessed using a transmission electron microscope. The obtained data suggested that GTE, even at the highest dose employed (150 μM), was not toxic to A549 cells. Likewise, the treatment with GTE resulted in only a very small dose-dependent increase in the population of apoptotic cells. However, enhanced accumulation of vacuole-like structures in response to GTE was seen at the light and electron microscopic level. Furthermore, an increase in the acidic vesicular organelles and LC3-II puncta formation was observed under the fluorescence microscope, following GTE treatment. The analysis of the functional status of autophagy revealed that GTE-induced autophagy may provide self-protection against its own cytotoxicity, since we observed that the blockage of autophagy by bafilomycin A1 decreased the viability of A549 cells and potentiated necrotic cell death induction in response to GTE treatment. Collectively, our results revealed that A549 cells are insensitive to both low and high concentrations of the green tea extract, probably due to the induction of cytoprotective autophagy. These data suggest that a potential utility of GTE in lung cancer therapy may lie in its synergistic combinations with drugs or small molecules that target autophagy, rather than in monotherapy.

Chiral Thioxanthones as Modulators of P-glycoprotein: Synthesis and Enantioselectivity Studies.

PubMed

Lopes, Ana; Martins, Eva; Silva, Renata; Pinto, Madalena M M; Remião, Fernando; Sousa, Emília; Fernandes, Carla

2018-03-10

Recently, thioxanthone derivatives were found to protect cells against toxic P-glycoprotein (P-gp) substrates, acting as potent inducers/activators of this efflux pump. The study of new P-gp chiral modulators produced from thioxanthone derivatives could clarify the enantioselectivity of this ABC transporter towards this new class of modulators. The aim of this study was to evaluate the P-gp modulatory ability of four enantiomeric pairs of new synthesized chiral aminated thioxanthones (ATxs) 1 - 8 , studying the influence of the stereochemistry on P-gp induction/ activation in cultured Caco-2 cells. The data displayed that all the tested compounds (at 20 μM) significantly decreased the intracellular accumulation of a P-gp fluorescent substrate (rhodamine 123) when incubated simultaneously for 60 min, demonstrating an increased activity of the efflux, when compared to control cells. Additionally, all of them except ATx 3 (+), caused similar results when the accumulation of the P-gp fluorescent substrate was evaluated after pre-incubating cells with the test compounds for 24 h, significantly reducing the rhodamine 123 intracellular accumulation as a result of a significant increase in P-gp activity. However, ATx 2 (-) was the only derivative that, after 24 h of incubation, significantly increased P-gp expression. These results demonstrated a significantly increased P-gp activity, even without an increase in P-gp expression. Therefore, ATxs 1 - 8 were shown to behave as P-gp activators. Furthermore, no significant differences were detected in the activity of the protein when comparing the enantiomeric pairs. Nevertheless, ATx 2 (-) modulates P-gp expression differently from its enantiomer, ATx 1 (+). These results disclosed new activators and inducers of P-gp and highlight the existence of enantioselectivity in the induction mechanism.

Analysis of metal-laden water via portable X-ray fluorescence spectrometry

NASA Astrophysics Data System (ADS)

Pearson, Delaina; Weindorf, David C.; Chakraborty, Somsubhra; Li, Bin; Koch, Jaco; Van Deventer, Piet; de Wet, Jandre; Kusi, Nana Yaw

2018-06-01

A rapid method for in-situ elemental composition analysis of metal-laden water would be indispensable for studying polluted water. Current analytical lab methods to determine water quality include flame atomic absorption spectrometry (FAAS), atomic absorption spectrophotometry (AAS), electrothermal atomic absorption spectrometry (EAAS), and inductively coupled plasma (ICP) spectroscopy. However only two field methods, colorimetry and absorptiometry, exist for elemental analysis of water. Portable X-ray fluorescence (PXRF) spectrometry is an effective method for elemental analysis of soil, sediment, and other matrices. However, the accuracy of PXRF is known to be affected while scanning moisture-laden soil samples. This study sought to statistically establish PXRF's predictive ability for various elements in water at different concentrations relative to inductively coupled plasma atomic emission spectroscopy (ICP-AES). A total of 390 metal-laden water samples collected from leaching columns of mine tailings in South Africa were analyzed via PXRF and ICP-AES. The PXRF showed differential effectiveness in elemental quantification. For the collected water samples, the best relationships between ICP and PXRF elemental data were obtained for K and Cu (R2 = 0.92). However, when scanning ICP calibration solutions with elements in isolation, PXRF results indicated near perfect agreement; Ca, K, Fe, Cu and Pb produced an R2 of 0.99 while Zn and Mn produced an R2 of 1.00. The utilization of multiple PXRF (stacked) beams produced stronger correlation to ICP relative to the use of a single beam in isolation. The results of this study demonstrated the PXRF's ability to satisfactorily predict the composition of metal-laden water as reported by ICP for several elements. Additionally this study indicated the need for a "Water Mode" calibration for the PXRF and demonstrates the potential of PXRF for future study of polluted or contaminated waters.

