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Sample records for fluorimetry

  1. Differential scanning fluorimetry illuminates silk feedstock stability and processability

    Czech Academy of Sciences Publication Activity Database

    Dicko, C.; Kasoju, Naresh; Hawkins, N.; Vollrath, F.

    2016-01-01

    Roč. 12, č. 1 (2016), s. 255-262 ISSN 1744-683X R&D Projects: GA MŠk(CZ) EE2.3.30.0029 Institutional support: RVO:61389013 Keywords : differential scanning fluorimetry * biomaterials * silk fibroin Subject RIV: CE - Biochemistry Impact factor: 3.889, year: 2016

  2. Laser fluorimetry of mixtures of polyatomic organic compounds using artificial neural networks

    International Nuclear Information System (INIS)

    Dolenko, S A; Gerdova, I V; Dolenko, T A; Fadeev, V V

    2001-01-01

    New possibilities of laser fluorimetry offered by the use of algorithms for solving inverse problems based on artificial neural networks are demonstrated. A two-component mixture of polyatomic organic compounds is analysed by three methods of laser fluorimetry: a direct analysis of the fluorescence band, the kinetic fluorimetry (when durations of the laser pulse and the detector gate pulse are comparable with the fluorescence lifetimes or exceed them), and the saturation fluorimetry. The numerical experiments showed that the use of artificial neural networks in these methods provides a high practical stability of the solution of inverse problems and ensures a high sensitivity and a high accuracy of determining the contribution of components to fluorescence and of measuring molecular photophysical parameters, which can be used for the identification of components. (laser applications and other topics in quantum electronics)

  3. Fluorimetry as a Simple and Sensitive Method for Determination of Catalase

    Directory of Open Access Journals (Sweden)

    Mehdi Hedayati

    2014-02-01

    Full Text Available Background: Catalase enzyme plays an important role in the anti-oxidation defense of body so it is important to measure its activity. Nowadays catalase activity measurement is performed by expensive imported kits in various scientific fields. The purpose of this study was to design a sensitive fluorimetry method for measuring catalase activity with improved sensitivity, accuracy and speed. Materials and Methods: In this study, the reaction of hydrogen peroxide with peroxidase (as a reaction accelerator was used in fluorimetry for catalase activity measuring in serum samples in order to increase the sensitivity of the assay. The sensitivity and intra- and inter-assay accuracy, verification test, recovery and parallelism tests, comparison method and correlation and coherence investigation methods were also performed. In order to increase the accuracy and speed of reading, the assay was performed in microplates and reading was done in fluorimetry plates. Results: The percentage of intra- and inter-assay variation coefficients were measured 3.8- 6.6 % and 4.1-7.3%, respectively. Comparison of the results of mentioned method for 50 serum samples with common colorimetric method showed a good correlation (0.917. In assessing the accuracy, the recovery percent was obtained 91% to 107%. The test sensitivity was measured 0.02 IU/ml. Conclusion: The fluorimetry method by microplate reading has a sufficient precision, accuracy and efficiency for catalase activity measuring as well as speed of measurement. Thus it can be an alternative method to conventional imported colorimetric methods.

  4. The fluorimetry for control of internal contamination of exposed workers to natural and enriched uranium

    International Nuclear Information System (INIS)

    Gaburo, J.C.; Todo, A.S.; Sordi, G.M.A.A.

    2000-01-01

    This study is a part of bioassay program revision applied to the uranium processing plants at IPEN-CNEN/SP. The workers of these facilities handle both natural uranium and uranium compounds with different isotopic composition which could reach up to 20% in 235 U. The most commonly employed techniques for the determination of uranium in urine at IPEN are fluorimetry and alpha spectrometry with detection limit of 1.0 mgL-1. and 1,0 mBqL-1 , respectively. Based in advantages and disadvantages of each technique it is very important to identify the workers groups that should be submitted for these analysis. In this report a limiting value of uranium concentration in urine, mgL-1, obtained by fluorimetry is proposed. All the results greater than these limiting value indicate the necessity to carry out a additional measurement by alpha spectroscopy. The uranium mass that result in a pre-determined limit committed effective dose is function of isotopic composition. Consequently, the predicted value of the measured of urinary excretion is function of isotopic composition also and depends of absorption characteristics when inhaled and of the monitoring interval considered. In this report the uranium concentration values for reference levels and limits doses are determined. Based on these results the procedures to use the fluorimetry or both fluorimetry and alpha-spectrometry were adopted. (author)

  5. Low cost, microcontroller based heating device for multi-wavelength differential scanning fluorimetry.

    Science.gov (United States)

    Hoeser, Jo; Gnandt, Emmanuel; Friedrich, Thorsten

    2018-01-23

    Differential scanning fluorimetry is a popular method to estimate the stability of a protein in distinct buffer conditions by determining its 'melting point'. The method requires a temperature controlled fluorescence spectrometer or a RT-PCR machine. Here, we introduce a low-budget version of a microcontroller based heating device implemented into a 96-well plate reader that is connected to a standard fluorescence spectrometer. We demonstrate its potential to determine the 'melting point' of soluble and membranous proteins at various buffer conditions.

  6. Estimation of uranium in bioassay samples of occupational workers by laser fluorimetry

    International Nuclear Information System (INIS)

    Suja, A.; Prabhu, S.P.; Sawant, P.D.; Sarkar, P.K.; Tiwari, A.K.; Sharma, R.

    2012-01-01

    A newly established uranium processing facility has been commissioned at BARC, Trombay. Monitoring of occupational workers is essential to assess intake of uranium in this facility. A group of 21 workers was selected for bioassay monitoring to assess the existing urinary excretion levels of uranium before the commencement of actual work. Bioassay samples collected from these workers were analyzed by ion-exchange technique followed by laser fluorimetry. Standard addition method was followed for estimation of uranium concentration in the samples. The minimum detectable activity by this technique is about 0.2 ng. The range of uranium observed in these samples varies from 19 to 132 ng/L. Few of these samples were also analyzed by fission track analysis technique and the results were found to be comparable to those obtained by laser fluorimetry. The urinary excretion rate observed for the individual can be regarded as a 'personal baseline' and will be treated as the existing level of uranium in urine for these workers at the facility. (author)

  7. Liquid-solid extraction of uranium (VI) with TOPO - molten naphthalene and determination by laser fluorimetry in geological samples

    International Nuclear Information System (INIS)

    Kumar, Sanjay; Krishnakumar, M.; Patwardhan, A.A.

    2007-01-01

    A simple, rapid, sensitive, cost-effective and efficient method for separation of uranium using tri-n-octylphosphine oxide (TOPO)-molten naphthalene as solid phase extractant and its determination by laser fluorimetry in geological samples (rock, soil, sediment) was developed. Under optimum conditions, using 50 mg TOPO and 100 mg naphthalene, 50 - 5000 ng of uranium in 10 ml sample solution (3% (v/v) HNO 3 ) could be extracted quantitatively. The extracted uranium was stripped using tetra sodium pyrophosphate (5% (v/v) solution, pH adjusted to 7.0 with H 3 PO 4 ) and determined by laser fluorimetry. The influence of different acid concentrations, the amount of solid phase extractant, sample volumes, different stripping reagents, their volumes and effect of foreign ions on the extraction and determination of uranium (VI) were investigated. Synthetic samples of varying concentration as regards uranium were prepared and analysed. Recoveries ranging from 90% to 105% were obtained. The method was validated by analyzing four certified reference materials namely, BL-5, DH-1a, SY-2, SY-3 and the values obtained for uranium agreed well with the certified values. The method was also applied to the determination of uranium in geological samples (rock, soil and sediment) by laser fluorimetry and the results obtained compared favorably with those obtained from the pellet fluorimetry method. Following the proposed method, determination limit for uranium was found to be 1 μg/g with RSD ± 10%. (author)

  8. LED vs laser fluorimetry: a comparative study for the determination of uranium in natural waters

    International Nuclear Information System (INIS)

    Shenoy, N.; Parab, H.; Sounderajan, S.; Kiran Kumar; Kumar, S.D.; Reddy, A.V.R.

    2015-01-01

    Measurement of uranium in water samples has acquired considerable importance ever since its occurrence in drinking water sources was reported. Among the various methods available for uranium quantification at ultra trace levels, laser fluorimetry (LF) method is the method of choice due to its simplicity, speed and high sensitivity compared to other analytical techniques. This technique is based on the measurement of fluorescence of uranium complexes in aqueous solution. Recently, laser source has been replaced by light emitting diode (LEDs) in the fluorimeter systems. In comparison to laser source, LED source is, cost effective, generates less heat and has extended lifetime. Herein, authors have presented a comparison of LED based fluorimeter (Quantalase, Indore, India) and laser fluorimeter (CAT, Indore, India) for the determination of uranium in natural waters

  9. Th, U and trace elements determination in Egyiptian Lake sediments by INAA and laser fluorimetry

    International Nuclear Information System (INIS)

    Ismail, S.S.; Grass, F.; Ghods, A.

    1995-01-01

    A study was undertaken to determine element concentrations in Aswan High Dam Lake sediments. Sediment samples were collected from 40 to 500 km upstream of the dam to follow the sedimentation process and the distribution of Th, U and the trace elements in the lake. INAA was applied for the determination of Sm, Ce, Lu, Th, Cr, Yb, Au, Hf, Ba, Nd, Cs, Tb, Sc, Rb, Fe, Zn, Co, Eu, and Sb, while Laser Fluorimetry was applied for U determination. The accuracy and the reproducibility of the techniques were tested with IAEA standard materials (SL, Soil-7). The U values ranged from 4 ppm to 18 ppm, Th values were between 2 and 10 ppm, and showed a very good correlation with the rare earth elements and Fe. The distribution of most of the elements in the lake follows the same trend as the distribution of the clays in the sediments. Ba showed a negative correlation with most of the elements under investigation. (author) 5 refs.; 4 figs.; 7 tabs

  10. Study on inclusion complex of cyclodextrin with methyl xanthine derivatives by fluorimetry

    Science.gov (United States)

    Wei, Yan-Li; Ding, Li-Hua; Dong, Chuan; Niu, Wei-Ping; Shuang, Shao-Min

    2003-10-01

    The inclusion complexes of β-cyclodextrin (β-CD) and HP-β-cyclodextrin (HP-β-CD) with caffeine, theophylline and theobromine were investigated by fluorimetry. Various factors affecting the formation of inclusion complexes were discussed in detail including forming time, pH effect and temperature. The results indicate that inclusion process was affected seriously by laying time and pH. The forming time of β-CD inclusion complexes is much longer than that of HP-β-CD. The optimum pH range is about 7-12 for caffeine, 8-10 for TP, 10.5-12 for TB. The intensities of their fluorescence increase with the decreasing of temperature. Their maximum excitation wavelengths are all in the range of 280-290 nm. The emission wavelength of caffeine and theophylline are both in the range of 340-360 nm, and that of theobromine is about 325 nm. The fluorescence signals are intensified with the increasing concentration of CD. The stoichiometry of the inclusion complexes of CD with these three methyl xanthine derivatives are all 1:1 and the formation constant are all calculated.

  11. Comparison between fluorimetry and oximetry techniques to measure photosynthesis in the diatom Skeletonema costatum cultivated under simulated seasonal conditions.

    Science.gov (United States)

    Lefebvre, Sébastien; Mouget, Jean-Luc; Loret, Pascale; Rosa, Philippe; Tremblin, Gérard

    2007-02-01

    This study reports comparison of two techniques measuring photosynthesis in the ubiquitous diatom Skeletonema costatum, i.e., the classical oximetry and the recent modulated fluorimetry. Microalgae in semi-continuous cultures were exposed to five different environmental conditions simulating a seasonal effect with co-varying temperature, photoperiod and incident light. Photosynthesis was assessed by gross rate of oxygen evolution (P(B)) and the electron transport rate (ETR) measurements. The two techniques were linearly related within seasonal treatments along the course of the P/E curves. The light saturation intensity parameters (Ek and Ek(ETR)), and the maximum electron transport rate increased significantly with the progression of the season while the maximum light utilization efficiency for ETR (alpha(ETR)) was constant. By contrast, the maximum gross oxygen photosynthetic capacity (Pmax(B)) and the maximum light utilization efficiency for P(B) (alpha(B)) increased from December to May treatment but decreased from May to July treatment. Both techniques showed clear photoacclimation in microalgae with the progression of the season, as illustrated by changes in photosynthetic parameters. The relationship between the two techniques changed when high temperature, photoperiod and incident light were combined, possibly due to an overestimation of the PAR--averaged chlorophyll-specific absorption cross-section. Despite this change, our results illustrate the strong suitability of in vivo chlorophyll fluorimetry to estimate primary production in the field.

  12. The effects of LMWOAs on biodegradation of multi-component PAHs in aqueous solution using dual-wavelength fluorimetry

    International Nuclear Information System (INIS)

    Wei Xingyuan; Sang Lingzi; Chen Jianing; Zhu Yaxian; Zhang Yong

    2009-01-01

    Biodegradation of dissolved fluorene (Flu), phenanthrene (Ph) and pyrene (Py), three polycyclic aromatic hydrocarbons (PAHs), singly or as a mixture of the three, by two bacterial strains, MEBIC 5140 (Mycobacterium flavescens) and MEBIC 5141 (Mycobacterium scrofulaceum), as well as the effects of low molecular weight organic acids (LMWOAs), e.g. malic acid, citric acid and butyric acid on biodegradation of the three PAHs in mineral salts medium aqueous solution were investigated using a newly established dual-wavelength fluorimetric method. The results showed that biodegradation processes can be monitored simultaneously, quickly and simply by dual-wavelength fluorimetry. Both co-metabolism and inhibitory effects were found during the biodegradation of the three PAHs by MEBIC 5140 and MEBIC 5141. Positive effects of butyric acid and negative effects of citric acid on biodegradation of the three PAHs in a mixture were observed. - Biodegradation processes of dissolved multi-component PAHs in a mixture and effects of LMWOAs were investigated using a dual-wavelength fluorimetry.

  13. Determination of uranium in environmental samples from the Nile delta and the adjacent regions of Egypt using laser fluorimetry

    Energy Technology Data Exchange (ETDEWEB)

    Shawky, S; Ibrahim, N; Farouk, A [National research Center for Nuclear safety and radiation Control, Atomic Energy Authority, cairo (Egypt); Ghods, A [IAEA Laboratories, Chemistry Unit., A 244 Seibersdorf, (Austria)

    1995-10-01

    Total uranium content was determined in soil and plant samples taken from various areas in the Nile delta. Samples from east and west of the delta, the suez canal` cities and from alexandria region were analyzed using laser fluorimetry (LF). Uranium was extracted from digested ashed samples with methyl-isobutyl ketone (MIBK) and measured using laser fluorimeter. The radium content of the same soil samples was determined using gamma spectrometry (GS). The uranium content of plant samples was determined using (LF), since it has a detection limit lower than of (GS). The uranium content varied between 0.6-4.4{mu}g/g and 0.032-0.17 {mu}g/g for soil and plant samples respectively. 1 fig., 3 tabs.

  14. Determination of uranium in environmental samples from the Nile delta and the adjacent regions of Egypt using laser fluorimetry

    International Nuclear Information System (INIS)

    Shawky, S.; Ibrahim, N.; Farouk, A.; Ghods, A.

    1995-01-01

    Total uranium content was determined in soil and plant samples taken from various areas in the Nile delta. Samples from east and west of the delta, the suez canal' cities and from alexandria region were analyzed using laser fluorimetry (LF). Uranium was extracted from digested ashed samples with methyl-isobutyl ketone (MIBK) and measured using laser fluorimeter. The radium content of the same soil samples was determined using gamma spectrometry (GS). The uranium content of plant samples was determined using (LF), since it has a detection limit lower than of (GS). The uranium content varied between 0.6-4.4μg/g and 0.032-0.17 μg/g for soil and plant samples respectively. 1 fig., 3 tabs

  15. On-line determination of iodine in nuclear fuel reprocessing off-gas streams by a combination of laser-induced fluorimetry and laser photoacoustic spectroscopy

    International Nuclear Information System (INIS)

    Kuno, Yusuke; Sato, Souichi; Masui, Jinichi

    1992-01-01

    The on-line determination of molecular iodine and organic iodides in nuclear fuel reprocessing off-gas streams containing high concentrations of NO x gas was studied. Ultraviolet radiation is used to convert organic iodides into molecular iodine. The approximate concentration of iodine before and after the photochemical conversion in the presence of NO x gas was first determined by laser-induced fluorimetry. NO 2 was determined by photoacoustic spectroscopy, correcting the acoustic signal due to iodine by using the approximate iodine concentration. NO was determined from the concentrations of NO 2 before and after the photoirradiation based on the photochemical fraction changes of NO and NO 2 . The quenching of the fluorimetry due to NO and NO 2 was finally corrected with the NO and NO 2 concentrations obtained. The detection limit of the proposed method is 10 nl 1 -1 . 7 figs., 2 tabs., 11 refs

  16. Multi frequency phase fluorimetry (MFPF) for oxygen partial pressure measurement: ex vivo validation by polarographic clark-type electrode.

    Science.gov (United States)

    Boehme, Stefan; Duenges, Bastian; Klein, Klaus U; Hartwich, Volker; Mayr, Beate; Consiglio, Jolanda; Baumgardner, James E; Markstaller, Klaus; Basciani, Reto; Vogt, Andreas

    2013-01-01

    Measurement of partial pressure of oxygen (PO2) at high temporal resolution remains a technological challenge. This study introduces a novel PO2 sensing technology based on Multi-Frequency Phase Fluorimetry (MFPF). The aim was to validate MFPF against polarographic Clark-type electrode (CTE) PO2 measurements. MFPF technology was first investigated in N = 8 anaesthetised pigs at FIO2 of 0.21, 0.4, 0.6, 0.8 and 1.0. At each FIO2 level, blood samples were withdrawn and PO2 was measured in vitro with MFPF using two FOXY-AL300 probes immediately followed by CTE measurement. Secondly, MFPF-PO2 readings were compared to CTE in an artificial circulatory setup (human packed red blood cells, haematocrit of 30%). The impacts of temperature (20, 30, 40°C) and blood flow (0.8, 1.6, 2.4, 3.2, 4.0 L min(-1)) on MFPF-PO2 measurements were assessed. MFPF response time in the gas- and blood-phase was determined. Porcine MFPF-PO2 ranged from 63 to 749 mmHg; the corresponding CTE samples from 43 to 712 mmHg. Linear regression: CTE = 15.59+1.18*MFPF (R(2) = 0.93; P0.05). MFPF response-time (monoexponential) was 1.48±0.26 s for the gas-phase and 1.51±0.20 s for the blood-phase. MFPF-derived PO2 readings were reproducible and showed excellent correlation and good agreement with Clark-type electrode-based PO2 measurements. There was no relevant impact of temperature and blood flow upon MFPF-PO2 measurements. The response time of the MFPF FOXY-AL300 probe was adequate for real-time sensing in the blood phase.

  17. Multi Frequency Phase Fluorimetry (MFPF) for Oxygen Partial Pressure Measurement: Ex Vivo Validation by Polarographic Clark-Type Electrode

    Science.gov (United States)

    Boehme, Stefan; Duenges, Bastian; Klein, Klaus U.; Hartwich, Volker; Mayr, Beate; Consiglio, Jolanda; Baumgardner, James E.; Markstaller, Klaus; Basciani, Reto; Vogt, Andreas

    2013-01-01

    Background Measurement of partial pressure of oxygen (PO2) at high temporal resolution remains a technological challenge. This study introduces a novel PO2 sensing technology based on Multi-Frequency Phase Fluorimetry (MFPF). The aim was to validate MFPF against polarographic Clark-type electrode (CTE) PO2 measurements. Methodology/Principal Findings MFPF technology was first investigated in N = 8 anaesthetised pigs at FIO2 of 0.21, 0.4, 0.6, 0.8 and 1.0. At each FIO2 level, blood samples were withdrawn and PO2 was measured in vitro with MFPF using two FOXY-AL300 probes immediately followed by CTE measurement. Secondly, MFPF-PO2 readings were compared to CTE in an artificial circulatory setup (human packed red blood cells, haematocrit of 30%). The impacts of temperature (20, 30, 40°C) and blood flow (0.8, 1.6, 2.4, 3.2, 4.0 L min−1) on MFPF-PO2 measurements were assessed. MFPF response time in the gas- and blood-phase was determined. Porcine MFPF-PO2 ranged from 63 to 749 mmHg; the corresponding CTE samples from 43 to 712 mmHg. Linear regression: CTE = 15.59+1.18*MFPF (R2 = 0.93; PPO2 ranged from 20 to 567 mmHg and CTE samples from 11 to 575 mmHg. Linear regression: CTE = −8.73+1.05*MFPF (R2 = 0.99; PPO2 due to variations of temperature were less than 6 mmHg (range 0–140 mmHg) and less than 35 mmHg (range 140–750 mmHg); differences due to variations in blood flow were less than 15 mmHg (all P-values>0.05). MFPF response-time (monoexponential) was 1.48±0.26 s for the gas-phase and 1.51±0.20 s for the blood-phase. Conclusions/Significance MFPF-derived PO2 readings were reproducible and showed excellent correlation and good agreement with Clark-type electrode-based PO2 measurements. There was no relevant impact of temperature and blood flow upon MFPF-PO2 measurements. The response time of the MFPF FOXY-AL300 probe was adequate for real-time sensing in the blood phase. PMID:23565259

  18. LASER BIOLOGY AND MEDICINE: Application of laser fluorimetry for determining the influence of a single amino-acid substitution on the individual photophysical parameters of a fluorescent form of a fluorescent protein mRFP1

    Science.gov (United States)

    Banishev, A. A.; Vrzheshch, E. P.; Shirshin, E. A.

    2009-03-01

    Individual photophysical parameters of the chromophore of a fluorescent protein mRFP1 and its two mutants (amino-acid substitution at position 66 - mRFP1/ Q66C and mRFP1/Q66S proteins) are determined. For this purpose, apart from conventional methods of fluorimetry and spectrophotometry, nonlinear laser fluorimetry is used. It is shown that the individual extinction coefficients of the chromophore of proteins correlate (correlation coefficient above 0.9) with the volume of the substituted amino-acid residue at position 66 (similar to the positions of the absorption, fluorescence excitation and emission maxima).

  19. Comparison of laser fluorimetry, high resolution gamma-ray spectrometry and neutron activation analysis techniques for determination of uranium content in soil samples

    International Nuclear Information System (INIS)

    Ghods, A.; Asgharizadeh, F.; Salimi, B.; Abbasi, A.

    2004-01-01

    Much more concern is given nowadays for exposure of the world population to natural radiation especially to uranium since 57% of that exposure is due to radon-222, which is a member of uranium decay series. Most of the methods used for uranium determination is low concentration require either tedious separation and preconcentration or the accessibility to special instrumentation for detection of uranium at this low level. this study compares three techniques and methods for uranium analysis among different soil sample with variable uranium contents. Two of these techniques, neutron activation analysis and high resolution gamma-ray spectrometry , are non-destructive while the other, laser fluorimetry is done via chemical extraction of uranium. Analysis of standard materials is done also to control the quality and accuracy of the work. In spite of having quite variable ranges of detection limit, results obtained by high resolution gamma-ray spectrometry based on the assumption of having secular equilibrium between uranium and its daughters, which causes deviation whenever this condition was missed. For samples with reasonable uranium content, neutron activation analysis would be a rapid and reliable technique, while for low uranium content laser fluorimetry would be the most appropriate and accurate technique

  20. The fluorimetry for control of internal contamination of exposed workers to natural and enriched uranium; A fluorimetria para o controle da contaminacao interna dos trabalhadores expostos a uranio natural e enriquecido

    Energy Technology Data Exchange (ETDEWEB)

    Gaburo, J.C.; Todo, A.S.; Sordi, G.M.A.A. [Instituto de Pesquisas Energeticas e Nucleares (IPEN), Sao Paulo, SP (Brazil)]. E-mail: janetegc@net.ipen.br

    2000-07-01

    This study is a part of bioassay program revision applied to the uranium processing plants at IPEN-CNEN/SP. The workers of these facilities handle both natural uranium and uranium compounds with different isotopic composition which could reach up to 20% in 235 U. The most commonly employed techniques for the determination of uranium in urine at IPEN are fluorimetry and alpha spectrometry with detection limit of 1.0 mgL-1. and 1,0 mBqL-1 , respectively. Based in advantages and disadvantages of each technique it is very important to identify the workers groups that should be submitted for these analysis. In this report a limiting value of uranium concentration in urine, mgL-1, obtained by fluorimetry is proposed. All the results greater than these limiting value indicate the necessity to carry out a additional measurement by alpha spectroscopy. The uranium mass that result in a pre-determined limit committed effective dose is function of isotopic composition. Consequently, the predicted value of the measured of urinary excretion is function of isotopic composition also and depends of absorption characteristics when inhaled and of the monitoring interval considered. In this report the uranium concentration values for reference levels and limits doses are determined. Based on these results the procedures to use the fluorimetry or both fluorimetry and alpha-spectrometry were adopted. (author)

  1. Dissecting the salt dependence of the Tus-Ter protein-DNA complexes by high-throughput differential scanning fluorimetry of a GFP-tagged Tus.

    Science.gov (United States)

    Moreau, Morgane J J; Schaeffer, Patrick M

    2013-12-01

    The analysis of the salt dependence of protein-DNA complexes provides useful information about the non-specific electrostatic and sequence-specific parameters driving complex formation and stability. The differential scanning fluorimetry of GFP-tagged protein (DSF-GTP) assay has been geared with an automatic Tm peak recognition system and was applied for the high-throughput (HT) determination of salt-induced effects on the GFP-tagged DNA replication protein Tus in complex with various Ter and Ter-lock sequences. The system was designed to generate two-dimensional heat map profiles of Tus-GFP protein stability allowing for a comparative study of the effect of eight increasing salt concentrations on ten different Ter DNA species at once. The data obtained with the new HT DSF-GTP allowed precise dissection of the non-specific electrostatic and sequence-specific parameters driving Tus-Ter and Tus-Ter-lock complex formation and stability. The major factor increasing the thermal resistance of Tus-Ter-lock complexes in high-salt is the formation of the TT-lock, e.g. a 10-fold higher Kspe was obtained for Tus-GFP:Ter-lockB than for Tus-GFP:TerB. It is anticipated that the system can be easily adapted for the study of other protein-DNA complexes.

  2. Comparative study of Uranium estimation in drinking water samples of seismically active regions of NW-Himalayas, Himachal Pradesh and SW-Punjab, India using Laser Fluorimetry

    Energy Technology Data Exchange (ETDEWEB)

    Bajwa, B.; Arora, V.; Saini, K. [Guru Nanak Dev University, Amritsar (India)

    2014-07-01

    The Laser Fluorimetry Technique has been used for the microanalysis of uranium content in drinking water samples collected from different sources like the hand pumps and natural springs of seismically active regions of Chamba and Dharamshala, Himachal Pradesh, NW-Himalayas and Bathinda and Mansa districts of SW-Punjab, state, India. The purpose of this study was to investigate the uranium concentration levels of ground water being used for drinking purposes and to determine its health effects, if any, to the local population of these regions. In the present study 47 samples of drinking water collected from different villages of the seismic active belt of Chamba and Dharamshala region of Himachal Pradesh, India have been analyzed for chemical and radiological toxicity. Uranium concentration in drinking water sample of study region ranged between 2.7 μgL{sup -1} - 53.9 μgL{sup -1} with an average value of 20.1 μgL{sup -1}. In SW-Punjab, Uranium concentration in 76 drinking water samples has been found to vary between 0.13 μgL{sup -1} and 676 μgL{sup -1} with an average of 90.2 μgL{sup -1}. Data analysis reveals that, 19% samples of NW-Himalayas water have uranium concentration higher than recommended limit of 30 μgL{sup -1} (WHO, 2011) while none of the samples exceeds the threshold of 60 μgL{sup -1} recommended by AERB, DAE, India, 2004. On the other hand, 64% water samples of SW-Punjab have uranium concentration higher than recommended limit of 30 μgL{sup -1} (WHO, 2011) while 39% water samples exceeds the threshold of 60 μgL{sup -1} recommended by AERB, DAE, India, 2004. Document available in abstract form only. (authors)

  3. Computer processing of microscopic images of bacteria : morphometry and fluorimetry

    NARCIS (Netherlands)

    Wilkinson, Michael H.F.; Jansen, Gijsbert J.; Waaij, Dirk van der

    1994-01-01

    Several techniques that use computer analysis of microscopic images have been developed to study the complicated microbial flora in the human intestine, including measuring the shape and fluorescence intensity of bacteria. These techniques allow rapid assessment of changes in the intestinal flora

  4. X-ray fluorimetry in tests of the aerosol substances

    International Nuclear Information System (INIS)

    Walentek, A.; Cudny, W.; Marzec, J.; Pawlowski, Z.; Zaremba, K.

    1992-01-01

    Test results of aerosol samples from atmospheric air collected from membrane drains were presented. The ANFLU-2 analyzer constructed at the Warsaw Technical University were used for testing. (author). 3 figs, 1 tab

  5. Determination of bisphenol A in canned fatty foods by coacervative microextraction, liquid chromatography and fluorimetry.

    Science.gov (United States)

    Bendito, María Dolores Pérez; Bravo, Soledad Rubio; Reyes, María Loreto Lunar; Prieto, Amalia García

    2009-02-01

    Decanoic acid reverse micelle-based coacervates were used to provide simple, rapid and almost solventless extraction of bisphenol A (BPA) from canned fatty foods. The procedure involved the extraction of 200-400 mg of homogenised food sample with an aqueous solution containing 20% THF and 200 mg of decanoic acid, conditions under which the coacervate (around 550 microl) formed in situ and instantaneously. The overall sample treatment took about 30 min and several samples could be simultaneously treated using conventional laboratory equipment. No clean-up or solvent evaporation were required before determination of BPA by liquid chromatography and fluorescence detection. Recoveries in samples were between 90 and 99%, with relative standard deviations in the range 2-7%. The limit of quantification ranged 29-15 ng g(-1) for 200-400 mg of sample, being far below the current specific migration limit (SML) set by the European Commission (600 ng g(-1)). The method was successfully applied to the determination of BPA in the solid content of canned fish (from 20 to 129 ng g(-1)) and meat (from undetected to 37 ng g(-1)).

  6. Optimization of experimental conditions in uranium trace determination using laser time-resolved fluorimetry

    International Nuclear Information System (INIS)

    Baly, L.; Garcia, M.A.

    1996-01-01

    At the present paper a new sample excitation geometry is presented for the uranium trace determination in aqueous solutions by the Time-Resolved Laser-Induced Fluorescence. This new design introduces the laser radiation through the top side of the cell allowing the use of cells with two quartz sides, less expensive than commonly used at this experimental set. Optimization of the excitation conditions, temporal discrimination and spectral selection are presented

  7. Estimation of uranium in bioassay samples of occupational workers by laser fluorimetry

    International Nuclear Information System (INIS)

    Suja, A.; Prabhu, S.P.; Sawant, P.D.; Sarkar, P.K.; Tiwari, A.K.; Sharma, R.

    2010-01-01

    A newly established uranium processing facility has been commissioned at BARC, Trombay. Monitoring of occupational workers at regulars intervals is essential to assess intake of uranium by the workers in this facility. The design and engineering safety features of the plant are such that there is very low probability of uranium getting air borne during normal operations. However, the leakages from the system during routine maintenance of the plant may result in intake of uranium by workers. As per the new biokinetic model for uranium, 63% of uranium entering the blood stream gets directly excreted in urine. Therefore, bioassay monitoring (urinalysis) was recommended for these workers. A group of 21 workers was selected for bioassay monitoring to assess the existing urinary excretion levels of uranium before the commencement of actual work. For this purpose, sample collection kit along with an instruction slip was provided to the workers. Bioassay samples received were wet ashed with conc. nitric acid and hydrogen peroxide to break down the metabolized complexes of uranium and it was co-precipitated with calcium phosphate. Separation of uranium from the matrix was done using ion exchange technique and final activity quantification in these samples was done using laser fluorimeter (Quantalase, Model No. NFL/02). Calibration of the laser fluorimeter is done using 10 ppb uranium standard (WHO, France Ref. No. 180000). Verification of the system performance is done by measuring concentration of uranium in the standards (1 ppb to 100 ppb). Standard addition method was followed for estimation of uranium concentration in the samples. Uranyl ions present in the sample get excited by pulsed nitrogen laser at 337.1 nm, and on de-excitation emit fluorescence light (540 nm) intensity which is measured by the PMT. To estimate the uranium in the bioassay samples, a known aliquot of the sample was mixed with 5% sodium pyrophosphate and fluorescence intensity was measured

  8. Determination of uranium in urine samples for workers in the phosphoric acid purification using fluorimetry technique

    International Nuclear Information System (INIS)

    Kharita, M. H.; Sakhita, Kh.; Aldalal, Z.

    2003-10-01

    There is probability of exposure to uranium for workers in the phosphoric acid purification (internal exposure) by inhalation, and the deposition of this uranium in organs and tissues, and the consequence excreation out of the body by perspiration or urine. This study focuses on the determination of uranium in urine samples of workers. All results seem to be under the detection limit of the method, therefore no routine monitoring is required. (author)

  9. Cytotoxicity Test Based on Human Cells Labeled with Fluorescent Proteins: Fluorimetry, Photography, and Scanning for High-Throughput Assay.

    Science.gov (United States)

    Kalinina, Marina A; Skvortsov, Dmitry A; Rubtsova, Maria P; Komarova, Ekaterina S; Dontsova, Olga A

    2018-06-01

    High- and medium-throughput assays are now routine methods for drug screening and toxicology investigations on mammalian cells. However, a simple and cost-effective analysis of cytotoxicity that can be carried out with commonly used laboratory equipment is still required. The developed cytotoxicity assays are based on human cell lines stably expressing eGFP, tdTomato, mCherry, or Katushka2S fluorescent proteins. Red fluorescent proteins exhibit a higher signal-to-noise ratio, due to less interference by medium autofluorescence, in comparison to green fluorescent protein. Measurements have been performed on a fluorescence scanner, a plate fluorimeter, and a camera photodocumentation system. For a 96-well plate assay, the sensitivity per well and the measurement duration were 250 cells and 15 min for the scanner, 500 cells and 2 min for the plate fluorimeter, and 1000 cells and less than 1 min for the camera detection. These sensitivities are similar to commonly used MTT (tetrazolium dye) assays. The used scanner and the camera had not been previously applied for cytotoxicity evaluation. An image processing scheme for the high-resolution scanner is proposed that significantly diminishes the number of control wells, even for a library containing fluorescent substances. The suggested cytotoxicity assay has been verified by measurements of the cytotoxicity of several well-known cytotoxic drugs and further applied to test a set of novel bacteriotoxic compounds in a medium-throughput format. The fluorescent signal of living cells is detected without disturbing them and adding any reagents, thus allowing to investigate time-dependent cytotoxicity effects on the same sample of cells. A fast, simple and cost-effective assay is suggested for cytotoxicity evaluation based on mammalian cells expressing fluorescent proteins and commonly used laboratory equipment.

  10. Second-order advantage with excitation–emission photoinduced fluorimetry for the determination of the antiepileptic carbamazepine in environmental waters

    International Nuclear Information System (INIS)

    Lozano, Valeria A.; Escandar, Graciela M.

    2013-01-01

    Graphical abstract: -- Highlights: •A simple and safe method for the emerging contaminant carbamazepine is developed. •MCR-ALS algorithm allows us the quantification in very interfering media. •Determination is accomplished in natural waters using green-chemistry principles. -- Abstract: A photochemically induced fluorescence system combined with second-order chemometric analysis for the determination of the anticonvulsant carbamazepine (CBZ) is presented. CBZ is a widely used drug for the treatment of epilepsy and is included in the group of emerging contaminant present in the aquatic environment. CBZ is not fluorescent in solution but can be converted into a fluorescent compound through a photochemical reaction in a strong acid medium. The determination is carried out by measuring excitation–emission photoinduced fluorescence matrices of the products formed upon ultraviolet light irradiation in a laboratory-constructed reactor constituted by two simple 4 W germicidal tubes. Working conditions related to both the reaction medium and the photoreactor geometry are optimized by an experimental design. The developed approach enabled the determination of CBZ at trace levels without the necessity of applying separation steps, and in the presence of uncalibrated interferences which also display photoinduced fluorescence and may be potentially present in the investigated samples. Different second-order algorithms were tested and successful resolution was achieved using multivariate curve resolution-alternating least-squares (MCR-ALS). The study is employed for the discussion of the scopes and yields of each of the applied second-order chemometric tools. The quality of the proposed method is probed through the determination of the studied emerging pollutant in both environmental and drinking water samples. After a pre-concentration step on a C18 membrane using 50.0 mL of real water samples, a prediction relative error of 2% and limits of detection and quantification of 0.2 and 0.6 ng mL −1 were respectively obtained

  11. Second-order advantage with excitation–emission photoinduced fluorimetry for the determination of the antiepileptic carbamazepine in environmental waters

    Energy Technology Data Exchange (ETDEWEB)

    Lozano, Valeria A.; Escandar, Graciela M., E-mail: escandar@iquir-conicet.gov.ar

    2013-06-11

    Graphical abstract: -- Highlights: •A simple and safe method for the emerging contaminant carbamazepine is developed. •MCR-ALS algorithm allows us the quantification in very interfering media. •Determination is accomplished in natural waters using green-chemistry principles. -- Abstract: A photochemically induced fluorescence system combined with second-order chemometric analysis for the determination of the anticonvulsant carbamazepine (CBZ) is presented. CBZ is a widely used drug for the treatment of epilepsy and is included in the group of emerging contaminant present in the aquatic environment. CBZ is not fluorescent in solution but can be converted into a fluorescent compound through a photochemical reaction in a strong acid medium. The determination is carried out by measuring excitation–emission photoinduced fluorescence matrices of the products formed upon ultraviolet light irradiation in a laboratory-constructed reactor constituted by two simple 4 W germicidal tubes. Working conditions related to both the reaction medium and the photoreactor geometry are optimized by an experimental design. The developed approach enabled the determination of CBZ at trace levels without the necessity of applying separation steps, and in the presence of uncalibrated interferences which also display photoinduced fluorescence and may be potentially present in the investigated samples. Different second-order algorithms were tested and successful resolution was achieved using multivariate curve resolution-alternating least-squares (MCR-ALS). The study is employed for the discussion of the scopes and yields of each of the applied second-order chemometric tools. The quality of the proposed method is probed through the determination of the studied emerging pollutant in both environmental and drinking water samples. After a pre-concentration step on a C18 membrane using 50.0 mL of real water samples, a prediction relative error of 2% and limits of detection and quantification of 0.2 and 0.6 ng mL{sup −1} were respectively obtained.

  12. Second-order advantage with excitation-emission photoinduced fluorimetry for the determination of the antiepileptic carbamazepine in environmental waters.

    Science.gov (United States)

    Lozano, Valeria A; Escandar, Graciela M

    2013-06-11

    A photochemically induced fluorescence system combined with second-order chemometric analysis for the determination of the anticonvulsant carbamazepine (CBZ) is presented. CBZ is a widely used drug for the treatment of epilepsy and is included in the group of emerging contaminant present in the aquatic environment. CBZ is not fluorescent in solution but can be converted into a fluorescent compound through a photochemical reaction in a strong acid medium. The determination is carried out by measuring excitation-emission photoinduced fluorescence matrices of the products formed upon ultraviolet light irradiation in a laboratory-constructed reactor constituted by two simple 4 W germicidal tubes. Working conditions related to both the reaction medium and the photoreactor geometry are optimized by an experimental design. The developed approach enabled the determination of CBZ at trace levels without the necessity of applying separation steps, and in the presence of uncalibrated interferences which also display photoinduced fluorescence and may be potentially present in the investigated samples. Different second-order algorithms were tested and successful resolution was achieved using multivariate curve resolution-alternating least-squares (MCR-ALS). The study is employed for the discussion of the scopes and yields of each of the applied second-order chemometric tools. The quality of the proposed method is probed through the determination of the studied emerging pollutant in both environmental and drinking water samples. After a pre-concentration step on a C18 membrane using 50.0 mL of real water samples, a prediction relative error of 2% and limits of detection and quantification of 0.2 and 0.6 ng mL(-1) were respectively obtained. Copyright © 2013 Elsevier B.V. All rights reserved.

  13. Sensitivity improvement of an immuno-detection method for azaspiracids based on the use of microspheres coupled to a flow-fluorimetry system

    Directory of Open Access Journals (Sweden)

    María Fraga Corral

    2014-06-01

    These results demonstrate the high capability in terms of sensitivity of the microsphere-based immuno-detection assay for AZAs. The immobilization of AZA-1 instead of the synthetic AZA-2 used in Rodríguez et al (Rodriguez et al., 2014, combined with a lower mAb 8F4 concentration provided a remarkable improvement of sensitivity. The ON protocol used in Rodríguez et al. (Rodriguez et al., 2014 displayed a similar IC50 than the new short assay (around 1 nM while the new ON protocol provided an IC50 5-fold more sensitive (0.3 nM. Therefore, the new short assay allows a reduction of the experimental time. Additionally, the increase of sensitivity could help to avoid shellfish matrix interferences. Previously published works using immunoassays for the detection of phycotoxins present in shellfish avoided matrix interference by further extract dilution in combination with an increase of assay sensitivity (Fraga et al., 2012;Fraga et al., 2013. The extraction protocol described by Rodríguez et al. (Rodriguez et al., 2014 will probably be suitable for this newly optimized AZA-detection method since many reagents are the same and the higher sensitivity will allow higher extract dilution. Considering the extraction protocol recovery, sensitivity of the current assay and the regulated limit, shellfish extracts could be diluted up to 1:30 or 1:150 (v/v for detection with the short or long protocols, respectively. Additionally, mAb 8F4 was demonstrated to recognize AZA-2 and AZA-3 with cross-reactivities of 42 and 138 %, respectively. Presumably, this optimized assay will detect these analogs with similar cross-reactivity. The sensitivity of the microsphere-based assay for AZAs is enough to detect these compounds at the regulated levels in shellfish. This microsphere-based multi-detection method provides an easy-to-perform, highly sensitive and rapid method for the detection of AZAs. It could be included in a multi-detection method, which would allow time and sample volume saving due to the simultaneous detection of different toxin classes. The assay could be used in a single or multiplexed format as a semi-quantitative screening tool useful to reduce the number of analysis to be performed by confirmatory, quantitative methods.

  14. In-situ fluorimetry: A powerful non-invasive diagnostic technique for natural dyes used in artefacts. Part II. Identification of orcein and indigo in Renaissance tapestries

    Science.gov (United States)

    Clementi, C.; Miliani, C.; Romani, A.; Santamaria, U.; Morresi, F.; Mlynarska, K.; Favaro, G.

    2009-01-01

    In this paper, three Renaissance tapestries depicting scenes painted by Raffaello Sanzio, conserved at the Vatican Museum, were investigated using in-situ UV-Visible fluorimetric measurements. The results show that this technique is suitable for the detection of natural organic colorants used for dyeing the threads woven in these tapestries. The emission signals detected on red-purple colours were assigned to the colorant orcein and those on different nuances of blue and green colours to indigo by comparison with data from reference laboratory samples. The assignments were supported by chromatographic experiments carried out on threads taken from the back side of the tapestry in the same points analysed by spectrofluorimentry.

  15. The value of fluorimetry (Qubit) and spectrophotometry (NanoDrop) in the quantification of cell-free DNA (cfDNA) in malignant melanoma and prostate cancer patients.

    Science.gov (United States)

    Ponti, Giovanni; Maccaferri, Monia; Manfredini, Marco; Kaleci, Shaniko; Mandrioli, Mauro; Pellacani, Giovanni; Ozben, Tomris; Depenni, Roberta; Bianchi, Giampaolo; Pirola, Giacomo Maria; Tomasi, Aldo

    2018-04-01

    Circulating cell-free tumor DNA (cfDNA) is of crucial interest in oncology. cfDNA constitutes a potential prognostic and therapeutic marker for different solid tumors and can be used in the diagnostic and therapeutic management of cancer patients for which nowadays there are no valid laboratory markers. In the present study, the quality and quantity of the cfDNA were assessed by different quantification procedures, in order to identify the potential applications of these techniques in the preliminary cfDNA quantification. Qubit with single (ss) and double strand (ds) DNA assay kits, NanoDrop and quantitative Real Time PCR (qPCR), were adopted to assess the cfDNA in the blood samples of 18 melanoma patients, 67 prostate cancer patients and 15 healthy controls. The quantification by NanoDrop (average value 8.48ng/μl, 95% confidence limit (CL)=7.23-9.73), Qubit ssDNA (average value 23.08ng/μl, CL=19.88-26.28), dsDNA (average value 4.32ng/μl, CL=3.52-5.12) assay kits and qPCR (average value 0.39ng/μl, CL=0.31-0.47) revealed differences among the four procedures. Qubit 2.0 ss-DNA kit gave higher cfDNA concentration values for all the samples analyzed. In detail, Qubit ssDNA assay revealed higher sensitivity in the quantification of small amounts of pure ss-DNA and ds-DNA, while NanoDrop allowed the assessment of the purity of cfDNA samples. The NanoDrop and Qubit 2.0 measurements were analyzed in order to define their correlation with qPCR cfDNA assessment, showing good correlation values with the qPCR that should be considered the "gold standard". In our proposal, the sequential combination of NanoDrop and Qubit ssDNA methods should be adopted for a cost-effective preliminary assessment of total circulating cfDNA in melanoma and prostate cancer patients, and only discordant values should undergo qPCR assessment. Copyright © 2018 Elsevier B.V. All rights reserved.

