Sample records for eosin-5-maleimide flow cytometric
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Arora, R D; Dass, J; Maydeo, S; Arya, V; Radhakrishnan, N; Sachdeva, A; Kotwal, J; Bhargava, M
2018-03-24
Hereditary spherocytosis (HS) is the most common inherited hemolytic anemia with heterogeneous clinico-laboratory manifestations. We evaluated the flow-cytometric tests: eosin-5'-maleimide (EMA) and flow-cytometric osmotic fragility test (FOFT) and the conventional osmotic fragility tests (OFT) for the diagnosis of hereditary spherocytosis (HS). One hundred two suspected HS patients underwent EMA, FOFT, incubated OFT (IOFT), and room temperature OFT (RT-OFT). In addition, 10 cases of immune hemolytic anemia (IHA) were included, and performance of the above 4 tests was evaluated. For EMA and FOFT, 5 normal controls were assessed together with the patients and cutoffs were calculated using receiver-operator-characteristics curve (ROC) analysis. The best cutoff for %EMA decrease was 12.5%, and for FOFT, %residual red cells (%RRC) was 25.6%. The sensitivity and specificity of RT-OFT was 62.06% and 86.3%, respectively, while that of IOFT was 79.31% and 87.67%, respectively. Both flow cytometric tests performed better. Sensitivity and specificity of EMA was 86.2% and 93.9% respectively, and that of FOFT was 96.6% and 98.63%, respectively. The combination of the FOFT with IOFT or EMA dye-binding test yields a sensitivity of 100%, but with EMA, it had a higher specificity. Hb/MCHC was a predictor of the severity of the disease while %EMA decrease and %RRC did not correlate with severity of the disease. Flow-cytometric osmotic fragility test is the best possible single test followed by EMA for diagnosis of HS. A combination of FOFT and EMA can correctly diagnose 100% patients. These tests are likely to replace conventional OFTs in future. © 2018 John Wiley & Sons Ltd.
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Mehra, Simmi; Tyagi, Neetu; Dorwal, Pranav; Pande, Amit; Jain, Dharmendra; Sachdev, Ritesh; Raina, Vimarsh
2015-06-01
The eosin-5'-maleimide (EMA) binding test using flow cytometry is a common method to measure reduced mean channel fluorescence (MCF) of EMA-labeled red blood cells (RBCs) from patients with red cell membrane disorders. The basic principle of the EMA-RBC binding test involves the covalent binding of EMA to lysine-430 on the first extracellular loop of band 3 protein. In the present study, the MCF of EMA was analyzed for samples derived from 12 healthy volunteers (controls) to determine the stability (i.e., the percentage decrease in fluorescence) of EMA over a period of 1 year. Comparison of periodical MCF readings over time, that is, at 2-month intervals, showed that there were no significant changes in mean channel fluorescence for up to 6 months; however, there was a significant decrease in MCF at 8 months. For optimal dye utilization, EMA remained stable only for up to 6 months. Therefore, we recommend reconstitution of the dye every 6 months when implementing this test and storage at -80℃ in dark conditions.
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Wang, Jiying; Zheng, Bin; Zhao, Yuping; Chen, Xuejing; Liu, Yan; Bo, Lijin; Zheng, Yizhou; Zhang, Fengkui; Ru, Kun; Wang, Huijun
2015-07-01
To investigate the sensitivity and specificity of eosin-5'-maleimide (EMA)assay for the diagnosis of hereditary spherocytosis (HS), and to verify the stability of reagent and samples. EMA flow cytometry test, NaCl-osmotic fragility test and acidified glycerol lysis test were performed using peripheral blood samples from 80 patients with HS and 44 patients with other blood diseases, the sensitivity and specificity of the three methods were compared, and the feasibility of EMA binding test was estimated. The stability of EMA reagent and HS samples stored at different temperatures were tested. Among the 124 tested samples, the sensitivity and specificity of EMA binding test was 0.925 and 0.954, that of NaCl-osmotic fragility test was 0.950 and 0.455, and that of acidified glycerol lysis test was 1.000 and 0.318, respectively. Although the sensitivity of NaCl-osmotic fragility test and acidified glycerol lysis test was a little higher than that of EMA binding test, the specificity of the former two methods was poor, they couldn't clearly distinguish whether spherocytosis is hereditary spherocytosis. The experiment results showed that EMA was sensitive to the temperature and should not be stored in a small aliquots at -80 ℃ over a period of 6 months. The stability of the HS sample was better, 6 days storage at 4 ℃ and 3 days storage at room temperature had no influence on the results. EMA binding test by flow cytometry showed good sensitivity and specificity for HS diagnosis. EMA reagent should be stored at-80 ℃ and the HS samples should be tested within 6 days storage at 4 ℃ and 3 days at room temperature.
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Analysis of Hereditary Elliptocytosis with Decreased Binding of Eosin-5-maleimide to Red Blood Cells
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Shin-ichiro Suemori
2015-01-01
Full Text Available Flow cytometric test for analyzing the eosin-5-maleimide (EMA binding to red blood cells has been believed to be a specific method for diagnosing hereditary spherocytosis (HS. However, it has been reported that diseases other than HS, such as hereditary pyropoikilocytosis (HPP and Southeast Asian ovalocytosis (SAO, which are forms in the category of hereditary elliptocytosis (HE, show decreased EMA binding to red blood cells. We analyzed EMA binding to red blood cells in 101 healthy control subjects and 42 HS patients and obtained a mean channel fluorescence (MCF cut-off value of 36.4 (sensitivity 0.97, specificity 0.95. Using this method, we also analyzed 12 HE patients. Among them, four HE patients showed the MCF at or below the cut-off value. It indicates that some HE patients have decreased EMA binding to red blood cells. Two of these four HE patients were classified as common HE, and two were spherocytic HE with reduced spectrin. This study demonstrates that, in addition to patients with HPP or SAO, some HE patients have decreased EMA binding to red blood cells.
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[THE CYTOMETRIC TECHNIQUE OF BINDING OF EOSIN-5-MALEIMIDE IN DIAGNOSTIC OF INHERENT SPHEROCYTOSIS].
Kuzminova, J A; Plyasunova, S A; Jogov, V V; Smetanina, N S
2016-03-01
The laboratory diagnostic of inherent spherocytosis is based on detection of spherocytes in peripheral blood, decreasing of index of sphericity, decreasing of osmotic resistance of erythrocytes. The new test of diagnostic of hereditary spherocytosis build on molecular defect was developed on the basis of binding extracellular fragments of protein of band 3 with eosin-5-maleimide (EMA-test). The study was carried out to implement comparative analysis of sensitivity and specificity of techniques applied to diagnose inherent spherocytosis. The sampling of 94 patients with various forms of anemias was analyzed All patients were applied complex clinical laboratory examination including analysis of osmotic resistance of erythrocytes, erythrocytometry and EMA-test as specific techniques of diagnostic of inherent spherocytosis. In 51 out of 94 patients (54%) decreasing of values of EMA-test was detected and in 47 patients diagnosis of inherent spherocytosis was confirmed. The standard values of EMA-test were established in 43 patients (46%) and 12 patients out of them with established diagnosis of inherent spherocytosis. Therefore, sensitivity of EMA-test made up to 79% and specificity - 80%. The most sensitive techniques of diagnostic remain osmotic resistance of erythrocytes (91%) and index of sphericity (up to 96%). But the highest specificity in this respect has EMA-test (80%). Nowadays, none of implemented techniques of diagnostic of inherent spherocytosis can be applied as a universal one. The implementation of complex examination is needed for proper diagnostic of disease.
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Christensen, R D; Agarwal, A M; Nussenzveig, R H; Heikal, N; Liew, M A; Yaish, H M
2015-05-01
Neonates with undiagnosed hereditary spherocytosis (HS) are at risk for developing hazardous hyperbilirubinemia and anemia. Making an early diagnosis of HS in a neonate can prompt anticipatory guidance to prevent these adverse outcomes. A recent comparison study showed that a relatively new diagnostic test for HS, eosin-5-maleimide (EMA)-flow cytometry, performs better than other available tests in confirming HS. However, reports have not specifically examined the performance of this test among neonates. We compared EMA-flow cytometry from blood samples of healthy control neonates vs samples from neonates suspected of having HS on the basis of severe Coombs-negative jaundice and spherocytes on blood film. The diagnosis of HS was later either confirmed or excluded based on clinical findings and next generation sequencing (NGS) after which we correlated the EMA-flow results with the diagnosis. EMA-flow was performed on the blood of 31 neonates; 20 healthy term newborns and 11 who were suspected of having HS. Eight of the 11 were later confirmed positive for HS and one was confirmed positive for hereditary elliptocytosis (HE). All nine had persistently abnormal erythroid morphology, reticulocytosis and anemia, and eight of the nine had relevant mutations discovered using NGS. The other was confirmed positive for HS on the basis that a parent had HS, and the neonate's spherocytosis, reticulocytosis and anemia persisted. The 20 healthy controls and the 2 in whom HS was initially suspected but later excluded all had EMA-flow results in the range reported in healthy children and adults. In contrast, all nine in whom HS or HE was confirmed had abnormal EMA-flow results consistent with previous reports in older children and adults with HS. Although our sample size is small, our findings are consistent with the literature in older children and adults suggesting that EMA-flow cytometric testing performs well in supporting the diagnosis of HS/HE during the early neonatal
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Joshi, P; Aggarwal, A; Jamwal, M; Sachdeva, M U S; Bansal, D; Malhotra, P; Sharma, P; Das, R
2016-10-01
Laboratory diagnosis of hereditary spherocytosis (HS) relies on increased incubated red cell osmotic fragility test for screening. We evaluated the diagnostic role of eosin-5'-maleimide (EMA) binding test by flow cytometry in spherocytic and microcytic hypochromic hematological disorders in North Indians. EMA binding test using flow cytometry was performed on 55 HS (40 families), 26 iron deficiency anemia (IDA), 32 β-thalassemia trait (βTT), and 10 autoimmune hemolytic anemia (AIHA) cases and 121 normals. Mean channel fluorescence (MCF) and coefficient of variation (CV) were studied. Different MCF parameters (MCF, MCF ratio, percent decrease MCF) and percent increase in CV were analyzed. Receiver operating characteristics analysis was performed to determine best cutoff values, sensitivity, and specificity for discriminating HS from other red cell disorders. MCF ratio of HS group was significantly lower than normals (0.67 ± 0.07 vs. 1.01 ± 0.05, P < 0.001) and other cases. All patients with HS showed MCF ratio to be ≤0.79. Four postsplenectomy cases with near-normal hemograms also revealed low MCF ratio, showing the specificity of the test. EMA assay was efficient to diagnose cases of HS including postsplenectomy cases and shows no overlap with IDA, βTT, and AIHA. © 2016 John Wiley & Sons Ltd.
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Ciepiela, Olga; Adamowicz-Salach, Anna; Bystrzycka, Weronika; Łukasik, Jan; Kotuła, Iwona
2015-08-01
Eosin-5'-maleimide (EMA) binding test is a flow cytometric test used to detect hereditary spherocytosis (HS). To perform the test sample from patients, 5-6 reference samples of red blood are needed. Our aim was to investigate how the mean corpuscular volume (MCV) of red blood cells influences on the value of fluorescence of bounded EMA dye and how the choice of reference samples affects the test result. EMA test was performed in peripheral blood from 404 individuals, including 31 children suffering from HS. Mean fluorescence channel of EMA-RBCs was measured with Cytomics FC500 flow cytometer. Mean corpuscular volume of RBCs was assessed with LH750 Beckman Coulter. Statistical analysis was performed using Graph Pad Prism. The correlation Spearman coefficient between mean channel of fluorescence of EMA-RBCs and MCV was r = 0.39, p < 0.0001. Interpretation of EMA test depends on MCV of the reference samples. If reference blood samples have lower MCV than the patients MCV, EMA test result might be negative. Due to different MCV values of RBCs in infancy and ca. Three months later, EMA test in neonates might be interpreted falsely negative. Samples from children younger than 3 months old had EMA test result 86.1 ± 11.7 %, whereas same samples that analyzed 4.1 ± 2.1 later had results of 75.4 ± 4.5 %, p < 0.05. Mean fluorescence of EMA-bound RBC depends on RBC's volume. MCV of reference samples affects EMA test results; thus, we recommend selection of reference samples with MCV in range of ±2 fL compared to MCV of patient RBC's.
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The Gas-Phase Photophysics of Eosin Y and its Maleimide Conjugate.
Daly, Steven; Kulesza, Alexander; Knight, Geoffrey; MacAleese, Luke; Antoine, Rodolphe; Dugourd, Philippe
2016-05-26
The use of the xanthene family of dyes as fluorescent probes in a wide range of applications has provided impetus for the studying of their photophysical properties. In particular, recent advances in gas-phase techniques such as FRET that utilize such chromophores have placed a greater importance on the characterization of these properties in the gas phase. Additionally, the use of synthetic linker chains to graft the chromophores in a site-specific manner to their target system is ubiquitous. There is, however, often limited information on how the addition of such a linker chain may affect the photophysical properties of the chromophores, which is of fundamental importance for interpretation of experimental data reliant on grafted chromophores. Here, we present data on the optical spectroscopy of different protonation states of Eosin Y, a fluorescein derivative. We compare the photophysics of Eosin Y to its maleimide conjugate, and to the thioether product of the reaction of this conjugate with cysteamine. Comparison of the mass spectra following laser irradiation shows that very different relaxation takes place upon addition of the maleimide moiety but that the photophysics of the bare chromophore are restored upon addition of cysteamine. This radical change in the photophysics is interpreted in terms of charge-transfer states, whose energy relative to the S1 ← S0 transition of the chromophore is dependent on the conjugation of the maleimide moiety. We also show that the shape of the absorption band is unchanged in the gas-phase as compared to the solution-phase, showing a maximum with a shoulder toward the blue, and examination of isotope distributions of the isolated ions show that this shoulder cannot be due to the presence of dimers. Consideration of the fluorescence emission spectrum allows a tentative assignment of the shoulder to be due to a vibrational progression with a high Franck-Condon factor.
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Blackman, S M; Cobb, C E; Beth, A H; Piston, D W
1996-01-01
The dominant motional mode for membrane proteins is uniaxial rotational diffusion about the membrane normal axis, and investigations of their rotational dynamics can yield insight into both the oligomeric state of the protein and its interactions with other proteins such as the cytoskeleton. However, results from the spectroscopic methods used to study these dynamics are dependent on the orientation of the probe relative to the axis of motion. We have employed polarized fluorescence confocal microscopy to measure the orientation of eosin-5-maleimide covalently reacted with Lys-430 of human erythrocyte band 3. Steady-state polarized fluorescence images showed distinct intensity patterns, which were fit to an orientation distribution of the eosin absorption and emission dipoles relative to the membrane normal axis. This orientation was found to be unchanged by trypsin treatment, which cleaves band 3 between the integral membrane domain and the cytoskeleton-attached domain. this result suggests that phosphorescence anisotropy changes observed after trypsin treatment are due to a rotational constraint change rather than a reorientation of eosin. By coupling time-resolved prompt fluorescence anisotropy with confocal microscopy, we calculated the expected amplitudes of the e-Dt and e-4Dt terms from the uniaxial rotational diffusion model and found that the e-4Dt term should dominate the anisotropy decay. Delayed fluorescence and phosphorescence anisotropy decays of control and trypsin-treated band 3 in ghosts, analyzed as multiple uniaxially rotating populations using the amplitudes predicted by confocal microscopy, were consistent with three motional species with uniaxial correlation times ranging from 7 microseconds to 1.4 ms. Images FIGURE 4 FIGURE 8 FIGURE 9 PMID:8804603
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Agarwal, Archana M; Liew, Michael A; Nussenzveig, Roberto H; Sangle, Nikhil; Heikal, Nahla; Yaish, Hassan; Christensen, Robert
2016-11-01
The eosin-5'maleimide (EMA) binding test has been studied extensively for the detection of hereditary spherocytosis (HS). Its performance characteristics have been compared to NaCl-based or glycerol lysis-based red cell osmotic fragility tests and cryohemolysis. HS samples are also better identified when both mean channel fluorescence (MCF) of EMA relative to controls and the coefficient of variation (CV) are analyzed. We looked at 65 normal controls including 30 adults 25-65 years old and 35 newborns and 12 HS cases. In addition to the MCF and the CV, we used a side scatter (SSC) vs. EMA fluorescence gate or "footprint" to depict where normal erythrocytes should appear. Erythrocytes that have reduced band 3 protein appear outside of the footprint. In our study, newborn data did not cluster with the samples from working age individuals. The MCF and the CVs of normal newborns were higher than normal adult group. However, the footprint data of normal samples relative to their controls was around 99.5% for each group, because the footprint was moved to fit the pattern of the normal. The inclusion of footprint parameter will help in better standardization as well as implementation of this test across different age groups as well as different instruments. © 2015 International Clinical Cytometry Society. © 2015 International Clinical Cytometry Society.
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Liang, Zhongwei; Xu, Song; Tian, Wenyan; Zhang, Ronghua
2015-01-01
A novel and simple strategy for the efficient synthesis of the corresponding tetrahydroquinolines from N,N-dimethylanilines and maleimides using visible light in an air atmosphere in the presence of Eosin Y as a photocatalyst has been developed. The metal-free protocol involves aerobic oxidative cyclization via sp(3) C-H bond functionalization process to afford good yields in a one-pot procedure under mild conditions.
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Directory of Open Access Journals (Sweden)
Zhongwei Liang
2015-04-01
Full Text Available A novel and simple strategy for the efficient synthesis of the corresponding tetrahydroquinolines from N,N-dimethylanilines and maleimides using visible light in an air atmosphere in the presence of Eosin Y as a photocatalyst has been developed. The metal-free protocol involves aerobic oxidative cyclization via sp3 C–H bond functionalization process to afford good yields in a one-pot procedure under mild conditions.
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Ciepiela, Olga; Adamowicz-Salach, Anna; Zdziechowicz, Izabela; Kotuła, Iwona
2016-11-01
Diagnosis of hereditary spherocytosis (HS) is based on clinical evaluation and eosin-5'-maleimide (EMA) test. A decrease in EMA fluorescence compared with healthy individuals is typical for HS and serves as a basis for HS diagnosis. Sensitivity and specificity of the test is high and false-positive results rarely occur. Studies have shown that anticoagulated blood sample when stored at 4°C for 7 days do not affect the test results. This case study is about an autoimmune hemolytic anemia patient who showed a primary positive result for EMA test (decrease in EMA fluorescence-47% compared with 100% for samples of healthy individual), when the test was performed in the sample stored for 48 hours after venipuncture and before staining. An irrelevant decrease (92.5% compared with 100% for samples of healthy individual) was found when freshly collected sample was analyzed. On the basis of the results obtained, it is recommended that EMA staining should be performed on the same day of blood collection for patients with significant hemolysis.
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Andres, Oliver; Eber, Stefan; Speer, Christian P
2015-12-01
Exact diagnosis of hereditary spherocytosis (HS) is widely considered unreliable around birth. However, early postnatal diagnosis at the beginning of congenital hemolysis may be essential for managing neonatal anemia and hemolytic icterus, identifying those at high risk for severe hyperbilirubinemia, irreversible kernicterus, or sudden need for red cell transfusion. We analyzed 37 blood samples from neonates or infants up to six weeks of life that had been collected in-house or shipped to our laboratory due to suspected red cell membrane disorder. By combining assessment of red cell morphology, acidified glycerol lysis test (AGLT), and eosin-5'-maleimide (EMA) binding assay, we were able to clearly exclude HS in 22 and confirm HS in 10 patients, of which one had undergone red cell transfusion prior to blood sampling. Assessment of red cell morphology and normal test results allowed diagnosis of infantile pyknocytosis or Heinz body anemia in three neonates. Re-evaluation of five patients with inconsistent results of AGLT and EMA binding led to confirmation of HS in two cases. Automated analysis of hematologic parameters revealed elevated proportion of hyperdense cells to be a highly significant indicator for HS in neonatal infants. We showed that assessment of red cell morphology in combination with AGLT and EMA binding assay is a reliable basis for confirming or rejecting suspected diagnosis of HS even in neonates. Our data underline the necessity for blood sampling and laboratory exploration in suspected red cell membrane or enzyme defects at the earliest occasion.
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Hunt, Linda; Greenwood, David; Heimpel, Hermann; Noel, Nigel; Whiteway, Alastair; King, May-Jean
2015-01-01
The eosin-5'-maleimide (EMA) Binding test measures reduced mean channel fluorescence (MCF) reading of EMA-labeled red cells (EMA-RBCs) from patients with hereditary spherocytosis (HS). Reporting test results can be either in the actual MCF reading or as a ratio by normalization of the test MCF result to the mean MCF value of six normal controls. The latter format has potential for universal reporting. We analyzed three years' archival MCF data from HS and non-HS patient groups for establishment of reference ranges of ratios for normal adults and HS. A prospective study used FC500 and FACS Canto II cytometers to analyze contemporaneously EMA-RBCs from several patient groups and normal donors. Statistical analyses of the prospective data determined the cut-off values, and the sensitivity and specificity for HS respectively for the MCF and the ratio result presentations. The effect of using fewer than six normal controls for the ratio denominator was explored. The FC500 gave a mean ratio of 0.782 (SD=0.086) in HS patients with an optimal cut-off ratio of 0.918 (98.7% specificity, 95.6% sensitivity), and gray area ratio of 0.868-0.918. The Canto II gave a mean ratio of 0.774 (SD=0.085) with an optimal cut-off ratio of 0.925 (97.1% specificity and 100% sensitivity), and gray area ratio of 0.859-0.925. Harmonization of result presentation is feasible with no apparent constraint by instrument design. Interpretation of gray-area data requires an assessment of patient's clinical presentation and family history or performing a family study. © 2014 Clinical Cytometry Society.
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Peng, G X; Yang, W R; Jing, L P; Zhang, L; Zhou, K; Li, Y; Ye, L; Li, Y; Li, J P; Fan, H H; Song, L; Zhao, X; Wu, Z J; Yang, Y; Xiong, Y Z; Wang, H J; Zhang, F K
2017-06-14
Objective: To investigate the relationship between the eosin-5'-maleimide (EMA) binding test and the clinical severity of hereditary spherocytosis (HS). Methods: A total of 258 un-splenectomize HS patients were consecutively enrolled. Correlation of hemoglobin concentration, hemolytic parameters, compensating erythropoiesis and the EMA binding test were evaluated. Results: 258 (128 male and 130 female) patients were included in this study, including 91 compensatory hemolysis patients, 53 patients with mild anemia, 78 patients with moderate anemia and 36 patients with severe anemia. The median age at diagnosis was 23 (2-70) years. The median decreased fluorescence intensity of EMA binding test was 29.97% (16.09%-47.34%) and the average intensity was (29.70±6.28) % of 258 HS patients. The decreased EMA binding fluorescence intensity correlated with MCV ( r =-0.343, P <0.001) and MCHC ( r =0.223, P <0.001). There was no relationship between EMA fluorescence intensity and absolute reticulocyte count ( r =0.080, P =0.198) , reticulocyte percentile ( r =-0.015, P =0.813) , IBIL levels ( r =-0.009, P =0.902) , HGB levels ( r =-0.067, P =0.280). Evaluated as a quartile variable, EMA fluorescence intensity was not correlated with anemia severity ( C =0.150, P =0.746). Conclusion: EMA binding test does not related to anemia levels and has no major clinical implications for disease severity in HS.
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Quantifying Distribution of Flow Cytometric TCR-Vβ Usage with Economic Statistics.
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Kornelis S M van der Geest
Full Text Available Measuring changes of the T cell receptor (TCR repertoire is important to many fields of medicine. Flow cytometry is a popular technique to study the TCR repertoire, as it quickly provides insight into the TCR-Vβ usage among well-defined populations of T cells. However, the interpretation of the flow cytometric data remains difficult, and subtle TCR repertoire changes may go undetected. Here, we introduce a novel means for analyzing the flow cytometric data on TCR-Vβ usage. By applying economic statistics, we calculated the Gini-TCR skewing index from the flow cytometric TCR-Vβ analysis. The Gini-TCR skewing index, which is a direct measure of TCR-Vβ distribution among T cells, allowed us to track subtle changes of the TCR repertoire among distinct populations of T cells. Application of the Gini-TCR skewing index to the flow cytometric TCR-Vβ analysis will greatly help to gain better understanding of the TCR repertoire in health and disease.
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Buysschaert, Benjamin; Kerckhof, Frederiek-Maarten; Vandamme, Peter; De Baets, Bernard; Boon, Nico
2018-02-01
The analysis of microbial populations is fundamental, not only for developing a deeper understanding of microbial communities but also for their engineering in biotechnological applications. Many methods have been developed to study their characteristics and over the last few decades, molecular analysis tools, such as DNA sequencing, have been used with considerable success to identify the composition of microbial populations. Recently, flow cytometric fingerprinting is emerging as a promising and powerful method to analyze bacterial populations. So far, these methods have primarily been used to observe shifts in the composition of microbial communities of natural samples. In this article, we apply a flow cytometric fingerprinting method to discriminate among 29 Lactobacillus strains. Our results indicate that it is possible to discriminate among 27 Lactobacillus strains by staining with SYBR green I and that the discriminatory power can be increased by combined SYBR green I and propidium iodide staining. Furthermore, we illustrate the impact of physiological changes on the fingerprinting method by demonstrating how flow cytometric fingerprinting is able to discriminate the different growth phases of a microbial culture. The sensitivity of the method is assessed by its ability to detect changes in the relative abundance of a mix of polystyrene beads down to 1.2%. When a mix of bacteria was used, the sensitivity was as between 1.2% and 5%. The presented data demonstrate that flow cytometric fingerprinting is a sensitive and reproducible technique with the potential to be applied as a method for the dereplication of bacterial isolates. © 2017 International Society for Advancement of Cytometry. © 2017 International Society for Advancement of Cytometry.
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Uncovering Aberrant Mutant PKA Function with Flow Cytometric FRET
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Shin-Rong Lee
2016-03-01
Full Text Available Biology has been revolutionized by tools that allow the detection and characterization of protein-protein interactions (PPIs. Förster resonance energy transfer (FRET-based methods have become particularly attractive as they allow quantitative studies of PPIs within the convenient and relevant context of living cells. We describe here an approach that allows the rapid construction of live-cell FRET-based binding curves using a commercially available flow cytometer. We illustrate a simple method for absolutely calibrating the cytometer, validating our binding assay against the gold standard isothermal calorimetry (ITC, and using flow cytometric FRET to uncover the structural and functional effects of the Cushing-syndrome-causing mutation (L206R on PKA’s catalytic subunit. We discover that this mutation not only differentially affects PKAcat’s binding to its multiple partners but also impacts its rate of catalysis. These findings improve our mechanistic understanding of this disease-causing mutation, while illustrating the simplicity, general applicability, and power of flow cytometric FRET.
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A flow cytometric assay for simultaneously measuring the ...
African Journals Online (AJOL)
This research objective was to exploit a novel method for measuring the proliferation, cytotoxicity of cytokine-induced killer (CIK) cells using carboxyfluorescein succinimidyl ester/proliferation index (CFSE/PI) and flow cytometric assay. As cells divide, CFSE is apportioned equally between the two daughter cells, leading to a ...
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International Nuclear Information System (INIS)
Yu, Hye-Weon; Kim, In S.; Niessner, Reinhard; Knopp, Dietmar
2012-01-01
Highlights: ► First time, duplex competitive bead-based flow cytometric immunoassay was developed using ODs. ► Antibody-coated QD detection probes and antigen-immobilized microspheres were synthesized. ► The two model target analytes were low molecular weight compounds of microbial and chemical origin. ► The determination of different water types was possible after simple filtration of samples. - Abstract: In answer to the ever-increasing need to perform the simultaneous analysis of environmental hazards, microcarrier-based multiplex technologies show great promise. Further integration with biofunctionalized quantum dots (QDs) creates new opportunities to extend the capabilities of multicolor flow cytometry with their unique fluorescence properties. Here, we have developed a competitive microbead-based flow cytometric immunoassay using QDs fluorescent labels for simultaneous detection of two analytes, bringing the benefits of sensitive, rapid and easy-of-manipulation analytical tool for environmental contaminants. As model target compounds, the cyanobacterial toxin microcystin-LR and the polycyclic aromatic hydrocarbon compound benzo[a]pyrene were selected. The assay was carried out in two steps: the competitive immunological reaction of multiple targets using their exclusive sensing elements of QD/antibody detection probes and antigen-coated microsphere, and the subsequent flow cytometric analysis. The fluorescence of the QD-encoded microsphere was thus found to be inversely proportional to target analyte concentration. Under optimized conditions, the proposed assay performed well within 30 min for the identification and quantitative analysis of the two environmental contaminants. For microcystin-LR and benzo[a]pyrene, dose–response curves with IC 50 values of 5 μg L −1 and 1.1 μg L −1 and dynamic ranges of 0.52–30 μg L −1 and 0.13–10 μg L −1 were obtained, respectively. Recovery was 92.6–106.5% for 5 types of water samples like bottled
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A flow cytometric assay for simultaneously measuring the ...
African Journals Online (AJOL)
Jane
2011-10-24
Oct 24, 2011 ... daughter cells, leading to a characteristic flow cytometric profile where a ... cell recognition without any impact on bone marrow hemato- ... cells of various cancer cells that load CFSE concentration ... (B) Target cells (R1) were further analyzed in an FL1/FL3 dot plot, ..... hematopoietic cell transplantation.
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International Nuclear Information System (INIS)
Kingsmore, S.F.; Crockard, A.D.; Fay, A.C.; McNeill, T.A.; Roberts, S.D.; Thompson, J.M.
1988-01-01
Several flow cytometric methods for the measurement of circulating immune complexes (CIC) have recently become available. We report a Raji cell flow cytometric assay (FCMA) that uses aggregated human globulin (AHG) as primary calibrator. Technical advantages of the Raji cell flow cytometric assay are discussed, and its clinical usefulness is evaluated in a method comparison study with the widely used Raji cell immunoradiometric assay. FCMA is more precise and has greater analytic sensitivity for AHG. Diagnostic sensitivity by the flow cytometric method is superior in systemic lupus erythematosus (SLE), rheumatoid arthritis, and vasculitis patients: however, diagnostic specificity is similar for both assays, but the reference interval of FCMA is narrower. Significant correlations were found between CIC levels obtained with both methods in SLE, rheumatoid arthritis, and vasculitis patients and in longitudinal studies of two patients with cerebral SLE. The Raji cell FCMA is recommended for measurement of CIC levels to clinical laboratories with access to a flow cytometer
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Directory of Open Access Journals (Sweden)
Li T
2013-10-01
Full Text Available Tianshu Li, Shinji TakeokaDepartment of Life Science and Medical Bioscience, Graduate School of Advanced Science and Engineering, Waseda University (TWIns, Shinjuku-ku, Tokyo, JapanAbstract: Maleimide is a stable and easy-to-handle moiety that rapidly and covalently conjugates thiol groups of cysteine residues in proteins or peptides. Herein, we use maleimide to modify the surface of liposomes in order to obtain an advanced drug delivery system. Employing a small amount (0.3 mol% of maleimide-polyethylene glycol (PEG to modify the surface of the liposomes M-GGLG-liposomes, composed of 1,5-dihexadecyl N,N-diglutamyl-lysyl-L-glutamate (GGLG/cholesterol/poly(ethylene glycol 1,2-distearoyl-sn-glycero-3-phosphoethanolamine (PEG5000-DSPE/maleimide-PEG5000-Glu2C18 at a molar ratio of 5:5:0.03:0.03, drug delivery efficiency was remarkably improved both in vitro and in vivo compared to unmodified liposomes (GGLG-liposomes, composed of GGLG/cholesterol/PEG5000-DSPE/PEG5000-Glu2C18 at a molar ratio of 5:5:0.03:0.03. Moreover, this modification did not elicit any detectable increase in cytotoxicity. The maleimide-modification did not alter the physical characteristics of the liposomes such as size, zeta potential, pH sensitivity, dispersibility and drug encapsulation efficiency. However, M-GGLG-liposomes were more rapidly (≥2-fold internalized into HeLa, HCC1954, and MDA-MB-468 cells compared to GGLG-liposomes. In vivo, M-GGLG-liposomes encapsulating doxorubicin (M-GGLG-DOX-liposomes also showed a more potent antitumor effect than GGLG-DOX-liposomes and the widely used 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC-DOX-liposomes after two subcutaneous injections around breast cancer tissue in mice. The biodistribution of liposomes in this model was observed using an in vivo imaging system, which showed that M-GGLG-liposomes were present for significantly longer at the injection site compared to GGLG-liposomes. The outstanding biological functions of
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Joetzke, Alexa E; Eberle, Nina; Nolte, Ingo; Mischke, Reinhard; Simon, Daniela
2012-06-01
To determine whether the extent of disease in dogs with lymphoma can be assessed via flow cytometry and to evaluate the suitability of fine-needle aspirates from the liver and spleen of dogs for flow cytometric examination. 44 dogs with multicentric B-cell (n = 35) or T-cell lymphoma (9) and 5 healthy control dogs. Procedures-Peripheral blood and bone marrow samples and fine-needle aspirates of lymph node, liver, and spleen were examined via flow cytometry. Logarithmically transformed T-cell-to-B-cell percentage ratio (log[T:B]) values were calculated. Thresholds defined by use of log(T:B) values of samples from control dogs were used to determine extranodal lymphoma involvement in lymphoma-affected dogs; results were compared with cytologic findings. 12 of 245 (5%) samples (9 liver, 1 spleen, and 2 bone marrow) had insufficient cellularity for flow cytometric evaluation. Mean log(T:B) values of samples from dogs with B-cell lymphoma were significantly lower than those of samples from the same site in dogs with T-cell lymphoma and in control dogs. In dogs with T-cell lymphoma, the log(T:B) of lymph node, bone marrow, and spleen samples was significantly higher than in control dogs. Of 165 samples assessed for extranodal lymphoma involvement, 116 (70%) tested positive via flow cytometric analysis; results agreed with cytologic findings in 133 of 161 (83%) samples evaluated via both methods. Results suggested that flow cytometry may aid in detection of extranodal lymphoma involvement in dogs, but further research is needed. Most fine-needle aspirates of liver and spleen were suitable for flow cytometric evaluation.
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Leukocytospermia and sperm preparation - a flow cytometric study
Directory of Open Access Journals (Sweden)
Perticarari Sandra
2009-01-01
Full Text Available Abstract Background Leukocytes represent the predominant source of reactive oxygen species both in seminal plasma and in sperm suspensions and have been demonstrated to negatively influence sperm function and fertilization rate in assisted reproduction procedures. Peroxidase test is the standard method recommended by WHO to detect semen leukocytes but it may be inaccurate. The aims of this study were (i to compare the efficiency of swim-up and density-gradient centrifugation techniques in removing seminal leukocytes, (ii to examine the effect of leukocytes on sperm preparation, and (iii to compare flow cytometry and peroxidase test in determining leukocyte concentration in semen using a multiparameter flow cytometric method. Methods Semen samples from 126 male partners of couples undergoing infertility investigations were analyzed for leukocytospermia using standard optical microscopy and flow cytometry. Sixty-nine out of 126 samples were also processed using simultaneously the swim-up and density-gradient centrifugation techniques. A multiparameter flow cytometric analysis to assess simultaneously sperm concentration, sperm viability, sperm apoptosis, and leukocyte concentration was carried out on neat and prepared sperm. Results Both sperm preparation methods removed most seminal leukocytes. However, the concentration of leukocytes was significantly lower after swim-up compared to that after density-gradient centrifugation preparation. Leukocytes concentration, either initial or in prepared fractions, was not correlated with sperm parameters (optical microscopy and flow cytometry parameters after semen processing. There was no correlation between leukocyte concentration in the ejaculate and sperm recovery rate, whereas a significant correlation was found between the concentration of the residual leukocytes in prepared fractions and viable sperm recovery rate. Although the overall concordance between the flow cytometry and the optical
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A flow cytometric assay technology based on quantum dots-encoded beads
International Nuclear Information System (INIS)
Wang Haiqiao; Liu Tiancai; Cao Yuancheng; Huang Zhenli; Wang Jianhao; Li Xiuqing; Zhao Yuandi
2006-01-01
A flow cytometric detecting technology based on quantum dots (QDs)-encoded beads has been described. Using this technology, several QDs-encoded beads with different code were identified effectively, and the target molecule (DNA sequence) in solution was also detected accurately by coupling to its complementary sequence probed on QDs-encoded beads through DNA hybridization assay. The resolution of this technology for encoded beads is resulted from two longer wavelength fluorescence identification signals (yellow and red fluorescent signals of QDs), and the third shorter wavelength fluorescence signal (green reporting signal of fluorescein isothiocyanate (FITC)) for the determination of reaction between probe and target. In experiment, because of QDs' unique optical character, only one excitation light source was needed to excite the QDs and probe dye FITC synchronously comparing with other flow cytometric assay technology. The results show that this technology has present excellent repeatability and good accuracy. It will become a promising multiple assay platform in various application fields after further improvement
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Flow cytometric characterization of cerebrospinal fluid cells.
de Graaf, Marieke T; de Jongste, Arjen H C; Kraan, Jaco; Boonstra, Joke G; Sillevis Smitt, Peter A E; Gratama, Jan W
2011-09-01
Flow cytometry facilitates the detection of a large spectrum of cellular characteristics on a per cell basis, determination of absolute cell numbers and detection of rare events with high sensitivity and specificity. White blood cell (WBC) counts in cerebrospinal fluid (CSF) are important for the diagnosis of many neurological disorders. WBC counting and differential can be performed by microscopy, hematology analyzers, or flow cytometry. Flow cytometry of CSF is increasingly being considered as the method of choice in patients suspected of leptomeningeal localization of hematological malignancies. Additionally, in several neuroinflammatory diseases such as multiple sclerosis and paraneoplastic neurological syndromes, flow cytometry is commonly performed to obtain insight into the immunopathogenesis of these diseases. Technically, the low cellularity of CSF samples, combined with the rapidly declining WBC viability, makes CSF flow cytometry challenging. Comparison of flow cytometry with microscopic and molecular techniques shows that each technique has its own advantages and is ideally combined. We expect that increasing the number of flow cytometric parameters that can be simultaneously studied within one sample, will further refine the information on CSF cell subsets in low-cellular CSF samples and enable to define cell populations more accurately. Copyright © 2011 International Clinical Cytometry Society.
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Multivariate analysis of flow cytometric data using decision trees.
Simon, Svenja; Guthke, Reinhard; Kamradt, Thomas; Frey, Oliver
2012-01-01
Characterization of the response of the host immune system is important in understanding the bidirectional interactions between the host and microbial pathogens. For research on the host site, flow cytometry has become one of the major tools in immunology. Advances in technology and reagents allow now the simultaneous assessment of multiple markers on a single cell level generating multidimensional data sets that require multivariate statistical analysis. We explored the explanatory power of the supervised machine learning method called "induction of decision trees" in flow cytometric data. In order to examine whether the production of a certain cytokine is depended on other cytokines, datasets from intracellular staining for six cytokines with complex patterns of co-expression were analyzed by induction of decision trees. After weighting the data according to their class probabilities, we created a total of 13,392 different decision trees for each given cytokine with different parameter settings. For a more realistic estimation of the decision trees' quality, we used stratified fivefold cross validation and chose the "best" tree according to a combination of different quality criteria. While some of the decision trees reflected previously known co-expression patterns, we found that the expression of some cytokines was not only dependent on the co-expression of others per se, but was also dependent on the intensity of expression. Thus, for the first time we successfully used induction of decision trees for the analysis of high dimensional flow cytometric data and demonstrated the feasibility of this method to reveal structural patterns in such data sets.
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Polymerization of N-(fluoro phenyl) maleimides
International Nuclear Information System (INIS)
Barrales-Rienda, J.M.; Ramos, J.G.; Chaves, M.S.
1979-01-01
Poly(N-aryl maleimide)s of characteristic structures have been synthesized and some of their physical properties studied. The polymerization of N-(fluoro phenyl) maleimides by free-radical initiation in bulk or in solution and by anionic catalyst have been studied to compare the characteristics of polymerization by γ-ray irradiation with that by free-radical initiation. The polymers were characterized by elemental analysis, intrinsic viscosity, spectroscopy (IR and NMR), programmed thermogravimetric analysis, and x-ray diffraction. Spectra of polymers prepared by radiation and anionic polymerization were nearly identical with those of polymers prepared by free-radical polymerization initiated by azobisisobutyronitrile in bulk or in solution and by the self-initiated thermal polymerization. A variety of reaction conditions were tried, but all attempts to change the molecular structure of the polymers were unsuccessful. Rates of thermal degradation for poly[N-(fluoro phenyl) maleimide]s have been analyzed by using a multiple-heating-rate procedure. Overall activation energy, order of reaction, and frequency factor have been evaluated. 6 figures, 8 tables
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Cluster Analysis of Flow Cytometric List Mode Data on a Personal Computer
Bakker Schut, Tom C.; Bakker schut, T.C.; de Grooth, B.G.; Greve, Jan
1993-01-01
A cluster analysis algorithm, dedicated to analysis of flow cytometric data is described. The algorithm is written in Pascal and implemented on an MS-DOS personal computer. It uses k-means, initialized with a large number of seed points, followed by a modified nearest neighbor technique to reduce
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A flow-cytometric NK-cytotoxicity assay adapted for use in rat repeated dose toxicity studies
International Nuclear Information System (INIS)
Marcusson-Staahl, Maritha; Cederbrant, Karin
2003-01-01
A recent regulatory document for immunotoxicity testing of new pharmaceutical drugs includes cytotoxic natural killer (NK)-cell function as a required parameter in repeated dose toxicity studies. The classical 51 Cr-release assay is the conventional test for cytotoxicity testing but several drawbacks with this assay has increased the demand for new reliable test systems. Here, we describe the optimisation of a flow-cytometric cytotoxicity assay especially adapted for regulatory rat studies in drug development. The test principle is based on target cell labelling with 5-(6)-carboxy-fluorescein succinimidyl ester (CFSE) and subsequent DNA-labelling with propidium iodide (PI) for identification of target cells with compromised cell membranes. The results are expressed as percentage of dead targets on a cell-to-cell basis. The final format of the assay includes 0.5 ml peripheral blood, 1.25x10 5 effector cells per sample, and collection of 500 target events by flow-cytometry. When NKR-P1+ cells were removed from the effector cell population by magnetic depletion the relative proportion decreased from 6 to 0.08%. The corresponding cytotoxic activity decreased from 68 to 8%. Also, the cytotoxic activity showed a significant and positive correlation with the proportion of NK-cells present in the effector cell suspension. Thus, the cytotoxicity measured is almost exclusively exerted by NK-cells. The current flow-cytometric test benefits from using peripheral blood as a source for effector cells since it will not conflict with the use of spleen for histopathological investigations in repeated dose toxicity studies. Additionally, since only a minimal number of effector cells are required per sample repeated testing of the same animal is enabled
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DEFF Research Database (Denmark)
Jensen, P O; Larsen, J K; Christensen, I J
1994-01-01
Bromodeoxyuridine (BrdUrd) labelled and unlabelled mitotic cells, respectively, can be discriminated from interphase cells using a new method, based on immunocytochemical staining of BrdUrd and flow cytometric four-parameter analysis of DNA content, BrdUrd incorporation, and forward and orthogonal...... light scatter. The method was optimized using the human leukemia cell lines HL-60 and K-562. Samples of 10(5) ethanol-fixed cells were treated with pepsin/HCl and stained as a nuclear suspension with anti-BrdUrd antibody, FITC-conjugated secondary antibody, and propidium iodide. Labelled mitoses could...
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DEFF Research Database (Denmark)
Jensen, P O; Larsen, J K; Christensen, I J
1994-01-01
Bromodeoxyuridine (BrdUrd) labelled and unlabelled mitotic cells, respectively, can be discriminated from interphase cells using a new method, based on immunocytochemical staining of BrdUrd and flow cytometric four-parameter analysis of DNA content, BrdUrd incorporation, and forward and orthogona...
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Ford, C H; Tsaltas, G C; Osborne, P A; Addetia, K
1996-03-01
A flow cytometric method of studying the internalization of a monoclonal antibody (Mab) directed against carcinoembryonic antigen (CEA) has been compared with Western blotting, using three human colonic cancer cell lines which express varying amounts of the target antigen. Cell samples incubated for increasing time intervals with fluoresceinated or unlabelled Mab were analyzed using flow cytometry or polyacrylamide gel electrophoresis and Western blotting. SDS/PAGE analysis of cytosolic and membrane components of solubilized cells from the cell lines provided evidence of non-degraded internalized anti-CEA Mab throughout seven half hour intervals, starting at 5 min. Internalized anti-CEA was detected in the case of high CEA expressing cell lines (LS174T, SKCO1). Very similar results were obtained with an anti-fluorescein flow cytometric assay. Given that these two methods consistently provided comparable results, use of flow cytometry for the detection of internalized antibody is suggested as a rapid alternative to most currently used methods for assessing antibody internalization. The question of the endocytic route followed by CEA-anti-CEA complexes was addressed by using hypertonic medium to block clathrin mediated endocytosis.
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A flow cytometric technique for quantification and differentiation of bacteria in bulk tank milk
DEFF Research Database (Denmark)
Holm, C.; Mathiasen, T.; Jespersen, Lene
2004-01-01
were defined: region 1 includes bacteria mainly associated with poor hygiene, region 2 includes psychrotrophic hygiene bacteria and region 3 includes bacteria mainly related to mastitis. The ability of the flow cytometric technique to predict the main cause of elevated bacterial counts on routine...
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Use of LysoTracker dyes: a flow cytometric study of autophagy.
Chikte, Shaheen; Panchal, Neelam; Warnes, Gary
2014-02-01
The flow cytometric use of LysoTracker dyes was employed to investigate the autophagic process and to compare this with the upregulation of autophagy marker, the microtubule-associated protein LC3B. Although the mechanism of action of LysoTracker dyes is not fully understood, they have been used in microscopy to image acidic spherical organelles, and their use in flow cytometry has not been thoroughly investigated in the study of autophagy. This investigation uses numerous autophagy-inducing agents including chloroquine (CQ), rapamycin, low serum (used to analyze patient cells as well as easier to use and significantly less costly. Copyright © 2013 International Society for Advancement of Cytometry.
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Photodynamic inactivation of foodborne bacteria by eosin Y.
Bonin, E; Dos Santos, A R; Fiori da Silva, A; Ribeiro, L H; Favero, M E; Campanerut-Sá, P A Z; de Freitas, C F; Caetano, W; Hioka, N; Mikcha, J M G
2018-03-25
The aim of this study was evaluate the effect of photodynamic inactivation mediated by eosin Y in Salmonella enterica serotype Typhimurium ATCC 14028, Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853, Staphylococcus aureus ATCC 25923 and Bacillus cereus ATCC 11778. Bacteria (10 7 CFU per ml) were incubated with eosin Y at concentrations ranging from 0·1 to 10 μmol l -1 , irradiated by green LED (λ max 490-570 nm) for 5, 10 and 15 min and the cellular viability was determined. Pseudomonas aeruginosa was completely inactivated when treated with 10 μmol l -1 eosin Y for 10 min. Treatments reduced B. cereus and Salm. Typhimurium counts to 2·7 log CFU per ml and 1·7 log CFU per ml, respectively. Escherichia coli counts were slightly reduced. Staphylococcus aureus presented the highest sensitivity, being completely inactivated by eosin Y at 5 μmol l -1 and 5 min of illumination. The reduction of cellular viability of photoinactivated Staph. aureus was also demonstrated by flow cytometry and morphological changes were observed by scanning electron microscopy. Eosin Y in combination with LED produced bacterial inactivation, being a potential candidate for photodynamic inactivation. This study evidenced the efficacy of photodynamic inactivation as a novel and promising alternative to bacterial control. © 2018 The Society for Applied Microbiology.
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Józwa, Wojciech; Czaczyk, Katarzyna
2012-04-02
Flow cytometry constitutes an alternative for traditional methods of microorganisms identification and analysis, including methods requiring cultivation step. It enables the detection of pathogens and other microorganisms contaminants without the need to culture microbial cells meaning that the sample (water, waste or food e.g. milk, wine, beer) may be analysed directly. This leads to a significant reduction of time required for analysis allowing monitoring of production processes and immediate reaction in case of contamination or any disruption occurs. Apart from the analysis of raw materials or products on different stages of manufacturing process, the flow cytometry seems to constitute an ideal tool for the assessment of microbial contamination on the surface of technological lines. In the present work samples comprising smears from 3 different surfaces of technological lines from fruit and vegetable processing company from Greater Poland were analysed directly with flow cytometer. The measured parameters were forward and side scatter of laser light signals allowing the estimation of microbial cell contents in each sample. Flow cytometric analysis of the surface of food industry production lines enable the preliminary evaluation of microbial contamination within few minutes from the moment of sample arrival without the need of sample pretreatment. The presented method of fl ow cytometric initial evaluation of microbial state of food industry technological lines demonstrated its potential for developing a robust, routine method for the rapid and labor-saving detection of microbial contamination in food industry.
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Jenkins, J A; Draugelis-Dale, R O; Pinkney, A E; Iwanowicz, L R; Blazer, V S
2015-03-15
Declining harvests of yellow perch, Perca flavescens, in urbanized watersheds of Chesapeake Bay have prompted investigations of their reproductive fitness. The purpose of this study was to establish a flow cytometric technique for DNA analysis of fixed samples sent from the field to provide reliable gamete quality measurements. Similar to the sperm chromatin structure assay, measures were made on the susceptibility of nuclear DNA to acid-induced denaturation, but used fixed rather than live or thawed cells. Nuclei were best exposed to the acid treatment for 1 minute at 37 °C followed by the addition of cold (4 °C) propidium iodide staining solution before flow cytometry. The rationale for protocol development is presented graphically through cytograms. Field results collected in 2008 and 2009 revealed DNA fragmentation up to 14.5%. In 2008, DNA fragmentation from the more urbanized watersheds was significantly greater than from reference sites (P = 0.026) and in 2009, higher percentages of haploid testicular cells were noted from the less urbanized watersheds (P = 0.032) indicating better reproductive condition at sites with less urbanization. For both years, total and progressive live sperm motilities by computer-assisted sperm motion analysis ranged from 19.1% to 76.5%, being significantly higher at the less urbanized sites (P < 0.05). This flow cytometric method takes advantage of the propensity of fragmented DNA to be denatured under standard conditions, or 1 minute at 37 °C with 10% buffered formalin-fixed cells. The study of fixed sperm makes possible the restrospective investigation of germplasm fragmentation, spermatogenic ploidy patterns, and chromatin compaction levels from samples translocated over distance and time. The protocol provides an approach that can be modified for other species across taxa. Published by Elsevier Inc.
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Gifford, Carrie E; Weingartner, Elizabeth; Villanueva, Joyce; Johnson, Judith; Zhang, Kejian; Filipovich, Alexandra H; Bleesing, Jack J; Marsh, Rebecca A
2014-07-01
X-linked lymphoproliferative disease is caused by mutations in two genes, SH2D1A and XIAP/BIRC4. Flow cytometric methods have been developed to detect the gene products, SAP and XIAP. However, there is no literature describing the accuracy of flow cytometric screening performed in a clinical lab setting. We reviewed the clinical flow cytometric testing results for 656 SAP and 586 XIAP samples tested during a 3-year period. Genetic testing was clinically performed as directed by the managing physician in 137 SAP (21%) and 115 XIAP (20%) samples. We included these samples for analyses of flow cytometric test accuracy. SH2D1A mutations were detected in 15/137 samples. SAP expression was low in 13/15 (sensitivity 87%, CI 61-97%). Of the 122 samples with normal sequencing, SAP was normal in 109 (specificity 89%, CI 82-94%). The positive predictive values (PPVs) and the negative predictive values (NPVs) were 50% and 98%, respectively. XIAP/BIRC4 mutations were detected in 19/115 samples. XIAP expression was low in 18/19 (sensitivity 95%, CI 73-100%). Of the 96 samples with normal sequencing, 59 had normal XIAP expression (specificity 61%, CI 51-71%). The PPVs and NPVs were 33% and 98%, respectively. Receiver-operating characteristic analysis was able to improve the specificity to 75%. Clinical flow cytometric screening tests for SAP and XIAP deficiencies offer good sensitivity and specificity for detecting genetic mutations, and are characterized by high NPVs. We recommend these tests for patients suspected of having X-linked lymphoproliferative disease type 1 (XLP1) or XLP2. © 2014 Clinical Cytometry Society.
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A flow-cytometric gram-staining technique for milk-associated bacteria.
Holm, Claus; Jespersen, Lene
2003-05-01
A Gram-staining technique combining staining with two fluorescent stains, Oregon Green-conjugated wheat germ agglutinin (WGA) and hexidium iodide (HI) followed by flow-cytometric detection is described. WGA stains gram-positive bacteria while HI binds to the DNA of all bacteria after permeabilization by EDTA and incubation at 50 degrees C for 15 min. For WGA to bind to gram-positive bacteria, a 3 M potassium chloride solution was found to give the highest fluorescence intensity. A total of 12 strains representing some of the predominant bacterial species in bulk tank milk and mixtures of these were stained and analyzed by flow cytometry. Overall, the staining method showed a clear differentiation between gram-positive and gram-negative bacterial populations. For stationary-stage cultures of seven gram-positive bacteria and five gram-negative bacteria, an average of 99% of the cells were correctly interpreted. The method was only slightly influenced by the growth phase of the bacteria or conditions such as freezing at -18 degrees C for 24 h. For any of these conditions, an average of at least 95% of the cells were correctly interpreted. When stationary-stage cultures were stored at 5 degrees C for 14 days, an average of 86% of the cells were correctly interpreted. The Gram-staining technique was applied to the flow cytometry analysis of bulk tank milk inoculated with Staphylococcus aureus and Escherichia coli. These results demonstrate that the technique is suitable for analyzing milk samples without precultivation.
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DEFF Research Database (Denmark)
Vindeløv, L L; Hansen, H H; Christensen, I J
1980-01-01
Flow-cytometric DNA analysis yields information on ploidy and proliferative characteristics of a cell population. The analysis was implemented on small-cell anaplastic carcinoma of the lung using a rapid detergent technique for the preparation of fine-needle aspirates for DNA determination and a ...
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The cytometric future: it ain't necessarily flow!
Shapiro, Howard M
2011-01-01
Initial approaches to cytometry for classifying and characterizing cells were based on microscopy; it was necessary to collect relatively high-resolution images of cells because only a few specific reagents usable for cell identification were available. Although flow cytometry, now the dominant cytometric technology, typically utilizes lenses similar to microscope lenses for light collection, improved, more quantitative reagents allow the necessary information to be acquired in the form of whole-cell measurements of the intensities of light transmission, scattering, and/or fluorescence.Much of the cost and complexity of both automated microscopes and flow cytometers arises from the necessity for them to measure one cell at a time. Recent developments in digital camera technology now offer an alternative in which one or more low-magnification, low-resolution images are made of a wide field containing many cells, using inexpensive light-emitting diodes (LEDs) for illumination. Minimalist widefield imaging cytometers can provide a smaller, less complex, and substantially less expensive alternative to flow cytometry, critical in systems intended for in resource-poor areas. Minimalism is, likewise, a good philosophy in developing instrumentation and methodology for both clinical and large-scale research use; it simplifies quality assurance and compliance with regulatory requirements, as well as reduces capital outlays, material costs, and personnel training requirements. Also, importantly, it yields "greener" technology.
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International Nuclear Information System (INIS)
Hoshino, S.; Oshimi, K.; Tsudo, M.; Miyasaka, M.; Teramura, M.; Masuda, M.; Motoji, T.; Mizoguchi, H.
1990-01-01
We analyzed the expression of the interleukin-2 receptor (IL-2R) beta chain (p70-75) on various leukemic cells from 44 patients by flow cytometric analysis using the IL-2R beta chain-specific monoclonal antibody, designated Mik-beta 1. Flow cytometric analysis demonstrated the expression of the IL-2R beta chain on granular lymphocytes (GLs) from all eight patients with granular lymphocyte proliferative disorders (GLPDs), on adult T-cell leukemia (ATL) cells from all three patients with ATL, and on T-cell acute lymphoblastic leukemia (T-ALL) cells from one of three patients with T-ALL. Although GLs from all the GLPD patients expressed the IL-2R beta chain alone and not the IL-2R alpha chain (Tac-antigen: p55), ATL and T-ALL cells expressing the beta chain coexpressed the alpha chain. In two of seven patients with common ALL (cALL) and in both patients with B-cell chronic lymphocytic leukemia, the leukemic cells expressed the alpha chain alone. Neither the alpha chain nor the beta chain was expressed on leukemic cells from the remaining 28 patients, including all 18 patients with acute nonlymphocytic leukemia, five of seven patients with cALL, all three patients with multiple myeloma, and two of three patients with T-ALL. These results indicate that three different forms of IL-2R chain expression exist on leukemic cells: the alpha chain alone; the beta chain alone; and both the alpha and beta chains. To examine whether the results obtained by flow cytometric analysis actually reflect functional aspects of the expressed IL-2Rs, we studied the specific binding of 125I-labeled IL-2 (125I-IL-2) to leukemic cells in 18 of the 44 patients. In addition, we performed 125I-IL-2 crosslinking studies in seven patients. The results of IL-2R expression of both 125I-IL-2 binding assay and crosslinking studies were in agreement with those obtained by flow cytometric analysis
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Blacking, T M; Waterfall, M; Samuel, K; Argyle, D J
2012-12-01
The cancer stem cell (CSC) hypothesis proposes that tumour growth is maintained by a distinct subpopulation of 'CSC'. This study applied flow cytometric methods, reported to detect CSC in both primary and cultured cancer cells of other species, to identify candidate canine subpopulations. Cell lines representing diverse canine malignancies, and cells derived from spontaneous canine tumours, were evaluated for expression of stem cell-associated surface markers (CD34, CD44, CD117 and CD133) and functional properties [Hoecsht 33342 efflux, aldehyde dehydrogenase (ALDH) activity]. No discrete marker-defined subsets were identified within established cell lines; cells derived directly from spontaneous tumours demonstrated more heterogeneity, although this diminished upon in vitro culture. Functional assays produced variable results, suggesting context-dependency. Flow cytometric methods may be adopted to identify putative canine CSC. Whilst cell lines are valuable in assay development, primary cells may provide a more rewarding model for studying tumour heterogeneity in the context of CSC. However, it will be essential to fully characterize any candidate subpopulations to ensure that they meet CSC criteria. © 2011 Blackwell Publishing Ltd.
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Bommannan, K; Sachdeva, M U S; Gupta, M; Bose, P; Kumar, N; Sharma, P; Naseem, S; Ahluwalia, J; Das, R; Varma, N
2016-10-01
A good bone marrow (BM) sample is essential in evaluating many hematologic disorders. An unsuccessful BM aspiration (BMA) procedure precludes a successful flow cytometric immunophenotyping (FCI) in most hematologic malignancies. Apart from FCI, most ancillary diagnostic techniques in hematology are less informative. We describe the feasibility of FCI in vortex-dislodged cell preparation obtained from unfixed trephine biopsy (TB) specimens. In pancytopenic patients and dry tap cases, routine diagnostic BMA and TB samples were complemented by additional trephine biopsies. These supplementary cores were immediately transferred into sterile tubes filled with phosphate-buffered saline, vortexed, and centrifuged. The cell pellet obtained was used for flow cytometric immunophenotyping. Of 7955 BMAs performed in 42 months, 34 dry tap cases were eligible for the study. Vortexing rendered a cell pellet in 94% of the cases (32 of 34), and FCI rendered a rapid diagnosis in 100% of the cases (32 of 32) where cell pellets were available. We describe an efficient procedure which could be effectively utilized in resource-limited centers and reduce the frequency of repeat BMA procedures. © 2016 John Wiley & Sons Ltd.
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International Nuclear Information System (INIS)
Jacobs, R.M.; Boyce, J.T.; Kociba, G.J.
1986-01-01
This study demonstrates the potential usefulness of a flow cytometric technique to measure platelet survival time in cats utilizing autologous platelets labeled in vitro with fluorescein isothiocyanate (FITC). When compared with a 51Cr method, no significant differences in estimated survival times were found. Both the 51Cr and FITC-labeling procedures induced similar changes in platelet shape and collagen-induced aggregation. Platelets labeled with FITC had significantly greater volumes compared with those of glutaraldehyde-fixed platelets. These changes were primarily related to the platelet centrifugation and washing procedures rather than the labels themselves. This novel technique potentially has wide applicability to cell circulation time studies as flow cytometry equipment becomes more readily available. Problems with the technique are discussed. In a preliminary study of the platelet survival time in feline leukemia virus (FeLV)-infected cats, two of three cats had significantly reduced survival times using both flow cytometric and radioisotopic methods. These data suggest increased platelet turnover in FeLV-infected cats
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Energy Technology Data Exchange (ETDEWEB)
Jacobs, R.M.; Boyce, J.T.; Kociba, G.J.
1986-01-01
This study demonstrates the potential usefulness of a flow cytometric technique to measure platelet survival time in cats utilizing autologous platelets labeled in vitro with fluorescein isothiocyanate (FITC). When compared with a 51Cr method, no significant differences in estimated survival times were found. Both the 51Cr and FITC-labeling procedures induced similar changes in platelet shape and collagen-induced aggregation. Platelets labeled with FITC had significantly greater volumes compared with those of glutaraldehyde-fixed platelets. These changes were primarily related to the platelet centrifugation and washing procedures rather than the labels themselves. This novel technique potentially has wide applicability to cell circulation time studies as flow cytometry equipment becomes more readily available. Problems with the technique are discussed. In a preliminary study of the platelet survival time in feline leukemia virus (FeLV)-infected cats, two of three cats had significantly reduced survival times using both flow cytometric and radioisotopic methods. These data suggest increased platelet turnover in FeLV-infected cats.
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Cytometric analysis of mammalian sperm for induced morphologic and DNA content errors
International Nuclear Information System (INIS)
Pinkel, D.
1983-01-01
Some flow-cytometric and image analysis procedures under development for quantitative analysis of sperm morphology are reviewed. The results of flow-cytometric DNA-content measurements on sperm from radiation exposed mice are also summarized, the results related to the available cytological information, and their potential dosimetric sensitivity discussed
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Flow cytometric of reticulocytes quantification: radio-induction medullary aplasia application
International Nuclear Information System (INIS)
Dubner, D.; Perez, M.; Gisone, P.
1996-01-01
Flow cytometric reticulocyte quantification was assayed in ten patients undergoing bone marrow transplantation (BMT) with previous conditioning by chemotherapy and total body irradiation. A reticulocyte maturity index (RMI) was determined taking into account the RNA content. With the aim of testing the utility of RMI as an early predictor of functional recovery in marrow aplasia, other hematological indicators as neutrophils count were comparatively evaluated. Mean time elapsed between BMT and engraftment evidence by RMI was 17,6 days. In six patients the RMI was the earliest indicator of functional recovery. The applicability of this assay in the pursuit of radioinduced bone marrow aplasia is discussed. (authors). 4 refs., 4 figs., 2 tabs
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Jacques, Nathalie; Vimond, Nadege; Conforti, Rosa; Griscelli, Franck; Lecluse, Yann; Laplanche, Agnes; Malka, David; Vielh, Philippe; Farace, Françoise
2008-09-15
Circulating endothelial cells (CEC) are currently proposed as a potential biomarker for measuring the impact of anti-angiogenic treatments in cancer. However, the lack of consensus on the appropriate method of CEC measurement has led to conflicting data in cancer patients. A validated assay adapted for evaluating the clinical utility of CEC in large cohorts of patients undergoing anti-angiogenic treatments is needed. We developed a four-color flow cytometric assay to measure CEC as CD31(+), CD146(+), CD45(-), 7-amino-actinomycin-D (7AAD)(-) events in whole blood. The distinctive features of the assay are: (1) staining of 1 ml whole blood, (2) use of a whole blood IgPE control to measure accurately background noise, (3) accumulation of a large number of events (almost 5 10(6)) to ensure statistical analysis, and (4) use of 10 microm fluorescent microbeads to evaluate the event size. Assay reproducibility was determined in duplicate aliquots of samples drawn from 20 metastatic cancer patients. Assay linearity was tested by spiking whole blood with low numbers of HUVEC. Five-color flow cytometric experiments with CD144 were performed to confirm the endothelial origin of the cells. CEC were measured in 20 healthy individuals and 125 patients with metastatic cancer. Reproducibility was good between duplicate aliquots (r(2)=0.948, mean difference between duplicates of 0.86 CEC/ml). Detected HUVEC correlated with spiked HUVEC (r(2)=0.916, mean recovery of 100.3%). Co-staining of CD31, CD146 and CD144 confirmed the endothelial nature of cells identified as CEC. Median CEC levels were 6.5/ml (range, 0-15) in healthy individuals and 15.0/ml (range, 0-179) in patients with metastatic carcinoma (p<0.001). The assay proposed here allows reproducible and sensitive measurement of CEC by flow cytometry and could help evaluate CEC as biomarkers of anti-angiogenic therapies in large cohorts of patients.
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International Nuclear Information System (INIS)
Zak, K.P.; Gruzov, M.A.; Bolshova, B.V.; Afanasyeva, V.V.; Shlyakhovenko, V.S.; Vishnevskaya, O.A.; Tronko, N.D.
1996-01-01
It hag been carded out a light, election microscopic and flow cytometric study of blood leukocyte of children with malignant tumors (papillary carcinoma) of thyroid gland who were living at the moment of the accident near Chernobyl. The results obtained point out the presence of some disturbances of immune status of these children
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International Nuclear Information System (INIS)
McCreedy, C.D.; Jagoe, C.H.; Brisbin, I.L. Jr.; Wentworth, R.W.; Dallas, C.E.
1995-01-01
Clinical technologies, such as flow cytometry, are increasingly adopted by environmental toxicologists to identify resource damage associated with exposure to xenobiotics. One application of flow cytometry allows the rapid determination of the DNA content of large numbers of individual cells, and can be used to detect aneuploidy or other genetic abnormalities. The laboratory has used this methodology in studies of genetic toxicology of fish, birds, arid mammals exposed to organic pollutants, metals and radionuclides, However, without appropriate quality controls, false positive results and other artifacts can arise from sample handling and preparations, inter and intra-individual variations, instrument noise and other sources. The authors describe the routine measures this laboratory employs to maintain quality control of genomic DNA analysis, including the control of staining conditions, machine standardization, pulse-width doublet discrimination, and, in particular, the use of internal controls and the use of time as a cytometric parameter. Neglect of these controls can produce erroneous results, leading to conclusions of genetic abnormalities when none are present. Conversely, attention to these controls, routinely used in clinical settings, facilitates the interpretation of flow cytometric data and allows the application of this sensitive indicator of genotoxic effects to a variety of environmental problems
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Flow Cytometric Analysis of T, B, and NK Cells Antigens in Patients with Mycosis Fungoides
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Serkan Yazıcı
2015-01-01
Full Text Available We retrospectively analyzed the clinicopathological correlation and prognostic value of cell surface antigens expressed by peripheral blood mononuclear cells in patients with mycosis fungoides (MF. 121 consecutive MF patients were included in this study. All patients had peripheral blood flow cytometry as part of their first visit. TNMB and histopathological staging of the cases were retrospectively performed in accordance with International Society for Cutaneous Lymphomas/European Organization of Research and Treatment of Cancer (ISCL/EORTC criteria at the time of flow cytometry sampling. To determine prognostic value of cell surface antigens, cases were divided into two groups as stable and progressive disease. 17 flow cytometric analyses of 17 parapsoriasis (PP and 11 analyses of 11 benign erythrodermic patients were included as control groups. Fluorescent labeled monoclonal antibodies were used to detect cell surface antigens: T cells (CD3+, CD4+, CD8+, TCRαβ+, TCRγδ+, CD7+, CD4+CD7+, CD4+CD7−, and CD71+, B cells (HLA-DR+, CD19+, and HLA-DR+CD19+, NKT cells (CD3+CD16+CD56+, and NK cells (CD3−CD16+CD56+. The mean value of all cell surface antigens was not statistically significant between parapsoriasis and MF groups. Along with an increase in cases of MF stage statistically significant difference was found between the mean values of cell surface antigens. Flow cytometric analysis of peripheral blood cell surface antigens in patients with mycosis fungoides may contribute to predicting disease stage and progression.
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Bence György
Full Text Available INTRODUCTION: Microvesicles (MVs, earlier referred to as microparticles, represent a major type of extracellular vesicles currently considered as novel biomarkers in various clinical settings such as autoimmune disorders. However, the analysis of MVs in body fluids has not been fully standardized yet, and there are numerous pitfalls that hinder the correct assessment of these structures. METHODS: In this study, we analyzed synovial fluid (SF samples of patients with osteoarthritis (OA, rheumatoid arthritis (RA and juvenile idiopathic arthritis (JIA. To assess factors that may confound MV detection in joint diseases, we used electron microscopy (EM, Nanoparticle Tracking Analysis (NTA and mass spectrometry (MS. For flow cytometry, a method commonly used for phenotyping and enumeration of MVs, we combined recent advances in the field, and used a novel approach of differential detergent lysis for the exclusion of MV-mimicking non-vesicular signals. RESULTS: EM and NTA showed that substantial amounts of particles other than MVs were present in SF samples. Beyond known MV-associated proteins, MS analysis also revealed abundant plasma- and immune complex-related proteins in MV preparations. Applying improved flow cytometric analysis, we demonstrate for the first time that CD3(+ and CD8(+ T-cell derived SF MVs are highly elevated in patients with RA compared to OA patients (p=0.027 and p=0.009, respectively, after Bonferroni corrections. In JIA, we identified reduced numbers of B cell-derived MVs (p=0.009, after Bonferroni correction. CONCLUSIONS: Our results suggest that improved flow cytometric assessment of MVs facilitates the detection of previously unrecognized disease-associated vesicular signatures.
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Rose, Jonathan A; Wanner, Nicholas; Cheong, Hoi I; Queisser, Kimberly; Barrett, Patrick; Park, Margaret; Hite, Corrine; Naga Prasad, Sathyamangla V; Erzurum, Serpil; Asosingh, Kewal
2016-01-01
Pulmonary arterial hypertension (PAH) is a heterogeneous disease characterized by severe angiogenic remodeling of the pulmonary artery wall and right ventricular hypertrophy. Thus, there is an increasing need for novel biomarkers to dissect disease heterogeneity, and predict treatment response. Although β-adrenergic receptor (βAR) dysfunction is well documented in left heart disease while endothelial cell-derived microparticles (Ec-MPs) are established biomarkers of angiogenic remodeling, methods for easy large clinical cohort analysis of these biomarkers are currently absent. Here we describe flow cytometric methods for quantification of βAR density on circulating white blood cells (WBC) and Ec-MPs in urine samples that can be used as potential biomarkers of right heart failure in PAH. Biotinylated β-blocker alprenolol was synthesized and validated as a βAR specific probe that was combined with immunophenotyping to quantify βAR density in circulating WBC subsets. Ec-MPs obtained from urine samples were stained for annexin-V and CD144, and analyzed by a micro flow cytometer. Flow cytometric detection of alprenolol showed that βAR density was decreased in most WBC subsets in PAH samples compared to healthy controls. Ec-MPs in urine was increased in PAH compared to controls. Furthermore, there was a direct correlation between Ec-MPs and Tricuspid annular plane systolic excursion (TAPSE) in PAH patients. Therefore, flow cytometric quantification of peripheral blood cell βAR density and urinary Ec-MPs may be useful as potential biomarkers of right ventricular function in PAH.
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Sadhu, Abhishek; Bhadra, Sreetama; Bandyopadhyay, Maumita
2016-11-01
Cytological parameters such as chromosome numbers and genome sizes of plants are used routinely for studying evolutionary aspects of polyploid plants. Members of Zingiberaceae show a wide range of inter- and intrageneric variation in their reproductive habits and ploidy levels. Conventional cytological study in this group of plants is severely hampered by the presence of diverse secondary metabolites, which also affect their genome size estimation using flow cytometry. None of the several nuclei isolation buffers used in flow cytometry could be used very successfully for members of Zingiberaceae to isolate good quality nuclei from both shoot and root tissues. The competency of eight nuclei isolation buffers was compared with a newly formulated buffer, MB01, in six different genera of Zingiberaceae based on the fluorescence intensity of propidium iodide-stained nuclei using flow cytometric parameters, namely coefficient of variation of the G 0 /G 1 peak, debris factor and nuclei yield factor. Isolated nuclei were studied using fluorescence microscopy and bio-scanning electron microscopy to analyse stain-nuclei interaction and nuclei topology, respectively. Genome contents of 21 species belonging to these six genera were determined using MB01. Flow cytometric parameters showed significant differences among the analysed buffers. MB01 exhibited the best combination of analysed parameters; photomicrographs obtained from fluorescence and electron microscopy supported the superiority of MB01 buffer over other buffers. Among the 21 species studied, nuclear DNA contents of 14 species are reported for the first time. Results of the present study substantiate the enhanced efficacy of MB01, compared to other buffers tested, in the generation of acceptable cytograms from all species of Zingiberaceae studied. Our study facilitates new ways of sample preparation for further flow cytometric analysis of genome size of other members belonging to this highly complex polyploid family
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Flow cytometric life cycle analysis in cellular radiation biology
International Nuclear Information System (INIS)
Wood, J.C.S.
