Flow injection on-line oxidizing fluorometry coupled to dialysis sampling for the study of carbamazepine-protein binding

Energy Technology Data Exchange (ETDEWEB)

Zhang Zhiqi [School of Chemistry and Materials Science, Shaanxi Normal University, Xi' an 710062 (China)]. E-mail: zqzhang@snnu.edu.cn; Liang Guoxi [School of Chemistry and Materials Science, Shaanxi Normal University, Xi' an 710062 (China)

2005-04-22

The mechanism of binding of carbamazepine (CBZ) with bovine serum albumin (BSA) has been investigated in vitro based on a new flow injection fluorometry coupled to the technique of dialysis sampling. The CBZ and BSA were mixed in different molar ratios in 0.050 mol L{sup -1} phosphate buffer (containing 0.9% NaCl), pH 7.4, and incubated at 37 {+-} 0.5 deg. C in a water bath. The dialysis sampler was utilized to sample free CBZ from mixed solution with a relative dialytic efficiency of 7.6%. Then the CBZ in dialysis solution was injected into carrier and on-line oxidized by lead dioxide solid-phase reactor into fluorescent product with a maximum excitation wavelength of 355 nm and a maximum emission wavelength of 478 nm. The fluorescence intensity measured was linear proportional with the concentration of free CBZ in mixed solution over the range of 1 x 10{sup -5} to 2 x 10{sup -4} mol L{sup -1} with the detection limit of 6 x 10{sup -6} mol L{sup -1}. According to the fluorescence measurement results from mixed solution, the association constant (K) estimated for CBZ-BSA binding and the number of the binding site (n) with Scatchard analysis were 1.08 x 10{sup 4} L mol{sup -1} and 0.94, respectively. Stern-Volmer plots indicated the presence of dynamic component in the quenching mechanism. The acting force was suggested to be mainly hydrophobic and the distance between the acceptor and donor was 3.12 nm. The estimated binding parameters agreed well with literature values.

Comparative studies of HPLC-fluorometry and LC/MS method for the determination of N-acetylneuraminic acid as a marker of deteriorated ophthalmic solutions.

Science.gov (United States)

Iwatsuka, Kinya; Yasueda, Shin-ichi; Bando, Eiji; Fujii, Hiroyuki; Terada, Takashi; Okubo, Hiroya; Iwamoto, Hiroki; Kinoshita, Mitsuhiro; Kakehi, Kazuaki

2011-10-01

Methods for determining the deterioration of ophthalmic solutions using both high-performance liquid chromatography (HPLC) with fluorescence detection and liquid chromatography coupled with selected ion monitoring mass spectrometry (LC/MS) are described. The methods are based on the determination of N-acetylneuraminic acid (NeuAc) released by the hydrolysis of foreign bodies that contaminate eye drops during use. The released NeuAc was either labeled with 1,2-diamino-4,5-methylenedioxybenzene (DMB) for fluorometric detection or detected without derivatization by mass spectrometry. The calibration curves for NeuAc showed good linearity between 1.2 ng/mL and 39 ng/mL for fluorometric HPLC and 5.0 ng/mL and 100 ng/mL for LC/MS, respectively. Detection limits for fluorometric HPLC and LC/MS were 0.20 ng/mL and 0.88 ng/mL, respectively. The NeuAc content of some model glycoproteins determined by LC/MS method were 62-78% of those determined by fluorometry. The differences are attributed to matrix effects but the LC/MS method afforded sufficiently high sensitivity that NeuAc in the foreign bodies could be determined in eight of nine test samples. Copyright © 2011 Elsevier B.V. All rights reserved.

Negative response of photosynthesis to natural and projected high seawater temperatures estimated by pulse amplitude modulation fluorometry in a temperate coral.

Science.gov (United States)

Caroselli, Erik; Falini, Giuseppe; Goffredo, Stefano; Dubinsky, Zvy; Levy, Oren

2015-01-01

Balanophyllia europaea is a shallow water solitary zooxanthellate coral, endemic to the Mediterranean Sea. Extensive field studies across a latitudinal temperature gradient highlight detrimental effects of rising temperatures on its growth, demography, and skeletal characteristics, suggesting that depression of photosynthesis at high temperatures might cause these negative effects. Here we test this hypothesis by analyzing, by means of pulse amplitude modulation fluorometry, the photosynthetic efficiency of B. europaea specimens exposed in aquaria to the annual range of temperatures experienced in the field (13, 18, and 28°C), and two extreme temperatures expected for 2100 as a consequence of global warming (29 and 32°C). The indicators of photosynthetic performance analyzed (maximum and effective quantum yield) showed that maximum efficiency was reached at 20.0-21.6°C, slightly higher than the annual mean temperature in the field (18°C). Photosynthetic efficiency decreased from 20.0 to 13°C and even more strongly from 21.6 to 32°C. An unusual form of bleaching was observed, with a maximum zooxanthellae density at 18°C that strongly decreased from 18 to 32°C. Chlorophyll a concentration per zooxanthellae cell showed an opposite trend as it was minimal at 18°C and increased from 18 to 32°C. Since the areal chlorophyll concentration is the product of the zooxanthellae density and its cellular content, these trends resulted in a homogeneous chlorophyll concentration per coral surface across temperature treatments. This confirms that B. europaea photosynthesis is progressively depressed at temperatures >21.6°C, supporting previous hypotheses raised by the studies on growth and demography of this species. This study also confirms the threats posed to this species by the ongoing seawater warming.

Negative response of photosynthesis to natural and projected high seawater temperatures estimated by pulse amplitude modulation fluorometry in a temperate coral

Directory of Open Access Journals (Sweden)

Erik eCaroselli

2015-11-01

Full Text Available Balanophyllia europaea is a shallow water solitary zooxanthellate coral, endemic to the Mediterranean Sea. Extensive field studies across a latitudinal temperature gradient highlight detrimental effects of rising temperatures on its growth, demography and skeletal characteristics, suggesting that depression of photosynthesis at high temperatures might cause these negative effects. Here we test this hypothesis by analyzing, by means of pulse amplitude modulation fluorometry, the photosynthetic efficiency of B. europaea specimens exposed in aquaria to the annual range of temperatures experienced in the field (13°C, 18°C, and 28°C, and two extreme temperatures expected for 2100 as a consequence of global warming (29°C and 32°C. The indicators of photosynthetic performance analyzed (maximum and effective quantum yield showed that maximum efficiency was reached at 20.0-21.6°C, slightly higher than the annual mean temperature in the field (18°C. Photosynthetic efficiency decreased from 20.0°C to 13°C and even more strongly from 21.6°C to 32°C. An unusual form of bleaching was observed, with a maximum zooxanthellae density at 18°C that strongly decreased from 18°C to 32°C. Chlorophyll a concentration per zooxanthellae cell showed an opposite trend as it was minimal at 18°C and increased from 18°C to 32°C. Since the areal chlorophyll concentration is the product of the zooxanthellae density and its cellular content, these trends resulted in a homogeneous chlorophyll concentration per coral surface across temperature treatments. This confirms that B. europaea photosynthesis is progressively depressed at temperatures >21.6°C, supporting previous hypotheses raised by the studies on growth and demography of this species. This study also confirms the threats posed to this species by the ongoing seawater warming.

Integrating Biology into the General Chemistry Laboratory: Fluorometric Analysis of Chlorophyll "a"

Science.gov (United States)

Wesolowski, Meredith C.

2014-01-01

A laboratory experiment that introduces fluorometry of chlorophyll "a" at the general chemistry level is described. The use of thin-layer chromatography to isolate chlorophyll "a" from spirulina and leaf matter enables quantification of small amounts of chlorophyll "a" via fluorometry. Student results were reasonably…

Geochemistry and uranium content of phosphorites used in fertilizer industry - Homs in comparison with mines raw phosphorites

International Nuclear Information System (INIS)

Kudsi, Yahia; Abbas, Mahmoud

1992-03-01

geochemical characteristics of phosphates used in fertilizer industry at Homs were studied. 28 monthly averaged samples from Palmyrides mining areas were mineralogically and chemically analyzed for P 2 O 5 , CaO and U. Specifically, U content was determined by fluorometry, x-ray fluorescence and γ spectrometry. Results show that the phosphates under study are similar to that exploited at Charquieh mines. U shows a positive correlation with P 2 O 5 and not with CaO. Statistical analyses indicate that fluorometry is more accurate than x-ray fluorescence and γ spectrometry. (author). 14 refs., 23 figs., 5 tabs

Determination of uranium in urine by fluorometry

International Nuclear Information System (INIS)

Afsar, M.; Aziz, A.; Mubarak, M.A.

1978-01-01

The report describes fluorometric determination of traces of uranium in the urine of mine workers and persons handling uranium. After preliminary treatment uranium is extracted by solvent extraction with TBP:MIBK mixtures. An aliquot of the extracted solution is fused with high carbonate flux at 650 0 C in platinum dishes. The fluorescence of fused sample is measured using a fluorometer. Various fusion parameters are discussed. The procedure for the operation and calibration of fluorometer is also described

Toward autonomous measurements of photosynthetic electron transport rates: An evaluation of active fluorescence-based measurements of photochemistry

Czech Academy of Sciences Publication Activity Database

Silsbe, G.; Oxborough, K.; Suggett, D. J.; Forster, R. M.; Ihnken, S.; Komárek, Ondřej; Lawrenz, Evelyn; Prášil, Ondřej; Roettgers, R.; Šicner, M.; Simis, S. G. H.; Van Dijk, M. A.; Kromkamp, J. C.

2015-01-01

Roč. 13, MAR 2015 (2015), s. 138-155 ISSN 1541-5856 Institutional support: RVO:61388971 Keywords : REPETITION-RATE FLUOROMETRY * CAVITY ABSORPTION METER * CHLOROPHYLL FLUORESCENCE Subject RIV: EE - Microbiology, Virology Impact factor: 2.008, year: 2015

Divergent composition of algal-bacterial biofilms developing under various external factors

NARCIS (Netherlands)

Barranguet, C.; Veuger, B.; van Beusekom, S.A.M.; Marvan, P.; Sinke, J.J.; Admiraal, W.

2005-01-01

The influence of external factors other than nutrients on biofilm development and composition was studied with a combination of optical (Confocal Laser Scanning Microscopy, PAM fluorometry) and chemical methods (EPS extraction, HPLC, TOC determination). The development of algal-bacterial biofilms

J_'-9_7__ Degloving injuries and flap viability assesstnent

African Journals Online (AJOL)

S Air Med J 1993; 83: 97-99. TIe inevitable concomitant ... include surface fluorometry, ultrasound, laser Doppler, ... This is the cut-off area beyond which the dis- tal flap should .... groups according to weight loss measurements. The timing of ...

Time-resolved fluorometry of the aromatic amino acids

International Nuclear Information System (INIS)

Laws, W.R.; Ross, J.B.A.; Katsoyannis, P.G.; Wyssbrod, H.R.

1988-01-01

The aromatic amino acids tryptophan, tyrosine, and phenylalanine are the chromophores that produce the intrinsic fluorescence of proteins. It has been a long-desired goal to be able to use the fluorescence of these amino acids to help understand protein dynamics, structure, and function. Considerable information about proteins in solution has come from steady-state, or time-averaged, fluorescence measurements, especially from the fluorescence of tryptophan. For a protein to be characterized more extensively, however, the time dependence of the intrinsic fluorescence must also be explained. Unfortunately, complex fluorescence decay kinetics have been observed not only for proteins having just a single aromatic amino acid, but also for simple analogues of these amino acids; the cause of these complex decays is not fully understood. Considerable effort must still be made to resolve the mechanisms causing the complex decays

Chlorophyll in tomato seeds: marker for seed performance?

NARCIS (Netherlands)

Suhartanto, M.R.

2002-01-01

Using Xe-PAM, laser induced fluorometry and high performance liquid chromatography we found that chlorophyll was present in young tomato (cv. Moneymaker) seeds and was degraded during maturation. Fluorescence microscopy and imaging showed that the majority of chlorophyll is located in the

Magnitude of a conformational change in the glycine receptor beta1-beta2 loop is correlated with agonist efficacy

DEFF Research Database (Denmark)

Pless, Stephan Alexander; Lynch, Joseph W

2009-01-01

associated with the closed-flip transition in the alpha1-glycine receptor. We employed voltage-clamp fluorometry to compare ligand-binding domain conformational changes induced by the following agonists, listed from highest to lowest affinity and efficacy: glycine > beta-alanine > taurine. Voltage...

Molecular Basis for Allosteric Inhibition of Acid-Sensing Ion Channel 1a by Ibuprofen

DEFF Research Database (Denmark)

Lynagh, Timothy; Romero-Rojo, José Luis; Lund, Camilla

2017-01-01

-clamp fluorometry. Our results show that ibuprofen is an allosteric inhibitor of ASIC1a, which binds to a crucial site in the agonist transduction pathway and causes conformational changes that oppose channel activation. Ibuprofen inhibits several ASIC subtypes, but certain ibuprofen derivatives show some...

Quantitative Assessment of Spray Deposition with Water-Sensitive Paper

Science.gov (United States)

Spray droplets, discharged from the lower six nozzles of an airblast sprayer, were sampled on pairs of absorbent filter and water-sensitive papers at nine distances from sprayer. Spray deposition on filter targets were measured by fluorometry and spray distribution on WSP targets were assessed by t...

Comparative effects of herbicides on photosynthesis and growth of tropical estuarine microalgae

Energy Technology Data Exchange (ETDEWEB)

Magnusson, Marie [AIMS at JCU, Australian Institute of Marine Science, School of Marine and Tropical Biology, James Cook University, Townsville, QLD 4811 (Australia); School of Marine and Tropical Biology, James Cook University, Douglas Campus, Townsville 4811 (Australia); Heimann, Kirsten [School of Marine and Tropical Biology, James Cook University, Douglas Campus, Townsville 4811 (Australia)], E-mail: Kirsten.Heimann@jcu.edu.au; Negri, Andrew P. [Australian Institute of Marine Science, PMB 3 Townsville MC, QLD 4810 (Australia)

2008-09-15

Pulse amplitude modulation (PAM) fluorometry is ideally suited to measure the sub-lethal impacts of photosystem II (PSII)-inhibiting herbicides on microalgae, but key relationships between effective quantum yield [Y(II)] and the traditional endpoints growth rate ({mu}) and biomass increase are unknown. The effects of three PSII-inhibiting herbicides; diuron, hexazinone and atrazine, were examined on two tropical benthic microalgae; Navicula sp. (Heterokontophyta) and Nephroselmis pyriformis (Chlorophyta). The relationships between Y(II), {mu} and biomass increase were consistent (r{sup 2} {>=} 0.90) and linear (1:1), validating the utility of PAM fluorometry as a rapid and reliable technique to measure sub-lethal toxicity thresholds of PSII-inhibiting herbicides in these microalgae. The order of toxicity (EC{sub 50} range) was: diuron (16-33 nM) > hexazinone (25-110 nM) > atrazine (130-620 nm) for both algal species. Growth rate and photosynthesis were affected at diuron concentrations that have been detected in coastal areas of the Great Barrier Reef.

Comparative effects of herbicides on photosynthesis and growth of tropical estuarine microalgae

International Nuclear Information System (INIS)

Magnusson, Marie; Heimann, Kirsten; Negri, Andrew P.

2008-01-01

Pulse amplitude modulation (PAM) fluorometry is ideally suited to measure the sub-lethal impacts of photosystem II (PSII)-inhibiting herbicides on microalgae, but key relationships between effective quantum yield [Y(II)] and the traditional endpoints growth rate (μ) and biomass increase are unknown. The effects of three PSII-inhibiting herbicides; diuron, hexazinone and atrazine, were examined on two tropical benthic microalgae; Navicula sp. (Heterokontophyta) and Nephroselmis pyriformis (Chlorophyta). The relationships between Y(II), μ and biomass increase were consistent (r 2 ≥ 0.90) and linear (1:1), validating the utility of PAM fluorometry as a rapid and reliable technique to measure sub-lethal toxicity thresholds of PSII-inhibiting herbicides in these microalgae. The order of toxicity (EC 50 range) was: diuron (16-33 nM) > hexazinone (25-110 nM) > atrazine (130-620 nm) for both algal species. Growth rate and photosynthesis were affected at diuron concentrations that have been detected in coastal areas of the Great Barrier Reef

Structural rearrangement of the intracellular domains during AMPA receptor activation

DEFF Research Database (Denmark)

Zachariassen, Linda Grønborg; Katchan, Ljudmila; Jensen, Anna Guldvang

2016-01-01

-clamp fluorometry of the double- and single-insert constructs showed that both the intracellular C-terminal domain (CTD) and the loop region between the M1 and M2 helices move during activation and the CTD is detached from the membrane. Our time-resolved measurements revealed unexpectedly complex fluorescence...

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African Journals Online (AJOL)

be converted to fixed carbon. PAM fluorometry was ... rETRmax, α (initial slope before the onset of saturation),. Ik (minimum ... ference (HSD) analysis for comparison of means were carried out to test for ... were arcsin (square root) transformed prior to ANOVA analyses. ...... them to respond to fluctuating light regimes in differ-.

Fluorometric Analysis of Riboflavin.

Science.gov (United States)

Hu, Ji-Yue; And Others

1990-01-01

Described is the use of fluorometry in an experiment where the fluorescence of riboflavin is specifically quenched when titrated with riboflavin-binding protein. The amount of riboflavin, required by all organisms, in an unknown solution and the dissociation constant of the vitamin-protein complex can be determined using this method. (KR)

Lens fluorescence in relation to glucose tolerance and genetic predisposition to type 2 diabetes mellitus in a population-based study

DEFF Research Database (Denmark)

Koefoed Theil, Pernille; Kessel, Line; Hansen, Torben

2006-01-01

The fluorescence of the lens has the characteristics of a life-long cumulative index of glycemia, and it is elevated in patients with diabetes in proportion to the duration of diabetes and the level of glycemia. Consequently, lens fluorometry should be capable of providing an estimate of prediagn...... fluorescence values....

Frequency-domain lifetime fluorometry of double-labeled creatine kinase.

Science.gov (United States)

Gregor, M; Kubala, M; Amler, E; Mejsnar, J

2003-01-01

Myofibril-bound creatine kinase EC 2.7.3.2 (CK), a key enzyme of muscle energy metabolism, has been selected for studies of conformational changes that underlie the cellular control of enzyme activity. For fluorescence spectroscopy measurements, the CK molecule was double-labeled with IAF (5-iodoacetamidofluorescein) and ErITC (erythrosin 5'-isothiocyanate). Measurement of fluorescence resonance energy transfer (FRET) from fluorescein to erythrosin was used to obtain information about the donor-acceptor pair distance. Frequency-domain lifetime measurements evaluate the donor-acceptor distance in the native CK molecule as 7.8 nm. The Förster radius equals 5.3 nm with the resolution range from 0.2 to 1.0 nm. Erythrosin-fluorescein labeling (EFL) was tested for artificial conformational changes of the CK molecule with high-salt concentration treatment. The transition distance, defined by His-97 and Cys-283 and derived from a 3D model equals 0.766 nm for the open (inactive) form and 0.277 nm for the closed (reactive) form of the CK molecule. In this way, the resolution range of the used spectroscopy method is significant, concerning the difference of 0.489 nm. Nevertheless, the CK enzyme activity, assessed by the hexokinase-coupled assay, was diminished down to 1 % of the activity of the native enzyme. EFL is suitable for description of conformational behavior implied from the regulation of creatine kinase. However, the observed inhibition restricts EFL to studies of conformational changes during natural catalytic activity.

The measurement of natural uranium in urine by fluorometry

International Nuclear Information System (INIS)

Kramer, G.H.; Johnson, J.R.; Green, W.

1984-02-01

The fluorometric method of measuring natural uranium in urine that is currently used by the Bioassay Laboratory at Chalk River Nuclear Laboratories has been tested, optimized and documented. The method, which measures the fluorescence of uranium in a fused sodium fluoride pellet, has been shown to be quench independent and is routinely used to measure uranium concentrations in the range of 1 μg/L to 90 μg/L. The fluorimeter has a dynamic range of 0.2 μg/L to 200 μg/L

A method to decompose spectral changes in Synechocystis PCC 6803 during light-induced state transitions

Czech Academy of Sciences Publication Activity Database

Acuna, A.M.; Kaňa, Radek; Gwizdala, M.; Snellenburg, J.J.; van Alphen, P.; van Oort, B.; Kirilovsky, D.; van Grondelle, R.; van Stokkum, I.H.M.

2016-01-01

Roč. 130, 1-3 SI (2016), s. 237-249 ISSN 0166-8595 R&D Projects: GA ČR GBP501/12/G055; GA MŠk(CZ) LO1416; GA MŠk(CZ) ED2.1.00/19.0392 Institutional support: RVO:61388971 Keywords : Cyanobacteria * Spectrally resolved fluorometry * Singular value decomposition Subject RIV: EF - Botanics Impact factor: 3.864, year: 2016

In situ Measurements of Phytoplankton Fluorescence Using Low Cost Electronics

Directory of Open Access Journals (Sweden)

Dana L. Wright

2013-06-01

Full Text Available Chlorophyll a fluorometry has long been used as a method to study phytoplankton in the ocean. In situ fluorometry is used frequently in oceanography to provide depth-resolved estimates of phytoplankton biomass. However, the high price of commercially manufactured in situ fluorometers has made them unavailable to some individuals and institutions. Presented here is an investigation into building an in situ fluorometer using low cost electronics. The goal was to construct an easily reproducible in situ fluorometer from simple and widely available electronic components. The simplicity and modest cost of the sensor makes it valuable to students and professionals alike. Open source sharing of architecture and software will allow students to reconstruct and customize the sensor on a small budget. Research applications that require numerous in situ fluorometers or expendable fluorometers can also benefit from this study. The sensor costs US$150.00 and can be constructed with little to no previous experience. The sensor uses a blue LED to excite chlorophyll a and measures fluorescence using a silicon photodiode. The sensor is controlled by an Arduino microcontroller that also serves as a data logger.

Pulse amplitude modulated chlorophyll fluorometer

Science.gov (United States)

Greenbaum, Elias; Wu, Jie

2015-12-29

Chlorophyll fluorometry may be used for detecting toxins in a sample because of changes in micro algae. A portable lab on a chip ("LOAC") based chlorophyll fluorometer may be used for toxin detection and environmental monitoring. In particular, the system may include a microfluidic pulse amplitude modulated ("PAM") chlorophyll fluorometer. The LOAC PAM chlorophyll fluorometer may analyze microalgae and cyanobacteria that grow naturally in source drinking water.

Leaching of uranium from Syrian phosphorite (sodium carbonate-bicarbonate)

International Nuclear Information System (INIS)

Abou-Jamous, J.Kh.

1991-01-01

The leaching of uranium from Syrian phosphorite by sodium carbonate-bicarbonate solution has been studied, using a batch technique. Parameters influencing percentage extraction of uranium that are considered and studies in this work are: Leachant concentration, particle size, heat treatment, leachant renewal, phosphorite renewal and contact time. All measurements of uranium from aqueous solutions were carried out by fluorometry. (author). 12 refs., 4 figs., 1 tab

Urinary extracellular vesicles for RNA extraction: optimization of a protocol devoid of prokaryote contamination.

Science.gov (United States)

Tataruch-Weinert, Dorota; Musante, Luca; Kretz, Oliver; Holthofer, Harry

2016-01-01

Urinary extracellular vesicles (UEVs) represent an ideal platform for biomarker discovery. They carry different types of RNA species, and reported profile discrepancies related to the presence/absence of 18s and 28s rRNA remain controversial. Moreover, sufficient urinary RNA yields and respective quality RNA profiles are still to be fully established. UEVs were enriched by hydrostatic filtration dialysis, and RNA content was extracted using 7 different commercially available techniques. RNA quantity was assessed using spectrophotometry and fluorometry, whilst RNA quality was determined by capillary electrophoresis. The presence of prokaryotic transcriptome was stressed when cellular RNA, as a control, was spiked into the UEVs samples before RNA extraction. The presence of bacteria in hydrostatic filtration dialysis above 1,000 kDa molecular weight cut-off and in crude urine was confirmed with growth media plates. The efficiency in removing urinary bacteria was evaluated by differential centrifugation, filtration (0.22 µm filters) and chemical pretreatment (water purification tablet). For volumes of urine >200 ml, the chemical treatment provides ease of handling without affecting vesicle integrity, protein and RNA profiles. This protocol was selected to enrich RNA with 7 methods, and its respective quality and quantity were assessed. The results were given as follows: (a) Fluorometry gave more repeatability and reproducibility than spectrophotometry to assess the RNA yields, (b) UEVs were enriched with small RNA, (c) Ribosomal RNA peaks were not observed for any RNA extraction method used and (d) RNA yield was higher for column-based method designed for urinary exosome, whilst the highest relative microRNA presence was obtained using TRIzol method. Our results show that the presence of bacteria can lead to misidentification in the electrophoresis peaks. Fluorometry is more reliable than spectrophotometry. RNA isolation method must be selected in conjunction with

Urinary extracellular vesicles for RNA extraction: optimization of a protocol devoid of prokaryote contamination

Directory of Open Access Journals (Sweden)

Dorota Tataruch-Weinert

2016-06-01

Full Text Available Background: Urinary extracellular vesicles (UEVs represent an ideal platform for biomarker discovery. They carry different types of RNA species, and reported profile discrepancies related to the presence/absence of 18s and 28s rRNA remain controversial. Moreover, sufficient urinary RNA yields and respective quality RNA profiles are still to be fully established. Methods: UEVs were enriched by hydrostatic filtration dialysis, and RNA content was extracted using 7 different commercially available techniques. RNA quantity was assessed using spectrophotometry and fluorometry, whilst RNA quality was determined by capillary electrophoresis. Results: The presence of prokaryotic transcriptome was stressed when cellular RNA, as a control, was spiked into the UEVs samples before RNA extraction. The presence of bacteria in hydrostatic filtration dialysis above 1,000 kDa molecular weight cut-off and in crude urine was confirmed with growth media plates. The efficiency in removing urinary bacteria was evaluated by differential centrifugation, filtration (0.22 µm filters and chemical pretreatment (water purification tablet. For volumes of urine >200 ml, the chemical treatment provides ease of handling without affecting vesicle integrity, protein and RNA profiles. This protocol was selected to enrich RNA with 7 methods, and its respective quality and quantity were assessed. The results were given as follows: (a Fluorometry gave more repeatability and reproducibility than spectrophotometry to assess the RNA yields, (b UEVs were enriched with small RNA, (c Ribosomal RNA peaks were not observed for any RNA extraction method used and (d RNA yield was higher for column-based method designed for urinary exosome, whilst the highest relative microRNA presence was obtained using TRIzol method. Conclusion: Our results show that the presence of bacteria can lead to misidentification in the electrophoresis peaks. Fluorometry is more reliable than spectrophotometry. RNA

The determination of traces of uranium by means of laser fluorometry

International Nuclear Information System (INIS)

Venter, R.J.

1987-11-01

A method and apparatus were developed to quantitatively determine traces of uranium. A xenon chloride excimer laser was used as an excitation source, which provided various advantages above the conventional light sources. Boxcar integration was used to limit background fluorescence by time-resolved fluorescence spectroscopy. A characteristic wavelength of uranium fluorescence was selected with cut-off filters and a monochromator. The effect of different matrices on the fluorescence of uranium was determined, and the most suitable matrix for quantitative analysis was chosen. A quick and simple method for determining uranium was developed. A combined water sample of industrial effluent was used to evaluate the method. The effect of interfering species, characteristic of the water sample, was examined. A standard addition technique was used to compensate for the effect of the interfering species. The detection limit of uranium in pure water was calculated at three times the standard deviation of the blank sample and was found to be 14 pg/cm 3

Gene expression of a green fluorescent protein homolog as a host-specific biomarker of heat stress within a reef-building coral.

Science.gov (United States)

Smith-Keune, C; Dove, S

2008-01-01

Recent incidences of mass coral bleaching indicate that major reef building corals are increasingly suffering thermal stress associated with climate-related temperature increases. The development of pulse amplitude modulated (PAM) fluorometry has enabled rapid detection of the onset of thermal stress within coral algal symbionts, but sensitive biomarkers of thermal stress specific to the host coral have been slower to emerge. Differential display reverse transcription polymerase chain reaction (DDRT-PCR) was used to produce fingerprints of gene expression for the reef-building coral Acropora millepora exposed to 33 degrees C. Changes in the expression of 23 out of 399 putative genes occurred within 144 h. Down-regulation of one host-specific gene (AmA1a) occurred within just 6 h. Full-length sequencing revealed the product of this gene to be an all-protein chromatophore (green fluorescent protein [GFP]-homolog). RT-PCR revealed consistent down-regulation of this GFP-homolog for three replicate colonies within 6 h at both 32 degrees C and 33 degrees C but not at lower temperatures. Down-regulation of this host gene preceded significant decreases in the photosynthetic activity of photosystem II (dark-adapted F (v)/F (m)) of algal symbionts as measured by PAM fluorometry. Gene expression of host-specific genes such as GFP-homologs may therefore prove to be highly sensitive indicators for the onset of thermal stress within host coral cells.

Contributions of counter-charge in a potassium channel voltage-sensor domain

DEFF Research Database (Denmark)

Pless, Stephan Alexander; Galpin, Jason D; Niciforovic, Ana P

2011-01-01

Voltage-sensor domains couple membrane potential to conformational changes in voltage-gated ion channels and phosphatases. Highly coevolved acidic and aromatic side chains assist the transfer of cationic side chains across the transmembrane electric field during voltage sensing. We investigated...... the functional contribution of negative electrostatic potentials from these residues to channel gating and voltage sensing with unnatural amino acid mutagenesis, electrophysiology, voltage-clamp fluorometry and ab initio calculations. The data show that neutralization of two conserved acidic side chains...

Radiometric parameters in freshwater samples of Centro Experimental Aramar (CTMSP/Brazil)

Energy Technology Data Exchange (ETDEWEB)

Moraes, Marco Antonio P.V. [Laboratorio Radioecologico (LARE), Ipero, SP (Brazil). Dept. de Seguranca Nuclear; Instituto de Pesquisas Energeticas e Nucleares (IPEN/CNEN-SP), Sao Paulo, SP (Brazil)

2011-07-01

Experimental results are described obtained with gamma spectrometry, alpha and beta gross counts, liquid scintillation and fluorometry techniques for the measurement of background radiation in surface water samples, collected in Centro Experimental Aramar and surroundings, from 1988 to 2009. The estimated average background radiation concentrations in water samples in this region are low, related to the low level detection limits of the techniques, and indicates good water quality parameters, and low interference in the environment in Centro Experimental Aramar and region. (author)

Lead Poison Detection

Science.gov (United States)

1976-01-01

With NASA contracts, Whittaker Corporations Space Science division has developed an electro-optical instrument to mass screen for lead poisoning. Device is portable and detects protoporphyrin in whole blood. Free corpuscular porphyrins occur as an early effect of lead ingestion. Also detects lead in urine used to confirm blood tests. Test is inexpensive and can be applied by relatively unskilled personnel. Similar Whittaker fluorometry device called "drug screen" can measure morphine and quinine in urine much faster and cheaper than other methods.

Herbicide toxicity on river biofilms assessed by pulse amplitude modulated (PAM) fluorometry

International Nuclear Information System (INIS)

Kim Tiam, Sandra; Laviale, Martin; Feurtet-Mazel, Agnès; Jan, Gwilherm; Gonzalez, Patrice; Mazzella, Nicolas; Morin, Soizic

2015-01-01

Highlights: • Rapid Light Curves were shown to be early markers of toxicant exposure. • Diuron and norflurazon effects were significant at environmentally realistic concentrations. • Toxic effects in intact biofilms seem to be delayed compared to disrupted biofilms. - Abstract: The use of Rapid light curves (RLCs) as a toxicity endpoint for river biofilms was examined in this study and compared to “classical fluorescence parameters” i.e. minimal fluorescence (F 0 ), optimal and effective quantum yields of photosystem II (F v /F m and Φ PSII ). Measurements were performed after exposure to five concentrations of diuron (from 0.3 to 33.4 μg L −1 ), its main degradation product (DCPMU) (from 1.0 to 1014 μg L −1 ) and norflurazon (from 0.6 to 585 μg L −1 ) with the lowest exposure concentrations corresponding to levels regularly encountered in chronically contaminated sites. Biofilm responses were evaluated after 1, 5, 7 and 14 days of exposure to the different toxicants. Overall, the responses of both “classical fluorescence parameters” and RLC endpoints were highly time dependent and related to the mode of action of the different compounds. Interestingly, parameters calculated from RLCs (α, ETR max and I k ) were useful early markers of pesticide exposure since they revealed significant effects of all the tested toxicants from the first day of exposure. In comparison, classical fluorescence endpoints (F 0 and F v /F m ) measured at day 1 were only affected in the DCPMU treatment. Our results demonstrated the interest of RLCs as early markers of toxicant exposure particularly when working with toxicants with less specific mode of action than PSII inhibitors

Herbicide toxicity on river biofilms assessed by pulse amplitude modulated (PAM) fluorometry

Energy Technology Data Exchange (ETDEWEB)

Kim Tiam, Sandra, E-mail: sandra.kimtiam@gmail.com [Irstea, UR EABX, 50 Avenue de Verdun, F-33612, Cestas Cedex (France); Université de Bordeaux, EPOC, UMR 5805, F-33120 Arcachon (France); Laviale, Martin [Departamento de Biologia and CESAM – Centro de Estudos do Ambiente e do Mar Universidade de Aveiro, Campus de Santiago, 3810-193 Aveiro (Portugal); Sorbonne Universités, UPMC Univ Paris 06, UMR 7093, LOV, Observatoire Océanologique, F-06230, Villefranche-Sur-Mer (France); CNRS, UMR 7093, LOV, Observatoire Océanologique, F-06230, Villefranche-Sur-Mer France (France); Feurtet-Mazel, Agnès [Université de Bordeaux, EPOC, UMR 5805, F-33120 Arcachon (France); Jan, Gwilherm [Irstea, UR EABX, 50 Avenue de Verdun, F-33612, Cestas Cedex (France); Gonzalez, Patrice [Université de Bordeaux, EPOC, UMR 5805, F-33120 Arcachon (France); Mazzella, Nicolas; Morin, Soizic [Irstea, UR EABX, 50 Avenue de Verdun, F-33612, Cestas Cedex (France)

2015-08-15

Highlights: • Rapid Light Curves were shown to be early markers of toxicant exposure. • Diuron and norflurazon effects were significant at environmentally realistic concentrations. • Toxic effects in intact biofilms seem to be delayed compared to disrupted biofilms. - Abstract: The use of Rapid light curves (RLCs) as a toxicity endpoint for river biofilms was examined in this study and compared to “classical fluorescence parameters” i.e. minimal fluorescence (F{sub 0}), optimal and effective quantum yields of photosystem II (F{sub v}/F{sub m} and Φ{sub PSII}). Measurements were performed after exposure to five concentrations of diuron (from 0.3 to 33.4 μg L{sup −1}), its main degradation product (DCPMU) (from 1.0 to 1014 μg L{sup −1}) and norflurazon (from 0.6 to 585 μg L{sup −1}) with the lowest exposure concentrations corresponding to levels regularly encountered in chronically contaminated sites. Biofilm responses were evaluated after 1, 5, 7 and 14 days of exposure to the different toxicants. Overall, the responses of both “classical fluorescence parameters” and RLC endpoints were highly time dependent and related to the mode of action of the different compounds. Interestingly, parameters calculated from RLCs (α, ETR{sub max} and I{sub k}) were useful early markers of pesticide exposure since they revealed significant effects of all the tested toxicants from the first day of exposure. In comparison, classical fluorescence endpoints (F{sub 0} and F{sub v}/F{sub m}) measured at day 1 were only affected in the DCPMU treatment. Our results demonstrated the interest of RLCs as early markers of toxicant exposure particularly when working with toxicants with less specific mode of action than PSII inhibitors.

Fluorescence decay time imaging using an imaging photon detector with a radio frequency photon correlation system

Science.gov (United States)

Morgan, Christopher G.; Mitchell, A. C.; Murray, J. G.

1990-05-01

An imaging photon detector has been modified to incorporate fast timing electronics coupled to a custom built photon correlator interfaced to a RISC computer. Using excitation with intensity- muodulated light, fluorescence images can be readily obtained where contrast is determined by the decay time of emission, rather than by intensity. This technology is readily extended to multifrequency phase/demodulation fluorescence imaging or to differential polarised phase fluorometry. The potential use of the correlator for confocal imaging with a laser scanner is also briefly discussed.

Investigation of cell-free DNA in canine plasma and its relation to disease.

Science.gov (United States)

Burnett, Deborah L; Cave, Nicholas J; Gedye, Kristene R; Bridges, Janis P

2016-09-01

DNA is released from dying cells during apoptosis and necrosis. This cell-free DNA (cfDNA) diffuses into the plasma where it can be measured. In humans, an increase in cfDNA correlates with disease severity and prognosis. It was hypothesized that when DNA in canine plasma was measured by emission fluorometry without prior DNA extraction, the concentration of cfDNA would increase with disease severity. The diseased population consisted of 97 client-owned dogs. The clinically normal population consisted of nine client-owned dogs presenting for 'wellness screens', and 15 colony-owned Harrier Hounds. Plasma cfDNA was measured by fluorometry without prior DNA extraction. The effects of ex vivo storage conditions were evaluated in plasma from two clinically normal dogs. In all other dogs, plasma was separated within two hours of collection. The association between the cfDNA concentration in hospitalized dogs and a variety of clinical, clinicopathological and outcome variables was tested. The concentration of cfDNA was reliably measured when plasma was separated within two hours of blood collection. The diseased dogs had significantly higher cfDNA than clinically normal dogs (P Dogs that did not survive to discharge had significantly higher cfDNA concentrations than survivors (P = 0.02). Conclusions/Clinical Importance: The concentration of cfDNA in the plasma of diseased dogs is associated with disease severity and prognosis. Measurement of canine cfDNA could be a useful non-specific disease indicator and prognostic tool.

Contrasting Photophysiological Characteristics of Phytoplankton Assemblages in the Northern South China Sea.

Science.gov (United States)

Jin, Peng; Gao, Guang; Liu, Xin; Li, Futian; Tong, Shanying; Ding, Jiancheng; Zhong, Zhihai; Liu, Nana; Gao, Kunshan

2016-01-01

The growth of phytoplankton and thus marine primary productivity depend on photophysiological performance of phytoplankton cells that respond to changing environmental conditions. The South China Sea (SCS) is the largest marginal sea of the western Pacific and plays important roles in modulating regional climate and carbon budget. However, little has been documented on photophysiological characteristics of phytoplankton in the SCS. For the first time, we investigated photophysiological characteristics of phytoplankton assemblages in the northern South China Sea (NSCS) using a real-time in-situ active chlorophyll a fluorometry, covering 4.0 × 105 km2. The functional absorption cross section of photosystem II (PSII) in darkness (σPSII) or under ambient light (σPSII') (A2 quanta-1) increased from the surface to deeper waters at all the stations during the survey period (29 July to 23 August 2012). While the maximum (Fv/Fm, measured in darkness) or effective (Fq'/Fm', measured under ambient light) photochemical efficiency of PSII appeared to increase with increasing depth at most stations, it showed inverse relationship with depth in river plume areas. The functional absorption cross section of PSII changes could be attributed to light-adapted genotypic feature due to niche-partition and the alteration of photochemical efficiency of PSII could be attributed to photo-acclimation. The chlorophyll a fluorometry can be taken as an analog to estimate primary productivity, since areas of higher photochemical efficiency of PSII coincided with those of higher primary productivity reported previously in the NSCS.

Health surveillance of personnel engaged in decontamination of depleted uranium contaminated regions

Energy Technology Data Exchange (ETDEWEB)

Djurovic, B. [Military Medical Academy, Radiological Protection Dept., Belgrade, Serbia and Montenegro (Yugoslavia); Spasic-Jokic, V. [ESLA Accelerator Installation, Lab. of Physics, VINCA Institute of Nuclear Sciences, Belgrade, Serbia and Montenegro (Yugoslavia); Fortuna, D.; Milenkovic, M. [NBH Military Educational Center, Krusevac, Serbia and Montenegro (Yugoslavia)

2006-07-01

After the NATO actions against Serbia and Montenegro, 112 locations were highly contaminated with depleted uranium-112 locations in Kosovo, 7 in the south of Serbia and 1 in Montenegro. Contaminated regions were marked, isolated and some of them decontaminated. In this paper we present the health surveillance protocol created for personnel engaged in decontamination of contaminated regions of Pljackovica and Bratoselce. They were examined and selected before decontamination and only healthy professionals (36 and 28) were engaged. Examination included: general clinical assessment, complete blood count with differential white blood cells; biochemical analysis of blood and urine, specifically renal and liver functions tests, cytogenetic tests (chromosomal aberration and micronucleus test), and laser fluorometry of 24-h urine sample and gamma spectrometry of the same if the levels were elevated. After the decontamination in the first group no clinical or biochemical changes were found, but in 3 of 36 were found unstable chromosomal aberrations. In the second group, in 3 of 28 were found unstable chromosomal aberrations and in 3 of them laser fluorometry analysis showed elevated levels of uranium (>3 {mu}g/l in two, and >5 {mu}g/l in one of them). Gamma spectrometry showed that it was not depleted, but naturally occurring uranium. Additionally performed analysis showed they were from the same village which is in the zone of highly elevated uranium level in ground and water. Three months later no chromosomal changes were found. (authors)

Evaluation of laser phosphorimetry for the analysis of uranium in biological samples from laboratory animal studies

International Nuclear Information System (INIS)

Gray, D.; Eidsom, A.F.

1985-01-01

Laser phosphorimetry has been used for uranium analyses in a variety of sample matrices, including environmental and human bioassay samples. The Scientrex-UA-3 Uranium Analyzer has been used at ITRI to acquire data on the applicability of laser phosphorimetry to analyses of uranium in the highly concentrated solutions resulting from chemical processing of biological comparisons of results with those obtained from conventional fluorometry. These comparisons have been very favorable for many sample types. Results of these comparisons and an evaluation of the data obtained with the Scintrex unit are presented

[Determination of polysaccharide from Chinese medicine Morinda officinalis how and its trace elements analysis].

Science.gov (United States)

Wu, Yong-Jun; Liu, Jie; Wu, Yu-Ming; Liu, Li-E; Zhang, Hong-Quan

2005-12-01

Polysaccharide was extracted from morinda officinalis how by back-flowing with 80% ethanol for 1 h, and the method of improved phenol-sulfuric acid spectroscopy was adopted to determine the content of morinda officinalis how polysaccharide. As for its trace elements, the atomic absorption spectroscopy was used to detect the content of Zn, Fe and Cu, which were compared with its fake; and cold atomic fluorometry was applied to determine the trace mercury. The results were satisfactory, which can give reference about the effective components of morinda officinalis how, and will help to exploit it.

Diverse and Tissue Specific Mitochondrial Respiratory Response in A Mouse Model of Sepsis-Induced Multiple Organ Failure

DEFF Research Database (Denmark)

Karlsson, Michael; Hara, Naomi; Morata, Saori

2016-01-01

control ratio was also significantly increased. Maximal Protonophore-induced respiratory (uncoupled) capacity was similar between the two treatment groups.The present study suggests a diverse and tissue specific mitochondrial respiratory response to sepsis. The brain displayed an early impaired...... C57BL/6 mice were analyzed at either 6 hours or 24 hours. ROS-production was simultaneously measured in brain samples using fluorometry.Septic brain tissue exhibited an early increased uncoupling of respiration. Temporal changes between the two time points were diminutive and no difference in ROS...

17β-estradiol induces non-genomic effects in renal intercalated cells through the G-protein coupled estrogen receptor 1

DEFF Research Database (Denmark)

Hofmeister, Marlene Vind; Damkier, Helle Hasager; Christensen, Birgitte Mønster

2012-01-01

Steroid hormones such as 17β-estradiol (E2) are known to modulate ion transporter expression in the kidney through classic intracellular receptors. Steroid hormones are also known to cause rapid nongenomic responses in a variety of nonrenal tissues. However, little is known about renal short......-term effects of steroid hormones. Here, we studied the acute actions of E2 on intracellular Ca(2+) signaling in isolated distal convoluted tubules (DCT2), connecting tubules (CNT), and initial cortical collecting ducts (iCCD) by fluo 4 fluorometry. Physiological concentrations of E2 induced transient increases...

Fluorescence properties of europium and samarium. beta. -diketonates and their use in fluorometry

Energy Technology Data Exchange (ETDEWEB)

Huang, H; Hiraki, K; Nishikawa, Y [Kinki Univ., Higashi-Osaka, Osaka (Japan). Faculty of Science and Technology

1981-01-01

Several europium and samarium ..beta..-diketonates (tta, ntfa, bfa) complexed with 1, 10-phenanthroline, or with trioctylphosphine oxide (topo) were synthesized. The fluorescence properties of these compounds in benzene or hexane have been studied. Absorption and fluorescence spectra, fluorescence quantum yield, fluorescence sensitivity index (F.S.I.), and fluorescence lifetime were measured. From the measurement of fluorescence lifetime of the ..beta..-diketonates, the velocity of radiative process (k sub(f)/phi sub(f)) has almost the same value for benzene and hexane solvent. The red fluorescence (Em. max. : 619 nm) of Eu(III) in these chelates is attributed to transitions from /sup 5/D/sub 0/ ..-->.. /sup 7/F/sub 2/ levels of this ion, and the three-band spectrum (Em. max. : 569 nm, 606 nm, 650 nm) indicates the transitions from the /sup 4/G sub(5/2) ..-->.. /sup 6/H sub(5/2), /sup 4/G sub(5/2) ..-->.. /sup 6/H sub(7/2), and /sup 4/G sub(5/2) ..-->.. /sup 6/H sub(9/2) levels of Sm(III), respectively. These spectra are not changed by any solvents and ligands. From the results, the fluorescence of the ..beta..-diketonates in organic solvent has been attributed to m* ..-->.. m luminescence transition. The complexes of Eu(III) and Sm(III) show radiative transition within orbitals, composed exclusively of 4f orbitals of rare earth ions (m* ..-->.. m radiative transition). Fluorinated ligands show better sensitivity than unfluorinated ligands, and the best sensitivity is obtained with TTA-phen system, and/or TTA-topo system for the spectrofluorometric determination of the two metals. In the case of Eu determination, 619 nm emission wavelength is used (the determinable range : 0.2 -- 10 ppb Eu), and in the case of Sm determination, 650 nm emission wavelength is adopted (the determinable range : 0.1 -- 1 ppm Sm), because of much higher sensitivity than the other two peaks (569, 606 nm) without interference from europium complex.

On the use of rhodamine WT for the characterization of stream hydrodynamics and transient storage

Science.gov (United States)

Runkel, Robert L.

2015-01-01

Recent advances in fluorometry have led to increased use of rhodamine WT as a tracer in streams and rivers. In light of this increased use, a review of the dye's behavior in freshwater systems is presented. Studies in the groundwater literature indicate that rhodamine WT is transported nonconservatively, with sorption removing substantial amounts of tracer mass. Column studies document a two-step breakthrough curve in which two structural isomers are chromatographically separated. Although the potential for nonconservative transport is acknowledged in the surface water literature, many studies assume that sorptive losses will not affect the characterization of physical transport processes. A literature review and modeling analysis indicates that this assumption is valid for quantification of physical properties that are based on the bulk of the tracer mass (traveltime), and invalid for the characterization of processes represented by the tracer tail (transient storage attributable to hyporheic exchange). Rhodamine WT should be considered nonconservative in the hyporheic zone due to nonconservative behavior demonstrated for similar conditions in groundwater. As such, rhodamine WT should not be used as a quantitative tracer in hyporheic zone investigations, including the study of long flow paths and the development of models describing hyporheic zone processes. Rhodamine WT may be used to qualitatively characterize storage in large systems, where there are few practical alternatives. Qualitative investigations should rely on early portions of the tracer profile, making use of the temporal resolution afforded by in situ fluorometry, while discarding later parts of the tracer profile that are adversely affected by sorption.

Enhanced immunoassay for porcine circovirus type 2 antibody using enzyme-loaded and quantum dots-embedded shell–core silica nanospheres based on enzyme-linked immunosorbent assay

International Nuclear Information System (INIS)

Wu, Long; Li, Xuepu; Shao, Kang; Ye, Shiyi; Liu, Chen; Zhang, Chenjun; Han, Heyou

2015-01-01

Boosting the detection sensitivity of enzyme-linked immunosorbent assay (ELISA) is significant to the early clinical diagnosis of various diseases. Here, we developed a versatile immunosensor using silica nanospheres as carriers for sensitive detection of porcine circovirus type 2 (PCV2) antibody. With HRP enzyme covalently immobilized on the silica nanospheres and CdSe nanocrystals embedded inside, these signal probes were successfully utilized in the sensitive detection of PCV2 antibody by ELISA, fluorometry and square-wave voltammetry (SWV). To further demonstrate the performance of the immunosensor, Human IgG (HIgG) was used as a model analyte. Since more HRP and CdSe QDs were loaded, 5-, 200- and 400-fold enhancements in amplified ELISA, fluorometry and voltammetry responses for HIgG could be achieved compared to conventional ELISA. The respective detection limits of theses methods for HIgG were 3.9, 0.1 and 0.05 ng mL −1 with a RSD below 5% for amplified ELISA, fluorescence and SWV measurements. Additionally, a 100-fold improvement was obtained in the detection sensitivity for PCV2 antibody immunoassay. The versatile immunosensor exhibits good sensitivity, stability and reproducibility, suggesting its potential applications in clinical diagnostics. - Highlights: • A versatile ELISA-based immunoassay for PCV2 antibody was developed. • Enzyme and CdSe QDs modified SiO 2 particles were used to improve sensitivity. • The simultaneous three ELISA-based techniques enhanced the detection reliability. • The biosensors strategy could provide a new avenue to ELISA-based sensors

Modification of HSP proteins and Ca2+ are responsible for the NO-derived peroxynitrite mediated neurological damage in PC12 cell.