  16. Separation and preconcentration of riboflavin from human plasma using polythionine coated magnetite/hydroxyapatite nanocomposite prior to analysis by surfactant-enhanced fluorimetry

    Science.gov (United States)

    Farzin, Leila; Shamsipur, Mojtaba

    2017-09-01

    The exploration of novel adsorption properties of conductive polymers based on hybridization with biocompatible nanomaterials receives an increasing interest. In this regard, hydroxyapatite (HA) bioceramic is of critical importance mainly owing to its facile synthesis, high surface area, economic and low toxicity in biological environments. In this work, we first prepared and characterized a magnetite/hydroxyapatite (Fe3O4/HA) nanocomposite using the bio-waste chicken eggshell via an attractive green way that involved low cost and irrespective of toxicity. Then, polythionine as a novel class of conductive polymers was in situ coated on the synthesized magnetic bioceramic for the separation and preconcentration of riboflavin (vitamin B2) in human plasma before its fluorimetric determination. Considering the putative role of riboflavin in protecting against cancer and cardiovascular diseases, it is essential to evaluate this vitamin in biological fluids. The described method possesses a linear range of 0.75-262.5 μg L- 1 (R2 = 0.9985) and a detection limit of 0.20 μg L- 1 (signal-to-noise ratio of 3). The relative standard deviations (RSDs) for single-sorbent repeatability and sorbent-to-sorbent reproducibility were less than 4.0% and 7.6% (n = 5), respectively. The respective enrichment factor and extraction recovery of the method found to be 35.7 and 98.4%. The analytical performance of method for riboflavin was characterized by good consistency of the results with those obtained by the enzyme-linked immunosorbent assay (ELISA) conventional method (p-value of < 0.05). The optimized protocol intended for control determinations of riboflavin in human subjects and is addressed to clinical laboratories.

  17. X-ray fluorimetry in tests of the aerosol substances. [In the atmosphere]. Rentgenowska analiza fluorescencyjna w badaniach skladu pierwiastkowego aerozoli w atmosferze

    Energy Technology Data Exchange (ETDEWEB)

    Walentek, A.; Cudny, W.; Marzec, J.; Pawlowski, Z.; Zaremba, K. (Politechnika Warszawska, Warsaw (Poland). Inst. Radioelektroniki)

    1992-01-01

    Test results of aerosol samples from atmospheric air collected from membrane drains were presented. The ANFLU-2 analyzer constructed at the Warsaw Technical University were used for testing. (author). 3 figs, 1 tab.

  18. Establishing the anion recognition correlation of the 2-(2-methoxyphenyl)-1H-imidazo [4, 5-f][1,10] phenanthroline and its Ru(bipy){sub 2}{sup 2+} complex via fluorimetry

    Energy Technology Data Exchange (ETDEWEB)

    Alreja, Priya; Kaur, Navneet, E-mail: neet_chem@yahoo.co.in

    2016-11-15

    2-(2-methoxyphenyl)-1H-imidazo [4, 5-f][1, 10] phenanthroline (1) and its Ru(bipy){sub 2}{sup 2+} complex (2) were synthesized and the anion sensing properties of the two were well evaluated and compared by fluorescence titration experiments. Both 1 and 2 were fluorescent in DMSO solution and possessed an anion binding imidazole N–H site to interact with the anions. 1 did not exhibit any fluorescent changes with even most basic anions in DMSO solution but on introduction of Ru metal center via synthesis of its Ru(bipy){sub 2}{sup 2+} complex (2), the N–H acidity enhanced appreciably and it could detect acetate even at very low concentrations (LOD=5.25×10{sup −7}). The fluorescence intensity of 2 was almost completely quenched and its fluorescent red orange color disappeared with the addition of acetate ions. The spatial arrangement of receptor 2 fits well for the triangular shape of AcO{sup −} ion, hence making it the most preferred anion for binding. Both {sup 1}H NMR titrations and DFT computational calculations supported the deprotonation mechanism.

  19. Fluorescence of UO22+ in different acidic media containing cationic and anionic impurities. Application to the elaboration of a very sensitive dosing method of Uranium in solution by fluorimetry and to the study of the kinetics of U-6 reduction by Iron

    International Nuclear Information System (INIS)

    Belkadi, L.

    1990-09-01

    The use of the fluorimetric analysis method in phosphoric medium proved that this method is very sensitive for detecting Uranium traces (10 E-10 M). The dosing can be carried out after a simple calibration of the device and without calling for the addition techniques. The interference of most organic matters is eliminated by the 337 nm exciting radiation. The inhibition of the fluorescence induced by anions and cations is generally resolved by a simple dilution. The nitrates that have a harmful effect on the Uranium fluorescence have been eliminated by successive evaporations. This method, as it has been improved in this work, is applied to the study of U-6 reduction by metallic Iron and Fe-2 in orthophosphoric acid medium in case the absorption spectrophotometry becomes inoperative. 37 figs., 14 tabs., 50 refs. (author)

  20. Quality engineering and control semiannual progress report November and December 1976 and January--April 1977

    International Nuclear Information System (INIS)

    Carpenter, R.L.

    1977-01-01

    Research and development activities are reported on absorption spectroscopy, calorimetry, electrolysis, emission spectroscopy, fluorimetry gas chromatography, infrared spectroscopy, liquid chromatography, liquid scintillation counting, mass spectroscopy, microscopy, radiometric analysis, and thermal analysis. A group of miscellaneous projects are also described

  1. SHPOLSKII FLUOROMETRIC-DETERMINATION OF POLYCYCLIC AROMATIC-HYDROCARBONS IN COMPLEX ENVIRONMENTAL-SAMPLES

    NARCIS (Netherlands)

    ARIESE, F; GOOIJER, C; Velthorst, N.H.; Hofstraat, J.W.

    1991-01-01

    High-resolution fluorimetry in low-temperature n-alkane Shpol'skii matrices is a powerful technique for the analysis of rigid, non-polar compounds like polycyclic aromatic hydrocarbons. Because of the method's sensitivity and selectivity, sample clean-up, preconcentration and even chromatographic

  2. SHORT COMMUNICATION SPECTROPHOTOMETRIC ...

    African Journals Online (AJOL)

    Thus, the determination of oxalic acid in biological samples such as tea is significant. At present the methods for the determination of oxalic acid include chemical luminescence [2], fluorimetry [3,4], and permanganimetric titration. [5]. These methods have different extent of defects, such as instrumental operation complexity,.

  3. Geochemistry and analysis of uranium - an overview; with special reference to geochemical exploration in Malaysia

    Energy Technology Data Exchange (ETDEWEB)

    Choong, W Y; Seevaratnam, S

    1981-01-01

    The paper presents discussions of the general chemistry of uranium, its abundance and occurrence, and the geochemistry applications to exploration of uranium in Malaysia in silts vs. water. Methods of analysis at the Geological Survey of Malaysia are fluorimetry for uranium analysis of ores and minerals.

  4. Recent applications of lasers in analytical chemistry

    International Nuclear Information System (INIS)

    Lytle, F.E.

    1984-01-01

    This review primarily will update two recent articles concerning the use of lasers in analytical fluorimetry. The discussion will focus on the use of instrumental techniques developed to improve the working detection limit via increases in selectivity. Examples will include chromatography, time resolution and line narrowing. In addition, the topic of multiphoton ionization/mass spectrometry will be covered. 18 refs., 1 tab

  5. Introduction to the problems of uranium analysis

    International Nuclear Information System (INIS)

    Suschny, O.

    1983-08-01

    Brief information is given on different techniques used for recognition, separation and determination of uranium. The analytical methods dealt with are: radiometry, neutron activation analysis, delayed neutron counting, X-ray fluorescence analysis, fluorimetry, spectral photometry, gravimetry, and volumetry. Selection of a suitable method for determining uranium in geochemical surveys is also discussed

  6. Optimization of the crystallizability of a single-chain antibody fragment

    Czech Academy of Sciences Publication Activity Database

    Škerlová, Jana; Král, Vlastimil; Fábry, Milan; Sedláček, Juraj; Veverka, Václav; Řezáčová, Pavlína

    2014-01-01

    Roč. 70, č. 12 (2014), s. 1701-1706 ISSN 1744-3091 R&D Projects: GA MŠk(CZ) LK11205 Institutional support: RVO:61388963 ; RVO:68378050 Keywords : single-chain antibody fragment * Thermofluor assay * differential scanning fluorimetry * crystallizability optimization * oligomerization * crystallization Subject RIV: CE - Biochemistry Impact factor: 0.527, year: 2014

  7. A method for the quantitative determination of uranium-233 in an irradiated thorium rod; Une methode de dosage de l'uranium 233 contenu dans un barreau de thorium irradie

    Energy Technology Data Exchange (ETDEWEB)

    Bathellier, A; Sontag, R; Chesne, A [Commissariat a l' Energie Atomique, Saclay (France).Centre d' Etudes Nucleaires

    1961-07-01

    A rapid method for the quantitative determination of uranium-233 in irradiated thorium is described. A 30 per cent solution of trilaurylamine in xylene is used to extract the uranium from an aqueous hydrochloric acid solution and separate it from the thorium. This may be followed by {alpha} counting or fluorimetry. The practical operating conditions of the separation are discussed in detail. (author) [French] Une methode rapide de dosage de l'uranium-233 contenu dans le thorium irradie est decrite. Elle utilise la trilauryfamine a 30 pour cent dans le xylene pour extraire l'uranium d'une dissolution aqueuse chlorhydrique et le separer du thorium. Le comptage {alpha} ou la fluorimetrie sont alors possibles. Les conditions operatoires de la separation sont discutees et precisees. (auteur)

  8. Measurements of natural uranium concentration in Caspian Sea and Persian Gulf water by laser fluorimetric method

    International Nuclear Information System (INIS)

    Garshasbi, H.; Karimi Diba, J.; Jahanbakhshian, M. H.; Asghari, S. K.; Heravi, G. H.

    2005-01-01

    Natural uranium exists in earth crust and seawater. The concentration of uranium might increase by human manipulation or geological changes. The aim of this study was to verify susceptibility of laser fluorimetry method to determine the uranium concentration in Caspian Sea and Persian Gulf water. Materials and Methods: Laser fluorimetric method was used to determine the uranium concentration in several samples prepared from Caspian Sea and Persian Gulf water. Biological and chemical substances were eliminated in samples for better evaluation of the method. Results: As the concentration of natural uranium in samples increases, the response of instrument (uranium analyzer) increases accordingly. The standard deviation also increased slightly and gradually. Conclusion: Results indicate that the laser fluorimetry method show a reliable and accurate response with uranium concentration up to 100 μg/L in samples after removal of biological and organic substances

  9. Studies on preparation of some alkaloid complexes and solvent extraction

    Energy Technology Data Exchange (ETDEWEB)

    Mohamed, A A

    1990-12-31

    Describes the various methods available in the literature for the determination of different classes of alkaloids spectrochemical (e.g.spectrophotometry and colorimetry, fluorimetry and phosphorimetry, nephelometry and reiractometry), electrochemical (e.g. polarography, amperometry, conductometry, coulometry and potentiometry), volumetric (e.q. acid-base, iodometry , bromimetry , oxidimetry, complexometry and precipitation ) and miscellaneous methods (e.g.gasliquid chromatography, gravimetry, thermometry and rediometry ) has been discussed. 31 tabs; 29 figs; 402 refs.

  10. Studies on preparation of some alkaloid complexes and solvent extraction

    International Nuclear Information System (INIS)

    Mohamed, A.A.

    1989-01-01

    Describes the various methods available in the literature for the determination of different classes of alkaloids spectrochemical (e.g.spectrophotometry and colorimetry, fluorimetry and phosphorimetry, nephelometry and reiractometry), electrochemical (e.g. polarography, amperometry, conductometry, coulometry and potentiometry), volumetric (e.q. acid-base, iodometry , bromimetry , oxidimetry, complexometry and precipitation ) and miscellaneous methods (e.g.gasliquid chromatography, gravimetry, thermometry and rediometry ) has been discussed

  11. Study of performance characteristics of a radiochemical method to determine uranium in biological samples

    International Nuclear Information System (INIS)

    Puga, Maria J.; Cerchietti, Maria L.R.; Prudenzo, J.E.; Arguelles, Maria G.

    2005-01-01

    In this paper is described a methodology to calculate detection limit (Ld), quantification level (Lq) and minimum detectable activity (MDA) in a radiochemical method for determination of uranium in urine samples. The concentration is measured by fluorimetry and alpha gross activity using liquid scintillation counting (LSC). The calculation of total propagated uncertainty on a spike sample is presented. Furthermore, the major sources of uncertainty and percentage contribution in both measurements are assessed. (author)

  12. Application of the multi-element analysis by X-fluorescence and neutron activation to the characterization of an archaeological site

    International Nuclear Information System (INIS)

    Rossini, I.

    1991-06-01

    The first part of this thesis is about possible analysis methods (XRF, PIXE, INAA, laser fluorimetry, and ICP), applied to Uranium, Thorium and Rubidium assays in archaeological clays and potteries. The best results have been obtained with Neutron Activation technics. The second part is about the multi-element analysis of quarries and about the research by statistical treatment of correlations between the element concentrations and the sampling sites (excavations, quarries)

  13. Des réactions photochimiques aux interfaces atmosphériques

    OpenAIRE

    Tinel , Liselotte

    2015-01-01

    The works presented in this thesis concern firstly the characterization of two new photosensitizers by spectroscopic methods. This way the kinetics of the oxidation reaction between the triplet state of the photosensitizers, imidazole-2-carboxaldehyde and 6-carboxypterin, and three halides have been determined by laser flash photolysis. Also, the reactivity of the singlet state of 6-carboxypterin with halides and four organic acids has been studied by static fluorimetry. These photosensitizer...

  14. Uranium determination in water

    International Nuclear Information System (INIS)

    Prudenzo, E.J.; Puga, Maria J.; Cerchietti, Maria L.R.; Arguelles, Maria G.

    2005-01-01

    In our laboratory, a procedure has been assessed to determine uranium content of water in normal situations. The method proposed without sample pre-treatment, is simple and rapid. Uranium mass is measured by fluorimetry. For calculation of detection limit (Ld) and quantification level (Lq) we used blank samples and the results were analyzed for different statistical test. The calculation of total propagated uncertainty and sources contribution on real samples are presented. (author)

  15. Application of fission track technique for estimation of uranium concentration in drinking waters of Punjab

    International Nuclear Information System (INIS)

    Prabhu, S.P.; Raj, Sanu S.; Sawant, P.D.; Kumar, Ajay; Sarkar, P.K.; Tripathi, R.M.

    2010-01-01

    Full text: Drinking water samples were collected from four different districts, namely Bhatinda, Mansa, Faridkot and Firozpur, of Punjab for ascertaining the U(nat.) concentrations. The samples were collected from bore wells, hand pumps, tube wells and treated municipal water supply. All these samples (235 nos.) collected were preserved and processed by following the international standard protocol and analyzed by Laser Fluorimetry. Results of analysis by laser fluorimetry have been already reported. To ensure accuracy of the data obtained by laser fluorimetry, few samples (10 nos) from each district were analyzed by alpha spectrometry as well as by fission track analysis (FTA) technique. FTA in solution media for uranium has been already standardized in Bioassay laboratory of Health Physics Division. Few of drinking water sample was directly transferred to polythene tube sealed at one end. Lexan detector with proper identification mark was immersed in the samples and the other open end of the tube was also heat-sealed. Two tubes containing samples and one containing uranium standard (80 ppb) were irradiated in the Pneumatic Carrier Facility (PCF) of DHRUVA reactor. The Lexan detectors were then chemically etched and tracks were counted under an optical microscope at 400X magnification. Concentration of uranium in sample was determined by comparison technique. Quality assurance was carried out by replicate analysis and by analysis of standard reference materials. Uranium concentration in these samples ranged from 3.2 to 60.5 ppb with an average of 28.8 ppb. A t-test analysis for paired data was done to compare the results obtained by FTA and those obtained by laser fluorimeter. The calculated value for t is -1.19, which is greater than the tabulated value of t for 40 observations (-2.02 at 95% confidence level). This shows that the results of the measurements carried out by the FTA and laser fluorimetry are not significantly different. The preliminary studies

  16. Protective effect of Clostridium tyrobutyricum in acute dextran sodium sulphate-induced colitis: differential regulation of tumour necrosis factor-α and interleukin-18 in BALB/c and severe combined immunodeficiency mice

    Czech Academy of Sciences Publication Activity Database

    Hudcovic, Tomáš; Kolínská, Jiřina; Klepetář, Jan; Štěpánková, Renata; Řezanka, Tomáš; Šrůtková, Dagmar; Schwarzer, Martin; Erban, V.; Du, Zhengyu; Wells, J. M.; Hrnčíř, Tomáš; Tlaskalová-Hogenová, Helena; Kozáková, Hana

    2012-01-01

    Roč. 167, č. 2 (2012), s. 356-365 ISSN 0009-9104 R&D Projects: GA ČR GA303/08/0367; GA ČR GA303/09/0449; GA ČR(CZ) GAP304/11/1252; GA MŠk 2B06155; GA MŠk(CZ) ME10017 Institutional research plan: CEZ:AV0Z50200510; CEZ:AV0Z50110509 Keywords : butyrate * Clostridium tyrobutyricum * confocal fluorimetry Subject RIV: EC - Immunology Impact factor: 3.409, year: 2012

  17. Internal contamination by natural uranium: monitoring by analysis of urine of individuals exposed by occupational inhalation

    International Nuclear Information System (INIS)

    Ramalho, A.T.

    1982-01-01

    Urine samples from men working at Usina Santo Amaro (USAM - State of Sao Paulo), a monazite refinery, were analysed for uranium concentration, using fluorometric analysis and alpha spectrometry. All samples analysed presented uranium concentration below the lower limit of detection. Theoretical values were calculated for uranium concentration in urine samples from workers at the annual limit of intake (ALI) for inhalation of natural uranium, recommended in Publication 30 of the International Commission on Radiological Protection (ICRP, 1979). The two different methods used for analysis of natural uranium concentration in the urine samples were compared: fluorimetry and alpha spectrometry. (author)

  18. Geophysical characterization from Itu intrusive suite

    International Nuclear Information System (INIS)

    Pascholati, M.E.

    1989-01-01

    The integrated use of geophysical, geological, geochemical, petrographical and remote sensing data resulted in a substantial increase in the knowledge of the Itu Intrusive Suite. The main geophysical method was gamma-ray spectrometry together with fluorimetry and autoradiography. Three methods were used for calculation of laboratory gamma-ray spectrometry data. For U, the regression method was the best one. For K and Th, equations system and absolute calibration presented the best results. Surface gamma-ray spectrometry allowed comparison with laboratory data and permitted important contribution to the study of environmental radiation. (author)

  19. Synthesis, spectroscopic and thermochemical studies of some novel carbazole-pyridine co-polymers (abstract)

    International Nuclear Information System (INIS)

    Saeed, A.; Irfan, M.

    2011-01-01

    Two series of a novel class of carbazole-4-phenylpyridine co-polymers has been synthesized by a modified Chichibabin reaction. The synthesis was carried out by a simple and cheaper method compared to the lengthy methods usually adopted for the preparation of carbazole-pyridine copolymers which involve costly catalysts. All the polymers were characterized by IR, /sup 1/H and /sup 13/C NMR, UV-vis spectroscopy, fluorimetry, TGA and DSC. The weight average molecular masses (M/sub w/) of the polymers were estimated by Laser Light Scattering (LLS) technique. (author)

  20. Remote sensing of chlorophyll a fluorescence of vegetation canopies. 1. Near and far field measurement techniques

    International Nuclear Information System (INIS)

    Cecchi, G.; Mazzinghi, P.; Pantani, L.; Valentini, R.; Tirelli, D.; De Angelis, P.

    1994-01-01

    This article presents instruments and techniques, used in several vegetation monitoring experiments. Simultaneous monitoring was performed with different approaches, including fluorescence lidar and passive remote sensing, leaf level reflectance, and laser fluorimetry, and compared with physiological measurements. Most of the instrumentation described was designed and built for this application. Experiments were carried out in the laboratory and in the field, to investigate the relationship between chlorophyll fluorescence spectra and plant ecophysiology. Remote sensing, spectroscopy, and ecophysiology data were then collected by an intensive research team, joining different experiences and working in national and international projects

  1. Assessment of radiation dose due to intake of uranium through ground water and its carcinogenic and non-carcinogenic risks in SW and NE Punjab

    International Nuclear Information System (INIS)

    Bajwa, B.S.

    2017-01-01

    In the present investigations, Laser Fluorimetry technique has been used for the microanalysis of uranium content in drinking water samples collected from different sources like the hand pumps, tube wells of various depths from wide range of locations in the four districts of SW-Punjab, India and two districts of NE - Punjab, India. The purpose of this study was to investigate the uranium concentration levels of ground water being used for drinking and to determine its health effects, if any, to the local population of these regions

  2. Effect of in vivo γ-irradiation on the binding of wheat germ agglutinin on lymphocyte plasma membranes

    International Nuclear Information System (INIS)

    Moullier, P.; Daveloose, D.; Dubos, M.; Leterrier, F.; Hoebeke, J.

    1986-01-01

    Using quantitative fluorimetry with fluoresceinated wheat germ agglutinin, we have been able to investigate in vivo gamma radiation-induced damage at the outer membrane level of rat splenic lymphocytes, namely damage to the glucosidic moieties of membrane glycoproteins and glycolipids. This paper demonstrates that below an irradiation level of 1 gray (Gy), removal of sialic acid is the major feature leading to new exposed specific binding sites for wheat germ agglutinin, since this lectin is specific for sialic acid and N-acetyl-D-glucosamine. Our studies also suggest that above 1 Gy of irradiation more internal damage occurs, since we observed a striking decrease in wheat germ agglutinin binding sites. (orig.)

  3. Estudo das dispersões aquosas de nanotubos de carbono utilizando diferentes surfactantes Study of aqueous dispersions of carbon nanotubes using different surfactants

    Directory of Open Access Journals (Sweden)

    Isabella R. da Silva

    2013-01-01

    Full Text Available The dispersion of carbon nanotubes in water for their utilization in nanoscale devices is a challenging task. Comparative studies on interaction and dispersion of multi-wall carbon nanotubes (MWNT using two different surfactants (sodium dodecyl sulfate, SDS, and polyoxyethylenesorbitanmonooleate, Tween 80 are presented. The interaction between carbon nanotubes and surfactants was studied by tensiometry, conductivimetry, and fluorimetry. The dispersions of MWNT in surfactants were characterized using a UV-vis spectrophotometer. For effective dispersion, the minimum weight ratio of MWNT to surfactant was 1:41 and 1:3 for SDS and Tween 80, respectively.

  4. Estudo das dispersões aquosas de nanotubos de carbono utilizando diferentes surfactantes

    Directory of Open Access Journals (Sweden)

    Isabella R. da Silva

    2013-01-01

    Full Text Available The dispersion of carbon nanotubes in water for their utilization in nanoscale devices is a challenging task. Comparative studies on interaction and dispersion of multi-wall carbon nanotubes (MWNT using two different surfactants (sodium dodecyl sulfate, SDS, and polyoxyethylenesorbitanmonooleate, Tween 80 are presented. The interaction between carbon nanotubes and surfactants was studied by tensiometry, conductivimetry, and fluorimetry. The dispersions of MWNT in surfactants were characterized using a UV-vis spectrophotometer. For effective dispersion, the minimum weight ratio of MWNT to surfactant was 1:41 and 1:3 for SDS and Tween 80, respectively.

  5. Applications of optical fiber to remote laser fluorescence analysis

    International Nuclear Information System (INIS)

    Kim, Cheol Jung; Shin, Jang Soo; Lee, Sang Mock; Kim, Jeong Moog; Kim, Duk Heon; Hong, Seok Kyung

    1991-12-01

    Fluorescence analysis using time-resolved laser fluorimetry has been used for trace uranium analysis because this method shows high sensitivity and low detection limit and is less matrix dependent than any other fluorimetric measurement. By this time, the uranium analyses in the solution of reprocessing process or high radioactive area have been primarily analyzed by sampling of the solution, but recently, a study on a remote uranium fluorescence analysis using optical fiber has been setting out based on the development of an optical fiber with radiation resistivity and of an advanced laser excitation source. Laser fluorimetry developed by our laboratory for trace uranium analyses in uranium handling process or in urine samples of workers in a nuclear facility has been used in our institute since 1988. A development of the system for remote control of uranium fluorescence analysis will be expected to contribute to an on-line uranium concentration monitoring in the cooling water of reconversion stream. In this report, we summarize the information related to fluorescence analyses and remote fluorescence monitoring methods established by foreign countries and our laboratory. We also present a future research direction for remote on-line monitoring of uranium in conversion or reconversion process. (Author)

  6. Analysis of Uranium and Thorium in Radioactive Wastes from Nuclear Fuel Cycle Process

    International Nuclear Information System (INIS)

    Gunandjar

    2008-01-01

    The assessment of analysis method for uranium and thorium in radioactive wastes generated from nuclear fuel cycle process have been carried out. The uranium and thorium analysis methods in the assessment are consist of Titrimetry, UV-VIS Spectrophotometry, Fluorimetry, HPLC, Polarography, Emission Spectrograph, XRF, AAS, Alpha Spectrometry and Mass Spectrometry methods. From the assessment can be concluded that the analysis methods of uranium and thorium content in radioactive waste for low concentration level using UV-VIS Spectrometry is better than Titrimetry method. While for very low concentration level in part per billion (ppb) can be used by Neutron Activation Analysis (NAA), Alpha Spectrometry and Mass Spectrometry. Laser Fluorimetry is the best method of uranium analysis for very low concentration level. Alpha Spectrometry and ICP-MS (Inductively Coupled Plasma Mass Spectrometry) methods for isotopic analysis are favourable in the precision and accuracy aspects. Comparison of the ICP-MS and Alpha Spectrometry methods shows that the both of methods have capability to determining of uranium and thorium isotopes content in the waste samples with results comparable very well, but the time of its analysis using ICP-MS method is faster than the Alpha Spectrometry, and also the cost of analysis for ICP-MS method is cheaper. NAA method can also be used to analyze the uranium and thorium isotopes, but this method needs the reactor facility and also the time of its analysis is very long. (author)

  7. Radioactivity Content in Phosphoric Acid Used for Fertilizer Production

    International Nuclear Information System (INIS)

    Ibrahiem, N.M.; Hamed, A.A.

    2003-01-01

    Uranium content in phosphoric acid used fertilizer production was measured by alpha spectrometry, laser fluorimetry high resolution gamma spectrometry. Also, polonium-210 content was determined in phosphoric acid by alpha spectrometry. Uranium-234 and uranium-238 concentrations, measured by alpha spectroscopy, were found to be 601 and 507 Bq I -1 , respectively. Total uranium content obtained by laser fluorimetry was about 545 BqI - (45.4ppm). Gamma spectroscopy analysis gave the concentrations of 40 K, 238 U, 235 U, 214 Pb, 214 Bi and 208 TI, as 17,644,19.5, 1.2,1.3 and 9.4 Bq I -1 , respectively. Polonium-210 concentration was found to be about 3.1 Bq I -1 . Uranium-232 and polonium-208 were used as yield tracers, for alpha measurements of uranium and polonium, respectively. Samples of the tri-super phosphate (TSP) and single-super phosphate (SSP) fertilizers and the phosphogypsum produced were also analyzed by gamma spectroscopy. Uranium content in both phosphate fertilizers was 3205 and 1440 Bq Kg -1 for 238 U and 83 and 35 Bq Kg -1 for, 235 U respectively

  8. U(VI) and Eu(III) ion sorption in the interface solution-phosphate solids: Structural study and mechanisms; Sorption des ions U(VI) et Eu(III) a l`interface solution - solides phosphates: Etude structurale et mechanismes

    Energy Technology Data Exchange (ETDEWEB)

    Drot, Romuald [Inst. de Physique Nucleaire, Paris-11 Univ., 91 - Orsay (France)

    1998-09-18

    As part of the storage of nuclear wastes in a deep underground disposal, radionuclides sorption on geological or engineered barriers is one of the most important factor which could enhance retardation. Thus, the knowledge of such mechanisms is needed. For this purpose, we chose to experimentally define sorption equilibria before performing simulation of retention data. Several phosphate compounds are potential candidates as engineered barrier additives. We considered Th{sub 4}(PO{sub 4}){sub 4}P{sub 2}O{sub 7}, Zr{sub 2}O(PO{sub 4}){sub 2} which allow to study the effect of PO{sub 4} and P{sub 2}O{sub 7} groups separately. Eu(III) and U(IV) ions were used as structural probes in order to simulate actinides (III) and (VI) behavior. X-ray powder diffraction, IR spectroscopy and electron probe microanalysis were used to characterized the synthesized solids. Electrophoretic measurements showed an amphoteric behavior of surface sites. Moreover, laser spectro-fluorimetry experiments indicated that no diffusion phenomena of the sorbed ion inside the solid occurs. Thus, we considered that a surface complexation model should be applied. Laser spectro-fluorimetry and XPS allowed to determine the nature of surface sites. ZrP{sub 2}O{sub 7} presents only one single site (P{sub 2}O{sub 7} groups) whereas Th{sub 4}(PO{sub 4}){sub 4}P{sub 2}O{sub 7} and Zr{sub 2}O(PO{sub 4}){sub 2} admit two types of sites (PO{sub 4}/P{sub 2}O{sub 7} and PO{sub 4}/oxo groups, respectively). Sorbed species were identified using laser spectro-fluorimetry which indicate that, in KNO{sub 3} 0.5 M medium and for a known surface site, there are two surface complexes for U(VI) (sorption of UO{sup 2+}{sub 2} et de UO{sub 2}NO{sup +}{sub 3} species) and only one for Eu(III) (sorption of EuNO{sup 2+}{sub 3}). They are linked to the substrate as bidentate inner sphere complexes (EXAFS study). Surface acidity constants were determined by simulation of potentiometric titration curves obtained for each solid

  9. U(VI) and Eu(III) ion sorption in the interface solution-phosphate solids: Structural study and mechanisms

    International Nuclear Information System (INIS)

    Drot, Romuald

    1998-01-01

    As part of the storage of nuclear wastes in a deep underground disposal, radionuclides sorption on geological or engineered barriers is one of the most important factor which could enhance retardation. Thus, the knowledge of such mechanisms is needed. For this purpose, we chose to experimentally define sorption equilibria before performing simulation of retention data. Several phosphate compounds are potential candidates as engineered barrier additives. We considered Th 4 (PO 4 ) 4 P 2 O 7 , Zr 2 O(PO 4 ) 2 which allow to study the effect of PO 4 and P 2 O 7 groups separately. Eu(III) and U(IV) ions were used as structural probes in order to simulate actinides (III) and (VI) behavior. X-ray powder diffraction, IR spectroscopy and electron probe microanalysis were used to characterized the synthesized solids. Electrophoretic measurements showed an amphoteric behavior of surface sites. Moreover, laser spectro-fluorimetry experiments indicated that no diffusion phenomena of the sorbed ion inside the solid occurs. Thus, we considered that a surface complexation model should be applied. Laser spectro-fluorimetry and XPS allowed to determine the nature of surface sites. ZrP 2 O 7 presents only one single site (P 2 O 7 groups) whereas Th 4 (PO 4 ) 4 P 2 O 7 and Zr 2 O(PO 4 ) 2 admit two types of sites (PO 4 /P 2 O 7 and PO 4 /oxo groups, respectively). Sorbed species were identified using laser spectro-fluorimetry which indicate that, in KNO 3 0.5 M medium and for a known surface site, there are two surface complexes for U(VI) (sorption of UO 2+ 2 et de UO 2 NO + 3 species) and only one for Eu(III) (sorption of EuNO 2+ 3 ). They are linked to the substrate as bidentate inner sphere complexes (EXAFS study). Surface acidity constants were determined by simulation of potentiometric titration curves obtained for each solid suspension using FITEQL code (CCM). As sorption equilibria were defined, experimental retention data simulation was performed with respect to structural

  10. 340 nm pulsed UV LED system for europium-based time-resolved fluorescence detection of immunoassays

    DEFF Research Database (Denmark)

    Rodenko, Olga; Fodgaard, Henrik; Tidemand-Lichtenberg, Peter

    2016-01-01

    We report on the design, development and investigation of an optical system based on UV light emitting diode (LED) excitation at 340 nm for time-resolved fluorescence detection of immunoassays. The system was tested to measure cardiac marker Troponin I with a concentration of 200 ng....../L in immunoassay. The signal-to-noise ratio was comparable to state-of-the-art Xenon flash lamp based unit with equal excitation energy and without overdriving the LED. We performed a comparative study of the flash lamp and the LED based system and discussed temporal, spatial, and spectral features of the LED...... excitation for time-resolved fluorimetry. Optimization of the suggested key parameters of the LED promises significant increase of the signal-to-noise ratio and hence of the sensitivity of immunoassay systems....

  11. 340 nm pulsed UV LED system for europium-based time-resolved fluorescence detection of immunoassays.

    Science.gov (United States)

    Rodenko, Olga; Fodgaard, Henrik; Tidemand-Lichtenberg, Peter; Petersen, Paul Michael; Pedersen, Christian

    2016-09-19

    We report on the design, development and investigation of an optical system based on UV light emitting diode (LED) excitation at 340 nm for time-resolved fluorescence detection of immunoassays. The system was tested to measure cardiac marker Troponin I with a concentration of 200 ng/L in immunoassay. The signal-to-noise ratio was comparable to state-of-the-art Xenon flash lamp based unit with equal excitation energy and without overdriving the LED. We performed a comparative study of the flash lamp and the LED based system and discussed temporal, spatial, and spectral features of the LED excitation for time-resolved fluorimetry. Optimization of the suggested key parameters of the LED promises significant increase of the signal-to-noise ratio and hence of the sensitivity of immunoassay systems.

  12. Contribution to the study of uranium migration and some trace elements in solution from Pocos de Caldas uranium mining

    International Nuclear Information System (INIS)

    Zenaro, R.

    1989-01-01

    It was studied the chemical composition of ground water from four boreholes as a contribution to the hydrogeochemical studies in the Pocos de Caldas uranium mining. Methods for water analyses were selected and optimized in order to determine the main anions, specially the ones which form stable complexes with uranium ions. Fluoride and chloride were determined by potentiometry; phosphate, nitrate and silicate by spectrophotometry. Cations were determined by atomic absorption spectrophotometry flame emission and argon plasma emission excited by continuous current arch (DCP). Uranium was determined by fluorimetry with a concentration range from 3 to 7 ppb and its distribution calculated among the different species into solution through the measures of pH, Eh, anion amounts and stability of their respective complexes. (author) [pt

  13. Application of fission track technique for estimation of uranium concentration in drinking waters of Punjab

    International Nuclear Information System (INIS)

    Prabhu, S.P.; Sawant, P.D.; Raj, S.S.; Kumar, A.; Sarkar, P.K.; Tripathi, R.M.

    2012-01-01

    Drinking water samples were collected from four different districts, namely Bhatinda, Mansa, Faridkot and Firozpur, of Punjab for ascertaining the U(nat.) concentrations. All samples were preserved, processed and analyzed by laser fluorimetry (LF). To ensure accuracy of the data obtained by LF, few samples (10 nos) from each district were analyzed by alpha spectrometry as well as by fission track analysis (FTA) technique. For FTA technique few μl of water sample was transferred to polythene tube, lexan detector was immersed in it and the other end of the tube was also heat-sealed. Two samples and one uranium standard were irradiated in DHRUVA reactor. Irradiated detectors were chemically etched and tracks counted using an optical microscope. Uranium concentrations in samples ranged from 3.2 to 60.5 ppb and were comparable with those observed by LF. (author)

  14. Mass spectrometry for fragment screening.

    Science.gov (United States)

    Chan, Daniel Shiu-Hin; Whitehouse, Andrew J; Coyne, Anthony G; Abell, Chris

    2017-11-08

    Fragment-based approaches in chemical biology and drug discovery have been widely adopted worldwide in both academia and industry. Fragment hits tend to interact weakly with their targets, necessitating the use of sensitive biophysical techniques to detect their binding. Common fragment screening techniques include differential scanning fluorimetry (DSF) and ligand-observed NMR. Validation and characterization of hits is usually performed using a combination of protein-observed NMR, isothermal titration calorimetry (ITC) and X-ray crystallography. In this context, MS is a relatively underutilized technique in fragment screening for drug discovery. MS-based techniques have the advantage of high sensitivity, low sample consumption and being label-free. This review highlights recent examples of the emerging use of MS-based techniques in fragment screening. © 2017 The Author(s). Published by Portland Press Limited on behalf of the Biochemical Society.

  15. Sensing a Sensor: Identifying the Mechanosensory Function of Primary Cilia

    Science.gov (United States)

    Prasad, Rahul M.; Jin, Xingjian; Nauli, Surya M.

    2014-01-01

    Over the past decade, primary cilia have emerged as the premier means by which cells sense and transduce mechanical stimuli. Primary cilia are sensory organelles that have been shown to be vitally involved in the mechanosensation of urine in the renal nephron, bile in the hepatic biliary system, digestive fluid in the pancreatic duct, dentin in dental pulp, lacunocanalicular fluid in bone and cartilage, and blood in vasculature. The prevalence of primary cilia among mammalian cell types is matched by the tremendously varied disease states caused by both structural and functional defects in cilia. In the process of delineating the mechanisms behind these disease states, calcium fluorimetry has been widely utilized as a means of quantifying ciliary function to both fluid flow and pharmacological agents. In this review, we will discuss the approaches used in associating calcium levels to cilia function. PMID:24839551

  16. Effect of 60 Co gamma radiation on crystalline proteins

    International Nuclear Information System (INIS)

    Bernardes, D.M.L.

    1991-01-01

    In order to study the effects of 6 0 Co gamma radiation on crystalline proteins an in vitro system was set up. For that, aqueous solutions from bovine crystalline were used irradiated with 0, 5.000, 10.000, 15.000, 20.000 and 25.000 Gy. The treatment led to protein alterations determined by different methods. By turbidimetry the formation of aggregates that increased with the radiation dose was revealed. The same observation was done from viscosity data and from the UV spectrum of the samples. From amino acid analysis and fluorimetry determinations, tryptophan appeared as the most sensitive amino acid. An increase in the free-S H-groups was also observed. After the standardization of the method, the radio modifier capability of glutathione, amino ethyl thiourea, mercapto ethyl alanine and dimethyl sulfoxide was tested. The results showed that in the presence of those substances the radiation effect was diminished. (author)

  17. Evaluation of oil and grease removal by adsorptive polymeric resins in semi-industrial scale: influence of temperature; Avaliacao da remocao de oleos e graxas por resinas polimericas adsorventes em escala semi-industrial: influencia da temperatura

    Energy Technology Data Exchange (ETDEWEB)

    Oliveira, Luis F.S. de; Silva, Carla M.F. da; Queiros, Yure G.C.; Lucas, Elizabete F. [Universidade Federal do Rio de Janeiro, Instituto de Macromoleculas, Laboratorio de Macromoleculas e Coloides na Industria de Petroleo, Rio de Janeiro, RJ (Brazil)], e-mail: elucas@ima.ufrj.br

    2011-07-01

    The aim of this study was to evaluate the performance of polymeric resins packed in a fixed bed eluted in semi-industrial scale for oil and greases removal disposed in synthetic oily water in different temperature conditions. For this work, columns packed with vinyl and acryl polymer-base were tested and their efficiency of oil removal was evaluated by fluorimetry technique in two different temperatures: 25 and 60 deg C, in a flow rate condition of 200 mL/min. The experimental results were very good: the removal efficiencies were above 98% in both cases. At 60 deg C, the system keep the efficiency for a longer time: no significant loss in the efficiency was observed after eluting 1,000 times of the column bed volume at 25 deg C and 2,000, at 60 deg C. This result characterizes a great potential of application in the industry. (author)

  18. Detection system for a gas chromatograph. [. cap alpha. -methylnaphthalene,. beta. -methylnapthalene

    Science.gov (United States)

    Hayes, J.M.; Small, G.J.

    1982-04-26

    A method and apparatus are described for the quantitative analysis of vaporizable compounds, and in particular of polycyclic aromatic hydrocarbons which may be induced to fluoresce. The sample to be analyzed is injected into a gas chromatography column and is eluted through a narrow orifice into a vacuum chamber. The free expansion of the eluted sample into the vacuum chamber creates a supersonic molecular beam in which the sample molecules are cooled to the extent that the excited vibrational and rotational levels are substantially depopulated. The cooled molecules, when induced to fluoresce by laser excitation, give greatly simplified spectra suitable for analytical purposes. The laser induced fluorimetry provides great selectivity, and the gas chromatograph provides quantitative transfer of the sample to the molecular beam. 3 figures, 2 tables.

  19. Sensing a Sensor: Identifying the Mechanosensory Function of Primary Cilia

    Directory of Open Access Journals (Sweden)

    Rahul M. Prasad

    2014-03-01

    Full Text Available Over the past decade, primary cilia have emerged as the premier means by which cells sense and transduce mechanical stimuli. Primary cilia are sensory organelles that have been shown to be vitally involved in the mechanosensation of urine in the renal nephron, bile in the hepatic biliary system, digestive fluid in the pancreatic duct, dentin in dental pulp, lacunocanalicular fluid in bone and cartilage, and blood in vasculature. The prevalence of primary cilia among mammalian cell types is matched by the tremendously varied disease states caused by both structural and functional defects in cilia. In the process of delineating the mechanisms behind these disease states, calcium fluorimetry has been widely utilized as a means of quantifying ciliary function to both fluid flow and pharmacological agents. In this review, we will discuss the approaches used in associating calcium levels to cilia function.

  20. Analytical methods applied to water pollution

    International Nuclear Information System (INIS)

    Baudin, G.

    1977-01-01

    A comparison of different methods applied to water analysis is given. The discussion is limited to the problems presented by inorganic elements, accessible to nuclear activation analysis methods. The following methods were compared: activation analysis: with gamma-ray spectrometry, atomic absorption spectrometry, fluorimetry, emission spectrometry, colorimetry or spectrophotometry, X-ray fluorescence, mass spectrometry, voltametry, polarography or other electrochemical methods, activation analysis-beta measurements. Drinking-water, irrigation waters, sea waters, industrial wastes and very pure waters are the subjects of the investigations. The comparative evaluation is made on the basis of storage of samples, in situ analysis, treatment and concentration, specificity and interference, monoelement or multielement analysis, analysis time and accuracy. The significance of the neutron analysis is shown. (T.G.)

  1. Recent advances in nuclear techniques for environmental radioactivity monitoring

    International Nuclear Information System (INIS)

    Kumar, Ajay; Tripathi, R.M.

    2016-01-01

    The environmental radioactivity monitoring was first started in the late 1950s following the global fallout from testing of nuclear weapons in the atmosphere. Nuclear analytical techniques are generally classified into two categories: destructive and non-destructive. Destructive techniques are carried out through several analytical methods such as α-spectrometry, liquid Scintillation counting system, solid state nuclear track detector, spectrophotometry, fluorimetry, atomic absorption spectrometry (AAS), inductively coupled plasma mass spectrometry (ICP-MS), inductively coupled plasma optical emission spectrometry (ICP-OES), chromatography techniques, electro-analytical techniques etc. However, nondestructive methods include gamma spectrometry, X-Ray fluorescence (XRF) spectrometry, neutron activation analysis (NAA) etc. The development of radiochemical methods and measurement techniques using alpha and gamma spectrometry have been described in brief

  2. Age-dependent radiation dose due to intake of uranium through drinking water in India

    International Nuclear Information System (INIS)

    Sahoo, S.K.; Mohapatra, S.; Chakrabarty, A.; Sumesh, C.G.; Tripathi, R.M.; Puranik, V.D.

    2009-01-01

    In the present study, an attempt has been made to estimate the content of uranium in drinking water in various states of India by laser fluorimetry. Depending upon the rate of water intake for the different age groups, the associated radiation dose was calculated. The concentration of uranium varied between 0.1 ± 0.01 and 19.6 ± 1.8 ppb which is much lower than the drinking water guideline value of 60 ppb. The total radiation dose due to ingestion of uranium through drinking water for various age groups is found to vary from 0.14 μSv/y to 48 μSv/y. (author)

  3. Kinetic analysis of pulsed laser induced phosphorescence for uranium determination

    International Nuclear Information System (INIS)

    Serdeiro, Nelida H.

    2003-01-01

    The laser induced kinetic phosphorescence allows the uranium determination in different kind of matrices, with a lower detection limit than those reached by other spectroscopic methods. It involves the uranyl ions excitation by a pulsed dye-laser source, followed by temporal resolution of the phosphorescence. This method is used for the determination of trace quantities of uranium in aqueous solution, with a suitable complexant agent, without chemical separation before the analysis. The objective of this paper is to present the results of uranium determinations in different standard samples, water, soil, filter and urine, and a comparison with other methods such as fluorimetry, alpha spectrometry and mass spectrometry. Moreover, the measurement conditions, the advantages and disadvantages, the sample preparation, the interferences and the detection limit are described. (author)

  4. The basic principles for the assessment of occupational exposure due to intakes of radionuclides (The most important indirect methods used in the Protection and Safety Dept. for internal occupational exposure monitoring)

    International Nuclear Information System (INIS)

    Kharita, M. H.; Sakhita, K.