Induction and maintenance of DNA methylation in plant promoter sequences by apple latent spherical virus-induced transcriptional gene silencing

PubMed Central

Kon, Tatsuya; Yoshikawa, Nobuyuki

2014-01-01

Apple latent spherical virus (ALSV) is an efficient virus-induced gene silencing vector in functional genomics analyses of a broad range of plant species. Here, an Agrobacterium-mediated inoculation (agroinoculation) system was developed for the ALSV vector, and virus-induced transcriptional gene silencing (VITGS) is described in plants infected with the ALSV vector. The cDNAs of ALSV RNA1 and RNA2 were inserted between the cauliflower mosaic virus 35S promoter and the NOS-T sequences in a binary vector pCAMBIA1300 to produce pCALSR1 and pCALSR2-XSB or pCALSR2-XSB/MN. When these vector constructs were agroinoculated into Nicotiana benthamiana plants with a construct expressing a viral silencing suppressor, the infection efficiency of the vectors was 100%. A recombinant ALSV vector carrying part of the 35S promoter sequence induced transcriptional gene silencing of the green fluorescent protein gene in a line of N. benthamiana plants, resulting in the disappearance of green fluorescence of infected plants. Bisulfite sequencing showed that cytosine residues at CG and CHG sites of the 35S promoter sequence were highly methylated in the silenced generation zero plants infected with the ALSV carrying the promoter sequence as well as in progeny. The ALSV-mediated VITGS state was inherited by progeny for multiple generations. In addition, induction of VITGS of an endogenous gene (chalcone synthase-A) was demonstrated in petunia plants infected with an ALSV vector carrying the native promoter sequence. These results suggest that ALSV-based vectors can be applied to study DNA methylation in plant genomes, and provide a useful tool for plant breeding via epigenetic modification. PMID:25426109

Assessing the photoaging process at sun exposed and non-exposed skin using fluorescence lifetime spectroscopy

NASA Astrophysics Data System (ADS)

Saito Nogueira, Marcelo; Kurachi, Cristina

2016-03-01

Photoaging is the skin premature aging due to exposure to ultraviolet light, which damage the collagen, elastin and can induce alterations on the skin cells DNA, and, then, it may evolve to precancerous lesions, which are widely investigated by fluorescence spectroscopy and lifetime. The fluorescence spectra and fluorescence lifetime analysis has been presented as a technique of great potential for biological tissue characterization at optical diagnostics. The main targeted fluorophores are NADH (nicotinamide adenine dinucleotide) and FAD (flavin adenine dinucleotide), which have free and bound states, each one with different average lifetimes. The average lifetimes for free and bound NADH and FAD change according to tissue metabolic alterations and may contribute to a non-invasive clinical investigation of injuries such as skin lesions. These lesions and the possible areas where they may develop can be interrogated using fluorescence lifetime spectroscopy taking into account the variability of skin phototypes and the changes related to melanin, collagen and elastin, endogenous fluorophores which have emissions that spectrally overlap to the NADH and FAD emission. The objective of this study is to assess the variation on fluorescence lifetimes of normal skin at sun exposed and non-exposed areas and associate this variation to the photoaging process.

5-ALA based photodynamic management of glioblastoma

NASA Astrophysics Data System (ADS)

Rühm, Adrian; Stepp, Herbert; Beyer, Wolfgang; Hennig, Georg; Pongratz, Thomas; Sroka, Ronald; Schnell, Oliver; Tonn, Jörg-Christian; Kreth, Friedrich-Wilhelm

2014-03-01

Objective: Improvement of the clinical outcome of glioblastoma (GBM) patients by employment of fluorescence and photosensitization on the basis of 5-aminolevulinic acid (5-ALA) induced protoporphyrin IX (PpIX). Methods: In this report the focus is laid on the use of tumor selective PpIX fluorescence for stereotactic biopsy sampling and intra-operative treatment monitoring. In addition, our current concept for treatment planning is presented. For stereotactic interstitial photodynamic therapy (iPDT), radial diffusers were implanted into the contrast enhancing tumor volume. Spectroscopic measurements of laser light transmission and fluorescence between adjacent fibers were performed prior, during and post PDT. Results: PpIX concentrations in primary glioblastoma tissue show high intra- and inter-patient variability, but are usually sufficient for an effective PDT. During individual treatment attempts with 5-ALA based GBM-iPDT, transmission and fluorescence measurements between radial diffusers gave the following results: 1. In some cases, transmission after PDT is considerably reduced compared to the value before PDT, which may be attributable to a depletion of oxygenated hemoglobin and/or diffuse bleeding. 2. PpIX fluorescence is efficiently photobleached during PDT in all cases. Conclusion: iPDT with assessment of PpIX fluorescence and photobleaching is a promising treatment option. Individualization of treatment parameters appears to bear a potential to further improve clinical outcomes.

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

Obaid, Razib; Buth, Christian; Dakovski, Georgi L.

Here, we measured the fluorescence photon yield of neon upon soft x-ray ionization (~1200 eV) from the x-ray free-electron laser at Linac Coherent Light Source, and demonstrated the usage of a grazing incidence spectrometer with a variable line spacing grating to perform x-ray fluorescence spectroscopy on a gas phase system. Our measurements also allowed us to estimate the focal size of the beam from the theoretical description developed, in terms of the rate equation approximation accounting for photoionization shake off of neutral neon and double auger decay of single core holes.

Effect of 900 MHz Electromagnetic Radiation on the Induction of ROS in Human Peripheral Blood Mononuclear Cells.