1982-01-01
Three approaches to flow cytometric histogram analysis were developed: (1) differential histogram analysis, (2) DNA histogram analysis, and (3) multiparameter data analysis. These techniques were applied to an important unresolved problem in radiation biology. The initial responses to irradiation of a mammalian cell which occur during the first two cell cycles following the irradiation are of considerable interest to the radiation biologist. During the first two post-irradiation cell cycles, cells which ultimately will survive repair radiation-induced damage, while some cells begin to express some of the radiation-induced nuclear and chomatin damage. Caffeine- and thymidine-treated, and untreated gamma-irradiated cell populations were studied with respect to the radiation-induced G2 delay, deficient DNA synthesis, and the appearance of cells with abnormal DNA contents. It is hypothesized that the measured deficiency in DNA synthesis observed in the first post-irradiation cell cycle may be a result of daughter cells from abnormal first post-irradiation mitoses
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M.O. de Jong (Marg); H. Rozemuller (Henk); J.G.J. Bauman (J. G J); J.W.M. Visser (Jan)
1995-01-01
textabstractThe main prerequisites for the use of biotinylated ligands to study the expression of growth factor receptors on heterogeneous cell populations, such as peripheral blood or bone marrow, by flow cytometric methods, are that the biotinylated ligand retains its binding ability and that
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Carson, Christine F; Inglis, Timothy JJ
2018-01-01
This study investigated aerosolized viable bacteria in a university research laboratory during operation of an acoustic-assisted flow cytometer for antimicrobial susceptibility testing by sampling room air before, during and after flow cytometer use. The aim was to assess the risk associated with use of an acoustic-assisted flow cytometer analyzing unfixed bacterial suspensions. Air sampling in a nearby clinical laboratory was conducted during the same period to provide context for the existing background of microorganisms that would be detected in the air. The three species of bacteria undergoing analysis by flow cytometer in the research laboratory were Klebsiella pneumoniae, Burkholderia thailandensis and Streptococcus pneumoniae. None of these was detected from multiple 1000 L air samples acquired in the research laboratory environment. The main cultured bacteria in both locations were skin commensal and environmental bacteria, presumed to have been disturbed or dispersed in laboratory air by personnel movements during routine laboratory activities. The concentrations of bacteria detected in research laboratory air samples were reduced after interventional cleaning measures were introduced and were lower than those in the diagnostic clinical microbiology laboratory. We conclude that our flow cytometric analyses of unfixed suspensions of K. pneumoniae, B. thailandensis and S. pneumoniae do not pose a risk to cytometer operators or other personnel in the laboratory but caution against extrapolation of our results to other bacteria and/or different flow cytometric experimental procedures. PMID:29608197
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Karyological and flow cytometric evidence of triploid specimens in Bufo viridis (Amphibia Anura
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D Cavallo
2010-01-01
Full Text Available Karyological and flow cytometric (FCM analyses were performed on a group of 14 green toads of the Bufo viridis species from seven Eurasian populations. Both approaches gave concordant results concerning the DNA ploidy level. All the populations examined were represented exclusively by diploid or tetraploid specimens, except one, where triploids were found. Results evidenced an interpopulation variability in DNA content against the same ploidy level, as well as an unusually high number of triploids in a particular reproductive place. The origin of polyploidy and the presence and persistence of a high number of triploids in a particular population are discussed.
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Directory of Open Access Journals (Sweden)
Jonathan A Rose
Full Text Available Pulmonary arterial hypertension (PAH is a heterogeneous disease characterized by severe angiogenic remodeling of the pulmonary artery wall and right ventricular hypertrophy. Thus, there is an increasing need for novel biomarkers to dissect disease heterogeneity, and predict treatment response. Although β-adrenergic receptor (βAR dysfunction is well documented in left heart disease while endothelial cell-derived microparticles (Ec-MPs are established biomarkers of angiogenic remodeling, methods for easy large clinical cohort analysis of these biomarkers are currently absent. Here we describe flow cytometric methods for quantification of βAR density on circulating white blood cells (WBC and Ec-MPs in urine samples that can be used as potential biomarkers of right heart failure in PAH. Biotinylated β-blocker alprenolol was synthesized and validated as a βAR specific probe that was combined with immunophenotyping to quantify βAR density in circulating WBC subsets. Ec-MPs obtained from urine samples were stained for annexin-V and CD144, and analyzed by a micro flow cytometer. Flow cytometric detection of alprenolol showed that βAR density was decreased in most WBC subsets in PAH samples compared to healthy controls. Ec-MPs in urine was increased in PAH compared to controls. Furthermore, there was a direct correlation between Ec-MPs and Tricuspid annular plane systolic excursion (TAPSE in PAH patients. Therefore, flow cytometric quantification of peripheral blood cell βAR density and urinary Ec-MPs may be useful as potential biomarkers of right ventricular function in PAH.
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Flow cytometric and morphological analyses of Pinus pinaster somatic embryogenesis.
Marum, Liliana; Loureiro, João; Rodriguez, Eleazar; Santos, Conceição; Oliveira, M Margarida; Miguel, Célia
2009-09-25
An approach combining morphological profiling and flow cytometric analysis was used to assess genetic stability during the several steps of somatic embryogenesis in Pinus pinaster. Embryogenic cell lines of P. pinaster were established from immature zygotic embryos excised from seeds obtained from open-pollinated trees. During the maturation stage, phenotype of somatic embryos was characterized as being either normal or abnormal. Based upon the prevalent morphological traits, different types of abnormal embryos underwent further classification and quantification. Nuclear DNA content of maritime pine using the zygotic embryos was estimated to be 57.04 pg/2C, using propidium iodide flow cytometry. According to the same methodology, no significant differences (P< or =0.01) in DNA ploidy were detected among the most frequently observed abnormal phenotypes, embryogenic cell lines, zygotic and normal somatic embryos, and somatic embryogenesis-derived plantlets. Although the differences in DNA ploidy level do not exclude the occurrence of a low level of aneuploidy, the results obtained point to the absence of major changes in ploidy level during the somatic embryogenesis process of this economically important species. Therefore, our primary goal of true-to-typeness was assured at this level.
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A novel flow cytometric assay for measurement of In Vivo pulmonary neutrophil phagocytosis
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Gentry-Nielsen Martha J
2006-07-01
Full Text Available Abstract Background Phagocytosis assays are traditionally performed in vitro using polymorphonuclear leukocytes (PMNs isolated from peripheral blood or the peritoneum and heat-killed, pre-opsonized organisms. These assays may not adequately mimic the environment within the infected lung. Our laboratory therefore has developed a flow cytometric in vivo phagocytosis assay that enables quantification of PMN phagocytosis of viable bacteria within the lungs of rats. In these studies, rats are injected transtracheally with lipopolysaccharide (LPS to recruit PMNs to their lungs. They are then infected with live 5(-and 6 carboxyfluorescein diacetate succinimidyl ester (CFDA/SE labeled type 3 Streptococcus pneumoniae. Bronchoalveolar lavage is performed and resident alveolar macrophages and recruited PMNs are labeled with monoclonal antibodies specific for surface epitopes on each cell type. Three color flow cytometry is utilized to identify the cell types, quantify recruitment, and determine uptake of the labeled bacteria. Results The viability of the alveolar macrophages and PMNs isolated from the lavage fluid was >95%. The values of the percentage of PMNs in the lavage fluid as well as the percentage of PMNs associated with CFSE-labeled S. pneumoniae as measured through flow cytometry showed a high degree of correlation with the results from manual counting of cytospin slides. Conclusion This assay is suitable for measuring bacterial uptake within the infected lung. It can be adapted for use with other organisms and/or animal model systems.
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Novel ?-cyclodextrin?eosin conjugates
Benkovics, G?bor; Afonso, Damien; Darcsi, Andr?s; B?ni, Szabolcs; Conoci, Sabrina; Fenyvesi, ?va; Szente, Lajos; Malanga, Milo; Sortino, Salvatore
2017-01-01
Eosin B (EoB) and eosin Y (EoY), two xanthene dye derivatives with photosensitizing ability were prepared in high purity through an improved synthetic route. The dyes were grafted to a 6-monoamino-β-cyclodextrin scaffold under mild reaction conditions through a stable amide linkage using the coupling agent 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium chloride. The molecular conjugates, well soluble in aqueous medium, were extensively characterized by 1D and 2D NMR spectroscopy an...
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Rainer Kimmig
2001-01-01
Full Text Available Flow cytometric assessment of DNA‐ploidy and S‐phase fraction in malignant tumors is compromised by the heterogeneity of cell subpopulations derived from the malignant and surrounding connective tissue, e.g., tumor, stromal and inflammatory cells. To evaluate the effect on quality of DNA cell cycle analysis and determination of DNA ploidy, cytokeratin labeling of epithelial cells was used for tumor cell enrichment in breast, ovarian, cervical and endometrial cancer prior to DNA analysis. In a prospective study, tumor cell subpopulations of 620 malignant tumors were labeled by a FITC‐conjugated cytokeratin antibody (CK 5, 6, CK18 and CK 5, 6, 8 and CK 17, respectively prior to flow cytometric cell cycle analysis. Compared to total cell analysis, detection rate of DNA‐aneuploid tumors following cytokeratin labeling was increased from 62% to 76.5% in breast cancer, from 68% to 77% in ovarian cancer, from 60% to 80% in cervical cancer and from 30% to 53% in endometrial cancer. Predominantly in DNA‐diploid tumors, a significantly improved detection of S‐phase fraction of the tumor cells was shown due to the elimination of contaminating nonproliferating “normal cells”. S‐phase fraction following tumor cell enrichment was increased by 10% (mean following cytokeratin staining in ovarian and endometrial cancer, by 30% in breast cancer and even by 70% in cervical cancer compared to total cell analysis. Thus, diagnostic accuracy of DNA‐analysis was enhanced by cytokeratin labeling of tumor cells for all tumor entities investigated.
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Flow cytometric DNA analysis of ducks accumulating 137Cs on a reactor reservoir
International Nuclear Information System (INIS)
George, L.S.; Dallas, C.E.; Brisbin, I.L. Jr.; Evans, D.L.
1991-01-01
The objective of this study was to detect red blood cell (rbc) DNA abnormalities in male, game-farm mallard ducks as they ranged freely and accumulated 137Cs (radiocesium) from an abandoned nuclear reactor cooling reservoir. Prior to release, the ducks were tamed to enable recapture at will. Flow cytometric measurements conducted at intervals during the first year of exposure yielded cell cycle percentages of DNA (G0/G1, S, G2 + M phases) of rbc, as well as coefficients of variation (CV) in the G0/G1 phase. DNA histograms of exposed ducks were compared with two sets of controls which were maintained 30 and 150 miles from the study site. 137Cs live wholebody burdens were also measured in these animals in a parallel kinetics study, and an approximate steady-state equilibrium was attained after about 8 months. DNA histograms from 2 of the 14 contaminated ducks revealed DNA aneuploid-like patterns after 9 months exposure. These two ducks were removed from the experiment at this time, and when sampled again 1 month later, one continued to exhibit DNA aneuploidy. None of the control DNA histograms demonstrated DNA aneuploid-like patterns. There were no significant differences in cell cycle percentages at any time point between control and exposed animals. A significant increase in CV was observed at 9 months exposure, but after removal of the two ducks with DNA aneuploidy, no significant difference was detected in the group monitored after 12 months exposure. An increased variation in the DNA and DNA aneuploidy could, therefore, be detected in duck rbc using flow cytometric analysis, with the onset of these effects being related to the attainment of maximal levels of 137Cs body burdens in the exposed animals
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Elfer, Katherine N; Sholl, Andrew B; Wang, Mei; Tulman, David B; Mandava, Sree H; Lee, Benjamin R; Brown, J Quincy
2016-01-01
Real-time on-site histopathology review of biopsy tissues at the point-of-procedure has great potential for significant clinical value and improved patient care. For instance, on-site review can aid in rapid screening of diagnostic biopsies to reduce false-negative results, or in quantitative assessment of biospecimen quality to increase the efficacy of downstream laboratory and histopathology analysis. However, the only currently available rapid pathology method, frozen section analysis (FSA), is too time- and labor-intensive for use in screening large quantities of biopsy tissues and is too destructive for maximum tissue conservation in multiple small needle core biopsies. In this work we demonstrate the spectrally-compatible combination of the nuclear stain DRAQ5 and the anionic counterstain eosin as a dual-component fluorescent staining analog to hematoxylin and eosin intended for use on fresh, unsectioned tissues. Combined with optical sectioning fluorescence microscopy and pseudo-coloring algorithms, DRAQ5 and eosin ("D&E") enables very fast, non-destructive psuedohistological imaging of tissues at the point-of-acquisition with minimal tissue handling and processing. D&E was validated against H&E on a one-to-one basis on formalin-fixed paraffin-embedded and frozen section tissues of various human organs using standard epi-fluorescence microscopy, demonstrating high fidelity of the staining mechanism as an H&E analog. The method was then applied to fresh, whole 18G renal needle core biopsies and large needle core prostate biospecimen biopsies using fluorescence structured illumination optical sectioning microscopy. We demonstrate the ability to obtain high-resolution histology-like images of unsectioned, fresh tissues similar to subsequent H&E staining of the tissue. The application of D&E does not interfere with subsequent standard-of-care H&E staining and imaging, preserving the integrity of the tissue for thorough downstream analysis. These results indicate
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Directory of Open Access Journals (Sweden)
Katherine N Elfer
Full Text Available Real-time on-site histopathology review of biopsy tissues at the point-of-procedure has great potential for significant clinical value and improved patient care. For instance, on-site review can aid in rapid screening of diagnostic biopsies to reduce false-negative results, or in quantitative assessment of biospecimen quality to increase the efficacy of downstream laboratory and histopathology analysis. However, the only currently available rapid pathology method, frozen section analysis (FSA, is too time- and labor-intensive for use in screening large quantities of biopsy tissues and is too destructive for maximum tissue conservation in multiple small needle core biopsies. In this work we demonstrate the spectrally-compatible combination of the nuclear stain DRAQ5 and the anionic counterstain eosin as a dual-component fluorescent staining analog to hematoxylin and eosin intended for use on fresh, unsectioned tissues. Combined with optical sectioning fluorescence microscopy and pseudo-coloring algorithms, DRAQ5 and eosin ("D&E" enables very fast, non-destructive psuedohistological imaging of tissues at the point-of-acquisition with minimal tissue handling and processing. D&E was validated against H&E on a one-to-one basis on formalin-fixed paraffin-embedded and frozen section tissues of various human organs using standard epi-fluorescence microscopy, demonstrating high fidelity of the staining mechanism as an H&E analog. The method was then applied to fresh, whole 18G renal needle core biopsies and large needle core prostate biospecimen biopsies using fluorescence structured illumination optical sectioning microscopy. We demonstrate the ability to obtain high-resolution histology-like images of unsectioned, fresh tissues similar to subsequent H&E staining of the tissue. The application of D&E does not interfere with subsequent standard-of-care H&E staining and imaging, preserving the integrity of the tissue for thorough downstream analysis
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Wang, Guowei; Zhao, Fang; Gao, Ying
2014-12-01
A novel post-chemiluminescence (PCL) reaction was discovered when lercanidipine was injected into the CL reaction mixture of N-chlorosuccinimide with alkaline eosin Y in the presence of cetyltrimethylammonium bromide (CTAB), where eosin Y was used as the CL reagent and CTAB as the surfactant. Based on this observation, a simple and highly sensitive PCL method combined with a flow injection (FI) technique was developed for the assay of lercanidipine. Under optimum conditions, the CL signal was linearly related to the concentration of lercanidipine in the range 7.0 × 10(-10) to 3.0 × 10(-6) g/mL with a detection limit of 2.3 × 10(-10) g/mL (3σ). The relative standard deviation (RSD) was 2.1% for 1.0 × 10(-8) g/mL lercanidipine (n = 13). The proposed method had been applied to the estimation of lercanidipine in tablets and human serum samples with satisfactory results. The possible CL mechanism is also discussed briefly. Copyright © 2014 John Wiley & Sons, Ltd.
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Cytometric analysis of irradiation damaged chromosomes
International Nuclear Information System (INIS)
Wilder, M.E.; Raju, M.R.
1982-01-01
Irradiation of cells in interphase results in dose-dependent damage to DNA which is discernable by flow-cytometric analysis of chromosomes. The quantity (and possibly the quality) of chromosomal changes is different in survival-matched doses of x and α irradiation. It may, therefore, be possible to use these methods for analysis of dose and type of exposure in unknown cases
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Wong, Linda; Hill, Beth L; Hunsberger, Benjamin C; Bagwell, C Bruce; Curtis, Adam D; Davis, Bruce H
2015-01-01
Leuko64™ (Trillium Diagnostics) is a flow cytometric assay that measures neutrophil CD64 expression and serves as an in vitro indicator of infection/sepsis or the presence of a systemic acute inflammatory response. Leuko64 assay currently utilizes QuantiCALC, a semiautomated software that employs cluster algorithms to define cell populations. The software reduces subjective gating decisions, resulting in interanalyst variability of state modeling (PSM). Four hundred and fifty-seven human blood samples were processed using the Leuko64 assay. Samples were analyzed on four different flow cytometer models: BD FACSCanto II, BD FACScan, BC Gallios/Navios, and BC FC500. A probability state model was designed to identify calibration beads and three leukocyte subpopulations based on differences in intensity levels of several parameters. PSM automatically calculates CD64 index values for each cell population using equations programmed into the model. GemStone software uses PSM that requires no operator intervention, thus totally automating data analysis and internal quality control flagging. Expert analysis with the predicate method (QuantiCALC) was performed. Interanalyst precision was evaluated for both methods of data analysis. PSM with GemStone correlates well with the expert manual analysis, r(2) = 0.99675 for the neutrophil CD64 index values with no intermethod bias detected. The average interanalyst imprecision for the QuantiCALC method was 1.06% (range 0.00-7.94%), which was reduced to 0.00% with the GemStone PSM. The operator-to-operator agreement in GemStone was a perfect correlation, r(2) = 1.000. Automated quantification of CD64 index values produced results that strongly correlate with expert analysis using a standard gate-based data analysis method. PSM successfully evaluated flow cytometric data generated by multiple instruments across multiple lots of the Leuko64 kit in all 457 cases. The probability-based method provides greater objectivity, higher
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Directory of Open Access Journals (Sweden)
Pauline Wimberger
2002-01-01
Full Text Available In gynecologic oncology valid prognostic factors are necessary to estimate the course of disease and to define biologically similar subgroups for analysis of therapeutic efficacy. The presented study is a prospective study concerning prognostic significance of DNA ploidy and S‐phase fraction in breast cancer following enrichment of tumor cells by cytokeratin labelling. Epithelial cells were labeled by FITC‐conjugated cytokeratin antibody (CK 5, 6, 8, and CK 17 prior to flow cytometric cell cycle analysis in 327 fresh specimens of primary breast cancer. Univariate analysis in breast cancer detected the prognostic significance of DNA‐ploidy, S‐phase fraction and CV (coefficient of variation of G0G1‐peak of tumor cells for clinical outcome, especially for nodal‐negative patients. Multivariate analysis could not confirm prognostic evidence of DNA‐ploidy and S‐phase fraction. In conclusion, in breast cancer no clinical significance for determination of DNA‐parameters was found.
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Mohamed Gamal Mohamed
2017-03-01
Full Text Available In this study, we synthesized an unusual 2-aminobutyl maleimide isobutyl polyhedral oligomeric silsesquioxane (MIPOSS-NHBu monomer lacking conventional fluorescent groups. We then prepared poly(styrene-alt-2-aminobutyl maleimide isobutyl POSS [poly(S-alt-MIPOSS-NHBu] and poly(4-acetoxystyrene-alt-2-aminobutyl maleimide isobutyl POSS [poly(AS-alt-MIPOSS-NHBu] copolymers through facile free radical copolymerizations using azobisisobutyronitrile as the initiator and tetrahydrofuran as the solvent. A poly(4-hydroxystyrene-alt-2-aminobutyl maleimide isobutyl POSS [poly(HS-alt-MIPOSS-NHBu] copolymer was prepared through acetoxyl hydrazinolysis of poly(AS-alt-MIPOSS-NHBu. We employed 1H, 13C, and 29Si nuclear magnetic resonance spectroscopy; Fourier transform infrared spectroscopy; differential scanning calorimetry; and photoluminescence spectroscopy to investigate the structures and the thermal and optical properties of the monomers and novel POSS-containing alternating copolymers. Intramolecular hydrogen bonding between the amino and dihydrofuran-2,5-dione group and clustering of the locked C=O groups from the POSS nanoparticles in the MIPOSS-NHBu units restricted the intramolecular motion of the polymer chain, causing it to exhibit strong light emission. As a result, the MIPOSS-NHBu monomer and the poly(AS-alt-MIPOSS-NHBu copolymer both have potential applicability in the detection of metal ions with good selectivity.
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Manger, Ronald; Woodle, Doug; Berger, Andrew; Dickey, Robert W; Jester, Edward; Yasumoto, Takeshi; Lewis, Richard; Hawryluk, Timothy; Hungerford, James
2014-01-01
Ciguatoxins are potent neurotoxins with a significant public health impact. Cytotoxicity assays have allowed the most sensitive means of detection of ciguatoxin-like activity without reliance on mouse bioassays and have been invaluable in studying outbreaks. An improvement of these cell-based assays is presented here in which rapid flow cytometric detection of ciguatoxins and saxitoxins is demonstrated using fluorescent voltage sensitive dyes. A depolarization response can be detected directly due to ciguatoxin alone; however, an approximate 1000-fold increase in sensitivity is observed in the presence of veratridine. These results demonstrate that flow cytometric assessment of ciguatoxins is possible at levels approaching the trace detection limits of our earlier cytotoxicity assays, however, with a significant reduction in analysis time. Preliminary results are also presented for detection of brevetoxins and for automation and throughput improvements to a previously described method for detecting saxitoxins in shellfish extracts.
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Flow cytometric probing of mitochondrial function in equine peripheral blood mononuclear cells
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Coignoul Freddy
2007-09-01
Full Text Available Abstract Background The morphopathological picture of a subset of equine myopathies is compatible with a primary mitochondrial disease, but functional confirmation in vivo is still pending. The cationic dye JC-1 exhibits potential-dependent accumulation in mitochondria that is detectable by a fluorescence shift from green to orange. As a consequence, mitochondrial membrane potential can be optically measured by the orange/green fluorescence intensity ratio. A flow cytometric standardized analytic procedure of the mitochondrial function of equine peripheral blood mononuclear cells is proposed along with a critical appraisal of the crucial questions of technical aspects, reproducibility, effect of time elapsed between blood sampling and laboratory processing and reference values. Results The JC-1-associated fluorescence orange and green values and their ratio were proved to be stable over time, independent of age and sex and hypersensitive to intoxication with a mitochondrial potential dissipator. Unless time elapsed between blood sampling and laboratory processing does not exceed 5 hours, the values retrieved remain stable. Reference values for clinically normal horses are given. Conclusion Whenever a quantitative measurement of mitochondrial function in a horse is desired, blood samples should be taken in sodium citrate tubes and kept at room temperature for a maximum of 5 hours before the laboratory procedure detailed here is started. The hope is that this new test may help in confirming, studying and preventing equine myopathies that are currently imputed to mitochondrial dysfunction.
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Gauthier, J.D.; Jenkins, J.A.; La Peyre, Jerome F.
2004-01-01
Parasite surface glycoconjugates are frequently involved in cellular recognition and colonization of the host. This study reports on the identification of Perkinsus marinus surface carbohydrates by flow cytometric analyses of fluorescein isothiocyanate-conjugated lectin binding. Lectin-binding specificity was confirmed by sugar inhibition and Kolmogorov-Smirnov statistics. Clear, measurable fluorescence peaks were discriminated, and no parasite autofluorescence was observed. Parasites (GTLA-5 and Perkinsus-1 strains) harvested during log and stationary phases of growth in a protein-free medium reacted strongly with concanavalin A and wheat germ agglutinin, which bind to glucose-mannose and N-acetyl-D-glucosamine (GlcNAc) moieties, respectively. Both P. marinus strains bound with lower intensity to Maclura pomifera agglutinin, Bauhinia purpurea agglutinin, soybean agglutinin (N-acetyl-D-galactosamine-specific lectins), peanut agglutinin (PNA) (terminal galactose specific), and Griffonia simplicifolia II (GlcNAc specific). Only background fluorescence levels were detected with Ulex europaeus agglutinin I (L-fucose specific) and Limulus polyphemus agglutinin (sialic acid specific). The lectin-binding profiles were similar for the 2 strains except for a greater relative binding intensity of PNA for Perkinsus-1 and an overall greater lectin-binding capacity of Perkinsus-1 compared with GTLA-5. Growth stage comparisons revealed increased lectin-binding intensities during stationary phase compared with log phase of growth. This is the first report of the identification of surface glycoconjugates on a Perkinsus spp. by flow cytometry and the first to demonstrate that differential surface sugar expression is growth phase and strain dependent. ?? American Society of Parasitologists 2004.
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García, Francisca C.
2015-07-17
The ’cytometric diversity’ of phytoplankton communities has been studied based on single-cell properties, but the applicability of this method to characterize bacterioplankton has been unexplored. Here, we analysed seasonal changes in cytometric diversity of marine bacterioplankton along a decadal time-series at three coastal stations in the Southern Bay of Biscay. Shannon-Weaver diversity estimates and Bray-Curtis similarities obtained by cytometric and molecular (16S rRNA tag sequencing) methods were significantly correlated in samples from a 3.5-year monthly time-series. Both methods showed a consistent cyclical pattern in the diversity of surface bacterial communities with maximal values in winter. The analysis of the highly resolved flow cytometry time-series across the vertical profile showed that water column mixing was a key factor explaining the seasonal changes in bacterial composition and the winter increase in bacterial diversity in coastal surface waters. Due to its low cost and short processing time as compared to genetic methods, the cytometric diversity approach represents a useful complementary tool in the macroecology of aquatic microbes.
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VRIELING, EG; DRAAIJER, A; VANZEIJL, WJM; PEPERZAK, L; GIESKES, WWC; VEENHUIS, M; Zeijl, Wilhelmus J.M. van
Two different fluorescein isothiocyanate (FITC) conjugates were used to analyze the effect of labeling intensity on the flow cytometric appearance of marine dinoflagellates labeled with antibodies that specifically recognized the outer cell wall. Location of the labeling was revealed by
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Zhai, Juping; Ding, Mengyuan; Yang, Tianjie; Zuo, Bin; Weng, Zhen; Zhao, Yunxiao; He, Jun; Wu, Qingyu; Ruan, Changgeng; He, Yang
2017-10-23
Platelet autoantibody detection is critical for immune thrombocytopenia (ITP) diagnosis and prognosis. Therefore, we aimed to establish a quantitative flow cytometric immunobead assay (FCIA) for ITP platelet autoantibodies evaluation. Capture microbeads coupled with anti-GPIX, -GPIb, -GPIIb, -GPIIIa and P-selectin antibodies were used to bind the platelet-bound autoantibodies complex generated from plasma samples of 250 ITP patients, 163 non-ITP patients and 243 healthy controls, a fluorescein isothiocyanate (FITC)-conjugated secondary antibody was the detector reagent and mean fluorescence intensity (MFI) signals were recorded by flow cytometry. Intra- and inter-assay variations of the quantitative FCIA assay were assessed. Comparisons of the specificity, sensitivity and accuracy between quantitative and qualitative FCIA or monoclonal antibody immobilization of platelet antigen (MAIPA) assay were performed. Finally, treatment process was monitored by our quantitative FCIA in 8 newly diagnosed ITPs. The coefficient of variations (CV) of the quantitative FCIA assay were respectively 9.4, 3.8, 5.4, 5.1 and 5.8% for anti-GPIX, -GPIb, -GPIIIa, -GPIIb and -P-selectin autoantibodies. Elevated levels of autoantibodies against platelet glycoproteins GPIX, GPIb, GPIIIa, GPIIb and P-selectin were detected by our quantitative FCIA in ITP patients compared to non-ITP patients or healthy controls. The sensitivity, specificity and accuracy of our quantitative assay were respectively 73.13, 81.98 and 78.65% when combining all 5 autoantibodies, while the sensitivity, specificity and accuracy of MAIPA assay were respectively 41.46, 90.41 and 72.81%. A quantitative FCIA assay was established. Reduced levels of platelet autoantibodies could be confirmed by our quantitative FCIA in ITP patients after corticosteroid treatment. Our quantitative assay is not only good for ITP diagnosis but also for ITP treatment monitoring.
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Marinic, Karlo; Manoil, Daniel; Filieri, Anna; Wataha, John C; Schrenzel, Jacques; Lange, Norbert; Bouillaguet, Serge
2015-09-01
In dentistry, antibacterial photodynamic therapy (a-PDT) has shown promising results for inactivating bacterial biofilms causing carious, endodontic and periodontal diseases. In the current study, we assessed the ability of eosin Y exposed to 3 irradiation protocols at inactivating Enterococcus faecalis biofilms, in vitro. E. faecalis biofilms formed on hydroxyapatite disks were incubated with eosin Y (10-80μM), then activated with blue light using different irradiation protocols. Biofilms exposed to continuous exposure were incubated for 40min before being light-activated for 960 s. For the intermittent exposure, biofilms were exposed 4 times to the light/photosensitizer combination (960 s total) without renewing the photosensitizer. For repeated a-PDT, the same light dose was delivered in a series of 4 irradiation periods separated by dark periods; fresh photosensitizer was added between each light irradiation. After treatment, bacteria were immediately labeled with LIVE/DEAD BacLight Bacterial Viability kit and viability was assessed by flow cytometry (FCM). Results were statistically analyzed using one-way ANOVA and Tukey multiple comparison intervals (α=0.05). The viability of E. faecalis biofilms exposed to 10μM eosin Y, was significantly reduced compared to controls (light only-eosin Y only). After a second exposure to blue light-activated eosin Y, viability significantly decreased from 58% to 12% whereas 6.5% of the bacterial biofilm remained live after a third exposure (p<0.05). Only 3.5% of the bacterial population survived after the fourth exposure. The results of this study indicate that blue light-activated eosin Y can photoinactivate E. faecalis biofilms grown on hydroxyapatite disks. Also, repeated exposures to blue light-activated eosin Y were shown to significantly improve efficacy. Further studies seem warranted to optimize the antibacterial activity of blue light-activated eosin Y on major oral pathogens. Copyright © 2015 Elsevier B.V. All
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Genome size variation among and within Camellia species by using flow cytometric analysis.
Directory of Open Access Journals (Sweden)
Hui Huang
Full Text Available BACKGROUND: The genus Camellia, belonging to the family Theaceae, is economically important group in flowering plants. Frequent interspecific hybridization together with polyploidization has made them become taxonomically "difficult taxa". The DNA content is often used to measure genome size variation and has largely advanced our understanding of plant evolution and genome variation. The goals of this study were to investigate patterns of interspecific and intraspecific variation of DNA contents and further explore genome size evolution in a phylogenetic context of the genus. METHODOLOGY/PRINCIPAL FINDINGS: The DNA amount in the genus was determined by using propidium iodide flow cytometry analysis for a total of 139 individual plants representing almost all sections of the two subgenera, Camellia and Thea. An improved WPB buffer was proven to be suitable for the Camellia species, which was able to counteract the negative effects of secondary metabolite and generated high-quality results with low coefficient of variation values (CV <5%. Our results showed trivial effects on different tissues of flowers, leaves and buds as well as cytosolic compounds on the estimation of DNA amount. The DNA content of C. sinensis var. assamica was estimated to be 1C = 3.01 pg by flow cytometric analysis, which is equal to a genome size of about 2940 Mb. CONCLUSION: Intraspecific and interspecific variations were observed in the genus Camellia, and as expected, the latter was larger than the former. Our study suggests a directional trend of increasing genome size in the genus Camellia probably owing to the frequent polyploidization events.
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Hadjichristidis, Nikolaos
2015-05-26
A novel strategy towards well-defined polymethylene-based co/terpolymers, by combining anthracene/maleimide Diels-Alder reaction with polyhomologation, is presented. For the synthesis of diblock copolymers the following approach was applied: a) synthesis of α-anthracene-ω-hydroxy- polymethylene by polyhomologation using tri (9-anthracene-methyl propyl ether) borane as initiator, b) synthesis of furan-protected-maleimide-terminated poly (ε-caprolactone) or polyethylene glycol and c). Diels-Alder reaction between the anthracene and maleimide-terminated polymers. In the case of triblock terpolymers the α-anthracene-ω-hydroxy-polymethylene was used as macroinitiator for the ring-opening polymerization of D, L-lactide to afford an anthracene-terminated PM-b-PLA copolymer, followed by Diels-Alder reaction with furan-protected maleimide-terminated poly (ε-caprolactone) or polyethylene glycol to give the triblock terpolymers. All intermediate and final products were characterized by SEC, 1H NMR, UV-VIS spectroscopy and DSC.
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Flow cytometric kinetic assay of the activity of Na+/H+ antiporter in mammalian cells.
Dolz, María; O'Connor, José-Enrique; Lequerica, Juan L
2004-10-01
The Na(+)/H(+) exchanger (NHE) of mammalian cells is an integral membrane protein that extrudes H(+) ion in exchange for extracellular Na(+) and plays a crucial role in the regulation of intracellular pH (pHi). Thus, when pHi is lowered, NHE extrudes protons at a rate depending of pHi that can be expressed as pH units/s. To abolish the activity of other cellular pH-restoring systems, cells were incubated in bicarbonate-free Dulbecco's modified Eagle's medium buffered with HEPES. Flow cytometry was used to determine pHi with 2',7'-bis-(2-carboxyethyl)-5-(and-6)-carboxyfluorescein acetoxymethyl ester or 5-(and-6)-carboxy SNARF-1 acetoxymethyl ester acetate, and the appropriate fluorescence ratios were measured. The calibration of fluorescence ratios versus pHi was established by using ionophore nigericin. The activity of NHE was calculated by a kinetic flow cytometric assay as the slope at time 0 of the best-fit curve of pHi recovery versus time after intracellular acidification with a pulse of exogenous sodium propionate. The kinetic method allowed determination of the pHi-dependent activity of NHE in cell lines and primary cell cultures. NHE activity values were demonstrated to be up to 0.016 pH units/s within the pHi range of 7.3 to 6.3. The inhibition of NHE activity by the specific inhibitor ethyl isopropyl amiloride was easily detected by this method. The assay conditions can be used to relate variations in pHi with the activity of NHE and provide a standardized method to compare between different cells, inhibitors, models of ischemia by acidification, and other relevant experimental or clinical situations.
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Flow cytometric evaluation of physico-chemical impact on Gram-positive and Gram-negative bacteria
Fröhling, Antje; Schlüter, Oliver
2015-01-01
Since heat sensitivity of fruits and vegetables limits the application of thermal inactivation processes, new emerging inactivation technologies have to be established to fulfill the requirements of food safety without affecting the produce quality. The efficiency of inactivation treatments has to be ensured and monitored. Monitoring of inactivation effects is commonly performed using traditional cultivation methods which have the disadvantage of the time span needed to obtain results. The aim of this study was to compare the inactivation effects of peracetic acid (PAA), ozonated water (O3), and cold atmospheric pressure plasma (CAPP) on Gram-positive and Gram-negative bacteria using flow cytometric methods. E. coli cells were completely depolarized after treatment (15 s) with 0.25% PAA at 10°C, and after treatment (10 s) with 3.8 mg l−1 O3 at 12°C. The membrane potential of CAPP treated cells remained almost constant at an operating power of 20 W over a time period of 3 min, and subsequently decreased within 30 s of further treatment. Complete membrane permeabilization was observed after 10 s O3 treatment, but treatment with PAA and CAPP did not completely permeabilize the cells within 2 and 4 min, respectively. Similar results were obtained for esterase activity. O3 inactivates cellular esterase but esterase activity was detected after 4 min CAPP treatment and 2 min PAA treatment. L. innocua cells and P. carotovorum cells were also permeabilized instantaneously by O3 treatment at concentrations of 3.8 ± 1 mg l−1. However, higher membrane permeabilization of L. innocua and P. carotovorum than of E. coli was observed at CAPP treatment of 20 W. The degree of bacterial damage due to the inactivation processes is highly dependent on treatment parameters as well as on treated bacteria. Important information regarding the inactivation mechanisms can be obtained by flow cytometric measurements and this enables the definition of critical process parameters. PMID
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Flow cytometric evaluation of physico-chemical impact on Gram-positive and Gram-negative bacteria
Directory of Open Access Journals (Sweden)
Antje eFröhling
2015-09-01
Full Text Available Since heat sensitivity of fruits and vegetables limits the application of thermal inactivation processes, new emerging inactivation technologies have to be established to fulfil the requirements of food safety without affecting the produce quality. The efficiency of inactivation treatments has to be ensured and monitored. Monitoring of inactivation effects is commonly performed using traditional cultivation methods which have the disadvantage of the time span needed to obtain results.The aim of this study was to compare the inactivation effects of peracetic acid (PAA, ozonated water (O3 and cold atmospheric pressure plasma (CAPP on Gram-positive and Gram-negative bacteria using flow cytometric methods. E. coli cells were completely depolarized after treatment (15 s with 0.25 % PAA at 10 °C, and after treatment (10 s with 3.8 mg l-1 O3 at 12°C. The membrane potential of CAPP treated cells remained almost constant at an operating power of 20 W over a time period of 3 min, and subsequently decreased within 30 s of further treatment. Complete membrane permeabilization was observed after 10 s O3 treatment, but treatment with PAA and CAPP did not completely permeabilize the cells within 2 min and 4 min, respectively. Similar results were obtained for esterase activity. O3 inactivates cellular esterase but esterase activity was detected after 4 min CAPP treatment and 2 min PAA treatment. L. innocua cells and P. carotovorum cells were also permeabilized instantaneously by O3 treatment at concentrations of 3.8 ± 1 mg l-1. However, higher membrane permeabilization of L. innocua and P. carotovorum than of E. coli was observed at CAPP treatment of 20 W. The degree of bacterial damage due to the inactivation processes is highly dependent on treatment parameters as well as on treated bacteria. Important information regarding the inactivation mechanisms can be obtained by flow cytometric measurements and this enables the definition of critical process
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Yu, Yen-Rei A.; O’Koren, Emily G.; Hotten, Danielle F.; Kan, Matthew J.; Kopin, David; Nelson, Erik R.; Que, Loretta; Gunn, Michael D.
2016-01-01
Flow cytometry is used extensively to examine immune cells in non-lymphoid tissues. However, a method of flow cytometric analysis that is both comprehensive and widely applicable has not been described. We developed a protocol for the flow cytometric analysis of non-lymphoid tissues, including methods of tissue preparation, a 10-fluorochrome panel for cell staining, and a standardized gating strategy, that allows the simultaneous identification and quantification of all major immune cell types in a variety of normal and inflamed non-lymphoid tissues. We demonstrate that our basic protocol minimizes cell loss, reliably distinguishes macrophages from dendritic cells (DC), and identifies all major granulocytic and mononuclear phagocytic cell types. This protocol is able to accurately quantify 11 distinct immune cell types, including T cells, B cells, NK cells, neutrophils, eosinophils, inflammatory monocytes, resident monocytes, alveolar macrophages, resident/interstitial macrophages, CD11b- DC, and CD11b+ DC, in normal lung, heart, liver, kidney, intestine, skin, eyes, and mammary gland. We also characterized the expression patterns of several commonly used myeloid and macrophage markers. This basic protocol can be expanded to identify additional cell types such as mast cells, basophils, and plasmacytoid DC, or perform detailed phenotyping of specific cell types. In examining models of primary and metastatic mammary tumors, this protocol allowed the identification of several distinct tumor associated macrophage phenotypes, the appearance of which was highly specific to individual tumor cell lines. This protocol provides a valuable tool to examine immune cell repertoires and follow immune responses in a wide variety of tissues and experimental conditions. PMID:26938654
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International Nuclear Information System (INIS)
Welleweerd, J.; Wilder, M.E.; Carpenter, S.G.; Raju, M.R.
1984-01-01
Chinese hamster M3-1 cells were irradiated with several doses of x rays or α particles from 238 Pu. Propidium iodide-stained chromosome suspensions were prepared at different times after irradiation; cells were also assayed for survival. The DNA histograms of these chromosomes showed increased background counts with increased doses of radiation. This increase in background was cell-cycle dependent and was correlated with cell survival. The correlation between radiation-induced chromosome damage and cell survival was the same for X rays and α particles. Data are presented which indicate that flow cytometric analysis of chromosomes of irradiated cell populations can be a useful adjunct to classical cytogenic analysis of irradiation-induced chromosomal damage by virtue of its ability to express and measure chromosomal damage not seen by classical cytogenic methods
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Automatic analysis of flow cytometric DNA histograms from irradiated mouse male germ cells
International Nuclear Information System (INIS)
Lampariello, F.; Mauro, F.; Uccelli, R.; Spano, M.
1989-01-01
An automatic procedure for recovering the DNA content distribution of mouse irradiated testis cells from flow cytometric histograms is presented. First, a suitable mathematical model is developed, to represent the pattern of DNA content and fluorescence distribution in the sample. Then a parameter estimation procedure, based on the maximum likelihood approach, is constructed by means of an optimization technique. This procedure has been applied to a set of DNA histograms relative to different doses of 0.4-MeV neutrons and to different time intervals after irradiation. In each case, a good agreement between the measured histograms and the corresponding fits has been obtained. The results indicate that the proposed method for the quantitative analysis of germ cell DNA histograms can be usefully applied to the study of the cytotoxic and mutagenic action of agents of toxicological interest such as ionizing radiations.18 references
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adsorption, eosin, humic, peat
anshar, andi muhammad
2015-01-01
Eosin is one of the dyes commonly used in the industry and has the potential to cause pollution of the water environment. The Eosin pollution treatment methods used in this study was the adsorption method using humin fraction obtained from the peat land comes from Kalimantan. From the research data showed that the adsorption of eosin in humin result of washing with HCl / HF optimum at pH 4 and a contact time of 60 minutes with the adsorption-order rate was 8,4 x 10-3 min-1
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Multivariate analysis of flow cytometric data using decision trees
Directory of Open Access Journals (Sweden)
Svenja eSimon
2012-04-01
Full Text Available Characterization of the response of the host immune system is important in understanding the bidirectional interactions between the host and microbial pathogens. For research on the host site, flow cytometry has become one of the major tools in immunology. Advances in technology and reagents allow now the simultaneous assessment of multiple markers on a single cell level generating multidimensional data sets that require multivariate statistical analysis. We explored the explanatory power of the supervised machine learning method called 'induction of decision trees' in flow cytometric data. In order to examine whether the production of a certain cytokine is depended on other cytokines, datasets from intracellular staining for six cytokines with complex patterns of co-expression were analyzed by induction of decision trees. After weighting the data according to their class probabilities, we created a total of 13,392 different decision trees for each given cytokine with different parameter settings. For a more realistic estimation of the decision trees's quality, we used stratified 5-fold cross-validation and chose the 'best' tree according to a combination of different quality criteria. While some of the decision trees reflected previously known co-expression patterns, we found that the expression of some cytokines was not only dependent on the co-expression of others per se, but was also dependent on the intensity of expression. Thus, for the first time we successfully used induction of decision trees for the analysis of high dimensional flow cytometric data and demonstrated the feasibility of this method to reveal structural patterns in such data sets.
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The unexpected product of Diels-Alder reaction between "indanocyclon" and maleimide
Dobrowolski, Michał A.; Roszkowski, Piotr; Struga, Marta; Szulczyk, Daniel
2017-02-01
A heterocyclic compound commonly known as "indanocyclon" undergoes an unexpected Diels-Alder addition with maleimide. The resulting product has been isolated and characterized in order to get an information about its structure and possible mechanism of the reaction. Extensive comparison of single crystal properties of 3-(2,8-dioxo-1,3-diphenyl-2,8-dihydrocyclopenta[a]inden-8a(1H)-yl)pyrrolidine-2,5-dione and favorable product of the reaction has been also performed.
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Keserue, Hans-Anton; Füchslin, Hans Peter; Egli, Thomas
2011-01-01
Giardia lamblia is an important waterborne pathogen and is among the most common intestinal parasites of humans worldwide. Its fecal-oral transmission leads to the presence of cysts of this pathogen in the environment, and so far, quantitative rapid screening methods are not available for various matrices, such as surface waters, wastewater, or food. Thus, it is necessary to establish methods that enable reliable rapid detection of a single cyst in 10 to 100 liters of drinking water. Conventional detection relies on cyst concentration, isolation, and confirmation by immunofluorescence microscopy (IFM), resulting in low recoveries and high detection limits. Many different immunomagnetic separation (IMS) procedures have been developed for separation and cyst purification, so far with variable but high losses of cysts. A method was developed that requires less than 100 min and consists of filtration, resuspension, IMS, and flow cytometric (FCM) detection. MACS MicroBeads were used for IMS, and a reliable flow cytometric detection approach was established employing 3 different parameters for discrimination from background signals, i.e., green and red fluorescence (resulting from the distinct pattern emitted by the fluorescein dye) and sideward scatter for size discrimination. With spiked samples, recoveries exceeding 90% were obtained, and false-positive results were never encountered for negative samples. Additionally, the method was applicable to naturally occurring cysts in wastewater and has the potential to be automated. PMID:21685159
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International Nuclear Information System (INIS)
Ludwikow, G.; Staalnacke, C.G.; Johanson, K.J.; Sundell-Bergman, S.; Richter, S.; Swedish Univ. of Agricultural Sciences, Uppsala; Uppsala Univ.
1990-01-01
Iododeoxyuridine labelled (IUdR(+)) and unlabelled (IUdR(-)) CHO cells irradiated with 2 Gy of soft X-rays showed only minor differences in the kinetics of micronuclei formation during the first 20 hours postirradiation period. Between 20 to 40 hours, the IUdR(-) cells showed approximately a constant number of micronuclei while the number of micronuclei in IUdR(+) cells was still increasing. The frequency of micronuclei was higher in IUdR(+) cells compared to IUdR(-) cells at 24 hours after irradiation with various doses up to 4.0 Gy. Dose modifying factors were found to be 1.3 (microscopic evaluation) and 1.8 (flow cytometric evaluation). Flow cytometry with use of two parameters, fluorescence from propidium iodide and light scattering, seems to be a good tool to estimate the frequency of micronuclei in CHO cells in the dose range up to about 4 Gy. At higher doses perturbation of the cell cycle and the appearance of dying cells will influence the results. (orig.)
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Guy, Julia; Caron, Karine; Dufresne, Stéphane; Michnick, Stephen W; Skene, W G; Keillor, Jeffrey W
2007-10-03
Dimaleimide fluorogens are being developed for application to fluorescent protein labeling. In this method, fluorophores bearing two maleimide quenching groups do not fluoresce until both maleimide groups have undergone thiol addition reactions with the Cys residues of the target protein sequence [J. Am. Chem. Soc. 2005, 127, 559-566]. In this work, a new convergent synthetic route was developed that would allow any fluorophore to be attached via a linker to a dimaleimide moiety in a modular fashion. Series of dimaleimide and dansyl derivatives were thus prepared conveniently and used to elucidate the mechanism of maleimide quenching. Intersystem crossing was ruled out as a potential quenching pathway, based on the absence of a detectable triplet intermediate by laser flash photolysis. Stern-Volmer rate constants were measured with exogenous dimaleimide quenchers and found to be close to the diffusion-controlled limits, consistent with electron transfer being thermodynamically favorable. The thermodynamic feasibility of the photoinduced electron transfer (PET) quenching mechanism was verified by cyclic voltammetry. The redox potentials measured for dansyl and maleimide confirm that electron transfer from the dansyl excited state to a pendant maleimide group is exergonic and is responsible for fluorescence quenching of the fluorogens studied herein. Taking this PET quenching mechanism into account, future fluorogenic protein labeling agents will be designed with spacers of variable length and rigidity to probe the structure-property PET efficiency relationship.
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Kapasitas dan Energi Adsorpsi Humin Terhadap Eosin
Directory of Open Access Journals (Sweden)
Anshar A.M.
2015-07-01
Full Text Available Humin is a natural ingredient that is widely available in Indonesia, especially Borneo. Humin is interact and absorb eosin compound that can cause environmental pollution. The interaction between eosin and humin was set on optimum time dan pH so that the humin adsorption would be optimum. This research measured the adsorption capacity and energy required to adsorp eosin on humin by performing various concentration of eosin. The amount of humin adsorption energy against eosin was 34.136 kJ/mol while the adsorption capacity was1, 611 x 10-6mol/g. This indicated that the type of interaction between humin and eosin was chemical interaction.
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Kapasitas dan Energi Adsorpsi Humin Terhadap Eosin
Anshar A.M.; Santosa S. J.; Sudiono S.
2015-01-01
Humin is a natural ingredient that is widely available in Indonesia, especially Borneo. Humin is interact and absorb eosin compound that can cause environmental pollution. The interaction between eosin and humin was set on optimum time dan pH so that the humin adsorption would be optimum. This research measured the adsorption capacity and energy required to adsorp eosin on humin by performing various concentration of eosin. The amount of humin adsorption energy against eosin was 34.136 kJ/mo...
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Energy Technology Data Exchange (ETDEWEB)
Nakagawa, Seiko, E-mail: Nakagawa.Seiko@iri-tokyo.jp [Tokyo Metropolitan Industrial Technology Research Institute, 2-4-10 Aoumi, Koto-ku, Tokyo 135-0064 (Japan); Taguchi, Mitsumasa; Kimura, Atsushi [Environmental Radiation Processing Group, Environment and Industrial Materials Research Division, Quantum Beam Science Directorate, Japan Atomic Energy Agency, 1233 Watanuki, Takasaki, Gunma 370-1292 (Japan)
2011-11-15
N{sub 2}-saturated 2-propanol solutions containing styrene and maleimide were irradiated by electron, proton, He and C-ion beams. The styrene-maleimide copolymer was produced by radical polymerization induced during irradiation. The effects of the radical distribution, which depends on the LET or dose rate, on the molecular weight and the polymer yield were discussed. - Highlights: > Maleimide with styrene in N{sub 2}-saturated 2-propanol was irradiated by electron and heavy ions. > LET and dose rate effects of radical polymerization induced by irradiation were studied. > Results have shown the relation between radical distribution and efficiency of polymerization.
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Ultrafast energy transfer in dansylated POPAM--eosin complexes
Aumanen, Jukka; Lehtovuori, Viivi; Werner, Nicole; Richardt, Gabriele; van Heyst, Jeroen; Vögtle, Fritz; Korppi-Tommola, Jouko
2006-12-01
Excitation energy transfer (EET) in dendritic host-guest complexes has been studied. Three generations G2, G3 and G4 of dansyl substituted poly(propyleneamine) dendrimers (POPAM) were complexed with a fluorescent dye eosin in chloroform solution. Arrival of excitation from dansyls to eosin was monitored by femtosecond transient absorption spectroscopy. EET rates from the dansyls to eosin(s) are characterised by two time constants 1 ps and 6 ps independent of dendrimer generation. Relaxation processes in eosin were clearly faster when complexed with dendrimer than in solution. As several eosins are bound to G3 and G4 dendrimers, besides host-guest interaction, also eosin-eosin interactions may contribute to the faster relaxation observed in these complexes.
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DEFF Research Database (Denmark)
Engelholm, S A; Spang-Thomsen, M; Vindeløv, L L
1986-01-01
A method based on detection of drug-induced cell cycle perturbation by flow cytometric DNA analysis has previously been described in Ehrlich ascites tumors as a way to estimate chemosensitivity. The method is extended to test human small-cell carcinoma of the lung. Three tumors with different...... sensitivities to melphalan in nude mice were used. Tumors were disaggregated by a combined mechanical and enzymatic method and thereafter have incubated with different doses of melphalan. After incubation the cells were plated in vitro on agar, and drug induced cell cycle changes were monitored by flow...
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H-Point Standard Addition Method for Simultaneous Determination of Eosin and Erytrosine
Directory of Open Access Journals (Sweden)
Amandeep Kaur
2011-01-01
Full Text Available A new, simple, sensitive and selective H-point standard addition method (HPSAM has been developed for resolving binary mixture of food colorants eosin and erythrosine, which show overlapped spectra. The method is based on the complexation of food dyes eosin and erythrosine with Fe(III complexing reagent at pH 5.5 and solubilizing complexes in triton x-100 micellar media. Absorbances at the two pairs of wavelengths, 540 and 550 nm (when eosin acts as analyte or 518 and 542 nm (when erythrosine act as analyte were monitored. This method has satisfactorily been applied for the determination of eosin and erythrosine dyes in synthetic mixtures and commercial products.
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International Nuclear Information System (INIS)
Jensen, R.H.; Bigbee, W.L.; Langlois, R.G.; Grant, S.G.; Pleshanov, P.G.; Chirkov, A.A.; Pilinskaya, M.A.
1990-01-01
An analytical technique has been developed that allows laser-based flow cytometric measurement of the frequency of red blood cells that have lost allele-specific expression of a cell surface antigen due to genetic toxicity in bone marrow precursor cells. Previous studies demonstrated a correlation of such effects with the exposure of each individual to mutagenic phenomena, such as ionizing radiation, and the effects can persist for the lifetime of each individual. During the emergency response to the nuclear power plant accident at Chernobyl, Ukraine, USSR, a number of people were exposed to whole body doses of ionizing radiation. Some of these individuals were tested with this laser-based assay and found to express a dose-dependent increase in the frequency of variant red blood cells that appears to be a persistent biological effect. All data indicate that this assay might well be used as a biodosimeter to estimate radiation dose and also as an element to be used for estimating the risk of each individual to develop cancer due to radiation exposure. 17 refs., 5 figs
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Flow cytometric detection of micronuclei by combined staining of DNA and membranes
International Nuclear Information System (INIS)
Wessels, J.M.; Nuesse, M.
1995-01-01
A new staining method is presented for flow cytometric measurement of micronuclei (MN) in cell cultures and human lymphocytes using membrane-specific fluorescent dyes in addition to DNA staining. Several combinations of fluorescent membrane and DNA dyes were studied for a better discrimination of MN from debris in a suspension of nuclei and micronuclei. For staining of membranes, the lipophilic dyes 2-hydroxyethyl-7,12,17-tris(methoxyethyl)porphycene (HEPn) and 1,6-diphenyl-1,3,5-hexatriene (DPH) were used in combination with ethidium bromide (EB), proflavine (PF), and Hoechst 33258 (HO). Due to their spectral properties, HO or EB combined with HEPn were not as suitable for the discrimination of MN from debris as was HEPn in combination with PF. With HEPn in combination with PF, however, additional noise was found at low fluorescence intensities, probably due to free fluorescent dye molecules in the solution. The optimal simultaneous staining of membranes and DNA was obtained using a combination of DPH and EB. The induction of MN in Chinese hamster and mouse NIH-3T3 cells by UV-B illumination was studied with this new staining technique. UV-B illumination (280-360 nm) induced MN in both cell lines. Chinese hamster cells were found to be more sensitive to these wavelengths. Illumination with wavelengths above 360 nm did not induce MN in either cell line. The results obtained from human lymphocytes using the combination of EB or DPH were comparable to the results obtained with the combination of EB and HO. 23 refs., 7 figs
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International Nuclear Information System (INIS)
Schultz, N.; Norrgren, L.; Grawe, J.; Johannisson, A.; Medhage, O.
1993-01-01
Rainbow trout (oncorhynchus mykiss) were exposed to a single X-ray dose of 4 Gy. The frequency of micronuclei in the peripheral erythrocytes was investigated at regular intervals up to 58 days after the exposure. A flow cytometric method and a semi-automatic image analysis method were used to estimate the micronuclei frequency. The results show that both methods can detect an increased frequency of micronuclei in peripheral erythrocytes from exposed fish. However, the semi-automatic image analysis method was the most stable and sensitive. (Author)
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Schelté, P; Boeckler, C; Frisch, B; Schuber, F
2000-01-01
The comparative reactivity of maleimide and bromoacetyl groups with thiols (2-mercaptoethanol, free cysteine, and cysteine residues present at the N-terminus of peptides) was investigated in aqueous media. These studies were performed (i) with water-soluble functionalized model molecules, i.e., polyoxyethylene-based spacer arms that could also be coupled to lipophilic anchors destined to be incorporated into liposomes, and (ii) with small unilamellar liposomes carrying at their surface these thiol-reactive functions. Our results indicate that an important kinetic discrimination (2-3 orders of magnitude in terms of rate constants) can be achieved between the maleimide and bromoacetyl functions when the reactions with thiols are performed at pH 6.5. The bromoacetyl function which reacts at higher pH values (e.g., pH 9.0) retained a high chemoselectivity; i.e., under conditions where it reacted appreciably with the thiols of, e.g., HS-peptides, it did react with other nucleophilic functions such as alpha- and epsilon-amino groups or imidazole, which could also be present in peptides. This differential reactivity was applied to design chemically defined and highly immunogenic liposomal diepitope constructs as synthetic vaccines, i.e., vesicles carrying at their surface two different peptides conjugated each to a specific amphiphilic anchor. This was realized by coupling sequentially at pH 6.5 and 9.0 two HS-peptides to preformed vesicles containing lipophilic anchors functionalized with maleimide and bromoacetyl groups [Boeckler, C., et al. (1999) Eur. J. Immunol. 29, 2297-2308].
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Kawai, Y; Nagai, Y; Ogawa, E; Kondo, H
2017-04-01
To provide target values for the manufacturers' survey of the Japanese Society for Laboratory Hematology (JSLH), accurate standard data from healthy volunteers were needed for the five-part differential leukocyte count. To obtain such data, JSLH required an antibody panel that achieved high specificity (particularly for mononuclear cells) using simple gating procedures. We developed a flow cytometric method for determining the differential leukocyte count (JSLH-Diff) and validated it by comparison with the flow cytometric differential leukocyte count of the International Council for Standardization in Haematology (ICSH-Diff) and the manual differential count obtained by microscopy (Manual-Diff). First, the reference laboratory performed an imprecision study of JSLH-Diff and ICSH-Diff, as well as performing comparison among JSLH-Diff, Manual-Diff, and ICSH-Diff. Then two reference laboratories and seven participating laboratories performed imprecision and accuracy studies of JSLH-Diff, Manual-Diff, and ICSH-Diff. Simultaneously, six manufacturers' laboratories provided their own representative values by using automated hematology analyzers. The precision of both JSLH-Diff and ICSH-Diff methods was adequate. Comparison by the reference laboratory showed that all correlation coefficients, slopes and intercepts obtained by the JSLH-Diff, ICSH-Diff, and Manual-Diff methods conformed to the criteria. When the imprecision and accuracy of JSLH-Diff were assessed at seven laboratories, the CV% for lymphocytes, neutrophils, monocytes, eosinophils, and basophils was 0.5~0.9%, 0.3~0.7%, 1.7~2.6%, 3.0~7.9%, and 3.8~10.4%, respectively. More than 99% of CD45 positive leukocytes were identified as normal leukocytes by JSLH-Diff. When JSLH-Diff method were validated by comparison with Manual-Diff and ICSH-Diff, JSLH-Diff showed good performance as a reference method. © 2016 John Wiley & Sons Ltd.
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Determination of chitin content in fungal cell wall: an alternative flow cytometric method.
Costa-de-Oliveira, Sofia; Silva, Ana P; Miranda, Isabel M; Salvador, Alexandre; Azevedo, Maria M; Munro, Carol A; Rodrigues, Acácio G; Pina-Vaz, Cidália
2013-03-01
The conventional methods used to evaluate chitin content in fungi, such as biochemical assessment of glucosamine release after acid hydrolysis or epifluorescence microscopy, are low throughput, laborious, time-consuming, and cannot evaluate a large number of cells. We developed a flow cytometric assay, efficient, and fast, based on Calcofluor White staining to measure chitin content in yeast cells. A staining index was defined, its value was directly related to chitin amount and taking into consideration the different levels of autofluorecence. Twenty-two Candida spp. and four Cryptococcus neoformans clinical isolates with distinct susceptibility profiles to caspofungin were evaluated. Candida albicans clinical isolate SC5314, and isogenic strains with deletions in chitin synthase 3 (chs3Δ/chs3Δ) and genes encoding predicted GlycosylPhosphatidylInositol (GPI)-anchored proteins (pga31Δ/Δ and pga62Δ/Δ), were used as controls. As expected, the wild-type strain displayed a significant higher chitin content (P relationship between chitin content and antifungal drug susceptibility phenotype was found, an association was established between the paradoxical growth effect in the presence of high caspofungin concentrations and the chitin content. This novel flow cytometry protocol revealed to be a simple and reliable assay to estimate cell wall chitin content of fungi. Copyright © 2013 International Society for Advancement of Cytometry.
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Flowcytometrisk diagnostik af hereditaer sfaerocytose
DEFF Research Database (Denmark)
Riley, Caroline H; Nikolajsen, Kirsten; Kjaersgaard, Erik
2009-01-01
defects in red cell cytoskeletal proteins. The dye eosin-5'-maleimide (EMA) binds to band three of the red cell membrane. The fluorescence intensity of EMA-labelled red cells can be quantified by flowcytometric analysis. Decreased fluorescence is found in patients with HS. We have evaluated this method...
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Cremer, Signe E; Krogh, Anne K H; Hedström, Matilda E K; Christiansen, Liselotte B; Tarnow, Inge; Kristensen, Annemarie T
2018-06-01
Platelet microparticles (PMPs) are subcellular procoagulant vesicles released upon platelet activation. In people with clinical diseases, alterations in PMP concentrations have been extensively investigated, but few canine studies exist. This study aims to validate a canine flow cytometric protocol for PMP quantification and to assess the influence of calcium on PMP concentrations. Microparticles (MP) were quantified in citrated whole blood (WB) and platelet-poor plasma (PPP) using flow cytometry. Anti-CD61 antibody and Annexin V (AnV) were used to detect platelets and phosphatidylserine, respectively. In 13 healthy dogs, CD61 + /AnV - concentrations were analyzed with/without a calcium buffer. CD61 + /AnV - , CD61 + /AnV + , and CD61 - /AnV + MP quantification were validated in 10 healthy dogs. The coefficient of variation (CV) for duplicate (intra-assay) and parallel (inter-assay) analyses and detection limits (DLs) were calculated. CD61 + /AnV - concentrations were higher in calcium buffer; 841,800 MP/μL (526,000-1,666,200) vs without; 474,200 MP/μL (278,800-997,500), P < .05. In WB, PMP were above DLs and demonstrated acceptable (<20%) intra-assay and inter-assay CVs in 9/10 dogs: 1.7% (0.5-8.9) and 9.0% (0.9-11.9), respectively, for CD61 + /AnV - and 2.4% (0.2-8.7) and 7.8% (0.0-12.8), respectively, for CD61 + /AnV + . Acceptable CVs were not seen for the CD61 - /AnV + MP. In PPP, quantifications were challenged by high inter-assay CV, overlapping DLs and hemolysis and lipemia interfered with quantification in 5/10 dogs. Calcium induced higher in vitro PMP concentrations, likely due to platelet activation. PMP concentrations were reliably quantified in WB, indicating the potential for clinical applications. PPP analyses were unreliable due to high inter-CV and DL overlap, and not obtainable due to hemolysis and lipemia interference. © 2018 American Society for Veterinary Clinical Pathology.
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Harper, Jason C; Polsky, Ronen; Wheeler, David R; Brozik, Susan M
2008-03-04
A versatile and simple method is introduced for formation of maleimide-functionalized surfaces using maleimide-activated aryl diazonium salts. We show for the first time electrodeposition of N-(4-diazophenyl)maleimide tetrafluoroborate on gold and carbon electrodes which was characterized via voltammetry, grazing angle FTIR, and ellipsometry. Electrodeposition conditions were used to control film thickness and yielded submonolayer-to-multilayer grafting. The resulting phenylmaleimide surfaces served as effective coupling agents for electrode functionalization with ferrocene and the redox-active protein cytochrome c. The utility of phenylmaleimide diazonium toward formation of a diazonium-activated conjugate, followed by direct electrodeposition of the diazonium-modified DNA onto the electrode surface, was also demonstrated. Effective electron transfer was obtained between immobilized molecules and the electrodes. This novel application of N-phenylmaleimide diazonium may facilitate the development of bioelectronic devices including biofuel cells, biosensors, and DNA and protein microarrays.
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Antifouling Thermoplastic Composites with Maleimide Encapsulated in Clay Nanotubes.
Fu, Ye; Gong, Congcong; Wang, Wencai; Zhang, Liqun; Ivanov, Evgenii; Lvov, Yuri
2017-09-06
An antifouling ethylene-vinyl acetate copolymer (EVA) coating with halloysite clay nanotubes loaded with maleimide (TCPM) is prepared. Such antifoulant encapsulation allowed for extended release of TCPM and a long-lasting, efficient protection of the coated surface against marine microorganisms proliferation. Halloysite also induces the composite's anisotropy due to parallel alignment of the nanotubes. The maleimide loaded halloysite incorporated into the polymer matrix allowed for 12-month release of the bacterial inhibitor preventing fouling; it is much longer than the 2-3 month protection when TCPM is directly admixed into EVA. The antifouling properties of the EVA-halloysite nanocomposites were tested by monitoring surface adhesion and proliferation of marine V. natriegens bacteria with SEM. As compared to the composite directly doped with TCPM-antifoulant, there were much less bacteria accumulated on the EVA-halloysite-TCPM coating after a 2-month exposure to seawater. Field tests at South China Sea marine station further confirmed the formulation efficiency. The doping of 28 wt % TCPM loaded halloysite drastically enhanced material antifouling property, which promises wide applications for protective marine coating.
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Novel β-cyclodextrin-eosin conjugates.
Benkovics, Gábor; Afonso, Damien; Darcsi, András; Béni, Szabolcs; Conoci, Sabrina; Fenyvesi, Éva; Szente, Lajos; Malanga, Milo; Sortino, Salvatore
2017-01-01
Eosin B (EoB) and eosin Y (EoY), two xanthene dye derivatives with photosensitizing ability were prepared in high purity through an improved synthetic route. The dyes were grafted to a 6-monoamino-β-cyclodextrin scaffold under mild reaction conditions through a stable amide linkage using the coupling agent 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium chloride. The molecular conjugates, well soluble in aqueous medium, were extensively characterized by 1D and 2D NMR spectroscopy and mass spectrometry. Preliminary spectroscopic investigations showed that the β-cyclodextrin-EoY conjugate retains both the fluorescence properties and the capability to photogenerate singlet oxygen of the unbound chromophore. In contrast, the corresponding β-cyclodextrin-EoB conjugate did not show either relevant emission or photosensitizing activity probably due to aggregation in aqueous medium, which precludes any response to light excitation.
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Novel β-cyclodextrin–eosin conjugates
Directory of Open Access Journals (Sweden)
Gábor Benkovics
2017-03-01
Full Text Available Eosin B (EoB and eosin Y (EoY, two xanthene dye derivatives with photosensitizing ability were prepared in high purity through an improved synthetic route. The dyes were grafted to a 6-monoamino-β-cyclodextrin scaffold under mild reaction conditions through a stable amide linkage using the coupling agent 4-(4,6-dimethoxy-1,3,5-triazin-2-yl-4-methylmorpholinium chloride. The molecular conjugates, well soluble in aqueous medium, were extensively characterized by 1D and 2D NMR spectroscopy and mass spectrometry. Preliminary spectroscopic investigations showed that the β-cyclodextrin–EoY conjugate retains both the fluorescence properties and the capability to photogenerate singlet oxygen of the unbound chromophore. In contrast, the corresponding β-cyclodextrin–EoB conjugate did not show either relevant emission or photosensitizing activity probably due to aggregation in aqueous medium, which precludes any response to light excitation.
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Naik, Sangeeta Mahableshwar; Anil, Arga Chandrashekar
2017-08-01
Immediate enumeration of phytoplankton is seldom possible. Therefore, fixation and subsequent storage are required for delayed analysis. This study investigated the influence of glutaraldehyde (GA) concentrations (0.25%, 0.5%, and 1%) and storage temperatures (-80°C LN2 , -80°C, -20°C, and 5°C) on Tetraselmis indica for flow cytometric analysis. Cell recovery, granularity, and membrane permeability were independent of GA concentration whereas cell size and chlorophyll autofluorescence were concentration dependent. After an initial cell loss (16-19%), no cell loss was observed when samples were stored at 5°C. Cell recovery was not influenced by storage temperature until 4months but later samples preserved at -80°C LN2 , -80°C, and -20°C resulted in ~41% cell loss. Although maximum cell recovery with minimal effect on cell integrity was obtained at 5°C, autofluorescence was retained better at -80°C LN2 and -80°C. This suggests that in addition to fixative, the choice of storage temperature is equally important. Thus for long-term preservation, especially to retain autofluorescence, the use of lower concentration (0.25%) of GA when stored at a lower temperature (-80°C LN2 and -80°C) while a higher concentration (1%) of GA when stored at a higher temperature (5°C) is recommended. Copyright © 2017 Elsevier B.V. All rights reserved.
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Energy Technology Data Exchange (ETDEWEB)
Wenz, F; Lohr, F; Rudat, V; Dietz, A; Flentje, M; Wannenmacher, M
1995-07-01
Purpose/Objective: Proliferation of surviving tumor cells during fractionated radiotherapy may limit tumor control, especially in rapidly proliferating tumors. It has been widely accepted, that this may play a major role in head and neck tumors. Several methods for the assessment of tumor proliferation have been developed, however, most of them are either laborious, invasive or potentially toxic. Today, the gold standard is the flow cytometric BrdUrd assay. We present a flow cytometric method for detection of PCNA, which is an intranuclear proliferation associated protein, in solid human head and neck tumors and how these data correlate with outcome. Materials and Methods: Pretherapeutic biopsies of 20 inoperable patients with squamous cell carcinoma of the head and neck (T3-4N2M0) were examined. The tissue was disaggregated with pepsin/HCl, antibody staining was performed using the clone PC10. Biparametric flow cytometry was performed after a FITC conjugated secondary antibody and propidiumjodine staining was applied. The PCNA-index (i.e. percentage PCNA-positive cells), the DNA-index and the S-phase fraction (SPF, euploid tumors only) were determined. The therapy consisted of combined accelerated-hyperfractionated radiochemotherapy (66 Gy in 5 wks, concomittant boost of 1.6 Gy/d in wks 4+5, Carboplatin in wks 1+5). The median follow-up time was 14 mths (5 - 28), the clinical partners (V.R., A.D.) were 'blinded' towards the PCNA-values. Results: 13 patients suffered from disease progession and 11 died. The actuarial median survival and disease free survival (DFS) were 14.4 and 10.7 mths, respectively. The PCNA-values ranged from 3.2 to 70% (median 9%), there were 7 aneuploid and 13 euploid tumors. SFP in the euploid tumors ranged from 4 to 14.5% (median 10.5%). Neither SFP nor ploidy had a significant influence on the outcome. The patients were divided according to their PCNA-value in higher (n=10) and lower (n=10) than the median. The survival and DFS were 13
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Vrieling, Engel G.; van de Poll, Willem H.; Vriezekolk, Gertie; Gieskes, Winfried W. C.
1997-05-01
The ichthyotoxic dinoflagellates Gyrodinium aureolum and Gymnodinium nagasakiense were cultured under different environmental conditions to test possible variability in immunochemical labelling intensity of cell-surface antigens using species-specific monoclonal antibodies. Variation of antigen abundance (which is directly related to labelling intensity) at the cell surface, determined by immuno-flow cytometry of cells labelled with FITC, appeared to be small but significant compared to control cultures. In general, a minor decrease in FIX fluorescence was recorded during exponential growth, followed by an increase during stationary growth. FITC fluorescence was correlated with cell size, shape and structure. This suggests a constant number of antigens per unit of cell surface. In all cultures, immunochemically labelled cells were distinguished clearly from unlabelled cells; immuno-flow cytometric identification is apparently not affected by growth conditions. Only at the end of the stationary growth phase in batch cultures did the FITC fluorescence values drop, which suggests that unhealthy, dying or lysing cells may either alter the composition of the cell surface or just fail to express the antigen.
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Directory of Open Access Journals (Sweden)
Fernando Fernández-Bañares
Full Text Available BACKGROUND & AIMS: An increase in CD3+TCRγδ+ and a decrease in CD3- intraepithelial lymphocytes (IEL is a characteristic flow cytometric pattern of celiac disease (CD with atrophy. The aim was to evaluate the usefulness of both CD IEL cytometric pattern and anti-TG2 IgA subepithelial deposit analysis (CD IF pattern for diagnosing lymphocytic enteritis due to CD. METHODS: Two-hundred and five patients (144 females who underwent duodenal biopsy for clinical suspicion of CD and positive celiac genetics were prospectively included. Fifty had villous atrophy, 70 lymphocytic enteritis, and 85 normal histology. Eight patients with non-celiac atrophy and 15 with lymphocytic enteritis secondary to Helicobacter pylori acted as control group. Duodenal biopsies were obtained to assess both CD IEL flow cytometric (complete or incomplete and IF patterns. RESULTS: Sensitivity of IF, and complete and incomplete cytometric patterns for CD diagnosis in patients with positive serology (Marsh 1+3 was 92%, 85 and 97% respectively, but only the complete cytometric pattern had 100% specificity. Twelve seropositive and 8 seronegative Marsh 1 patients had a CD diagnosis at inclusion or after gluten free-diet, respectively. CD cytometric pattern showed a better diagnostic performance than both IF pattern and serology for CD diagnosis in lymphocytic enteritis at baseline (95% vs 60% vs 60%, p = 0.039. CONCLUSIONS: Analysis of the IEL flow cytometric pattern is a fast, accurate method for identifying CD in the initial diagnostic biopsy of patients presenting with lymphocytic enteritis, even in seronegative patients, and seems to be better than anti-TG2 intestinal deposits.
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Spectroscopic determination of succinylcholine in dosage forms using eosin Y.
Ayad, Magda M; Belal, Fathalla; Hosney, Mervet M; Abo El Abass, Samah; Elsayed, Nora
2018-03-01
Two simple and sensitive analytical assay methods using spectrophotometry and spectrofluorimetry techniques were developed for the estimation of succinylcholine chloride (SUC) in pharmaceutical preparations. The suggested methods are based on the formation of an ion pair complex formed between the drug and eosin Y spectrophotometrically (Method I), or the suppressive effect of succinylcholine on the native fluorescence property of eosin Y (Method II). The spectrophotometric method (Method I) involves measuring the absorbance of the complex between succinylcholine and eosin Y at 550 nm in Britton Robinson buffer of pH 3. However, the spectrofluorimetric method (Method II) involves measuring the quenching effect of the studied drug on the native fluorescence property of eosin Y at the same pH at 550 nm after excitation at 480 nm. The absorbance versus concentration of the drug is rectilinear over the range of 0.5 to 15 μg/ml. The formation constant was 3.5 × 10 4 and the Gibb's free energy change was -2.5 × 10 4 J/mol. In Method II, the relative fluorescence intensity was directly proportional to SUC concentration over the range of 0.05 to 1 μg/ml. The proposed methods allowed a successful application to the estimation of succinylcholine ampoules. An explanation of the reaction pathway was postulated. Copyright © 2017 John Wiley & Sons, Ltd.
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Lignin-Based Materials Through Thiol-Maleimide "Click" Polymerization.