    2009-05-01

    This study shows the basic principles for the assessment of occupational exposure due to intakes of radionuclides in order to calculate the committed effective dose of each radionuclide separately. We also discussed when the routine monitoring of workers becomes useful and when the workplace monitoring is better than workers monitoring. In addition, this study contains the details of four indirect methods as they are validated in the protection and safety department, and their names are: Determination the concentration of total uranium by using fluorimetry technique, Determination the activity of two uranium isotopes 238 and 234, The activity of Polonium 210, and the activity of Radium 226 by using alpha spectrometry for urine samples collected from workers occupationally exposed to this isotope. (author)

  5. Evaluation of polymeric materials packed in fixed bed column for oil water remediation; Avaliacao de materiais polimericos empacotados em colunas de leito fixo para a remediacao de aguas oleosas

    Energy Technology Data Exchange (ETDEWEB)

    Queiros, Yure G.C.; Barros, Cintia Chagas; Oliveira, Roberta S.; Marques, Luiz R.S.; Cunha, Luciana; Lucas, Elizabete F. [Universidade Federal do Rio de Janeiro (UFRJ), RJ (Brazil). Inst. de Macromoleculas Eloisa Mano], e-mail: yuregomes@ima.ufrj.br, e-mail: elucas@ima.ufrj.br

    2007-07-01

    Polymeric resins are being tried as an alternative material for treating oily waters from the petroleum industry, which have already been treated by conventional methods. The objective of this work has been to evaluate the purification degree of synthetic oily waters when treated in fixed bed columns packed with polymeric resins made up of hydrophilic and lipophilic moieties. The analysis used for characterizing the total grease and oil content (TOG) was fluorimetry. Starting oily waters of average TOG 50 ppm were prepared. Data obtained from eluted waters did not outweigh 10% of the TOG values of starting solutions in some blends of resins with a pretty good mechanical stability under the increase of pressure. Organoclay material showed a good retention performance, but has presented a mechanical instability too, compromising its use for larger amounts of wastewater. (author)

  6. Concentrations of polycyclic aromatic hydrocarbons in four species of bottom dwelling fish and two species of crustaceans from the Estuary and Gulf of Saint-Lawrence and from the Saguenay fjord; Concentrations d`hydrocarbures aromatiques polycycliques chez quatre especes de poissons de fond et deux especes de crustaces de l`estuaire et du golfe du Saint-Laurent et du fjord du Saguenay

    Energy Technology Data Exchange (ETDEWEB)

    Pelletier, E.; Canuel, G.; Padros, J.; Clermont, Y. [Quebec Univ., INRS-Oceanologie, Rimouski, PQ (Canada); Gobeil, C. [Fisheries and Oceans Canada, Mont-Joli, PQ (Canada). Maurice Lamontagne Inst.

    1999-08-01

    An update was presented on the levels of polycyclic aromatic hydrocarbon (PAH) contamination in bottom dwelling fish and crustaceans in the Gulf of St. Lawrence and the Saguenay fjord. Attempts were made to detect specific compounds including fluoranthene, pyrene, benzo-b-fluoranthene, benzo-k-fluoranthene, benzo-a-pyrene, dibenzo-a,h-anthracene, benzo-g,h,i-perylene, naphtalene, acenaphtene, fluorene, phenanthrene, and anthracene. Concentrations of all PAHs in the muscle tissue of cod, plaice, skate and halibut were found to be below analytical detection limits. Concentrations of most PAHs were also below detection limits in halibut liver tissue and crab. The exception was benzo-a-anthracene. Shrimp muscle contained measurable levels of many PAHs. The levels were detected by fluorimetry but could not be confirmed by mass spectroscopy. Based on these results, it was concluded that the fish can be considered safe for human consumption. 3 refs., 13 tabs., 1 fig.

  7. On the use of the spectroscopic techniques to model the interactions between radionuclides and solid minerals

    Energy Technology Data Exchange (ETDEWEB)

    Simoni, E. [IPN, Paris XI University, 91406 Orsay (France)]. e-mail: simoni@ipno.in2p3.fr

    2004-07-01

    In order to determine the radionuclides sorption constants on solid natural minerals, both thermodynamic and structural investigations, using spectroscopic techniques, are presented. The natural clays, that could be used as engineering barrier in the nuclear waste geological repository, are rather complex minerals. Therefore, in order to understand how these natural materials retain the radionuclides, it is necessary first to perform these studies on simple substrates such as phosphates, oxides and silicates (as powder and single crystal as well) and then extrapolate the obtained results on the natural minerals. As examples, the main results on the sorption processes of the hexavalent uranium onto zircon (ZrSiO{sub 4}) and lanthanum phosphate (LaPO{sub 4}) are presented. The corresponding sorption curves are simulated using the results obtained with the following spectroscopic techniques: laser induced spectro fluorimetry, X-ray photoelectron spectroscopy (XP S), X-ray absorption spectroscopy (Exafs). Finally, the thermodynamic sorption constants are calculated. (Author)

  8. Dual fluorescence of N-phenylanthranilic acid: Effect of solvents, pH and β-cyclodextrin

    Science.gov (United States)

    Rajendiran, N.; Balasubramanian, T.

    2007-11-01

    Spectral characteristics of N-phenylanthranilic acid (NPAA) have been studied in different solvents, pH and β-cyclodextrin (β-CD) and compared with anthranilic acid (2-aminobenzoic acid, 2ABA). In all solvents a dual fluorescence is observed in NPAA, whereas 2ABA gives single emission. Combining the results observed in the absorption, fluorescence emission and fluorescence excitation spectra, it is found that strong intramolecular hydrogen bonding (IHB) interactions present in NPAA molecule. The inclusion complex of NPAA with β-CD is analysed by UV-vis, fluorimetry, FT-IR, 1H NMR, scanning electron microscope and AM 1 method. The above spectral studies show that NPAA forms a 1:1 inclusion complex with β-CD and COOH group present in the β-CD cavity. A mechanism is proposed to explain the inclusion process.

  9. Dual fluorescence of N-phenylanthranilic acid: Effect of solvents, pH and beta-cyclodextrin.

    Science.gov (United States)

    Rajendiran, N; Balasubramanian, T

    2007-11-01

    Spectral characteristics of N-phenylanthranilic acid (NPAA) have been studied in different solvents, pH and beta-cyclodextrin (beta-CD) and compared with anthranilic acid (2-aminobenzoic acid, 2ABA). In all solvents a dual fluorescence is observed in NPAA, whereas 2ABA gives single emission. Combining the results observed in the absorption, fluorescence emission and fluorescence excitation spectra, it is found that strong intramolecular hydrogen bonding (IHB) interactions present in NPAA molecule. The inclusion complex of NPAA with beta-CD is analysed by UV-vis, fluorimetry, FT-IR, (1)H NMR, scanning electron microscope and AM 1 method. The above spectral studies show that NPAA forms a 1:1 inclusion complex with beta-CD and COOH group present in the beta-CD cavity. A mechanism is proposed to explain the inclusion process.

  10. Radionuclides retention: from macroscopic to microscopic

    International Nuclear Information System (INIS)

    Simoni, E.

    2002-01-01

    In order to determine the radionuclides' sorption constants on solid natural minerals, both thermodynamic and structural investigations, using spectroscopic techniques, are presented. The natural clays, that could be used as engineering barriers in the nuclear waste geological repository, are rather complex minerals. Therefore, in order to understand how these natural materials retain the radionuclides, it is necessary first to perform these studies on simple substrates such as single crystal, oxides and silicates, and then extrapolate the obtained results on the natural minerals. We examine in this paper the sorption processes of the hexavalent uranium on zircon (ZrSiO 4 ) and the trivalent curium on a natural clay (bentonite). The corresponding sorption curves are simulated using the results obtained with the following spectroscopic techniques: laser induced spectro-fluorimetry, X-ray photoelectron spectroscopy (XPS), X-ray absorption spectroscopy (EXAFS), diffuse reflectance infrared Fourier transform (DRIFT). (authors)

  11. The use of room temperature phosphorescence for the determination of uranium in tin-tailings mineral samples

    International Nuclear Information System (INIS)

    Meor Yusof bin Meor Sulaiman

    1988-01-01

    The possibility of using phosphorescence technique in determining uranium in mineral samples and its comparison with that of fluorescence using high carbonate flux is presented. Samples used are tin-tailings mineral such as monazite, xenotime, ilmenite and zircon. The calibration graph obtained shows a linear relationship between the concentration range of 0-55 ppm U. From here, analysis of the standard showed that the result obtained and that of the certified value are consistent. HN0 3 :H 2 SO 4 (1:3) and phosphoric acid leaching methods are tried and the results show that phosphoric acid is the better method for phosphate mineral. Comparison of the results obtained from this technique and that of the direct and extraction methods of fluorimetry are also made. Phosphorescence is found to be a better method in determining uranium in this type of samples. (author)

  12. Verification of the visual control efficacity on quenching effect produced by Cr, Mn, Co and Fe in the uranium fluorimetric determination, by quantifying these metals with energy dispersive X-ray fluorescence

    International Nuclear Information System (INIS)

    Morales Sanchez, E.A.

    1986-08-01

    It's described the APDC preconcentration procedure, applied to the quantification of chromium, manganese, iron and cobalt in river sediments which have been digested in order to evaluate their uranium content by fluorimetry. The purpose was to show the lack of correlation between the visual control of the quenching effect and its actual presence. A Cd-109 source and a Si-Li detector were used. Thin film method for avoiding matrix effects and a deconvolution soft-ware, AXIL, for correcting peaks overlapping were applied. Precision and accuracy were 2% and 10% respectively in the 2 to 5 micro-grams/milliliter range. It was shown that for the samples analyzed there was not a good correlation between the visual observations and the quenching effect. It was found that the principal - quencher was the iron, present in so high concentration that is caused up to 80% of attenuation. (author)

  13. Radiological Study Investigation of the Black Sand Region of the North-East of the Nile Delta

    International Nuclear Information System (INIS)

    Moafy, W.A.; El-Tahawy, M.S.

    2009-01-01

    Within a comprehensive radiological investigation of the Mediterranean cost of Egypt an environmental radiation survey of the black sand region of the North-East of Nile Delta was carried out. The activity concentration of the natural 226 Ra , 238 U, 232 Th and 40 K for 18 soil samples, 25 shore sediment samples and 6 bottom sediment samples were determined using gamma spectrometers based on HpGe detector and were found to reach 193 Bq/kg for 226 Ra and 267 Bq/kg of 232 Th for some shore sediment samples. Ten sea water samples were analyzed using laser fluorimetry technique after applying a radiochemical separation procedure and the determined total uranium concentration in these samples was found to be in the range (5.2-21.8) mBq/lt.

  14. The relationship between uranium in blood and the number of working years in the Syrian phosphate mines

    International Nuclear Information System (INIS)

    Othman, I.

    1993-01-01

    Since phosphate contains uranium, workers in phosphate mines may be expected to be exposed to radioactive elements from this source. Uranium is concentrated in three main areas in the body: bone, liver and kidney. The author chose three carriers of uranium, blood, urine and hair to study the relationship between uranium concentration and the number of working years spent in the mine. Uranium was measured in samples from workers and their families by fluorimetry. The quenching effect of blood, urine and heir on uranium standards was determined. The results show that uranium concentrations (in blood) increase with the number of working year in the mine. In addition, it has also been determined that uranium concentrations in blood samples of families living near the mine are higher than those in families living in Damascus city. Finally, it has been found that hair is not a good biological indicator for this type of study. (author). 13 refs., 4 tabs., 3 figs

  15. Laser fluorimetric determination of uranium in environmental samples from Nile Delta and adjacent regions

    International Nuclear Information System (INIS)

    Shawky, S.; Ibrahiem, N.; Farouk, A.; Ghods, A.

    1994-01-01

    Total uranium content was determined in soil and plant samples obtained from various areas in the Nile Delta. Samples taken from east and west of the delta, Suez canal cities and from the Alexandria region were analysed using laser fluorimetry (LF). Uranium was extracted from digested samples with methyl-isobutyl ketone and measured using a laser fluorimeter. The radium content of the same soil samples was determined using gamma spectroscopy. The uranium content of plant samples was determined using LF, since this technique has a detection limit lower than that of GS. Uranium content in the samples varied between 0.6-4.4 μg/g for soil and 0.032-0.17 μg/g for plant tissue. (author)

  16. Large-scale biophysical evaluation of protein PEGylation effects

    DEFF Research Database (Denmark)

    Vernet, Erik; Popa, Gina; Pozdnyakova, Irina

    2016-01-01

    PEGylation is the most widely used method to chemically modify protein biopharmaceuticals, but surprisingly limited public data is available on the biophysical effects of protein PEGylation. Here we report the first large-scale study, with site-specific mono-PEGylation of 15 different proteins...... of PEGylation on the thermal stability of a protein based on data generated by circular dichroism (CD), differential scanning calorimetry (DSC), or differential scanning fluorimetry (DSF). In addition, DSF was validated as a fast and inexpensive screening method for thermal unfolding studies of PEGylated...... proteins. Multivariate data analysis revealed clear trends in biophysical properties upon PEGylation for a subset of proteins, although no universal trends were found. Taken together, these findings are important in the consideration of biophysical methods and evaluation of second...

  17. Geochemical Survey of Pernambuco

    International Nuclear Information System (INIS)

    Horowitz, A.; Duarte, P.J.; Almeida, M.G. de; Medeiros, M.O.

    1988-01-01

    The area studied i this work is located in a triangle formed by the Sibiro and Boca da Mata Sugar-Mills and Serinhaem country. In the Cabo Formation the search determinated conglomerates, arcos and clays. Although the highest geochemical activity have been done in the decomposed crystalin, and the values from Cabo Formation don't be encourager, this formation has lithology compatible with uranium mineralization. The Cabo Formation's sediments presents lithologic variations very expressives, with conglomerates, arcoses and clay silts, which determinate the choise of the area. This area presented favorable to uranium prospecting and to others elements interesting to ragional geochemistry. The atomic absorption analysis, fluorimetry and spectrometry were done for the following elements: Zn, V, Ti, Ni, Pb, Mn, Ga, Cu, Co, Bi, Ag, B, Mo, and U. (C.D.G.) [pt

  18. Report on intercomparisons IAEA/S-17, S-18, and S-19 of the determinaton of uranium in uranium phosphate ores

    International Nuclear Information System (INIS)

    Pszonicki, L.; Hanna, A.N.; Suschny, O.

    1984-08-01

    The aim of the reported intercomparisons, organized by the IAEA's Analytical Quality Control Service, was to provide an opportunity to the participating laboratories to check the reliability of their results by comparing them with the results obtained by other laboratories. The participants were requested to determine the concentration of uranium in three samples of Brasilian uranium phosphate ores containing uranium of a low, a medium, and a large concentration. Twenty-four laboratories from 19 countries returned results. The analytical method most frequently used was radiometry of natural isotopes, followed by neutron activation analysis, fluorimetry, X-ray fluorescence and spectrophotometry. In general, the results can be considered as satisfactory. The relative confidence intervals of the medians extend in the range from -3% to +5%. The overall medians of all materials were accepted as reliable for certification

  19. Determination of uranium in samples containing bulk aluminium

    International Nuclear Information System (INIS)

    Das, S.K.; Kannan, R.; Dhami, P.S.; Tripathi, S.C.; Gandhi, P.M.

    2015-01-01

    The determination of uranium is of great importance in PUREX process and need to be analyzed at different concentration ranges depending on the stage of reprocessing. Various techniques like volumetry, spectrophotometry, ICP-OES, fluorimetry, mass spectrometry etc. are used for the measurement of uranium in these samples. Fast and sensitive methods suitable for low level detection of uranium are desirable to cater the process needs. Microgram quantities of uranium are analyzed by spectrophotometric method using 2-(5- bromo-2-pyridylazo-5-diethylaminophenol) (Br-PADAP) as the complexing agent. But, the presence of some of the metal ions viz. Al, Pu, Zr etc. interferes in its analysis. Therefore, separation of uranium from such interfering metal ions is required prior to its analysis. This paper describes the analysis of uranium in samples containing aluminium as major matrix

  20. Mineralogical and geochemical investigation of the Oligocene sedimentary series of Saint-Agoulin; Etude mineralogique et geochimique de la serie sedimentaire oligocene de Saint-Agoulin

    Energy Technology Data Exchange (ETDEWEB)

    Mohadjer, K [Commissariat a l' Energie Atomique, 92 - Fontenay-aux-Roses (France). Centre d' Etudes Nucleaires, direction des productions; Faculte des Sciences de l' Universite de Paris, 75 (France)

    1967-07-05

    The mineralogical investigation is based on determination of the amounts of quartz, clays, feldspars and on grain size analysis. A distinction was established between two lithological groups: the lower group with dominant silica the upper with dominant carbonates. Clays of the two groups are comparable with dominant illite, and accessory kaolinite and montmorillonite. The amounts of the elements Si, Al, Fe, Ca, Mg, Na, K, Ti, Mn, P, Ba, Sr, Rb, Pb, Cu, Ni, Co, V, Be, B, Zr, U, were determined by several analytical methods: chemistry, X ray fluorescence, spectrography and fluorimetry. Correlations between the chemical elements and the mineralogical components were computed and plotted. Lastly the conditions of filling up of the basin are defined, taking into account the Oligocene climatic conditions. (author) [French] L'etude mineralogique est basee sur les dosages du quartz, de la calcite, des argiles, des feldspaths et sur les granulometries. On a distingue 2 familles lithologiques: l'une riche en quartz dans la partie inferieure, l'autre riche en carbonate dans la partie superieure. Les argiles des 2 familles sont comparables: illite predominante, kaolinite et montmorillonite accessoires. Les elements: Si, Al, Fe, Ca, Mg, Na, K, Ti, Mn, P, Ba, Sr, Rb, Pb, Cu, Ni, Co, V, Be, B, Zr, U, ont ete doses par differentes methodes d'analyse: analyse chimique, fluorescence X, spectrographie et fluorimetrie. Une etude de correlation entre les elements chimiques et les constituants mineralogiques a ete realisee. Enfin, les conditions de remplissage du Bassin sont definies, en tenant compte du contexte climatique oligocene. (auteur)

  1. Fusion of microlitre water-in-oil droplets for simple, fast and green chemical assays.

    Science.gov (United States)

    Chiu, S-H; Urban, P L

    2015-08-07

    A simple format for microscale chemical assays is proposed. It does not require the use of test tubes, microchips or microtiter plates. Microlitre-range (ca. 0.7-5.0 μL) aqueous droplets are generated by a commercial micropipette in a non-polar matrix inside a Petri dish. When two droplets are pipetted nearby, they spontaneously coalesce within seconds, priming a chemical reaction. Detection of the reaction product is accomplished by colorimetry, spectrophotometry, or fluorimetry using simple light-emitting diode (LED) arrays as the sources of monochromatic light, while chemiluminescence detection of the analytes present in single droplets is conducted in the dark. A smartphone camera is used as the detector. The limits of detection obtained for the developed in-droplet assays are estimated to be: 1.4 nmol (potassium permanganate by colorimetry), 1.4 pmol (fluorescein by fluorimetry), and 580 fmol (sodium hypochlorite by chemiluminescence detection). The format has successfully been used to monitor the progress of chemical and biochemical reactions over time with sub-second resolution. A semi-quantitative analysis of ascorbic acid using Tillman's reagent is presented. A few tens of individual droplets can be scanned in parallel. Rapid switching of the LED light sources with different wavelengths enables a spectral analysis of multiple droplets. Very little solid waste is produced. The assay matrix is readily recycled, thus the volume of liquid waste produced each time is also very small (typically, 1-10 μL per analysis). Various water-immiscible translucent liquids can be used as the reaction matrix: including silicone oil, 1-octanol as well as soybean cooking oil.

  2. Semi-Automated Hydrophobic Interaction Chromatography Column Scouting Used in the Two-Step Purification of Recombinant Green Fluorescent Protein

    Science.gov (United States)

    Murphy, Patrick J. M.

    2014-01-01

    Background Hydrophobic interaction chromatography (HIC) most commonly requires experimental determination (i.e., scouting) in order to select an optimal chromatographic medium for purifying a given target protein. Neither a two-step purification of untagged green fluorescent protein (GFP) from crude bacterial lysate using sequential HIC and size exclusion chromatography (SEC), nor HIC column scouting elution profiles of GFP, have been previously reported. Methods and Results Bacterial lysate expressing recombinant GFP was sequentially adsorbed to commercially available HIC columns containing butyl, octyl, and phenyl-based HIC ligands coupled to matrices of varying bead size. The lysate was fractionated using a linear ammonium phosphate salt gradient at constant pH. Collected HIC eluate fractions containing retained GFP were then pooled and further purified using high-resolution preparative SEC. Significant differences in presumptive GFP elution profiles were observed using in-line absorption spectrophotometry (A395) and post-run fluorimetry. SDS-PAGE and western blot demonstrated that fluorometric detection was the more accurate indicator of GFP elution in both HIC and SEC purification steps. Comparison of composite HIC column scouting data indicated that a phenyl ligand coupled to a 34 µm matrix produced the highest degree of target protein capture and separation. Conclusions Conducting two-step protein purification using the preferred HIC medium followed by SEC resulted in a final, concentrated product with >98% protein purity. In-line absorbance spectrophotometry was not as precise of an indicator of GFP elution as post-run fluorimetry. These findings demonstrate the importance of utilizing a combination of detection methods when evaluating purification strategies. GFP is a well-characterized model protein, used heavily in educational settings and by researchers with limited protein purification experience, and the data and strategies presented here may aid in

  3. Report on the intercomparison run F-1 U and K(Th) in feldspar

    International Nuclear Information System (INIS)

    1990-01-01

    The present report deals with a recent intercomparison on the determination of uranium, thorium and potassium in potassium feldspar, carried out in 1976-77 and involving the participation of 20 institutes who returned a total of 196 individual results on three elements. Analytical techniques employed by the participating institutes included gamma spectrometry, emission spectroscopy, atomic absorption, X-ray fluorescence, fluorimetry and neutron activation. Each of these techniques has its own particular weakness: The basic principles of gamma-spectrometric technique for the determination of U, Th and K are well known. It is, however, difficult to determine the low concentrations of radioactive nuclides which can be found in geological materials due to systematic errors arising from non-equilibration of decay chains in the samples and from other sources including calibration. Determination of uranium in geological materials by non-destructive neutron activation analysis also has some problems. Elements such as Th, Ta, Pt, Ru, Zn and Ba when irradiated in a nuclear reactor with thermal and epithermal neutrons yield radioactive isotopes, the gamma rays of which interfere with those used in uranium determination. Detection limits for three elements U, Th and K for thermal neutron activation have been reported as 0.7, 1.0 and 10 ppb respectively, however, this seems over-optimistic to expect in real-life situations. Measurement of delayed neutrons and fission track methods for uranium and thorium also have interference problems. Fluorimetry is a suitable method for uranium at low concentrations (down to 0.1 ppm) but one has to take care to overcome quenching by removing, or at least diluting, interfering elements. The determination of some important elements in geochemical samples is often difficult because of their small concentration as well as of the presence of interfering matrix elements in the samples. Some systematic differences seem to exist between the methods

  4. Optical spectroscopy of Cm{sup 3+} in the elpasolite Cs{sub 2} NaYCl{sub 6}. Application in the structural study of Cm(III) sorption on phosphate materials; Spectroscopie optique de Cm{sup 3+} dans l`elpasolite Cs{sub 2}NaYCl{sub 6}. Application a l`etude structurale de la sorption de Cm(III) sur des materiaux phosphates

    Energy Technology Data Exchange (ETDEWEB)

    Cavellec, Ronan [Universite de Paris Sud, 91 - Orsay (France)

    1998-05-15

    Radwaste management involves important research. In order to contribute to this program we proposed the examination of two different and complementary aspects of nuclear waste: the detection by laser spectroscopy of trivalent actinide ions diluted in a crystal host and the migration of radionuclides through different barriers in geosphere that is related to understanding and modelling the sorption processes. In both cases the spectro-fluorimetry laser is used to investigate different phenomenological processes at microscopic scale. The first part of the thesis deals with the investigation of Cm{sup 3+} energy levels in Cs{sub 2}NaYCl{sub 6}. From the analysis of excitation and emission spectra 52 crystal-field levels have been assigned and the phenomenological parameters calculated. A discussion about these parameters, compared with those in case of LaCl{sub 3} and ThO{sub 2} cases, is given. An intense green fluorescence has been found for the first time for Cm{sup 3+} in elpasolite. Thus, this material seems to be a promising solid for the analytical application of actinides such as Cm{sup 3+} and Am{sup 3+} ions. In the second part the sorption of curium (5{center_dot}10{sup -8} mol {center_dot}l{sup -1}) onto phosphate materials was studied. The ionic strength, electrolyte nature and complexing agent were investigated. The laser spectro-fluorimetry is shown to be a powerful technique. The formation of surface complexes for ZrP{sub 2}O{sub 7}, Zr{sub 2}O(PO{sub 4}){sub 2} and Th{sub 4}(PO{sub 4}){sub 4}P{sub 2}O{sub 7} has been demonstrated. Fluorescence lifetime measurements indicate that in complexing medium the aqueous complex of Cm is sorbed preferentially to the Cm{sup 3+} free ion onto the solid 82 refs., 54 figs., 35 tabs.

  5. Metalloproteinases and their inhibitors are influenced by inhalative glucocorticoid therapy in combination with environmental dust reduction in equine recurrent airway obstruction.

    Science.gov (United States)

    Barton, Ann Kristin; Shety, Tarek; Bondzio, Angelika; Einspanier, Ralf; Gehlen, Heidrun

    2016-12-09

    Overexpression of matrix-metalloproteinases (MMPs) has been shown to lead to tissue damage in equine recurrent airway obstruction (RAO), as a misbalance with their natural inhibitors, the tissue inhibitors of metalloproteinases (TIMPs), occurs. This favors irreversible pulmonary fibrosis formation. Increased levels of MMPs, TIMPs or altered ratios between them can be used as biomarkers of respiratory disease. We hypothesized that levels of MMPs, TIMPs and their ratios correlate with improvement in clinical findings and bronchoalveolar lavage fluid (BALF) cytology after 10 days of inhalative glucocorticoid therapy and environmental dust reduction (EDR) and may be used to monitor treatment success. Ten horses with a history of RAO participated in a prospective clinical study. Clinical and cytological scoring was performed before and after inhalative therapy using budesonide (1500 μg BID over 10 days) and EDR (bedding of wood shavings and wet hay as roughage). Gelatin zymography was performed for qualitative and semi-quantitative evaluation of MMP-2 and MMP-9 in BALF supernatant, while fluorimetry was used to evaluate MMP-8 activity. Additionally, specific equine ELISA assays were used for quantitative assessment of MMP-2, MMP-9, TIMP-1 and TIMP-2. A significant reduction in the total and several single parameters of the clinical score were found after 10 days of inhalative therapy and EDR. The concentrations of MMP-2, MMP-9, TIMP-1 and TIMP-2 (ELISA) as well as their activities (MMP-2 and MMP-9 zymography and MMP-8 fluorimetry) were significantly decreased after therapy. Significant improvements in MMP-8/TIMP-1 and MMP-8/TIMP-2 ratios were also found, differences between other ratios before and after therapy were insignificant. Metalloproteinases and their inhibitors, in particular MMP-9 and TIMP-2, are valuable markers for clinical improvement in RAO.

  6. The Usher Syndrome Type IIIB Histidyl-tRNA Synthetase Mutation Confers Temperature Sensitivity.

    Science.gov (United States)

    Abbott, Jamie A; Guth, Ethan; Kim, Cindy; Regan, Cathy; Siu, Victoria M; Rupar, C Anthony; Demeler, Borries; Francklyn, Christopher S; Robey-Bond, Susan M

    2017-07-18

    Histidyl-tRNA synthetase (HARS) is a highly conserved translation factor that plays an essential role in protein synthesis. HARS has been implicated in the human syndromes Charcot-Marie-Tooth (CMT) Type 2W and Type IIIB Usher (USH3B). The USH3B mutation, which encodes a Y454S substitution in HARS, is inherited in an autosomal recessive fashion and associated with childhood deafness, blindness, and episodic hallucinations during acute illness. The biochemical basis of the pathophysiologies linked to USH3B is currently unknown. Here, we present a detailed functional comparison of wild-type (WT) and Y454S HARS enzymes. Kinetic parameters for enzymes and canonical substrates were determined using both steady state and rapid kinetics. Enzyme stability was examined using differential scanning fluorimetry. Finally, enzyme functionality in a primary cell culture was assessed. Our results demonstrate that the Y454S substitution leaves HARS amino acid activation, aminoacylation, and tRNA His binding functions largely intact compared with those of WT HARS, and the mutant enzyme dimerizes like the wild type does. Interestingly, during our investigation, it was revealed that the kinetics of amino acid activation differs from that of the previously characterized bacterial HisRS. Despite the similar kinetics, differential scanning fluorimetry revealed that Y454S is less thermally stable than WT HARS, and cells from Y454S patients grown at elevated temperatures demonstrate diminished levels of protein synthesis compared to those of WT cells. The thermal sensitivity associated with the Y454S mutation represents a biochemical basis for understanding USH3B.

  7. DNA degrades during storage in formalin-fixed and paraffin-embedded tissue blocks.

    Science.gov (United States)

    Guyard, Alice; Boyez, Alice; Pujals, Anaïs; Robe, Cyrielle; Tran Van Nhieu, Jeanne; Allory, Yves; Moroch, Julien; Georges, Odette; Fournet, Jean-Christophe; Zafrani, Elie-Serge; Leroy, Karen

    2017-10-01

    Formalin-fixed paraffin-embedded (FFPE) tissue blocks are widely used to identify clinically actionable molecular alterations or perform retrospective molecular studies. Our goal was to quantify degradation of DNA occurring during mid to long-term storage of samples in usual conditions. We selected 46 FFPE samples of surgically resected carcinomas of lung, colon, and urothelial tract, of which DNA had been previously extracted. We performed a second DNA extraction on the same blocks under identical conditions after a median period of storage of 5.5 years. Quantitation of DNA by fluorimetry showed a 53% decrease in DNA quantity after storage. Quantitative PCR (qPCR) targeting KRAS exon 2 showed delayed amplification of DNA extracted after storage in all samples but one. The qPCR/fluorimetry quantification ratio decreased from 56 to 15% after storage (p DNA analyzable by qPCR represented only 11% of the amount obtained at first extraction. Maximal length of amplifiable DNA fragments assessed with a multiplex PCR was reduced in DNA extracted from stored tissue, indicating that DNA fragmentation had increased in the paraffin blocks during storage. Next-generation sequencing was performed on 12 samples and showed a mean 3.3-fold decrease in library yield and a mean 4.5-fold increase in the number of single-nucleotide variants detected after storage. In conclusion, we observed significant degradation of DNA extracted from the same FFPE block after 4 to 6 years of storage. Better preservation strategies should be considered for storage of FFPE biopsy specimens.

  8. Translocation of iron from lysosomes to mitochondria during acetaminophen-induced hepatocellular injury: Protection by starch-desferal and minocycline.

    Science.gov (United States)

    Hu, Jiangting; Kholmukhamedov, Andaleb; Lindsey, Christopher C; Beeson, Craig C; Jaeschke, Hartmut; Lemasters, John J

    2016-08-01

    Acetaminophen (APAP) overdose causes hepatotoxicity involving mitochondrial dysfunction and the mitochondrial permeability transition (MPT). Iron is a critical catalyst for ROS formation, and reactive oxygen species (ROS) play an important role in APAP-induced hepatotoxicity. Previous studies show that APAP disrupts lysosomes, which release ferrous iron (Fe(2+)) into the cytosol to trigger the MPT and cell killing. Here, our aim was to investigate whether iron released from lysosomes after APAP is then taken up into mitochondria via the mitochondrial electrogenic Ca(2+), Fe(2+) uniporter (MCFU) to cause mitochondrial dysfunction and cell death. Hepatocytes were isolated from fasted male C57BL/6 mice. Necrotic cell killing was assessed by propidium iodide fluorimetry. Mitochondrial membrane potential (ΔΨ) was visualized by confocal microscopy of rhodamine 123 (Rh123) and tetramethylrhodamine methylester (TMRM). Chelatable Fe(2+) was monitored by quenching of calcein (cytosol) and mitoferrofluor (MFF, mitochondria). ROS generation was monitored by confocal microscopy of MitoSox Red and plate reader fluorimetry of chloromethyldihydrodichlorofluorescein diacetate (cmH2DCF-DA). Administered 1h before APAP (10mM), the lysosomally targeted iron chelator, starch-desferal (1mM), and the MCFU inhibitors, Ru360 (100nM) and minocycline (4µM), decreased cell killing from 83% to 41%, 57% and 53%, respectively, after 10h. Progressive quenching of calcein and MFF began after ~4h, signifying increased cytosolic and mitochondrial chelatable Fe(2+). Mitochondria then depolarized after ~10h. Dipyridyl, a membrane-permeable iron chelator, dequenched calcein and MFF fluorescence after APAP. Starch-desferal, but not Ru360 and minocycline, suppressed cytosolic calcein quenching, whereas starch-desferal, Ru360 and minocycline all suppressed mitochondrial MFF quenching and mitochondrial depolarization. Starch-desferal, Ru360 and minocycline also each decreased ROS formation. Moreover

  9. Interactions between radionuclides and organic colloids. Structure and reactivity of humic compounds

    International Nuclear Information System (INIS)

    Plancque, G.

    2001-09-01

    Humic compounds are the main organic colloids present in natural waters. These compounds can significantly modify the speciation of metals and control their properties, like migration, toxicity or bio-availability. It is thus important to study their speciation in conditions representative to those encountered in the natural environment. The aim of this work is to analyze the reactivity of these humic compounds. Two spectroscopic techniques have been used: the time-resolution laser spectro-fluorimetry, limited to the study of fluorescent elements, and the electro-spray source mass spectroscopy which requires the development of specific protocols for all elements of the periodic classification system. Europium, a fluorescent element analogue to trivalent actinides, has been chosen as test-metal for the intercomparison of both spectroscopic techniques. The first technique has permitted to determine the inorganic and organic speciation (spectra and lifetime of europium hydroxides and carbonates, and constants of interaction with humic acids, respectively). The limitations of this technique in the study of inorganic speciation has been evidenced. Humic compounds have a badly defined structure. The use of high-resolution mass spectroscopy has permitted to propose in a direct and experimental way, a molecular structure of aquatic fulvic acids in agreement with their known physico-chemical properties. (J.S.)

  10. Results of the analysis of the intercomparison samples of the depleted uranium dioxide SR-20

    International Nuclear Information System (INIS)

    Aigner, H.; Deron, S.; Kuhn, E.; Ronesch, K.; Zoigner, A.

    Samples of a homogeneous powder of depleted uranium dioxide, SR-20, were distributed to 32 laboratories in January 1980 for intercomparison of the precisions and accuracies of wet chemical assay. 11 laboratories reported their results (ANNEX 1). 5 laboratories applied titration procedures, 4 of them applied methods derived from the Davies and Gray procedure (1), 2 laboratories used controlled potential coulometry, 2 laboratories used precipitation procedures, 1 laboratory used fluorimetry and 1 laboratory used activation analysis. An analysis of variance yields for each laboratory the estimates of the measurement errors, the dissolution or treatment errors and the random calibration errors. The measurement errors vary between 0.01% and 1.7% relative. The differences to the reference value vary between -9.1% and +0.92% uranium, but 9 laboratories agree within +-1%U with the reference value. The mean bias of these 9 laboratories is equal to +0.04%U. The standard deviation of the biases of these 9 laboratories is equal to 0.36%.U

  11. Spectral pathways for exploration of secondary uranium: An investigation in the desertic tracts of Rajasthan and Gujarat, India

    Science.gov (United States)

    Bharti, Rishikesh; Kalimuthu, R.; Ramakrishnan, D.

    2015-10-01

    This study aims at identifying potential zones of secondary uranium enrichment using hyperspectral remote sensing, γ-ray spectrometry, fluorimetry and geochemical techniques in the western Rajasthan and northern Gujarat, India. The investigated area has suitable source rocks, conducive past-, and present-climate that can facilitate such enrichment. This enrichment process involves extensive weathering of uranium bearing source rocks, leaching of uranyl compounds in groundwater, and their precipitation in chemical deltas along with duricrusts like calcretes and gypcretes. Spatial distribution of groundwater calcretes (that are rich in Mg-calcite) and gypcretes (that are rich in gypsum) along palaeochannels and chemical deltas were mapped using hyperspectral remote sensing data based on spectral absorptions in 1.70 μm, 2.16 μm, 2.21 μm, 2.33 μm, 2.44 μm wavelength regions. Subsequently based on field radiometric survey, zones of U anomalies were identified and samples of duricrusts and groundwater were collected for geochemical analyses. Anomalous concentration of U (2345.7 Bq/kg) and Th (142.3 Bq/kg) are observed in both duricrusts and groundwater (U-1791 μg/l, Th-34 μg/l) within the palaeo-delta and river confluence. The estimated carnotite Solubility Index also indicates the secondary enrichment of U and the likelihood of occurrence of an unconventional deposit.

  12. Engineering Tocopherol Selectivity in α-TTP: A Combined In Vitro/In Silico Study

    Science.gov (United States)

    Helbling, Rachel E.; Aeschimann, Walter; Simona, Fabio; Stocker, Achim; Cascella, Michele

    2012-01-01

    We present a combined in vitro/in silico study to determine the molecular origin of the selectivity of -tocopherol transfer protein (-TTP) towards -tocopherol. Molecular dynamics simulations combined to free energy perturbation calculations predict a binding free energy for -tocopherol to -TTP 8.262.13 kcal mol lower than that of -tocopherol. Our calculations show that -tocopherol binds to -TTP in a significantly distorted geometry as compared to that of the natural ligand. Variations in the hydration of the binding pocket and in the protein structure are found as well. We propose a mutation, A156L, which significantly modifies the selectivity properties of -TTP towards the two tocopherols. In particular, our simulations predict that A156L binds preferentially to -tocopherol, with striking structural similarities to the wild-type--tocopherol complex. The affinity properties are confirmed by differential scanning fluorimetry as well as in vitro competitive binding assays. Our data indicate that residue A156 is at a critical position for determination of the selectivity of -TTP. The engineering of TTP mutants with modulating binding properties can have potential impact at industrial level for easier purification of single tocopherols from vitamin E mixtures coming from natural oils or synthetic processes. Moreover, the identification of a -tocopherol selective TTP offers the possibility to challenge the hypotheses for the evolutionary development of a mechanism for -tocopherol selection in omnivorous animals. PMID:23152872

  13. Towards an interpretation of the mechanism of the actinides(III)/lanthanides(III) separation by synergistic solvent extraction with nitrogen-containing polydendate ligands; Vers une interpretation des mecanismes de la separation actinides(III)/lanthanides(III) par extraction liquide-liquide synergique impliquant des ligands polyazotes

    Energy Technology Data Exchange (ETDEWEB)

    Francois, N [CEA/VALRHO - site de Marcoule, Dept. de Recherche en Retraitement et en Vitrification, (DRRV), 30 - Marcoule (France); Universite Henri Poincare, 54 - Vandoeuvre-les-Nancy (France)

    2000-07-01

    In the field of the separation of long-lived radionuclides from the wastes produced by nuclear fuel reprocessing, aromatic nitrogen-containing polydendate ligands are potential candidates for the selective extraction, alone or in synergistic mixture with acidic extractants, of trivalent actinides from trivalent lanthanides. The first part of this work deals with the complexation of trivalent f cations with various nitrogen-containing ligands (poly-pyridine analogues). Time-resolved laser-induced fluorimetry (TRLIF) and UV-visible spectrophotometry were used to determine the nature and evaluate the stability of each complex. Among the ligands studied, the least basic Me-Btp proved to be highly selective towards americium(III) in acidic solution. In the second part, two synergistic systems (nitrogen-containing polydendate ligand and lipophilic carboxylic acid) are studied and compared in regard to the extraction and separation of lanthanides(III) and actinides(III). TRLIF and gamma spectrometry allowed the nature of the extracted complexes and the optimal conditions of efficiency of both systems to be determined. Comparison between these different studies showed that the selectivity of complexation of trivalent f cations by a given nitrogen-containing polydendate ligand could not always be linked to the Am(III)Eu(III) selectivity reached in synergistic extraction. The latter depends on the 'balance' between the acid-basic properties on the one hand, and on the hard-soft characteristics on the other hand, of both components of synergistic system. (author)

  14. Towards an interpretation of the mechanism of the actinides(III)/lanthanides(III) separation by synergistic solvent extraction with nitrogen-containing polydendate ligands

    International Nuclear Information System (INIS)

    Francois, N.

    2000-01-01

    In the field of the separation of long-lived radionuclides from the wastes produced by nuclear fuel reprocessing, aromatic nitrogen-containing polydendate ligands are potential candidates for the selective extraction, alone or in synergistic mixture with acidic extractants, of trivalent actinides from trivalent lanthanides. The first part of this work deals with the complexation of trivalent f cations with various nitrogen-containing ligands (poly-pyridine analogues). Time-resolved laser-induced fluorimetry (TRLIF) and UV-visible spectrophotometry were used to determine the nature and evaluate the stability of each complex. Among the ligands studied, the least basic Me-Btp proved to be highly selective towards americium(III) in acidic solution. In the second part, two synergistic systems (nitrogen-containing polydendate ligand and lipophilic carboxylic acid) are studied and compared in regard to the extraction and separation of lanthanides(III) and actinides(III). TRLIF and gamma spectrometry allowed the nature of the extracted complexes and the optimal conditions of efficiency of both systems to be determined. Comparison between these different studies showed that the selectivity of complexation of trivalent f cations by a given nitrogen-containing polydendate ligand could not always be linked to the Am(III)Eu(III) selectivity reached in synergistic extraction. The latter depends on the 'balance' between the acid-basic properties on the one hand, and on the hard-soft characteristics on the other hand, of both components of synergistic system. (author)

  15. Drift curves from spray applications on commom bean crop

    Directory of Open Access Journals (Sweden)

    Mariana Rodrigues Bueno

    Full Text Available ABSTRACT In order to avoid the occurrence of drift in pesticide applications, it is fundamental to know the behavior of sprayed droplets. This study aimed to determine drift curves in pesticide applications on common bean crop under brazilian weather conditions, using different nozzle types and compared them with the "German" and "Dutch" drift prediction models. The experiment was conducted in Uberlândia, Minas Gerais/Brazil, in completely randomized design with ten replications and 4 x 20 split-plot arrangement in space. Drift deposited on collectors located over ground level was resulted by 150 L ha-1 carrier volume applications through four nozzle types (XR 11002 (fine droplets; AIXR 11002 (coarse droplets; TT 11002 (medium droplets; TTI 11002 (extremely coarse droplets, collected in 20 downwind distances, parallel to the crop line outside the target area, spaced by 2.5 m. The tracer rhodamine B was added to the spray to be quantified by fluorimetry. Drift prediction models adjusted by exponential functions were obtained considering the 90th percentile for XR, TT, AIXR and TTI nozzles. It is suggested to use the estimated drift models from this study for each nozzle type in drift prediction evaluations on bean crops under brazilian weather conditions.

  16. Study of rare earth elements, uranium and thorium migration in rocks from Espinharas uranium deposit, Paraiba - Brazil

    International Nuclear Information System (INIS)

    Conceicao, Cirilo C.S.

    2009-01-01

    The determination of rare earth elements as natural analogue in patterns geologic has grown as a tool for predicting the long-term safety of nuclear disposal in geological formation. Migration of natural radionuclides is one of the most serious problems in the waste deposit from nuclear fuel cycle. Rare earth elements show the same kinetic behavior in rocks as natural radionuclides. This similar property of the analogues allows perform studies and models on the subject of radionuclides migration. The aim of this study was to determine the distribution of rare earth elements in rocks located at Espinharas - Paraiba - Brazil, uranium deposit. In this work are presented the results from the study above the distribution of rare earth elements in function of the degree of mineralized rocks, composition and the conditions of radioactive equilibrium of the uranium and thorium in some fractures on the rocks from radioactive occurrence of Espinharas-Brazil. The results show that there is a correlation of heavy rare earth elements, uranium and Thorium concentrations to oxidation factor of the rocks. However this correlation was not observed for light rare earth elements. It means that heavy rare earth elements follow the natural radionuclides in oxidation process of rocks. The samples were analyzed by ICP-MS, alpha and gamma spectrometry, X-ray diffraction and fluorimetry. (author)

  17. The determination of boron in aluminium of high purity

    International Nuclear Information System (INIS)

    Cook, E.B.T.; Holan, H.

    1977-01-01

    A description is given of the investigations that led to the development of chemical methods for the determination of boron within the range 0,25 to 1,0 p.p.m. in aluminium of high purity. Methods were developed that incorporated fluorimetry, directly in solutions containing aluminium and after separation of boron by liquid-liquid extraction into 2-ethyl-1,3 hexanediol. A published spectrophotometric method, involving extraction of the BF 4 sup(-) complex with methylene blue into dichloroethane, was modified for application to alluminium samples. Details of this modified procedure and the fluorimetric-extraction procedure are appended. The precision of the methylene-blue method is about 6 percent relative and is recommended for precision and speed in preference to others. Separation of boron by distillation and spectrophotometric determination with curcumin gave low values in comparison with those obtained by the other methods. Agreement between the boron values obtained on the samples tested was good for the fluorimetric and methylene-blue spectrophotometric methods

  18. Submicron polymer particles containing fluorescent semiconductor nanocrystals CdSe/ZnS for bioassays.

    Science.gov (United States)

    Generalova, Alla N; Sizova, Svetlana V; Zdobnova, Tatiana A; Zarifullina, Margarita M; Artemyev, Michail V; Baranov, Alexander V; Oleinikov, Vladimir A; Zubov, Vitaly P; Deyev, Sergey M

    2011-02-01

    This study aimed to design a panel of uniform particulate biochemical reagents and to test them in specific bioassays. These reagents are polymer particles of different sizes doped with semiconductor nanocrystals and conjugated with either full-size antibodies or recombinant mini-antibodies (4D5 scFv fragment) designed by genetic engineering approaches. A panel of highly fluorescent polymer particles (150-800 nm) were formed by embedding CdSe/ZnS nanocrystals (quantum dots) into preformed polyacrolein and poly(acrolein-co-styrene) particles. Morphology, content and fluorescence characteristics of the prepared materials were studied by laser correlation spectroscopy, spectrophotometry, optical and fluorescent microscopy and fluorimetry. The obtained fluorescent particles sensitized by anti-Yersinia pestis antibodies were used for rapid agglutination glass test suitable for screening analysis of Y. pestis antigen and for microtiter particle agglutination, which, owing to its speed and simplicity, is very beneficial for diagnostic detection of Y. pestis antigen. Recombinant 4D5 scFv antibodies designed and conjugated with polymer particles containing quantum dots provide multipoint highly specific binding with cancer marker HER2/neu on the surface of SKOV-3 cell.

  19. Identification of potential glutaminyl cyclase inhibitors from lead-like libraries by in silico and in vitro fragment-based screening.