PubMed

Kazemi, E; Mortazavi, S M J; Ali-Ghanbari, A; Sharifzadeh, S; Ranjbaran, R; Mostafavi-Pour, Z; Zal, F; Haghani, M

2015-09-01

Despite numerous studies over a decade, it still remains controversial about the biological effects of RF EMF emitted by mobile phone telephony. Here we investigated the effect of 900 MHz GSM on the induction of oxidative stress and the level of intracellular reactive oxygen species (ROS) in human mononuclear cells, monocytes and lymphocytes as defence system cells. 6 ml Peripheral Blood samples were obtained from 13 healthy volunteers (21-30 year-old). Each sample was devided into 2 groups: one was exposed RF radiation emitted from a mobile phone simulator for 2 hour and the other used as control group which was not exposed to any fields. After that, mononuclear cells were isolated from peripheral blood by density gradient centrifugation in Ficoll-Paque. The intracellular ROS content in monocytes and lymphocytes was measured by the CM-H2DCFDA fluorescence probe using flowcytometry technique. Our results showed significant increase in  ROS production after exposure in population rich in monocytes. This effect was not significant in population rich in lymphocytes in comparison with non exposed cells. The results obtained in this study clearly showed the oxidative stress induction capability of RF electromagnetic field in the portion of PBMCs mostly in monocytes, like the case of exposure to micro organisms, although the advantages or disadvantages of this effect should be evaluated.

Morphology and contractile gene expression of adipose-derived mesenchymal stem cells in response to short-term cyclic uniaxial strain and TGF-β1.

PubMed

Rashidi, Neda; Tafazzoli-Shadpour, Mohammad; Haghighipour, Nooshin; Khani, Mohammad-Mehdi

2018-06-27

Previous studies have shown smooth muscle induction in adipose-derived mesenchymal stem cells (ASCs) caused by long-term cyclic stretch. Here we examined the capability of the short-term straining with time steps of 4, 8, 16 and 24 h alone or combined with TGF-β1 on smooth muscle induction of rabbit ASCs. Alterations in cell morphology were quantified through the cell shape index and orientation angle, and expression levels of α-SMA, SM22-α, h-caldesmon and calponin3 markers were examined using the real-time polymerase chain reaction (PCR) method. Moreover, F-actin cytoskeleton organization was observed by fluorescence staining. Mechanical strain either alone or combined with growth factor treatment caused significant up-regulation of both early and intermediate smooth muscle cells (SMCs) specific markers during the initial hours of stimulation peaking in 8 to 16 h. Furthermore, gradual alignment of cells perpendicular to the strain direction during loading time, and cell elongation resembling contractile SMC phenotype, together with alignment and reorganization of F-actin fibers were observed. Considering previously reported protein up-regulation in following days of straining, the effects of short-term cyclic stretch on smooth muscle induction of ASCs were revealed which can be helpful in achieving functional contractile SMCs through synergistic mechano-chemical regulation of ASCs as an appealing cell source for vascular tissue engineering.

Effect of 900 MHz Electromagnetic Radiation on the Induction of ROS in Human Peripheral Blood Mononuclear Cells

PubMed Central

Kazemi, E.; Mortazavi, S. M. J.; Ali-Ghanbari, A.; Sharifzadeh, S.; Ranjbaran, R.; Mostafavi-pour, Z.; Zal, F.; Haghani, M.

2015-01-01

Background Despite numerous studies over a decade, it still remains controversial about the biological effects of RF EMF emitted by mobile phone telephony. Objective Here we investigated the effect of 900 MHz GSM on the induction of oxidative stress and the level of intracellular reactive oxygen species (ROS) in human mononuclear cells, monocytes and lymphocytes as defence system cells. Method 6 ml Peripheral Blood samples were obtained from 13 healthy volunteers (21-30 year-old). Each sample was devided into 2 groups: one was exposed RF radiation emitted from a mobile phone simulator for 2 hour and the other used as control group which was not exposed to any fields. After that, mononuclear cells were isolated from peripheral blood by density gradient centrifugation in Ficoll-Paque. The intracellular ROS content in monocytes and lymphocytes was measured by the CM-H2DCFDA fluorescence probe using flowcytometry technique. Results Our results showed significant increase in  ROS production after exposure in population rich in monocytes. This effect was not significant in population rich in lymphocytes in comparison with non exposed cells. Conclusion The results obtained in this study clearly showed the oxidative stress induction capability of RF electromagnetic field in the portion of PBMCs mostly in monocytes, like the case of exposure to micro organisms, although the advantages or disadvantages of this effect should be evaluated. PMID:26396966

Trehalose, sucrose and raffinose are novel activators of autophagy in human keratinocytes through an mTOR-independent pathway

PubMed Central

Chen, Xu; Li, Min; Li, Li; Xu, Song; Huang, Dan; Ju, Mei; Huang, Ju; Chen, Kun; Gu, Heng

2016-01-01

Trehalose is a natural disaccharide that is found in a diverse range of organisms but not in mammals. Autophagy is a process which mediates the sequestration, lysosomal delivery and degradation of proteins and organelles. Studies have shown that trehalose exerts beneficial effects through inducing autophagy in mammalian cells. However, whether trehalose or other saccharides can activate autophagy in keratinocytes is unknown. Here, we found that trehalose treatment increased the LC3-I to LC3-II conversion, acridine orange-stained vacuoles and GFP-LC3B (LC3B protein tagged with green fluorescent protein) puncta in the HaCaT human keratinocyte cell line, indicating autophagy induction. Trehalose-induced autophagy was also observed in primary keratinocytes and the A431 epidermal cancer cell line. mTOR signalling was not affected by trehalose treatment, suggesting that trehalose induced autophagy through an mTOR-independent pathway. mTOR-independent autophagy induction was also observed in HaCaT and HeLa cells treated with sucrose or raffinose but not in glucose, maltose or sorbitol treated HaCaT cells, indicating that autophagy induction was not a general property of saccharides. Finally, although trehalose treatment had an inhibitory effect on cell proliferation, it had a cytoprotective effect on cells exposed to UVB radiation. Our study provides new insight into the saccharide-mediated regulation of autophagy in keratinocytes. PMID:27328819

A Member of the p38 Mitogen-Activated Protein Kinase Family Is Responsible for Transcriptional Induction of Dopa decarboxylase in the Epidermis of Drosophila melanogaster during the Innate Immune Response▿ †

PubMed Central

Davis, Monica M.; Primrose, David A.; Hodgetts, Ross B.