Science.gov (United States)

Wen, Jun; Li, Hua; Zhang, Yudan; Li, Xia; Liu, Fang

2015-01-01

Peroxynitrite as one crucial metabolite of NO-derived agents has been well multi-investigated to inspect its potential role and sought to define its concrete mechanism underlying the memory loss and impaired cognition involved in pathological processes. In this investigation, the cell viability was assessed by the MTT assay. The neurotoxicity of peroxynitrite was analyzed by using immunohistochemical measurements in cultured PC12 cells to explore the underlying mechanisms. The generation of ROS was evaluated by a fluorometry assay by a fluorometry assay. Apoptosis was assayed by annexin V-FITC and PI staining with flow cytometry. [Ca2+]i was examined by using the microspectrofluorometer. Hsp70 was detected by western blot assay. The results revealed that PC12 cells were inhibited by peroxynitrite both in a dose-dependent and time-dependent manner. The level of ROS in PC12 cells exposed to SIN-1 was increased in a dose-dependent manner. The result indicated that the SIN-1 induced apoptosis of PC12 cells in a dose-dependent manner. Quercetin inhibited the viability of PC12 cells in a concentration-dependent manner. [Ca2+]i was increased gradually when cells treated with quercetin alone and also increased with treatment of dantrolene-containing. Hsp70 was significantly decreased in SIN-1-treated group compared with that of control group (P<0.01). In conclusion, Ca2+ homeostasis and chaperone Hsp70 were critically involved in peroxynitrite induced nitrosative stress as protective. Peroxynitrite acts as the pathological agent in learning and memory defects in CNS disorders associated with challenge.

Enhanced immunoassay for porcine circovirus type 2 antibody using enzyme-loaded and quantum dots-embedded shell–core silica nanospheres based on enzyme-linked immunosorbent assay

Energy Technology Data Exchange (ETDEWEB)

Wu, Long; Li, Xuepu; Shao, Kang; Ye, Shiyi; Liu, Chen; Zhang, Chenjun; Han, Heyou, E-mail: hyhan@mail.hzau.edu.cn

2015-08-05

Boosting the detection sensitivity of enzyme-linked immunosorbent assay (ELISA) is significant to the early clinical diagnosis of various diseases. Here, we developed a versatile immunosensor using silica nanospheres as carriers for sensitive detection of porcine circovirus type 2 (PCV2) antibody. With HRP enzyme covalently immobilized on the silica nanospheres and CdSe nanocrystals embedded inside, these signal probes were successfully utilized in the sensitive detection of PCV2 antibody by ELISA, fluorometry and square-wave voltammetry (SWV). To further demonstrate the performance of the immunosensor, Human IgG (HIgG) was used as a model analyte. Since more HRP and CdSe QDs were loaded, 5-, 200- and 400-fold enhancements in amplified ELISA, fluorometry and voltammetry responses for HIgG could be achieved compared to conventional ELISA. The respective detection limits of theses methods for HIgG were 3.9, 0.1 and 0.05 ng mL{sup −1} with a RSD below 5% for amplified ELISA, fluorescence and SWV measurements. Additionally, a 100-fold improvement was obtained in the detection sensitivity for PCV2 antibody immunoassay. The versatile immunosensor exhibits good sensitivity, stability and reproducibility, suggesting its potential applications in clinical diagnostics. - Highlights: • A versatile ELISA-based immunoassay for PCV2 antibody was developed. • Enzyme and CdSe QDs modified SiO{sub 2} particles were used to improve sensitivity. • The simultaneous three ELISA-based techniques enhanced the detection reliability. • The biosensors strategy could provide a new avenue to ELISA-based sensors.

Studies of cellular damage induced by X-rays and visible light

International Nuclear Information System (INIS)

Christensen, T.; Kinn, G.; Reitan, J.B.

1989-01-01

DNA-damage in cells has been studied by use of spectrophotometry and fluorometry. The method is based on the differential fluorescence quantum yield of the fluorochrome Hoechst 33258 when bound to single and double stranded DNA, respectively. DNA-damage by doses of X-rays below 2 Gy was clearly detectable. Blue light from phototherapy lamps induced DNA-damage in human TMG-1 glioblastoma, but no significant effect could be observed after irradiation with green lamps. In the presence of bilirubin the amount of DNA-damage was increased, notably at high bilirubin concentration and by blue light. 9 refs; 12 figs

Experimental evaluation of the detection threshold of uranium in urine samples

International Nuclear Information System (INIS)

Ferreyra, M. D.; Suarez Mendez, Sebastian; Tossi, Mirta H.

1999-01-01

The routine internal dosimetric tests for nuclear installations workers includes the determination of uranium in urine. The analysis is carried out, after chemical treatment, by UV fluorometry, comparing the results with urine blank samples from workers not exposed professionally to contamination. The fluctuation of the results of the uranium content in the blank samples greatly affects the determinations. In 30 blank samples the uranium content was determined and the results were evaluated by three calculation methods: 1) The procedure recommended by IUPAC; 2) The graphical method; 3) and The error propagation method. The last one has been adopted for the calculation of the detection threshold. (authors)

Fish stocking density impacts tank hydrodynamics

DEFF Research Database (Denmark)

Rasmussen, Michael R.; Lunger, Angela; Laursen, Jesper

2006-01-01

The effect of stocking density upon the hydrodynamics of a circular tank, configured in a recirculation system, was investigated. Red drums Sciaenops ocellatus of approximately 140 g wet weight, were stocked at five rates varying from 0 to 12 kg m-3. The impact of the presence of fish upon tank...... hydrodynamics was established using in-tank-based Rhodamine WT fluorometry at a flow rate of 0.23 l s-1 (tank exchange rate of 1.9 h-1). With increasing numbers of animals, curvilinear relationships were observed for dispersion coefficients and tank mixing times. Stocking densities of 3, 6, 9 and 12 kg m-3...

Identification of potential internal control genes for real-time PCR analysis during stress response in Pyropia haitanensis

Science.gov (United States)

Wang, Xia; Feng, Jianhua; Huang, Aiyou; He, Linwen; Niu, Jianfeng; Wang, Guangce

2017-11-01

Pyropia haitanensis has prominent stress-resistance characteristics and is endemic to China. Studies into the stress responses in these algae could provide valuable information on the stress-response mechanisms in the intertidal Rhodophyta. Here, the effects of salinity and light intensity on the quantum yield of photosystem II in Py. haitanensis were investigated using pulse-amplitude-modulation fluorometry. Total RNA and genomic DNA of the samples under different stress conditions were isolated. By normalizing to the genomic DNA quantity, the RNA content in each sample was evaluated. The cDNA was synthesized and the expression levels of seven potential internal control genes were evaluated using qRT-PCR method. Then, we used geNorm, a common statistical algorithm, to analyze the qRT-PCR data of seven reference genes. Potential genes that may constantly be expressed under different conditions were selected, and these genes showed stable expression levels in samples under a salinity treatment, while tubulin, glyceraldehyde-3-phosphate dehydrogenase and actin showed stability in samples stressed by strong light. Based on the results of the pulse amplitude-modulation fluorometry, an absolute quantification was performed to obtain gene copy numbers in certain stress-treated samples. The stably expressed genes as determined by the absolute quantification in certain samples conformed to the results of the geNorm screening. Based on the results of the software analysis and absolute quantification, we proposed that elongation factor 3 and 18S ribosomal RNA could be used as internal control genes when the Py. haitanensis blades were subjected to salinity stress, and that α-tubulin and 18S ribosomal RNA could be used as the internal control genes when the stress was from strong light. In general, our findings provide a convenient reference for the selection of internal control genes when designing experiments related to stress responses in Py. haitanensis.

Residues in the H+ Translocation Site Define the pKa for Sugar Binding to LacY†

Science.gov (United States)

Smirnova, Irina; Kasho, Vladimir; Sugihara, Junichi; Choe, Jun-Yong; Kaback, H. Ronald

2009-01-01

A remarkably high pKa of approximately 10.5 has been determined for sugar-binding affinity to the lactose permease of Escherichia coli (LacY), indicating that, under physiological conditions, substrate binds to fully protonated LacY. We have now systematically tested site-directed replacements for the residues involved in sugar binding, as well as H+ translocation and coupling, in order to determine which residues may be responsible for this alkaline pKa. Mutations in the sugar-binding site (Glu126, Trp151, Glu269) markedly decrease affinity for sugar but do not alter the pKa for binding. In contrast, replacements for residues involved in H+ translocation (Arg302, Tyr236, His322, Asp240, Glu325, Lys319) exhibit pKa values for sugar binding that are either shifted toward neutral pH or independent of pH. Values for the apparent dissociation constant for sugar binding (Kdapp) increase greatly for all mutants except neutral replacements for Glu325 or Lys319, which are characterized by remarkably high affinity sugar binding (i.e., low Kdapp) from pH 5.5 to pH 11. The pH dependence of the on- and off-rate constants for sugar binding measured directly by stopped-flow fluorometry implicates koff as a major factor for the affinity change at alkaline pH and confirms the effects of pH on Kdapp inferred from steady-state fluorometry. These results indicate that the high pKa for sugar binding by wild-type LacY cannot be ascribed to any single amino acid residue but appears to reside within a complex of residues involved in H+ translocation. There is structural evidence for water bound in this complex, and the water could be the site of protonation responsible for the pH dependence of sugar binding. PMID:19689129

Molecular mechanisms of Cys-loop ion channel receptor modulation by ivermectin

DEFF Research Database (Denmark)

Lynagh, Timothy; Lynch, Joseph W.

2012-01-01

Ivermectin is an anthelmintic drug that works by inhibiting neuronal activity and muscular contractility in arthropods and nematodes. It works by activating glutamate-gated chloride channels (GluClRs) at nanomolar concentrations. These receptors, found exclusively in invertebrates, belong to the ...... to the neurotransmitter binding site, thus suggesting a mechanism by which ivermectin potentiates neurotransmitter-gated currents. Together, this information provides new insights into the mechanisms of action of this important drug.......) to its site. Several lines of evidence suggest that ivermectin opens the channel pore via a structural change distinct from that induced by the neurotransmitter agonist. Conformational changes occurring at locations distant from the pore can be probed using voltage-clamp fluorometry (VCF), a technique...

Direct determination of uranium in soil, rock, ore and biological samples by laser-induced fluorometry

International Nuclear Information System (INIS)

Li Qingzhen; Zhang Yanan

1993-03-01

A laser-induced fluorometric method with modified J-22 anti-interferent fluorescent reagent for directly determining the uranium in soil, rock, ore, geochemical, biological and other samples has been studied. The effects of external ions and dilution law of sample are examined in detail. A method for correcting inner effect is proposed. A mixed solution of 0.25% NaOH-10% J-22 is prepared which can be added to the sample cuvette for direct measurement without any pre-adjustment of acidity. Therefore, it is much simpler for operation and reduces the loss and contamination of uranium. By changing the laser fluorometer sensitivity (400 ∼ 200), up to 3000 ng uranium in the cuvette can be detected. Thus, both analytical accuracy and detectable range are improved. This method is simple, rapid, accurate and applicable to various uranium-bearing samples. The detection limit is better than 0.05 μgU/g. The relative standard deviation is ≤+-5% for the rock reference samples of 0.95, 84.8, 669 and 7240 μgU/g

Studies of yeast cell oxygenation and energetics by laser fluorometry of reduced nicotinamide adenine dinucleotide

Science.gov (United States)

Pan, Fu-shih; Chen, Stephen; Mintzer, Robert A.; Chen, Chin-Tu; Schumacker, Paul

1991-03-01

It is of fundamental importance for biological scientists to assess cellular energetics. Under aerobic conditions, the tricarboxylic acid cycle (TCA cycle) is coupled with the mitochondrial electron cascade pathway to provide the cell with energy. The nicotinamide adenine dinucleotide-conjugated pair (NAD and NADH) is the coenzyme in numerous important biomedical reactions which include several important dehydrogenase reactions in the TCA cycle. Based on Le Chatelier's principle, NADH will accumulate when this energy production mechanism is impaired. The relative amounts of NAD and NADH in a cell are defined as the redox state of the cell (Williamson et.al. 1967) which provides a valuable index of cellular energetics. The sum of the amounts of NAD and NADH in a cell may be assumed to be constant during a finite time; therefore, a reliable means of measuring the NADH concentration would provide us with a useful indicator of tissue viability. Traditionally, the quantities of NADH and NAD may be measured by chemical assay methods. We can avoid these tediois analyses by exploiting the significant difference between the ultraviolet absorption spectra of this redox pair. However, because of the opacity of biological samples and the interference of other biochemicals that also absorb ultraviolet radiation, measurement of NADH and NAD+ concentrations in vivo by absorption spectroscopy is not feasible.

Hydrogeochemical and stream sediment reconnaissance of the National Uranium Resource Evaluation Program. Progress report, July--September 1976

International Nuclear Information System (INIS)

Morris, W.A.

1977-01-01

Water and/or sediment samples have been collected from some 47,000 sample locations covering about 504,000 km 2 which represents 19% of the area assigned to the LASL for the HSSR program. Slightly over half of this sampling work was done this quarter and included the first commercial, helicopter-borne sampling contract in Alaska where 4468 locations were sampled over an area of 94,000 km 2 . Thus far, uranium determinations have been made for some 12,000 water samples by fluorometry and for over 15,000 sediment samples by delayed-neutron counting. The main effort of this quarter has been directed toward completing all outstanding commercial sampling contracts and analyzing the backlog of water and sediment samples

Possible pitfalls in the search for uranium deposits using lake sediments and lake waters

International Nuclear Information System (INIS)

Levinson, A.A.; Bland, C.J.

1978-01-01

The organic-rich sediments from the centres of Canadian lakes are a valuable sampling medium in the search for uranium. However, because of the young age of hydromorphically transported uranium in these sediments, which must post-date the last period of glaciation, there has been insufficient time for the isotope 214 Bi to reach equilibrium with its ancestor 238 U. This results in equivalent uranium (eU) values significantly lower than actual uranium values determined by fluorometry or delayed neutron activation analysis. Radiometric ( 226 Ra) analyses of 12 centre-lake sediments from Seahorse Lake, Saskatchewan illustrate the potential pitfalls which may be encountered using gamma-ray spectrometry, as only 3-8% of the actual uranium present in most of the samples would have been detected. (author)

The content of minerals and trace elements in meals

International Nuclear Information System (INIS)

Bognar, A.; Schelenz, R.; Gruenewald, T.; Frahm, H.; Heine, K.; Wiechen, A.; Bundesanstalt fuer Milchforschung, Kiel

1981-07-01

Within the frame work of the research programme 'School Feeding', 68 menu items of different producers were investigated for the content of the minerals calcium, chlorine, iron, potassium, magnesium, sodium and phosphorus, and for the trace elements antimony, barium, bromine, cesium, chromium, hafnium, iridium,cobalt, copper, manganese, mercury, rubidium, scandium, selenium, silver, strontium, tin and zinc. For the analytical determination of the elements, instrumental neutron activation analysis and X-ray fluorometry were applied. The studies showed that a calculation of the content of minerals and trace elements in meals on the basis of recipes and nutritive tables for raw foods is not justified, expect for sodium and phosphorus, because incorrect results can be obtained for the majority of meals. (orig./MG) [de

Substrate and Inhibitor-Specific Conformational Changes in the Human Serotonin Transporter Revealed by Voltage-Clamp Fluorometry

DEFF Research Database (Denmark)

Söderhielm, Pella C; Andersen, Jacob; Munro, Lachlan

2015-01-01

of TM6, Ala419 in the interface between TM8 and extracellular loop (EL) 4, and Leu481 in EL5. The reporter positions were used for time-resolved measurement of conformational changes during 5-HT transport and binding of cocaine and the selective serotonin reuptake inhibitors fluoxetine and escitalopram...... changes overall, which included movements within or around TM1b, EL4, and EL5. Taken together, our data lead us to suggest that competitive inhibitors stabilize hSERT in a state that is different from the apo outward-open conformation as well as inward-facing conformations....

Fission track method for uranium ore exploration

International Nuclear Information System (INIS)

Guo Shilun; Deng Xinlu; Sun Shengfen; Meng Wu; Zhang Pengfa; Hao Xiuhong

1986-01-01

The uranium concentrations in natural water collected in the fields of uranium ore exploration with fission track method have been determined. It shows that the results of fission track method are consistent with that of fluoro-colorimetry and laser fluorometry for the same samples of water with uranium concentration in the region of 10 -4 to 10 -8 g/l. For water samples with lower uranium concentration (≤10 -8 g/l), the fission track method can still give accurate or referential results, but the other two methods failed. The reproducibility of fission track method was checked and discussed by using samples collected in the same fields of uranium ore exploration. The effects of the concentration of the impurities in natural water on determination of uranium concentration were analysed and discussed as well

Separation of thorium and uranium by liquid-liquid extraction from mixed aqueous nitric acidic-methanolic solutions

International Nuclear Information System (INIS)

Schmid, E.R.; Kenndler, E.

1976-01-01

A method is described for the separation of Th and U from each other and from other elements, usually occuring in minerals, by liquid-liquid extraction with Aliquat Nitrate (tricaprylmethyl ammoniumnitrate, 6 vol%) in benzene from a mixed solution of 2.5 M HNO 3 and methanol (1:1 volume ratio). Permissible upper concentration ratios of interfering elements, such as Li, Na, K, Mg, Ca, Al, Cu, Co(II), Fe(III), Mn(II), Ti(IV), La(III), U(VI), Cl - , ClO 4 - , SO 4 2- , PO 4 3- , have been determined. Following the separation, Th has been determined by spectrophotometry using Thorin, and U by fluorometry. Results for yield under varying conditions, together with elemental concentrations in the ppm range for U and Th in minerals, are given. (B.T.)

Detection Limits for Spectro-fluorometry: A Case Study in the Region of Finstersee, Canton Zug, Northern Switzerland

Science.gov (United States)

Otz, M. H.; Otz, H. K.; Keller, P.

2002-05-01

Synthetic fluorescent dyes, applied below the visual detection limit (< 0.1 mg/L), have been used as tracers of ground water flow paths since the beginning of the 1950s. Since 1965, we have used spectro-fluorometers with photomultipliers to measure low concentrations of fluorescent dyes in ground water in Switzerland. In collaboration with the Engineering Geology Department of the ETH, we have separated uranine at 0.1 ng/L and Na-naphtionate at 1 ng/L from background fluorescence of spring water in the Finstersee region. These values are 10-100 times lower than postulated detection limits in the literature. The use of low dye concentrations prevents a study region from being contaminated by increased background levels due to remnant dye within the aquifer, thereby leaving the region available for future dye tracing studies. Lower detection limits also can solve particular hydraulic problems where conventional methods fail and enhance the possibility for using artificial dyes in environmentally sensitive aquifer settings.

Determination of soy proteins in food samples by dispersive solid-phase immunoextraction and dynamic long-wavelength fluorometry

International Nuclear Information System (INIS)

Molina-Delgado, María Ángeles; Aguilar-Caballos, María Paz; Gomez-Hens, Agustina

2013-01-01

We report on a method for the determination of soy proteins in food samples via dispersive solid-phase immunoextraction using gold-coated magnetic nanoparticles (NPs) as a support. Soy proteins were first extracted using anti-soy protein antibodies immobilized on the NPs, and then quantified by measuring the increase in fluorescence of the long-wavelength fluorophore cresyl violet in the presence of the anionic surfactant sodium dodecyl sulfate at neutral pH in a flow system. The method involves the use of two standard or sample aliquots. The fluorescence intensity of one aliquot is directly measured whereas that of the other aliquot is measured after immunoextraction. The difference between the peak heights of both aliquots serves as the analytical information that is directly proportional to the protein concentration. The limit of detection is 0.35 mg L −1 , the linear range is from 1 to 15 mg L −1 , and the relative standard deviation is < 5 %. Proteins such as bovine serum albumin and globulins do not interfere at the same concentration level. The method was applied to the analysis of soy-based beverages and gave recoveries in the range between 80.0 and 107.3 %. (author)

Microvascular retinopathy in subjects without diabetes

DEFF Research Database (Denmark)

Munch, Inger Christine; Kessel, Line; Borch-Johnsen, Knut

2012-01-01

Purpose:  Retinal vascular lesions such as microaneurysms and haemorrhages, while typical of diabetic retinopathy, are also seen in subjects without diabetes where they are associated with elevated cardiovascular mortality. In theory, these lesions could be a consequence of past hyperglycaemia. We...... examined the prevalence and risk factors for retinopathy, including lens fluorescence, a biomarker of cumulative life-time glycaemia in adults without diabetes. Methods:  Cross-sectional population-based study of 711 subjects without diabetes (WHO 1999 criteria) aged 30-60 years, including oral glucose...... tolerance testing, clinical and laboratory examinations, non-invasive ocular lens fluorometry and seven-field fundus photography. Results:  Retinopathy was present in 8.3% (CI(95) 6.3-10.3%) of subjects. Higher systolic blood pressure (SBP) (p = 0.032), increasing body mass index (BMI) (p = 0.014) and wider...

Binding of Gallic Acid and Epigallocatechin Gallate to Heat-Unfolded Whey Proteins at Neutral pH Alters Radical Scavenging Activity of in Vitro Protein Digests.

Science.gov (United States)

Cao, Yanyun; Xiong, Youling L

2017-09-27

Preheated (80 °C for 9 min) whey protein isolate (HWPI) was reacted with 20, 120, and 240 μmol/g (protein basis) gallic acid (GA) or epigallocatechin gallate (EGCG) at neutral pH and 25 °C. Isothermal titration calorimetry and fluorometry showed a similar trend that GA binding to HWPI was moderate but weaker than EGCG binding. However, the shift of maximal fluorescence emission wavelength in opposite directions in response to GA (blue) and EGCG (red) suggests discrepant binding patterns. Electrophoresis results showed that EGCG induced formation of HWPI complexes while GA only had a marginal effect. Both free and phenolic-bound HWPI exhibited mild antiradical activity. However, when subjected to in vitro digestion, synergistic radical-scavenging activity was produced between the phenolics and peptides with the highest synergism being observed on 120 μmol/g phenolics.

Ligand-specific conformational changes in the alpha1 glycine receptor ligand-binding domain

DEFF Research Database (Denmark)

Pless, Stephan Alexander; Lynch, Joseph W

2009-01-01

, and by the antagonist, strychnine. Voltage-clamp fluorometry involves labeling introduced cysteines with environmentally sensitive fluorophores and inferring structural rearrangements from ligand-induced fluorescence changes. In the inner beta-sheet, we labeled residues in loop 2 and in binding domain loops D and E....... At each position, strychnine and glycine induced distinct maximal fluorescence responses. The pre-M1 domain responded similarly; at each of four labeled positions glycine produced a strong fluorescence signal, whereas strychnine did not. This suggests that glycine induces conformational changes...... in the inner beta-sheet and pre-M1 domain that may be important for activation, desensitization, or both. In contrast, most labeled residues in loops C and F yielded fluorescence changes identical in magnitude for glycine and strychnine. A notable exception was H201C in loop C. This labeled residue responded...

Sequential determination of arsenic, selenium, antimony, and tellurium in foods via rapid hydride evolution and atomic absorption spectrometry

International Nuclear Information System (INIS)

Fiorino, J.A.; Jones, J.W.; Capar, S.G.

1976-01-01

Analysis of acid digests of foods for As, Se, Sb, and Te was semiautomated. Hydrides generated by controlled addition of base stabilized NaBH 4 solution to acid digests are transported directly into a shielded, hydrogen (nitrogen diluted), entrained-air flame for atomic absorption spectrophotometric determination of the individual elements. The detection limits, based on 1 g of digested sample, are approximately 10 to 20 ng/g for all four elements. Measurement precision is 1 to 2 percent relative standard deviation for each element measured at 0.10 μg. A comparison is made of results of analysis of lyophilized fish tissues for As and Se by instrumental neutron activation (INAA), hydride generation with atomic absorption spectrometry, fluorometry, and spectrophotometry. NBS standard reference materials (orchard leaves and bovine liver) analyzed for As, Se, and Sb by this method show excellent agreement with certified values and with independent NAA values

Measuring the aerial application of oil dispersant from very large aircraft at moderate altitude

International Nuclear Information System (INIS)

Fay, R.R.; Giammona, C.P.; Binkley, K.; Engelhardt, F.R.

1993-01-01

As part of a study of various aspects of oil dispersant application technology, a comparison study was conducted employing the Airborne Dispersant Delivery System (ADDS) pack and the Modular Aerial Spray System (MASS) in C-130 Hercules aircraft at altitudes of 50-150 ft above ground and at speeds up to 200 knots. Dyed Corexit 9527 was applied at a target dose rate of 5 gal/acre over a collection grid composed of metal trays, Kromekote cards, oil-sensitive cards, a continuous trough, and a WRK string collector. Analysis of the collected dispersant was done by colorimetry, fluorometry, and image analysis. Correlations through the different methodologies demonstrated that high speed, moderate altitude application of oil dispersant could be successful in delivering dispersant to the surface at effective concentration and drop size. Environmental studies of the test area showed no residual dispersant in the soil following cessation of spraying. 1 ref

A database of chlorophyll a in Australian waters

Science.gov (United States)

Davies, Claire H.; Ajani, Penelope; Armbrecht, Linda; Atkins, Natalia; Baird, Mark E.; Beard, Jason; Bonham, Pru; Burford, Michele; Clementson, Lesley; Coad, Peter; Crawford, Christine; Dela-Cruz, Jocelyn; Doblin, Martina A.; Edgar, Steven; Eriksen, Ruth; Everett, Jason D.; Furnas, Miles; Harrison, Daniel P.; Hassler, Christel; Henschke, Natasha; Hoenner, Xavier; Ingleton, Tim; Jameson, Ian; Keesing, John; Leterme, Sophie C.; James McLaughlin, M.; Miller, Margaret; Moffatt, David; Moss, Andrew; Nayar, Sasi; Patten, Nicole L.; Patten, Renee; Pausina, Sarah A.; Proctor, Roger; Raes, Eric; Robb, Malcolm; Rothlisberg, Peter; Saeck, Emily A.; Scanes, Peter; Suthers, Iain M.; Swadling, Kerrie M.; Talbot, Samantha; Thompson, Peter; Thomson, Paul G.; Uribe-Palomino, Julian; van Ruth, Paul; Waite, Anya M.; Wright, Simon; Richardson, Anthony J.

2018-02-01

Chlorophyll a is the most commonly used indicator of phytoplankton biomass in the marine environment. It is relatively simple and cost effective to measure when compared to phytoplankton abundance and is thus routinely included in many surveys. Here we collate 173, 333 records of chlorophyll a collected since 1965 from Australian waters gathered from researchers on regular coastal monitoring surveys and ocean voyages into a single repository. This dataset includes the chlorophyll a values as measured from samples analysed using spectrophotometry, fluorometry and high performance liquid chromatography (HPLC). The Australian Chlorophyll a database is freely available through the Australian Ocean Data Network portal (https://portal.aodn.org.au/). These data can be used in isolation as an index of phytoplankton biomass or in combination with other data to provide insight into water quality, ecosystem state, and relationships with other trophic levels such as zooplankton or fish.

Photometric and fluorometric continuous kinetic assay of acid phosphatases with new substrates possessing longwave absorption and emission maxima.

Science.gov (United States)

Koller, E; Wolfbeis, O S

1984-11-15

A direct and continuous kinetic method for the photometric and fluorometric determination of various acid phosphatases is described. It is based on new coumarin-derived phosphates, which after enzymatic hydrolysis undergo dissociation to form intensely colored and strongly fluorescent phenolate anions. The latter have absorption maxima ranging from 385 to 505 nm, and fluorescence maxima between 470 and 595 nm. The new substrates were compared with respect to their rate of enzymatic hydrolysis, optimum pH, and detection limits of acid phosphatase from potato and wheat germ. Detection limits of 0.001 unit/ml were found by photometry, and as low as 0.00006 unit/ml by fluorometry. The principal advantages of the new substrates over existing ones are longwave absorptions and emissions, large Stokes shifts, and the low pKa values of the corresponding phenols, thus allowing a direct and continuous assay of acid phosphatase even in weakly acidic solutions.

Oil concentrations below a demulsifier treated slick

International Nuclear Information System (INIS)

Lunel, T.; Lewis, A.

1993-01-01

During field trials in the North Sea in 1992, three 20-tonne slicks of a relatively weak 30% water-in-oil emulsion were released. Two of the slicks were treated with demulsifier from spray aircraft and one of the treated slicks was sprayed with dispersant seven hours later. The experiment used flow-through fluorometry to determine oil concentrations below the control and demulsifier-treated slick. Remote sensing imagery was used to determine the area of the surface slicks. Emulsion formation was slowed down in the two demulsifier-treated slicks relative to the control slick. The demulsifier-treated slicks resulted in maximum oil concentrations in water some five times higher than the control slick and spread over a volume 10-20 times as large. The control slick was therefore more persistent on the sea surface than either of the treated slicks. 5 refs., 5 figs., 5 tabs

Hazard and risk of herbicides for marine microalgae.

Science.gov (United States)

Sjollema, Sascha B; Martínezgarcía, Gema; van der Geest, Harm G; Kraak, Michiel H S; Booij, Petra; Vethaak, A Dick; Admiraal, Wim

2014-04-01

Due to their specific effect on photosynthesis, herbicides pose a potential threat to coastal and estuarine microalgae. However, comprehensive understanding of the hazard and risk of these contaminants is currently lacking. Therefore the aim of the present study was to investigate the toxic effects of four ubiquitous herbicides (atrazine, diuron, Irgarol(®)1051 and isoproturon) and herbicide mixtures on marine microalgae. Using a Pulse Amplitude Modulation (PAM) fluorometry based bioassay we demonstrated a clear species and herbicide specific toxicity and showed that the current environmental legislation does not protect algae sufficiently against diuron and isoproturon. Although a low actual risk of herbicides in the field was demonstrated, monitoring data revealed that concentrations occasionally reach potential effect levels. Hence it cannot be excluded that herbicides contribute to observed changes in phytoplankton species composition in coastal waters, but this is likely to occur only occasionally. Copyright © 2013 Elsevier Ltd. All rights reserved.

DNA double-strand breaks measured by pulsed-field gel electrophoresis in irradiated lymphocytes from normal humans and those with Alzheimer's disease

International Nuclear Information System (INIS)

Tobi, S.E.; Itzhaki, R.F.

1993-01-01

The authors previously found that radiation-induced chromosome aberrations (dicentrics) are more numerous in lymphocytes from Alzheimer's disease (AD) patients than in those from age-matched normal individuals (Tobi et al. 1990). They have examined double-strand breaks (dsb) produced by g amma - irradiation in the DNA of AD and normal lymphocytes by using pulsed-field gel electrophoresis. The percentage of DNA migrating into the gels is an indirect measure of the number of dsb; DNA content of sequential slices of the gel was assayed by direct fluorometry and the percentage migrating was dose dependent. Results show that the level of damage is similar in AD and normal lymphocytes and preliminary assays of the rate of repair suggest that the half-time is also similar, the value being > 1 h. The latter is consistent with the known rate of rejoining of chromosome fragments in interphase lymphocytes (Pantelias and Maillie 1985). (Author)

Study of 660 nm laser-induced photoluminescence of chlorophyll-a and its applications

International Nuclear Information System (INIS)

Song, Y; Zhang, D X; Zhang, H J

2007-01-01

Based on the phenomenon of chlorophyll a photoluminescence, this paper introduces a new method to measure the chlorophyll a content, using 660nm laser diode as a new kind of light source to stimulate fluorescence as well as combining a fiber and spectrum technique. We analyze the characteristics of laser-induced fluorescence spectrum of chlorophyll a and then put forward the new method using two parameters, the relative fluorescence intensity and fluorescence intensity ratio F685/F735, to measure the chlorophyll a content in the water and green leaves respectively. The experimental results indicate that it is completely feasible to give a visual judgment for chlorophyll a content, according to the fluorescence emission spectrum of chlorophyll a. Subsequently, it is verified by three kinds of typical applications. All of these provide a new kind of light source to develop the chlorophyll a fluorometry and further give a technical foundation of on-spot monitoring the chlorophyll a content in the ocean or in green leaves

Uranium hydrogeochemical and stream sediment reconnaissance data listing release for the Three Forks Basin, Spanish Peaks, and Boulder River areas for the Bozeman NTMS quadrangle, Montana, including concentrations of forty-six additional elements

International Nuclear Information System (INIS)

Bolivar, S.L.; George, W.E.; Gallimore, D.L.; Apel, C.T.; Gansel, J.M.; Hensley, W.K.; Van Haaften, I.J.; Pirtle, J.

1980-08-01

Totals of 531 water and 1275 sediment samples were collected from 1275 stream and spring locations. Elemental concentration, field measurement, weather, geologic, and geographic data for each sample location are listed for waters and for sediments. Uranium/thorium ratios for sediment samples are also included. Water samples were initially analyzed for uranium by fluorometry. All water samples containing more than 40 ppB uranium were reanalyzed by delayed-neutron counting (DNC). All sediments were analyzed for uranium by DNC. Other elemental concentrations in sediments were determined by neutron activation analysis for 31 elements (Al, Sb, Ba, Ca, Ce, Cs, Cl, Cr, Co, Dy, Eu, Au, Hf, Fe, La, Lu, Mg, Mn, K, Rb, Sn, Sc, Na, Sr, Ta, Tb, Th, Ti, V, Yb, and Zn), by x-ray fluorescence for 13 elements (As, Bi, Cd, Cu, Pb, Mo, Ni, Nb, Se, Ag, Sn, W, and Zr), and by arc-source emission spectrography for Li and Be. Analytical results for sediments are reported as parts per million

Environmental Monitoring of Microbe Metabolic Transformation

Science.gov (United States)

Bebout, Brad (Inventor); Fleming, Erich (Inventor); Piccini, Matthew (Inventor); Beasley, Christopher (Inventor); Bebout, Leslie (Inventor)

2013-01-01

Mobile system and method for monitoring environmental parameters involved in growth or metabolic transformation of algae in a liquid. Each of one or more mobile apparati, suspended or partly or wholly submerged in the liquid, includes at least first and second environmental sensors that sense and transmit distinct first and second environmental, growth or transformation parameter values, such as liquid temperature, temperature of gas adjacent to and above the exposed surface, liquid pH, liquid salinity, liquid turbidity, O.sub.2 dissolved in the liquid, CO.sub.2 contained in the liquid, oxidization and reduction potential of the liquid, nutrient concentrations in the liquid, nitrate concentration in the liquid, ammonium concentration in the liquid, bicarbonate concentration in the liquid, phosphate concentration in the liquid, light intensity at the liquid surface, electrical conductivity of the liquid, and a parameter.alpha.(alga) associated with growth stage of the alga, using PAM fluorometry or other suitable parameter measurements.

Uranium concentrations in natural waters, South Park, Colorado

International Nuclear Information System (INIS)

Sharp, R.R. Jr.; Aamodt, P.L.

1976-08-01

During the summer of 1975, 464 water samples from 149 locations in South Park, Colorado, were taken for the Los Alamos Scientific Laboratory in order to test the field sampling and analytical methodologies proposed for the NURE Hydrogeochemical and Stream Sediment Reconnaissance for uranium in the Rocky Mountain states and Alaska. The study showed, in the South Park area, that the analytical results do not vary significantly between samples which were untreated, filtered and acidified, filtered only, or acidified only. Furthermore, the analytical methods of fluorometry and delayed-neutron counting, as developed at the LASL for the reconnaissance work, provide fast, adequately precise, and complementary procedures for analyzing a broad range of uranium in natural waters. The data generated using this methodology does appear to identify uraniferous areas, and when applied using sound geochemical, geological, and hydrological principles, should prove a valuable tool in reconnaissance surveying to delineate new districts or areas of interest for uranium exploration

Comparative assessment of single and joint effects of diuron and Irgarol 1051 on Arctic and temperate microalgae using chlorophyll a fluorescence imaging

KAUST Repository

Kottuparambil, Sreejith

2017-02-06

Ship groundings and ice-breakers can cause pollution of the polar environment with antifouling biocides such as diuron and Irgarol 1051. The present study used pulse amplitude modulated fluorometry to compare single and joint toxicities of diuron and Irgarol 1051 on two freshwater taxa of microalgae (Chlorella and Chlamydomonas) originating from Arctic and temperate regions. 30min acute toxicity tests using chlorophyll a (Chl a) fluorescence revealed that Arctic strains of microalgae were more sensitive to herbicides than their temperate counterparts. Diuron and Irgarol 1051 had equal toxicities in the Arctic species, while Irgarol 1051 was more toxic (EC50=5.55–14.70μgL−1) than diuron (EC50=12.90–>40μgL−1) in the temperate species. Toxicity assessment of various mixtures of diuron and Irgarol 1051 revealed antagonistic, additive, and synergistic effects. Our data suggest that herbicides can adversely affect photosynthesis in Arctic microalgae at relatively low levels, and their impact can increase under complex mixture conditions.

Allosteric substrate switching in a voltage-sensing lipid phosphatase.

Science.gov (United States)

Grimm, Sasha S; Isacoff, Ehud Y

2016-04-01

Allostery provides a critical control over enzyme activity, biasing the catalytic site between inactive and active states. We found that the Ciona intestinalis voltage-sensing phosphatase (Ci-VSP), which modifies phosphoinositide signaling lipids (PIPs), has not one but two sequential active states with distinct substrate specificities, whose occupancy is allosterically controlled by sequential conformations of the voltage-sensing domain (VSD). Using fast fluorescence resonance energy transfer (FRET) reporters of PIPs to monitor enzyme activity and voltage-clamp fluorometry to monitor conformational changes in the VSD, we found that Ci-VSP switches from inactive to a PIP3-preferring active state when the VSD undergoes an initial voltage-sensing motion and then into a second PIP2-preferring active state when the VSD activates fully. This two-step allosteric control over a dual-specificity enzyme enables voltage to shape PIP concentrations in time, and provides a mechanism for the complex modulation of PIP-regulated ion channels, transporters, cell motility, endocytosis and exocytosis.

Dimerization of the voltage-sensing phosphatase controls its voltage-sensing and catalytic activity.

Science.gov (United States)

Rayaprolu, Vamseedhar; Royal, Perrine; Stengel, Karen; Sandoz, Guillaume; Kohout, Susy C

2018-05-07

Multimerization is a key characteristic of most voltage-sensing proteins. The main exception was thought to be the Ciona intestinalis voltage-sensing phosphatase (Ci-VSP). In this study, we show that multimerization is also critical for Ci-VSP function. Using coimmunoprecipitation and single-molecule pull-down, we find that Ci-VSP stoichiometry is flexible. It exists as both monomers and dimers, with dimers favored at higher concentrations. We show strong dimerization via the voltage-sensing domain (VSD) and weak dimerization via the phosphatase domain. Using voltage-clamp fluorometry, we also find that VSDs cooperate to lower the voltage dependence of activation, thus favoring the activation of Ci-VSP. Finally, using activity assays, we find that dimerization alters Ci-VSP substrate specificity such that only dimeric Ci-VSP is able to dephosphorylate the 3-phosphate from PI(3,4,5)P 3 or PI(3,4)P 2 Our results indicate that dimerization plays a significant role in Ci-VSP function. © 2018 Rayaprolu et al.

Allosteric substrate switching in a voltage sensing lipid phosphatase

Science.gov (United States)

Grimm, Sasha S.; Isacoff, Ehud Y.

2016-01-01

Allostery provides a critical control over enzyme activity, biasing the catalytic site between inactive and active states. We find the Ciona intestinalis voltage-sensing phosphatase (Ci-VSP), which modifies phosphoinositide signaling lipids (PIPs), to have not one but two sequential active states with distinct substrate specificities, whose occupancy is allosterically controlled by sequential conformations of the voltage sensing domain (VSD). Using fast FRET reporters of PIPs to monitor enzyme activity and voltage clamp fluorometry to monitor conformational changes in the VSD, we find that Ci-VSP switches from inactive to a PIP3-preferring active state when the VSD undergoes an initial voltage sensing motion and then into a second PIP2-preferring active state when the VSD activates fully. This novel 2-step allosteric control over a dual specificity enzyme enables voltage to shape PIP concentrations in time, and provides a mechanism for the complex modulation of PIP-regulated ion channels, transporters, cell motility and endo/exocytosis. PMID:26878552

Uranium hydrogeochemical and stream sediment reconnaissance of the Bozeman NTMS quadrangle, Montana, including concentrations of forty-two additional elements

International Nuclear Information System (INIS)

Bolivar, S.L.; Hensley, W.K.; Van Haaften, I.J.; Pirtle, J.; George, W.E.; Gallimore, D.; Apel, C.; Hansel, J.

1980-07-01

This report contains uranium analyses for 1251 water samples and multielement analyses for 1536 sediment samples. Sediments were analyzed for uranium and thorium as well as aluminum, antimony, barium, beryllium, bismuth, cadmium, calcium, cerium, cesium, chlorine, chromium, cobalt, copper, dysprosium, europium, gold, hafnium, iron, lanthanum, lead, lithium, lutetium, magnesium, manganese, nickel, niobium, potassium, rubidium, samarium, scandium, silver, sodium, strontium, tantalum, terbium, tin, titanium, tungsten, vanadium, ytterbium, and zinc. Water samples were initially analyzed for uranium by fluorometry. All water samples containing more than 40 ppB uranium were reanalyzed by delayed-neutron counting (DNC). All sediments were analyzed for uranium by DNC. Other elemental concentrations in sediments were determined by neutron activation analysis for 31 elements, by x-ray fluorescence for 9 elements, and by arc-source emission spectrography for 2 elements. Analytical results for sediments are reported as parts per million. Descriptions of procedures used for analysis of water and sediment samples as well as analytical precisions and detection limits are given

Ecologically acceptable usage of derivatives of essential oil of sweet basil, Ocimum basilicum, as antifeedants against larvae of the gypsy moth, Lymantria dispar.

Science.gov (United States)

Popović, Zorica; Kostić, Miroslav; Stanković, Sladjan; Milanović, Slobodan; Sivčev, Ivan; Kostić, Igor; Kljajić, Petar

2013-01-01

Abstract Ethanol solutions of five fractions obtained from essential oil of sweet basil Ocimum basilicum L. (Lamiales: Lamiaceae) (F1-F5) were tested for their antifeedant properties against 2(nd) instar gypsy moth larvae, Lymantria dispar L. (Lepidoptera: Lymantriidae), in laboratory non-choice and feeding-choice experiments. Prior to bioassays, the chemical composition of each fraction was determined by gas chromatography analyses. Significant larval deterrence from feeding was achieved by application of tested solutions to fresh leaves of the host plant. The most effective were were F1 (0.5%), F4 (0.05, 0.1, and 0.5%), and F5 (0.1 and 0.5%), which provided an antifeedant index > 80% after five days. A low rate of larval mortality was observed in no-choice bioassay. In situ screening of chlorophyll fluorescence as an indicator of plant stress level (assessed by the induced fluorometry) confirmed that the tested compounds did not cause alternations in the photosynthetic efficiency of treated leaves.

Photosynthetic response of two seaweed species along an urban pollution gradient: evidence of selection of pollution-tolerant species.

Science.gov (United States)

Scherner, F; Bonomi Barufi, J; Horta, P A

2012-11-01

Urbanization leads to the expansion of ephemeral seaweed species and the decline of important perennial, canopy-forming seaweed species. Understanding the mechanisms that lead to these changes is a current challenge. In the present study, laboratory assays and field transplantations were performed with two seaweed species: the perennial, canopy-forming seaweed Sargassum stenophyllum and the ephemeral seaweed Ulva lactuca. Photosynthetic efficiency was assessed using modulated chlorophyll fluorometry. Brief exposure to urban waters does not appear to be a major stressor to the photosynthetic efficiency of either species. However, after 26 days of transplantation in urban waters, S. stenophyllum declined, whereas U. lactuca had enhanced photosynthetic efficiency. This difference reflects their divergent abilities to regulate the energy distribution at the PSII and shows that urban stressors alter these mechanisms. Our results provide evidence of the physiological causes for the decline of Sargassum species and the expansion of Ulva species in impacted urban areas. Copyright © 2012 Elsevier Ltd. All rights reserved.

Comparative assessment of single and joint effects of diuron and Irgarol 1051 on Arctic and temperate microalgae using chlorophyll a fluorescence imaging

KAUST Repository

Kottuparambil, Sreejith; Brown, Murray T.; Park, Jihae; Choi, Soyeon; Lee, Hojun; Choi, Han-Gu; Depuydt, Stephen; Han, Taejun

2017-01-01

Ship groundings and ice-breakers can cause pollution of the polar environment with antifouling biocides such as diuron and Irgarol 1051. The present study used pulse amplitude modulated fluorometry to compare single and joint toxicities of diuron and Irgarol 1051 on two freshwater taxa of microalgae (Chlorella and Chlamydomonas) originating from Arctic and temperate regions. 30min acute toxicity tests using chlorophyll a (Chl a) fluorescence revealed that Arctic strains of microalgae were more sensitive to herbicides than their temperate counterparts. Diuron and Irgarol 1051 had equal toxicities in the Arctic species, while Irgarol 1051 was more toxic (EC50=5.55–14.70μgL−1) than diuron (EC50=12.90–>40μgL−1) in the temperate species. Toxicity assessment of various mixtures of diuron and Irgarol 1051 revealed antagonistic, additive, and synergistic effects. Our data suggest that herbicides can adversely affect photosynthesis in Arctic microalgae at relatively low levels, and their impact can increase under complex mixture conditions.

Uranium concentrations in natural waters, South Park, Colorado. [Part of National Uranium Resource Evaluation program

Energy Technology Data Exchange (ETDEWEB)

Sharp, R.R. Jr.; Aamodt, P.L.

1976-08-01

During the summer of 1975, 464 water samples from 149 locations in South Park, Colorado, were taken for the Los Alamos Scientific Laboratory in order to test the field sampling and analytical methodologies proposed for the NURE Hydrogeochemical and Stream Sediment Reconnaissance for uranium in the Rocky Mountain states and Alaska. The study showed, in the South Park area, that the analytical results do not vary significantly between samples which were untreated, filtered and acidified, filtered only, or acidified only. Furthermore, the analytical methods of fluorometry and delayed-neutron counting, as developed at the LASL for the reconnaissance work, provide fast, adequately precise, and complementary procedures for analyzing a broad range of uranium in natural waters. The data generated using this methodology does appear to identify uraniferous areas, and when applied using sound geochemical, geological, and hydrological principles, should prove a valuable tool in reconnaissance surveying to delineate new districts or areas of interest for uranium exploration.

Succession and physiological health of freshwater microalgal fouling in a Tasmanian hydropower canal.

Science.gov (United States)

Perkins, Kathryn J; Andrewartha, Jessica M; McMinn, Andrew; Cook, Suellen S; Hallegraeff, Gustaaf M

2010-08-01

Freshwater microalgal biofouling in hydropower canals in Tarraleah, Tasmania, is dominated by a single diatom species, Gomphonema tarraleahae. The microfouling community is under investigation with the aim of reducing its impact on electricity generation. Species succession was investigated using removable glass slides. Fouled slides were examined microscopically and for chlorophyll a biomass. Chl a biomass increased steeply after 8 weeks (0.09-0.87 mg m(-2)), but increased much earlier on slides surrounded by a biofouled inoculum. Succession began with low profile diatoms such as Tabellaria flocculosa, progressing to stalked diatoms such as Gomphonema spp. and Cymbella aspera. Few chlorophytes and no filamentous algae were present. Pulse amplitude modulated fluorometry was used to measure the physiological health of fouling on the canal wall. Maximum quantum yield (F(v)/F(m)) measurements were consistently <0.18, indicating that the fouling mat consisted of dead or dying algae. The succession and physiological health of cells in the fouling community has broad implications for mitigation techniques used.

Application of ultraviolet fluorometry and excitation-emission matrix spectroscopy (EEMS) to fingerprint oil and chemically dispersed oil in seawater.

Science.gov (United States)

Bugden, J B C; Yeung, C W; Kepkay, P E; Lee, K

2008-04-01

Excitation-emission matrix spectroscopy (EEMS) was used to characterize the ultra violet fluorescence fingerprints of eight crude oils (with a 14,470-fold range of dynamic viscosity) in seawater. When the chemical dispersant Corexit 9500 was mixed with the oils prior to their dispersion in seawater, the fingerprints of each oil changed primarily as an increase in fluorescence over an emission band centered on 445 nm. In order to simplify the wealth of information available in the excitation-emission matrix spectra (EEMs), two ratios were calculated. A 66-90% decrease in the slope ratio was observed with the addition of Corexit. When the slope ratios were reduced in complexity to intensity ratios, similar trends were apparent. As a result either of the ratios could be used as a simple and rapid means of identifying and monitoring chemically dispersed oil in the open ocean.

Determination of the Critical Micelle Concentration of Neutral and Ionic Surfactants with Fluorometry, Conductometry, and Surface Tension-A Method Comparison.

Science.gov (United States)

Scholz, Norman; Behnke, Thomas; Resch-Genger, Ute

2018-01-01

Micelles are of increasing importance as versatile carriers for hydrophobic substances and nanoprobes for a wide range of pharmaceutical, diagnostic, medical, and therapeutic applications. A key parameter indicating the formation and stability of micelles is the critical micelle concentration (CMC). In this respect, we determined the CMC of common anionic, cationic, and non-ionic surfactants fluorometrically using different fluorescent probes and fluorescence parameters for signal detection and compared the results with conductometric and surface tension measurements. Based upon these results, requirements, advantages, and pitfalls of each method are discussed. Our study underlines the versatility of fluorometric methods that do not impose specific requirements on surfactants and are especially suited for the quantification of very low CMC values. Conductivity and surface tension measurements yield smaller uncertainties particularly for high CMC values, yet are more time- and substance consuming and not suitable for every surfactant.