    Science.gov (United States)

    Szaszkó, Mária; Hajdú, István; Flachner, Beáta; Dobi, Krisztina; Magyar, Csaba; Simon, István; Lőrincz, Zsolt; Kapui, Zoltán; Pázmány, Tamás; Cseh, Sándor; Dormán, György

    2017-02-01

    A glutaminyl cyclase (QC) fragment library was in silico selected by disconnection of the structure of known QC inhibitors and by lead-like 2D virtual screening of the same set. The resulting fragment library (204 compounds) was acquired from commercial suppliers and pre-screened by differential scanning fluorimetry followed by functional in vitro assays. In this way, 10 fragment hits were identified ([Formula: see text]5 % hit rate, best inhibitory activity: 16 [Formula: see text]). The in vitro hits were then docked to the active site of QC, and the best scoring compounds were analyzed for binding interactions. Two fragments bound to different regions in a complementary manner, and thus, linking those fragments offered a rational strategy to generate novel QC inhibitors. Based on the structure of the virtual linked fragment, a 77-membered QC target focused library was selected from vendor databases and docked to the active site of QC. A PubChem search confirmed that the best scoring analogues are novel, potential QC inhibitors.

  20. Trypanosoma cruzi Epimastigotes Are Able to Store and Mobilize High Amounts of Cholesterol in Reservosome Lipid Inclusions

    Science.gov (United States)

    Pereira, Miria G.; Nakayasu, Ernesto S.; Sant'Anna, Celso; De Cicco, Nuccia N. T.; Atella, Georgia C.; de Souza, Wanderley; Almeida, Igor C.; Cunha-e-Silva, Narcisa

    2011-01-01

    Background Reservosomes are lysosome-related organelles found in Trypanosoma cruzi epimastigotes. They represent the last step in epimastigote endocytic route, accumulating a set of proteins and enzymes related to protein digestion and lipid metabolism. The reservosome matrix contains planar membranes, vesicles and lipid inclusions. Some of the latter may assume rectangular or sword-shaped crystalloid forms surrounded by a phospholipid monolayer, resembling the cholesterol crystals in foam cells. Methodology/Principal Findings Using Nile Red fluorimetry and fluorescence microscopy, as well as electron microscopy, we have established a direct correlation between serum concentration in culture medium and the presence of crystalloid lipid inclusions. Starting from a reservosome purified fraction, we have developed a fractionation protocol to isolate lipid inclusions. Gas-chromatography mass-spectrometry (GC-MS) analysis revealed that lipid inclusions are composed mainly by cholesterol and cholesterol esters. Moreover, when the parasites with crystalloid lipid-loaded reservosomes were maintained in serum free medium for 48 hours the inclusions disappeared almost completely, including the sword shaped ones. Conclusions/Significance Taken together, our results suggest that epimastigote forms of T. cruzi store high amounts of neutral lipids from extracellular medium, mostly cholesterol or cholesterol esters inside reservosomes. Interestingly, the parasites are able to disassemble the reservosome cholesterol crystalloid inclusions when submitted to serum starvation. PMID:21818313

  1. Light-Regulated Release of Entrapped Drugs from Photoresponsive Gold Nanoparticles

    Directory of Open Access Journals (Sweden)

    Kaniknun Sreejivungsa

    2016-01-01

    Full Text Available Release of a payload in a spatiotemporal fashion has a substantial impact on increasing therapeutic efficacy. In this work, a novel monolayer of gold nanoparticles (AuNPs featuring light-responsive ligands was investigated as a potential drug carrier whose drug release can be triggered by UV light. Hydrophobic molecules were noncovalently entrapped in the compartments of its monolayers. Once irradiated with UV light, the dinitrobenzyl linker was cleaved, leading to release of the entrapped agent. AuNPs were characterized using UV spectrophotometry, TEM, and a zetasizer. A naturally occurring compound extracted from Goniothalamus elegans Ast was chosen as a hydrophobic model drug. Entrapment and release of dye were monitored using fluorimetry. The percent encapsulation of dye was of 13.53%. Entrapped dye can be released upon UV irradiation and can be regulated by changing irradiation time. Up to 83.95±2.2% entrapped dye can be released after irradiation for 20 minutes. In the absence of UV light, dye release was only 19.75%. For comparison purposes, AuNPs having no dinitrobenzyl groups showed a minimal release of 12.23% and 11.69% with and without UV light, respectively. This demonstrated an alternative strategy to encapsulate drugs using a noncovalent approach followed by their controlled release upon UV irradiation.

  2. Water solubilization capacity of pharmaceutical microemulsions based on Peceol®, lecithin and ethanol.

    Science.gov (United States)

    Mouri, Abdelkader; Diat, Olivier; Lerner, Dan Alain; El Ghzaoui, Abdeslam; Ajovalasit, Alessia; Dorandeu, Christophe; Maurel, Jean-Claude; Devoisselle, Jean-Marie; Legrand, Philippe

    2014-11-20

    Biocompatible microemulsions composed of Peceol(®), lecithin, ethanol and water developed for encapsulation of hydrophilic drugs were investigated. The binary mixture Peceol(®)/ethanol was studied first. It was shown that the addition of ethanol to pure Peceol(®) has a significant fluidifying and disordering effect on the Peceol(®) supramolecular structure with an enhancement in water solubilization. The water solubilization capacity was improved by adding lecithin as a third component. It was then demonstrated that the ethanol/lecithin weight ratio played an important role in determining the optimal composition in term of water solubilization efficiency, a necessary property for a nutraceutical or pharmaceutical application. The optimal ethanol/lecithin weight ratio in the Peceol(®) rich region was found to be 40/60. Combination different techniques such as SAXS, fluorimetry, rheology and conductivity, we analyzed the water uptake within the microemulsion taking into account the partitioning of ethanol between polar and apolar domains. This ethanol distribution quantified along a water dilution line has a major effect on microemulsion properties. Copyright © 2014 Elsevier B.V. All rights reserved.

  3. Depolarization of the conductance-voltage relationship in the NaV1.5 mutant, E1784K, is due to altered fast inactivation

    Science.gov (United States)

    Yu, Alec; Zhu, Wandi; Silva, Jonathan R.; Ruben, Peter C.

    2017-01-01

    E1784K is the most common mixed long QT syndrome/Brugada syndrome mutant in the cardiac voltage-gated sodium channel NaV1.5. E1784K shifts the midpoint of the channel conductance-voltage relationship to more depolarized membrane potentials and accelerates the rate of channel fast inactivation. The depolarizing shift in the midpoint of the conductance curve in E1784K is exacerbated by low extracellular pH. We tested whether the E1784K mutant shifts the channel conductance curve to more depolarized membrane potentials by affecting the channel voltage-sensors. We measured ionic currents and gating currents at pH 7.4 and pH 6.0 in Xenopus laevis oocytes. Contrary to our expectation, the movement of gating charges is shifted to more hyperpolarized membrane potentials by E1784K. Voltage-clamp fluorimetry experiments show that this gating charge shift is due to the movement of the DIVS4 voltage-sensor being shifted to more hyperpolarized membrane potentials. Using a model and experiments on fast inactivation-deficient channels, we show that changes to the rate and voltage-dependence of fast inactivation are sufficient to shift the conductance curve in E1784K. Our results localize the effects of E1784K to DIVS4, and provide novel insight into the role of the DIV-VSD in regulating the voltage-dependencies of activation and fast inactivation. PMID:28898267

  4. Monitoring radioactive contamination in food, water sediments and other environmental samples in Egypt. Final report for the period 1 October 1992 - 31 July 1996

    International Nuclear Information System (INIS)

    Farouk, M.

    1996-09-01

    A detailed survey for radioactivity content was carried out in the following materials: 1) Raw (from quarries) and finished (e.g., bricks, cement, ceramic tiles) building materials originating in central Egypt; 2) soils and plants (particularly cattle fodder) in upper Egypt, including soil to plant transfer factors; 3) water, bottom sediments and shore sediments from Lake Nasser and its immediate surroundings; 4) water, fish, bottom and shore sediments from the Lake Quaran region; 5) soil, plant, drinking water, bottom and shore sediments from the Sinai Peninsula. The concentrations of the following radionuclides were measured: K-40, Cs-137, Ra-226, Th-232 and U-238. Samples were sealed in Marinelli beakers for 4 weeks to allow complete ingrowth of Rn-222 and daughters. High resolution gamma-ray spectrometry was used to measure K-40 and Cs-137 directly and Ra-226, Th-232 and U-238 indirectly by means of daughter radioactivities. In addition, uranium measurements were also made after chemical decomposition of selected samples by laser fluorimetry and by alpha spectrometry techniques. The results were used for establishing a database of baseline radioactivity levels, for evaluating average doses to the population, for comparison with similar materials from other countries and for determining soil to plant transfer factors which can be used to estimate internal doses to the population. (author). Refs, figs, tabs

  5. Structure of the effector-binding domain of the arabinose repressor AraR from Bacillus subtilis

    Energy Technology Data Exchange (ETDEWEB)

    Procházková, Kateřina; Čermáková, Kateřina [Academy of Sciences of the Czech Republic, Flemingovo nam. 2, Prague 6 (Czech Republic); Pachl, Petr; Sieglová, Irena [Academy of Sciences of the Czech Republic, Flemingovo nam. 2, Prague 6 (Czech Republic); Academy of Sciences of the Czech Republic, Videnska 1083, Prague 4 (Czech Republic); Fábry, Milan [Academy of Sciences of the Czech Republic, Videnska 1083, Prague 4 (Czech Republic); Otwinowski, Zbyszek [UT Southwestern Medical Center, Dallas, Texas (United States); Řezáčová, Pavlína, E-mail: rezacova@uochb.cas.cz [Academy of Sciences of the Czech Republic, Flemingovo nam. 2, Prague 6 (Czech Republic); Academy of Sciences of the Czech Republic, Videnska 1083, Prague 4 (Czech Republic)

    2012-02-01

    The crystal structure of the effector-binding domain of the transcriptional repressor AraR from B. subtilis in complex with the effector molecule (l-arabinose) was determined at 2.2 Å resolution. A detailed analysis of the crystal identified a dimer organization that is distinctive from that of other members of the GalR/LacI family. In Bacillus subtilis, the arabinose repressor AraR negatively controls the expression of genes in the metabolic pathway of arabinose-containing polysaccharides. The protein is composed of two domains of different phylogenetic origin and function: an N-terminal DNA-binding domain belonging to the GntR family and a C-terminal effector-binding domain that shows similarity to members of the GalR/LacI family. The crystal structure of the C-terminal effector-binding domain of AraR in complex with the effector l-arabinose has been determined at 2.2 Å resolution. The l-arabinose binding affinity was characterized by isothermal titration calorimetry and differential scanning fluorimetry; the K{sub d} value was 8.4 ± 0.4 µM. The effect of l-arabinose on the protein oligomeric state was investigated in solution and detailed analysis of the crystal identified a dimer organization which is distinctive from that of other members of the GalR/LacI family.

  6. Coordination between antioxidant defences might be partially modulated by magnesium status.

    Science.gov (United States)

    Monteiro, Cristina Paula; Matias, Catarina Nunes; Bicho, Manuel; Santa-Clara, Helena; Laires, Maria José

    2016-04-01

    The aim of this study was to compare the redox balance in competitive adult swimmers against recreational practitioners, controlling for Mg intake. Fifteen, competitive swimmers and 16 recreational practitioners, all male and aged 18-25years, were recruited into the study. Oxidative and muscle damage markers, and antioxidant enzymes and non-enzymatic antioxidants were evaluated by photometry (except for thiobarbituric reactive substances (TBARS), which was assessed by fluorimetry). Controlling for the level of exercise, inverse correlations were observed for uric acid and glutathione reductase (GR) or susceptibility of red blood cells to peroxidation (RBCPx); plasma adrenaline oxidase activity (AdOx) and carotenoids; TBARS and GR or Vit E; and direct correlations were observed between AdOx and creatine kinase (CK) or TBARS; CK and superoxide dismutase activity; GR and RBCPx. Controlling for Mg intake in addition to exercise level revealed new inverse correlations: between carotenoids and TBARS or lactate, and new direct correlations between lactate and AdOx or TBARS; cortisol and AdOx, CK, lactate dehydrogenase, or methemoglobin reductase. The associations between uric acid and RBCPx; AdOx and CK or TBARS; and GR and RBCPx lost their significance. All others remained significant. These outcomes suggest that the coordination between antioxidant defences may be partially modulated by Mg, which may be the result of its ability to stabilize cell membranes and oxidation targets, such as adrenaline.

  7. National intercomparison programme for radionuclide analysis in environmental samples: Aramar radioecological laboratory performance

    Energy Technology Data Exchange (ETDEWEB)

    Arine, Bruno Burini Robles, E-mail: bruno.arine@ctmsp.mar.mil.b [Centro Tecnologico da Marinha em Sao Paulo (CTMSP/ARAMAR), Ipero, SP (Brazil). Lab. Radioecologico; Moraes, Marco Antonio P.V., E-mail: marco.proenca@ctmsp.mar.mil.b [Instituto de Pesquisas Energeticas e Nucleares (IPEN/CNEN-SP), Sao Paulo, SP (Brazil)

    2011-07-01

    The radioecological laboratory is concerned with the measurements of background radiation (mainly uranium and thorium natural series) and present effluents in the Aramar Experimental Centre, as well as in its surroundings. The laboratory is directly subordinated to the Navy Technological Centre in Sao Paulo (CTMSP - Sao Paulo - Brazil), a military research organization whose goal is to develop nuclear and energy systems for the Brazilian naval ship propulsion. The measurements were performed in addition to the Environmental Monitoring Programme carried out in the same region. For this endeavour, the laboratory has attended to the National Intercomparison Programme conducted by the Institute for Radioprotection and Dosimetry (IRD) by analyzing several kinds of solid and liquid samples containing specific radionuclides through gamma spectrometry, liquid scintillation, alpha-beta total counting and fluorimetry techniques, since December 1995. In the last 15 years, our results were compared to another 19 laboratories and rated as 'very good' and 'acceptable' in at least 90% of the results. (author)

  8. Use of polymeric resins for removing contaminants from oily waters

    Energy Technology Data Exchange (ETDEWEB)

    Clarisse, M.D.; Queiros, Y.G.C.; Mauro, A.C.; Lucas, E.F. [Universidade Federal do Rio de Janeiro (UFRJ), RJ (Brazil). Inst. de Macromoleculas; Barbosa, C.C.R. [Instituto de Engenharia Nuclear (IEN/CNEN-RJ), Rio de Janeiro, RJ (Brazil); Barbosa, L.C.F.; Louvisse, A.M.T. [PETROBRAS, Rio de Janeiro, RJ (Brazil). Centro de Pesquisas (CENPES)

    2004-07-01

    Polymeric resins are being tried as an alternative material for treating oily waters from the petroleum industry, which have already been treated by conventional methods. The objective of this work has been to evaluate the purification degree of synthetic oily waters when treated in fixed bed columns packed with polymeric resins made up of hydrophilic and lipophilic moieties. The analysis used for characterizing the total grease and oil content (TOG) was fluorimetry. Starting oily waters of average TOG 40 ppm were prepared. Data obtained from eluted waters did not outweigh 1% of the TOG values of starting solutions. The kinetic study showed that the contaminant removal efficiency depends on the system elution flow rate; optimum removal values were reached at a 7.0 mL/min flow rate. High efficiency and speed in the purification process were obtained at this optimum flow rate. The passage of a water volume 1,000 times the volume of the column bed was not sufficient to observe its saturation level. (author)

  9. A highly active endo-levanase BT1760 of a dominant mammalian gut commensal Bacteroides thetaiotaomicron cleaves not only various bacterial levans, but also levan of timothy grass

    DEFF Research Database (Denmark)

    Mardo, Karin; Visnapuu, Triinu; Vija, Heiki

    2017-01-01

    of Pseudomonas syringae pv. tomato, its mutant Asp300Asn, levansucrases of Zymomonas mobilis, Erwinia herbicola, Halomonas smyrnensis as well as on levan isolated from timothy grass. For the first time a plant levan is shown as a perfect substrate for an endo-fructanase of a human gut bacterium. BT1760 degraded...... levans to FOS with degree of polymerization from 2 to 13. At optimal reaction conditions up to 1 g of FOS were produced per 1 mg of BT1760 protein. Low molecular weight (grass levan and levan synthesized from sucrose by the Lsc3Asp300Asn, were degraded most rapidly...... whilst levan produced by Lsc3 from raffinose least rapidly. BT1760 catalyzed finely at human body temperature (37°C) and in moderately acidic environment (pH 5-6) that is typical for the gut lumen. According to differential scanning fluorimetry, the Tm of the endo-levanase was 51.5°C. All tested levans...

  10. Integration of capillary electrophoresis with gold nanoparticle-based colorimetry.

    Science.gov (United States)

    Li, Tong; Wu, Zhenglong; Qin, Weidong

    2017-12-01

    A method integrating capillary electrophoresis (CE) and gold nanoparticle aggregation-based colorimetry (AuNP-ABC) was described. By using a dual-sheath interface, the running buffer was isolated from the colorimetric reaction solution so that CE and AuNP-ABC would not interfere with each other. The proof-of-concept was validated by assay of polyamidoamine (PAMAM) dendrimers that were fortified in human urine samples. The factors influencing the CE-AuNP-ABC performances were investigated and optimized. Under the optimal conditions, the dendrimers were separated within 8 min, with detection limits of 0.5, 1.2 and 2.6 μg mL -1 for PAMAM G1.0, G2.0 and G3.0, respectively. The sensitivity of CE-AuNP-ABC was comparable to or even better than those of liquid chromatography-fluorimetry and liquid chromatography-mass spectrometry. The results suggested that the proposed strategy can be applied to facile and quick determination of analytes of similar properties in complex matrices. Copyright © 2017 Elsevier B.V. All rights reserved.

  11. A Comparative Analysis of Uranium Ore using Laser Fluorimetric and gamma Spectrometry Techniques

    International Nuclear Information System (INIS)

    Madbouly, M.; Nassef, M. H.; El-Mongy, S.A.; Diab, A.M.

    2009-01-01

    A developed chemical separation method was used for the analysis of uranium in a standard U-ore (IAEA-RGU-1) by LASER fluorimetric technique. The non-destructive gamma assay technique was also applied to verify and compare the uranium content analyzed using laser technique. The results of the uranium analysis obtained by laser fluorimetry were found to be in the range of 360 - 420 μg/g with an average value of 390 μg/g. The bias between the measured and the certified value does not exceed 9.9%. For gamma-ray spectrometric analysis, the results of the measured uranium content were found to be in the range of 393.8 - 399.4 μg/g with an average value of 396.3 μg/g. The % difference in the case of γ- assay was 1.6 %. In general, the methods of analysis used in this study are applicable for a precise determination of uranium. It can be concluded that, laser analysis is preferred for assay of uranium ore due to the required small sample weight, the low time of sample preparation and cost of analysis.

  12. Interactions between radionuclides and organic colloids. Structure and reactivity of humic compounds; Interactions entre radionucleides et colloides organiques. Structure et reactivite des substances humiques

    Energy Technology Data Exchange (ETDEWEB)

    Plancque, G

    2001-09-01

    Humic compounds are the main organic colloids present in natural waters. These compounds can significantly modify the speciation of metals and control their properties, like migration, toxicity or bio-availability. It is thus important to study their speciation in conditions representative to those encountered in the natural environment. The aim of this work is to analyze the reactivity of these humic compounds. Two spectroscopic techniques have been used: the time-resolution laser spectro-fluorimetry, limited to the study of fluorescent elements, and the electro-spray source mass spectroscopy which requires the development of specific protocols for all elements of the periodic classification system. Europium, a fluorescent element analogue to trivalent actinides, has been chosen as test-metal for the intercomparison of both spectroscopic techniques. The first technique has permitted to determine the inorganic and organic speciation (spectra and lifetime of europium hydroxides and carbonates, and constants of interaction with humic acids, respectively). The limitations of this technique in the study of inorganic speciation has been evidenced. Humic compounds have a badly defined structure. The use of high-resolution mass spectroscopy has permitted to propose in a direct and experimental way, a molecular structure of aquatic fulvic acids in agreement with their known physico-chemical properties. (J.S.)

  13. Structure-Based Virtual Ligand Screening on the XRCC4/DNA Ligase IV Interface

    Science.gov (United States)

    Menchon, Grégory; Bombarde, Oriane; Trivedi, Mansi; Négrel, Aurélie; Inard, Cyril; Giudetti, Brigitte; Baltas, Michel; Milon, Alain; Modesti, Mauro; Czaplicki, Georges; Calsou, Patrick

    2016-03-01

    The association of DNA Ligase IV (Lig4) with XRCC4 is essential for repair of DNA double-strand breaks (DSBs) by Non-homologous end-joining (NHEJ) in humans. DSBs cytotoxicity is largely exploited in anticancer therapy. Thus, NHEJ is an attractive target for strategies aimed at increasing the sensitivity of tumors to clastogenic anticancer treatments. However the high affinity of the XRCC4/Lig4 interaction and the extended protein-protein interface make drug screening on this target particularly challenging. Here, we conducted a pioneering study aimed at interfering with XRCC4/Lig4 assembly. By Molecular Dynamics simulation using the crystal structure of the complex, we first delineated the Lig4 clamp domain as a limited suitable target. Then, we performed in silico screening of ~95,000 filtered molecules on this Lig4 subdomain. Hits were evaluated by Differential Scanning Fluorimetry, Saturation Transfer Difference - NMR spectroscopy and interaction assays with purified recombinant proteins. In this way we identified the first molecule able to prevent Lig4 binding to XRCC4 in vitro. This compound has a unique tripartite interaction with the Lig4 clamp domain that suggests a starting chemotype for rational design of analogous molecules with improved affinity.

  14. Mechanistic study of the interaction of uranyl ions with zirconium oxide and zirconium silicate; Etude mecanistique de l'interaction des ions uranyle avec l'oxyde et le silicate de zirconium

    Energy Technology Data Exchange (ETDEWEB)

    Lomenech, C

    2002-04-01

    This work deals with structural and thermodynamic studies of the sorption of uranyl species on zircon and zirconia. After determination of the specific areas, of the pH of the isoelectric points, and of the sorption site numbers, thermodynamic data were obtained using alpha spectrometry, for different uranyl concentrations, different background electrolytes (NaClO{sub 4} or KNO{sub 3}) and different ionic strengths. The structural identification of the surface complexes and sorption sites was carried out using several spectroscopies: XPS spectroscopy allowed a determination of the nature of the sorption sites ({identical_to}Zr-O- on zirconia and {identical_to}Si-O- on zircon). Whereas fluorescence decay measurements gave the number of surface species, the combined use of XPS spectroscopy and laser spectro-fluorimetry enabled us to correlate differences in bonding energies and emission wavelengths with differences in the nature of the background electrolyte or in the pH of sorption; DRIFT spectroscopy was a powerful tool for the determination of the presence of sorbed uranyl nitrate species. EXAFS results clearly showed a splitting of the equatorial oxygen atoms in two shells, corresponding to a polydentate, inner-sphere complex. EXAFS results also indicated strong similarities between dry samples and in situ experiments, which confirms the validity of all the spectroscopic measurements. Macroscopic thermodynamic data were then modeled using a surface complexation model (2 pK and constant capacitance models), the results of the structural study being used as constraints for the simulation code FITEQL. (author)

  15. Non-invasive and micro-destructive investigation of the Domus Aurea wall painting decorations.

    Science.gov (United States)

    Clementi, Catia; Ciocan, Valeria; Vagnini, Manuela; Doherty, Brenda; Tabasso, Marisa Laurenzi; Conti, Cinzia; Brunetti, Brunetto Giovanni; Miliani, Costanza

    2011-10-01

    The paper reports on the exploitation of an educated multi-technique analytical approach based on a wide non invasive step followed by a focused micro-destructive step, aimed at the minimally invasive identification of the pigments decorating the ceiling of the Gilded Vault of the Domus Aurea in Rome. The combination of elemental analysis with molecular characterization provided by X-ray fluorescence and UV-vis spectroscopies, respectively, allowed for the in situ non-invasive identification of a remarkable number of pigments, namely Egyptian blue, green earth, cinnabar, red ochre and an anthraquinonic lake. The study was completed with the Raman analysis of two bulk samples, in particular, SERS measurements allowed for the speciation of the anthraquinonic pigment. Elemental mapping by scanning electron microscopy-energy dispersive spectrometer combined with micro-fluorimetry on cross-section gave an insight into both the distribution of different blend of pigments and on the nature of the inorganic support of the red dye.

  16. Exploring the Trypanosoma brucei Hsp83 potential as a target for structure guided drug design.

    Directory of Open Access Journals (Sweden)

    Juan Carlos Pizarro

    Full Text Available Human African trypanosomiasis is a neglected parasitic disease that is fatal if untreated. The current drugs available to eliminate the causative agent Trypanosoma brucei have multiple liabilities, including toxicity, increasing problems due to treatment failure and limited efficacy. There are two approaches to discover novel antimicrobial drugs--whole-cell screening and target-based discovery. In the latter case, there is a need to identify and validate novel drug targets in Trypanosoma parasites. The heat shock proteins (Hsp, while best known as cancer targets with a number of drug candidates in clinical development, are a family of emerging targets for infectious diseases. In this paper, we report the exploration of T. brucei Hsp83--a homolog of human Hsp90--as a drug target using multiple biophysical and biochemical techniques. Our approach included the characterization of the chemical sensitivity of the parasitic chaperone against a library of known Hsp90 inhibitors by means of differential scanning fluorimetry (DSF. Several compounds identified by this screening procedure were further studied using isothermal titration calorimetry (ITC and X-ray crystallography, as well as tested in parasite growth inhibitions assays. These experiments led us to the identification of a benzamide derivative compound capable of interacting with TbHsp83 more strongly than with its human homologs and structural rationalization of this selectivity. The results highlight the opportunities created by subtle structural differences to develop new series of compounds to selectively target the Trypanosoma brucei chaperone and effectively kill the sleeping sickness parasite.

  17. Uranium occurence in California near Bucaramanga (Columbia)

    International Nuclear Information System (INIS)

    Heider Polania, J.

    1980-01-01

    The mining district of California, Bucaramanga, is on the west side of the Cordillera Oriental in the Santander massif region. The oldest rocks of the area form a complex of metamorphites and migmatites of the predevonic age. Amphibolite various types of paragneiss and orthogneiss are represented. Several stages of metamorphism can be documented in some rocks, as well as double anatexis. Triassic to jurassic quarz diorites and leukogranites show wide distribution. Porphyric rocks of granodioritic to granitic composition, to which the uranium mineralization is mainly bonded, intruded into the sediments of the lower cretaceous. Atomic absorption spectral analyses were carried out for the elements Cu, Zn and Li, as well as the uranium contents of some samples using fluorimetry. Uranium is primarily bonded to pitch blende and coffinite. The latter mostly occur in fine distribution grown in quarz and belong to the most recent mineralization phase. Autunite, meta-autunite, torbernite, meta-torbernite, zeunerite, meta-zeunerite and meta uranocircite detected as secondary uranium minerals. (orig./HP) [de

  18. A highly sensitive solid substrate room temperature phosphorimetry for carbaryl detection based on its activating effect on NaIO4 oxidizing fluorescein.

    Science.gov (United States)

    Liu, Jiaming; Huang, Qitong; Liu, Zhen-bo; Lin, Xiaofeng; Zhang, Li-Hong; Lin, Chang-Qing; Zheng, Zhi-Yong

    2014-11-01

    Fluorescein (HFin) could emit strong and stable room temperature phosphorescence (RTP) signal on polyamide membrane (PAM) using Pb(2+) as the ion perturber. Carbaryl could activate effect on NaIO4 oxidating HFin, which caused the RTP signal of the system to quench sharply. The phosphorescence intensity (ΔI p) of activating system higher 3.3 times (119.4/36.0) than that of non-activating system, and is directly proportional to the content of carbaryl. Thus, an activating solid substrate room temperature phosphorimetry (SSRTP) for carbaryl detection has been established. This sensitive (the limit of quantification (LOQ) was 2.0 × 10(-13) g mL(-1)), selective, simple and rapid method has been applied to determine trace carbaryl in water samples with the results consisting with those obtained by fluorimetry, showing its high accuracy. The apparent activation energy (E) and rate constant (k) of this activating reaction were 20.77 kJ mol(-1) and 1.85 × 10(-4) s(-1), respectively. Meanwhile, the mechanism of activating SSRTP for carbaryl detection was also discussed using infrared spectra (IR).

  19. Investigation of technology for the monitoring of UF6 mass flow in UF6 streams diluted with H2

    International Nuclear Information System (INIS)

    Baker, O.J.; Cooley, J.N.; Hewgley, W.A.; Moran, B.W.; Swindle, D.W. Jr.

    1986-12-01

    The applicability, availability, and effectiveness of gas flow meters are assessed as a means for verifying the mass flows of pure UF 6 streams diluted with a carrier gas. The initial survey identified the orifice, pitot tube, thermal, vortex shedding, and vortex precession (swirl) meters as promising for the intended use. Subsequent assessments of these flow meters revealed that two - the orifice meter and the pitot tube meter - are the best choices for the proposed applications: the first is recommended for low velocity gas, small diameter piping; the latter, for high velocity gas, large diameter piping. Final selection of the gas flow meters should be based on test loop evaluations in which the proposed meters are subjected to gas flows, temperatures, and pressures representative of those expected in service. Known instruments are evaluated that may be applicable to the measurement of uranium or UF 6 concentration in a UF 6 - H 2 process stream at an aerodynamic enrichment plant. Of the six procedures evaluated, four have been used for process monitoring in a UF 6 environment: gas mass spectrometry, infrared-ultraviolet-visible spectrophotometry, gas chromatography, and acoustic gas analysis. The remaining two procedures, laser fluorimetry and atomic absorption spectroscopy, would require significant development work before they could be used for process monitoring. Infrared-ultravioloet-visible spectrophotometry is judged to be the best procedure currently available to perform the required measurement

  20. Cyclic PaO2 oscillations assessed in the renal microcirculation: correlation with tidal volume in a porcine model of lung lavage.

    Science.gov (United States)

    Thomas, Rainer; Möllmann, Christian; Ziebart, Alexander; Liu, Tanghua; David, Matthias; Hartmann, Erik K

    2017-07-11

    Oscillations of the arterial partial pressure of oxygen induced by varying shunt fractions occur during cyclic alveolar recruitment within the injured lung. Recently, these were proposed as a pathomechanism that may be relevant for remote organ injury following acute respiratory distress syndrome. This study examines the transmission of oxygen oscillations to the renal tissue and their tidal volume dependency. Lung injury was induced by repetitive bronchoalveolar lavage in eight anaesthetized pigs. Cyclic alveolar recruitment was provoked by high tidal volume ventilation. Oscillations of the arterial partial pressure of oxygen were measured in real-time in the macrocirculation by multi-frequency phase fluorimetry and in the renal microcirculation by combined white-light spectrometry and laser-Doppler flowmetry during tidal volume down-titration. Significant respiratory-dependent oxygen oscillations were detected in the macrocirculation and transmitted to the renal microcirculation in a substantial extent. The amplitudes of these oscillations significantly correlate to the applied tidal volume and are minimized during down-titration. In a porcine model oscillations of the arterial partial pressure of oxygen are induced by cyclic alveolar recruitment and transmitted to the renal microcirculation in a tidal volume-dependent fashion. They might play a role in organ crosstalk and remote organ damage following lung injury.

  1. Elucidating the weak protein-protein interaction mechanisms behind the liquid-liquid phase separation of a mAb solution by different types of additives.

    Science.gov (United States)

    Wu, Guoliang; Wang, Shujing; Tian, Zhou; Zhang, Ning; Sheng, Han; Dai, Weiguo; Qian, Feng

    2017-11-01

    Liquid-liquid phase separation (LLPS) has long been observed during the physical stability investigation of therapeutic protein formulations. The buffer conditions and the presence of various excipients are thought to play important roles in the formulation development of monoclonal antibodies (mAbs). In this study, the effects of several small-molecule excipients (histidine, alanine, glycine, sodium phosphate, sodium chloride, sorbitol and sucrose) with diverse physical-chemical properties on LLPS of a model IgG1 (JM2) solutions were investigated by multiple techniques, including UV-vis spectroscopy, circular dichroism, differential scanning calorimetry/fluorimetry, size exclusion chromatography and dynamic light scattering. The LLPS of JM2 was confirmed to be a thermodynamic equilibrium process with no structural changes or irreversible aggregation of proteins. Phase diagrams of various JM2 formulations were constructed, suggesting that the phase behavior of JM2 was dependent on the solution pH, ionic strength and the presence of other excipients such as glycine, alanine, sorbitol and sucrose. Furthermore, we demonstrated that for this mAb, the interaction parameter (k D ) determined at low protein concentration appeared to be a good predictor for the occurrence of LLPS at high concentration. Copyright © 2017 Elsevier B.V. All rights reserved.

  2. Immunobiological effect of bitemporal exposure of rabbits to microwaves

    Energy Technology Data Exchange (ETDEWEB)

    Bogolyubov, V.M.; Pershin, S.B.; Frenkel' , I.D.; Sidorov, V.D.; Galenchik, A.I.; Ponomarev, Yu.T.; Bobkova, A.S.; Kuz' min, S.N.; Moshiashvili, I.Ya.; Kozlova, N.N.; Korovkina, E.G.; Agibalov, Yu.V.

    1987-01-01

    The authors investigate the effects of microwave radiation on the immunological behavior of the thyroid and various hormones of rabbits. Irradiation was carried out on the heads of the animals. They were then divided into four groups depending on the period of exposure. The number of hemolysis-forming cells against sheep red blood cells and the concentration of serum immunoglobulins were determined. Levels of TSH, triiodothyronine, thyroxine, testosterone and prostaglandins in serum or blood plasma were determined by radioimmunoassay and the concentration of total 11-hydroxycorticosteroids in the adrenals and plasma were determined by fluorimetry. Microwave radiation of the temporo-parietal region of the head was found to decrease the number of background hemolysis-forming cells. An increase in glucocorticoid function was recorded. Thyroid function was depressed. The plasma 11-hydroxycorticosteroid level was significantly raised. It is concluded in general that microwave irradiation leads to activation of the hypothalamo-hypophyseo-adrenal system with consequent enhancement of the glucocorticoid function of the adrenal cortex and depression of thyroid function.

  3. Full-length model of the human galectin-4 and insights into dynamics of inter-domain communication

    Science.gov (United States)

    Rustiguel, Joane K.; Soares, Ricardo O. S.; Meisburger, Steve P.; Davis, Katherine M.; Malzbender, Kristina L.; Ando, Nozomi; Dias-Baruffi, Marcelo; Nonato, Maria Cristina

    2016-09-01

    Galectins are proteins involved in diverse cellular contexts due to their capacity to decipher and respond to the information encoded by β-galactoside sugars. In particular, human galectin-4, normally expressed in the healthy gastrointestinal tract, displays differential expression in cancerous tissues and is considered a potential drug target for liver and lung cancer. Galectin-4 is a tandem-repeat galectin characterized by two carbohydrate recognition domains connected by a linker-peptide. Despite their relevance to cell function and pathogenesis, structural characterization of full-length tandem-repeat galectins has remained elusive. Here, we investigate galectin-4 using X-ray crystallography, small- and wide-angle X-ray scattering, molecular modelling, molecular dynamics simulations, and differential scanning fluorimetry assays and describe for the first time a structural model for human galectin-4. Our results provide insight into the structural role of the linker-peptide and shed light on the dynamic characteristics of the mechanism of carbohydrate recognition among tandem-repeat galectins.

  4. The selective uptake of uranium and thorium from the environment by some vegetables

    International Nuclear Information System (INIS)

    Yusof, A.M.; Ghazali, Z.; Abdul-Rahman, S.; Sharif, J.

    1991-01-01

    An attempt was made to establish baseline information on environmental pollution in locally-grown vegetables by uranium and thorium. Lowland and highland species together with soil and fertilizer samples were collected and analyzed using fluorimetry, spectrophotometry and delayed neutron counting techniques. All leafy vegetables observed showed high uranium and thorium uptake especially those grown in the lowlands. Those grown in the highlands reflected no direct relationship in uranium and thorium contents. Several species common in both sampling areas exhibited direct relationship between these two elements making them as potential bio-indicators. Figures calculated for fruit-type and leafy vegetables were not only comparatively low but bore no direct correlation between the two elements. The use of phosphate-based fertilizers on some of the leafy species in the lowlands did not enhance the uptake of these elements in spite of the higher uranium and thorium contents in soil samples from the lowlands, between 20-85 μg/g for uranium and 43-226 μg/g thorium compared to about 13-20 μg/g and 35-55 μg/g respectively for soil samples in the highlands. Statistical analysis was done to substantiate these findings. Climatic conditions were also taken into account as one of the factors affecting selective uptake of these elements in the vegetables

  5. A Systematic Protein Refolding Screen Method using the DGR Approach Reveals that Time and Secondary TSA are Essential Variables.

    Science.gov (United States)

    Wang, Yuanze; van Oosterwijk, Niels; Ali, Ameena M; Adawy, Alaa; Anindya, Atsarina L; Dömling, Alexander S S; Groves, Matthew R

    2017-08-24

    Refolding of proteins derived from inclusion bodies is very promising as it can provide a reliable source of target proteins of high purity. However, inclusion body-based protein production is often limited by the lack of techniques for the detection of correctly refolded protein. Thus, the selection of the refolding conditions is mostly achieved using trial and error approaches and is thus a time-consuming process. In this study, we use the latest developments in the differential scanning fluorimetry guided refolding approach as an analytical method to detect correctly refolded protein. We describe a systematic buffer screen that contains a 96-well primary pH-refolding screen in conjunction with a secondary additive screen. Our research demonstrates that this approach could be applied for determining refolding conditions for several proteins. In addition, it revealed which "helper" molecules, such as arginine and additives are essential. Four different proteins: HA-RBD, MDM2, IL-17A and PD-L1 were used to validate our refolding approach. Our systematic protocol evaluates the impact of the "helper" molecules, the pH, buffer system and time on the protein refolding process in a high-throughput fashion. Finally, we demonstrate that refolding time and a secondary thermal shift assay buffer screen are critical factors for improving refolding efficiency.

  6. Synergistic Effect of MiR-146a Mimic and Cetuximab on Hepatocellular Carcinoma Cells

    Directory of Open Access Journals (Sweden)

    Suning Huang

    2014-01-01

    Full Text Available Previously, we found that the expression of microRNA-146a (miR-146a was downregulated in hepatocellular carcinoma (HCC formalin-fixed paraffin-embedded (FFPE tissues compared to the adjacent noncancerous hepatic tissues. In the current study, we have explored the in vitro effect of miR-146a on the malignant phenotypes of HCC cells. MiR-146a mimic could suppress cell growth and increase cellular apoptosis in HCC cell lines HepG2, HepB3, and SNU449, as assessed by spectrophotometry, fluorimetry, and fluorescence microscopy, respectively. Furthermore, western blot showed that miR-146a mimic downregulated EGFR, ERK1/2, and stat5 signalings. These effects were less potent compared to that of a siRNA targeting EGFR, a known target gene of miR-146a. Moreover, miR-146a mimic could enhance the cell growth inhibition and apoptosis induction impact of various EGFR targeting agents. The most potent combination was miR-146a mimic with cetuximab, presenting a synergistic effect. In conclusion, miR-146a plays a vital role in the cell growth and apoptosis of HCC cells and inducing miR-146a level might be a critical targeted molecular therapy strategy for HCC.

  7. Laser fluorimetric analysis of uranium in water from Vishakhapatnam and estimation of health risk

    International Nuclear Information System (INIS)

    Bhangare, R.C.; Tiwari, M.; Ajmal, P.Y.; Sahu, S.K.; Pandit, G.G.

    2013-01-01

    Uranium is a naturally occurring radioactive element that is both radiologically and chemically toxic. The presence of uranium in the aquatic environment is due to the leaching from natural deposits, release in mill tailings, the combustion of coal and other fuels, and the use of phosphate fertilizers that contain uranium. Intake of uranium through air and water is normally low, but in circumstances in which uranium is present in a drinking water source, the majority of intake can be through drinking water route. The uranium concentrations in ground water samples from Vishakhapatnam, India were estimated using laser fluorimetric technique and were observed to range from 0.6 to 12.3 ppb. The laser fluorimetry technique was found to be an excellent tool for direct measurement of uranium concentration in water samples at ultra-trace levels. The annual effective dose, cumulative dose for 70 years and the lifetime excess cancer risk from drinking of this water were calculated. The risks were low averaging only 10.6 x 10 -6 as none of the samples were observed to exceed the WHO recommended uranium concentration limit of 30 ppb. (author)

  8. Preparation data of the bromodomains BRD3(1, BRD3(2, BRD4(1, and BRPF1B and crystallization of BRD4(1-inhibitor complexes

    Directory of Open Access Journals (Sweden)

    Martin Hügle

    2016-06-01

    Full Text Available This article presents detailed purification procedures for the bromodomains BRD3(1, BRD3(2, BRD4(1, and BRPF1B. In addition we provide crystallization protocols for apo BRD4(1 and BRD4(1 in complex with numerous inhibitors. The protocols described here were successfully applied to obtain affinity data by isothermal titration calorimetry (ITC and by differential scanning fluorimetry (DSF as well as structural characterizations of BRD4(1 inhibitor complexes (PDB codes: PDB: 4LYI, PDB: 4LZS, PDB: 4LYW, PDB: 4LZR, PDB: 4LYS, PDB: 5D24, PDB: 5D25, PDB: 5D26, PDB: 5D3H, PDB: 5D3J, PDB: 5D3L, PDB: 5D3N, PDB: 5D3P, PDB: 5D3R, PDB: 5D3S, PDB: 5D3T. These data have been reported previously and are discussed in more detail elsewhere [1,2].

  9. Metabolites of saxitoxin analogues in bivalves contaminated by Gymnodinium catenatum.

    Science.gov (United States)

    Vale, Paulo

    2010-01-01

    Bivalve metabolites of saxitoxin analogues, not present in microalgae, were recently described as an important toxin fraction in mussels contaminated by Alexandrium tamarense. These possess very low fluorescence, and require mass spectrometry detection. HILIC-MS was implemented to look for these metabolites in bivalves contaminated during Gymnodinium catenatum blooms at the Portuguese coast. The presence of M1 was tentatively identified in several bivalves, ranging from estuarine (Mytilus galloprovinciallis, Cerastoderma edule and Ruditapes decussatus) to oceanic habitat (Donax trunculus and Ensis spp.). It was hypothesized that M1 could contribute to an important fraction of the profile of STX analogues. M1 was more abundant in estuarine bivalves that retain longer PSP toxins, in the following order: mussels>cockles>clams. These data highlight that the study by fluorimetry alone of the carbamoyl, N-sulfocarbamoyl, and decarbamoyl families is manifestly insufficient to fully understand toxin dynamics in bivalves feeding on G. catenatum without a proper study of hydroxybenzoate and hydroxylated M-toxins.

  10. Report on intercomparisons S-14, S-15, and S-16 of the determination of uranium and thorium in thorium ores

    International Nuclear Information System (INIS)

    Pszonicki, L.; Hanna, A.N.; Suschny, O.

    1983-06-01

    Twenty-nine laboratories from 18 countries took part in this intercomparison, organized by the IAEA's Analytical Quality Control Service, to help laboratories engaged in this task to check the reliability of their results. An additional aim was to establish the concentrations of thorium and uranium in three large batches of thorium ores and certifying them as reference materials. The evaluation was based on 438 individual results (108 laboratory means) for thorium, and on 412 individual results (106 laboratory means) for uranium. The number of laboratory means per element and per sample varied from 34 to 38. The methods most frequently used in the determination of both elements were neutron activation analysis and radiometry. They were followed by spectrophotometry and X-ray fluorescence analysis for thorium and by fluorimetry, X-ray fluorescence analysis and spectrophotometry for uranium determination, respectively. The relative uncertainty of all computed overall medians which were used as the best estimations of true values, does not exceed +-10% and +-5% for the concentration values below and above 0.1%, respectively

  11. HFE gene variants and iron-induced oxygen radical generation in idiopathic pulmonary fibrosis.

    Science.gov (United States)

    Sangiuolo, Federica; Puxeddu, Ermanno; Pezzuto, Gabriella; Cavalli, Francesco; Longo, Giuliana; Comandini, Alessia; Di Pierro, Donato; Pallante, Marco; Sergiacomi, Gianluigi; Simonetti, Giovanni; Zompatori, Maurizio; Orlandi, Augusto; Magrini, Andrea; Amicosante, Massimo; Mariani, Francesca; Losi, Monica; Fraboni, Daniela; Bisetti, Alberto; Saltini, Cesare

    2015-02-01

    In idiopathic pulmonary fibrosis (IPF), lung accumulation of excessive extracellular iron and macrophage haemosiderin may suggest disordered iron homeostasis leading to recurring microscopic injury and fibrosing damage. The current study population comprised 89 consistent IPF patients and 107 controls. 54 patients and 11 controls underwent bronchoalveolar lavage (BAL). Haemosiderin was assessed by Perls' stain, BAL fluid malondialdehyde (MDA) by high-performance liquid chromatography, BAL cell iron-dependent oxygen radical generation by fluorimetry and the frequency of hereditary haemochromatosis HFE gene variants by reverse dot blot hybridisation. Macrophage haemosiderin, BAL fluid MDA and BAL cell unstimulated iron-dependent oxygen radical generation were all significantly increased above controls (pHFE allelic variants was markedly higher in IPF compared with controls (40.4% versus 22.4%, OR 2.35, p=0.008) and was associated with higher iron-dependent oxygen radical generation (HFE variant 107.4±56.0, HFE wild type (wt) 59.4±36.4 and controls 16.7±11.8 fluorescence units per 10(5) BAL cells; p=0.028 HFE variant versus HFE wt, p=0.006 HFE wt versus controls). The data suggest iron dysregulation associated with HFE allelic variants may play an important role in increasing susceptibility to environmental exposures, leading to recurring injury and fibrosis in IPF. Copyright ©ERS 2015.

  12. Identification of rare earth elements by their fluorescence complexes with tetracycline

    International Nuclear Information System (INIS)

    Zanotti-Cavazzoni, J.C.; Boveda V, J.C.; Abrao, A.