Buono, Pietro; Duval, Antoine; Averous, Luc; Habibi, Youssef
2017-03-09
In the present report an environmentally friendly approach to transforming renewable feedstocks into value-added materials is proposed. This transformation pathway was conducted under green conditions, without the use of solvents or catalyst. First, controlled modification of lignin, a major biopolymer present in wood and plants, was achieved by esterification with 11-maleimidoundecylenic acid (11-MUA), a derivative from castor oil that contains maleimide groups, following its transformation into 11-maleimidoundecanoyl chloride (11-MUC). Different degrees of substitution were achieved by using various amounts of the 11-MUC, leading to an efficient conversion of lignin hydroxy groups, as demonstrated by 1 H and 31 P NMR analyses. These fully biobased maleimide-lignin derivatives were subjected to an extremely fast (ca. 1 min) thiol-ene "click" polymerization with thiol-containing linkers. Aliphatic and aromatic thiol linkers bearing two to four thiol groups were used to tune the reactivity and crosslink density. The properties of the resulting materials were evaluated by swelling tests and thermal and mechanical analyses, which showed that varying the degree of functionality of the linker and the linker structure allowed accurate tailoring of the thermal and mechanical properties of the final materials, thus providing interesting perspectives for lignin in functional aromatic polymers. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.
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Energy Technology Data Exchange (ETDEWEB)
Buranda, T. [Univ. of New Mexico School of Medicine, Albuquerque, NM (United States)]|[Univ. of New Mexico, Albuquerque, NM (United States); Jones, G.M. [Univ. of New Mexico School of Medicine, Albuquerque, NM (United States); Nolan, J.P.; Keij, J. [Los Alamos National Labs., NM (United States); Lopez, G.P. [Univ. of New Mexico, Albuquerque, NM (United States); Sklar, L.A. [Univ. of New Mexico School of Medicine, Albuquerque, NM (United States)]|[Los Alamos National Lab., NM (United States)
1999-04-29
The authors have studied the binding of 5-((N-(5-(N-(6-(biotinoyl)amino)hexanoyl)amino)pentyl)thioureidyl)fluorescein (fluorescein biotin) to 6.2 {micro}m diameter, streptavidin-coated polystyrene beads using a combination of fluorimetric and flow cytometric methods. They have determined the average number of binding sites per bead, the extent of fluorescein quenching upon binding to the bead, and the association and dissociation kinetics. The authors estimate the site number to be {approx}1 million per bead. The binding of the fluorescein biotin ligand occurs in steps where the insertion of the biotin moiety into one receptor pocket is followed immediately by the capture of the fluorescein moiety by a neighboring binding pocket; fluorescence quenching is a consequence of this secondary binding. At high surface coverage, the dominant mechanism of quenching appears to be via the formation of nonfluorescent nearest-neighbor aggregates. At early times, the binding process is characterized by biphasic association and dissociation kinetics which are remarkably dependent on the initial concentration of the ligand. The rate constant for binding to the first receptor pocket of a streptavidin molecule is {approx}(1.3 {+-} 0.3) {times} 10{sup 7} 1{sup {minus}1} S{sup {minus}1}. The rate of binding of a second biotin may be reduced due to steric interference. The early time dissociative behavior is in sharp contrast to the typical stability associated with this system. The early time dissociative behavior is in sharp contrast to the typical stability associated with this system. The dissociation rate constant is as high as 0.05 s{sup {minus}1} shortly after binding, but decreases by 3 orders of magnitude after 3 h of binding. Potential sources for the time dependence of the dissociation rate constant are discussed.
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Directory of Open Access Journals (Sweden)
Xuan Duc Nguyen
2011-01-01
Full Text Available Transfusion-related acute lung injury (TRALI is a severe complication related with blood transfusion. TRALI has usually been associated with antibodies against leukocytes. The flow cytometric granulocyte immunofluorescence test (Flow-GIFT has been introduced for routine use when investigating patients and healthy blood donors. Here we describe a novel tool in the automation of the Flow-GIFT that enables a rapid screening of blood donations. We analyzed 440 sera from healthy female blood donors for the presence of granulocyte antibodies. As positive controls, 12 sera with known antibodies against anti-HNA-1a, -b, -2a; and -3a were additionally investigated. Whole-blood samples from HNA-typed donors were collected and the test cells isolated using cell sedimentation in a Ficoll density gradient. Subsequently, leukocytes were incubated with the respective serum and binding of antibodies was detected using FITC-conjugated antihuman antibody. 7-AAD was used to exclude dead cells. Pipetting steps were automated using the Biomek NXp Multichannel Automation Workstation. All samples were prepared in the 96-deep well plates and analyzed by flow cytometry. The standard granulocyte immunofluorescence test (GIFT and granulocyte agglutination test (GAT were also performed as reference methods. Sixteen sera were positive in the automated Flow-GIFT, while five of these sera were negative in the standard GIFT (anti—HNA 3a, n = 3; anti—HNA-1b, n = 1 and GAT (anti—HNA-2a, n = 1. The automated Flow-GIFT was able to detect all granulocyte antibodies, which could be only detected in GIFT in combination with GAT. In serial dilution tests, the automated Flow-GIFT detected the antibodies at higher dilutions than the reference methods GIFT and GAT. The Flow-GIFT proved to be feasible for automation. This novel high-throughput system allows an effective antigranulocyte antibody detection in a large donor population in order to prevent TRALI due to transfusion of
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Nguyen, Xuan Duc; Dengler, Thomas; Schulz-Linkholt, Monika; Klüter, Harald
2011-02-03
Transfusion-related acute lung injury (TRALI) is a severe complication related with blood transfusion. TRALI has usually been associated with antibodies against leukocytes. The flow cytometric granulocyte immunofluorescence test (Flow-GIFT) has been introduced for routine use when investigating patients and healthy blood donors. Here we describe a novel tool in the automation of the Flow-GIFT that enables a rapid screening of blood donations. We analyzed 440 sera from healthy female blood donors for the presence of granulocyte antibodies. As positive controls, 12 sera with known antibodies against anti-HNA-1a, -b, -2a; and -3a were additionally investigated. Whole-blood samples from HNA-typed donors were collected and the test cells isolated using cell sedimentation in a Ficoll density gradient. Subsequently, leukocytes were incubated with the respective serum and binding of antibodies was detected using FITC-conjugated antihuman antibody. 7-AAD was used to exclude dead cells. Pipetting steps were automated using the Biomek NXp Multichannel Automation Workstation. All samples were prepared in the 96-deep well plates and analyzed by flow cytometry. The standard granulocyte immunofluorescence test (GIFT) and granulocyte agglutination test (GAT) were also performed as reference methods. Sixteen sera were positive in the automated Flow-GIFT, while five of these sera were negative in the standard GIFT (anti-HNA 3a, n = 3; anti-HNA-1b, n = 1) and GAT (anti-HNA-2a, n = 1). The automated Flow-GIFT was able to detect all granulocyte antibodies, which could be only detected in GIFT in combination with GAT. In serial dilution tests, the automated Flow-GIFT detected the antibodies at higher dilutions than the reference methods GIFT and GAT. The Flow-GIFT proved to be feasible for automation. This novel high-throughput system allows an effective antigranulocyte antibody detection in a large donor population in order to prevent TRALI due to transfusion of blood products.
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ADSORPTION RATE CONSTANTS OF EOSIN IN HUMIN
anshar, andi muhammad
2015-01-01
Eosin is one of the dyes commonly used in the industry and has the potential to cause pollution of the water environment. The Eosin pollution treatment methods used in this study was the adsorption method using humin fraction obtained from the peat land comes from Kalimantan. From the research data showed that the adsorption of eosin in humin result of washing with HCl / HF optimum at pH 4 and a contact time of 60 minutes with the adsorption-order rate was 8,4 x 10-3 min-1
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Eosin-related fluorescence of acidophil pituitary cells
Friedman, H.; Friedman, I.V.C.; Mello, C.V.
1988-01-01
The examination of haematoxylin and eosin stained sections of normal and neoplastic pituitary glands under ultraviolet light illumination discloses fluorescence of acidophil cells. The distinction between prolactin and growth hormone-producing cells is not possible. Such fluorescence depends on previous eosin staining.
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Flow cytometric analysis of cell-surface and intracellular antigens in leukemia diagnosis.
Knapp, W; Strobl, H; Majdic, O
1994-12-15
New technology allows highly sensitive flow cytometric detection and quantitative analysis of intracellular antigens in normal and malignant hemopoietic cells. With this technology, the earliest stages of myeloid and lymphoid differentiation can easily and reliably be identified using antibodies directed against (pro-)myeloperoxidase/MPO, CD22 and CD3 antigens, respectively. Particularly for the analysis of undifferentiated acute myeloblastic leukemia (AML) cells, the immunological demonstration of intracellular MPO or its enzymatically inactive proforms is highly relevant, since other myeloid marker molecules such as CD33, CD13, or CDw65 are either not restricted to the granulomonocytic lineage or appear later in differentiation. By combining MPO staining with staining for lactoferrin (LF), undifferentiated cells can be distinguished from the granulomonocytic maturation compartment in bone marrow, since LF is selectively expressed from the myelocyte stage of differentiation onward. The list of informative intracellular antigens to be used in leukemia cell analysis will certainly expand in the near future. One candidate, intracellular CD68, has already been tested by us, and results are presented. Also dealt within this article are surface marker molecules not (as yet) widely used in leukemia cell analysis but with the potential to provide important additional information. Among them are the surface structures CD15, CD15s, CDw65, CD79a (MB-1), CD79b (B29), CD87 (uPA-R), and CD117 (c-kit).
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Kim, Eunju; Arul, Narayanasamy Sabari; Han, Jeong In
2016-06-01
This study reports the fabrication and investigation of the electrical properties of two types of conductive cotton yarns coated with eosin Y or eosin B functionalized reduced graphene (RGO) and bare graphene oxide (GO) using dip-coating method. The surface morphology of the conductive cotton yarn coated with reduced graphene oxide was observed by Scanning Electron Microscope (SEM). Due to the strong electrostatic attractive forces, the negatively charged surface such as the eosin Y functionalized reduced graphene oxide or bare GO can be easily coated to the positively charged polyethyleneimine (PEI) treated cotton yarn. The maximum current for the conductive cotton yarn coated with eosin Y functionalized RGO and bare GO with 20 cycles repetition of (5D + R) process was found to be 793.8 μA and 3482.8 μA. Our results showed that the electrical conductivity of bare GO coated conductive cotton yarn increased by approximately four orders of magnitude with the increase in the dipping cycle of (5D+R) process.
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Energy Technology Data Exchange (ETDEWEB)
Amartey, John K., E-mail: amarjk48@hotmail.co [Cyclotron and Radiopharmaceuticals Department, P.O. Box 3354, Riyadh 11211 (Saudi Arabia); Parhar, Ranjit S. [Biological and Medical Research Department, P.O. Box 3354, Riyadh 11211 (Saudi Arabia); Shi, Yufei [Genetics Department, King Faisal Specialist Hospital and Research Centre, P.O. Box 3354, Riyadh 11211 (Saudi Arabia); Al-Mohanna, Futwan [Biological and Medical Research Department, P.O. Box 3354, Riyadh 11211 (Saudi Arabia)
2011-01-15
A factor impeding the advancement of cell mediated therapy is the inability to track these cells in vivo by noninvasive techniques. It has been shown that cells express high levels of sulfhydryl groups. We sought to explore these groups to covalently label cells with radiolabeled maleimide derivatives. Two maleimide derivatives; N-[2-(2,5-dioxoazolinyl)ethyl](5-iodo(3-pyridyl))carboxamide and N-[2-(2,5-dioxoazolinyl)ethyl](3-iodophenyl)carboxamide ([{sup 125}I]-4 and [{sup 125}I]-8) were synthesized and radioiodinated. These compounds were evaluated for in vitro binding to neutrophils, endothelial and mesenchymal stem cells, and biodistribution of the radiolabeled stem cells in nude mice. These radiotracers were obtained in moderate to high radiochemical yields. Binding to cells were moderate (20-60%/10{sup 6} cells) and the label was retained, although washout (an average of 18-55%) was observed depending on the cell type and the tracer used. The labeled cells initially localized in well perfused organs and at a later time showed a general distribution as expected. The novel tracers labeled several cell types and shown that the stability of the label and viability of the cells were maintained in vitro and in vivo for a reasonable period and warrant further in vivo investigation.
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Flow Cytometric Detection of PrPSc in Neurons and Glial Cells from Prion-Infected Mouse Brains.
Yamasaki, Takeshi; Suzuki, Akio; Hasebe, Rie; Horiuchi, Motohiro
2018-01-01
In prion diseases, an abnormal isoform of prion protein (PrP Sc ) accumulates in neurons, astrocytes, and microglia in the brains of animals affected by prions. Detailed analyses of PrP Sc -positive neurons and glial cells are required to clarify their pathophysiological roles in the disease. Here, we report a novel method for the detection of PrP Sc in neurons and glial cells from the brains of prion-infected mice by flow cytometry using PrP Sc -specific staining with monoclonal antibody (MAb) 132. The combination of PrP Sc staining and immunolabeling of neural cell markers clearly distinguished neurons, astrocytes, and microglia that were positive for PrP Sc from those that were PrP Sc negative. The flow cytometric analysis of PrP Sc revealed the appearance of PrP Sc -positive neurons, astrocytes, and microglia at 60 days after intracerebral prion inoculation, suggesting the presence of PrP Sc in the glial cells, as well as in neurons, from an early stage of infection. Moreover, the kinetic analysis of PrP Sc revealed a continuous increase in the proportion of PrP Sc -positive cells for all cell types with disease progression. Finally, we applied this method to isolate neurons, astrocytes, and microglia positive for PrP Sc from a prion-infected mouse brain by florescence-activated cell sorting. The method described here enables comprehensive analyses specific to PrP Sc -positive neurons, astrocytes, and microglia that will contribute to the understanding of the pathophysiological roles of neurons and glial cells in PrP Sc -associated pathogenesis. IMPORTANCE Although formation of PrP Sc in neurons is associated closely with neurodegeneration in prion diseases, the mechanism of neurodegeneration is not understood completely. On the other hand, recent studies proposed the important roles of glial cells in PrP Sc -associated pathogenesis, such as the intracerebral spread of PrP Sc and clearance of PrP Sc from the brain. Despite the great need for detailed analyses
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Wood, Brent L; Arroz, Maria; Barnett, David; DiGiuseppe, Joseph; Greig, Bruce; Kussick, Steven J; Oldaker, Teri; Shenkin, Mark; Stone, Elizabeth; Wallace, Paul
2007-01-01
Immunophenotyping by flow cytometry has become standard practice in the evaluation and monitoring of patients with hematopoietic neoplasia. However, despite its widespread use, considerable variability continues to exist in the reagents used for evaluation and the format in which results are reported. As part of the 2006 Bethesda Consensus conference, a committee was formed to attempt to define a consensus set of reagents suitable for general use in the diagnosis and monitoring of hematopoietic neoplasms. The committee included laboratory professionals from private, public, and university hospitals as well as large reference laboratories that routinely operate clinical flow cytometry laboratories with an emphasis on lymphoma and leukemia immunophenotyping. A survey of participants successfully identified the cell lineage(s) to be evaluated for each of a variety of specific medical indications and defined a set of consensus reagents suitable for the initial evaluation of each cell lineage. Elements to be included in the reporting of clinical flow cytometric results for leukemia and lymphoma evaluation were also refined and are comprehensively listed. The 2006 Bethesda Consensus conference represents the first successful attempt to define a set of consensus reagents suitable for the initial evaluation of hematopoietic neoplasia. Copyright 2007 Clinical Cytometry Society.
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Grier, David D; Al-Quran, Samer Z; Cardona, Diana M; Li, Ying; Braylan, Raul C
2012-01-01
The diagnosis of B-cell lymphoma (BCL) is often dependent on the detection of clonal immunoglobulin (Ig) light chain expression. In some BCLs, the determination of clonality based on Ig light chain restriction may be difficult. The aim of our study was to assess the utility of flow cytometric analysis of surface Ig heavy chain (HC) expression in lymphoid tissues in distinguishing lymphoid hyperplasias from BCLs, and also differentiating various BCL subtypes. HC expression on B-cells varied among different types of hyperplasias. In follicular hyperplasia, IgM and IgD expression was high in mantle cells while germinal center cells showed poor HC expression. In other hyperplasias, B cell compartments were blurred but generally showed high IgD and IgM expression. Compared to hyperplasias, BCLs varied in IgM expression. Small lymphocytic lymphomas had lower IgM expression than mantle cell lymphomas. Of importance, IgD expression was significantly lower in BCLs than in hyperplasias, a finding that can be useful in differentiating lymphoma from reactive processes. PMID:22400070
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Giudice, Valentina; Feng, Xingmin; Lin, Zenghua; Hu, Wei; Zhang, Fanmao; Qiao, Wangmin; Ibanez, Maria Del Pilar Fernandez; Rios, Olga; Young, Neal S
2018-05-01
Oligoclonal expansion of CD8 + CD28 - lymphocytes has been considered indirect evidence for a pathogenic immune response in acquired aplastic anemia. A subset of CD8 + CD28 - cells with CD57 expression, termed effector memory cells, is expanded in several immune-mediated diseases and may have a role in immune surveillance. We hypothesized that effector memory CD8 + CD28 - CD57 + cells may drive aberrant oligoclonal expansion in aplastic anemia. We found CD8 + CD57 + cells frequently expanded in the blood of aplastic anemia patients, with oligoclonal characteristics by flow cytometric Vβ usage analysis: skewing in 1-5 Vβ families and frequencies of immunodominant clones ranging from 1.98% to 66.5%. Oligoclonal characteristics were also observed in total CD8 + cells from aplastic anemia patients with CD8 + CD57 + cell expansion by T-cell receptor deep sequencing, as well as the presence of 1-3 immunodominant clones. Oligoclonality was confirmed by T-cell receptor repertoire deep sequencing of enriched CD8 + CD57 + cells, which also showed decreased diversity compared to total CD4 + and CD8 + cell pools. From analysis of complementarity-determining region 3 sequences in the CD8 + cell pool, a total of 29 sequences were shared between patients and controls, but these sequences were highly expressed in aplastic anemia subjects and also present in their immunodominant clones. In summary, expansion of effector memory CD8 + T cells is frequent in aplastic anemia and mirrors Vβ oligoclonal expansion. Flow cytometric Vβ usage analysis combined with deep sequencing technologies allows high resolution characterization of the T-cell receptor repertoire, and might represent a useful tool in the diagnosis and periodic evaluation of aplastic anemia patients. (Registered at clinicaltrials.gov identifiers: 00001620, 01623167, 00001397, 00071045, 00081523, 00961064 ). Copyright © 2018 Ferrata Storti Foundation.
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Automation in high-content flow cytometry screening.
Naumann, U; Wand, M P
2009-09-01
High-content flow cytometric screening (FC-HCS) is a 21st Century technology that combines robotic fluid handling, flow cytometric instrumentation, and bioinformatics software, so that relatively large numbers of flow cytometric samples can be processed and analysed in a short period of time. We revisit a recent application of FC-HCS to the problem of cellular signature definition for acute graft-versus-host-disease. Our focus is on automation of the data processing steps using recent advances in statistical methodology. We demonstrate that effective results, on par with those obtained via manual processing, can be achieved using our automatic techniques. Such automation of FC-HCS has the potential to drastically improve diagnosis and biomarker identification.
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Technical discussions II - Flow cytometric analysis
Cunningham, A; Cid, A; Buma, AGJ
In this paper the potencial of flow cytometry as applied to the aquatic life sciences is discussed. The use of flow cytometry for studying the ecotoxicology of phytoplankton was introduced. On the other hand, the new flow cytometer EUROPA was presented. This is a multilaser machine which has been
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Retnakumari, Archana; Jayasimhan, Jasusri; Chandran, Parwathy; Menon, Deepthy; Nair, Shantikumar; Mony, Ullas; Koyakutty, Manzoor
2011-07-01
Protein stabilized gold nanoclusters (Au-NCs) are biocompatible, near-infrared (NIR) emitting nanosystems having a wide range of biomedical applications. Here, we report the development of a Au-NC based targeted fluorescent nano-bioprobe for the flow-cytometric detection of acute myeloid leukaemia (AML) cells. Au-NCs with ~ 25-28 atoms showing bright red-NIR fluorescence (600-750 nm) and average size of ~ 0.8 nm were prepared by bovine serum albumin assisted reduction-cum-stabilization in aqueous phase. The protein protected clusters were conjugated with monoclonal antibody against CD33 myeloid antigen, which is overexpressed in ~ 99.2% of the primitive population of AML cells, as confirmed by immunophenotyping using flow cytometry. Au-NC-CD33 conjugates having average size of ~ 12 nm retained bright fluorescence over an extended duration of ~ a year, as the albumin protein protects Au-NCs against degradation. Nanotoxicity studies revealed excellent biocompatibility of Au-NC conjugates, as they showed no adverse effect on the cell viability and inflammatory response. Target specificity of the conjugates for detecting CD33 expressing AML cells (KG1a) in flow cytometry showed specific staining of ~ 95.4% of leukaemia cells within 1-2 h compared to a non-specific uptake of ~ 8.2% in human peripheral blood cells (PBMCs) which are CD33low. The confocal imaging also demonstrated the targeted uptake of CD33 conjugated Au-NCs by leukaemia cells, thus confirming the flow cytometry results. This study demonstrates that novel nano-bioprobes can be developed using protein protected fluorescent nanoclusters of Au for the molecular receptor targeted flow cytometry based detection and imaging of cancer cells.
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International Nuclear Information System (INIS)
Retnakumari, Archana; Jayasimhan, Jasusri; Chandran, Parwathy; Menon, Deepthy; Nair, Shantikumar; Mony, Ullas; Koyakutty, Manzoor
2011-01-01
Protein stabilized gold nanoclusters (Au-NCs) are biocompatible, near-infrared (NIR) emitting nanosystems having a wide range of biomedical applications. Here, we report the development of a Au-NC based targeted fluorescent nano-bioprobe for the flow-cytometric detection of acute myeloid leukaemia (AML) cells. Au-NCs with ∼ 25-28 atoms showing bright red-NIR fluorescence (600-750 nm) and average size of ∼ 0.8 nm were prepared by bovine serum albumin assisted reduction-cum-stabilization in aqueous phase. The protein protected clusters were conjugated with monoclonal antibody against CD33 myeloid antigen, which is overexpressed in ∼ 99.2% of the primitive population of AML cells, as confirmed by immunophenotyping using flow cytometry. Au-NC-CD33 conjugates having average size of ∼ 12 nm retained bright fluorescence over an extended duration of ∼ a year, as the albumin protein protects Au-NCs against degradation. Nanotoxicity studies revealed excellent biocompatibility of Au-NC conjugates, as they showed no adverse effect on the cell viability and inflammatory response. Target specificity of the conjugates for detecting CD33 expressing AML cells (KG1a) in flow cytometry showed specific staining of ∼ 95.4% of leukaemia cells within 1-2 h compared to a non-specific uptake of ∼ 8.2% in human peripheral blood cells (PBMCs) which are CD33 low . The confocal imaging also demonstrated the targeted uptake of CD33 conjugated Au-NCs by leukaemia cells, thus confirming the flow cytometry results. This study demonstrates that novel nano-bioprobes can be developed using protein protected fluorescent nanoclusters of Au for the molecular receptor targeted flow cytometry based detection and imaging of cancer cells.
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Energy Technology Data Exchange (ETDEWEB)
Retnakumari, Archana; Jayasimhan, Jasusri; Chandran, Parwathy; Menon, Deepthy; Nair, Shantikumar; Mony, Ullas; Koyakutty, Manzoor, E-mail: manzoork@aims.amrita.edu, E-mail: ullasmony@aims.amrita.edu [Amrita Centre for Nanoscience and Molecular Medicine, Amrita Institute of Medical Science, Cochin 682 041 (India)
2011-07-15
Protein stabilized gold nanoclusters (Au-NCs) are biocompatible, near-infrared (NIR) emitting nanosystems having a wide range of biomedical applications. Here, we report the development of a Au-NC based targeted fluorescent nano-bioprobe for the flow-cytometric detection of acute myeloid leukaemia (AML) cells. Au-NCs with {approx} 25-28 atoms showing bright red-NIR fluorescence (600-750 nm) and average size of {approx} 0.8 nm were prepared by bovine serum albumin assisted reduction-cum-stabilization in aqueous phase. The protein protected clusters were conjugated with monoclonal antibody against CD33 myeloid antigen, which is overexpressed in {approx} 99.2% of the primitive population of AML cells, as confirmed by immunophenotyping using flow cytometry. Au-NC-CD33 conjugates having average size of {approx} 12 nm retained bright fluorescence over an extended duration of {approx} a year, as the albumin protein protects Au-NCs against degradation. Nanotoxicity studies revealed excellent biocompatibility of Au-NC conjugates, as they showed no adverse effect on the cell viability and inflammatory response. Target specificity of the conjugates for detecting CD33 expressing AML cells (KG1a) in flow cytometry showed specific staining of {approx} 95.4% of leukaemia cells within 1-2 h compared to a non-specific uptake of {approx} 8.2% in human peripheral blood cells (PBMCs) which are CD33{sup low}. The confocal imaging also demonstrated the targeted uptake of CD33 conjugated Au-NCs by leukaemia cells, thus confirming the flow cytometry results. This study demonstrates that novel nano-bioprobes can be developed using protein protected fluorescent nanoclusters of Au for the molecular receptor targeted flow cytometry based detection and imaging of cancer cells.
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Hadjichristidis, Nikolaos; Alkayal, Nazeeha
2015-01-01
A novel strategy towards well-defined polymethylene-based co/terpolymers, by combining anthracene/maleimide Diels-Alder reaction with polyhomologation, is presented. For the synthesis of diblock copolymers the following approach was applied: a
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Development of bioconjugated dye-doped poly(styrene-co-maleimide) nanoparticles as a new bioprobe
CSIR Research Space (South Africa)
Swanepoel, A
2015-02-01
Full Text Available -1 Journal of Materials Chemistry B Development of bioconjugated dye-doped poly(styrene-co- maleimide) nanoparticles as a new bioprobe A. Swanepoel, I. du Preez, T. Mahlangu, A. Chetty and B. Klumperman Abstract Fluorescent dye-doped poly...
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Directory of Open Access Journals (Sweden)
B. D. Kouame
2014-01-01
Full Text Available Background: The surgical management of giant omphalocele is a surgical challenge with high mortality and morbidity in our country due to the absence of neonatal resuscitation. This study evaluates conservative management of giant omphalocele with dissodic 2% aqueous eosin. Materials and Methods: In the period from January 1997 to December 2012, giant omphaloceles were treated with dissodic 2% aqueous eosin. The procedure consisted of twice a day application of dissodic 2% aqueous eosin (sterile solution for topical application on the omphalocele sac. The procedure was taught to the mother to continue at home with an outpatient follow-up to assess epithelialization. We studied the duration of the hospital stay, the learning curve of the procedure by the mother, the complications, the duration and the percentage of complete epithelialization and the mortality. Results: A total of 173 giant omphaloceles had a conservative treatment with dissodic 2% aqueous eosin. The average hospital stay was 21 ± 6 days. The learning curve by the mother of the procedure was 10 ± 3 days. Complications of treatment were intestinal functional occlusion 22% and omphalocele sac infection 18%. The complete epithelialization of the omphaloceles sac after application of dissodic 2% aqueous eosin was 68.5%. Mortality was observed in 25.5%. Conclusion: Conservative treatment of giant omphaloceles by dissodic 2% aqueous eosin is a simple, efficient and a good alternative to surgery. The mother can easily learn its procedure which reduces the duration of hospital stay.
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Eosin blue dye based poly(methacrylate) films for data storage
Sankar, Deepa; Palanisamy, P. K.; Manickasundaram, S.; Kannan, P.
2006-06-01
Eosin dye based poly(methacrylates) with variation in the number of methylene spacers have been prepared by free radical polymerization process. The utility of the polymers for high-density optical data storage using holography has been studied by grating formation with the 514.5 nm line of the Argon ion laser as source. The influence of various parameters on the diffraction efficiency of the polymers has been studied. The effect of increase in the number of methylene spacers, hooked to the eosin blue dye, on the diffraction efficiency of the grating formed has also been discussed. Optical microscopic observations showing grating formation in the polymers have also been presented.
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Yokosuka, Tomoko; Goto, Hiroaki; Fujii, Hisaki; Naruto, Takuya; Takeuchi, Masanobu; Tanoshima, Reo; Kato, Hiromi; Yanagimachi, Masakatsu; Kajiwara, Ryosuke; Yokota, Shumpei
2013-12-01
The purpose of the study is to establish a simple and relatively inexpensive flow cytometric chemosensitivity assay (FCCA) for leukemia to distinguish leukemic blasts from normal leukocytes in clinical samples. We first examined whether the FCCA with the mitochondrial membrane depolarization sensor, 5, 50, 6, 60-tetrachloro-1, 10, 3, 30 tetraethyl benzimidazolo carbocyanine iodide (JC-1), could detect drug-induced apoptosis as the conventional FCCA by annexin V/7-AAD detection did and whether it was applicable in the clinical samples. Second, we compared the results of the FCCA for prednisolone (PSL) with clinical PSL response in 18 acute lymphoblastic leukemia (ALL) patients to evaluate the reliability of the JC-1 FCCA. Finally, we performed the JC-1 FCCA for bortezomib (Bor) in 25 ALL or 11 acute myeloid leukemia (AML) samples as the example of the clinical application of the FCCA. In ALL cells, the results of the JC-1 FCCA for nine anticancer drugs were well correlated with those of the conventional FCCA using anti-annexin V antibody (P < 0.001). In the clinical samples from 18 children with ALL, the results of the JC-1 FCCA for PSL were significantly correlated with the clinical PSL response (P = 0.005). In ALL samples, the sensitivity for Bor was found to be significantly correlated with the sensitivity for PSL (P = 0.005). In AML samples, the Bor sensitivity was strongly correlated with the cytarabine sensitivity (P = 0.0003). This study showed the reliability of a relatively simple and the FCCA using JC-1, and the possibility for the further clinical application.
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Kerényi, Adrienne; Beke Debreceni, Ildikó; Oláh, Zsolt; Ilonczai, Péter; Bereczky, Zsuzsanna; Nagy, Béla; Muszbek, László; Kappelmayer, János
2017-09-01
Heparin-induced thrombocytopenia (HIT) is a severe side effect of heparin treatment caused by platelet activating IgG antibodies generated against the platelet factor 4 (PF4)-heparin complex. Thrombocytopenia and thrombosis are the leading clinical symptoms of HIT. The clinical pretest probability of HIT was evaluated by the 4T score system. Laboratory testing of HIT was performed by immunological detection of antibodies against PF4-heparin complex (EIA) and two functional assays. Heparin-dependent activation of donor platelets by patient plasma was detected by flow cytometry. Increased binding of Annexin-V to platelets and elevated number of platelet-derived microparticles (PMP) were the indicators of platelet activation. EIA for IgG isotype HIT antibodies was performed in 405 suspected HIT patients. Based on negative EIA results, HIT was excluded in 365 (90%) of cases. In 40 patients with positive EIA test result functional tests were performed. Platelet activating antibodies were detected in 17 cases by Annexin V binding. PMP count analysis provided nearly identical results. The probability of a positive flow cytometric assay result was higher in patients with elevated antibody titer. 71% of patients with positive EIA and functional assay had thrombosis. EIA is an important first line laboratory test in the diagnosis of HIT; however, HIT must be confirmed by a functional test. Annexin V binding and PMP assays using flow cytometry are functional HIT tests convenient in a clinical diagnostic laboratory. The positive results of functional assays may predict the onset of thrombosis. © 2016 International Clinical Cytometry Society. © 2016 International Clinical Cytometry Society.
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Smarda, P; Stancík, D
2006-01-01
Ploidy levels and chromosome numbers for 24 species of Festuca L. from 29 sites in Bolivia, Colombia, Ecuador and Venezuela are given. The ploidy level of 22 species is reported for the first time. A higher proportion of tetraploids in northern South America and the high frequency of polyploids in the whole continent are documented. In combination with chromosome counts, ploidy level was determined using flow cytometry in 4- to 5 1/2 -year-old herbarium specimens and mature caryopses. Flow cytometric determination from seeds was more reliable than determination from herbarium specimens. In herbarium specimens, the youngest, fresh green leaves, still hidden in sheaths, seem to be most suitable for cytometric determination. In old, brownish leaves, or poorly preserved herbarium specimens, the degradation of DNA signal in flow histograms was documented. DNA content measured in seeds was always higher than that measured in herbarium specimens, which may be caused by the presence of different cytosolic compounds. Differences of about 15% in relative DNA content of F. sodiroana and F. vaginalis was found in simultaneous measurements in seeds.
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Nonlinear multicontrast microscopy of hematoxylin-and-eosin-stained histological sections
Tuer, Adam; Tokarz, Danielle; Prent, Nicole; Cisek, Richard; Alami, Jennifer; Dumont, Daniel J.; Bakueva, Ludmila; Rowlands, John; Barzda, Virginijus
2010-03-01
Imaging hematoxylin-and-eosin-stained cancerous histological sections with multicontrast nonlinear excitation fluorescence, second- and third-harmonic generation (THG) microscopy reveals cellular structures with extremely high image contrast. Absorption and fluorescence spectroscopy together with second hyperpolarizability measurements of the dyes shows that strong THG appears due to neutral hemalum aggregation and is subsequently enhanced by interaction with eosin. Additionally, fluorescence lifetime imaging microscopy reveals eosin fluorescence quenching by hemalums, showing better suitability of only eosin staining for fluorescence microscopy. Multicontrast nonlinear microscopy has the potential to differentiate between cancerous and healthy tissue at a single cell level.
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International Nuclear Information System (INIS)
Lim, Eng Liang; Yap, Chi Chin; Yahaya, Muhammad; Salleh, Muhamad Mat
2013-01-01
This paper reports the effect of Eosin-Y coating concentration on the performance of inverted type hybrid organic solar cell based on ZnO nanorod arrays and poly(3-hexylthiophene-2,5-diyl) (P3HT). The Eosin-Y solution with concentrations of 0.05, 0.2, 2.0 and 5.0 mM was spin-coated onto the ZnO nanorod arrays grown on the fluorine-doped tin oxide glass substrate. The P3HT film was then spin-coated onto Eosin-Y-coated ZnO nanorod arrays, followed by deposition of silver (Ag) as anode using magnetron sputtering technique. The short circuit current density increased with the Eosin-Y coating concentration up to 0.2 mM, after which it started to decrease, mainly due to the aggregation of Eosin-Y which reduced the charge extraction from P3HT to ZnO. Meanwhile, the open circuit voltage increased with the Eosin-Y coating concentration, indicating reduced back charge recombination of electron on the ZnO and hole on the P3HT, as well as reduced leakage current through the direct contact between the ZnO nanorods and the Ag metal contact. The power conversion efficiency of the device with the optimum coating concentration was approximately eight times higher than that without Eosin-Y modification. (paper)
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Lim, Eng Liang; Yap, Chi Chin; Yahaya, Muhammad; Mat Salleh, Muhamad
2013-04-01
This paper reports the effect of Eosin-Y coating concentration on the performance of inverted type hybrid organic solar cell based on ZnO nanorod arrays and poly(3-hexylthiophene-2,5-diyl) (P3HT). The Eosin-Y solution with concentrations of 0.05, 0.2, 2.0 and 5.0 mM was spin-coated onto the ZnO nanorod arrays grown on the fluorine-doped tin oxide glass substrate. The P3HT film was then spin-coated onto Eosin-Y-coated ZnO nanorod arrays, followed by deposition of silver (Ag) as anode using magnetron sputtering technique. The short circuit current density increased with the Eosin-Y coating concentration up to 0.2 mM, after which it started to decrease, mainly due to the aggregation of Eosin-Y which reduced the charge extraction from P3HT to ZnO. Meanwhile, the open circuit voltage increased with the Eosin-Y coating concentration, indicating reduced back charge recombination of electron on the ZnO and hole on the P3HT, as well as reduced leakage current through the direct contact between the ZnO nanorods and the Ag metal contact. The power conversion efficiency of the device with the optimum coating concentration was approximately eight times higher than that without Eosin-Y modification.
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Daubeuf, François; Becker, Julien; Aguilar-Pimentel, Juan Antonio; Ebel, Claudine; Hrabě de Angelis, Martin; Hérault, Yann; Frossard, Nelly
2017-06-19
The cell composition of bronchoalveolar lavage fluid (BAL) is an important indicator of airway inflammation. It is commonly determined by cytocentrifuging leukocytes on slides, then staining, identifying, and counting them as eosinophils, neutrophils, macrophages, or lymphocytes according to morphological criteria under light microscopy, where it is not always easy to distinguish macrophages from lymphocytes. We describe here a one-step, easy-to-use, and easy-to-customize 8-color flow cytometric method for performing differential cell count and comparing it to morphological counts on stained cytospins. This method identifies BAL cells by a simultaneous one-step immunolabeling procedure using antibodies to identify T cells, B cells, neutrophils, eosinophils, and macrophages. Morphological analysis of flow-sorted cell subsets is used to validate this protocol. An important advantage of this basic flow cytometry protocol is the ability to customize it by the addition of antibodies to study receptor expression at leukocyte cell surfaces and identify subclasses of inflammatory cells as needed. © 2017 by John Wiley & Sons, Inc. Copyright © 2017 John Wiley & Sons, Inc.
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Cytometric approaches to biological dosimetry
International Nuclear Information System (INIS)
Burger, G.
1983-01-01
Automatic cytometric techniques for detecting chromosomal aberrations are being tested but will not be used in routine examinations for some time to come. Automatic micronuclei counts are more promising but not sufficiently sensitive in the low dose range ( [de
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Quirks of dye nomenclature. 10. Eosin Y and its close relatives.
Cooksey, C J
2018-02-08
The long history of eosin Y, eosin B and the methyl and ethyl eosins is recounted as well as their synthesis, the variety of their molecular species and some of the myriad applications of these dyes. Chromatographic techniques are described that reveal the purity or lack of it in commercial samples. Toxicological studies are discussed that suggest that the eosins are virtually non toxic, but efforts to remove them from the environment imply that there may be some risk.
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Directory of Open Access Journals (Sweden)
Jesús M. Egido
1997-12-01
Full Text Available We report a rapid method for the flow cytometric quantitation of phagocytosis in heparinized complete peripherial blood (HCPB, using commercially available phycoerythrin-conjugated latex particles of 1µm diameter. The method is faster and shows greater reproducibility than Bjerknes' (1984 standard technique using propidium iodide-stained Candida albicans, conventionally applied to the leukocytic layer of peripherial blood but here modified for HCPB. We also report a modification of Bjerknes' Intracellular Killing Test to allow its application to HCPB.Se da cuenta de un método rápido para la cuantización del flujo citométrico de la fagocitosis en sangre periférica completamente heparinizada (HCPB, mediante la utilización de partículas de látex phycoerythrin-conjugadas de 1µm de diámetro disponibles comercialmente. El método es más rápido y presenta mayor reproducibilidad que la técnica estandar de Bjerknes' (1984 utilizando propidium iodide-teñida Candida albicans, aplicada convencionalmente a la capa leucocitica de sangre periférica pero modificada por HCPB. Tambien damos cuenta de una modificación de Bjerknes' Intracellular Killing Test para permitir su aplicación a HCPB.
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Reyes-Núñez, Virginia; Galo-Hooker, Evelyn; Pérez-Romano, Beatriz; Duque, Ricardo E; Ruiz-Arguelles, Alejandro; Garcés-Eisele, Javier
2018-01-01
The aim of this work was to simultaneously use multiplex ligation-dependent probe amplification (MLPA) assay and flow cytometric DNA ploidy analysis (FPA) to detect aneuploidy in patients with newly diagnosed acute leukemia. MLPA assay and propidium iodide FPA were used to test samples from 53 consecutive patients with newly diagnosed acute leukemia referred to our laboratory for immunophenotyping. Results were compared by nonparametric statistics. The combined use of both methods significantly increased the rate of detection of aneuploidy as compared to that obtained by each method alone. The limitations of one method are somehow countervailed by the other and vice versa. MPLA and FPA yield different yet complementary information concerning aneuploidy in acute leukemia. The simultaneous use of both methods might be recommended in the clinical setting. © 2017 International Clinical Cytometry Society. © 2017 International Clinical Cytometry Society.
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Suzuki, Hiroshi; Toyooka, Tatsushi; Ibuki, Yuko
2007-04-15
Many classes of nanoparticles have been synthesized and widely applied, however, there is a serious lack of information concerning their effects on human health and the environment. Considering that their use will increase, accurate and cost-effective measurement techniques for characterizing "nanotoxicity" are required. One major toxicological concern is that nanoparticles are easily taken up in the human body. In this study, we developed a method of evaluating the uptake potential of nanosized particles using flow cytometric light scatter. Suspended titanium dioxide (TiO2) particles (5, 23, or 5000 nm) were added to Chinese hamster ovary cells. Observation by confocal laser scanning microscopy showed that the TiO2 particles easily moved to the cytoplasm of the cultured mammalian cells, not to the nucleus. The intensity of the side-scattered light revealed that the particles were taken up in the cells dose-, time-, and size-dependently. In addition, surface-coating of TiO2 particles changed the uptake into the cells, which was accurately reflected in the intensity of the side-scattered light. The uptake of other nanoparticles such as silver (Ag) and iron oxide (Fe3O4) also could be detected. This method could be used for the initial screening of the uptake potential of nanoparticles as an index of "nanotoxicity".
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Study on the fluorescence resonance energy transfer between CdS quantum dots and Eosin Y.
Yan, Zhengyu; Zhang, Zhengwei; Yu, Yan; Chen, Jianqiu
2015-03-01
Water-soluble CdS quantum dots (QDs) were prepared using mercaptoacetic acid (TGA) as the stabilizer in an aqueous system. A fluorescence resonance energy transfer (FRET) system was constructed between water-soluble CdS QDs (donor) and Eosin Y (acceptor). Several factors that impacted the fluorescence spectra of the FRET system, such as pH (3.05-10.10), concentration of Eosin Y (2-80 mg/L) and concentration of CdS QDs (2-80 mg/L), were investigated and refined. Donor-to-acceptor ratios, the energy transfer efficiency (E) and the distance (r) between CdS QDs and Eosin Y were obtained. The results showed that a FRET system could be established between water-soluble CdS QDs and Eosin Y at pH 5.0; donor-to-acceptor ratios demonstrated a 1: 8 proportion of complexes; the energy transfer efficiency (E) and the distance (r) between the QDs and Eosin Y were 20.07% and 4.36 nm,respectively. Copyright © 2014 John Wiley & Sons, Ltd.
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N-(m-[125I]iodophenyl)maleimide: an agent for high yield radiolabeling of antibodies
International Nuclear Information System (INIS)
Khawli, L.A.; Van den Abbeele, A.D.; Kassis, A.I.
1992-01-01
In an effort to radiolabel antibodies, N-(m-[ 125 I]iodophenyl)maleimide (m-[ 125 I]IPM) was prepared by the demetallation of an N-[m-tri-(n-butyl)stannylphenyl]maleimide intermediate. The unlabeled intermediate was synthesized in ≥ 75% yield using a palladium catalyzed reaction of hexabutylditin with m-bromoaniline, followed by reaction with maleic anhydride and ring annulation. All products were confirmed by NMR and elemental analysis. Labeling with 125 I was carried out in a biphasic mixture containing chloramine-T (radiochemical yield ≥ 70%). Rabbit IgG modified with the heterobifunctional crosslinking agent N-succinimidyl-3-(2-pyridyldithio)propionate (SPDP) and bovine serum albumin were conjugated with m-[ 125 I]IPM (yield: 40 and 80%, respectively). In addition, m-[ 125 I]IPM was conjugated to rabbit IgG subunits (HL) in 70% yield. The in vitro stability of the radiolabeled proteins in serum showed < 1% deiodination over 24h. (author)
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Mechanisms involved in the chemical inhibition of the Eosin-sensitized photooxidation of trypsin
Energy Technology Data Exchange (ETDEWEB)
Rizzuto, F.; Spikes, J.D.
1975-01-01
A large series of compounds was screened for ability to protect trypsin from eosin-sensitized photodynamic inactivation. Eosin-sensitized photooxidation reactions of this type typically proceed via the triplet state of the dye and often involve singlet state oxygen as the oxidizing entity. In order to determine the mechanisms by which trypsin is protected from photoinactivation, a number of good protective agents (inhibitors) and some non-protective agents were selected for more detailed flash photolysis studies. Good inhibitors such as p-phenylenediamine, n-propyl gallate, serotonin creatinine sulfate and p-toluenediamine competed efficiently with oxygen and with trypsin for reaction with the triplet state of eosin. The inhibitors were shown to quench triplet eosin to the ground state and/or reduce triplet eosin to form the semireduced eosin radical and an oxidized form of the inhibitor. In the latter case, oxidized inhibitor could react by a reverse electron transfer reaction with the semireduced eosin radical to regenerate ground state eosin and the inhibitor. The good inhibitors also competed effectively with trypsin for oxidation by semioxidized eosin, thus giving another possible protective mechanism. Non-inhibitors such as halogen ions and the paramagnetic ions Co/sup + +/, Cu/sup + +/ and Mn/sup + +/ reacted only slowly with triplet and with semioxidized eosin. The primary pathway for the eosin-sensitized photooxidation of trypsin at pH 8.0 involved singlet oxygen, although semioxidized eosin may also participate.
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Valle, Julio; Morgado, José Mario T; Ruiz-Martín, Juan; Guardiola, Antonio; Lopes-Nogueras, Miriam; García-Vela, Almudena; Martín-Sacristán, Beatriz; Sánchez-Muñoz, Laura
2017-10-01
Diagnosis of celiac disease is difficult when the combined results of serology and histology are inconclusive. Studies using flow cytometry of intraepithelial lymphocytes (IELs) have found that celiac patients have increased numbers of γδ IELs, along with a decrease in CD3-CD103 + IELs. The objective of this article is to assess the role of flow cytometric analysis of IELs in the diagnosis of celiac disease in difficult cases. A total of 312 patients with suspicion of celiac disease were included in the study. Duodenal biopsy samples were used for histological assessment and for flow cytometric analysis of IELs. In 46 out of 312 cases (14.7%) the combination of serology and histology did not allow the confirmation or exclusion of celiac disease. HLA typing had been performed in 42 of these difficult cases. Taking into account HLA typing and the response to a gluten-free diet, celiac disease was excluded in 30 of these cases and confirmed in the remaining 12. Flow cytometric analysis of IELs allowed a correct diagnosis in 39 out of 42 difficult cases (92.8%) and had a sensitivity of 91.7% (95% CI: 61.5% to 99.8%) and a specificity of 93.3% (95% CI: 77.9% to 99.2%) for the diagnosis of celiac disease in this setting. Flow cytometric analysis of IELs is useful for the diagnosis of celiac disease in difficult cases.
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Flow cytometry and integrated imaging
Directory of Open Access Journals (Sweden)
V. Kachel
2000-06-01
Full Text Available It is a serious problem to relate the results of a flow cytometric analysis of a marine sample to different species. Images of particles selectively triggered by the flow cytometric analysis and picked out from the flowing stream give a valuable additional information on the analyzed organisms. The technical principles and problems of triggered imaging in flow are discussed, as well as the positioning of the particles in the plane of focus, freezing the motion of the quickly moving objects and what kinds of light sources are suitable for pulsed illumination. The images have to be stored either by film or electronically. The features of camera targets and the memory requirements for storing the image data and the conditions for the triggering device are shown. A brief explanation of the features of three realized flow cytometric imaging (FCI systems is given: the Macro Flow Planktometer built within the EUROMAR MAROPT project, the Imaging Module of the European Plankton Analysis System, supported by the MAST II EurOPA project and the most recently developed FLUVO VI universal flow cytometer including HBO 100- and laser excitation for fluorescence and scatter, Coulter sizing as well as bright field and and phase contrast FCI.
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Synthesis and 1,3-Dipolar Cycloaddition Reactions of Chiral Maleimides
Directory of Open Access Journals (Sweden)
Lubor Fisera
1997-02-01
Full Text Available New routes to the synthesis of various novel chiral maleimides are described. The oxabicyclic anhydride 2 readily available exo-Diels-Alder adduct of furan and maleic anhydride was used as a vehicle, which in turn reacted with hydrochlorides of amino acids 3a-f in the presence of Et3N with release of furan to give the requisite novel chiral imides 4a-f in good to moderate yields. The stereoselectivity of 1,3-dipolar cycloaddition of nitrile oxides with prepared chiral imides 4a-f is investigated.
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Dasgupta, Alakananda; Mahapatra, Manoranjan; Saxena, Renu
2013-04-01
This study analyzed the frequency of regulatory T cells (Tregs) in chronic lymphocytic leukemia (CLL) by multiparameter flow cytometric immunophenotyping. Patients showed significantly increased frequencies of Tregs as compared to controls, a significantly higher percentage than that identified by previous studies, possibly indicating a different prognosis of CLL in different parts of the world and, more precisely, a worse prognosis of CLL in the Indian population. A higher frequency of Tregs was also seen in advanced stage of disease with significantly reduced frequencies of Tregs in patients with CLL after chemotherapy. A significant proportion of CD127low/-FoxP3+ Tregs expressed only low levels of CD25. Thus, CD127 appears to be a better marker than CD25 for the identification of CD4+FoxP3+ T cells as potential Tregs. Our results suggest that the specificity and sensitivity of CD4+CD127low/- cells are comparable to those of CD4+FoxP3+, which is the gold standard, and can be used as an alternative. This novel flow cytometric antibody panel with fewer number of antibodies is cost-effective and can be used to enumerate Tregs in resource-limited settings.
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Fluorescent eosin probe in investigations of structural changes in glycated proteins
Pravdin, A. B.; Kochubey, V. I.; Mel'Nikov, A. G.
2010-08-01
The possibility of using the luminescent-kinetic probe method to investigate structural changes in bovine serum albumin (BSA) upon nonenzymatic thermal glycation is studied. An increase in the glycation time lead to a decrease in the intensity of the probe (eosin) fluorescence and to a long-wavelength shift of its maximum, as well as to an increase in the eosin phosphorescence intensity, which indicates that eosin binds to hydrophobic regions of protein at any times of incubation of BSA with glucose. From a decrease in the rate constant of the triplet-triplet energy transfer between the donor (eosin) and acceptor (anthracene) bound to proteins, it is found that the changes observed in the spectral characteristics of eosin are caused by structural changes in albumin globules as a result of glycosylation.
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Capillary Electrophoresis Artifact Due to Eosin
Murphy, Kathleen M.; Berg, Karin D.; Geiger, Tanya; Hafez, Michael; Flickinger, Katie A.; Cooper, Lisa; Pearson, Patrick; Eshleman, James R.
2005-01-01
Capillary electrophoresis (CE) is a commonly used tool in the analysis of fluorescently labeled PCR amplification products. We have identified a CE artifact caused by the tissue stain eosin that can complicate the interpretation of CE data. The artifact was detected during routine analysis of a DNA sample isolated from a formalin-fixed, paraffin-embedded tissue sample considered histologically suspicious for a B-cell neoplasm. A standard clinical PCR and CE assay for immunoglobulin heavy chain (IGH) gene rearrangement revealed a weak polyclonal population of rearranged IGH genes and a 71 base peak suspicious for IGH clonality. The spectral properties of the 71 base peak were unusual in that although the dominant fluorescence of the peak was blue, it also fluoresced in green and yellow (blue>green>yellow), raising the suspicion that the peak might represent an artifact. CE analysis of the genomic DNA sample without PCR amplification demonstrated the presence of the 71 base peak, suggesting that the artifact was caused by a contaminant within the DNA sample itself. We demonstrate that eosin, which was used to stain the formalin-fixed tissue during processing, yields a discrete 71 base peak of similar morphology to the contaminant peak on CE analysis. The data suggest that eosin in the fixed tissue was not completely eliminated during nucleic acid extraction, resulting in the artifact peak. We discuss the implications of this potentially common contaminant on the interpretation of CE data and demonstrate that artifacts caused by eosin can be avoided by using more stringent DNA purification steps. Histological dyes may fluoresce, and artifacts from them should be considered when primary peaks contain additional underlying peaks of other colors. PMID:15681487
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Luminescent Eosin Y–SiO2 hybrid nano and microrods prepared by sol–gel template method
International Nuclear Information System (INIS)
Secu, M.; Secu, C.E.; Sima, M.; Negrea, R.F.; Bartha, C.; Dinescu, M.; Damian, V.
2013-01-01
Sol–gel chemistry within the pores of a polycarbonate template membrane was used for the preparation of Eosin Y–SiO 2 hybrid nano- and microrods, using tetraethylorthosilicate [TEOS, Si(OC 2 H 5 ) 4 ] as the precursor in the presence of trifluoroacetic acid (TFA) catalyst. The ethanolic solution of Eosin-Y was added to the silica sol to trap dye molecules inside the SiO 2 gel network during the gelation. Structural and morphological characterization using scanning electron microscopy (SEM) and luminescence microscopy have shown the formation of rods with 200 nm and 1.2 μm diameter and about 30 μm length, exhibiting luminescence properties. Spectroscopic characterization has shown that the luminescence is due to Eosin-Y molecule in the xerogel porous network, surrounded by a solvation shell given mainly by the water. -- Highlights: • Sol–gel template method was used to prepare Eosin Y–SiO 2 hybrid rods-type structures. • Morphological characterization has shown nano- and microrods with luminescent properties. • Luminescence is due to Eosin-Y molecule surrounded by a solvation shell given by water
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Zubkov, F I; Kvyatkovskaya, E A; Nikitina, E V; Amoyaw, P N-A; Kouznetsov, V V; Lazarenko, V A; Khrustalev, V N
2017-08-02
It has been proved that the reaction between furfuryl amines and N-R-maleimides leads to the formation of aza-Michael addition products - 3-(furylmethylamino)-N-R-pyrrolidine-2,5-diones, instead of 7-oxa-2-azabicyclo[2.2.1]hept-5-enes, as this journal reported previously.
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Synthesis and characterization of novel organotin carboxylate maleimide monomers and copolymers
Directory of Open Access Journals (Sweden)
2009-06-01
Full Text Available Two novel tributyltin carboxylate maleimide monomers, tributyltin(maleimidoacetate and tributyltin(4-maleimidobenzoate, were synthesized by condensation reaction of maleimidoacetic acid or 4-maleimidobenzoic acid with bis(tributyltin oxide. Copolymerization of these monomers with styrene was carried in dioxane at 70°C using asobisisobutyronitrile as free radical initiator. The structures of monomers and copolymers were confirmed by FT-IR (Fourier Transform Infrared, 1H and 13C NMR (nuclear magnetic resonance spectroscopy and elemental analysis. The copolymers were characterized by solubility and thermal analysis.
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Photoinduced polymerization from direct and sensitized excitation of maleimides
International Nuclear Information System (INIS)
Jonsson, S.; Lindgren, K.; Yang, D.; Viswanathan, K.; Nguyen, C.K.; Hoyle, C.
2000-01-01
Full text: The efficient photoinitiated polymerization by excited state acceptor monomers, such as maleimides (MI), has been shown in numerous acceptor/donor pair combinations. The strong hydrogen abstractability of excited triplet state MIs have also been evaluated for the initiation of acrylate polymerization and were found to be comparable in efficiency to 'the state of the art' α-cleavage type of photoinitiators. Furthermore, the importance of available abstractable hydrogens as well as the relative abstractabilities has been determined. The unique feature of MIs in general is its dual effect of simultaneously acting as a photoinitiator and as a polymerizable monomer. Once the initiation by photolysis of the acceptor / H-donors has generated a sufficient number of radicals, the unphotolyzed ground state acceptor will copolymerize with the selected monomers chosen. Although the influence of structural variations of the maleimide N-substituent, average functionality, temperature, light intensity, donor structure and acceptor / donor ratio have been carefully investigated, very little information regarding the initiating mechanism has been reported. In this paper, laser flash photolysis has been used in order to clarify and experimentally provide evidence for the mechanism of initiation. The acceptor transient intermediate generated by flash photolysis is readily quenched by typical triplet quenchers. Furthermore, selfquenching and reduction of transient lifetimes as a function of increasing concentrations of typical hydrogen donors have been determined. Stern-Volmer quenching rate constants for direct versus electron transfer induced hydrogen abstractions are calculated and compared to typical H-abstracting photoinitiators such as benzophenone (BP) and thioxanthone derivatives (ITX). From these comparisons it is clear that some triplet state MIs are more efficient in H-abstraction than the corresponding triplet states of BP and ITX. A strong and expected
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International Nuclear Information System (INIS)
Baldock, Daniel; Nocker, Andreas; Nebe-von-Caron, Gerhard; Bongaerts, Roy
2013-01-01
Unspecific background caused by biotic or abiotic particles, cellular debris, or autofluorescence is a well-known interfering parameter when applying flow cytometry to the detection of microorganisms in combination with fluorescent dyes. We present here an attempt to suppress the background signal intensity and thus to improve the detection of microorganisms using the nucleic acid stain SYBR ® Green I. It has been observed that the fluorescent signals from SYBR Green I are greatly reduced at acidic pH. When lowering the pH of pre-stained samples directly prior to flow cytometric analysis, we hypothesized that the signals from particles and cells with membrane damage might therefore be reduced. Signals from intact cells, temporarily maintaining a neutral cytosolic pH, should not be affected. We show here that this principle holds true for lowering background interference, whereas the signals of membrane-compromised dead cells are only affected weakly. Signals from intact live cells at low pH were mostly comparable to signals without acidification. Although this study was solely performed with SYBR ® Green I, the principle of low pH flow cytometry (low pH-FCM) might hold promise when analyzing complex matrices with an abundance of non-cellular matter, especially when expanded to non-DNA binding dyes with a stronger pH dependence of fluorescence than SYBR Green I and a higher pK a value. (paper)
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Wang, Li; Zhao, Hui; Chen, Yong; Sun, Ruimin; Han, Bing
2016-07-01
A high efficient homogeneous system for hydrogen production from water consisting of Eosin Y as a photosensitizer, methyl viologen (MV2+) as an electron transfer mediator, triethanolamine (TEOA) as a sacrificial electron donor and colloid Pt as a catalyst, has been systemicly studied. The initial system pH and the concentration of Eosin Y have remarkable effects on the rate of hydrogen evolution. The optimal pH and concentration of Eosin Y are 9 and 7.2×10-5 M. Triethanolamine (TEOA) as an electron donor, can reductively quench the oxidized Eosin Y and the quenching is well modeled by the Stern-Volmer equation. The optimal concentration of TEOA and the concentration of MV2+ are 0.3 M and 3.1×10-4 M, respectively. In addition, the role of colloid Pt has been investigated.
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Highly sensitive and selective fluorescent assay for guanine based on the Cu2 +/eosin Y system
Shi, Huimin; Cui, Yi; Gong, Yijun; Feng, Suling
2016-05-01
A fluorescent probe has been developed for the determination of guanine based on the quenched fluorescence signal of Cu2 +/eosin Y. Cu2 + interacted with eosin Y, resulting in fluorescence quenching. Subsequently, with the addition of guanine to the Cu2 +/eosin Y system, guanine reacted with Cu2 + to form 1:1 chelate cation, which further combined with eosin Y to form a 1:1 ternary ion-association complex by electrostatic attraction and hydrophobic interaction, resulting in significant decrease of the fluorescence. Hence, a fluorescent system was constructed for rapid, sensitive and selective detection of guanine with a detection limit as low as 1.5 nmol L- 1 and a linear range of 3.3-116 nmol L- 1. The method has been applied satisfactorily to the determination of guanine in DNA and urine samples with the recoveries from 98.7% to 105%. This study significantly expands the realm of application of ternary ion-association complex in fluorescence probe.
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Degradation of Anionic Dye Eosin by Glow Discharge Electrolysis Plasma
International Nuclear Information System (INIS)
Gao Jinzhang; Ma Dongping; Guo Xiao; Wang Aixiang; Fu Yan; Wu Jianlin; Yang Wu
2008-01-01
This paper describes a novel method for the degradation of eosin by using glow discharge electrolysis (GDE). The effects of various parameters on the removal efficiency were studied. It was found that the eosin degradation could be raised considerably by increasing the applied voltage and the initial concentration, or by decreasing pH of the aqueous solution. Fe 2+ ion had an evident accelerating effect on the eosin degradation. The degradation process of eosin obeyed a pseudo-first-order reaction. The relationship between the degradation rate constant k and the reaction temperature T could be expressed by Arrhenius equation with which the apparent activation energy Ea of 14.110 kJ. mol -1 and the pre-exponential factor ko of 2.065x10 -1 min -1 were obtained, too. The determination of hydroxyl radical was carried out by using N,N-dimethyl-p-nitrosoaniline (RNO) as a scavenger. The results showed that the hydroxyl radical plays an important role in the degradation process.
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Degradation of Anionic Dye Eosin by Glow Discharge Electrolysis Plasma
Gao, Jinzhang; Ma, Dongping; Guo, Xiao; Wang, Aixiang; Fu, Yan; Wu, Jianlin; Yang, Wu
2008-08-01
This paper describes a novel method for the degradation of eosin by using glow discharge electrolysis (GDE). The effects of various parameters on the removal efficiency were studied. It was found that the eosin degradation could be raised considerably by increasing the applied voltage and the initial concentration, or by decreasing pH of the aqueous solution. Fe2+ ion had an evident accelerating effect on the eosin degradation. The degradation process of eosin obeyed a pseudo-first-order reaction. The relationship between the degradation rate constant k and the reaction temperature T could be expressed by Arrhenius equation with which the apparent activation energy Ea of 14.110 kJ. mol-1 and the pre-exponential factor ko of 2.065×10-1 min-1 were obtained, too. The determination of hydroxyl radical was carried out by using N,N-dimethyl-p-nitrosoaniline (RNO) as a scavenger. The results showed that the hydroxyl radical plays an important role in the degradation process.
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Wilson, Thomas M.
1907-01-01
1. Eosinate of thionin gives very satisfactory staining for blood smears. It is easily prepared and dissolves readily in methylated spirits. 2. In the methods heretofore employed for making mixtures of eosin and methylene blue derivatives, the eosinates of methylene violet and methylene azure are present in very small quantities or are altogether absent. 3. Thionol and thionin are probably formed in methylene blue which has been long boiled with dilute alkalies and silver oxide. 4. Good stains of eosin and methylene blue derivatives can be obtained by a variety of manipulations. 5. Methylated spirits is more economical as a solvent for this stain and better adapted for the simple technique above described than is methyl alcohol. There is some evidence that the staining act is of a chemical nature. PMID:19867116
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Assessment of Equine Autoimmune Thrombocytopenia (EAT by flow cytometry
Directory of Open Access Journals (Sweden)
Schwarzwald Colin
2001-04-01
Full Text Available Abstract Rationale Thrombocytopenia is a platelet associated process that occurs in human and animals as result of i decreased production; ii increased utilization; iii increased destruction coupled to the presence of antibodies, within a process know as immune-mediated thrombocytopenia (IMT; or iv platelet sequestration. Thus, the differentiation of the origin of IMT and the development of reliable diagnostic approaches and methodologies are important in the clarification of IMT pathogenesis. Therefore, there is a growing need in the field for easy to perform assays for assessing platelet morphological characteristics paired with detection of platelet-bound IgG. Objectives This study is aimed to develop and characterize a single color flow cytometric assay for detection of platelet-bound IgG in horses, in combination with flow cytometric assessment of platelet morphological characteristics. Findings The FSC and SSC evaluation of the platelets obtained from the thrombocytopenic animals shows several distinctive features in comparison to the flow cytometric profile of platelets from healthy animals. The thrombocytopenic animals displayed i increased number of platelets with high FSC and high SSC, ii a significant number of those gigantic platelets had strong fluorescent signal (IgG bound, iii very small platelets or platelet derived microparticles were found significantly enhanced in one of the thrombocytopenic horses, iv significant numbers of these microplatelet/microparticles/platelet-fragments still carry very high fluorescence. Conclusions This study describes the development and characterization of an easy to perform, inexpensive, and noninvasive single color flow cytometric assay for detection of platelet-bound IgG, in combination with flow cytometric assessment of platelet morphological characteristics in horses.
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Reaction of [3H]-taurine maleimide with platelet surface thiols
International Nuclear Information System (INIS)
Karl, D.W.; Mills, D.C.B.
1986-01-01
Taurine Maleimide (2-maleimidoethanesulfonate, TM) was synthesized from [2- 3 H]-taurine and methoxycarbonylmaleimide (MCM). The yield of a 1 μmol synthesis approached 100% (based on taurine) when MCM was used in 4-fold excess. The product (TM*) was purified by ion exchange chromatography. TM* reacted irreversibly with thiol groups on the surface of washed human platelets, leading to incorporation of radioactivity into platelet pellets. Incorporation was blocked by cysteine, mercuribenzenesulfonate (MBS), dithiobisnitrobenzoate, and N-ethylmaleimide, but not by taurine or by inhibitors of anion transport. Reaction of TM* with platelets showed the dependence on time and concentration characteristics of a bimolecular reaction. The number of reactive sites ranged from 1 to 5 x 10 5 /platelet, and the apparent rate constant from 1 to 3 x 10 3 /(M x min). TM was less effective than MBS as an inhibitor of platelet aggregation induced by several agents. TM had no effect on the uptake of serotonin, taurine, or phosphate by the platelets, processes which are sensitive to MBS. These differences, considered with the similarity in size and charge of TM and MBS, suggest that classes of thiols defined as exofacial by their accessibility to MBS can differ substantially in their reactivity with other impermeant reagents
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Directory of Open Access Journals (Sweden)
Zofia Sulowska
2005-01-01
in vitro. The effect of exogenous supply of NO donors such as SNP, SIN-1, and GEA-3162 on the course of human neutrophil apoptosis and the role of extracellular antioxidants in this process was investigated. Isolated from peripheral blood, neutrophils were cultured in the presence or absence of NO donor compounds and antioxidants for 8, 12, and 20 hours. Apoptosis of neutrophils was determined in vitro by flow cytometric analysis of cellular DNA content and Annexin V protein binding to the cell surface. Exposure of human neutrophils to GEA-3162 and SIN-1 significantly accelerates and enhances their apoptosis in vitro in a time-dependent fashion. In the presence of SNP, intensification of apoptosis has not been revealed until 12 hours after the culture. The inhibition of GEA-3162- and SIN-1-mediated neutrophil apoptosis by superoxide dismutase (SOD but not by catalase (CAT was observed. Our results show that SOD and CAT can protect neutrophils against NO-donors-induced apoptosis and suggest that the interaction of NO and oxygen metabolites signals may determine the destructive or protective role of NO donor compounds during apoptotic neutrophil death.
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Luminescent Eosin Y–SiO{sub 2} hybrid nano and microrods prepared by sol–gel template method
Energy Technology Data Exchange (ETDEWEB)
Secu, M., E-mail: msecu@infim.ro [National Institute for Materials Physics, P.O. Box MG-7, Bucharest–Magurele, 077125 (Romania); Secu, C.E.; Sima, M.; Negrea, R.F.; Bartha, C. [National Institute for Materials Physics, P.O. Box MG-7, Bucharest–Magurele, 077125 (Romania); Dinescu, M.; Damian, V. [National Institute for Laser, Plasma and Radiation, P.O. Box MG-36, Bucharest–Magurele 077125 (Romania)
2013-11-15
Sol–gel chemistry within the pores of a polycarbonate template membrane was used for the preparation of Eosin Y–SiO{sub 2} hybrid nano- and microrods, using tetraethylorthosilicate [TEOS, Si(OC{sub 2}H{sub 5}){sub 4}] as the precursor in the presence of trifluoroacetic acid (TFA) catalyst. The ethanolic solution of Eosin-Y was added to the silica sol to trap dye molecules inside the SiO{sub 2} gel network during the gelation. Structural and morphological characterization using scanning electron microscopy (SEM) and luminescence microscopy have shown the formation of rods with 200 nm and 1.2 μm diameter and about 30 μm length, exhibiting luminescence properties. Spectroscopic characterization has shown that the luminescence is due to Eosin-Y molecule in the xerogel porous network, surrounded by a solvation shell given mainly by the water. -- Highlights: • Sol–gel template method was used to prepare Eosin Y–SiO{sub 2} hybrid rods-type structures. • Morphological characterization has shown nano- and microrods with luminescent properties. • Luminescence is due to Eosin-Y molecule surrounded by a solvation shell given by water.
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Huang, Zhihao; Zhao, Junfei; Wang, Zimu; Meng, Fanying; Ding, Kunshan; Pan, Xiangqiang; Zhou, Nianchen; Li, Xiaopeng; Zhang, Zhengbiao; Zhu, Xiulin
2017-10-23
Orthogonal maleimide and thiol deprotections were combined with thiol-maleimide coupling to synthesize discrete oligomers/macromolecules on a gram scale with molecular weights up to 27.4 kDa (128mer, 7.9 g) using an iterative exponential growth strategy with a degree of polymerization (DP) of 2 n -1. Using the same chemistry, a "readable" sequence-defined oligomer and a discrete cyclic topology were also created. Furthermore, uniform dendrons were fabricated using sequential growth (DP=2 n -1) or double exponential dendrimer growth approaches (DP=22n -1) with significantly accelerated growth rates. A versatile, efficient, and metal-free method for construction of discrete oligomers with tailored structures and a high growth rate would greatly facilitate research into the structure-property relationships of sophisticated polymeric materials. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.
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Highly sensitive and selective fluorescent assay for guanine based on the Cu(2+)/eosin Y system.
Shi, Huimin; Cui, Yi; Gong, Yijun; Feng, Suling
2016-05-15
A fluorescent probe has been developed for the determination of guanine based on the quenched fluorescence signal of Cu(2+)/eosin Y. Cu(2+) interacted with eosin Y, resulting in fluorescence quenching. Subsequently, with the addition of guanine to the Cu(2+)/eosin Y system, guanine reacted with Cu(2+) to form 1:1 chelate cation, which further combined with eosin Y to form a 1:1 ternary ion-association complex by electrostatic attraction and hydrophobic interaction, resulting in significant decrease of the fluorescence. Hence, a fluorescent system was constructed for rapid, sensitive and selective detection of guanine with a detection limit as low as 1.5 nmol L(-1) and a linear range of 3.3-116 nmol L(-1). The method has been applied satisfactorily to the determination of guanine in DNA and urine samples with the recoveries from 98.7% to 105%. This study significantly expands the realm of application of ternary ion-association complex in fluorescence probe. Copyright © 2016 Elsevier B.V. All rights reserved.
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Eosin Y-sensitized nanostructured SnO{sup 2}/TiO{sup 2} solar cells
Energy Technology Data Exchange (ETDEWEB)
Tai, Weon-Pil [Institute of Advanced Materials, Inha University, Yonghyun-dong, Nam-ku, Inchon 402-751 (South Korea); Inoue, Kozo [National Institute of Advanced Industrial Science and Technology, Tosu, Saga 841-0052 (Japan)
2003-02-01
The photoelectrochemical behaviors of eosin Y (organic dye)-sensitized nanostructured SnO{sub 2}/TiO{sub 2} coupled and SnO{sub 2}+TiO{sub 2} composite solar cells were studied. The value of incident photon-to-current conversion efficiency (IPCE) in the coupled system was higher than the composite system. A maximum IPCE value, 63%, was reached at 525 nm wavelength in the coupled cell with 3.5-{mu}m-thick SnO{sub 2} and 7-{mu}m-thick TiO{sub 2}. The IPCE difference in the coupled and composite cells sensitized by eosin Y dye is discussed.
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Directory of Open Access Journals (Sweden)
Jacob L Lilly
Full Text Available Targeted photopolymerization is the basis for multiple diagnostic and cell encapsulation technologies. While eosin is used in conjunction with tertiary amines as a water-soluble photoinitiation system, eosin is not widely sold as a conjugate with antibodies and other targeting biomolecules. Here we evaluate the utility of fluorescein-labeled bioconjugates to photopolymerize targeted coatings on live cells. We show that although fluorescein conjugates absorb approximately 50% less light energy than eosin in matched photopolymerization experiments using a 530 nm LED lamp, appreciable polymer thicknesses can still be formed in cell compatible environments with fluorescein photosensitization. At low photoinitiator density, eosin allows more sensitive initiation of gelation. However at higher functionalization densities, the thickness of fluorescein polymer films begins to rival that of eosin. Commercial fluorescein-conjugated antibodies are also capable of generating conformal, protective coatings on mammalian cells with similar viability and encapsulation efficiency as eosin systems.
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Lilly, Jacob L; Gottipati, Anuhya; Cahall, Calvin F; Agoub, Mohamed; Berron, Brad J
2018-01-01
Targeted photopolymerization is the basis for multiple diagnostic and cell encapsulation technologies. While eosin is used in conjunction with tertiary amines as a water-soluble photoinitiation system, eosin is not widely sold as a conjugate with antibodies and other targeting biomolecules. Here we evaluate the utility of fluorescein-labeled bioconjugates to photopolymerize targeted coatings on live cells. We show that although fluorescein conjugates absorb approximately 50% less light energy than eosin in matched photopolymerization experiments using a 530 nm LED lamp, appreciable polymer thicknesses can still be formed in cell compatible environments with fluorescein photosensitization. At low photoinitiator density, eosin allows more sensitive initiation of gelation. However at higher functionalization densities, the thickness of fluorescein polymer films begins to rival that of eosin. Commercial fluorescein-conjugated antibodies are also capable of generating conformal, protective coatings on mammalian cells with similar viability and encapsulation efficiency as eosin systems.
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Liquid-liquid extraction of uranium(VI) with cryptand-222 and Eosin as the counter ion
International Nuclear Information System (INIS)
Viji Jacob Mathew; Khopkar, S.M.
1995-01-01
Uranium(VI), (5 μg) was quantitatively extracted at pH 6.0 with 0.01M cryptand-222 in chloroform in the presence of 0.005M Eosin as the counter ion. The metal from the organic phase was stripped with 0.1M perchloric acid. Uranium(VI) from the aqueous phase was determined spectrophotometrically at 430 nm as its complex with oxine. The extraction was quantitative between pH 5.5-6.5. Nitrobenzene, chloroform and dichloromethane were the best diluents. The optimum extractant concentration was 0.01M, while that of Eosin was 0.005M. Except for perchloric acid (0.01M), other acids could not strip uranium. Uranium was separated from manganese, cadmium, lead, thallium and nickel, etc., in the multicomponent mixtures. The relative standard deviation was ±1%. (author). 18 refs., 1 fig., 6 tabs
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Directory of Open Access Journals (Sweden)
Alena S. Pankova
2016-11-01
Full Text Available 2-(Alkyl(arylaminothiazol-4(5H-ones can regioselectively be prepared from monoalkyl(arylthioureas and maleimides. In solution, the former heterocycles exist in a tautomeric equilibrium with 2-(alkyl(aryliminothiazolidin-4-ones and the substituent on the exocyclic nitrogen atom governs the ratio of these tautomers. Isomers with the alkyl group in the endocyclic position can be obtained from N-methyl(ethylthioureas. 2D NMR spectroscopy and DFT calculations rationalize experimental results.