Distribution and development of algal photosynthesis in sea ice during a spring bloom: A novel application of Imaging-PAM fluorometry

DEFF Research Database (Denmark)

Hawes, Ian; Lund-Hansen, Lars Chresten; Sorrell, Brian Keith

were able to migrate with ice growth. Furthermore, increases in maximum fluorescence (Fm) and photosynthesis yield (Y) allowed us to document the progression of a spring bloom in algal biomass and activity. Maximum light intensities in the ice algal communities were ca. 40 µmol PPFD m-2 s-1 and light......Sea ice algae are important marine primary producers that may contribute as much as 15% of total global carbon fixation. The difficulties of studying photosynthesis in ice have, however, hampered accurate assessments of their productivity, and much previous research has involved liquid...... cores from the Kangerlussuaq Fjord, Greenland, over two weeks in March 2011, and used Imaging-PAM to explore the spatial distribution of algal biomass (predominantly pennate diatoms), fluorescence yield, and light response curves. Algae were largely confined to the lowest 1 cm of the ice column, where...

Extraction of uranium from sea water with the granular composite adsorbent by using the fixed bed

International Nuclear Information System (INIS)

Katoh, Shunsaku; Sakane, Kohji; Hirotsu, Takahiro; Fujii, Ayako; Kitamura, Takao

1981-01-01

To clarify the technical problems existing in the extraction process of uranium from sea water, uranium was extracted from natural sea water, with the granulated C-Ti-OH composite adsorbent. The adsorption of uranium from sea water was carried out by using the fixed bed that had been designed in our laboratory. The uranium recovery from the sea water was 13.9% in the adsorption process of 56 d. The adsorbed uranium was eluted from the adsorbent with 0.5 N NaHCO 3 -0.5N Na 2 CO 3 soln. at 70 0 C. The elution recovery was 97.4% for 35 h. The uranium contained in the eluate was concentrated twenty times as much as in the anion exchange process, and then 100 times in the solvent extraction process with oxine-chloroform and TOA-kerosene. About 0.7 g of yellow cake was prepared from natural sea water, and it was identified to be pure 2UO 2 .NH 3 .3H 2 O by X-ray diffraction method and X-ray fluorometry. (author)

Interactions in the aqueous phase and adsorption at the air-water interface of caseinoglycomacropeptide (GMP) and beta-lactoglobulin mixed systems.

Science.gov (United States)

Martinez, María J; Sánchez, Cecilio Carrera; Patino, Juan M Rodríguez; Pilosof, Ana M R

2009-01-01

The aim of this work was to study the interactions and adsorption of caseinoglycomacropeptide (GMP) and GMP:beta-lactoglobulin (beta-lg) mixed system in the aqueous phase and at the air-water interface. The existence of associative interactions between GMP and beta-lg in the aqueous phase was investigated by dynamic light scattering, differential scanning calorimetry (DSC), fluorometry and native PAGE-electrophoresis. The surface pressure isotherm and the static and dynamic surface pressure were determined by tensiometry and surface dilatational properties. The results showed that GMP presented higher surface activity than beta-lg at a concentration of 4%wt but beta-lg showed higher film forming ability. In the mixed systems beta-lg dominated the static and dynamic surface pressure and the rheological properties of interfacial films suggesting that beta-lg hinders GMP adsorption because, in simple competition, GMP should dominate because of its higher surface activity. The surface predominance of beta-lg can be attributed to binding of GMP to beta-lg in the aqueous phase that prevents GMP adsorption on its own.

Modification of radiation sensitivity by edaravone

International Nuclear Information System (INIS)

Sasano, Nakashi; Enomoto, Atsushi; Miyagawa, Kiyoshi; Hosoi, Yoshio; Nakagawa, Keiichi

2009-01-01

Studied was the effect of edaravone (E), a clinical therapeutic drug for brain infarction possessing properties of free radical scavenger, on apoptosis in vitro. Human T-cell leukemic MOLT-4 cells and p53-knockdown MOLT-4 cells which overexpressing short hairpin type p53 by siRNA treatment were X-irradiated by Shimadzu Pantak HF 350 machine at 135-140 cGy/min for 2-5 Gy in total, in the presence of E. Cell death was estimated by dye-exclusion, apoptosis by EPICS flow cytometry, cellular reactive oxygen species by chloromethyl-2',7'-dichlorodihydrofluoresein diacetate (CM-H 2 -DCFDA) fluorometry, and p53-related protein expression by Western blotting. E was found to have a radio-protective effect on cells at high concentrations (e.g., 2.7 and 3 mg/mL, 2Gy) and radio-sensitizing action at low concentrations (e.g., 0.15-1.5 mg/mL). Clinical plasma levels of E have been reportedly far lower than the concentrations in this experiment: ideally, E can sensitize tumor cells to radiation and protect normal cells from. (K.T.)

Conformational dynamics and role of the acidic pocket in ASIC pH-dependent gating.

Science.gov (United States)

Vullo, Sabrina; Bonifacio, Gaetano; Roy, Sophie; Johner, Niklaus; Bernèche, Simon; Kellenberger, Stephan

2017-04-04

Acid-sensing ion channels (ASICs) are proton-activated Na + channels expressed in the nervous system, where they are involved in learning, fear behaviors, neurodegeneration, and pain sensation. In this work, we study the role in pH sensing of two regions of the ectodomain enriched in acidic residues: the acidic pocket, which faces the outside of the protein and is the binding site of several animal toxins, and the palm, a central channel domain. Using voltage clamp fluorometry, we find that the acidic pocket undergoes conformational changes during both activation and desensitization. Concurrently, we find that, although proton sensing in the acidic pocket is not required for channel function, it does contribute to both activation and desensitization. Furthermore, protonation-mimicking mutations of acidic residues in the palm induce a dramatic acceleration of desensitization followed by the appearance of a sustained current. In summary, this work describes the roles of potential pH sensors in two extracellular domains, and it proposes a model of acidification-induced conformational changes occurring in the acidic pocket of ASIC1a.

Metal speciation in surface waters of the Great Lakes region. Annual report (final)

International Nuclear Information System (INIS)

Giesy, J.P.

1983-01-01

Predicting the dynamics of trace metals such as uranium by thermodynamic prediction models is contingent upon accurate stability constants. Conditional stability constants (K') for the binding of uranyl ion UO2(+2) to humic substances were determined by both Scatchard analyses and by fitting the data to a Gaussian model of multiple sites. UO2(+2) was separated from that which was bound to humic material by ion exchange. Uranium concentration was measured by laser fluorometry. There is a broad range of strengths of sites such that K' was dependent on the UO2(+2)/humic ratio. The maximum binding capacity of the humic material was .000483 M/g. The log K' determined for log metal/ligand ratios of -1.5 to -0.5, the ecologically significant range of interest for UO2(+2), which was from 10 - 50 microgram/l at the carbon concentration of 2.36 mg/l used in this study, was determined by Scatchard analysis to be 7.38. The estimate based on a Gaussian distribution was 6.85

Ecotoxicology of bromoacetic acid on estuarine phytoplankton.

Science.gov (United States)

Gordon, Ana R; Richardson, Tammi L; Pinckney, James L

2015-11-01

Bromoacetic acid is formed when effluent containing chlorine residuals react with humics in natural waters containing bromide. The objective of this research was to quantify the effects of bromoacetic acid on estuarine phytoplankton as a proxy for ecosystem productivity. Bioassays were used to measure the EC50 for growth in cultured species and natural marine communities. Growth inhibition was estimated by changes in chlorophyll a concentrations measured by fluorometry and HPLC. The EC50s for cultured Thalassiosira pseudonana were 194 mg L(-1), 240 mg L(-1) for Dunaliella tertiolecta and 209 mg L(-1) for Rhodomonas salina. Natural phytoplankton communities were more sensitive to contamination with an EC50 of 80 mg L(-1). Discriminant analysis suggested that bromoacetic acid additions cause an alteration of phytoplankton community structure with implications for higher trophic levels. A two-fold EC50 decrease in mixed natural phytoplankton populations affirms the importance of field confirmation for establishing water quality criteria. Copyright © 2015 Elsevier Ltd. All rights reserved.

Recommendations for fluorescence instrument qualification: the new ASTM Standard Guide.

Science.gov (United States)

DeRose, Paul C; Resch-Genger, Ute

2010-03-01

Aimed at improving quality assurance and quantitation for modern fluorescence techniques, ASTM International (ASTM) is about to release a Standard Guide for Fluorescence, reviewed here. The guide's main focus is on steady state fluorometry, for which available standards and instrument characterization procedures are discussed along with their purpose, suitability, and general instructions for use. These include the most relevant instrument properties needing qualification, such as linearity and spectral responsivity of the detection system, spectral irradiance reaching the sample, wavelength accuracy, sensitivity or limit of detection for an analyte, and day-to-day performance verification. With proper consideration of method-inherent requirements and limitations, many of these procedures and standards can be adapted to other fluorescence techniques. In addition, procedures for the determination of other relevant fluorometric quantities including fluorescence quantum yields and fluorescence lifetimes are briefly introduced. The guide is a clear and concise reference geared for users of fluorescence instrumentation at all levels of experience and is intended to aid in the ongoing standardization of fluorescence measurements.

Seasonal variability in irradiance affects herbicide toxicity to the marine flagellate Dunaliella tertiolecta

Directory of Open Access Journals (Sweden)

Sascha eSjollema

2014-06-01

Full Text Available Photosynthetically Active Radiation (PAR and Ultraviolet Radiation (UVR of the solar spectrum affect microalgae directly and modify the toxicity of phytotoxic compounds present in water. As a consequence seasonal variable PAR and UVR levels are likely to modulate the toxic pressure of contaminants in the field. Therefore the present study aimed to determine the toxicity of two model contaminants, the herbicides diuron and Irgarol®1051, under simulated irradiance conditions mimicking different seasons. Irradiance conditions of spring and autumn were simulated with a set of Light Emitting Diodes (LEDs. Toxicity of both herbicides was measured individually and in a mixture by determining the inhibition of photosystem II efficiency (ΦPSII of the marine flagellate Dunaliella teriolecta using Pulse Amplitude Modulation (PAM fluorometry. Toxicity of the single herbicides was higher under simulated spring irradiance than under autumn irradiance and this effect was also observed for mixtures of the herbicides. This irradiance dependent toxicity indicates that herbicide toxicity in the field is seasonally variable. Consequently toxicity tests under standard light conditions may overestimate or underestimate the toxic effect of phytotoxic compounds.

A new mechanistic understanding of light-limitation in the seagrass Zostera muelleri.

Science.gov (United States)

Davey, Peter A; Pernice, Mathieu; Ashworth, Justin; Kuzhiumparambil, Unnikrishnan; Szabó, Milán; Dolferus, Rudy; Ralph, Peter J

2018-03-01

In this study we investigated the effect of light-limitation (∼20 μmol photons m -2  s -1 ) on the southern hemisphere seagrass, Zostera muelleri. RNA sequencing, chlorophyll fluorometry and HPLC techniques were used to investigate how the leaf-specific transcriptome drives changes in photosynthesis and photo-pigments in Z. muelleri over 6 days. 1593 (7.51%) genes were differentially expressed on day 2 and 1481 (6.98%) genes were differentially expressed on day 6 of the experiment. Differential gene expression correlated with significant decreases in rETR Max , I k , an increase in Yi (initial photosynthetic quantum yield of photosystem II), and significant changes in pigment composition. Regulation of carbohydrate metabolism was observed along with evidence that abscisic acid may serve a role in the low-light response of this seagrass. This study provides a novel understanding of how Z. muelleri responds to light-limitation in the marine water column and provides potential molecular markers for future conservation monitoring efforts. Copyright © 2018 Elsevier Ltd. All rights reserved.

Photosynthetic efficiency of Pedunculate oak seedlings under simulated water stress

Directory of Open Access Journals (Sweden)

Popović Zorica

2010-01-01

Full Text Available Photosynthetic performance of seedlings of Quercus robur exposed to short-term water stress in the laboratory conditions was assessed through the method of induced fluorometry. The substrate for seedlings was clayey loam, with the dominant texture fraction made of silt, followed by clay and fine sand, with total porosity 68.2%. Seedlings were separated in two groups: control (C (soil water regime in pots was maintained at the level of field water capacity and treated (water-stressed, WS (soil water regime was maintained in the range of wilting point and lentocapillary capacity. The photosynthetic efficiency was 0.642±0.25 and 0.522±0.024 (WS and C, respectively, which was mostly due to transplantation disturbances and sporadic leaf chlorosis. During the experiment Fv/Fm decreased in both groups (0.551±0.0100 and 0.427±0.018 in C and WS, respectively. Our results showed significant differences between stressed and control group, in regard to both observed parameters (Fv/Fm and T½. Photosynthetic efficiency of pedunculate oak seedlings was significantly affected by short-term water stress, but to a lesser extent than by sufficient watering.

Changes in parasympathetic system in medulla oblongata in male pigs in the course of tachycardia-induced cardiomyopathy.

Science.gov (United States)

Tomaszek, Alicja; Kiczak, Liliana; Bania, Jacek; Krupa, Paweł; Pasławska, Urszula; Zacharski, Maciej; Janiszewski, Adrian; Stefaniak, Tadeusz; Zyśko, Dorota; Ardehali, Hossein; Jankowska, Ewa A; Ponikowski, Piotr

2013-10-01

Autonomic imbalance constituting a fundamental feature of heart failure (HF) has been assessed mainly at the periphery. Changes in the functioning of autonomic centers in the brain remain unclear. We investigated the molecular elements of parasympathetic system, i.e. α7 nicotinic acetylcholine receptor (α7nAChR) and enzymes metabolizing acetylcholine (acetylcholinesterase, AChE, choline acetyltransferase, ChAT) in medulla oblongata (MO) of male pigs with chronic tachycardia-induced cardiomyopathy. The mRNA levels of AChE, ChAT, α7nAChR and X-box binding protein 1 (spliced form, XBP1s) in MO were analyzed using qPCR, AChE and ChAT activities using spectrophotometry, proteasome activity using fluorometry, and the protein level of α7nAChR using Western blotting. The development of systolic HF was accompanied by an increase in circulating catecholamines, a decrease in the AChE and α7nAChR mRNA in MO, an increase in AChE activity (all pmedulla oblongata during the progression of systolic non-ischemic heart failure in male pigs, indicating a functional link between MO and heart in HF. Copyright © 2013 Elsevier B.V. All rights reserved.

Modification of radiation sensitivity by edaravone

Energy Technology Data Exchange (ETDEWEB)

Sasano, Nakashi; Enomoto, Atsushi; Miyagawa, Kiyoshi [Tokyo Univ., Graduate School of Medicine, Tokyo (Japan); Hosoi, Yoshio [Niigata Univ., School of Health Sciences, Niigata, Niigata (Japan); Nakagawa, Keiichi [Tokyo Univ., Hospital, Tokyo (Japan)

2009-03-15

Studied was the effect of edaravone (E), a clinical therapeutic drug for brain infarction possessing properties of free radical scavenger, on apoptosis in vitro. Human T-cell leukemic MOLT-4 cells and p53-knockdown MOLT-4 cells which overexpressing short hairpin type p53 by siRNA treatment were X-irradiated by Shimadzu Pantak HF 350 machine at 135-140 cGy/min for 2-5 Gy in total, in the presence of E. Cell death was estimated by dye-exclusion, apoptosis by EPICS flow cytometry, cellular reactive oxygen species by chloromethyl-2',7'-dichlorodihydrofluoresein diacetate (CM-H{sub 2}-DCFDA) fluorometry, and p53-related protein expression by Western blotting. E was found to have a radio-protective effect on cells at high concentrations (e.g., 2.7 and 3 mg/mL, 2Gy) and radio-sensitizing action at low concentrations (e.g., 0.15-1.5 mg/mL). Clinical plasma levels of E have been reportedly far lower than the concentrations in this experiment: ideally, E can sensitize tumor cells to radiation and protect normal cells from. (K.T.)

Activation analysis of selenium in odorous vegetable foods

International Nuclear Information System (INIS)

Suzuki, Shogo; Hirai, Shoji; Noda, Katsuhiko.

1981-01-01

The selenium in odorous vegetables was analyzed by nondestructive neutron activation analysis using 75 Se, by the γ-ray coincidence method with a Ge(Li) and a NaI(Tl) detectors of definite energy ranges. By means of the coincidence counting, the background spectrum in 75 Se in the vicinity of 265 KeV was able to be reduced to about 1/20 of that by the former detector alone, so that the 75 Se detection sensitivity was raised over fourfold. Thus the selenium in odorous vegetables was able to be determined down to the content as low as 0.02 μg/g. The selenium content in garlic bulbs was 0.02 - 0.31 μg/g, and in onion bulbs 0.02 - 0.05 μg/g, both of which agreed well with those by fluorometry. In other odorous vegetables, the selenium content was as little as 0.1 μg/g or lower. It has been said that the selenium content is relatively large along with sulfur because of the same group, but it was found to be fairly small in the odorous vegetables. (Mori, K.)

Recent developments in cyanide detection: A review

Energy Technology Data Exchange (ETDEWEB)

Ma Jian [Department of Chemistry and Biochemistry, University of Texas, 700 Planetarium Place, Arlington, TX 76019-0065 (United States); Dasgupta, Purnendu K., E-mail: Dasgupta@uta.edu [Department of Chemistry and Biochemistry, University of Texas, 700 Planetarium Place, Arlington, TX 76019-0065 (United States)

2010-07-19

The extreme toxicity of cyanide and environmental concerns from its continued industrial use continue to generate interest in facile and sensitive methods for cyanide detection. In recent years, there is also additional recognition of HCN toxicity from smoke inhalation and potential use of cyanide as a weapon of terrorism. This review summarizes the literature since 2005 on cyanide measurement in different matrices ranging from drinking water and wastewater, to cigarette smoke and exhaled breath to biological fluids like blood, urine and saliva. The dramatic increase in the number of publications on cyanide measurement is indicative of the great interest in this field not only from analytical chemists, but also researchers from diverse environmental, medical, forensic and clinical arena. The recent methods cover both established and emerging analytical disciplines and include naked eye visual detection, spectrophotometry/colorimetry, capillary electrophoresis with optical absorbance detection, fluorometry, chemiluminescence, near-infrared cavity ring down spectroscopy, atomic absorption spectrometry, electrochemical methods (potentiometry/amperometry/ion chromatography-pulsed amperometry), mass spectrometry (selected ion flow tube mass spectrometry, electrospray ionization mass spectrometry, gas chromatography-mass spectrometry), gas chromatography (nitrogen phosphorus detector, electron capture detector) and quartz crystal mass monitors.

Recent developments in cyanide detection: A review

International Nuclear Information System (INIS)

Ma Jian; Dasgupta, Purnendu K.

2010-01-01

The extreme toxicity of cyanide and environmental concerns from its continued industrial use continue to generate interest in facile and sensitive methods for cyanide detection. In recent years, there is also additional recognition of HCN toxicity from smoke inhalation and potential use of cyanide as a weapon of terrorism. This review summarizes the literature since 2005 on cyanide measurement in different matrices ranging from drinking water and wastewater, to cigarette smoke and exhaled breath to biological fluids like blood, urine and saliva. The dramatic increase in the number of publications on cyanide measurement is indicative of the great interest in this field not only from analytical chemists, but also researchers from diverse environmental, medical, forensic and clinical arena. The recent methods cover both established and emerging analytical disciplines and include naked eye visual detection, spectrophotometry/colorimetry, capillary electrophoresis with optical absorbance detection, fluorometry, chemiluminescence, near-infrared cavity ring down spectroscopy, atomic absorption spectrometry, electrochemical methods (potentiometry/amperometry/ion chromatography-pulsed amperometry), mass spectrometry (selected ion flow tube mass spectrometry, electrospray ionization mass spectrometry, gas chromatography-mass spectrometry), gas chromatography (nitrogen phosphorus detector, electron capture detector) and quartz crystal mass monitors.

Diel tuning of photosynthetic systems in ice algae at Saroma-ko Lagoon, Hokkaido, Japan

Science.gov (United States)

Aikawa, Shimpei; Hattori, Hiroshi; Gomi, Yasushi; Watanabe, Kentaro; Kudoh, Sakae; Kashino, Yasuhiro; Satoh, Kazuhiko

Ice algae are the major primary producers in seasonally ice-covered oceans during the cold season. Diurnal change in solar radiation is inevitable for ice algae, even beneath seasonal sea ice in lower-latitude regions. In this work, we focused on the photosynthetic response of ice algae under diurnally changing irradiance in Saroma-ko Lagoon, Japan. Photosynthetic properties were assessed by pulse-amplitude modulation (PAM) fluorometry. The species composition remained almost the same throughout the investigation. The maximum electron transport rate ( rETRmax), which indicates the capacity of photosynthetic electron transport, increased from sunrise until around noon and decreased toward sunset, with no sign of the afternoon depression commonly observed in other photosynthetic organisms. The level of non-photochemical quenching, which indicates photoprotection activity by dissipating excess light energy via thermal processes, changed with diurnal variations in irradiance. The pigment composition appeared constant, except for xanthophyll cycle pigments, which changed irrespective of irradiance. These results indicate that ice algae tune their photosynthetic system harmonically to achieve efficient photosynthesis under diurnally changing irradiance, while avoiding damage to photosystems. This regulation system may be essential for productive photosynthesis in ice algae.

Nanoscale isoindigo-carriers: self-assembly and tunable properties

Directory of Open Access Journals (Sweden)

Tatiana N. Pashirova

2017-02-01

Full Text Available Over the last decade isoindigo derivatives have attracted much attention due to their high potential in pharmacy and in the chemistry of materials. In addition, isoindigo derivatives can be modified to form supramolecular structures with tunable morphologies for the use in drug delivery. Amphiphilic long-chain dialkylated isoindigos have the ability to form stable solid nanoparticles via a simple nanoprecipitation technique. Their self-assembly was investigated using tensiometry, dynamic light scattering, spectrophotometry, and fluorometry. The critical association concentrations and aggregate sizes were measured. The hydrophilic–lipophilic balance of alkylated isoindigo derivatives strongly influences aggregate morphology. In the case of short-chain dialkylated isoindigo derivatives, supramolecular polymers of 200 to 700 nm were formed. For long-chain dialkylated isoindigo derivatives, micellar aggregates of 100 to 200 nm were observed. Using micellar surfactant water-soluble forms of monosubstituted 1-hexadecylisoindigo as well as 1,1′-dimethylisoindigo were prepared for the first time. The formation of mixed micellar structures of different types in micellar anionic surfactant solutions (sodium dodecyl sulfate was determined. These findings are of practical importance and are of potential interest for the design of drug delivery systems and new nanomaterials.

Comparison of DNA Quantification Methods for Next Generation Sequencing.

Science.gov (United States)

Robin, Jérôme D; Ludlow, Andrew T; LaRanger, Ryan; Wright, Woodring E; Shay, Jerry W

2016-04-06

Next Generation Sequencing (NGS) is a powerful tool that depends on loading a precise amount of DNA onto a flowcell. NGS strategies have expanded our ability to investigate genomic phenomena by referencing mutations in cancer and diseases through large-scale genotyping, developing methods to map rare chromatin interactions (4C; 5C and Hi-C) and identifying chromatin features associated with regulatory elements (ChIP-seq, Bis-Seq, ChiA-PET). While many methods are available for DNA library quantification, there is no unambiguous gold standard. Most techniques use PCR to amplify DNA libraries to obtain sufficient quantities for optical density measurement. However, increased PCR cycles can distort the library's heterogeneity and prevent the detection of rare variants. In this analysis, we compared new digital PCR technologies (droplet digital PCR; ddPCR, ddPCR-Tail) with standard methods for the titration of NGS libraries. DdPCR-Tail is comparable to qPCR and fluorometry (QuBit) and allows sensitive quantification by analysis of barcode repartition after sequencing of multiplexed samples. This study provides a direct comparison between quantification methods throughout a complete sequencing experiment and provides the impetus to use ddPCR-based quantification for improvement of NGS quality.

Recent developments in cyanide detection: A review

Science.gov (United States)

Ma, Jian; Dasgupta, Purnendu K.

2010-01-01

The extreme toxicity of cyanide and environmental concerns from its continued industrial use continue to generate interest in facile and sensitive methods for cyanide detection. In recent years there is also additional recognition of HCN toxicity from smoke inhalation and potential use of cyanide as a weapon of terrorism. This review summarizes the literature since 2005 on cyanide measurement in different matrices ranging from drinking water and wastewater, to cigarette smoke and exhaled breath to biological fluids like blood, urine and saliva. The dramatic increase in the number of publications on cyanide measurement is indicative of the great interest in this field not only from analytical chemists, but also researchers from diverse environmental, medical, forensic and clinical arena. The recent methods cover both established and emerging analytical disciplines and include naked eye visual detection, spectrophotometry/colorimetry, capillary electrophoresis with optical absorbance detection, fluorometry, chemiluminescence, near-infrared cavity ring down spectroscopy, atomic absorption spectrometry, electrochemical methods (potentiometry/amperometry/ion chromatography-pulsed amperometry), mass spectrometry (selected ion flow tube mass spectrometry, electrospray ionization mass spectrometry, gas chromatography-mass spectrometry), gas chromatography (nitrogen phosphorus detector, electron capture detector) and quartz crystal mass monitors. PMID:20599024

Uranium hydrogeochemical and stream sediment reconnaissance data release for the Dubois NTMS Quadrangle, Idaho/Montana, including concentrations of forty-five additional elements

International Nuclear Information System (INIS)

LaDelfe, C.M.

1980-08-01

Totals of 1024 water samples and 1600 sediment samples were collected from 1669 locations in the Dubois quadrangle. Water samples were taken at streams, springs, and wells; sediment samples were collected from streams and springs. All field and analytical data are presented for waters in Appendix I-A and for sediments in I-B. All elemental analyses were performed at the LASL. Water samples were initially analyzed for uranium by fluorometry. All water samples containing more than the upper detection limit of uranium were reanalyzed by delayed neutron counting. Sediments were analyzed for uranium and thorium as well as aluminum, antimony, arsenic, barium, beryllium, bismuth, cadmium, calcium, cerium, cesium, chlorine, chromium, cobalt, copper, dysprosium, europium, gold, hafnium, iron, lanthanum, lead, lithium, lutetium, magnesium, manganese, nickel, niobium, potassium rubidium, samarium, scandium, selenium, silver, sodium, strontium, tantalum, terbium, tin, titanium, tungsten, vanadium, ytterbium, zinc and zirconium. All sediments were analyzed for uranium by delayed-neutron counting. Other elemental concentrations in sediments were determined by neutron-activation analysis for 30 elements, by x-ray fluorescence for 12 elements, and by arc-source emission spectrography for 2 elements. Analytical results for sediments are reported as parts per million

Apoptotic pathways of epothilone BMS 310705.

Science.gov (United States)

Uyar, Denise; Takigawa, Nagio; Mekhail, Tarek; Grabowski, Dale; Markman, Maurie; Lee, Francis; Canetta, Renzo; Peck, Ron; Bukowski, Ronald; Ganapathi, Ram

2003-10-01

BMS 310705 is a novel water-soluble analog of epothilone B currently in phase I clinical evaluation in the treatment of malignancies such as ovarian, renal, bladder, and lung carcinoma. Using an early passage cell culture model derived from the ascites of a patient clinically refractory to platinum/paclitaxel therapy, we evaluated the pathway of caspase-mediated apoptosis. Cells were treated for 1 h and subsequently evaluated for apoptosis, survival, and caspase activity. Apoptosis was determined by fluorescent microscopy. Caspase-3, -8, and -9 activities were determined by fluorometry using target tetrapeptide substrates. Mitochondrial release of cytochrome c was determined by immunoblot analysis. After treatment with BMS 310705, apoptosis was confirmed in >25% of cells at 24 h. Survival was significantly lower (P < 0.02) in cells treated with 0.05 micro M BMS 310705 vs paclitaxel. Analysis revealed an increase of caspase-9 and -3 activity; no caspase -8 activity was observed. Release of cytochrome c was detected at 12 h following treatment. SN-38 and topotecan failed to induce apoptosis. BMS 310705 induces significant apoptosis, decreases survival, and utilizes the mitochondrial-mediated pathway for apoptosis in this model.

Uranium hydrogeochemical and stream sediment reconnaissance data release for the Dubois NTMS Quadrangle, Idaho/Montana, including concentrations of forty-five additional elements

Energy Technology Data Exchange (ETDEWEB)

LaDelfe, C.M.

1980-08-01

Totals of 1024 water samples and 1600 sediment samples were collected from 1669 locations in the Dubois quadrangle. Water samples were taken at streams, springs, and wells; sediment samples were collected from streams and springs. All field and analytical data are presented for waters in Appendix I-A and for sediments in I-B. All elemental analyses were performed at the LASL. Water samples were initially analyzed for uranium by fluorometry. All water samples containing more than the upper detection limit of uranium were reanalyzed by delayed neutron counting. Sediments were analyzed for uranium and thorium as well as aluminum, antimony, arsenic, barium, beryllium, bismuth, cadmium, calcium, cerium, cesium, chlorine, chromium, cobalt, copper, dysprosium, europium, gold, hafnium, iron, lanthanum, lead, lithium, lutetium, magnesium, manganese, nickel, niobium, potassium rubidium, samarium, scandium, selenium, silver, sodium, strontium, tantalum, terbium, tin, titanium, tungsten, vanadium, ytterbium, zinc and zirconium. All sediments were analyzed for uranium by delayed-neutron counting. Other elemental concentrations in sediments were determined by neutron-activation analysis for 30 elements, by x-ray fluorescence for 12 elements, and by arc-source emission spectrography for 2 elements. Analytical results for sediments are reported as parts per million.

Red fluorescence increases with depth in reef fishes, supporting a visual function, not UV protection

Science.gov (United States)

Meadows, Melissa G.; Anthes, Nils; Dangelmayer, Sandra; Alwany, Magdy A.; Gerlach, Tobias; Schulte, Gregor; Sprenger, Dennis; Theobald, Jennifer; Michiels, Nico K.

2014-01-01

Why do some marine fishes exhibit striking patterns of natural red fluorescence? In this study, we contrast two non-exclusive hypotheses: (i) that UV absorption by fluorescent pigments offers significant photoprotection in shallow water, where UV irradiance is strongest; and (ii) that red fluorescence enhances visual contrast at depths below −10 m, where most light in the ‘red’ 600–700 nm range has been absorbed. Whereas the photoprotection hypothesis predicts fluorescence to be stronger near the surface and weaker in deeper water, the visual contrast hypothesis predicts the opposite. We used fluorometry to measure red fluorescence brightness in vivo in individuals belonging to eight common small reef fish species with conspicuously red fluorescent eyes. Fluorescence was significantly brighter in specimens from the −20 m sites than in those from −5 m sites in six out of eight species. No difference was found in the remaining two. Our results support the visual contrast hypothesis. We discuss the possible roles fluorescence may play in fish visual ecology and highlight the possibility that fluorescent light emission from the eyes in particular may be used to detect cryptic prey. PMID:25030989

Temperature, salinity and fluorometry profile data from CTD casts from the west Florida shelf during the ECOHAB program, 1998-1999 (NODC Accession 0000540)

Data.gov (United States)

National Oceanic and Atmospheric Administration, Department of Commerce — The USF, College of Marine Science, Ocean Circulation Group under the direction of Dr. R.H. Weisberg is maintaining an array of moored instruments on the west...

The influence of bromodeoxyuridine on the induction and repair of DNA double-strand breaks in glioblastoma cells

International Nuclear Information System (INIS)

Nusser, N.N.; Bartkowiak, D.; Roettinger, E.M.

2002-01-01

Aims: To examine the dose response of DNA damage and its modification by the radiosensitizer, 5-bromo-2'-deoxyuridine (BrdU). The sensitizing mechanism is analyzed with regard to its influence on the induction and repair of DNA double-strand breaks (DSBs). Material and Methods: Cells from three different human glioblastoma lines, A7, LH and U87MG, were X-irradiated with and without exposure to BrdU. DNA fragments were separated by field-inversion gel electrophoresis (FIGE) and quantified by fluorometry immediately and 24 h after irradiation. Results: In all cell lines, the dose response followed a linear-quadratic rather than a purely linear function. BrdU-treated cells exhibited a significantly higher amount of mobile DNA. In repair experiments with and without BrdU, the amount of mobile DNA fell close to control values within 24 h. Conclusions: The linear-quadratic model appropriately describes the X-ray induced fragmentation of DNA. BrdU sensitizing acts predominantly by increasing DNA fragility, and not by impairing damage repair. The amount of DSBs persistent after 24 h of repair is minimal, even after highly cytotoxic doses. However, it appears to depend on the extent of initial damage, causing sensitized cells to retain more DSBs than unsensitized cells. (orig.)

Rapid reactivation of cyanobacterial photosynthesis and migration upon rehydration of desiccated marine microbial mats

Directory of Open Access Journals (Sweden)

Arjun eChennu

2015-12-01

Full Text Available Desiccated cyanobacterial mats are the dominant biological feature in the Earth's arid zones. While the response of desiccated cyanobacteria to rehydration is well documented for terrestrial systems, information about the response in marine systems is lacking. We used high temporal resolution hyperspectral imaging, liquid chromatography, pulse-amplitude fluorometry, oxygen microsensors and confocal laser microscopy to study this response in a desiccated microbial mat from Exmouth Gulf, Australia. During the initial 15 minutes after rehydration chlorophyll a concentrations increased 2-5 fold and cyanobacterial photosynthesis was re-established. Although the mechanism behind this rapid increase of chlorophyll a remains unknown, we hypothesize that it involves resynthesis from a precursor stored in desiccated cyanobacteria. The subsequent phase (15 min – 48 h involved migration of the reactivated cyanobacteria towards the mat surface, which led, together with a gradual increase in chlorophyll a, to a further increase in photosynthesis. We conclude that the response involving an increase in chlorophyll a and recovery of photosynthetic activity within minutes after rehydration is common for cyanobacteria from desiccated mats of both terrestrial and aquatic origin. However the response of upward migration and its triggering factor appears to be mat-specific and likely linked to other factors.

Biochemical Modulation of Lipid Pathway in Microalgae Dunaliella sp. for Biodiesel Production

Directory of Open Access Journals (Sweden)

Ahmad Farhad Talebi

2015-01-01

Full Text Available Exploitation of renewable sources of energy such as algal biodiesel could turn energy supplies problem around. Studies on a locally isolated strain of Dunaliella sp. showed that the mean lipid content in cultures enriched by 200 mg L−1 myoinositol was raised by around 33% (1.5 times higher than the control. Similarly, higher lipid productivity values were achieved in cultures treated by 100 and 200 mg L−1 myoinositol. Fluorometry analyses (microplate fluorescence and flow cytometry revealed increased oil accumulation in the Nile red-stained algal samples. Moreover, it was predicted that biodiesel produced from myoinositol-treated cells possessed improved oxidative stability, cetane number, and cloud point values. From the genomic point of view, real-time analyses revealed that myoinositol negatively influenced transcript abundance of AccD gene (one of the key genes involved in lipid production pathway due to feedback inhibition and that its positive effect must have been exerted through other genes. The findings of the current research are not to interprete that myoinositol supplementation could answer all the challenges faced in microalgal biodiesel production but instead to show that “there is a there there” for biochemical modulation strategies, which we achieved, increased algal oil quantity and enhanced resultant biodiesel quality.

Biochemical Modulation of Lipid Pathway in Microalgae Dunaliella sp. for Biodiesel Production

Science.gov (United States)

Talebi, Ahmad Farhad; Tohidfar, Masoud; Mousavi Derazmahalleh, Seyedeh Mahsa; Sulaiman, Alawi; Baharuddin, Azhari Samsu; Tabatabaei, Meisam

2015-01-01

Exploitation of renewable sources of energy such as algal biodiesel could turn energy supplies problem around. Studies on a locally isolated strain of Dunaliella sp. showed that the mean lipid content in cultures enriched by 200 mg L−1 myoinositol was raised by around 33% (1.5 times higher than the control). Similarly, higher lipid productivity values were achieved in cultures treated by 100 and 200 mg L−1 myoinositol. Fluorometry analyses (microplate fluorescence and flow cytometry) revealed increased oil accumulation in the Nile red-stained algal samples. Moreover, it was predicted that biodiesel produced from myoinositol-treated cells possessed improved oxidative stability, cetane number, and cloud point values. From the genomic point of view, real-time analyses revealed that myoinositol negatively influenced transcript abundance of AccD gene (one of the key genes involved in lipid production pathway) due to feedback inhibition and that its positive effect must have been exerted through other genes. The findings of the current research are not to interprete that myoinositol supplementation could answer all the challenges faced in microalgal biodiesel production but instead to show that “there is a there there” for biochemical modulation strategies, which we achieved, increased algal oil quantity and enhanced resultant biodiesel quality. PMID:26146623

Regulation of Na+ channel inactivation by the DIII and DIV voltage-sensing domains.

Science.gov (United States)

Hsu, Eric J; Zhu, Wandi; Schubert, Angela R; Voelker, Taylor; Varga, Zoltan; Silva, Jonathan R

2017-03-06

Functional eukaryotic voltage-gated Na + (Na V ) channels comprise four domains (DI-DIV), each containing six membrane-spanning segments (S1-S6). Voltage sensing is accomplished by the first four membrane-spanning segments (S1-S4), which together form a voltage-sensing domain (VSD). A critical Na V channel gating process, inactivation, has previously been linked to activation of the VSDs in DIII and DIV. Here, we probe this interaction by using voltage-clamp fluorometry to observe VSD kinetics in the presence of mutations at locations that have been shown to impair Na V channel inactivation. These locations include the DIII-DIV linker, the DIII S4-S5 linker, and the DIV S4-S5 linker. Our results show that, within the 10-ms timeframe of fast inactivation, the DIV-VSD is the primary regulator of inactivation. However, after longer 100-ms pulses, the DIII-DIV linker slows DIII-VSD deactivation, and the rate of DIII deactivation correlates strongly with the rate of recovery from inactivation. Our results imply that, over the course of an action potential, DIV-VSDs regulate the onset of fast inactivation while DIII-VSDs determine its recovery. © 2017 Hsu et al.

Effects of extracellular modulation through hypoxia on the glucose metabolism of human breast cancer stem cells

Science.gov (United States)

Yustisia, I.; Jusman, S. W. A.; Wanandi, S. I.

2017-08-01

Cancer stem cells have been reported to maintain stemness under certain extracellular changes. This study aimed to analyze the effect of extracellular O2 level modulation on the glucose metabolism of human CD24-/CD44+ breast cancer stem cells (BCSCs). The primary BCSCs (CD24-/CD44+ cells) were cultured under hypoxia (1% O2) for 0.5, 4, 6, 24 and 48 hours. After each incubation period, HIF1α, GLUT1 and CA9 expressions, as well as glucose metabolism status, including glucose consumption, lactate production, O2 consumption and extracellular pH (pHe) were analyzed using qRT-PCR, colorimetry, fluorometry, and enzymatic reactions, respectively. Hypoxia caused an increase in HIF1α mRNA expressions and protein levels and shifted the metabolic states to anaerobic glycolysis, as demonstrated by increased glucose consumption and lactate production, as well as decreased O2 consumption and pHe. Furthermore, we demonstrated that GLUT1 and CA9 mRNA expressions simultaneously increased, in line with HIF1α expression. In conclusion, modulation of the extracellular environment of human BCSCs through hypoxia shifedt the metabolic state of BCSCs to anaerobic glycolysis, which might be associated with GLUT1 and CA9 expressions regulated by HIFlα transcription factor.

USE OF PULSE-AMPLITUDE-MODULATED FLUORESCENCE TO ASSESS THE PHYSIOLOGICAL STATUS OF CLADOPHORA SP. ALONG A WATER QUALITY GRADIENT(1).

Science.gov (United States)

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.

Rapid Reactivation of Cyanobacterial Photosynthesis and Migration upon Rehydration of Desiccated Marine Microbial Mats

KAUST Repository

Chennu, Arjun

2015-12-24

Desiccated cyanobacterial mats are the dominant biological feature in the Earth’s arid zones. While the response of desiccated cyanobacteria to rehydration is well-documented for terrestrial systems, information about the response in marine systems is lacking. We used high temporal resolution hyperspectral imaging, liquid chromatography, pulse-amplitude fluorometry, oxygen microsensors, and confocal laser microscopy to study this response in a desiccated microbial mat from Exmouth Gulf, Australia. During the initial 15 min after rehydration chlorophyll a concentrations increased 2–5 fold and cyanobacterial photosynthesis was re-established. Although the mechanism behind this rapid increase of chlorophyll a remains unknown, we hypothesize that it involves resynthesis from a precursor stored in desiccated cyanobacteria. The subsequent phase (15 min–48 h) involved migration of the reactivated cyanobacteria toward the mat surface, which led, together with a gradual increase in chlorophyll a, to a further increase in photosynthesis. We conclude that the response involving an increase in chlorophyll a and recovery of photosynthetic activity within minutes after rehydration is common for cyanobacteria from desiccated mats of both terrestrial and marine origin. However, the response of upward migration and its triggering factor appear to be mat-specific and likely linked to other factors.

Entrapped Sediments as a Source of Phosphorus in Epilithic Cyanobacterial Proliferations in Low Nutrient Rivers

Science.gov (United States)

Wood, Susanna A.; Depree, Craig; Brown, Logan; McAllister, Tara; Hawes, Ian

2015-01-01

Proliferations of the benthic mat-forming cyanobacteria Phormidium have been reported in rivers worldwide. Phormidium commonly produces natural toxins which pose a health risk to animal and humans. Recent field studies in New Zealand identified that sites with Phormidium proliferations consistently have low concentrations of water column dissolved reactive phosphorus (DRP). Unlike other river periphyton, Phormidium mats are thick and cohesive, with water and fine sediment trapped in a mucilaginous matrix. We hypothesized that daytime photosynthetic activity would elevate pH inside the mats, and/or night time respiration would reduce dissolved oxygen. Either condition could be sufficient to facilitate desorption of phosphates from sediment incorporated within mats, thus allowing Phormidium to utilize it for growth. Using microelectrodes, optodes and pulse amplitude modulation fluorometry we demonstrated that photosynthetic activity results in elevated pH (>9) during daytime, and that night-time respiration causes oxygen depletion (river water and this, together with elevated concentrations of elements, including iron, suggest phosphorus release from entrapped sediment. Sequential extraction of phosphorus from trapped sediment was used to investigate the role of sediment at sites on the Mangatainoka River (New Zealand) with and without Phormidium proliferations. Deposition of fine sediment (sediment can provide a source of phosphorus to support Phormidium growth and proliferation. PMID:26479491

Air exposure of coral is a significant source of dimethylsulfide (DMS) to the atmosphere.

Science.gov (United States)

Hopkins, Frances E; Bell, Thomas G; Yang, Mingxi; Suggett, David J; Steinke, Michael

2016-10-31

Corals are prolific producers of dimethylsulfoniopropionate (DMSP). High atmospheric concentrations of the DMSP breakdown product dimethylsulfide (DMS) have been linked to coral reefs during low tides. DMS is a potentially key sulfur source to the tropical atmosphere, but DMS emission from corals during tidal exposure is not well quantified. Here we show that gas phase DMS concentrations (DMS gas ) increased by an order of magnitude when three Indo-Pacific corals were exposed to air in laboratory experiments. Upon re-submersion, an additional rapid rise in DMS gas was observed, reflecting increased production by the coral and/or dissolution of DMS-rich mucus formed by the coral during air exposure. Depletion in DMS following re-submersion was likely due to biologically-driven conversion of DMS to dimethylsulfoxide (DMSO). Fast Repetition Rate fluorometry showed downregulated photosynthesis during air exposure but rapid recovery upon re-submersion, suggesting that DMS enhances coral tolerance to oxidative stress during a process that can induce photoinhibition. We estimate that DMS emission from exposed coral reefs may be comparable in magnitude to emissions from other marine DMS hotspots. Coral DMS emission likely comprises a regular and significant source of sulfur to the tropical marine atmosphere, which is currently unrecognised in global DMS emission estimates and Earth System Models.

Synergistic effects of diuron and sedimentation on photosynthesis and survival of crustose coralline algae

International Nuclear Information System (INIS)

Harrington, Lindsay; Fabricius, Katharina; Eaglesham, Geoff; Negri, Andrew

2005-01-01

Effects of short-term exposure to sedimentation and diuron, separately and in combination, on the photophysiology and survival of crustose coralline algae (CCA) were examined in controlled time-course experiments, using pulse-amplitude modulation (PAM) chlorophyll fluorometry. These experiments indicated that the effects of sediments and diuron, when applied in isolation, were often reversible, with recovery time dependant upon sediment type and diuron concentration. Exposure to fine ( m' ) of photosystem II in CCA species more than exposure to the same amount of fine ( m' ) was also observed at diuron concentrations 2.9 μg L -1 . Fine estuarine sediments in combination with 0.79 μg L -1 dissolved diuron, caused yields (ΔF/F m' ) to drop by 60% compared with controls after 24 h. The combined exposure to sediments and diuron also retarded recovery, thus ΔF/F m' values were still only 60% of the controls after 9 days recovery in clean seawater. Mortality of CCA was observed in some fragments treated with combinations of sediment and diuron. Our results suggest that sediment deposition and exposure to diuron can negatively affect the photosynthetic activity of CCA, with sedimentation stress being significantly enhanced by the presence of trace concentrations of diuron

Flow enhances photosynthesis in marine benthic autotrophs by increasing the efflux of oxygen from the organism to the water.

Science.gov (United States)

Mass, Tali; Genin, Amatzia; Shavit, Uri; Grinstein, Mor; Tchernov, Dan

2010-02-09

Worldwide, many marine coastal habitats are facing rapid deterioration due in part to human-driven changes in habitat characteristics, including changes in flow patterns, a factor known to greatly affect primary production in corals, algae, and seagrasses. The effect of flow traditionally is attributed to enhanced influx of nutrients and dissolved inorganic carbon (DIC) across the benthic boundary layer from the water to the organism however, here we report that the organism's photosynthetic response to changes in the flow is nearly instantaneous, and that neither nutrients nor DIC limits this rapid response. Using microelectrodes, dual-pulse amplitude-modulated fluorometry, particle image velocimetry, and real time mass-spectrometry with the common scleractinian coral Favia veroni, the alga Gracilaria cornea, and the seagrass Halophila stipulacea, we show that this augmented photosynthesis is due to flow-driven enhancement of oxygen efflux from the organism to the water, which increases the affinity of the RuBisCO to CO(2). No augmentation of photosynthesis was found in the absence of flow or when flow occurred, but the ambient concentration of oxygen was artificially elevated. We suggest that water motion should be considered a fundamental factor, equivalent to light and nutrients, in determining photosynthesis rates in marine benthic autotrophs.

Rapid Reactivation of Cyanobacterial Photosynthesis and Migration upon Rehydration of Desiccated Marine Microbial Mats

KAUST Repository

Chennu, Arjun; Grinham, Alistair; Polerecky, Lubos; de Beer, Dirk; Alnajjar, Mohammad Ahmad

2015-01-01

Desiccated cyanobacterial mats are the dominant biological feature in the Earth’s arid zones. While the response of desiccated cyanobacteria to rehydration is well-documented for terrestrial systems, information about the response in marine systems is lacking. We used high temporal resolution hyperspectral imaging, liquid chromatography, pulse-amplitude fluorometry, oxygen microsensors, and confocal laser microscopy to study this response in a desiccated microbial mat from Exmouth Gulf, Australia. During the initial 15 min after rehydration chlorophyll a concentrations increased 2–5 fold and cyanobacterial photosynthesis was re-established. Although the mechanism behind this rapid increase of chlorophyll a remains unknown, we hypothesize that it involves resynthesis from a precursor stored in desiccated cyanobacteria. The subsequent phase (15 min–48 h) involved migration of the reactivated cyanobacteria toward the mat surface, which led, together with a gradual increase in chlorophyll a, to a further increase in photosynthesis. We conclude that the response involving an increase in chlorophyll a and recovery of photosynthetic activity within minutes after rehydration is common for cyanobacteria from desiccated mats of both terrestrial and marine origin. However, the response of upward migration and its triggering factor appear to be mat-specific and likely linked to other factors.

Changes in intestinal intraepithelial lymphocytes due to local irradiation of a portion of the maxilla in mice

Energy Technology Data Exchange (ETDEWEB)

Satoh, Daigo; Tamazawa, Ken; Yosue, Takashi; Sakai, Yasuo; Suzuki, Masayuki [Nihon Univ., Tokyo (Japan). School of Dentistry; Arai, Chiaki; Inagaki, Hiroyuki

2000-05-01

We have been investigating the changes of lymphocyte subsets in the immune organs after head and neck irradiation. The present study investigated the influence due to local irradiation (10 Gy) of a portion of the maxilla on intestinal intraepithelial lymphocytes (i-IEL) in mice. We analyzed the percentage of subpopulations in i-IEL following irradiation using two-color fluorometry (anti-TCR{alpha}{beta}, TCR{gamma}{delta}, CD4 and CD8 monoclonal antibodies), and the outcome was compared with that obtained from non-irradiation groups. The result was that the percentage of {alpha}{beta}T cells, {gamma}{delta}T cells, CD4{sup +}SPT cells and CD8{sup +}SPT cells did not show any significant change and the fact differed from that in spleen and thymus where the percentage of these subpopulations was significantly changed. Although spleen and thymus reflect damaged lymphocytes that had circulated through the irradiation field, the result indicates that the balance among i-IEL subsets does not change after irradiation. The reason that equilibrium of i-IEL subsets remains stable seems related to localization of the small intestine and radioresistance. It was also indicated that the effect on the oral epithelium in the irradiation field reflects the percentages of i-IEL subsets. (author)

Hazard and risk of herbicides for marine microalgae

International Nuclear Information System (INIS)

Sjollema, Sascha B.; MartínezGarcía, Gema; Geest, Harm G. van der; Kraak, Michiel H.S.; Booij, Petra; Vethaak, A. Dick; Admiraal, Wim

2014-01-01

Due to their specific effect on photosynthesis, herbicides pose a potential threat to coastal and estuarine microalgae. However, comprehensive understanding of the hazard and risk of these contaminants is currently lacking. Therefore the aim of the present study was to investigate the toxic effects of four ubiquitous herbicides (atrazine, diuron, Irgarol ® 1051 and isoproturon) and herbicide mixtures on marine microalgae. Using a Pulse Amplitude Modulation (PAM) fluorometry based bioassay we demonstrated a clear species and herbicide specific toxicity and showed that the current environmental legislation does not protect algae sufficiently against diuron and isoproturon. Although a low actual risk of herbicides in the field was demonstrated, monitoring data revealed that concentrations occasionally reach potential effect levels. Hence it cannot be excluded that herbicides contribute to observed changes in phytoplankton species composition in coastal waters, but this is likely to occur only occasionally. - Highlights: • The hazard of herbicides for microalgae is compound and species specific. • In general a low risk although occasional potential effect levels are reached. • Current legislation does not protect marine microalgae sufficiently. - The hazard of herbicides in the coastal waters is compound and species specific and although the general risk in the field is low, occasionally potential effect levels are reached

Do plastic particles affect microalgal photosynthesis and growth?