    1994-01-01

    A procedure for identification of rare earth elements (REE) based on the complex formation with tetracycline (TC) and visual observation under ultraviolet light is described. One micro drop of REE chloride is placed over the filter paper impregnated with tetracycline and previously dried before use. After dried (30-60 O C) the paper is examined under the UV light. In a second procedure for the identification of mixture of REE a drop of the analyte is added over a filter paper strip impregnated with TC followed by a chromatographic development using a 2:1:1:1 (vol/vol) ethanol, acetone, ether and 1% acetic acid mixture of pH 5,8. After dried, the strip is observed under UV light. Sc, Y, La, Dy, Ho and Lu exhibited high yellow fluorescence; Gd, Tm and Yb a weak yellow; Ce an intense violet; Pr, ND, Sm and Tb violet: Eu a high red fluorescence and Er a violet brown wish, not well defined. It is possible to identify for instance 0.3 ug Ce (concentration 6 mg/L) and 0.2 ug Er (concentration 6 mg/L). Preliminary experiments indicated that those REE-TC chelates can be used for the identification and determination of the REE by spectro fluorimetry. (author). 30 refs

  13. Mechanistic study of the interaction of uranyl ions with zirconium oxide and zirconium silicate

    International Nuclear Information System (INIS)

    Lomenech, C.

    2002-04-01

    This work deals with structural and thermodynamic studies of the sorption of uranyl species on zircon and zirconia. After determination of the specific areas, of the pH of the isoelectric points, and of the sorption site numbers, thermodynamic data were obtained using alpha spectrometry, for different uranyl concentrations, different background electrolytes (NaClO 4 or KNO 3 ) and different ionic strengths. The structural identification of the surface complexes and sorption sites was carried out using several spectroscopies: XPS spectroscopy allowed a determination of the nature of the sorption sites (≡Zr-O- on zirconia and ≡Si-O- on zircon). Whereas fluorescence decay measurements gave the number of surface species, the combined use of XPS spectroscopy and laser spectro-fluorimetry enabled us to correlate differences in bonding energies and emission wavelengths with differences in the nature of the background electrolyte or in the pH of sorption; DRIFT spectroscopy was a powerful tool for the determination of the presence of sorbed uranyl nitrate species. EXAFS results clearly showed a splitting of the equatorial oxygen atoms in two shells, corresponding to a polydentate, inner-sphere complex. EXAFS results also indicated strong similarities between dry samples and in situ experiments, which confirms the validity of all the spectroscopic measurements. Macroscopic thermodynamic data were then modeled using a surface complexation model (2 pK and constant capacitance models), the results of the structural study being used as constraints for the simulation code FITEQL. (author)

  14. A comprehensive Radiological Investigation for the Industrial Helwan City Environment

    International Nuclear Information System (INIS)

    Wafaa, F.; El-Tahawy, M.S.; El-Mongy, S.A.; Sayed, S.A.

    2000-01-01

    The non nuclear process industries supply a considerable contribution to the radiological pollution of the environment. A comprehensive radiological studies for determining radiation levels and radionuclides distribution in the environment of Helwan city (the major industrial region in Cairo) are carried out. The radioactivity concentrations of Ra-226, Th-232, K-40 and the man made Cs-137 in samples of soil plants, ores and products collected from and around some of Helwan factories were analyzed using high resolution gamma spectrometers. A bad correlation between the observed 137 Cs and 40 K was found. The collected water and factories waste water samples were also analyzed by laser fluorimetry. The absorbed dose rates due to the natural radioactivity in the soil one meter above the ground and the radium equivalent activities are calculated and presented. The representative external hazard index values for the soil samples to be used as construction materials are also estimated and given. The results obtained are considered as reference values to be used in the case of emergency action and as baseline data for drawing a radiological map of the city

  15. Towards an interpretation of the mechanism of the actinides(III)/lanthanides(III) separation by synergistic solvent extraction with nitrogen-containing polydendate ligands; Vers une interpretation des mecanismes de la separation actinides(III)/lanthanides(III) par extraction liquide-liquide synergique impliquant des ligands polyazotes

    Energy Technology Data Exchange (ETDEWEB)

    Francois, N. [CEA/VALRHO - site de Marcoule, Dept. de Recherche en Retraitement et en Vitrification, (DRRV), 30 - Marcoule (France); Universite Henri Poincare, 54 - Vandoeuvre-les-Nancy (France)

    2000-07-01

    In the field of the separation of long-lived radionuclides from the wastes produced by nuclear fuel reprocessing, aromatic nitrogen-containing polydendate ligands are potential candidates for the selective extraction, alone or in synergistic mixture with acidic extractants, of trivalent actinides from trivalent lanthanides. The first part of this work deals with the complexation of trivalent f cations with various nitrogen-containing ligands (poly-pyridine analogues). Time-resolved laser-induced fluorimetry (TRLIF) and UV-visible spectrophotometry were used to determine the nature and evaluate the stability of each complex. Among the ligands studied, the least basic Me-Btp proved to be highly selective towards americium(III) in acidic solution. In the second part, two synergistic systems (nitrogen-containing polydendate ligand and lipophilic carboxylic acid) are studied and compared in regard to the extraction and separation of lanthanides(III) and actinides(III). TRLIF and gamma spectrometry allowed the nature of the extracted complexes and the optimal conditions of efficiency of both systems to be determined. Comparison between these different studies showed that the selectivity of complexation of trivalent f cations by a given nitrogen-containing polydendate ligand could not always be linked to the Am(III)Eu(III) selectivity reached in synergistic extraction. The latter depends on the 'balance' between the acid-basic properties on the one hand, and on the hard-soft characteristics on the other hand, of both components of synergistic system. (author)

  16. The Axl kinase domain in complex with a macrocyclic inhibitor offers first structural insights into an active TAM receptor kinase.

    Science.gov (United States)

    Gajiwala, Ketan S; Grodsky, Neil; Bolaños, Ben; Feng, Junli; Ferre, RoseAnn; Timofeevski, Sergei; Xu, Meirong; Murray, Brion W; Johnson, Ted W; Stewart, Al

    2017-09-22

    The receptor tyrosine kinase family consisting of Tyro3, Axl, and Mer (TAM) is one of the most recently identified receptor tyrosine kinase families. TAM receptors are up-regulated postnatally and maintained at high levels in adults. They all play an important role in immunity, but Axl has also been implicated in cancer and therefore is a target in the discovery and development of novel therapeutics. However, of the three members of the TAM family, the Axl kinase domain is the only one that has so far eluded structure determination. To this end, using differential scanning fluorimetry and hydrogen-deuterium exchange mass spectrometry, we show here that a lower stability and greater dynamic nature of the Axl kinase domain may account for its poor crystallizability. We present the first structural characterization of the Axl kinase domain in complex with a small-molecule macrocyclic inhibitor. The Axl crystal structure revealed two distinct conformational states of the enzyme, providing a first glimpse of what an active TAM receptor kinase may look like and suggesting a potential role for the juxtamembrane region in enzyme activity. We noted that the ATP/inhibitor-binding sites of the TAM members closely resemble each other, posing a challenge for the design of a selective inhibitor. We propose that the differences in the conformational dynamics among the TAM family members could potentially be exploited to achieve inhibitor selectivity for targeted receptors. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

  17. Label-free DNA quantification via a 'pipette, aggregate and blot' (PAB) approach with magnetic silica particles on filter paper.

    Science.gov (United States)

    Li, Jingyi; Liu, Qian; Alsamarri, Hussein; Lounsbury, Jenny A; Haversitick, Doris M; Landers, James P

    2013-03-07

    Reliable measurement of DNA concentration is essential for a broad range of applications in biology and molecular biology, and for many of these, quantifying the nucleic acid content is inextricably linked to obtaining optimal results. In its most simplistic form, quantitative analysis of nucleic acids can be accomplished by UV-Vis absorbance and, in more sophisticated format, by fluorimetry. A recently reported new concept, the 'pinwheel assay', involves a label-free approach for quantifying DNA through aggregation of paramagnetic beads in a rotating magnetic field. Here, we describe a simplified version of that assay adapted for execution using only a pipet and filter paper. The 'pipette, aggregate, and blot' (PAB) approach allows DNA to induce bead aggregation in a pipette tip through exposure to a magnetic field, followed by dispensing (blotting) onto filter paper. The filter paper immortalises the extent of aggregation, and digital images of the immortalized bead conformation, acquired with either a document scanner or a cell phone camera, allows for DNA quantification using a noncomplex algorithm. Human genomic DNA samples extracted from blood are quantified with the PAB approach and the results utilized to define the volume of sample used in a PCR reaction that is sensitive to input mass of template DNA. Integrating the PAB assay with paper-based DNA extraction and detection modalities has the potential to yield 'DNA quant-on-paper' devices that may be useful for point-of-care testing.

  18. Combining Fungicides and Prospective NPR1-Based "Just-in-Time" Immunomodulating Chemistries for Crop Protection.

    Science.gov (United States)

    Kuai, Xiahezi; Barraco, Charles; Després, Charles

    2017-01-01

    Each year, crop yield is lost to weeds competing for resources, insect herbivory and diseases caused by pathogens. To thwart these insults and preserve yield security and a high quality of traits, conventional agriculture makes use of improved cultivars combined with fertilizer and agrochemical applications. However, given that regulatory bodies and consumers are demanding environmentally safer agrochemicals, while at the same time resistance to agrochemicals is mounting, it is crucial to adopt a "holistic" approach to agriculture by not excluding any number of management tools at our disposal. One such tool includes chemicals that stimulate plant immunity. The development of this particular type of alternative crop protection strategy has been of great interest to us. We have approached this paradigm by studying plant immunity, specifically systemic acquired resistance (SAR). The deployment of SAR immunity requires the production by the crop plant of an endogenous small molecule metabolite called salicylic acid (SA). Furthermore, immunity can only be deployed if SA can bind to its receptor and activate the genes responsible for the SAR program. The key receptor for SAR is a transcription coactivator called NPR1. Since discovering this NPR1-SA receptor-ligand pair, we have embarked on a journey to develop novel chemistries capable of deploying SAR in the field. The journey begins with the development of a scalable assay to identify these novel chemistries. One such assay, presented here, is based on differential scanning fluorimetry technology and demonstrates that NPR1 is destabilized by binding to SA.

  19. The effect of ionizing irradiation on the catecholamine levels in pituitary gland of ewes

    International Nuclear Information System (INIS)

    Pastorova, B.; Stanikova, A.; Maracek, I.; Danko, J.

    2004-01-01

    The changes were studied in the levels of catecholamines and L-DOPA in the pituitary gland of sheep after all-body irradiation with 60 Co at the total dose of 6.7 Gy for seven days. The power input per hour of irradiation source was 0.039 Gy. The catecholamines were determined after having been isolated from the tissues and determined by the method of spectral fluorimetry. After all-body exposition to gamma-radiation L-DOPA dropped in pituitary gland in comparison with the control group of sheep most significantly by 66.7% (P < 0,001). A similar drop like in the case of adrenaline was also observed in epinephrine (P < 0,05). On basis of the results we presume that the all-body irradiation of sheep by results a significant decrease in the content of catecholamines in pituitary gland, which is probably in relation with failure of synthesis and degradation of catecholamines and with total organism injury. (authors)

  20. Functional characterisation of Burkholderia pseudomallei biotin protein ligase: A toolkit for anti-melioidosis drug development.

    Science.gov (United States)

    Bond, Thomas E H; Sorenson, Alanna E; Schaeffer, Patrick M

    2017-06-01

    Burkholderia pseudomallei (Bp) is the causative agent of melioidosis. The bacterium is responsible for 20% of community-acquired sepsis cases and 40% of sepsis-related mortalities in northeast Thailand, and is intrinsically resistant to aminoglycosides, macrolides, rifamycins, cephalosporins, and nonureidopenicillins. There is no vaccine and its diagnosis is problematic. Biotin protein ligase (BirA) which is essential for fatty acid synthesis has been proposed as a drug target in bacteria. Very few bacterial BirA have been characterized, and a better understanding of these enzymes is necessary to further assess their value as drug targets. BirA within the Burkholderia genus have not yet been investigated. We present for the first time the cloning, expression, purification and functional characterisation of the putative Bp BirA and orthologous B. thailandensis (Bt) biotin carboxyl carrier protein (BCCP) substrate. A GFP-tagged Bp BirA was produced and applied for the development of a high-throughput (HT) assay based on our differential scanning fluorimetry of GFP-tagged proteins (DSF-GTP) principle as well as an electrophoretic mobility shift assay. Our biochemical data in combination with the new HT DSF-GTP and biotinylation activity assay could facilitate future drug screening efforts against this drug-resistant organism. Copyright © 2017 Elsevier GmbH. All rights reserved.

  1. Assessment of Pseudomonas aeruginosa N5,N10-methylenetetrahydrofolate dehydrogenase-cyclohydrolase as a potential antibacterial drug target.

    Directory of Open Access Journals (Sweden)

    Thomas C Eadsforth

    Full Text Available The bifunctional enzyme methylenetetrahydrofolate dehydrogenase - cyclohydrolase (FolD is identified as a potential drug target in Gram-negative bacteria, in particular the troublesome Pseudomonas aeruginosa. In order to provide a comprehensive and realistic assessment of the potential of this target for drug discovery we generated a highly efficient recombinant protein production system and purification protocol, characterized the enzyme, carried out screening of two commercial compound libraries by differential scanning fluorimetry, developed a high-throughput enzyme assay and prosecuted a screening campaign against almost 80,000 compounds. The crystal structure of P. aeruginosa FolD was determined at 2.2 Å resolution and provided a template for an assessment of druggability and for modelling of ligand complexes as well as for comparisons with the human enzyme. New FolD inhibitors were identified and characterized but the weak levels of enzyme inhibition suggest that these compounds are not optimal starting points for future development. Furthermore, the close similarity of the bacterial and human enzyme structures suggest that selective inhibition might be difficult to attain. In conclusion, although the preliminary biological data indicates that FolD represents a valuable target for the development of new antibacterial drugs, indeed spurred us to investigate it, our screening results and structural data suggest that this would be a difficult enzyme to target with respect to developing the appropriate lead molecules required to underpin a serious drug discovery effort.

  2. Evaluating the effects of buffer conditions and extremolytes on thermostability of granulocyte colony-stimulating factor using high-throughput screening combined with design of experiments.

    Science.gov (United States)

    Ablinger, Elisabeth; Hellweger, Monika; Leitgeb, Stefan; Zimmer, Andreas

    2012-10-15

    In this study, we combined a high-throughput screening method, differential scanning fluorimetry (DSF), with design of experiments (DoE) methodology to evaluate the effects of several formulation components on the thermostability of granulocyte colony stimulating factor (G-CSF). First we performed a primary buffer screening where we tested thermal stability of G-CSF in different buffers, pH values and buffer concentrations. The significance of each factor and the two-way interactions between them were studied by multivariable regression analysis. pH was identified as most critical factor regarding thermal stability. The most stabilizing buffer, sodium glutamate, and sodium acetate were determined for further investigations. Second we tested the effect of 6 naturally occurring extremolytes (trehalose, sucrose, ectoine, hydroxyectoine, sorbitol, mannitol) on the thermal stability of G-CSF, using a central composite circumscribed design. At low pH (3.8) and low buffer concentration (5 mM) all extremolytes led to a significant increase in thermal stability except the addition of ectoine which resulted in a strong destabilization of G-CSF. Increasing pH and buffer concentration led to an increase in thermal stability with all investigated extremolytes. The described systematic approach allowed to create a ranking of stabilizing extremolytes at different buffer conditions. Copyright © 2012. Published by Elsevier B.V.

  3. Study of OH● Radicals in Human Serum Blood of Healthy Individuals and Those with Pathological Schizophrenia

    Directory of Open Access Journals (Sweden)

    Wolfgang Linert

    2011-01-01

    Full Text Available The human body is constantly under attack from free radicals that occur as part of normal cell metabolism, and by exposure to environmental factors such as UV light, cigarette smoke, environmental pollutants and gamma radiation. The resulting “Reactive Oxygen Species” (ROS circulate freely in the body with access to all organs and tissues, which can have serious repercussions throughout the body. The body possesses a number of mechanisms both to control the production of ROS and to cope with free radicals in order to limit or repair damage to tissues. Overproduction of ROS or insufficient defense mechanisms leads to a dangerous disbalance in the organism. Thereby several pathomechanisms implicated in over 100 human diseases, e.g., cardiovascular disease, cancer, diabetes mellitus, physiological disease, aging, etc., can be induced. Thus, a detailed investigation on the quantity of oxygen radicals, such as hydroxyl radicals (OH● in human serum blood, and its possible correlation with antioxidant therapy effects, is highly topical. The subject of this study was the influence of schizophrenia on the amount of OH● in human serum blood. The radicals were detected by fluorimetry, using terephthalic acid as a chemical trap. For all experiments the serum blood of healthy people was used as a control group.

  4. Process, product, and waste-stream monitoring with fiber optics

    International Nuclear Information System (INIS)

    Milanovich, F.P.; Hirschfeld, T.

    1983-07-01

    Fiber optic technology, motivated by communications and defense applications, has advanced significantly the past ten years. In particular, advances have been made in visible radiation transmission efficiency with concurrent reductions in fiber size, weight, and cost. Researchers at the Lawrence Livermore National Laboratory (LLNL) coupled these advances in fiber optic technology with analytical fluorescence analysis to establish a new technology - remote fiber fluorimetry (RFF). Laser-based RFF offers the potential to measure and monitor from one central and remote laboratory, on-line, and in near real time, trace (ppM) to substantial (g/L) concentrations of selected chemical species in typical process, product, and waste streams. The fluorimeter consists of a fluorescence or Raman spectrometer; unique coupling optics that separates input excitation (laser) radiation from return (fluorescence) radiation; a fiber optic cable; and an optrode - a terminal that interfaces the fiber to the measurement point, which is designed to respond quantitatively to a particular chemical species. At LLNL, research is underway into optrodes that measure pressure, temperature, and pH and those that detect and quantify various actinides, sulfates, inorganic chloride, hydrogen sulfide, aldehydes, and alcohols

  5. Evaluation of the effective equivalent dose in the general public due to the discharge of uranium in groundwater

    International Nuclear Information System (INIS)

    Gordon, A.M.P.L.; Jacomino, V.M.F.

    1989-12-01

    Some facilities available at IPEN-CNEN/SP may discharge uranium in their liquid effluents. The uranium contents of these effluents are analyzed by photometry or fluorimetry, and according to the results obtained a decision is made, by the Environmental Monitoring Division, upon their discharge to the environment. In 1988 a total activity of 3.66x10 9 Bq of uranium was discharge in a volume of approximately 30 m 3 . The effective equivalent dose in the general public was evaluated by making a conservative assumption that all the liquid effluents containing uranium are discharged directly to the soil reaching the groundwater. The dose calculation was carried out by using a generic model which described the transport of radionuclides in the groundwater. In order to be conservative it was also assumed that the critical pathway is the direct in gestion of water through hypothetical wells around the Institute. Conservative assumptions were also made in the characterization of the local aquifer parameters such as vertical and longitudinal dispersivity, effective porosity of the soil, hydraulic conductivity etc., in roder to overestimate the effective equivalent dose. The result obtained, of 5.3x10 -10 mSv/a is far below the dose limit for the public adopted by the Radiological Protection Board. The derived limit for the discharge was also evaluated, using the same model, giving a result of 3.6x10 13 Bq/a. (author) [pt

  6. Resolving Hot Spots in the C-Terminal Dimerization Domain that Determine the Stability of the Molecular Chaperone Hsp90

    Science.gov (United States)

    Reimann, Sven; Smits, Sander H. J.; Schmitt, Lutz; Groth, Georg; Gohlke, Holger

    2014-01-01

    Human heat shock protein of 90 kDa (hHsp90) is a homodimer that has an essential role in facilitating malignant transformation at the molecular level. Inhibiting hHsp90 function is a validated approach for treating different types of tumors. Inhibiting the dimerization of hHsp90 via its C-terminal domain (CTD) should provide a novel way to therapeutically interfere with hHsp90 function. Here, we predicted hot spot residues that cluster in the CTD dimerization interface by a structural decomposition of the effective energy of binding computed by the MM-GBSA approach and confirmed these predictions using in silico alanine scanning with DrugScorePPI. Mutation of these residues to alanine caused a significant decrease in the melting temperature according to differential scanning fluorimetry experiments, indicating a reduced stability of the mutant hHsp90 complexes. Size exclusion chromatography and multi-angle light scattering studies demonstrate that the reduced stability of the mutant hHsp90 correlates with a lower complex stoichiometry due to the disruption of the dimerization interface. These results suggest that the identified hot spot residues can be used as a pharmacophoric template for identifying and designing small-molecule inhibitors of hHsp90 dimerization. PMID:24760083

  7. Unusual nanosized associates of carboxy-calix[4]resorcinarene and cetylpyridinium chloride: the macrocycle as a glue for surfactant micelles.

    Science.gov (United States)

    Morozova, Ju E; Syakaev, V V; Shalaeva, Ya V; Ermakova, A M; Nizameev, I R; Kadirov, M K; Voloshina, A D; Zobov, V V; Antipin, I S; Konovalov, A I

    2017-03-08

    The association of cetylpyridinium chloride (CPC) micelles in the presence of octaacetated tetraphenyleneoxymethylcalix[4]resorcinarene (CR) leads to the formation of unusual spherical supramolecular nanoparticles (SNPs). Within the range of CR/CPC molar ratios from 10/1 to 1/10 (except for 1/8), CR, acting as a counterion, decreases the critical micelle concentration of CPC by one order of magnitude and leads to the formation of SNPs with an average hydrodynamic radius of 164 nm and an average zeta potential of -60 mV. The formation of SNPs was studied by NMR FT-PGSE and 2D NOESY, DLS, TEM, fluorimetry, and UV-Vis methods. The stability of SNPs at different temperatures and pH values and in the presence of electrolytes was investigated. The specificity of the interactions of the SNPs with substrates that were preferentially bound by a macrocycle or CPC micelle was studied. The enhancement of cation dye binding in the presence of SNPs is shown. The presented supramolecular system may serve as a nanocapsule for water-soluble and water-insoluble compounds.

  8. Synthesis and characterization of novel 2, 2'-bipyrimidine fluorescent derivative for protein binding

    Directory of Open Access Journals (Sweden)

    Padalkar Vikas S

    2011-11-01

    Full Text Available Abstract Background Fluorescent dyes with biocompatible functional group and good fluorescence behavior are used as biosensor for monitoring different biological processes as well as detection of protein assay. All reported fluorophore used as sensors are having high selectivity and sensitivity but till there is more demand to synthesized new fluorophore which have improved fluorescence properties and good biocompatibility. Results Novel 4, 4'-(1, 1'-(5-(2-methoxyphenoxy-[2, 2'-bipyrimidine]-4, 6-diylbis(1H-pyrazol-3, 1-diyl dianiline fluorescent dye was synthesized by multistep synthesis from 2-phenylacetonitrile, 2-chloropyrimidine and 2-methoxyphenol. This dye has absorption at 379 nm with intense single emission at 497 nm having fairly good quantum yield (0.375 and Stokes shift. The intermediates and dye were characterized by FT-IR, 1H NMR, 13C NMR and Mass spectral analysis. The pyrazole bipyrimidine based fluorescent dye possessing two amino groups suitable for binding with protein is reported. Its utility as a biocompatible conjugate was explained by conjugation with bovine serum albumin. The method is based on direct fluorescence detection of fluorophore-labelled protein before and after conjugation. Purified fluorescent conjugate was subsequently analyzed by fluorimetry. The analysis showed that the tested conjugation reaction yielded fluorescent conjugates of the dye through carbodiimide chemistry. Conclusion In summery synthesized fluorophore pyrazole-bipyrimidine has very good interaction towards protein bovine serum albumin and it acts as good candidate for protein assay.

  9. Use of Magnetic Folate-Dextran-Retinoic Acid Micelles for Dual Targeting of Doxorubicin in Breast Cancer

    Directory of Open Access Journals (Sweden)

    J. Varshosaz

    2013-01-01

    Full Text Available Amphiphilic copolymer of folate-conjugated dextran/retinoic acid (FA/DEX-RA was self-assembled into micelles by direct dissolution method. Magnetic iron oxide nanoparticles (MNPs coated with oleic acid (OA were prepared by hydrothermal method and encapsulated within the micelles. Doxorubicin HCl was loaded in the magnetic micelles. The characteristics of the magnetic micelles were determined by Fourier transform infrared (FT-IR spectroscopy, thermogravimetric analysis (TGA, transmission electron microscopy (TEM, and vibrating sample magnetometer (VSM. The crystalline state of OA-coated MNPs and their heat capacity were analyzed by X-ray diffraction (XRD and differential scanning calorimetry (DSC methods, respectively. The iron content of magnetic micelles was determined using inductively coupled plasma optical emission spectrometry (ICP-OES. Bovine serum albumin (BSA was used to test the protein binding of magnetic micelles. The cytotoxicity of doxorubicin loaded magnetic micelles was studied on MCF-7 and MDA-MB-468 cells using MTT assay and their quantitative cellular uptake by fluorimetry method. TEM results showed the MNPs in the hydrophobic core of the micelles. TGA results confirmed the presence of OA and FA/DEX-RA copolymer on the surface of MNPs and micelles, respectively. The magnetic micelles showed no significant protein bonding and reduced the IC50 of the drug to about 10 times lower than the free drug.

  10. Fluorinated ionic liquids for protein drug delivery systems: Investigating their impact on the structure and function of lysozyme.

    Science.gov (United States)

    Alves, Márcia; Vieira, Nicole S M; Rebelo, Luís Paulo N; Araújo, João M M; Pereiro, Ana B; Archer, Margarida

    2017-06-30

    Since the approval of recombinant human insulin by FDA in 1982, more than 200 proteins are currently available for pharmaceutical use to treat a wide range of diseases. However, innovation is still required to develop effective approaches for drug delivery. Our aim is to investigate the potential use of fluorinated ionic liquids (FILs) as drug delivery systems (DDS) for therapeutic proteins. Some initial parameters need to be assessed before further studies can proceed. This work evaluates the impact of FILs on the stability, function, structure and aggregation state of lysozyme. Different techniques were used for this purpose, which included differential scanning fluorimetry (DSF), spectrophotometric assays, circular dichroism (CD), dynamic light scattering (DLS), and scanning and transmission electron microscopy (SEM/TEM). Ionic liquids composed of cholinium-, imidazolium- or pyridinium- derivatives were combined with different anions and analysed at different concentrations in aqueous solutions (below and above the critical aggregation concentration, CAC). The results herein presented show that the addition of ionic liquids had no significant effect on the stability and hydrolytic activity of lysozyme. Moreover, a distinct behaviour was observed in DLS experiments for non-surfactant and surfactant ionic liquids, with the latter encapsulating the protein at concentrations above the CAC. These results encourage us to further study ionic liquids as promising tools for DDS of protein drugs. Copyright © 2017 Elsevier B.V. All rights reserved.

  11. Delineating Substrate Diversity of Disparate Short-Chain Dehydrogenase Reductase from Debaryomyces hansenii.

    Directory of Open Access Journals (Sweden)

    Arindam Ghatak

    Full Text Available Short-chain dehydrogenase reductases (SDRs have been utilized for catalyzing the reduction of many aromatic/aliphatic prochiral ketones to their respective alcohols. However, there is a paucity of data that elucidates their innate biological role and diverse substrate space. In this study, we executed an in-depth biochemical characterization and substrate space mapping (with 278 prochiral ketones of an unannotated SDR (DHK from Debaryomyces hansenii and compared it with structurally and functionally characterized SDR Synechococcus elongatus. PCC 7942 FabG to delineate its industrial significance. It was observed that DHK was significantly more efficient than FabG, reducing a diverse set of ketones albeit at higher conversion rates. Comparison of the FabG structure with a homology model of DHK and a docking of substrate to both structures revealed the presence of additional flexible loops near the substrate binding site of DHK. The comparative elasticity of the cofactor and substrate binding site of FabG and DHK was experimentally substantiated using differential scanning fluorimetry. It is postulated that the loop flexibility may account for the superior catalytic efficiency of DHK although the positioning of the catalytic triad is conserved.

  12. Spectroscopic Analysis of Red Fluorescent Proteins and Development of a Microfluidic Cell Sorter for the Generation of Improved Variants

    Science.gov (United States)

    Lubbeck, Jennifer L.

    The discovery of the green fluorescent protein (GFP) launched the development of a wide variety of fluorescent protein (FP) mutants whose spectral and photophysical diversity revolutionized in vivo imaging. The excitation and emission spectra of red fluorescent proteins (RFPs), in particular, have been ideally tuned to a window optically favorable for in vivo work. However, their quantum yields, photostabilities and fluorescence intermittency properties require improvement if they are to be broadly employed for low-copy or single-molecule measurements. Attempts to engineer improved RFPs often result in optimization of one photophysical property at the expense of others. We developed a microfluidic-based cytometer for screening HeLa cell-based genetic RFP-libraries simultaneously on the basis of fluorescence lifetime (a proxy for quantum yield), photostability, and brightness. Ten 532 nm excitation beams interrogate each cell in flow. The first is electro-optically modulated (30 MHz) to enable lifetime measurement with phase fluorimetry. The remaining beams act as a pulse sequence for isolating the irreversible photobleaching time constant. Optical-force switching is employed to sort cells based on any combination of the photophysical parameters. Screening with this instrument enables identification of regions of the structure that synergistically affect quantum yield and photostability and the sorting capability provides a new tool for accelerating the development of next generation RFPs.

  13. Synergistic effect of MiR-146a mimic and cetuximab on hepatocellular carcinoma cells.

    Science.gov (United States)

    Huang, Suning; He, Rongquan; Rong, Minhua; Dang, Yiwu; Chen, Gang

    2014-01-01

    Previously, we found that the expression of microRNA-146a (miR-146a) was downregulated in hepatocellular carcinoma (HCC) formalin-fixed paraffin-embedded (FFPE) tissues compared to the adjacent noncancerous hepatic tissues. In the current study, we have explored the in vitro effect of miR-146a on the malignant phenotypes of HCC cells. MiR-146a mimic could suppress cell growth and increase cellular apoptosis in HCC cell lines HepG2, HepB3, and SNU449, as assessed by spectrophotometry, fluorimetry, and fluorescence microscopy, respectively. Furthermore, western blot showed that miR-146a mimic downregulated EGFR, ERK1/2, and stat5 signalings. These effects were less potent compared to that of a siRNA targeting EGFR, a known target gene of miR-146a. Moreover, miR-146a mimic could enhance the cell growth inhibition and apoptosis induction impact of various EGFR targeting agents. The most potent combination was miR-146a mimic with cetuximab, presenting a synergistic effect. In conclusion, miR-146a plays a vital role in the cell growth and apoptosis of HCC cells and inducing miR-146a level might be a critical targeted molecular therapy strategy for HCC.

  14. Role of chemistry in uranium leaching from sand stone ore bodies by ISL technique (abstract)

    International Nuclear Information System (INIS)

    Sheikh, A.K.; Hanif, M.

    2011-01-01

    The importance of uranium for nuclear power generation is increasing day by day in the current energy requirement scenario. The energy sector is reverting to nuclear power generation in many countries. Keeping in view the limited availability of fossil fuels and the problems associated with its exploration. In situ leach mining of uranium is currently employed in many countries for extraction of uranium in soft rocks from subsurface where deposits are lying deeper and the cost of underground mining (UGM) are high. In situ leaching of uranium revolves around injection of leachant (usually bicarbonate) and oxidant into the host rock without disturbing the strata thereby solubilizing the uranium in solution form. The resultant leach is then pumped out and sent to processing plant for production of yellow cake. The Chemistry Division NMC-I, D. G. Khan plays a vital role in ISL Mining of uranium, starting from initial exploration of ore body, analysis of leach liquor and other liquid samples up to the final step of yellow cake formation. During all these analysis to ensure Quality control, Chemistry Division utilizes modern / updated analytical techniques e.g. Atomic Absorption Spectroscopy, Spectrophotometry, Fluorimetry and Potentiometry. R and D studies are also being carried out to optimize and economize the operational parameters accordingly. This presentation basically encompasses the role of Chemistry at various levels and stages at Mining Projects to achieve this goal. (author)

  15. Antioxidant effects of herbal therapies used by patients with inflammatory bowel disease: an in vitro study.

    Science.gov (United States)

    Langmead, L; Dawson, C; Hawkins, C; Banna, N; Loo, S; Rampton, D S

    2002-02-01

    Herbal remedies used by patients for treatment of inflammatory bowel disease include slippery elm, fenugreek, devil's claw, Mexican yam, tormentil and wei tong ning, a traditional Chinese medicine. Reactive oxygen metabolites produced by inflamed colonic mucosa may be pathogenic. Aminosalicylates (5-ASA) are antioxidant and other such agents could be therapeutic. To assess the antioxidant effects of herbal remedies in cell-free oxidant-generating systems and inflamed human colorectal biopsies. Luminol-enhanced chemiluminescence in a xanthine/xanthine oxidase cell-free system was used to detect superoxide scavenging by herbs and 5-ASA, and fluorimetry to define peroxyl radical scavenging using a phycoerythrin degradation assay. Chemiluminescence was used to detect herbal effects on generation of oxygen radicals by mucosal biopsies from patients with active ulcerative colitis. Like 5-ASA, all herbs, except fenugreek, scavenged superoxide dose-dependently. All materials tested scavenged peroxyl dose-dependently. Oxygen radical release from biopsies was reduced after incubation in all herbs except Mexican yam, and by 5-ASA. All six herbal remedies have antioxidant effects. Fenugreek is not a superoxide scavenger, while Mexican yam did not inhibit radical generation by inflamed biopsies. Slippery elm, fenugreek, devil's claw, tormentil and wei tong ning merit formal evaluation as novel therapies in inflammatory bowel disease.

  16. Uranium concentration in building materials used in the central region of Egypt

    International Nuclear Information System (INIS)

    Higgy, R.H.; El-Tahawy, M.S.; Ghods, A.

    1997-01-01

    Within a radiological survey of the building materials used in the urban dwellings in the central region of Egypt, the uranium concentration in 80 representative samples of raw and fabricated building materials are determined using laser fluorimetry technique. For 40 samples from the studied raw building materials of sand, gravel, gypsum, lime-stone, granite and marble the determined uranium concentration values range between 0.3 and 3.6 ppm for all these samples except for one type of granite having the corresponding value of 7.8 ppm. For 37 samples from studied fabricated building materials of normal cement, clay brick, sand brick, tiles and ceramic plates the determined uranium concentration values range from 0.5 to 3.4 ppm. The corresponding values for three types of iron cement are 3.1, 6.1 and 9.3 ppm. The radium-226 content (of the uranium-238 series) in the same samples was determined using high resolution gamma-ray spectrometers based on HP Ge-detectors. The data obtained by the two techniques are in good agreement for the majority of the studied samples. (author)

  17. Report of radioactivity survey in Kanagawa Prefecture, 2000

    International Nuclear Information System (INIS)

    2001-03-01

    This report involves the document of investigations conducted in the prefecture from January 1 to December 31, 2000, and the actual data of general levels of food and environment radiation and radioactivity and of the uranium levels around the nuclear fuel processing facility (Japan Nuclear Fuel Ltd.). Investigations included were: the nuclide analysis by γ-ray spectrometry using Ge-SSD multi-channel analyzer of rain, land and sea waters, fall-out, river and sea sediments, soil and foods; the estimation of uranium levels by solid fluorimetry in river and sea waters, soil, river and sea sediments and sea weeds; the measurement of gross beta-ray with GM-counters; the ambient dose rate with NaI scintillation survey meter and so on. Radon measurement and on-site survey at call of nuclear powered ships in Yokosuka US Navy Base were also performed. Data on environment and food showed no change as compared with those in the previous year. Uranium levels were normal. Foods brought into the prefecture from Japan Sea showed the similar radioactivity levels to those from Sagami Bay. Call of the ships at the Base increased to 135% in comparison with that in the previous year. (K.H.)

  18. Application of a high-throughput relative chemical stability assay to screen therapeutic protein formulations by assessment of conformational stability and correlation to aggregation propensity.

    Science.gov (United States)

    Rizzo, Joseph M; Shi, Shuai; Li, Yunsong; Semple, Andrew; Esposito, Jessica J; Yu, Shenjiang; Richardson, Daisy; Antochshuk, Valentyn; Shameem, Mohammed

    2015-05-01

    In this study, an automated high-throughput relative chemical stability (RCS) assay was developed in which various therapeutic proteins were assessed to determine stability based on the resistance to denaturation post introduction to a chaotrope titration. Detection mechanisms of both intrinsic fluorescence and near UV circular dichroism (near-UV CD) are demonstrated. Assay robustness was investigated by comparing multiple independent assays and achieving r(2) values >0.95 for curve overlays. The complete reversibility of the assay was demonstrated by intrinsic fluorescence, near-UV CD, and biologic potency. To highlight the method utility, we compared the RCS assay with differential scanning calorimetry and dynamic scanning fluorimetry methodologies. Utilizing C1/2 values obtained from the RCS assay, formulation rank-ordering of 12 different mAb formulations was performed. The prediction of long-term stability on protein aggregation is obtained by demonstrating a good correlation with an r(2) of 0.83 between RCS and empirical aggregation propensity data. RCS promises to be an extremely useful tool to aid in candidate formulation development efforts based on the complete reversibility of the method to allow for multiple assessments without protein loss and the strong correlation between the C1/2 data obtained and accelerated stability under stressed conditions. © 2015 Wiley Periodicals, Inc. and the American Pharmacists Association.

  19. Divers models of divalent cation interaction to calcium-binding proteins: techniques and anthology.

    Science.gov (United States)

    Cox, Jos A

    2013-01-01

    Intracellular Ca(2+)-binding proteins (CaBPs) are sensors of the calcium signal and several of them even shape the signal. Most of them are equipped with at least two EF-hand motifs designed to bind Ca(2+). Their affinities are very variable, can display cooperative effects, and can be modulated by physiological Mg(2+) concentrations. These binding phenomena are monitored by four major techniques: equilibrium dialysis, fluorimetry with fluorescent Ca(2+) indicators, flow dialysis, and isothermal titration calorimetry. In the last quarter of the twentieth century reports on the ion-binding characteristics of several abundant wild-type CaBPs were published. With the advent of recombinant CaBPs it became possible to determine these properties on previously inaccessible proteins. Here I report on studies by our group carried out in the last decade on eight families of recombinant CaBPs, their mutants, or truncated domains. Moreover this chapter deals with the currently used methods for quantifying the binding of Ca(2+) and Mg(2+) to CaBPs.

  20. Depolarization of the conductance-voltage relationship in the NaV1.5 mutant, E1784K, is due to altered fast inactivation.

    Directory of Open Access Journals (Sweden)

    Colin H Peters

    Full Text Available E1784K is the most common mixed long QT syndrome/Brugada syndrome mutant in the cardiac voltage-gated sodium channel NaV1.5. E1784K shifts the midpoint of the channel conductance-voltage relationship to more depolarized membrane potentials and accelerates the rate of channel fast inactivation. The depolarizing shift in the midpoint of the conductance curve in E1784K is exacerbated by low extracellular pH. We tested whether the E1784K mutant shifts the channel conductance curve to more depolarized membrane potentials by affecting the channel voltage-sensors. We measured ionic currents and gating currents at pH 7.4 and pH 6.0 in Xenopus laevis oocytes. Contrary to our expectation, the movement of gating charges is shifted to more hyperpolarized membrane potentials by E1784K. Voltage-clamp fluorimetry experiments show that this gating charge shift is due to the movement of the DIVS4 voltage-sensor being shifted to more hyperpolarized membrane potentials. Using a model and experiments on fast inactivation-deficient channels, we show that changes to the rate and voltage-dependence of fast inactivation are sufficient to shift the conductance curve in E1784K. Our results localize the effects of E1784K to DIVS4, and provide novel insight into the role of the DIV-VSD in regulating the voltage-dependencies of activation and fast inactivation.

  1. A structural basis for cellular uptake of GST-fold proteins.

    Directory of Open Access Journals (Sweden)

    Melanie J Morris

    Full Text Available It has recently emerged that glutathione transferase enzymes (GSTs and other structurally related molecules can be translocated from the external medium into many different cell types. In this study we aim to explore in detail, the structural features that govern cell translocation and by dissecting the human GST enzyme GSTM2-2 we quantatively demonstrate that the α-helical C-terminal domain (GST-C is responsible for this property. Attempts to further examine the constituent helices within GST-C resulted in a reduction in cell translocation efficiency, indicating that the intrinsic GST-C domain structure is necessary for maximal cell translocation capacity. In particular, it was noted that the α-6 helix of GST-C plays a stabilising role in the fold of this domain. By destabilising the conformation of GST-C, an increase in cell translocation efficiency of up to ∼2-fold was observed. The structural stability profiles of these protein constructs have been investigated by circular dichroism and differential scanning fluorimetry measurements and found to impact upon their cell translocation efficiency. These experiments suggest that the globular, helical domain in the 'GST-fold' structural motif plays a role in influencing cellular uptake, and that changes that affect the conformational stability of GST-C can significantly influence cell translocation efficiency.

  2. The seleno-organic compound ebselen impairs mitochondrial physiology and induces cell death in AR42J cells.

    Science.gov (United States)

    Santofimia-Castaño, Patricia; Garcia-Sanchez, Lourdes; Ruy, Deborah Clea; Fernandez-Bermejo, Miguel; Salido, Gines M; Gonzalez, Antonio

    2014-09-17

    Ebselen is a seleno-organic compound that causes cell death in several cancer cell types. The mechanisms underlying its deleterious effects have not been fully elucidated. In this study, the effects of ebselen (1 μM-40 μM) on AR42J tumor cells have been examined. Cell viability was studied using AlamarBlue(®) test. Cell cycle phase determination was carried out by flow cytometry. Changes in intracellular free Ca(2+) concentration were followed by fluorimetry analysis of fura-2-loaded cells. Distribution of mitochondria, mitochondrial Ca(2+) concentration and mitochondrial membrane potential were monitored by confocal microscopy of cells loaded with Mitotracker Green™ FM, rhod-2 or TMRM respectively. Caspase-3 activity was calculated following the luorogenic substrate ACDEVD-AMC signal with a spectrofluorimeter. Results show that cell viability decreased in the presence of ebselen. An increase in the number of cells in the S-phase of the cell cycle was observed. Ebselen induced a concentration-dependent mobilization of Ca(2+) from agonist- and thapsigargin-sensitive Ca(2+) pools. Ebselen induced also a transient increase in mitochondrial Ca(2+) concentration, a progressive decrease of the mitochondrial membrane potential and a disruption of the mitochondrial network. Finally, a concentration-dependent increase in caspase-3 activity was detected. We conclude that ebselen exerts deleterious actions on the cells that involve the impairment of mitochondrial physiology and the activation of caspase-3-mediated apoptotic pathway. Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.

  3. Smectite clays of Serbia and their application in adsorption of organic dyes

    Science.gov (United States)

    Milošević, Maja; Logar, Mihovil

    2014-05-01

    Colorants and dyes are currently available in over a 100.000 different species and several biggest industries are using them daily in their manufacture processes (textile, cosmetics, food industry, etc.). Since colorants are easily dissoluble in water they pass through filter membranes without further decomposing and in that manner they end up in the environment. The main goal of this work is to apply certain methods in determining the suitability of individual clay in adsorbing and removing colorants from polluted waters. For this study we have chosen four different raw clays from three regions in Serbia: Svrljig (B), Bogovina (Bo) and Slatina-Ub (C and V) and as colorant - methylene blue dye (MB (MERCK, for analytical purposes)). Experiments where carried out to determine the sample structure (XRD and IR), grain size (granulometry), cationic exchange capacity (CEC via spectrophotometry using MB) and adsorption capabilities (spectrophotometry and fluorimetry using MB). XRD and IR data are showing that the samples are smectite clays where samples B i Bo are mainly montmorillonite while C and V are montmorillonite-illite clays. Granulometric distribution results indicate that samples B i Bo have smaller grain size, less that 1μ (over 60%) whereas the samples C and V are more coarse grained (40% over 20μ). This grain distribution is affecting their specific surface area in the manner that those coarse grained samples have smaller specific surface area. Cationic exchange capacity determined with methylene blue indicate that montmorillonite samples have larger CEC (B = 37 meq/100g, Bo = 50 meq/100g) and montmorillonite-illite samples smaller CEC (V = 5 meq/100g, V = 3 meq/100g). Fluorimetry measurement results gave us a clear distinction between those with higher and smaller adsorption capability. Montmorillonite samples (B and Bo) with higher CEC values and smaller grain size are adsorbing large amounts of methylene blue witch is visible by absence of fluorimetric

  4. Contributions to anionexchange separation of metal ions in mixed aqueous-organic solvent systems

    International Nuclear Information System (INIS)

    Cunha-Campos, C.