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Ramasamy Raja, V; Rosaline, D Rani; Suganthi, A; Rajarajan, M
2018-06-01
This study presents a novel method for the preparation of Zn 2 SnO 4 /V 2 O 5 nanocomposites via a sonochemical aqueous route. This method is mild, convenient, cheap and efficient. The as prepared samples were characterized by XRD, SEM, EDAX, TEM, BET, FT-IR and UV-DRS spectra. DRS spectrum shows the adsorption edge of Zn 2 SnO 4 -V 2 O 5 in visible region of spectrum. The structural and morphological features of the as synthesized Zn 2 SnO 4 -V 2 O 5 nanocomposites have been observed using both scanning and transmission electron microscopy. BET surface area analysis inferred that the prepared hetero-junctions are meso-porous in nature. The photocatalytic activity of Zn 2 SnO 4 -V 2 O 5 nanocomposites for the degradation of Eosin Yellow (EY) dye under visible light was investigated in detail. 3% Zn 2 SnO 4 -V 2 O 5 nanocomposite exhibited the highest photocatalytic performance (92% of EY degradation) when compared with 2% Zn 2 SnO 4 -V 2 O 5 and 5% Zn 2 SnO 4 -V 2 O 5 . The adsorption of Eosin Yellow followed the pseudo-first order kinetic model. Simultaneously, high stability of the sample was also investigated by four successive photodegradation of EY under visible light. The relationship between photocatalytic activity and the structure of 3% Zn 2 SnO 4 -V 2 O 5 nanocomposite is discussed, and possible reaction mechanisms are also proposed. Therefore, the facile sonochemical preparation process provides some insight into the application of Zn 2 SnO 4 -V 2 O 5 nanocomposites in photocatalytic degradation of organic pollutants. Copyright © 2018. Published by Elsevier B.V.
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Efficacy of Eosin B as a New Antimalarial Drug in a Murine Model
Directory of Open Access Journals (Sweden)
Zahra Zamani
2012-01-01
Full Text Available The initial success of any adopted anti-infective strategy to malaria is followed by a descent due to the emergence of resistance to it. The search for new drugs and drug targets is a consistent demand in this disease. Eosin B, a common laboratory dye, is reported to have good antiparasitic properties in vitro. It was studied for its antiparasitic effect in vivo on chloroquine-sensitive Plasmodium berghei murine malaria. Eosin B was administered in 2 different doses by either the oral or parenteral route, once or twice daily to mice infected with Plasmodium berghei. Both the doses of eosin B 400 mg/kg and 800 mg/kg gave better results than the controls which were 40 mg/kg chloroquine and 100 mg/kg of arteether with P<0.005 significance. Percentage suppressive activity by Peter’s test of eosin B was better, though at a higher dose than both the controls. Survival rate of mice receiving the higher dose of eosin B was longer than that of the controls. When administered twice daily, the mice were fully cured after 4 days. Eosin B seems to be a promising drug exhibiting good antimalarial effects in the murine model of the disease.
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Investigation on the pH-dependent binding of Eosin Y and bovine serum albumin by spectral methods
International Nuclear Information System (INIS)
Gao Dejiang; Tian Yuan; Liang Fanghui; Jin Danhong; Chen Yanhua; Zhang Hanqi; Yu Aimin
2007-01-01
In this paper, the pH-dependent binding of Eosin Y and bovine serum albumin (BSA) was investigated by spectral methods, including resonance light scattering (RLS), absorption and fluorescence spectrometry. Due to the pH-dependent structure of Eosin Y and BSA, the interaction of BSA and Eosin Y depended on the solution pH value. Especially at pH 2.6 and 9.2, the RLS intensity of BSA was obviously enhanced in the presence of Eosin Y. However, the fluorescence intensity of BSA was quenched in the presence of Eosin Y. To fully understand the pH-dependent binding of BSA and Eosin Y, fluorescence quenching technique was introduced. Based on the fluorescence data obtained, the style of binding, the binding constant, the binding site number and the thermodynamic parameters for the interaction of BSA and Eosin Y were studied. Based on Foerster non-radiation energy transfer theory, the distance between donor BSA and acceptor Eosin Y was obtained
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Investigation on the pH-dependent binding of Eosin Y and bovine serum albumin by spectral methods
Energy Technology Data Exchange (ETDEWEB)
Gao Dejiang; Tian Yuan [College of Chemistry, Jilin University, Changchun 130012 (China); Liang Fanghui; Jin Danhong [Changchun Medical College, Changchun 130031 (China); Chen Yanhua; Zhang Hanqi [College of Chemistry, Jilin University, Changchun 130012 (China); Yu Aimin [College of Chemistry, Jilin University, Changchun 130012 (China)], E-mail: analchem@mail.jlu.edu.cn
2007-12-15
In this paper, the pH-dependent binding of Eosin Y and bovine serum albumin (BSA) was investigated by spectral methods, including resonance light scattering (RLS), absorption and fluorescence spectrometry. Due to the pH-dependent structure of Eosin Y and BSA, the interaction of BSA and Eosin Y depended on the solution pH value. Especially at pH 2.6 and 9.2, the RLS intensity of BSA was obviously enhanced in the presence of Eosin Y. However, the fluorescence intensity of BSA was quenched in the presence of Eosin Y. To fully understand the pH-dependent binding of BSA and Eosin Y, fluorescence quenching technique was introduced. Based on the fluorescence data obtained, the style of binding, the binding constant, the binding site number and the thermodynamic parameters for the interaction of BSA and Eosin Y were studied. Based on Foerster non-radiation energy transfer theory, the distance between donor BSA and acceptor Eosin Y was obtained.
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Radiation sensitive polymers of oxygen-substituted maleimides and elements containing same
International Nuclear Information System (INIS)
Wilson, J.C.
1977-01-01
Radiation sensitive polymers comprising from about 1 to 100 mole % of a polymerized maleimide monomer consisting of a hydrocarbon group having from 2 to 20 carbon atoms, preferably a bridged hydrocarbon group having from 6 to 10 carbon atoms and from 0 to about 99 mole % of at least one additional polymerized ethylenically unsaturated monomer are described. The polymers are soluble in organic solvents, possess desirably high glass transition temperatures and are capable of undergoing a photochemical reaction to yield polymers having isocyanate and oxy-substituted cyclopropane moieties which are capable of crosslinking in the presence of active hydrogen-containing compounds. The polymers are useful in radiation sensitive compositions and elements containing same
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[Eosin Y-water test for sperm function examination].
Zha, Shu-wei; Lü, Nian-qing; Xu, Hao-qin
2015-06-01
Based on the principles of the in vitro staining technique, hypotonic swelling test, and water test, the Eosin Y-water test method was developed to simultaneously detect the integrity of the sperm head and tail and sperm membrane structure and function. As a widely used method in clinical laboratories in China, the Eosin Y-water test is methodologically characterized by three advantages. Firstly, both the sperm head and tail can be detected at the same time, which allows easy and comprehensive assessment of membrane damage in different parts of sperm. Secondly, distilled water is used instead of the usual formula solution to simplify and standardize the test by eliminating any potential effects on the water molecules through the sperm membrane due to different osmotic pressure or different sugar proportions and electrolyte solutions. Thirdly, the test takes less time and thus can be repeated before and after treatment. This article focuses on the fundamental principles and modification of the Eosin Y-water test and its application in sperm function examination and routine semen analysis for male infertility, assessment of the quality of sperm retrieved by testicular fine needle aspiration, semen cryopreservation program development, and evaluation of sperm membrane integrity after microwave radiation.
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Eosin Removal Properties of Organo-Local Clay from Aqueous Solution
Rawan Al-Faze; Fethi Kooli
2014-01-01
Local clay from Al-Madinah Al-Munawwarah was modified by a solution of cetyltrimethylammonium bromide (C16TMABr) at different initial concentrations. The organoclays were characterized by PXRD, TGA, FTIR and N2 adsorption isotherms. The ability of these samples to remove the dye eosin is evaluated. The removal properties of organoclays were dependent on the content of C16TMA cations, the initial concentrations of eosin, temperature of the removal process, the mass of the used organoclays. T...
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Okamura, Takuma; Shimada, Yoshifumi; Nogami, Hitoshi; Kameyama, Hitoshi; Kobayashi, Takashi; Kosugi, Shin-ichi; Wakai, Toshifumi; Ajioka, Yoichi
2016-05-01
Tumor budding is recognized as an important risk factor for lymph node metastasis in pT1 colorectal cancer. Immunohistochemical staining for cytokeratin has the potential to improve the objective diagnosis of tumor budding over detection based on hematoxylin and eosin staining. However, it remains unclear whether tumor budding detected by immunohistochemical staining is a significant predictor of lymph node metastasis in pT1 colorectal cancer. The purpose of this study was to clarify the clinical significance of tumor budding detected by immunohistochemical staining in comparison with that detected by hematoxylin and eosin staining. This was a retrospective study. The study was conducted at Niigata University Medical & Dental Hospital. We enrolled 265 patients with pT1 colorectal cancer who underwent surgery with lymph node dissection. Tumor budding was evaluated by both hematoxylin and eosin and immunohistochemical staining with the use of CAM5.2 antibody. Receiver operating characteristic curve analyses were conducted to determine the optimal cutoff values for tumor budding detected by hematoxylin and eosin and CAM5.2 staining. Univariate and multivariate analyses were performed to identify the significant factors for predicting lymph node metastasis. Receiver operating characteristic curve analyses revealed that the cutoff values for tumor budding detected by hematoxylin and eosin and CAM5.2 staining for predicting lymph node metastases were 5 and 8. On multivariate analysis, histopathological differentiation (OR, 6.21; 95% CI, 1.16-33.33; p = 0.03) and tumor budding detected by hematoxylin and eosin staining (OR, 4.91; 95% CI, 1.64-14.66; p = 0.004) were significant predictors for lymph node metastasis; however, tumor budding detected by CAM5.2 staining was not a significant predictor. This study was limited by potential selection bias because surgically resected specimens were collected instead of endoscopically resected specimens. Tumor budding detected by
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Eosin fluorescence: A diagnostic tool for quantification of liver injury.
Ali, Hamid; Ali, Safdar; Mazhar, Maryam; Ali, Amjad; Jahan, Azra; Ali, Abid
2017-09-01
Hepatitis is one of the most common life threatening diseases. The diagnosis is mainly based on biochemical analysis such as liver function test. However, histopathological evaluation of liver serves far better for more accurate final diagnosis. The goal of our study was to evaluate the eosin fluorescence pattern in CCl 4 -induced liver injury model compared with normal and different treatment groups. For this purpose, liver tissues were stained with H/E and examined under bright field microscope but the fluorescence microscopy of H/E stained slides provided an interesting fluorescence pattern and was quite helpful in identifying different structures. Interesting fluorescence patterns were obtained with FITC, Texas Red and Dual channel filter cubes that were quite helpful in identifying different morphological features of the liver. During the course of hepatic injury, liver cells undergo necrosis, apoptosis and overall cellular microenvironment is altered due to the modification of proteins and other intracellular molecules. Intensified eosin fluorescence was observed around the central vein of injured liver compared to normal indicating enhanced binding of eosin to the more exposed amino acid residues. To conclude, eosin fluorescence pattern varies with the health status of a tissue and can be used further for the diagnosis and quantification of severity of various liver diseases. Copyright © 2017 Elsevier B.V. All rights reserved.
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Controlled antiseptic/eosin release from chitosan-based hydrogel modified fibrous substrates.
Romano, Ilaria; Ayadi, Farouk; Rizzello, Loris; Summa, Maria; Bertorelli, Rosalia; Pompa, Pier Paolo; Brandi, Fernando; Bayer, Ilker S; Athanassiou, Athanassia
2015-10-20
Fibers of cellulose networks were stably coated with N-methacrylate glycol chitosan (MGC) shells using subsequent steps of dip coating and photo-curing. The photo-crosslinked MGC-coated cellulose networks preserved their fibrous structure. A model hydrophilic antiseptic solution containing eosin, chloroxylenol and propylene glycol was incorporated into the shells to study the drug release dynamics. Detailed drug release mechanism into phosphate buffered saline (PBS) solutions from coated and pristine fibers loaded with the antiseptic was investigated. The results show that the MGC-coated cellulose fibers enable the controlled gradual release of the drug for four days, as opposed to fast, instantaneous release from eosin coated pristine fibers. This release behavior was found to affect the antibacterial efficiency of the fibrous cellulose sheets significantly against Staphylococcus aureus and Candida albicans. In the case of the MGC-eosin functionalized system the antibacterial efficiency was as high as 85% and 90%, respectively, while for the eosin coated pristine cellulose system the efficiency was negative, indicating bacterial proliferation. Furthermore, the MGC-eosin system was shown to be efficacious in a model of wound healing in mice, reducing the levels of various pro-inflammatory cytokines that modulate early inflammatory phase responses. The results demonstrate good potential of these coated fibers for wound dressing and healing applications. Due to its easy application on common passive commercial fibrous dressings such as gauzes and cotton fibers, the method can render them active dressings in a cost effective way. Copyright © 2015. Published by Elsevier Ltd.
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Zinc oxide based dye sensitized solar cell using eosin – Y as ...
African Journals Online (AJOL)
A zinc oxide based Dye sensitized Solar Cell (DSSC) has been fabricated, using Eosin-Y as the dye adsorbed on a nanocrystalline zinc oxide - fluorine doped tin oxide electrode, for the sensitization of the large band gap semiconductor. The absorption spectrum of Eosin-Y showed high absorption of visible light between ...
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Energy Technology Data Exchange (ETDEWEB)
Yang Qing; Zhou Ximin [National Key Laboratory of Applied Organic Chemistry, Lanzhou University, Lanzhou 730000 (China); Department of Chemistry, Lanzhou University, Lanzhou 730000 (China); Chen Xingguo, E-mail: chenxg@lzu.edu.c [National Key Laboratory of Applied Organic Chemistry, Lanzhou University, Lanzhou 730000 (China); Department of Chemistry, Lanzhou University, Lanzhou 730000 (China)
2011-04-15
The binding of Eosin B to human serum albumin (HSA) was studied using molecular docking, fluorescence, UV-vis, circular dichroism (CD) and Fourier transform infrared (FT-IR) spectroscopy. The mechanism of interaction between Eosin B and HSA in terms of the binding parameters, the thermodynamic functions and the effect of Eosin B on the conformation of HSA were investigated. Protein-ligand docking study indicated that Eosin B bound to residues located in the subdomain IIA of HSA and Eosin B-HSA complex was stabilized by hydrophobic force and hydrogen bonding. In addition, fluorescence data revealed that Eosin B strongly quenched the intrinsic fluorescence of HSA through a static quenching procedure. Furthermore, alteration of the secondary structure of HSA in the presence of the dye was conformed by UV-vis, FT-IR and CD spectroscopy.
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International Nuclear Information System (INIS)
Yang Qing; Zhou Ximin; Chen Xingguo
2011-01-01
The binding of Eosin B to human serum albumin (HSA) was studied using molecular docking, fluorescence, UV-vis, circular dichroism (CD) and Fourier transform infrared (FT-IR) spectroscopy. The mechanism of interaction between Eosin B and HSA in terms of the binding parameters, the thermodynamic functions and the effect of Eosin B on the conformation of HSA were investigated. Protein-ligand docking study indicated that Eosin B bound to residues located in the subdomain IIA of HSA and Eosin B-HSA complex was stabilized by hydrophobic force and hydrogen bonding. In addition, fluorescence data revealed that Eosin B strongly quenched the intrinsic fluorescence of HSA through a static quenching procedure. Furthermore, alteration of the secondary structure of HSA in the presence of the dye was conformed by UV-vis, FT-IR and CD spectroscopy.
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Chen, Jia-Mei; Li, Yan; Xu, Jun; Gong, Lei; Wang, Lin-Wei; Liu, Wen-Lou; Liu, Juan
2017-03-01
With the advance of digital pathology, image analysis has begun to show its advantages in information analysis of hematoxylin and eosin histopathology images. Generally, histological features in hematoxylin and eosin images are measured to evaluate tumor grade and prognosis for breast cancer. This review summarized recent works in image analysis of hematoxylin and eosin histopathology images for breast cancer prognosis. First, prognostic factors for breast cancer based on hematoxylin and eosin histopathology images were summarized. Then, usual procedures of image analysis for breast cancer prognosis were systematically reviewed, including image acquisition, image preprocessing, image detection and segmentation, and feature extraction. Finally, the prognostic value of image features and image feature-based prognostic models was evaluated. Moreover, we discussed the issues of current analysis, and some directions for future research.
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Directory of Open Access Journals (Sweden)
JAIME A. TEIXEIRA DA SILVA
2015-05-01
Full Text Available Abstract. Teixeira da Silva JA. 2015. Response of Syngonium podophyllum L. ‘White Butterfly’ shoot cultures to alternative media additives and gelling agents, and flow cytometric analysis of regenerants. Nusantara Bioscience 7: 26-32. Syngonium podophyllum L. (arrowhead vine is a popular leafy indoor pot plant whose tissue culture has been established, primarily through in vitro shoot culture, but several interesting aspects have not yet been explored. In this study, cv. ‘White Butterfly’ was used to investigate the response of shoot formation to alternative gelling agents and media additives. Gellan gum (Gelrite® at 2 g/L resulted in greater leaf production, plantlet fresh weight and higher chlorophyll content (SPAD value than all other gelling agents tested, including agar, Bacto agar, phytagel, oatmeal agar, potato dextrose agar, barley starch and corn starch, when on a basal Hyponex® (NPK = 6.5: 6: 19; 3 g/L medium. Several alternative liquid medium additives tested (low and full fat milk, Coca-Cola®, coffee, Japanese green, Oolong and Darjeeling teas negatively impacted plant growth, stunted roots and decreased chlorophyll content (SPAD value of leaves. Plant growth on medium with refined sucrose or table sugar responded similarly. Poor growth was observed when crude extract from a high rebaudioside-containing stevia (Stevia rebaudiana Bertoni line - an artificial sweetener - was used. Leaf tissue from the control did not show any endopolyploidy but low levels of endopolyploidy (8C were detected in some treatments.
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Polireddy, Kishore; Khan, Mohiuddin Md Taimur; Chavan, Hemantkumar; Young, Susan; Ma, Xiaochao; Waller, Anna; Garcia, Matthew; Perez, Dominique; Chavez, Stephanie; Strouse, Jacob J; Haynes, Mark K; Bologa, Cristian G; Oprea, Tudor I; Tegos, George P; Sklar, Larry A; Krishnamurthy, Partha
2012-01-01
ABCB6 is a member of the adenosine triphosphate (ATP)-binding cassette family of transporter proteins that is increasingly recognized as a relevant physiological and therapeutic target. Evaluation of modulators of ABCB6 activity would pave the way toward a more complete understanding of the significance of this transport process in tumor cell growth, proliferation and therapy-related drug resistance. In addition, this effort would improve our understanding of the function of ABCB6 in normal physiology with respect to heme biosynthesis, and cellular adaptation to metabolic demand and stress responses. To search for modulators of ABCB6, we developed a novel cell-based approach that, in combination with flow cytometric high-throughput screening (HTS), can be used to identify functional modulators of ABCB6. Accumulation of protoporphyrin, a fluorescent molecule, in wild-type ABCB6 expressing K562 cells, forms the basis of the HTS assay. Screening the Prestwick Chemical Library employing the HTS assay identified four compounds, benzethonium chloride, verteporfin, tomatine hydrochloride and piperlongumine, that reduced ABCB6 mediated cellular porphyrin levels. Validation of the identified compounds employing the hemin-agarose affinity chromatography and mitochondrial transport assays demonstrated that three out of the four compounds were capable of inhibiting ABCB6 mediated hemin transport into isolated mitochondria. However, only verteporfin and tomatine hydrochloride inhibited ABCB6's ability to compete with hemin as an ABCB6 substrate. This assay is therefore sensitive, robust, and suitable for automation in a high-throughput environment as demonstrated by our identification of selective functional modulators of ABCB6. Application of this assay to other libraries of synthetic compounds and natural products is expected to identify novel modulators of ABCB6 activity.
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Kovaleva, D. S.; Vikulova, M. A.; Melnikov, A. G.; Melnikov, G. V.; Gorokhovsky, A. V.
2017-03-01
The interaction of coumarin-6 and eosin-Y with particles of protonated potassium polytitanate (PPPT) was studied by luminescence spectral methods. It was established that the anisotropy of the fluorescence of coumarin-6 and eosin-Y in a suspension of PPPT is higher than in ethanol. This can be explained by bonding of the dyes with PPPT particles. The extinction coefficients of coumarin-6 (λ = 460 nm) ɛ = 25,000 M-1·cm-1 and eosin-Y (525 nm) ɛ = 36,000 M-1·cm-1 were determined from the absorption spectra of the dyes. It was shown that coumarin-6 and eosin-Y can be used for sensitization of PPPT.
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Directory of Open Access Journals (Sweden)
Qian Liu
2018-03-01
Full Text Available Selenium-containing monomer monophenyl maleimide selenide (MSM was synthesized and copolymerized with styrene (St using reversible addition-fragmentation chain transfer (RAFT polymerization. Copolymers with controlled molecular weight and narrow molecular weight distribution were obtained. The structure of the copolymer was characterized by nuclear magnetic resonance, matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF mass spectrum, Fourier transform infrared spectroscopy (FT-IR and Ultraviolet–visible spectroscopy (UV-vis spectroscopy. The copolymer can be oxidized by H2O2 to form carbon-carbon double bonds within the main chain due to the unique sensitivity of selenide groups in the presence of oxidants. Such structure changing resulted in an interesting concentration-related photoluminescence emission enhancement.
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Spychalska, Justyna; Uhrynowska, Małgorzata; Pyl, Hanna; Klimczak-Jajor, Edyta; Kopeć, Izabella; Peciakowska, Małgorzata; Gutowska, Renata; Gawlak, Maciej; Słomska, Sylwia; Dąbkowska, Syiwia; Szczecina, Roman; Dębska, Marzena; Brojer, Ewa
2015-07-01
In order to determine the appropriate dose of anti-D immunoglobulin to be administered as a preventive measure against hemolytic disease of the fetus/newborn in the subsequent pregnancy it is necessary to assess the number of fetal red blood cells that infiltrate/penetrate into the maternal circulation as a result of fetomaternal hemorrhage (FMH). One of the quantitative methods of FMH analysis is based on flow cytometry (FACS) which makes use of monoclonal antibodies to RhD antigen (anti-D test). The aim of the study was to further develop the method, evaluate its sensitivity and reproducibility and to compare it with the test based on the detection of fetal hemoglobin (HbF). The FACS study involved 20 RhD negative pregnant women and 80 RhD negative women after delivery. The following monoclonal antibodies were used: BRAD 3 FITC (anti-RhD antigen), CD45 PerCP (anti leukocyte antigen CD45), and anti-HbF PE. The fluorescence intensity of cells incubated with BRAD 3 FITC was demonstrated to depend on the RhD antigen expression, though the anti-D test also detects the weak D variant. The CD45 PerCP antibodies increased the sensitivity of anti-D test since they eliminated the leukocytes which non-specifically bind anti-D from the analysis. The presence of anti-D antibodies in maternal plasma does not affect the quantitative assessment of the fetal RhD positive fetal cells with BRAD 3 FITC. In case of FMH, the results of the anti-D test were similar to those with anti-HbF antibodies. The flow cytometric test with anti-D and anti-CD45 is useful in the assessment of the fetomaternal hemorrhage in RhD negative women. The sensitivity of the test is estimated at 0.05%.
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Photocatalytic hydrogen generation over Eosin Y-Sensitized TS-1 zeolite
International Nuclear Information System (INIS)
Zhang Xiaojie; Jin Zhiliang; Li Yuexiang; Li Shuben; Lu Gongxuan
2008-01-01
Eosin Y-sensitized TS-1 zeolite was studied for the photo-reduction of water into hydrogen driven by visible light (λ ≥ 420 nm). The optimal pH and weight ratio between Eosin Y and TS-1 zeolite is 7 and 1/8, respectively. In the presence of triethanolamine (TEA) as an electron donor, the highest rate of hydrogen generation and apparent quantum efficiency are 34 μmol h -1 and 9.4%, respectively, under visible light irradiation (λ ≥ 420 nm). Short-term stability test indicates that the catalyst is rather stable during 50 h photoreaction
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Photocatalytic hydrogen generation over Eosin Y-Sensitized TS-1 zeolite
Energy Technology Data Exchange (ETDEWEB)
Zhang Xiaojie [State Key Laboratory for Oxo Synthesis and Selective Oxidation, Lanzhou Institute of Chemical Physics, Chinese Academy of Sciences, Tianshui Zhong Road 18, Lanzhou 730000 (China); Graduate University of the Chinese Academy of Sciences, Beijing 100101 (China); Jin Zhiliang [State Key Laboratory for Oxo Synthesis and Selective Oxidation, Lanzhou Institute of Chemical Physics, Chinese Academy of Sciences, Tianshui Zhong Road 18, Lanzhou 730000 (China); Li Yuexiang [Department of Chemistry, Nanchang University, Nanjing Road 245, Nanchang 330047 (China); Li Shuben [State Key Laboratory for Oxo Synthesis and Selective Oxidation, Lanzhou Institute of Chemical Physics, Chinese Academy of Sciences, Tianshui Zhong Road 18, Lanzhou 730000 (China); Lu Gongxuan [State Key Laboratory for Oxo Synthesis and Selective Oxidation, Lanzhou Institute of Chemical Physics, Chinese Academy of Sciences, Tianshui Zhong Road 18, Lanzhou 730000 (China)], E-mail: gxlu@lzb.ac.cn
2008-05-30
Eosin Y-sensitized TS-1 zeolite was studied for the photo-reduction of water into hydrogen driven by visible light ({lambda} {>=} 420 nm). The optimal pH and weight ratio between Eosin Y and TS-1 zeolite is 7 and 1/8, respectively. In the presence of triethanolamine (TEA) as an electron donor, the highest rate of hydrogen generation and apparent quantum efficiency are 34 {mu}mol h{sup -1} and 9.4%, respectively, under visible light irradiation ({lambda} {>=} 420 nm). Short-term stability test indicates that the catalyst is rather stable during 50 h photoreaction.
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Spectral luminescence studies of eosin solvation in water-alcohol mixtures
Energy Technology Data Exchange (ETDEWEB)
Ketsle, G.A.; Levshin, L.V.; Mel' nikov, G.V.; Saletskii, A.M.
1987-11-01
The authors investigate the effects of solvation of eosin molecules in binary water-propanol mixtures with the goal of assessing eosin as a candidate dye laser material. The fluorescence was measured with a Hitachi spectrofluorimeter and the absorption spectra were taken on a Specord spectrophotometer. Absorption and fluorescence were measured for different amounts of propanol in the solvent. Data are also given on excitation and de-excitation kinetics between ground and excited states. Values for quantum yields of fluorescence and phosphorescence, average excited state lifetime, and molecular volume of the dye with the solvated shell are tabulated.
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Huisman, Andrew J.; Hartsell, Lydia R.; Krueger, Brent P.; Pikaart, Michael J.
2010-01-01
We developed a modular pair of experiments for use in the undergraduate physical chemistry and biochemistry laboratories. Both experiments examine the thermodynamics of the binding of a small molecule, eosin Y, to the protein lysozyme. The assay for binding is the quenching of lysozyme fluorescence by eosin through resonant energy transfer. In…
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Flow cytometric determination of micronucleus frequency.
Elhajouji, Azeddine; Lukamowicz-Rajska, Magdalena
2013-01-01
During the last two decades the micronucleus (MN) test has been extensively used as a genotoxicity screening tool of chemicals and in a variety of exploratory and mechanistic investigations. The MN is a biomarker for chromosomal damage or mitotic abnormalities, since it can originate from chromosome fragments or whole chromosomes that fail to be incorporated into daughter nuclei during mitosis (Fenech et al., Mutagenesis 26:125-132, 2011; Kirsch-Volders et al., Arch Toxicol 85:873-899, 2011). The simplicity of scoring, accuracy, amenability to automation by image analysis or flow cytometry, and readiness to be applied to a variety of cell types either in vitro or in vivo have made it a versatile tool that has contributed to a large extent in our understanding of key toxicological issues related to genotoxins and their effects at the cellular and organism levels. Recently, the final acceptance of the in vitro MN test guideline 487 (OECD Guideline for Testing of Chemicals, In vitro mammalian cell micronucleus test 487. In vitro mammalian cell micronucleus test (MNVIT). Organization for Economic Cooperation and Development, Paris, 2010) together with the standard in vivo MN test OECD guideline 474 (OECD Guideline for The Testing of Chemicals, Mammalian erythrocyte micronucleus test no. 474. Organization for Economic Cooperation and Development, Paris, 1997) will further position the assay as a key driver in the determination of the genotoxicity potential in exploratory research as well as in the regulatory environment. This chapter covers to some extent the protocol designs and experimental steps necessary for a successful performance of the MN test and an accurate analysis of the MN by the flow cytometry technique.
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Rubak, Peter; Kristensen, Steen D; Hvas, Anne-Mette
2017-06-01
Immature platelets may contain more platelet enzymes such as cyclooxygenase (COX)-1 and COX-2 than mature platelets. Patients with immune thrombocytopenia (ITP) have a higher fraction of immature platelets and can therefore be utilized as a biological model for investigating COX-1 and COX-2 platelet expression. The aims were to develop flow cytometric assays for platelet COX-1 and COX-2 and to investigate the COX-1 and COX-2 platelet expression, platelet turnover, and platelet glycoproteins in ITP patients (n = 10) compared with healthy individuals (n = 30). Platelet count and platelet turnover parameters (mean platelet volume (MPV), immature platelet fraction (IPF), and immature platelet count (IPC)) were measured by flow cytometry (Sysmex XE-5000). Platelet COX-1, COX-2, and the glycoproteins (GP)IIb, IX, Ib, Ia, and IIIa were all analyzed by flow cytometry (Navios) and expressed as median fluorescence intensity. COX analyses were performed in both whole blood and platelet rich plasma (PRP), whereas platelet glycoproteins were analyzed in whole blood only. ITP patients had significantly lower platelet count (55 × 10 9 /L) than healthy individuals (240 × 10 9 /L, p platelet count and IPC (both p-values Platelet COX-1 expression was higher in ITP patients than healthy individuals using whole blood (p COX-1 platelet turnover and COX-1 expression (all p-values platelet turnover and COX-1 and COX-2 expressions (all p-values platelet turnover in ITP patients (all p-values 0.14, rho = 0.11-0.28). In conclusion, ITP patients expressed higher COX-1 and platelet glycoprotein levels than healthy individuals. COX-1 and platelet glycoproteins demonstrated positive correlations with platelet turnover in ITP patients. In healthy individuals, COX-1 and COX-2 expression correlated positively with platelet turnover. PRP was more sensitive compared with whole blood as regards determination of COX. Therefore, PRP is the recommended matrix for investigating COX-1 and COX-2 in
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Directory of Open Access Journals (Sweden)
Daniel W. Visscher
1998-01-01
Full Text Available Two-color, multiparametric synthesis phase fraction (SPF analysis of cytokeratin-labeled epithelial cells was flow cytometrically performed on both benign (SPFb and malignant tissue samples (if available, SPFt from 132 mastectomy/lumpectomy specimens. These data were then correlated with clinicopathologic features, including (1 tumor differentiation, (2 the proportion of tumor comprised of duct carcinoma-in situ (DCIS, and (3 the histology of accompanying benign breast tissue, classified by predominant microscopic pattern as intact, normal terminal duct lobular units (NTDLU, 34% of cases, atrophic (AT, 33% of cases, proliferative fibrocystic (PFC, 26% of cases, and non-proliferative fibrocystic (NPFC, 7% of cases. SPFt was inversely correlated with extent of DCIS (DCIS =0 – 20% tumor volume – 12.7% mean SPFt, vs. DCIS >20% tumor volume – 6.4% mean SPFt, p = 0.001. SPFt also correlated with the histology of background benign breast tissue (NTDLU – 14.8% mean SPFt vs. AT – 6.9% mean SPFt vs. PFC – 12.7% mean SPFt, p = 0.05 but it did not correlate with patient age or SPFb (overall mean =0.73%. SPFb was correlated with patient age (>56 yr – 0.59% mean SPFb vs. < yr – 0.84% mean SPFb, p = 0.02, with background histology (NTDLU – 1.1% mean SPFb vs. AT – 0.43% mean SPFb vs. PFC – 0.70% mean SPFb, p < 0.02 and with the grade of the neoplasm (well/moderate – 0.58% mean vs. poorly differentiated – 0.85% mean, p = 0.04. Patients having a background of PFC were significantly older than patients with a background of NTDLU (45.2 yr vs. 60.2 yr, p = 0.01.
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Directory of Open Access Journals (Sweden)
Kishore Polireddy
Full Text Available ABCB6 is a member of the adenosine triphosphate (ATP-binding cassette family of transporter proteins that is increasingly recognized as a relevant physiological and therapeutic target. Evaluation of modulators of ABCB6 activity would pave the way toward a more complete understanding of the significance of this transport process in tumor cell growth, proliferation and therapy-related drug resistance. In addition, this effort would improve our understanding of the function of ABCB6 in normal physiology with respect to heme biosynthesis, and cellular adaptation to metabolic demand and stress responses. To search for modulators of ABCB6, we developed a novel cell-based approach that, in combination with flow cytometric high-throughput screening (HTS, can be used to identify functional modulators of ABCB6. Accumulation of protoporphyrin, a fluorescent molecule, in wild-type ABCB6 expressing K562 cells, forms the basis of the HTS assay. Screening the Prestwick Chemical Library employing the HTS assay identified four compounds, benzethonium chloride, verteporfin, tomatine hydrochloride and piperlongumine, that reduced ABCB6 mediated cellular porphyrin levels. Validation of the identified compounds employing the hemin-agarose affinity chromatography and mitochondrial transport assays demonstrated that three out of the four compounds were capable of inhibiting ABCB6 mediated hemin transport into isolated mitochondria. However, only verteporfin and tomatine hydrochloride inhibited ABCB6's ability to compete with hemin as an ABCB6 substrate. This assay is therefore sensitive, robust, and suitable for automation in a high-throughput environment as demonstrated by our identification of selective functional modulators of ABCB6. Application of this assay to other libraries of synthetic compounds and natural products is expected to identify novel modulators of ABCB6 activity.
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International Nuclear Information System (INIS)
Du Wenli; Xu Zirong; Han Xinyan; Xu Yinglei; Miao Zhiguo
2008-01-01
The present study dealt with the adsorption of eosin Y, as a model anionic dye, from aqueous solution using chitosan nanoparticles prepared by the ionic gelation between chitosan and tripolyphosphate. The nanoparticles were characterized by atomic force microscopy (AFM), size and zeta potential analysis. A batch system was applied to study the adsorption of eosin Y from aqueous solution by chitosan nanoparticles. The results showed that the adsorption of eosin Y on chitosan nanoparticles was affected by contact time, eosin Y concentration, pH and temperature. Experimental data followed Langmuir isotherm model and the adsorption capacity was found to be 3.333 g/g. The adsorption process was endothermic in nature with an enthalpy change (ΔH) of 16.7 kJ/mol at 20-50 deg. C. The optimum pH value for eosin Y removal was found to be 2-6. The dye was desorbed from the chitosan nanoparticles by increasing the pH of the solution
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Energy Technology Data Exchange (ETDEWEB)
Du Wenli [Institute of Feed Science, College of Animal Science, Zhejiang University, Key Laboratory of Molecular Animal Nutrition, Ministry of Education, Hangzhou 310029 (China)], E-mail: wenlidu@126.com; Xu Zirong; Han Xinyan; Xu Yinglei; Miao Zhiguo [Institute of Feed Science, College of Animal Science, Zhejiang University, Key Laboratory of Molecular Animal Nutrition, Ministry of Education, Hangzhou 310029 (China)
2008-05-01
The present study dealt with the adsorption of eosin Y, as a model anionic dye, from aqueous solution using chitosan nanoparticles prepared by the ionic gelation between chitosan and tripolyphosphate. The nanoparticles were characterized by atomic force microscopy (AFM), size and zeta potential analysis. A batch system was applied to study the adsorption of eosin Y from aqueous solution by chitosan nanoparticles. The results showed that the adsorption of eosin Y on chitosan nanoparticles was affected by contact time, eosin Y concentration, pH and temperature. Experimental data followed Langmuir isotherm model and the adsorption capacity was found to be 3.333 g/g. The adsorption process was endothermic in nature with an enthalpy change ({delta}H) of 16.7 kJ/mol at 20-50 deg. C. The optimum pH value for eosin Y removal was found to be 2-6. The dye was desorbed from the chitosan nanoparticles by increasing the pH of the solution.
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Energy Technology Data Exchange (ETDEWEB)
Santos, N.F.G.; Amaral, A., E-mail: neyliane@gmail.com [Universidade Federal de Pernambuco (UFPE), Recife, PE (Brazil). Departamento de Energia Nuclear. Laboratorio de Modelagem e Biodosimetria Aplicada; Freitas-Silva, R. [Universidade Federal de Pernambuco (UFPE), Garanhuns, PE (Brazil). Departamento de Ciencias Naturais e Exatas; Pereira, V.R.A. [Fundacao Oswaldo Cruz (FIOCRUZ), Recife, PE (Brazil). Centro de Pesquisas Aggeu Magalhaes. Departamento de Imunologia. Lab. de Imunoparasitologia; Tasat, D.R. [Universidad Nacional de General San Martin, Buenos Aires (Argentina). Escuela de Ciencia y Tecnologia. Laboratorio de Biologia Celular del Pulmon
2013-08-15
Cell cycle blockage in G1 is a mechanism p21 protein-regulated and coupled to DNA damage response to permit genetic content analysis, damage repair and cell death. Analysis of proteins that participates of this response has progressed with new analytic tools, and data contributes to comprehension of radioinduced molecular events as well as to new approaches on practices that employ ionizing radiation. On this perspective, the aim of this research was to evaluate, by flow cytometry, p21 expression on irradiated human lymphocytes, maintained under different experimental conditions. Peripheral blood samples from 10 healthy subjects were irradiated with doses of 0 (non-irradiated), 1, 2 and 4 Gy. Lymphocytes were processed to analysis on ex vivo (no cultured) condition and after 24; 48 and 72 hours culture, with and without phytohemagglutinin stimulation. p21 protein expression levels were measured by flow cytometry, as percentage values. Results indicate that flow cytometric assay allows detection of changes on p21 expression, since it was detected significant increase on phytohemagglutinin-stimulated samples, for all times, against basal expression (ex vivo). However, it was not observed significant alterations on p21 protein radioinduced levels, for all doses, times and culture conditions analyzed. These results not indicate so p21 protein as bioindicator of ionizing radiation exposure. Nevertheless, data confirmation may to require analysis of a more numerous population. (author)
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Influence of the supporting salt concentration on the electrodeposition of ZnO/eosin Y hybrid films
Energy Technology Data Exchange (ETDEWEB)
Boeckler, C.; Oekermann, T. [Institute of Physical Chemistry and Electrochemistry, Leibniz University Hannover (Germany); Saruban, M.; Ichinose, K.; Yoshida, T. [Center of Innovative Photovoltaic Systems (CIPS), Environmental and Renewable Energy Systems (ERES) Division, Graduate School of Engineering, Gifu University (Japan)
2008-10-15
ZnO/eosin Y hybrid films were electrodeposited from O{sub 2}-saturated ZnCl{sub 2}-solution under variation of the concentration of the supporting salt KCl. The supporting salt concentration was found to have a significant influence on the amount of dye loading and the morphology of the ZnO/eosin Y films. By decreasing the supporting salt concentration to 0.01 M KCl, porous films with a high dye loading can be obtained at potentials of -0.8 V vs. SCE at low eosin Y concentrations in the electrodeposition bath, which usually only lead to porous films at potentials where eosin Y is reduced (<-0.9 V vs. SCE). (copyright 2008 WILEY-VCH Verlag GmbH and Co. KGaA, Weinheim) (orig.)
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Electron microscopy of rapid identification of animal viruses in hematoxylin-eosin sections.
Bhatnagar, R; Johnson, G R; Christian, R G
1977-01-01
Routine hematoxylin-eosin stained, paraffin sections were processed for electron microscopy, using a rapid method for localization of animal viruses. Formalin fixation was effective in preserving DNA as well as RNA viruses, however cellular fine structural details and organelles were not well preserved. The procedure is useful for morphological recognition of viral groups and as a rapid diagnostic aid for identifying viral disease. Images Fig. 1. Fig. 2. Fig. 3. Fig. 4. Fig. 5. Fig. 6. PMID:72592
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Solid KHT tumor dispersal for flow cytometric cell kinetic analysis
International Nuclear Information System (INIS)
Pallavicini, M.G.; Folstad, L.J.; Dunbar, C.
1981-01-01
A bacterial neutral protease was used to disperse KHT solid tumors into single cell suspensions suitable for routine cell kinetic analysis by flow cytometry and for clonogenic cell survival. Neutral protease disaggregation under conditions which would be suitable for routine tumor dispersal was compared with a trypsin/DNase procedure. Cell yield, clonogenic cell survival, DNA distributions of untreated and drug-perturbed tumors, rates of radioactive precursor incorporation during the cell cycle, and preferential cell cycle phase-specific cell loss were investigated. Tumors dispersed with neutral protease yielded approximately four times more cells than those dispersed with trypsin/DNase and approximately a 1.5-fold higher plating efficiency in a semisolid agar system. Quantitative analysis of DNA distributions obtained from untreated and cytosine-arabinoside-perturbed tumors produced similar results with both dispersal procedures. The rates of incorporation of tritiated thymidine during the cell cycle were also similar with neutral protease and trypsin/DNase dispersal. Preferential phase-specific cell loss was not obseved with either technique. We find that neutral protease provides good single cell suspensions of the KHT tumor for cell survival measurements and for cell kinetic analysis of drug-induced perturbations by flow cytometry. In addition, the high cell yields facilitate electronic cell sorting where large numbers of cells are often required
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Directory of Open Access Journals (Sweden)
Amandeep Kaur
2012-01-01
Full Text Available H-point standard addition method (HPSAM is developed for simultaneous determination of eosin-Y and ponceau-s in micellar media. Nickel chloride (NiCl2 is used as chromogenic reagent for complexes formation of eosin-Y and ponceau-S food colorants. The measurements were carried out using sodium lauryl sulphate as a surfactant, in buffered solution at pH 6.0. The concentration range of 0.115-2.53 μg/mL of eosin-Y and 0.159-3.80 μg/mL of ponceau-S. The proposed procedures have been applied successfully for the simultaneous determination of eosin-Y and ponceau-S in synthetic binary mixtures and real samples.
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Physiological pH fiber-optic chemical sensor based on energy transfer. [Eosin and phenol red
Energy Technology Data Exchange (ETDEWEB)
Jordan, D.M.; Walt, D.R.; Milanovich, F.P.
1987-02-01
A fiber-optic sensor has been developed containing a fluorophore, eosin, and an absorber, phenol red, coimmobilized on the distal end of an optical fiber. When an argon laser is used to excite eosin with light of lambda 488 nm, a region of the spectrum where phenol red does not absorb, eosin emits light in a spectral region that overlaps significantly with the absorption spectru of the basic form of phenol red. Consequently, nonradiative energy transfer occurs from eosin (donor) to phenol red (acceptor). The amount of energy transfer increases as the pH increases resulting in a diminished fluorescence intensity. Thus, changes in the absorption of phenol red as a function of pH are detected as changes in the fluorescent signal. In this manner a pH sensor optimized for physiological pH measurement has been prepared. The fiber exhibits a precision of at least 0.01 pH units.
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Van Der Merwe, Riaan
2014-06-01
The discharge of concentrate and other process waters from seawater reverse osmosis (SWRO) plant operations into the marine environment may adversely affect water quality in the near-field area surrounding the outfall. The main concerns are the increase in salt concentration in receiving waters, which results in a density increase and potential water stratification near the outfall, and possible increases in turbidity, e.g., due to the discharge of filter backwash waters. Changes in ambient water quality may affect microbial abundance in the area, for example by hindering the photosynthesis process or disrupting biogenesis. It is widely accepted that marine biodiversity is lower in more extreme conditions, such as high salinity environments. As aquatic microbial communities respond very rapidly to changes in their environment, they can be used as indicators for monitoring ambient water quality. The objective of this study was to assess possible changes in microbial abundance as a result of concentrate discharge into the near-field area (<. 25. m) surrounding the outfall of the King Abdullah University of Science and Technology (KAUST) SWRO plant. Flow cytometric (FCM) analysis was conducted in order to rapidly determine microbial abundance on a single-cell level in 107 samples, taken by diving, from the discharge area, the intake area and two control sites. FCM analysis combined the measurement of distinct scatter of cells and particles, autofluorescence of cyanobacteria and algae, and fluorescence after staining of nucleic acids with SYBR® Green for a total bacterial count. The results indicate that changes in microbial abundance in the near-field area of the KAUST SWRO outfall are minor and appear to be the result of a dilution effect rather than a direct impact of the concentrate discharge. © 2014 Elsevier B.V.
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Intermolecular Interactions between Eosin Y and Caffeine Using 1H-NMR Spectroscopy
Directory of Open Access Journals (Sweden)
Macduff O. Okuom
2013-01-01
Full Text Available DETECHIP has been used in testing analytes including caffeine, cocaine, and tetrahydrocannabinol (THC from marijuana, as well as date rape and club drugs such as flunitrazepam, gamma-hydroxybutyric acid (GHB, and methamphetamine. This study investigates the intermolecular interaction between DETECHIP sensor eosin Y (DC1 and the analyte (caffeine that is responsible for the fluorescence and color changes observed in the actual array. Using 1H-NMR, 1H-COSY, and 1H-DOSY NMR methods, a proton exchange from C-8 of caffeine to eosin Y is proposed.
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Ni, Yongnian; Liu, Qiuhong; Kokot, Serge
2011-01-01
Interaction of chlorotetracycline (CTC) with bovine serum albumin (BSA) was investigated under simulated physiological conditions by spectroscopy with the aid of multivariate curve resolution-alternating least squares (MCR-ALS). Eosin Y was selected as an alternative site I marker on the BSA to study the above molecular interaction. The binding of Eosin Y and CTC to BSA showed that CTC was displaced from CTC-BSA complex by Eosin Y, and Eosin Y-BSA complex was formed. However, the recorded fluorescence spectra of Eosin Y and Eosin Y-BSA overlapped and MCR-ALS was applied to resolve the two-way fluorescence spectra. From the resolved equilibrium concentration profiles, it was observed that Eosin Y competed with CTC in the binding process with BSA; it was also shown that the binding site of CTC on BSA was site I, and this was further confirmed by the fluorescence polarization method. Compared with some common site I markers for BSA, the fluorescence and UV-vis spectral shapes of the Eosin Y-BSA complex were quite different from that of Eosin Y, and this feature facilitated the investigation of the small molecule-BSA interaction.
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Designed plasmonic nanocatalysts for the reduction of eosin Y: absorption and fluorescence study
Komalam, Abha; Muraleegharan, Lekha Girija; Subburaj, Suganthi; Suseela, Suji; Babu, Aswathy; George, Sony
2012-10-01
In this work, we report a one-step green synthesis of gold nanoparticles (AuNPs) by microwave irradiation using nontoxic and biodegradable polysaccharide chitosan as a reducing and stabilizing agent. The interaction between gold nanoparticles with the amine group of chitosan was confirmed by Fourier transform infrared spectroscopy analysis, and the stability of the nanoparticle is ascertained by zeta potential measurements. Transmission electron microscopy photograph and dynamic light scattering measurements confirmed the average size of gold nanoparticles as 25 nm. The ability of the synthesised gold nanoparticles as a catalyst for the reduction of eosin dye in the presence of NaBH4 was monitored by means of spectrofluorometry and spectrophotometry. It is found that the NaBH4-induced reduction of eosin is enhanced in the presence of AuNPs even without a catalyst. Time-resolved fluorescence decay studies also confirmed the reduction of eosin in the presence of AuNPs.
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Bellal, B.; Trari, M.; Afalfiz, A.
2015-08-01
The advantages of the hetero-junction CdS/CuAl2O4 for the photocatalytic eosin degradation are reported. Composite semiconductors are elaborated by co-precipitation of CdS on the spinel CuAl2O4 giving a core-shell structure with a uniform dispersion and intimate contact of the spinel nanoparticles inside the hexagonal CdS. The Mott-Schottky plots ( C -2- V) of both materials show linear behaviors from which flat band potentials are determined. The photoactivity increases with increasing the mass of the sensitizer CdS and the best performance is achieved on CdS/CuAl2O4 (85 %/15 %). The pH has a strong influence on the degradation and the photoactivity peaks at pH 7.78. The dark adsorption eosin is weak (~4 %), hence the change in the eosin concentration is attributed to the photocatalytic process. The degradation follows a zero-order kinetic with a rate constant of 5.2 × 10-8 mol L-1 mn-1 while that of the photolysis is seven times lower (0.75 × 10-8 mol L-1 mn-1).
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Directory of Open Access Journals (Sweden)
Seong-Soo Roh
2008-01-01
Full Text Available The present study is performed to investigate the inhibitory effects of Radix Adenophorae extract (RAE on ovalbumin-induced asthma murine model. To study the anti-inflammatory and antiasthmatic effects of RAE, we examined the development of pulmonary eosinophilic inflammation and inhibitory effects of T cells in murine by RAE and cyclosporine A (CsA. We examined determination of airway hyperresponsiveness, flow cytometric analysis (FACS, enzyme-linked immunosorbent assay (ELISA, quantitative real time (PCR, hematoxylin-eosin staining, and Masson trichrome staining in lung tissue, lung weight, total cells, and eosinophil numbers in lung tissue. We demonstrated how RAE suppressed development on inflammation and decreased airway damage.
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Bradford, Jolene A; Buller, Gayle; Suter, Michael; Ignatius, Michael; Beechem, Joseph M
2004-10-01
Conventional immuno-based multiparameter flow cytometric analysis has been limited by the requirement of a dedicated detection channel for each antibody-fluorophore set. To address the need to resolve multiple biological targets simultaneously, flow cytometers with as many as 10-15 detection channels have been developed. In this study, a new Zenon immunolabeling technology is developed that allows for multiple antigen detection per detection channel using a single fluorophore, through a unique method of fluorescence-intensity multiplexing. By varying the Zenon labeling reagent-to-antibody molar ratio, the fluorescence intensity of the antibody-labeled cellular targets can be used as a unique identifier. Although demonstrated in the present study with lymphocyte immunophenotyping, this approach is broadly applicable for any immuno-based multiplexed flow cytomety assay. Lymphocyte immunophenotyping of 38 clinical blood specimens using CD3, CD4, CD8, CD16, CD56, CD19, and CD20 antibodies was performed using conventional flow cytometric analysis and fluorescence-intensity multiplexing analysis. Conventional analysis measures a single antibody-fluorophore per photomultiplier tube (PMT). Fluorescence-intensity multiplex analysis simultaneously measures seven markers with two PMTs, using Zenon labeling reagent-antibody complexes in a single tube: CD19, CD4, CD8, and CD16 antibodies labeled with Zenon Alexa Fluor 488 Mouse IgG(1) labeling reagent and CD56, CD3, and CD20 antibodies labeled with Zenon R-Phycoerythrin (R-PE) Mouse IgG(1) or IgG(2b) labeling reagents. The lymphocyte immunophenotyping results from fluorescence-intensity multiplexing using Zenon labeling reagents in a single tube were comparable to results from conventional flow cytometric analysis. Simultaneous evaluation of multiple antigens using a single fluorophore can be performed using antibodies labeled with varying ratios of a Zenon labeling reagent. Labeling two sets of antibodies with different Zenon
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Directory of Open Access Journals (Sweden)
Gregory A Johnson
Full Text Available BACKGROUND/OBJECTIVES: Upon irradiation with visible light, the photosensitizer-peptide conjugate eosin-(KLAKLAK2 kills a broad spectrum of bacteria without damaging human cells. Eosin-(KLAKLAK2 therefore represents an interesting lead compound for the treatment of local infection by photodynamic bacterial inactivation. The mechanisms of cellular killing by eosin-(KLAKLAK2, however, remain unclear and this lack of knowledge hampers the development of optimized therapeutic agents. Herein, we investigate the localization of eosin-(KLAKLAK2 in bacteria prior to light treatment and examine the molecular basis for the photodynamic activity of this conjugate. METHODOLOGY/PRINCIPAL FINDINGS: By employing photooxidation of 3,3-diaminobenzidine (DAB, (scanning transmission electron microscopy ((STEM, and energy dispersive X-ray spectroscopy (EDS methodologies, eosin-(KLAKLAK2 is visualized at the surface of E. coli and S. aureus prior to photodynamic irradiation. Subsequent irradiation leads to severe membrane damage. Consistent with these observations, eosin-(KLAKLAK2 binds to liposomes of bacterial lipid composition and causes liposomal leakage upon irradiation. The eosin moiety of the conjugate mediates bacterial killing and lipid bilayer leakage by generating the reactive oxygen species singlet oxygen and superoxide. In contrast, the (KLAKLAK2 moiety targets the photosensitizer to bacterial lipid bilayers. In addition, while (KLAKLAK2 does not disrupt intact liposomes, the peptide accelerates the leakage of photo-oxidized liposomes. CONCLUSIONS/SIGNIFICANCE: Together, our results suggest that (KLAKLAK2 promotes the binding of eosin Y to bacteria cell walls and lipid bilayers. Subsequent light irradiation results in membrane damage from the production of both Type I & II photodynamic products. Membrane damage by oxidation is then further aggravated by the (KLAKLAK2 moiety and membrane lysis is accelerated by the peptide. These results therefore
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Image cytometric nuclear texture features in inoperable head and neck cancer: a pilot study
International Nuclear Information System (INIS)
Strojan-Flezar, Margareta; Lavrencak, Jaka; Zganec, Mario; Strojan, Primoz
2011-01-01
Image cytometry can measure numerous nuclear features which could be considered a surrogate end-point marker of molecular genetic changes in a nucleus. The aim of the study was to analyze image cytometric nuclear features in paired samples of primary tumor and neck metastasis in patients with inoperable carcinoma of the head and neck. Image cytometric analysis of cell suspensions prepared from primary tumor tissue and fine needle aspiration biopsy cell samples of neck metastases from 21 patients treated with concomitant radiochemotherapy was performed. Nuclear features were correlated with clinical characteristics and response to therapy. Manifestation of distant metastases and new primaries was associated (p<0.05) with several chromatin characteristics from primary tumor cells, whereas the origin of index cancer and disease response in the neck was related to those in the cells from metastases. Many nuclear features of primary tumors and metastases correlated with the TNM stage. A specific pattern of correlation between well-established prognostic indicators and nuclear features of samples from primary tumors and those from neck metastases was observed. Image cytometric nuclear features represent a promising candidate marker for recognition of biologically different tumor subgroups
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Basuli, Falguni; Zhang, Xiang; Jagoda, Elaine M; Choyke, Peter L; Swenson, Rolf E
2018-03-25
Following our recently published fluorine-18 labeling method, "Radio-fluorination on the Sep-Pak", we have successfully synthesized 6-[ 18 F]fluoronicotinaldehyde by passing a solution (1:4 acetonitrile: t-butanol) of its quaternary ammonium salt precursor, 6-(N,N,N-trimethylamino)nicotinaldehyde trifluoromethanesulfonate (2), through a fluorine-18 containing anion exchange cartridge (PS-HCO 3 ). Over 80% radiochemical conversion was observed using 10 mg of precursor within 1 minute. The [ 18 F]fluoronicotinaldehyde ([ 18 F]5) was then conjugated with 1-(6-(aminooxy)hexyl)-1H-pyrrole-2,5-dione to prepare the fluorine-18 labeled maleimide functionalized prosthetic group, 6-[ 18 F]fluoronicotinaldehyde O-(6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexyl) oxime, 6-[ 18 F]FPyMHO ([ 18 F]6). The current Sep-Pak method not only improves the overall radiochemical yield (50 ± 9%, decay-corrected, n = 9) but also significantly reduces the synthesis time (from 60-90 minutes to 30 minutes) when compared with literature methods for the synthesis of similar prosthetic groups. Published 2018. This article is a U.S. Government work and is in the public domain in the USA.
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Lyoluminescence studies using Eosin B as a sensitizer
International Nuclear Information System (INIS)
Galand, E.; Niezette, J.; Vanderschueren, J.; Vanderschueren, H.W.
1991-01-01
Lyoluminescence of various saccharides and of glutamine has been investigated by using a home made read-out system allowing a better control of the dissolution rate. The device has been tested by measuring the weight and dose dependence of D(+) mannose and the influence of the origin of the sample. The use of eosin B solutions as a solvent shows a substantial enhancement of lyoluminescence for dextrose and xylose. (author)
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Lyoluminescence studies using Eosin B as a sensitizer
Energy Technology Data Exchange (ETDEWEB)
Galand, E.; Niezette, J.; Vanderschueren, J. (Inst. de Chimie, Sart Tilman, Liege (Belgium)); Vanderschueren, H.W. (Inst. d' Electrotechnique Montefiore, Sart Tilman, Liege (Belgium))
1991-01-01
Lyoluminescence of various saccharides and of glutamine has been investigated by using a home made read-out system allowing a better control of the dissolution rate. The device has been tested by measuring the weight and dose dependence of D(+) mannose and the influence of the origin of the sample. The use of eosin B solutions as a solvent shows a substantial enhancement of lyoluminescence for dextrose and xylose. (author).
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Vignesh, K.; Suganthi, A.; Rajarajan, M.; Sakthivadivel, R.
2012-03-01
Hesperidin a flavanoid, modified TiO2 nanoparticles (Hes-TiO2) was synthesized to improve the visible light driven photocatalytic performance of TiO2. The synthesized nanoparticles were characterized by UV-visible diffuse reflectance spectroscopy (UV-vis-DRS), FT-IR, powder X-ray diffraction (XRD) and scanning electron microscopy (SEM). The photocatalytic activity of Hes-TiO2 was investigated based on the decolorization of eosin-Y under visible light irradiation. Hes-TiO2 showed high efficiency for the decolorization of eosin-Y. The influences of various reaction parameters like effect of pH, catalyst dosage and initial dye concentration on the photocatalytic efficiency were investigated. The adsorption of eosin-Y on Hes-TiO2 was found favorable by the Langmuir approach. The removal percentage of chemical oxygen demand (COD) was determined to evaluate the mineralization of eosin-Y during photodecolorization. Based on the intermediates obtained in the GC-MS spectroscopic technique, a probable degradation mechanism has been proposed.
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International Nuclear Information System (INIS)
Vignesh, K.; Suganthi, A.; Rajarajan, M.; Sakthivadivel, R.
2012-01-01
Hesperidin a flavanoid, modified TiO 2 nanoparticles (Hes-TiO 2 ) was synthesized to improve the visible light driven photocatalytic performance of TiO 2 . The synthesized nanoparticles were characterized by UV-visible diffuse reflectance spectroscopy (UV-vis-DRS), FT-IR, powder X-ray diffraction (XRD) and scanning electron microscopy (SEM). The photocatalytic activity of Hes-TiO 2 was investigated based on the decolorization of eosin-Y under visible light irradiation. Hes-TiO 2 showed high efficiency for the decolorization of eosin-Y. The influences of various reaction parameters like effect of pH, catalyst dosage and initial dye concentration on the photocatalytic efficiency were investigated. The adsorption of eosin-Y on Hes-TiO 2 was found favorable by the Langmuir approach. The removal percentage of chemical oxygen demand (COD) was determined to evaluate the mineralization of eosin-Y during photodecolorization. Based on the intermediates obtained in the GC-MS spectroscopic technique, a probable degradation mechanism has been proposed.
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Energy Technology Data Exchange (ETDEWEB)
Vanparys, Caroline, E-mail: caroline.vanparys@ua.ac.be [Laboratory of Ecophysiology, Biochemistry and Toxicology, University of Antwerp, Antwerp (Belgium); Depiereux, Sophie; Nadzialek, Stephanie [Research Unit in Organismal Biology (URBO), University of Namur (FUNDP), Namur (Belgium); Robbens, Johan; Blust, Ronny [Laboratory of Ecophysiology, Biochemistry and Toxicology, University of Antwerp, Antwerp (Belgium); Kestemont, Patrick [Research Unit in Organismal Biology (URBO), University of Namur (FUNDP), Namur (Belgium); De Coen, Wim [Laboratory of Ecophysiology, Biochemistry and Toxicology, University of Antwerp, Antwerp (Belgium); European Chemicals Agency (ECHA), Helsinki (Finland)
2010-09-15
In vitro estrogenicity screens are believed to provide a first prioritization step in hazard characterization of endocrine disrupting chemicals. When applied to complex environmental matrices or mixture samples, they have been indicated valuable in estimating the overall estrogen-mimicking load. In this study, the performance of an adapted format of the classical E-screen or MCF-7 cell proliferation assay was profoundly evaluated to rank pure compounds as well as influents and effluents of sewage treatment plants (STPs) according to estrogenic activity. In this adapted format, flow cytometric cell cycle analysis was used to allow evaluation of the MCF-7 cell proliferative effects after only 24 h of exposure. With an average EC{sub 50} value of 2 pM and CV of 22%, this assay appears as a sensitive and reproducible system for evaluation of estrogenic activity. Moreover, estrogenic responses of 17 pure compounds corresponded well, qualitatively and quantitatively, with other in vitro and in vivo estrogenicity screens, such as the classical E-screen (R{sup 2} = 0.98), the estrogen receptor (ER) binding (R{sup 2} = 0.84) and the ER transcription activation assay (R{sup 2} = 0.87). To evaluate the applicability of this assay for complex samples, influents and effluents of 10 STPs covering different treatment processes, were compared and ranked according to estrogenic removal efficiencies. Activated sludge treatment with phosphorus and nitrogen removal appeared most effective in eliminating estrogenic activity, followed by activated sludge, lagoon and filter bed. This is well in agreement with previous findings based on chemical analysis or biological activity screens. Moreover, ER blocking experiments indicated that cell proliferative responses were mainly ER mediated, illustrating that the complexity of the end point, cell proliferation, compared to other ER screens, does not hamper the interpretation of the results. Therefore, this study, among other E-screen studies
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International Nuclear Information System (INIS)
Vanparys, Caroline; Depiereux, Sophie; Nadzialek, Stephanie; Robbens, Johan; Blust, Ronny; Kestemont, Patrick; De Coen, Wim
2010-01-01
In vitro estrogenicity screens are believed to provide a first prioritization step in hazard characterization of endocrine disrupting chemicals. When applied to complex environmental matrices or mixture samples, they have been indicated valuable in estimating the overall estrogen-mimicking load. In this study, the performance of an adapted format of the classical E-screen or MCF-7 cell proliferation assay was profoundly evaluated to rank pure compounds as well as influents and effluents of sewage treatment plants (STPs) according to estrogenic activity. In this adapted format, flow cytometric cell cycle analysis was used to allow evaluation of the MCF-7 cell proliferative effects after only 24 h of exposure. With an average EC 50 value of 2 pM and CV of 22%, this assay appears as a sensitive and reproducible system for evaluation of estrogenic activity. Moreover, estrogenic responses of 17 pure compounds corresponded well, qualitatively and quantitatively, with other in vitro and in vivo estrogenicity screens, such as the classical E-screen (R 2 = 0.98), the estrogen receptor (ER) binding (R 2 = 0.84) and the ER transcription activation assay (R 2 = 0.87). To evaluate the applicability of this assay for complex samples, influents and effluents of 10 STPs covering different treatment processes, were compared and ranked according to estrogenic removal efficiencies. Activated sludge treatment with phosphorus and nitrogen removal appeared most effective in eliminating estrogenic activity, followed by activated sludge, lagoon and filter bed. This is well in agreement with previous findings based on chemical analysis or biological activity screens. Moreover, ER blocking experiments indicated that cell proliferative responses were mainly ER mediated, illustrating that the complexity of the end point, cell proliferation, compared to other ER screens, does not hamper the interpretation of the results. Therefore, this study, among other E-screen studies, supports the use of
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Elfer, Katherine; Sholl, Andrew; Miller, Christopher; Brown, J. Quincy
2015-07-01
Histological assessment of freshly removed tissue specimens requires accurate and fast analysis in clinical procedures such as diagnostic biopsy and surgical tumor resection. Current histological assessment methods are either time-consuming or damage the tissue beyond the ability to re-analyze post-procedure. We demonstrate a novel dual-stain fluorescent analogue to brightfield Hematoxylin and Eosin for in-procedure histopathology that is both time-efficient and preserves the analyzed tissue for later analysis. H&E-like images are created from the combination of DRAQ5 and Eosin applied to human prostate tissue and animal muscle tissue under confocal microscopy. D&E images are pseduocolored to match H&E coloring, showing near-identical features to brightfield H&E of the same tissue. The histological accuracy, short staining time, and tissue preservation aspects of this dual-stain technique demonstrates its potential to be adopted for use in point-of-care pathology.
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Cytometric Approach for Detection of Encephalitozoon intestinalis, an Emergent Agent▿
Barbosa, Joana; Rodrigues, Acácio Gonçalves; Pina-Vaz, Cidália
2009-01-01
Encephalitozoon intestinalis is responsible for intestinal disease in patients with AIDS and immunocompetent patients. The infectious form is a small spore that is resistant to water treatment procedures. Its detection is very important, but detection is very cumbersome and time-consuming. Our main objective was to develop and optimize a specific flow cytometric (FC) protocol for the detection of E. intestinalis in hospital tap water and human feces. To determine the optimal specific antibody (Microspor-FA) concentration, a known concentration of E. intestinalis spores (Waterborne, Inc.) was suspended in hospital tap water and stool specimens with different concentrations of Microspor-FA, and the tap water and stool specimens were incubated under different conditions. The sensitivity limit and specificity were also evaluated. To study spore infectivity, double staining with propidium iodide (PI) and Microspor-FA was undertaken. Distinct approaches for filtration and centrifugation of the stool specimens were used. E. intestinalis spores stained with 10 μg/ml of Microspor-FA at 25°C overnight provided the best results. The detection limit was 5 × 104 spores/ml, and good specificity was demonstrated. Simultaneous staining with Microspor-FA and PI ensured that the E. intestinalis spores were dead and therefore noninfectious. With the stool specimens, better spore recovery was observed with a saturated solution of NaCl and centrifugation at 1,500 × g for 15 min. A new approach for the detection of E. intestinalis from tap water or human feces that ensures that the spores are not viable is now available and represents an important step for the prevention of this threat to public health. PMID:19439525
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Jones, W. Jeremy; Grofcsik, Andras; Kubinyi, Miklos; Thomas, Daniel
2006-07-01
The pump and probe method concentration-modulated absorption spectroscopy has been applied for studying the triplet characteristics of dyes erythrosin B, rose bengal and eosin y in ethanol solutions. Using the 514.5 and 488 nm lines of an argon ion laser for pump and probe beams, respectively, photoinduced bleaching for erythrosin B and eosin y and photoinduced absorption for rose bengal have been observed. The theory developed for describing the interactions of dye molecules of strong triplet forming ability with continuous wave pump and probe beams of Gaussian beam profiles has been tested by analysing the probe power gain or loss at different positions along the optical path of the focused collinear beams. From the gain-modulation frequency curves triplet absorption cross sections and the lifetimes of the triplet states have been determined.
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International Nuclear Information System (INIS)
Yousuf, R.M.; Najam, I.A.
2009-01-01
In this work, three types of scintillation materials. Acridine orange, Eosin blue and Eosin yellow, were used to act as liquid scintillation detectors. They can be used to detect ionizing radiation, especially that of high Linear Energy Transfer (Let). This work determines the optimum concentration for each of the investigated materials to be 0.2 g/1 dissolved in methanol, added to a solution of Anthracene in Xylene of the concentration of 1.4 g/1 and a solution of POPOP in Xylene of the concentration of 0.2 g/1. All samples were irradiated using radioactive sources 241 Am, 137 Cs and 60 Co. (authors).
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Flow cytogenetic studies in chromosomes and whole cells for the detection of clastogenic effects
International Nuclear Information System (INIS)
Otto, F.J.; Oldiges, H.
1980-01-01
Flow cytometric measurements of the chromosomal DNA content have been used to develop a screening method for the detection of chemically- or physically-induced cytogenetic damage. The reproducibility of this flow cytogenetic assay was shown in a series of subcultures of a Chinese hamster cell clone. The accuracy and sensitivity was tested in cultures treated with chemical mutagens and x-rays. The clastogenic effectiveness was quantified and the dose-effect relationship was established by the increase of the coefficient of variation of the peak of the largest chromosome type in the flow histograms. Since structural chromosome aberrations cause an unequal division of the DNA at mitosis, it is expected that clastogenic effects can be detected also in whole cells of growing populations as an increased dispersion of the cellular DNA content. In order to test this feature, high resolution flow cytometric measurements were performed in x-irradiated hamster cells in vitro and mouse bone marrow cells in vivo
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Flow cytometric assessment of viability of lactic acid bacteria
Bunthof, C.J.; Bloemen, K.; Breeuwer, P.; Rombouts, F.M.; Abee, T.
2001-01-01
The viability of lactic acid bacteria is crucial for their applications as dairy starters and as probiotics. We investigated the usefulness of flow cytometry (FCM) for viability assessment of lactic acid bacteria. The esterase substrate carboxyfluorescein diacetate (cFDA) and the dye exclusion DNA
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Rodriguez, Eleazar; Azevedo, Raquel; Fernandes, Pedro; Santos, Conceição
2011-07-18
Chromium(VI) is recognized as the most toxic valency of Cr, but its genotoxicity and cytostaticity in plants is still poorly studied. In order to analyze Cr(VI) cyto- and gentotoxicity, Pisum sativum L. plants were grown in soil and watered with solutions with different concentrations of Cr up to 2000 mg/L. After 28 days of exposure, leaves showed no significant variations in either cell cycle dynamics or ploidy level. As for DNA damage, flow cytometric (FCM) histograms showed significant differences in full peak coefficient of variation (FPCV) values, suggesting clastogenicity. This is paralleled by the Comet assay results, showing an increase in DNA damage for 1000 and 2000 mg/L. In roots, exposure to 2000 mg/L resulted in cell cycle arrest at the G(2)/M checkpoint. It was also verified that under the same conditions 40% of the individuals analyzed suffered polyploidization having both 2C and 4C levels. DNA damage analysis by the Comet assay and FCM revealed dose-dependent increases in DNA damage and FPCV. Through this, we have unequivocally demonstrated for the first time in plants that Cr exposure can result in DNA damage, cell cycle arrest, and polyploidization. Moreover, we critically compare the validity of the Comet assay and FCM in evaluating cytogenetic toxicity tests in plants and demonstrate that the data provided by both techniques complement each other and present high correlation levels. In conclusion, the data presented provides new insight on Cr effects in plants in general and supports the use of the parameters tested in this study as reliable endpoints for this metal toxicity in plants. © 2011 American Chemical Society
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Flow lab.: flow visualization and simulation
International Nuclear Information System (INIS)
Park, Chung Kyun; Cho, Won Jin; Hahn, Pil Soo
2005-01-01
The experimental setups for flow visualization and processes identification in laboratory scale (so called Flow Lab.) has developed to get ideas and answer fundamental questions of flow and migration in geologic media. The setup was made of a granite block of 50x50cm scale and a transparent acrylate plate. The tracers used in this experiments were tritiated water, anions, and sorbing cations as well as an organic dye, eosine, to visualize migration paths. The migration plumes were taken with a digital camera as a function of time and stored as digital images. A migration model was also developed to describe and identify the transport processes. Computer simulation was carried out not only for the hydraulic behavior such as distributions of pressure and flow vectors in the fracture but also for the migration plume and the elution curves
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N-(1-Pyrenyl Maleimide Induces Bak Oligomerization and Mitochondrial Dysfunction in Jurkat Cells
Directory of Open Access Journals (Sweden)
Pei-Rong Huang
2015-01-01
Full Text Available N-(1-pyrenyl maleimide (NPM is a fluorescent reagent that is frequently used as a derivatization agent for the detection of thio-containing compounds. NPM has been shown to display a great differential cytotoxicity against hematopoietic cancer cells. In this study, the molecular mechanism by which NPM induces apoptosis was examined. Here, we show that treatment of Jurkat cells with NPM leads to Bak oligomerization, loss of mitochondrial membrane potential (Δψm, and release of cytochrome C from mitochondria to cytosol. Induction of Bak oligomerization appears to play a critical role in NPM-induced apoptosis, as downregulation of Bak by shRNA significantly prevented NPM-induced apoptosis. Inhibition of caspase 8 by Z-IETD-FMK and/or depletion of Bid did not affect NPM-induced oligomerization of Bak. Taken together, these results suggest that NPM-induced apoptosis is mediated through a pathway that is independent of caspase-8 activation.
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On the mechanism of photocatalytic reactions with eosin Y
Majek, Michal; Filace, Fabiana; von Wangelin, Axel Jacobi
2014-01-01
Summary A combined spectroscopic, synthetic, and apparative study has allowed a more detailed mechanistic rationalization of several recently reported eosin Y-catalyzed aromatic substitutions at arenediazonium salts. The operation of rapid acid–base equilibria, direct photolysis pathways, and radical chain reactions has been discussed on the basis of pH, solvent polarity, lamp type, absorption properties, and quantum yields. Determination of the latter proved to be an especially valuable tool...
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Kaastrup, Kaja; Aguirre-Soto, Alan; Wang, Chen; Bowman, Christopher N; Stansbury, Jeffery; Sikes, Hadley D
In conjunction with a tertiary amine coinitiator, eosin, a photoreducible dye, has been shown to successfully circumvent oxygen inhibition in radical photopolymerization reactions. However, the role of O 2 in the initiation and polymerization processes remains inconclusive. Here, we employ a UV-Vis/FT-NIR analytical tool for real-time, simultaneous monitoring of chromophore and monomer reactive group concentrations to investigate the eosin-activated photopolymerization of PEGDA-based hydrogels under ambient conditions. First, we address the challenges associated with spectroscopic monitoring of the polymerization of hydrogels using UV-Vis and FT-NIR, proposing metrics for quantifying the extent of signal loss from reflection and scattering, and showing their relation to microgelation and network formation. Second, having established a method for extracting kinetic information by eliminating the effects of changing refractive index and scattering, the coupled UV-Vis/FT-NIR system is applied to the study of eosin-activated photopolymerization of PEGDA in the presence of O 2 . Analysis of the inhibition time, rate of polymerization, and rate of eosin consumption under ambient and purged conditions indicates that regeneration of eosin in the presence of oxygen and consumption of oxygen occur via a nonchain process. This suggests that the uniquely high O 2 resilience is due to alternative processes such as energy transfer from photo-activated eosin to oxygen. Uncovering the intricacies of the role of O 2 in eosin-mediated initiation aids the design of O 2 resistant free radical polymerization systems relevant to photonics, optoelectronics, biomaterials, and biosensing.