2008-01-01

Drosophila innate immunity is controlled primarily by the activation of IMD (immune deficiency) or Toll signaling leading to the production of antimicrobial peptides (AMPs). IMD signaling also activates the JUN N-terminal kinase (JNK) cascade, which is responsible for immune induction of non-antimicrobial peptide immune gene transcription though the transcription factor AP-1. Transcription of the Dopa decarboxylase (Ddc) gene is induced in response to gram-negative and gram-positive septic injury, but not aseptic wounding. Transcription is induced throughout the epidermis and not specifically at the site of infection. Ddc transcripts are detectible within 2 h and remain high for several hours following infection with either gram-negative or gram-positive bacteria. Using Ddc-green fluorescent protein (GFP) reporter gene constructs, we show that a conserved consensus AP-1 binding site upstream of the Ddc transcription start site is required for induction. However, neither the Toll, IMD, nor JNK pathway is involved. Rather, Ddc transcription depends on a previously uncharacterized member of the p38 mitogen-activated protein kinase family, p38c. We propose that the involvement of DDC in a new pathway involved in Drosophila immunity increases the levels of dopamine, which is metabolized to produce reactive quinones that exert an antimicrobial effect on invading bacteria. PMID:18519585

Re-evaluation and extension of the scope of elements in US Geological Survey Standard Reference Water Samples

USGS Publications Warehouse

Peart, D.B.; Antweiler, Ronald C.; Taylor, Howard E.; Roth, D.A.; Brinton, T.I.

1998-01-01

More than 100 US Geological Survey (USGS) Standard Reference Water Samples (SRWSs) were analyzed for numerous trace constituents, including Al, As, B, Ba, Be, Bi, Br, Cd, Cr, Co, Cu, I, Fe, Pb, Li, Mn, Mo, Ni, Rb, Sb, Se, Sr, Te, Tl, U, V, Zn and major elements (Ca, Mg, Na, SiO2, SO4, Cl) by inductively coupled plasma mass spectrometry and inductively coupled plasma atomic emission spectrometry. In addition, 15 USGS SRWSs and National Institute of Standards and Technology (NIST) standard reference material (SRM) 1641b were analyzed for mercury using cold vapor atomic fluorescence spectrometry. Also USGS SRWS Hg-7 was analyzed using isotope dilution-inductively coupled plasma mass spectrometry. The results were compared with the reported certified values of the following standard reference materials: NIST SRM 1643a, 1643b, 1643c and 1643d and National Research Council of Canada Riverine Water Reference Materials for Trace Metals SLRS-1, SLRS-2 and SLRS-3. New concentration values for trace and major elements in the SRWSs, traceable to the certified standards, are reported. Additional concentration values are reported for elements that were neither previously published for the SRWSs nor traceable to the certified reference materials. Robust statistical procedures were used that were insensitive to outliers. These data can be used for quality assurance/quality control purposes in analytical laboratories.

Fuel saver based on electromagnetic induction for automotive engine

NASA Astrophysics Data System (ADS)

Siregar, Houtman P.; Sibarani, Maradu

2007-12-01

In the considered research is designed and analyzed the performance of the fuel saver which is based on electromagnetic induction for automotive diesel engine. The fuel saver which is based on permanent magnet has sold in market and its performance has tested. In comparison to the former fuel saver, in the proposed work is produced fuel saver which is based on electromagnetic induction. The considered research is the continuation of my former work. Performance of the produced fuel saver which is installed in the fuel line of internal combustion engine rig is compared to the performance of the standard internal combustion engine rig Speed of the engine, wire diameter of coil, and number of coil which is coiled in the winding of the the fuel saver are chosen as the testing variables. The considered research has succeeded to design the fuel saver which is based on electromagnetic induction for saving the automotive fuel consumption. Results of the research show that the addition of the fuel saver which is based on electromagnetic induction to the flow of the diesel fuel can significantly save the automative fuel consumption. In addition the designed fuel saver can reduce the opacity of the emission gas.

Confirming the Multidimensionality of Psychologically Controlling Parenting among Chinese-American Mothers: Love Withdrawal, Guilt Induction, and Shaming.

PubMed

Cheah, Charissa; Yu, Jing; Hart, Craig; Sun, Shuyan; Olsen, Joseph

2015-05-01

Despite the theoretical conceptualization of parental psychological control as a multidimensional construct, the majority of previous studies have examined psychological control as a unidimensional scale. Moreover, the conceptualization of shaming and its associations with love withdrawal and guilt induction are unclear. The current study aimed to fill these gaps by evaluating the latent factor structure underlying 18 items from Olsen et al. (2002) that were conceptually relevant to love withdrawal, guilt induction, and shaming practices in a sample of 169 mothers of Chinese-American preschoolers. A multidimensional three-factor model and bi-factor model were specified based on our formulated operational definitions for the three dimensions of psychological control. Both models were found to be superior to the unidimensional model. In addition, results from the bi-factor model and an additional second-order factor model indicated that psychological control is essentially empirically isomorphic with guilt induction. Although love withdrawal and shaming factors were also fairly strong indicators of psychological control, each exhibited important additional unique variability and mutual distinctiveness. Implications for the conceptualization of love withdrawal, guilt induction, and shaming as well as directions for future studies are discussed.