    1986-01-01

    In the first chapter of the thesis a method is described for the determination of iron, copper, manganese, nickel, cobalt and uranium in samples of manganese nodules. After dissolution of the sample in a mixture of concentrated hydrochloric-nitric acids, the elements are adsorbed on a column of the strongly basic anion-exchange resin Dowex 1 from a medium consisting of 80% cyclohexanone 10% methanol and 10% 1M hydrochloric acid. Following removal of iron by washing the resin bed with a mixture of the same composition, 90% methanol-10% 1M hydrochloric containing ascorbic acid is passed through the column to eluate manganese, nickel and cobalt. Subsequently, 4M hydrochloric acid-1% hydrogen peroxid solution and 1M hydrochloric acid are used to elute copper and uranium, respectively. The elements were determined quantitatively by ICP-OES, except for uranium which was determined by fluorimetry. The method was used successfully for the determination of mg and p.p.m. quantities of these elements in samples of manganese nodules. In the second chapter of the thesis a procedure is described for the determination of cadmium, lead, zinc and uranium in samples of Austrian mineral waters. After acidification, the sample was heated to expell CO 2 and the elements in the presence of 8-hydroxyquinoline are extracted into chloroform at pH-7. Then the metals are adsorbed on a column of the strongly basic anion-exchange resin Dowex 1 from a medium consisting of 55% chloroform, 36% methanol and 9% 12M hydrochloric acid. After simultaneous elution of the elements with 1M nitric acid, a portion of the eluate was passed through another column containing the same resin to separate uranium from the other elements. The uranium was determined fluorimetrically and cadmium, lead and zinc by atomic absorption spectroscopy. The method was successfully used for the quantitative determination of these elements in mineral water samples. (Author)

  5. A novel cross-species inhibitor to study the function of CatSper Ca2+ channels in sperm.

    Science.gov (United States)

    Rennhack, Andreas; Schiffer, Christian; Brenker, Christoph; Fridman, Dmitry; Nitao, Elis T; Cheng, Yi-Min; Tamburrino, Lara; Balbach, Melanie; Stölting, Gabriel; Berger, Thomas K; Kierzek, Michelina; Alvarez, Luis; Wachten, Dagmar; Zeng, Xu-Hui; Baldi, Elisabetta; Publicover, Stephen; Kaupp, U Benjamin; Strünker, Timo

    2018-05-03

    Sperm from many species share the sperm-specific Ca 2+ channel CatSper (cation channel of sperm) that controls the intracellular Ca 2+ concentration and, thereby, the swimming behaviour. A growing body of evidence suggests that the mechanisms controlling CatSper activity and the role of the channel during fertilization differ among species. However, a lack of suitable pharmacological tools has hampered the elucidation of the function of CatSper. Known CatSper inhibitors exhibit considerable side effects and inhibit also Slo3, the K + channel in mammalian sperm. The drug RU1968 was reported to suppress Ca 2+ signaling in human sperm by an unknown mechanism. We resynthesized the drug and revisited its mechanism of action in sperm form humans, mice, and sea urchins. We show by Ca 2+ fluorimetry, single-cell Ca 2+ imaging, electrophysiology, opto-chemistry, and motility analysis that RU1968 inhibits CatSper in sperm from invertebrates and mammals. The drug lacks toxic side effects in human sperm, does not affect mouse Slo3, and inhibits human Slo3 with about 15-fold lower potency than CatSper. Moreover, in human sperm, the inhibitor mimics CatSper dysfunction and suppresses motility responses evoked by progesterone, an oviductal steroid that activates CatSper. Finally, we show that the drug abolishes CatSper-mediated chemotactic navigation in sea urchin sperm. We propose RU1968 as a novel tool to elucidate the function of CatSper in sperm across species. This article is protected by copyright. All rights reserved.

  6. In situ determination of the depuration of three- and four-ringed polycyclic aromatic hydrocarbons co-adsorbed onto mangrove leaf surfaces

    International Nuclear Information System (INIS)

    Sun, Haifeng; Shi, Jing; Guo, Shuai; Zhang, Yong; Duan, Lusha

    2016-01-01

    A dual-wavelength fiber-optic fluorimetry for the in situ simultaneous determinations of fluorene (Flu), phenanthrene (Phe) and pyrene (Pyr) adsorbed onto the leaf surfaces of living Avicennia marina (Am) seedling were developed and used to study the depuration kinetics of the three PAHs, adsorbed individually or mixed together, onto living Am leaf surfaces. Limits of detection for the in situ measurements of adsorbed Flu, Phe and Pyr were 4.62, 2.75 and 1.38 ng spot"−"1, respectively. The depuration kinetics of the three selected polycyclic aromatic hydrocarbons (PAHs) are divided into rapid and slow phases; both phases followed the same first-order kinetics with relative clearance rates of Flu > Phe > Pyr during the rapid phase, and a clearance rate order of Pyr > Flu > Phe during the slow phase. For the three PAHs co-adsorbed on living Am leaf surfaces, a significant synergistic effect was detected during the rapid phase clearance; conversely, an antagonistic effect was observed during the slow phase. However, the synergistic effect dominated during both phases of the depuration process, and the co-adsorption of PAHs promoted the clearance of all three compounds from the mangrove leaf surfaces. These findings demonstrate a novel analytical method for in situ characterization of multiple PAHs adsorbed onto the plant surfaces. - Highlights: • A novel method for the in situ determination of multi-component PAHs was developed. • Synergistic and antagonistic effects separately occurred over rapid and slow phases. • The clearance of all three PAHs from leaf surfaces was promoted by co-adsorption. - The co-adsorption of PAHs promoted the depuration of all three compounds from the mangrove leaf surfaces.

  7. Determination of blood cyanide by HPLC-MS.

    Science.gov (United States)

    Tracqui, A; Raul, J S; Géraut, A; Berthelon, L; Ludes, B

    2002-04-01

    An original high-performance liquid chromatographic-mass spectrometric (HPLC-MS) procedure was developed for the determination of cyanide (CN) in whole blood. After the addition of K13C15N as internal standard, blood was placed in a microdiffusion device, the inner well of which was filled with a mixture of taurine (50mM in water)/naphthalene-2,3-dicarboxaldehyde (NDA, 10mM in methanol)/methanol/ concentrated (approximately 20%) ammonia solution (25:25:45:5, v/v). Concentrated H2SO4 was added to the blood sample, and the microdiffusion chamber was sealed. After 30 min of gentle agitation, 2 microL of the contents of the inner vial were pipetted and directly injected onto a NovaPak C18 HPLC column. Separation was performed by a gradient of acetonitrile in 2mM NH4COOH, pH 3.0 buffer (35-80% in 10 min). Detection was done with a Perkin-Elmer Sciex API-100 mass analyzer with an ionspray interface, operated in the negative ionization mode. MS data were collected as either TIC or SIM at m/z (299 + 191) and (301 + 193) for the derivatives formed with CN and 13C15N, respectively. Inspired by previous works dealing with the complexation of CN by NDA + taurine to form a 1-cyano [f] benzoisoindole derivative analyzed by HPLC-fluorimetry, this method appears simple, rapid, and extremely specific. Limits of detection and quantitation for blood CN are 5 and 15 ng/mL, respectively. The use of 13C15N as internal standard allows the quantitation of CN with elegance and accuracy in comparison with previously reported methods.

  8. Thymosin beta 4 protects cardiomyocytes from oxidative stress by targeting anti-oxidative enzymes and anti-apoptotic genes.

    Directory of Open Access Journals (Sweden)

    Chuanyu Wei

    Full Text Available Thymosin beta-4 (Tβ4 is a ubiquitous protein with many properties relating to cell proliferation and differentiation that promotes wound healing and modulates inflammatory mediators. The mechanism by which Tβ4 modulates cardiac protection under oxidative stress is not known. The purpose of this study is to dissect the cardioprotective mechanism of Tβ4 on H(2O(2 induced cardiac damage.Rat neonatal cardiomyocytes with or without Tβ4 pretreatment were exposed to H(2O(2 and expression of antioxidant, apoptotic, and anti-inflammatory genes was evaluated by quantitative real-time PCR and western blotting. ROS levels were estimated by DCF-DA using fluorescent microscopy and fluorimetry. Selected antioxidant, anti-inflammatory and antiapoptotic genes were silenced by siRNA transfections in neonatal cardiomyocytes and effect of Tβ4 on H(2O(2-induced cardiac damage was evaluated.Pre-treatment of Tβ4 resulted in reduction of the intracellular ROS levels induced by H(2O(2 in cardiomyocytes. Tβ4 pretreatment also resulted in an increase in the expression of antiapoptotic proteins and reduction of Bax/BCl(2 ratio in the cardiomyocytes. Pretreatment with Tβ4 resulted in stimulating the expression of antioxidant enzymes copper/zinc SOD and catalase in cardiomyocytes at both transcription and translation levels. Tβ4 treatment resulted in the increased expression of anti-apoptotic and anti-inflammatory genes. Silencing of Cu/Zn SOD and catalase gene resulted in apoptotic cell death in the cardiomyocytes which was prevented by treatment with Tβ4.This is the first report that demonstrates the effect of Tβ4 on cardiomyocytes and its capability to selectively upregulate anti-oxidative enzymes, anti-inflammatory genes, and antiapoptotic enzymes in the neonatal cardiomyocytes thus preventing cell death thereby protecting the myocardium. Tβ4 treatment resulted in decreased oxidative stress and inflammation in the myocardium under oxidative stress.

  9. Vitamin K3 (menadione)-induced oncosis associated with keratin 8 phosphorylation and histone H3 arylation.

    Science.gov (United States)

    Scott, Gary K; Atsriku, Christian; Kaminker, Patrick; Held, Jason; Gibson, Brad; Baldwin, Michael A; Benz, Christopher C

    2005-09-01

    The vitamin K analog menadione (K3), capable of both redox cycling and arylating nucleophilic substrates by Michael addition, has been extensively studied as a model stress-inducing quinone in both cell culture and animal model systems. Exposure of keratin 8 (k-8) expressing human breast cancer cells (MCF7, T47D, SKBr3) to K3 (50-100 microM) induced rapid, sustained, and site-specific k-8 serine phosphorylation (pSer73) dependent on signaling by a single mitogen activated protein kinase (MAPK) pathway, MEK1/2. Normal nuclear morphology and k-8 immunofluorescence coupled with the lack of DNA laddering or other features of apoptosis indicated that K3-induced cytotoxicity, evident within 4 h of treatment and delayed but not prevented by MEK1/2 inhibition, was due to a form of stress-activated cell death known as oncosis. Independent of MAPK signaling was the progressive appearance of K3-induced cellular fluorescence, principally nuclear in origin and suggested by in vitro fluorimetry to have been caused by K3 thiol arylation. Imaging by UV transillumination of protein gels containing nuclear extracts from K3-treated cells revealed a prominent 17-kDa band shown to be histone H3 by immunoblotting and mass spectrometry (MS). K3 arylation of histones in vitro followed by electrospray ionization-tandem MS analyses identified the unique Cys110 residue within H3, exposed only in the open chromatin of transcriptionally active genes, as a K3 arylation target. These findings delineate new pathways associated with K3-induced stress and suggest a potentially novel role for H3 Cys110 as a nuclear stress sensor.

  10. Chimeric β-Lactamases: Global Conservation of Parental Function and Fast Time-Scale Dynamics with Increased Slow Motions

    Science.gov (United States)

    Clouthier, Christopher M.; Morin, Sébastien; Gobeil, Sophie M. C.; Doucet, Nicolas; Blanchet, Jonathan; Nguyen, Elisabeth; Gagné, Stéphane M.; Pelletier, Joelle N.

    2012-01-01

    Enzyme engineering has been facilitated by recombination of close homologues, followed by functional screening. In one such effort, chimeras of two class-A β-lactamases – TEM-1 and PSE-4 – were created according to structure-guided protein recombination and selected for their capacity to promote bacterial proliferation in the presence of ampicillin (Voigt et al., Nat. Struct. Biol. 2002 9:553). To provide a more detailed assessment of the effects of protein recombination on the structure and function of the resulting chimeric enzymes, we characterized a series of functional TEM-1/PSE-4 chimeras possessing between 17 and 92 substitutions relative to TEM-1 β-lactamase. Circular dichroism and thermal scanning fluorimetry revealed that the chimeras were generally well folded. Despite harbouring important sequence variation relative to either of the two ‘parental’ β-lactamases, the chimeric β-lactamases displayed substrate recognition spectra and reactivity similar to their most closely-related parent. To gain further insight into the changes induced by chimerization, the chimera with 17 substitutions was investigated by NMR spin relaxation. While high order was conserved on the ps-ns timescale, a hallmark of class A β-lactamases, evidence of additional slow motions on the µs-ms timescale was extracted from model-free calculations. This is consistent with the greater number of resonances that could not be assigned in this chimera relative to the parental β-lactamases, and is consistent with this well-folded and functional chimeric β-lactamase displaying increased slow time-scale motions. PMID:23284969

  11. Uranium and other heavy toxic elements distribution in the drinking water samples of SW-Punjab, India - a potentially dangerous trend

    International Nuclear Information System (INIS)

    Bajwa, B.S.

    2015-01-01

    In the present investigations, Laser Fluorimetry technique has been used for the microanalysis of uranium content in drinking water samples collected from different sources like the hand pumps, tube wells of various depths from wide range of locations in the four districts of SW-Punjab, India. The purpose of this study was to investigate the uranium concentration levels of ground water being used for drinking purposes and to determine its health effects, if any, to the local population of this region. Corresponding radiological and chemical risks have also been calculated for the uranium concentrations in ground water samples. Some other heavy elements have also been analysed using the Atomic Absorption Spectrometry. In this region, uranium concentration in 498 drinking water samples has been found to vary between 0.5-579 μgl -1 with an average of 73.5 μgl -1 , Data analysis revealed that 338 of 498 samples had uranium concentration higher than recommended safe limit of 30 μgl -1 (WHO, 2011) while 216 samples exceeded the threshold of 60 μgl -1 recommended by AERB, DAE, India, 2004. Overall data analysis reveals that, 68% of the collected samples have uranium concentration higher than safe limit of 30 μgl -1 (WHO, 2011) while 43% samples exceed the threshold of 60μgl -1 recommended by AERB, DAE, India, 2004. Higher concentrations observed in ground water samples of SW-Punjab might be due to leaching of uranium from adjoining/basement granite rich rock formations. The anthropogenic activities, urbanization and wide spread use of pesticides/fertilizers which is responsible for increase in the TDS/salinity of the region might be another cause. But, overall it seems that the plausible source of high uranium observed in this region may be of geogenic in nature. (author)

  12. Expression and activation of the oxytocin receptor in airway smooth muscle cells: Regulation by TNFα and IL-13

    Directory of Open Access Journals (Sweden)

    Siddiqui Salman

    2010-07-01

    Full Text Available Abstract Background During pregnancy asthma may remain stable, improve or worsen. The factors underlying the deleterious effect of pregnancy on asthma remain unknown. Oxytocin is a neurohypophyseal protein that regulates a number of central and peripheral responses such as uterine contractions and milk ejection. Additional evidence suggests that oxytocin regulates inflammatory processes in other tissues given the ubiquitous expression of the oxytocin receptor. The purpose of this study was to define the role of oxytocin in modulating human airway smooth muscle (HASMCs function in the presence and absence of IL-13 and TNFα, cytokines known to be important in asthma. Method Expression of oxytocin receptor in cultured HASMCs was performed by real time PCR and flow cytomery assays. Responses to oxytocin was assessed by fluorimetry to detect calcium signals while isolated tracheal rings and precision cut lung slices (PCLS were used to measure contractile responses. Finally, ELISA was used to compare oxytocin levels in the bronchoalveloar lavage (BAL samples from healthy subjects and those with asthma. Results PCR analysis demonstrates that OXTR is expressed in HASMCs under basal conditions and that both interleukin (IL-13 and tumor necrosis factor (TNFα stimulate a time-dependent increase in OXTR expression at 6 and 18 hr. Additionally, oxytocin increases cytosolic calcium levels in fura-2-loaded HASMCs that were enhanced in cells treated for 24 hr with IL-13. Interestingly, TNFα had little effect on oxytocin-induced calcium response despite increasing receptor expression. Using isolated murine tracheal rings and PCLS, oxytocin also promoted force generation and airway narrowing. Further, oxytocin levels are detectable in bronchoalveolar lavage (BAL fluid derived from healthy subjects as well as from those with asthma. Conclusion Taken together, we show that cytokines modulate the expression of functional oxytocin receptors in HASMCs suggesting a

  13. Reporter-free potentiometric sensing of boronic acids and their reactions by using quaternary ammonium salt-functionalized polymeric liquid membranes.

    Science.gov (United States)

    Wang, Xuewei; Yue, Dengfeng; Lv, Enguang; Wu, Lei; Qin, Wei

    2014-02-18

    The tremendous applications of boronic acids (BAs) in chemical sensing, medical chemistry, molecular assembly, and organic synthesis lead to an urgent demand for developing effective sensing methods for BAs. This paper reports a facile and sensitive potentiometric sensor scheme for heterogeneous detection of BAs based on their unexpected potential responses on quaternary ammonium salt-doped polymeric liquid membranes. (11)B NMR data reveal that a quaternary ammonium chloride can trigger the hydrolysis of an electrically neutral BA in an aprotic solvent. Using the quaternary ammonium salt as the receptor, the BA molecules can be extracted from the sample solution into the polymeric membrane phase and undergo the concomitant hydrolysis. Such salt-triggered hydrolysis generates H(+) ions, which can be coejected into the aqueous phase with the counterions (e.g., Cl(-)) owing to their high hydrophilicities. The perturbation on the ionic partition at the sample-membrane interface changes the phase boundary potential and thus enables the potentiometric sensing of BAs. In contrast to other transduction methods for BAs, for which labeled or separate reporters are exclusively required, the present heterogeneous sensing scheme allows the direct detection of BAs without using any reporter molecules. This technique shows superior detection limits for BAs (e.g., 1.0 × 10(-6) M for phenylboronic acid) as compared to previously reported methods based on colorimetry, fluorimetry, and mass spectrometry. The proposed sensing strategy has also been successfully applied to potentiometric indication of the BA reactions with hydrogen peroxide and saccharides, which allows indirect and sensitive detection of these important species.

  14. Comparison of alkali fusion and acid digestion methods for radiochemical separation of Uranium from dietary samples

    International Nuclear Information System (INIS)

    Kamesh Viswanathan, B.; Arunachalam, Kantha D.; Sathesh Kumar, A.; Jayakrishana, K.; Shanmugamsundaram, H.; Rao, D.D.

    2014-01-01

    Several methods exist for separation and measurement of uranium in dietary samples such as neutron activation analysis (NAA), alpha spectrometric determination, inductively coupled plasma mass spectrometry (ICP-MS) and fluorimetry. For qualitative determination of activity, NAA and alpha spectrometry are said to be superior to evaluate the isotopes of uranium ( 238 U, 234 U and 235 U). In case of alpha spectrometry, the samples have to undergo radiochemical analysis for separation from other elements for uranium detection. In our studies, uranium was determined in food matrices by acid digestion (AD) and alkali fusion (AF) methods. The recovery yield of uranium in food matrices was compared in order to get consistent yield. The average activity levels of 238 U and 234 U in food samples were calculated based on recovery yield of 232 U in the samples. The average recovery of 232 U in AD method was 22 ± 8% and in AF method, it was 14.9 ± 1.3%. The spread is more in AD method than the AF method from their mean. The lowest recovery of 232 U was found in AF method. This is due to the interference of other elements in the sample during electroplating. Experimental results showed that the uranium separation by AD method has better recovery than the AF method. The consistency in recovery of 232 U was better for AF method, which was lower than the AD method. However, overall for both the methods, the recovery can be termed as poor and need rigorous follow up studies for consistently higher recoveries (>50%) in these type of biological samples. There are reports indicating satisfactory recoveries of around 80% with 232 U as tracer in the food matrices

  15. Inactivation of Mycobacterium tuberculosis l,d-Transpeptidase LdtMt1 by Carbapenems and Cephalosporins

    Science.gov (United States)

    Dubée, Vincent; Triboulet, Sébastien; Mainardi, Jean-Luc; Ethève-Quelquejeu, Mélanie; Gutmann, Laurent; Marie, Arul; Dubost, Lionel

    2012-01-01

    The structure of Mycobacterium tuberculosis peptidoglycan is atypical since it contains a majority of 3→3 cross-links synthesized by l,d-transpeptidases that replace 4→3 cross-links formed by the d,d-transpeptidase activity of classical penicillin-binding proteins. Carbapenems inactivate these l,d-transpeptidases, and meropenem combined with clavulanic acid is bactericidal against extensively drug-resistant M. tuberculosis. Here, we used mass spectrometry and stopped-flow fluorimetry to investigate the kinetics and mechanisms of inactivation of the prototypic M. tuberculosis l,d-transpeptidase LdtMt1 by carbapenems (meropenem, doripenem, imipenem, and ertapenem) and cephalosporins (cefotaxime, cephalothin, and ceftriaxone). Inactivation proceeded through noncovalent drug binding and acylation of the catalytic Cys of LdtMt1, which was eventually followed by hydrolysis of the resulting acylenzyme. Meropenem rapidly inhibited LdtMt1, with a binding rate constant of 0.08 μM−1 min−1. The enzyme was unable to recover from this initial binding step since the dissociation rate constant of the noncovalent complex was low (carbapenem side chains affected both the binding and acylation steps, ertapenem being the most efficient LdtMt1 inactivator. Cephalosporins also formed covalent adducts with LdtMt1, although the acylation reaction was 7- to 1,000-fold slower and led to elimination of one of the drug side chains. Comparison of kinetic constants for drug binding, acylation, and acylenzyme hydrolysis indicates that carbapenems and cephems can both be tailored to optimize peptidoglycan synthesis inhibition in M. tuberculosis. PMID:22615283

  16. Measurements of natural radioactivity concentration in drinking water samples of Shiraz city and springs of the Fars province, Iran, and dose estimation

    International Nuclear Information System (INIS)

    Mehdizadeh, S.; Faghihi, R.; Sina, S.; Derakhshan, S.

    2013-01-01

    The Fars province is located in the south-west region of Iran where different nuclear sites has been established, such as Bushehr Nuclear Power Plant. In this research, 92 water samples from the water supplies of Shiraz city and springs of the Fars province were investigated with regard to the concentrations of natural radioactive elements, total uranium, 226 Ra, gross alpha and gross beta. 226 Ra concentration was determined by the 222 Rn emanation method. To measure the total uranium concentration, a laser fluorimetry analyzer (UA-3) was used. The mean concentration of 226 Ra in Shiraz's water resources was 23.9 mBq l -1 , while 93% of spring waters have a concentration 2 mBq l -1 . The results of uranium concentration measurements show the mean concentrations of 7.6 and 6 mg l -1 in the water of Shiraz and springs of Fars, respectively. The gross alpha and beta concentrations measured by the evaporation method were lower than the limit of detection of the measuring instruments used in this survey. The mean annual effective doses of infants, children and adults from 238 U and 226 Ra content of Shiraz's water and spring waters were estimated. According to the results of this study, the activity concentration in water samples were below the maximum permissible concentrations determined by the World Health Organization and the US Environmental Protection Agency. Finally, the correlation between 226 Ra and total U activity concentrations and geochemical properties of water samples, i.e. pH, total dissolve solids and SO 4 2- , were estimated. (authors)

  17. Maternal intake of trans-unsaturated or interesterified fatty acids during pregnancy and lactation modifies mitochondrial bioenergetics in the liver of adult offspring in mice.

    Science.gov (United States)

    de Velasco, Patricia C; Chicaybam, Gustavo; Ramos-Filho, Dionizio M; Dos Santos, Raísa M A R; Mairink, Caroline; Sardinha, Fátima L C; El-Bacha, Tatiana; Galina, Antonio; Tavares-do-Carmo, Maria das Graças

    2017-07-01

    The quality of dietary lipids in the maternal diet can programme the offspring to diseases in later life. We investigated whether the maternal intake of palm oil or interesterified fat, substitutes for trans-unsaturated fatty acids (FA), induces metabolic changes in the adult offspring. During pregnancy and lactation, C57BL/6 female mice received normolipidic diets containing partially hydrogenated vegetable fat rich in trans-unsaturated fatty acids (TG), palm oil (PG), interesterified fat (IG) or soyabean oil (CG). After weaning, male offspring from all groups received the control diet until day 110. Plasma glucose and TAG and liver FA profiles were ascertained. Liver mitochondrial function was accessed with high-resolution respirometry by measuring VO2, fluorimetry for detection of hydrogen peroxide (H2O2) production and mitochondrial Ca2+ uptake. The results showed that the IG offspring presented a 20 % increase in plasma glucose and both the IG and TG offspring presented a 2- and 1·9-fold increase in TAG, respectively, when compared with CG offspring. Liver MUFA and PUFA contents decreased in the TG and IG offspring when compared with CG offspring. Liver MUFA content also decreased in the PG offspring. These modifications in FA composition possibly affected liver mitochondrial function, as respiration was impaired in the TG offspring and H2O2 production was higher in the IG offspring. In addition, mitochondrial Ca2+ retention capacity was reduced by approximately 40 and 55 % in the TG and IG offspring, respectively. In conclusion, maternal consumption of trans-unsaturated and interesterified fat affected offspring health by compromising mitochondrial bioenergetics and lipid metabolism in the liver.

  18. Genipin-induced inhibition of uncoupling protein-2 sensitizes drug-resistant cancer cells to cytotoxic agents.

    Directory of Open Access Journals (Sweden)

    Ryan J Mailloux

    2010-10-01

    Full Text Available Uncoupling protein-2 (UCP2 is known to suppress mitochondrial reactive oxygen species (ROS production and is employed by drug-resistant cancer cells to mitigate oxidative stress. Using the drug-sensitive HL-60 cells and the drug-resistant MX2 subline as model systems, we show that genipin, a UCP2 inhibitor, sensitizes drug-resistant cells to cytotoxic agents. Increased MX2 cell death was observed upon co-treatment with genipin and different doses of menadione, doxorubicin, and epirubicin. DCFH-DA fluorimetry revealed that the increase in MX2 cell death was accompanied by enhanced cellular ROS levels. The drug-induced increase in ROS was linked to genipin-mediated inhibition of mitochondrial proton leak. State 4 and resting cellular respiratory rates were higher in the MX2 cells in comparison to the HL-60 cells, and the increased respiration was readily suppressed by genipin in the MX2 cells. UCP2 accounted for a remarkable 37% of the resting cellular oxygen consumption indicating that the MX2 cells are functionally reliant on this protein. Higher amounts of UCP2 protein were detected in the MX2 versus the HL-60 mitochondria. The observed effects of genipin were absent in the HL-60 cells pointing to the selectivity of this natural product for drug-resistant cells. The specificity of genipin for UCP2 was confirmed using CHO cells stably expressing UCP2 in which genipin induced an ∼22% decrease in state 4 respiration. These effects were absent in empty vector CHO cells expressing no UCP2. Thus, the chemical inhibition of UCP2 with genipin sensitizes multidrug-resistant cancer cells to cytotoxic agents.

  19. Polarographic determination of Sn (II) and total Sn in PYRO and MDP radiopharmaceutical kits

    International Nuclear Information System (INIS)

    Sebastian, Maria V.A.; Lugon, Marcelo Di M.V.; Silva, Jose L. da; Fukumori, Neuza T.O.; Pereira, Nilda P.S. de; Silva, Constancia P.G. da; Matsuda, Margareth M.N.

    2007-01-01

    A sensitive, alternative method to atom absorption spectrometry, fluorimetry or potentiometry for the evaluation of tin(II) ions (0.1- 10 mg) and total tin in radiopharmaceutical kits was investigated. Differential pulse polarography was chosen. The supporting electrolyte was H 2 SO 4 3 mol L -1 and HCl 3 mol L -1 solution. The potential was swept from -250 to -800 mV vs Ag/AgCl/saturated KCl, using a dropping mercury electrode with 1 s drop time, 50 mV s -1 scan rate, -50 mV pulse amplitude, 40 ms pulse time and 10 mV step amplitude. Pure nitrogen was used to deaerate the polarographic cell solution for 5 min, before and after each sample introduction. Oxidation of Sn(II) was made in the same sample vial by adding H 2 O 2 (hydrogen peroxide) 10 mol L -1 , at 37 deg C, in order to quantify the total Sn. The calibration curve for Sn(II) and Sn(IV) was obtained in the concentration range of 0-10 ppm from a 1000 ppm standard solution. The detection limit of Sn(II) is 0.5 ppm and for Sn(IV) is 0.6 ppm. Differential pulse polarography was performed in the pyrophosphate (PYRO) and methylenediphosphonic acid (MDP) radiopharmaceutical kits, containing 2 mg and 1 mg of SnCl 2 .2H 2 O per vial, respectively. The described method for determination of stannous ion (Sn(II)), is selective, reproducible and adequate to be used in the quality control of lyophilized reagents and it shall be performed for other cold kits produced at IPEN. (author)

  20. A survey of natural uranium concentrations in drinking water supplies in Iran

    International Nuclear Information System (INIS)

    Alirezazadeh, N.; Garshasbi, N.

    2003-01-01

    Background: Measurement of background concentration of uranium in drinking water is very important for many reasons, specially, for human health. The uranium concentration in drinking water in many countries is a matter of concern for clinical and radioactive poisoning. Materials and methods: The uranium concentration in drinking water is determined using laser fluorimetric uranium analyzer. For this purpose after sampling, sample handling and sample preserving, sample preparation and treatment for reduction of organic matter, the concentration of uranium is measured. Results: To determine the uranium concentrations in drinking water in Iran, nearly 200 water samples were collected from all sources supplying drinking water in 21 provincial centers in the country. The wells were found to be the main source for drinking water. Uranium in the samples was measured by a laser fluorimetry technique. According to results, the concentration values found in the wells ranged from 1.0 to 10.90 μgL -1 , while nearly 95 percent of the cities had uranium concentrations in the wells at less than 4.70 μgL -1 . Surface waters showed uranium concentrations in the range of 0.75 to 2.58 μgL -1 . The daily intake of uranium from drinking water was estimated to range from 2.04 to 21.80 μgd -1 , with the mean value of 5.44 μgd -1 . Conclusion: Highest uranium mean concentration of 10.9 μgL -1 was found in Ardabil area where more studies should be done in that province in the future

  1. The Tp0684 (MglB-2 Lipoprotein of Treponema pallidum: A Glucose-Binding Protein with Divergent Topology.

    Directory of Open Access Journals (Sweden)

    Chad A Brautigam

    Full Text Available Treponema pallidum, the bacterium that causes syphilis, is an obligate human parasite. As such, it must acquire energy, in the form of carbon sources, from the host. There is ample evidence that the principal source of energy for this spirochete is D-glucose acquired from its environment, likely via an ABC transporter. Further, there is genetic evidence of a D-glucose chemotaxis system in T. pallidum. Both of these processes may be dependent on a single lipidated chemoreceptor: Tp0684, also called TpMglB-2 for its sequence homology to MglB of Escherichia coli. To broaden our understanding of this potentially vital protein, we determined a 2.05-Å X-ray crystal structure of a soluble form of the recombinant protein. Like its namesake, TpMglB-2 adopts a bilobed fold that is similar to that of the ligand-binding proteins (LBPs of other ABC transporters. However, the protein has an unusual, circularly permuted topology. This feature prompted a series of biophysical studies that examined whether the protein's topological distinctiveness affected its putative chemoreceptor functions. Differential scanning fluorimetry and isothermal titration calorimetry were used to confirm that the protein bound D-glucose in a cleft between its two lobes. Additionally, analytical ultracentrifugation was employed to reveal that D-glucose binding is accompanied by a significant conformational change. TpMglB-2 thus appears to be fully functional in vitro, and given the probable central importance of the protein to T. pallidum's physiology, our results have implications for the viability and pathogenicity of this obligate human pathogen.

  2. The Tp0684 (MglB-2) Lipoprotein of Treponema pallidum: A Glucose-Binding Protein with Divergent Topology.

    Science.gov (United States)

    Brautigam, Chad A; Deka, Ranjit K; Liu, Wei Z; Norgard, Michael V

    2016-01-01

    Treponema pallidum, the bacterium that causes syphilis, is an obligate human parasite. As such, it must acquire energy, in the form of carbon sources, from the host. There is ample evidence that the principal source of energy for this spirochete is D-glucose acquired from its environment, likely via an ABC transporter. Further, there is genetic evidence of a D-glucose chemotaxis system in T. pallidum. Both of these processes may be dependent on a single lipidated chemoreceptor: Tp0684, also called TpMglB-2 for its sequence homology to MglB of Escherichia coli. To broaden our understanding of this potentially vital protein, we determined a 2.05-Å X-ray crystal structure of a soluble form of the recombinant protein. Like its namesake, TpMglB-2 adopts a bilobed fold that is similar to that of the ligand-binding proteins (LBPs) of other ABC transporters. However, the protein has an unusual, circularly permuted topology. This feature prompted a series of biophysical studies that examined whether the protein's topological distinctiveness affected its putative chemoreceptor functions. Differential scanning fluorimetry and isothermal titration calorimetry were used to confirm that the protein bound D-glucose in a cleft between its two lobes. Additionally, analytical ultracentrifugation was employed to reveal that D-glucose binding is accompanied by a significant conformational change. TpMglB-2 thus appears to be fully functional in vitro, and given the probable central importance of the protein to T. pallidum's physiology, our results have implications for the viability and pathogenicity of this obligate human pathogen.

  3. Evaluation of the radiological impact in the aquatic system in the surrounding of IPEN-CNEN/SP

    International Nuclear Information System (INIS)

    Jacomino, V.M.F.; Gordon, A.M.P.L.; Venturini, L.

    1989-12-01

    In order to control the discharges of radioactive material in the aquatic system (Jaguare stream and Pinheiros river) in the vicinity of IPEN-CNEN/SP an effluent monitoring program was established on a routine basis. This control is carried out by measuring the activity of the radionuclides present in the liquid effluents (source term) using gamma spectrometry and/or spectrophotometry. The results obtained are then compared with the discharge limits adopted by CNEN when a decision is made upon the discharge of the effluent under consideration. In 1988 the total activity discharged by the nuclear installations available was 1997, 9x10 6 Bq in a total volume of 2421,4 m 3 . The next step was to evaluate the effective equivalent dose in the general public by using the results of the source term and the information available concerning the environment. It was assumed that the only critical pathway is the external gamma irradiation of the people that work in the nearby of the discharge points in Pinheiros river. The effective equivalent dose obtained was 39,4η Sv and the most relevant radionuclides that should be considered 60 Co, 137 Cs, 131 I and 226 Ra. This result is less than 1/10 of the maximum admissible dose limit adopted by the Radiological Protection Standards which is 10 -3 Sv/year. In order to measure the level of radioactivity in the environment, samples of ground water, water and sediments of Pinheiros river were collected and analyzed by using gamma spectrometry and fluorimetry. The radionuclides found were 226 Ra, 22228 Ra, U Nat, 40 K and 7 Be with activities corresponding to the background levels. The analysis of these results and the evaluation of the equivalent dose show that the radiological impact in the aquatic systems considered is insignificant. (author) [pt

  4. Radioactivity content of Shiraz water supplies and its related effective dose

    International Nuclear Information System (INIS)

    Mehdizadeh, S.; Derakhshan, Sh.

    2004-01-01

    We are exposed naturally to ionizing radiation from cosmic rays and natural radionuclides in the air, ground, food and drinking water. When invested, naturally and radionuclides are distributed among body organs. The average concentrations of naturally occurring radionuclides in drinking water are used to estimated the annual effective dose equivalents. In this work, during October of 2002 to October of 2003, 37 samples from 29 different wells and Doroodzan dam were gathered in winter and summer and the gross Alfa and Beta, total uranium and Ra-226 activity in each sample were determined. The activity of Ra-226 was measured by Radon Emanation method and total uranium was determined using laser Fluorimetry. The mean concentration of Ra-226 was higher in summer but the uranium had higher concentration in winter. The mean concentration of Ra-226 and Uranium were 15.72±1.12 mBq.1 -1 and 9.44±1.14μg.1 -1 in winter ±μ and 29.46 2.18 mBq.1 -1 and 7.37 1.1 g.1 -1 in summer respectively. The average annual effective doses (Ra-226) of different age groups (Adults, children and infants) were equal to 2.37± 0.22, 5.42 ±0.43, 2.66±0.24, mSv.y 1 -1 and 0.136±0.02, 0.18±0.027 and 0.171±0.026 mSv.y -1 due to existence of uranium in drinking water. The results of this survey showed that the radioactive concentration of the radionuclides and their related effective doses are in the normal range and permissible levels. These results are comparable with those found by Atomic Energy Organization of Iran but different with the results of the limited project done during 1999 at Shiraz

  5. Radio-ecological study of the Lodeve mining complex (France) 1981-1985

    International Nuclear Information System (INIS)

    Descamps, B.

    1987-11-01

    The radio-ecological study of the Lodeve mining complex (mine + uranium process plant) was carried out between 1981 and 1985. Four aquatic compartments -water, sediments, vegetals, fish- were studied in eight stations selected on the basis of the existence of two liquid wastes. The measurements essentially concerned radium 226 and uranium 238 but also lead 210 (1985), thorium 232, potassium 40 and cesium 137. Spectrometry γ Ge-Li, emanometry and fluorimetry were used. Only a small brook -Rivernoux- an affluent of the Lergue river, shows radium and uranium activity levels higher than those measured upstream of the site; and this for the four compartments. In water, radium is associated with the solved cationic fraction (66%) and with materials in suspension (21%), whereas uranium is essentially associated with the solved anionic fraction (84%); the radium migration potentialities are therefore lesser than that of uranium. Radium distribution in fish is as follows: flesh (5 to 15%), skeleton (12%), viscera (30%) and skin + fins (30%). Radium concentration (or exchange) factors are always higher than those corresponding to uranium. In general, they are more important in non-influenced zones than in influenced zones (Rivernoux); this implies that radium and uranium evacuated by the mining complex are, at least partly, in non bio-available forms. For water/fish exchange a concentration factor of 100 will be considered for radium. On the basis of this value a weekly consumption of 200 g of fried gudgeon gives an annual committed dose equivalent corresponding to 0.86% of the dose authorized for the public (5.10 -3 Sv.an -1 ) [fr

  6. The fitness for purpose of analytical methods applied to fluorimetric uranium determination in water matrix

    International Nuclear Information System (INIS)

    Grinman, Ana; Giustina, Daniel; Mondini, Julia; Diodat, Jorge

    2008-01-01

    Full text: This paper describes the steps which should be followed by a laboratory in order to validate the fluorimetric method for natural uranium in water matrix. The validation of an analytical method is a necessary requirement prior accreditation under Standard norm ISO/IEC 17025, of a non normalized method. Different analytical techniques differ in a sort of variables to be validated. Depending on the chemical process, measurement technique, matrix type, data fitting and measurement efficiency, a laboratory must set up experiments to verify reliability of data, through the application of several statistical tests and by participating in Quality Programs (QP) organized by reference laboratories such as the National Institute of Standards and Technology (NIST), National Physics Laboratory (NPL), or Environmental Measurements Laboratory (EML). However, the participation in QP not only involves international reference laboratories, but also, the national ones which are able to prove proficiency to the Argentinean Accreditation Board. The parameters that the ARN laboratory had to validate in the fluorimetric method to fit in accordance with Eurachem guide and IUPAC definitions, are: Detection Limit, Quantification Limit, Precision, Intra laboratory Precision, Reproducibility Limit, Repeatability Limit, Linear Range and Robustness. Assays to fit the above parameters were designed on the bases of statistics requirements, and a detailed data treatment is presented together with the respective tests in order to show the parameters validated. As a final conclusion, the uranium determination by fluorimetry is a reliable method for direct measurement to meet radioprotection requirements in water matrix, within its linear range which is fixed every time a calibration is carried out at the beginning of the analysis. The detection limit ( depending on blank standard deviation and slope) varies between 3 ug U and 5 ug U which yields minimum detectable concentrations (MDC) of

  7. In vitro bioassay investigations of the endocrine disrupting potential of steviol glycosides and their metabolite steviol, components of the natural sweetener Stevia.

    Science.gov (United States)

    Shannon, Maeve; Rehfeld, Anders; Frizzell, Caroline; Livingstone, Christina; McGonagle, Caoimhe; Skakkebaek, Niels E; Wielogórska, Ewa; Connolly, Lisa

    2016-05-15

    The food industry is moving towards the use of natural sweeteners such as those produced by Stevia rebaudiana due to the number of health and safety concerns surrounding artificial sweeteners. Despite the fact that these sweeteners are natural; they cannot be assumed safe. Steviol glycosides have a steroidal structure and therefore may have the potential to act as an endocrine disruptor in the body. Reporter gene assays (RGAs), H295R steroidogenesis assay and Ca(2+) fluorimetry based assays using human sperm cells have been used to assess the endocrine disrupting potential of two steviol glycosides: stevioside and rebaudioside A, and their metabolite steviol. A decrease in transcriptional activity of the progestagen receptor was seen following treatment with 25,000 ng/ml steviol in the presence of progesterone (157 ng/ml) resulting in a 31% decrease in progestagen response (p=<0.01). At the level of steroidogenesis, the metabolite steviol (500-25,000 ng/ml) increased progesterone production significantly by 2.3 fold when exposed to 10,000 ng/ml (p=<0.05) and 5 fold when exposed to 25,000 ng/ml (p=<0.001). Additionally, steviol was found to induce an agonistic response on CatSper, a progesterone receptor of sperm, causing a rapid influx of Ca(2+). The response was fully inhibited using a specific CatSper inhibitor. These findings highlight the potential for steviol to act as a potential endocrine disruptor. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

  8. Passive membrane penetration by ZnO nanoparticles is driven by the interplay of electrostatic and phase boundary conditions.

    Science.gov (United States)

    Tiwari, Anuj; Prince, Ashutosh; Arakha, Manoranjan; Jha, Suman; Saleem, Mohammed

    2018-02-15

    The internalization of nanoparticles through the biological membrane is of immense importance for biomedical applications. A fundamental understanding of the lipid specificity and the role of the membrane biochemical and physical forces at play in modulating penetration are lacking. The current understanding of nanoparticle-membrane interaction is drawn mostly from computational studies and lacks sufficient experimental evidence. Herein, using confocal fluorescence imaging and potentiometric dye-based fluorimetry, we first investigated the interaction of ZnONP in both multi-component and individual lipid membranes using cell-like giant unilamellar vesicles to dissect the lipid specificity; also, we investigated the changes in membrane order, anisotropy and hydrophobicity. ZnONP was found to interact with phosphatidylinositol and phosphatidylcholine head-group-containing lipids specifically. We further investigated the interaction of ZnONP with three physiologically relevant membrane conditions varying in composition and dipole potential. We found that ZnONP interaction leads to a photoinduced enhancement of the partial-to-complete phase separation depending upon the membrane composition and cholesterol content. Interestingly, while the lipid order of a partially-phase-separated membrane remained unchanged upon ZnONP crowding, a fully-phase-separated membrane showed an increase in the lipid order. Strikingly, ZnONP crowding induced a contrasting effect on the fluorescence anisotropy of the membrane upon binding to the two membrane conditions, in line with the measured diffusion coefficient. ZnONP seems to preferentially penetrate through the liquid disordered areas of the membrane and the boundaries of the phase-separated regions driven by the interplay between the electrostatics and phase boundary conditions, which are collectively dictated by the composition and ZnONP-induced lipid reorganization. The results may lead to a greater understanding of the interplay of

  9. Assessment and evaluation of speciation tools for the study of uranium (6) circulating biological species

    International Nuclear Information System (INIS)

    Scapolan, St.

    1998-01-01

    The aim of this work is the development of tools allowing the study of uranium VI speciation in inorganic and organic environments, and in particular in the blood environment. The characterization of the different complexed forms of the uranyl ion has been improved by combining two techniques: the time resolution laser spectro-fluorimetry (TRLS) and the capillary electrophoresis (CE). CE is a developing separative analytical technique with a strong resolution. Therefore, the following studies have been carried out: the analysis of the speciation of hydroxo complexes of the uranyl ion by indirect UV detection, with the qualitative identification of the (UO 2 ) 2 (OH) 2 2+ and (UO 2 ) 3 (OH) 5 + complexes; the application of the iso-electrical focussing mode in order to show the uranium-transferrin complexing; the evaluation of the electrophoretic mobility of UO 2 2+ and the interactions with the phosphate, hydroxo-isobutyric acid (HIBA) and citrate ligands; and the study of the U(VI)-blood serum system with the separation of the different seric proteins and the influence of U(VI) on electrophoretic profiles. two points are important to consider in the development of the system: the coupling with an on-line detector (mass spectrometer, counter, laser) and the surface grafting of the capillaries used. The TRLS is a system used in geology, in the Purex process chemistry, in medical and environmental control, and in nuclear wastes management. After having analyzed the influence of different ligands (citrate, phosphate, carbonate, transferrin) in the fluorescence spectra of uranium (VI), the complexing conditional constant (K) of the U(VI)-transferrin system has been evaluated for the first time using a mathematical model and the titration. A study performed on blood plasma has permitted to show a U(VI)-phosphates complexing and finally, three phosphate complexes UO 2 H 2 PO 4 + , UO 2 HPO 4 and UO 2 PO 4 + have been characterized both spectrally and temporarily. (J.S.)

  10. Preparation of MS2 phage-like particles and their use as potential process control viruses for detection and quantification of enteric RNA viruses in different matrices

    Directory of Open Access Journals (Sweden)

    Pavel Mikel

    2016-12-01

    Full Text Available The detection and quantification of enteric RNA viruses is based on isolation of viral RNA from the sample followed by quantitative reverse transcription polymerase chain reaction (RT-qPCR. To control the whole process of analysis and in order to guarantee the validity and reliability of results, process control viruses (PCV are used. The present article describes the process of preparation and use of such PCV– MS2 phage-like particles (MS2 PLP – in RT-qPCR detection and quantification of enteric RNA viruses. The MS2 PLP were derived from bacteriophage MS2 carrying a unique and specific de novo-constructed RNA target sequence originating from the DNA of two extinct species. The amount of prepared MS2 particles was quantified using four independent methods - UV spectrophotometry, fluorimetry, transmission electron microscopy (TEM and a specifically developed duplex RT-qPCR. To evaluate the usefulness of MS2 PLP in routine diagnostics different matrices known to harbor enteric RNA viruses (swab samples, liver tissue, serum, feces, and vegetables were artificially contaminated with specific amounts of MS2 PLP. The extraction efficiencies were calculated for each individual matrix. The prepared particles fulfill all requirements for PCV – they are very stable, non-infectious, and are genetically distinct from the target RNA viruses. Due to these properties they represent a good morphological and physiochemical model. The use of MS2 PLP as a PCV in detection and quantification of enteric RNA viruses was evaluated in different types of matrices.