Science.gov (United States)

Sjollema, Sascha B; Redondo-Hasselerharm, Paula; Leslie, Heather A; Kraak, Michiel H S; Vethaak, A Dick

2016-01-01

The unbridled increase in plastic pollution of the world's oceans raises concerns about potential effects these materials may have on microalgae, which are primary producers at the basis of the food chain and a major global source of oxygen. Our current understanding about the potential modes and mechanisms of toxic action that plastic particles exert on microalgae is extremely limited. How effects might vary with particle size and the physico-chemical properties of the specific plastic material in question are equally unelucidated, but may hold clues to how toxicity, if observed, is exerted. In this study we selected polystyrene particles, both negatively charged and uncharged, and three different sizes (0.05, 0.5 and 6μm) for testing the effects of size and material properties. Microalgae were exposed to different polystyrene particle sizes and surface charges for 72h. Effects on microalgal photosynthesis and growth were determined by pulse amplitude modulation fluorometry and flow cytometry, respectively. None of the treatments tested in these experiments had an effect on microalgal photosynthesis. Microalgal growth was negatively affected (up to 45%) by uncharged polystyrene particles, but only at high concentrations (250mg/L). Additionally, these adverse effects were demonstrated to increase with decreasing particle size. Copyright © 2015 Elsevier B.V. All rights reserved.

Laser spectroscopy on organic molecules.

Science.gov (United States)

Imasaka, T

1996-06-01

Various laser spectrometric methods have been developed until now. Especially, laser fluorometry is most sensitive and is frequently combined with a separation technique such as capillary electrophoresis. For non-fluorescent compounds, photothermal spectrometry may be used instead. A diode laser is potentially useful for practical trace analysis, because of its low cost and long-term trouble-free operation. On the other hand, monochromaticity of the laser is essential in high-resolution spectrometry, e.g. in low temperature spectrometry providing a very sharp spectral feature. Closely-related compounds such as isomers can easily be differentiated, and information for assignment is obtained from the spectrum. Multiphoton ionization mass spectrometry is useful for soft ionization, providing additional information concerned with molecular weight and chemical structure. A short laser pulse with a sufficient energy is suitable for rapid heating of the solid surface. A matrix-assisted laser desorption/ion-ization technique is recently employed for introduction of a large biological molecule into a vacuum for mass analysis. In the future, laser spectrometry will be developed by a combination with state-of-the-art laser technology. In the 21st century, new laser spectrometry will be developed, which may be based on revolutionary ideas or unexpected discoveries. Such studies will open new frontiers in analytical laser spectroscopy.

Identification and Characterization of Amlexanox as a G Protein-Coupled Receptor Kinase 5 Inhibitor

Directory of Open Access Journals (Sweden)

Kristoff T. Homan

2014-10-01

Full Text Available G protein-coupled receptor kinases (GRKs have been implicated in human diseases ranging from heart failure to diabetes. Previous studies have identified several compounds that selectively inhibit GRK2, such as paroxetine and balanol. Far fewer selective inhibitors have been reported for GRK5, a target for the treatment of cardiac hypertrophy, and the mechanism of action of reported compounds is unknown. To identify novel scaffolds that selectively inhibit GRK5, a differential scanning fluorometry screen was used to probe a library of 4480 compounds. The best hit was amlexanox, an FDA-approved anti-inflammatory, anti-allergic immunomodulator. The crystal structure of amlexanox in complex with GRK1 demonstrates that its tricyclic aromatic ring system forms ATP-like interactions with the hinge of the kinase domain, which is likely similar to how this drug binds to IκB kinase ε (IKKε, another kinase known to be inhibited by this compound. Amlexanox was also able to inhibit myocyte enhancer factor 2 transcriptional activity in neonatal rat ventricular myocytes in a manner consistent with GRK5 inhibition. The GRK1 amlexanox structure thus serves as a springboard for the rational design of inhibitors with improved potency and selectivity for GRK5 and IKKε.

Mitochondrial function and tissue vitality: bench-to-bedside real-time optical monitoring system

Science.gov (United States)

Mayevsky, Avraham; Walden, Raphael; Pewzner, Eliyahu; Deutsch, Assaf; Heldenberg, Eitan; Lavee, Jacob; Tager, Salis; Kachel, Erez; Raanani, Ehud; Preisman, Sergey; Glauber, Violete; Segal, Eran

2011-06-01

Background: The involvement of mitochondria in pathological states, such as neurodegenerative diseases, sepsis, stroke, and cancer, are well documented. Monitoring of nicotinamide adenine dinucleotide (NADH) fluorescence in vivo as an intracellular oxygen indicator was established in 1950 to 1970 by Britton Chance and collaborators. We use a multiparametric monitoring system enabling assessment of tissue vitality. In order to use this technology in clinical practice, the commercial developed device, the CritiView (CRV), is tested in animal models as well as in patients. Methods and Results: The new CRV enables the optical monitoring of four different parameters, representing the energy balance of various tissues in vivo. Mitochondrial NADH is measured by surface fluorometry/reflectometry. In addition, tissue microcirculatory blood flow, tissue reflectance and oxygenation are measured as well. The device is tested both in vitro and in vivo in a small animal model and in preliminary clinical trials in patients undergoing vascular or open heart surgery. In patients, the monitoring is started immediately after the insertion of a three-way Foley catheter (urine collection) to the patient and is stopped when the patient is discharged from the operating room. The results show that monitoring the urethral wall vitality provides information in correlation to the surgical procedure performed.

Canopy uptake of atmospheric N deposition at a conifer forest: part I -canopy N budget, photosynthetic efficiency and net ecosystem exchange

International Nuclear Information System (INIS)

Sievering, H.; Tomaszewski, T.; Torizzo, J.

2007-01-01

Global carbon cycle assessments of anthropogenic nitrogen (N) deposition influences on carbon sequestration often assume enhanced sequestration results. This assumption was evaluated at a Rocky Mountains spruce-fir forest. Forest canopy N uptake (CNU) of atmospheric N deposition was estimated by combining event wet and throughfall N fluxes with gradient measured HNO 3 and NH 3 as well as inferred (NO x and particulate N) dry fluxes. Approximately 80% of the growing-season 3 kg N/ha total deposition is retained in canopy foliage and branches. This CNU constitutes ∼1/3 of canopy growing season new N supply at this conifer forest site. Daytime net ecosystem exchange (NEE) significantly (P = 0.006) and negatively (CO 2 uptake) correlated with CNU. Multiple regression indicates ∼20% of daytime NEE may be attributed to CNU (P < 0.02); more than soil water content. A wet deposition N-amendment study (Tomaszewski and Sievering), at canopy spruce branches, increased their growing-season CNU by 40-50% above ambient. Fluorometry and gas exchange results show N-amended spruce branches had greater photosynthetic efficiency and higher carboxylation rates than control and untreated branches. N-amended branches had 25% less photoinhibition, with a 5-9% greater proportion of foliar-N-in-Rubisco. The combined results provide, partly, a mechanistic explanation for the NEE dependence on CNU

Blood perfusion and pH monitoring in organs by laser-induced fluorescence spectroscopy

Science.gov (United States)

Vari, Sandor G.; Papazoglou, Theodore G.; Pergadia, Vani R.; Stavridi, Marigo; Snyder, Wendy J.; Papaioannou, Thanassis; Duffy, J. T.; Weiss, Andrew B.; Thomas, Reem; Grundfest, Warren S.

1994-01-01

Sensitivity of laser-induced fluorescence spectroscopy (LIFS) in detecting a change in tissue pH, and blood perfusion was determined. Rabbits were anesthetized, paralyzed, and mechanically ventilated. The arterial and venous blood supplies of the kidney were isolated and ligated to alter the perfusion. The femoral artery was cannulated to extract samples for blood gas analysis. A 308-nm XeCl was used as an excitation source. A 600 micrometers core diameter fiber was used for fluorescence acquisition, and the spectra analyzed by an optical multichannel analyzer (EG & G, OMA III). the corresponding intensity ratio R equals INADH / ICOLL was used as an index for respiratory acidosis. Blood perfusion was assessed using the following algorithm: (IELAS minus ICOLL) divided by (INADH minus ICOLL). The intensity ratio linearly decreased with the reduction of blood perfusion. When we totally occluded the artery the ratio decreased tenfold when compared to the ratio of a fully perfused kidney. Results of monitoring blood acidosis by laser-induced fluorescence spectroscopy shows a significant trend between pH and intensity ratio. Since all the slopes were negative, there is an obvious significant correlation between the pH and NADH.COLLAGEN RATIO. Blue-light-induced fluorescence measurements and ratio fluorometry is a sensitive method for monitoring blood perfusion and acidity or alkalinity of an organ.

Uranium hydrogeochemical and stream sediment reconnaissance data release for the Elk City NTMS Quadrangle, Idaho/Montana, including concentrations of forty-five additional elements

Energy Technology Data Exchange (ETDEWEB)

Broxton, D.E.; Beyth, M.

1980-07-01

Totals of 1580 water and 1720 sediment samples were collected from 1754 locations in the quadrangle. Elemental concentration, field measurement, weather, geologic, and geographic data for each sample location are listed for waters in Appendix I-A and for sediments in Appendix I-B. Uranium/thorium ratios for sediment samples are also included in Appendix I-B. All elemental analyses were performed at the LASL. Water samples were initially analyzed for uranium by fluorometry. All water samples containing more than 40 parts per billion (ppB) uranium were reanalyzed by delayed-neutron counting (DNC). A supplemental report containing the multielement analyses of water samples will be open filed in the near future. Sediments were analyzed for uranium and thorium as well as aluminum, antimony, arsenic, barium, beryllium, bismuth, cadmium, calcium, cerium, cesium, chlorine, chromium, cobalt, copper, dysprosium, europium, gold, hafnium, iron, lanthanum, lead, lithium, lutetium, magnesium, manganese, nickel, niobium, potassium, rubidium, samarium, selenium, scandium, silver, sodium, strontium, tantalum, terbium, tin, titanium, tungsten, vanadium, ytterbium, zinc, and zirconium. Basic statistics for 40 of these elements are presented. All sediments were analyzed for uranium by delayed-neutron counting. Other elemental concentrations in sediments were determined by neutron-activation analysis for 30 elements, by x-ray fluorescence for 12 elements, and by arc-source emission spectrography for 2 elements. Analytical results for sediments are reported as parts per million.

Photosynthetic and cellular toxicity of cadmium in Chlorella vulgaris.

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Ou-Yang, Hui-Ling; Kong, Xiang-Zhen; Lavoie, Michel; He, Wei; Qin, Ning; He, Qi-Shuang; Yang, Bin; Wang, Rong; Xu, Fu-Liu

2013-12-01

The toxic effects of cadmium (Cd) on the green alga Chlorella vulgaris were investigated by following the response to Cd of various toxicity endpoints (cell growth, cell size, photochemical efficiency of PSII in the light or Φ(PSII), maximal photochemical efficiency or Fv/Fm, chlorophyll a fluorescence, esterase activity, and cell viability). These toxicity endpoints were studied in laboratory batch cultures of C. vulgaris over a long-term 96-h exposure to different Cd concentrations using flow cytometry and pulse amplitude modulated fluorometry. The sequence of sensitivity of these toxicity endpoints was: cell yield > Φ(PSII) ≈ esterase activity > Fv/Fm > chlorophyll a fluorescence ≈ cell viability. It is shown that cell apoptosis or cell death only accounted for a minor part of the reduction in cell yield even at very high algistatic free Cd²⁺ concentrations, and other mechanisms such as blocked cell divisions are major contributors to cell yield inhibition. Furthermore, cadmium may affect both the electron donors and acceptors of the electron transport chain at high free Cd²⁺ concentration. Finally, the resistance of cells to cell death was size-dependent; medium-sized cells had the highest toxicity threshold. The present study brings new insights into the toxicity mechanisms of Cd in C. vulgaris and provides a detailed comparison of the sensitivity of various Cd toxicity endpoints. © 2013 SETAC.

Cytomegalovirus and Toxoplasma gondii seroprevalence in a Brazilian liver transplant waiting

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R. A. M. B. Almeida

2007-01-01

Full Text Available Cytomegalovirus (CMV disease is a major cause of morbidity and mortality in solid organ transplantation. Disseminated toxoplasmosis after liver transplantation is a rare but fatal event. Serologic screening of the donor and the recipient is essential to prophylactic management, early diagnosis and therapeutic strategies to minimize the consequences of these infections. The aim of the present study was to determine the seroprevalence of CMV and Toxoplasma gondii (TG in a Brazilian liver transplant waiting list (LTWL. Serological data were collected from 44 candidates on the LTWL between May 2003 and November 2004. Serological investigation of antibodies IgM and IgG against CMV (anti-CMV and TG (anti-T. gondii was performed using fluorometry commercial kits. IgG anti-CMV was positive in 37 patients (94.9% out of 39 available results. There were not IgM anti-CMV positive results. Out of 36 analyzed patients, 22 (61.1% presented positive IgG anti-T. gondii and none had positive IgM anti-T. gondii. The high CMV seroprevalence among our LTWL reinforces the need for appropriate protocols to avoid related complications, like reactivation and superinfection by CMV. Environmental and drug prophylactic strategies against primary infection and reactivation, as well as early diagnosis and treatment of toxoplasmosis complications, are essential for the good outcome of transplant patients.

Ecotoxicology of bromoacetic acid on estuarine phytoplankton

International Nuclear Information System (INIS)

Gordon, Ana R.; Richardson, Tammi L.; Pinckney, James L.

2015-01-01

Bromoacetic acid is formed when effluent containing chlorine residuals react with humics in natural waters containing bromide. The objective of this research was to quantify the effects of bromoacetic acid on estuarine phytoplankton as a proxy for ecosystem productivity. Bioassays were used to measure the EC 50 for growth in cultured species and natural marine communities. Growth inhibition was estimated by changes in chlorophyll a concentrations measured by fluorometry and HPLC. The EC 50 s for cultured Thalassiosira pseudonana were 194 mg L −1 , 240 mg L −1 for Dunaliella tertiolecta and 209 mg L −1 for Rhodomonas salina. Natural phytoplankton communities were more sensitive to contamination with an EC 50 of 80 mg L −1 . Discriminant analysis suggested that bromoacetic acid additions cause an alteration of phytoplankton community structure with implications for higher trophic levels. A two-fold EC 50 decrease in mixed natural phytoplankton populations affirms the importance of field confirmation for establishing water quality criteria. - Highlights: • Bromoacetic acid exposure resulted in lethal impacts to estuarine phytoplankton. • Cultured phytoplankton were less sensitive to bromoacetic acid than natural communities. • Lab results should be confirmed with field experiments whenever possible. - The toxicology of haloacetic acids has been studied in freshwater ecosystems, and urbanization of the coastal zone is making effects in marine ecosystems equally relevant.

The review of identification and assay methods of β-blockers

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Ольга Олександрівна Віслоус

2015-10-01

Full Text Available Every year the number of β-blockers on the pharmaceutical market is increasing, requiring systematization of their standardization methods.Aim of research. The aim of our research is to study literature data about identification and assay methods of β-blockers with different direction of action – selective (praktolol, metoprolol, atenolol, acebutolol, betaxolol, bevantolol, bisoprolol, celiprolol, esmolol, epanolol, esatenolol, nebivolol, Talinolol, non-selective (alprenolol, Oxprenololum, pindolol, propranolol, timolol, sotalol, nadolol, mepindolol, karteol, tertatolol, bopindolol, bupranolol, penbutolol, kloranolol and combined (labetalol, carvedilol.Methods. The analytical review of literature sources about β-blockers analysis by physical, chemical, and physicochemical methods.Results. After literature sources’ analyzing it was found that physical and physicochemical constants are basically used for β-blockers pharmacopoeial analysis; both physicochemical values and chemical reactions are used in forensic analysis, resulting in the article.It was founded that titration methods, mostly acid-base titration method, are used for β-blockers assay in the analysis of substances. For β-blockers detection in biological fluids and dosage forms, active pharmaceutical ingredients and metabolites mixture separation one should prefer physicochemical methods, such as gas chromatography and liquid chromatography, absorption UV-Visible spectroscopy, fluorometry, etc.Conclusion. The results have shown can be used for the further search of the identification and assay optimal methods of β-blockers both pure and mixed with other active substances and excipients

Dynamic fluorescence studies of beta-glycosidase mutants from Sulfolobus solfataricus: effects of single mutations on protein thermostability.

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Bismuto, Ettore; Febbraio, Ferdinando; Limongelli, Simona; Briante, Raffaella; Nucci, Roberto

2003-04-01

Multiple sequence alignment on 73 proteins belonging to glycosyl hydrolase family 1 reveals the occurrence of a segment (83-124) in the enzyme sequences from hyperthermophilic archaea bacteria, which is absent in all the mesophilic members of the family. The alignment of the known three-dimensional structures of hyperthermophilic glycosidases with the known ones from mesophilic organisms shows a similar spatial organizations of beta-glycosidases except for this sequence segment whose structure is located on the external surface of each of four identical subunits, where it overlaps two alpha-helices. Site-directed mutagenesis substituting N97 or S101 with a cysteine residue in the sequence of beta-glycosidase from hyperthermophilic archaeon Sulfolobus solfataricus caused some changes in the structural and dynamic properties as observed by circular dichroism in far- and near-UV light, as well as by frequency domain fluorometry, with a simultaneous loss of thermostability. The results led us to hypothesize an important role of the sequence segment present only in hyperthermophilic beta-glycosidases, in the thermal adaptation of archaea beta-glycosidases. The thermostabilization mechanism could occur as a consequence of numerous favorable ionic interactions of the 83-124 sequence with the other part of protein matrix that becomes more rigid and less accessible to the insult of thermal-activated solvent molecules. Copyright 2003 Wiley-Liss, Inc.

Cryobiology of coral fragments.

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Hagedorn, Mary; Farrell, Ann; Carter, Virginia L

2013-02-01

Around the world, coral reefs are dying due to human influences, and saving habitat alone may not stop this destruction. This investigation focused on the biological processes that will provide the first steps in understanding the cryobiology of whole coral fragments. Coral fragments are a partnership of coral tissue and endosymbiotic algae, Symbiodinium sp., commonly called zooxanthellae. These data reflected their separate sensitivities to chilling and a cryoprotectant (dimethyl sulfoxide) for the coral Pocillopora damicornis, as measured by tissue loss and Pulse Amplitude Modulated fluorometry 3weeks post-treatment. Five cryoprotectant treatments maintained the viability of the coral tissue and zooxanthellae at control values (1M dimethyl sulfoxide at 1.0, 1.5 and 2.0h exposures, and 1.5M dimethyl sulfoxide at 1.0 and 1.5h exposures, P>0.05, ANOVA), whereas 2M concentrations did not (Pzooxanthellae. During the winter when the fragments were chilled, the coral tissue remained relatively intact (∼25% loss) post-treatment, but the zooxanthellae numbers in the tissue declined after 5min of chilling (Pzooxanthellae numbers declined in response to chilling alone (P0.05, ANOVA), but it did not protect against the loss of zooxanthellae (Pzooxanthellae are the most sensitive element in the coral fragment complex and future cryopreservation protocols must be guided by their greater sensitivity. Copyright © 2012 Elsevier Inc. All rights reserved.

Thermometry in dielectrophoresis chips for contact-free cell handling

International Nuclear Information System (INIS)

Jaeger, M S; Mueller, T; Schnelle, T

2007-01-01

Cell biology applications, protocols in immunology and stem cell research, require that individual cells are handled under strict control of their contacts to other cells or synthetic surfaces. Dielectrophoresis (DEP) in microfluidic chips is an established technique to investigate, group, wash, cultivate and sort cells contact-free under physiological conditions: microelectrode octode cages, versatile dielectrophoretic elements energized with radio frequency electric fields, stably trap single cells or cellular aggregates. For medical applications and cell cultivation, possible side effects of the dielectrophoretic manipulation, such as membrane polarization and Joule heating, have to be quantified. Therefore, we characterized the electric field-induced warming in dielectrophoretic cages using ohmic resistance measurements, fluorometry, liquid crystal beads, infra-red thermography and bubble size thermometry. We compare the results of these techniques with respect to the influences of voltage, electric conductivity of buffer, frequency, cage size and electrode surface. We conclude that in the culture medium thermal effects may be neglected if low voltages and an electric field-reducing phase pattern are used. Our experimental results provide explicit values for estimating the thermal effect on dielectrophoretically caged cells and show that Joule heating is best minimized by optimizing the cage geometry and reducing the buffer conductivity. The results may additionally serve to evaluate and improve theoretical predictions on field-induced effects. Based on present-day chip processing possibilities, DEP is well suited for the manipulation of cells

Prolyl oligopeptidase and dipeptidyl peptidase II/dipeptidyl peptidase IV ratio in the cerebrospinal fluid in Parkinson's disease: historical overview and future prospects.

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Nagatsu, Toshiharu

2017-06-01

Prolyl oligopeptidase (also named prolyl endopeptidase; PREP) hydrolyzes the Pro-Xaa bonds of biologically active oligopeptides on their carboxyl side. In 1987, we detected PREP activity in human cerebrospinal fluid (CSF) using highly sensitive liquid chromatography-fluorometry with succinyl-Gly-Pro-4-methyl-coumarin amide as a new synthetic substrate, and found a marked decrease in its activity in the cerebrospinal fluid (CSF) from patients with Parkinson's disease (PD) as compared with its level in control patients without neurological diseases. In 2013, Hannula et al. found co-localization of PREP with α-synuclein in the postmortem PD brain. Several recent studies also suggest that the level of PREP in the brain of PD patients may be related to dopamine (DA) cell death via promotion of α-synuclein oligomerization and that inhibitors of PREP may play a neuroprotective role in PD. Although the relationship between another family of prolyl oligopeptidase enzymes, dipeptidyl peptidase II (DPP II) and dipeptidyl peptidase IV (DPP IV), and α-synuclein in the PD brain is not yet clear, we found that the DPP II activity/DPP IV activity ratio in the CSF was significantly increased in PD patients. This review discusses the possibility of PREP as well as the DPP II/DPP IV ratio in the CSF as potential biomarkers of PD.

Magnetic nanoparticles grafted with β-cyclodextrin for solid-phase extraction of 5-hydroxy-3-indole acetic acid

International Nuclear Information System (INIS)

Ahmed, Gaber Hashem Gaber; Laíño, Rosana Badía; Calzón, Josefa Angela García; García, Marta Elena Díaz

2014-01-01

We describe the synthesis of β-cyclodextrin modified magnetic nanoparticles (CD-mNPs) as a material for solid-phase extraction of the cancer biomarker 5-hydroxy-indole-3-acetic acid (5-HIAA) from urine. The CD-mNPs were characterized by TEM, FTIR, and XRD, and the kinetics and adsorption isotherms were studied. The strong interaction between the CD-mNPs and 5-HIAA is the main driving force for recognition and extraction, while the magnetic core of the NPs allows their separation from the sample matrix. Recovery of 5-HIAA from the adsorbent using an adequate solvent regenerated the adsorbent for further use. 5-HIAA was then quantified by fluorometry of its complex with β-CD. The method works in the 1 × 10 −7 to 1 × 10 −5 mol L −1 (R 2 0.9982–0.9996) concentration range, and the limits of detection (3σ) and quantification (10 σ) of the method are 1.2 × 10 −8 mol L −1 and 4.01 × 10 −8 mol L −1 5-HIAA, respectively. The recovery of 5-HIAA from urine samples spiked with 5-HIAA in three concentrations (1.4 × 10 −6 , 4.50 × 10 −6 and 1.0 × 10 −5 mol L −1 ) are within 63 ± 3 %. (author)

Uranium concentrations in stream waters and sediments from selected sites in the eastern Seward Peninsula, Koyukuk, and Charley River areas, and across South-Central Alaska

International Nuclear Information System (INIS)

Sharp, R.R. Jr.; Hill, D.E.

1978-04-01

During the summer of 1975, a 6-week reconnaissance was conducted in widespread areas of Alaska as part of the National Uranium Resource Evaluation (NURE) program; Water, stream sediment, and bedrock samples were taken from the eastern Seward Peninsula, from north of Koyukuk River, from the Charley River area, and from across south central Alaska. This report contains the LASL uranium determinations resulting from fluorometric analysis of the water samples and delayed-neutron counting of the stream sediment samples. Results of total uranium for 611 water and 641 sediment samples, from 691 stream locations, are presented. Overlays showing the numbered sample locations and graphically portraying the concentrations of uranium in water and stream sediment samples, at 1:250,000 scale for use with existing National Topographic Map Series (NTMS) sheets and published geologic maps, are provided as plates. The main purposes of this work are to make the uranium data available to the public in the standard computer format used in the NURE Hydrogeochemical and Stream Sediment Reconnaissance (i.e., with a DOE sample number giving the latitude and longitude of each sample location) and to provide uranium concentration overlays at the standard scale of 1:250,000 adopted by the DOE for the NURE program. It also allows a plausible explanation of differences between the uranium values for sediment as determined by acid dissolution/extraction/fluorometry and by delayed-neutron counting that were noted in the earlier report

Codon optimisation to improve expression of a Mycobacterium avium ssp. paratuberculosis-specific membrane-associated antigen by Lactobacillus salivarius.

Science.gov (United States)

Johnston, Christopher; Douarre, Pierre E; Soulimane, Tewfik; Pletzer, Daniel; Weingart, Helge; MacSharry, John; Coffey, Aidan; Sleator, Roy D; O'Mahony, Jim

2013-06-01

Subunit and DNA-based vaccines against Mycobacterium avium ssp. paratuberculosis (MAP) attempt to overcome inherent issues associated with whole-cell formulations. However, these vaccines can be hampered by poor expression of recombinant antigens from a number of disparate hosts. The high G+C content of MAP invariably leads to a codon bias throughout gene expression. To investigate if the codon bias affects recombinant MAP antigen expression, the open reading frame of a MAP-specific antigen MptD (MAP3733c) was codon optimised for expression against a Lactobacillus salivarius host. Of the total 209 codons which constitute MAP3733c, 172 were modified resulting in a reduced G+C content from 61% for the native gene to 32.7% for the modified form. Both genes were placed under the transcriptional control of the PnisA promoter; allowing controlled heterologous expression in L. salivarius. Expression was monitored using fluorescence microscopy and microplate fluorometry via GFP tags translationally fused to the C-termini of the two MptD genes. A > 37-fold increase in expression was observed for the codon-optimised MAP3733synth variant over the native gene. Due to the low cost and improved expression achieved, codon optimisation significantly improves the potential of L. salivarius as an oral vaccine stratagem against Johne's disease. © 2013 Federation of European Microbiological Societies. Published by John Wiley & Sons Ltd. All rights reserved.

Mechanism of the negative force-frequency relationship in physiologically intact rat ventricular myocardium. Studies by intracellular Ca2+ monitor with iodo-1 and by 31P-nuclear magnetic resonance spectroscopy

International Nuclear Information System (INIS)

Morii, Isao; Kihara, Yasuki; Sasayama, Shigetake; Konishi, Takashi; Inubushi, Toshiro.

1996-01-01

We studied the subcellular mechanisms of the negative force-frequency relationship in rat myocardium by measuring intracellular Ca 2+ transients by indo-1 fluorometry and intracellular pH (pH i ) and phosphate compounds with 31 P-nuclear magnetic resonance (NMR). The data were compared with those from guinea pig hearts, which show a positive force-frequency relationship. By increasing the pacing rate from 3 Hz to 5 Hz, the peak positive first derivative of left ventricular pressure (LVdP/dt) in rat heart decreased by 10±1% (n=6). In contrast to this negative inotropic response, simultaneously measured peak Ca 2+ transients increased by 6±1%. Guinea pig heart (n=6) showed an increase in peak positive LVdP/dt (33±1%) which was associated with an increase in peak Ca 2+ transients (8±1%). Under equivalent experimental conditions in an NMR spectrometer, this increase in the pacing rate did not affect intracellular levels of phosphate compounds in either rat (n=6) or guinea pig heart (n=6). In contrast, pH i showed a decrease of 0.031±0.006 pH units in rat heart, while no changes were observed in guinea pig heart. These results suggest that in physiological rat myocardium, pH i is susceptible to changes in the stimulus frequency and may affect the Ca 2+ -responsiveness of contractile proteins, which results in the negative force-frequency relationship. (author)

Voltage-sensing domain mode shift is coupled to the activation gate by the N-terminal tail of hERG channels.

Science.gov (United States)

Tan, Peter S; Perry, Matthew D; Ng, Chai Ann; Vandenberg, Jamie I; Hill, Adam P

2012-09-01

Human ether-a-go-go-related gene (hERG) potassium channels exhibit unique gating kinetics characterized by unusually slow activation and deactivation. The N terminus of the channel, which contains an amphipathic helix and an unstructured tail, has been shown to be involved in regulation of this slow deactivation. However, the mechanism of how this occurs and the connection between voltage-sensing domain (VSD) return and closing of the gate are unclear. To examine this relationship, we have used voltage-clamp fluorometry to simultaneously measure VSD motion and gate closure in N-terminally truncated constructs. We report that mode shifting of the hERG VSD results in a corresponding shift in the voltage-dependent equilibrium of channel closing and that at negative potentials, coupling of the mode-shifted VSD to the gate defines the rate of channel closure. Deletion of the first 25 aa from the N terminus of hERG does not alter mode shifting of the VSD but uncouples the shift from closure of the cytoplasmic gate. Based on these observations, we propose the N-terminal tail as an adaptor that couples voltage sensor return to gate closure to define slow deactivation gating in hERG channels. Furthermore, because the mode shift occurs on a time scale relevant to the cardiac action potential, we suggest a physiological role for this phenomenon in maximizing current flow through hERG channels during repolarization.

KCNQ1 channel modulation by KCNE proteins via the voltage-sensing domain.

Science.gov (United States)

Nakajo, Koichi; Kubo, Yoshihiro

2015-06-15

The gating of the KCNQ1 potassium channel is drastically regulated by auxiliary subunit KCNE proteins. KCNE1, for example, slows the activation kinetics of KCNQ1 by two orders of magnitude. Like other voltage-gated ion channels, the opening of KCNQ1 is regulated by the voltage-sensing domain (VSD; S1-S4 segments). Although it has been known that KCNE proteins interact with KCNQ1 via the pore domain, some recent reports suggest that the VSD movement may be altered by KCNE. The altered VSD movement of KCNQ1 by KCNE proteins has been examined by site-directed mutagenesis, the scanning cysteine accessibility method (SCAM), voltage clamp fluorometry (VCF) and gating charge measurements. These accumulated data support the idea that KCNE proteins interact with the VSDs of KCNQ1 and modulate the gating of the KCNQ1 channel. In this review, we will summarize recent findings and current views of the KCNQ1 modulation by KCNE via the VSD. In this context, we discuss our recent findings that KCNE1 may alter physical interactions between the S4 segment (VSD) and the S5 segment (pore domain) of KCNQ1. Based on these findings from ourselves and others, we propose a hypothetical mechanism for how KCNE1 binding alters the VSD movement and the gating of the channel. © 2015 The Authors. The Journal of Physiology © 2015 The Physiological Society.

Highly selective apo-arginase based method for sensitive enzymatic assay of manganese (II) and cobalt (II) ions

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Stasyuk, Nataliya; Gayda, Galina; Zakalskiy, Andriy; Zakalska, Oksana; Errachid, Abdelhamid; Gonchar, Mykhailo

2018-03-01

A novel enzymatic method of manganese (II) and cobalt (II) ions assay, based on using apo-enzyme of Mn2 +-dependent recombinant arginase I (arginase) and 2,3-butanedione monoxime (DMO) as a chemical reagent is proposed. The principle of the method is the evaluation of the activity of L-arginine-hydrolyzing of arginase holoenzyme after the specific binding of Mn2 + or Co2 + with apo-arginase. Urea, which is the product of enzymatic hydrolysis of L-arginine (Arg), reacts with DMO and the resulted compound is detected by both fluorometry and visual spectrophotometry. Thus, the content of metal ions in the tested samples can be determined by measuring the level of urea generated after enzymatic hydrolysis of Arg by reconstructed arginase holoenzyme in the presence of tested metal ions. The linearity range of the fluorometric apo-arginase-DMO method in the case of Mn2 + assay is from 4 pM to 1.10 nM with a limit of detection of 1 pM Mn2 +, whereas the linearity range of the present method in the case of Co2 + assay is from 8 pM to 45 nM with a limit of detection of 2.5 pM Co2 +. The proposed method being highly sensitive, selective, valid and low-cost, may be useful to monitor Mn2 + and Co2 + content in clinical laboratories, food industry and environmental control service.

Rhein Elicits In Vitro Cytotoxicity in Primary Human Liver HL-7702 Cells by Inducing Apoptosis through Mitochondria-Mediated Pathway

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Guy-Armel Bounda

2015-01-01

Full Text Available Objective. To study rhein-induced apoptosis signaling pathway and to investigate its molecular mechanisms in primary human hepatic cells. Results. Cell viability of HL-7702 cells treated with rhein showed significant decrease in dose-dependent manner. Following rhein treatment (25 μM, 50 μM, and 100 μM for 12 h, the detection of apoptotic cells was significantly analyzed by flow cytometry and nuclear morphological changes by Hoechst 33258, respectively. Fatty degeneration studies showed upregulation level of the relevant hepatic markers (P < 0.01. Caspase activities expressed significant upregulation of caspase-3, caspase-9, and caspase-8. Moreover, apoptotic cells by rhein were significantly inhibited by Z-LEHD-FMK and Z-DEVD-FMK, caspase-9 inhibitor, and caspase-3 inhibitor, respectively. Overproduction of reactive oxygen species, lipid peroxidation, and loss of mitochondrial membrane potential were detected by fluorometry. Additionally, NAC, a ROS scavenger, significantly attenuated rhein-induced oxidative damage in HL-7702 cells. Furthermore, real-time qPCR results showed significant upregulation of p53, PUMA, Apaf-1, and Casp-9 and Casp-3 mRNA, with no significant changes of Fas and Cytochrome-c. Immunoblotting revealed significant Cytochrome-c release from mitochondria into cytosol and no change in Fas expression. Conclusion. Taken together, these observations suggested that rhein could induce apoptosis in HL-7702 cells via mitochondria-mediated signal pathway with involvement of oxidative stress mechanism.

Chemodosimeter-based fluorescent detection of L-cysteine after extracted by molecularly imprinted polymers.

Science.gov (United States)

Cai, Xiaoqiang; Li, Jinhua; Zhang, Zhong; Wang, Gang; Song, Xingliang; You, Jinmao; Chen, Lingxin

2014-03-01

A chemodosimeter-based fluorescent detection method coupled with molecularly imprinted polymers (MIPs) extraction was developed for determination of L-cysteine (L-Cys) by combining molecular imprinting technique with fluorescent chemodosimeter. The MIPs prepared by precipitation polymerization with L-Cys as template, possessed high specific surface area of 145 m(2)/g and good thermal stability without decomposition lower than 300 °C, and were successfully applied as an adsorbent with excellent selectivity for L-Cys over other amino acids, and enantioselectivity was also demonstrated. A novel chemodosimeter, rhodamine B1, was synthesized for discriminating L-Cys from its structurally similar homocysteine and glutathione as well as various possibly co-existing biospecies in aqueous solutions with notable fluorescence enhancement when adding L-Cys. As L-Cys was added with increasing concentrations, an emission band peaked at 580 nm occurred and significantly increased in fluorescence intensity, by which the L-Cys could be sensed optically. High detectability up to 12.5 nM was obtained. An excellent linearity was found within the wide range of 0.05-50 μM (r=0.9996), and reasonable relative standard deviations ranging from 0.3% to 3.5% were attained. Such typical features as high selectivity, high sensitivity, easy operation and low cost enabled this MIPs-fluorometry to be potentially applicable for routine detection of trace L-Cys. Copyright © 2013 Elsevier B.V. All rights reserved.

In vitro neuroprotective potential of four medicinal plants against rotenone-induced toxicity in SH-SY5Y neuroblastoma cells.

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Seoposengwe, Keabetswe; van Tonder, Jacob John; Steenkamp, Vanessa

2013-12-12

Lannea schweinfurthii, Zanthoxylum capense, Scadoxus puniceus and Crinum bulbispermum are used traditionally to treat neurological disorders. The aim of this study was to evaluate the cytoprotective potential of the four plants, after induction of toxicity using rotenone, in SH-SY5Y neuroblastoma cells. Cytotoxicity of the plant extracts and rotenone was assessed using the sulforhodamine B (SRB) assay. Fluorometry was used to measure intracellular redox state (reactive oxygen species (ROS) and intracellular glutathione content), mitochondrial membrane potential (MMP) and caspase-3 activity, as a marker of apoptotic cell death. Of the tested plants, the methanol extract of Z. capense was the least cytotoxic; LC50 121.3 ± 6.97 μg/ml, while S. puniceus methanol extract was the most cytotoxic; LC50 20.75 ± 1.47 μg/ml. Rotenone reduced intracellular ROS levels after 24 h exposure. Pre-treating cells with S. puniceus and C. bulbispermum extracts reversed the effects of rotenone on intracellular ROS levels. Rotenone exposure also decreased intracellular glutathione levels, which was counteracted by pre-treatment with any one of the extracts. MMP was reduced by rotenone, which was neutralized by pre-treatment with C. bulbispermum ethyl acetate extract. All extracts inhibited rotenone-induced activation of caspase-3. The studied plants demonstrated anti-apoptotic activity and restored intracellular glutathione content following rotenone treatment, suggesting that they may possess neuroprotective properties.

New amphiphilic glycopolypeptide conjugate capable of self-assembly in water into reduction-sensitive micelles for triggered drug release

Energy Technology Data Exchange (ETDEWEB)

Yang, Hui-Kang [DSAPM Lab and PCFM Lab, Department of Polymer and Materials Science, School of Chemistry and Chemical Engineering, Sun Yat-sen University, Guangzhou 510275 (China); Zhang, Li-Ming, E-mail: ceszhlm@mail.sysu.edu.cn [DSAPM Lab and PCFM Lab, Department of Polymer and Materials Science, School of Chemistry and Chemical Engineering, Sun Yat-sen University, Guangzhou 510275 (China); Guangdong Provincial Key Laboratory of New Drug Design and Evaluation, School of Pharmaceutical Science, Sun Yat-sen University, Guangzhou 510006 (China)

2014-08-01

For the development of biomimetic carriers for stimuli-sensitive delivery of anticancer drugs, a novel amphiphilic glycopolypeptide conjugate containing the disulfide bond was prepared for the first time by the ring-opening polymerization of benzyl glutamate N-carboxy anhydride in the presence of (propargyl carbamate)ethyl dithio ethylamine and then click conjugation with α-azido dextran. Its structure was characterized by Fourier-transform infrared spectroscopy and nuclear magnetic resonance analyses. Owing to its amphiphilic nature, such a conjugate could self assemble into nanosize micelles in aqueous medium, as confirmed by fluorometry, transmission electron microscopy and dynamic light scattering. For the resultant micelles, it was found to encapsulate poorly water-soluble anticancer drug (methotrexate, MTX) with the loading efficiency of 45.2%. By the in vitro drug release tests, the release rate of encapsulated MTX was observed to be accelerated significantly in the presence of 10 mM 1,4-dithio-DL-threitol (DTT), analogous to the intracellular redox potential. - Graphical abstract: New amphiphilic glycopolypeptide conjugate containing the disulfide bond could self-assemble in aqueous solution into reduction-sensitive micelles for triggered release of an anticancer drug (methotrexate, MTX) in the presence of 10 mM 1,4-dithio-DL-threitol (DTT). - Highlights: • Amphiphilic glycopolypeptide conjugate containing disulfide bond was prepared. • Such a conjugate self assembled in aqueous solution into nanosize micelles. • The resultant micelles could encapsulate effectively methotrexate drug. • The drug-loaded micelles showed a reduction-sensitive drug release behavior.

Linking FRRF Derived Photophysiology with Carbon-based Primary Productivity: Insights from Concepts of Cellular Energy Allocation

Science.gov (United States)

Schuback, N.; Schallenberg, C.; Duckham, C.; Flecken, M.; Maldonado, M. T.; Tortell, P. D.

2016-02-01

Active chlorophyll a fluorescence approaches, including fast repetition rate fluorometry (FRRF), have the potential to provide estimates of phytoplankton primary productivity at unprecedented spatial and temporal resolution. FRRF-derived productivity rates are based on estimates of charge separation in photosystem II (ETRRCII), which must be converted into ecologically relevant units of carbon fixation. Understanding sources of variability in the coupling of ETRRCII and carbon fixation provides important physiological insight into phytoplankton photosynthesis, and is critical for the application of FRRF as a primary productivity measurement tool. We present data from a series of experiments during which we simultaneously measured phytoplankton carbon fixation and ETRRCII in the iron-limited NE subarctic Pacific. Our results show significant variability of the derived conversion factor (Ve:C/nPSII), with highest values observed under conditions of excess excitation pressure at the level of photosystem II, caused by high light and/or low iron. Our results will be discussed in the context of metabolic plasticity, which evolved in phytoplankton to simultaneously maximize growth and provide photoprotection under fluctuating light and limiting nutrient availabilities. Because the derived conversion factor is associated with conditions of excess light, it correlates with the expression of non-photochemical quenching (NPQ) in the pigment antenna, also derived from FRRF measurements. Our results demonstrate a significant correlation between NPQ and the conversion factor Ve:C/nPSII, and the potential of this relationship to improve FRRF-based estimates of phytoplankton carbon fixation rates is discussed.

Fluorescence imaging spectroscopy (FIS) for comparing spectra from corn ears naturally and artificially infected with aflatoxin producing fungus.

Science.gov (United States)

Hruska, Zuzana; Yao, Haibo; Kincaid, Russell; Darlington, Dawn; Brown, Robert L; Bhatnagar, Deepak; Cleveland, Thomas E

2013-08-01

In an effort to address the problem of rapid detection of aflatoxin in grain, particularly oilseeds, the current study assessed the spectral differences of aflatoxin production in kernels from a cornfield inoculated with spores from 2 different strains of toxigenic Aspergillus flavus. Aflatoxin production in corn from the same field due to natural infestation was also assessed. A small corn plot in Baton Rouge, La., U.S.A., was used during the 2008-growing season. Two groups of 400 plants were inoculated with 2 different inocula and 1 group of 400 plants was designated as controls. Any contamination detected in the controls was attributed to natural infestation. A subset of each group was imaged with a visible near infra red (VNIR) hyperspectral system under ultra violet (UV) excitation and subsequently analyzed for aflatoxin using affinity column fluorometry. Group differences were statistically analyzed. Results indicate that when all the spectral data across all groups were averaged, any potential differences between groups (treated and untreated) were obscured. However, spectral analysis based on contaminated "hot" pixel classification showed a distinct spectral shift/separation between contaminated and clean ears with fluorescence peaks at 501 and 478 nm, respectively. All inoculated and naturally infected control ears had fluorescence peaks at 501 nm that differed from uninfected corn ears. Results from this study may be useful in evaluating rapid, noninvasive instrumentation and/or methodology for aflatoxin detection in grain. © 2013 Institute of Food Technologists®

Poly-P storage by natural biofilms in streams with varying biogeochemistry

Science.gov (United States)

Carrick, H. J.

2015-12-01

Anthropogenic inputs of nitrogen (N) and phosphorus (P) have increased in many watersheds throughout the world; these inputs have been linked to the eutrophication of inland and coastal waters worldwide. We selected and surveyed 20, third-order streams that supported a range of water column biogeochemical conditions (conductivity, nutrient concentrations) located in the mid-Atlantic region, USA. Biofilm biomass, algal taxonomic composition, and nutrient stoichiometry (C, N, P, and poly-P) were measured at all stream sites. Pulse-amplitude modulation fluorometry (PAM) was used to estimate photosynthetic parameters for stream biofilms (e.g., alpha, Pmax), while microbiology techniques were used to verify poly-P storage by pro- and eukaryotic components of the biofilm (e.g., epi-fluorescent staining). As anticipated, chlorophyll ranged over 2 orders of magnitude among the streams (range 10-1,000 mg/m2). Biofilm chlorophyll and algal biovolume levels increased with water column nutrient contents, while the C:P ratio within the biofilm decreased. Both pro and eukaryotic organisms were present in resident biofilms and actively stored intracellular poly-P. Finally, the rate of photosynthetic within the biofilms appeared to be driven the nutritional condition of the biofilms; pmax and alpha values increased with significantly with stream biofilm poly-P content (r2 = 0.35 and 0.44, respectively). These results indicated that where nutrients are plentiful, biofilms P storage is favored, and this is likely a key regulator of stream biofilm biomass and productivity.

Results of elemental analyses of water and waterborne sediment samples from areas of Alaska proposed for the Chukchi Imuruk National Reserve, Selawik National Wildlife Refuge, and Cape Krusenstern National Monument

International Nuclear Information System (INIS)

Sharp, R.R. Jr.

1978-10-01

During July--August 1976, waters and sediments were collected from streams and lakes over an area of 100,000 km 2 around Kotzebue, Alaska, as part of the National Uranium Resource Evaluation Hydrogeochemical and Stream Sediment Reconnaissance. The work provides multielement results for 949 waters and 886 sediments from 979 locations. Of these, 492 waters and 452 sediments are from 517 locations in the proposed Chukchi Imuruk Reserve; 447 waters and 423 sediments are from 451 locations in the proposed Selawik Wildlife Refuge; and 10 waters and 11 sediments are from 11 locations in the proposed Cape Krusenstern Monument. The field data, with concentrations of 13 elements in the waters and 43 in the sediments, are presented, and the sample locations are shown on accompanying plates. The waters were analyzed for uranium by fluorometry or delayed-neutron counting and calcium, chromium, cobalt, copper, iron, lead, magnesium, manganese, molybdenum, nickel, titanium, and zinc by plasma-source emission spectrography. The sediment samples were analyzed for uranium by delayed-neutron counting, beryllium and lithium by arc-source emission spectrography, bismuth, cadmium, copper, lead, nickel, niobium, silver, tin, and tungsten by x-ray fluorescence, and aluminum, antimony, barium, calcium, cerium, cesium, chlorine, chromium, cobalt, dysprosium, europium, gold, hafnium, iron, lanthanum, lutetium, magnesium, manganese, potassium, rubidium, samarium, scandium, sodium, strontium, tantalum, terbium, thorium, titanium, vanadium, ytterbium, and zinc by neutron activation. Uranium to thorium ratios in each sediment are also provided

Evaluation and comparison of FTA card and CTAB DNA extraction methods for non-agricultural taxa.

Science.gov (United States)

Siegel, Chloe S; Stevenson, Florence O; Zimmer, Elizabeth A

2017-02-01

An efficient, effective DNA extraction method is necessary for comprehensive analysis of plant genomes. This study analyzed the quality of DNA obtained using paper FTA cards prepared directly in the field when compared to the more traditional cetyltrimethylammonium bromide (CTAB)-based extraction methods from silica-dried samples. DNA was extracted using FTA cards according to the manufacturer's protocol. In parallel, CTAB-based extractions were done using the automated AutoGen DNA isolation system. DNA quality for both methods was determined for 15 non-agricultural species collected in situ, by gel separation, spectrophotometry, fluorometry, and successful amplification and sequencing of nuclear and chloroplast gene markers. The FTA card extraction method yielded less concentrated, but also less fragmented samples than the CTAB-based technique. The card-extracted samples provided DNA that could be successfully amplified and sequenced. The FTA cards are also useful because the collected samples do not require refrigeration, extensive laboratory expertise, or as many hazardous chemicals as extractions using the CTAB-based technique. The relative success of the FTA card method in our study suggested that this method could be a valuable tool for studies in plant population genetics and conservation biology that may involve screening of hundreds of individual plants. The FTA cards, like the silica gel samples, do not contain plant material capable of propagation, and therefore do not require permits from the U.S. Department of Agriculture (USDA) Animal and Plant Health Inspection Service (APHIS) for transportation.

Remote monitoring of chlorophyll fluorescence in two reef corals during the 2005 bleaching event at Lee Stocking Island, Bahamas

Science.gov (United States)

Manzello, D.; Warner, M.; Stabenau, E.; Hendee, J.; Lesser, M.; Jankulak, M.

2009-03-01

Zooxanthellae fluorescence was measured in situ, remotely, and in near real-time with a pulse amplitude modulated (PAM) fluorometer for a colony of Siderastrea siderea and Agaricia tenuifolia at Lee Stocking Island, Bahamas during the Caribbean-wide 2005 bleaching event. These colonies displayed evidence of photosystem II (PS II) inactivation coincident with thermal stress and seasonally high doses of solar radiation. Hurricane-associated declines in temperature and light appear to have facilitated the recovery of maximum quantum yield of PS II within these two colonies, although both corals responded differently to individual storms. PAM fluorometry, coupled with long-term measurement of in situ light and temperature, provides much more detail of coral photobiology on a seasonal time scale and during possible bleaching conditions than sporadic, subjective, and qualitative observations. S. siderea displayed evidence of PS II inactivation over a month prior to the issuing of a satellite-based, sea surface temperature (SST) bleaching alert by the National Oceanic and Atmospheric Administration (NOAA). In fact, recovery had already begun in S. siderea when the bleaching alert was issued. Fluorescence data for A. tenuifolia were difficult to interpret because the shaded parts of a colony were monitored and thus did not perfectly coincide with thermal stress and seasonally high doses of solar radiation as in S. siderea. These results further emphasize the limitations of solely monitoring SST (satellite or in situ) as a bleaching indicator without considering the physiological status of coral-zooxanthellae symbioses.