Confirming the Multidimensionality of Psychologically Controlling Parenting among Chinese-American Mothers: Love Withdrawal, Guilt Induction, and Shaming

PubMed Central

Cheah, Charissa; Yu, Jing; Hart, Craig; Sun, Shuyan; Olsen, Joseph

2014-01-01

Despite the theoretical conceptualization of parental psychological control as a multidimensional construct, the majority of previous studies have examined psychological control as a unidimensional scale. Moreover, the conceptualization of shaming and its associations with love withdrawal and guilt induction are unclear. The current study aimed to fill these gaps by evaluating the latent factor structure underlying 18 items from Olsen et al. (2002) that were conceptually relevant to love withdrawal, guilt induction, and shaming practices in a sample of 169 mothers of Chinese-American preschoolers. A multidimensional three-factor model and bi-factor model were specified based on our formulated operational definitions for the three dimensions of psychological control. Both models were found to be superior to the unidimensional model. In addition, results from the bi-factor model and an additional second-order factor model indicated that psychological control is essentially empirically isomorphic with guilt induction. Although love withdrawal and shaming factors were also fairly strong indicators of psychological control, each exhibited important additional unique variability and mutual distinctiveness. Implications for the conceptualization of love withdrawal, guilt induction, and shaming as well as directions for future studies are discussed. PMID:26052168

Hybrid Imaging Labels: Providing the Link Between Mass Spectrometry-Based Molecular Pathology and Theranostics

PubMed Central

Buckle, Tessa; van der Wal, Steffen; van Malderen, Stijn J.M.; Müller, Larissa; Kuil, Joeri; van Unen, Vincent; Peters, Ruud J.B.; van Bemmel, Margaretha E.M.; McDonnell, Liam A.; Velders, Aldrik H.; Koning, Frits; Vanhaeke, Frank; van Leeuwen, Fijs W. B.

2017-01-01

Background: Development of theranostic concepts that include inductively coupled plasma mass spectrometry (ICP-MS) and laser ablation ICP-MS (LA-ICP-MS) imaging can be hindered by the lack of a direct comparison to more standardly used methods for in vitro and in vivo evaluation; e.g. fluorescence or nuclear medicine. In this study a bimodal (or rather, hybrid) tracer that contains both a fluorescent dye and a chelate was used to evaluate the existence of a direct link between mass spectrometry (MS) and in vitro and in vivo molecular imaging findings using fluorescence and radioisotopes. At the same time, the hybrid label was used to determine whether the use of a single isotope label would allow for MS-based diagnostics. Methods: A hybrid label that contained both a DTPA chelate (that was coordinated with either 165Ho or 111In) and a Cy5 fluorescent dye was coupled to the chemokine receptor 4 (CXCR4) targeting peptide Ac-TZ14011 (hybrid-Cy5-Ac-TZ4011). This receptor targeting tracer was used to 1) validate the efficacy of (165Ho-based) mass-cytometry in determining the receptor affinity via comparison with fluorescence-based flow cytometry (Cy5), 2) evaluate the microscopic binding pattern of the tracer in tumor cells using both fluorescence confocal imaging (Cy5) and LA-ICP-MS-imaging (165Ho), 3) compare in vivo biodistribution patterns obtained with ICP-MS (165Ho) and radiodetection (111In) after intravenous administration of hybrid-Cy5-Ac-TZ4011 in tumor-bearing mice. Finally, LA-ICP-MS-imaging (165Ho) was linked to fluorescence-based analysis of excised tissue samples (Cy5). Results: Analysis with both mass-cytometry and flow cytometry revealed a similar receptor affinity, respectively 352 ± 141 nM and 245 ± 65 nM (p = 0.08), but with a much lower detection sensitivity for the first modality. In vitro LA-ICP-MS imaging (165Ho) enabled clear discrimination between CXCR4 positive and negative cells, but fluorescence microscopy was required to determine the intracellular distribution. In vivo biodistribution patterns obtained with ICP-MS (165Ho) and radiodetection (111In) of the hybrid peptide were shown to be similar. Assessment of tracer distribution in excised tissues revealed the location of tracer uptake with both LA-ICP-MS-imaging and fluorescence imaging. Conclusion: Lanthanide-isotope chelation expands the scope of fluorescent/radioactive hybrid tracers to include MS-based analytical tools such as mass-cytometry, ICP-MS and LA-ICP-MS imaging in molecular pathology. In contradiction to common expectations, MS detection using a single chelate imaging agent was shown to be feasible, enabling a direct link between nuclear medicine-based imaging and theranostic methods. PMID:28255355

Spatial variability of concentrations of chlorophyll a, dissolved organic matter and suspended particles in the surface layer of the Kara Sea in September 2011 from lidar data

NASA Astrophysics Data System (ADS)

Pelevin, V. V.; Zavjalov, P. O.; Belyaev, N. A.; Konovalov, B. V.; Kravchishina, M. D.; Mosharov, S. A.

2017-01-01

The article presents results of underway remote laser sensing of the surface water layer in continuous automatic mode using the UFL-9 fluorescent lidar onboard the R/V Akademik Mstislav Keldysh during cruise 59 in the Kara Sea in 2011. The description of the lidar, the approach to interpreting seawater fluorescence data, and certain methodical aspects of instrument calibration and measurement are presented. Calibration of the lidar is based on laboratory analysis of water samples taken from the sea surface during the cruise. Spatial distribution of chlorophyll a, total organic carbon and suspended matter concentrations in the upper quasi-homogeneous layer are mapped and the characteristic scales of the variability are estimated. Some dependencies between the patchiness of the upper water layer and the atmospheric forcing and freshwater runoff are shown.

Rapid determination of vitellogenin in fish plasma by anion exchange high performance liquid chromatography using postcolumn fluorescence derivatization with o-phthalaldehyde.