  11. Changes in the stability and biomechanics of P22 bacteriophage capsid during maturation.

    Science.gov (United States)

    Kant, Ravi; Llauró, Aida; Rayaprolu, Vamseedhar; Qazi, Shefah; de Pablo, Pedro J; Douglas, Trevor; Bothner, Brian

    2018-03-15

    The capsid of P22 bacteriophage undergoes a series of structural transitions during maturation that guide it from spherical to icosahedral morphology. The transitions include the release of scaffold proteins and capsid expansion. Although P22 maturation has been investigated for decades, a unified model that incorporates thermodynamic and biophysical analyses is not available. A general and specific model of icosahedral capsid maturation is of significant interest to theoreticians searching for fundamental principles as well as virologists and material scientists seeking to alter maturation to their advantage. To address this challenge, we have combined the results from orthogonal biophysical techniques including differential scanning fluorimetry, atomic force microscopy, circular dichroism, and hydrogen-deuterium exchange mass spectrometry. By integrating these results from single particle and population measurements, an energy landscape of P22 maturation from procapsid through expanded shell to wiffle ball emerged, highlighting the role of metastable structures and the thermodynamics guiding maturation. The propagation of weak quaternary interactions across symmetric elements of the capsid is a key component for stability in P22. A surprising finding is that the progression to wiffle ball, which lacks pentamers, shows that chemical and thermal stability can be uncoupled from mechanical rigidity, elegantly demonstrating the complexity inherent in capsid protein interactions and the emergent properties that can arise from icosahedral symmetry. On a broader scale, this work demonstrates the power of applying orthogonal biophysical techniques to elucidate assembly mechanisms for supramolecular complexes and provides a framework within which other viral systems can be compared. Copyright © 2018 Elsevier B.V. All rights reserved.

  12. New insight into silica deposition in horsetail (Equisetum arvense

    Directory of Open Access Journals (Sweden)

    Exley Christopher

    2011-07-01

    Full Text Available Abstract Background The horsetails (Equisetum sp are known biosilicifiers though the mechanism underlying silica deposition in these plants remains largely unknown. Tissue extracts from horsetails grown hydroponically and also collected from the wild were acid-digested in a microwave oven and their silica 'skeletons' visualised using the fluor, PDMPO, and fluorescence microscopy. Results Silica deposits were observed in all plant regions from the rhizome through to the stem, leaf and spores. Numerous structures were silicified including cell walls, cell plates, plasmodesmata, and guard cells and stomata at varying stages of differentiation. All of the major sites of silica deposition in horsetail mimicked sites and structures where the hemicellulose, callose is known to be found and these serendipitous observations of the coincidence of silica and callose raised the possibility that callose might be templating silica deposition in horsetail. Hydroponic culture of horsetail in the absence of silicic acid resulted in normal healthy plants which, following acid digestion, showed no deposition of silica anywhere in their tissues. To test the hypothesis that callose might be templating silica deposition in horsetail commercially available callose was mixed with undersaturated and saturated solutions of silicic acid and the formation of silica was demonstrated by fluorimetry and fluorescence microscopy. Conclusions The initiation of silica formation by callose is the first example whereby any biomolecule has been shown to induce, as compared to catalyse, the formation of silica in an undersaturated solution of silicic acid. This novel discovery allowed us to speculate that callose and its associated biochemical machinery could be a missing link in our understanding of biosilicification.

  13. Complexation modeling of uranium and other actinides by organic compounds of natural or synthetic origin

    International Nuclear Information System (INIS)

    Bouby, M.

    1998-01-01

    The behaviour of nuclear wastes raises many questions, the answers of which require a precise knowledge of the physical, chemical and biological processes affecting the properties of the radio-elements present in the wastes. Three ways of research are approached. The results obtained give some elements of answer to these questions. The experimental methods that have been used are the neutron activation analysis, the UV-visible spectrophotometry, and the time-resolved laser spectro-fluorimetry. The analysis of the results has permitted to model part of the phenomena evidenced by using suitable ionic force correction models (Davies or MSA type) when chemical equilibria have been considered. The main results show: the bio-sorption capacities of Mycobacterium phlei microorganism with respect to UO 2 2+ and NpO 2+ cations such as: Q eq (UO 2 2+ ) = 60 and Q eq (NpO 2+ ) = 444 moles of cation per g of dry biomass; the retention capacities, in various leaching conditions, of this bacteria of the preliminarily adsorbed ions; the complexation properties of two siderophores with respect to UO 2 2+ , U 4+ and Th 4+ cations. One siderophore, Pyoverdine A, shows a selectiveness which is explained by the value of the thermodynamic equilibrium constant determined for each cation using the same model: K(UO 2 2+ ) 4+ ) 4+ ). The behaviour in highly acid environment (HCl and HClO 4 up to 12 M) of acylisoxazolone HPBI (1-phenyl-4-benzoyl-5-isoxazolone) and the value of its acidity thermodynamical constant (0.13 th 4 and CF 3 SO 3 H up to 12 M). It seems that a complexation between uranyl and the counter-ions present in the solution occurs. (J.S.)

  14. Maltase protein of Ogataea (Hansenula) polymorpha is a counterpart to the resurrected ancestor protein ancMALS of yeast maltases and isomaltases

    Science.gov (United States)

    Viigand, Katrin; Visnapuu, Triinu; Mardo, Karin; Aasamets, Anneli

    2016-01-01

    Abstract Saccharomyces cerevisiae maltases use maltose, maltulose, turanose and maltotriose as substrates, isomaltases use isomaltose, α‐methylglucoside and palatinose and both use sucrose. These enzymes are hypothesized to have evolved from a promiscuous α‐glucosidase ancMALS through duplication and mutation of the genes. We studied substrate specificity of the maltase protein MAL1 from an earlier diverged yeast, Ogataea polymorpha (Op), in the light of this hypothesis. MAL1 has extended substrate specificity and its properties are strikingly similar to those of resurrected ancMALS. Moreover, amino acids considered to determine selective substrate binding are highly conserved between Op MAL1 and ancMALS. Op MAL1 represents an α‐glucosidase in which both maltase and isomaltase activities are well optimized in a single enzyme. Substitution of Thr200 (corresponds to Val216 in S. cerevisiae isomaltase IMA1) with Val in MAL1 drastically reduced the hydrolysis of maltose‐like substrates (α‐1,4‐glucosides), confirming the requirement of Thr at the respective position for this function. Differential scanning fluorimetry (DSF) of the catalytically inactive mutant Asp199Ala of MAL1 in the presence of its substrates and selected monosaccharides suggested that the substrate‐binding pocket of MAL1 has three subsites (–1, +1 and +2) and that binding is strongest at the –1 subsite. The DSF assay results were in good accordance with affinity (K m) and inhibition (K i) data of the enzyme for tested substrates, indicating the power of the method to predict substrate binding. Deletion of either the maltase (MAL1) or α‐glucoside permease (MAL2) gene in Op abolished the growth of yeast on MAL1 substrates, confirming the requirement of both proteins for usage of these sugars. © 2016 The Authors. Yeast published by John Wiley & Sons, Ltd. PMID:26919272

  15. Molecular characterization of dissolved organic matter (DOM): a critical review.

    Science.gov (United States)

    Nebbioso, Antonio; Piccolo, Alessandro

    2013-01-01

    Advances in water chemistry in the last decade have improved our knowledge about the genesis, composition, and structure of dissolved organic matter, and its effect on the environment. Improvements in analytical technology, for example Fourier-transform ion cyclotron (FT-ICR) mass spectrometry (MS), homo and hetero-correlated multidimensional nuclear magnetic resonance (NMR) spectroscopy, and excitation emission matrix fluorimetry (EEMF) with parallel factor (PARAFAC) analysis for UV-fluorescence spectroscopy have resulted in these advances. Improved purification methods, for example ultrafiltration and reverse osmosis, have enabled facile desalting and concentration of freshly collected DOM samples, thereby complementing the analytical process. Although its molecular weight (MW) remains undefined, DOM is described as a complex mixture of low-MW substances and larger-MW biomolecules, for example proteins, polysaccharides, and exocellular macromolecules. There is a general consensus that marine DOM originates from terrestrial and marine sources. A combination of diagenetic and microbial processes contributes to its origin, resulting in refractory organic matter which acts as carbon sink in the ocean. Ocean DOM is derived partially from humified products of plants decay dissolved in fresh water and transported to the ocean, and partially from proteinaceous and polysaccharide material from phytoplankton metabolism, which undergoes in-situ microbial processes, becoming refractory. Some of the DOM interacts with radiation and is, therefore, defined as chromophoric DOM (CDOM). CDOM is classified as terrestrial, marine, anthropogenic, or mixed, depending on its origin. Terrestrial CDOM reaches the oceans via estuaries, whereas autochthonous CDOM is formed in sea water by microbial activity; anthropogenic CDOM is a result of human activity. CDOM also affects the quality of water, by shielding it from solar radiation, and constitutes a carbon sink pool. Evidence in support

  16. Supramolecular solvent-based extraction of benzimidazolic fungicides from natural waters prior to their liquid chromatographic/fluorimetric determination.

    Science.gov (United States)

    Moral, Antonia; Sicilia, María Dolores; Rubio, Soledad

    2009-05-01

    A supramolecular solvent made up of vesicles of decanoic acid in the nano- and microscale regimes dispersed in a continuous aqueous phase is proposed for the extraction/preconcentration of benzimidazolic fungicides (BFs) from river and underground water samples prior to their determination by liquid chromatography (LC)/fluorimetry. The solvent is produced from the coacervation of decanoic acid aqueous vesicles by the action of tetrabutylammonium (Bu(4)N(+)). Carbendazim (CB), thiabendazole (TB) and fuberidazole (FB) are extracted on the basis of hydrophobic and pi-cation interactions and the formation of hydrogen bonds. The extraction provides high preconcentration factors (160 for CB and 190 for TB and FB), requires a short time (the procedure takes less than 20 min and several samples can be simultaneously processed) and a low sample volume (20 mL), and avoids the use of toxic organic solvents. Because of the absence of matrix interferences and the low viscosity of the extracts, these can be directly injected into the chromatographic system without the need of cleaning-up or diluting them. Recoveries are not influenced by the presence of salt concentrations up to 1 M. The proposed method provides detection limits for the determination of CB, TB and FB in natural waters of 32, 4 and 0.1 ng L(-1), respectively, and a precision, expressed as relative standard deviation (n=11) of 5.5% for CB (100 ng L(-1)), 4.0% for TB (80 ng L(-1)) and 2.5% for FB (30 ng L(-1)). Recoveries obtained by applying this approach to the analysis of river and underground water samples fortified at the ng L(-1) level are in the intervals 75-83, 95-102 and 97-101% for CB, TB and FB, respectively.

  17. Studies on effective decomposition of monazite minerals by variety of phosphate fluxes for simple and direct determination of uranium by LED fluorimeter

    International Nuclear Information System (INIS)

    Gopal, Leela; Hanuman, V.V.; Chakrapani, G.

    2013-01-01

    A simple, rapid, effective sample decomposition method is developed for the determination of uranium (U) in monazite minerals by fluorimetric (Light Emitting Diodes (LED) based) technique. The salts of sodium dihydrogen phosphate (NaH 2 PO 4 ), disodium hydrogen phosphate (Na 2 HPO 4 ) and tetrasodium pyrophosphate (Na 4 P 2 O 7 ) were used to conduct studies on effective decomposition and dissolution of monazite minerals. The flux short listed for sample decomposition has several advantages. The fusion is very simple (involve minimal skills), time saving and eco-friendly (no acids were used for sample dissolution), where as in the reported conventional sample decomposition methods involving fusion with sodium peroxide, mixture of KHF 2 and NaF, mineral acids are being used for sample decomposition to get clear solution. Further this solution cannot be used directly for uranium determination by LED fluorimetry; hence separation is required, resulting in low sample throughput. In the present method no such separation is required as the flux itself acts as a fluorescence enhancing reagent and buffer (maintaining the optimum pH of 7.1 ± 0.1). The fused melt of the flux mixture when dissolved in water, gives clear and stable solution. The accuracy and precision of the method was evaluated by analyzing Certified Reference Material, IGS-36 (Institute of Geological Sciences, UK) and monazite samples received from BSOI, Trivandrum. The accuracy of the data was further evaluated by comparing with conventional standard decomposition methods. The results are well within experimental error. RSD of the method is ±2% at 0.30% U 3 O 8 in monazite minerals. (author)

  18. SAXS and stability studies of iron-induced oligomers of bacterial frataxin CyaY.

    Directory of Open Access Journals (Sweden)

    Mostafa Fekry

    Full Text Available Frataxin is a highly conserved protein found in both prokaryotes and eukaryotes. It is involved in several central functions in cells, which include iron delivery to biochemical processes, such as heme synthesis, assembly of iron-sulfur clusters (ISC, storage of surplus iron in conditions of iron overload, and repair of ISC in aconitase. Frataxin from different organisms has been shown to undergo iron-dependent oligomerization. At least two different classes of oligomers, with different modes of oligomer packing and stabilization, have been identified. Here, we continue our efforts to explore the factors that control the oligomerization of frataxin from different organisms, and focus on E. coli frataxin CyaY. Using small-angle X-ray scattering (SAXS, we show that higher iron-to-protein ratios lead to larger oligomeric species, and that oligomerization proceeds in a linear fashion as a results of iron oxidation. Native mass spectrometry and online size-exclusion chromatography combined with SAXS show that a dimer is the most common form of CyaY in the presence of iron at atmospheric conditions. Modeling of the dimer using the SAXS data confirms the earlier proposed head-to-tail packing arrangement of monomers. This packing mode brings several conserved acidic residues into close proximity to each other, creating an environment for metal ion binding and possibly even mineralization. Together with negative-stain electron microscopy, the experiments also show that trimers, tetramers, pentamers, and presumably higher-order oligomers may exist in solution. Nano-differential scanning fluorimetry shows that the oligomers have limited stability and may easily dissociate at elevated temperatures. The factors affecting the possible oligomerization mode are discussed.

  19. What role does measuring medication compliance play in evaluating the efficacy of naltrexone?

    Science.gov (United States)

    Baros, Alicia M; Latham, Patricia K; Moak, Darlene H; Voronin, Konstantin; Anton, Raymond F

    2007-04-01

    Compliance with medication in pharmacotherapy trials of alcoholism has been shown to be equal to, or more, important than in other areas of medicine. Research has suggested that naltrexone's effectiveness can be greatly influenced by the compliance of participants in clinical trials. Presently, we compare 2 compliance measurement methods [urine riboflavin and medication event monitoring system (MEMS)] used simultaneously to evaluate naltrexone's efficacy and the impact of compliance on the size of observable treatment effects. One hundred and thirty-seven of 160 randomized alcoholic patients completed 12-weeks (84 days) of naltrexone or placebo and cognitive behavioral therapy (CBT) or motivational enhancement therapy (MET). Urine riboflavin was determined during study weeks 2, 6, and 12. The MEMS provided a detailed computerized record of when a participant opened their medication bottle throughout the trial. Baseline predictors of MEMS (80% openings) and urine riboflavin (>or=1,500 ng/mL by fluorimetry) compliance were examined. The effects of the treatments in the compliant participants defined by one, the other, or both methods were compared and contrasted with a previously reported intent-to-treat analysis where compliance was not taken into account. Age was predictive of compliance. 105 participants were deemed compliant via urine riboflavin criteria, 87 via MEMS, and 77 when both criteria were met, with no significant differences between treatment groups. The most compliant participants showed a significant medication by therapy interaction. Those treated with naltrexone/CBT showed more abstinence days (pcompliance measurement methods. Compliance measurement does appear to influence the evaluation of the efficacy of naltrexone within the context of CBT. Treatment effect sizes approximately doubled in the most compliant individuals. Measuring compliance by either of 2 distinct methods provides approximately similar results. As compliance with naltrexone

  20. Photo-induced fluorescence of magnesium derivatives of tetracycline antibiotics in wastewater samples

    International Nuclear Information System (INIS)

    Pena, A.; Albert-Garcia, J.R.; Silva, L.J.G.; Lino, C.M.; Calatayud, J. Martinez

    2010-01-01

    An analytical strategy, for the determination of tetracyclines (TCs), based on a HPLC system coupled with a photo-reactor followed by post-column derivatization was developed. Higher fluorescence emission after coupling the resulting photo-fragments with magnesium ions was observed for the determination of minocycline (MC), epitetracycline (ETC), tetracycline (TC) and doxycycline (DC). The manifold included a HPLC system with a photo-reactor (PTFE tubing helically coiled around a low-pressure mercury lamp), a mixing T-piece and a fluorescence detector. The derivatization reagent was delivered at 0.5 mL min -1 by a pump. After HPLC separation using a gradient system with a mobile phase containing oxalic acid 0.02 M and acetonitrile, TCs were irradiated for 60 s, and the resulting photo-fragments were mixed with the post-column derivatization reagent, and the magnesium derivatives of TCs were detected by fluorimetry (λ exc 386 nm, λ em 500 nm). The results obtained showed a significant increase of sensitivity due to photodegration of TCs, 45.4%, 37.6% and 25.3% for MC, TC and ETC respectively. For DC an increase of only 1.5% was observed. The developed method was successfully applied to TCs determination in hospital and municipal wastewater samples using solid phase extraction with Oasis HLB cartridges. The LOQs were 0.25, 0.15, 01 and 0.25 μg L -1 for TC, ETC, MC and DC, respectively. The recovery values oscillated between 107.1% and 92.4% for fortification of 2.5 μg L -1 of each antibiotic.

  1. Photo-induced fluorescence of magnesium derivatives of tetracycline antibiotics in wastewater samples

    Energy Technology Data Exchange (ETDEWEB)

    Pena, A., E-mail: apena@ff.uc.pt [Group of Health Surveillance, Center of Pharmaceutical Studies, Faculty of Pharmacy, University of Coimbra (Portugal); Albert-Garcia, J.R. [Department of Analytical Chemistry, Faculty of Chemistry, University of Valencia (Portugal); Silva, L.J.G.; Lino, C.M. [Group of Health Surveillance, Center of Pharmaceutical Studies, Faculty of Pharmacy, University of Coimbra (Portugal); Calatayud, J. Martinez [Department of Analytical Chemistry, Faculty of Chemistry, University of Valencia (Portugal)

    2010-07-15

    An analytical strategy, for the determination of tetracyclines (TCs), based on a HPLC system coupled with a photo-reactor followed by post-column derivatization was developed. Higher fluorescence emission after coupling the resulting photo-fragments with magnesium ions was observed for the determination of minocycline (MC), epitetracycline (ETC), tetracycline (TC) and doxycycline (DC). The manifold included a HPLC system with a photo-reactor (PTFE tubing helically coiled around a low-pressure mercury lamp), a mixing T-piece and a fluorescence detector. The derivatization reagent was delivered at 0.5 mL min{sup -1} by a pump. After HPLC separation using a gradient system with a mobile phase containing oxalic acid 0.02 M and acetonitrile, TCs were irradiated for 60 s, and the resulting photo-fragments were mixed with the post-column derivatization reagent, and the magnesium derivatives of TCs were detected by fluorimetry ({lambda}{sub exc} 386 nm, {lambda}{sub em} 500 nm). The results obtained showed a significant increase of sensitivity due to photodegration of TCs, 45.4%, 37.6% and 25.3% for MC, TC and ETC respectively. For DC an increase of only 1.5% was observed. The developed method was successfully applied to TCs determination in hospital and municipal wastewater samples using solid phase extraction with Oasis HLB cartridges. The LOQs were 0.25, 0.15, 01 and 0.25 {mu}g L{sup -1} for TC, ETC, MC and DC, respectively. The recovery values oscillated between 107.1% and 92.4% for fortification of 2.5 {mu}g L{sup -1} of each antibiotic.

  2. The chronic blockade of angiotensin I-converting enzyme eliminates the sex differences of serum cytokine levels of spontaneously hypertensive rats

    Energy Technology Data Exchange (ETDEWEB)

    Dalpiaz, P.L.M.; Lamas, A.Z.; Caliman, I.F. [Departamento de Ciências Fisiológicas, Universidade Federal do Espírito Santo, Vitória, ES (Brazil); Medeiros, A.R.S. [Ciências Biológicas e da Saúde, Instituto Federal do Espírito Santo, Vitória, ES (Brazil); Abreu, G.R.; Moysés, M.R. [Departamento de Ciências Fisiológicas, Universidade Federal do Espírito Santo, Vitória, ES (Brazil); Andrade, T.U. [Departamento de Farmácia, Centro Universitário de Vila Velha, Vila Velha, ES (Brazil); Alves, M.F.; Carmona, A.K. [Departamento de Biofísica, Escola Paulista de Medicina, Universidade Federal de São Paulo, São Paulo, SP (Brazil); Bissoli, N.S. [Departamento de Ciências Fisiológicas, Universidade Federal do Espírito Santo, Vitória, ES (Brazil)

    2013-02-01

    Sex hormones modulate the action of both cytokines and the renin-angiotensin system. However, the effects of angiotensin I-converting enzyme (ACE) on the proinflammatory and anti-inflammatory cytokine levels in male and female spontaneously hypertensive rats (SHR) are unclear. We determined the relationship between ACE activity, cytokine levels and sex differences in SHR. Female (F) and male (M) SHR were divided into 4 experimental groups each (n = 7): sham + vehicle (SV), sham + enalapril (10 mg/kg body weight by gavage), castrated + vehicle, and castrated + enalapril. Treatment began 21 days after castration and continued for 30 days. Serum cytokine levels (ELISA) and ACE activity (fluorimetry) were measured. Male rats exhibited a higher serum ACE activity than female rats. Castration reduced serum ACE in males but did not affect it in females. Enalapril reduced serum ACE in all groups. IL-10 (FSV = 16.4 ± 1.1 pg/mL; MSV = 12.8 ± 1.2 pg/mL), TNF-α (FSV = 16.6 ± 1.2 pg/mL; MSV = 12.8 ± 1 pg/mL) and IL-6 (FSV = 10.3 ± 0.2 pg/mL; MSV = 7.2 ± 0.2 pg/mL) levels were higher in females than in males. Ovariectomy reduced all cytokine levels and orchiectomy reduced IL-6 but increased IL-10 concentrations in males. Castration eliminated the differences in all inflammatory cytokine levels (IL-6 and TNF-α) between males and females. Enalapril increased IL-10 in all groups and reduced IL-6 in SV rats. In conclusion, serum ACE inhibition by enalapril eliminated the sexual dimorphisms of cytokine levels in SV animals, which suggests that enalapril exerts systemic anti-inflammatory and anti-hypertensive effects.

  3. Myricetin is a novel inhibitor of human inosine 5′-monophosphate dehydrogenase with anti-leukemia activity

    International Nuclear Information System (INIS)

    Pan, Huiling; Hu, Qian; Wang, Jingyuan; Liu, Zehui; Wu, Dang; Lu, Weiqiang; Huang, Jin

    2016-01-01

    Human inosine 5′-monophosphate dehydrogenase (hIMPDH) is a rate-limiting enzyme in the de novo biosynthetic pathway of purine nucleotides, playing crucial roles in cellular proliferation, differentiation, and transformation. Dysregulation of hIMPDH expression and activity have been found in a variety of human cancers including leukemia. In this study, we found that myricetin, a naturally occurring phytochemical existed in berries, wine and tea, was a novel inhibitor of human type 1 and type 2 IMPDH (hIMPDH1/2) with IC_5_0 values of 6.98 ± 0.22 μM and 4.10 ± 0.14 μM, respectively. Enzyme kinetic analysis using Lineweaver-Burk plot revealed that myricetin is a mix-type inhibitor for hIMPDH1/2. Differential scanning fluorimetry and molecular docking simulation data demonstrate that myricetin is capable of binding with hIMPDH1/2. Myricetin treatment exerts potent anti-proliferative and pro-apoptotic effects on K562 human leukemia cells in a dose-dependent manner. Importantly, cytotoxicity of myricetin on K562 cells were markedly attenuated by exogenous addition of guanosine, a salvage pathway of maintaining intracellular pool of guanine nucleotides. Taking together, these results indicate that natural product myricetin exhibits potent anti-leukemia activity by interfering with purine nucleotides biosynthetic pathway through the suppression of hIMPDH1/2 catalytic activity. - Highlights: • Myricetin, a common dietary flavonoid, is a novel inhibitor of hIMPDH1/2. • Myricetin directly binds with hIMPDH1/2 and induces cell cycle arrest and apoptosis of leukemia cells. • The cytotoxicity of myricetin on K562 cells is markedly attenuated by exogenous addition of guanosine.

  4. Increased MiR-221 expression in hepatocellular carcinoma tissues and its role in enhancing cell growth and inhibiting apoptosis in vitro

    International Nuclear Information System (INIS)

    Rong, Minhua; Chen, Gang; Dang, Yiwu

    2013-01-01

    MiR-221 is over-expressed in human hepatocellular carcinoma (HCC), but its clinical significance and function in HCC remains uncertain. The aim of the study was to investigate the relationship between miR-221 overexpression and clinicopathological parameters in HCC formalin-fixed paraffin-embedded (FFPE) tissues, and the effect of miR-221 inhibitor and mimic on different HCC cell lines in vitro. MiR-221 expression was detected using real time RT-qPCR in FFPE HCC and the adjacent noncancerous liver tissues. The relationship between miR-221 level and clinicopathological features was also analyzed. Furthermore, miR-221 inhibitor and mimic were transfected into HCC cell lines HepB3, HepG2 and SNU449. The effects of miR-221 on cell growth, cell cycle, caspase activity and apoptosis were also investigated by spectrophotometry, fluorimetry, fluorescence microscopy and flow cytometry, respectively. The relative expression of miR-221 in clinical TNM stages III and IV was significantly higher than that in the stages I and II. The miR-221 level was also upregulated in the metastatic group compared to the nonmetastatic group. Furthermore, miR-221 over-expression was related to the status of tumor capsular infiltration in HCC clinical samples. Functionally, cell growth was inhibited, cell cycle was arrested in G1/S-phase and apoptosis was increased by miR-221 inhibitor in vitro. Likewise, miR-221 mimic accelerated the cell growth. Expression of miR-221 in FFPE tissues could provide predictive significance for prognosis of HCC patients. Moreover, miR-221 inhibitor could be useful to suppress proliferation and induce apoptosis in HCC cells. Thus miR-221 might be a critical targeted therapy strategy for HCC

  5. [Sensitive Determination of Chondroitin Sulfate by Fluorescence Recovery of an Anionic Aluminum Phthalocyanine-Cationic Surfactant Ion-Association Complex Used as a Fluorescent Probe Emitting at Red Region].

    Science.gov (United States)

    Chen, Lin; Huang, Ping; Yang, Hui-qing; Deng, Ya-bin; Guo, Meng-lin; Li, Dong-hui

    2015-08-01

    Determination of chondroitin sulfate in the biomedical field has an important value. The conventional methods for the assay of chondroitin sulfate are still unsatisfactory in sensitivity, selectivity or simplicity. This work aimed at developing a novel method for sensitive and selective determination of chondroitin sulfate by fluorimetry. We found that some kinds of cationic surfactants have the ability to quench the fluorescence of tetrasulfonated aluminum phthalocyanine (AlS4Pc), a strongly fluorescent compound which emits at red region, with high efficiency. But, the fluorescence of the above-mentioned fluorescence quenching system recovered significantly when chondroitin sulfate (CS) exits. Tetradecyl dimethyl benzyl ammonium chloride(TDBAC) which was screened from all of the candidates of cationic surfactants was chosen as the quencher because it shows the most efficient quenching effect. It was found that the fluorescence of AlS4Pc was extremely quenched by TDBAC because of the formation of association complex between AlS4Pc and TDBAC. Fluorescence of the association complex recovered dramatically after the addition of chondroitin sulfate (CS) due to the ability of chondroitin sulfate to shift the association equilibrium of the association, leading to the release of AlS4Pc, thus resulting in an increase in the fluorescence of the reaction system. Based on this phenomenon, a novel method with simplicity, accuracy and sensitivity was developed for quantitative determination of CS. Factors including the reaction time, influencing factors and the effect of coexisting substances were investigated and discussed. Under optimum conditions the linear range of the calibration curve was 0.20~10.0 μg · mL(-1). The detection limit for CS was 0.070 μg · mL(-1). The method has been applied to the analysis of practical samples with satisfied results. This work expands the applications of AlS4Pc in biomedical area.

  6. Identification and Structure-Function Analysis of Subfamily Selective G Protein-Coupled Receptor Kinase Inhibitors

    Energy Technology Data Exchange (ETDEWEB)

    Homan, Kristoff T.; Larimore, Kelly M.; Elkins, Jonathan M.; Szklarz, Marta; Knapp, Stefan; Tesmer, John J.G. [Michigan; (Oxford)

    2015-02-13

    Selective inhibitors of individual subfamilies of G protein-coupled receptor kinases (GRKs) would serve as useful chemical probes as well as leads for therapeutic applications ranging from heart failure to Parkinson’s disease. To identify such inhibitors, differential scanning fluorimetry was used to screen a collection of known protein kinase inhibitors that could increase the melting points of the two most ubiquitously expressed GRKs: GRK2 and GRK5. Enzymatic assays on 14 of the most stabilizing hits revealed that three exhibit nanomolar potency of inhibition for individual GRKs, some of which exhibiting orders of magnitude selectivity. Most of the identified compounds can be clustered into two chemical classes: indazole/dihydropyrimidine-containing compounds that are selective for GRK2 and pyrrolopyrimidine-containing compounds that potently inhibit GRK1 and GRK5 but with more modest selectivity. The two most potent inhibitors representing each class, GSK180736A and GSK2163632A, were cocrystallized with GRK2 and GRK1, and their atomic structures were determined to 2.6 and 1.85 Å spacings, respectively. GSK180736A, developed as a Rho-associated, coiled-coil-containing protein kinase inhibitor, binds to GRK2 in a manner analogous to that of paroxetine, whereas GSK2163632A, developed as an insulin-like growth factor 1 receptor inhibitor, occupies a novel region of the GRK active site cleft that could likely be exploited to achieve more selectivity. However, neither compound inhibits GRKs more potently than their initial targets. This data provides the foundation for future efforts to rationally design even more potent and selective GRK inhibitors.

  7. Complexation modelling of uranium and other actinides by organic compounds of natural or synthetic origin

    International Nuclear Information System (INIS)

    Bouby, M.

    1998-01-01

    The future of nuclear wastes raises a lot of questions. Their resolution require an accurate knowledge of the physical, chemical and biological processes which affect the properties of radioelements constituting the wastes. 3 research themes have been approached. The experimental methods used are: neutronic activation analysis, UV-visible spectrophotometry and time-resolved induced laser spectro-fluorimetry. A part of the phenomena has been modelled by ionic strength correction models (as Davies or MSA). The main results have revealed: 1)the bio-sorption capacities of the microorganism (Mycobacterium phlei) for UO 2 2+ and NpO 2+ (in conditions where the specific adsorption capacities Qe(UO 2 2+ )=60 and Qe(NpO 2+ )=444 moles cations/g dry biomass 2)the retention capacities, in various leaching conditions, by this bacteria of the ions initially adsorbed 3)the complexation properties of 2 siderophores for the cations UO 2 2+ , U 4+ and Th 4+ . The thermodynamical equilibrium constants were determined for one of the siderophore: the pyoverdine A; they were such that KUO 2 2+ ≤KU 4+ ≤KTh 4+ 4)in very acidic media (HCl and HClO 4 until 12 M), the behaviour of the acylisoxazolone HPBI (1-phenyl-4-benzoyl-5-isoxazolone) and the value of its acidity thermodynamical constant is such that 0.13≤KATh≤0.32 at 25 degrees Celsius 5)the variations of the fluorescence properties of the uranyl cation in terms of the acidity of the concentrated media (HClO 4 and CF 3 SO 3 H) in which they are in solution; it seems that a complexation between the uranyl ion and the counter-ions present in solution occur. (O.M.)

  8. Glucose-stimulated calcium dynamics in islets of Langerhans in acute mouse pancreas tissue slices.

    Directory of Open Access Journals (Sweden)

    Andraž Stožer

    Full Text Available In endocrine cells within islets of Langerhans calcium ions couple cell stimulation to hormone secretion. Since the advent of modern fluorimetry, numerous in vitro studies employing primarily isolated mouse islets have investigated the effects of various secretagogues on cytoplasmic calcium, predominantly in insulin-secreting beta cells. Due to technical limitations, insights of these studies are inherently limited to a rather small subpopulation of outermost cells. The results also seem to depend on various factors, like culture conditions and duration, and are not always easily reconcilable with findings in vivo. The main controversies regard the types of calcium oscillations, presence of calcium waves, and the level of synchronized activity. Here, we set out to combine the in situ acute mouse pancreas tissue slice preparation with noninvasive fluorescent calcium labeling and subsequent confocal laser scanning microscopy to shed new light on the existing controversies utilizing an innovative approach enabling the characterization of responses in many cells from all layers of islets. Our experiments reproducibly showed stable fast calcium oscillations on a sustained plateau rather than slow oscillations as the predominant type of response in acute tissue slices, and that calcium waves are the mechanistic substrate for synchronization of oscillations. We also found indirect evidence that even a large amplitude calcium signal was not sufficient and that metabolic activation was necessary to ensure cell synchronization upon stimulation with glucose. Our novel method helped resolve existing controversies and showed the potential to help answer important physiological questions, making it one of the methods of choice for the foreseeable future.

  9. Myricetin is a novel inhibitor of human inosine 5′-monophosphate dehydrogenase with anti-leukemia activity

    Energy Technology Data Exchange (ETDEWEB)

    Pan, Huiling; Hu, Qian; Wang, Jingyuan; Liu, Zehui; Wu, Dang [Shanghai Key Laboratory of New Drug Design, School of Pharmacy, East China University of Science and Technology, 130 Mei Long Road, Shanghai 200237 (China); Lu, Weiqiang, E-mail: wqlu@bio.ecnu.edu.cn [Shanghai Key Laboratory of Regulatory Biology, Institute of Biomedical Sciences and School of Life Sciences, East China Normal University, 500 Dongchuan Road, Shanghai 200241 (China); Huang, Jin, E-mail: huangjin@ecust.edu.cn [Shanghai Key Laboratory of New Drug Design, School of Pharmacy, East China University of Science and Technology, 130 Mei Long Road, Shanghai 200237 (China)

    2016-09-02

    Human inosine 5′-monophosphate dehydrogenase (hIMPDH) is a rate-limiting enzyme in the de novo biosynthetic pathway of purine nucleotides, playing crucial roles in cellular proliferation, differentiation, and transformation. Dysregulation of hIMPDH expression and activity have been found in a variety of human cancers including leukemia. In this study, we found that myricetin, a naturally occurring phytochemical existed in berries, wine and tea, was a novel inhibitor of human type 1 and type 2 IMPDH (hIMPDH1/2) with IC{sub 50} values of 6.98 ± 0.22 μM and 4.10 ± 0.14 μM, respectively. Enzyme kinetic analysis using Lineweaver-Burk plot revealed that myricetin is a mix-type inhibitor for hIMPDH1/2. Differential scanning fluorimetry and molecular docking simulation data demonstrate that myricetin is capable of binding with hIMPDH1/2. Myricetin treatment exerts potent anti-proliferative and pro-apoptotic effects on K562 human leukemia cells in a dose-dependent manner. Importantly, cytotoxicity of myricetin on K562 cells were markedly attenuated by exogenous addition of guanosine, a salvage pathway of maintaining intracellular pool of guanine nucleotides. Taking together, these results indicate that natural product myricetin exhibits potent anti-leukemia activity by interfering with purine nucleotides biosynthetic pathway through the suppression of hIMPDH1/2 catalytic activity. - Highlights: • Myricetin, a common dietary flavonoid, is a novel inhibitor of hIMPDH1/2. • Myricetin directly binds with hIMPDH1/2 and induces cell cycle arrest and apoptosis of leukemia cells. • The cytotoxicity of myricetin on K562 cells is markedly attenuated by exogenous addition of guanosine.

  10. Arc is a flexible modular protein capable of reversible self-oligomerization

    Science.gov (United States)

    Myrum, Craig; Baumann, Anne; Bustad, Helene J.; Flydal, Marte Innselset; Mariaule, Vincent; Alvira, Sara; Cuéllar, Jorge; Haavik, Jan; Soulé, Jonathan; Valpuesta, José Maria; Márquez, José Antonio; Martinez, Aurora; Bramham, Clive R.

    2015-01-01

    The immediate early gene product Arc (activity-regulated cytoskeleton-associated protein) is posited as a master regulator of long-term synaptic plasticity and memory. However, the physicochemical and structural properties of Arc have not been elucidated. In the present study, we expressed and purified recombinant human Arc (hArc) and performed the first biochemical and biophysical analysis of hArc's structure and stability. Limited proteolysis assays and MS analysis indicate that hArc has two major domains on either side of a central more disordered linker region, consistent with in silico structure predictions. hArc's secondary structure was estimated using CD, and stability was analysed by CD-monitored thermal denaturation and differential scanning fluorimetry (DSF). Oligomerization states under different conditions were studied by dynamic light scattering (DLS) and visualized by AFM and EM. Biophysical analyses show that hArc is a modular protein with defined secondary structure and loose tertiary structure. hArc appears to be pyramid-shaped as a monomer and is capable of reversible self-association, forming large soluble oligomers. The N-terminal domain of hArc is highly basic, which may promote interaction with cytoskeletal structures or other polyanionic surfaces, whereas the C-terminal domain is acidic and stabilized by ionic conditions that promote oligomerization. Upon binding of presenilin-1 (PS1) peptide, hArc undergoes a large structural change. A non-synonymous genetic variant of hArc (V231G) showed properties similar to the wild-type (WT) protein. We conclude that hArc is a flexible multi-domain protein that exists in monomeric and oligomeric forms, compatible with a diverse, hub-like role in plasticity-related processes. PMID:25748042

  11. Fluorescent S-layer fusion proteins

    International Nuclear Information System (INIS)

    Kainz, B.

    2010-01-01

    This work describes the construction and characterisation of fluorescent S-layer fusion proteins used as building blocks for the fabrication of nanostructured monomolecular biocoatings on silica particles with defined fluorescence properties. The S-layer protein SgsE of Geobacillus stearothermophilus NRS 2004/3a was fused with the pH-dependant cyan, green and yellow variant of the green fluorescent protein (GFP) and the red fluorescent protein mRFP1. These fluorescent S-layer fusion proteins, acting as scaffold and optical sensing element simultaneously, were able to reassemble in solution and on silica particles forming 2D nanostructures with p2 lattice symmetry (a=11 ±0.5 nm, b=14 ±0.4 nm, g=80 ±1 o ). The pH-dependant fluorescence behaviour was studied with fluorimetry, confocal microscopy and flow cytometry. These fluorescent S-layer fusion proteins can be used as pH-sensor. 50% of the fluorescence intensity decreases at their calculated pKa values (pH6 - pH5). The fluorescence intensity of the GFP variants vanished completely between pH4 and pH3 whereas the chromophore of the red protein mRFP1 was only slightly affected in acidic conditions. At the isoelectric point of the S-layer coated silica particles (pH4.6 ±0.2) an increase in particle aggregation was detected by flow cytometry. The cyan and yellow fluorescent proteins were chosen to create a bi-fluorescent S-layer tandem fusion protein with the possibility for resonance energy transfer (FRET). A transfer efficiency of 20% and a molecular distance between the donor (ECFP) and acceptor (YFP) chromophores of around 6.2 nm could be shown. This bi-fluorescent ECFP-SgsE-YFP tandem fusion protein was able to reassemble on solid surfaces. The remarkable combination of fluorescence and self-assembly and the design of bi-functional S-layer tandem fusion protein matrices makes them to a promising tool in nanobiotechnology. (author) [de

  12. Macromolecular crowding gives rise to microviscosity, anomalous diffusion and accelerated actin polymerization.

    Science.gov (United States)

    Rashid, Rafi; Chee, Stella Min Ling; Raghunath, Michael; Wohland, Thorsten

    2015-04-30

    Macromolecular crowding (MMC) has been used in various in vitro experimental systems to mimic in vivo physiology. This is because the crowded cytoplasm of cells contains many different types of solutes dissolved in an aqueous medium. MMC in the extracellular microenvironment is involved in maintaining stem cells in their undifferentiated state (niche) as well as in aiding their differentiation after they have travelled to new locations outside the niche. MMC at physiologically relevant fractional volume occupancies (FVOs) significantly enhances the adipogenic differentiation of human bone marrow-derived mesenchymal stem cells during chemically induced adipogenesis. The mechanism by which MMC produces this enhancement is not entirely known. In the context of extracellular collagen deposition, we have recently reported the importance of optimizing the FVO while minimizing the bulk viscosity. Two opposing properties will determine the net rate of a biochemical reaction: the negative effect of bulk viscosity and the positive effect of the excluded volume, the latter being expressed by the FVO. In this study we have looked more closely at the effect of viscosity on reaction rates. We have used fluorimetry to measure the rate of actin polymerization and fluorescence correlation spectroscopy (FCS) to measure diffusion of various probes in solutions containing the crowder Ficoll at physiological concentrations. Similar to its effect on collagen, Ficoll enhanced the actin polymerization rate despite increasing the bulk viscosity. Our FCS measurements reveal a relatively minor component of anomalous diffusion. In addition, our measurements do suggest that microviscosity becomes relevant in a crowded environment. We ruled out bulk viscosity as a cause of the rate enhancement by performing the actin polymerization assay in glycerol. These opposite effects of Ficoll and glycerol led us to conclude that microviscosity becomes relevant at the length scale of the reacting

  13. Macromolecular crowding gives rise to microviscosity, anomalous diffusion and accelerated actin polymerization

    Science.gov (United States)

    Rashid, Rafi; Chee, Stella Min Ling; Raghunath, Michael; Wohland, Thorsten

    2015-05-01

    Macromolecular crowding (MMC) has been used in various in vitro experimental systems to mimic in vivo physiology. This is because the crowded cytoplasm of cells contains many different types of solutes dissolved in an aqueous medium. MMC in the extracellular microenvironment is involved in maintaining stem cells in their undifferentiated state (niche) as well as in aiding their differentiation after they have travelled to new locations outside the niche. MMC at physiologically relevant fractional volume occupancies (FVOs) significantly enhances the adipogenic differentiation of human bone marrow-derived mesenchymal stem cells during chemically induced adipogenesis. The mechanism by which MMC produces this enhancement is not entirely known. In the context of extracellular collagen deposition, we have recently reported the importance of optimizing the FVO while minimizing the bulk viscosity. Two opposing properties will determine the net rate of a biochemical reaction: the negative effect of bulk viscosity and the positive effect of the excluded volume, the latter being expressed by the FVO. In this study we have looked more closely at the effect of viscosity on reaction rates. We have used fluorimetry to measure the rate of actin polymerization and fluorescence correlation spectroscopy (FCS) to measure diffusion of various probes in solutions containing the crowder Ficoll at physiological concentrations. Similar to its effect on collagen, Ficoll enhanced the actin polymerization rate despite increasing the bulk viscosity. Our FCS measurements reveal a relatively minor component of anomalous diffusion. In addition, our measurements do suggest that microviscosity becomes relevant in a crowded environment. We ruled out bulk viscosity as a cause of the rate enhancement by performing the actin polymerization assay in glycerol. These opposite effects of Ficoll and glycerol led us to conclude that microviscosity becomes relevant at the length scale of the reacting

  14. Lactate up-regulates the expression of lactate oxidation complex-related genes in left ventricular cardiac tissue of rats.

    Directory of Open Access Journals (Sweden)

    Daniele Gabriel-Costa

    Full Text Available Besides its role as a fuel source in intermediary metabolism, lactate has been considered a signaling molecule modulating lactate-sensitive genes involved in the regulation of skeletal muscle metabolism. Even though the flux of lactate is significantly high in the heart, its role on regulation of cardiac genes regulating lactate oxidation has not been clarified yet. We tested the hypothesis that lactate would increase cardiac levels of reactive oxygen species and up-regulate the expression of genes related to lactate oxidation complex.Isolated hearts from male adult Wistar rats were perfused with control, lactate or acetate (20mM added Krebs-Henseleit solution during 120 min in modified Langendorff apparatus. Reactive oxygen species (O2●-/H2O2 levels, and NADH and NADPH oxidase activities (in enriched microsomal or plasmatic membranes, respectively were evaluated by fluorimetry while SOD and catalase activities were evaluated by spectrophotometry. mRNA levels of lactate oxidation complex and energetic enzymes MCT1, MCT4, HK, LDH, PDH, CS, PGC1α and COXIV were quantified by real time RT-PCR. Mitochondrial DNA levels were also evaluated. Hemodynamic parameters were acquired during the experiment. The key findings of this work were that lactate elevated cardiac NADH oxidase activity but not NADPH activity. This response was associated with increased cardiac O2●-/H2O2 levels and up-regulation of MCT1, MCT4, LDH and PGC1α with no changes in HK, PDH, CS, COXIV mRNA levels and mitochondrial DNA levels. Lactate increased NRF-2 nuclear expression and SOD activity probably as counter-regulatory responses to increased O2●-/H2O2.Our results provide evidence for lactate-induced up-regulation of lactate oxidation complex associated with increased NADH oxidase activity and cardiac O2●-/H2O2 driving to an anti-oxidant response. These results unveil lactate as an important signaling molecule regulating components of the lactate oxidation complex in

  15. Uranium and thorium based phosphate matrix: synthesis, characterizations and lixiviation; Matrices a base de phosphate d'uranium et de thorium: syntheses, caracterisations et lixiviation

    Energy Technology Data Exchange (ETDEWEB)

    Dacheux, N

    1995-03-01

    In the framework of the search for a ceramic material usable in the radioactive waste storage, uranium and thorium phosphates have been investigated. Their experimental synthesis conditions have been entirely reviewed, they lead to the preparation of four new compounds: U(UO{sub 2})(PO{sub 4}){sub 2}, U{sub 2}O(PO{sub 4}){sub 2}, UCIPO{sub 4}, 4H{sub 2}O, and Th{sub 4}(PO{sub 4}){sub 4}P{sub 2}O{sub 7}. Experimental evidenced are advanced for non existent compounds such as: U{sub 3}(PO{sub 4}){sub 4}, U{sub 2}O{sub 3}P{sub 2}O{sub 7} and Th{sub 3} (PO{sub 4}){sub 4}. Characterization by several techniques (X-rays and neutron powder diffractions, UV-Visible and Infra-red spectroscopies, XPS,...) were performed. The ab initio structure determination of U(UO{sub 2})(PO{sub 4}){sub 2} has been achieved by X-rays and refined by neutron diffractions. Through its physico-chemical analysis, we found that this compound was a new mixed valence uranium phosphate in which U{sup 4+} and UO{sub 2}{sup 2+} ions are ordered in pairs along parallel chains according to a new type of arrangement. Reaction mechanism, starting from UCIPO{sub 4}, 4H{sub 2}O and based on redox processes of uranium in solid state was set up. From two main matrices U(UO{sub 2})(PO{sub 4}){sub 2} and Th{sub 4}(PO{sub 4}){sub 4}P{sub 2}O{sub 7}, solid solutions were studied. They consist of replacement of U(IV) by Th(IV) and reversely. The leaching tests on pure, loaded and doped matrices were performed in terms of storage time, pH of solutions, and determined by the use of solids labelled with {sup 230}U or by the measurement of uranyl concentration by Laser-Induced Time-Resolved Spectro-fluorimetry. Average concentration of uranium in the liquid phase is around 10{sup -4} M to 10{sup -6} M. Taking into account the very low solubilities of the studied phosphate ceramics, we estimated their chemical performances promising as an answer to the important nuclear waste problem, if we compare them to the glasses

  16. Uranium and thorium based phosphate matrix: synthesis, characterizations and lixiviation

    International Nuclear Information System (INIS)

    Dacheux, N.