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Energy Technology Data Exchange (ETDEWEB)
Li, Yuexiang; Guo, Miaomiao; Peng, Shaoqin [Department of Chemistry, Nanchang University, Nanchang 330031 (China); Lu, Gongxuan; Li, Shuben [State Key Laboratory for Oxo Synthesis and Selective Oxidation, Lanzhou Institute of Chemical Physics, Chinese Academy of Sciences, Lanzhou 730000 (China)
2009-07-15
An efficient visible-light active photocatalyst of multilayer-Eosin Y-sensitized TiO{sub 2} is prepared through linkage of Fe{sup 3+} between not only TiO{sub 2} and Eosin Y but also different Eosin Y molecules to form three-dimensional polymeric dye structure. The multilayer-dye-sensitized photocatalyst is found to have high light harvesting efficiency and photocatalytic activity for hydrogen evolution under visible light irradiation ({lambda} > 420 nm). On the optimum conditions (1:1 initial molar ratio of Eosin Y to Fe(NO{sub 3}){sub 3}, initial 10 x 10{sup -3} M Eosin Y, and 1.0 wt% Pt deposited by in situ photoreduction), its maximal apparent quantum yield for hydrogen evolution is 19.1% from aqueous triethanolamine solution (TEOA aq). The present study highlights linking between dye molecules via metal ions as a general way to develop efficient visible-light photocatalyst. (author)
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Energy Technology Data Exchange (ETDEWEB)
Vignesh, K. [P.G. and Research Department of Chemistry, Thiagarajar College, Madurai, Tamilnadu 625009 (India); Suganthi, A., E-mail: suganthiphd09@gmail.com [P.G. and Research Department of Chemistry, Thiagarajar College, Madurai, Tamilnadu 625009 (India); Rajarajan, M., E-mail: rajarajan_1962@yahoo.com [Department of Chemistry, C.P.A. College, Bodinayakanur, Tamilnadu 626513 (India); Sakthivadivel, R. [P.G. and Research Department of Chemistry, Thiagarajar College, Madurai, Tamilnadu 625009 (India)
2012-03-01
Hesperidin a flavanoid, modified TiO{sub 2} nanoparticles (Hes-TiO{sub 2}) was synthesized to improve the visible light driven photocatalytic performance of TiO{sub 2}. The synthesized nanoparticles were characterized by UV-visible diffuse reflectance spectroscopy (UV-vis-DRS), FT-IR, powder X-ray diffraction (XRD) and scanning electron microscopy (SEM). The photocatalytic activity of Hes-TiO{sub 2} was investigated based on the decolorization of eosin-Y under visible light irradiation. Hes-TiO{sub 2} showed high efficiency for the decolorization of eosin-Y. The influences of various reaction parameters like effect of pH, catalyst dosage and initial dye concentration on the photocatalytic efficiency were investigated. The adsorption of eosin-Y on Hes-TiO{sub 2} was found favorable by the Langmuir approach. The removal percentage of chemical oxygen demand (COD) was determined to evaluate the mineralization of eosin-Y during photodecolorization. Based on the intermediates obtained in the GC-MS spectroscopic technique, a probable degradation mechanism has been proposed.
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Flow cytometric chromosome sorting in plants: The next generation
Czech Academy of Sciences Publication Activity Database
Vrána, Jan; Šimková, Hana; Kubaláková, Marie; Čihalíková, Jarmila; Doležel, Jaroslav
2012-01-01
Roč. 57, č. 3 (2012), s. 331-337 ISSN 1046-2023 R&D Projects: GA ČR GAP501/10/1740 Grant - others:GA MŠk(CZ) ED0007/01/01 Program:ED Institutional research plan: CEZ:AV0Z50380511 Keywords : Chromosome sorting * Flow cytometry * Fluorescence in situ hybridization Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 3.641, year: 2012
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Directory of Open Access Journals (Sweden)
V.O. Njoku
2013-05-01
Full Text Available This paper reports the use of powdered activated carbon (PAC and raw coal fly ash (RFA in the removal of eosin dye from aqueous solution in batch processes. Operational parameters such as contact time, initial dye concentration, pH and temperature were investigated. Adsorption equilibrium was established in 120 min for the two adsorbents. Langmuir and Freundlich isotherms were used to fit the adsorption data. Langmuir model gave the best fit in both cases. The adsorption capacities of PAC and RFA were found to be 62.28 mg/g and 43.48 mg/g, respectively. The highest percentage of eosin dye removal for both PAC (98% and RFA (90% was observed at pH 2. Pseudo first-order and pseudo second-order kinetic models were used to fit the adsorption data. Pseudo second-order kinetic model gave the best description of the adsorption of eosin dye onto the two adsorbents. Thermodynamic parameters, ΔH0, ΔS0 and ΔG0 confirmed the physical nature, spontaneity and the endothermic nature of the adsorption process. A regeneration technique and a process calculation for evaluating the adsorbent dose required were carried out. This study has shown that RFA is a good alternative adsorbent in the removal of eosin dye from aqueous solution.
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Ghosh, Sujit Kumar; Pal, Anjali; Nath, Sudip; Kundu, Subrata; Panigrahi, Sudipa; Pal, Tarasankar
2005-08-01
Gold nanoparticles of variable sizes have been exploited to study their influence on the absorption and emission spectral characteristics of eosin, a fluorescent dye. It has been found that smaller particles of gold stimulate J-aggregation of eosin on the surface of metal particles whereas larger particles cannot induce any kind of aggregation amongst the dye molecules. The size regime dependence of the gold nanoparticles has been attributed to the intercluster interactions induced by the dye molecules for smaller gold nanoparticles and consequently, close packing of the dye molecules around the gold surface engenders intermolecular interactions amongst the dye molecules leading to dimerization.
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Cerezo, Maria I.; Linden, Matthew; Agusti, Susana
2017-01-01
Polycyclic aromatic hydrocarbons are very important components of oil pollution. These pollutants tend to sorb to cell surfaces, exerting toxic effects on organisms. Our study developed a flow cytometric method for the detection of PAHs sorbed
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Flow cytometric applications of tumor biology: prospects and pitfalls
International Nuclear Information System (INIS)
Raju, M.R.; Johnson, T.S.; Tokita, N.; Gillette, E.L.
1979-01-01
A brief review of cytometry instrumentation and its potential applications in tumor biology is presented using our recent data. Age-distribution measurements of cells from spontaneous dog tumors and cultured cells after exposure to x rays, alpha particles, or adriamycin are shown. The data show that DNA fluorescence measurements have application in the study of cell kinetics after either radiation or drug treatment. Extensive and careful experimentation is needed to utilize the sophisticated developments in flow cytometry instrumentation
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Flow cytometric applications to tumour biology: prospects and pitfalls
International Nuclear Information System (INIS)
Raju, M.R.; Johnson, T.S.; Tokita, N.; Gillette, E.L.
1980-01-01
A brief review of cytometry instrumentation and its potential applications in tumour biology is presented. DNA distribution measurements of cells from spontaneous dog tumours and cultured cells after exposure to X-rays, alpha particles or adriamycin are shown. The data show that DNA fluorescence measurements have application in the study of cell kinetics after either radiation or drug treatment. Extensive and careful experimentation is needed, however, to utilize the sophisticated developments in flow cytometry instrumentation. (author)
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Automated flow cytometric analysis across large numbers of samples and cell types.
Chen, Xiaoyi; Hasan, Milena; Libri, Valentina; Urrutia, Alejandra; Beitz, Benoît; Rouilly, Vincent; Duffy, Darragh; Patin, Étienne; Chalmond, Bernard; Rogge, Lars; Quintana-Murci, Lluis; Albert, Matthew L; Schwikowski, Benno
2015-04-01
Multi-parametric flow cytometry is a key technology for characterization of immune cell phenotypes. However, robust high-dimensional post-analytic strategies for automated data analysis in large numbers of donors are still lacking. Here, we report a computational pipeline, called FlowGM, which minimizes operator input, is insensitive to compensation settings, and can be adapted to different analytic panels. A Gaussian Mixture Model (GMM)-based approach was utilized for initial clustering, with the number of clusters determined using Bayesian Information Criterion. Meta-clustering in a reference donor permitted automated identification of 24 cell types across four panels. Cluster labels were integrated into FCS files, thus permitting comparisons to manual gating. Cell numbers and coefficient of variation (CV) were similar between FlowGM and conventional gating for lymphocyte populations, but notably FlowGM provided improved discrimination of "hard-to-gate" monocyte and dendritic cell (DC) subsets. FlowGM thus provides rapid high-dimensional analysis of cell phenotypes and is amenable to cohort studies. Copyright © 2015. Published by Elsevier Inc.
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Ginting, R. T.; Yap, C. C.; Yahaya, M.; Fauzia, V.; Salleh, M. M.
2013-04-01
The active layer spin coating speed dependence of the performance of inverted organic solar cells (OSCs) based on Eosin-Y-coated ZnOnanorods has been investigated. An active layer consisted of poly(2-methoxy-5-(2'-ethyl)-hexyloxy-p-phenylenevinylene) (MEH-PPV) as donor and phenyl-c61-butyric acid methyl ester (PCBM) as acceptor was employed, whereas ZnO nanorods were utilized as electron transporting layer. The active layer was deposited on top of Eosin-Y-coated ZnO nanorods with various spin coating speeds (1000-4000 rpm). Inverted OSCs with a structure of FTO/Eosin-Y-coated ZnO nanorods/MEH-PPV:PCBM /Ag were characterized through the current density-voltage (J-V) measurement under illumination intensity of 100 mW/cm2. Based on the investigation, the short circuit current density (Jsc) and the power conversion efficiency (PCE) enhanced significantly, where as fill factor slightly increased with spin coating speed. The two-diode equivalent model was found to fit the experimental J-V curves very well. The optimum PCE of 1.18 ± 0.07% was achieved at the highest spin coating speed of 4000 rpm, as a result of the decrement of diffusion current density (Jdiff), recombination current density (Jrec), and ideality factor, thus further confirms the strong built-in electric field in thinner photoactive layer.
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Lv, Zhengxian; Sun, Zhiwei; Song, Cuihua; Lu, Shuaimin; Chen, Guang; You, Jinmao
2016-12-01
A sensitive and background-free pre-column derivatization method for the determination of thiol compounds using metal-organic framework material (MOF-5) as dispersive solid-phase extraction (DSPE) adsorbent followed by high-performance liquid chromatography fluorescence detection (HPLC-FLD) has been developed. In this paper, a novel labeling reagent, carbazole-9-ethyl-2-maleimide(CAEM), was synthesized and reacted with thiols at 40°C for 10min in the presence of PBS buffer (0.02mol/L, pH 7.5). Interestingly, CAEM itself had no fluorescence, while its derivatives exhibited intense fluorescence with an excitation maximum at λ ex 274nm and an emission maximum at λ em 363nm, which greatly reduced the background interference and improved the sensitivity of the method. Furthermore, the MOF-5 was prepared and used as DSPE adsorbent for the selective adsorption of thiols from wastewater sample. Under the optimized experimental conditions, an excellent linearity for all analytes over their concentration ranges of 0.01-1.0μmol/L (R 2 >0.9986)were obtained with the limit of detection (LOD) ranging from 8 to 17.1pmol/L for nine tested thiols. The feasibility of this method for the determination of thiols in wastewater samples had been evaluated and satisfactory average recoveries (n=3) were achieved with the range of 86.6-98.5%. Copyright © 2016 Elsevier B.V. All rights reserved.
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Derayea, Sayed M.; Omar, Mahmoud A.; Abu-hassan, Ahmed A.
2018-03-01
A simple, sensitive and non-extractive spectrofluorimetric method has been developed and validated for the determination of two psychoanaleptic drugs, atomoxetine and fluvoxamine, in pure forms and pharmaceutical dosage forms. The proposed method is based on the formation of binary complexes between eosin Y and the studied drugs in the presence of a Teorell-Stenhagen buffer. The quenching of the native fluorescence of eosin Y due to complex formation with the studied drugs was measured spectrofluorimetrically at 545 nm after excitation at 302 nm. At the optimum reaction conditions, the fluorescence quenching values (ΔF) and concentrations were rectilinear over the concentration ranges of 0.2-2.2 and 0.3-2.2 µg ml-1 for atomoxetine and fluvoxamine, respectively. The developed method was successfully applied for the determination of the studied drugs in their pharmaceutical formulations with average percentage recoveries of 100.13 ± 0.66 and 99.69 ± 0.44 for atomoxetine and fluvoxamine, respectively (n = 5), without interference from common excipients.
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On the mechanism of photocatalytic reactions with eosin Y
Directory of Open Access Journals (Sweden)
Michal Majek
2014-04-01
Full Text Available A combined spectroscopic, synthetic, and apparative study has allowed a more detailed mechanistic rationalization of several recently reported eosin Y-catalyzed aromatic substitutions at arenediazonium salts. The operation of rapid acid–base equilibria, direct photolysis pathways, and radical chain reactions has been discussed on the basis of pH, solvent polarity, lamp type, absorption properties, and quantum yields. Determination of the latter proved to be an especially valuable tool for the distinction between radical chain and photocatalytic reactions.
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Ghosh, Soumen; Roscioli, Jerome D.; Beck, Warren F.
2014-06-01
We have employed 2D electronic photon echo spectroscopy to study intramolecular charge-transfer dynamics in eosin B. After preparation of the first excited singlet state (S_1) with 40-fs excitation pulses at 520 nm, the nitro group (--NO_2) in eosin B undergoes excited state torsional motion towards a twisted intramolecular charge transfer (TICT) state. As the viscosity of the surrounding solvent increases, the charge-transfer rate decreases because the twisting of the --NO_2 group is hindered. These conclusions are supported by the time evolution of the 2D spectrum, which provides a direct measure of the the ground-to-excited-state energy gap time-correlation function, M(t). In comparison to the inertial and diffusive solvation time scales exhibited by eosin Y, which lacks the nitro group, the M(t) function for eosin B exhibits under the same conditions an additional component on the 150-fs timescale that arises from quenching of the S_1 state by crossing to the TICT state. These results indicate that 2D electronic spectroscopy can be used as a sensitive probe of the rate of charge transfer in a molecular system and of the coupling to the motions of the surrounding solvent. (Supported by grant DE-SC0010847 from the Department of Energy, Office of Basic Energy Sciences, Photosynthetic Systems program.)
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International Nuclear Information System (INIS)
Machherndl-Spandl, S; Suessner, S; Danzer, M; Proell, J; Gabriel, C; Lauf, J; Sylie, R; Klein, H-U; Béné, M C; Weltermann, A; Bettelheim, P
2013-01-01
Special attention has recently been drawn to the molecular network of different genes that are responsible for the development of erythroid cells. The aim of the present study was to establish in detail the immunophenotype of early erythroid cells and to compare the gene expression profile of freshly isolated early erythroid precursors with that of the CD34-positive (CD34 + ) compartment. Multiparameter flow cytometric analyses of human bone marrow mononuclear cell fractions (n=20) defined three distinct early erythroid stages. The gene expression profile of sorted early erythroid cells was analyzed by Affymetrix array technology. For 4524 genes, a differential regulation was found in CD105-positive erythroid cells as compared with the CD34 + progenitor compartment (2362 upregulated genes). A highly significant difference was observed in the expression level of genes involved in transcription, heme synthesis, iron and mitochondrial metabolism and transforming growth factor-β signaling. A comparison with recently published data showed over 1000 genes that as yet have not been reported to be upregulated in the early erythroid lineage. The gene expression level within distinct pathways could be illustrated directly by applying the Ingenuity software program. The results of gene expression analyses can be seen at the Gene Expression Omnibus repository
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Flow cytometric DNA ploidy analysis of ovarian granulosa cell tumors
D. Chadha; C.J. Cornelisse; A. Schabert (A.)
1990-01-01
textabstractAbstract The nuclear DNA content of 50 ovarian tumors initially diagnosed as granulosa cell tumors was measured by flow cytometry using paraffin-embedded archival material. The follow-up period of the patients ranged from 4 months to 19 years. Thirty-eight tumors were diploid or
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Cell cycle evaluation of granulosa cells in the {gamma}-irradiated mouse ovarian follicles
Energy Technology Data Exchange (ETDEWEB)
KIm, Jin Kyu; Lee, Chang Joo; Lee, Young Keun [Korea Atomic Energy Research Institute, Taejon (Korea, Republic of); Song, Kang Won; Yoon, Yong Dal [Hanyang Univ., Seoul (Korea, Republic of)
1999-03-01
This study was carried out to evaluate the biochemical and morphological effects of ionizing radiation on mouse ovarian follicles. Immature mice (ICR, 3 week-old) were irradiated with a dose of LD{sub 80(30)} at KAERI. The ovaries were collected after 6 hours, 12 hours, 1 day, and 2 days post irradiation. With the morphological basis of the histological staining with hematoxylin-eosin, immunohistochemical preparation using in situ 3'-end labeling was evaluated. Flow cytometric evaluation of DNA extracted from the whole ovary was performed. The percentage of A{sub 0} (subpopulation of cells with degraded DNA and with lower DNA fluorescence than G{sub 0}/G{sub 1} cells), apoptotic, cells in the cell cycle was significantly higher in the irradiated group than in the control group. The number of in situ 3'-end labeled follicles increased at 6 hours post irradiation. All the analyses represented that the ionizing radiation-induced follicular atresia was taken place via an apoptotic degeneration. Such a degeneration underwent very fast and acutely. Therefore, it is concluded that the radiation-induced follicular degeneration is, like the spontaneous atresia, mediated by an acute apoptosis of follicular granulosa cells. Flow cytometric evaluation of cell cycles can make the role for quantifying the atretic follicles and understanding the mechanism of the radiation-induced cell death.
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Zhou, Fangbin; Zhou, Yaying; Yang, Ming; Wen, Jinli; Dong, Jun; Tan, Wenyong
2018-01-01
Circulating endothelial cells (CECs) and their subpopulations could be potential novel biomarkers for various malignancies. However, reliable enumerable methods are warranted to further improve their clinical utility. This study aimed to optimize a flow cytometric method (FCM) assay for CECs and subpopulations in peripheral blood for patients with solid cancers. An FCM assay was used to detect and identify CECs. A panel of 60 blood samples, including 44 metastatic cancer patients and 16 healthy controls, were used in this study. Some key issues of CEC enumeration, including sample material and anticoagulant selection, optimal titration of antibodies, lysis/wash procedures of blood sample preparation, conditions of sample storage, sufficient cell events to enhance the signal, fluorescence-minus-one controls instead of isotype controls to reduce background noise, optimal selection of cell surface markers, and evaluating the reproducibility of our method, were integrated and investigated. Wilcoxon and Mann-Whitney U tests were used to determine statistically significant differences. In this validation study, we refined a five-color FCM method to detect CECs and their subpopulations in peripheral blood of patients with solid tumors. Several key technical issues regarding preanalytical elements, FCM data acquisition, and analysis were addressed. Furthermore, we clinically validated the utility of our method. The baseline levels of mature CECs, endothelial progenitor cells, and activated CECs were higher in cancer patients than healthy subjects ( P technical issues found in previously published assays and validated the reproducibility and sensitivity of our proposed method. Future work is required to explore the potential of our optimized method in clinical oncologic applications.
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International Nuclear Information System (INIS)
Coureuil, M.; Tavernier, M.; Barroca, V.; Fouchet, P.; Allemand, I.; Ugolin, N.; Chevillard, S.
2010-01-01
Spermatogonia- stem cells and progenitors of adult spermatogenesis- are killed through a p53-regulated apoptotic process after γ-irradiation but the death effectors are still poorly characterized. Our data demonstrate that both intrinsic and extrinsic apoptotic pathways are involved, and especially that spermatogonia can be split into two main populations, according to apoptotic effectors. Following irradiation both Dr5 and Puma genes are up-regulated in the α 6 -integrin-positive Side Population (SP) fraction, which is highly enriched in spermatogonia. Flow cytometric analysis confirms an increased number of Dr5-expressing SP cells, and Puma-β isoform accumulates in α 6 -integrin positive cellular extracts, enriched in spermatogonia. Trail -/- or Puma -/- spermatogonia display a reduced sensitivity to radiation-induced apoptosis. The TUNEL kinetics strongly suggest that the extrinsic and intrinsic pathways, via Trail/Dr5 and Puma respectively, could be engaged in distinct subpopulations of spermatogonia. Indeed flow cytometric studies show that Dr5 receptor is constitutively present on more than half of the undifferentiated progenitors (Kit - α 6 + SP) and half of the differentiated ones (Kit + α 6 + SP). In addition after irradiation, Puma is not detected in the Dr5-positive cellular fraction isolated by immuno-magnetic purification, while Puma is present in the Dr5-negative cell extracts. In conclusion, adult testicular progenitors are divided into distinct sub-populations by apoptotic effectors, independently of progenitor types (immature Kit-negative versus mature Kit-positive), underscoring differential radiosensitivities characterizing the stem cell/progenitors compartment. (authors)
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Directory of Open Access Journals (Sweden)
C. Gaina
2012-02-01
Full Text Available New thermo-reversible networks were obtained from poly(vinyl furfural and multifunctional maleimide monomers by Diels-Alder (DA and retro-DA reactions. The poly(vinyl furfural having acetalization degree of 15 and 25% were obtained by the acid-catalyzed homogenous acetalization of poly(vinyl alcohol with 2-furfural in a nonaqueous media. The thermal and viscoelastic behaviour of the cross-linked materials have been studied via differential scanning calorimetry, dynamic mechanical analysis and thermogravimetric analysis. The networks exhibit considerable swelling in those organic solvents that dissolve both poly(vinyl furfural and bismaleimides; by heating in aprotic dipolar solvents at 150°C, they become soluble.
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Photodegradation of Eosin Y Using Silver-Doped Magnetic Nanoparticles.
Alzahrani, Eman
2015-01-01
The purification of industrial wastewater from dyes is becoming increasingly important since they are toxic or carcinogenic to human beings. Nanomaterials have been receiving significant attention due to their unique physical and chemical properties compared with their larger-size counterparts. The aim of the present investigation was to fabricate magnetic nanoparticles (MNPs) using a coprecipitation method, followed by coating with silver (Ag) in order to enhance the photocatalytic activity of the MNPs by loading metal onto them. The fabricated magnetic nanoparticles coated with Ag were characterised using different instruments such as a scanning electron microscope (SEM), transmission electron microscopy (TEM), energy-dispersive X-ray (EDAX) spectroscopy, and X-ray diffraction (XRD) analysis. The average size of the magnetic nanoparticles had a mean diameter of about 48 nm, and the average particle size changed to 55 nm after doping. The fabricated Ag-doped magnetic nanoparticles were used for the degradation of eosin Y under UV-lamp irradiation. The experimental results revealed that the use of fabricated magnetic nanoparticles coated with Ag can be considered as reliable methods for the removal of eosin Y since the slope of evaluation of pseudo-first-order rate constant from the slope of the plot between ln(C o /C) and the irradiation time was found to be linear. Ag-Fe3O4 nanoparticles would be considered an efficient photocatalyst to degrade textile dyes avoiding the tedious filtration step.
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Photodegradation of Eosin Y Using Silver-Doped Magnetic Nanoparticles
Directory of Open Access Journals (Sweden)
Eman Alzahrani
2015-01-01
Full Text Available The purification of industrial wastewater from dyes is becoming increasingly important since they are toxic or carcinogenic to human beings. Nanomaterials have been receiving significant attention due to their unique physical and chemical properties compared with their larger-size counterparts. The aim of the present investigation was to fabricate magnetic nanoparticles (MNPs using a coprecipitation method, followed by coating with silver (Ag in order to enhance the photocatalytic activity of the MNPs by loading metal onto them. The fabricated magnetic nanoparticles coated with Ag were characterised using different instruments such as a scanning electron microscope (SEM, transmission electron microscopy (TEM, energy-dispersive X-ray (EDAX spectroscopy, and X-ray diffraction (XRD analysis. The average size of the magnetic nanoparticles had a mean diameter of about 48 nm, and the average particle size changed to 55 nm after doping. The fabricated Ag-doped magnetic nanoparticles were used for the degradation of eosin Y under UV-lamp irradiation. The experimental results revealed that the use of fabricated magnetic nanoparticles coated with Ag can be considered as reliable methods for the removal of eosin Y since the slope of evaluation of pseudo-first-order rate constant from the slope of the plot between ln(Co/C and the irradiation time was found to be linear. Ag-Fe3O4 nanoparticles would be considered an efficient photocatalyst to degrade textile dyes avoiding the tedious filtration step.
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Directory of Open Access Journals (Sweden)
Soad S. Abd El-Hay
2016-09-01
Full Text Available A simple and sensitive spectrophotometric method has been established for the determination of clemastine hydrogen fumarate (I, desloratadine (II, losartan potassium(III and moxepril HCl(IV based on binary complex formation with eosin. The method does not involve solvent extraction through the use of a non-ionic surfactant (methylcellulose. The color of the produced complex was measured at 552, 549 nm for (I, (II while was measured at 540 nm for (III and (IV. Appropriate conditions were established for the color reaction between eosin and the studied drugs to obtain maximum sensitivity. Under the proposed conditions, the method is applicable over concentration range of 1.25–11.25, 0.31–2.81, 2.5–20 and 1.25–15 μg/ml for (I, (II, (III and (IV, respectively. The molar absorptivity (ε, sandell sensitivity, detection (LOD and quantitation limits (LOQ are calculated. Unlike other reported ion-pair techniques, the suggested methods have the advantage of being applicable for the determination of the four drugs in their pharmaceutical dosage forms without prior extraction with excellent recoveries.
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Energy Technology Data Exchange (ETDEWEB)
Jones, G. II; Feng, Z.; Oh, C. [Boston Univ., MA (United States)
1995-03-23
The Xanthene dye eosin Y has been modified via a thiohydantoin link to the amine terminus of the amino acid L-tyrosine. Photochemical electron transfer involving the singlet state of the dye and the attached phenol-containing residue led to a reduction in eosin fluorescence quantum yield and lifetime for aqueous solutions at elevated pH. The conjugate provided an electron transfer product of relatively long lifetime (1 {mu}s range) observed by flash photolysis of solutions at pH 12.0, conditions under which the tyrosine moiety is ionized. The effects of binding of the conjugate in the polymer poly(vinylpyrrolidone) (PVP) on the rates of electron transfer of species of different charge type were examined. 30 refs., 5 figs., 1 tab.
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Directory of Open Access Journals (Sweden)
B. L. Hiran
2007-01-01
Full Text Available This article describes the synthesis and characterization of homopolymer (H-BCPM of N-[4-N'-(benzylamino-carbonyl phenyl] maleimide (N-BACPMI and copolymer (C-BCPM of N-BACPMI with n-butyl acrylate (BA. The new monomer was synthesized from p-aminobenzoic acid, maleic anhydride and benzylamine. The homopolymerization of N-BACPMI is initiated by free radical using AIBN in THF solvent at 65°C. Radical copolymerization of N-BACPMI with BA, initiated by AIBN, was performed in THF solvent using equimolar amount. Effect of the different free radical initiator AIBN, BPO and solvents p-Dioxane, THF, DMF and DMSO was studied. Homopolymer and Copolymer were characterized by intrinsic viscosity, solubility test, FT-IR, 1H-NMR spectral analysis and elemental analysis. Thermal behaviour was studied by Thermo gravimetric analysis.
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Hirschsprung's disease diagnosis: Comparison of immunohistochemical, hematoxilin and eosin staining
Memarzadeh, Mehrdad; Talebi, Ardeshir; Edalaty, Masod; Hosseinpour, Mehrdad; Vahidi, Nasrin
2009-01-01
Background: The diagnosis of Hirschsprung's disease (HD) is based on the absence of ganglion cells. In hemotoxilin and eosin (H and E) as well as acetylcholine esterase staining there are limitations in the diagnosis of immature ganglion cells in neonates. Methods: In this prospective study, 54 biopsies taken from suspected HD patients (five mucosal specimens and 49 full thickness specimens) were studied. In the laboratory, after preparing sections of paraffin embedded tissues, H and E staini...
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Sreethawong, Thammanoon; Junbua, Chompoonuch; Chavadej, Sumaeth
Sensitized photocatalytic production of hydrogen from water splitting is investigated under visible light irradiation over mesoporous-assembled titanium dioxide (TiO 2) nanocrystal photocatalysts, without and with Pt loading. The photocatalysts are synthesized by a sol-gel process with the aid of a structure-directing surfactant and are characterized by N 2 adsorption-desorption analysis, X-ray diffraction, UV-vis spectroscopy, scanning electron microscopy, transmission electron microscopy and energy-dispersive X-ray analysis. The dependence of hydrogen production on the type of TiO 2 photocatalyst (synthesized mesoporous-assembled and commercial non-mesoporous-assembled TiO 2 without and with Pt loading), the calcination temperature of the synthesized photocatalyst, the sensitizer (Eosin Y) concentration, the electron donor (diethanolamine) concentration, the photocatalyst dosage and the initial solution pH is systematically studied. The results show that in the presence of the Eosin Y sensitizer, the Pt-loaded mesoporous-assembled TiO 2 synthesized by a single-step sol-gel process and calcined at 500 °C exhibits the highest photocatalytic activity for hydrogen production from a 30 vol.% diethanolamine aqueous solution with dissolved 2 mM Eosin Y. Moreover, the optimum photocatalyst dosage and initial solution pH for the maximum photocatalytic activity for hydrogen production are 3.33 g dm -3 and 11.5, respectively.
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Eosin Y as a Direct Hydrogen Atom Transfer Photocatalyst for the Functionalization of C-H Bonds.
Fan, Xuan-Zi; Rong, Jia-Wei; Wu, Hao-Lin; Zhou, Quan; Deng, Hong-Ping; Tan, Jin Da; Xue, Cheng-Wen; Wu, Li-Zhu; Tao, Hai-Rong; Wu, Jie
2018-05-02
Eosin Y, a well-known economical alternative to metal catalysts in visible-light-driven single-electron transfer-based organic transformations, can behave as an effective direct hydrogen atom transfer catalyst for C-H activation. Using the alkylation of C-H bonds with electron-deficient alkenes as a model study revealed an extremely broad substrate scope, enabling easy access to a variety of important synthons. This eosin Y-based photocatalytic hydrogen atom transfer strategy is promising for diverse functionalization of a wide range of native C-H bonds in a green and sustainable manner. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
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Switchgrass (Panicum virgatum L.), a native perennial dominant of the prairies of North America, has been targeted as a model herbaceous species for biofeedstock development. A flow-cytometric survey of a core set of 11 primarily upland polyploid switchgrass accessions indicated that there was con...
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International Nuclear Information System (INIS)
Spano, M.; Leonardi, M.; Cordelli, E.
1991-01-01
Since DNA is the main cellular target of ionizing irradiation, methods fit for analyzing DNA alterations should be able to discern irradiated versus control cells. Flow cytometry allows the rapid measurement of DNA content of single chromosomes or cell nuclei at very high resolution on a statistically significant sample. Alterations of chromatine structure can also be analyzed by flow cytometry. Briefly, evaluation of in situ DNA resistance to denaturation can be evaluated by flow cytometric analysis of different staining pattern of single versus double strange regions of DNA. In the present work both approaches were used with the aim to recognize cells derived from an irradiated sample of breast chicken. Although flow cytometry has been demonstrated to be a useful tool to detect DNA alterations and has been widely used to detect damages on DNA induced by several physical and chemical agents, it was unable to detect clastogenic effects induced by electrons on DNA of chicken breast cells. Heavily irradiated nuclei, even if challenged by denaturating treatments that partially collapse chromatine organization, do not present differences from non irradiated samples after flow cytometric DNA content measurement. (16 refs)
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Mayani, Vishal J; Mayani, Suranjana V; Kim, Sang Wook
2017-08-03
Up to now, a very few catalysts have been developed approaching the heterogeneous catalytic degradation of Eosin Y and Chromotrope 2R dyes (Acid Red 29). The present study provides a complete perspective of recyclable nanocomposite Au(Salen)@CC for catalytic degradation of hazardous water pollutant dyes viz., Eosin Y & Chromotrope 2R using mild reaction conditions. New gold Salen complex doped carbon nanocomposite Au(Salen)@CC was developed by easy methodology using nano carbon cage (CC) prepared from low-priced Pyrolysis fuel oil (PFO) residue based Pitch. The UV-Vis adsorption spectroscopy results of Eosin Y and Chromotrope 2R dyes indicated complete degradation into acidic compounds which can be further mineralized to CO 2 and H 2 O under mild reaction conditions. The heterogeneous catalyst recycled and reused successfully for four repeated experiments without loss in its adequate performance. This new sustainable and eco-friendly catalyst delivered significant degradation activity compared to existing reports and it can be further utilized for new multifunctional applications such as, radiopharmaceutical activities, heterogeneous catalysis and chiral resolution.
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Hao, Xia; Lei, Jing Lei; Li, Nian Bing; Luo, Hong Qun
2014-12-10
A simple and effective method for the detection of electrochemically inactive sodium dodecyl sulfate (SDS) has been designed, based on different binding affinity of polyethyleneimine (PEI) toward electrochemically active eosin Y and electrochemically inactive SDS. The stronger binding affinity of the PEI toward SDS than eosin Y results in the decrease of the redox peak current of surface confined eosin Y and provides a quantitative readout for the SDS. The difference in value of the cathodic peak current showed a linear relationship with SDS concentration in a concentration range from 1 to 40 μg mL(-1), and a detection limit of 0.9 μg mL(-1) for SDS was obtained. Furthermore, the method has been successfully applied to the detection of SDS in real samples. The developed approach provided a simple and reliable detection for SDS and might have potential applications in electrochemical methods for inactive molecules. Copyright © 2014 Elsevier B.V. All rights reserved.
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Eosin-Y and N3-Dye sensitized solar cells (DSSCs) based on novel nanocoral TiO2: A comparative study
International Nuclear Information System (INIS)
Mali, Sawanta S.; Betty, C.A.; Bhosale, P.N.; Patil, P.S.
2012-01-01
Highlights: ► Synthesis of novel TiO 2 nanocorals by hydrothermal route. ► Comparative study of Eosin-Y and N3-Dyes. ► The efficiency of N3-Dye is 66% improved to that of Eosin-Y. ► The highest solar energy conversion efficiency 2.37% was observed for N3-Dye loaded DSSCs. - Abstract: Titanium oxide (TiO 2 ) nanocorals containing nanopolyps have been synthesized by a cost effective hydrothermal route directly on fluorine doped tin oxide (FTO) coated conducting glass substrates. The morphological features and physical properties of TiO 2 films were investigated by field-emission scanning electron microscopy, high resolution transmission electron microscopy, X-ray diffraction, Fourier transform Raman spectroscopy, room temperature photoluminescence spectroscopy and X-ray photoelectron spectroscopy. The surface morphology revealed the formation of TiO 2 corals having nanosized (30–40 nm) polyps. Further, we have studied its dye sensitized solar cell performance by using Eosin-Y and N3-Dye. The results indicate that the photoconversion efficiency of N3-Dye is 66% compared to that of Eosin-Y. The highest solar energy conversion efficiency of 2.37% was observed for N3-Dye loaded DSSCs.
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Bryukhanov, V. V.; Minaev, B. M.; Tsibul'nikova, A. V.; Slezhkin, V. A.
2015-07-01
We have studied exchange processes in contact complexes of triplet eosin molecules with oxygen molecules in the triplet (3Σ{/g -}) and singlet (1Δ g ) states in thin polyvinylbutyral films in the presence of gold nanoparticles. Upon resonant excitation of surface plasmons in gold nanoparticles into the absorption band of eosin molecules-singlet oxygen sensitizers-we have obtained an increase in the intensity of the delayed fluorescence and an increase in the lifetime of the dye with simultaneous quenching of the luminescence of singlet oxygen. The kinetics of the delayed fluorescence of the dye as a result of singlet-triplet annihilation of triplet eosin molecules with singlet oxygen molecules has been investigated. To compare theoretical and experimental data, we have numerically simulated energy transfer processes. Rate constants of energy transfer and of singlet-triplet annihilation, as well as quenching constants of triplet states of the dye by molecular oxygen, have been calculated. Luminescence quantum yield 1Δ g of polyvinylbutyral has been estimated. We have analyzed quantum-chemically electronic mechanisms of singlet-triplet annihilation of oxygen and eosin.
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LENUS (Irish Health Repository)
Limaye, Sandhya
2009-04-15
Flow cytometric techniques are increasingly used in pretransplant crossmatching, although there remains debate regarding the clinical significance and predictive value of donor-specific antibodies detected by flow cytometry. At least some of the discrepancies between published studies may arise from differences in cutoffs used and lack of standardization of the test.
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Derayea, Sayed M; Omar, Mahmoud A; Abu-Hassan, Ahmed A
2018-03-01
A simple, sensitive and non-extractive spectrofluorimetric method has been developed and validated for the determination of two psychoanaleptic drugs, atomoxetine and fluvoxamine, in pure forms and pharmaceutical dosage forms. The proposed method is based on the formation of binary complexes between eosin Y and the studied drugs in the presence of a Teorell-Stenhagen buffer. The quenching of the native fluorescence of eosin Y due to complex formation with the studied drugs was measured spectrofluorimetrically at 545 nm after excitation at 302 nm. At the optimum reaction conditions, the fluorescence quenching values (Δ F ) and concentrations were rectilinear over the concentration ranges of 0.2-2.2 and 0.3-2.2 µg ml -1 for atomoxetine and fluvoxamine, respectively. The developed method was successfully applied for the determination of the studied drugs in their pharmaceutical formulations with average percentage recoveries of 100.13 ± 0.66 and 99.69 ± 0.44 for atomoxetine and fluvoxamine, respectively ( n = 5), without interference from common excipients.
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Rosenauer, M.; Buchegger, W.; Finoulst, I.; Verhaert, P.D.E.M.; Vellekoop, M.
2010-01-01
In this study, the design, realization and measurement results of a novel optofluidic system capable of performing absorbance-based flow cytometric analysis is presented. This miniaturized laboratory platform, fabricated using SU-8 on a silicon substrate, comprises integrated polymer-based
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Liegler, T J; Hyun, W; Yen, T S; Stites, D P
1995-05-01
Regulation of peripheral lymphocyte number involves a poorly understood balance between cell renewal and loss. Disrupting this balance leads to a large number of disease states. Methods which allow qualitative and quantitative measurements of cell viability are increasingly valuable to studies directed at revealing the mechanisms underlying apoptotic and necrotic cell death. Here, we have characterized a method using single-laser flow cytometry that differentiates and quantifies the relative number of live, apoptotic, and late-stage apoptotic and necrotic peripheral lymphocytes. Following in vitro gamma irradiation and staining with acridine orange in combination with ethidium bromide, three distinct populations were seen by bivariate analysis of green versus red fluorescence. The identity of each distinct fluorescent population (whether live, apoptotic, or necrotic) was determined by sorting and examination of cellular morphology by electron microscopy. This flow cytometric method is directly compared with the techniques of trypan blue exclusion and DNA fragmentation to quantify cell death following exposure to various doses of in vitro gamma irradiation and postirradiation incubation times. We extend our findings to illustrate the utility of this method beyond analyzing radiation-induced apoptotic peripheral blood mononuclear cells (PBMC); similar fluorescent patterns are shown for radiation- and corticosteroid-treated murine thymocytes, activated human PBMC, and PBMC from human immunodeficiency virus-infected individuals. Our results demonstrate that dual-parameter flow cytometric analysis of acridine orange-ethidium bromide-stained lymphocytes is overall a superior method with increased sensitivity, greater accuracy, and decreased subjectivity in comparison with the other methods tested. By using standard laser and filter settings commonly available to flow cytometric laboratories, this method allows rapid measurement of a large number of cells from a
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Energy Technology Data Exchange (ETDEWEB)
Doering, U.
1997-02-01
To predict the movement of a contaminant plume in an aquifer is still a task of great uncertainty. This uncertainty is generally attributed to an insufficient understanding of the chemical reaction processes and/or to the natural aquifer heterogeneities. In an integrated approach of field experiments, laboratory experiments and numerical simulations, the transport of the weakly reactive solutes eosin, uranin and lithium was investigated at a test site near the Research Center in Juelich. The field scale transport behavior of the solutes was studied by large scale tracer tests. To characterize aquifer heterogeneities, in-situ and laboratory measurements were performed. In-situ measurements covered about 1500 flowmeter measurements and 90 determinations of the groundwater flow velocity by the borehole method. The spatial variability of hydraulic and physico-chemical parameters was further determined on 400 sediment samples. These parameters included: Grain size distribution, calculated hydraulic conductivity, unconformity and as physico-chemical parameters the organic carbon content, specific surface and the cation exchange capacity. Furthermore sorption coefficients were measured on 75 sediment samples for uranium and lithium. The statistical evaluation of these data showed that the hydraulic heterogeneity was larger but in the same order of magnitude as the physico-chemical parameters. (orig./SR) [Deutsch] Eine Schadstoff-Ausbreitung im Grundwasser vorherzusagen, ist noch immer eine Aufgabe mit unsicherem Ergebnis. Diese Prognose-Unsicherheiten werden im Allgemeinen auf ein unzureichendes Verstaendnis der chemischen Reaktionsprozesse und/oder auf die natuerliche Heterogenitaet des Grundwasserleiters zurueckgefuehrt. In dem hier beschriebenen Forschungsprojekt, das Feldversuche, Laborversuche und numerische Simulationen integriert, wurde der Transport der schwach reaktiven Substanzen Eosin, Uranin und Lithium auf einem Versuchsgelaende nahe des Forschungszentrums
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Flow cytometric measurement of DNA level and steroid hormone receptor assay in breast cancer
International Nuclear Information System (INIS)
Zubrikhina, G.N.; Kuz'mina, Eh.V.; Bassalyk, L.S.; Murav'eva, N.I.
1989-01-01
DNA level measured by flow cytometry and estrogen and progesteron receptors assayed in tissue samples obtained from 85 malignant and 16 benign lesions of the breast. All the benign tumors revealed 2c DNA content and most of them were receptor-negative, while 74.1% of breast carcinomas displayed aneuploidy. Three patients (3.5%) had two lines of aneuploid cells. Many aneuploid tumors were receptor-negative. Preoperative radiation treatmet (14-20 Gy) did not significantly influence the level of steroid hormone receptors in tumors. Estrogen receptor level was higher in menopausal patients than in premenopausal ones
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Directory of Open Access Journals (Sweden)
William Wilson Anku
2016-08-01
Full Text Available This paper reports the degradation of indigo carmine and eosin Y dyes in water, catalyzed by cobalt and multiwalled carbon nanotube modified zirconium oxide nanocomposite (Co-ZrO2-MWCNTs under simulated visible light. The bare ZrO2, ZrO2-MWCNTs, Co-ZrO2 and Co-ZrO2-MWCNTs with different percentage compositions of cobalt were synthesized by homogeneous co-precipitation method. Characterization of the prepared nanocomposites was carried out using X-Ray powder Diffraction (XRD, Fourier Transformer Infrared (FTIR Spectroscopy, Transmission Electron Microscopy (TEM, Raman Spectroscopy, (UV–Vis-Spectroscopy and Energy Dispersive Spectroscopy (EDS for their structure, formation, morphology, size and elemental analysis. The experimental results indicated that all the cobalt and MWCNTs modified nanocomposites demonstrated higher photocatalytic activities compared to the bare ZrO2. The most efficient catalyst (0.5% Co-ZrO2-MWCNTs with the band gap and Ka values of 5.21 eV and 16.86×10−3 min−1 respectively exhibited 98% degradation efficiency toward indigo carmine and 87% toward eosin Y in 180 min.
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LÜ, XINQUAN; DENG, QING; LI, HUIXIANG; SUO, ZHENHE
2011-01-01
Drug resistance of cancer stem/initiating cells has been considered to be one of the main reasons for tumor relapse. However, knowledge concerning the changes in stem/ initiating cells during chemotherapy is limited. In the present study, the breast cancer cell line MDA-MB-468 was cultured with 5-fluorouracil and serially passaged. Six cell generations were collected. Semi-quantitative RT-PCR and flow cytometric techniques were used to evaluate the protein and mRNA expression of stem/initiati...
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CSIR Research Space (South Africa)
Debnath, S
2015-01-01
Full Text Available Polyaniline-coated lignin-based adsorbent (PLC) was synthesized and used for uptake of reactive dye eosin yellow (EY) from aqueous solution. The adsorption capability of the adsorbent was found to be more effective than the unmodified adsorbent (LC...
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Wang, Peng; Zong, Lanlan; Guan, Zhongjie; Li, Qiuye; Yang, Jianjun
2018-02-01
An economic and effective Pt-based alloy cocatalyst has attracted considerable attention due to their excellent catalytic activity and reducing Pt usage. In this study, PtNi alloy cocatalyst was successfully decorated on the g-C3N4/GO hybrid photocatalyst via a facile chemical reduction method. The Eosin Y-sensitized g-C3N4/PtNi/GO-0.5% composite photocatalyst yields about 1.54 and 1178 times higher hydrogen evolution rate than the Eosin Y-sensitized g-C3N4/Pt/GO-0.5% and g-C3N4/Ni/GO-0.5% samples, respectively. Mechanism of enhanced performance for the g-C3N4/PtNi/GO composite was also investigated by different characterization, such as photoluminescence, transient photocurrent response, and TEM. These results indicated that enhanced charge separation efficiency and more reactive sites are responsible for the improved hydrogen evolution performance due to the positive synergetic effect between Pt and Ni. This study suggests that PtNi alloy can be used as an economic and effective cocatalyst for hydrogen evolution reaction. [Figure not available: see fulltext.
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Identification of ataxia telangiectasia heterozygotes by flow cytometric analysis of X-ray damage
International Nuclear Information System (INIS)
Rudolph, N.S.
1989-01-01
Flow cytometry was used to identify heterozygotes for the autosomal recessive DNA-repair deficiency disease ataxia telangiectasia (AT). Confluent G 0 /G 1 fibroblasts from 4 homozygotes (at/at), 5 obligate heterozygates (at/+) and 7 presumed normal (+/+) were X-irradiated with 200 Rad and subcultured immediately in medium containing 5-bromodeoxyuridine (BrdU). Cells were harvested 72 h later and stained with fluoresceinated anti-BrdU antibody to identify cells that had entered S phase. They were counterstained with propidium iodide to measure total DNA content. On the basis of relative release from G 0 /G 1 , the at/+ strains as a group were distinguished from both the presumed +/+ strains and at/at strains, although the individual values for some strains did show overlap between genotypes. When 10 cell strains were coded and analyzed in 'blind' experiments, all 4 heterozygotes were correctly assigned. By a similar assay in which exponentially growing cultures were pulsed briefly with BrdU 8 h after irradiation with 400 Rad and then harvested immediately, presumed +/+ cells as a group could be distinguished from at/at cells but not from at/- cells. This combination of assays assists in the identification of all 3 AT genotypes. This should be of both basic and diagnostic use, particularly in families known to segregate AT. (author). 37 refs.; 3 figs.; 5 tabs
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Energy Technology Data Exchange (ETDEWEB)
Chen, Hung-Hsia; Anbarasan, R.; Kuo, Long-Sheng [MEMS Thermal Control Lab, Department of Mechanical Engineering, National Taiwan University, Taipei 10617, Taiwan (China); Chen, Ping-Hei, E-mail: phchen@ntu.edu.tw [MEMS Thermal Control Lab, Department of Mechanical Engineering, National Taiwan University, Taipei 10617, Taiwan (China)
2011-04-15
Research highlights: {yields} In the present investigation, first, we functionalized the MWCNT with Eosin Y dye, for the ring opening polymerization. {yields} The dye functionalized MWCNT acted as an effective initiator which was further confirmed by various analytical techniques. {yields} By this methodology we saved the number of reaction steps, reaction time and also an economically cheaper one. - Abstract: This research conducted ring opening polymerization (ROP) of {epsilon}-caprolactone (C.L.) with a novel initiator, namely; Eosin Y functionalized multiwalled carbon nanotube (MWCNT) at 140 deg. C with nitrogen sparge under different concentrations. ROP of C.L. carried out at two different experimental conditions similar to variations in [M{sub 0}/I{sub 0}] and [C.L.]. Fourier transform infrared (FTIR) spectroscopy and nuclear magnetic resonance (NMR) spectroscopy confirmed the structure of MWCNT-Eosin Y initiated ROP of C.L. Gel permeation chromatography (GPC), X-ray photoelectron spectroscopy (XPS), UV-visible spectroscopy (UV-vis), scanning electron microscopy (SEM), and transmission electron microscopy (TEM) further characterized the structure. UV-visible spectroscopy determined the binding constant (K), for samples prepared under different [C.L.].
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International Nuclear Information System (INIS)
Chen, Hung-Hsia; Anbarasan, R.; Kuo, Long-Sheng; Chen, Ping-Hei
2011-01-01
Research highlights: → In the present investigation, first, we functionalized the MWCNT with Eosin Y dye, for the ring opening polymerization. → The dye functionalized MWCNT acted as an effective initiator which was further confirmed by various analytical techniques. → By this methodology we saved the number of reaction steps, reaction time and also an economically cheaper one. - Abstract: This research conducted ring opening polymerization (ROP) of ε-caprolactone (C.L.) with a novel initiator, namely; Eosin Y functionalized multiwalled carbon nanotube (MWCNT) at 140 deg. C with nitrogen sparge under different concentrations. ROP of C.L. carried out at two different experimental conditions similar to variations in [M 0 /I 0 ] and [C.L.]. Fourier transform infrared (FTIR) spectroscopy and nuclear magnetic resonance (NMR) spectroscopy confirmed the structure of MWCNT-Eosin Y initiated ROP of C.L. Gel permeation chromatography (GPC), X-ray photoelectron spectroscopy (XPS), UV-visible spectroscopy (UV-vis), scanning electron microscopy (SEM), and transmission electron microscopy (TEM) further characterized the structure. UV-visible spectroscopy determined the binding constant (K), for samples prepared under different [C.L.].
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Bui, Duc-Nguyen; Kang, Shi-Zhao; Qin, Lixia; Li, Xiang-Qing; Mu, Jin
2013-02-01
The photocatalytic system containing Eosin Y, multiwalled carbon nanotubes (MWNTs) and CuO (Eosin Y-MWNTs-CuO) was fabricated; meanwhile its photocatalytic activity for hydrogen evolution from triethanolamine (TEOA) aqueous solution was evaluated. Under visible light irradiation, the amount of hydrogen (H2) evolution increased greatly due to introduction of CuO in the photocatalytic system. Moreover, the electrochemical behavior of MWNTs loaded with CuO was explored using cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). The results clearly indicate that there is a strong relationship between the electrochemical behavior of MWNTs-CuO and the photocatalytic activity of Eosin Y-MWNTs-CuO, and the high photocatalytic activity of Eosin Y-MWNTs-CuO may mainly originate from the efficient electron-transfer in the system.
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Hashim, Noshin; Thakur, Shaila; Patang, Mouska; Crapulli, Ferdinando; Ray, Ajay K
2017-04-01
Diclofenac (DCF), a widely used non-steroidal anti-inflammatory drug, is a commonly detected substance that readily accumulates in tissues of aquatic fish and poses a threat to wildlife and freshwater quality. Advanced Oxidation Processes have been employed as an alternative due to the inadequacy of conventional treatment methods of trace contaminants. This study utilized an innovative method of solar-activation of TiO 2 using Eosin-Y dye for the degradation of DCF. Furthermore, the study incorporated a central composite design (CCD) to optimize the dye concentration and estimated the cost for the present process. Optimized parameters for light intensity (750 mW/cm 2 ), Eosin-Y dye concentration (2 mg/L), TiO 2 loading (37.5 mg/cm 2 ) and DCF concentration (25 mg/L) were determined through a CCD. The optimized parameters convey a DCF degradation rate of 40% and 49% for 2 ppm (low range) and 4 ppm (high range) dye concentrations, respectively, for a 5-minute reaction time. Cost estimation for the materials used was for the current process was also performed. It was determined that the additional cost of using 4 ppm instead of 2 ppm to achieve only 10% more DCF degradation is not warranted and would require additional treatment to remove subsequently formed halogenated compounds.
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Eosin Y Catalyzed Visible-Light-Promoted Aerobic Oxidative Cyclization of 2-Aminobenzothiazole
Directory of Open Access Journals (Sweden)
Vishal Srivastava
2015-12-01
Full Text Available A mild and efficient one-pot visible light irradiated synthesis of 2-aminobenzothiazole 4(a–l from arylisothiocyanate 1(a–l and secondary amines 2 have been reported in presence of eosin Y as an organophotoredox catalyst at room temperature under aerobic condition. This synthesis includes application of air and visible light as inexpensive, readily available, high atom economy, non-toxic and sustainable regents.
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Measurement of Soluble Biomarkers by Flow Cytometry
Antal-Szalm?s, P?ter; Nagy, B?la; Debreceni, Ildik? Beke; Kappelmayer, J?nos
2013-01-01
Microparticle based flow cytometric assays for determination of the level of soluble biomarkers are widely used in several research applications and in some diagnostic setups. The major advantages of these multiplex systems are that they can measure a large number of analytes (up to 500) at the same time reducing assay time, costs and sample volume. Most of these assays are based on antigen-antibody interactions and work as traditional immunoassays, but nucleic acid alterations ? by using spe...
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Shi, Jinwen; Guan, Xiangjiu; Zhou, Zhaohui; Liu, Haipei; Guo, Liejin
2015-06-01
Nanosheet (with around 20 nm in thickness)-stacked hollow-sphere TiO2 was synthesized via a modified solvothermal reaction for different times followed by calcination treatment at different temperatures. After surface modification by different cations (H+ or Fe3+) and further sensitization by Eosin Y, the obtained photocatalysts achieved remarkably enhanced H2-production activity (about 4.2 times of that for Eosin Y-sensitized P25) and stability under visible-light irradiation. The improved photocatalytic performance was synergistically caused by the enhanced Eosin Y sensitization (due to the enlarged surface area and electropositively modified surface), the optimized crystal structure (well-crystallized anatase phase), and the unique micro/nanostructure (nanosheet-stacked hollow spheres). This work presented an effective route to explore new visible-light-driven H2-production photocatalysts by coupling nanomaterials with special morphologies and metal-free dyes with visible-light absorption.
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1994-01-01
A simple method is described for high-resolution light and electron microscopic immunolocalization of proteins in cells and tissues by immunofluorescence and subsequent photooxidation of diaminobenzidine tetrahydrochloride into an insoluble osmiophilic polymer. By using eosin as the fluorescent marker, a substantial improvement in sensitivity is achieved in the photooxidation process over other conventional fluorescent compounds. The technique allows for precise correlative immunolocalization studies on the same sample using fluorescence, transmitted light and electron microscopy. Furthermore, because eosin is smaller in size than other conventional markers, this method results in improved penetration of labeling reagents compared to gold or enzyme based procedures. The improved penetration allows for three-dimensional immunolocalization using high voltage electron microscopy. Fluorescence photooxidation can also be used for high resolution light and electron microscopic localization of specific nucleic acid sequences by in situ hybridization utilizing biotinylated probes followed by an eosin-streptavidin conjugate. PMID:7519623
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International Nuclear Information System (INIS)
Shi, Jinwen; Guan, Xiangjiu; Zhou, Zhaohui; Liu, Haipei; Guo, Liejin
2015-01-01
Nanosheet (with around 20 nm in thickness)-stacked hollow-sphere TiO 2 was synthesized via a modified solvothermal reaction for different times followed by calcination treatment at different temperatures. After surface modification by different cations (H + or Fe 3+ ) and further sensitization by Eosin Y, the obtained photocatalysts achieved remarkably enhanced H 2 -production activity (about 4.2 times of that for Eosin Y-sensitized P25) and stability under visible-light irradiation. The improved photocatalytic performance was synergistically caused by the enhanced Eosin Y sensitization (due to the enlarged surface area and electropositively modified surface), the optimized crystal structure (well-crystallized anatase phase), and the unique micro/nanostructure (nanosheet-stacked hollow spheres). This work presented an effective route to explore new visible-light-driven H 2 -production photocatalysts by coupling nanomaterials with special morphologies and metal-free dyes with visible-light absorption
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Coexpression of multidrug resistance involve proteins: a flow cytometric analysis.
Boutonnat, J; Bonnefoix, T; Mousseau, M; Seigneurin, D; Ronot, X
1998-01-01
Cross resistance to multiple natural cytotoxic products represents a major obstacle in myeloblastic acute leukaemia (AML). Multidrug resistance (MDR) often involves overexpression of plasma membrane drug transporter P-glycoprotein (PGP) or the resistance associated protein (MRP). Recently, a protein overexpressed in a non-PGP MDR lung cancer cell line and termed lung resistance related protein (LRP) was identified. These proteins are known to be associated with a bad prognosis in AML. We have developed a triple indirect labelling analysed by flow cytometry to detect the coexpression of these proteins. Since no cell line expressing all three antigens is known, we mixed K562 cells (resistant to Adriblastine, PGP+, MRP-, LRP-) with GLC4 cells (resistant to Adriblastine, PGP-, MRP+, LRP+) to create a model system to test the method. The antibodies used were UIC2 for PGP, MRPm6 for MRP and LRP56 for LRP. They were revealed by Fab'2 coupled with Fluoresceine-isothiocyanate, Phycoerythrin or Tricolor with isotype specificity. Cells were fixed and permeabilized after PGP labelling because MRPm6 and LRP56 recognize intracellular epitopes. PGP and LRP were easily detected. MRP is expressed at relatively low levels and was more difficult to detect because in the triple labelling the non specific staining was higher than in a single labelling. Despite the increased background in the triple labelling we were able to detect coexpression of PGP, MRP, LRP by flow cytometry. This method appears to be very useful to detect coexpression of markers in AML. Such coexpression could modify the therapeutic approach with revertants.
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Directory of Open Access Journals (Sweden)
Wiltrud Haaß
Full Text Available ESPL1/Separase, an endopeptidase, is required for centrosome duplication and separation of sister-chromatides in anaphase of mitosis. Overexpression and deregulated proteolytic activity of Separase as frequently observed in human cancers is associated with the occurrence of supernumerary centrosomes, chromosomal missegregation and aneuploidy. Recently, we have hypothesized that increased Separase proteolytic activity in a small subpopulation of tumor cells may serve as driver of tumor heterogeneity and clonal evolution in chronic myeloid leukemia (CML. Currently, there is no quantitative assay to measure Separase activity levels in single cells. Therefore, we have designed a flow cytometry-based assay that utilizes a Cy5- and rhodamine 110 (Rh110-biconjugated Rad21 cleavage site peptide ([Cy5-D-R-E-I-M-R]2-Rh110 as smart probe and intracellular substrate for detection of Separase enzyme activity in living cells. As measured by Cy5 fluorescence the cellular uptake of the fluorogenic peptide was fast and reached saturation after 210 min of incubation in human histiocytic lymphoma U937 cells. Separase activity was recorded as the intensity of Rh110 fluorescence released after intracellular peptide cleavage providing a linear signal gain within a 90-180 min time slot. Compared to conventional cell extract-based methods the flow cytometric assay delivers equivalent results but is more reliable, bypasses the problem of vague loading controls and unspecific proteolysis associated with whole cell extracts. Especially suited for the investigaton of blood- and bone marrow-derived hematopoietic cells the flow cytometric Separase assay allows generation of Separase activity profiles that tell about the number of Separase positive cells within a sample i.e. cells that currently progress through mitosis and about the range of intercellular variation in Separase activity levels within a cell population. The assay was used to quantify Separase proteolytic
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Application of tot’hema eosin sensitized gelatin film for adaptive microlenses
Directory of Open Access Journals (Sweden)
Murić Branka D.
2017-01-01
Full Text Available In this paper we showed that tot’hema eosin sensitized gelatin (TESG film can be used for adaptive microlenses fabriacation. The mechanical properties of a pure gelatin film were improved by adding tot’hema solution. We found that the elasticity of TESG film depend on the tot’hema concentration. By stretching the film, the microlenses were deformed uniaxially, and microlenses focal length can be tuned. The achieved microlenses focal lengths range from 0.05 to 0.2 mm.
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Grilley-Olson, Juneko E.; Hayes, D. Neil; Moore, Dominic T.; Leslie, Kevin O.; Wilkerson, Matthew D.; Qaqish, Bahjat F.; Hayward, Michele C.; Cabanski, Christopher R.; Yin, Xiaoying; Socinski, Mark A.; Stinchcombe, Thomas E.; Thorne, Leigh B.; Allen, Timothy Craig; Banks, Peter M.; Beasley, Mary B.; Borczuk, Alain C.; Cagle, Philip T.; Christensen, Rebecca; Colby, Thomas V.; Deblois, Georgean G.; Elmberger, Göran; Graziano, Paolo; Hart, Craig F.; Jones, Kirk D.; Maia, Diane M.; Miller, C. Ryan; Nance, Keith V.; Travis, William D.; Funkhouser, William K.
2018-01-01
Context Precise subtype diagnosis of non–small cell lung carcinoma is increasingly relevant, based on the availability of subtype-specific therapies, such as bevacizumab and pemetrexed, and based on the subtype-specific prevalence of activating epidermal growth factor receptor mutations. Objectives To establish a baseline measure of inter-observer reproducibility for non–small cell lung carcinoma diagnoses with hematoxylin-eosin for the current 2004 World Health Organization classification, to estimate interobserver reproducibility for the therapeutically relevant squamous/nonsquamous subsets, and to examine characteristics that improve interobserver reproducibility. Design Primary, resected lung cancer specimens were converted to digital (virtual) slides. Based on a single hematoxylin-eosin virtual slide, pathologists were asked to assign a diagnosis using the 2004 World Health Organization classification. Kappa statistics were calculated for each pathologist-pair for each slide and were summarized by classification scheme, pulmonary pathology expertise, diagnostic confidence, and neoplastic grade. Results The 12 pulmonary pathology experts and the 12 community pathologists each independently diagnosed 48 to 96 single hematoxylin-eosin digital slides derived from 96 cases of non–small cell lung carcinoma resection. Overall agreement improved with simplification from the comprehensive 44 World Health Organization diagnoses (κ = 0.25) to their 10 major header subtypes (κ = 0.48) and improved again with simplification into the therapeutically relevant squamous/nonsquamous dichotomy (κ = 0.55). Multivariate analysis showed that higher diagnostic agreement was associated with better differentiation, better slide quality, higher diagnostic confidence, similar years of pathology experience, and pulmonary pathology expertise. Conclusions These data define the baseline diagnostic agreement for hematoxylin-eosin diagnosis of non–small cell lung carcinoma
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Energy Technology Data Exchange (ETDEWEB)
Zhang, Xiaojie [State Key Laboratory for Oxo Synthesis and Selective Oxidation, Lanzhou Institute of Chemical Physics, The Chinese Academy of Sciences, Lanzhou 730000 (China); Graduate University of the Chinese Academy of Sciences, Beijing 100101 (China); Jin, Zhiliang; Li, Shuben; Lu, Gongxuan [State Key Laboratory for Oxo Synthesis and Selective Oxidation, Lanzhou Institute of Chemical Physics, The Chinese Academy of Sciences, Lanzhou 730000 (China); Li, Yuexiang [Department of Chemistry, Nanchang University, Nanjing Road 245, Nanchang, 330047 (China)
2007-03-30
A new system for the production of hydrogen, constructed using silica gel as a matrix, Eosin Y as a photosensitizer, and Pt as a cocatalyst, has been reported. It was found that the rate of photosensitized hydrogen evolution in the presence of silica gel is enhanced about 10-fold relative to the homogeneous phase, i.e. in the absence of silica gel. The pH value of the solution and the concentration of Eosin Y have remarkable effects on the rate of hydrogen evolution. The optimal pH and concentration of Eosin Y are 7 and 3.60 x 10{sup -4} mol dm{sup -3} (E/S = 1/3) to 7.24 x 10{sup -4} mol dm{sup -3} (E/S = 1/1), respectively. Triethanolamine (TEOA) as an electron donor, the rate of hydrogen evolution and the apparent quantum efficiency in the silica gel system under visible-light irradiation ({lambda} {>=} 420 nm) can reach about 43 {mu}mol h{sup -1} and 10.4%, respectively. In addition, the roles of silica gel, Pt and TEOA, respectively; and the probable mechanism of photosensitized hydrogen evolution have been discussed. (author)
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Zhang, Xiaojie; Jin, Zhiliang; Li, Yuexiang; Li, Shuben; Lu, Gongxuan
A new system for the production of hydrogen, constructed using silica gel as a matrix, Eosin Y as a photosensitizer, and Pt as a cocatalyst, has been reported. It was found that the rate of photosensitized hydrogen evolution in the presence of silica gel is enhanced about 10-fold relative to the homogeneous phase, i.e. in the absence of silica gel. The pH value of the solution and the concentration of Eosin Y have remarkable effects on the rate of hydrogen evolution. The optimal pH and concentration of Eosin Y are 7 and 3.60 × 10 -4 mol dm -3 (E/S = 1/3) to 7.24 × 10 -4 mol dm -3 (E/S = 1/1), respectively. Triethanolamine (TEOA) as an electron donor, the rate of hydrogen evolution and the apparent quantum efficiency in the silica gel system under visible-light irradiation (λ ≥ 420 nm) can reach about 43 μmol h -1 and 10.4%, respectively. In addition, the roles of silica gel, Pt and TEOA, respectively; and the probable mechanism of photosensitized hydrogen evolution have been discussed.
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Detection of bacteriophage-infected cells of Lactococcus lactis using flow cytometry
DEFF Research Database (Denmark)
Michelsen, Ole; Cuesta-Dominguez, Álvaro; Albrektsen, Bjarne
2007-01-01
Bacteriophage infection in dairy fermentation constitutes a serious problem worldwide. We have studied bacteriophage infection in Lactococcus lactis by using the flow cytometer. The first effect of the infection of the bacterium is a change from cells in chains toward single cells. We interpret...... describe a new method for detection of phage infection in Lactococcus lactis dairy cultures. The method is based on flow cytometric detection of cells with low-density cell walls. The method allows fast and early detection of phage-infected bacteria, independently of which phage has infected the culture...
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Naumova, E. V.; Melnikov, A. G.; Melnikov, G. V.
2013-05-01
Fluorescence and phosphorescence quenching processes of polar and non-polar luminescent probes associated with human serum albumin (HSA) in phosphate buffer at pH 7.4 were studied. Stern-Volmer quenching constants of anthracene and pyrene fluorescence and eosin phosphorescence and rate constants for quenching of eosin triplet states were determined. The polarity index of pyrene bound to HSA was obtained as a function of thallium nitrate concentration. The influences of structural changes in the proteins that were stimulated by heavy-metal salts and of screening of protein charges by salt ions on quenching processes of singlet and triplet states of the probes were found.
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2′-deoxy-5,6-dihydro-5-azacytidine—a less toxic alternative of 2′-deoxy-5-azacytidine
Matoušová, Marika; Votruba, Ivan; Otmar, Miroslav; Tloušťová, Eva; Günterová, Jana
2011-01-01
Restoration of transcriptionally silenced genes by means of methyltransferases inhibitors plays a crucial role in the current therapy of myelodysplastic syndromes and certain types of leukemias. A comparative study of hypomethylating activities of a series of 5-azacytidine nucleosides: 5-azacytidine (AC), 2′-deoxy-5-azacytidine (DAC) and its α-anomer (α-DAC), 5,6-dihydro-5-azacytidine (DHAC), 2′-deoxy-5,6-dihydro-5-azacytidine (DHDAC, KP-1212) and its α-anomer (α-DHDAC), and of a 2-pyrimidone ribonucleoside (zebularine) was conducted. Methylation-specific PCR was employed to detect the efficiency of individual agents on cyclin-dependent kinase inhibitor 2B and thrombospondin-1 hypermethylated gene loci. Overall changes in DNA methylation level were quantified by direct estimation of 5-methyl-2′-deoxycytidine-5′-monophosphate by HPLC using digested genomic DNA. Flow cytometric analysis of cell cycle progression and apoptotic markers was used to determine cytotoxicity of the compounds. mRNA expression was measured using qRT-PCR. 2′-deoxy-5,6-dihydro-5-azacytidine was found to be less cytotoxic and more stable than 2′-deoxy-5-azacytidine at the doses that induce comparable DNA hypomethylation and gene reactivation. This makes it a valuable tool for epigenetic research and worth further investigations to elucidate its possible therapeutic potential. PMID:21566456
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Eosinófilos sanguíneos e fecais em crianças infectadas por helmintos e protozoários
Silva, Vanessa Marques de Araújo
2012-01-01
Em infecções parasitárias helmínticas a eosinofilia é um dos fatores essenciais à resistência do hospedeiro, entretanto, infecções com protozoários raramente resultam em eosinofilia periférica. O objetivo deste estudo foi identificar infecções por enteroparasitas em crianças e associá-las com eosinófilos sanguíneos e fecais. Os exames parasitológicos efetuados pelo método de Blagg, a pesquisa de eosinófilos pelo tricômio e coloração por Leishman foram realizados em 346 crianças. Amostras posi...
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The utility of flow sorting to identify chromosomes carrying a single copy transgene in wheat
Czech Academy of Sciences Publication Activity Database
Cápal, Petr; Endo, Takashi R.; Vrána, Jan; Kubaláková, Marie; Karafiátová, Miroslava; Komínková, Eva; Mora-Ramirez, I.; Weschke, W.; Doležel, Jaroslav
2016-01-01
Roč. 12, APR 25 (2016), s. 24 ISSN 1746-4811 R&D Projects: GA MŠk(CZ) LO1204; GA ČR GBP501/12/G090 Institutional support: RVO:61389030 Keywords : Transgene localization * Flow cytometric sorting * Single chromosome amplification Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 3.510, year: 2016
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Caires, Cynthia S A; Leal, Cassia R B; Ramos, Carlos A N; Bogo, Danielle; Lima, Alessandra R; Arruda, Eduardo J; Oliveira, Samuel L; Caires, Anderson R L; Nascimento, Valter A
2017-07-01
The use of eosin methylene blue according to Giemsa as photosensitizer is presented for the first time in this paper. The present study evaluated the potential application of chlorophyllin sodium copper salt (CuChlNa) and eosin methylene blue according to Giemsa (EMB) as antimicrobial photosensitizers (aPS) for photodynamic inactivation (PDI) of Staphylococcus aureus (gram-positive) and Escherichia coli (gram-negative) bacteria. The experiments were performed using S. aureus stain ATCC 25923 and E. coli ATCC 25922 in which five aPS concentrations (0.0, 1.0, 2.5, 5.0, 10.0, and 20.0 μM for S. aureus and 0.0, 5.0, 10.0, 20.0, 40.0, and 50.0 μM for E. coli) were prepared and added in 2 mL of a saline solution containing the bacterial inoculum. After aPS incubation, the samples were divided into two groups, one kept in the dark and another submitted to the illumination. Then, the bacterial inactivation was determined 18 h after the incubation at 37 °C by counting the colony-forming units (CFU). The results revealed that both EMB and CuChlNa can be used as aPS for the photoinactivation of S. aureus, while only EMB was able to photoinactivate E. coli. Nevertheless, a more complex experimental setup was needed for photoinactivation of E. coli. The data showed that EMB and CuChlNa presented similar photoinactivation effects on S. aureus, in which bacterial growth was completely inhibited at photosensitizer (PS) concentrations over 5 μM, when samples were previously incubated for 30 min and irradiated by a light dose of 30 J cm -2 as a result of an illumination of 1 h at 8.3 mW cm -2 by using a red light at 625 nm with a 1 cm beam diameter and output power of 6.5 mW. In the case of E. coli, bacterial growth was completely inhibited only when combining a PS incubation period of 120 min with concentrations over 20 μM.
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Energy Technology Data Exchange (ETDEWEB)
Sreethawong, Thammanoon; Chavadej, Sumaeth [The Petroleum and Petrochemical College, Chulalongkorn University, Soi Chula 12, Phyathai Road, Pathumwan, Bangkok 10330 (Thailand); Center for Petroleum, Petrochemicals, and Advanced Materials, Chulalongkorn University, Bangkok 10330 (Thailand); Junbua, Chompoonuch [The Petroleum and Petrochemical College, Chulalongkorn University, Soi Chula 12, Phyathai Road, Pathumwan, Bangkok 10330 (Thailand)
2009-05-15
Sensitized photocatalytic production of hydrogen from water splitting is investigated under visible light irradiation over mesoporous-assembled titanium dioxide (TiO{sub 2}) nanocrystal photocatalysts, without and with Pt loading. The photocatalysts are synthesized by a sol-gel process with the aid of a structure-directing surfactant and are characterized by N{sub 2} adsorption-desorption analysis, X-ray diffraction, UV-vis spectroscopy, scanning electron microscopy, transmission electron microscopy and energy-dispersive X-ray analysis. The dependence of hydrogen production on the type of TiO{sub 2} photocatalyst (synthesized mesoporous-assembled and commercial non-mesoporous-assembled TiO{sub 2} without and with Pt loading), the calcination temperature of the synthesized photocatalyst, the sensitizer (Eosin Y) concentration, the electron donor (diethanolamine) concentration, the photocatalyst dosage and the initial solution pH is systematically studied. The results show that in the presence of the Eosin Y sensitizer, the Pt-loaded mesoporous-assembled TiO{sub 2} synthesized by a single-step sol-gel process and calcined at 500 C exhibits the highest photocatalytic activity for hydrogen production from a 30 vol.% diethanolamine aqueous solution with dissolved 2 mM Eosin Y. Moreover, the optimum photocatalyst dosage and initial solution pH for the maximum photocatalytic activity for hydrogen production are 3.33 g dm{sup -3} and 11.5, respectively. (author)
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Matoušová, Marika; Votruba, Ivan; Otmar, Miroslav; Tloušťová, Eva; Günterová, Jana; Mertlíková-Kaiserová, Helena
2011-06-01
Restoration of transcriptionally silenced genes by means of methyltransferases inhibitors plays a crucial role in the current therapy of myelodysplastic syndromes and certain types of leukemias. A comparative study of hypomethylating activities of a series of 5-azacytidine nucleosides: 5-azacytidine (AC), 2'-deoxy-5-azacytidine (DAC) and its α-anomer (α-DAC), 5,6-dihydro-5-azacytidine (DHAC), 2'-deoxy-5,6-dihydro-5-azacytidine (DHDAC, KP-1212) and its α-anomer (α-DHDAC), and of a 2-pyrimidone ribonucleoside (zebularine) was conducted. Methylation-specific PCR was employed to detect the efficiency of individual agents on cyclin-dependent kinase inhibitor 2B and thrombospondin-1 hypermethylated gene loci. Overall changes in DNA methylation level were quantified by direct estimation of 5-methyl-2'-deoxycytidine-5'-monophosphate by HPLC using digested genomic DNA. Flow cytometric analysis of cell cycle progression and apoptotic markers was used to determine cytotoxicity of the compounds. mRNA expression was measured using qRT-PCR. 2'-deoxy-5,6-dihydro-5-azacytidine was found to be less cytotoxic and more stable than 2'-deoxy-5-azacytidine at the doses that induce comparable DNA hypomethylation and gene reactivation. This makes it a valuable tool for epigenetic research and worth further investigations to elucidate its possible therapeutic potential.