Synergistic effects of diuron and sedimentation on photosynthesis and survival of crustose coralline algae

Energy Technology Data Exchange (ETDEWEB)

Harrington, Lindsay [Australian Institute of Marine Science, Townsville, QLD 4810 (Australia) and James Cook University, Townsville, QLD 4811 (Australia)]. E-mail: l.harrington@aims.gov.au; Fabricius, Katharina [Australian Institute of Marine Science, Townsville, QLD 4810 (Australia)]. E-mail: k.fabricius@aims.gov.au; Eaglesham, Geoff [Queensland Health Scientific Services, 39 Kessels Road, Coopers Plains 4108 (Australia); Negri, Andrew [Australian Institute of Marine Science, Townsville, QLD 4810 (Australia)

2005-07-01

Effects of short-term exposure to sedimentation and diuron, separately and in combination, on the photophysiology and survival of crustose coralline algae (CCA) were examined in controlled time-course experiments, using pulse-amplitude modulation (PAM) chlorophyll fluorometry. These experiments indicated that the effects of sediments and diuron, when applied in isolation, were often reversible, with recovery time dependant upon sediment type and diuron concentration. Exposure to fine (<63 {mu}m grain size), nutrient-rich estuarine sediments reduced effective quantum yields ({delta}F/F {sub m'}) of photosystem II in CCA species more than exposure to the same amount of fine (<63 {mu}m grain size) calcareous sediments. Significant inhibition of photosynthesis ({delta}F/F {sub m'}) was also observed at diuron concentrations 2.9 {mu}g L{sup -1}. Fine estuarine sediments in combination with 0.79 {mu}g L{sup -1} dissolved diuron, caused yields ({delta}F/F {sub m'}) to drop by 60% compared with controls after 24 h. The combined exposure to sediments and diuron also retarded recovery, thus {delta}F/F {sub m'} values were still only 60% of the controls after 9 days recovery in clean seawater. Mortality of CCA was observed in some fragments treated with combinations of sediment and diuron. Our results suggest that sediment deposition and exposure to diuron can negatively affect the photosynthetic activity of CCA, with sedimentation stress being significantly enhanced by the presence of trace concentrations of diuron.

Influence of the variation potential on photosynthetic flows of light energy and electrons in pea.

Science.gov (United States)

Sukhova, Ekaterina; Mudrilov, Maxim; Vodeneev, Vladimir; Sukhov, Vladimir

2018-05-01

Local damage (mainly burning, heating, and mechanical wounding) induces propagation of electrical signals, namely, variation potentials, which are important signals during the life of plants that regulate different physiological processes, including photosynthesis. It is known that the variation potential decreases the rate of CO 2 assimilation by the Calvin-Benson cycle; however, its influence on light reactions has been poorly investigated. The aim of our work was to investigate the influence of the variation potential on the light energy flow that is absorbed, trapped and dissipated per active reaction centre in photosystem II and on the flow of electrons through the chloroplast electron transport chain. We analysed chlorophyll fluorescence in pea leaves using JIP-test and PAM-fluorometry; we also investigated delayed fluorescence. The electrical signals were registered using extracellular electrodes. We showed that the burning-induced variation potential stimulated a nonphotochemical loss of energy in photosystem II under dark conditions. It was also shown that the variation potential gradually increased the flow of light energy absorbed, trapped and dissipated by photosystem II. These changes were likely caused by an increase in the fraction of absorbed light distributed to photosystem II. In addition, the variation potential induced a transient increase in electron flow through the photosynthetic electron transport chain. Some probable mechanisms for the influence of the variation potential on the light reactions of photosynthesis (including the potential role of intracellular pH decrease) are discussed in the work.

Timescales of Growth Response of Microbial Mats to Environmental Change in an Ice-Covered Antarctic Lake

Directory of Open Access Journals (Sweden)

Anne D. Jungblut

2013-01-01

Full Text Available Lake Vanda is a perennially ice-covered, closed-basin lake in the McMurdo Dry Valleys, Antarctica. Laminated photosynthetic microbial mats cover the floor of the lake from below the ice cover to >40 m depth. In recent decades, the water level of Lake Vanda has been rising, creating a “natural experiment” on development of mat communities on newly flooded substrates and the response of deeper mats to declining irradiance. Mats in recently flooded depths accumulate one lamina (~0.3 mm per year and accrue ~0.18 µg chlorophyll-a cm−2 y−1. As they increase in thickness, vertical zonation becomes evident, with the upper 2-4 laminae forming an orange-brown zone, rich in myxoxanthophyll and dominated by intertwined Leptolyngbya trichomes. Below this, up to six phycobilin-rich green/pink-pigmented laminae form a subsurface zone, inhabited by Leptolyngbya, Oscillatoria and Phormidium morphotypes. Laminae continued to increase in thickness for several years after burial, and PAM fluorometry indicated photosynthetic potential in all pigmented laminae. At depths that have been submerged for >40 years, mats showed similar internal zonation and formed complex pinnacle structures that were only beginning to appear in shallower mats. Chlorophyll-a did not change over time and these mats appear to represent resource-limited “climax” communities. Acclimation of microbial mats to changing environmental conditions is a slow process, and our data show how legacy effects of past change persist into the modern community structure.

New amphiphilic glycopolypeptide conjugate capable of self-assembly in water into reduction-sensitive micelles for triggered drug release

International Nuclear Information System (INIS)

Yang, Hui-Kang; Zhang, Li-Ming

2014-01-01

For the development of biomimetic carriers for stimuli-sensitive delivery of anticancer drugs, a novel amphiphilic glycopolypeptide conjugate containing the disulfide bond was prepared for the first time by the ring-opening polymerization of benzyl glutamate N-carboxy anhydride in the presence of (propargyl carbamate)ethyl dithio ethylamine and then click conjugation with α-azido dextran. Its structure was characterized by Fourier-transform infrared spectroscopy and nuclear magnetic resonance analyses. Owing to its amphiphilic nature, such a conjugate could self assemble into nanosize micelles in aqueous medium, as confirmed by fluorometry, transmission electron microscopy and dynamic light scattering. For the resultant micelles, it was found to encapsulate poorly water-soluble anticancer drug (methotrexate, MTX) with the loading efficiency of 45.2%. By the in vitro drug release tests, the release rate of encapsulated MTX was observed to be accelerated significantly in the presence of 10 mM 1,4-dithio-DL-threitol (DTT), analogous to the intracellular redox potential. - Graphical abstract: New amphiphilic glycopolypeptide conjugate containing the disulfide bond could self-assemble in aqueous solution into reduction-sensitive micelles for triggered release of an anticancer drug (methotrexate, MTX) in the presence of 10 mM 1,4-dithio-DL-threitol (DTT). - Highlights: • Amphiphilic glycopolypeptide conjugate containing disulfide bond was prepared. • Such a conjugate self assembled in aqueous solution into nanosize micelles. • The resultant micelles could encapsulate effectively methotrexate drug. • The drug-loaded micelles showed a reduction-sensitive drug release behavior

Swallowable fluorometric capsule for wireless triage of gastrointestinal bleeding.

Science.gov (United States)

Nemiroski, A; Ryou, M; Thompson, C C; Westervelt, R M

2015-12-07

Real-time detection of gastrointestinal bleeding remains a major challenge because there does not yet exist a minimally invasive technology that can both i) monitor for blood from an active hemorrhage and ii) uniquely distinguish it from blood left over from an inactive hemorrhage. Such a device would be an important tool for clinical triage. One promising solution, which we have proposed previously, is to inject a fluorescent dye into the blood stream and to use it as a distinctive marker of active bleeding by monitoring leakage into the gastrointestinal tract with a wireless fluorometer. This paper reports, for the first time to our knowledge, the development of a swallowable, wireless capsule with a built-in fluorometer capable of detecting fluorescein in blood, and intended for monitoring gastrointestinal bleeding in the stomach. The embedded, compact fluorometer uses pinholes to define a microliter sensing volume and to eliminate bulky optical components. The proof-of-concept capsule integrates optics, low-noise analog sensing electronics, a microcontroller, battery, and low power Zigbee radio, all into a cylindrical package measuring 11 mm × 27 mm and weighing 10 g. Bench-top experiments demonstrate wireless fluorometry with a limit-of-detection of 20 nM aqueous fluorescein. This device represents a major step towards a technology that would enable simple, rapid detection of active gastrointestinal bleeding, a capability that would save precious time and resources and, ultimately, reduce complications in patients.

A novel NaV1.5 voltage sensor mutation associated with severe atrial and ventricular arrhythmias.

Science.gov (United States)

Wang, Hong-Gang; Zhu, Wandi; Kanter, Ronald J; Silva, Jonathan R; Honeywell, Christina; Gow, Robert M; Pitt, Geoffrey S

2016-03-01

Inherited autosomal dominant mutations in cardiac sodium channels (NaV1.5) cause various arrhythmias, such as long QT syndrome and Brugada syndrome. Although dozens of mutations throughout the protein have been reported, there are few reported mutations within a voltage sensor S4 transmembrane segment and few that are homozygous. Here we report analysis of a novel lidocaine-sensitive recessive mutation, p.R1309H, in the NaV1.5 DIII/S4 voltage sensor in a patient with a complex arrhythmia syndrome. We expressed the wild type or mutant NaV1.5 heterologously for analysis with the patch-clamp and voltage clamp fluorometry (VCF) techniques. p.R1309H depolarized the voltage-dependence of activation, hyperpolarized the voltage-dependence of inactivation, and slowed recovery from inactivation, thereby reducing the channel availability at physiologic membrane potentials. Additionally, p.R1309H increased the "late" Na(+) current. The location of the mutation in DIIIS4 prompted testing for a gating pore current. We observed an inward current at hyperpolarizing voltages that likely exacerbates the loss-of-function defects at resting membrane potentials. Lidocaine reduced the gating pore current. The p.R1309H homozygous NaV1.5 mutation conferred both gain-of-function and loss-of-function effects on NaV1.5 channel activity. Reduction of a mutation-induced gating pore current by lidocaine suggested a therapeutic mechanism. Copyright © 2016 Elsevier Ltd. All rights reserved.

DNA accumulation on ventilation system filters in university buildings in Singapore.

Science.gov (United States)

Luhung, Irvan; Wu, Yan; Xu, Siyu; Yamamoto, Naomichi; Chang, Victor Wei-Chung; Nazaroff, William W

2017-01-01

Biological particles deposit on air handling system filters as they process air. This study reports and interprets abundance and diversity information regarding biomass accumulation on ordinarily used filters acquired from several locations in a university environment. DNA-based analysis was applied both to quantify (via DNA fluorometry and qPCR) and to characterize (via high-throughput sequencing) the microbial material on filters, which mainly processed recirculated indoor air. Results were interpreted in relation to building occupancy and ventilation system operational parameters. Based on accumulated biomass, average DNA concentrations per AHU filter surface area across nine indoor locations after twelve weeks of filter use were in the respective ranges 1.1 to 41 ng per cm2 for total DNA, 0.02 to 3.3 ng per cm2 for bacterial DNA and 0.2 to 2.0 ng DNA per cm2 for fungal DNA. The most abundant genera detected on the AHU filter samples were Clostridium, Streptophyta, Bacillus, Acinetobacter and Ktedonobacter for bacteria and Aspergillus, Cladosporium, Nigrospora, Rigidoporus and Lentinus for fungi. Conditional indoor airborne DNA concentrations (median (range)) were estimated to be 13 (2.6-107) pg/m3 for total DNA, 0.4 (0.05-8.4) pg/m3 for bacterial DNA and 2.3 (1.0-5.1) pg/m3 for fungal DNA. Conditional airborne concentrations and the relative abundances of selected groups of genera correlate well with occupancy level. Bacterial DNA was found to be more responsive than fungal DNA to differences in occupancy level and indoor environmental conditions.

Uranium hydrogeochemical and stream sediment reconnaissance data release for the Lewistown NTMS Quadrangle, Montana, including concentrations of forty-two additional elements

International Nuclear Information System (INIS)

Shannon, S.S. Jr.

1980-08-01

Totals of 758 water and 1170 sediment samples were collected from 1649 locations in the Levistown quadrangle. Water samples were collected at streams, springs, wells, ponds, and marshes; sediment samples were obtained from streams, springs, and ponds. Histograms and statistical data for uranium concentrations in water and sediment samples and thorium concentrations in sediment samples are given. All samples were collected at the nominal reconnaissance density of one sample location per 10 km 2 . Elemental concentration, field measurement, weather, geologic, and geographic data for each sample location are listed for waters and for sediments. Uranium to thorium (U/Th) ratios for sediment samples are included. Water samples were initially analyzed for uranium by fluorometry. All water samples containing more than 40 ppB U were reanalyzed by delayed-neutron counting. Sediments were analyzed for U and Th as well as Al, Sb, Ba, Be, Bi, Cd, Ca, Ce, Cs, Cl, Cr, Co, Cu, Dy, Eu, Au, Hf, Fe, La, Pb, Li, Lu, Mg, Mn, Ni, Nb, K, Rb, Sa, Sc, Ag, Na, Sr, Ta, Tb, Sn, Ti, W, V, Yb, and Zn. All sediments were analyzed for U by delayed neutron counting. Other elemental concentrations in sediments were determined by neutron activation analysis for 31 elements, by x-ray fluorescence for 9 elements, and by arc-source emission spectrography for 2 elements. Analytical results are reported as parts per million. Descriptions of procedures used for analysis of water and sediments samples as well as analytical precisions and detection limits are given

Confocal fluorometer for diffusion tracking in 3D engineered tissue constructs

Science.gov (United States)

Daly, D.; Zilioli, A.; Tan, N.; Buttenschoen, K.; Chikkanna, B.; Reynolds, J.; Marsden, B.; Hughes, C.

2016-03-01

We present results of the development of a non-contacting instrument, called fScan, based on scanning confocal fluorometry for assessing the diffusion of materials through a tissue matrix. There are many areas in healthcare diagnostics and screening where it is now widely accepted that the need for new quantitative monitoring technologies is a major pinch point in patient diagnostics and in vitro testing. With the increasing need to interpret 3D responses this commonly involves the need to track the diffusion of compounds, pharma-active species and cells through a 3D matrix of tissue. Methods are available but to support the advances that are currently only promised, this monitoring needs to be real-time, non-invasive, and economical. At the moment commercial meters tend to be invasive and usually require a sample of the medium to be removed and processed prior to testing. This methodology clearly has a number of significant disadvantages. fScan combines a fiber based optical arrangement with a compact, free space optical front end that has been integrated so that the sample's diffusion can be measured without interference. This architecture is particularly important due to the "wet" nature of the samples. fScan is designed to measure constructs located within standard well plates and a 2-D motion stage locates the required sample with respect to the measurement system. Results are presented that show how the meter has been used to evaluate movements of samples through collagen constructs in situ without disturbing their kinetic characteristics. These kinetics were little understood prior to these measurements.

Extraction tools for identification of chemical contaminants in estuarine and coastal waters to determine toxic pressure on primary producers.

Science.gov (United States)

Booij, Petra; Sjollema, Sascha B; Leonards, Pim E G; de Voogt, Pim; Stroomberg, Gerard J; Vethaak, A Dick; Lamoree, Marja H

2013-09-01

The extent to which chemical stressors affect primary producers in estuarine and coastal waters is largely unknown. However, given the large number of legacy pollutants and chemicals of emerging concern present in the environment, this is an important and relevant issue that requires further study. The purpose of our study was to extract and identify compounds which are inhibitors of photosystem II activity in microalgae from estuarine and coastal waters. Field sampling was conducted in the Western Scheldt estuary (Hansweert, The Netherlands). We compared four different commonly used extraction methods: passive sampling with silicone rubber sheets, polar organic integrative samplers (POCIS) and spot water sampling using two different solid phase extraction (SPE) cartridges. Toxic effects of extracts prepared from spot water samples and passive samplers were determined in the Pulse Amplitude Modulation (PAM) fluorometry bioassay. With target chemical analysis using LC-MS and GC-MS, a set of PAHs, PCBs and pesticides was determined in field samples. These compound classes are listed as priority substances for the marine environment by the OSPAR convention. In addition, recovery experiments with both SPE cartridges were performed to evaluate the extraction suitability of these methods. Passive sampling using silicone rubber sheets and POCIS can be applied to determine compounds with different structures and polarities for further identification and determination of toxic pressure on primary producers. The added value of SPE lies in its suitability for quantitative analysis; calibration of passive samplers still needs further investigation for quantification of field concentrations of contaminants. Copyright © 2013 Elsevier Ltd. All rights reserved.

Optical properties of the Kara Sea

Science.gov (United States)

Johnson, Donald R.; Asper, Vernon; McClimans, Thomas; Weidemann, Alan

2000-04-01

This study was motivated by the need to understand dispersion processes which affect the redistribution of nuclear wastes in the Arctic from dump sites in the Kara Sea and in the rivers which flow into the Kara Sea. We focus on vertical profiles of light beam transmission and fluorometry made over the delta region fronting the Ob and Yenisey Rivers and over the East Novaya Zemlya Trough (ENZT). The delta region fronting the Ob River Estuary contains a large repository of particles in a dense bottom nepheloid layer with a maximum centered ˜100 km in front of the estuary entrance and covering an area of roughly 200 km diameter. This suspended particle mass repository appears to contain both sediments and detritus and lends credence to the Lisitsyn [1995] concept of the marginal filter zone. In the deep water of the ENZT we found a strong increase of beam attenuation with depth, indicating a relatively large increase of particle mass concentration from ˜50 m to the bottom (depths in excess of 300 m). The strongest concentration was adjacent to the southeast coast of Novaya Zemlya. We suggest that a type of hyperpycnical flow occurs from accumulation of sediments in the bottom waters of Novaya Zemlya fjords which then cascades down the steep slopes adjacent to the island, producing the particle mass distribution as observed by the transmissometer. The accumulation of these repositories of high particle mass concentrations in suspension would suggest that the residence time is high but that storm-driven events could act to disperse the material.

Accumulation and effects of metal mixtures in two seaweed species.

Science.gov (United States)

Jarvis, Tayler A; Bielmyer-Fraser, Gretchen K

2015-05-01

Metal pollution, due to various anthropogenic sources, may pose a threat to marine ecosystems. Metals can be introduced into food chains via bioaccumulation in primary producers, and may potentially lead to toxic effects. Macroalgae are used as food by a wide variety of organisms, and are therefore extremely important in aquatic systems. This study investigated the accumulation and effects of metals in two macroalgae species. The green seaweed, Ulva lactuca and the red seaweed, Agardhiella subulata were each concurrently exposed to five metals (Cu, Ni, Pb, Cd, and Zn) and U. lactuca was also exposed to each metal individually for 48 h. Metal accumulation in the seaweed was measured, and various photosynthetic parameters were assessed, using imaging pulse amplitude modulated (PAM) fluorometry. Increased metal accumulation occurred in both seaweed species after 48 h exposure to metal mixtures and each metal individually. The distribution of metals in both seaweed species changed with increasing metal exposure concentrations, resulting in higher proportions of Cu and Zn in the metal-exposed groups, as compared to respective controls. Further, U. lactuca accumulated higher concentrations of metals when exposed to each metal individually rather than in metal mixtures, suggesting interactions among metals for uptake and/or bioaccumulation. Significant impairment of photosynthetic parameters in U. lactuca was observed after exposure to 100 and 1000 μg/L metal mixtures, as well as 100 μg/L of either Cd or Cu. These results demonstrate metal bioaccumulation and toxic effects in important primary producers, and may have implications for higher trophic levels. Published by Elsevier Inc.

Distribution of C-myc Antisense Oligonucleotides in Rabbits after Local Delivery by Implanted Gelatin Coated Piatinium -iridium Stent

Institute of Scientific and Technical Information of China (English)

张新霞; 庞志功; 崔长琮; 许香广; 胡雪松; 方卫华

2003-01-01

Objectives To assess the feasibility, efficiency and tissue distribution of localdelivered c - myc antisense oligonucleotides (ASODN)by implanted gelatin coated Platinium- Iridium (Pt-Ir) stent. Methods Gelatin coated Pt- Ir stentwhich absorbed carboxyfluorescein - 5 - succimidylester (FAM) labeled c -myc ASODN were implantedin the right carotid arteries of 6 rabbits under vision.Blood samples were collected at the indicated times.The target artery、 left carotid artery、 heart、 liver andkidney obtained at 45 minutes、 2 hours and 6hours. The concentration of c - myc ASODN in plasmaand tissues were determined by Thin Layer Fluorome-try. Tissue distribution of c- myc ASODN were as-sessed by fluorescence microscopy. Results At 45min, 2 h, 6 h, the concentration of FAM labeled c -myc ASODN in target artery was 244.39, 194.44,126.94(μg/g tissues) respectively, and the deliveryefficiency were 44.4% 、 35.4% and 23.1% respec-tively. At the same indicated time point, the plasmaconcentration was 8.41, 5. 83, 14.75 (μg/ml) respec-tively. Therefore c -myc ASODN concentrations in thetarget vessel were 29、 33 and 9 -fold higher than thatin the plasma. There was circumferential distribution oflabeled c -myc in the area of highest fluorescein co-inciding with the site of medial dissecting from stent-ing, and the label was most intense in target vesselmedia harvested at 45 min time point and then dis-persed to adventitia. Conclusions Gelatin coated Pt- Ir stent mediated local delivery of c - myc ASODN isfeasible and efficient. The localization of ASODN ismainly in target vessel wall.

Fluorescence characterization of the natural organic matter in deep ground waters from the Canadian Shield, Ontario, Canada

International Nuclear Information System (INIS)

Francois Caron; Stefan Siemann; Karen Sharp-King; Scott Smith, D.

2010-01-01

Deep groundwater samples from a deep borehole in the Canadian Shield, Ontario, Canada, have been analyzed by fluorometry, to determine the difference in character of the natural organic matter (NOM) with depth. This work was done to obtain a set of geochemical characteristics of deep groundwaters at the site. The fluorescence signal is a complex signature of excitation and emission of light from fluorescent molecules which are part of all natural waters. Fluorescent components have characteristic excitation/emission components, defined as a humic-like (C1), fulvic-like (C2), and protein-like (C3); these are found in various proportions in natural samples. Changes in relative fluorescence intensities of these components have been used in the past to determine the origin and/or processes of the NOM between sampling locations. In this work, six samples were taken at different depths, from ∼108 to 650 m below the surface in the borehole. The fluorescence signals of the samples showed three main patterns: (1) the shallower samples (∼108, 139 and 285 m) had a pattern similar to that of surface groundwaters, dominated by components C1 and C2; (2) the samples in deep groundwaters (∼620 and 650 m) had a weak overall signal, dominated by component C3; finally (3) the mid-depth sample (∼503 m) had a component pattern intermediate between the shallower and deeper zones. This set of data is consistent with other data for the groundwaters from this borehole, such as chlorinity, suggesting that the three sampling intervals represent three different types of groundwaters. (author)

Determination of photosynthetic and enzymatic biomarkers sensitivity used to evaluate toxic effects of copper and fludioxonil in alga Scenedesmus obliquus

Energy Technology Data Exchange (ETDEWEB)

Dewez, David [Departement de Chimie et de Biochimie, Centre TOXEN, Universite du Quebec a Montreal, CP 8888, Succursale Centre-Ville, Montreal, Quebec, H3C 3P8 (Canada); Geoffroy, Laure [Laboratoire d' Eco-Toxicologie, Unite de recherche ' Vignes et Vins de Champagne' , UPRES-EA 2069, Universite de Reims Champagne-Ardenne BP 1039, F51687 REIMS CEDEX 2 (France); Vernet, Guy [Laboratoire d' Eco-Toxicologie, Unite de recherche ' Vignes et Vins de Champagne' , UPRES-EA 2069, Universite de Reims Champagne-Ardenne BP 1039, F51687 REIMS CEDEX 2 (France); Popovic, Radovan [Departement de Chimie et de Biochimie, Centre TOXEN, Universite du Quebec a Montreal, CP 8888, Succursale Centre-Ville, Montreal, Quebec, H3C 3P8 (Canada)]. E-mail: popovic.radovan@uqam.ca

2005-08-30

Modulated PAM fluorometry and Plant Efficiency Analyser methods were used to investigate photosynthetic fluorescence parameters of alga Scenedesmus obliquus exposed to inhibitory effect of fungicides copper sulphate and fludioxonil (N-(4-nitrophenyl)-N'-propyl-uree). The change of those parameters were studied when alga S. obliquus have been exposed during 48 h to different concentrations of fungicides (1, 2 and 3 mg l{sup -1}). Under the same condition, enzymatic activities of catalase, ascorbate peroxidase, glutathione reductase and glutathione S-transferase were investigated to evaluate antioxidative response to fungicides effects. The change of sensitivity of those parameters was dependent to the mode of fungicide action, their concentration and time of exposure. For copper effects, the most indicative photosynthetic biomarkers were parameters Q {sub N} as non-photochemical fluorescence quenching, Q {sub Emax} as the proton induced fluorescence quenching and ABS/RC as the antenna size per photosystem II reaction center. Copper induced oxidative stress was indicated by increased activity of catalase serving as the most sensitive and valuable enzymatic biomarker. On the other hand, fludioxonil effect on photosynthetic parameters was very negligible and consequently not very useful as biomarkers. However, fludioxonil induced strong antioxidative activities associated with cytosol enzymes, as we found for catalase, ascorbate peroxidase and glutathione S-transferase activities. By obtained results, we may suggest for the activation of those enzymes to be sensitive and valuable biomarkers of oxidative stress induced by fludioxonil. Determination of biomarkers sensitivity may offer advantages in providing real criteria to use them for ecotoxicological diagnostic studies.

Potential role of proteasome on c-jun related signaling in hypercholesterolemia induced atherosclerosis

Directory of Open Access Journals (Sweden)

Erdi Sozen

2014-01-01

Full Text Available Atherosclerosis and its complications are major causes of death all over the world. One of the major risks of atherosclerosis is hypercholesterolemia. During atherosclerosis, oxidized low density lipoprotein (oxLDL regulates CD36-mediated activation of c-jun amino terminal kinase-1 (JNK1 and modulates matrix metalloproteinase (MMP induction which stimulates inflammation with an invasion of monocytes. Additionally, inhibition of proteasome leads to an accumulation of c-jun and phosphorylated c-jun and activation of activator protein-1 (AP-1 related increase of MMP expression. We have previously reported a significant increase in cluster of differentiation 36 (CD36 mRNA levels in hypercholesterolemic rabbits and shown that vitamin E treatment prevented the cholesterol induced increase in CD36 mRNA expression. In the present study, our aim is to identify the signaling molecules/transcription factors involved in the progression of atherosclerosis following CD36 activation in an in vivo model of hypercholesterolemic (induced by 2% cholesterol containing diet rabbits. In this direction, proteasomal activities by fluorometry and c-jun, phospo c-jun, JNK1, MMP-9 expressions by quantitative RT-PCR and immunoblotting were tested in aortic tissues. The effects of vitamin E on these changes were also investigated in this model. As a result, c-jun was phosphorylated following decreased proteasomal degradation in hypercholesterolemic group. MMP-9 expression was also increased in cholesterol group rabbits contributing to the development of atherosclerosis. In addition, vitamin E showed its effect by decreasing MMP-9 levels and phosphorylation of c-jun.

Uranium hydrogeochemical and stream sediment reconnaissance data release for the Butte NTMS Quadrangle, Montana, including concentrations of forty-two additional elements

International Nuclear Information System (INIS)

Broxton, D.E.; George, W.E.; Montoya, J.V.; Martell, C.J.; Hensley, W.K.; Hanks, D.

1980-05-01

This report contains data collected during a geochemical survey for uranium in the Butte National Topographic Map Series (NTMS) quadrangle of west-central Montana. Histograms and statistical data for uranium concentrations in water and sediment samples and thorium concentrations in sediment samples are given. Elemental concentration, field measurement, weather, geologic, and geographic data for each sample location are listed for waters and for sediments. Uranium/thorium ratios for sediment samples are also included. This report contains uranium analyses for water samples and multielement analyses for sediment samples. A supplemental report containing the results of multielement analyses of water samples will be open filed in the near future. Sediments were analyzed for uranium and thorium as well as aluminum, antimony, barium, beryllium, bismuth, cadmium, calcium, cerium, cesium, chlorine, chromium, cobalt, copper, dysprosium, europium, gold, hafnium, iron, lanthanum, lead, lithium, lutetium, magnesium, manganese, nickel, niobium, potassium, rubidium, samarium, scandium, silver, sodium, strontium, tantalum, terbium, tin, titanium, tungsten, vanadium, ytterbium, and zinc. All elemental analyses were performed at the LASL. Water samples were initially analyzed for uranium by fluorometry. All water samples containing more than 40 ppB uranium were reanalyzed by delayed-neutron counting (DNC). All sediments were analyzed for uranium by DNC. Other elemental concentrations in sediments were determined by neutron activation analysis for 31 elements, by x-ray fluorescence for 9 elements, and by arc-source emission spectrography for 2 elements. Analytical results for sediments are reported as parts per million. Descriptions of procedures used for analysis of water and sediment samples as well as analytical precisions and detection limits are given

Tissue distribution, metabolism, and residue depletion study in Atlantic salmon following oral administration of [3H]emamectin benzoate.

Science.gov (United States)

Kim-Kang, Heasook; Bova, Alice; Crouch, Louis S; Wislocki, Peter G; Robinson, Robert A; Wu, Jinn

2004-04-07

Atlantic salmon (approximately 1.3 kg) maintained in tanks of seawater at 5 +/- 1 degrees C were dosed with [3H]emamectin B1 benzoate in feed at a nominal rate of 50 microg of emamectin benzoate/kg/day for 7 consecutive days. Tissues, blood, and bile were collected from 10 fish each at 3 and 12 h and at 1, 3, 7, 15, 30, 45, 60, and 90 days post final dose. Feces were collected daily from the tanks beginning just prior to dosing to 90 days post final dose. The total radioactive residues (TRR) of the daily feces samples during dosing were 0.25 ppm maximal, and >97% of the TRR in pooled feces covering the dosing period was emamectin B1a. Feces TRR then rapidly declined to approximately 0.05 ppm by 1 day post final dose. The ranges of mean TRR for tissues over the 90 days post dose period were as follows: kidney, 1.4-3 ppm; liver, 1.0-2.3 ppm; skin, 0.04-0.09 ppm; muscle, 0.02-0.06 ppm; and bone, emamectin B1a (81-100% TRR) and desmethylemamectin B1a (0-17% TRR) with N-formylemamectin B1a seen in trace amounts (emamectin residues was emamectin B1a. The emamectin B1a level was quantified in individual samples of skin and muscle using HPLC-fluorometry and was below 85 ppb in all samples analyzed (3 h to 30 days post dose).

Diurnal variation in the coupling of photosynthetic electron transport and carbon fixation in iron-limited phytoplankton in the NE subarctic Pacific

Science.gov (United States)

Schuback, Nina; Flecken, Mirkko; Maldonado, Maria T.; Tortell, Philippe D.

2016-02-01

Active chlorophyll a fluorescence approaches, including fast repetition rate fluorometry (FRRF), have the potential to provide estimates of phytoplankton primary productivity at an unprecedented spatial and temporal resolution. FRRF-derived productivity rates are based on estimates of charge separation in reaction center II (ETRRCII), which must be converted into ecologically relevant units of carbon fixation. Understanding sources of variability in the coupling of ETRRCII and carbon fixation provides physiological insight into phytoplankton photosynthesis and is critical for the application of FRRF as a primary productivity measurement tool. In the present study, we simultaneously measured phytoplankton carbon fixation and ETRRCII in the iron-limited NE subarctic Pacific over the course of a diurnal cycle. We show that rates of ETRRCII are closely tied to the diurnal cycle in light availability, whereas rates of carbon fixation appear to be influenced by endogenous changes in metabolic energy allocation under iron-limited conditions. Unsynchronized diurnal oscillations of the two rates led to 3.5-fold changes in the conversion factor between ETRRCII and carbon fixation (Kc / nPSII). Consequently, diurnal variability in phytoplankton carbon fixation cannot be adequately captured with FRRF approaches if a constant conversion factor is applied. Utilizing several auxiliary photophysiological measurements, we observed that a high conversion factor is associated with conditions of excess light and correlates with the increased expression of non-photochemical quenching (NPQ) in the pigment antenna, as derived from FRRF measurements. The observed correlation between NPQ and Kc / nPSII requires further validation but has the potential to improve estimates of phytoplankton carbon fixation rates from FRRF measurements alone.

Evaluation and comparison of FTA card and CTAB DNA extraction methods for non-agricultural taxa1

Science.gov (United States)

Siegel, Chloe S.; Stevenson, Florence O.; Zimmer, Elizabeth A.

2017-01-01

Premise of the study: An efficient, effective DNA extraction method is necessary for comprehensive analysis of plant genomes. This study analyzed the quality of DNA obtained using paper FTA cards prepared directly in the field when compared to the more traditional cetyltrimethylammonium bromide (CTAB)–based extraction methods from silica-dried samples. Methods: DNA was extracted using FTA cards according to the manufacturer’s protocol. In parallel, CTAB-based extractions were done using the automated AutoGen DNA isolation system. DNA quality for both methods was determined for 15 non-agricultural species collected in situ, by gel separation, spectrophotometry, fluorometry, and successful amplification and sequencing of nuclear and chloroplast gene markers. Results: The FTA card extraction method yielded less concentrated, but also less fragmented samples than the CTAB-based technique. The card-extracted samples provided DNA that could be successfully amplified and sequenced. The FTA cards are also useful because the collected samples do not require refrigeration, extensive laboratory expertise, or as many hazardous chemicals as extractions using the CTAB-based technique. Discussion: The relative success of the FTA card method in our study suggested that this method could be a valuable tool for studies in plant population genetics and conservation biology that may involve screening of hundreds of individual plants. The FTA cards, like the silica gel samples, do not contain plant material capable of propagation, and therefore do not require permits from the U.S. Department of Agriculture (USDA) Animal and Plant Health Inspection Service (APHIS) for transportation. PMID:28224056

Primary productivity as a control over soil microbial diversity along environmental gradients in a polar desert ecosystem

Directory of Open Access Journals (Sweden)

Kevin M. Geyer

2017-07-01

Full Text Available Primary production is the fundamental source of energy to foodwebs and ecosystems, and is thus an important constraint on soil communities. This coupling is particularly evident in polar terrestrial ecosystems where biological diversity and activity is tightly constrained by edaphic gradients of productivity (e.g., soil moisture, organic carbon availability and geochemical severity (e.g., pH, electrical conductivity. In the McMurdo Dry Valleys of Antarctica, environmental gradients determine numerous properties of soil communities and yet relatively few estimates of gross or net primary productivity (GPP, NPP exist for this region. Here we describe a survey utilizing pulse amplitude modulation (PAM fluorometry to estimate rates of GPP across a broad environmental gradient along with belowground microbial diversity and decomposition. PAM estimates of GPP ranged from an average of 0.27 μmol O2/m2/s in the most arid soils to an average of 6.97 μmol O2/m2/s in the most productive soils, the latter equivalent to 217 g C/m2/y in annual NPP assuming a 60 day growing season. A diversity index of four carbon-acquiring enzyme activities also increased with soil productivity, suggesting that the diversity of organic substrates in mesic environments may be an additional driver of microbial diversity. Overall, soil productivity was a stronger predictor of microbial diversity and enzymatic activity than any estimate of geochemical severity. These results highlight the fundamental role of environmental gradients to control community diversity and the dynamics of ecosystem-scale carbon pools in arid systems.

Patterns in tropical seagrass photosynthesis in relation to light, depth and habitat

Science.gov (United States)

Campbell, Stuart J.; McKenzie, Len J.; Kerville, Simon P.; Bité, Juanita S.

2007-07-01

Seagrass meadows across north-eastern Australia, survive a range of environmental conditions in coastal bays, reefs, estuarine and deepwater habitats through adaptation of a range of structural, morphological and physiological features. The aim of this study was to investigate the influence of spatial features (habitat type, site and depth) and photon flux on the photosynthetic performance of 11 tropical seagrass species. Pulse amplitude modulated (PAM) fluorometry was used to generate rapid light curves from which measures of maximal electron transport rate (ETR max), photosynthetic efficiency ( α), saturating irradiance ( Ek) and effective quantum yield (Δ F/ Fm') were derived. The amount of light absorbed by leaves (absorption factor) was also determined for each population. In intertidal habitats many seagrass species exhibited typical sun-type responses with a close coupling of both ETR max and Ek with photon flux. Photosynthetic performance ranged from minima in Thalassodendron ciliatum to maxima in Syringodium isoetifolium. The absence of a coupling between photosynthetic performance and photon flux in subtidal populations was most likely due to highly variable light climates and possible light attenuation, and hence the photo-biology of estuarine and deepwater seagrasses exhibited photosynthetic responses indicative of light limitation. In contrast seagrass species from shallow reef and coastal habitats for the most part exhibited light saturation characteristics. Of all the variables examined ETR max, Ek and Δ F/ Fm' were most responsive to changing light climates and provide reliable physiological indicators of real-time photosynthetic performance of tropical seagrasses under different light conditions.

Detailed uranium hydrogeochemical and stream sediment reconnaissance data release for the eastern portion of the Montrose NTMS Quadrangle, Colorado, including concentrations of forty-five additional elements

International Nuclear Information System (INIS)

Maassen, L.W.

1981-01-01

In September and October 1979, the Los Alamos Scientific Laboratory (LASL) conducted a detailed geochemical survey for uranium primarily in the Sawatch Range in the eastern part of the Montrose National Topographic Map Series (NTMS) quadrangle, Colorado, as part of the National Hydrogeochemical and Stream Sediment Reconnaissance (HSSR). Totals of 1034 water and 2087 sediment samples were collected from streams and springs from 2088 locations within a 5420-km 2 area. Statistical data for uranium concentrations in water and sediment samples are presented. Elemental concentration, field measurement, weather, geologic, and geographic data for each sample location are listed for waters and for sediments in appendices. Uranium/thorium ratios for sediment samples are also included. This report contains uranium analyses for water samples and multielement analyses for sediment samples. Sediments were analyzed for uranium and thorium as well as Al, Sb, As, Ba, Be, Bi, Cd, Ca, Ce, Cs, Cl, Cr, Co, Cu, Dy, Eu, Au, Hf, Fe, La, Pb, Li, Lu, Mg, Mn, Ni, Nb, K, Rb, Sm, Sc, Se, Ag, Na, Sr, Ta, Tb, Sn, Ti, W, V, Yb, Zn, and Zr. All elemental analyses were performed at the LASL. Water samples were analyzed for uranium by fluorometry. Sediments were analyzed for uranium by delayed neutron counting. Other elemental concentrations in sediments were determined by neutron activation analysis for 31 elements, by x-ray fluorescence for 12 elements, and by arc-source emission spectrography for 2 elements. Descriptions of procedures as analytical precisions and detection limits are given in the appendix

Determination of photosynthetic and enzymatic biomarkers sensitivity used to evaluate toxic effects of copper and fludioxonil in alga Scenedesmus obliquus

International Nuclear Information System (INIS)

Dewez, David; Geoffroy, Laure; Vernet, Guy; Popovic, Radovan

2005-01-01

Modulated PAM fluorometry and Plant Efficiency Analyser methods were used to investigate photosynthetic fluorescence parameters of alga Scenedesmus obliquus exposed to inhibitory effect of fungicides copper sulphate and fludioxonil (N-(4-nitrophenyl)-N'-propyl-uree). The change of those parameters were studied when alga S. obliquus have been exposed during 48 h to different concentrations of fungicides (1, 2 and 3 mg l -1 ). Under the same condition, enzymatic activities of catalase, ascorbate peroxidase, glutathione reductase and glutathione S-transferase were investigated to evaluate antioxidative response to fungicides effects. The change of sensitivity of those parameters was dependent to the mode of fungicide action, their concentration and time of exposure. For copper effects, the most indicative photosynthetic biomarkers were parameters Q N as non-photochemical fluorescence quenching, Q Emax as the proton induced fluorescence quenching and ABS/RC as the antenna size per photosystem II reaction center. Copper induced oxidative stress was indicated by increased activity of catalase serving as the most sensitive and valuable enzymatic biomarker. On the other hand, fludioxonil effect on photosynthetic parameters was very negligible and consequently not very useful as biomarkers. However, fludioxonil induced strong antioxidative activities associated with cytosol enzymes, as we found for catalase, ascorbate peroxidase and glutathione S-transferase activities. By obtained results, we may suggest for the activation of those enzymes to be sensitive and valuable biomarkers of oxidative stress induced by fludioxonil. Determination of biomarkers sensitivity may offer advantages in providing real criteria to use them for ecotoxicological diagnostic studies

Uptake, delivery, and anticancer activity of thymoquinone nanoparticles in breast cancer cells

Energy Technology Data Exchange (ETDEWEB)

Fakhoury, Isabelle [American University of Beirut, Department of Biology (Lebanon); Saad, Walid [American University of Beirut, Department of Chemical and Petroleum Engineering (Lebanon); Bouhadir, Kamal [American University of Beirut, Department of Chemistry (Lebanon); Nygren, Peter [Uppsala University, Department of Immunology, Genetics and Pathology, Experimental and Clinical Oncology (Sweden); Schneider-Stock, Regine [University of Erlangen-Nuremberg, Experimental Tumor Pathology, Institute for Pathology (Germany); Gali-Muhtasib, Hala, E-mail: amro@aub.edu.lb [American University of Beirut, Department of Biology (Lebanon)

2016-07-15

Thymoquinone (TQ) is a promising anticancer molecule but its development is hindered by its limited bioavailability. Drug encapsulation is commonly used to overcome low drug solubility, limited bioavailability, and nonspecific targeting. In this project, TQ nanoparticles (TQ-NP) were synthesized and characterized. The cytotoxicity of the NP was investigated in nontumorigenic MCF-10-A breast cells, while the uptake, distribution, as well as the anticancer potential were investigated in MCF-7 and MDA-MB-231 breast cancer cells. Flash Nanoprecipitation and dynamic light scattering coupled with scanning electron microscopy were used to prepare and characterize TQ-NP prior to measuring their anticancer potential by MTT assay. The uptake and subcellular intake of TQ-NP were evaluated by fluorometry and confocal microscopy. TQ-NP were stable with a hydrodynamic average diameter size around 100 nm. Entrapment efficiency and loading content of TQ-NP were high (around 80 and 50 %, respectively). In vitro, TQ-NP had equal or enhanced anticancer activity effects compared to TQ in MCF-7 and aggressive MDA-MB-231 breast cancer cells, respectively, with no significant cytotoxicity of the blank NP. In addition, TQ and TQ-NP were relatively nontoxic to MCF-10-A normal breast cells. TQ-NP uptake mechanism was both time and concentration dependent. Treatment with inhibitors of endocytosis suggested the involvement of caveolin in TQ-NP uptake. This was further confirmed by subcellular localization findings showing the colocalization of TQ-NP with caveolin and transferrin as well as with the early and late markers of endocytosis. Altogether, the results describe an approach for the enhancement of TQ anticancer activity and uncover the mechanisms behind cell-TQ-NP interaction.Graphical Abstract.

Los Alamos Scientific Laboratory approach to hydrogeochemical and stream sediment reconnaissance for uranium in the United States

International Nuclear Information System (INIS)

Bolivar, S.L.

1981-01-01

The Los Alamos Scientific Laboratory of the United States is conducting a geochemical survey for uranium in the Rocky Mountain states of New Mexico, Colorado, Wyoming, and Montana and in Alaska. This survey is part of a national hydrogeochemical and stream sediment reconnaissance in which four Department of Energy laboratories will study the uranium resources of the United States to provide data for the National Uranium Resource Evaluation program. The reconnaissance will identify areas having higher than background concentrations of uranium in ground waters, surface waters, and water-transported sediments. Water and sediment samples are collected at a nominal density of one sample location per 10 km 2 except for lake areas of Alaska where the density is one sample location per 23 km 2 . Water samples are analyzed for uranium by fluorometry which has a 0.02 parts per billion lower limit of detection. Concentrations of 12 additional elements in water are determined by plasma-source emission spectrography. All sediments are analyzed for uranium by delayed-neutron counting with a 20 parts per billion lower limit of detection, which is well below the range of uranium concentrations in natural sediment samples. Elemental concentrations in sediments are also determined by neutron activation analysis for 31 elements by x-ray fluorescence for 9 elements, and by arc-source emission spectrography for 2 elements. The multielement analyses provide valuable data for studies concerning pathfinder elements, environmental pollution, elemental distributions, dispersion halos, and economic ore deposits other than uranium. To date, all of four Rocky Mountain states and about 80% of Alaska have been sampled. About 220,000 samples have been collected from an area of nearly 2,500,000 km 2

Hypoxia-targeting antitumor prodrugs and photosensitizers

International Nuclear Information System (INIS)

Zhang Zhouen; Nishimoto, S.I.

2006-01-01

Tumor hypoxia has been identified as a key subject for tumor therapy, since hypoxic tumor cells show resistance to treatment of tumor tissues by radiotherapy, chemotherapy and phototherapy. For improvement of tumor radiotherapy, we have proposed a series of radiation-activated prodrugs that could selectively release antitumor agent 5-fluorouracil or 5-fluorodeoxyuridine under hypoxic conditions. Recently, we attempted to develop two families of novel hypoxia-targeting antitumor agents, considering that tumor-hypoxic environment is favorable to biological and photochemical reductions. The first family of prodrugs was derived from camptothecin as a potent topoisomerase I inhibitor and several bioreductive motifs. These prodrugs could be activated by NADPH-cytochrome P450 reductase or DT-diaphorase to release free camptothecin, and thereby showed hypoxia-selective cytotoxictiy towards tumor cells. These prodrugs were also applicable to the real-time monitoring of activation and antitumor effect by fluorometry. Furthermore, the camptothecin-bioreductive motif conjugates was confirmed to show an oxygen-independent DAN photocleaving activity, which could overcome a drawback of back electron transfer occurring in the photosensitized one-electron oxidation of DNA. Thus, these camptothecin derivatives could be useful to both chemotherapy and phototherapy for hypoxic tumor cells. The second family of prodrugs harnessed UV light for cancer therapy, incorporating the antitumor agent 5-fluorourcil and the photolabile 2-nitrobenzyl chromophores. The attachment of a tumor-homing cyclic peptide CNGRC was also employed to construct the prototype of tumor-targeting photoactiaved antitumor prodrug. These novel prodrugs released high yield of 5-fluorourcil upon UV irradiation at λ ex =365 nm, while being quite stable in the dark. The photoactivation mechanism was also clarified by means of nanosecond laser flash photolysis. (authors)

Targeted Modification of Mitochondrial ROS Production Converts High Glucose-Induced Cytotoxicity to Cytoprotection: Effects on Anesthetic Preconditioning.

Science.gov (United States)

Sedlic, Filip; Muravyeva, Maria Y; Sepac, Ana; Sedlic, Marija; Williams, Anna Marie; Yang, Meiying; Bai, Xiaowen; Bosnjak, Zeljko J

2017-01-01

Contradictory reports on the effects of diabetes and hyperglycemia on myocardial infarction range from cytotoxicity to cytoprotection. The study was designed to investigate acute effects of high glucose-driven changes in mitochondrial metabolism and osmolarity on adaptive mechanisms and resistance to oxidative stress of isolated rat cardiomyocytes. We examined the effects of high glucose on several parameters of mitochondrial bioenergetics, including changes in oxygen consumption, mitochondrial membrane potential, and NAD(P)H fluorometry. Effects of high glucose on the endogenous cytoprotective mechanisms elicited by anesthetic preconditioning (APC) and the mediators of cell injury were also tested. These experiments included real-time measurements of reactive oxygen species (ROS) production and mitochondrial permeability transition pore (mPTP) opening in single cells by laser scanning fluorescence confocal microscopy, and cell survival assay. High glucose rapidly enhanced mitochondrial energy metabolism, observed by increase in NAD(P)H fluorescence intensity, oxygen consumption, and mitochondrial membrane potential. This substantially elevated production of ROS, accelerated opening of the mPTP, and decreased survival of cells exposed to oxidative stress. Abrogation of high glucose-induced mitochondrial hyperpolarization with 2,4 dinitrophenol (DNP) significantly, but not completely, attenuated ROS production to a level similar to hyperosmotic mannitol control. DNP treatment reversed high glucose-induced cytotoxicity to cytoprotection. Hyperosmotic mannitol treatment also induced cytoprotection. High glucose abrogated APC-induced mitochondrial depolarization, delay in mPTP opening and cytoprotection. In conclusion, high glucose-induced mitochondrial hyperpolarization abolishes APC and augments cell injury. Attenuation of high glucose-induced ROS production by eliminating mitochondrial hyperpolarization protects cardiomyocytes. J. Cell. Physiol. 232: 216-224, 2017

Uranium Hydrogeochemical and Stream Sediment Reconnaissance in southwestern Montana

Energy Technology Data Exchange (ETDEWEB)

Broxton, D.E.

1978-02-01

The Los Alamos Scientific Laboratory conducted a Hydrogeochemical and Stream Sediment Reconnaissance in southwestern Montana from early August to mid-October of 1976. A total of 1240 water and 1933 sediment samples were collected from 1994 locations at a nominal density of one location per 10 km/sup 2/. The water samples were collected from streams, wells, and springs; sediment samples were taken at streams and springs. All samples were analyzed at Los Alamos for total uranium by fluorometry or delayed-neutron counting. The uranium content of water samples ranges from below the detection limit (less than 0.3 ppB) to 45.30 ppB and has a mean value of 1.40 ppB. The uranium content of the sediment samples ranges between 0.20 and 206.80 ppM and averages 6.12 ppM. The chosen uranium anomaly threshold value was 7 ppB for surface waters (streams), 9 ppB for groundwaters (wells and springs), and 25 ppM for all sediment samples. The study area consists of the following lithologic groups: Precambrian basement complex, Precambrian Belt metasediments, Paleozoic and Mesozoic shelf sediments, Cretaceous and early Tertiary volcanic and plutonic rocks, Laramide orogenic clastic sediments, and middle to late Tertiary volcanic rocks and intermontane basin sediments. Most of the anomalous water and sediment samples with well-developed dispersion trains occur in areas underlain by or adjacent to silicic plutonic rocks of the Idaho and Boulder batholiths. These anomalies may indicate the presence of uraniferous veins and pegmatites similar to those already known to exist in the area. Fewer anomalous water samples occur in areas underlain by Precambrian basement complex and Tertiary basin fill.