    1995-03-01

    In the framework of the search for a ceramic material usable in the radioactive waste storage, uranium and thorium phosphates have been investigated. Their experimental synthesis conditions have been entirely reviewed, they lead to the preparation of four new compounds: U(UO 2 )(PO 4 ) 2 , U 2 O(PO 4 ) 2 , UCIPO 4 , 4H 2 O, and Th 4 (PO 4 ) 4 P 2 O 7 . Experimental evidenced are advanced for non existent compounds such as: U 3 (PO 4 ) 4 , U 2 O 3 P 2 O 7 and Th 3 (PO 4 ) 4 . Characterization by several techniques (X-rays and neutron powder diffractions, UV-Visible and Infra-red spectroscopies, XPS,...) were performed. The ab initio structure determination of U(UO 2 )(PO 4 ) 2 has been achieved by X-rays and refined by neutron diffractions. Through its physico-chemical analysis, we found that this compound was a new mixed valence uranium phosphate in which U 4+ and UO 2 2+ ions are ordered in pairs along parallel chains according to a new type of arrangement. Reaction mechanism, starting from UCIPO 4 , 4H 2 O and based on redox processes of uranium in solid state was set up. From two main matrices U(UO 2 )(PO 4 ) 2 and Th 4 (PO 4 ) 4 P 2 O 7 , solid solutions were studied. They consist of replacement of U(IV) by Th(IV) and reversely. The leaching tests on pure, loaded and doped matrices were performed in terms of storage time, pH of solutions, and determined by the use of solids labelled with 230 U or by the measurement of uranyl concentration by Laser-Induced Time-Resolved Spectro-fluorimetry. Average concentration of uranium in the liquid phase is around 10 -4 M to 10 -6 M. Taking into account the very low solubilities of the studied phosphate ceramics, we estimated their chemical performances promising as an answer to the important nuclear waste problem, if we compare them to the glasses used at the present time. (author)

  17. The effects of 3,4-methylenedioxymethamphetamine (MDMA) on nicotinic receptors: Intracellular calcium increase, calpain/caspase 3 activation, and functional upregulation

    International Nuclear Information System (INIS)

    Garcia-Rates, Sara; Camarasa, Jordi; Sanchez-Garcia, Ana I.; Gandia, Luis; Escubedo, Elena; Pubill, David

    2010-01-01

    Previous work by our group demonstrated that homomeric α7 nicotinic acetylcholine receptors (nAChR) play a role in the neurotoxicity induced by 3,4-methylenedioxymethamphetamine (MDMA), as well as the binding affinity of this drug to these receptors. Here we studied the effect of MDMA on the activation of nAChR subtypes, the consequent calcium mobilization, and calpain/caspase 3 activation because prolonged Ca 2+ increase could contribute to cytotoxicity. As techniques, we used fluorimetry in Fluo-4-loaded PC12 cells and electrophysiology in Xenopus oocytes. MDMA produced a rapid and sustained increase in calcium without reaching the maximum effect induced by ACh. It also concentration-dependently inhibited the response induced by ACh, nicotine, and the specific α7 agonist PNU 282987 with IC 50 values in the low micromolar range. Similarly, MDMA induced inward currents in Xenopus oocytes transfected with human α7 but not with α4β2 nAChR and inhibited ACh-induced currents in both receptors in a concentration-dependent manner. The calcium response was inhibited by methyllycaconitine (MLA) and α-bungarotoxin but not by dihydro-β-erythroidine. These results therefore indicate that MDMA acts as a partial agonist on α7 nAChRs and as an antagonist on the heteromeric subtypes. Subsequently, calcium-induced Ca 2+ release from the endoplasmic reticulum and entry through voltage-operated calcium channels are also implicated as proved using specific antagonists. In addition, treatment with MDMA for 24 h significantly increased basal Ca 2+ levels and induced an increase in α-spectrin breakdown products, which indicates that calpain and caspase 3 were activated. These effects were inhibited by pretreatment with MLA. Moreover, pretreatment with MDMA induced functional upregulation of calcium responses to specific agonists of both heteromeric and α7 nAChR. Sustained calcium entry and calpain activation could favor the activation of Ca 2+ -dependent enzymes such as

  18. Norm survey in Argentina

    International Nuclear Information System (INIS)

    Canoba, Analia Cecilia; Gnoni, Gabriela Alejandra; Truppa, W.

    2008-01-01

    The oil and gas industry, which is especially significant in Argentina, is one industry that concentrates natural radionuclides during its processes. In addition, there are a few underground mines under development, where radon levels may be high. The Nuclear Regulatory Authority of Argentina (ARN) carried out a project with the objective of evaluating NORM, mainly in these types of industries. Eight facilities were characterized, three related to the gas industry, four related to the oil industry and a survey of radon gas in a gold underground mine. First, background measurements were made and then a screening survey was carried out to detect values above background. Of the values obtained, 57% were in the background range, 19% were below 2 μSv/h, 15% were in the range 2-10 μSv/h and 9% were above 10 μSv/h. Some values were as high as 400 μSv/h. The annual effective doses were estimated to be in the range 0.02-1.6 mSv/a. Samples were taken and later analysed by gamma spectrometry, liquid scintillation and fluorimetry. It was confirmed that the main radionuclides involved in the oil and gas extraction process are 226 Ra and 228 Ra. The radium isotope concentrations measured in some samples were above the exemption values established by the International Basic Safety Standards. Elevated radon levels were detected in gas facilities and in the gold mine under development. The values obtained in gas facilities showed that radon concentrates in the ethane and propane flows. As the flows in the gas industry are confined, it does not mean an exposure during normal operation. In the case of the gold mine, the values detected were informed to the pertinent authorities as well as the facility in order to take actions to reduce concentrations below the action levels. Finally, protective measures to reduce occupational doses in the cleaning and maintenance processes were suggested, as well as for storage of NORM-contaminated items. (author)

  19. Determination of uranium in ground water using different analytical techniques

    International Nuclear Information System (INIS)

    Sahu, S.K.; Maity, Sukanta; Bhangare, R.C.; Pandit, G.G.; Sharma, D.N.

    2014-10-01

    collected from different locations across India using three analytical techniques; fluorimetry, inductively coupled plasma atomic emission spectrometry (ICP-AES) and differential pulse adsorptive stripping voltammetry (DPASV). The performance, response, sensitivity, reproducibility, linear range of these three methodologies were compared. It was found that the voltammetric technique was the most sensitive, efficient and economic process for trace level analyses. (author)

  20. Radionuclide concentration in drinking water from public supplies in the metropolitan area of Recife, Brazil

    International Nuclear Information System (INIS)

    Melo, N.P.; Amaral, R.S.; Silva, C.M.; Hazin, C.A.; Lima, R.A.; Valentim, E.

    2000-01-01

    There has been an increasing concern with the assessment of human exposure to naturally occurring radionuclides as they account for around 70% of the total dose received by the population. Most of the exposure derived from natural terrestrial sources comes from the radionuclides in the uranium series, which is headed by the long-lived isotope 238 U. Radionuclides pertaining to this decay chain, such as, 226 Ra, 222 Ra, 210 Pb and 210 Po are the most important to be studied, as far as the dose to the population is concerned. In order to verify if the levels of radioactive contaminants in the water from public supplies are in compliance with the limits set by the Ministry of Health, a survey has been carried out in the metropolitan area of Recife, Brazil. This paper reports the results of this survey for both, superficial and groundwater supplies. Water samples collected from 107 sampling points were analyzed to determine their content of natural uranium, 226 Ra, 222 Ra, 210 Pb and 210 Po. Uranium concentrations were determined by fluorimetry after extraction with an organic solvent and deposition in LiF/NaF pellets, which were fused at 1000degC. Radium-226 was determined by the emanation technique, whereas 222 Rn was determined by liquid scintillation counting. Polonium-210, on the other hand, was determined by alpha spectrometry, following its spontaneous plating in copper disks immersed in the water samples. Measured concentrations ranged from 43 to 1330 mBq/L for natural uranium, 5.3 to 83.7 Bq/L for 222 Rn and from below the MDL (22 mBq/L) to 337 mBq/L for 210 Po. The annual intake of each radionuclide was calculated by considering an ingestion of 2L of water per day. The dose to each organ considered important to the ingestion pathway was calculated by using the masses given in the ICRP 23 for the standard man and by applying the appropriate dose conversion factors. (author)

  1. Mechanisms of tramadol-related neurotoxicity in the rat: Does diazepam/tramadol combination play a worsening role in overdose?

    Energy Technology Data Exchange (ETDEWEB)

    Lagard, Camille, E-mail: camille.lagard@gmail.com [Inserm, U1144, Paris (France); UMR-S 1144, Paris-Descartes University, Paris (France); UMR-S 1144, Paris-Diderot University, Paris (France); Chevillard, Lucie, E-mail: luciechevillard@gmail.com [Inserm, U1144, Paris (France); UMR-S 1144, Paris-Descartes University, Paris (France); UMR-S 1144, Paris-Diderot University, Paris (France); Malissin, Isabelle, E-mail: isabellemalissin@gmail.com [Assistance Publique – Hôpitaux de Paris, Lariboisière Hospital, Department of Medical and Toxicological Critical Care, Paris (France); Risède, Patricia, E-mail: patricia.risede@inserm.fr [Inserm, U1144, Paris (France); UMR-S 1144, Paris-Descartes University, Paris (France); UMR-S 1144, Paris-Diderot University, Paris (France); Callebert, Jacques, E-mail: jacques.callebert@aphp.fr [Inserm, U1144, Paris (France); UMR-S 1144, Paris-Descartes University, Paris (France); UMR-S 1144, Paris-Diderot University, Paris (France); Assistance Publique – Hôpitaux de Paris, Lariboisière Hospital, Laboratory of Biochemistry and Molecular Biology, Paris (France); Labat, Laurence, E-mail: laurence.labat@aphp.fr [Inserm, U1144, Paris (France); UMR-S 1144, Paris-Descartes University, Paris (France); UMR-S 1144, Paris-Diderot University, Paris (France); Assistance Publique – Hôpitaux de Paris, Cochin Hospital, Laboratory of Toxicology, Paris (France); Launay, Jean-Marie, E-mail: jean-marie.launay@aphp.fr [Assistance Publique – Hôpitaux de Paris, Lariboisière Hospital, Laboratory of Biochemistry and Molecular Biology, Paris (France); Inserm, U942, Paris (France); and others

    2016-11-01

    Poisoning with opioid analgesics including tramadol represents a challenge. Tramadol may induce respiratory depression, seizures and serotonin syndrome, possibly worsened when in combination to benzodiazepines. Our objectives were to investigate tramadol-related neurotoxicity, consequences of diazepam/tramadol combination, and mechanisms of drug-drug interactions in rats. Median lethal-doses were determined using Dixon–Bruce's up-and-down method. Sedation, seizures, electroencephalography and plethysmography parameters were studied. Concentrations of tramadol and its metabolites were measured using liquid-chromatography-high-resolution-mass-spectrometry. Plasma, platelet and brain monoamines were measured using liquid-chromatography coupled to fluorimetry. Median lethal-doses of tramadol and diazepam/tramadol combination did not significantly differ, although time-to-death was longer with combination (P = 0.04). Tramadol induced dose-dependent sedation (P < 0.05), early-onset seizures (P < 0.001) and increase in inspiratory (P < 0.01) and expiratory times (P < 0.05). The diazepam/tramadol combination abolished seizures but significantly enhanced sedation (P < 0.01) and respiratory depression (P < 0.05) by reducing tidal volume (P < 0.05) in addition to tramadol-related increase in respiratory times, suggesting a pharmacodynamic mechanism of interaction. Plasma M1 and M5 metabolites were mildly increased, contributing additionally to tramadol-related respiratory depression. Tramadol-induced early-onset increase in brain concentrations of serotonin and norepinephrine was not significantly altered by the diazepam/tramadol combination. Interestingly neither pretreatment with cyproheptadine (a serotonin-receptor antagonist) nor a benserazide/5-hydroxytryptophane combination (enhancing brain serotonin) reduced tramadol-induced seizures. Our study shows that diazepam/tramadol combination does not worsen tramadol-induced fatality risk but alters its toxicity pattern

  2. Plant HDAC inhibitor chrysin arrest cell growth and induce p21WAF1 by altering chromatin of STAT response element in A375 cells

    International Nuclear Information System (INIS)

    Pal-Bhadra, Manika; Bhadra, Utpal; Ramaiah, M Janaki; Reddy, T Lakshminarayan; Krishnan, Anita; Pushpavalli, SNCVL; Babu, K Suresh; Tiwari, Ashok K; Rao, J Madhusudana; Yadav, Jhillu S

    2012-01-01

    Chrysin and its analogues, belongs to flavonoid family and possess potential anti-tumour activity. The aim of this study is to determine the molecular mechanism by which chrysin controls cell growth and induce apoptosis in A375 cells. Effect of chrysin and its analogues on cell viability and cell cycle analysis was determined by MTT assay and flowcytometry. A series of Western blots was performed to determine the effect of chrysin on important cell cycle regulatory proteins (Cdk2, cyclin D1, p53, p21, p27). The fluorimetry and calorimetry based assays was conducted for characterization of chrysin as HDAC inhibitor. The changes in histone tail modification such as acetylation and methylation was studied after chrysin treatment was estimated by immuno-fluorescence and western blot analysis. The expression of Bcl-xL, survivin and caspase-3 was estimated in chrysin treated cells. The effect of chrysin on p21 promoter activity was studied by luciferase and ChIP assays. Chrysin cause G1 cell cycle arrest and found to inhibit HDAC-2 and HDAC-8. Chrysin treated cells have shown increase in the levels of H3acK14, H4acK12, H4acK16 and decrease in H3me2K9 methylation. The p21 induction by chrysin treatment was found to be independent of p53 status. The chromatin remodelling at p21 WAF1 promoter induces p21 activity, increased STAT-1 expression and epigenetic modifications that are responsible for ultimate cell cycle arrest and apoptosis. Chrysin shows in vitro anti-cancer activity that is correlated with induction of histone hyperacetylation and possible recruitment of STAT-1, 3, 5 proteins at STAT (−692 to −684) region of p21 promoter. Our results also support an unexpected action of chrysin on the chromatin organization of p21 WAF1 promoter through histone methylation and hyper-acetylation. It proposes previously unknown sequence specific chromatin modulations in the STAT responsive elements for regulating cell cycle progression negatively via the induction of the CDK

  3. SPECTROPHOTOMETRIC DETERMINATION OF ACETYLCYSTEINE IN PHARMACEUTICAL FORMULATIONS USING 2,3-DICHLORO-1,4-NAPTHOQUINONE

    Directory of Open Access Journals (Sweden)

    A. O. Donchenko

    2015-04-01

    Full Text Available The aim of research was the development and validation ofspectrophotometric method foracetylcysteine assay in pharmaceutical formulations.Тhe proposed method is based on the reaction with 2,3-dichloro-1,4-naphthoquinone and the formation of colored products that exhibit absorption maxima at 425 nm. Introduction Many analytical methods have been published for acetylcysteine assay in pharmaceutical formulations as high-performance liquid chromatography (HPLC, fluorimetry and chemiluminescence. Some of these methods are time consuming or require expensive equipment. Other published methods suffer from lack of selectivity and sensitivity. Spectrophotometry is the most widely used technique in pharmaceutical analysis because it is simple, economic, and easily available to most quality control laboratories. In the present work, we propose a simple and accurate spectrophotometric method for acetylcysteine assay in pharmaceutical formulations. Materials and Methods Reagents: Reference standard acetylcysteinesubstance; 2,3-dichloro-1,4-naphthoquinone. All chemicals and solvents were of analytical grade. DMFA was used as a solvent. Pharmaceutical preparations:powder for oral solution «ACC 200» 200 mgseries number50026151 (Salutas Pharma CmbH, Germany; effervescent tablets «Fluimucil» 600 mg (Zambon S.P.A., Italy and «ACC LONG» 600 mg (Salutas Pharma CmbH, Germany series numbers 321284 and DH2740; solution for injections «Fluimucil» 100 mg/ml (Zambon S.P.A., Italyseries number28002492. Solutions: Acetylcysteine stock solution (0,16%; DMFAsolution of 2,3-dichloro-1,4-naphthoquinone (4%. Equipment Analytical balance (ABT-120-5DM; UV-VIS spectrophotometer (Specord 200; water bath (MemmertWNB 7-45;quartz cells. Results Acetylcysteine was determined using a spectrophotometric method based on the reaction with 2,3-dichloro-1,4-naphthoquinone to form yellow colored reaction products with absorption maxima at 425 nm. The effect of reaction time and

  4. Measurements of Tenorm in an enterprise related with the petroleum industry; Mediciones de Tenorm en una empresa relacionada con la industria petrolera

    Energy Technology Data Exchange (ETDEWEB)

    Gnoni, G.; Canoba, A. [Autoridad Regulatoria Nuclear, Av. Del Libertador 8250 Ciudad de Buenos Aires (1429) (Argentina)]. e-mail: ggnoni@cae.arn.gov.ar

    2006-07-01

    Some industries in its processes concentrate natural radionuclides. Among these industries are the industry of the gas and the petroleum. These materials naturally radioactive concentrated by certain industries are known as TENORM (technologically enhanced naturally occurring radioactive material). As part of a project that has by objective to determine possible contaminations with TENORM materials in the industry, it was carried out an evaluation to a company related with the oil industry. The company provides teams and systems used in the production wells. In the plant it proceeds to the armed one with new components or with recovered parts of used equipment. Its were carried out measurements of dose rate in the place. In the first place it was carried out a screening monitoring, in order to detect hot points. In those cases in that hot points were not detected, pieces were selected to measure in contact, considering it origin, function and visual inspection of the same ones. The measured dose rate in most of the pieces was inside the radiation bottom levels. Its were found some pieces with greater dose rates at the bottom and only isolated cases that were up to ten times greater to the same one. The annual effective dose estimated in preservative form, only considering the external exposure via, starting from the most high value measured was similar to 0.6 mSv.a{sup -1}, value far from the one dose limit settled down for workers (20 mSv.a{sup -1}) and also smaller to the one dose limit settled down for public (1 mSv.a{sup -1}). Its took oily crusts samples, wash water, and cleaning sands for the later analysis in the laboratory. The samples were analyzed by gamma spectrometry and the uranium concentration by fluorimetry. Starting from the measurements carried out in the laboratories, it was confirmed that the involved radionuclides are the Ra-226 and Ra-228 and that the uranium doesn't concentrate on the present inlays in the pieces analyzed, in agreement

  5. Radionuclide Activity Concentrations in Spas of Argentina

    International Nuclear Information System (INIS)

    Gnoni, G.; Czerniczyniec, M.; Palacios, M.

    2011-01-01

    Geothermal waters have been used on a large scale for bathing, drinking and medical purposes. These waters often have a very high mineral content because solubility increases with temperature. Ground waters are in close contact with soil and rocks containing radium. Once formed by decay from radium, radon gas (Rn-222) may diffuse through rocks pores and geological discontinuities and may dissolve in these waters. Radon and other natural radionuclides are transported to the surface where radon can easily diffuse into the atmosphere. Then it may be possible to find out significant radon levels at places like geothermal spas. In this work the most important natural radionuclide activity concentrations in different thermal spas of Argentina were measured to characterize waters and to evaluate the exposure of workers and members of the public. Three passive methods were used to measure radon in air. One of them is an screening method based on the radon adsorption on activated charcoal. The other two methods are time integrated ones, CR-39 or Makrofol tracks detectors, which can be exposed between two and three months. To characterize waters other natural radionuclides have been also measured. Uranium concentration was measured by fluorimetry. Ra-226 and Pb-210 measurements were performed by radiochemical methods and liquid scintillation. The results obtained were compared with the guidelines values recommended by WHO and EPA for drinking waters and, in the case of radon in air, the results were compared with values established by BSS-115. In order to assess worker doses, the higher value measured for radon in air and real scenario data were taken into account. Moreover, in situ dose rate measurements were also performed and then compared with natural background values. In relation with water characterization, almost all values obtained for the geothermal waters analyzed were below the corresponding guidance levels. Taking into account the highest value measured of radon

  6. Reduction in photosynthetic efficiency of Cladophora glomerata, induced by overlying canopies of Lemna spp.

    Science.gov (United States)

    Parr, L B; Perkins, R G; Mason, C F

    2002-04-01

    The duckweeds Lemna minor L. and L. minuscula Herter reduced PSII quantum efficiency (F'q/F'm) of the filamentous green alga Cladophora glomerata Kützing by up to 42% over seven days when floating above mats of C. glomerata in containers. Dissolved oxygen (DO) increased by 23% at 30 degrees C in containers with C. glomerata over controls. But when the water surface in the containers was covered with Lemna spp. floating above C. glomerata, DO was 83% lower at 30 degrees C over seven days than in control samples with no duckweed or alga. Dissolved oxygen was lower beneath a thick mat (1 cm) of either Lemna spp. covering the surface than under a thin layer (single-frond canopy). PAM fluorimetry showed that maximum PSII efficiency (Fv/Fm) of C. glomerata in containers was reduced under a canopy of L. minor by 17% over seven days, and under L. minuscula by 22%. F'q/F'm of C. glomerata in containers exposed to 51 micromol m(-2) s(-1) PPFD decreased under a canopy of L. minor by 16% over seven days, and under L. minuscula by 19% compared to controls. When light response curves were compared, F'q/F'm was significantly reduced under canopies of L. minor at the highest temperatures tested (28 degrees C and 30 degrees C). L. minor significantly reduced relative electron transport rate (rel. ETR) of the controls by up to 71% at 30 degrees C. Relative electron transport rate did not reach light saturation point (Esat) except at 28 degrees and 30 degrees C under mats of L. minor. Whereas the highest rate of production (rel. ETRmax) and Esat increased with temperature in controls, under a canopy of Lemna, decreases were observed. It is suggested that, during periods of high summer temperature and irradiance, shading inhibits oxygenic photosynthesis in mats of C. glomerata beneath canopies of Lenma spp. This results in less oxygen being produced by the C. glomerata (oxygen produced by Lemna spp. is not released into the water), and this may further inhibit the C. glomerata by

  7. Mutants of Yarrowia lipolytica NCIM 3589 grown on waste cooking oil as a biofactory for biodiesel production.

    Science.gov (United States)

    Katre, Gouri; Ajmera, Namasvi; Zinjarde, Smita; RaviKumar, Ameeta

    2017-10-24

    Oleaginous yeasts are fast emerging as a possible feedstock for biodiesel production. Yarrowia lipolytica, a model oleaginous yeast is known to utilize a variety of hydrophobic substrates for lipid accumulation including waste cooking oil (WCO). Approaches to increase lipid content in this yeast include metabolic engineering which requires manipulation of multiple genes in the lipid biosynthesis pathway. A classical and cost-effective approach, namely, random chemical mutagenesis on the yeast can lead to increased production of biodiesel as is explored here. In this study, chemical mutagenesis using the alkylating agent, N- methyl-N'-nitro-N-nitrosoguanidine (MNNG) as well as an additional treatment with cerulenin, a fatty acid synthase inhibitor generated 800 mutants of Y. lipolytica NCIM 3589 (761 MNNG treated and 39 MNNG + cerulenin treated). A three-stage screening using Sudan Black B plate technique, Nile red fluorimetry and total lipid extraction using solvent was performed, which enabled selection of ten high lipid yielding mutants. Time course studies of all the ten mutants were further undertaken in terms of biomass, lipid yield and lipid content to select three stable mutants (YlB6, YlC7 and YlE1) capable of growing and accumulating lipid on WCO, with lipid contents of 55, 60 and 67% as compared to 45% for the wild type. The mutants demonstrated increased volumetric lipid productivities (0.062, 0.044 and 0.041 g L -1  h -1 ) as compared to the wild type (0.033 g L -1  h -1 ). The fatty acid profile of the three mutants consisted of a high content of C16 and C18 saturated and monounsaturated fatty acids and was found to be suitable for biodiesel production. The fuel properties, namely, density, kinematic viscosity, total acid number, iodine value of the three mutants were evaluated and found to lie within the limits specified by internationally accepted standards. Additionally, it was noted that the mutants demonstrated better cetane numbers and

  8. Mechanisms of tramadol-related neurotoxicity in the rat: Does diazepam/tramadol combination play a worsening role in overdose?

    International Nuclear Information System (INIS)

    Lagard, Camille; Chevillard, Lucie; Malissin, Isabelle; Risède, Patricia; Callebert, Jacques; Labat, Laurence; Launay, Jean-Marie

    2016-01-01

    Poisoning with opioid analgesics including tramadol represents a challenge. Tramadol may induce respiratory depression, seizures and serotonin syndrome, possibly worsened when in combination to benzodiazepines. Our objectives were to investigate tramadol-related neurotoxicity, consequences of diazepam/tramadol combination, and mechanisms of drug-drug interactions in rats. Median lethal-doses were determined using Dixon–Bruce's up-and-down method. Sedation, seizures, electroencephalography and plethysmography parameters were studied. Concentrations of tramadol and its metabolites were measured using liquid-chromatography-high-resolution-mass-spectrometry. Plasma, platelet and brain monoamines were measured using liquid-chromatography coupled to fluorimetry. Median lethal-doses of tramadol and diazepam/tramadol combination did not significantly differ, although time-to-death was longer with combination (P = 0.04). Tramadol induced dose-dependent sedation (P < 0.05), early-onset seizures (P < 0.001) and increase in inspiratory (P < 0.01) and expiratory times (P < 0.05). The diazepam/tramadol combination abolished seizures but significantly enhanced sedation (P < 0.01) and respiratory depression (P < 0.05) by reducing tidal volume (P < 0.05) in addition to tramadol-related increase in respiratory times, suggesting a pharmacodynamic mechanism of interaction. Plasma M1 and M5 metabolites were mildly increased, contributing additionally to tramadol-related respiratory depression. Tramadol-induced early-onset increase in brain concentrations of serotonin and norepinephrine was not significantly altered by the diazepam/tramadol combination. Interestingly neither pretreatment with cyproheptadine (a serotonin-receptor antagonist) nor a benserazide/5-hydroxytryptophane combination (enhancing brain serotonin) reduced tramadol-induced seizures. Our study shows that diazepam/tramadol combination does not worsen tramadol-induced fatality risk but alters its toxicity pattern

  9. Measurements of Tenorm in an enterprise related with the petroleum industry

    International Nuclear Information System (INIS)

    Gnoni, G.; Canoba, A.

    2006-01-01

    Some industries in its processes concentrate natural radionuclides. Among these industries are the industry of the gas and the petroleum. These materials naturally radioactive concentrated by certain industries are known as TENORM (technologically enhanced naturally occurring radioactive material). As part of a project that has by objective to determine possible contaminations with TENORM materials in the industry, it was carried out an evaluation to a company related with the oil industry. The company provides teams and systems used in the production wells. In the plant it proceeds to the armed one with new components or with recovered parts of used equipment. Its were carried out measurements of dose rate in the place. In the first place it was carried out a screening monitoring, in order to detect hot points. In those cases in that hot points were not detected, pieces were selected to measure in contact, considering it origin, function and visual inspection of the same ones. The measured dose rate in most of the pieces was inside the radiation bottom levels. Its were found some pieces with greater dose rates at the bottom and only isolated cases that were up to ten times greater to the same one. The annual effective dose estimated in preservative form, only considering the external exposure via, starting from the most high value measured was similar to 0.6 mSv.a -1 , value far from the one dose limit settled down for workers (20 mSv.a -1 ) and also smaller to the one dose limit settled down for public (1 mSv.a -1 ). Its took oily crusts samples, wash water, and cleaning sands for the later analysis in the laboratory. The samples were analyzed by gamma spectrometry and the uranium concentration by fluorimetry. Starting from the measurements carried out in the laboratories, it was confirmed that the involved radionuclides are the Ra-226 and Ra-228 and that the uranium doesn't concentrate on the present inlays in the pieces analyzed, in agreement with the fact that

  10. Uranium and thorium based phosphate matrix: synthesis, characterizations and lixiviation; Matrices a base de phosphate d'uranium et de thorium: syntheses, caracterisations et lixiviation

    Energy Technology Data Exchange (ETDEWEB)

    Dacheux, N

    1995-03-01

    In the framework of the search for a ceramic material usable in the radioactive waste storage, uranium and thorium phosphates have been investigated. Their experimental synthesis conditions have been entirely reviewed, they lead to the preparation of four new compounds: U(UO{sub 2})(PO{sub 4}){sub 2}, U{sub 2}O(PO{sub 4}){sub 2}, UCIPO{sub 4}, 4H{sub 2}O, and Th{sub 4}(PO{sub 4}){sub 4}P{sub 2}O{sub 7}. Experimental evidenced are advanced for non existent compounds such as: U{sub 3}(PO{sub 4}){sub 4}, U{sub 2}O{sub 3}P{sub 2}O{sub 7} and Th{sub 3} (PO{sub 4}){sub 4}. Characterization by several techniques (X-rays and neutron powder diffractions, UV-Visible and Infra-red spectroscopies, XPS,...) were performed. The ab initio structure determination of U(UO{sub 2})(PO{sub 4}){sub 2} has been achieved by X-rays and refined by neutron diffractions. Through its physico-chemical analysis, we found that this compound was a new mixed valence uranium phosphate in which U{sup 4+} and UO{sub 2}{sup 2+} ions are ordered in pairs along parallel chains according to a new type of arrangement. Reaction mechanism, starting from UCIPO{sub 4}, 4H{sub 2}O and based on redox processes of uranium in solid state was set up. From two main matrices U(UO{sub 2})(PO{sub 4}){sub 2} and Th{sub 4}(PO{sub 4}){sub 4}P{sub 2}O{sub 7}, solid solutions were studied. They consist of replacement of U(IV) by Th(IV) and reversely. The leaching tests on pure, loaded and doped matrices were performed in terms of storage time, pH of solutions, and determined by the use of solids labelled with {sup 230}U or by the measurement of uranyl concentration by Laser-Induced Time-Resolved Spectro-fluorimetry. Average concentration of uranium in the liquid phase is around 10{sup -4} M to 10{sup -6} M. Taking into account the very low solubilities of the studied phosphate ceramics, we estimated their chemical performances promising as an answer to the important nuclear waste problem, if we compare them to the glasses

  11. Colloid and radionuclide retention mechanisms in fractured rock under near-natural flow conditions

    International Nuclear Information System (INIS)

    Delos, A.; Schaefer, T.; Geckeis, H.; Guimera, J.; Carrera, J.; Fanghaenel, T.

    2005-01-01

    Full text of publication follows: Experiments in fractured host rock (Grimsel Test Site, GTS, Switzerland) revealed that the colloid relevance for actinide migration is high due to the specific geochemical groundwater conditions [1]. However, even under such conditions it is found that retention of colloids and colloid-borne actinides becomes significant under near-natural groundwater flow rates (1-10 m/a) [2]. Underlying mechanisms of colloid and radionuclide retention are not well understood up to now. The present study co-funded by the NoE ACTINET-6 focuses on (i) the kinetics of actinide-colloid interactions and (ii) the relevance of matrix diffusion as a competition process to other retention mechanisms which affect the actinides behavior in fractured rock systems such as the Grimsel granodiorite. Colloid migration is studied with well defined model colloids as e.g. fluorescence dyed carboxylated polystyrene particles, and natural colloids extracted from bentonite (FEBEX) and from fracture filling material (GTS). In order to study the influence of matrix porosity on actinides migration, those experiments are performed in columns of well defined geometry filled with microporous unmodified silica spheres, porous ceramic material and natural fracture filling material from the GTS. The behaviour of actinides (Pu(IV) and Am(III)) sorbed onto bentonite colloids is investigated in column and batch experiments. All experiments are performed under anoxic conditions. Colloid characterization methods used in this study include the combination of photon correlation spectroscopy (PCS), laser-induced breakdown detection (LIBD), fluorimetry and field flow fractionation (FFF). Experimental results and their application to the parametrisation of reactive colloid transport models are discussed. [1] Geckeis H, Schaefer T, Hauser W, Rabung T, Missana T, Degueldre C, Moeri A, Eikenberg J, Fierz T, Alexander WR (2004) Results of the Colloid and Radionuclide Retention experiment

  12. Estudo in vivo de atividade anti-radicalar por quantificação de peróxidos cutâneos In vivo antiradicalar activity by skin peroxidies quantification

    Directory of Open Access Journals (Sweden)

    Rodrigo Fuscelli Pytel

    2005-12-01

    efficacy of a new high-activity antioxidant complex (tocopheryl acetate, licopene and a pool of chlorogenic acids rich in cafeic acid. METHODS: In this non-invasive and placebo controlled assay, the measure of the peroxide rates was performed in different areas, three after UV radiation, two treated, one non-treated, and an other non-treated and non-irradiated. The peroxide presence was detected through a fluorescent probe on samples stripped form the sites above mentioned. The free-radical scavenger activity is calculated through the fluorimetry results. RESULTS: Based on non-irradiated areas data’s, the irradiated and complex-treated areas presented skin-peroxide concentration 116% lower, statistically significant (p=0,02% compared to the non-treated irradiated areas. Although, placebo-treated irradiated areas presented skin-peroxide concentration 49% lower, a non-statistically significant rate (p=0,501% compared to the irradiated areas. CONCLUSION: The results indicate that studied-complex, have significant skin-protective action against the free-radicals, usually formed after sun exposure.

  13. Chromatographic Methods for the Analysis of Polyphenols in Wines

    Directory of Open Access Journals (Sweden)

    Medić-Šarić, M.

    2009-03-01

    array detector (DAD. Enhancing selectivity and sensitivity for the determination of certain polyphenols requires the application of different detection techniques, such as fluorimetry (Fig. 4, electrochemistry, chemiluminescence, and/or mass spectrometry coupled with ionization techniques: electrospray (ESI, matrix-assisted laser desorption/ionization (MALDI, and atmospheric pressure chemical ionization (APCI.Gas chromatography (GC methods developed for the analysis of polyphenols require derivatization to the volatile compounds and mass-spectrometric detection in the selective ion-monitoring mode (GC/MS-SIM.A special technique, capillary electrophoresis (CE, is a powerful tool for the analysis of polyphenol contents of white and red wines, with an opportune sample preconcentration step. Micellar electrokinetic capillary chromatography (MECC has extended the utility of capillary electrophoresis to the separation of neutral analytes under the influence of an electric field. Fractionation of monomeric and polymeric pigments of higher molecular mass by gel permeation chromatography (GPC improved the analysis of these compounds by CE.

  14. Targeting the epidermal growth factor receptor in non-small cell lung cancer cells: the effect of combining RNA interference with tyrosine kinase inhibitors or cetuximab

    Directory of Open Access Journals (Sweden)

    Chen Gang

    2012-03-01

    Full Text Available Abstract Background The epidermal growth factor receptor (EGFR is a validated therapeutic target in non-small cell lung cancer (NSCLC. However, current single agent receptor targeting does not achieve a maximal therapeutic effect, and some mutations confer resistance to current available agents. In the current study we have examined, in different NSCLC cell lines, the combined effect of RNA interference targeting the EGFR mRNA, and inactivation of EGFR signaling using different receptor tyrosine kinase inhibitors (TKIs or a monoclonal antibody cetuximab. Methods NSCLC cells (cell lines HCC827, H292, H358, H1650, and H1975 were transfected with EGFR siRNA and/or treated with the TKIs gefitinib, erlotinib, and afatinib, and/or with the monoclonal antibody cetuximab. The reduction of EGFR mRNA expression was measured by real-time quantitative RT-PCR. The down-regulation of EGFR protein expression was measured by western blot, and the proliferation, viability, caspase3/7 activity, and apoptotic morphology were monitored by spectrophotometry, fluorimetry, and fluorescence microscopy. The combined effect of EGFR siRNA and different drugs was evaluated using a combination index. Results EGFR-specific siRNA strongly inhibited EGFR protein expression almost equally in all cell lines and inhibited cell growth and induced cell apoptosis in all NSCLC cell lines studied, albeit with a different magnitude. The effects on growth obtained with siRNA was strikingly different from the effects obtained with TKIs. The effects of siRNA probably correlate with the overall oncogenic significance of the receptor, which is only partly inhibited by the TKIs. The cells which showed weak response to TKIs, such as the H1975 cell line containing the T790M resistance mutation, were found to be responsive to siRNA knockdown of EGFR, as were cell lines with downstream TKI resistance mutations. The cell line HCC827, harboring an exon 19 deletion mutation, was more than 10-fold

  15. Effect of Schisandra chinensis polysaccharide on intracerebral acetylcholinesterase and monoamine neurotransmitters in a D-galactose-induced aging brain mouse model

    Institute of Scientific and Technical Information of China (English)

    Mingsan Miao; Jianlian Gao; Guangwei Zhang; Xiao Ma; Ying Zhang

    2009-01-01

    model group were intragastrically infused with the same volume of normal saline (0.2 mL/10 g) once per day for 30 consecutive days. MAIN OUTCOME MEASURES: Two hours after the final administration, pathohistological changes in the cerebral cortex and hippocampus were observed using hematoxylin & eosin staining. AChE activity was detected using chromatometry. Monoamine neurotransmitter content was measured using fluorimetry. Learning and memory was measured using the step down test and darkness avoidance test. RESULTS: Both Schisandra chinensis polysaccharide and Kangnaoling improved pathological injury to the cerebral cortex and hippocampus in a mouse model of brain aging. Compared with the blank control group, AChE activity and content of norepinephrine (NA), dopamine (DA), and 5-hydroxytryptamine (5-HT) were significantly decreased in the model group (P<0.01). In contrast, AChE activity and NA, DA, and 5-HT levels significantly increased in the Kangnaoling and high dosage Schisandra chinensis polysaccharide groups (P<0.01), while NA levels significantly increased in the low dosage Schisandra chinensis polysaccharide group (P<0.01). Drug treatment improved learning and memory abilities (P<0.01 or P<0.05). CONCLUSION: Schisandra chinensis polysaccharide significantly increased levels of central neurotransmitters and improved learning and memory in a mouse model of brain aging. The effects of Schisandra chinensis polysaccharide were equal to that of Kangnaoling pellets.

  16. Location of rare-earth dopants on LiCAF and LiSAF laser hosts via XRD, EXAFS and computer modeling technique

    International Nuclear Information System (INIS)

    Valerio, Mario Ernesto Giroldo; Amaral, Jomar Batista de; Baldochi, Sonia Licia Vera; Mazzocchi, L.; Sasaki, Jose Marcos; Jackson, Robert A.

    2004-01-01

    Full text: Cr-doped LiCaAlF 6 (LiCAF) and LiSrAlF 6 (LiSAF) were used as laser operating in the near infrared region. Ce-doped LiCAF and LiSAF have been reported as leading candidates for tunable all-solid-state lasers in the UV region. Spectroscopic properties of LiCaAlF 6 : Nd suggest that this crystal can be used as selective optical filter and refractive element for 157 nm photolithography. The question of whether the RE dopant will prefer the Li + , the M 2+ site or the Al 3+ site is not yet known. Nevertheless most of the optical properties of these hosts including their laser action depend on the local symmetry, charge compensation mechanism and possible deformation of the lattice. In the present work, Powder X-ray Diffraction (XRD), X-ray Absorption Spectroscopy (XAS), Spectro fluorimetry, combined with computer modeling, were used to study the local structure around the dopant and determine the site occupied by them and also the distance and nature of the co-ordinating atoms. The compounds were prepared from commercially available CaF2 and SrF2 powders of high purity; LiF previously purified by the zone melting method, and AlF3 and RE dopants obtained from the hydro fluorination of commercial Al 2 O 3 . The synthesis of 2 mol % RE doped LiCAF and LiSAF samples were performed in a platinum reactor. The compositions were 2 mol % LiF and AlF3 enriched to compensate for their high vaporization. Powder XRD measurements were performed at room temperature in a Rigaku DMAX diffractometer in step scan mode using Cu K radiation. The Rietveld method (DBWS-9807a software) was employed in the analysis of the patterns. It was found that in the doped samples the concentration of the LiCAF or LiSAF phases are 84-95% and a small amount of AlF 3 and α - Li 3 AlF 6 were formed. The XAS experiments were performed on and above the L III absorption edge of the Er, Ho and Nd ions in fluorescence and transmission mode at room temperature in the XAS station at the LNLS, Campinas

  17. Report on the intercomparison run for the determination of radionuclides in soils. IAEA-326 and IAEA-327

    International Nuclear Information System (INIS)

    Bojanowski, R.; Radecki, Z.; Campbell, M.J.; Burns, K.I.; Trinkl, A.

    2001-04-01

    to go beyond a purely statistical evaluation and give some thought to the quality of input data. As a result, the overall mean values for some radionuclides have changed markedly compared to the data obtained by the traditional approach followed in the Preliminary Evaluation Report IAEA/AL/100P. The main advantage of such a treatment lies in the considerable reduction of uncertainty range about the mean values. Whether or not these narrower confidence intervals will be challenged by new, more rigorous measurements remains to be seen. The participants can compare the more conservative estimates which are provided in the preliminary report, against the values reported here and judge for themselves the value of this approach. The Intercomparison revealed a number of methodological problems associated with radionuclide analysis. Some of them, like discrepant values for Ra-226 activity concentration due to improper sample preparation, interferences in alpha- and gamma- spectra and effects of incomplete sample decomposition, were already observed in earlier intercomparisons and have been addressed in the literature. This exercise has confirmed that these problems still exist and has also raised some interesting questions: Why do the analysts who employ HF for total sample dissolution fail to get results comparable with those obtained on fused samples? Why are results obtained by neutron activation analysis so scattered? Why do all results obtained by Fluorimetry fall beyond the range of accepted results? Why are there so many poor quality data for Pb-210 and Po-210 and for Sr-90 at the lower activity level (IAEA-327)? If the quality of environmental data for these radionuclides is to improve much remains to be done to improve the methodology of radiochemical operations and measurements. Unless reliable reference materials with narrow uncertainty ranges about their reference values are available, such problems have little chance to be solved and still others may remain

  18. Sorption of uranyl species on zircon and zirconia

    International Nuclear Information System (INIS)

    Lomenech, C.; Drot, R.; Simoni, E.; Ehrhardt, J.J.; Mielczarski, J.

    2002-01-01

    The safety of a long-term storage of radioactive waste in deep geological repositories would be strongly affected by the migration properties of radionuclides through the different barriers to the surface of the earth. Since the main process involved in the retention of radioactive ions is their sorption at the water/ mineral interface, a quantitative description of the sorption reactions is needed. Macroscopic data have for a long time been the only source of information used to propose a modelling of sorption equilibria, although they bring no direct information on the nature of the sorbed species; a microscopic structural investigation of the surface complexes is difficult indeed, because of the small amount of matter sorbed. Thus, in this study, parallel to the macroscopic measurements, different complementary spectroscopic techniques have been used in order to determine the nature of the surface species. As the final purpose of such a study is the simulation of the experimental retention data, the precise structural identification of the sorption equilibria will then be very useful to constrain the data simulation code. In this work, we present the results of both macroscopic and microscopic studies of the sorption of uranyl species on zircon and zirconia. The first part of our macroscopic approach was the surface characterisation of the non-sorbed materials by the determination of the specific areas, of the pH of the isoelectric points, and of the sorption site numbers, while the second part aimed at obtaining the sorption isotherms (percentage of sorption versus pH), which was performed using alpha spectrometry, for different uranyl concentrations, media (NaClO 4 or KNO 3 ) and ionic strengths. The spectroscopic identification of the different surface complexes and sorption sites has been carried out using four different spectroscopies. Whereas tune-resolved laser spectro-fluorimetry gave a direct answer concerning the number of surface species (only for a

  19. 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

    Crop-pest control is of major economic importance and demands the aid of the latest advances in science. Radioisotopes and radiation are being employed to increase the efficiency of existing insect pest control. They are extremely valuable, since improvements to existing methods depend on having detailed data on the bioecology, toxicology, and so on. Radioactive labelling of insects has been extremely promising in bioecology; the labelling of grain pests (Eurygaster integriceps Put., Hadena sordida Skh.) and grain-pest parasites (Meniscus agnatus Crow, Pseudogonia cinerascens Rond.) has provided information about their areas of migration, habitats, sizes of population and the feeding habits. The same technique was used to determine the rate of propagation of the Colorado beetle (Leptinotarsa decemlineota Say), which is subject to quarantine controls; subsequently an extermination programme was carried out on the basis of the data obtained. It also provides a valuable means of studying the extremely complex problems of parasitism and predaceousness, in particular intermediate feeding cycles and chemotaxis. The feeding areas of field rodents have been mapped out with the help of a self-labelling, radioactive-bait technique. Pesticides synthesized with radioisotopes have been used in conjunction with radiochromatography, fluorimetry and other techniques to study the highly complex biochemical processes caused to toxicants in plants and insects. It has also been possible to determine the rate of hydrolysis of organic-phosphorus insecticide compounds of the thiphos and metaphos type as a function of the degree of development and the physiological state of plants as well as of environmental conditions. Data have been obtained on the length of time residual quantities of toxicants are retained in agriculture products following different periods of chemical treatment. Radioisotope techniques have yielded information on various metabolic processes exhibiting different