PubMed

Wu, Cuiqin; Yuan, Dongxing; Liu, Baomin

2006-12-01

An analytical method involving anion exchange high performance liquid chromatographic determination of vitellogenin (Vtg) in fish plasma after postcolumn fluorescence derivatization with o-phthalaldehyde (OPA) was developed. The retention time of Vtg was about 11 min. The reagent variables for derivatization were optimized. The fluorophore was excited at 335 nm and detected at 435 nm. A calibration curve was established ranging from 0.13 to 11.28 microg. The determination limit of Vtg was found to be as low as 0.13 microg. The spiked recovery was 93.6% and interassay variability was less than 4%. The method developed was used to determine Vtg in fish plasma obtained from red sea bream (Pagrosomus major), black porgy (Sparus macrocephalus) and skew band grunt (Hapalogenys nitens), without complicated sample pretreatment. The results confirmed that the method showed advantages of being simple, rapid, reproducible and sensitive.

An attempt to detect lameness in galloping horses by use of body-mounted inertial sensors.

PubMed

Lopes, Marco A F; Dearo, Antonio C O; Lee, Allen; Reed, Shannon K; Kramer, Joanne; Pai, P Frank; Yonezawa, Yoshiharu; Maki, Hiromitchi; Morgan, Terry L; Wilson, David A; Keegan, Kevin G

2016-10-01

OBJECTIVE To evaluate head, pelvic, and limb movement to detect lameness in galloping horses. ANIMALS 12 Thoroughbreds. PROCEDURES Movement data were collected with inertial sensors mounted on the head, pelvis, and limbs of horses trotting and galloping in a straight line before and after induction of forelimb and hind limb lameness by use of sole pressure. Successful induction of lameness was determined by measurement of asymmetric vertical head and pelvic movement during trotting. Differences in gallop strides before and after induction of lameness were evaluated with paired-sample statistical analysis and neural network training and testing. Variables included maximum, minimum, range, and time indices of vertical head and pelvic acceleration, head rotation in the sagittal plane, pelvic rotation in the frontal plane, limb contact intervals, stride durations, and limb lead preference. Difference between median standardized gallop strides for each limb lead before and after induction of lameness was calculated as the sum of squared differences at each time index and assessed with a 2-way ANOVA. RESULTS Head and pelvic acceleration and rotation, limb timing, stride duration measurements, and limb lead preference during galloping were not significantly different before and after induction of lameness in the forelimb or hind limb. Differences between limb leads before induction of lameness were similar to or greater than differences within limb leads before and after lameness induction. CONCLUSIONS AND CLINICAL RELEVANCE Galloping horses maintained asymmetry of head, pelvic, and limb motion between limb leads that was unrelated to lameness.

Analysis of quetiapine in human plasma using fluorescence spectroscopy

NASA Astrophysics Data System (ADS)

Mostafa, Islam M.; Omar, Mahmoud A.; Nagy, Dalia M.; Derayea, Sayed M.

2018-05-01

A simple and sensitive spectrofluorimetric method has been development for the assurance of quetiapine fumarate (QTF). The proposed method was utilized for measuring the fluorescence intensity of the yellow fluorescent product at 510 nm (λex 470 nm). The fluorescent product has resulted from the nucleophilic substitution reaction of QTF with 4-chloro-7-nitrobenzofurazane (NBD-Cl) in Mcllvaine buffer (pH 7.0). The diverse variables influencing the development of the reaction product were deliberately changed and optimized. The linear concentration range of the proposed method was of 0.2-2.0 μg ml-1.The limits of detection and quantitation were 0.05 and 0.17 μg ml-1, respectively. The proposed method was applied for the assurance of QTF in its tablets without interference from basic excipients. In addition, the proposed method was used for in vitro analysis of the QTF in spiked human plasma, the percent mean recovery was (n = 3) 98.82 ± 1.484%.

2-dimensional ion velocity distributions measured by laser-induced fluorescence above a radio-frequency biased silicon wafer

NASA Astrophysics Data System (ADS)

Moore, Nathaniel B.; Gekelman, Walter; Pribyl, Patrick; Zhang, Yiting; Kushner, Mark J.

2013-08-01

The dynamics of ions traversing sheaths in low temperature plasmas are important to the formation of the ion energy distribution incident onto surfaces during microelectronics fabrication. Ion dynamics have been measured using laser-induced fluorescence (LIF) in the sheath above a 30 cm diameter, 2.2 MHz-biased silicon wafer in a commercial inductively coupled plasma processing reactor. The velocity distribution of argon ions was measured at thousands of positions above and radially along the surface of the wafer by utilizing a planar laser sheet from a pulsed, tunable dye laser. Velocities were measured both parallel and perpendicular to the wafer over an energy range of 0.4-600 eV. The resulting fluorescence was recorded using a fast CCD camera, which provided resolution of 0.4 mm in space and 30 ns in time. Data were taken at eight different phases during the 2.2 MHz cycle. The ion velocity distributions (IVDs) in the sheath were found to be spatially non-uniform near the edge of the wafer and phase-dependent as a function of height. Several cm above the wafer the IVD is Maxwellian and independent of phase. Experimental results were compared with simulations. The experimental time-averaged ion energy distribution function as a function of height compare favorably with results from the computer model.

Development of a novel method for quantification of autophagic protein degradation by AHA labeling.