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

Science.gov (United States)

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.

Algae viability over time in a ballast water sample

Science.gov (United States)

Gollasch, Stephan; David, Matej

2018-03-01

The biology of vessels' ballast water needs to be analysed for several reasons, one of these being performance tests of ballast water management systems. This analysis includes a viability assessment of phytoplankton. To overcome logistical problems to get algae sample processing gear on board of a vessel to document algae viability, samples may be transported to land-based laboratories. Concerns were raised how the storage conditions of the sample may impact algae viability over time and what the most appropriate storage conditions were. Here we answer these questions with a long-term algae viability study with daily sample analysis using Pulse-Amplitude Modulated (PAM) fluorometry. The sample was analysed over 79 days. We tested different storage conditions: fridge and room temperature with and without light. It seems that during the first two weeks of the experiment the viability remains almost unchanged with a slight downwards trend. In the continuing period, before the sample was split, a slightly stronger downwards viability trend was observed, which occurred at a similar rate towards the end of the experiment. After the sample was split, the strongest viability reduction was measured for the sample stored without light at room temperature. We concluded that the storage conditions, especially regarding temperature and light exposure, have a stronger impact on algae viability compared to the storage duration and that inappropriate storage conditions reduce algal viability. A sample storage time of up to two weeks in a dark and cool environment has little influence on the organism viability. This indicates that a two week time duration between sample taking on board a vessel and the viability measurement in a land-based laboratory may not be very critical.

Uranium Hydrogeochemical and Stream Sediment Reconnaissance in southwestern Montana

International Nuclear Information System (INIS)

Broxton, D.E.

1978-02-01

The Los Alamos Scientific Laboratory conducted a Hydrogeochemical and Stream Sediment Reconnaissance in southwestern Montana from early August to mid-October of 1976. A total of 1240 water and 1933 sediment samples were collected from 1994 locations at a nominal density of one location per 10 km 2 . The water samples were collected from streams, wells, and springs; sediment samples were taken at streams and springs. All samples were analyzed at Los Alamos for total uranium by fluorometry or delayed-neutron counting. The uranium content of water samples ranges from below the detection limit (less than 0.3 ppB) to 45.30 ppB and has a mean value of 1.40 ppB. The uranium content of the sediment samples ranges between 0.20 and 206.80 ppM and averages 6.12 ppM. The chosen uranium anomaly threshold value was 7 ppB for surface waters (streams), 9 ppB for groundwaters (wells and springs), and 25 ppM for all sediment samples. The study area consists of the following lithologic groups: Precambrian basement complex, Precambrian Belt metasediments, Paleozoic and Mesozoic shelf sediments, Cretaceous and early Tertiary volcanic and plutonic rocks, Laramide orogenic clastic sediments, and middle to late Tertiary volcanic rocks and intermontane basin sediments. Most of the anomalous water and sediment samples with well-developed dispersion trains occur in areas underlain by or adjacent to silicic plutonic rocks of the Idaho and Boulder batholiths. These anomalies may indicate the presence of uraniferous veins and pegmatites similar to those already known to exist in the area. Fewer anomalous water samples occur in areas underlain by Precambrian basement complex and Tertiary basin fill

Lanthanide ions induce hydrolysis of hemoglobin-bound 2,3-diphosphoglycerate (2,3-DPG), conformational changes of globin and bidirectional changes of 2,3-DPG-hemoglobin's oxygen affinity.

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Cheng, Y; Lin, H; Xue, D; Li, R; Wang, K

2001-02-14

The changes in structure and function of 2,3-diphosphoglycerate-hemoglobin (2,3-DPG-Hb) induced by Ln(3+) binding were studied by spectroscopic methods. The binding of lanthanide cations to 2,3-DPG is prior to that to Hb. Ln(3+) binding causes the hydrolysis of either one from the two phosphomonoester bonds in 2,3-DPG non-specifically. The results using the ultrafiltration method indicate that Ln(3+) binding sites for Hb can be classified into three categories: i.e. positive cooperative sites (N(I)), non-cooperative strong sites (N(S)) and non-cooperative weak sites (N(W)) with binding constants in decreasing order: K(I)>K(S)>K(W). The total number of binding sites amounts to about 65 per Hb tetramer. Information on reaction kinetics was obtained from the change of intrinsic fluorescence in Hb monitored by stopped-flow fluorometry. Fluctuation of fluorescence dependent on Ln(3+) concentration and temperature was observed and can be attributed to the successive conformational changes induced by Ln(3+) binding. The results also reveal the bidirectional changes of the oxygen affinity of Hb in the dependence on Ln(3+) concentration. At the range of [Ln(3+)]/[Hb]<2, the marked increase of oxygen affinity (P(50) decrease) with the Ln(3+) concentration can be attributed to the hydrolysis of 2,3-DPG, while the slight rebound of oxygen affinity in higher Ln(3+) concentration can be interpreted by the transition to the T-state of the Hb tetramer induced by Ln(3+) binding. This was indicated by the changes in secondary structure characterized by the decrease of alpha-helix content.

Irradiance optimization of outdoor microalgal cultures using solar tracked photobioreactors.

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Hindersin, Stefan; Leupold, Marco; Kerner, Martin; Hanelt, Dieter

2013-03-01

Photosynthetic activity and temperature regulation of microalgal cultures (Chlorella vulgaris and Scenedesmus obliquus) under different irradiances controlled by a solar tracker and different cell densities were studied in outdoor flat panel photobioreactors. An automated process control unit regulated light and temperature as well as pH value and nutrient concentration in the culture medium. CO2 was supplied using flue gas from an attached combined block heat and power station. Photosynthetic activity was determined by pulse amplitude modulation fluorometry. Compared to the horizontal irradiance of 55 mol photons m(-2) d(-1) on a clear day, the solar tracked photobioreactors enabled a decrease and increase in the overall light absorption from 19 mol photons m(-2) d(-1) (by rotation out of direct irradiance) to 79 mol photons m(-2) d(-1) (following the position of the sun). At biomass concentrations below 1.1 g cell dry weight (CDW) L(-1), photoinhibition of about 35 % occurred at irradiances of ≥1,000 μmol photons m(-2) s(-1) photosynthetic active radiation (PAR). Using solar tracked photobioreactors, photoinhibition can be reduced and at optimum biomass concentration (≥2.3 g CDW L(-1)), the culture was irradiated up to 2,000 μmol photons m(-2) s(-1) to overcome light limitation with biomass yields of 0.7 g CDW mol photons(-1) and high photosynthetic activities indicated by an effective quantum yield of 0.68 and a maximum quantum yield of 0.80 (F v/F m). Overheating due to high irradiance was avoided by turning the PBR out of the sun or using a cooling system, which maintained the temperature close to the species-specific temperature optima.

Los Alamos Scientific Laboratory approach to hydrogeochemical and stream sediment reconnaissance for uranium in the United States

International Nuclear Information System (INIS)

Bolivar, S.L.

1980-01-01

The Los Alamos Scientific Laboratory of the United States is conducting a geochemical survey for uranium in the Rocky Mountain states of New Mexico, Colorado, Wyoming, and Montana and in Alaska. This survey is part of a national hydrogeochemical and stream sediment reconnaissance in which four Department of Energy laboratories will study the uranium resources of the United States to provide data for the National Uranium Resource Evaluation program. The reconnaissance will identify areas having higher than background concentrations of uranium in ground waters, surface waters, and water-transported sediments. The reconnaissance data will be combined with data from airborne radiometric surveys and geological and geophysical investigations to provide an improved estimate for the economics and availability of nuclear fuel resources in the United States and to make information available to industry for use in the exploration and development of uranium resources. Water samples are analyzed for uranium by fluorometry which has a 0.02 parts per billion lower limit of detection. Concentrations of 12 additional elements in water are determined by plasma-source emission spectrography. All sediments are analyzed for uranium by delayed-neutron counting and a 20 parts per billion lower limit of detection. Elemental concentrations in sediments are also determined by neutron activation analysis, x-ray fluorescence, and by arc-source emission spectrography. To date, all of four Rocky Mountain states and about 80% of Alaska have been sampled. About 220,000 samples have been collected from an area of nearly 2,500,000 km 2 . The philosophy, sampling methodology, analytical techniques, and progress of the reconnaissance are described in several published pilot study, reconnaissance, and technical reports. The Los Alamos program was designed to maximize the identification of uranium in terrains of varied geography, geology, and climate

Effects of different algaecides on the photosynthetic capacity, cell integrity and microcystin-LR release of Microcystis aeruginosa

International Nuclear Information System (INIS)

Zhou, Shiqing; Shao, Yisheng; Gao, Naiyun; Deng, Yang; Qiao, Junlian; Ou, Huase; Deng, Jing

2013-01-01

Bench scale tests were conducted to study the effects of four common algaecides, including copper sulfate, hydrogen peroxide, diuron and ethyl 2-methylacetoacetate (EMA) on the photosynthetic capacity, cell integrity and microcystin-LR (MC-LR) release of Microcystis aeruginosa. The release of potassium (K + ) from cell membrane during algaecide exposure was also analyzed. The three typical photosynthetic parameters, including the effective quantum yield (φ e ), photosynthetic efficiency (α) and maximal electron transport rate (rETR max ), were measured by a pulse amplitude modulated (PAM) fluorometry. Results showed that the photosynthetic capacity was all inhibited by the four algaecides, to different degrees, by limiting the energy capture in photosynthesis, and blocking the electron transfer chain in primary reaction. For example, at high diuron concentration (7.5 mg L −1 ), φ e , α and rETR max decreased from 0.46 to 0.19 (p −2 s −1 /μmol photons m −2 s −1 , and from 160.7 to 0.1 (p −2 s −1 compared with the control group after 96 h of exposure, respectively. Furthermore, the increase of algaecide dose could lead to the cell lysis, as well as release of intracellular MC-LR that enhanced the accumulation of extracellular MC-LR. The order of MC-LR release potential for the four algaecides was CuSO 4 > H 2 O 2 > diuron > EMA. Highlights: • PAM was used to investigate the effects of algaecides on Microcystis aeruginosa. • We estimate the release of potassium (K + ) from cell membrane for cell lysis. • The risk of microcystin-LR release was evaluated after algaecides exposure. • The order of MC-LR release potential was copper sulfate > hydrogen peroxide > diuron > ethyl 2-methylacetoacetate

NETosis before and after Hyperglycemic Control in Type 2 Diabetes Mellitus Patients.

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Agostina Carestia

Full Text Available Diabetes is characterized by chronic inflammation, endothelial dysfunction, increased risk of infections and early cardiovascular disease. By releasing neutrophil extracellular traps (NETs, neutrophils kill bacteria and exert pro-inflammatory and pro-thrombotic activities. Increased NETosis has been found in cross-sectional studies including treated type 2 diabetes mellitus (T2DM patients. In this study, we determined whether the ability of neutrophils to form NETs differs in diabetic patients pre- and post-hyperglycemic control versus healthy donors (HD, and the relationship between NETosis with pro-thrombotic, pro-inflammatory biomarkers and thrombotic clinical events.Diabetic patients recently diagnosed and after 6 and 12 months of treatment (N = 25 and HD (N = 25 were included. NET formation was studied by microscopy and fluorometry. Nucleosomes, HNE-DNA complexes, von Willebrand factor (vWF, IL6 and TNFα plasma levels were measured by ELISA and P-selectin on the platelet surface was assessed by cytometry.Basal levels of NETs in recently diagnosed T2DM patients were higher compared to HD. While TNFα stimulation of control neutrophils resulted in DNA release, patient neutrophils were not responsive. Although glycemia decreased after 6 months of metformin treatment, basal and TNFα and PMA-stimulated NETs reached normal values after 12 months. Compared to controls, nucleosomes, HNE-DNA complexes, IL-6 and TNFα levels were increased in recently diagnosed patients and decreased after 12 months of treatment. P-selectin and vWF levels were similar in both populations.Our data suggest that NETs could represent a biomarker for T2DM. Increased NETosis in T2DM patients does not appear to be the consequence of impaired glycemic control but rather due to pro-inflammatory cytokines and is not related to thrombotic events.

Hemodynamic, morphometric and autonomic patterns in hypertensive rats - renin-angiotensin system modulation

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Fernanda S. Zamo

2010-01-01

Full Text Available BACKGROUND: Spontaneously hypertensive rats develop left ventricular hypertrophy, increased blood pressure and blood pressure variability, which are important determinants of heart damage, like the activation of renin-angiotensin system. AIMS: To investigate the effects of the time-course of hypertension over 1 hemodynamic and autonomic patterns (blood pressure; blood pressure variability; heart rate; 2 left ventricular hypertrophy; and 3 local and systemic Renin-angiotensin system of the spontaneously hypertensive rats. METHODS: Male spontaneously hypertensive rats were randomized into two groups: young (n=13 and adult (n=12. Hemodynamic signals (blood pressure, heart rate, blood pressure variability (BPV and spectral analysis of the autonomic components of blood pressure were analyzed. LEFT ventricular hypertrophy was measured by the ratio of LV mass to body weight (mg/g, by myocyte diameter (μm and by relative fibrosis area (RFA, %. ACE and ACE2 activities were measured by fluorometry (UF/min, and plasma renin activity (PRA was assessed by a radioimmunoassay (ng/mL/h. Cardiac gene expressions of Agt, Ace and Ace2 were quantified by RT-PCR (AU. RESULTS: The time-course of hypertension in spontaneously hypertensive rats increased BPV and reduced the alpha index in adult spontaneously hypertensive rats. Adult rats showed increases in left ventricular hypertrophy and in RFA. Compared to young spontaneously hypertensive rats, adult spontaneously hypertensive rats had lower cardiac ACE and ACE2 activities, and high levels of PRA. No change was observed in gene expression of Renin-angiotensin system components. CONCLUSIONS: The observed autonomic dysfunction and modulation of Renin-angiotensin system activity are contributing factors to end-organ damage in hypertension and could be interacting. Our findings suggest that the management of hypertensive disease must start before blood pressure reaches the highest stable levels and the consequent

Los Alamos Scientific Laboratory approach to hydrogeochemical and stream sediment reconnaissance for uranium in the United States

Energy Technology Data Exchange (ETDEWEB)

Bolivar, S.L.

1981-01-01

The Los Alamos Scientific Laboratory of the United States is conducting a geochemical survey for uranium in the Rocky Mountain states of New Mexico, Colorado, Wyoming, and Montana and in Alaska. This survey is part of a national hydrogeochemical and stream sediment reconnaissance in which four Department of Energy laboratories will study the uranium resources of the United States to provide data for the National Uranium Resource Evaluation program. The reconnaissance will identify areas having higher than background concentrations of uranium in ground waters, surface waters, and water-transported sediments. Water and sediment samples are collected at a nominal density of one sample location per 10 km/sup 2/ except for lake areas of Alaska where the density is one sample location per 23 km/sup 2/. Water samples are analyzed for uranium by fluorometry which has a 0.02 parts per billion lower limit of detection. Concentrations of 12 additional elements in water are determined by plasma-source emission spectrography. All sediments are analyzed for uranium by delayed-neutron counting with a 20 parts per billion lower limit of detection, which is well below the range of uranium concentrations in natural sediment samples. Elemental concentrations in sediments are also determined by neutron activation analysis for 31 elements by x-ray fluorescence for 9 elements, and by arc-source emission spectrography for 2 elements. The multielement analyses provide valuable data for studies concerning pathfinder elements, environmental pollution, elemental distributions, dispersion halos, and economic ore deposits other than uranium. To date, all of four Rocky Mountain states and about 80% of Alaska have been sampled. About 220,000 samples have been collected from an area of nearly 2,500,000 km/sup 2/.

Novel Mechanism of Attenuation of LPS-Induced NF-κB Activation by the Heat Shock Protein 90 Inhibitor, 17-N-allylamino-17-demethoxygeldanamycin, in Human Lung Microvascular Endothelial Cells

Science.gov (United States)

Thangjam, Gagan S.; Dimitropoulou, Chistiana; Joshi, Atul D.; Barabutis, Nektarios; Shaw, Mary C.; Kovalenkov, Yevgeniy; Wallace, Chistopher M.; Fulton, David J.; Patel, Vijay

2014-01-01

Heat shock protein (hsp) 90 inhibition attenuates NF-κB activation and blocks inflammation. However, the precise mechanism of NF-κB regulation by hsp90 in the endothelium is not clear. We investigated the mechanisms of hsp90 inhibition by 17-N-allylamino-17-demethoxygeldanamycin (17-AAG) on NF-κB activation by LPS in primary human lung microvascular endothelial cells. Transcriptional activation of NF-κB was measured by luciferase reporter assay, gene expression by real-time RT-PCR, DNA binding of transcription factors by chromatin immunoprecipitation assay, protein–protein interaction by coimmunoprecipitation/immunoblotting, histone deacetylase (HDAC)/histone acetyltransferase enzyme activity by fluorometry, and nucleosome eviction by partial microccocal DNase digestion. In human lung microvascular endothelial cells, 17-AAG–induced degradation of IKBα was accomplished regardless of the phosphorylation/ubiquitination state of the protein. Hence, 17-AAG did not block LPS-induced NF-κB nuclear translocation and DNA binding activity. Instead, 17-AAG blocked the recruitment of the coactivator, cAMP response element binding protein binding protein, and prevented the assembly of a transcriptionally competent RNA polymerase II complex at the κB elements of the IKBα (an NF-κB–responsive gene) promoter. The effect of LPS on IKBα mRNA expression was associated with rapid deacetylation of histone-H3(Lys9) and a dramatic down-regulation of core histone H3 binding. Even though treatment with an HDAC inhibitor produced the same effect as hsp90 inhibition, the effect of 17-AAG was independent of HDAC. We conclude that hsp90 inhibition attenuates NF-κB transcriptional activation by preventing coactivator recruitment and nucleosome eviction from the target promoter in human lung endothelial cells. PMID:24303801

A new mechanism of voltage-dependent gating exposed by KV10.1 channels interrupted between voltage sensor and pore.

Science.gov (United States)

Tomczak, Adam P; Fernández-Trillo, Jorge; Bharill, Shashank; Papp, Ferenc; Panyi, Gyorgy; Stühmer, Walter; Isacoff, Ehud Y; Pardo, Luis A

2017-05-01

Voltage-gated ion channels couple transmembrane potential changes to ion flow. Conformational changes in the voltage-sensing domain (VSD) of the channel are thought to be transmitted to the pore domain (PD) through an α-helical linker between them (S4-S5 linker). However, our recent work on channels disrupted in the S4-S5 linker has challenged this interpretation for the KCNH family. Furthermore, a recent single-particle cryo-electron microscopy structure of K V 10.1 revealed that the S4-S5 linker is a short loop in this KCNH family member, confirming the need for an alternative gating model. Here we use "split" channels made by expression of VSD and PD as separate fragments to investigate the mechanism of gating in K V 10.1. We find that disruption of the covalent connection within the S4 helix compromises the ability of channels to close at negative voltage, whereas disconnecting the S4-S5 linker from S5 slows down activation and deactivation kinetics. Surprisingly, voltage-clamp fluorometry and MTS accessibility assays show that the motion of the S4 voltage sensor is virtually unaffected when VSD and PD are not covalently bound. Finally, experiments using constitutively open PD mutants suggest that the presence of the VSD is structurally important for the conducting conformation of the pore. Collectively, our observations offer partial support to the gating model that assumes that an inward motion of the C-terminal S4 helix, rather than the S4-S5 linker, closes the channel gate, while also suggesting that control of the pore by the voltage sensor involves more than one mechanism. © 2017 Tomczak et al.

Enhancement of antibiotic activity by efflux inhibitors against multidrug resistant Mycobacterium tuberculosis clinical isolates from Brazil

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Tatiane eCoelho

2015-04-01

Full Text Available Drug resistant tuberculosis continues to increase and new approaches for its treatment are necessary. The identification of M. tuberculosis clinical isolates presenting efflux as part of their resistant phenotype has a major impact in tuberculosis treatment. In this work, we used a checkerboard procedure combined with the tetrazolium microplate-based assay (TEMA to study single combinations between antituberculosis drugs and efflux inhibitors (EIs against multidrug resistant M. tuberculosis clinical isolates using the fully susceptible strain H37Rv as reference. Efflux activity was studied on a real-time basis by a fluorometric method that uses ethidium bromide as efflux substrate. Quantification of efflux pump genes mRNA transcriptional levels were performed by RT-qPCR. The fractional inhibitory concentrations (FIC indicated synergistic activity for the interactions between isoniazid, rifampicin, amikacin, ofloxacin, and ethidium bromide plus the EIs verapamil, thioridazine and chlorpromazine. The FICs ranged from 0.25, indicating a four-fold reduction on the MICs, to 0.015, 64-fold reduction. The detection of active efflux by real-time fluorometry showed that all strains presented intrinsic efflux activity that contributes to the overall resistance which can be inhibited in the presence of the EIs. The quantification of the mRNA levels of the most important efflux pump genes on these strains shows that they are intrinsically predisposed to expel toxic compounds as the exposure to subinhibitory concentrations of antibiotics were not necessary to increase the pump mRNA levels when compared with the non-exposed counterpart. The results obtained in this study confirm that the intrinsic efflux activity contributes to the overall resistance in multidrug resistant clinical isolates of M. tuberculosis and that the inhibition of efflux pumps by the EIs can enhance the clinical effect of antibiotics that are their substrates.

Catecholamine levels in sheep hypothalamus, hypophysis and adrenals following whole-body gamma irradiation

International Nuclear Information System (INIS)

Pastorova, B.; Arendarcik, J.; Molnarova, M.

1985-01-01

Changes were studied in the levels of catecholamines and L-DOPA in the control system of the reproduction cycle (hypothalamus, hypophysis) and in the adrenal glands of sheep after whole-body irradiation with 60 Co at a total dose of 6.7 Gy for seven days. The output of the radiation source was 0.039 Gy/h. The catecholamines (noradrenaline, dopamine and adrenaline) and L-DOPA were determined after separation from the tissues by the method of spectral fluorometry. After whole-body exposure to gamma radiation, noradrenaline dropped in the hypothalamus in comparison with the control group, most significantly in the rostral (by 74.2%) and caudal (by 40%) parts. A similar drop was also observed in dopamine, the concentrations of which decreased in the rostral hypothalamus by 60%. Adrenaline showed a drop in the hypothalamus, most significant in the caudal region (by 62%). Consequently, the level of the precursor of the synthesis of catecholamines and L-DOPA changed and showed in the studied regions of the hypothalamus significantly lower levels than in the control group. As regards the hypophysis, after irradiation no significant changes in the levels of noradrenaline and adrenaline were recorded, however, dopamine and L-DOPA dropped significantly (P<0.01). The exposure to gamma radiation also causes a decrease in the concentrations of catecholamines and L-DOPA in the adrenal glands of sheep, most significantly in noradrenaline (by 61%). It was thus found that whole-body irradiation of sheep with a dose of 6.7 Gy results in a significant decrease in the level of catecholamines in the hypothalamus, hypophysis and adrenal glands, which is probably in relation to the failure of synthesis and degradation of catecholamines and to the total organism injury

Metabolic and hemodynamic activation of postischemic rat brain by cortical spreading depression.

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Kocher, M

1990-07-01

Following transient ischemia of the brain, the coupling between somatosensory activation and the hemodynamic-metabolic response is abolished for a certain period despite the partial recovery of somatosensory evoked responses. To determine whether this disturbance is due to alterations of the stimulus-induced neuronal excitation or to a breakdown of the coupling mechanisms, cortical spreading depression was used as a metabolic stimulus in rats before and after ischemia. Adult rats were subjected to 30 min of global forebrain ischemia and 3-6 h of recirculation. EEG, cortical direct current (DC) potential, and laser-Doppler flow were continuously recorded. Local CBF (LCBF), local CMRglc (LCMRglc), regional tissue contents of ATP, glucose, and lactate, and regional pH were determined by quantitative autoradiography, substrate-induced bioluminescence, and fluorometry. Amplitude and frequency of the DC shifts did not differ between groups. In control animals, spreading depression induced a 77% rise in cortical glucose consumption, a 66% rise in lactate content, and a drop in tissue pH of 0.3 unit. ATP and glucose contents were not depleted. During the passage of DC shifts, transient increases (less than 2 min) in laser-Doppler flow were observed, followed by a post-spreading depression hypoperfusion. A comparable although less expressed pattern of hemodynamic and metabolic changes was observed in the postischemic rats. Although baseline LCMRglc was depressed after ischemia, it was activated 47% during spreading depression. Lactate increased by 26%, pH decreased by 0.3 unit, and ATP and glucose remained unchanged. The extent of the transient increase in laser-Doppler flow did not differ from that of the control group, and a post-spreading depression hypoperfusion was also found.(ABSTRACT TRUNCATED AT 250 WORDS)

Differential effector responses by circulating/blood and tissue/peritoneal neutrophils following burn combined with Enterococcus faecalis infection.

Science.gov (United States)

Fazal, Nadeem; Shelip, Alla; Siddiqui, Erum; Ali, Ashraf; Azim, Anser C; Al-Ghoul, Walid M

2012-03-01

Recently we found that superimposition of Enterococcus faecalis infection on burn injury caused an eruption of host mortality not seen with either individual challenge. We hypothesized that the Enterococcus bacteria, and/or factors related to these organisms, aggravate burn-induced modulations in host defense by neutrophils. Our study focuses on alterations in neutrophils' oxidative, proteolytic, and adhesive functions and transendothelial migration of neutrophils in burn rats inoculated with E. faecalis. Rats were subjected to burn (30% total body surface area) and then intra-abdominally inoculated with E. faecalis (10(4)CFU kg(-1) b.w). Polymorphonuclear neutrophils (PMNs) were harvested from circulating/blood and tissue/peritoneal cavity at day-2 post injury. Extracellular release of O(-)(2) anion production was determined by luminometry, and intracellular production of reactive oxygen species was measured by digital imaging technique. Fluoroscan analysis and confocal microscopy determined intracellular elastase production. The expression of adhesion molecule CD11b/CD18 was performed by flow cytometry. Calcein AM-labeled PMNs were co-cultured with TNF-α-stimulated rat lung microvascular endothelial cells, and their ability to adhere was assessed by fluorometry and digital imaging and finally, chemotaxis was measured by neutrophil transmigration assays. The results showed differential effector responses by circulatory and/or tissue PMNs. Tissue/peritoneal PMNs produced more O(-)(2), less intracellular elastase, and increased expression of CD11b/CD18 accompanied with increased adhesivity of MIP-2-stimulated PMNs to endothelial cells as compared to circulatory/blood PMNs. This differential effect was more pronounced following burn plus E. faecalis infection, indicating that the combined injury changed neutrophil functions. © 2011 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.

Impact of a cyclonic eddy on phytoplankton community structure and photosynthetic competency in the subtropical North Pacific Ocean

Science.gov (United States)

Vaillancourt, Robert D.; Marra, John; Seki, Michael P.; Parsons, Michael L.; Bidigare, Robert R.

2003-07-01

A synoptic spatial examination of the eddy Haulani (17-20 November 2000) revealed a structure typical of Hawaiian cyclonic eddies with divergent surface flow forcing the upward displacement of deep waters. Hydrographic surveys revealed that surface water in the eddy center was ca. 3.5°C cooler, 0.5 saltier, and 1.4 kg m -3 denser than surface waters outside the eddy. Vertically integrated concentrations of nitrate+nitrite, phosphate and silicate were enhanced over out-eddy values by about 2-fold, and nitrate+nitrite concentrations were ca. 8× greater within the euphotic zone inside the eddy than outside. Si:N ratios were lower within the upper mixed layer of the eddy, indicating an enhanced Si uptake relative to nitrate+nitrite. Chlorophyll a concentrations were higher within the eddy compared to control stations outside, when integrated over the upper 150 m, but were not significantly different when integrated over the depth of the euphotic zone. Photosynthetic competency, assessed using fast repetition-rate fluorometry, varied with the doming of the isopycnals and the supply of macro-nutrients to the euphotic zone. The physical and chemical environment of the eddy selected for the accumulation of larger phytoplankton species. Photosynthetic bacteria ( Prochlorococcus and Synechococcus) and small (3 μm diameter) were more abundant inside the eddy than outside. Diatoms of the genera Rhizosolenia and Hemiaulus outside the eddy contained diazotrophic endosymbiontic cyanobacteria, but these endosymbionts were absent from the cells of these species inside the eddy. The increase in cell numbers of large photosynthetic eukaryotes with hard silica or calcite cell walls is likely to have a profound impact on the proportion of the organic carbon production that is exported to deep water by sinking of senescent cells and cells grazed by herbivorous zooplankton and repackaged as large fecal pellets.

Minocycline and doxycycline, but not other tetracycline-derived compounds, protect liver cells from chemical hypoxia and ischemia/reperfusion injury by inhibition of the mitochondrial calcium uniporter

International Nuclear Information System (INIS)

Schwartz, Justin; Holmuhamedov, Ekhson; Zhang, Xun; Lovelace, Gregory L.; Smith, Charles D.; Lemasters, John J.

2013-01-01

Minocycline, a tetracycline-derived compound, mitigates damage caused by ischemia/reperfusion (I/R) injury. Here, 19 tetracycline-derived compounds were screened in comparison to minocycline for their ability to protect hepatocytes against damage from chemical hypoxia and I/R injury. Cultured rat hepatocytes were incubated with 50 μM of each tetracycline-derived compound 20 min prior to exposure to 500 μM iodoacetic acid plus 1 mM KCN (chemical hypoxia). In other experiments, hepatocytes were incubated in anoxic Krebs–Ringer–HEPES buffer at pH 6.2 for 4 h prior to reoxygenation at pH 7.4 (simulated I/R). Tetracycline-derived compounds were added 20 min prior to reperfusion. Ca 2+ uptake was measured in isolated rat liver mitochondria incubated with Fluo-5N. Cell killing after 120 min of chemical hypoxia measured by propidium iodide (PI) fluorometry was 87%, which decreased to 28% and 42% with minocycline and doxycycline, respectively. After I/R, cell killing at 120 min decreased from 79% with vehicle to 43% and 49% with minocycline and doxycycline. No other tested compound decreased killing. Minocycline and doxycycline also inhibited mitochondrial Ca 2+ uptake and suppressed the Ca 2+ -induced mitochondrial permeability transition (MPT), the penultimate cause of cell death in reperfusion injury. Ru360, a specific inhibitor of the mitochondrial calcium uniporter (MCU), also decreased cell killing after hypoxia and I/R and blocked mitochondrial Ca 2+ uptake and the MPT. Other proposed mechanisms, including mitochondrial depolarization and matrix metalloprotease inhibition, could not account for cytoprotection. Taken together, these results indicate that minocycline and doxycycline are cytoprotective by way of inhibition of MCU. - Highlights: • Minocycline and doxycycline are the only cytoprotective tetracyclines of those tested • Cytoprotective tetracyclines inhibit the MPT and mitochondrial calcium and iron uptake. • Cytoprotective tetracyclines protect

Minocycline and doxycycline, but not other tetracycline-derived compounds, protect liver cells from chemical hypoxia and ischemia/reperfusion injury by inhibition of the mitochondrial calcium uniporter

Energy Technology Data Exchange (ETDEWEB)

Schwartz, Justin; Holmuhamedov, Ekhson; Zhang, Xun; Lovelace, Gregory L.; Smith, Charles D. [Department of Drug Discovery and Biomedical Sciences, Medical University of South Carolina, Charleston, SC (United States); Lemasters, John J., E-mail: JJLemasters@musc.edu [Department of Drug Discovery and Biomedical Sciences, Medical University of South Carolina, Charleston, SC (United States); Department of Biochemistry and Molecular Biology, Medical University of South Carolina, Charleston, SC (United States)

2013-11-15

Minocycline, a tetracycline-derived compound, mitigates damage caused by ischemia/reperfusion (I/R) injury. Here, 19 tetracycline-derived compounds were screened in comparison to minocycline for their ability to protect hepatocytes against damage from chemical hypoxia and I/R injury. Cultured rat hepatocytes were incubated with 50 μM of each tetracycline-derived compound 20 min prior to exposure to 500 μM iodoacetic acid plus 1 mM KCN (chemical hypoxia). In other experiments, hepatocytes were incubated in anoxic Krebs–Ringer–HEPES buffer at pH 6.2 for 4 h prior to reoxygenation at pH 7.4 (simulated I/R). Tetracycline-derived compounds were added 20 min prior to reperfusion. Ca{sup 2+} uptake was measured in isolated rat liver mitochondria incubated with Fluo-5N. Cell killing after 120 min of chemical hypoxia measured by propidium iodide (PI) fluorometry was 87%, which decreased to 28% and 42% with minocycline and doxycycline, respectively. After I/R, cell killing at 120 min decreased from 79% with vehicle to 43% and 49% with minocycline and doxycycline. No other tested compound decreased killing. Minocycline and doxycycline also inhibited mitochondrial Ca{sup 2+} uptake and suppressed the Ca{sup 2+}-induced mitochondrial permeability transition (MPT), the penultimate cause of cell death in reperfusion injury. Ru360, a specific inhibitor of the mitochondrial calcium uniporter (MCU), also decreased cell killing after hypoxia and I/R and blocked mitochondrial Ca{sup 2+} uptake and the MPT. Other proposed mechanisms, including mitochondrial depolarization and matrix metalloprotease inhibition, could not account for cytoprotection. Taken together, these results indicate that minocycline and doxycycline are cytoprotective by way of inhibition of MCU. - Highlights: • Minocycline and doxycycline are the only cytoprotective tetracyclines of those tested • Cytoprotective tetracyclines inhibit the MPT and mitochondrial calcium and iron uptake. • Cytoprotective

Sensitivity of the green algae Chlamydomonas reinhardtii to gamma radiation: Photosynthetic performance and ROS formation.

Science.gov (United States)

Gomes, Tânia; Xie, Li; Brede, Dag; Lind, Ole-Christian; Solhaug, Knut Asbjørn; Salbu, Brit; Tollefsen, Knut Erik

2017-02-01

The aquatic environment is continuously exposed to ionizing radiation from both natural and anthropogenic sources, making the characterization of ecological and health risks associated with radiation of large importance. Microalgae represent the main source of biomass production in the aquatic ecosystem, thus becoming a highly relevant biological model to assess the impacts of gamma radiation. However, little information is available on the effects of gamma radiation on microalgal species, making environmental radioprotection of this group of species challenging. In this context, the present study aimed to improve the understanding of the effects and toxic mechanisms of gamma radiation in the unicellular green algae Chlamydomonas reinhardtii focusing on the activity of the photosynthetic apparatus and ROS formation. Algal cells were exposed to gamma radiation (0.49-1677mGy/h) for 6h and chlorophyll fluorescence parameters obtained by PAM fluorometry, while two fluorescent probes carboxy-H 2 DFFDA and DHR 123 were used for the quantification of ROS. The alterations seen in functional parameters of C. reinhardtii PSII after 6h of exposure to gamma radiation showed modifications of PSII energy transfer associated with electron transport and energy dissipation pathways, especially at the higher dose rates used. Results also showed that gamma radiation induced ROS in a dose-dependent manner under both light and dark conditions. The observed decrease in photosynthetic efficiency seems to be connected to the formation of ROS and can potentially lead to oxidative stress and cellular damage in chloroplasts. To our knowledge, this is the first report on changes in several chlorophyll fluorescence parameters associated with photosynthetic performance and ROS formation in microalgae after exposure to gamma radiation. Copyright © 2016 Elsevier B.V. All rights reserved.

In vitro culture of pre-implanted mouse embryos. A model system for studying combined effects

International Nuclear Information System (INIS)

Streffer, C.; Beuningen, D. van; Molls, M.; Pon, A.; Schulz, S.; Zamboglou, N.

1978-01-01

Studies on combined effects, e.g. interaction between chemical toxicants and ionizing radiation, are difficult to perform, as they are dependent on many factors (substance concentration, radiation dose, sequence of treatments, etc.). In order to obtain data from such studies it is necessary to establish a comparatively simple experimental model system. We have established such a model system by studying combined effects on pre-implanted mouse embryos cultured in vitro. This system has the following advantages: (1) The embryos can be cultivated for several days in vitro; (2) Their physiological intactness can be tested; and (3) Cell proliferation, cell killing and chromosomal damage can be investigated comparatively easily. The embryos are isolated at the 2-cell stage and incubated in a culture medium in vitro. The development of the embryos is followed under the microscope until the development of blastocysts or the hatching of blastocysts is observed. These blastocysts can be transplanted to fostered mice and the development of normal animals determined. The proliferation kinetics can be studied easily, and the methods are described. A method has also been developed to measure the DNA content of individual cells by microscope fluorometry. After treatment of the embryos with ionizing radiation or drugs the release of micronuclei has been observed from the cell nuclei, which is an expression for chromosomal damage. Substances or radionuclides can be added to the culture medium or external irradiation can be performed during the culture period. Also the combined effects of radiation and heating can be studied. The effects of X-rays and tritiated compounds have also been investigated. The combined effects of radiation with antibiotics such as actinomycin D, and environmental toxicants such as lead, have been determined. The system described has been useful to evaluate cytological, teratogenic and cytogenetic effects

Feeding sustains photosynthetic quantum yield of a scleractinian coral during thermal stress.

Science.gov (United States)

Borell, Esther M; Bischof, Kai

2008-10-01

Thermal resistance of the coral-zooxanthellae symbiosis has been associated with chronic photoinhibition, increased antioxidant activity and protein repair involving high demands of nitrogen and energy. While the relative importance of heterotrophy as a source of nutrients and energy for cnidarian hosts, and as a means of nitrogen acquisition for their zooxanthellae, is well documented, the effect of feeding on the thermal sensitivity of the symbiotic association has been so far overlooked. Here we examine the effect of zooplankton feeding versus starvation on the bleaching susceptibility and photosynthetic activity of photosystem II (PSII) of zooxanthellae in the scleractinian coral Stylophora pistillata in response to thermal stress (daily temperature rises of 2-3 degrees C) over 10 days, employing pulse-amplitude-modulated chlorophyll fluorometry. Fed and starved corals displayed a decrease in daily maximum potential quantum yield (F (v)/F (m)) of PSII, effective quantum yield (F/F (m)') and relative electron transport rates over the course of 10 days. However after 10 days of exposure to elevated temperature, F (v)/F (m) of fed corals was still 50-70% higher than F (v)/F (m) of starved corals. Starved corals showed strong signs of chronic photoinhibition, which was reflected in a significant decline in nocturnal recovery rates of PSII relative to fed corals. This was paralleled by the progressive inability to dissipate excess excitation energy via non-photochemical quenching (NPQ). After 10 days, NPQ of starved corals had decreased by about 80% relative to fed corals. Feeding treatment had no significant effect on chlorophyll a and c (2) concentrations and zooxanthellae densities, but the mitotic indices were significantly lower in starved than in fed corals. Collectively the results indicate that exogenous food may reduce the photophysiological damage of zooxanthellae that typically leads to bleaching and could therefore play an important role in mediating the

Non-intrusive Assessment of Photosystem II and Photosystem I in Whole Coral Tissues

Directory of Open Access Journals (Sweden)

Milán Szabó

2017-08-01

Full Text Available Reef building corals (phylum Cnidaria harbor endosymbiotic dinoflagellate algae (genus Symbiodinium that generate photosynthetic products to fuel their host's metabolism. Non-invasive techniques such as chlorophyll (Chl fluorescence analyses of Photosystem II (PSII have been widely used to estimate the photosynthetic performance of Symbiodinium in hospite. However, since the spatial origin of PSII chlorophyll fluorescence in coral tissues is uncertain, such signals give limited information on depth-integrated photosynthetic performance of the whole tissue. In contrast, detection of absorbance changes in the near infrared (NIR region integrates signals from deeper tissue layers due to weak absorption and multiple scattering of NIR light. While extensively utilized in higher plants, NIR bio-optical techniques are seldom applied to corals. We have developed a non-intrusive measurement method to examine photochemistry of intact corals, based on redox kinetics of the primary electron donor in Photosystem I (P700 and chlorophyll fluorescence kinetics (Fast-Repetition Rate fluorometry, FRRf. Since the redox state of P700 depends on the operation of both PSI and PSII, important information can be obtained on the PSII-PSI intersystem electron transfer kinetics. Under moderate, sub-lethal heat stress treatments (33°C for ~20 min, the coral Pavona decussata exhibited down-regulation of PSII electron transfer kinetics, indicated by slower rates of electron transport from QA to plastoquinone (PQ pool, and smaller relative size of oxidized PQ with concomitant decrease of a specifically-defined P700 kinetics area, which represents the active pool of PSII. The maximum quantum efficiency of PSII (Fv/Fm and functional absorption cross-section of PSII (σPSII remained unchanged. Based on the coordinated response of P700 parameters and PSII-PSI electron transport properties, we propose that simple P700 kinetics parameters as employed here serve as indicators of

The microbial mats of Pavilion Lake microbialites: examining the relationship between photosynthesis and carbonate precipitation

Science.gov (United States)

Lim, D. S. S.; Hawes, I.; Mackey, T. J.; Brady, A. L.; Biddle, J.; Andersen, D. T.; Belan, M.; Slater, G.; Abercromby, A.; Squyres, S. W.; Delaney, M.; Haberle, C. W.; Cardman, Z.

2014-12-01

Pavilion Lake in British Columbia, Canada is an ultra-oligotrophic lake that has abundant microbialite growth. Recent research has shown that photoautotrophic microbial communities are important to modern microbialite development in Pavilion Lake. However, questions remain as to the relationship between changing light levels within the lake, variation in microbialite macro-structure, microbial consortia, and the preservation of associated biosignatures within the microbialite fabrics. The 2014 Pavilion Lake Research Project (PLRP) field program was focused on data gathering to understand these complex relationships by determining if a) light is the immediate limit to photosynthetic activity and, if so, if light is distributed around microbialites in ways that are consistent with emergent microbialite structure; and b) if at more local scales, the filamentous pink and green cyanobacterial nodular colonies identified in previous PLRP studies are centers of photosynthetic activity that create pH conditions suitable for carbonate precipitation. A diver-deployed pulse-amplitude modulated (PAM) fluorometer was used to collect synoptic in situ measurements of fluorescence yield and irradiance and across microbialites, focusing on comparing flat and vertical structural elements at a range of sites and depths. As well, we collected time series measurements of photosynthetic activity and irradiance at a set depth of 18 m across three different regions in Pavilion Lake. Our initial findings suggest that all microbialite surfaces are primarily light-limited regardless of depth or location within the lake. Shore based PAM fluorometry and microelectrode profiling of diver-collected samples suggest that pink and green nodules have different photosynthetic properties and pH profiles, and that nodular growth is likely to be the primary route of calcification due to the gelatinous covering the nodule creates. On-going tests for molecular signatures and isotopic shifts will allow for

Uranium hydrogeochemical and stream sediment reconnaissance data release for the Rock Springs NTMS Quadrangle, Wyoming, including concentrations of forty-two additional elements

International Nuclear Information System (INIS)

Morgan, T.L.

1981-01-01

This report contains data collected by the Los Alamos Scientific Laboratory (LASL) during a regional geochemical survey for uranium in the Rock Springs National Topographic Map Series (NTMS) quadrangle, southwestern Wyoming, as part of the nationwide hydrogeochemical and Stream Sediment Reconnaissance (HSSR). Totals of 397 water and 1794 sediment samples were collected from 1830 locations in the Rock Springs quadrangle of southern Wyoming during the summer of 1976. The average uranium concentration of all water samples is 6.57 ppb and the average sediment uranium concentration is 3.64 ppM. Elemental concentration, field measurement, weather, geologic, and geographic data for each sample location are listed for waters and for sediments in the appendices. Uranium/thorium ratios for sediment samples are also included. A sample location overlay (Plate I) at 1:250 000 scale for use in conjunction with the Rock Springs NTMS quadrangle sheet (US Geological Survey, 1954) is provided. All elemental analyses were performed at the LASL. Water samples were initially analyzed for uranium by fluorometry. All water samples containing more than 40 ppB uranium were reanalyzed by delayed-neutron counting. Sediments were analyzed for uranium and thorium as well as Al, Sb, Ba, Be, Bi, Cd, Ca, Ce, Cs, Cl, Cr, Co, Cu, Dy, Eu, Au, Hf, Fe, La, Pb, Li, Lu, Mg, Mn, Ni, Nb, K, Rb, Sm, Sc, Ag, Na, Sr, Ta, Tb, Sn, T, W, V, Yb, and Zn. All sediments were analyzed for uranium by delayed-neutron counting. Other elemental concentrations in sediments were determined by neutron-activation analysis for 30 elements, by x-ray fluorescence for 12 elements, and by arc-source emission spectrography for 2 elements. These analytical methods are described briefly in the appendix. This report is simply a data release and is intended to make the data available to the DOE and to the public as quickly as possible

Changes in T cell populations due to local irradiation of a portion of the maxilla in mice

International Nuclear Information System (INIS)

Satoh, Daigo

1997-01-01

The aim of this study was to investigate the changes in the immune organs after head and neck irradiation. The numbers of lymphocytes in peripheral blood, the spleen and the thymus following local irradiation of a portion of the maxilla in mice were studied using three-color fluorometry and were compared with a non-irradiation group. In the peripheral blood, the absolute numbers of T cells, CD4 + SP cells and CD8 + SP cells decreased after irradiation, and the period of the decrease was longer than the decreases in number of leukocytes and lymphocytes. The ratio of CD4 + SP cells showed a significant decrease, and the ratio of CD8+ S P cells showed a significant increase 1 day after irradiation. In the spleen, the absolute number of T cells, the radio of CD4 + SP and CD8 + SP cell subsets showed a decrease, and the period of the decrease was longer than the decrease of the wet-weight of the spleen, and also longer than the decrease of the number of leukocytes. The number of CD4 + SP cells showed a signigicant increase, and CD8 + SP cells showed a significant decrease 21 days after irradiation. In the thymus, the absolute number of TCR αβ-thymocytes did not show a significant decrease. However, the number of DN thymocytes showed a marked decrease. These results indicate that the numbers of T cells in peripheral blood, the spleen and the thymus change immediately after irradiation, and the numbers of lymphocytes and the T cells in the spleen recover more slowly than that in the peripheral blood. As lymphoid tissues showed the suppression of immunological response for a long period, it was suggested that lymphoid tissues have to be observed carefully after irradiation to prevent cancer metastasis. (K.H.)

Biotin increases glucokinase expression via soluble guanylate cyclase/protein kinase G, adenosine triphosphate production and autocrine action of insulin in pancreatic rat islets.

Science.gov (United States)

Vilches-Flores, Alonso; Tovar, Armando R; Marin-Hernandez, Alvaro; Rojas-Ochoa, Alberto; Fernandez-Mejia, Cristina

2010-07-01

Besides its role as a carboxylase prosthetic group, biotin has important effects on gene expression. However, the molecular mechanisms through which biotin exerts these effects are largely unknown. We previously found that biotin increases pancreatic glucokinase expression. We have now explored the mechanisms underlying this effect. Pancreatic islets from Wistar rats were treated with biotin, in the presence or absence of different types of inhibitors. Glucokinase mRNA and 18s rRNA abundance were determined by real-time PCR. Adenosine triphosphate (ATP) content was analyzed by fluorometry. Biotin treatment increased glucokinase mRNA abundance approximately one fold after 2 h; the effect was sustained up to 24 h. Inhibition of soluble guanylate cyclase or protein kinase G (PKG) signalling suppressed biotin-induced glucokinase expression. The cascade of events downstream of PKG in biotin-mediated gene transcription is not known. We found that inhibition of insulin secretion with diazoxide or nifedipine prevented biotin-stimulated glucokinase mRNA increase. Biotin treatment increased islet ATP content (control: 4.68+/-0.28; biotin treated: 6.62+/-0.26 pmol/islet) at 30 min. Inhibition of PKG activity suppressed the effects of biotin on ATP content. Insulin antibodies or inhibitors of phosphoinositol-3-kinase/Akt insulin signalling pathway prevented biotin-induced glucokinase expression. The nucleotide 8-Br-cGMP mimicked the biotin effects. We propose that the induction of pancreatic glucokinase mRNA by biotin involves guanylate cyclase and PKG activation, which leads to an increase in ATP content. This induces insulin secretion via ATP-sensitive potassium channels. Autocrine insulin, in turn, activates phosphoinositol-3-kinase/Akt signalling. Our results offer new insights into the pathways that participate in biotin-mediated gene expression. (c) 2010 Elsevier Inc. All rights reserved.

Small Moves, NUI. Small Moves: Beginning to Investigate Biogeochemical Exchange From the Seafloor to the Exterior of an Ice-Covered Ocean

Science.gov (United States)

German, C. R.; Boetius, A.

2017-12-01

We present results from two recent cruises, using the new Nereid Under Ice (NUI) vehicle aboard the FS Polarstern, in which we investigated biogeochemical fluxes from the deep seafloor of the Gakkel Ridge, an ultraslow spreading ridge that spans the ice-covered Arctic Ocean, and the mechanisms by which biogeochemical signals might be transferred from within the underlying ocean to the overlying Arctic ice. The scientific advances for this work progress hand in hand with technological capability. During a first cruise in 2014, our NUI-based investigations focused on photosynthetically-driven biogeochemical cycling in the uppermost water column and how to study such processes using in situ sensing immediately at and beneath the rough topography of the overlying ice-cover. For that work we relied entirely upon human-in-the-loop control of the vehicle via a single optical fiber light tether than provided real-time monitoring and control of the vehicle as it ranged laterally out under the ice up to 1km distant from the ship, conducting physical, geochemical and biological surveys. Instrumentation used for that work included multibeam mapping and imaging (digital still photographs and HD video), in situ spectroscopy to study light transmission through the ice and biogeochemical mapping of the ocean water column using a combination of CTD sensing, fluorometry and an in situ nitrate analyzer. Returning to the Arctic in 2016 we extended our exploration modes with NUI further, investigating for seafloor fluid flow at a shallow setting on the flanks of the Gakkel Ridge where the seabed rises from >4000m to movement of the ship (horizontal displacements of 1km or more) at the ice-covered ocean surface. While the existing NUI vehicle does not map directly to model payloads for future SLS missions to Europa or Enceladus it does provide for important small moves in the right direction.