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Il Koo, Bon; Lee, Yun-Soo; Seo, Mintae; Seok Choi, Hyung; Leng Seah, Geok; Nam, Taegu; Nam, Yoon Sung
2017-07-31
Chemical biocides have been widely used as marine antifouling agents, but their environmental toxicity impose regulatory restriction on their use. Although various surrogate antifouling biocides have been introduced, their comparative effectiveness has not been well investigated partly due to the difficulty of quantitative evaluation of their antifouling activity. Here we report an image cytometric method to quantitatively analyze the antifouling activities of seven commercial biocides using Ulva prolifera as a target organism, which is known to be a dominant marine species causing soft fouling. The number of spores settled on a substrate is determined through image analysis using the intrinsic fluorescence of chlorophylls in the spores. Pre-determined sets of size and shape of spores allow for the precise determination of the number of settled spores. The effects of biocide concentration and combination of different biocides on the spore settlement are examined. No significant morphological changes of Ulva spores are observed, but the amount of adhesive pad materials is appreciably decreased in the presence of biocides. It is revealed that the growth rate of Ulva is not directly correlated with the antifouling activities against the settlement of Ulva spores. This work suggests that image cytometric analysis is a very convenient, fast-processable method to directly analyze the antifouling effects of biocides and coating materials.
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Walash, M. I.; Belal, F.; El-Enany, N.; Elmansi, H.
2010-01-01
Spectrophotometric and spectrofluorimetric methods were developed for the determination of dothiepin hydrochloride (DOP) in different dosage forms. The spectrophotometric method (Method I) is based on formation of a binary complex with eosin at 540 nm in acetate buffer of pH3.7. The absorbance-concentration plot is rectilinear over the range 1–10 μg/mL with LOD of 0.18 μg/mL and LOQ of 0.54 μg/mL. The spectroflurimetric method (Method II) is based on the quantitative quenching effect of Dothiepin on the native fluorescence of eosin at the same pH. The quenching of the fluorescence of eosin was measured at 543 nm after excitation at 304 nm. The fluorescence-concentration plot is rectilinear over the range 0.3–8 μg/ mL with LOD of 0.11 μg/mL and LOQ of 0.34 μg/mL. The proposed methods were successfully applied to the analysis of commercial tablets and capsules containing the drug. Statistical comparison of the results with those of the reference method revealed good agreement and proved that there were no significant differences in the accuracy and precision between the two methods respectively. PMID:23675210
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Microbial Eco-Physiology of the human intestinal tract: a flow cytometric approach
Amor, Ben K.
2004-01-01
This thesis describes a multifaceted approach to further enhance our view of the complex human intestinal microbial ecosystem. This approach combines me advantages of flow cyrometry (FCM), a single cell and high-throughput technology, and molecular techniques that have proven themselves to be
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International Nuclear Information System (INIS)
Shen Yan; Tefashe, Ushula Mengesha; Nonomura, Kazuteru; Loewenstein, Thomas; Schlettwein, Derck; Wittstock, Gunther
2009-01-01
The overall efficiency of the light-induced charge separation in dye-sensitized solar cells depends on the kinetic competition between back electron transfer and dye regeneration processes by a redox electrolyte. In a previous study, the reduction of the intermittently formed photo-oxidized dye molecules by iodide ions in the electrolyte phase was investigated using the feedback mode of a scanning electrochemical microscope (SECM) and a quantitative model had been derived. Here we provide a more thorough experimental verification of this model by variation of the excitation wavelength, light intensities and mediator concentrations. Nanoporous ZnO/Eosin Y films prepared by self-assembly were used as model electrodes and were used with an iodide/triiodide electrolyte. The experimentally found effective rate constants could be related to the rate constant for the reaction of the dissolved donor with photo-oxidized Eosin Y bound to ZnO and the absorption spectrum of the dye and confirmed the assumption made in the derivation of the model. For the regeneration process of Eosin Y, a rate constant of k ox with different light emitting diodes and light intensities is determined.
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Energy Technology Data Exchange (ETDEWEB)
Shen Yan; Tefashe, Ushula Mengesha [Department of Pure and Applied Chemistry, Faculty of Mathematics and Natural Sciences, Carl von Ossietzky University of Oldenburg, D-26111 Oldenburg (Germany); Nonomura, Kazuteru; Loewenstein, Thomas; Schlettwein, Derck [Institute of Applied Physics, Justus Liebig University of Giessen, Heinrich-Buff-Ring 16, D-35392 Giessen (Germany); Wittstock, Gunther, E-mail: gunther.wittstock@uni-oldenburg.d [Department of Pure and Applied Chemistry, Faculty of Mathematics and Natural Sciences, Carl von Ossietzky University of Oldenburg, D-26111 Oldenburg (Germany)
2009-12-30
The overall efficiency of the light-induced charge separation in dye-sensitized solar cells depends on the kinetic competition between back electron transfer and dye regeneration processes by a redox electrolyte. In a previous study, the reduction of the intermittently formed photo-oxidized dye molecules by iodide ions in the electrolyte phase was investigated using the feedback mode of a scanning electrochemical microscope (SECM) and a quantitative model had been derived. Here we provide a more thorough experimental verification of this model by variation of the excitation wavelength, light intensities and mediator concentrations. Nanoporous ZnO/Eosin Y films prepared by self-assembly were used as model electrodes and were used with an iodide/triiodide electrolyte. The experimentally found effective rate constants could be related to the rate constant for the reaction of the dissolved donor with photo-oxidized Eosin Y bound to ZnO and the absorption spectrum of the dye and confirmed the assumption made in the derivation of the model. For the regeneration process of Eosin Y, a rate constant of k{sub ox} with different light emitting diodes and light intensities is determined.
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Nagaya, Satoshi; Nishikiori, Hiromasa; Mizusaki, Hideaki; Wagata, Hajime; Teshima, Katsuya
2015-06-03
The thin films consisting of crystalline ZnO particles were prepared on fluorine-doped tin oxide electrodes by electroless deposition. The particles were deposited from an aqueous solution containing zinc nitrate, dimethyamine-borane, and eosin Y at 328 K. As the Pd particles were adsorbed on the substrate, not only the eosin Y monomer but also the dimer and debrominated species were rapidly adsorbed on the spherical ZnO particles, which were aggregated and formed secondary particles. On the other hand, in the absence of the Pd particles, the monomer was adsorbed on the flake-shaped ZnO particles, which vertically grew on the substrate surface and had a high crystallinity. The photoelectric conversion efficiency was higher for the ZnO electrodes containing a higher amount of the monomer during light irradiation.
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Energy Technology Data Exchange (ETDEWEB)
Ryan, M.A.; Fitzgerald, E.C.; Spitler, M.T. (Polaroid Corp., Cambridge, MA (USA))
1989-08-10
It is shown that the photoelectrochemical data on eosin Y sensitized TiO{sub 2} single-crystal electrodes cannot be interpreted unambiguously without concomitant data from flash photolysis measurements on this system. By use of a combination of internal reflection spectroscopy and laser flash photolysis, electron exchange with TiO{sub 2} was observed for the excited singlet state, the triplet state, and the cation radical of the dye. With a temporal resolution of 100 ns, the kinetics of the charge transfer are compared with those of the dye in solution and used to interpret the photoelectrochemistry of the dye at the electrode. Spectroscopic evidence revealed photocurrent production by the triplet state and a reduction of the eosin cation radical by electrons from the TiO{sub 2} conduction band and by hydroquinone.
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Energy Technology Data Exchange (ETDEWEB)
Shi, Jinwen, E-mail: jinwen-shi@mail.xjtu.edu.cn; Guan, Xiangjiu; Zhou, Zhaohui; Liu, Haipei; Guo, Liejin [Xi’an Jiaotong University (XJTU), State Key Laboratory of Multiphase Flow in Power Engineering (MFPE), International Research Center for Renewable Energy IRCRE (China)
2015-06-15
Nanosheet (with around 20 nm in thickness)-stacked hollow-sphere TiO{sub 2} was synthesized via a modified solvothermal reaction for different times followed by calcination treatment at different temperatures. After surface modification by different cations (H{sup +} or Fe{sup 3+}) and further sensitization by Eosin Y, the obtained photocatalysts achieved remarkably enhanced H{sub 2}-production activity (about 4.2 times of that for Eosin Y-sensitized P25) and stability under visible-light irradiation. The improved photocatalytic performance was synergistically caused by the enhanced Eosin Y sensitization (due to the enlarged surface area and electropositively modified surface), the optimized crystal structure (well-crystallized anatase phase), and the unique micro/nanostructure (nanosheet-stacked hollow spheres). This work presented an effective route to explore new visible-light-driven H{sub 2}-production photocatalysts by coupling nanomaterials with special morphologies and metal-free dyes with visible-light absorption.
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Synthesis of 1-((4-methoxyphenyl-3-alkynes-1H-pyrrole-2,5-diones and functionalization to tria
Directory of Open Access Journals (Sweden)
Bakhat Ali
2017-05-01
Full Text Available A series of alkynyl maleimides were prepared via one-step cross-coupling reaction using bromomaleimide and acetylenes under the Sonogashira conditions, affording 1-((4-methoxyphenyl-3-alkynes-1H-pyrrole-2,5-diones in good to high yields. These products were subsequently converted in the corresponding 1,2,3-triazole using conventional click chemistry approach. The alkynyl maleimide compound (8g crystallized in the triclinic space group P1 with unit cell parameters a = 5.3692(6, b = 9.2513(10, c = 10.3070(11 Å, α = 85.349(4, β = 86.892(4, γ = 86.892(4°, V = 507.31(10 Å3, and Z = 1. In the crystal the molecules are stacked parallel to the c axis and held together through a C–H⋯π and a C–H⋯O interaction.
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Directory of Open Access Journals (Sweden)
Eid Manal I
2011-10-01
Full Text Available Abstract A simple, sensitive and rapid spectrophotometric method was developed and validated for the determination of two skeletal muscle relaxants namely, tizanidine hydrochloride (I and orphenadrine citrate (II in pharmaceutical formulations. The proposed method is based on the formation of a binary complex between the studied drugs and eosin Y in aqueous buffered medium (pH 3.5. Under the optimum conditions, the binary complex showed absorption maxima at 545 nm for tizanidine and 542 nm for orphenadrine. The calibration plots were rectilinear over concentration range of 0.5-8 μg/mL and 1-12 μg/mL with limits of detection of 0.1 μg/mL and 0.3 μg/mL for tizanidine and orphenadrine respectively. The different experimental parameters affecting the development and stability of the complex were studied and optimized. The method was successfully applied for determination of the studied drugs in their dosage forms; and to the content uniformity test of tizanidine in tablets.
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Mustapha, Pascale; Epalle, Thibaut; Allegra, Séverine; Girardot, Françoise; Garraud, Olivier; Riffard, Serge
2015-04-01
The viability of three Legionella pneumophila strains was monitored after chlorine dioxide (ClO2) treatment using a flow cytometric assay. Suspensions of L. pneumophila cells were submitted to increasing concentrations of ClO2. Culturable cells were still detected when using 4 mg/L, but could no longer be detected after exposure to 6 mg/L of ClO2, although viable but not culturable (VBNC) cells were found after exposure to 4-5 mg/L of ClO2. When testing whether these VBNC were infective, two of the strains were resuscitated after co-culture with Acanthamoeba polyphaga, but neither of them could infect macrophage-like cells. Copyright © 2015 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.
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Size-dependent production of radicals in catalyzed reduction of Eosin Y using gold nanorods
Weng, Guojun; Qi, Ying; Li, Jianjun; Zhao, Junwu
2015-09-01
Gold nanostructures have been widely used as catalysts for chemical processes, energy conversion, and pollution control. The size of gold nanocatalysts is thus paramount for their catalytic activity. In this paper, gold nanorods with different sizes were prepared by means of the improved seeding growth approach by adding aromatic additive. The sizes and aspect ratios of the obtained gold nanorods were calculated according to the TEM characterization. Then, we studied the catalytic activities of gold nanorods using a model reaction based on the reduction of Eosin Y by NaBH4. By monitoring the absorption intensities of the radicals induced by gold nanorods in real time, we observed the clear size-dependent activity in the conversion of EY2- to EY3-. The conversion efficiency indicated that the gold nanorods with the smallest size were catalytically the most active probably due to their high number of coordinatively unsaturated surface atoms. In addition, a compensation effect dominated by the surface area of nanorods was observed in this catalytic reduction, which could be primarily attributed to the configuration of Eosin Y absorbed onto the surfaces of gold nanorods.
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Kłeczek, Paweł; Dyduch, Grzegorz; Jaworek-Korjakowska, Joanna; Tadeusiewicz, Ryszard
2017-03-01
Background: Epidermis area is an important observation area for the diagnosis of inflammatory skin diseases and skin cancers. Therefore, in order to develop a computer-aided diagnosis system, segmentation of the epidermis area is usually an essential, initial step. This study presents an automated and robust method for epidermis segmentation in whole slide histopathological images of human skin, stained with hematoxylin and eosin. Methods: The proposed method performs epidermis segmentation based on the information about shape and distribution of transparent regions in a slide image and information about distribution and concentration of hematoxylin and eosin stains. It utilizes domain-specific knowledge of morphometric and biochemical properties of skin tissue elements to segment the relevant histopathological structures in human skin. Results: Experimental results on 88 skin histopathological images from three different sources show that the proposed method segments the epidermis with a mean sensitivity of 87 %, a mean specificity of 95% and a mean precision of 57%. It is robust to inter- and intra-image variations in both staining and illumination, and makes no assumptions about the type of skin disorder. The proposed method provides a superior performance compared to the existing techniques.
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Size-dependent production of radicals in catalyzed reduction of Eosin Y using gold nanorods
Energy Technology Data Exchange (ETDEWEB)
Weng, Guojun; Qi, Ying; Li, Jianjun; Zhao, Junwu, E-mail: nanoptzhao@163.com [Xi’an Jiaotong University, The Key Laboratory of Biomedical Information Engineering of Ministry of Education, School of Life Science and Technology (China)
2015-09-15
Gold nanostructures have been widely used as catalysts for chemical processes, energy conversion, and pollution control. The size of gold nanocatalysts is thus paramount for their catalytic activity. In this paper, gold nanorods with different sizes were prepared by means of the improved seeding growth approach by adding aromatic additive. The sizes and aspect ratios of the obtained gold nanorods were calculated according to the TEM characterization. Then, we studied the catalytic activities of gold nanorods using a model reaction based on the reduction of Eosin Y by NaBH{sub 4}. By monitoring the absorption intensities of the radicals induced by gold nanorods in real time, we observed the clear size-dependent activity in the conversion of EY{sup 2−} to EY{sup 3−}. The conversion efficiency indicated that the gold nanorods with the smallest size were catalytically the most active probably due to their high number of coordinatively unsaturated surface atoms. In addition, a compensation effect dominated by the surface area of nanorods was observed in this catalytic reduction, which could be primarily attributed to the configuration of Eosin Y absorbed onto the surfaces of gold nanorods.
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Size-dependent production of radicals in catalyzed reduction of Eosin Y using gold nanorods
International Nuclear Information System (INIS)
Weng, Guojun; Qi, Ying; Li, Jianjun; Zhao, Junwu
2015-01-01
Gold nanostructures have been widely used as catalysts for chemical processes, energy conversion, and pollution control. The size of gold nanocatalysts is thus paramount for their catalytic activity. In this paper, gold nanorods with different sizes were prepared by means of the improved seeding growth approach by adding aromatic additive. The sizes and aspect ratios of the obtained gold nanorods were calculated according to the TEM characterization. Then, we studied the catalytic activities of gold nanorods using a model reaction based on the reduction of Eosin Y by NaBH 4 . By monitoring the absorption intensities of the radicals induced by gold nanorods in real time, we observed the clear size-dependent activity in the conversion of EY 2− to EY 3− . The conversion efficiency indicated that the gold nanorods with the smallest size were catalytically the most active probably due to their high number of coordinatively unsaturated surface atoms. In addition, a compensation effect dominated by the surface area of nanorods was observed in this catalytic reduction, which could be primarily attributed to the configuration of Eosin Y absorbed onto the surfaces of gold nanorods
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Andrés-Manzano, M Jesús; Andrés, Vicente; Dorado, Beatriz
2015-01-01
Methods for staining tissues with Oil Red O and hematoxylin-eosin are classical histological techniques that are widely used to quantify atherosclerotic burden in mouse tissues because of their ease of use, reliability, and the large amount of information they provide. These stains can provide quantitative data about the impact of a genetic or environmental factor on atherosclerotic burden and on the initiation, progression, or regression of the disease, and can also be used to evaluate the efficacy of drugs designed to prevent or treat atherosclerosis. This chapter provides protocols for quantifying atherosclerotic burden in mouse aorta and aortic root, including methods for dissection, Oil Red O staining, hematoxylin-eosin staining, and image analysis.
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International Nuclear Information System (INIS)
Cremer, C.; Cremer, T.; Gray, J.W.
1982-01-01
Asynchronously growing cells of a M3-1 Chinese hamster line were ultraviolet (UV) irradiated (lambda . 254 nm) with UV fluences up to 7.5 J/m(2). After irradiation cells were incubated with or without 2 mM caffeine for 20 hr, then mitotic cells were selected by mechanical shaking. Their chromosomes were isolated, stained with Hoechst 33258 and chromomycin A3, and measured flow cytometrically. While the fluorescence distributions of chromosomes (flow karyo-types) from cells treated with UV alone or with caffeine alone were very similar to those of untreated controls, the flow karyo-types of UV + caffeine-treated cells showed a debris continuum that increased with increasing UV fluence suggesting an increased number of chromosome fragments. Visual evaluation of metaphase plates revealed that the percentage of cells with chromosome damage also increased steadily with increasing UV fluence. A high degree of correlation was observed between the relative magnitude of the debris level from flow karyotypes and the percentage of cells with chromosome damage and with generalized chromosomes shattering, respectively, as determined from metaphase spreads
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Directory of Open Access Journals (Sweden)
Zhou F
2018-03-01
Full Text Available Fangbin Zhou,1,2 Yaying Zhou,3 Ming Yang,1 Jinli Wen,3 Jun Dong,4 Wenyong Tan1 1Department of Oncology, The Second Clinical Medical College, Shenzhen People’s Hospital, Jinan University, Shenzhen, People’s Republic of China; 2Integrated Chinese and Western Medicine Postdoctoral Research Station, Jinan University, Guangzhou, People’s Republic of China; 3Clinical Medical Research Center, The Second Clinical Medical College, Shenzhen People’s Hospital, Jinan University, Shenzhen, People’s Republic of China; 4Department of Pathophysiology, Key Laboratory of the State Administration of Traditional Chinese Medicine, Medical College of Jinan University, Guangzhou, People’s Republic of China Background: Circulating endothelial cells (CECs and their subpopulations could be potential novel biomarkers for various malignancies. However, reliable enumerable methods are warranted to further improve their clinical utility. This study aimed to optimize a flow cytometric method (FCM assay for CECs and subpopulations in peripheral blood for patients with solid cancers.Patients and methods: An FCM assay was used to detect and identify CECs. A panel of 60 blood samples, including 44 metastatic cancer patients and 16 healthy controls, were used in this study. Some key issues of CEC enumeration, including sample material and anticoagulant selection, optimal titration of antibodies, lysis/wash procedures of blood sample preparation, conditions of sample storage, sufficient cell events to enhance the signal, fluorescence-minus-one controls instead of isotype controls to reduce background noise, optimal selection of cell surface markers, and evaluating the reproducibility of our method, were integrated and investigated. Wilcoxon and Mann–Whitney U tests were used to determine statistically significant differences.Results: In this validation study, we refined a five-color FCM method to detect CECs and their subpopulations in peripheral blood of patients
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de Keijzer, M.; van Bommel, M.R.; Kirby, J.
2017-01-01
This paper discusses the history, chemical constitution, production, dyeing processes and properties of the hydroxyphthaleins. Important dyes of this dye class are fluorescein, the eosins, phloxines, erythrosines, rose bengals and rhodamines. These dyes were used for textile dyeing and as painting
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Rapid assay for cell age response to radiation by electronic volume flow cell sorting
International Nuclear Information System (INIS)
Freyer, J.P.; Wilder, M.E.; Raju, M.R.
1987-01-01
A new technique is described for measuring cell survival as a function of cell cycle position using flow cytometric cell sorting on the basis of electronic volume signals. Sorting of cells into different cell age compartments is demonstrated for three different cell lines commonly used in radiobiological research. Using flow cytometric DNA content analysis and [ 3 H]thymidine autoradiography of the sorted cell populations, it is demonstrated that resolution of the age compartment separation is as good as or better than that reported for other cell synchronizing techniques. Variation in cell survival as a function of position in the cell cycle after a single dose of radiation as measured by volume cell sorting is similar to that determined by other cell synchrony techniques. Advantages of this method include: (1) no treatment of the cells is required, thus, this method is noncytotoxic; (2) no cell cycle progression is needed to obtain different cell age compartments; (3) the cell population can be held in complete growth medium at any desired temperature during sorting; (4) a complete radiation age - response assay can be plated in 2 h. Applications of this method are discussed, along with some technical limitations. (author)
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A flow cytometric method for assessing viability of intraerythrocytic hemoparasites.
Wyatt, C R; Goff, W; Davis, W C
1991-06-24
We have developed a rapid, reliable method of evaluating growth and viability of intraerythrocytic protozoan hemoparasites. The assay involves the selective uptake and metabolic conversion of hydroethidine to ethidium by live parasites present in intact erythrocytes. The red fluorescence imparted by ethidium intercalated into the DNA of the parasite permits the use of flow cytometry to distinguish infected erythrocytes with viable parasites from uninfected erythrocytes and erythrocytes containing dead parasites. Comparison of the fluorochromasia technique of enumerating the number and viability of hemoparasites in cultured erythrocytes with enumeration in Giemsa-stained films and uptake of [3H]hypoxanthine demonstrated the fluorochromasia technique yields comparable results. Studies with the hemoparasite, Babesia bovis, have shown the fluorochromasia technique can also be used to monitor the effect of parasiticidal drugs on parasites in vitro. The cumulative studies with the fluorochromasia assay suggest the assay will also prove useful in investigations focused on analysis of the immune response to hemoparasites and growth in vitro.
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Directory of Open Access Journals (Sweden)
Mahasin F. Hadi Al-Kadhemy
2017-07-01
Full Text Available In this research, X-ray diffraction of the powder (PVP polymer, Eosin B dye, and silver nitrate and (EB/PVP, AgNO3/PVP, EB/AgNO3/PVP films have been studied. Casting method is used to prepare homogeneous films on plastic petri dishes. All parameters accounted for the X-ray diffraction; full width half maximum (FWHM, Miller indices (hkl, size of crystalline (D, Specific Surface Area (S and Dislocation Density (δ.The nature of the structural of materials and films will be investigated. The XRD pattern of PVP polymer was amorphous structure with two broader peaks and the Eosin B dye and silver nitrate have crystalline structure. While the mixture between these materials led to appearing some crystalline peaks into XRD pattern of PVP polymer.
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Glier, Hana; Heijnen, Ingmar; Hauwel, Mathieu; Dirks, Jan; Quarroz, Stéphane; Lehmann, Thomas; Rovo, Alicia; Arn, Kornelius; Matthes, Thomas; Hogan, Cassandra; Keller, Peter; Dudkiewicz, Ewa; Stüssi, Georg; Fernandez, Paula
2017-07-29
The EuroFlow Consortium developed a fully standardized flow cytometric approach from instrument settings, through antibody panel, reagents and sample preparation protocols, to data acquisition and analysis. The Swiss Cytometry Society (SCS) promoted a study to evaluate the feasibility of using such standardized measurements of 8-color data across two different flow cytometry platforms - Becton Dickinson (BD) FACSCanto II and Beckman Coulter (BC) Navios, aiming at increasing reproducibility and inter-laboratory comparability of immunophenotypic data in clinical laboratories in Switzerland. The study was performed in two phases, i.e. a learning phase (round 1) and an analytical phase (rounds 2 and 3) consisting of a total of three rounds. Overall, 10 laboratories using BD FACSCanto II (n=6) or BC Navios (n=4) flow cytometers participated. Each laboratory measured peripheral blood samples from healthy donors stained with a uniform antibody panel of reagents - EuroFlow Lymphoid Screening Tube (LST) - applying the EuroFlow standardized protocols for instrument setup and sample preparation (www.EuroFlow.org). All data files were analyzed centrally and median fluorescence intensity (MedFI) values for individual markers on defined lymphocyte subsets were recorded; variability from reference MedFI values was assessed using performance scores. Data troubleshooting and discussion of the results with the participants followed after each round at SCS meetings. The results of the learning phase demonstrated that standardized instrument setup and data acquisition are feasible in routine clinical laboratories without previous experience with EuroFlow. During the analytical phase, highly comparable data were obtained at the different laboratories using either BD FACSCanto II or BC Navios. The coefficient of variation of MedFI for 7 of 11 markers performed repeatedly below 30%. In the last study round, 89% of participants scored over 90% MedFI values within the acceptance criteria
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Erisgin, Z; Tekelioglu, Y
For epileptics, pregnancy contains the balance between no seizure period and antiepileptic use having the least teratogenicity risk. The purpose is to analyse with flow cytometry the apoptotic effects on postnatal brain tissue caused by prenatal use of second generation antiepileptics oxcarbazepine (OXC) and gabapentin (GBP) having different effect mechanisms. 30 (n = 5 each group) Wistar albino male rats (45-days-old) are used. First 3 groups are exposed to OXC (100 mg/kg/day), GBP (50 mg/kg/day), and saline, respectively on the 1st-5th prenatal days (preimplantation-implantation period) while the second 3 groups are exposed to the same substances on the 6th-15th prenatal days (organogenesis), respectively. After sacrifice, brain tissue samples were made into suspension with mechanic and enzymatic digestion and examined with flow cytometry. While apoptosis rate appeared high in rats exposed to OXC on the 1st-5th (p effect in three treatment groups, while difference was not significant for PSS and GBP groups (p = 0.847 and p = 0.934), apoptosis rate was significantly high for OXC on the 6th-15th days compared to the 1st-5th days (p < 0.001). It is observed that the use of OXC causes neurotoxicity during preimplantation, implantation and, especially, organogenesis period (neurogenesis) whereas GBP does not (Fig. 3, Ref. 32).
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[Computer-aided Prognosis for Breast Cancer Based on Hematoxylin & Eosin Histopathology Image].
Chen, Jiamei; Qu, Aiping; Liu, Wenlou; Wang, Linwei; Yuan, Jingping; Liu, Juan; Li, Yan
2016-06-01
Quantitatively analyzing hematoxylin &eosin(H&E)histopathology images is an emerging field attracting increasing attentions in recent years.This paper reviews the application of computer-aided image analysis in breast cancer prognosis.The traditional prognosis based on H&E histopathology image for breast cancer is firstly sketched,followed by a detailed description of the workflow of computer-aided prognosis including image acquisition,image preprocessing,regions of interest detection and object segmentation,feature extraction,and computer-aided prognosis.In the end,major technical challenges and future directions in this field are summarized.
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Claire L Anderson; Thomas L Kubisiak; C Dana Nelson; Jason A Smith; John M Davis
2010-01-01
The genome size of the pine fusiform rust pathogen Cronartium quercuum f.sp. fusiforme (Cqf) was determined by flow cytometric analysis of propidium iodide-stained, intact haploid pycniospores with haploid spores of two genetically well characterized fungal species, Sclerotinia sclerotiorum and Puccinia graminis f.sp. tritici, as size standards. The Cqf haploid genome...
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Santhanalakshmi, J.; Venkatesan, P.
2011-02-01
Mono metallic (Au, Ag, Pd) and bimetallic (Au-Ag, Ag-Pd, Au-Pd) with 1:1 mol stoichiometry, nanoparticles are synthesized using one-pot, temperature controlled chemical method using cetyltrimethylammonium bromide (CTAB) as the capping agent. The particle sizes (Au = 5.6, Ag = 5.0, Pd = 6.0, Au-Ag = 9.2, Ag-Pd = 9.6, Au-Pd = 9.4 nm) are characterized by UV-Vis, HRTEM, and XRD measurements, respectively. CTAB bindings onto mono and bimetallic nanoparticles are analyzed by FTIR spectra. The catalytic activities of mono and bimetallic nanoparticles are tested on the reaction between NADH oxidation and Eosin-Y reduction. The effects of base, pH, ionic strength, nature of mono and bimetallic catalysts are studied and the reaction conditions are optimized. Bimetallic nanoparticles exhibited better catalysis than the mono metallic nanoparticles, which may be due to the electronic effects of the core to shell metal atoms.
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DNA flow cytometric analysis in variable types of hydropic placentas
Directory of Open Access Journals (Sweden)
Fatemeh Atabaki pasdar
2015-05-01
Full Text Available Background: Differential diagnosis between complete hydatidiform mole, partial hydatidiform mole and hydropic abortion, known as hydropic placentas is still a challenge for pathologists but it is very important for patient management. Objective: We analyzed the nuclear DNA content of various types of hydropic placentas by flowcytometry. Materials and Methods: DNA ploidy analysis was performed in 20 non-molar (hydropic and non-hydropic spontaneous abortions and 20 molar (complete and partial moles, formalin-fixed, paraffin-embedded tissue samples by flow cytometry. The criteria for selection were based on the histopathologic diagnosis. Results: Of 10 cases histologically diagnosed as complete hydatiform mole, 9 cases yielded diploid histograms, and 1 case was tetraploid. Of 10 partial hydatidiform moles, 8 were triploid and 2 were diploid. All of 20 cases diagnosed as spontaneous abortions (hydropic and non-hydropic yielded diploid histograms. Conclusion: These findings signify the importance of the combined use of conventional histology and ploidy analysis in the differential diagnosis of complete hydatidiform mole, partial hydatidiform mole and hydropic abortion.
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International Nuclear Information System (INIS)
Ostakhov, S.S.; Kolosnitsyn, V.S.; Krasnogorskaya, N.N.; Kazakov, V.P.
2000-01-01
Quenching of terbium nitrate luminescence by anionic dye, eosin, in the presence of lithium perchlorate in sulfolane solutions was studied. Temperature dependence of terbium nitrate luminescence in sulfolane solutions in the presence of perchlorate anions were considered. The values of energy required for water molecular substitution in Tb 3+ ion coordination sphere for solvent molecule in electrolyte solution were ascertained [ru
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Overmodulation Control in the Optimization of a H-PDLC Device with Ethyl Eosin as Dye
Directory of Open Access Journals (Sweden)
Manuel Ortuño
2013-01-01
Full Text Available The response of a H-PDLC device is improved by means of a two-step method. First, component optimization—initiator system, crosslinker, and cosolvent—enables the diffraction efficiency of the hologram to be maximized. Second, the use of N-methyl-2-pyrrolidone in combination with N-vinyl-2-pyrrolidone prevents the overmodulation in photopolymers containing ethyl eosin.
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Directory of Open Access Journals (Sweden)
Reginaldo Cipullo
2011-08-01
Full Text Available FUNDAMENTO: O significado clínico de vasculites, lesões isquêmicas, efeito Quilty e da presença de eosinófilos em biópsias endomiocárdicas de receptores de transplante cardíaco com rejeições leves não foi ainda estabelecido. OBJETIVO: Verificar se esses achados histológicos encontrados nas biópsias endomiocárdicas (eosinófilos, vasculites, efeito Quilty e lesões isquêmicas são capazes de predizer rejeição aguda do enxerto. MÉTODOS: Foram reavaliadas 1.012 biópsias endomiocárdicas consecutivas; dessas, 939 foram classificadas como OR ou 1R pela Nomenclatura da Sociedade Internacional de Transplante de Coração e Pulmão de 2005, e divididas em dois grupos: (1 Biópsias preditoras: aquelas que precederam rejeição aguda; e (2 Biópsias não preditoras: aquelas que não precederam rejeição aguda. Comparamos a ocorrência dos seguintes achados histológicos: vasculites, lesões isquêmicas, efeito Quilty e eosinófilos por análise uni e multivariada entre os grupos. RESULTADOS: Após análise estatística verificou-se a presença de vasculite intensa e de eosinófilos como maiores preditores para rejeição aguda futura, apresentando respectivamente as seguintes razões de chance: 10,60 (IC95%: 3,62 - 31,06. p < 0,001 e 6,26 (IC95%: 3,16 - 12,43. p < 0,001. CONCLUSÃO: Vasculites intensas e eosinófilos em biópsias do miocárdio são os principais fatores preditores de rejeição aguda pós-transplante cardíaco.
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Sampling polyhexamethylene guanidine aerosols using eosin Y-coated glass beads
International Nuclear Information System (INIS)
Choi, Sang Hyun; Park, Seon Kyung; Kang, Hyun Joong; Kwon, Jung Hwan; Lee, Kyu Hong
2015-01-01
Fatalities caused by the use of polyhexamethylene guanidine (PHMG), a general-purpose chemical germicide used as a humidifier disinfectant in Korea, have raised concerns about exposure to biocide aerosols in indoor environments. A sampler capable of accumulating PHMG from aqueous aerosols was developed as an alternative to low-volume air samplers. This sampler was prepared by placing glass beads coated with 2-(2,4,5,7-tetrabromo-6-oxido-3-oxo-3H-xanthen-9-yl)benzoate (Eosin Y) in a custom-made plastic holder. Passive sampling rates, measured in a bench-top exposure chamber at two different aqueous PHMG aerosol generation rates, were found to be independent of the experimental conditions. This suggests that the capacity of the sampler to accumulate the PHMG aerosol was sufficient for the sampling duration tested. However, the passive sampling rate was 7.6 × 10"−"6 m"3/h for the sampler area of 22 cm"2. This rate is lower than the typical human breathing rate and inadequate for quantitative instrumental analyses at low concentrations in indoor air. A 30-fold enhancement of the sampling rate was achieved by forced convection using a commercial battery-operated fan at ≥2000 rpm. With this accelerated sampling rate, the sampler could be used to monitor time-integrated concentrations of PHMG aerosols in the air
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Sampling polyhexamethylene guanidine aerosols using eosin Y-coated glass beads
Energy Technology Data Exchange (ETDEWEB)
Choi, Sang Hyun; Park, Seon Kyung; Kang, Hyun Joong; Kwon, Jung Hwan [Korea University, Seoul (Korea, Republic of); Lee, Kyu Hong [Korea Institute of Toxicology, Jeongeup (Korea, Republic of)
2015-07-15
Fatalities caused by the use of polyhexamethylene guanidine (PHMG), a general-purpose chemical germicide used as a humidifier disinfectant in Korea, have raised concerns about exposure to biocide aerosols in indoor environments. A sampler capable of accumulating PHMG from aqueous aerosols was developed as an alternative to low-volume air samplers. This sampler was prepared by placing glass beads coated with 2-(2,4,5,7-tetrabromo-6-oxido-3-oxo-3H-xanthen-9-yl)benzoate (Eosin Y) in a custom-made plastic holder. Passive sampling rates, measured in a bench-top exposure chamber at two different aqueous PHMG aerosol generation rates, were found to be independent of the experimental conditions. This suggests that the capacity of the sampler to accumulate the PHMG aerosol was sufficient for the sampling duration tested. However, the passive sampling rate was 7.6 × 10{sup −6} m{sup 3}/h for the sampler area of 22 cm{sup 2}. This rate is lower than the typical human breathing rate and inadequate for quantitative instrumental analyses at low concentrations in indoor air. A 30-fold enhancement of the sampling rate was achieved by forced convection using a commercial battery-operated fan at ≥2000 rpm. With this accelerated sampling rate, the sampler could be used to monitor time-integrated concentrations of PHMG aerosols in the air.
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Energy Technology Data Exchange (ETDEWEB)
Imamura, Nobutaka; Kimura, Akiro [Hiroshima Univ. (Japan). Research Inst. for Radiation Biology and Medicine
1998-12-01
In 51 patients (atomic bomb survivors 50, unexposed persons 1) who have had neutropenia for two years or more under indistinct cause, cell surface antigen was analyzed by flow cytometry. Twenty-nine cases of survivors were diagnosed as NK cell leukemia or NK cell cytosis because analysis data showed CD3(-), CD56(+) and CD57(+/-). Six cases were diagnosed as NK like T cell hypercytosis because analysis data showed CD3(+), CD56(+/-) and CD57(+). As for 15 cases, CD56(+) cell number was in range of 15.96{+-}5.35 of a normal person, and no relation with NK cell was recognized. But, CD4/CD8 ratio was higher than 2.1, and gain of T helper cell was recognized. One unexposed persons was diagnosed as chronic NK cell leukemia because analysis data showed CD3(-), CD56(+) and CD57(+). Anti-neutrophil antibody wasn't recognized. Cytotoxic activity for K562 and Raji cell line showed high value compared with that of a normal person. Epstein Barr virus wasn't detected. (K.H.)
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Vlasova, I. M.; Saletsky, A. M.
2008-11-01
The comparative analysis of binding of three molecular fluorescent probes (fluorescein, eosin, and erythrosin), belonging to one homologous family, to human serum albumin (HSA) is made by Raman spectroscopy method. The binding of all three probes to binding Center I of HSA is registered. The character of binding of initial probe of the given homologous family - fluorescein - to protein differs from character of binding of its halogen-derivatives (eosin and erythrosin) to protein. The differences in binding of these three probes to HSA are determined by value of electronegativity of atoms of lateral radicals in structural formulas of probes and, therefore, by value of pK of their ionized groups.
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Energy Technology Data Exchange (ETDEWEB)
Vera, F. [Instituto de Quimica, Universidad Catolica de Valparaiso, Valparaiso (Chile); Schrebler, R. [Instituto de Quimica, Universidad Catolica de Valparaiso, Valparaiso (Chile); Munoz, E. [Instituto de Quimica, Universidad Catolica de Valparaiso, Valparaiso (Chile); Suarez, C. [Instituto de Quimica, Universidad Catolica de Valparaiso, Valparaiso (Chile); Cury, P. [Instituto de Quimica, Universidad Catolica de Valparaiso, Valparaiso (Chile); Gomez, H. [Instituto de Quimica, Universidad Catolica de Valparaiso, Valparaiso (Chile); Cordova, R. [Instituto de Quimica, Universidad Catolica de Valparaiso, Valparaiso (Chile); Marotti, R.E. [Instituto de Fisica, Facultad de Ingenieria, Universidad de la Republica, Herrera y Reissig 565, C.C. 30, 11000 Montevideo (Uruguay); Dalchiele, E.A. [Instituto de Fisica, Facultad de Ingenieria, Universidad de la Republica, Herrera y Reissig 565, C.C. 30, 11000 Montevideo (Uruguay)]. E-mail: dalchiel@fing.edu.uy
2005-11-01
Nickel oxide (NiO) thin films were prepared onto ITO/glass substrates by spin-coating, dipping and electrochemically. Studies of the morphological and structural properties of the films were done by atomic force microscopy (AFM). Photoelectrochemical and optical experiments were carried out in order to characterize the semiconductor properties of the nanostructured NiO thin films. The experiments were also done for Eosin B- and Erythrosin J-sensitized nanostructured NiO films, with the aim to visualize their potential application as photocatodes in tandem dye-sensitized solar cells (TDSSC). The NiO grown by dipping was the one presenting the best morphological properties. The photoelectrochemical results for all the bare NiO, NiO-Eosin B and NiO-Erythrosin J/electrolyte (I{sub 2}/I{sup -}) systems showed a p-type behavior. An enhancement in the photocurrent has been observed for the systems sensitized with the dyes. For the NiO/Erythrosin J system the enhancement of the current under illumination in comparison to the dark current was about 200%.
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International Nuclear Information System (INIS)
Vera, F.; Schrebler, R.; Munoz, E.; Suarez, C.; Cury, P.; Gomez, H.; Cordova, R.; Marotti, R.E.; Dalchiele, E.A.
2005-01-01
Nickel oxide (NiO) thin films were prepared onto ITO/glass substrates by spin-coating, dipping and electrochemically. Studies of the morphological and structural properties of the films were done by atomic force microscopy (AFM). Photoelectrochemical and optical experiments were carried out in order to characterize the semiconductor properties of the nanostructured NiO thin films. The experiments were also done for Eosin B- and Erythrosin J-sensitized nanostructured NiO films, with the aim to visualize their potential application as photocatodes in tandem dye-sensitized solar cells (TDSSC). The NiO grown by dipping was the one presenting the best morphological properties. The photoelectrochemical results for all the bare NiO, NiO-Eosin B and NiO-Erythrosin J/electrolyte (I 2 /I - ) systems showed a p-type behavior. An enhancement in the photocurrent has been observed for the systems sensitized with the dyes. For the NiO/Erythrosin J system the enhancement of the current under illumination in comparison to the dark current was about 200%
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Flow Cytometric Applicability of Fluorescent Vitality Probes on Phytoplankton
Peperzak, L.; Brussaard, C.P.D.
2011-01-01
The applicability of six fluorescent probes (four esterase probes: acetoxymethyl ester of Calcein [Calcein-AM], 5-chloromethylfluorescein diacetate [CMFDA], fluorescein diacetate [FDA], and 2',7'-dichlorofluorescein diacetate [H(2)DCFDA]; and two membrane probes: bis-(1,3-dibutylbarbituric acid)
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Improved performance of a dye-sensitized solar cell using a TiO{sub 2}/ZnO/Eosin Y electrode
Energy Technology Data Exchange (ETDEWEB)
Kim, Seok-Soon; Yum, Jun-Ho; Sung, Yung-Eun [Department of Materials Science and Engineering, Kwangju Institute of Science and Technology(K-JIST), Kwangju 500-712 (Korea, Republic of)
2003-09-30
TiO{sub 2}/ZnO/Eosin Y structure films were prepared by a one-step cathodic electrodeposition method and used as a photoanode in a dye-sensitized solar cell (DSSC). Using this TiO{sub 2}/ZnO/Eosin Y electrode in DSSC, the degradation of the cell with time was reduced and I{sub SC}, V{sub OC} and fill factor values were increased. The use of a thin ZnO layer, permitted the formation of an energy barrier at the electrode/electrolyte interface, thus reducing recombination rate and improving cell performance. In addition, the adsorbed dye molecules prepared by one-step cathodic electrodeposition with ZnO were very stable compared with that prepared by conventional immersing method, as evidenced by UV/vis absorption spectroscopy measurements.
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Improved performance of a dye-sensitized solar cell using a TiO{sub 2}/ZnO/Eosin Y electrode
Energy Technology Data Exchange (ETDEWEB)
Seoksoon Kim; Junho Yum; Yungeun Sung [Kwangju Inst. of Science and Technology (K-JIST), Dept. of Materials Science and Engineering, Kwangju (Korea)
2003-09-30
TiO{sub 2}/ZnO/Eosin Y structure films were prepared by a one-step cathodic electrodeposition method and used as a photoanode in a dye-sensitized solar cell (DSSC). Using this TiO{sub 2}/ZnO/Eosin Y electrode in DSSC, the degradation of the cell with time was reduced and I{sub sc}, V{sub oc} and fill factor values were increased. The use of a thin ZnO layer, permitted the formation of an energy barrier at the electrode/electrolyte interface, thus reducing recombination rate and improving cell performance. In addition, the adsorbed dye molecules prepared by one-step cathodic electrodeposition with ZnO were very stable compared with that prepared by conventional immersing method, as evidenced by UV /vis absorption spectroscopy measurements. (Author)
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Synthesis of 2,4,6-Trisubstituted Pyridines by Oxidative Eosin Y Photoredox Catalysis.
Rohokale, Rajendra S; Koenig, Burkhard; Dhavale, Dilip D
2016-08-19
Eosin Y, an organic dye, was activated as a photoredox catalyst in the presence of molecular oxygen using visible light and, when it was used in the reaction of aryl ketones and benzyl amines, afforded good yields (52-87%) of 2,4,6-triarylpyridines (21 examples) at ambient temperature. The aryl groups at the 2- and 6-positions are derived from ketones, while benzyl amine plays the dual role of providing an aryl functionality at the 4-position of pyridine as well as being a nitrogen donor.
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Yin, Yao; Weng, Wei-Zhi; Sun, Jian-Guo; Zhang, Bo
2018-03-28
A visible-light-promoted phosphinylation of allylic alcohols with concomitant 1,2-aryl migration is described. This transformation proceeds smoothly under metal-free and mild conditions by using an inexpensive organic dye, eosin Y, as the photocatalyst, affording various β-aryl-γ-ketophosphine oxides in moderate to good yields. Mechanistic studies suggested that the 1,2-aryl migration proceeded through a radical (neophyl) rearrangement.
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A flow cytometric method for characterization of circulating cell-derived microparticles in plasma
DEFF Research Database (Denmark)
Nielsen, Morten Hjuler; Beck-Nielsen, Henning; Andersen, Morten Nørgaard
2014-01-01
BACKGROUND AND AIM: Previous studies on circulating microparticles (MPs) indicate that the majority of MPs are of a size below the detection limit of most standard flow cytometers. The objective of the present study was to establish a method to analyze MP subpopulations above the threshold...
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... people who don't have FA. Cytometric Flow Analysis Cytometric flow analysis, or CFA, is done in a lab. This ... to know about resources that can give you emotional support and helpful information about FA and its ...
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DEFF Research Database (Denmark)
Knaus, Alexej; Pantel, Jean Tori; Pendziwiat, Manuela
2018-01-01
, the increasing number of individuals with a GPIBD shows that hyperphosphatasia is a variable feature that is not ideal for a clinical classification. METHODS: We studied the discriminatory power of multiple GPI-linked substrates that were assessed by flow cytometry in blood cells and fibroblasts of 39 and 14...... those with PIGA mutations. Although the impairment of GPI-linked substrates is supposed to play the key role in the pathophysiology of GPIBDs, we could not observe gene-specific profiles for flow cytometric markers or a correlation between their cell surface levels and the severity of the phenotype...
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DEFF Research Database (Denmark)
Dalgaard, T. S.; Norup, L. R.; Pedersen, A.R.
2010-01-01
. Despite a delayed NDV-specific antibody response to vaccination, L133 appeared to be better protected than L130 in the subsequent infection challenge as determined by the presence of viral genomes. Peripheral blood was analyzed by flow cytometry and responses in vaccinated/challenged birds were studied...... by 5-color immunophenotyping as well as by measuring the proliferative capacity of NDV-specific T cells after recall stimulation. Immunophenotyping identified L133 as having a significantly lower CD4/CD8 ratio and a lower frequency of γδ T cells than L130 in the peripheral T cell compartment...
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Directory of Open Access Journals (Sweden)
A. M. Popov
2015-01-01
Full Text Available 191 consecutive unselected children with acute lymphoblastic leukemia aged from 1 to 16 years were enrolled in the study. Bone marrow samples were obtained at the time of initial diagnostics as well as at days 15 (n = 188, 36 (n = 191, and 85 (n = 187 of remission induction. Minimal residual disease (MRD was assessed by 6–10-color flow cytometry. Flow cytometry data at day 15 allowed distinguishing three patients groups with significantly different outcome (p ˂ 0.0001: 35.64 % patients with MRD < 0.1 % represented 5-year event-free survival (EFS of 100 %; 48.40 % cases with 0.1 % ≤ MRD< 10 % had EFS 84.6 ± 4.2 %; 15.96 % patients with very high MRD (≥ 10 % belonged to group with poor outcome (EFS 56.7 ± 9.0 %. At the end of remission induction (day 36 36 children (18.85 % with MRD higher than 0.1 % had significantly worse outcome compared to remaining ones (EFS 49.4 ± 9.0 and 93.5 ± 2.1 % respectively; p ˂ 0.0001. From a clinical standpoint it is relevant to evaluate both low-risk and high-risk criteria. Multivariate analysis showed that day 15 MRD data is better for low-risk patients definition while end-induction MRD is the strongest unfavorable prognostic factor.
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International Nuclear Information System (INIS)
Santhanalakshmi, J.; Venkatesan, P.
2011-01-01
Mono metallic (Au, Ag, Pd) and bimetallic (Au–Ag, Ag–Pd, Au–Pd) with 1:1 mol stoichiometry, nanoparticles are synthesized using one-pot, temperature controlled chemical method using cetyltrimethylammonium bromide (CTAB) as the capping agent. The particle sizes (Au = 5.6, Ag = 5.0, Pd = 6.0, Au–Ag = 9.2, Ag–Pd = 9.6, Au–Pd = 9.4 nm) are characterized by UV–Vis, HRTEM, and XRD measurements, respectively. CTAB bindings onto mono and bimetallic nanoparticles are analyzed by FTIR spectra. The catalytic activities of mono and bimetallic nanoparticles are tested on the reaction between NADH oxidation and Eosin-Y reduction. The effects of base, pH, ionic strength, nature of mono and bimetallic catalysts are studied and the reaction conditions are optimized. Bimetallic nanoparticles exhibited better catalysis than the mono metallic nanoparticles, which may be due to the electronic effects of the core to shell metal atoms.Graphical Abstract
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Coppo, Lucia; Montano, Sergio J; Padilla, Alicia C; Holmgren, Arne
2016-04-15
Glutaredoxins catalyze glutathione-dependent disulfide oxidoreductions, particularly reduction of glutathione (GSH)-protein mixed disulfides. Mammalian glutaredoxins are present in the cytosol/nucleus as Grx1 or in mitochondria as Grx2a. Here we describe di-eosin-glutathione disulfide (Di-E-GSSG) as a new tool to study glutaredoxin (Grx) activity. Di-E-GSSG has almost no fluorescence in its disulfide form due to self-quenching, whereas the reduced form (E-GSH) has a large fluorescence emission at 545 nm after excitation at 520 nm. Di-E-GSSG was a very poor substrate for glutathione reductase, but we discovered that the molecule was an excellent substrate for glutaredoxin in a coupled assay system with GSH, nicotinamide adenine dinucleotide phosphate (NADPH), and glutathione reductase or with lipoamide, NADH, and lipoamide dehydrogenase. In addition, Di-E-GSSG was used to glutathionylate the free SH group of bovine serum albumin (BSA), yielding eosin-glutathionylated BSA (E-GS-BSA) readily observed in ultraviolet (UV) light. E-GS-BSA also displayed a quenched fluorescence, and its Grx-catalyzed reduction could be followed by the formation of E-GSH by fluorescence emission using microtiter plates. This way of measuring Grx activity provided an ultrasensitive method that detected Grx1 and Grx2 at picomolar levels. Human Grx1 was readily quantified in 40 μl of plasma and determined to be 680 ± 208 pM in healthy controls. Copyright © 2016 Elsevier Inc. All rights reserved.
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Comparison of Five Nuclear Isolation Buffers for Flow Cytometric ...
African Journals Online (AJOL)
Jocky
2012-02-23
Feb 23, 2012 ... species. A systematic comparison of nuclear lysis buffers has been ... All experiments were carried out with 3 replicates (n = 3) per treatment. ... Na2EDTA, 0.5 mM spermine.4HCl, 80 mM KCl, 20 mM NaCl, 0.1%. (v/v) Triton ...
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Ultrastructural and flow cytometric analyses of lipid accumulation in microalgae
Energy Technology Data Exchange (ETDEWEB)
Solomon, J.A.; Hand, R.E. Jr.; Mann, R.C.
1986-12-01
Lipid accumulation in three species of microalgae was investigated with flow cytometry (FCM) and transmission electron microscopy (TEM). Previous studies using batch cultures of a algae have led to the assumption that lipid accumulation in microalgae is a gradual process requiring at least several days for completion. However, FCM reveals, through changes in the chlorophyll:lipid ratio, that the time span required for individual cells to change metabolic state is short. Simultaneous FCM measurements of chlorophyll and nile red (neutral lipid) fluorescence in individual cells of nitrogen-deficient Isochrysis populations revealed a bimodal population distribution as one stage in the lipid accumulation process. The fact that two discrete populations exist, with few cells in an intermediate stage, suggests rapid response to a liqid trigger. Interpretations of light and electron microscopic observations are consistent with this hypothesis. The time required for an entire population to achieve maximum lipid content is considerably longer than that required for a single cell, due to the variation in response time among cells. In this study high lipid cultures were sometimes obtained by using FCM to separate high lipid cells from the remainder of the population. FCM holds much promise for strain enhancement but considerable developmental work, directed at providing more consistent results, remains to be done. 8 refs., 35 figs.
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Directory of Open Access Journals (Sweden)
Jing Yan
2013-12-01
Full Text Available Here we report the application of monodisperse double emulsion droplets, produced in a single step within partially hydrophilic/partially hydrophobic microfluidic devices, as defined containers for quantitative flow cytometric analysis. Samples with varying fluorophore concentrations were generated, and a clear correlation between dye concentration and fluorescence signals was observed.
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McLenachan, Samuel; Sarsero, Joseph P; Ioannou, Panos A
2007-06-01
Using the lipofection reagent LipofectAMINE 2000 we have examined the delivery of plasmid DNA (5-200 kb) to mouse embryonic stem (mES) cells by flow cytometry. To follow the physical uptake of lipoplexes we labeled DNA molecules with the fluorescent dye TOTO-1. In parallel, expression of an EGFP reporter cassette in constructs of different sizes was used as a measure of nuclear delivery. The cellular uptake of DNA lipoplexes is dependent on the uptake competence of mES cells, but it is largely independent of DNA size. In contrast, nuclear delivery was reduced with increasing plasmid size. In addition, linear DNA is transfected with lower efficiency than circular DNA. Inefficient cytoplasmic trafficking appears to be the main limitation in the nonviral delivery of large DNA constructs to the nucleus of mES cells. Overcoming this limitation should greatly facilitate functional studies with large genomic fragments in embryonic stem cells.
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A novel hematoxylin and eosin stain assay for detection of the parasitic dinoflagellate Amoebophrya.
Li, Caiwen; Chen, Tiantian
2017-02-01
The parasitic dinoflagellate Amoebophrya infects broad range of marine organisms. Particularly, Amoebophrya infections in planktonic dinoflagellates can prevent or delay the formation of algal blooms, and recycle undergrazed planktonic dinoflagellates back to the microbial loop by disrupting host cells. Its ecological significance was gradually recognized along with the discovery of its enormous molecular diversity in oceanic and coastal ecosystems. Thus, we developed a reliable, easily accessible and less time-consuming assay, to detect and assess Amoebophrya infections in planktonic dinoflagellates. The modified hematoxylin and eosin staining assay provided reliable diagnosis of Amoebophrya infection by identifying the characteristic "beehive" of the multinucleate trophonts. After staining, the typical multinucleate "beehive" is evidently distinguishable from the compact nuclei of uninfected host cells. The modified hematoxylin and eosin (H & E) staining assay is easy to use, that can be routinely performed within 3h (up to 20 samples/batch) using general laboratory equipment, supplies and chemical reagents. The produced slides with agar-embedded dinoflagellate cells can be stored for several months or even years in a dry place without noticeable loss in quality of staining. With suitable calculation, the modified H & E assay can be applied to assess the prevalence of Amoebophrya infection in planktonic dinoflagellates. This efficient and powerful assay will facilitate the investigation on the ecological roles of Amoebophryidae in coastal and oceanic ecosystem. Copyright © 2016 Elsevier B.V. All rights reserved.
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Energy Technology Data Exchange (ETDEWEB)
Kashiwado, Kouzou
1988-12-01
Radiosensitivity, Thermosensitivity and their combined sensitivity on cell death were studied using mouse L5178Y, mouse FM3A and human Burkitt lymphoma. Eosin-staining method was tested for counting the heat-induced dead cells. The results obtained in the present study were as follows: (1) In irradiation with /sup 60/Co gamma-rays D/sub 0/ values of 1.3 Gy, 1.4 Gy and 1.7 Gy were obtained for Burkitt lymphoma, L5178Y and FM3A respectively. (2) By heating at 43degC, T/sub 0/ values of 4.1 min, 12.5 min and 32 min were obtained for L5178Y, FM3A and Burkitt lymphoma respectively. (3) Thermotolerance decay depended on cell doubling time, that is, the shorter the doubling time, the faster the decay. Furthermore, the cell line with the higher thermosensitivity showed a faster decay of thermotolerance for the three cell lines used in the present study. (4) The radiosensitizing effects were nearly the same in all three cell lines. (5) Using eosin-staining method, the cell survival curves after hyperthermia were ascertained and compared with those obtained by colony forming method. In L5178Y the survival curves obtained by the two methods were nearly the same. In FM3A only some correlation was found betweewn the cell survival curves obtained by the two methods but in Burkitt lymphoma no correlation was found. (author) 52 refs.
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International Nuclear Information System (INIS)
Kashiwado, Kouzou
1988-01-01
Radiosensitivity, Thermosensitivity and their combined sensitivity on cell death were studied using mouse L5178Y, mouse FM3A and human Burkitt lymphoma. Eosin-staining method was tested for counting the heat-induced dead cells. The results obtained in the present study were as follows: 1) In irradiation with 60 Co gamma-rays D 0 values of 1.3 Gy, 1.4 Gy and 1.7 Gy were obtained for Burkitt lymphoma, L5178Y and FM3A respectively. 2) By heating at 43degC, T 0 values of 4.1 min, 12.5 min and 32 min were obtained for L5178Y, FM3A and Burkitt lymphoma respectively. 3) Thermotolerance decay depended on cell doubling time, that is, the shorter the doubling time, the faster the decay. Furthermore, the cell line with the higher thermosensitivity showed a faster decay of thermotolerance for the three cell lines used in the present study. 4) The radiosensitizing effects were nearly the same in all three cell lines. 5) Using eosin-staining method, the cell survival curves after hyperthermia were ascertained and compared with those obtained by colony forming method. In L5178Y the survival curves obtained by the two methods were nearly the same. In FM3A only some correlation was found betweewn the cell survival curves obtained by the two methods but in Burkitt lymphoma no correlation was found. (author) 52 refs
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5 X 5 rod bundle flow field measurements downstream a PWR spacer grid
Energy Technology Data Exchange (ETDEWEB)
Castro, Higor F.P.; Silva, Vitor V A.; Santos, André A.C.; Veloso, Maria A.F., E-mail: higorfabiano@gmail.com, E-mail: mdora@nuclear.ufmg.br, E-mail: vitors@cdtn.br, E-mail: aacs@cdtn.br [Universidade Federal de Minas Gerais (UFMG), Belo Horizonte, MG (Brazil); Centro de Desenvolvimento da Tecnologia Nuclear (CDTN/CNEN-MG), Belo Horizonte, MG (Brazil)
2017-07-01
The spacer grids are structures present in nuclear fuel assembly of Pressurized Water Reactors (PWR). They play an important structural role and also assist in heat removal through the assembly by promoting increased turbulence of the flow. Understanding the flow dynamics downstream the spacer grid is paramount for fuel element design and analysis. This paper presents water flow velocity profiles measurements downstream a spacer grid in a 5 x 5 rod bundle test rig with the objective of highlighting important fluid dynamic behavior near the grid and supplying data for CFD simulation validation. These velocity profiles were obtained at two different heights downstream the spacer grid using a LDV (Laser Doppler Velocimetry) through the top of test rig. The turbulence intensities and patterns of the swirl and cross flow were evaluated. The tests were conducted for Reynolds numbers ranging from 1.8 x 10{sup 4} to 5.4 x 10{sup 4}. This experimental research was carried out in thermo-hydraulics laboratory of Nuclear Technology Development Center – CDTN. Results show great repeatability and low uncertainties (< 1.24 %). Details of the flow field show how the mixture and turbulence induced by the spacer grid quickly decays downstream the spacer grid. It is shown that the developed methodology can supply high resolution low uncertainty results that can be used for validation of CFD simulations. (author)
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5 X 5 rod bundle flow field measurements downstream a PWR spacer grid
International Nuclear Information System (INIS)
Castro, Higor F.P.; Silva, Vitor V A.; Santos, André A.C.; Veloso, Maria A.F.
2017-01-01
The spacer grids are structures present in nuclear fuel assembly of Pressurized Water Reactors (PWR). They play an important structural role and also assist in heat removal through the assembly by promoting increased turbulence of the flow. Understanding the flow dynamics downstream the spacer grid is paramount for fuel element design and analysis. This paper presents water flow velocity profiles measurements downstream a spacer grid in a 5 x 5 rod bundle test rig with the objective of highlighting important fluid dynamic behavior near the grid and supplying data for CFD simulation validation. These velocity profiles were obtained at two different heights downstream the spacer grid using a LDV (Laser Doppler Velocimetry) through the top of test rig. The turbulence intensities and patterns of the swirl and cross flow were evaluated. The tests were conducted for Reynolds numbers ranging from 1.8 x 10"4 to 5.4 x 10"4. This experimental research was carried out in thermo-hydraulics laboratory of Nuclear Technology Development Center – CDTN. Results show great repeatability and low uncertainties (< 1.24 %). Details of the flow field show how the mixture and turbulence induced by the spacer grid quickly decays downstream the spacer grid. It is shown that the developed methodology can supply high resolution low uncertainty results that can be used for validation of CFD simulations. (author)
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Tang, Yufang; Chen, Hao; Weng, Chao; Tang, Xiaohui; Zhang, Miaoling; Yang, Qiongqiong; Hu, Tao; Cai, Changqun
A new simple and highly sensitive catalytic kinetic method for the determination of trace amount of FA in food sample has been established. The method was based on the catalytic effect of FA on the oxidation of eosin Y by potassium bromate in present of phosphoric acid. The reaction was monitored spectrophotometrically by measuring the decrease in absorbance of eosin Y at 518 nm. Under the optimized experimental conditions, the developed method allowed the determination of FA in the range of 0.03-0.6 μg mL-1 with a good precision, and the limit of detection was down to 0.00988 μg mL-1. The relative standard deviation of five replicate measurements for the determination of FA in concentration 0.12 μg mL-1 was 1.8%. The proposed method was successfully applied to the determination of FA in food directly and satisfactory results were obtained.
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Hirschsprung's disease diagnosis: Comparison of immunohistochemical, hematoxilin and eosin staining
Memarzadeh, Mehrdad; Talebi, Ardeshir; Edalaty, Masod; Hosseinpour, Mehrdad; Vahidi, Nasrin
2009-01-01
Background: The diagnosis of Hirschsprung's disease (HD) is based on the absence of ganglion cells. In hemotoxilin and eosin (H and E) as well as acetylcholine esterase staining there are limitations in the diagnosis of immature ganglion cells in neonates. Methods: In this prospective study, 54 biopsies taken from suspected HD patients (five mucosal specimens and 49 full thickness specimens) were studied. In the laboratory, after preparing sections of paraffin embedded tissues, H and E staining slides were compared with immunohistochemical (IHC) staining including: S100, NSE, CD117, CD56, Cathepsin D, Vimentin, BCL2, GFAP, Synaptophysin and chromogranin. Results: The study revealed 30 negative (absence of ganglion cells) cases (55.5%), 17 positive cases (31.04%) and seven suspected cases (12.9%) of ganglion cells on the H and E staining. On IHC staining with CD56 and Cathepsin D, all of the 17 positive cases detected through H and E, were confirmed for having ganglion cells and out of 30 cases reported negative on H and E staining, 28(93.3%) were reported negative and two (6.7%) positive by IHC staining. Of the seven suspected cases H and E staining), IHC staining detectedganglion cells only in five slides; two remained negative. Conclusions: IHC staining using CD56 and Cathepsin D improved the accuracy of diagnosis in HD when used in addition to H and E staining technique, especially for negative or suspicious slides. PMID:20671847
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International Nuclear Information System (INIS)
Hosono, Eiji; Fujihara, Shinobu; Kimura, Toshio
2004-01-01
Micro/nano-textured ZnO thick films were synthesized through deposition and pyrolysis of layered hydroxide zinc acetate (LHZA), Zn 5 (OH) 8 (CH 3 COO) 2 ·2H 2 O. LHZA films having a unique, rose-like morphology were initially deposited on conducting glass sheets in a chemical bath composed of methanol and zinc acetate dihydrate at 60 deg. C under neutral conditions. Pyrolysis of the LHZA films resulted in formation of ZnO without destroying the original morphology. Pyrolysis temperatures were found to greatly influence grain sizes and specific surface areas of the ZnO films. Photoelectrochemical performance of the films as ZnO/eosin Y electrodes was investigated in dye-sensitized solar cells using an I - /I 3 - redox electrolyte solution. The cell using the ZnO film pyrolyzed at 150 deg. C exhibited overall light to electricity conversion efficiencies of 2.0 and 3.3% under an AM-1.5 illumination at 100 and 10 mW cm -2 , respectively. While microscale pores in the electrodes facilitated mass transfer of fluid electrolytes in the depth direction, nanoscale pores contributed to an increase in the amount of adsorbed dye. The maximum incident photon-to-current conversion efficiency (IPCE) of the electrode reached 84.9% at a wavelength of 530 nm
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Directory of Open Access Journals (Sweden)
Umbertina Conti Reed
1975-06-01
Full Text Available É relatado um caso de granuloma eosinófilo da coluna cervical em menino branco de 10 anos de idade que apresentava dor cervical na altura de C5-C6, hemiparesia direita que evoluiu ulteriormente para tetraparesia, mista, de tipo motonêuronio inferior para membros superiores e de tipo motonêuronio superior para membros inferiores. O exame do líquido cefalorraquano (LCR revelou alterações de tipo inflamatório. Radiografias da coluna cervical mostravam apenas aspecto irregular da apófise espinhosa de C6 com rarefação na base. Houve acentuada e rápida melhora do quadro neurológico e normalização do LCR na vigência de córticoterapia e, reagravamento da sintomatologia neurológica bem como das alterações no LCR após supressão da referida medicação. A biópsia do processo espinhoso da sexta vértebra cervical revelou tratar-se de granuloma eosinófilo. A evolução foi excelente após radioterapia local associada a corticosteróides. São discutidas as características do caso e daqueles referidos na literatura.
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Eosin Y Catalyzed Visible-light-promoted One –Pot Facile Synthesis of 1,3,4- Thiadiazole
Directory of Open Access Journals (Sweden)
Vishal Srivastava
2015-03-01
Full Text Available A novel one-pot visible light irradiated synthesis of 1,3,4-thiadiazole from aldehydes and thioacyl hydrazides have been reported in presence of eosin Y as an organophotoredox catalyst at room temperature under aerobic condition. This synthesis includes application of air and visible light as inexpensive, readily available, non-toxic and sustainable regents, which fulfils the basic principle of green chemistry.
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Flow-cytometric measurement of CD4-8- T cells bearing T-cell receptor αβ chains, 1
International Nuclear Information System (INIS)
Kusunoki, Yoichiro; Hirai, Yuko; Kyoizumi, Seishi; Akiyama, Mitoshi.
1992-09-01
In this study we detected rare, possibly abnormal, T cells bearing CD3 surface antigen and T-cell receptor (TCR) αβ chains but lacking both CD4 and CD8 antigens (viz., TCRαβ + CD4 - 8 - cells, as determined by flow cytometry). The TCRαβ + CD4 - 8 - T cells were detected at a mean frequency of 0.63 ± 0.35 % (mean ± standard deviation) in peripheral blood TCRαβ + cells of 119 normal persons. Two unusual cases besides the 119 normal persons showed extremely elevated frequencies of TCRαβ + CD4 - 8 - T cells, viz., approximately 5 % to 10 % and 14 % to 19 % in whole TCRαβ + cells. Both individuals were males who were otherwise physiologically quite normal with no history of severe illness, and these high frequencies were also observed in blood samples collected 2 or 8 years prior to the current measurements. The TCRαβ + CD4 - 8 - T cells of the two individuals were found to express mature T-cell markers such as CD2,3, and 5 antigens, as well as natural killer (NK) cell markers, viz., CD11b, 16, 56, and 57 antigens, when peripheral blood lymphocytes were subjected to three-color flow cytometry. Lectin-dependent or redirected antibody-dependent cell-mediated cytotoxicities were observed for both freshly sorted TCRαβ + CD4 - 8 - cells and in vitro established clones. Nevertheless, NK-like activity was not detected. Further, Southern blot analysis of TCRβ and γ genes revealed identical rearrangement patterns for all the TCRαβ + CD4 - 8 - clones established in vitro. These results suggest that the TCRαβ + CD4 - 8 - T cells from these two mean exhibit unique characteristics and proliferate clonally in vivo. (author)
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Meamar, Mehrdad; Zribi, Nassira; Cambi, Marta; Tamburrino, Lara; Marchiani, Sara; Filimberti, Erminio; Fino, Maria Grazia; Biggeri, Annibale; Menezo, Yves; Forti, Gianni; Baldi, Elisabetta; Muratori, Monica
2012-08-01
To analyze the effect of cryopreservation on sperm DNA fragmentation (SDF) in two cytometric sperm populations, PI(brighter) and PI(dimmer), and to test the effects of Opuntia ficus-indica (OFI) extracts, which contain antioxidants and flavanoids, and of resveratrol on cryopreservation of human semen. In vitro prospective study. Institutional study. Twenty-one normozoospermic men undergoing semen analysis for couple infertility. Cryopreservation using the routine method in the presence of OFI extracts or resveratrol. Measurement of SDF by TUNEL/PI flow cytometric method to evaluate sperm motility (by automated motion analysis, CASA system) and viability (by eosin/nigrosin staining) in the two populations of sperm PI(br) and PI(dim). Cryopreservation induced an increase of SDF only in the PI(br) sperm population. The increase was negatively dependent on the basal values of SDF in the same population. Addition of OFI extracts and resveratrol to the cryopreservation medium slightly but statistically significantly reduced SDF in the PI(br) population without affecting the deleterious effect of cryopreservation on sperm motion parameters or viability. The increase of SDF in the PI(br) population, which is unrelated to semen quality, suggests that caution must be taken in using cryopreserved semen, as morphologically normal and motile sperm may be damaged. The addition of substances with multifunctional properties such as OFI extracts to cryopreservation medium is only slightly effective in preventing the dramatic effects on SDF. Copyright © 2012 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.
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International Nuclear Information System (INIS)
Dragojevic-Simic, V.; Colic, M.; Gasic, S.
1994-01-01
The T cell composition of the thymus of X-ray irradiated (3.5 Gy) Wistar rat protected with WR-638 was analyzed by flow cytometry using monoclonal antibodies directed to the Thy 1.1, CD43, CD2, CD5, CD4, CD8 and class I and II MHC antigens. It was shown that this dose of X-rays caused cyclic changes in thymic cellularity manifested as: primary involution (until day 2), primary regeneration (from days 2 to 14), secondary involution (from days 14 to 21) and secondary regeneration (from days 21 to 30). WR-638 reduced the magnitude of thymocyte depletion in the primary involutive phase of the irradiated thymi. (author)
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Lahiani, Amal; Klaiman, Eldad; Grimm, Oliver
2018-01-01
Medical diagnosis and clinical decisions rely heavily on the histopathological evaluation of tissue samples, especially in oncology. Historically, classical histopathology has been the gold standard for tissue evaluation and assessment by pathologists. The most widely and commonly used dyes in histopathology are hematoxylin and eosin (H&E) as most malignancies diagnosis is largely based on this protocol. H&E staining has been used for more than a century to identify tissue characteristics and structures morphologies that are needed for tumor diagnosis. In many cases, as tissue is scarce in clinical studies, fluorescence imaging is necessary to allow staining of the same specimen with multiple biomarkers simultaneously. Since fluorescence imaging is a relatively new technology in the pathology landscape, histopathologists are not used to or trained in annotating or interpreting these images. To allow pathologists to annotate these images without the need for additional training, we designed an algorithm for the conversion of fluorescence images to brightfield H&E images. In this algorithm, we use fluorescent nuclei staining to reproduce the hematoxylin information and natural tissue autofluorescence to reproduce the eosin information avoiding the necessity to specifically stain the proteins or intracellular structures with an additional fluorescence stain. Our method is based on optimizing a transform function from fluorescence to H&E images using least mean square optimization. It results in high quality virtual H&E digital images that can easily and efficiently be analyzed by pathologists. We validated our results with pathologists by making them annotate tumor in real and virtual H&E whole slide images and we obtained promising results. Hence, we provide a solution that enables pathologists to assess tissue and annotate specific structures based on multiplexed fluorescence images.