PubMed

Zhang, Jianbin; Wang, Jigang; Ng, Shukie; Lin, Qingsong; Shen, Han-Ming

2014-05-01

Autophagy is a catabolic process during which cellular components including protein aggregates and organelles are degraded via a lysosome-dependent process to sustain metabolic homeostasis during nutrient or energy deprivation. Measuring the rate of proteolysis of long-lived proteins is a classical assay for measurement of autophagic flux. However, traditional methods, such as a radioisotope labeling assay, are technically tedious and have low sensitivity. Here, we report a novel method for quantification of long-lived protein degradation based on L-azidohomoalanine (AHA) labeling in mouse embryonic fibroblasts (MEFs) and in human cancer cells. AHA is a surrogate for L-methionine, containing a bio-orthogonalazide moiety. When added to cultured cells, AHA is incorporated into proteins during active protein synthesis. After a click reaction between an azide and an alkyne, the azide-containing proteins can be detected with an alkyne-tagged fluorescent dye, coupled with flow cytometry. Induction of autophagy by starvation or mechanistic target of rapamycin (MTOR) inhibitors was able to induce a significant reduction of the fluorescence intensity, consistent with other autophagic markers. Coincidently, inhibition of autophagy by pharmacological agents or by Atg gene deletion abolished the reduction of the fluorescence intensity. Compared with the classical radioisotope pulse-labeling method, we think that our method is sensitive, quantitative, nonradioactive, and easy to perform, and can be applied to both human and animal cell culture systems.

Development of a novel method for quantification of autophagic protein degradation by AHA labeling

PubMed Central

Zhang, Jianbin; Wang, Jigang; Ng, Shukie; Lin, Qingsong; Shen, Han-Ming

2014-01-01

Autophagy is a catabolic process during which cellular components including protein aggregates and organelles are degraded via a lysosome-dependent process to sustain metabolic homeostasis during nutrient or energy deprivation. Measuring the rate of proteolysis of long-lived proteins is a classical assay for measurement of autophagic flux. However, traditional methods, such as a radioisotope labeling assay, are technically tedious and have low sensitivity. Here, we report a novel method for quantification of long-lived protein degradation based on L-azidohomoalanine (AHA) labeling in mouse embryonic fibroblasts (MEFs) and in human cancer cells. AHA is a surrogate for L-methionine, containing a bio-orthogonalazide moiety. When added to cultured cells, AHA is incorporated into proteins during active protein synthesis. After a click reaction between an azide and an alkyne, the azide-containing proteins can be detected with an alkyne-tagged fluorescent dye, coupled with flow cytometry. Induction of autophagy by starvation or mechanistic target of rapamycin (MTOR) inhibitors was able to induce a significant reduction of the fluorescence intensity, consistent with other autophagic markers. Coincidently, inhibition of autophagy by pharmacological agents or by Atg gene deletion abolished the reduction of the fluorescence intensity. Compared with the classical radioisotope pulse-labeling method, we think that our method is sensitive, quantitative, nonradioactive, and easy to perform, and can be applied to both human and animal cell culture systems. PMID:24675368

An automated microreactor for semi-continuous biosensor measurements.

PubMed

Buffi, Nina; Beggah, Siham; Truffer, Frederic; Geiser, Martial; van Lintel, Harald; Renaud, Philippe; van der Meer, Jan Roelof

2016-04-21

Living bacteria or yeast cells are frequently used as bioreporters for the detection of specific chemical analytes or conditions of sample toxicity. In particular, bacteria or yeast equipped with synthetic gene circuitry that allows the production of a reliable non-cognate signal (e.g., fluorescent protein or bioluminescence) in response to a defined target make robust and flexible analytical platforms. We report here how bacterial cells expressing a fluorescence reporter ("bactosensors"), which are mostly used for batch sample analysis, can be deployed for automated semi-continuous target analysis in a single concise biochip. Escherichia coli-based bactosensor cells were continuously grown in a 13 or 50 nanoliter-volume reactor on a two-layered polydimethylsiloxane-on-glass microfluidic chip. Physiologically active cells were directed from the nl-reactor to a dedicated sample exposure area, where they were concentrated and reacted in 40 minutes with the target chemical by localized emission of the fluorescent reporter signal. We demonstrate the functioning of the bactosensor-chip by the automated detection of 50 μgarsenite-As l(-1) in water on consecutive days and after a one-week constant operation. Best induction of the bactosensors of 6-9-fold to 50 μg l(-1) was found at an apparent dilution rate of 0.12 h(-1) in the 50 nl microreactor. The bactosensor chip principle could be widely applicable to construct automated monitoring devices for a variety of targets in different environments.

Exposure of zebra mussels to extracorporeal shock waves demonstrates formation of new mineralized tissue inside and outside the focus zone.

PubMed

Sternecker, Katharina; Geist, Juergen; Beggel, Sebastian; Dietz-Laursonn, Kristin; de la Fuente, Matias; Frank, Hans-Georg; Furia, John P; Milz, Stefan; Schmitz, Christoph

2018-04-03

The success rate of extracorporeal shock wave therapy (ESWT) for fracture nonunions in human medicine (i.e., radiographic union at six months after ESWT) is only approximately 75%. Detailed knowledge regarding the underlying mechanisms that induce bio-calcification after ESWT is limited. We analyzed the biological response within mineralized tissue of a new invertebrate model organism, the zebra mussel Dreissena polymorpha , after exposure with extracorporeal shock waves (ESWs). Mussels were exposed to ESWs with positive energy density of 0.4 mJ/mm 2 (A) or were sham exposed (B). Detection of newly calcified tissue was performed by exposing the mussels to fluorescent markers. Two weeks later, the A-mussels showed a higher mean fluorescence signal intensity within the shell zone than the B-mussels (p<0.05). Acoustic measurements revealed that the increased mean fluorescence signal intensity within the shell of the A-mussels was independent of the size and position of the focal point of the ESWs. These data demonstrate that induction of bio-calcification after ESWT may not be restricted to the region of direct energy transfer of ESWs into calcified tissue. The results of the present study are of relevance for better understanding of the molecular and cellular mechanisms that induce formation of new mineralized tissue after ESWT. © 2018. Published by The Company of Biologists Ltd.