Toxic pressure of herbicides on microalgae in Dutch estuarine and coastal waters

Science.gov (United States)

Booij, Petra; Sjollema, Sascha B.; van der Geest, Harm G.; Leonards, Pim E. G.; Lamoree, Marja H.; de Voogt, W. Pim; Admiraal, Wim; Laane, Remi W. P. M.; Vethaak, A. Dick

2015-08-01

For several decades now, there has been an increase in the sources and types of chemicals in estuarine and coastal waters as a consequence of anthropogenic activities. This has led to considerable concern about the effects of these chemicals on the marine food chain. The fact is that estuarine and coastal waters are the most productive ecosystems with high primary production by microalgae. The toxic pressure of specific phytotoxic chemicals now poses a major threat to these ecosystems. In a previous study, six herbicides (atrazine, diuron, irgarol, isoproturon, terbutryn and terbutylazine) were identified as the main contaminants affecting photosynthesis in marine microalgae. The purpose of this study is to investigate the toxic pressure of these herbicides in the Dutch estuarine and coastal waters in relation to the effective photosystem II efficiency (ΦPSII) in microalgae. Temporal and spatial variations in the concentrations of these herbicides were analyzed based on monitoring data. Additionally, a field study was carried out in which chemical analysis of water was performed and also a toxicity assessment using the Pulse Amplitude Modulation (PAM) fluorometry assay that measures ΦPSII. The toxic pressure on ΦPSII in microalgae has decreased with 55-82% from 2003 to 2012, with the Western Scheldt estuary showing the highest toxic pressure. By combining toxicity data from the PAM assay with chemical analysis of herbicide concentrations, we have identified diuron and terbutylazine as the main contributors to the toxic pressure on microalgae. Although direct effects are not expected, the toxic pressure is close to the 10% effect level in the PAM assay. A compliance check with the current environmental legislation of the European Union revealed that the quality standards are not sufficient to protect marine microalgae.

Estimating phytoplankton photosynthesis by active fluorescence

Energy Technology Data Exchange (ETDEWEB)

Falkowski, P.G.; Kolber, Z.

1992-01-01

Photosynthesis can be described by target theory, At low photon flux densities, photosynthesis is a linear function of irradiance (I), The number of reaction centers (n), their effective absorption capture cross section {sigma}, and a quantum yield {phi}. As photosynthesis becomes increasingly light saturated, an increased fraction of reaction centers close. At light saturation the maximum photosynthetic rate is given as the product of the number of reaction centers (n) and their maximum electron transport rate (I/{tau}). Using active fluorometry it is possible to measure non-destructively and in real time the fraction of open or closed reaction centers under ambient irradiance conditions in situ, as well as {sigma} and {phi} {tau} can be readily, calculated from knowledge of the light saturation parameter, I{sub k} (which can be deduced by in situ by active fluorescence measurements) and {sigma}. We built a pump and probe fluorometer, which is interfaced with a CTD. The instrument measures the fluorescence yield of a weak probe flash preceding (f{sub 0}) and succeeding (f{sub 0}) a saturating pump flash. Profiles of the these fluorescence yields are used to derive the instantaneous rate of gross photosynthesis in natural phytoplankton communities without any incubation. Correlations with short-term simulated in situ radiocarbon measurements are extremely high. The average slope between photosynthesis derived from fluorescence and that measured by radiocarbon is 1.15 and corresponds to the average photosynthetic quotient. The intercept is about 15% of the maximum radiocarbon uptake and corresponds to the average net community respiration. Profiles of photosynthesis and sections showing the variability in its composite parameters reveal a significant effect of nutrient availability on biomass specific rates of photosynthesis in the ocean.

Estimating phytoplankton photosynthesis by active fluorescence

Energy Technology Data Exchange (ETDEWEB)

Falkowski, P.G.; Kolber, Z.

1992-10-01

Photosynthesis can be described by target theory, At low photon flux densities, photosynthesis is a linear function of irradiance (I), The number of reaction centers (n), their effective absorption capture cross section {sigma}, and a quantum yield {phi}. As photosynthesis becomes increasingly light saturated, an increased fraction of reaction centers close. At light saturation the maximum photosynthetic rate is given as the product of the number of reaction centers (n) and their maximum electron transport rate (I/{tau}). Using active fluorometry it is possible to measure non-destructively and in real time the fraction of open or closed reaction centers under ambient irradiance conditions in situ, as well as {sigma} and {phi} {tau} can be readily, calculated from knowledge of the light saturation parameter, I{sub k} (which can be deduced by in situ by active fluorescence measurements) and {sigma}. We built a pump and probe fluorometer, which is interfaced with a CTD. The instrument measures the fluorescence yield of a weak probe flash preceding (f{sub 0}) and succeeding (f{sub 0}) a saturating pump flash. Profiles of the these fluorescence yields are used to derive the instantaneous rate of gross photosynthesis in natural phytoplankton communities without any incubation. Correlations with short-term simulated in situ radiocarbon measurements are extremely high. The average slope between photosynthesis derived from fluorescence and that measured by radiocarbon is 1.15 and corresponds to the average photosynthetic quotient. The intercept is about 15% of the maximum radiocarbon uptake and corresponds to the average net community respiration. Profiles of photosynthesis and sections showing the variability in its composite parameters reveal a significant effect of nutrient availability on biomass specific rates of photosynthesis in the ocean.

Covalent attachment of the plant natural product naringenin to small glass and ceramic beads

Directory of Open Access Journals (Sweden)

Grotewold Erich

2005-10-01

Full Text Available Abstract Background Natural products have numerous medicinal applications and play important roles in the biology of the organisms that accumulate them. Few methods are currently available for identifying proteins that bind to small molecules, therefore the discovery of cellular targets for natural products with pharmacological activity continues to pose a significant challenge in drug validation. Similarly, the identification of enzymes that participate in the biosynthesis or modification of natural products remains a formidable bottleneck for metabolic engineering. Flavonoids are one large group of natural products with a diverse number of functions in plants and in human health. The coupling of flavonoids to small ceramic and glass beads provides a first step in the development of high-throughput, solid-support base approaches to screen complex libraries to identify proteins that bind natural products. Results The utilization of small glass and ceramic beads as solid supports for the coupling of small molecules was explored. Initial characterization of the beads indicated uniform and high capacity loading of amino groups. Once the beads were deemed adequate for the linking of small molecules by the coupling of NHS-fluorescein followed by microscopy, chemical hydrolysis and fluorometry, the flavonoid naringenin was modified with 1,4-dibromobutane, followed by the attachment of aminopropyltriethoxysilane. After NMR structural confirmation, the resulting 7-(4-(3-(triethoxysilylpropylaminobutoxy naringenin was attached to the ceramic beads. Conclusion Our results demonstrate that ceramic and glass beads provide convenient solid supports for the efficient and facile coupling of small molecules. We succeeded in generating naringenin-coupled ceramic and glass beads. We also developed a convenient series of steps that can be applied for the solid-support coupling of other related flavonoids. The availability of solid-support coupled naringenin opens

Tensiometric, fluorescence and 1H NMR study of mixed micellization of non-steroidal anti-inflammatory drug sodium salt of ibuprofen in the presence of non-ionic surfactant in aqueous/urea solutions

International Nuclear Information System (INIS)

Rub, Malik Abdul; Khan, Farah; Sheikh, Mohmad Shafi; Azum, Naved; Asiri, Abdullah M.

2016-01-01

Highlights: • Interaction between (IBF + TX-100) mixtures has been investigated. • (IBF + TX-100) mixtures exhibit synergistic behavior. • Urea increases the surface charge of the micelles resulting halt of the micelles formation. • N agg , K sv and dielectric constant of mixed systems have also been evaluated. • 1 H NMR data suggested that IBF and TX-100 interacts through hydrophobic as well as hydrophillic interaction. - Abstract: The desirable surface/bulk properties for specific applications of drug sodium salt of ibuprofen (IBF) and Triton X-100 (TX-100) can be achieved by adjusting mainly the composition of these systems. The interactions of anionic drug IBF with non-ionic surfactant TX-100 micelles have been investigated using tensiometry, fluorometry and 1 H NMR in aqueous as well in 250 mmol⋅kg −1 urea solutions. Different theoretical models like Clint, Rubingh, and Rosen, etc. were utilized to get information about the nature of interaction between these two in bulk and at the interface. These models disclose that the non-ideal behavior with attractive interaction in bulk and at the interface exists. The steady-state fluorescence quenching study was employed to evaluate micelle aggregation numbers (N agg ), which signify the involvement of surfactant was forever higher compared to IBF. Stern–Volmer binding constants (K sv ), micropolarity (I 1 /I 3 ) and dielectric constant (D exp ) of the mixtures are also obtained using fluorescence method. By the addition of urea raise in the surface charge of the micelles was observed followed by halt of the micellization of drug and surfactant as well as their mixture, therefore cmc values increases followed by decrease in aggregation number. The 1 H NMR resonance intensity variations were paralleled by upfield shifts in the resonance frequencies, due to an increased shielding of IBF happening from closeness of the non-ionic TX-100 surfactant.

Measuring Photosynthetic Response to Drought Stress using Active and Passive Fluorescence

Science.gov (United States)

Helm, L.; Lerdau, M.; Wang, W.; Yang, X.

2017-12-01

Photosynthesis, the endothermic reactions involving the absorption of light and fixation and reduction of carbon dioxide by plants, plays important roles in carbon and water cycles, food security, and even weather and climate patterns. Solar radiation provides the energy for photosynthesis, but often plants absorb more solar energy than they can use to reduce carbon dioxide. This excess energy, which is briefly stored as high-energy electrons in the chloroplast, must be removed or damage to the leaf's photosynthetic machinery will occur. One important energy dissipation pathway is for the high energy electrons to return to their lower valance state and, in doing so, release radiation (fluorescence). This fluorescence (known as solar induced fluorescence (SIF) has been found to strongly correlate with gross photosynthesis. Recent advances in the remote sensing of SIF allow for large-scale real-time estimation of photosynthesis. In a warming climate with more frequent stress, remote sensing is necessary for measuring the spatial and temporal variability of photosynthesis. However, the mechanisms that link SIF and photosynthesis are unclear, particularly how the relationship may or may not change under stress. We present data from leaf-level measurements of gas exchange, pulse amplitude modulation (PAM) fluorescence, and SIF in two major tree species in North America. Water-stressed and well-watered plants were compared to determine how SIF and carbon dioxide exchange are modulated by drought diurnally and seasonally. Secondly, photosynthesis and fluorescence under high and low oxygen concentrations were compared to determine how photorespiration alters the relationship between SIF and gross photosynthesis. We find a strong correlation between SIF and steady-state fluorescence measured with conventional PAM fluorometry. Our results also indicate that drought-stress modulates the SIF-photosynthesis relationship, and this may be driven by drought-induced changes in

Wilted cucumber plants infected by Fusarium oxysporum f. sp. cucumerinum do not suffer from water shortage.

Science.gov (United States)

Sun, Yuming; Wang, Min; Li, Yingrui; Gu, Zechen; Ling, Ning; Shen, Qirong; Guo, Shiwei

2017-09-01

Fusarium wilt is primarily a soil-borne disease and results in yield loss and quality decline in cucumber (Cucumis sativus). The main symptom of fusarium wilt is the wilting of entire plant, which could be caused by a fungal toxin(s) or blockage of water transport. To investigate whether this wilt arises from water shortage, the physiological responses of hydroponically grown cucumber plants subjected to water stress using polyethylene glycol (PEG, 6000) were compared with those of plants infected with Fusarium oxysporum f. sp. cucumerinum (FOC). Parameters reflecting plant water status were measured 8d after the start of treatment. Leaf gas exchange parameters and temperature were measured with a LI-COR portable open photosynthesis system and by thermal imaging. Chlorophyll fluorescence and chloroplast structures were assessed by imaging pulse amplitude-modulated fluorometry and transmission electron microscopy, respectively. Cucumber water balance was altered after FOC infection, with decreased water absorption and hydraulic conductivity. However, the responses of cucumber leaves to FOC and PEG differed in leaf regions. Under water stress, measures of lipid peroxidation (malondialdehyde) and chlorophyll fluorescence indicated that the leaf edge was more seriously injured, with a higher leaf temperature and disrupted leaf water status compared with the centre. Here, abscisic acid (ABA) and proline were negatively correlated with water potential. In contrast, under FOC infection, membrane damage and a higher temperature were observed in the leaf centre while ABA and proline did not vary with water potential. Cytologically, FOC-infected cucumber leaves exhibited circular chloroplasts and swelled starch grains in the leaf centre, in which they again differed from PEG-stressed cucumber leaves. This study illustrates the non-causal relationship between fusarium wilt and water transport blockage. Although leaf wilt occurred in both water stress and FOC infection, the

Natural lipid extracts and biomembrane-mimicking lipid compositions are disposed to form nonlamellar phases, and they release DNA from lipoplexes most efficiently

Energy Technology Data Exchange (ETDEWEB)

Koynova, Rumiana; MacDonald, Robert C. (NWU)

2010-01-18

A viewpoint now emerging is that a critical factor in lipid-mediated transfection (lipofection) is the structural evolution of lipoplexes upon interacting and mixing with cellular lipids. Here we report our finding that lipid mixtures mimicking biomembrane lipid compositions are superior to pure anionic liposomes in their ability to release DNA from lipoplexes (cationic lipid/DNA complexes), even though they have a much lower negative charge density (and thus lower capacity to neutralize the positive charge of the lipoplex lipids). Flow fluorometry revealed that the portion of DNA released after a 30-min incubation of the cationic O-ethylphosphatidylcholine lipoplexes with the anionic phosphatidylserine or phosphatidylglycerol was 19% and 37%, respectively, whereas a mixture mimicking biomembranes (MM: phosphatidylcholine/phosphatidylethanolamine/phosphatidylserine /cholesterol 45:20:20:15 w/w) and polar lipid extract from bovine liver released 62% and 74%, respectively, of the DNA content. A possible reason for this superior power in releasing DNA by the natural lipid mixtures was suggested by structural experiments: while pure anionic lipids typically form lamellae, the natural lipid mixtures exhibited a surprising predilection to form nonlamellar phases. Thus, the MM mixture arranged into lamellar arrays at physiological temperature, but began to convert to the hexagonal phase at a slightly higher temperature, {approx} 40-45 C. A propensity to form nonlamellar phases (hexagonal, cubic, micellar) at close to physiological temperatures was also found with the lipid extracts from natural tissues (from bovine liver, brain, and heart). This result reveals that electrostatic interactions are only one of the factors involved in lipid-mediated DNA delivery. The tendency of lipid bilayers to form nonlamellar phases has been described in terms of bilayer 'frustration' which imposes a nonzero intrinsic curvature of the two opposing monolayers. Because the stored

Short-term regulation and alternative pathways of photosynthetic electron transport in Hibiscus rosa-sinensis leaves.

Science.gov (United States)

Trubitsin, Boris V; Vershubskii, Alexey V; Priklonskii, Vladimir I; Tikhonov, Alexander N

2015-11-01

In this work, using the EPR and PAM-fluorometry methods, we have studied induction events of photosynthetic electron transport in Hibiscus rosa-sinensis leaves. The methods used are complementary, providing efficient tools for in situ monitoring of P700 redox transients and photochemical activity of photosystem II (PSII). The induction of P700(+) in dark-adapted leaves is characterized by the multiphase kinetics with a lag-phase, which duration elongates with the dark-adaptation time. Analyzing effects of the uncoupler monensin and artificial electron carrier methylviologen (MV) on photooxidation of P700 and slow induction of chlorophyll a fluorescence (SIF), we could ascribe different phases of transient kinetics of electron transport processes in dark-adapted leaves to the following regulatory mechanisms: (i) acceleration of electron transfer on the acceptor side of PSI, (ii) pH-dependent modulation of the intersystem electron flow, and (iii) re-distribution of electron fluxes between alternative (linear, cyclic, and pseudocyclic) pathways. Monensin significantly decreases a level of P700(+) and inhibits SIF. MV, which mediates electron flow from PSI to O2 with consequent formation of H2O2, promotes a rapid photooxidation of P700 without any lag-phase peculiar to untreated leaves. MV-mediated water-water cycle (H2O→PSII→PSI→MV→O2→H2O2→H2O) is accompanied by generation of ascorbate free radicals. This suggests that the ascorbate peroxidase system of defense against reactive oxygen species is active in chloroplasts of H. rosa-sinensis leaves. In DCMU-treated chloroplasts with inhibited PSII, the contribution of cyclic electron flow is insignificant as compared to linear electron flow. For analysis of induction events, we have simulated electron transport processes within the framework of our generalized mathematical model of oxygenic photosynthesis, which takes into account pH-dependent mechanisms of electron transport control and re-distribution of

Interplay between Mutations and Efflux in Drug Resistant Clinical Isolates of Mycobacterium tuberculosis

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Miguel Viveiros

2017-04-01

Full Text Available Numerous studies show efflux as a universal bacterial mechanism contributing to antibiotic resistance and also that the activity of the antibiotics subject to efflux can be enhanced by the combined use of efflux inhibitors. Nevertheless, the contribution of efflux to the overall drug resistance levels of clinical isolates of Mycobacterium tuberculosis is poorly understood and still is ignored by many. Here, we evaluated the contribution of drug efflux plus target-gene mutations to the drug resistance levels in clinical isolates of M. tuberculosis. A panel of 17 M. tuberculosis clinical strains were characterized for drug resistance associated mutations and antibiotic profiles in the presence and absence of efflux inhibitors. The correlation between the effect of the efflux inhibitors and the resistance levels was assessed by quantitative drug susceptibility testing. The bacterial growth/survival vs. growth inhibition was analyzed through the comparison between the time of growth in the presence and absence of an inhibitor. For the same mutation conferring antibiotic resistance, different MICs were observed and the different resistance levels found could be reduced by efflux inhibitors. Although susceptibility was not restored, the results demonstrate the existence of a broad-spectrum synergistic interaction between antibiotics and efflux inhibitors. The existence of efflux activity was confirmed by real-time fluorometry. Moreover, the efflux pump genes mmr, mmpL7, Rv1258c, p55, and efpA were shown to be overexpressed in the presence of antibiotics, demonstrating the contribution of these efflux pumps to the overall resistance phenotype of the M. tuberculosis clinical isolates studied, independently of the genotype of the strains. These results showed that the drug resistance levels of multi- and extensively-drug resistant M. tuberculosis clinical strains are a combination between drug efflux and the presence of target-gene mutations, a reality

Voltage-sensing phosphatase modulation by a C2 domain.

Science.gov (United States)

Castle, Paul M; Zolman, Kevin D; Kohout, Susy C

2015-01-01

The voltage-sensing phosphatase (VSP) is the first example of an enzyme controlled by changes in membrane potential. VSP has four distinct regions: the transmembrane voltage-sensing domain (VSD), the inter-domain linker, the cytosolic catalytic domain, and the C2 domain. The VSD transmits the changes in membrane potential through the inter-domain linker activating the catalytic domain which then dephosphorylates phosphatidylinositol phosphate (PIP) lipids. The role of the C2, however, has not been established. In this study, we explore two possible roles for the C2: catalysis and membrane-binding. The Ci-VSP crystal structures show that the C2 residue Y522 lines the active site suggesting a contribution to catalysis. When we mutated Y522 to phenylalanine, we found a shift in the voltage dependence of activity. This suggests hydrogen bonding as a mechanism of action. Going one step further, when we deleted the entire C2 domain, we found voltage-dependent enzyme activity was no longer detectable. This result clearly indicates the entire C2 is necessary for catalysis as well as for modulating activity. As C2s are known membrane-binding domains, we tested whether the VSP C2 interacts with the membrane. We probed a cluster of four positively charged residues lining the top of the C2 and suggested by previous studies to interact with phosphatidylinositol 4,5-bisphosphate [PI(4,5)P2] (Kalli et al., 2014). Neutralizing those positive charges significantly shifted the voltage dependence of activity to higher voltages. We tested membrane binding by depleting PI(4,5)P2 from the membrane using the 5HT2C receptor and found that the VSD motions as measured by voltage clamp fluorometry (VCF) were not changed. These results suggest that if the C2 domain interacts with the membrane to influence VSP function it may not occur exclusively through PI(4,5)P2. Together, this data advances our understanding of the VSP C2 by demonstrating a necessary and critical role for the C2 domain in

Sensitivity of the green algae Chlamydomonas reinhardtii to gamma radiation: Photosynthetic performance and ROS formation

Energy Technology Data Exchange (ETDEWEB)

Gomes, Tânia, E-mail: tania.gomes@niva.no [Norwegian Institute for Water Research (NIVA), Section of Ecotoxicology and Risk Assessment, Gaustadalléen 21, N-0349, Oslo (Norway); Centre for Environmental Radioactivity, Norwegian University of Life Sciences (NMBU), Post Box 5003, N-1432 Ås (Norway); Xie, Li [Norwegian Institute for Water Research (NIVA), Section of Ecotoxicology and Risk Assessment, Gaustadalléen 21, N-0349, Oslo (Norway); Centre for Environmental Radioactivity, Norwegian University of Life Sciences (NMBU), Post Box 5003, N-1432 Ås (Norway); Brede, Dag; Lind, Ole-Christian [Centre for Environmental Radioactivity, Norwegian University of Life Sciences (NMBU), Post Box 5003, N-1432 Ås (Norway); Department for Environmental Sciences, Faculty of Environmental Science & Technology, Norwegian University of Life Sciences (NMBU), Post Box 5003, N-1432, Ås (Norway); Solhaug, Knut Asbjørn [Centre for Environmental Radioactivity, Norwegian University of Life Sciences (NMBU), Post Box 5003, N-1432 Ås (Norway); Department of Ecology and Natural Resource Management, Norwegian University of Life Sciences (NMBU), Postbox 5003, N-1432, Ås (Norway); Salbu, Brit [Centre for Environmental Radioactivity, Norwegian University of Life Sciences (NMBU), Post Box 5003, N-1432 Ås (Norway); Department for Environmental Sciences, Faculty of Environmental Science & Technology, Norwegian University of Life Sciences (NMBU), Post Box 5003, N-1432, Ås (Norway); and others

2017-02-15

Highlights: • Chlorophyll fluorescence parameters affected at higher dose rates. • Changes in PSII associated with electron transport and energy dissipation pathways. • Dose-dependent ROS production in algae exposed to gamma radiation. • Decrease in photosynthetic efficiency connected to ROS formation. - Abstract: The aquatic environment is continuously exposed to ionizing radiation from both natural and anthropogenic sources, making the characterization of ecological and health risks associated with radiation of large importance. Microalgae represent the main source of biomass production in the aquatic ecosystem, thus becoming a highly relevant biological model to assess the impacts of gamma radiation. However, little information is available on the effects of gamma radiation on microalgal species, making environmental radioprotection of this group of species challenging. In this context, the present study aimed to improve the understanding of the effects and toxic mechanisms of gamma radiation in the unicellular green algae Chlamydomonas reinhardtii focusing on the activity of the photosynthetic apparatus and ROS formation. Algal cells were exposed to gamma radiation (0.49–1677 mGy/h) for 6 h and chlorophyll fluorescence parameters obtained by PAM fluorometry, while two fluorescent probes carboxy-H{sub 2}DFFDA and DHR 123 were used for the quantification of ROS. The alterations seen in functional parameters of C. reinhardtii PSII after 6 h of exposure to gamma radiation showed modifications of PSII energy transfer associated with electron transport and energy dissipation pathways, especially at the higher dose rates used. Results also showed that gamma radiation induced ROS in a dose-dependent manner under both light and dark conditions. The observed decrease in photosynthetic efficiency seems to be connected to the formation of ROS and can potentially lead to oxidative stress and cellular damage in chloroplasts. To our knowledge, this is the first

Rapid and Highly Sensitive Non-Competitive Immunoassay for Specific Detection of Nodularin

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Sultana Akter

2017-09-01

Full Text Available Nodularin (NOD is a cyclic penta-peptide hepatotoxin mainly produced by Nodularia spumigena, reported from the brackish water bodies of various parts of the world. It can accumulate in the food chain and, for safety reasons, levels of NOD not only in water bodies but also in food matrices are of interest. Here, we report on a non-competitive immunoassay for the specific detection of NOD. A phage display technique was utilized to interrogate a synthetic antibody phage library for binders recognizing NOD bound to an anti-ADDA (3-Amino-9-methoxy-2,6,8-trimethyl-10-phenyldeca-4(E,6(E-dienoic acid monoclonal antibody (Mab. One of the obtained immunocomplex binders, designated SA32C11, showed very high specificity towards nodularin-R (NOD-R over to the tested 10 different microcystins (microcystin-LR, -dmLR, -RR, -dmRR, -YR, -LY, -LF, -LW, -LA, -WR. It was expressed in Escherichia coli as a single chain antibody fragment (scFv fusion protein and used to establish a time-resolved fluorometry-based assay in combination with the anti-ADDA Mab. The detection limit (blank + 3SD of the immunoassay, with a total assay time of 1 h 10 min, is 0.03 µg/L of NOD-R. This represents the most sensitive immunoassay method for the specific detection of NOD reported so far. The assay was tested for its performance to detect NOD using spiked (0.1 to 3 µg/L of NOD-R water samples including brackish sea and coastal water and the recovery ranged from 79 to 127%. Furthermore, a panel of environmental samples, including water from different sources, fish and other marine tissue specimens, were analyzed for NOD using the assay. The assay has potential as a rapid screening tool for the analysis of a large number of water samples for the presence of NOD. It can also find applications in the analysis of the bioaccumulation of NOD in marine organisms and in the food chain.

Anthropogenic sediments and soils of tells of the Balkans and Anatolia: Composition, genesis, and relationships with the history of landscape and human occupation

Science.gov (United States)

Sedov, S. N.; Aleksandrovskii, A. L.; Benz, M.; Balabina, V. I.; Mishina, T. N.; Shishkov, V. A.; Şahin, F.; Özkaya, V.

2017-04-01

Soils and sediments composing Tell Körtik Tepe (Epipaleolithic, Turkey) and Tell Yunatsite (Chalcolithic (Eneolithic), Bulgaria) have been studied with the aim to gain a better insight into their microfabrics, determine the composition of anthropogenic artifacts, and, on this basis, to analyze similarities and distinctions between these objects and the modern soils of urban areas. The methods of micromorphology, scanning electron microscopy with an energy dispersive X-ray microanalyzer, X-ray fluorometry, and other techniques to determine the chemical and physical properties of the soils and sediments have been applied. Two paleosols have been identified in Tell Yunatsite with a total thickness of 9 m: the paleosol buried under the tell and the paleosol in its middle part. Sediments of Tell Körtik Tepe have a total thickness of up to 5 m; their accumulation began at the end of Pleistocene over the surface of buried paleosol. The cultural layer of the tells consists of construction debris mainly represented by a mixture of clay and sand and of domestic wastes with the high content of phosphorus. The major source of phosphorus is calcium phosphate (apatite) of bone tissues. The abundance of various anthropogenic materials in the sediments is clearly seen in thin sections. Even in the paleosols developed within the cultural layer (the mid-profile paleosol in Tell Yunatsite), the amount of microinclusions of bone fragments, charcoal, and burnt clay (ceramics) is very high. Micromorphological data indicate that up to 50% of the layered material filling an Epipaleolithic construction in Tell Körtik Tepe consists of the anthropogenic inclusions: bone fragments, charcoal, etc. The features of pedogenic transformation are present in the sediments. Such sediments can be classified as synlithogenic soils similar to the modern Urbic Technosols. It is shown that the formation of paleosols and sediments of Tell Körtik Tepe took place under extreme environmental conditions�

Assessing bio-available silver released from silver nanoparticles embedded in silica layers using the green algae Chlamydomonas reinhardtii as bio-sensors

Energy Technology Data Exchange (ETDEWEB)

Pugliara, Alessandro [nMat group-CEMES (Centre d' Elaboration de Matériaux et d' Etudes Structurales)-CNRS, Université de Toulouse, 29 rue Jeanne Marvig, BP 94347, F-31055 Toulouse Cedex 4 (France); LAPLACE (LAboratoire PLAsma et Conversion d' Energie), Université de Toulouse, CNRS, UPS, INPT, 118 route de Narbonne, F-31062 Toulouse (France); Makasheva, Kremena; Despax, Bernard [LAPLACE (LAboratoire PLAsma et Conversion d' Energie), Université de Toulouse, CNRS, UPS, INPT, 118 route de Narbonne, F-31062 Toulouse (France); Bayle, Maxime; Carles, Robert; Benzo, Patrizio; BenAssayag, Gérard; Pécassou, Béatrice [nMat group-CEMES (Centre d' Elaboration de Matériaux et d' Etudes Structurales)-CNRS, Université de Toulouse, 29 rue Jeanne Marvig, BP 94347, F-31055 Toulouse Cedex 4 (France); Sancho, Maria Carmen; Navarro, Enrique [IPE (Instituto Pirenaico de Ecología)-CSIC, Avda. Montañana 1005, Zaragoza 50059 (Spain); Echegoyen, Yolanda [I3A, Department of Analytical Chemistry, University of Zaragoza, C/ María de Luna 3, 50018, Zaragoza (Spain); Bonafos, Caroline, E-mail: bonafos@cemes.fr [nMat group-CEMES (Centre d' Elaboration de Matériaux et d' Etudes Structurales)-CNRS, Université de Toulouse, 29 rue Jeanne Marvig, BP 94347, F-31055 Toulouse Cedex 4 (France)

2016-09-15

Silver nanoparticles (AgNPs) because of their strong antibacterial activity are widely used in health-care sector and industrial applications. Their huge surface-volume ratio enhances the silver release compared to the bulk material, leading to an increased toxicity for microorganisms sensitive to this element. This work presents an assessment of the toxic effect on algal photosynthesis due to small (size < 20 nm) AgNPs embedded in silica layers. Two physical approaches were originally used to elaborate the nanocomposite structures: (i) low energy ion beam synthesis and (ii) combined silver sputtering and plasma polymerization. These techniques allow elaboration of a single layer of AgNPs embedded in silica films at defined nanometer distances (from 0 to 7 nm) beneath the free surface. The structural and optical properties of the nanostructures were studied by transmission electron microscopy and optical reflectance. The silver release from the nanostructures after 20 h of immersion in buffered water was measured by inductively coupled plasma mass spectrometry and ranges between 0.02 and 0.49 μM. The short-term toxicity of Ag to photosynthesis of Chlamydomonas reinhardtii was assessed by fluorometry. The obtained results show that embedding AgNPs reduces the interactions with the buffered water free media, protecting the AgNPs from fast oxidation. The release of bio-available silver (impacting on the algal photosynthesis) is controlled by the depth at which AgNPs are located for a given host matrix. This provides a procedure to tailor the toxicity of nanocomposites containing AgNPs. - Highlights: • Controlled synthesis of 2D arrays of silver nanoparticles embedded in silica. • Assessing bio-available silver release using the green algae as bio-sensors. • The Ag release can be controlled by the distance nanoparticles/dielectric surface. • All the Ag released in solution is in the form of Ag{sup +} ions. • Toxicity comparable to similar concentrations of

Recent Invasion of the Symbiont-Bearing Foraminifera Pararotalia into the Eastern Mediterranean Facilitated by the Ongoing Warming Trend.

Science.gov (United States)

Schmidt, Christiane; Morard, Raphael; Almogi-Labin, Ahuva; Weinmann, Anna E; Titelboim, Danna; Abramovich, Sigal; Kucera, Michal

2015-01-01

The eastern Mediterranean is a hotspot of biological invasions. Numerous species of Indo-pacific origin have colonized the Mediterranean in recent times, including tropical symbiont-bearing foraminifera. Among these is the species Pararotalia calcariformata. Unlike other invasive foraminifera, this species was discovered only two decades ago and is restricted to the eastern Mediterranean coast. Combining ecological, genetic and physiological observations, we attempt to explain the recent invasion of this species in the Mediterranean Sea. Using morphological and genetic data, we confirm the species attribution to P. calcariformata McCulloch 1977 and identify its symbionts as a consortium of diatom species dominated by Minutocellus polymorphus. We document photosynthetic activity of its endosymbionts using Pulse Amplitude Modulated Fluorometry and test the effects of elevated temperatures on growth rates of asexual offspring. The culturing of asexual offspring for 120 days shows a 30-day period of rapid growth followed by a period of slower growth. A subsequent 48-day temperature sensitivity experiment indicates a similar developmental pathway and high growth rate at 28°C, whereas an almost complete inhibition of growth was observed at 20°C and 35°C. This indicates that the offspring of this species may have lower tolerance to cold temperatures than what would be expected for species native to the Mediterranean. We expand this hypothesis by applying a Species Distribution Model (SDM) based on modern occurrences in the Mediterranean using three environmental variables: irradiance, turbidity and yearly minimum temperature. The model reproduces the observed restricted distribution and indicates that the range of the species will drastically expand westwards under future global change scenarios. We conclude that P. calcariformata established a population in the Levant because of the recent warming in the region. In line with observations from other groups of organisms

Recent Invasion of the Symbiont-Bearing Foraminifera Pararotalia into the Eastern Mediterranean Facilitated by the Ongoing Warming Trend.

Directory of Open Access Journals (Sweden)

Christiane Schmidt

Full Text Available The eastern Mediterranean is a hotspot of biological invasions. Numerous species of Indo-pacific origin have colonized the Mediterranean in recent times, including tropical symbiont-bearing foraminifera. Among these is the species Pararotalia calcariformata. Unlike other invasive foraminifera, this species was discovered only two decades ago and is restricted to the eastern Mediterranean coast. Combining ecological, genetic and physiological observations, we attempt to explain the recent invasion of this species in the Mediterranean Sea. Using morphological and genetic data, we confirm the species attribution to P. calcariformata McCulloch 1977 and identify its symbionts as a consortium of diatom species dominated by Minutocellus polymorphus. We document photosynthetic activity of its endosymbionts using Pulse Amplitude Modulated Fluorometry and test the effects of elevated temperatures on growth rates of asexual offspring. The culturing of asexual offspring for 120 days shows a 30-day period of rapid growth followed by a period of slower growth. A subsequent 48-day temperature sensitivity experiment indicates a similar developmental pathway and high growth rate at 28°C, whereas an almost complete inhibition of growth was observed at 20°C and 35°C. This indicates that the offspring of this species may have lower tolerance to cold temperatures than what would be expected for species native to the Mediterranean. We expand this hypothesis by applying a Species Distribution Model (SDM based on modern occurrences in the Mediterranean using three environmental variables: irradiance, turbidity and yearly minimum temperature. The model reproduces the observed restricted distribution and indicates that the range of the species will drastically expand westwards under future global change scenarios. We conclude that P. calcariformata established a population in the Levant because of the recent warming in the region. In line with observations from other

Uranium hydrogeochemical and stream sediment reconnaissance data from the area of the Noatak and portions of the Baird Mountains and Ambler River Quadrangles, Alaska

International Nuclear Information System (INIS)

Aamodt, P.L.; Hill, D.E.; Sharp, R.R. Jr.

1978-05-01

During August 1976, a total of 876 natural waters and 861 bottom sediments were collected at a nominal density of one location each 23 km 2 from streams and small lakes throughout the Noatak NTMS quadrangle, the southern two-thirds of the Baird Mountains NTMS quadrangle, and in the southwest corner of the Ambler River NTMS quadrangle. These samples were collected as part of the National Uranium Resource Evaluation program in Alaska being conducted by the Los Alamos Scientific Laboratory (LASL). The field collection and treatment of the samples were performed following strict LASL specifications. Total uranium was measured in the waters by fluorometry and in the sediments by delayed-neutron counting, using stringent quality assurance controls at the LASL. The uranium contents of the waters ranged from below the detection limit of 0.02 parts per billion (ppB) to a high of 8.38 ppB, and the uranium contents of the sediments ranged from a low of 0.3 parts per million (ppM) to a high of 34.0 ppM. In general, the locations of waters containing relatively high uranium contents were found to occur in clusters, and particularly in the headwaters of streams draining the southern slopes of the Baird Mountains. Few sediments contained relatively high uranium contents. These usually occurred singly at isolated locations scattered throughout the area. No obvious association exists between the location of high-uranium waters and sediments anywhere in the study area. The geology, mineralogy, and hydrology of this area is only generally described in the literature; therefore, it is difficult to correlate these data with particular aspects of the physical environment where individual samples were collected. However, the data do indicate that certain areas underlaid by Paleozoic sedimentary rocks and granitic intrusives within the Baird Mountains and a quartz-pebble conglomerate in the Waring Mountains may warrant more detailed field investigations

Effects of different algaecides on the photosynthetic capacity, cell integrity and microcystin-LR release of Microcystis aeruginosa

Energy Technology Data Exchange (ETDEWEB)

Zhou, Shiqing [State Key Laboratory of Pollution Control and Resource Reuse, Tongji University, Shanghai 200092 (China); Shao, Yisheng, E-mail: yishengshao@163.com [State Key Laboratory of Pollution Control and Resource Reuse, Tongji University, Shanghai 200092 (China); China Academy of Urban Planning and Design, Beijing 100037 (China); Gao, Naiyun [State Key Laboratory of Pollution Control and Resource Reuse, Tongji University, Shanghai 200092 (China); Deng, Yang [Department of Earth and Environmental Studies, Montclair State University, Montclair NJ 07043 (United States); Qiao, Junlian; Ou, Huase; Deng, Jing [State Key Laboratory of Pollution Control and Resource Reuse, Tongji University, Shanghai 200092 (China)

2013-10-01

Bench scale tests were conducted to study the effects of four common algaecides, including copper sulfate, hydrogen peroxide, diuron and ethyl 2-methylacetoacetate (EMA) on the photosynthetic capacity, cell integrity and microcystin-LR (MC-LR) release of Microcystis aeruginosa. The release of potassium (K{sup +}) from cell membrane during algaecide exposure was also analyzed. The three typical photosynthetic parameters, including the effective quantum yield (φ{sub e}), photosynthetic efficiency (α) and maximal electron transport rate (rETR{sub max}), were measured by a pulse amplitude modulated (PAM) fluorometry. Results showed that the photosynthetic capacity was all inhibited by the four algaecides, to different degrees, by limiting the energy capture in photosynthesis, and blocking the electron transfer chain in primary reaction. For example, at high diuron concentration (7.5 mg L{sup −1}), φ{sub e}, α and rETR{sub max} decreased from 0.46 to 0.19 (p < 0.01), from 0.20 to 0.01 (p < 0.01) μmol electrons m{sup −2} s{sup −1}/μmol photons m{sup −2} s{sup −1}, and from 160.7 to 0.1 (p < 0.001) μmol m{sup −2} s{sup −1} compared with the control group after 96 h of exposure, respectively. Furthermore, the increase of algaecide dose could lead to the cell lysis, as well as release of intracellular MC-LR that enhanced the accumulation of extracellular MC-LR. The order of MC-LR release potential for the four algaecides was CuSO{sub 4} > H{sub 2}O{sub 2} > diuron > EMA. Highlights: • PAM was used to investigate the effects of algaecides on Microcystis aeruginosa. • We estimate the release of potassium (K{sup +}) from cell membrane for cell lysis. • The risk of microcystin-LR release was evaluated after algaecides exposure. • The order of MC-LR release potential was copper sulfate > hydrogen peroxide > diuron > ethyl 2-methylacetoacetate.

Uranium Hydrogeochemical and Stream Sediment Reconnaissance data from the area of the Teller, Bendeleben, Candle, and Kateel River Quadrangles, Seward Peninsula and vicinity, Alaska

International Nuclear Information System (INIS)

Sharp, R.R. Jr.; Hill, D.E.

1978-05-01

During July-August 1976, 2026 natural waters and 2085 bottom sediments were collected from 2209 sample locations (at a nominal density of one location each 23 km 2 ) on streams and small lakes throughout the Teller, Bendeleben, Candle, and western one-third of the Kateel River NTMS quadrangles, Alaska. Total uranium was measured in the waters by fluorometry and in the sediments and a few waters by delayed-neutron counting. The uranium content of the waters ranged from below the detection limit of 0.02 parts per billion (ppB) to a high of 14.50 ppB, averaging 0.44 ppB, and that of the sediments ranged from a low of 0.2 parts per million (ppM) to a high of 107.4 ppM, averaing 3.93 ppM. The uranium data for water and sediment are separately presented--as computer listings that include pertinent field measurements from each location, as graphically portrayed concentration overlays at 1:250,000 scale for each quadrangle, and as reduced figures showing contours drawn at various concentration levels for each quadrangle--and their areal distributions are compared and correlated with the known features and uranium showings. A test of increasingly detailed methods of data evaluation shows that the more extensive the evaluation, the more useful the reconnaissance uranium data are likely to be. The validity and potential usefulness of the HSSR uranium data are conclusively substantiated by the fact that evidence of all 23 of the reported uranium showings in the 50,000-km 2 study area can be discerned. Several new locations of interest for further field investigation are identified in each of the quadrangles, and most notably in the Bendeleben Mountains. However, the data presented would appear equally useful in guiding field investigation around the uranium occurrences already known, as noteworthy samples often come from close by but on tributary drainages adjacent, opposite, or above them

Apomorphine and piribedil in rats: biochemical and pharmacologic studies.

Science.gov (United States)

Butterworth, R F; Poignant, J C; Barbeau, A

1975-01-01

We studied the biochemical and pharmacologic modes of action of piribedil and apomorphine in the rat. Although both drugs have many points in common, they are also different in many of their manifestations. Apomorphine causes high-intensity, short-duration stereotyped behavior; it is distributed within the brain in uneven fashion, the striatum being the area of lowest concentration as measured by fluorometry. Direct stereotactic injection within the dopaminergic mesolimbic system, and particularly the tuberculum olfactorium, produced constant intense responses. All effects of apomorphine can be blocked by pimozide, but propanolol, a beta blocker, only reduces aggression and ferocity, leaving stereotyped behaviors intact. Finally, L-5-HTP tends to reduce aggression, ferocity, and to a lesser extent stereotypy; MIF or piribedil, as well as reserpine, potentiates the stereotyped behaviors induced by apomorphine, whereas L-DOPA usually decreases them. Piribedil, on the other hand, causes low-intensity, long-duration stereotyped behavior. It is distributed within the brain almost uniformly. Most effects of piribedil can be blocked by pimozide, but propanolol blocks only aggression and ferocity, leaving stereotyped behaviors intact. On the other hand, clonidine, an alpha-receptor agonist, blocks stereotyped behaviors induced by piribedil but markedly increases aggression, ferocity, and motor activity. L-5-HTP and L-DOPA have little effect on piribedil-induced manifestations. Reserpine decreases piribedil stereotypy. The main metabolite of piribedil, S 584, had no clear-cut pharmacologic action in our hands at the dosage used. It is concluded that both apomorphine and piribedil produce stereotyped behavior by modifying the physiologic balance between mesolimbic and nigrostriatal dopaminergic systems. The other actions of apomorphine and piribedil upon aggression, ferocity, and motor activity are not always in parallel and depend probably on the fact that piribedil is less

A fluorescence-based method for direct measurement of submicrosecond intramolecular contact formation in biopolymers: an exploratory study with polypeptides.

Science.gov (United States)

Hudgins, Robert R; Huang, Fang; Gramlich, Gabriela; Nau, Werner M

2002-01-30

A fluorescent amino acid derivative (Fmoc-DBO) has been synthesized, which contains 2,3-diazabicyclo[2.2.2]oct-2-ene (DBO) as a small, hydrophilic fluorophore with an extremely long fluorescence lifetime (325 ns in H2O and 505 ns in D2O under air). Polypeptides containing both the DBO residue and an efficient fluorescence quencher allow the measurement of rate constants for intramolecular end-to-end contact formation. Bimolecular quenching experiments indicated that Trp, Cys, Met, and Tyr are efficient quenchers of DBO (k(q) = 20, 5.1, 4.5, and 3.6 x 10(8) M(-1) x s(-1) in D2O), while the other amino acids are inefficient. The quenching by Trp, which was selected as an intrinsic quencher, is presumed to involve exciplex-induced deactivation. Flexible, structureless polypeptides, Trp-(Gly-Ser)n-DBO-NH2, were prepared by standard solid-phase synthesis, and the rates of contact formation were measured through the intramolecular fluorescence quenching of DBO by Trp with time-correlated single-photon counting, laser flash photolysis, and steady-state fluorometry. Rate constants of 4.1, 6.8, 4.9, 3.1, 2.0, and 1.1 x 10(7) s(-1) for n = 0, 1, 2, 4, 6, and 10 were obtained. Noteworthy was the relatively slow quenching for the shortest peptide (n = 0). The kinetic data are in agreement with recent transient absorption studies of triplet probes for related peptides, but the rate constants are significantly larger. In contrast to the flexible structureless Gly-Ser polypeptides, the polyproline Trp-Pro4-DBO-NH2 showed insignificant fluorescence quenching, suggesting that a high polypeptide flexibility and the possibility of probe-quencher contact is essential to induce quenching. Advantages of the new fluorescence-based method for measuring contact formation rates in biopolymers include high accuracy, fast time range (100 ps-1 micros), and the possibility to perform measurements in water under air.

Sensitivity of the green algae Chlamydomonas reinhardtii to gamma radiation: Photosynthetic performance and ROS formation

International Nuclear Information System (INIS)

Gomes, Tânia; Xie, Li; Brede, Dag; Lind, Ole-Christian; Solhaug, Knut Asbjørn; Salbu, Brit

2017-01-01

Highlights: • Chlorophyll fluorescence parameters affected at higher dose rates. • Changes in PSII associated with electron transport and energy dissipation pathways. • Dose-dependent ROS production in algae exposed to gamma radiation. • Decrease in photosynthetic efficiency connected to ROS formation. - Abstract: The aquatic environment is continuously exposed to ionizing radiation from both natural and anthropogenic sources, making the characterization of ecological and health risks associated with radiation of large importance. Microalgae represent the main source of biomass production in the aquatic ecosystem, thus becoming a highly relevant biological model to assess the impacts of gamma radiation. However, little information is available on the effects of gamma radiation on microalgal species, making environmental radioprotection of this group of species challenging. In this context, the present study aimed to improve the understanding of the effects and toxic mechanisms of gamma radiation in the unicellular green algae Chlamydomonas reinhardtii focusing on the activity of the photosynthetic apparatus and ROS formation. Algal cells were exposed to gamma radiation (0.49–1677 mGy/h) for 6 h and chlorophyll fluorescence parameters obtained by PAM fluorometry, while two fluorescent probes carboxy-H 2 DFFDA and DHR 123 were used for the quantification of ROS. The alterations seen in functional parameters of C. reinhardtii PSII after 6 h of exposure to gamma radiation showed modifications of PSII energy transfer associated with electron transport and energy dissipation pathways, especially at the higher dose rates used. Results also showed that gamma radiation induced ROS in a dose-dependent manner under both light and dark conditions. The observed decrease in photosynthetic efficiency seems to be connected to the formation of ROS and can potentially lead to oxidative stress and cellular damage in chloroplasts. To our knowledge, this is the first report

Fabrication and characterization of MCC approved testing material - ATM-1 glass

International Nuclear Information System (INIS)

Wald, J.W.

1985-10-01

The Materials Characterization Center Approved Testing Material ATM-1 is a borosilicate glass that incorporates nonradioactive constituents and uranium to represent high-level waste (HLW) resulting from the reprocessing of commercial nuclear reactor fuel. Its composition is based upon the simulated HLW glass type 76-68 to which depleted uranium has been added as UO 2 . Three separate lots of ATM-1 glass have been fabricated, designated ATM-1a, ATM-1b, and ATM-1c. Limited analyses and microstructural evaluations were conducted on each type. Each lot of ATM-1 glass was produced from a feedstock melted in an air atmosphere at between 1150 to 1200 0 C and cast into stress annealed rectangular bars. Bars of ATM-1a were nominally 1.3 x 1.3 x 7.6 cm (approx.36 g each), bars of ATM-1b were nominally 2 x 2.5 x 17.5 cm (approx.190 g each) and bars of ATM-1c were nominally 1.9 x 1.9 x 15 cm (approx.170 g each). Thirteen bars of ATM-1a, 14 bars of ATM-1b, and 6 bars of ATM-1c were produced. Twelve random samples from each of lots ATM-1a, ATM-1b, and ATM-1c were analyzed. The concentrations (except for U and Cs) were obtained by Inductively-Coupled Argon Plasma Atomic Emission Spectroscopy analysis. Cesium analysis was performed by Atomic Absorption Spectroscopy, while uranium was analyzed by Pulsed Laser Fluorometry. X-ray diffraction analysis of four samples indicated that lot ATM-1a had no detectable crystalline phases (<3 wt %), while ATM-1b and ATM-1c contained approx.3 to 5 wt % iron-chrome spinel crystals. These concentrations of secondary spinel component are not considered uncommon. Scanning electron microscopy examination of fracture surfaces revealed only a random, apparently crystalline, second phase (1-10 μm diam) and a random distribution of small voids or bubbles (approx.1 μm nominal diam)

Potential role of sodium-proton exchangers in the low concentration arsenic trioxide-increased intracellular pH and cell proliferation.