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Directory of Open Access Journals (Sweden)
Amal Lahiani
2018-01-01
Full Text Available Context: Medical diagnosis and clinical decisions rely heavily on the histopathological evaluation of tissue samples, especially in oncology. Historically, classical histopathology has been the gold standard for tissue evaluation and assessment by pathologists. The most widely and commonly used dyes in histopathology are hematoxylin and eosin (H&E as most malignancies diagnosis is largely based on this protocol. H&E staining has been used for more than a century to identify tissue characteristics and structures morphologies that are needed for tumor diagnosis. In many cases, as tissue is scarce in clinical studies, fluorescence imaging is necessary to allow staining of the same specimen with multiple biomarkers simultaneously. Since fluorescence imaging is a relatively new technology in the pathology landscape, histopathologists are not used to or trained in annotating or interpreting these images. Aims, Settings and Design: To allow pathologists to annotate these images without the need for additional training, we designed an algorithm for the conversion of fluorescence images to brightfield H&E images. Subjects and Methods: In this algorithm, we use fluorescent nuclei staining to reproduce the hematoxylin information and natural tissue autofluorescence to reproduce the eosin information avoiding the necessity to specifically stain the proteins or intracellular structures with an additional fluorescence stain. Statistical Analysis Used: Our method is based on optimizing a transform function from fluorescence to H&E images using least mean square optimization. Results: It results in high quality virtual H&E digital images that can easily and efficiently be analyzed by pathologists. We validated our results with pathologists by making them annotate tumor in real and virtual H&E whole slide images and we obtained promising results. Conclusions: Hence, we provide a solution that enables pathologists to assess tissue and annotate specific structures
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International Nuclear Information System (INIS)
Latinovic, Olga; Reitz, Marvin; Le, Nhut M.; Foulke, James S.; Faetkenheuer, Gerd; Lehmann, Clara; Redfield, Robert R.; Heredia, Alonso
2011-01-01
R5 HIV-1 strains resistant to the CCR5 antagonist Maraviroc (MVC) can use drug-bound CCR5. We demonstrate that MVC-resistant HIV-1 exhibits delayed kinetics of coreceptor engagement and fusion during drug-bound versus free CCR5 infection of cell lines. Antibodies directed against the second extracellular loop (ECL2) of CCR5 had greater antiviral activity against MVC-bound compared to MVC-free CCR5 infection. However, in PBMCs, only ECL2 CCR5 antibodies HGS004 and HGS101, but not 2D7, inhibited infection by MVC resistant HIV-1 more potently with MVC-bound than with free CCR5. In addition, HGS004 and HGS101, but not 2D7, restored the antiviral activity of MVC against resistant virus in PBMCs. In flow cytometric studies, CCR5 binding by the HGS mAbs, but not by 2D7, was increased when PBMCs were treated with MVC, suggesting MVC increases exposure of the relevant epitope. Thus, HGS004 and HGS101 have antiviral mechanisms distinct from 2D7 and could help overcome MVC resistance.
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Whole blood flow cytometric analysis of Ureaplasma-stimulated monocytes from pregnant women.
Friedland, Yael D; Lee-Pullen, Tracey F; Nathan, Elizabeth; Watts, Rory; Keelan, Jeffrey A; Payne, Matthew S; Ireland, Demelza J
2015-06-01
We hypothesised that circulating monocytes of women with vaginal colonisation with Ureaplasma spp., genital microorganisms known to cause inflammation-driven preterm birth, would elicit a tolerised cytokine response to subsequent in vitro Ureaplasma parvum serovar 3 (UpSV3) stimulation. Using multi-parameter flow cytometry, we found no differences with regard to maternal colonisation status in the frequency of TNF-α-, IL-6-, IL-8- and IL-1β-expressing monocytes in response to subsequent UpSV3 stimulation (P > 0.10 for all cytokines). We conclude that vaginal Ureaplasma spp. colonisation does not specifically tolerise monocytes of pregnant women towards decreased responses to subsequent stimulation. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.
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Aggregate formation of eosin-Y adsorbed on nanocrystalline TiO2 films
Yaguchi, Kaori; Furube, Akihiro; Katoh, Ryuzi
2012-11-01
We have studied the adsorption of eosin-Y on nanocrystalline TiO2 films with two different solvents namely acetonitrile (ACN) and ethanol (EtOH). A Langmuir-type adsorption isotherm was observed with ACN. In contrast, a Freundlich-type adsorption isotherm was observed with EtOH, suggesting that EtOH molecules co-adsorbed on TiO2 surface. Absorption spectra of the dye adsorbed films clearly show aggregate formation at high concentrations of dye in the solutions. From the analysis of the spectra, we conclude that head-to-tail type aggregates are observed with ACN, whereas various types of aggregates, including H-type and head-to-tail type aggregates, are observed with EtOH.
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Silver nanoparticles plasmonic effect on eosin and rhodamine 6G luminescence in various media
Samusev, Ilia G.; Tikhomirova, Nadezhda S.; Slezhkin, Vasiliy A.; Zyubin, Andrey Yu.; Bryukhanov, Valery V.; Tsibulnikova, Anna V.
2016-11-01
The plasmonic enhancement and quenching of phosphorescence and fluorescence of the anionic (eosin) and cationic (rhodamine 6G) dyes have been studied in various environments: silver nanoparticles of silver hydrosol citrate in water, in polymer films and on the surface of nanoporous silica in order to determine the kinetic and spectral effects on the dye luminescence. Depending on the silver nanoparticles concentration both the enhancement and quenching of the dyes phosphorescence and fluorescence have been detected. The mechanism of interaction between the excited molecules and silver nanoparticles has been discussed.
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Directory of Open Access Journals (Sweden)
Kostić Ivana T.
2015-01-01
Full Text Available Despite the fact that the methods based on the osmotic properties of the cells are the most widely used for loading of drugs in human and animal erythrocytes, data related to the osmotic properties of erythrocytes derived from animal blood are scarce. This work was performed with an aim to investigate the possibility of use the flow cytometry as a tool for determination the osmotic behaviour of porcine and bovine erythrocytes, and thus facilitate the engineering of erythrocytes from animal blood to be drug carriers. The method of flow cytometry successfully provided the information about bovine and porcine erythrocyte osmotic fragility, and made the initial steps in assessment of erythrocyte shape in a large number of erythrocytes. Although this method is not able to confirm the swelling of pig erythrocytes, it indicated to the differences in pig erythrocytes that had basic hematological parameters inside and outside the reference values. In order to apply/use the porcine and bovine erythrocytes as drug carriers, the method of flow cytometry, confirming the presence of osmotically different fractions of red blood cells, indicated that various amounts of the encapsulated drug in porcine and bovine erythrocytes can be expected.
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WNT5A inhibits human dental papilla cell proliferation and migration
International Nuclear Information System (INIS)
Peng, L.; Ye, L.; Dong, G.; Ren, L.B.; Wang, C.L.; Xu, P.; Zhou, X.D.
2009-01-01
WNT proteins are a large family of cysteine-rich secreted molecules that are linked to both canonical and non-canonical signal pathways, and have been implicated in oncogenesis and tissue development. Canonical WNT proteins have been proven to play critical roles in tooth development, while little is known about the role of non-canonical WNT proteins such as WNT5A. In this study, WNT5A was localized to human dental papilla tissue and human dental papilla cells (HDPCs) cultured in vitro, using immunochemistry and RT-PCR. Recombinant adenovirus encoding full-length Wnt5a cDNA was constructed to investigate the biological role of WNT5A on HDPCs. The BrdU incorporation assay, the MTT assay and flow cytometric analysis showed that over-expression of Wnt5a strongly inhibited the proliferation of HDPCs in vitro. Wound healing and transwell migration assays indicated that over-expression of WNT5A reduced migration of HDPCs. In conclusion, our results showed that WNT5A negatively regulates both proliferation and migration of HDPCs, suggesting its important role in odontogenesis via controlling the HDPCs.
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International Nuclear Information System (INIS)
Ginting, Riski Titian; Yap, Chi Chin; Yahaya, Muhammad; Salleh, Muhamad Mat
2013-01-01
The influence of poly(2-methoxy-5-(2’-ethyl)-hexyloxy-p-phenylene vinylene) (MEH-PPV) and (6,6)-phenyl C61 butyric acid methyl ester (PCBM) weight ratio on the photovoltaic performance of inverted type organic solar cell based on Eosin-Y-coated ZnO nanorods has been investigated. Experimental results showed that the photovoltaic performance improved with weight ratio of MEH-PPV:PCBM from 1:1 to 1:3 due to better percolation pathway for electron transport and enhanced infiltration of polymer blend into interspace of Eosin-Y-coated ZnO nanorods. However, the overall performance started to decrease at weight ratio of 1:4 due to the aggregation of PCBM clusters which results in poor polymer blend infiltration. The optimum device at weight ratio of 1:3 exhibited short circuit current density of 3.95 ± 0.10 mA cm −2 , open circuit voltage of 0.53 ± 0.03 V, fill factor of 0.50 ± 0.03, and power conversion efficiency of 1.02 ± 0.07 %. - Highlights: • The device performance increased with donor:acceptor weight ratio up to 1:3. • Aggregation of fullerene-derivative led to poor infiltration at weight ratio of 1:4. • The optimum weight ratio was different from that of conventional device
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Energy Technology Data Exchange (ETDEWEB)
Ginting, Riski Titian [School of Applied Physics, Faculty of Science and Technology, Universiti Kebangsaan Malaysia, 43600 UKM Bangi, Selangor (Malaysia); Yap, Chi Chin, E-mail: ccyap@ukm.my [School of Applied Physics, Faculty of Science and Technology, Universiti Kebangsaan Malaysia, 43600 UKM Bangi, Selangor (Malaysia); Yahaya, Muhammad [School of Applied Physics, Faculty of Science and Technology, Universiti Kebangsaan Malaysia, 43600 UKM Bangi, Selangor (Malaysia); Salleh, Muhamad Mat [Institute of Microengineering and Nanoelectronics (IMEN), Universiti Kebangsaan Malaysia, 43600 UKM Bangi, Selangor (Malaysia)
2013-06-01
The influence of poly(2-methoxy-5-(2’-ethyl)-hexyloxy-p-phenylene vinylene) (MEH-PPV) and (6,6)-phenyl C61 butyric acid methyl ester (PCBM) weight ratio on the photovoltaic performance of inverted type organic solar cell based on Eosin-Y-coated ZnO nanorods has been investigated. Experimental results showed that the photovoltaic performance improved with weight ratio of MEH-PPV:PCBM from 1:1 to 1:3 due to better percolation pathway for electron transport and enhanced infiltration of polymer blend into interspace of Eosin-Y-coated ZnO nanorods. However, the overall performance started to decrease at weight ratio of 1:4 due to the aggregation of PCBM clusters which results in poor polymer blend infiltration. The optimum device at weight ratio of 1:3 exhibited short circuit current density of 3.95 ± 0.10 mA cm{sup −2}, open circuit voltage of 0.53 ± 0.03 V, fill factor of 0.50 ± 0.03, and power conversion efficiency of 1.02 ± 0.07 %. - Highlights: • The device performance increased with donor:acceptor weight ratio up to 1:3. • Aggregation of fullerene-derivative led to poor infiltration at weight ratio of 1:4. • The optimum weight ratio was different from that of conventional device.
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Energy Technology Data Exchange (ETDEWEB)
Zhou, Gang [Nanjing University, State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences (China); Liu, Naicheng; Wang, Zhenheng [Nanjing University, Department of Orthopedics, Jinling Hospital, School of Medicine (China); Shi, Tongguo; Gan, Jingjing; Wang, Zhenzhen; Zhang, Junfeng, E-mail: jfzhang@nju.edu.cn [Nanjing University, State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences (China)
2017-02-15
Nanoparticle-based applications for diagnostics and therapeutics have been extensively studied. These applications require a profound understanding of the fate of nanoparticles (NPs) in cellular environments. However, until now, few analytical methods are available and most of them rely on fluorescent properties or special elements of NPs; therefore, for NPs without observable optical properties or special elements, the existing methods are hardly applicable. In this study, we introduce a flow cytometry light scattering (FCLS)-based approach that quantifies in situ NPs accurately in mammalian cells. Continuous cells of heterogeneous human epithelial colorectal adenocarcinoma (Caco-2 cells), mouse peritoneal macrophages (MPM), and human adenocarcinomic alveolar basal epithelia (A549 cells) were cultured with NPs with certain concentrations and size. The intensity of the flow cytometric side scattered light, which indicates the quantity of NPs in the cells, was analyzed. The result shows an accurate size- and dose-dependent uptake of Au NPs (5, 30, 250 nm) in Caco-2 cells. The size- and dose- dependence of Au NPs (5, 30, 250 nm) and carbon NPs (50, 500 nm) in cells was validated by transmission electron microscope (TEM). This paper demonstrates the great potential of flow cytometry light scattering in the quantitative study of the size and dose effect on in situ metallic or non-metallic NPs in mammalian cells.
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Zhou, Gang; Liu, Naicheng; Wang, Zhenheng; Shi, Tongguo; Gan, Jingjing; Wang, Zhenzhen; Zhang, Junfeng
2017-02-01
Nanoparticle-based applications for diagnostics and therapeutics have been extensively studied. These applications require a profound understanding of the fate of nanoparticles (NPs) in cellular environments. However, until now, few analytical methods are available and most of them rely on fluorescent properties or special elements of NPs; therefore, for NPs without observable optical properties or special elements, the existing methods are hardly applicable. In this study, we introduce a flow cytometry light scattering (FCLS)-based approach that quantifies in situ NPs accurately in mammalian cells. Continuous cells of heterogeneous human epithelial colorectal adenocarcinoma (Caco-2 cells), mouse peritoneal macrophages (MPM), and human adenocarcinomic alveolar basal epithelia (A549 cells) were cultured with NPs with certain concentrations and size. The intensity of the flow cytometric side scattered light, which indicates the quantity of NPs in the cells, was analyzed. The result shows an accurate size- and dose-dependent uptake of Au NPs (5, 30, 250 nm) in Caco-2 cells. The size- and dose- dependence of Au NPs (5, 30, 250 nm) and carbon NPs (50, 500 nm) in cells was validated by transmission electron microscope (TEM). This paper demonstrates the great potential of flow cytometry light scattering in the quantitative study of the size and dose effect on in situ metallic or non-metallic NPs in mammalian cells.
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Exfoliated graphite/titanium dioxide nanocomposites for photodegradation of eosin yellow
Ndlovu, Thabile; Kuvarega, Alex T.; Arotiba, Omotayo A.; Sampath, Srinivasan; Krause, Rui W.; Mamba, Bhekie B.
2014-05-01
An improved photocatalyst consisting of a nanocomposite of exfoliated graphite and titanium dioxide (EG-TiO2) was prepared. SEM and TEM micrographs showed that the spherical TiO2 nanoparticles were evenly distributed on the surface of the EG sheets. A four times photocatalytic enhancement was observed for this floating nanocomposite compared to TiO2 and EG alone for the degradation of eosin yellow. For all the materials, the reactions followed first order kinetics where for EG-TiO2, the rate constant was much higher than for EG and TiO2 under visible light irradiation. The enhanced photocatalytic activity of EG-TiO2 was ascribed to the capability of graphitic layers to accept and transport electrons from the excited TiO2, promoting charge separation. This indicates that carbon, a cheap and abundant material, can be a good candidate as an electron attracting reservoir for photocatalytic organic pollutant degradation.
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5th International Conference on Jets, Wakes and Separated Flows
2016-01-01
This volume collects various contributions from the 5th International Conference on Jets, Wakes and Separated Flows (ICJWSF2015) that took place in Stockholm during June 2015. Researchers from all around the world presented their latest results concerning fundamental and applied aspects of fluid dynamics. With its general character, the conference embraced many aspects of fluid dynamics, such as shear flows, multiphase flows and vortex flows, for instance. The structure of the present book reflects the variety of topics treated within the conference i.e. Jets, Wakes, Separated flows, Vehicle aerodynamics, Wall-bounded and confined flows, Noise, Turbomachinery flows, Multiphase and reacting flows, Vortex dynamics, Energy-related flows and a section dedicated to Numerical analyses.
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Tanev, Stoyan; Sun, Wenbo; Pond, James; Tuchin, Valery V; Zharov, Vladimir P
2009-09-01
The formulation of the finite-difference time-domain (FDTD) approach is presented in the framework of its potential applications to in-vivo flow cytometry based on light scattering. The consideration is focused on comparison of light scattering by a single biological cell alone in controlled refractive-index matching conditions and by cells labeled by gold nanoparticles. The optical schematics including phase contrast (OPCM) microscopy as a prospective modality for in-vivo flow cytometry is also analyzed. The validation of the FDTD approach for the simulation of flow cytometry may open up a new avenue in the development of advanced cytometric techniques based on scattering effects from nanoscale targets. 2009 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
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Energy Technology Data Exchange (ETDEWEB)
Hosono, Eiji; Fujihara, Shinobu; Kimura, Toshio
2004-06-15
Micro/nano-textured ZnO thick films were synthesized through deposition and pyrolysis of layered hydroxide zinc acetate (LHZA), Zn{sub 5}(OH){sub 8}(CH{sub 3}COO){sub 2}{center_dot}2H{sub 2}O. LHZA films having a unique, rose-like morphology were initially deposited on conducting glass sheets in a chemical bath composed of methanol and zinc acetate dihydrate at 60 deg. C under neutral conditions. Pyrolysis of the LHZA films resulted in formation of ZnO without destroying the original morphology. Pyrolysis temperatures were found to greatly influence grain sizes and specific surface areas of the ZnO films. Photoelectrochemical performance of the films as ZnO/eosin Y electrodes was investigated in dye-sensitized solar cells using an I{sup -}/I{sub 3}{sup -} redox electrolyte solution. The cell using the ZnO film pyrolyzed at 150 deg. C exhibited overall light to electricity conversion efficiencies of 2.0 and 3.3% under an AM-1.5 illumination at 100 and 10 mW cm{sup -2}, respectively. While microscale pores in the electrodes facilitated mass transfer of fluid electrolytes in the depth direction, nanoscale pores contributed to an increase in the amount of adsorbed dye. The maximum incident photon-to-current conversion efficiency (IPCE) of the electrode reached 84.9% at a wavelength of 530 nm.
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Flow cytometric monitoring of influenza A virus infection in MDCK cells during vaccine production
Directory of Open Access Journals (Sweden)
Reichl Udo
2008-04-01
Full Text Available Abstract Background In cell culture-based influenza vaccine production the monitoring of virus titres and cell physiology during infection is of great importance for process characterisation and optimisation. While conventional virus quantification methods give only virus titres in the culture broth, data obtained by fluorescence labelling of intracellular virus proteins provide additional information on infection dynamics. Flow cytometry represents a valuable tool to investigate the influences of cultivation conditions and process variations on virus replication and virus yields. Results In this study, fluorescein-labelled monoclonal antibodies against influenza A virus matrix protein 1 and nucleoprotein were used for monitoring the infection status of adherent Madin-Darby canine kidney cells from bioreactor samples. Monoclonal antibody binding was shown for influenza A virus strains of different subtypes (H1N1, H1N2, H3N8 and host specificity (human, equine, swine. At high multiplicity of infection in a bioreactor, the onset of viral protein accumulation in adherent cells on microcarriers was detected at about 2 to 4 h post infection by flow cytometry. In contrast, a significant increase in titre by hemagglutination assay was detected at the earliest 4 to 6 h post infection. Conclusion It is shown that flow cytometry is a sensitive and robust method for the monitoring of viral infection in fixed cells from bioreactor samples. Therefore, it is a valuable addition to other detection methods of influenza virus infection such as immunotitration and RNA hybridisation. Thousands of individual cells are measured per sample. Thus, the presented method is believed to be quite independent of the concentration of infected cells (multiplicity of infection and total cell concentration in bioreactors. This allows to perform detailed studies on factors relevant for optimization of virus yields in cell cultures. The method could also be used for process
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Hirschsprung′s disease diagnosis: Comparison of immunohistochemical, hematoxilin and eosin staining
Directory of Open Access Journals (Sweden)
Memarzadeh Mehrdad
2009-01-01
Full Text Available Background : The diagnosis of Hirschsprung′s disease (HD is based on the absence of ganglion cells. In hemotoxilin and eosin (H and E as well as acetylcholine esterase staining there are limitations in the diagnosis of immature ganglion cells in neonates. Methods : In this prospective study, 54 biopsies taken from suspected HD patients (five mucosal specimens and 49 full thickness specimens were studied. In the laboratory, after preparing sections of paraffin embedded tissues, H and E staining slides were compared with immunohistochemical (IHC staining including: S100, NSE, CD117, CD56, Cathepsin D, Vimentin, BCL2, GFAP, Synaptophysin and chromogranin. Results : The study revealed 30 negative (absence of ganglion cells cases (55.5%, 17 positive cases (31.04% and seven suspected cases (12.9% of ganglion cells on the H and E staining. On IHC staining with CD56 and Cathepsin D, all of the 17 positive cases detected through H and E, were confirmed for having ganglion cells and out of 30 cases reported negative on H and E staining, 28(93.3% were reported negative and two (6.7% positive by IHC staining. Of the seven suspected cases H and E staining, IHC staining detectedganglion cells only in five slides; two remained negative. Conclusions : IHC staining using CD56 and Cathepsin D improved the accuracy of diagnosis in HD when used in addition to H and E staining technique, especially for negative or suspicious slides.
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Analysis of the Budding Yeast Cell Cycle by Flow Cytometry.
Rosebrock, Adam P
2017-01-03
DNA synthesis is one of the landmark events in the cell cycle: G 1 cells have one copy of the genome, S phase cells are actively engaged in DNA synthesis, and G 2 cells have twice as much nuclear DNA as G 1 cells. Cellular DNA content can be measured by staining with a fluorescent dye followed by a flow-cytometric readout. This method provides a quantitative measurement of cell cycle position on a cell-by-cell basis at high speed. Using flow cytometry, tens of thousands of single-cell measurements can be generated in a few seconds. This protocol details staining of cells of the budding yeast Saccharomyces cerevisiae for flow cytometry using Sytox Green dye in a method that can be scaled widely-from one sample to many thousands and operating on inputs ranging from 1 million to more than 100 million cells. Flow cytometry is preferred over light microscopy or Coulter analyses for the analysis of the cell cycle as DNA content and cell cycle position are being directly measured. © 2017 Cold Spring Harbor Laboratory Press.
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Kalina, Tomas; Flores-Montero, Juan; Lecrevisse, Quentin; Pedreira, Carlos E; van der Velden, Vincent H J; Novakova, Michaela; Mejstrikova, Ester; Hrusak, Ondrej; Böttcher, Sebastian; Karsch, Dennis; Sędek, Łukasz; Trinquand, Amelie; Boeckx, Nancy; Caetano, Joana; Asnafi, Vahid; Lucio, Paulo; Lima, Margarida; Helena Santos, Ana; Bonaccorso, Paola; van der Sluijs-Gelling, Alita J; Langerak, Anton W; Martin-Ayuso, Marta; Szczepański, Tomasz; van Dongen, Jacques J M; Orfao, Alberto
2015-02-01
Flow cytometric immunophenotyping has become essential for accurate diagnosis, classification, and disease monitoring in hemato-oncology. The EuroFlow Consortium has established a fully standardized "all-in-one" pipeline consisting of standardized instrument settings, reagent panels, and sample preparation protocols and software for data analysis and disease classification. For its reproducible implementation, parallel development of a quality assurance (QA) program was required. Here, we report on the results of four consecutive annual rounds of the novel external QA EuroFlow program. The novel QA scheme aimed at monitoring the whole flow cytometric analysis process (cytometer setting, sample preparation, acquisition and analysis) by reading the median fluorescence intensities (MedFI) of defined lymphocytes' subsets. Each QA participant applied the predefined reagents' panel on blood cells of local healthy donors. A uniform gating strategy was applied to define lymphocyte subsets and to read MedFI values per marker. The MedFI values were compared with reference data and deviations from reference values were quantified using performance score metrics. In four annual QA rounds, we analyzed 123 blood samples from local healthy donors on 14 different instruments in 11 laboratories from nine European countries. The immunophenotype of defined cellular subsets appeared sufficiently standardized to permit unified (software) data analysis. The coefficient of variation of MedFI for 7 of 11 markers performed repeatedly below 30%, average MedFI in each QA round ranged from 86 to 125% from overall median. Calculation of performance scores was instrumental to pinpoint standardization failures and their causes. Overall, the new EuroFlow QA system for the first time allowed to quantify the technical variation that is introduced in the measurement of fluorescence intensities in a multicentric setting over an extended period of time. EuroFlow QA is a proficiency test specific for
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Huang, Binbin; Zhao, Yating; Yang, Chao; Gao, Yuan; Xia, Wujiong
2017-07-21
A mild, metal-free, and absolutely para-selective bromination of aniline derivatives has been developed in excellent yields, wherein the organic dye Eosin Y is employed as the bromine source in company with Selectfluor. Neither air nor moisture sensitive, this facile reaction proceeds smoothly at room temperature and completes within a short time. Mechanistic studies indicate a radical pathway; therefore, the existence of an in situ generated brominating reagent, "Selectbrom", is postulated, which may reasonably account for the unique regioselectivity for the para-bromination of N-acyl- as well as N-sulfonylanilines.
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Design of pneumatic proportional flow valve type 5/3
Laski, P. A.; Pietrala, D. S.; Zwierzchowski, J.; Czarnogorski, K.
2017-08-01
In this paper the 5/3-way pneumatic, proportional flow valve was designed and made. Stepper linear actuator was used to move the spool. The valve is controlled by the controlled based on a AVR microcontroller. Virtual model of the valve was created in CAD. The real element was made based on a standard 5/3-way manually actuated valve with hand lever, which was dismounted and replaced by linear stepper motor. All the elements was mounted in a specially made housing. The controller consists of microcontroller Atmega16, integrated circuit L293D, display, two potentiometers, three LEDs and six buttons. Series of research was also conducted. Simulation research were performed using CFD by the Flow Simulation addition to SolidWorks. During the experiments the valve characteristics of flow and pressure was determined.
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Comparison of special stains for keratin with routine hematoxylin and eosin stain.
Rao, Roopa S; Patil, Shankargouda; Majumdar, Barnali; Oswal, Rakesh G
2015-03-01
Keratins are the most abundant proteins and are characteristic findings in many epithelial pathologies, making it a diagnostically important marker, both histopathologically and immunohistochemically. Since, immunohistochemistry is an expensive diagnostic tool, special stains to detect the degree of keratinization could serve as a faster and economic option. The aim of the present study was to compare the efficacy of special stains for keratin with standard hematoxylin and eosin stain (H and E). Objectives include: (i) To subject the diagnosed cases of keratin disorders to the selected special stains: Ayoub-shklar method, Dane-Herman method, Alcian blue -periodic acid Schiff 's (PAS), rapid papanicolaou (PAP) and Gram's stain. (ii) To compare the staining specificity and staining intensity of special stains with respect to routine hematoxylin and eosin (H and E) stain. (iii) To compare the efficacy of special stains to routine H and E stain in identification of the type of keratin present in the selected cases. A total of 80 cases of known pathology for keratin were retrieved from the department archive, which included 10 each of normal gingiva, hyperkeratosis, squamous papilloma, verrucous hyperplasia, verrucous carcinoma, well-differentiated squamous cell carcinoma, orthokeratinized odontogenic cyst and keratocystic odontogenic tumors. Six sections of 4 µ each from the paraffin blocks were made, stained with H and E and the special stains and these were evaluated by 2 pathologists based on the modified scoring criteria from Rahma Al-Maaini and Philip Bryant 2008. The results were tabulated using Chi square and kappa statistics. The statistical values for identification of the type of keratinization was insignificant showing that ortho and parakeratinized epithelia could be correctly identified by both H and E as well as all the special stains. Furthermore, all the special stains showed a positive result and statistical significance (P < 0.001) with respect to
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Directory of Open Access Journals (Sweden)
Mohamed Y. Dhamra
2014-06-01
Full Text Available A simple, sensitive and rapid spectrophotometric method was developed and validated for the determination of terbutaline sulphate and tetracycline hydrochloride drugs in pure form and pharmaceutical formulations. The proposed method is based on the formation of binary complexes between these drugs and eosin Y in aqueous acetate buffered medium. Under the optimum conditions, the binary complexes showed absorption maxima at 545 nm. Beer's law was rectilinear over concentration range of 0.5-10 and 5-45 μg/mL, R2 were 0.9984 and 0.9988, RSD were ≤ 0.72 and ≤ 0.19 (n=5 with average recovery % 101.42 % and 100.08 % and the average recovery values of pharmaceutical formulations 101.48 and 98.01 for above drugs respectively. The limit of detection (LOD were 0.030 and 0.613 μg/mL and limit of quantitation (LOQ were 0.103 and 2.00 μg/mL with molar absorptivity values 3.169 103 and 6.347 103 l. mol-1. cm-1 and the relative standard deviation values ≤0.720 and ≤ 0.19 for both drugs respectively. No interference was observed from the excipients that are commonly present in pharmaceutical formulations. The proposed method was successfully applied to the analysis of terbutaline sulphate tablet and tetracycline hydrochloride capsule in their dosage forms.
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International Nuclear Information System (INIS)
Dhamra, M.Y.; Sabha, T.N.A.; Ghabsha, T.S.A.
2014-01-01
A simple, sensitive and rapid spectrophotometric method was developed and validated for the determination of terbutaline sulphate and tetracycline hydrochloride drugs in pure form and pharmaceutical formulations. The proposed method is based on the formation of binary complexes between these drugs and eosin Y in aqueous acetate buffered medium. Under the optimum conditions, the binary complexes showed absorption maxima at 545 nm. Beer's law was rectilinear over concentration range of 0.5-10 and 5-45 micro g/mL, R/sub 2/ were 0.9984 and 0.9988, RSD were 0.72 and 0.19 (n=5) with average recovery 101.42 % and 100.08 % and the average recovery values of pharmaceutical formulations 101.48 and 98.01 for above drugs respectively. The limit of detection (LOD) were 0.030 and 0.613 micro g/mL and limit of quantitation (LOQ) were 0.103 and 2.00 micro g/mL with molar absorptivity values 3.169 10/sup 3/ and 6.347 10/sup 3/l. mol/sup -1/. Cm/sup -1/ and the relative standard deviation values 0.720 and 0.19 for both drugs respectively. No interference was observed from the excipients that are commonly present in pharmaceutical formulations. The proposed method was successfully applied to the analysis of terbutaline sulphate tablet and tetracycline hydrochloride capsule in their dosage forms. (author)
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Acierno, M M; Raj, K; Giger, U
2014-01-01
The Dog erythrocyte antigen (DEA) 1 blood group system was thought to contain types DEA 1.1 and 1.2 (and possibly 1.3 [A3]). However, DEA 1.2+ dogs are very rare and newer typing methods reveal varying degrees of DEA 1 positivity. To assess if variation in DEA 1 positivity is because of quantitative differences in surface antigen expression. To determine expression patterns in dogs over time and effects of blood storage (4°C). To evaluate DEA 1.2+ samples by DEA 1 typing methods. Anticoagulated blood samples from 66 dogs in a research colony and from a hospital, and 9 previously typed DEA 1.2+ dogs from an animal blood bank. Research study: Samples were analyzed by flow cytometry and immunochromatographic strip using a monoclonal anti-DEA 1 antibody. Twenty dogs were DEA 1-, whereas 46 dogs were weakly to strongly DEA 1+. Antigen quantification revealed excellent correlation between strip and flow cytometry (r = 0.929). Both methods reclassified DEA 1.2+ samples as weakly to moderately DEA 1+, but they were not retyped with the polyclonal anti-DEA 1.1/1.X antibodies. Dogs and blood samples retained their relative DEA 1 antigen densities over time. The blood group system DEA 1 is a continuum from negative to strongly positive antigen expression. Previously typed DEA 1.2+ appears to be DEA 1+. These findings further the understanding of the DEA 1 system and suggest that all alleles within the DEA 1 system have a similarly based epitope recognized by the monoclonal antibody. Copyright © 2014 by the American College of Veterinary Internal Medicine.
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Quick cytogenetic screening of breeding bulls using flow cytometric sperm DNA histogram analysis.
Nagy, Szabolcs; Polgár, Péter J; Andersson, Magnus; Kovács, András
2016-09-01
The aim of the present study was to test the FXCycle PI/RNase kit for routine DNA analyses in order to detect breeding bulls and/or insemination doses carrying cytogenetic aberrations. In a series of experiments we first established basic DNA histogram parameters of cytogenetically healthy breeding bulls by measuring the intraspecific genome size variation of three animals, then we compared the histogram profiles of bulls carrying cytogenetic defects to the baseline values. With the exception of one case the test was able to identify bulls with cytogenetic defects. Therefore, we conclude that the assay could be incorporated into the laboratory routine where flow cytometry is applied for semen quality control.
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Katarzyna Czaczyk; Wojciech Juzwa
2012-01-01
Background. Flow cytometry constitutes an alternative for traditional methods of microorganisms identifi cation and analysis, including methods requiring cultivation step. It enables the detection of pathogens and other microorganisms contaminants without the need to culture microbial cells meaning that the sample (water, waste or food e.g. milk, wine, beer) may be analysed directly. This leads to a signifi cant reduction of time required for analysis allowing monitoring of production process...
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Pavón, Natalia; Correa, Francisco; Buelna-Chontal, Mabel; Hernández-Esquivel, Luz; Chávez, Edmundo
2015-10-15
Mitochondrial permeability transition is a process established through massive Ca(2+) load in addition to an inducer reagent. Ebselen (Ebs), an antioxidant seleno compound, has been introduced as a reagent which inhibits mitochondrial dysfunction induced by permeability transition. Paradoxically enough, it has been shown that Ebs may also be able to induce the opening of the mitochondrial non-selective pores. This study was performed with the purpose of establishing the membrane system involved in Ebs-induced pore opening. Permeability transition was appraised by analyzing the following: i) matrix Ca(2+) release, and mitochondrial swelling, ii) efflux of cytochrome c, and iii) the inhibition of superoxide dismutase. All of these adverse reactions were inhibited by N-ethylmaleimide and cyclosporin A. At concentrations from 5 to 20 μM, we found that Ebs induces non-specific membrane permeability. Remarkably, Ebs blocks the binding of the fluorescent reagent eosin-5-maleimide to the thiol groups of the adenine nucleotide translocase. Based on the above, it is tempting to hypothesize that Ebs induces pore opening through its binding to the ADP/ATP carrier. Copyright © 2015 Elsevier Inc. All rights reserved.
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Multiphase Flow Dynamics 5 Nuclear Thermal Hydraulics
Kolev, Nikolay Ivanov
2012-01-01
The present Volume 5 of the successful book package "Multiphase Flow Dynamics" is devoted to nuclear thermal hydraulics which is a substantial part of nuclear reactor safety. It provides knowledge and mathematical tools for adequate description of the process of transferring the fission heat released in materials due to nuclear reactions into its environment. It step by step introduces into the heat release inside the fuel, temperature fields in the fuels, the "simple" boiling flow in a pipe described using ideas of different complexity like equilibrium, non equilibrium, homogeneity, non homogeneity. Then the "simple" three-fluid boiling flow in a pipe is described by gradually involving the mechanisms like entrainment and deposition, dynamic fragmentation, collisions, coalescence, turbulence. All heat transfer mechanisms are introduced gradually discussing their uncertainty. Different techniques are introduced like boundary layer treatments or integral methods. Comparisons with experimental data at each step...
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Multiphase flow dynamics 5 nuclear thermal hydraulics
Kolev, Nikolay Ivanov
2015-01-01
This Volume 5 of the successful book package "Multiphase Flow Dynamics" is devoted to nuclear thermal hydraulics which is a substantial part of nuclear reactor safety. It provides knowledge and mathematical tools for adequate description of the process of transferring the fission heat released in materials due to nuclear reactions into its environment. It step by step introduces into the heat release inside the fuel, temperature fields in the fuels, the "simple" boiling flow in a pipe described using ideas of different complexity like equilibrium, non equilibrium, homogeneity, non homogeneity. Then the "simple" three-fluid boiling flow in a pipe is described by gradually involving the mechanisms like entrainment and deposition, dynamic fragmentation, collisions, coalescence, turbulence. All heat transfer mechanisms are introduced gradually discussing their uncertainty. Different techniques are introduced like boundary layer treatments or integral methods. Comparisons with experimental data at each step demons...
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The effect of 0.5% ropivacaine on epidural blood flow
DEFF Research Database (Denmark)
Dahl, J B; Simonsen, L; Mogensen, T
1990-01-01
Twenty patients scheduled for elective abdominal surgery received epidural analgesia with 20 ml 0.5% ropivacaine or 0.5% bupivacaine. Epidural blood flow was measured by an epidural 133Xe clearance technique on the day before surgery (no local anaesthetic) and again 1 h before surgery, 30 min after...... injection of the local anaesthetic during continuous infusion (8 ml/h). Median initial blood flow was 5.0 ml/min and 6.0 ml/min per 100 g tissue in patients receiving ropivacaine and bupivacaine, respectively. After epidural bupivacaine, blood flow increased in 8 of 10 patients to 6.9 ml/min per 100 g...... tissue (P less than 0.05) in contrast to a decrease in 9 of 10 patients to 3.3 ml/min per 100 g tissue after ropivacaine (P less than 0.05), (P less than 0.01 between groups). The median level of sensory analgesia was T3.5 and T4.5 in the ropivacaine and bupivacaine group, respectively (P greater than 0...
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International Nuclear Information System (INIS)
Liu Ping; Guan Qingbao; Gu Aijuan; Liang Guozheng; Yuan Li; Chang Jianfei
2011-01-01
Interface is Key topic of developing advanced fiber reinforced polymeric composites. Novel advanced glass woven fabric (GF) reinforced composites, coded as GF/mBT, were prepared, of which the matrix resin was hyperbranched polysiloxane (HBPSi) modified maleimide-triazine (mBT) resin. The influence of the composition of the matrix on the interfacial nature of the GF/mBT composites were studied and compared with that of the composite based on GF and BT resin using contact angle, X-ray photoelectron spectroscopy (XPS), scanning electron microscope (SEM), and dielectric properties over wide frequency and temperature ranges. Results show that the interfacial nature of the composites is dependent on the chemistries of the matrices, mBT matrices have better interfacial adhesion with GF than BT resin owing to the formation of chemical and hydrogen bonds between mBT resin and GF; while in the case of mBT resins, the content of HBPSi also plays an important role on the interfacial feature and thus the macro-performance. Specifically, with increasing the content of HBPSi in the matrix, the interlaminate shear strength of corresponding composites significantly improves, demonstrating that better interfacial adhesion guarantees outstanding integrated properties of the resultant composites.
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Cerezo, Maria I.
2017-02-17
Polycyclic aromatic hydrocarbons are very important components of oil pollution. These pollutants tend to sorb to cell surfaces, exerting toxic effects on organisms. Our study developed a flow cytometric method for the detection of PAHs sorbed to phytoplankton by exploiting their spectral characteristics. We discriminated between cells with PAHs from cells free of PAHs. Clear discrimination was observed with flow cytometer provided with 375 or 405nm lasers in addition to the standard 488nm laser necessary to identify phytoplankton. Using this method, we measured the relationship between the percentages of phytoplankton organisms with PAHs, with the decrease in the growth rate. Moreover, the development of this method could be extended to facilitate the study of PAHs impact on cell cultures from a large variety of organisms.
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Myslitskaya, N. A.; Samusev, I. G.; Bryukhanov, V. V.
2014-11-01
The electron excitation energy transfer from CdSe/ZnS quantum dots to eosin molecules in the polymer matrix of butyral resin is investigated. The main characteristics of energy transfer are determined. By means of luminescence microscopy and correlation spectroscopy methods we found that quantum dots in the polymer are in an aggregate state.
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Energy Technology Data Exchange (ETDEWEB)
Jones, G. II; Farahat, C.W.; Oh, C. (Boston Univ., MA (United States))
1994-07-14
The electron-transfer photochemistry of the covalent derivatives of the dye eosin, in which the xanthene dye is covalently attached to the amino acid L-tryptophan via the thiohydantoin derivative, the tryptophan dipeptide, and an ethyl ester derivative, has been investigated. The singlet excited state of the dye is significantly quenched on attachment of the aromatic amino acid residue. Dye triplet states are also intercepted through intramolecular interaction of excited dye and amino acid pendants. Flash photolysis experiments verify that this interaction involves electron transfer from the indole side chains of tryptophan. Rate constants for electron transfer are discussed in terms of the distance relationships for the eosin chromophore and aromatic redox sites on peptide derivatives, the pathway for [sigma]-[pi] through-bond interaction between redox sites, and the multiplicity and state of protonation for electron-transfer intermediates. Selected electron-transfer photoreactions were studied under conditions of binding of the peptide derivatives in a high molecular weight, water-soluble, globular polymer, poly(vinyl-2-pyrrolidinone). 28 refs., 4 figs., 1 tab.
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Ratiometric fluorescence polarization as a cytometric functional parameter: theory and practice
International Nuclear Information System (INIS)
Yishai, Yitzhak; Fixler, Dror; Cohen-Kashi, Meir; Zurgil, Naomi; Deutsch, Mordechai
2003-01-01
The use of ratiometric fluorescence polarization (RFP) as a functional parameter in monitoring cellular activation is suggested, based on the physical phenomenon of fluorescence polarization dependency on emission wavelengths in multiple (at least binary) solutions. The theoretical basis of this dependency is thoroughly discussed and examined via simulation. For simulation, aimed to imitate a fluorophore-stained cell, real values of the fluorescence spectrum and polarization of different single fluorophore solutions were used. The simulation as well as the experimentally obtained values of RFP indicated the high sensitivity of this measure. Finally, the RFP parameter was utilized as a cytometric measure in three exemplary cellular bioassays. In the first, the apoptotic effect of oxLDL in a human Jurkat FDA-stained T cell line was monitored by RFP. In the second, the interaction between cell surface membrane receptors of human T lymphocyte cells was monitored by RFP measurements as a complementary means to the fluorescence resonance energy transfer (FRET) technique. In the third bioassay, cellular thiol level of FDA- and CMFDA-labelled Jurkat T cells was monitored via RFP
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Directory of Open Access Journals (Sweden)
Reginaldo Cipullo
2011-08-01
Full Text Available FUNDAMENTO: O significado clínico de vasculites, lesões isquêmicas, efeito Quilty e da presença de eosinófilos em biópsias endomiocárdicas de receptores de transplante cardíaco com rejeições leves não foi ainda estabelecido. OBJETIVO: Verificar se esses achados histológicos encontrados nas biópsias endomiocárdicas (eosinófilos, vasculites, efeito Quilty e lesões isquêmicas são capazes de predizer rejeição aguda do enxerto. MÉTODOS: Foram reavaliadas 1.012 biópsias endomiocárdicas consecutivas; dessas, 939 foram classificadas como OR ou 1R pela Nomenclatura da Sociedade Internacional de Transplante de Coração e Pulmão de 2005, e divididas em dois grupos: (1 Biópsias preditoras: aquelas que precederam rejeição aguda; e (2 Biópsias não preditoras: aquelas que não precederam rejeição aguda. Comparamos a ocorrência dos seguintes achados histológicos: vasculites, lesões isquêmicas, efeito Quilty e eosinófilos por análise uni e multivariada entre os grupos. RESULTADOS: Após análise estatística verificou-se a presença de vasculite intensa e de eosinófilos como maiores preditores para rejeição aguda futura, apresentando respectivamente as seguintes razões de chance: 10,60 (IC95%: 3,62 - 31,06. p FUNDAMENTO: El significado clínico de vasculitis, lesiones isquémicas, efecto Quilty y de la presencia de eosinófilos en biopsias endomiocárdicas de receptores de transplante cardíaco con rechazos leves no fue aun establecido. OBJETIVO: Verificar si esos hallazgos histológicos encontrados en las biopsias endomiocárdicas (eosinófilos, vasculitis, efecto Quilty y lesiones isquémicas son capaces de predecir rechazo agudo del injerto. MÉTODOS: Fueron reevaluadas 1.012 biopsias endomiocárdicas consecutivas; de esas, 939 fueron clasificadas como OR o 1R por la Nomenclatura de la Sociedad Internacional de Transplante de Corazón y Pulmón de 2005, y divididas en dos grupos: (1 Biopsias predictoras: aquellas que
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Oyelude, Emmanuel O; Awudza, Johannes A M; Twumasi, Sylvester K
2017-09-22
Low-cost teak leaf litter powder (TLLP) was prepared as possible substitute for activated carbon. The feasibility of using the adsorbent to remove eosin yellow (EY) dye from aqueous solution was investigated through equilibrium adsorption, kinetic and thermodynamic studies. The removal of dye from aqueous solution was feasible but influenced by temperature, pH, adsorbent dosage and contact time. Variation in the initial concentration of dye did not influence the equilibrium contact time. Optimum adsorption of dye occurred at low adsorbent dosages, alkaline pH and high temperatures. Langmuir isotherm model best fit the equilibrium adsorption data and the maximum monolayer capacity of the adsorbent was 31.64 mg g -1 at 303 K. The adsorption process was best described by pseudo-second order kinetic model at 303 K. Boundary layer diffusion played a key role in the adsorption process. The mechanism of uptake of EY by TLLP was controlled by both liquid film diffusion and intraparticle diffusion. The values of mean adsorption free energy, E (7.91 kJ mol -1 ), and standard enthalpy, ΔH° (+13.34 kJ mol -1 ), suggest physical adsorption. The adsorption process was endothermic and spontaneous. Teak leaf litter powder is a promising low-cost adsorbent for treating wastewaters containing eosin yellow.
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Flow cytometric detection of viruses in the Zuari estuary, Goa
Digital Repository Service at National Institute of Oceanography (India)
Mitbavkar, S.; Rajaneesh, K.M.; SathishKumar, P.
and virus-mediated processes for better understanding of the microbial food web and the biogeochemistry. 1. Suttle, C. A., Nature, 2005, 437, 356– 361. 2. Danovaro, R. et al., Freshwater Biol., 2008, 53, 1186–1213. 3. Suttle, C. A., Nature, 2007, 5... of the microbial food web, with abundance in marine waters ranging from 10 6 ml –1 in the deep sea to 10 8 ml –1 in coastal waters and 10 9 g –1 of dry weight in the marine sediments 1,2 , which is usually 15-fold greater than bacterial and archael...
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Fraser, Kathleen; Abbas, Mariam; Hull, Peter R
2014-01-01
We previously demonstrated that repeated intradermal steroid injections administered at weekly intervals into positive patch-test sites induce hyposensitization and desensitization. To examine changes in CD4CD25CD127lo/ regulatory T cells during the attenuation of the patch-test response. Ten patients with known allergic contact dermatitis were patch tested weekly for 10 weeks. The patch-test site was injected intradermally with 2 mg triamcinolone. At weeks 1 and 7, a biopsy was performed on the patch-test site in 6 patients, and flow cytometry was performed assessing CD4CD25CD127lo/ regulatory T cells. Secondary outcomes were clinical score, reaction size, erythema, and temperature. Statistical analysis included regression, correlation, and repeated-measures analysis of variance. The percentage of CD4CD25CD127lo/ regulatory T cells, measured by flow cytometry, increased from week 1 to week 7 by an average of 19.2%. The average grade of patch-test reaction decreased from +++ (vesicular reaction) to ++ (palpable erythema). The mean drop in temperature following treatment was 0.28°C per week. The mean area decreased 8.6 mm/wk over 10 weeks. Intradermal steroid injections of weekly patch-test reactions resulted in hyposensitization of the allergic contact dermatitis reaction. CD4CD25CD127lo/ regulatory T cells showed a tendency to increase; however, further studies are needed to determine if this is significant.
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Synthesis of new, UV-photoactive dansyl derivatives for flow cytometric studies on bile acid uptake.
Rohacova, Jana; Marin, M Luisa; Martínez-Romero, Alicia; O'Connor, José-Enrique; Gomez-Lechon, M Jose; Donato, M Teresa; Castell, Jose V; Miranda, Miguel A
2009-12-07
Four new fluorescent derivatives of cholic acid have been synthesized; they incorporate a dansyl moiety at 3alpha-, 3beta-, 7alpha- or 7beta- positions. These cholic acid analogs are UV photoactive and also exhibit green fluorescence. In addition, they have been demonstrated to be suitable for studying the kinetics of bile acid transport by flow cytometry.
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Eosin-sensitized photooxidation of substituted phenylalanines and tyrosines
Energy Technology Data Exchange (ETDEWEB)
Rizzuto, F.; Spikes, J.D.
1977-01-01
The cosin-sensitized photooxidation of tyrosine and a number of compounds related to tyrosine (substituted phenylalanines) was studied by steady-state kinetic and flash photolysis techniques. In particular, the role of the phenolic group and the amino and carboxyl groups of the alanyl side chain in the photooxidation mechanism was investigated in detail. Several relationships between substrate structure and susceptibility to photooxidation as well as effects of substrate structure on photooxidation mechanisms were found. For example, phenylalanine is not photooxidizable, but substitution of electron-donating (activating) groups such as -OH (as in tyrosine) or -NH/sub 2/ (as in p-aminophenylalanine) results in rapidly photooxidized derivatives. However, substituting deactivating groups such as -Cl (as in p-chlorophenylalanine) or weakly activating groups such as -OCH/sub 3/ (as in 4-methoxyphenylalanine) result in non-photooxidizable derivatives. Substitution of additional activating groups to the ring of hydroxy-substituted phenylalanines results in increased rates of photooxidation, whereas additional deactivating groups result in decreased photooxidation rates. The rate-determining step in the photooxidation mechanism is shown to be dependent on the presence and position of an electron-donating substituent on the benzenoid ring. Only minor involvement of the side chain amino and carboxyl groups was found. Both singlet oxygen and hydrogen abstraction mechanisms are involved in the eosin-sensitized photooxidation of hydroxy-substituted phenylalanines (e.g., tyrosine). The hydrogen abstraction mechanism probably predominates at both pH 8 and 11.
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Construction of BAC Libraries from Flow-Sorted Chromosomes.
Šafář, Jan; Šimková, Hana; Doležel, Jaroslav
2016-01-01
Cloned DNA libraries in bacterial artificial chromosome (BAC) are the most widely used form of large-insert DNA libraries. BAC libraries are typically represented by ordered clones derived from genomic DNA of a particular organism. In the case of large eukaryotic genomes, whole-genome libraries consist of a hundred thousand to a million clones, which make their handling and screening a daunting task. The labor and cost of working with whole-genome libraries can be greatly reduced by constructing a library derived from a smaller part of the genome. Here we describe construction of BAC libraries from mitotic chromosomes purified by flow cytometric sorting. Chromosome-specific BAC libraries facilitate positional gene cloning, physical mapping, and sequencing in complex plant genomes.
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Özyiğit, İbrahim Ethem; Karakuş, Emine; Pekcan, Önder
2016-02-01
Chymotrypsin and trypsin are the well known proteolytic enzymes, both of which are synthesized in the pancreas as their precursors - the inactive forms; chymotrypsinogen and trypsinogen - and then are released into the duodenum to cut proteins into smaller peptides. In this paper, the effects of activities of chymotrypsin and trypsin enzymes on fluorescence lifetime distributions of the substrat bovine serum albumin (BSA) modified with N-(1-pyrenyl)maleimide (PM) were examined. In the labeling study of BSA with PM, it is aimed to attach PM to the single free thiol (Cys34) and to all the free amine groups in accessible positions in order to produce excimers of pyrene planes of the possible highest amount to form the lifetime distributions in the widest range, that may show specifically distinguishing changes resulting from the activities of the proteases. The time resolved spectrofluorometer was used to monitor fluorescence decays, which were analyzed by using the exponential series method (ESM) to obtain the changes of lifetime distributions. After the exposure of the synthesized substrat PM-BSA to the enzymes, the fluorescence lifetime distributions exhibited different structures which were attributed to the different activities of the proteases.
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The photochemical conversion of solar energy into electrical energy: Eosin-Arabinose system
Energy Technology Data Exchange (ETDEWEB)
Gangotri, K.M. [Department of Chemistry, Solar Energy Laboratory, Jai Narain Vyas University, Jodhpur 342 033, Rajasthan (India); Bhimwal, Mukesh Kumar [Solar Energy Laboratory, Jai Narain Vyas University, Jodhpur 342 033, Rajasthan (India)
2010-12-15
A photosensitizer -Eosin and a reductant- Arabinose have been used in the photogalvanic cell for photochemical conversion of solar energy into electrical energy. The generated photopotential and photocurrent are 679.0 mV and 240.0 {mu}A respectively. The maximum power of the cell is 162.96 {mu}W whereas the observed power at power point is 73.08 {mu}W. The conversion efficiency is 0.7026% and the fill factor is 0.2856 at the power point of the photogalvanic cell. The photogalvanic cell so developed can work for 85.0 min in dark if it is irradiated for 140.0 min i.e. the storage capacity of photogalvanic cell is 60.71%. The effects of different parameters on the electrical output of the photogalvanic cell have been observed. A mechanism has also been proposed for the photogeneration of electrical energy. (author)
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Exfoliated graphite/titanium dioxide nanocomposites for photodegradation of eosin yellow
International Nuclear Information System (INIS)
Ndlovu, Thabile; Kuvarega, Alex T.; Arotiba, Omotayo A.; Sampath, Srinivasan; Krause, Rui W.; Mamba, Bhekie B.
2014-01-01
Graphical abstract: - Highlights: • Preparation of exfoliated graphite (EG) from natural graphite. • Sol–gel anchoring of TiO 2 on exfoliated graphite. • High adsorption and photoactivity was observed for the EG-TiO 2 nanocomposite. • Mechanism of enhancement was proposed. - Abstract: An improved photocatalyst consisting of a nanocomposite of exfoliated graphite and titanium dioxide (EG-TiO 2 ) was prepared. SEM and TEM micrographs showed that the spherical TiO 2 nanoparticles were evenly distributed on the surface of the EG sheets. A four times photocatalytic enhancement was observed for this floating nanocomposite compared to TiO 2 and EG alone for the degradation of eosin yellow. For all the materials, the reactions followed first order kinetics where for EG-TiO 2 , the rate constant was much higher than for EG and TiO 2 under visible light irradiation. The enhanced photocatalytic activity of EG-TiO 2 was ascribed to the capability of graphitic layers to accept and transport electrons from the excited TiO 2 , promoting charge separation. This indicates that carbon, a cheap and abundant material, can be a good candidate as an electron attracting reservoir for photocatalytic organic pollutant degradation
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Assessment of Flow Instability in Passive Auxiliary Feedwater System (PAFS) Using RELAP5
Energy Technology Data Exchange (ETDEWEB)
Jeon, Seong-Su; Hong, Soon-Joon [FNC Tech., Yongin (Korea, Republic of); Cheon, Jong; Kim, Han-Gon [KHNP, Daejeon (Korea, Republic of)
2015-10-15
In this study, the occurrence possibility of both instabilities in PAFS is assessed with the best-estimate thermal hydraulic code, RELAP5. From the RELAP5 code analysis, the Ledinegg instability might not occur in PAFS. The DWO might occur in PAFS but the effect of the oscillation on the heat removal capacity of PAFS was not large. Therefore, it is concluded that PAFS is safe in terms of flow instabilities. Since PAFS is two-phase flow system, flow instabilities may occur. Flow instabilities may cause the severe deterioration of heat removal capability of PAFS due to the reduction of the condensate flow. For the reliable operation of PAFS, it is required to assess the flow instabilities in PAFS. The Ledinegg-type instability and the Density Wave Oscillation (DWO) are the representative static flow instability and the dynamic flow instability, respectively.
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A study of the dispersed flow interfacial heat transfer model of RELAP5/MOD2.5 and RELAP5/MOD3
Energy Technology Data Exchange (ETDEWEB)
Andreani, M. [Swiss Federal Institute of Technology, Zurich (Switzerland); Analytis, G.T.; Aksan, S.N. [Paul Scherrer Institute, Villigen (Switzerland)
1995-09-01
The model of interfacial heat transfer for the dispersed flow regime used in the RELAP5 computer codes is investigated in the present paper. Short-transient calculations of two low flooding rate tube reflooding experiments have been performed, where the hydraulic conditions and the heat input to the vapour in the post-dryout region were controlled for the predetermined position of the quench front. Both RELAP5/MOD2.5 and RELAP5/MOD3 substantially underpredicted the exit vapour temperature. The mass flow rate and quality, however, were correct and the heat input to the vapour was larger than the actual one. As the vapour superheat at the tube exit depends on the balance between the heat input from the wall and the heat exchange with the droplets, the discrepancy between the calculated and the measured exit vapour temperature suggested that the inability of both codes to predict the vapour superheat in the dispersed flow region is due to the overprediction of the interfacial heat transfer rate.
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Use of internal control T-cell populations in the flow cytometric evaluation for T-cell neoplasms.
Hunt, Alicia M; Shallenberger, Wendy; Ten Eyck, Stephen P; Craig, Fiona E
2016-09-01
Flow cytometry is an important tool for identification of neoplastic T-cells, but immunophenotypic abnormalities are often subtle and must be distinguished from nonneoplastic subsets. Use of internal control (IC) T-cells in the evaluation for T-cell neoplasms was explored, both as a quality measure and as a reference for evaluating abnormal antigen expression. All peripheral blood specimens (3-month period), or those containing abnormal T-cells (29-month period), stained with CD45 V500, CD2 V450, CD3 PE-Cy7, CD7 PE, CD4 Per-CP-Cy5.5, CD8 APC-H7, CD56 APC, CD16&57 FITC, were evaluated. IC T-cells were identified (DIVA, BD Biosciences) and median fluorescence intensity (MFI) recorded. Selected files were merged and reference templates generated (Infinicyt, Cytognos). IC T-cells were present in all specimens, including those with abnormal T-cells, but subsets were less well-represented. IC T-cell CD3 MFI differed between instruments (p = 0.0007) and subsets (p < 0.001), but not specimen categories, and served as a longitudinal process control. Merged files highlighted small unusual IC-T subsets: CD2+(dim) (0.25% total), CD2- (0.03% total). An IC reference template highlighted neoplastic T-cells, but was limited by staining variability (IC CD3 MFI reference samples different from test (p = 0.003)). IC T-cells present in the majority of specimens can serve as positive and longitudinal process controls. Use of IC T-cells as an internal reference is limited by variable representation of subsets. Analysis of merged IC T-cells from previously analyzed patient samples can alert the interpreter to less-well-recognized non-neoplastic subsets. However, application of a merged file IC reference template was limited by staining variability. © 2016 Clinical Cytometry Society. © 2016 International Clinical Cytometry Society.
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Nieschke, Kathleen; Mittag, Anja; Golab, Karolina; Bocsi, Jozsef; Pierzchalski, Arkadiusz; Kamysz, Wojciech; Tarnok, Attila
2014-03-01
Toxicity test of new chemicals belongs to the first steps in the drug screening, using different cultured cell lines. However, primary human cells represent the human organism better than cultured tumor derived cell lines. We developed a very gentle toxicity assay for isolation and incubation of human peripheral blood leukocytes (PBL) and tested it using different bioactive oligopeptides (OP). Effects of different PBL isolation methods (red blood cell lysis; Histopaque isolation among others), different incubation tubes (e.g. FACS tubes), anticoagulants and blood sources on PBL viability were tested using propidium iodide-exclusion as viability measure (incubation time: 60 min, 36°C) and flow cytometry. Toxicity concentration and time-depended effects (10-60 min, 36 °C, 0-100 μg /ml of OP) on human PBL were analyzed. Erythrocyte lysis by hypotonic shock (dH2O) was the fastest PBL isolation method with highest viability (>85%) compared to NH4Cl-Lysis (49%). Density gradient centrifugation led to neutrophil granulocyte cell loss. Heparin anticoagulation resulted in higher viability than EDTA. Conical 1.5 mL and 2 mL micro-reaction tubes (both polypropylene (PP)) had the highest viability (99% and 97%) compared to other tubes, i.e. three types of 5.0 mL round-bottom tubes PP (opaque-60%), PP (blue-62%), Polystyrene (PS-64%). Viability of PBL did not differ between venous and capillary blood. A gentle reproducible preparation and analytical toxicity-assay for human PBL was developed and evaluated. Using our assay toxicity, time-course, dose-dependence and aggregate formation by OP could be clearly differentiated and quantified. This novel assay enables for rapid and cost effective multiparametric toxicological screening and pharmacological testing on primary human PBL and can be adapted to high-throughput-screening.°z
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Aydin, Z D; Barista, I; Canpinar, H; Sungur, A; Tekuzman, G
2000-07-01
In contrast to DNA ploidy, to the authors' knowledge the prognostic significance of S-phase fraction (SPF) in gastric lymphomas has not been determined. In the current study, the prognostic significance of various parameters including SPF and DNA aneuploidy were analyzed and some distinct epidemiologic and biologic features of gastric lymphomas in Turkey were found. A series of 78 gastric lymphoma patients followed at Hacettepe University is reported. DNA flow cytometry was performed for 34 patients. The influence of various parameters on survival was investigated with the log rank test. The Cox proportional hazards model was fitted to identify independent prognostic factors. The median age of the patients was 50 years. There was no correlation between patient age and tumor grade. DNA content analysis revealed 4 of the 34 cases to be aneuploid with DNA index values < 1.0. The mean SPF was 33.5%. In the univariate analysis, surgical resection of the tumor, modified Ann Arbor stage, performance status, response to first-line chemotherapy, lactate dehydrogenase (LDH) level, and SPF were important prognostic factors for disease free survival (DFS). The same parameters, excluding LDH level, were important for determining overall survival (OS). In the multivariate analysis, surgical resection of the tumor, disease stage, performance status, and age were found to be important prognostic factors for OS. To the authors' knowledge the current study is the first to demonstrate the prognostic significance of SPF in gastric lymphomas. The distinguishing features of Turkish gastric lymphoma patients are 1) DNA indices of aneuploid cases that all are < 1.0, which is a unique feature; 2) a lower percentage of aneuploid cases; 3) a higher SPF; 4) a younger age distribution; and 5) lack of an age-grade correlation. The authors conclude that gastric lymphomas in Turkey have distinct biologic and epidemiologic characteristics. Copyright 2000 American Cancer Society.
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PIC simulation of the vacuum power flow for a 5 terawatt, 5 MV, 1 MA pulsed power system
Liu, Laqun; Zou, Wenkang; Liu, Dagang; Guo, Fan; Wang, Huihui; Chen, Lin
2018-03-01
In this paper, a 5 Terawatt, 5 MV, 1 MA pulsed power system based on vacuum magnetic insulation is simulated by the particle-in-cell (PIC) simulation method. The system consists of 50 100-kV linear transformer drive (LTD) cavities in series, using magnetically insulated induction voltage adder (MIVA) technology for pulsed power addition and transmission. The pulsed power formation and the vacuum power flow are simulated when the system works in self-limited flow and load-limited flow. When the pulsed power system isn't connected to the load, the downstream magnetically insulated transmission line (MITL) works in the self-limited flow, the maximum of output current is 1.14 MA and the amplitude of voltage is 4.63 MV. The ratio of the electron current to the total current is 67.5%, when the output current reached the peak value. When the impedance of the load is 3.0 Ω, the downstream MITL works in the self-limited flow, the maximums of output current and the amplitude of voltage are 1.28 MA and 3.96 MV, and the ratio of the electron current to the total current is 11.7% when the output current reached the peak value. In addition, when the switches are triggered in synchronism with the passage of the pulse power flow, it effectively reduces the rise time of the pulse current.
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International Nuclear Information System (INIS)
Bloemen, Maarten; Verbiest, Thierry; Denis, Carla; Meester, Luc De; Peeters, Miet; Gils, Ann; Geukens, Nick
2015-01-01
We report on the design of superparamagnetic nanoparticles capable of selectively isolating targeted bacteria (Legionella pneumophila, serogroup 1) from aqueous solutions. The surface of magnetite nanoparticles (NP) was functionalized with a heterobifunctional poly(ethylene glycol) ligand containing reactive groups for covalent coupling of polyclonal antibodies against L. pneumophila. These bioconjugates were used to label and magnetically isolate L. pneumophila. Flow cytometry revealed high separation and efficiency in this regard. The strain specificity and efficiency of the magnetic NP was tested with recombinant strains of E. coli (expressing the red fluorescent protein) and L. pneumophila (expressing the green fluorescent protein). The detection limit of the method (by flow cytometry) is 10 4 cells∙mL -1 . The results indicate that the new multifunctional NPs are capable of selectively attracting pathogens from a complex mixture and with high efficiency. This, conceivably, paves the way to pre-concentration protocols for numerous other pathogens. (author)
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DEFF Research Database (Denmark)
Ring, Hans Christian; Riis, Peter Theut; Bay, Lene
2017-01-01
between bacterial aggregates identified by haematoxylin and eosin (HE) staining vs bacterial aggregates in corresponding PNA-FISH samples. Axillary biopsies were obtained in 24 healthy controls. HE-stained and PNA-FISH samples were investigated using traditional light microscopy and CLSM, respectively....... The data demonstrate that HE staining identifies large bacterial aggregates (>10 μm) with a sensitivity of 0.43 and specificity of 1. The methods, however, are not equivalent as demonstrated by a McNemar's test (P=.04). Where bacterial aggregates >10 μm in diameter, HE staining may offer a rapid...... and practical low-cost tool to evaluate bacterial aggregates....
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Relationship between aquaporin-5 expression and saliva flow in streptozotocin-induced diabetic mice?
Soyfoo, M S; Bolaky, N; Depoortere, I; Delporte, C
2012-07-01
To investigate the expression and distribution of AQP5 in submandibular acinar cells from sham- and streptozotocin (STZ)-treated mice in relation to the salivary flow. Mice were sham or STZ injected. Distribution of AQP5 subcellular expression in submandibular glands was determined by immunohistochemistry. AQP5 labelling indices (LI), reflecting AQP5 subcellular distribution, were determined in acinar cells. Western blotting was performed to determine the expression of AQP5 in submandibular glands. Blood glycaemia and osmolality and saliva flow rates were also determined. AQP5 immunoreactivity was primarily located at the apical and apical-basolateral membranes of submandibular gland acinar cells from sham- and STZ-treated mice. No significant differences in AQP5 protein levels were observed between sham- and STZ-treated mice. Compared to sham-treated mice, STZ-treated mice had significant increased glycaemia, while no significant differences in blood osmolality were observed. Saliva flow rate was significantly decreased in STZ-treated mice as compared to sham-treated mice. In STZ-treated mice, significant reduction in salivary flow rate was observed without any concomitant modification in AQP5 expression and localization. © 2011 John Wiley & Sons A/S.
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Sex-sorting sperm using flow cytometry/cell sorting.
Garner, Duane L; Evans, K Michael; Seidel, George E
2013-01-01
The sex of mammalian offspring can be predetermined by flow sorting relatively pure living populations of X- and Y-chromosome-bearing sperm. This method is based on precise staining of the DNA of sperm with the nucleic acid-specific fluorophore, Hoechst 33342, to differentiate between the subpopulations of X- and Y-sperm. The fluorescently stained sperm are then sex-sorted using a specialized high speed sorter, MoFlo(®) SX XDP, and collected into biologically supportive media prior to reconcentration and cryopreservation in numbers adequate for use with artificial insemination for some species or for in vitro fertilization. Sperm sorting can provide subpopulations of X- or Y-bearing bovine sperm at rates in the 8,000 sperm/s range while maintaining; a purity of 90% such that it has been applied to cattle on a commercial basis. The sex of offspring has been predetermined in a wide variety of mammalian species including cattle, swine, horses, sheep, goats, dogs, cats, deer, elk, dolphins, water buffalo as well as in humans using flow cytometric sorting of X- and Y-sperm.
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Prest, Emmanuelle I E C; Hammes, Frederik A.; Kö tzsch, Stefan; van Loosdrecht, Mark C.M.; Vrouwenvelder, Johannes S.
2013-01-01
Flow cytometry (FCM) is a rapid, cultivation-independent tool to assess and evaluate bacteriological quality and biological stability of water. Here we demonstrate that a stringent, reproducible staining protocol combined with fixed FCM operational and gating settings is essential for reliable quantification of bacteria and detection of changes in aquatic bacterial communities. Triplicate measurements of diverse water samples with this protocol typically showed relative standard deviation values and 95% confidence interval values below 2.5% on all the main FCM parameters. We propose a straightforward and instrument-independent method for the characterization of water samples based on the combination of bacterial cell concentration and fluorescence distribution. Analysis of the fluorescence distribution (or so-called fluorescence fingerprint) was accomplished firstly through a direct comparison of the raw FCM data and subsequently simplified by quantifying the percentage of large and brightly fluorescent high nucleic acid (HNA) content bacteria in each sample. Our approach enables fast differentiation of dissimilar bacterial communities (less than 15min from sampling to final result), and allows accurate detection of even small changes in aquatic environments (detection above 3% change). Demonstrative studies on (a) indigenous bacterial growth in water, (b) contamination of drinking water with wastewater, (c) household drinking water stagnation and (d) mixing of two drinking water types, univocally showed that this FCM approach enables detection and quantification of relevant bacterial water quality changes with high sensitivity. This approach has the potential to be used as a new tool for application in the drinking water field, e.g. for rapid screening of the microbial water quality and stability during water treatment and distribution in networks and premise plumbing. © 2013 Elsevier Ltd.
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Prest, Emmanuelle I E C
2013-12-01
Flow cytometry (FCM) is a rapid, cultivation-independent tool to assess and evaluate bacteriological quality and biological stability of water. Here we demonstrate that a stringent, reproducible staining protocol combined with fixed FCM operational and gating settings is essential for reliable quantification of bacteria and detection of changes in aquatic bacterial communities. Triplicate measurements of diverse water samples with this protocol typically showed relative standard deviation values and 95% confidence interval values below 2.5% on all the main FCM parameters. We propose a straightforward and instrument-independent method for the characterization of water samples based on the combination of bacterial cell concentration and fluorescence distribution. Analysis of the fluorescence distribution (or so-called fluorescence fingerprint) was accomplished firstly through a direct comparison of the raw FCM data and subsequently simplified by quantifying the percentage of large and brightly fluorescent high nucleic acid (HNA) content bacteria in each sample. Our approach enables fast differentiation of dissimilar bacterial communities (less than 15min from sampling to final result), and allows accurate detection of even small changes in aquatic environments (detection above 3% change). Demonstrative studies on (a) indigenous bacterial growth in water, (b) contamination of drinking water with wastewater, (c) household drinking water stagnation and (d) mixing of two drinking water types, univocally showed that this FCM approach enables detection and quantification of relevant bacterial water quality changes with high sensitivity. This approach has the potential to be used as a new tool for application in the drinking water field, e.g. for rapid screening of the microbial water quality and stability during water treatment and distribution in networks and premise plumbing. © 2013 Elsevier Ltd.
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Segovia, M.; Jenkins, J.A.; Paniagua-Chavez, C.; Tiersch, T.R.
2000-01-01
Improved techniques for storage and evaluation of fish sperm would enhance breeding programs around the world. The goal of this study was to test the effect of antibiotics on refrigerated sperm from Nile tilapia (Oreochromis niloticus) by use of flow cytometry with 2 dual-staining protocols for objective assessment of sperm quality. Concentrations of 1 x 109 sperm/mL were suspended in Ringer's buffer at 318 mOsmol/kg (pH 8.0). The fluorescent stains Sybr 14 (10 ??M), propidium iodide (2.4 mM), and rhodamine 123 (0.13 ??M) were used to assess cell viability and mitochondrial function. Three concentrations of ampicillin, gentamicin, and an antibiotic/antimycotic solution were added to fresh spermatozoa. Motility estimates and flow cytometry measurements were made daily during 7 d of refrigerated storage (4 ??C). The highest concentrations of gentamicin and antibiotic/antimycotic and all 3 concentrations of ampicillin significantly reduced sperm viability. The highest of each of the 3 antibiotic concentrations significantly reduced mitochondrial function. This study demonstrates that objective sperm quality assessments can be made using flow cytometry and that addition of antibiotics at appropriate concentrations can lengthen refrigerated storage time for tilapia spermatozoa. With minor modifications, these protocols can be adapted for use with sperm from other species and with other tissue types.
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Countercurrent flow limitation model for RELAP5/MOD3
International Nuclear Information System (INIS)
Riemke, R.A.
1991-01-01
This paper reports on a countercurrent flow limitation model incorporated into the RELAP5/MOD3 system transient analysis code. The model is implemented in a manner similar to the RELAP5 chocking model. Simulations using air/water flooding test problem demonstrate the ability of the code to significantly improve its comparison to data when a flooding correlation is used
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Detection of mycoplasmas in goat milk by flow cytometry.
Assunção, Patricia; Davey, Hazel M; Rosales, Ruben S; Antunes, Nuno T; de la Fe, Christian; Ramirez, Ana S; de Galarreta, Carlos M Ruiz; Poveda, Jose B
2007-12-01
The detection of mycoplasma in milk can be performed by either culture techniques or polymerase chain reaction (PCR) based methods. Although PCR can reduce the average diagnostic time to 5 h in comparison with the several days for the isolation of the agent, there is still a need to develop methods, which could give earlier results. For this purpose, we tested the ability of flow cytometry (FC) to detect mycoplasmas in milk samples. Milk samples inoculated with four different mycoplasmas, Mycoplasma agalactiae, Mycoplasma putrefaciens, Mycoplasma capricolum subsp. Capricolum, or Mycoplasma mycoides subsp. mycoides large-colony type, known to cause contagious agalactia in goats, were stained with the DNA stain SYBR Green I and analyzed by FC. Three goat milk samples, from which mycoplasmas have been isolated in broth medium were also analyzed. All mycoplasmas were easily distinguished from debris of milk samples, but it was not possible to distinguish between the different mycoplasma species. In our conditions, the detection limit of the technique was of the order of 10(3)-10(4) cells ml(-1). Furthermore, mycoplasmas were also distinguished from Staphylococcus aureus. FC together with SYBR Green I was able to distinguish between mycoplasma cells and debris present in milk samples and gave results in 20-30 min. This is an important first step in developing a robust, routine flow cytometric method for the detection of mycoplasmas in milk samples. (c) 2007 International Society for Analytical Cytology
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Exfoliated graphite/titanium dioxide nanocomposites for photodegradation of eosin yellow
Energy Technology Data Exchange (ETDEWEB)
Ndlovu, Thabile, E-mail: atkuvarega@gmail.com [University of Swaziland, Department of Chemistry, Private Bag 4, Kwaluseni (Swaziland); Kuvarega, Alex T.; Arotiba, Omotayo A. [University of Johannesburg, Department of Applied Chemistry, P.O. Box 17011, Doornfontein 2028, Johannesburg (South Africa); Sampath, Srinivasan [Indian Institute of Science, Department of Inorganic and Physical Chemistry, Bangalore 560012 (India); Krause, Rui W. [Rhodes University, Department of Chemistry, P.O. Box 94, Grahamstown 6140 South Africa (South Africa); Mamba, Bhekie B., E-mail: bmamba@uj.ac.za [University of Johannesburg, Department of Applied Chemistry, P.O. Box 17011, Doornfontein 2028, Johannesburg (South Africa)
2014-05-01
Graphical abstract: - Highlights: • Preparation of exfoliated graphite (EG) from natural graphite. • Sol–gel anchoring of TiO{sub 2} on exfoliated graphite. • High adsorption and photoactivity was observed for the EG-TiO{sub 2} nanocomposite. • Mechanism of enhancement was proposed. - Abstract: An improved photocatalyst consisting of a nanocomposite of exfoliated graphite and titanium dioxide (EG-TiO{sub 2}) was prepared. SEM and TEM micrographs showed that the spherical TiO{sub 2} nanoparticles were evenly distributed on the surface of the EG sheets. A four times photocatalytic enhancement was observed for this floating nanocomposite compared to TiO{sub 2} and EG alone for the degradation of eosin yellow. For all the materials, the reactions followed first order kinetics where for EG-TiO{sub 2}, the rate constant was much higher than for EG and TiO{sub 2} under visible light irradiation. The enhanced photocatalytic activity of EG-TiO{sub 2} was ascribed to the capability of graphitic layers to accept and transport electrons from the excited TiO{sub 2}, promoting charge separation. This indicates that carbon, a cheap and abundant material, can be a good candidate as an electron attracting reservoir for photocatalytic organic pollutant degradation.