Lipid droplets formation in human endothelial cells in response to polyunsaturated fatty acids and 1-methyl-nicotinamide (MNA); confocal Raman imaging and fluorescence microscopy studies.

PubMed

Majzner, Katarzyna; Chlopicki, Stefan; Baranska, Malgorzata

2016-04-01

In this work the formation of lipid droplets (LDs) in human endothelial cells culture in response to the uptake of polyunsaturated fatty acids (PUFAs) was studied. Additionally, an effect of 1-methylnicotinamide (MNA) on the process of LDs formation was investigated. LDs have been previously described structurally and to some degree biochemically, however neither the precise function of LDs nor the factors responsible for LD induction have been clarified. Lipid droplets, sometimes referred in the literature as lipid bodies are organelles known to regulate neutrophil, eosinophil, or tumor cell functions but their presence and function in the endothelium is largely unexplored. 3D linear Raman spectroscopy was used to study LDs formation in vitro in a single endothelial cell. The method provides information about distribution and size of LDs as well as their composition. The incubation of endothelial cells with various PUFAs resulted in formation of LDs. As a complementary method for LDs identification a fluorescence microscopy was applied. Fluorescence measurements confirmed the Raman results suggesting endothelial cells uptake of PUFAs and subsequent LDs formation in the cytoplasm of the endothelium. Furthermore, MNA seem to potentiate intracellular uptake of PUFAs to the endothelium that may bear physiological and pharmacological significance. Confocal Raman imaging of HAoEC cell with LDs. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

In vivo visualization of endogenous miR-21 using hyaluronic acid-coated graphene oxide for targeted cancer therapy.

PubMed

Hwang, Do Won; Kim, Han Young; Li, Fangyuan; Park, Ji Yong; Kim, Dohyun; Park, Jae Hyung; Han, Hwa Seung; Byun, Jung Woo; Lee, Yun-Sang; Jeong, Jae Min; Char, Kookheon; Lee, Dong Soo

2017-03-01

Oncogene-targeted nucleic acid therapy has been spotlighted as a new paradigm for cancer therapeutics. However, in vivo delivery issues and uncertainty of therapeutic antisense drug reactions remain critical hurdles for a successful targeted cancer therapy. In this study, we developed a fluorescence-switchable theranostic nanoplatform using hyaluronic acid (HA)-conjugated graphene oxide (GO), which is capable of both sensing oncogenic miR-21 and inhibiting its tumorigenicity simultaneously. Cy3-labeled antisense miR-21 peptide nucleic acid (PNA) probes loaded onto HA-GO (HGP21) specifically targeted CD44-positive MBA-MB231 cells and showed fluorescence recovery by interacting with endogenous miR-21 in the cytoplasm of the MBA-MB231 cells. Knockdown of endogenous miR-21 by HGP21 led to decreased proliferation and reduced migration of cancer cells, as well as the induction of apoptosis, with enhanced PTEN levels. Interestingly, in vivo fluorescence signals markedly recovered 3 h after the intravenous delivery of HGP21 and displayed signals more than 5-fold higher than those observed in the HGPscr-treated group of tumor-bearing mice. These findings demonstrate the possibility of using the HGP nanoplatform as a cancer theranostic tool in miRNA-targeted therapy. Copyright © 2017 Elsevier Ltd. All rights reserved.

Development of a keratinocyte-based screening model for antipsoriatic drugs using green fluorescent protein under the control of an endogenous promoter.

PubMed

Pol, Arno; van Ruissen, Fred; Schalkwijk, Joost

2002-08-01

Inflamed epidermis (psoriasis, wound healing, ultraviolet-irradiated skin) harbors keratinocytes that are hyperproliferative and display an abnormal differentiation program. A distinct feature of this so-called regenerative maturation pathway is the expression of proteins such as the cytokeratins CK6, CK16, and CK17 and the antiinflammatory protein SKALP/elafin. These proteins are absent in normal skin but highly induced in lesional psoriatic skin. Expression of these genes can be used as a surrogate marker for psoriasis in drug-screening procedures of large compound libraries. The aim of this study was to develop a keratinocyte cell line that contained a reporter gene under the control of a psoriasis-associated endogenous promoter and demonstrate its use in an assay suitable for screening. We generated a stably transfected keratinocyte cell line that expresses enhanced green fluorescent protein (EGFP), under the control of a 0.8-kb fragment derived from the promoter of the SKALP/elafin gene, which confers high levels of tissue-specific expression at the mRNA level. Induction of the SKALP promoter by tumor necrosis factor-alpha resulted in increased expression levels of the secreted SKALP-EGFP fusion protein as assessed by direct readout of fluorescence and fluorescence polarization in 96-well cell culture plates. The fold stimulation of the reporter gene was comparable to that of the endogenous SKALP gene as assessed by enzyme-linked immunosorbent assay. Although the dynamic range of the screening system is limited, the small standard deviation yields a Z factor of 0.49. This indicates that the assay is suitable as a high-throughput screen, and provides proof of the concept that a secreted EGFP fusion protein under the control of a physiologically relevant endogenous promoter can be used as a fluorescence-based high-throughput screen for differentiation-modifying or antiinflammatory compounds that act via the keratinocyte.