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Carmen Aravena

Full Text Available Arsenic main inorganic compound is arsenic trioxide (ATO presented in solution mainly as arsenite. ATO increases intracellular pH (pHi, cell proliferation and tumor growth. Sodium-proton exchangers (NHEs modulate the pHi, with NHE1 playing significant roles. Whether ATO-increased cell proliferation results from altered NHEs expression and activity is unknown. We hypothesize that ATO increases cell proliferation by altering pHi due to increased NHEs-like transport activity. Madin-Darby canine kidney (MDCK cells grown in 5 mmol/L D-glucose-containing DMEM were exposed to ATO (0.05, 0.5 or 5 µmol/L, 0-48 hours in the absence or presence of 5-N,N-hexamethylene amiloride (HMA, 5-100 µmol/L, NHEs inhibitor, PD-98059 (30 µmol/L, MAPK1/2 inhibitor, Gö6976 (10 µmol/L, PKCα, βI and μ inhibitor, or Schering 28080 (10 µmol/L, H(+/K(+ATPase inhibitor plus concanamycin (0.1 µmol/L, V type ATPases inhibitor. Incorporation of [(3H]thymidine was used to estimate cell proliferation, and counting cells with a hemocytometer to determine the cell number. The pHi was measured by fluorometry in 2,7-bicarboxyethyl-5,6-carboxyfluorescein loaded cells. The Na(+-dependent HMA-sensitive NHEs-like mediated proton transport kinetics, NHE1 protein abundance in the total, cytoplasm and plasma membrane protein fractions, and phosphorylated and total p42/44 mitogen-activated protein kinases (p42/44(mapk were also determined. Lowest ATO (0.05 µmol/L, ~0.01 ppm used in this study increased cell proliferation, pHi, NHEs-like transport and plasma membrane NHE1 protein abundance, effects blocked by HMA, PD-98059 or Gö6976. Cell-buffering capacity did not change by ATO. The results show that a low ATO concentration increases MDCK cells proliferation by NHEs (probably NHE1-like transport dependent-increased pHi requiring p42/44(mapk and PKCα, βI and/or μ activity. This finding could be crucial in diseases where uncontrolled cell growth occurs, such as tumor growth, and

Utility of Photochemical Traits as Diagnostics of Thermal Tolerance amongst Great Barrier Reef Corals

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Matthew R. Nitschke

2018-02-01

Full Text Available Light availability is considered a key factor regulating the thermal sensitivity of reef building corals, where excessive excitation of photosystem II (PSII further exacerbates pressure on photochemical pathways already compromised by heat stress. Coral symbionts acclimate to changes in light availability (photoacclimation by continually fine-tuning the photochemical operating efficiency of PSII. However, how this process adjusts throughout the warmest months in naturally heat-tolerant or sensitive species is unknown, and whether this influences the capacity to tolerate transient heat stress is untested. We therefore examined the PSII photophysiology of 10 coral species (with known thermal tolerances from shallow reef environments at Heron Island (Great Barrier Reef, Australia, in spring (October-November, 2015 vs. summer (February-March, 2016. Corals were maintained in flow-through aquaria and rapid light curve (RLC protocols using pulse amplitude modulated (PAM fluorometry captured changes in the PSII photoacclimation strategy, characterized as the minimum saturating irradiance (Ek, and the extent of photochemical ([1 – C], operating efficiency vs. non-photochemical ([1 – Q] energy dissipation. Values of Ek across species were >2-fold higher in all coral species in spring, consistent with a climate of higher overall light exposure (i.e., higher PAR from lower cloud cover, rainfall and wind speed compared with summer. Summer decreases in Ek were combined with a shift toward preferential photochemical quenching in all species. All coral species were subsequently subjected to thermal stress assays. An equivalent temperature-ramping profile of 1°C increase per day and then maintenance at 32°C was applied in each season. Despite the significant seasonal photoacclimation, the species hierarchy of thermal tolerance [maximum quantum yields of PSII (Fv/Fm, monitored at dawn and dusk] did not shift between seasons, except for Pocillopora

Change in algal symbiont communities after bleaching, not prior heat exposure, increases heat tolerance of reef corals.

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Silverstein, Rachel N; Cunning, Ross; Baker, Andrew C

2015-01-01

Mutualistic organisms can be particularly susceptible to climate change stress, as their survivorship is often limited by the most vulnerable partner. However, symbiotic plasticity can also help organisms in changing environments by expanding their realized niche space. Coral-algal (Symbiodinium spp.) symbiosis exemplifies this dichotomy: the partnership is highly susceptible to 'bleaching' (stress-induced symbiosis breakdown), but stress-tolerant symbionts can also sometimes mitigate bleaching. Here, we investigate the role of diverse and mutable symbiotic partnerships in increasing corals' ability to thrive in high temperature conditions. We conducted repeat bleaching and recovery experiments on the coral Montastraea cavernosa, and used quantitative PCR and chlorophyll fluorometry to assess the structure and function of Symbiodinium communities within coral hosts. During an initial heat exposure (32 °C for 10 days), corals hosting only stress-sensitive symbionts (Symbiodinium C3) bleached, but recovered (at either 24 °C or 29 °C) with predominantly (>90%) stress-tolerant symbionts (Symbiodinium D1a), which were not detected before bleaching (either due to absence or extreme low abundance). When a second heat stress (also 32 °C for 10 days) was applied 3 months later, corals that previously bleached and were now dominated by D1a Symbiodinium experienced less photodamage and symbiont loss compared to control corals that had not been previously bleached, and were therefore still dominated by Symbiodinium C3. Additional corals that were initially bleached without heat by a herbicide (DCMU, at 24 °C) also recovered predominantly with D1a symbionts, and similarly lost fewer symbionts during subsequent thermal stress. Increased thermotolerance was also not observed in C3-dominated corals that were acclimated for 3 months to warmer temperatures (29 °C) before heat stress. These findings indicate that increased thermotolerance post-bleaching resulted from

Voltage-sensing phosphatase modulation by a C2 domain

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Paul M. Castle

2015-04-01

Full Text Available The voltage-sensing phosphatase (VSP is the first example of an enzyme controlled by changes in membrane potential. VSP has four distinct regions: the transmembrane voltage-sensing domain (VSD, the inter-domain linker, the cytosolic catalytic domain and the C2 domain. The VSD transmits the changes in membrane potential through the inter-domain linker activating the catalytic domain which then dephosphorylates phosphatidylinositol phosphate lipids. The role of the C2, however, has not been established. In this study, we explore two possible roles for the C2: catalysis and membrane-binding. The Ci-VSP crystal structures show that the C2 residue Y522 lines the active site suggesting a contribution to catalysis. When we mutated Y522 to phenylalanine, we found a shift in the voltage dependence of activity. This suggests hydrogen bonding as a mechanism of action. Going one step further, when we deleted the entire C2 domain, we found voltage-dependent enzyme activity was no longer detectable. This result clearly indicates the entire C2 is necessary for catalysis as well as for modulating activity. As C2s are known membrane-binding domains, we tested whether the VSP C2 interacts with the membrane. We probed a cluster of four positively charged residues lining the top of the C2 and suggested by previous studies to interact with phosphatidylinositol 4,5-bisphosphate (PI(4,5P2 (Kalli et al., 2014. Neutralizing those positive charges significantly shifted the voltage dependence of activity to higher voltages. We tested membrane binding by depleting PI(4,5P2 from the membrane using the 5HT2C receptor and found that the VSD motions as measured by voltage clamp fluorometry were not changed. These results suggest that if the C2 domain interacts with the membrane to influence VSP function it may not occur exclusively through PI(4,5P2. Together, this data advances our understanding of the VSP C2 by demonstrating a necessary and critical role for the C2 domain in

Epidemiologic and Molecular Relationship Between Vaccine Manufacture and Autism Spectrum Disorder Prevalence.

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Deisher, Theresa A; Doan, Ngoc V; Koyama, Kumiko; Bwabye, Sarah

2015-01-01

To assess the public health consequences of fetal cell line manufactured vaccines that contain residual human fetal DNA fragments utilizing laboratory and ecological approaches including statistics, molecular biology and genomics. MMR coverage and autism disorder or autism spectrum disorder prevalence data for Norway, Sweden and the UK were obtained from public and government websites as well as peer reviewed published articles. Biologically, the size and quantity of the contaminating fetal DNA in Meruvax II and Havrix as well as the propensity of various cell lines for cellular and nuclear uptake of primitive human DNA fragments were measured and quantified using gel electrophoresis, fluorescence microscopy and fluorometry. Lastly, genomic analysis identified the specific sites where fetal DNA fragment integration into a child's genome is most likely to occur. The average MMR coverage for the three countries fell below 90% after Dr. Wakefield's infamous 1998 publication but started to recover slowly after 2001 until reaching over 90% coverage again by 2004. During the same time period, the average autism spectrum disorder prevalence in the United Kingdom, Norway and Sweden dropped substantially after birth year 1998 and gradually increased again after birth year 2000. Average single stranded DNA and double stranded DNA in Meruvax II were 142.05 ng/vial and 35.00 ng/vial, respectively, and 276.00 ng/vial and 35.74 ng/vial in Havrix respectively. The size of the fetal DNA fragments in Meruvax II was approximately 215 base pairs. There was spontaneous cellular and nuclear DNA uptake in HFF1 and NCCIT cells. Genes that have been linked to autism (autism associated genes; AAGs) have a more concentrated susceptibility for insults to genomic stability in comparison to the group of all genes contained within the human genome. Of the X chromosome AAGs, 15 of 19 have double strand break motifs less than 100 kilobases away from the center of a meiotic recombination hotspot

Lethal and sub-lethal chronic effects of the herbicide diuron on seagrass.

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Negri, Andrew P; Flores, Florita; Mercurio, Phil; Mueller, Jochen F; Collier, Catherine J

2015-08-01

Photosystem II herbicides from agricultural sources have been detected throughout nearshore tropical habitats including seagrass meadows. While PSII herbicides have been shown to inhibit growth in microalgae at low concentrations, the potential impacts of chronic low concentration exposures to seagrass health and growth have not been investigated. Here we exposed two tropical seagrass species Halodule uninervis and Zostera muelleri to elevated diuron concentrations (from 0.3 to 7.2μgl(-1)) over a 79-day period followed by a 2-week recovery period in uncontaminated seawater. PAM fluorometry demonstrated rapid effect of diuron on photosystem II (PSII) in both seagrass species at 0.3μgl(-1). This effect included significant inhibition of photosynthetic efficiency (ΔF/Fm') and inactivation of PSII (Fv/Fm) over the 11 week exposure period. Significant mortality and reductions in growth was only observed at the highest exposure concentration of 7.2μgl(-1) diuron. However, biochemical indicators demonstrated that the health of seagrass after this prolonged exposure was significantly compromised at lower concentrations. For example, the drop in C:N ratios (0.6μgl(-1)) and reduced δ(13)C (1.7μgl(-1)) in seagrass leaves indicated reduced C-assimilation from photosynthesis. Critically, the energetic reserves of the plants (as measured by starch content in the root-rhizome complex) were approximately halved following diuron exposure at and above 1.7μgl(-1). During the 2-week recovery period, the photosynthetic capacity of the seagrass improved with only plants from the highest diuron treatment still exhibiting chronic damage to PSII. This study shows that, although seagrass may survive prolonged herbicide exposures, concentrations ≥0.6μgl(-1) diuron equivalents cause measureable impacts on energetic status that may leave the plants vulnerable to other simultaneous stressors. For example, tropical seagrasses have been heavily impacted by reduced light from coastal

Critical evaluation of gamma-irradiated serum used as feeder in the culture and demonstration of putative nanobacteria and calcifying nanoparticles.

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Jan Martel

Full Text Available The culture and demonstration of putative nanobacteria (NB and calcifying nanoparticles (CNP from human and animal tissues has relied primarily on the use of a culture supplement consisting of FBS that had been gamma-irradiated at a dose of 30 kGy (gamma-FBS. The use of gamma-FBS is based on the assumption that this sterilized fluid has been rid entirely of any residual NB/CNP, while it continues to promote the slow growth in culture of NB/CNP from human/animal tissues. We show here that gamma-irradiation (5-50 kGy produces extensive dose-dependent serum protein breakdown as demonstrated through UV and visible light spectrophotometry, fluorometry, Fourier-transformed infrared spectroscopy, and gel electrophoresis. Yet, both gamma-FBS and gamma-irradiated human serum (gamma-HS produce NB/CNP in cell culture conditions that are morphologically and chemically indistinguishable from their normal serum counterparts. Contrary to earlier claims, gamma-FBS does not enhance the formation of NB/CNP from several human body fluids (saliva, urine, ascites, and synovial fluid tested. In the presence of additional precipitating ions, both gamma-irradiated serum (FBS and HS and gamma-irradiated proteins (albumin and fetuin-A retain the inherent dual NB inhibitory and seeding capabilities seen also with their untreated counterparts. By gel electrophoresis, the particles formed from both gamma-FBS and gamma-HS are seen to have assimilated into their scaffold the same smeared protein profiles found in the gamma-irradiated sera. However, their protein compositions as identified by proteomics are virtually identical to those seen with particles formed from untreated serum. Moreover, particles derived from human fluids and cultured in the presence of gamma-FBS contain proteins derived from both gamma-FBS and the human fluid under investigation-a confusing and unprecedented scenario indicating that these particles harbor proteins from both the host tissue and the FBS

Gonadotropin-releasing hormone receptor activates GTPase RhoA and inhibits cell invasion in the breast cancer cell line MDA-MB-231

International Nuclear Information System (INIS)

Aguilar-Rojas, Arturo; Huerta-Reyes, Maira; Maya-Núñez, Guadalupe; Arechavaleta-Velásco, Fabián; Conn, P Michael; Ulloa-Aguirre, Alfredo; Valdés, Jesús

2012-01-01

Gonadotropin-releasing hormone (GnRH) and its receptor (GnRHR) are both expressed by a number of malignant tumors, including those of the breast. In the latter, both behave as potent inhibitors of invasion. Nevertheless, the signaling pathways whereby the activated GnRH/GnRHR system exerts this effect have not been clearly established. In this study, we provide experimental evidence that describes components of the mechanism(s) whereby GnRH inhibits breast cancer cell invasion. Actin polymerization and substrate adhesion was measured in the highly invasive cell line, MDA-MB-231 transiently expressing the wild-type or mutant DesK191 GnRHR by fluorometry, flow cytometric analysis, and confocal microscopy, in the absence or presence of GnRH agonist. The effect of RhoA-GTP on stress fiber formation and focal adhesion assembly was measured in MDA-MB-231 cells co-expressing the GnRHRs and the GAP domain of human p190Rho GAP-A or the dominant negative mutant GAP-Y1284D. Cell invasion was determined by the transwell migration assay. Agonist-stimulated activation of the wild-type GnRHR and the highly plasma membrane expressed mutant GnRHR-DesK191 transiently transfected to MDA-MB-231 cells, favored F-actin polymerization and substrate adhesion. Confocal imaging allowed detection of an association between F-actin levels and the increase in stress fibers promoted by exposure to GnRH. Pull-down assays showed that the effects observed on actin cytoskeleton resulted from GnRH-stimulated activation of RhoA GTPase. Activation of this small G protein favored the marked increase in both cell adhesion to Collagen-I and number of focal adhesion complexes leading to inhibition of the invasion capacity of MDA-MB-231 cells as disclosed by assays in Transwell Chambers. We here show that GnRH inhibits invasion of highly invasive breast cancer-derived MDA-MB-231 cells. This effect is mediated through an increase in substrate adhesion promoted by activation of RhoA GTPase and formation of

Cyclopamine tartrate, an inhibitor of Hedgehog signaling, strongly interferes with mitochondrial function and suppresses aerobic respiration in lung cancer cells

International Nuclear Information System (INIS)

Alam, Md Maksudul; Sohoni, Sagar; Kalainayakan, Sarada Preeta; Garrossian, Massoud; Zhang, Li

2016-01-01

Aberrant Hedgehog (Hh) signaling is associated with the development of many cancers including prostate cancer, gastrointestinal cancer, lung cancer, pancreatic cancer, ovarian cancer, and basal cell carcinoma. The Hh signaling pathway has been one of the most intensely investigated targets for cancer therapy, and a number of compounds inhibiting Hh signaling are being tested clinically for treating many cancers. Lung cancer causes more deaths than the next three most common cancers (colon, breast, and prostate) combined. Cyclopamine was the first compound found to inhibit Hh signaling and has been invaluable for understanding the function of Hh signaling in development and cancer. To find novel strategies for combating lung cancer, we decided to characterize the effect of cyclopamine tartrate (CycT), an improved analogue of cyclopamine, on lung cancer cells and its mechanism of action. The effect of CycT on oxygen consumption and proliferation of non-small-cell lung cancer (NSCLC) cell lines was quantified by using an Oxygraph system and live cell counting, respectively. Apoptosis was detected by using Annexin V and Propidium Iodide staining. CycT’s impact on ROS generation, mitochondrial membrane potential, and mitochondrial morphology in NSCLC cells was monitored by using fluorometry and fluorescent microscopy. Western blotting and fluorescent microscopy were used to detect the levels and localization of Hh signaling targets, mitochondrial fission protein Drp1, and heme-related proteins in various NSCLC cells. Our findings identified a novel function of CycT, as well as another Hh inhibitor SANT1, to disrupt mitochondrial function and aerobic respiration. Our results showed that CycT, like glutamine depletion, caused a substantial decrease in oxygen consumption in a number of NSCLC cell lines, suppressed NSCLC cell proliferation, and induced apoptosis. Further, we found that CycT increased ROS generation, mitochondrial membrane hyperpolarization, and

Non-competitive ELISA with broad specificity for microcystins and nodularins

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Sultana Akter

2017-05-01

Full Text Available Simple and cost-effective methods with sufficient sensitivities for preliminary screening of cyanobacterial toxins are in high demand for assessing water quality and safety. We have recently developed a highly sensitive and rapid time-resolved fluorometry based non-competitive immunoassay for detection of microcystins and nodularins. The assay is based on a synthetic broad-specific anti-immunocomplex antibody SA51D1 capable of recognizing the immunocomplex formed by a generic anti-Adda monoclonal antibody (mAb bound to either microcystins or nodularins. Using the same antibody pair, here we describe a very simple and cost-efficient non-competitive ELISA test for microcystins and nodularins based on conventional alkaline phosphatase (AP activity measurement. The recombinant SA51D1 single-chain fragment of antibody variable domain (scFv was produced as a fusion with bacterial alkaline phosphatase in Escherichia coli. After one step affinity purification through His-tag, the scFv-AP fusion protein could directly be used in the assay. For the assay, toxin standard/sample, biotinylated anti-Adda mAb and the scFv-AP were incubated together for one hour on streptavidin-coated microtiter wells, washed and AP activity was then measured by incubating (1 h at 37 ˚C with chromogenic substrate para-nitrophenylphosphate (pNPP. The assay was capable of detecting all the eleven tested toxin variants (microcystin-LR, -dmLR, -RR, -dmRR, -YR, LA -LY, -LF -LW, -WR, and nodularin-R below WHO guide line value of 1 µg L-1. The detection limit (based on blank+3SD response for microcystin-LR was ~0.2 µg L-1. The assay was verified using spiked (0.25 - 4 µg L-1 of microcystin-LR tap, river and lake water samples with recoveries from 64 to 101%. The assay showed good correlation (r2>0.9 with four reference methods for its performance in detecting extracted intracellular microcystin/nodularin from 17 natural surface water samples. The described easy-to-perform assay

Photosynthetic activity, photoprotection and photoinhibition in intertidal microphytobenthos as studied in situ using variable chlorophyll fluorescence

Science.gov (United States)

Serôdio, João; Vieira, Sónia; Cruz, Sónia

2008-06-01

The photosynthetic activity of microphytobenthos biofilms was studied in situ on an intertidal mudflat of the Ria de Aveiro, Portugal. Time series of physical variables characterizing the microenvironment at the sediment photic zone (incident solar irradiance, temperature, salinity), photophysiological parameters and productive biomass of undisturbed microalgal assemblages were measured during daytime low-tide periods along one spring-neap tidal cycle, with the objective of (1) characterizing the short-term variability in photosynthetic activity in situ, (2) relating it with the changing environmental conditions and (3) with the operation of physiologically (xanthophyll cycle) and behaviorally (vertical migration) based photoprotective processes, and (4) assessing the occurrence of photoinhibition. Pulse Amplitude Modulated (PAM) fluorometry was applied to measure photosynthetic activity (the effective and maximum quantum yield of photosystem II, Δ F/ Fm' and Fv/ Fm; the photosynthesis index EFY; rapid light-response curves (RLC)), the photoprotective operation of the xanthophyll cycle and photoinhibition (non-photochemical quenching, NPQ; quantum efficiency of open RCs, Fv'/ Fm'), and vertical migration (productive biomass, Fo). The photosynthetic activity was found to be strongly affected by the cumulative light dose received during the morning low-tide periods. The fluorescence indices Δ F/ Fm', EFY, Fv'/ Fm' and RLC parameters were more depressed under high irradiances when clear sky was present during the morning low tide than when foggy conditions reduced the light dose received during a comparable period. Productive biomass exhibited maximum values in the first hours of the morning, followed by a steep decrease when irradiance reached moderate levels, due to the downward migration of the microalgae. This photophobic migratory response appeared to display a photoprotective role, allowing Δ F/ Fm' to remain near optimum values until irradiance reached

Oxidant-Antioxidant Balance in Severe Brain Injury

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N. N. Yepifantseva

2010-01-01

Full Text Available Objective: to study the time course of changes in oxidative status parameters and their relationship with inflammation mediators in the acute period of severe brain injury (SBI. Subjects and methods. One hundred and thirteen patients aged 17—67 years were examined. The injury was closed and open in 54 (47.8% and 59 (52.2% patients, respectively. Severe brain contusions were observed in 47 patients, diffuse axonal lesions were seen in 2, and intracranial hematomas were present in 64 patients. The Glasgow coma scores for admission consciousness loss were 6.8±0.25. A control group comprised 23 healthy individuals. The significance of differences was estimated by Student’s test, Wilcoxon-Mann-Whitney, test, Spearman’s correlation test. Venous blood samples were used to study total oxidative activity (TOA and total antioxidative activity (TAA, diene conjugates, lactic acid, albumin, transferrin (TF, ceruloplasmin, C-reactive protein, and lactoferrin (LF were measured in venous blood on disease days 1, 4, 7, 10, 14, and 21. The profile of plasma cytokines (IL-1j8, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, IL-12p70, TNF-а, and IFN-y was studied by flow fluorometry on a Cytomics FC 500 cytofluorometer (Beckman Counlter, USA (reagents were from Bender Medsystems, Austria. Results. In SBI, there was an increase in oxidants, a reduction in antioxidant activity, and lipid peroxidation activation, which were closely related. The oxidation coefficient (TOA/TAA was 40 times greater than the normal values on days 7 to 10. The oxidation parameters were found to be associated with inflammation and cytokine-mediated immunological reactions. The time course of changes in the study proteins was characteristic for systemic inflammation and there was an association with oxidative processes only for ceruloplasm. TF was found to have an association with IL-5 and IL-10, which reflects its involvement in immunological reactions. The association with hypoxia was

Caught in a net: Retention efficiency of microplankton ≥ 10 and < 50 μm collected on mesh netting

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

2018-03-01

Living organisms ≥ 10 μm and loss of organisms that, in turn, may underestimate the concentration of organisms within samples. To address this loss, the retention efficiency (RE) was determined for six filtration approaches using laboratory cultures of microalgae and ambient marine organisms. The approaches employed a membrane filter or mesh nettings of different compositions (nylon, stainless steel, polyester, and polycarbonate), nominal pore sizes (5, 7, and 10 μm), and filtering sequences (e.g., pre-filtering water through a coarse filter). Additionally, in trials with polycarbonate track etched (PCTE) membrane filters, water was amended with particulate material to increase turbidity. Organisms ≥ 10 μm were counted in the material retained on the filter (the filtrand), the material passing through the filter (the filtrate), and the whole water (i.e., unfiltered water). In addition, variable fluorescence fluorometry was used to gauge the relative photochemical yield of phytoplankton-a proximal measurement of the physiological status of phytoplankton-in the size fractions. Further, the mesh types and filters were examined using scanning electron microscopy, which showed irregular openings. The RE of cultured organisms-calculated as the concentration in the filtrand relative to combined concentration in the filtrand and the filtrate-was high for all filtration approaches when laboratory cultures were assessed (> 93%), but RE ranged from 66 to 98% when mixed assemblages of ambient organisms were evaluated. Although PCTE membrane filters had the highest RE (98%), it was not significantly higher than the efficiencies of the 7-μm polyester, Double 7-μm polyester, and Dual 35-μm and 7-μm polyester approaches, but it was significantly higher than the 5-μm nylon and 5-μm stainless steel techniques. This result suggests that PCTE membrane filters perform comparably to 7-μm polyester meshes, so that any of these approaches could be used for concentrating

A Bacterial Pathogen Displaying Temperature-Enhanced Virulence of the Microalga Emiliania huxleyi.

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Mayers, Teaghan J; Bramucci, Anna R; Yakimovich, Kurt M; Case, Rebecca J

2016-01-01

Emiliania huxleyi is a globally abundant microalga that plays a significant role in biogeochemical cycles. Over the next century, sea surface temperatures are predicted to increase drastically, which will likely have significant effects on the survival and ecology of E. huxleyi. In a warming ocean, this microalga may become increasingly vulnerable to pathogens, particularly those with temperature-dependent virulence. Ruegeria is a genus of Rhodobacteraceae whose population size tracks that of E. huxleyi throughout the alga's bloom-bust lifecycle. A representative of this genus, Ruegeria sp. R11, is known to cause bleaching disease in a red macroalga at elevated temperatures. To investigate if the pathogenicity of R11 extends to microalgae, it was co-cultured with several cell types of E. huxleyi near the alga's optimum (18°C), and at an elevated temperature (25°C) known to induce virulence in R11. The algal populations were monitored using flow cytometry and pulse-amplitude modulated fluorometry. Cultures of algae without bacteria remained healthy at 18°C, but lower cell counts in control cultures at 25°C indicated some stress at the elevated temperature. Both the C (coccolith-bearing) and S (scale-bearing swarming) cell types of E. huxleyi experienced a rapid decline resulting in apparent death when co-cultured with R11 at 25°C, but had no effect on N (naked) cell type at either temperature. R11 had no initial negative impact on C and S type E. huxleyi population size or health at 18°C, but caused death in older co-cultures. This differential effect of R11 on its host at 18 and 25°C suggest it is a temperature-enhanced opportunistic pathogen of E. huxleyi. We also detected caspase-like activity in dying C type cells co-cultured with R11, which suggests that programmed cell death plays a role in the death of E. huxleyi triggered by R11 - a mechanism induced by viruses (EhVs) and implicated in E. huxleyi bloom collapse. Given that E. huxleyi has recently been

A Bacterial Pathogen Displaying Temperature-Enhanced Virulence of the Microalga Emiliania huxleyi

Science.gov (United States)

Mayers, Teaghan J.; Bramucci, Anna R.; Yakimovich, Kurt M.; Case, Rebecca J.

2016-01-01

Emiliania huxleyi is a globally abundant microalga that plays a significant role in biogeochemical cycles. Over the next century, sea surface temperatures are predicted to increase drastically, which will likely have significant effects on the survival and ecology of E. huxleyi. In a warming ocean, this microalga may become increasingly vulnerable to pathogens, particularly those with temperature-dependent virulence. Ruegeria is a genus of Rhodobacteraceae whose population size tracks that of E. huxleyi throughout the alga’s bloom–bust lifecycle. A representative of this genus, Ruegeria sp. R11, is known to cause bleaching disease in a red macroalga at elevated temperatures. To investigate if the pathogenicity of R11 extends to microalgae, it was co-cultured with several cell types of E. huxleyi near the alga’s optimum (18°C), and at an elevated temperature (25°C) known to induce virulence in R11. The algal populations were monitored using flow cytometry and pulse-amplitude modulated fluorometry. Cultures of algae without bacteria remained healthy at 18°C, but lower cell counts in control cultures at 25°C indicated some stress at the elevated temperature. Both the C (coccolith-bearing) and S (scale-bearing swarming) cell types of E. huxleyi experienced a rapid decline resulting in apparent death when co-cultured with R11 at 25°C, but had no effect on N (naked) cell type at either temperature. R11 had no initial negative impact on C and S type E. huxleyi population size or health at 18°C, but caused death in older co-cultures. This differential effect of R11 on its host at 18 and 25°C suggest it is a temperature-enhanced opportunistic pathogen of E. huxleyi. We also detected caspase-like activity in dying C type cells co-cultured with R11, which suggests that programmed cell death plays a role in the death of E. huxleyi triggered by R11 – a mechanism induced by viruses (EhVs) and implicated in E. huxleyi bloom collapse. Given that E. huxleyi has

Analyses and comparison of a novel, hybrid, multifunctional orthopedic composite and implant

Science.gov (United States)

Dicicco, Michael

In the orthopedic/medical device industry, 2, 2' -bis-(4-(2-hydroxy-3-methacryloxypropoxy)phenyl]propane (Bis-GMA)- and diurethanedimethacrylate (DUDMA)-based polymeric biomaterials have become well-known substitutes for polymethylmethacrylate (PMMA)- and ultra-high molecular weight polyethylene (UHMWPE)-based biomaterials, respectively. The development of these polymeric biomaterials cannot continue without direct comparison studies against currently marketed materials. The initiative for this research stems wholly from developing analytical methodologies that assist in qualifying novel biomaterials under development, by evaluating their chemical properties, performance, and safety. The goals of this research were: (i) Characterize the assay/quality of every resin component and quantify elution of extractable monomers from novel, DUDMA-based RHAKOSS(TM) implant, (ii) Determine degree of conversion (alpha) and rate of polymerization (Rp) for novel, Bis-GMA-based CORTOSS(TM) composite, (iii) Assess risk for radical-induced post-surgical cytotoxicity for CORTOSS, (iv) Determine if surface radical chemistries occur for sterilized RHAKOSS and assess its oxidative stability, and (v) Quantify antibiotic elution from antibiotic-impregnated CORTOSS and identify factors that control elution. The phenomena studied necessitated the utilization of several analytical spectroscopic techniques; fluorometry, differential scanning calorimetry (DSC), electron paramagnetic resonance (EPR), ultraviolet-visible (UV-VIS), attenuated total reflectance---Fourier-transform infrared (ATR-FTIR), and high performance liquid chromatography (HPLC). A battery of lateral chemical techniques were employed as well; including molecular derivatization/tagging, phase partitioning, spin-trapping, and thermal annealing. Results demonstrated that RHAKOSS monomer percent compositions were prepared according to formulations and monomer elution was virtually undetectable, serving as an empirical gauge to

Quantitative transient GUS expression in J-104 rice calli through manipulation of in vitro culture conditions.

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Maylin Pérez Bernal

2009-10-01

This paper purposes suitable conditions for callus induction and co-cultivation with Agrobacterium tumefaciens of J-104 rice cultivar. It was evaluated the effect of different concentrations of 2.4-D and agar, and the inclusion of L-proline and L-glutamine in callus culture medium. The use of 2.5 mg/L 2.4-D and 0.8% agar allowed the highest percentage of embryogenic calli. Callus formation was improved considerably with 500 mg/L of L-proline and L-glutamine in the culture medium. Different factors were studied throughout co-cultivation of calli with A. tumefaciens: inoculation time, co-cultivation temperature, concentration of acetosyringone and co-cultivation period. Transient GUS expression was quantified by fluorometry in all co-cultivated calli. The best results were obtained with the following conditions: 10 min as inoculation time, 100µM acetosyringone in co-cultivation medium, temperature of 20ºC, and 3 days as co-cultivation period. Key words: Agar; callus; co-cultivation; fluorometric GUS activity. Resumen Se describen las condiciones óptimas para la callogénesis y cocultivo de callos con Agrobacterium tume-faciens de la variedad de arroz J-104. Se determinó el efecto de diferentes concentraciones de 2.4-D, agar y de L-prolina y L-glutamina en el medio de cultivo de callos. El uso de 2,5 mg/L de 2.4-D y 0,8% de agar permitió lograr el porcentaje más alto de callos embriogénicos. La formación de callos fue mejorada considerablemente con la adición de 500 mg/L de L-prolina e igual concentración de L-glutamina en el medio de cultivo. Se estudiaron diferentes factores en el cocultivo de los callos con A. tumefaciens: tiempo de inoculación, concentración de acetosiringona, temperatura y tiempo de cocultivo. Para comparar el efecto de cada factor sobre la expresión GUS se cuantificó la actividad transitoria mediante fluorimetría. Los valores más altos de actividad fluorimétrica fueron obtenidos con las siguientes condiciones: 10 min de

Traditional Chinese medicine Guizhi Fuling capsule used for therapy of dysmenorrhea via attenuating uterus contraction.

Science.gov (United States)

Sun, Lan; Liu, Lina; Zong, Shaobo; Wang, Zhengzhong; Zhou, Jun; Xu, Zhiliang; Ding, Gang; Xiao, Wei; Kou, Junping

2016-09-15

Guizhi Fuling formula, a well-known Chinese herbal formula recorded in the Eastern Han Dynasty, is composed of Cinnamomum cassia (L.) J.Presl (Cassia bark), Poria cocos (Schw.) Wolf (Poria), Paeonia suffruticosa andrews (Moutan Cortex), Paeonia lactiflora Pall (Herbaceous peony), and Amygdalus persica L.(Persicae Semen). It has clinical efficacy of activating blood circulation to dissipate blood stasis and is commonly used for the treatment of primary dysmenorrhea. However, its therapeutic mechanism has not been clearly elucidated. The aim of this study is to reveal molecular mechanisms of action using in vivo and in vitro experimental models. The ICR mouse uterine contraction was induced by oxytocin exposure following estradiol benzoate pretreatment. Mice were given GZFLC (0.54, 1.08g/kg) by gavage. The levels of NO, PGF2α and Ca(2+) in uterine tissue were determined according to instructions. Cyclooxygenase-2 (COX-2) and oxytocin receptor (OTR) proteins in uterine tissue were assessed by Western Blot. Mouse isolated uterus strips were mounted in tissue organ baths containing Locke's solution. The contractile responses were recorded with Power Lab recording system. The effect of GZFLC on spontaneous uterine contraction, and uterine contraction induced by oxytocin, PGF2α was observed. Myometrial cells were exposed to oxytocin (5U/L) to induce calcium release, and the effect of GZFLC and its components (PL, PGG, CA) on intracellular Ca(2+) was analyzed with fluorometry imaging. In vivo study demonstrated that GZFLC significantly reduced oxytocin-induced writhing responses with a maximal inhibition of 55%. It also decreased the levels of NO, PGF2α and Ca(2+) in oxytocin-induced mice uterine tissue. Moreover, Western blot analysis showed that COX-2 and OTR expressions in uterine tissue of dysmenorrhea mice were significantly reduced. GZFLC inhibited spontaneous uterus contractions in a dose-dependent manner, and the IC50 value was 0.99mg/ml. The IC50 values of

A comparative assessment of antiproliferative properties of resveratrol and ethanol leaf extract of Anogeissus leiocarpus (DC) Guill and Perr against HepG2 hepatocarcinoma cells.

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Olugbami, Jeremiah Olorunjuwon; Damoiseaux, Robert; France, Bryan; Onibiyo, Esther Modupe; Gbadegesin, Michael Adedapo; Sharma, Shivani; Gimzewski, James Kazimierz; Odunola, Oyeronke Adunni

2017-08-02

Epidemiological and experimental evidences have shown cancer as a leading cause of death worldwide. Although the folklore use of plants as a reliable source of health-restoring principles is well-documented, the search for more of such plants that are active against diseases, such as cancer, continues. We report here a laboratory-based evidence of the relevance of an ethanol leaf extract of Anogeissus leiocarpus (A2L) in comparison with resveratrol, a natural polyphenol, in cancer therapy. The quantitative assessment of flavonoid and phenolic contents involved quercetin and gallic acid as standards, respectively were determined using spectrophotometry. Cytotoxicity was determined fluorometrically using propidium-iodide-staining method. Antioxidant status, adenosine triphosphate (ATP) levels, caspase activities and mitochondrial integrity were assessed using fluorometry/luminometry. The antioxidant assay demonstrated that A2L possesses a strong antioxidant capacity as compared with the reference compounds, ascorbic acid and butylated hydroxytoluene. This is further buttressed by the significantly high level of phenolics obtained in the quantitative assessment of the extract. A 72-h post-treatment examination indicated that both A2L and resveratrol modulate the proliferation of HepG2 liver carcinoma cells in a time- and concentration-dependent manner. Determination of the total nuclei area, propidium-iodide negative and positive nuclei areas all further buttress the modulation of cell proliferation by A2L and resveratrol with the indication that the observed cell death is due to apoptosis and necrosis at lower and higher concentrations of treatments respectively. At lower concentrations (0.39-3.13 μg/mL), resveratrol possesses higher tendencies to activate caspases 3 and 7. Bioenergetically, both resveratrol and A2L do not adversely affect the cells at lower concentrations (0.39-6.25 μg/mL for resveratrol and 12.5-100.0 μg/mL for A2L) except at higher

Lethal and sub-lethal chronic effects of the herbicide diuron on seagrass

Energy Technology Data Exchange (ETDEWEB)

Negri, Andrew P., E-mail: a.negri@aims.gov.au [Australian Institute of Marine Science, Townsville, Queensland 4810 (Australia); Flores, Florita [Australian Institute of Marine Science, Townsville, Queensland 4810 (Australia); Mercurio, Phil [Australian Institute of Marine Science, Townsville, Queensland 4810 (Australia); University of Queensland and National Research Centre for Environmental Toxicology, Coopers Plains, Queensland 4108 (Australia); Mueller, Jochen F. [University of Queensland and National Research Centre for Environmental Toxicology, Coopers Plains, Queensland 4108 (Australia); Collier, Catherine J. [Centre for Tropical Water & Aquatic Ecosystem Research (TropWATER), James Cook University, Cairns, Queensland 4870 (Australia)

2015-08-15

Highlights: • We performed chronic exposures of two seagrass species to the herbicide diuron. • Diuron affected photosystem II (PSII) at 0.3 μg l{sup −1} and growth at 7.2 μg l{sup −1}. • Biomarkers indicated that carbon-assimilation from photosynthesis dropped following 0.6 μg l{sup −1} diuron exposure. • Energetic reserves in the seagrass were halved at 1.7 μg l{sup −1} after 11 weeks. • Chronic exposure to diuron is likely to enhance the impacts of low light stress during flood plumes - Abstract: Photosystem II herbicides from agricultural sources have been detected throughout nearshore tropical habitats including seagrass meadows. While PSII herbicides have been shown to inhibit growth in microalgae at low concentrations, the potential impacts of chronic low concentration exposures to seagrass health and growth have not been investigated. Here we exposed two tropical seagrass species Halodule uninervis and Zostera muelleri to elevated diuron concentrations (from 0.3 to 7.2 μg l{sup −1}) over a 79-day period followed by a 2-week recovery period in uncontaminated seawater. PAM fluorometry demonstrated rapid effect of diuron on photosystem II (PSII) in both seagrass species at 0.3 μg l{sup −1}. This effect included significant inhibition of photosynthetic efficiency (ΔF/F{sub m}′) and inactivation of PSII (F{sub v}/F{sub m}) over the 11 week exposure period. Significant mortality and reductions in growth was only observed at the highest exposure concentration of 7.2 μg l{sup −1} diuron. However, biochemical indicators demonstrated that the health of seagrass after this prolonged exposure was significantly compromised at lower concentrations. For example, the drop in C:N ratios (0.6 μg l{sup −1}) and reduced δ{sup 13}C (1.7 μg l{sup −1}) in seagrass leaves indicated reduced C-assimilation from photosynthesis. Critically, the energetic reserves of the plants (as measured by starch content in the root-rhizome complex) were

Lethal and sub-lethal chronic effects of the herbicide diuron on seagrass

International Nuclear Information System (INIS)

Negri, Andrew P.; Flores, Florita; Mercurio, Phil; Mueller, Jochen F.; Collier, Catherine J.

2015-01-01

Highlights: • We performed chronic exposures of two seagrass species to the herbicide diuron. • Diuron affected photosystem II (PSII) at 0.3 μg l −1 and growth at 7.2 μg l −1 . • Biomarkers indicated that carbon-assimilation from photosynthesis dropped following 0.6 μg l −1 diuron exposure. • Energetic reserves in the seagrass were halved at 1.7 μg l −1 after 11 weeks. • Chronic exposure to diuron is likely to enhance the impacts of low light stress during flood plumes - Abstract: Photosystem II herbicides from agricultural sources have been detected throughout nearshore tropical habitats including seagrass meadows. While PSII herbicides have been shown to inhibit growth in microalgae at low concentrations, the potential impacts of chronic low concentration exposures to seagrass health and growth have not been investigated. Here we exposed two tropical seagrass species Halodule uninervis and Zostera muelleri to elevated diuron concentrations (from 0.3 to 7.2 μg l −1 ) over a 79-day period followed by a 2-week recovery period in uncontaminated seawater. PAM fluorometry demonstrated rapid effect of diuron on photosystem II (PSII) in both seagrass species at 0.3 μg l −1 . This effect included significant inhibition of photosynthetic efficiency (ΔF/F m ′) and inactivation of PSII (F v /F m ) over the 11 week exposure period. Significant mortality and reductions in growth was only observed at the highest exposure concentration of 7.2 μg l −1 diuron. However, biochemical indicators demonstrated that the health of seagrass after this prolonged exposure was significantly compromised at lower concentrations. For example, the drop in C:N ratios (0.6 μg l −1 ) and reduced δ 13 C (1.7 μg l −1 ) in seagrass leaves indicated reduced C-assimilation from photosynthesis. Critically, the energetic reserves of the plants (as measured by starch content in the root-rhizome complex) were approximately halved following diuron exposure at and above

Development of novel encapsulated formulations using albumin-chitosan as a polymer matrix for ocular drug delivery

Science.gov (United States)

Addo, Richard Tettey

exposed up to 120 minutes to different BSA-CSN MS concentrations. Using fluorometry, the influence of temperature and effect of metabolic inhibition were studied. The in vitro uptake and internalization studies were evaluated using confocal microscopy in HCET-1. In vivo studies were evaluated in rabbit's eye using blink response and pupil to cornea ratio for tetracaine and atropine studies respectively. Results: Our results showed particles size in the range of 3-5 microns with encapsulation efficiency of about 96 percent. Differential Scanning Calorimetry showed no drug-polymer interactions. BSA-CSN MS were internalized by the HCET-1 and was affected both by temperature and metabolic inhibitor, sodium azide. There were no signs of ocular surface toxicity or inflammation. The encapsulated drugs exhibited superior properties in vivo compared to the solution formulations currently in clinical use. Conclusion: We successfully developed microparticulate drug carriers for ocular delivery. BSA-CSN MS were internalized by the HCET-1 by temperature dependent active transport mechanism that did not compromise cell viability.

Multiple sessions of liposomal doxorubicin delivery via focused ultrasound mediated blood-brain barrier disruption: a safety study.

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Aryal, Muna; Vykhodtseva, Natalia; Zhang, Yong-Zhi; McDannold, Nathan

2015-04-28

Transcranial MRI-guided focused ultrasound is a rapidly advancing method for delivering therapeutic and imaging agents to the brain. It has the ability to facilitate the passage of therapeutics from the vasculature to the brain parenchyma, which is normally protected by the blood-brain barrier (BBB). The method's main advantages are that it is both targeted and noninvasive, and that it can be easily repeated. Studies have shown that liposomal doxorubicin (Lipo-DOX), a chemotherapy agent with promise for tumors in the central nervous system, can be delivered into the brain across BBB. However, prior studies have suggested that doxorubicin can be significantly neurotoxic, even at small concentrations. Here, we studied whether multiple sessions of Lipo-DOX administered after FUS-induced BBB disruption (FUS-BBBD) induces severe adverse events in the normal brain tissues. First, we used fluorometry to measure the doxorubicin concentrations in the brain after FUS-BBBD to ensure that a clinically relevant doxorubicin concentration was achieved in the brain. Next, we performed three weekly sessions with FUS-BBBD±Lipo-DOX administration. Five to twelve targets were sonicated each week, following a schedule described previously in a survival study in glioma-bearing rats (Aryal et al., 2013). Five rats received three weekly sessions where i.v. injected Lipo-DOX was combined with FUS-BBBD; an additional four rats received FUS-BBBD only. Animals were euthanized 70days from the first session and brains were examined in histology. We found that clinically-relevant concentrations of doxorubicin (4.8±0.5μg/g) were delivered to the brain with the sonication parameters (0.69MHz; 0.55-0.81MPa; 10ms bursts; 1Hz PRF; 60s duration), microbubble concentration (Definity, 10μl/kg), and the administered Lipo-DOX dose (5.67mg/kg) used. The resulting concentration of Lipo-DOX was reduced by 32% when it was injected 10min after the last sonication compared to cases where the agent was

Living in a digital world: features and applications of FPGA in photon detection

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Arnesano, Cosimo

Optical spectroscopy and imaging outcomes rely upon many factors; one of the most critical is the photon acquisition and processing method employed. For some types of measurements it may be crucial to acquire every single photon quickly with temporal resolution, but in other cases it is important to acquire as many photons as possible, regardless of the time information about each of them. Fluorescence Lifetime Imaging Microscopy belongs to the first case, where the information of the time of arrival of every single photon in every single pixel is fundamental in obtaining the desired information. Spectral tissue imaging belongs to the second case, where high photon density is needed in order to calculate the optical parameters necessary to build the spectral image. In both cases, the current instrumentation suffers from limitations in terms of acquisition time, duty cycle, cost, and radio-frequency interference and emission. We developed the Digital Frequency-Domain approach for photon acquisition and processing purpose using new digital technology. This approach is based on the use of photon detectors in photon counting mode, and the digital heterodyning method to acquire data which is analyzed in the frequency domain to provide the information of the time of arrival of the photons . In conjunction with the use of pulsed laser sources, this method allows the determination of the time of arrival of the photons using the harmonic content of the frequency domain analysis. The parallel digital FD design is a powerful approach that others the possibility to implement a variety of different applications in fluorescence spectroscopy and microscopy. It can be applied to fluorometry, Fluorescence Lifetime Imaging (FLIM), and Fluorescence Correlation Spectroscopy (FCS), as well as multi frequency and multi wavelength tissue imaging in compact portable medical devices. It dramatically reduces the acquisition time from the several minutes scale to the seconds scale, performs

Assessment of in vivo fluorescence method for chlorophyll-a estimation in optically complex waters (Curuai floodplain, Pará - Brazil Avaliação do método de fluorescência in vivo para a estimativa da concentração de clorofila-a em águas opticamente complexas (planície de inundação do Curuai, Pará - Brasil

Directory of Open Access Journals (Sweden)

Rafael Damiati Ferreira

2012-12-01

Full Text Available AIM: This paper describes an experiment carried out to evaluate in vivo fluorescence (IVF as an alternative method for chlorophyll-a estimation in optically complex aquatic environment (Amazon floodplain lakes METHODS: The experiment consisted of collecting in situ measurements at 26 sampling stations distributed throughout Curuai floodplain lakes. For each sampling station the following parameters were measured: temperature, turbidity, depth, Secchi depth, chlorophyll-a (Chl-a concentration, total suspended solids (TSS and dissolved organic carbon (DOC, concurrently with several transects of IVF. Two methods were tested for quantifying the fluorescence measurement to be used as input for the chlorophyll-a estimates: instantaneous IFV and average IVF. Global and regional models were tested and assessed by analyzing optically active components (Chl-a, DOC and TSS of the water. RESULTS: Regardless of fluorescence estimating method, the results indicate that it was not possible to fit a global model for estimating Chl-a from IVF for all the lakes in the Curuai floodplain. Regional models provided contrasting results according to the concentration of optically active components. The best results were observed for aquatic systems with a single dominant component homogenously distributed throughout the lake. The results highlight the influence of the ratios Chl-a/TSS, Chl-a/DOC and Phaephytin/Chl-a in the relationship between IVF and chlorophyll concentration. CONCLUSIONS: It was not possible to develop a global model to account for the entire region of Curuai floodplain. The search for regional models provided insights on the main factors affecting the relationship between IVF and Chl-a concentration. Nevertheless this work reinforces the great potential of fluorometry technique, since even with a small number of samples it was possible to set a good model in the main lake of the Curuai floodplain. In spite the fact that this is not an accurate

Radioisotopes and Radiation in Animal and Plant Insect Pest Control; Emploi des radioisotopes et des rayonnements dans la lutte contre les insectes nuisibles aux plantes et aux animaux; Ispol'zovanie radioizotopov i radiashchi v bor'be s nasekomymi-vreditelyami rastenij i zhivotnykh; Utilizacion de los radioisotopos y de las radiaciones en la lucha contra los insectos nocivos para las plantas y los animales

Energy Technology Data Exchange (ETDEWEB)

Andreev, S. V.; Martens, B. K.; Samojlova, V. A.; Molchanova, Z. I. [Vsesoyuznyj Institut Zashchity Rastenij, Leningrad, SSSR (Russian Federation)

1963-09-15

agnatus Crow et Pseudogonia cinerascens Rond.) a permis d'etablir l'ampleur des migrations, de determiner remplacement des foyers de reproduction et l'importance numerique des populations et d'etudier le mode d'alimentation des parasites. Par la meme methode, on a etabli la dynamique de la propagation du doryphore (Leptinotarsa decemlineota Say), ce qui a permis d'obtenir les donnees indispensables pour proceder a leur extermination. En outre, cette methode permet d'etudier diverses questions extremement compliquees ayant trait au parasitisme et au pillage (etude des cycles alimentaires de l'alimentation interimaire et de la chimiotaxie.). Grace a l'emploi de la methode d'automarquage des rongeurs des champs a l'aide d'appats radioactifs, on a pu determiner les dimensions des zones ou ils se procurent leur nourriture. L'utilisation de parasiticides marques conjointement avec les methodes de radiochromatographie, de fluorometrie, etc. - a permis d'etudier les processus biochimiques les plus complexes qui interviennent dans les plantes et chez les insectes soie l'effet des substances toxiques. Se fondant sur l'exemple des composes organiques du phosphore du type ''tiophosphate'' et ''metaphosphate'', on a determine la vitesse de l'hydrolyse de ces insecticides en fonction de la phase de developpement de la plante, de son etat physiologique et des conditions du milieu ambiant. On a egalement determine la duree de remanence des residus de substances toxiques sur les produits agricoles, en fonction de celle des traitements chimiques. Les radioisotopes ont aussi revele les particularites des processus metaboliques qui s'effectuent chez les plantes qui resistent et celles qui ne resistent pas aux herbicides. Le marquage des parasiticides a l'aide de radioisotopes de courte periode permet d'evaluer l'efficacite du traitement des graines et semences dans differentes conditions de pulverisation, et quels parasiticides conviennent le mieux pour un traitement donne. L'action des