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Özyiğit, İbrahim Ethem; Karakuş, Emine; Pekcan, Önder
2016-02-05
Chymotrypsin and trypsin are the well known proteolytic enzymes, both of which are synthesized in the pancreas as their precursors - the inactive forms; chymotrypsinogen and trypsinogen - and then are released into the duodenum to cut proteins into smaller peptides. In this paper, the effects of activities of chymotrypsin and trypsin enzymes on fluorescence lifetime distributions of the substrat bovine serum albumin (BSA) modified with N-(1-pyrenyl)maleimide (PM) were examined. In the labeling study of BSA with PM, it is aimed to attach PM to the single free thiol (Cys34) and to all the free amine groups in accessible positions in order to produce excimers of pyrene planes of the possible highest amount to form the lifetime distributions in the widest range, that may show specifically distinguishing changes resulting from the activities of the proteases. The time resolved spectrofluorometer was used to monitor fluorescence decays, which were analyzed by using the exponential series method (ESM) to obtain the changes of lifetime distributions. After the exposure of the synthesized substrat PM-BSA to the enzymes, the fluorescence lifetime distributions exhibited different structures which were attributed to the different activities of the proteases. Copyright © 2015 Elsevier B.V. All rights reserved.
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FLOW CYTOMETRIC APPLICABILITY OF FLUORESCENT VITALITY PROBES ON PHYTOPLANKTON1.
Peperzak, Louis; Brussaard, Corina P D
2011-06-01
The applicability of six fluorescent probes (four esterase probes: acetoxymethyl ester of Calcein [Calcein-AM], 5-chloromethylfluorescein diacetate [CMFDA], fluorescein diacetate [FDA], and 2',7'-dichlorofluorescein diacetate [H 2 DCFDA]; and two membrane probes: bis-(1,3-dibutylbarbituric acid) trimethine oxonol [DiBAC 4 (3)] and SYTOX-Green) as vitality stains was tested on live and killed cells of 40 phytoplankton strains in exponential and stationary growth phases, belonging to 12 classes and consisting of four cold-water, 26 temperate, and four warm-water species. The combined live/dead ratios of all six probes indicated significant differences between the 12 plankton classes (P live/dead ratios of FDA and CMFDA were not significantly different from each other, and both performed better than Calcein-AM and H 2 DCFDA (P live/dead ratios) among all six probes belonged to nine genera from six classes of phytoplankton. In conclusion, FDA, CMFDA, DIBAC 4 (3), and SYTOX-Green represent a wide choice of vitality probes in the study of phytoplankton ecology, applicable in many species from different algal classes, originating from different regions and at different stages of growth. © 2011 Phycological Society of America.
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Angra 2 small break LOCA flow regime identification through RELAP5 code
Energy Technology Data Exchange (ETDEWEB)
Rocha, Marcelo da Silva; Sabundjian, Gaiane; Belchior Junior, Antonio; Andrade, Delvonei Alves de; Torres, Walmir Maximo; Conti, Thadeu das Neves; Macedo, Luiz Alberto; Umbehaun, Pedro Ernesto; Mesquita, Roberto Navarro de; Masotti, Paulo Henrique Ferraz, E-mail: msrocha@ipen.br, E-mail: gdjian@ipen.br, E-mail: abelchior@ipen.br, E-mail: delvonei@ipen.br, E-mail: wmtorres@ipen.br, E-mail: tnconti@ipen.br, E-mail: lamacedo@ipen.br, E-mail: umbehaun@ipen.br, E-mail: s, E-mail: rnavarro@ipen.br, E-mail: pmasotti@ipen.br [Instituto de Pesquisas Energeticas e Nucleares (IPEN/CNEN-RJ), Rio de Janeiro, RJ (Brazil)
2012-07-01
The purpose of this paper is to identify the flow regimes in the core of Angra 2 nuclear reactor with RELAP5/MOD3.2.gamma code (RELAP5, 2001). The postulated accident is the loss of coolant through a small break in the primary circuit (SBLOCA), which is described in Chapter 15 of the Final Safety Analysis Report of Angra 2 - FSAR (ETN, 2006). As the primary circuit pressure decreases due to the loss of coolant, several alternating two phase flow regimes are established in the primary circuit. This paper analyses the coolant two-phase flow behavior in the nuclear reactor core during the postulated accident. (author)
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Biology and flow cytometry of proangiogenic hematopoietic progenitors cells.
Rose, Jonathan A; Erzurum, Serpil; Asosingh, Kewal
2015-01-01
During development, hematopoiesis and neovascularization are closely linked to each other via a common bipotent stem cell called the hemangioblast that gives rise to both hematopoietic cells and endothelial cells. In postnatal life, this functional connection between the vasculature and hematopoiesis is maintained by a subset of hematopoietic progenitor cells endowed with the capacity to differentiate into potent proangiogenic cells. These proangiogenic hematopoietic progenitors comprise a specific subset of bone marrow (BM)-derived cells that homes to sites of neovascularization and possess potent paracrine angiogenic activity. There is emerging evidence that this subpopulation of hematopoietic progenitors plays a critical role in vascular health and disease. Their angiogenic activity is distinct from putative "endothelial progenitor cells" that become structural cells of the endothelium by differentiation into endothelial cells. Proangiogenic hematopoietic progenitor cell research requires multidisciplinary expertise in flow cytometry, hematology, and vascular biology. This review provides a comprehensive overview of proangiogenic hematopoietic progenitor cell biology and flow cytometric methods to detect these cells in the peripheral blood circulation and BM. © 2014 International Society for Advancement of Cytometry.
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Determination of Complement-Mediated Killing of Bacteria by Viability Staining and Bioluminescence
Virta, Marko; Lineri, Sanna; Kankaanpää, Pasi; Karp, Matti; Peltonen, Karita; Nuutila, Jari; Lilius, Esa-Matti
1998-01-01
Complement-mediated killing of bacteria was monitored by flow cytometric, luminometric, and conventional plate counting methods. A flow cytometric determination of bacterial viability was carried out by using dual staining with a LIVE/DEAD BacLight bacterial viability kit. In addition to the viable cell population, several other populations emerged in the fluorescence histogram, and there was a dramatic decrease in the total cell count in the light-scattering histogram in the course of the co...
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Study of the Eosin-Y/PAMAM interactions in alkaline aqueous solution
Energy Technology Data Exchange (ETDEWEB)
Arbeloa, Ernesto M., E-mail: earbeloa@exa.unrc.edu.ar [Universidad Nacional de Río Cuarto, Río Cuarto, 5800 Córdoba (Argentina); Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET) (Argentina); Previtali, Carlos M. [Universidad Nacional de Río Cuarto, Río Cuarto, 5800 Córdoba (Argentina); Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET) (Argentina); Bertolotti, Sonia G., E-mail: sbertolotti@exa.unrc.edu.ar [Universidad Nacional de Río Cuarto, Río Cuarto, 5800 Córdoba (Argentina); Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET) (Argentina)
2016-04-15
The interactions between the xanthene dye Eosin-Y (Eos) and amino-terminated PAMAM dendrimers of low generations (G0–G3) were studied in alkaline water solution. The effect of concentration and generation of the dendrimer on the photophysics of Eos was evaluated by means of absorption and fluorescence spectroscopies. The observed spectral changes were ascribed to the association dye/dendrimer. From these data, the Eos/PAMAM binding constants (K{sub bind}) were determined, which strongly increased with the size of the dendrimer. Stationary fluorescence anisotropy and time-resolved single photon counting were also used to characterize the association process. The restriction in the rotational diffusion of the Eos increased as a function of the concentration and generation of PAMAM, as determined by anisotropy measurements. Biexponential fluorescence decays were obtained in the presence of G3, and the respective lifetimes were ascribed to free and bound Eos species. These results correlate with K{sub bind} values and suggest the formation of host/guest system with larger dendrimers. Therefore, this environmentally-friendly dye/dendrimer system would be appropriate for potential applications in fields such as drugs delivery and photopolymerization.
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Study of the Eosin-Y/PAMAM interactions in alkaline aqueous solution
International Nuclear Information System (INIS)
Arbeloa, Ernesto M.; Previtali, Carlos M.; Bertolotti, Sonia G.
2016-01-01
The interactions between the xanthene dye Eosin-Y (Eos) and amino-terminated PAMAM dendrimers of low generations (G0–G3) were studied in alkaline water solution. The effect of concentration and generation of the dendrimer on the photophysics of Eos was evaluated by means of absorption and fluorescence spectroscopies. The observed spectral changes were ascribed to the association dye/dendrimer. From these data, the Eos/PAMAM binding constants (K bind ) were determined, which strongly increased with the size of the dendrimer. Stationary fluorescence anisotropy and time-resolved single photon counting were also used to characterize the association process. The restriction in the rotational diffusion of the Eos increased as a function of the concentration and generation of PAMAM, as determined by anisotropy measurements. Biexponential fluorescence decays were obtained in the presence of G3, and the respective lifetimes were ascribed to free and bound Eos species. These results correlate with K bind values and suggest the formation of host/guest system with larger dendrimers. Therefore, this environmentally-friendly dye/dendrimer system would be appropriate for potential applications in fields such as drugs delivery and photopolymerization.
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Improved and Reproducible Flow Cytometry Methodology for Nuclei Isolation from Single Root Meristem
Directory of Open Access Journals (Sweden)
Thaís Cristina Ribeiro Silva
2010-01-01
Full Text Available Root meristems have increasingly been target of cell cycle studies by flow cytometric DNA content quantification. Moreover, roots can be an alternative source of nuclear suspension when leaves become unfeasible and for chromosome analysis and sorting. In the present paper, a protocol for intact nuclei isolation from a single root meristem was developed. This proceeding was based on excision of the meristematic region using a prototypical slide, followed by short enzymatic digestion and mechanical isolation of nuclei during homogenization with a hand mixer. Such parameters were optimized for reaching better results. Satisfactory nuclei amounts were extracted and analyzed by flow cytometry, producing histograms with reduced background noise and CVs between 3.2 and 4.1%. This improved and reproducible technique was shown to be rapid, inexpensive, and simple for nuclear extraction from a single root tip, and can be adapted for other plants and purposes.
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Directory of Open Access Journals (Sweden)
Fangjun Yao
2017-03-01
Full Text Available The impacts of hydrogen bonding on polymerization behavior has been of interest for a long time; however, universality and in-depth understanding are still lacking. For the first time, the effect of hydrogen bonding on the classical alternating-type copolymerization of styrene and maleimide was explored. N-phenylmaleimide (N-PMI/styrene was chosen as a model monomer pair in the presence of hydrogen bonding donor solvent 1,1,1,3,3,3-hexafluoro-2-propanol (HFIP, which interacted with N-PMI via hydrogen bonding. Reversible addition-fragmentation chain transfer polymerization (RAFT technique was used to guarantee the “living” polymerization and thus the homogeneity of chain compositions. In comparison with the polymerization in nonhydrogen bonding donor solvent (toluene, the copolymerization in HFIP exhibited a high rate and a slight deviation from alternating copolymerization tendency. The reactivity ratios of N-PMI and St were revealed to be 0.078 and 0.068, respectively, while the reactivity ratios in toluene were 0.026 and 0.050. These interesting results were reasonably explained by using computer simulations, wherein the steric repulsion and electron induction by the hydrogen bonding between HFIP and NPMI were revealed. This work first elucidated the hydrogen bonding interaction in the classical alternating-type copolymerization, which will enrich the research on hydrogen bonding-induced polymerizations.
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Fungal Fluorescence in Hematoxylin-Eosin Stained Sections
Directory of Open Access Journals (Sweden)
Murat Durdu
2017-06-01
Full Text Available A forty-six-year-old male presented to our dermatology clinic with two-year history of itching on his groin. His medical history revealed various topical corticosteroid creams without improvement of the skin lesion. Dermatological examination revealed erythematous nodules and follicular pustules on erythematous background on the inguinal area (Figure 1a. Potassium hydroxide (KOH examination was negative. Tzanck smear revealed abundant neutrophils without bacteria, fungi, or parasite. The histopathological examination showed granuloma formation with multinuclear giant cells and Periodic acid-Schiff (PAS-positive hyphae and spores around the hair follicles (Figure 1b, 1c. Hematoxylin-eosin (H&E-stained slides were examined under an immunofluorescence microscope, and these hyphae and spores showed autofluorescence (Figure 1d. Based on the clinical and histopathological findings, a Majocchi’s granuloma was considered. All lesions disappeared with topical and systemic terbinafine (250 mg/day treatment for six weeks.\tPearls;\tClinical: Not only bacteria, but also fungi, parasites, and viruses may cause folliculitis. Cytology should be initially done to identify the causes of infectious folliculitis. In case of negative cytology, histopathological examination and molecular methods can be used.\tCytological: To cytologically identify all of the causes of folliculitis, four separate samples should be taken: (i the first sample is stained with May-Grünwald-Giemsa for routine cytological examination; (ii the second sample is used for KOH testing; (iii the third sample is stained with an acid-fast stain to detect mycobacteria; and (iv the last specimen is Gram-stained to identify whether it is Gram-positive or Gram-negative (1. Histopathological: In infectious diseases, a definitive diagnosis should be done to identify the etiologic agent. The detection of fungal elements is challenging, when histopathological examination is performed with the H
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Flow-cytometric determination of high-density-lipoprotein binding sites on human leukocytes
International Nuclear Information System (INIS)
Schmitz, G.; Wulf, G.; Bruening, T.A.; Assmann, G.
1987-01-01
In this method, leukocytes were isolated from 6 mL of EDTA-blood by density-gradient centrifugation and subsequently incubated with rhodamine isothiocyanate (RITC)-conjugated high-density lipoproteins (HDL). The receptor-bound conjugate particles were determined by fluorescent flow cytometry and compared with 125 I-labeled HDL binding data for the same cells. Human granulocytes express the highest number of HDL binding sites (9.4 x 10(4)/cell), followed by monocytes (7.3 x 10(4)/cell) and lymphocytes (4.0 x 10(4)/cell). Compared with conventional analysis of binding of 125 I-labeled HDL in tissue-culture dishes, the present determination revealed significantly lower values for nonspecific binding. In competition studies, the conjugate competes for the same binding sites as 125 I-labeled HDL. With the use of tetranitromethane-treated HDL3, which fails to compete for the HDL receptor sites while nonspecific binding is not affected, we could clearly distinguish between 37 degrees C surface binding and specific 37 degrees C uptake of RITC-HDL3, confirming that the HDL receptor leads bound HDL particles into an intracellular pathway rather than acting as a docking type of receptor. Patients with familial dysbetalipoproteinemia showed a significantly higher number of HDL binding sites in the granulocyte population but normal in lymphocytes and monocytes, indicating increased uptake of cholesterol-containing lipoproteins. In patients with familial hypercholesterolemia, HDL binding was increased in all three cell types, indicating increased cholesterol uptake and increased cholesterol synthesis. The present method allows rapid determination of HDL binding sites in leukocytes from patients with various forms of hyper- and dyslipoproteinemias
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Bunthof, C.J.; Abee, T.
2002-01-01
Flow cytometry (FCM) is a rapid and sensitive technique that can determine cell numbers and measure various physiological characteristics of individual cells by using appropriate fluorescent probes. Previously, we developed an FCM assay with the viability probes carboxyfluorescein diacetate (cFDA)
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Luo, Teng; Lu, Yuan; Liu, Shaoxiong; Lin, Danying; Qu, Junle
2017-09-05
The phasor approach to fluorescence lifetime imaging microscopy (FLIM) is used to identify different types of tissues from hematoxylin and eosin (H&E) stained basal cell carcinoma (BCC) sections. The results suggest that working directly on the phasor space with the clustering assignment achieves immunofluorescence like simultaneous five or six-color imaging by using multiplexed fluorescence lifetimes of H&E. The phase approach is of particular effectiveness for enhanced visualization of the abnormal morphology of a suspected nidus. Moreover, the phasor approach to H&E FLIM data can determine the actual paths or the infiltrating trajectories of basophils and immune cells associated with the preneoplastic or neoplastic skin lesions. The integration of the phasor approach with routine histology proved its available value for skin cancer prevention and early detection. We therefore believe that the phasor analysis of H&E tissue sections is an enhanced visualization tool with the potential to simplify the preparation process of special staining and serve as color contrast aided imaging in clinical pathological examination.
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Spectrophotometric determination of neodymium in mixture with lanthanum by eosin and 2,2'-dipyridyl
International Nuclear Information System (INIS)
Ovchar, L.A.; Poluehktov, N.S.
1980-01-01
The possibility of using rare earth complexes with eosin (EO) and 2.2-dipyridyl (DP) for spectrophotometric determination of some rare earths in the presence of the others. It has been out that the complexes are not extracted by organic solvents. The PH region of complex existence (approximately 6) and the relation of components in them (rare earths:DP:EO=1:2:3) are determined. The possibility has been shown of determining all the rare earts from praseodymium to lutetium and yttrium in a binary mixture with lanthanum based on different stability of the studied complexes. The method has been tested on the example of determining Nd 2 O 3 in a mixture with La 2 O 3 . The low limit of determined contents is 1-2%. The relative standard deviation is 0.035-0.17 [ru
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Energy Technology Data Exchange (ETDEWEB)
Manickasundaram, S. [Department of Chemistry, Anna University, Sardar Vallabai Patel Road, Chennai 600025 (India); Kannan, P. [Department of Chemistry, Anna University, Sardar Vallabai Patel Road, Chennai 600025 (India)]. E-mail: pakannan@annauniv.edu; Deepa, S. [Centre for Laser Technology, Department of Physics, Anna University, Chennai 600025 (India); Palanisamy, P.K. [Centre for Laser Technology, Department of Physics, Anna University, Chennai 600025 (India)
2007-01-15
A new series of eosin dye based poly(alkyloxymethacrylate)s was synthesized with an even number of side-chain methylene spacers by a free radical addition polymerization method for holographic optical data storage applications. These polymers were characterized by UV, IR and {sup 1}H NMR spectroscopy. The glass transition temperature and thermal stability of the polymers were investigated by DSC and TGA, respectively. As the spacer length increases in the side-chain, Tg, Tm and thermal stability of the polymers decrease, while a reverse trend was observed with film forming ability of the polymers. The optical characterization of the polymers was investigated by forming holographic grating using an Argon ion laser. The grating diffraction efficiency was found to depend not only on the concentration of polymeric film but also on the spacer length of the polymers.
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International Nuclear Information System (INIS)
Manickasundaram, S.; Kannan, P.; Deepa, S.; Palanisamy, P.K.
2007-01-01
A new series of eosin dye based poly(alkyloxymethacrylate)s was synthesized with an even number of side-chain methylene spacers by a free radical addition polymerization method for holographic optical data storage applications. These polymers were characterized by UV, IR and 1 H NMR spectroscopy. The glass transition temperature and thermal stability of the polymers were investigated by DSC and TGA, respectively. As the spacer length increases in the side-chain, Tg, Tm and thermal stability of the polymers decrease, while a reverse trend was observed with film forming ability of the polymers. The optical characterization of the polymers was investigated by forming holographic grating using an Argon ion laser. The grating diffraction efficiency was found to depend not only on the concentration of polymeric film but also on the spacer length of the polymers
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Automated Blood Sample Preparation Unit (ABSPU) for Portable Microfluidic Flow Cytometry.
Chaturvedi, Akhil; Gorthi, Sai Siva
2017-02-01
Portable microfluidic diagnostic devices, including flow cytometers, are being developed for point-of-care settings, especially in conjunction with inexpensive imaging devices such as mobile phone cameras. However, two pervasive drawbacks of these have been the lack of automated sample preparation processes and cells settling out of sample suspensions, leading to inaccurate results. We report an automated blood sample preparation unit (ABSPU) to prevent blood samples from settling in a reservoir during loading of samples in flow cytometers. This apparatus automates the preanalytical steps of dilution and staining of blood cells prior to microfluidic loading. It employs an assembly with a miniature vibration motor to drive turbulence in a sample reservoir. To validate performance of this system, we present experimental evidence demonstrating prevention of blood cell settling, cell integrity, and staining of cells prior to flow cytometric analysis. This setup is further integrated with a microfluidic imaging flow cytometer to investigate cell count variability. With no need for prior sample preparation, a drop of whole blood can be directly introduced to the setup without premixing with buffers manually. Our results show that integration of this assembly with microfluidic analysis provides a competent automation tool for low-cost point-of-care blood-based diagnostics.
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Analysis of UV-excited fluorochromes by flow cytometry using near-ultraviolet laser diodes.
Telford, William G
2004-09-01
Violet laser diodes have become common and reliable laser sources for benchtop flow cytometers. While these lasers are very useful for a variety of violet and some ultraviolet-excited fluorochromes (e.g., DAPI), they do not efficiently excite most UV-stimulated probes. In this study, the next generation of InGaN near-UV laser diodes (NUVLDs) emitting in the 370-375-nm range have been evaluated as laser sources for cuvette-based flow cytometers. Several NUVLDs, ranging in wavelength from 370 to 374 nm and in power level from 1.5 to 10 mW, were mounted on a BD Biosciences LSR II and evaluated for their ability to excite cells labeled with the UV DNA binding dye DAPI, several UV phenotyping fluorochromes (including Alexa Fluor 350, Marina Blue, and quantum dots), and the fluorescent calcium chelator indo-1. NUVLDs at the 8-10-mW power range gave detection sensitivity levels comparable to more powerful solid-state and ion laser sources, using low-fluorescence microsphere beads as measurement standards. NUVLDs at all tested power levels allowed extremely high-resolution DAPI cell cycle analysis, and sources in the 8-10-mW power range excited Alexa Fluor 350, Marina Blue, and a variety of quantum dots at virtually the same signal-to-noise ratios as more powerful UV sources. These evaluations indicate that near-UV laser diodes installed on a cuvette-based flow cytometer performed nearly as well as more powerful solid-state UV lasers on the same instrumentation, and comparably to more powerful ion lasers on a jet-in-air system, and. Despite their limited power, integration of these small and inexpensive lasers into benchtop flow cytometers should allow the use of flow cytometric applications requiring UV excitation on a wide variety of instruments. Copyright 2004 Wiley-Liss, Inc.
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Directory of Open Access Journals (Sweden)
Yutaro Shiota
2000-01-01
Full Text Available Study objectives were to compare the numbers of lymphocytes and lymphocyte subpopulations in induced sputum from asthmatic patients and from healthy subjects, and to determine the effect of inhaled anti-asthmatic steroid therapy on these cell numbers. Hypertonic saline inhalation was used to non-invasively induce sputum samples in 34 patients with bronchial asthma and 21 healthy subjects. The sputum samples were reduced with dithioerythritol and absolute numbers of lymphocytes and lymphocyte subpopulations were assessed by direct immunofluorescence and flow cytometry. To assess the effect of beclomethasone dipropionate (BDP on induced sputum, numbers of lymphocytes and lymphocyte subpopulations in sputum also were evaluated after 4 weeks of BDP inhalation treatment in seven asthmatic patients. An adequate sample was obtained in 85.3% of patients with asthma and in 79.2% of the healthy subjects. Induced sputum from patients with asthma had increased numbers of lymphocytes (P = 0.009; CD4+ cells (P = 0.044; CD4+ cells-bearing interleukin-2 receptor (CD25; P = 0.016; and CD4+ cells bearing human histocompatibility leukocyte antigen (HLA-DR (P = 0.033. CD8+ cells were not increased in asthmatic patients. In patients treated with inhaled steroids, numbers of lymphocytes, CD4+ cells, CD25-bearing CD4+ cells and HLA-DR-bearing CD4+ cells in sputum decreased from pretreatment numbers (P = 0.016, 0.002, 0.003 and 0.002, respectively. Analysis of lymphocytes in induced sputum by flow cytometry is useful in assessing bronchial inflammation, and activated CD4+ lymphocytes may play a key role in the pathogenesis of airway inflammation in bronchial asthma.
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International Nuclear Information System (INIS)
Cao Wei; Li Jing; Wu Zhiqun; Zhou Changxi; Liu Xi; Wan Yi; Duan Yunyou
2013-01-01
Purpose. This study evaluates the influence of transcatheter arterial infusion with heated saline on hepatic arterial and portal venous blood flows to tumor and normal hepatic tissues in a rabbit VX2 tumor model. Methods. All animal experiments were approved by the institutional animal care and use committee. Twenty rabbits with VX2 liver tumors were divided into the following two groups: (a) the treated group (n = 10), which received a 60 mL transarterial injection of 60 °C saline via the hepatic artery; (b) the control group (n = 10), which received a 60 mL injection of 37 °C saline via the hepatic artery. Using ultrasonography, the blood flows in both the portal vein and hepatic artery were measured, and the changes in the hemodynamic indices were recorded before and immediately after the injection. The changes in the tumor and normal liver tissues of the two groups were histopathologically examined by hematoxylin and eosin staining after the injection. Results. After the transcatheter arterial heated infusion, there was a decrease in the hepatic arterial blood flow to the tumor tissue, a significant decrease in the hepatic artery mean velocity (P < 0.05), and a significant increase in the resistance index (P < 0.05). On hematoxylin and eosin staining, there were no obvious signs of tissue destruction in the normal liver tissue or the tumor tissue after heated perfusion, and coagulated blood plasma was observed in the cavities of intratumoral blood vessels in the treated group. Conclusions. The changes in tumor blood flow in the rabbit VX2 tumor model were presumably caused by microthrombi in the tumor vessels, and the portal vein likely mediated the heat loss in normal liver tissue during the transarterial heated infusion.
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Energy Technology Data Exchange (ETDEWEB)
Cao Wei, E-mail: cawe-001@163.com [Tangdu Hospital, The Fourth Military Medical University, Department of Interventional Radiology (China); Li Jing, E-mail: lijing02@fmmu.edu.cn [Tangdu Hospital, The Fourth Military Medical University, Department of Burn and Plastic Surgery (China); Wu Zhiqun, E-mail: zhiqunwu@fmmu.edu.cn [Tangdu Hospital, The Fourth Military Medical University, Department of Interventional Radiology (China); Zhou Changxi, E-mail: changxizhou@163.com [Chinese PLA General Hospital, Department of Respiratory Disease (China); Liu Xi, E-mail: xiliu@fmmu.edu.cn [Tangdu Hospital, The Fourth Military Medical University, Department of Ultrasound Diagnostics (China); Wan Yi, E-mail: yiwan@163.com [The Fourth Military Medical University, Department of Health Statistics, Institute for Health Informatics (China); Duan Yunyou, E-mail: yunyouduan@fmmu.edu.cn [Tangdu Hospital, The Fourth Military Medical University, Department of Ultrasound Diagnostics (China)
2013-06-15
Purpose. This study evaluates the influence of transcatheter arterial infusion with heated saline on hepatic arterial and portal venous blood flows to tumor and normal hepatic tissues in a rabbit VX2 tumor model. Methods. All animal experiments were approved by the institutional animal care and use committee. Twenty rabbits with VX2 liver tumors were divided into the following two groups: (a) the treated group (n = 10), which received a 60 mL transarterial injection of 60 Degree-Sign C saline via the hepatic artery; (b) the control group (n = 10), which received a 60 mL injection of 37 Degree-Sign C saline via the hepatic artery. Using ultrasonography, the blood flows in both the portal vein and hepatic artery were measured, and the changes in the hemodynamic indices were recorded before and immediately after the injection. The changes in the tumor and normal liver tissues of the two groups were histopathologically examined by hematoxylin and eosin staining after the injection. Results. After the transcatheter arterial heated infusion, there was a decrease in the hepatic arterial blood flow to the tumor tissue, a significant decrease in the hepatic artery mean velocity (P < 0.05), and a significant increase in the resistance index (P < 0.05). On hematoxylin and eosin staining, there were no obvious signs of tissue destruction in the normal liver tissue or the tumor tissue after heated perfusion, and coagulated blood plasma was observed in the cavities of intratumoral blood vessels in the treated group. Conclusions. The changes in tumor blood flow in the rabbit VX2 tumor model were presumably caused by microthrombi in the tumor vessels, and the portal vein likely mediated the heat loss in normal liver tissue during the transarterial heated infusion.
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Reticulocyte maturity index by flow cytometry: its applicability in radioinduced bone marrow aplasia
International Nuclear Information System (INIS)
Dubner, D.; Gisone, P.; Perez, M.R.
1995-01-01
Flow cytometric reticulocyte quantification was assayed in ten patients undergoing bone marrow transplantation (BMT) with previous conditioning chemotherapy and total body irradiation (TBI). A reticulocyte maturity index (RMI) was determined taking into account the RNA content. With de aim of testing the utility of RMI as an early predictor of functional recovery in marrow aplasia, other haematological indicators as neutrophils count were comparatively evaluated. Mean time elapsed between BMT and engraftment evidence by RMI was 17,6 days. In six patients the RMI was the earliest indicator of functional recovery. The applicability of this assay in the following of radioinduced bone marrow aplasia is discussed. (author). 4 refs., 4 figs., 2 tabs
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Props, Ruben; Schmidt, Marian L; Heyse, Jasmine; Vanderploeg, Henry A; Boon, Nico; Denef, Vincent J
2018-02-01
Species invasion is an important disturbance to ecosystems worldwide, yet knowledge about the impacts of invasive species on bacterial communities remains sparse. Using a novel approach, we simultaneously detected phenotypic and derived taxonomic change in a natural bacterioplankton community when subjected to feeding pressure by quagga mussels, a widespread aquatic invasive species. We detected a significant decrease in diversity within 1 h of feeding and a total diversity loss of 11.6 ± 4.1% after 3 h. This loss of microbial diversity was caused by the selective removal of high nucleic acid populations (29 ± 5% after 3 h). We were able to track the community diversity at high temporal resolution by calculating phenotypic diversity estimates from flow cytometry (FCM) data of minute amounts of sample. Through parallel FCM and 16S rRNA gene amplicon sequencing analysis of environments spanning a broad diversity range, we showed that the two approaches resulted in highly correlated diversity measures and captured the same seasonal and lake-specific patterns in community composition. Based on our results, we predict that selective feeding by invasive dreissenid mussels directly impacts the microbial component of the carbon cycle, as it may drive bacterioplankton communities toward less diverse and potentially less productive states. © 2017 Society for Applied Microbiology and John Wiley & Sons Ltd.
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Directory of Open Access Journals (Sweden)
Josep M. Gasol
2000-06-01
Full Text Available Flow cytometry is rapidly becoming a routine methodology in aquatic microbial ecology. The combination of simple to use bench-top flow cytometers and highly fluorescent nucleic acid stains allows fast and easy determination of microbe abundance in the plankton of lakes and oceans. The different dyes and protocols used to stain and count planktonic bacteria as well as the equipment in use are reviewed, with special attention to some of the problems encountered in daily routine practice such as fixation, staining and absolute counting. One of the main advantages of flow cytometry over epifluorescence microscopy is the ability to obtain cell-specific measurements in large numbers of cells with limited effort. We discuss how this characteristic has been used for differentiating photosynthetic from non-photosynthetic prokaryotes, for measuring bacterial cell size and nucleic acid content, and for estimating the relative activity and physiological state of each cell. We also describe how some of the flow cytometrically obtained data can be used to characterize the role of microbes on carbon cycling in the aquatic environment and we prospect the likely avenues of progress in the study of planktonic prokaryotes through the use of flow cytometry.
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Kargahi, Neda; Razavi, Sayyed Mohammad; Deyhimi, Parviz; Homayouni, Solmaz
2015-01-01
Oral squamous cell carcinoma is the most common malignant lesion of the oral cavity, and it involves various molecular mechanisms. The development of oral squamous cell carcinoma is influenced by the host immune cells, such as eosinophils. The present study was conducted to compare the presence of eosinophils in normal mucosa, dysplastic mucosa, and oral squamous cell carcinoma by -hematoxylin- eosin staining, Congo red staining, and epidermal growth factor-like (EGF-like) module containing a mucin-like hormone receptor1 (EMR1) immunohistochemical marker. In this cross-sectional study, 60 paraffinized samples were selected, consisting of 20 normal mucosae, 20 dysplastic mucosae, and 20 squamous cell carcinoma samples. After confirmation of the diagnosis, the mean number of eosinophils was evaluated by hematoxylin-eosin, Congo red, and immunohystochemical staining techniques. The data were analyzed by SPSS-10 software using the Kruskal-Wallis and Friedman tests. The results showed that the number of eosinophils in dysplastic mucosa was significantly higher than the number in normal mucosa, and the number of eosinophils in squamous cell carcinoma was significantly higher than the number in dysplastic mucosa in all staining techniques (p<0.001). Moreover, the comparison of staining techniques showed a significantly higher number of eosinophils in EMR1immunohistochemicalmarker than were observed when Congo red and hematoxylin - eosin (H&E) staining techniques were used (p<0.001). It can be argued that eosinophil contributes to the identification of lesions that have a higher potential of malignant transformation. Moreover, eosinophil can be suggested as an indicator in the differentiation of oral lesions in cases with borderline diagnosis and in targeted molecular therapy.
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Konokhova, Anastasiya I; Chernova, Darya N; Moskalensky, Alexander E; Strokotov, Dmitry I; Yurkin, Maxim A; Chernyshev, Andrei V; Maltsev, Valeri P
2016-02-01
Importance of microparticles (MPs), also regarded as extracellular vesicles, in many physiological processes and clinical conditions motivates one to use the most informative and precise methods for their characterization. Methods based on individual particle analysis provide statistically reliable distributions of MP population over characteristics. Although flow cytometry is one of the most powerful technologies of this type, the standard forward-versus-side-scattering plots of MPs and platelets (PLTs) overlap considerably because of similarity of their morphological characteristics. Moreover, ordinary flow cytometry is not capable of measurement of size and refractive index (RI) of MPs. In this study, we 1) employed the potential of the scanning flow cytometer (SFC) for identification and characterization of MPs from light scattering; 2) suggested the reference method to characterize MP morphology (size and RI) with high precision; and 3) determined the lowest size of a MP that can be characterized from light scattering with the SFC. We equipped the SFC with 405 and 488 nm lasers to measure the light-scattering profiles and side scattering from MPs, respectively. The developed two-stage method allowed accurate separation of PLTs and MPs in platelet-rich plasma. We used two optical models for MPs, a sphere and a bisphere, in the solution of the inverse light-scattering problem. This solution provides unprecedented precision in determination of size and RI of individual spherical MPs-median uncertainties (standard deviations) were 6 nm and 0.003, respectively. The developed method provides instrument-independent quantitative information on MPs, which can be used in studies of various factors affecting MP population. © 2015 International Society for Advancement of Cytometry.
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Laminar and Turbulent Flow Calculations for the Hifire-5B Flight Test
2017-11-01
STATES AIR FORCE AFRL-RQ-WP-TP-2017-0172 LAMINAR AND TURBULENT FLOW CALCULATIONS FOR THE HIFIRE-5B FLIGHT TEST Roger L. Kimmel Hypersonic Sciences...stationary instabilities of the three-dimensional flow as the grid becomes finer. It may not be possible to obtain a strictly laminar basic state on a very...fine grid. A basic state solution was desired for the laminar flow calculations, and the oscillations observed in Fig. 3 were judged to be undesirable
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Directory of Open Access Journals (Sweden)
Malik Abu Rafee
2017-12-01
Full Text Available The study was conducted on four healthy guinea pigs (Cavia porcellus of either sex in which the nerve was identified and subjected to crush injury with the tip (3mm of a curved hemostatic forceps. 30 days after the injury nerve samples were collected and subjected to Hematoxylin and Eosin staining with or without pretreatment with Marchi’s solution. The routine Hematoxylin and Eosin (H&E stained all neural elements in various intensities of pink and in purple and the degenerative changes were seen as vacuoles ranging from vacuolated foci- containing eosinophilic material and associated with a distorted cell nucleus to larger, multilocular, linear array of compartmentalized digestion chambers supposed to contain myelin debris .The myelin on the other hand appeared as empty zones in H&E staining. Combining Marchi’s and H & E procedures revealed the presence black aggregates/ deposits in the vacuoles and digestion chambers. This method confirmed the presence of degenerated myelin inside the vacuoles and digestion chambers and thus may allow better analysis of nerve damage and regeneration.
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Directory of Open Access Journals (Sweden)
Alanna Mara Pinheiro Sobreira Bezerra
2011-06-01
Full Text Available Objective: To demonstrate the advantages of correlatingflow cytometry immunophenotyping with the pathology/immunohistochemistry of lymph nodes or nodules in the diagnosisof lymphoproliferative diseases. Methods: A retrospective studywas carried out of 157 biopsy or fine-needle aspiration lymph nodes/nodule specimens taken from 142 patients, from 1999 and 2009.The specimens were simultaneously studied with flow cytometryand pathology at Hospital Israelita Albert Einstein. The specimenswere prepared in hematoxylin/eosin, Giemsa, or monoclonal antibodystained slides for detecting specific antibodies for the purposesof pathology/immunohistochemical analysis. The samples werehemolyzed and marked with different monoclonal antibody panels fordifferent antigens in flow cytometry immunophenotyping. Results:The diagnostic results of pathology/immunohistochemical studiesand flow cytometry immunophenotyping agreed in 115 patients(81%, corresponding to 127 specimens, as follows according tothe pathologic diagnosis: 63 patients with non-Hodgkin’s B-celllymphoma; 26 patients with reactive lymphoid hyperplasia; 5 patientswith non-Hodgkin’s T-cell lymphoma; 4 patients with atypical lymphoidproliferation; 5 patients with a chronic granulomatous inflammatoryprocess; 5 patients with a non-hematologic diagnosis; 2 patientswith granulocytic sarcoma; 2 patients with thymoma; 1 patientwith byphenotypic leukemia; 1 patient with kappa plasmocytoma;1 patient with Hodgkin’s lymphoma. Subtypes of lymphomas couldbe classified by associating the two techniques: 19 patients withfollicular lymphoma; 15 patients with diffuse large B-cell lymphoma; 7patients with small lymphocytic B-cell lymphoma/chronic lymphocyticleukemia; 3 patients with mantle cell lymphoma; 1 patient withBurkitt’s lymphoma; 1 patient with MALT type lymphoma; 1 patientwith post-transplant lymphoproliferative disease; 2 patients with highgrade non-Hodgkin’s B-cell lymphoma; 1 patient with low grade
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Directory of Open Access Journals (Sweden)
T. S. Anirudhan
2015-01-01
Full Text Available The present study describes the feasibility of a novel adsorbent cum photocatalyst, poly(pyrrole-co-aniline-coated TiO2/nanocellulose composite (P(Py-co-An-TiO2/NCC, to remove eosin yellow (EY from aqueous solutions. The removal of EY was investigated by batch adsorption followed by photocatalysis. The effect of various adsorption parameters like adsorbent dose, pH, contact time, initial concentration, and ionic strength has been optimized for treating effluents from the dye industry. Adsorption of EY reached maximum at pH 4.5 and complete removal of dye was achieved using 3.5 g/L of P(Py-co-An-TiO2/NCC. Adsorption equilibrium data were fitted with Langmuir and Fritz-Schlunder isotherm models and the kinetics of adsorption follows a second-order mechanism. The adsorption capacity of P(Py-co-An-TiO2/NCC was found to be 3.39 × 10−5 mol/g and reached equilibrium within 90 min. The photocatalytic degradation of adsorbed dye under sunlight was possible and about 92.3% of dye was degraded within 90 min. The reusability of P(Py-co-An-TiO2/NCC was also investigated. The results indicate that P(Py-co-An-TiO2/NCC is the best material for the wiping out of EY from aqueous solutions.
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Tessmann, Josiane Weber; Buss, Julieti; Begnini, Karine Rech; Berneira, Lucas Moraes; Paula, Favero Reisdorfer; de Pereira, Claudio Martin Pereira; Collares, Tiago; Seixas, Fabiana Kömmling
2017-10-01
Bladder cancer is a genitourinary malignant disease common worldwide. Current chemotherapy is often limited mainly due to toxicity and drug resistance. Thus, there is a continued need to discover new therapies. Recently evidences shows that pyrazoline derivatives are promising antitumor agents in many types of cancers, but there are no studies with bladder cancer. In order to find potent and novel chemotherapy drugs for bladder cancer, a series of pyrazoline derivatives 2a-2d were tested for their antitumor activity in two human bladder cancer cell lines 5647 and T24. The MTT assay showed that the compounds 1-thiocarbamoyl-3,5-diphenyl-4,5-dihydro-1H-pyrazole (2a) and 1-thiocarbamoyl-5-(4-chlorophenyl)-3-phenyl-4,5-dihydro-1H-pyrazole (2c) decrease the cell viability of 5637 cells. Molecular modeling indicated that these compounds had a good oral bioavailability and low toxicities. Clonogenic assay and flow cytometric analysis were used to assess colony formation, apoptosis induction and cell cycle distribution. Overall, our results suggest that pyrazoline 2a and 2c, with the substituents hydrogen and chlorine respectively, may decrease cell viability and colony formation of bladder cancer 5637 cell line by inhibition of cell cycle progression, and for pyrazoline 2a, by induction of apoptosis. As indicated by the physicochemical properties of these compounds, the steric factor influences the activity. Therefore, these pyrazoline derivatives can be considered promising anticancer agents for the treatment of bladder cancer. Copyright © 2017 Elsevier Masson SAS. All rights reserved.
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A Flow Cytometry Protocol to Estimate DNA Content in the Yellowtail Tetra Astyanax altiparanae
Directory of Open Access Journals (Sweden)
Pedro L. P. Xavier
2017-09-01
Full Text Available The production of triploid yellowtail tetra Astyanax altiparanae is a key factor to obtain permanently sterile individuals by chromosome set manipulation. Flow cytometric analysis is the main tool for confirmation of the resultant triploids individuals, but very few protocols are specific for A. altiparanae species. The current study has developed a protocol to estimate DNA content in this species. Furthermore, a protocol for long-term storage of dorsal fins used for flow cytometry analysis was established. The combination of five solutions with three detergents (Nonidet P-40 Substitute, Tween 20, and Triton X-100 at 0.1, 0.2, and 0.4% concentration was evaluated. Using the best solution from this first experiment, the addition of trypsin (0.125, 0.25, and 0.5% and sucrose (74 mM and the effects of increased concentrations of the detergents at 0.6 and 1.2% concentration were also evaluated. After adjustment of the protocol for flow cytometry, preservation of somatic tissue or isolated nuclei was also evaluated by freezing (at −20°C and fixation in saturated NaCl solution, acetic methanol (1:3, ethanol, and formalin at 10% for 30 or 60 days of storage at 25°C. Flow cytometry analysis in yellowtail tetra species was optimized using the following conditions: lysis solution: 9.53 mM MgCl2.7H20; 47.67 mM KCl; 15 mM Tris; 74 mM sucrose, 0.6% Triton X-100, pH 8.0; staining solution: Dulbecco's PBS with DAPI 1 μg mL−1; preservation procedure: somatic cells (dorsal fin samples frozen at −20°C. Using this protocol, samples may be stored up to 60 days with good accuracy for flow cytometry analysis.
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Dual fluorescence and laser emissions from fluorescein-Na and eosin-B
International Nuclear Information System (INIS)
Math, N.N.; Naik, L.R.; Suresh, H.M.; Inamdar, S.R.
2006-01-01
Dual laser emissions were observed from fluorescein-Na and eosin-B in ethanolic solutions individually in the concentration range from 10 -2 to 10 -3 mol dm -3 under N 2 laser excitation. The first compound was found to lase at two distinct regions with wavelength maxima around 540, 550 nm, while the second one around 558, 574 nm. Steady-state absorption, fluorescence excitation, fluorescence polarization, fluorescence emission and decays of the dyes in various solvents under varying conditions of excitation and detection systems were carried out to identify the nature of the emitting species responsible for laser emissions in two distinct regions. Both the dyes exhibited concentration and excitation wavelength dependence of fluorescence and the effects were found to be more pronounced in binary solution. The fluorescence decays of dyes were monoexponential in ethanol, while in some other solvents used, the decays showed biexponential behavior. The absorption and excitation studies using thin layers of solutions revealed the formation of dimers with the dye concentration around 1x10 -3 mol dm -3 . Fluorescence polarization and decay studies confirmed the presence of dimers. The two laser bands observed in the shorter and longer wavelengths were respectively ascribed to monomeric and dimeric species
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Dual fluorescence and laser emissions from fluorescein-Na and eosin-B
Energy Technology Data Exchange (ETDEWEB)
Math, N.N. [Laser Spectroscopy (DRDO/KU) Programme, Department of Physics, Karnatak University, Dharwad 580 003 (India)]. E-mail: nnm31@rediffmail.com; Naik, L.R. [Laser Spectroscopy (DRDO/KU) Programme, Department of Physics, Karnatak University, Dharwad 580 003 (India); Suresh, H.M. [Laser Spectroscopy (DRDO/KU) Programme, Department of Physics, Karnatak University, Dharwad 580 003 (India); Inamdar, S.R. [Laser Spectroscopy (DRDO/KU) Programme, Department of Physics, Karnatak University, Dharwad 580 003 (India)
2006-12-15
Dual laser emissions were observed from fluorescein-Na and eosin-B in ethanolic solutions individually in the concentration range from 10{sup -2} to 10{sup -3} mol dm{sup -3} under N{sub 2} laser excitation. The first compound was found to lase at two distinct regions with wavelength maxima around 540, 550 nm, while the second one around 558, 574 nm. Steady-state absorption, fluorescence excitation, fluorescence polarization, fluorescence emission and decays of the dyes in various solvents under varying conditions of excitation and detection systems were carried out to identify the nature of the emitting species responsible for laser emissions in two distinct regions. Both the dyes exhibited concentration and excitation wavelength dependence of fluorescence and the effects were found to be more pronounced in binary solution. The fluorescence decays of dyes were monoexponential in ethanol, while in some other solvents used, the decays showed biexponential behavior. The absorption and excitation studies using thin layers of solutions revealed the formation of dimers with the dye concentration around 1x10{sup -3} mol dm{sup -3}. Fluorescence polarization and decay studies confirmed the presence of dimers. The two laser bands observed in the shorter and longer wavelengths were respectively ascribed to monomeric and dimeric species.
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Directory of Open Access Journals (Sweden)
Michael S Bono
Full Text Available In this study, we cultured Chlorella vulgaris cells with a range of lipid contents, induced via nitrogen starvation, and characterized them via flow cytometry, with BODIPY 505/515 as a fluorescent lipid label, and liquid-state 1H NMR spectroscopy. In doing so, we demonstrate the utility of calibrating flow cytometric measurements of algal lipid content using triacylglyceride (TAG, also known as triacylglycerol or triglyceride content per cell as measured via quantitative 1H NMR. Ensemble-averaged fluorescence of BODIPY-labeled cells was highly correlated with average TAG content per cell measured by bulk NMR, with a linear regression yielding a linear fit with r2 = 0.9974. This correlation compares favorably to previous calibrations of flow cytometry protocols to lipid content measured via extraction, and calibration by NMR avoids the time and complexity that is generally required for lipid quantitation via extraction. Flow cytometry calibrated to a direct measurement of TAG content can be used to investigate the distribution of lipid contents for cells within a culture. Our flow cytometry measurements showed that Chlorella vulgaris cells subjected to nitrogen limitation exhibited higher mean lipid content but a wider distribution of lipid content that overlapped the relatively narrow distribution of lipid content for replete cells, suggesting that nitrogen limitation induces lipid accumulation in only a subset of cells. Calibration of flow cytometry protocols using direct in situ measurement of TAG content via NMR will facilitate rapid development of more precise flow cytometry protocols, enabling investigation of algal lipid accumulation for development of more productive algal biofuel feedstocks and cultivation protocols.
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Bono Jr., Michael S.; Garcia, Ravi D.; Sri-Jayantha, Dylan V.; Ahner, Beth A.; Kirby, Brian J.
2015-01-01
In this study, we cultured Chlorella vulgaris cells with a range of lipid contents, induced via nitrogen starvation, and characterized them via flow cytometry, with BODIPY 505/515 as a fluorescent lipid label, and liquid-state 1H NMR spectroscopy. In doing so, we demonstrate the utility of calibrating flow cytometric measurements of algal lipid content using triacylglyceride (TAG, also known as triacylglycerol or triglyceride) content per cell as measured via quantitative 1H NMR. Ensemble-averaged fluorescence of BODIPY-labeled cells was highly correlated with average TAG content per cell measured by bulk NMR, with a linear regression yielding a linear fit with r 2 = 0.9974. This correlation compares favorably to previous calibrations of flow cytometry protocols to lipid content measured via extraction, and calibration by NMR avoids the time and complexity that is generally required for lipid quantitation via extraction. Flow cytometry calibrated to a direct measurement of TAG content can be used to investigate the distribution of lipid contents for cells within a culture. Our flow cytometry measurements showed that Chlorella vulgaris cells subjected to nitrogen limitation exhibited higher mean lipid content but a wider distribution of lipid content that overlapped the relatively narrow distribution of lipid content for replete cells, suggesting that nitrogen limitation induces lipid accumulation in only a subset of cells. Calibration of flow cytometry protocols using direct in situ measurement of TAG content via NMR will facilitate rapid development of more precise flow cytometry protocols, enabling investigation of algal lipid accumulation for development of more productive algal biofuel feedstocks and cultivation protocols. PMID:26267664
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Bono, Michael S; Garcia, Ravi D; Sri-Jayantha, Dylan V; Ahner, Beth A; Kirby, Brian J
2015-01-01
In this study, we cultured Chlorella vulgaris cells with a range of lipid contents, induced via nitrogen starvation, and characterized them via flow cytometry, with BODIPY 505/515 as a fluorescent lipid label, and liquid-state 1H NMR spectroscopy. In doing so, we demonstrate the utility of calibrating flow cytometric measurements of algal lipid content using triacylglyceride (TAG, also known as triacylglycerol or triglyceride) content per cell as measured via quantitative 1H NMR. Ensemble-averaged fluorescence of BODIPY-labeled cells was highly correlated with average TAG content per cell measured by bulk NMR, with a linear regression yielding a linear fit with r2 = 0.9974. This correlation compares favorably to previous calibrations of flow cytometry protocols to lipid content measured via extraction, and calibration by NMR avoids the time and complexity that is generally required for lipid quantitation via extraction. Flow cytometry calibrated to a direct measurement of TAG content can be used to investigate the distribution of lipid contents for cells within a culture. Our flow cytometry measurements showed that Chlorella vulgaris cells subjected to nitrogen limitation exhibited higher mean lipid content but a wider distribution of lipid content that overlapped the relatively narrow distribution of lipid content for replete cells, suggesting that nitrogen limitation induces lipid accumulation in only a subset of cells. Calibration of flow cytometry protocols using direct in situ measurement of TAG content via NMR will facilitate rapid development of more precise flow cytometry protocols, enabling investigation of algal lipid accumulation for development of more productive algal biofuel feedstocks and cultivation protocols.
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International Nuclear Information System (INIS)
Fletcher, M.P.; Seligmann, B.E.
1985-01-01
Purified human peripheral blood polymorphonuclear neutrophils (PMN) were incubated at 37 degrees C with the fluorescent membrane potential sensitive cyanine dye di-O-C(5)(3) and exposed to a number of stimulatory agents (N-formylmethionylleucylphenylalanine (FMLP), cytochalasin B (cyto B) + FMLP, phorbol myristate acetate (PMA). Flow cytometry was utilized to measure changes in forward light scatter (FS), orthogonal light scatter (90 degrees-SC), and fluorescence intensity of individual cells over time. A saturating (10(-6) M) dose of FMLP lead to a significant increase in the cells' FS without a change in 90 degrees-SC as well as a heterogeneous loss of di-O-C(5)(3) fluorescence. PMA (100 ng/ml) also caused an increase in FS but a uniform loss of dye fluorescence by all cells (apparent depolarization). Cyto B + FMLP produced an increase in FS, a marked loss of 90 degrees-SC, and a uniform loss of fluorescence. Secretion experiments under identical incubation conditions indicated a significantly positive relationship between loss of enzyme markers or cell granularity and orthogonal light scatter (r . 0.959, 0.998, and 0.989 for loss of 90 degrees-SC vs lysozyme, beta-glucuronidase, and granularity index, respectively). Flow cytometric light scatter measurements may yield important information on the extent of prior cell degranulation or activation
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faloabi@uniben.edu Antiproliferative and Pro-apoptotic activities
African Journals Online (AJOL)
MICHAEL HORSFALL
Keyword: Persea americana, antiproliferative activity, apoptotic effect, flow ... of the stem bark of Persea americana in MCF-7 cell line by flow cytometer. .... of an electric milling machine. ... Flow Cytometric Measurement Of Cell Proliferation:.
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Contribution of flow cytometry to the diagnosis of gastric lymphomas in endoscopic biopsy specimens.
Almasri, N M; Zaer, F S; Iturraspe, J A; Braylan, R C
1997-07-01
Gastric lymphomas seem to have unique clinical, pathologic, and immunophenotypic features that set them apart from nodal lymphomas. Microscopic examination of endoscopic biopsy specimens is the most frequent procedure used to diagnose gastric tumors, but it is very difficult, and sometimes impossible, to recognize lymphomas in endoscopic samples by histologic or even immunohistologic methods. Because most gastric lymphomas are of B-cell origin, we used flow cytometry to assess B-cell clonality in gastric biopsy specimens containing dense lymphocytic infiltrates thought to represent lymphoma. We prepared viable cell suspensions from unfixed specimens obtained from 29 consecutive patients who had a previous microscopic diagnosis of suspicious gastric lymphoid infiltrates. We performed immunophenotypic studies with multicolor flow cytometry, and we assessed clonality by examination of immunoglobulin (Ig) light-chain expression analyzed exclusively on B cells identified by anti-CD20 or CD19 antibodies. The mean number of cells recovered was 1.04 x 10(6), from an average of 5.5 gastric biopsy fragments per patient. In 26 of the 29 patients, the number of cells was adequate for analysis. We detected B-cell monoclonality in 16 cases, including 5 in which the percentage of clonal B cells was less than 5%. Of the 16 cases, only 8 could be diagnosed as lymphomas on morphologic grounds alone; the remaining 8 patients had either suspicious lymphoid infiltrates or chronic gastritis. The three cases with an insufficient number of cells were considered non-neoplastic either on histologic grounds alone or in conjunction with Southern analysis of Ig genes. We conclude that flow cytometric immunophenotypic analysis of freshly prepared cell suspensions obtained from endoscopic biopsy specimens can be used to evaluate gastric lymphocytic infiltrates. Specifically, the analysis of surface Ig light-chain expression on B cells distinguishes between monoclonal (lymphoma) and polyclonal
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Location of radiation-induced grafted chains in polymers studied by solid-state NMR
International Nuclear Information System (INIS)
Whittacker, A.; Liu, H.
1998-01-01
In this study styrene and N-phenyl maleimide monomers were grafted onto poly(ethylene) (PE) chains using gamma radiation. Of main interest is the distribution of grafted chains within the polymer matrix, as this will determine the efficacy of mixing with the glassy polymers. It is expected that grafting will occur within the amorphous regions, and especially near the interface of the crystalline and amorphous regions. A suitable method for characterising the location of the grafted chains is solid-state 13 C NMR spectroscopy. The 13 C CPMAS spectrum of the blend of PE and N-phenyl maleimide mixed in the melt at 150 deg C , prior to reaction, is shown above. The spectrum shows the typical peaks for poly(ethylene) due to the amorphous and crystalline phase at 30.5 and 32.5 ppm, respectively. Peaks are also seen in the aromatic and carbonyl region due to the maleimide (not plotted). Experiments will be described where the NMR magnetisation is prepared in either the crystalline and amorphous regions of the poly(ethylene) prior to spin diffusion to the maleimide and styrene fractions. The location of the grafted monomers can then be determined by monitoring the changes in signal of polymer and graft with time
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Directory of Open Access Journals (Sweden)
Kenji Nagata
2015-09-01
Full Text Available Purpose: To report a case of CD5+ primary intraocular B-cell lymphoma arising during methotrexate (MTX and tumor necrosis factor (TNF inhibitor treatment in a young patient with rheumatoid arthritis and uveitis. Case Presentation: A 39-year-old woman treated with MTX and a TNF inhibitor for rheumatoid arthritis and uveitis had steroid-resistant vitreous opacity. A vitreous sample was obtained by using diagnostic vitrectomy and was categorized as class V based on cytologic examination. Flow cytometric analysis of the vitreous sample revealed that abnormal cells were CD5+, CD10-, CD19+, CD20+ and immunoglobulin light-chain kappa+, suggesting the diagnosis of CD5+ primary intraocular B-cell lymphoma. Polymerase chain reaction (PCR detected immunoglobulin heavy-chain gene rearrangement. Epstein-Barr virus (EBV DNA was detected in the vitreous sample by using PCR, and immunohistochemistry revealed EBV latent membrane protein-1 expression in the abnormal cells infiltrating the vitreous. Optic nerve invasion was observed on magnetic resonance imaging. Conclusion: Primary intraocular lymphoma (PIOL may develop in patients receiving MTX and TNF inhibitor treatment. EBV infection may play an important role in the pathogenesis of PIOL arising during immunosuppressive therapy.
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High-definition hematoxylin and eosin staining in a transition to digital pathology
Directory of Open Access Journals (Sweden)
Jamie D Martina
2011-01-01
Full Text Available Introduction: A lot of attention has been generated in recent years by digital pathology and telepathology. Multiple reasons for and barriers to effective adoption are discussed in the current literature. Digital slides are the most promising medium at this time. The goal of our study was to evaluate whether the change in the methodology, particularly utilizing the so-called high-definition hematoxylin and eosin (H and E slides, enhanced the quality of the final digital slide, and whether pathologists who tested the results perceived this as a difference in quality. Methods: The study was a blinded comparison of digital slides prepared using two methods: standard H&E batch staining and automated individual "high definition" HD HE staining. Four pathologists have compared 80 cases stained with each method. Results: The results discussed in this study show potential promise that the utilization of protocol(s adapted for tissue and for imaging might be preferable for digital pathology in at least some of the pathology subspecialties. In particular, the protocol evaluated here was capable of turning out digital slides that had more contrast and detail, and therefore were perceived to provide enhanced diagnostically significant information for the pathologist.
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Energy Technology Data Exchange (ETDEWEB)
Puangpetch, Tarawipa [Department of Chemical Engineering, Faculty of Engineering and Industrial Technology, Silpakorn University, Nakhon Pathom 73000 (Thailand); Sommakettarin, Pichayaon [The Petroleum and Petrochemical College, Chulalongkorn University, Soi Chula 12, Phyathai Road, Pathumwan, Bangkok 10330 (Thailand); Chavadej, Sumaeth; Sreethawong, Thammanoon [The Petroleum and Petrochemical College, Chulalongkorn University, Soi Chula 12, Phyathai Road, Pathumwan, Bangkok 10330 (Thailand); Center for Petroleum, Petrochemicals, and Advanced Materials, Chulalongkorn University, Bangkok 10330 (Thailand)
2010-11-15
The photocatalytic water splitting is a promising process for producing H{sub 2} from two abundant renewable sources of water and solar light, with the aid of a suitable photocatalyst. In this work, a combination of sensitizer addition and noble metal loading was employed to modify perovskite photocatalysts in order to achieve the enhancement of photocatalytic H{sub 2} production under visible light irradiation. The dependence of the H{sub 2} production on type of mesoporous-assembled perovskite titanate nanocrystal photocatalysts (MgTiO{sub 3}, CaTiO{sub 3}, and SrTiO{sub 3}), calcination temperature of photocatalyst, Pt loading, type and concentration of electron donor (diethanolamine, DEA; and triethanolamine, TEA), concentration of sensitizer (Eosin Y, E.Y.), photocatalyst dosage, and initial solution pH, was systematically studied. The experimental results showed that the 0.5 wt.% Pt-loaded mesoporous-assembled SrTiO{sub 3} nanocrystal synthesized by a single-step sol-gel method and calcined at 650 C exhibited the highest photocatalytic H{sub 2} production activity from a 15 vol% DEA aqueous solution with dissolved 0.5 mM E.Y. Moreover, the optimum photocatalyst dosage and initial solution pH for the maximum photocatalytic H{sub 2} production activity were found to be 6 g/l and 11.6, respectively. (author)
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Li, Dayu; Liu, Yun; Yu, Chunbo; Liu, Xiping; Fan, Fang
2017-12-01
Objective To study the effect of the knock-down of the DNA-dependent protein kinase catalytic subunit (DNA-PKcs) on the cell cycle of the multidrug-resistant (MDR) Bel7402/5-Fu hepatocellular carcinoma cells and its MDR mechanism. Methods After cationic liposome-mediated siDNA-PKcs oligonucleotide transfection, the drug sensitivity of Bel7402/5-Fu cells to 5-fluorouracil (5-Fu) and adriamycin (ADM) was determined by MTT assay; the cell cycle were detected by flow cytometry; meanwhile, the protein expressions of cell cycle-related proteins P21, cell cycle protein B1 (cyclin B1), cell cycle division protein 2 (CDC2) were tested by Western blotting; the expressions of ataxia telangiectasia mutated (ATM) and p53 at both mRNA and protein levels were detected by real-time PCR and Western blot analysis. Results The MTT results showed siDNA-PKcs increased the chemotherapeutic sensitivity of Bel7402/5-Fu cells to 5-Fu and ADM. The flow cytometric analysis showed siDNA-PKcs decreased the percentage of S-phase cells but increased the percentage of G2/M phase cells. Western blotting showed siDNA-PKcs increased the protein expression of P21 but decreased cyclinB1 and CDC2 proteins. In addition, siDNA-PKcs also increased the expressions of ATM and p53. Conclusion DNA-PKcs silencing increases P21 while decreases cyclin B1 and CDC2 expressions, and finally induces G2/M phase arrest in Bel7402/5-Fu cells, which may be related to ATM-p53 signaling pathway.
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Zhang, Ning; Shi, Jinwen; Niu, Fujun; Wang, Jian; Guo, Liejin
2015-09-07
Owing to the effect of energy band bending, p-type Co3O4 quantum dots sensitized by Eosin Y showed a high and stable photocatalytic activity (∼13,440 μmol h(-1) g(-1)(cat)) for water reduction and hydrogen production under visible-light irradiation without any cocatalyst.
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Eosin yellowish dye sensitized TiO2 solar cell with PEG/PEO/LiI/I2 as electrolyte
Kanmani, S. S.; Umapathy, S.; Ramachandran, K.
2012-06-01
Eosin Yellowish dye sensitized TiO2 nanoparticles (NP) and nanowires (NW) are employed as photo anodes in dye sensitized solar cells with PEO/PEG/LiI/I2 as electrolyte. Material characterization by XRD and SEM confirms the formation of anatase phased TiO2 NP and NW. Effective quenching of UV emission in TiO2 NW than NP is a consequence of reduction in recombination rate, which directly favours for better solar conversion efficiency. The photovoltaic performance of TiO2 NW with an overall conversion efficiency of 0.31 % is better than NP, which is the outcome of improved electron transport in NW.
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G.J.A. Arkesteijn (Ger); C.A.J. Erpelinck (Claudia); A.C.M. Martens (Anton); A. Hagenbeek (Anton)
1995-01-01
textabstractFlow cytometry was used to measure the fluorescence intensity of nuclei that were subjected to fluorescent in situ hybridization in suspension with chromosome specific DNA probes. Paraformaldehyde-fixed nuclei were protein digested with trypsin and hybridized simultaneously with a
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Physiological Studies of Lactococcus lactis
DEFF Research Database (Denmark)
Hansen, Gunda
medium and (specific) acidification activity in milk in response to the extracellular pH (pHex) during batch fermentations and in response to stress during downstream processing and storage as frozen, freeze- and spray-dried cells. In this PhD thesis, in situ flow cytometric viability assessment...... fermentation, downstream processing and storage in the absence of a protectant. However, storing freeze-dried L. lactis cells at 30 °C negatively affected the culturability and acidification activity. The reactivation of freeze-dried cells in fermentation medium prior to flow cytometric viability assessment...
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Assessment of RELAP5-3D copyright using data from two-dimensional RPI flow tests
International Nuclear Information System (INIS)
Davis, C.B.
1998-01-01
The capability of the RELAP5-3D copyright computer code to perform multi-dimensional thermal-hydraulic analysis was assessed using data from steady-state flow tests conducted at Rensselaer Polytechnic Institute (RPI). The RPI data were taken in a two-dimensional test section in a low-pressure air/water loop. The test section consisted of a thin vertical channel that simulated a two-dimensional slice through the core of a pressurized water reactor. Single-phase and two-phase flows were supplied to the test section in an asymmetric manner to generate a two-dimensional flow field. A traversing gamma densitometer was used to measure void fraction at many locations in the test section. High speed photographs provided information on the flow patterns and flow regimes. The RPI test section was modeled using the multi-dimensional component in RELAP5-3D Version BF06. Calculations of three RPI experiments were performed. The flow regimes predicted by the base code were in poor agreement with those observed in the tests. The two-phase regions were observed to be in the bubbly and slug flow regimes in the test. However, nearly all of the junctions in the horizontal direction were calculated to be in the stratified flow regime because of the relatively low velocities in that direction. As a result, the void fraction predictions were also in poor agreement with the measured values. Significantly improved results were obtained in sensitivity calculations with a modified version of the code that prevented the horizontal junctions from entering the stratified flow regime. These results indicate that the code's logic in the determination of flow regimes in a multi-dimensional component must be improved. The results of the sensitivity calculations also indicate that RELAP5-3D will provide a significant multi-dimensional hydraulic analysis capability once the flow regime prediction is improved
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Collective flow in 2.76 and 5.02 A TeV Pb + Pb collisions
Energy Technology Data Exchange (ETDEWEB)
Zhao, Wenbin; Xu, Hao-jie [Peking University, Department of Physics and State Key Laboratory of Nuclear Physics and Technology, Beijing (China); Song, Huichao [Peking University, Department of Physics and State Key Laboratory of Nuclear Physics and Technology, Beijing (China); Collaborative Innovation Center of Quantum Matter, Beijing (China); Peking University, Center for High Energy Physics, Beijing (China)
2017-09-15
In this paper, we study and predict flow observables in 2.76 and 5.02 A TeV Pb + Pb collisions, using the iEBE-VISHNU hybrid model with TRENTo and AMPT initial conditions and with different forms of the QGP transport coefficients. With properly chosen and tuned parameter sets, our model calculations can nicely describe various flow observables in 2.76 A TeV Pb + Pb collisions, as well as the measured flow harmonics of all charged hadrons in 5.02 A TeV Pb + Pb collisions. We also predict other flow observables, including v{sub n}(p{sub T}) of identified particles, event-by-event v{sub n} distributions, event-plane correlations, (normalized) symmetric cumulants, non-linear response coefficients and p{sub T}-dependent factorization ratios, in 5.02 A TeV Pb + Pb collisions. We find many of these observables to remain approximately the same values as the ones in 2.76 A TeV Pb + Pb collisions. Our theoretical studies and predictions could shed light to the experimental investigations in the near future. (orig.)
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Flow cytometric analysis of cell killing by the jumper ant venom peptide pilosulin 1.
King, M A; Wu, Q X; Donovan, G R; Baldo, B A
1998-08-01
Pilosulin 1 is a synthetic 56-amino acid residue polypeptide that corresponds to the largest allergenic polypeptide found in the venom of the jumper ant Myrmecia pilosula. Initial experiments showed that pilosulin 1 lysed erythrocytes and killed proliferating B cells. Herein, we describe how flow cytometry was used to investigate the cytotoxicity of the peptide for human white blood cells. Cells were labeled with fluorochrome-conjugated antibodies, incubated with the peptide and 7-aminoactinomycin D (7-AAD), and then analyzed. The effects of varying the peptide concentration, serum concentration, incubation time, and incubation temperature were measured, and the cytotoxicity of pilosulin 1 was compared with that of the bee venom peptide melittin. The antibodies and the 7-AAD enabled the identification of cell subpopulations and dead cells, respectively. It was possible, using the appropriate mix of antibodies and four-color analysis, to monitor the killing of three or more cell subpopulations simultaneously. We found that 1) pilosulin 1 killed cells within minutes, with kinetics similar to those of melittin; 2) pilosulin 1 was a slightly more potent cytotoxic agent than melittin; 3) both pilosulin 1 and melittin were more potent against mononuclear leukocytes than against granulocytes; and 4) serum inhibited killing by either peptide.
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Grosset, Andrée-Anne; Loayza-Vega, Kevin; Adam-Granger, Éloïse; Birlea, Mirela; Gilks, Blake; Nguyen, Bich; Soucy, Geneviève; Tran-Thanh, Danh; Albadine, Roula; Trudel, Dominique
2017-12-21
Hematoxylin and eosin (H&E) staining is a well-established technique in histopathology. However, immunohistochemistry (IHC) interpretation is done exclusively with hematoxylin counterstaining. Our goal was to investigate the potential of H&E as counterstaining (H&E-IHC) to allow for visualization of a marker while confirming the diagnosis on the same slide. The quality of immunostaining and the fast-technical performance were the main criteria to select the final protocol. We stained multiple diagnostic tissues with class I IHC tests with different subcellular localization markers (anti-CK7, CK20, synaptophysin, CD20, HMB45, and Ki-67) and with double-staining on prostate tissues with anti-high molecular weight keratins/p63 (DAB detection) and p504s (alkaline phosphatase detection). To validate the efficacy of the counterstaining, we stained tissue microarrays from the Canadian Immunohistochemistry Quality Control (cIQc) with class II IHC tests (ER, PR, HER2, and p53 markers). Interobserver and intraobserver concordance was assessed by κ statistics. Excellent agreement of H&E-IHC interpretation was observed in comparison with standard IHC from our laboratory (κ, 0.87 to 1.00), and with the cIQc reference values (κ, 0.81 to 1.00). Interobserver and intraobserver agreement was excellent (κ, 0.89 to 1.00 and 0.87 to 1.00, respectively). We therefore show for the first time the potential of using H&E counterstaining for IHC interpretation. We recommend the H&E-IHC protocol to enhance diagnostic precision for the clinical workflow and research studies.
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The use of flow cytometry to examine calcium signalling by TRPV1 in mixed cell populations.
Assas, Bakri M; Abdulaal, Wesam H; Wakid, Majed H; Zakai, Haytham A; Miyan, J; Pennock, J L
2017-06-15
Flow cytometric analysis of calcium mobilisation has been in use for many years in the study of specific receptor engagement or isolated cell:cell communication. However, calcium mobilisation/signaling is key to many cell functions including apoptosis, mobility and immune responses. Here we combine multiplex surface staining of whole spleen with Indo-1 AM to visualise calcium mobilisation and examine calcium signaling in a mixed immune cell culture over time. We demonstrate responses to a TRPV1 agonist in distinct cell subtypes without the need for cell separation. Multi parameter staining alongside Indo-1 AM to demonstrate calcium mobilization allows the study of real time calcium signaling in a complex environment. Copyright © 2017. Published by Elsevier Inc.
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Fluid flow in 0.5-m scale blocks of Topopah Spring tuff
International Nuclear Information System (INIS)
Blair, S. C.; Carlson, S. R.; Constantino, M. S.
1999-01-01
A laboratory experiment was conducted on a 0.5-m scale block of Topopah Spring tuff, to measure fluid flow and mechanical deformation properties under conditions that approximate the near-field environment of a potential nuclear waste repository, and to provide an intermediate-scale test case for numerical model validation. The test specimen is a 0.25 x 0.25 x 0.50 m rectangular prism bisected by an artificial (saw-cut) fracture orthogonal to the tuff fabric. Water was supplied by a point source at the center of the fracture under various pressures of up to 0.04 MPa. Both fluid flow and mechanical properties were found to be anisotropic and strongly correlated with the ash flow fabric. Fluid mass-balance measurements revealed that only minor imbibition of water occurred through the fracture surfaces and that flow rates were independent of normal stress to 14.0 MPa and temperature to 140 C. Flow through the fracture occurred largely through uncorrelated porosity that intersected the fracture plane
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Assessment of critical flow models of RELAP5-MOD2 and CATHARE codes
International Nuclear Information System (INIS)
Hao Laomi; Zhu Zhanchuan
1992-01-01
The critical flow tests for the long and short nozzles conducted on the SUPER MOBY-DICK facility were analyzed using the RELAP5-MOD2 and CATHARE 1.3 codes to assess the critical flow models of two codes. The critical mass flux calculated for two nozzles are given. The CATHARE code has used the thermodynamic nonequilibrium sound velocity of the two-phase fluid as the critical flow criterion, and has the better interphase transfer models and calculates the critical flow velocities with the completely implicit solution. Therefore, it can well calculate the critical flowrate and can describe the effect of the geometry L/D on the critical flowrate
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Chowdhury, Pankaj; Gomaa, Hassan; Ray, Ajay K
2015-02-01
In this paper, we have studied Eosin Y-sensitized sacrificial hydrogen generation with triethanolamine as electron donor in UV, visible, and solar light irradiation. Aeroxide TiO2 was loaded with platinum metal via solar photo-deposition method to reduce the electron hole recombination process. Photocatalytic sacrificial hydrogen generation was influenced by several factors such as platinum loading (wt%) on TiO2, solution pH, Eosin Y to Pt/TiO2 mass ratio, triethanolamine concentration, and light (UV, visible and solar) intensities. Detailed reaction mechanisms in visible and solar light irradiation were established. Oxidation of triethanolamine and formaldehyde formation was correlated with hydrogen generation in both visible and solar lights. Hydrogen generation kinetics followed a Langmuir-type isotherm with reaction rate constant and adsorption constant of 6.77×10(-6) mol min(-1) and 14.45 M(-1), respectively. Sacrificial hydrogen generation and charge recombination processes were studied as a function of light intensities. Apparent quantum yields (QYs) were compared for UV, visible, and solar light at four different light intensities. Highest QYs were attained at lower light intensity because of trivial charge recombination. At 30 mW cm(-2) we achieved QYs of 10.82%, 12.23% and 11.33% in UV, visible and solar light respectively. Copyright © 2014 Elsevier Ltd. All rights reserved.