Complex Interactions between Dioxin-Like and Non-Dioxin-Like Compounds for in Vitro Cellular Responses: Implications for the Identification of Dioxin Exposure Biomarkers

Despite considerable advances in reducing the production of dioxin-like toxicants in recent years, contamination of the food chain still occasionally occurs resulting in huge losses to the agri-food sector and risk to human health through exposure. Dioxin-like toxicity is exhibited by a range of stable and bioaccumulative compounds including polychlorinated dibenzo-p-dioxins (PCDDs) and dibenzofurans (PCDFs), produced by certain types of combustion, and man-made coplanar polychlorinated biphenyls (PCBs), as found in electrical transformer oils. While dioxinergic compounds act by a common mode of action making exposure detection biomarker based techniques a potentially useful tool, the influence of co-contaminating toxicants on such approaches needs to be considered. To assess the impact of possible interactions, the biological responses of H4IIE cells to challenge by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) in combination with PCB-52 and benzo-a-pyrene (BaP) were evaluated by a number of methods in this study. Ethoxyresorufin-O-deethylase (EROD) induction in TCDD exposed cells was suppressed by increasing concentrations of PCB-52, PCB-153, or BaP up to 10 mu M. BaP levels below 1 mu M suppressed TCDD stimulated EROD induction, but at higher concentrations, EROD induction was greater than the maximum observed when cells were treated with TCDD alone. A similar biphasic interaction of BaP with TCDD co-exposure was noted in the AlamarBlue assay and to a lesser extent with PCB-52. Surface enhanced laser desorption/ionization-time of flight mass spectrometry (SELDI-TOF) profiling of peptidomic responses of cells exposed to compound combinations was compared. Cells co-exposed to TCDD in the presence of BaP or PCB-52 produced the most differentiated spectra with a substantial number of non-additive interactions observed. These findings suggest that interactions between dioxin and other toxicants create novel, additive, and non-additive effects, which may be more indicative of the types of responses seen in exposed animals than those of single exposures to the individual compounds.

Model Calculations and Implementation of Filters and Hybrid Green VCSELs based on Optical Thin Film Stacks

This work introduced the novel conception of complex coupled hybrid VCSELs for the first time. Alternating organic and inorganic layers in the lasers provide periodic variation of refractive index and optical gain, which enable single mode operation and low threshold of the VCSELs. Model calculations revealed great reduction of the lasing threshold with factors over 30, in comparison with the existing micro-cavity lasers. Tunable green VCSEL has been also designed, implemented and analyzed taking advantage of the broad photoluminescence spectra of the organics. Free standing optical thin films without compressive stress are technologically implemented. Multiple membrane stacks with air gap in between have been fabricated for the implementation of complex coupled VCSEL structures. Complex coupled hybrid VCSEL is a very promising approach to fill the gaps in the green spectral range of the semiconductor lasers.

Fluorescent AlPO(4) gels and glasses doped with Rhodamine 6G: Preparation, structural and optical characterization

Fluorescent AlPO(4) xerogels doped with different amounts of Rhodamine 6G (Rh6G) laser dye were prepared by a one-step sal-gel process. In addition, mesoporous AlPO(4) glasses obtained from undoped gels were loaded with different amounts of Rh6G by wet impregnation. Optical excitation and emission spectra of both series of samples show significant dependences on Rh6G concentration, revealing the influence of dye molecular aggregation. At comparable dye concentrations the aggregation effects are found to be significantly stronger in the gels than in the mesoporous glasses. This effect might be attributed to stronger interactions between the dye molecules and the glass matrix, resulting in more efficient dye dispersion in the latter. The interaction of Rh6G with the glassy AlPO(4) network has been probed by (27)Al and (31)P solid-state NMR techniques. New five- and six-coordinated aluminum environments have been observed and characterized by advanced solid-state NMR techniques probing (27)Al-(1)H and (27)Al-(31)P internuclear dipole couplings. The fractional area of these new Al sites is correlated with the combined fractional area of two new Q(3Al)((0)) and Q(2Al)((0)) phosphate species observed in the (31)P MAS NMR spectra. Based on this correlation as well as detailed composition dependent studies, we suggest that the new signals arise from the breakage of Al-O-P linkages associated with the insertion process. (C) 2010 Elsevier B.V. All rights reserved.

Morphological regional differences of epithelial cells along the midgut in Diatraea saccharalis Fabricius (Lepidoptera : Crambidae) larvae

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Preparação e caracterização óptica de filmes nanocristalinos de GaAs:H depositados por RF magnetron sputtering

Pós-graduação em Ciência e Tecnologia de Materiais - FC

Estudo da adesão dentinária em restaurações realizadas com ionômero modificado por resina: influência de técnicas restauradoras e do preparo cavitário

Pós-graduação em Ciência Odontólogica - FOA

A Data-Analytic Strategy for Protein-Biomarker Discovery: Profiling of High-Dimensional Proteomic Data for Cancer Detection

With recent advances in mass spectrometry techniques, it is now possible to investigate proteins over a wide range of molecular weights in small biological specimens. This advance has generated data-analytic challenges in proteomics, similar to those created by microarray technologies in genetics, namely, discovery of "signature" protein profiles specific to each pathologic state (e.g., normal vs. cancer) or differential profiles between experimental conditions (e.g., treated by a drug of interest vs. untreated) from high-dimensional data. We propose a data analytic strategy for discovering protein biomarkers based on such high-dimensional mass-spectrometry data. A real biomarker-discovery project on prostate cancer is taken as a concrete example throughout the paper: the project aims to identify proteins in serum that distinguish cancer, benign hyperplasia, and normal states of prostate using the Surface Enhanced Laser Desorption/Ionization (SELDI) technology, a recently developed mass spectrometry technique. Our data analytic strategy takes properties of the SELDI mass-spectrometer into account: the SELDI output of a specimen contains about 48,000 (x, y) points where x is the protein mass divided by the number of charges introduced by ionization and y is the protein intensity of the corresponding mass per charge value, x, in that specimen. Given high coefficients of variation and other characteristics of protein intensity measures (y values), we reduce the measures of protein intensities to a set of binary variables that indicate peaks in the y-axis direction in the nearest neighborhoods of each mass per charge point in the x-axis direction. We then account for a shifting (measurement error) problem of the x-axis in SELDI output. After these pre-analysis processing of data, we combine the binary predictors to generate classification rules for cancer, benign hyperplasia, and normal states of prostate. Our approach is to apply the boosting algorithm to select binary predictors and construct a summary classifier. We empirically evaluate sensitivity and specificity of the resulting summary classifiers with a test dataset that is independent from the training dataset used to construct the summary classifiers. The proposed method performed nearly perfectly in distinguishing cancer and benign hyperplasia from normal. In the classification of cancer vs. benign hyperplasia, however, an appreciable proportion of the benign specimens were classified incorrectly as cancer. We discuss practical issues associated with our proposed approach to the analysis of SELDI output and its application in cancer biomarker discovery.

Protein profiling of organic stone matrix and urine from dogs with urolithiasis

Two-thirds of the organic matrix in urinary stones consists of proteins. Their relationship to calculogenesis remains controversial with regard to their effect as inhibitors or promoters during stone formation. The purpose of the present study was to determine the differences in peptide and protein pattern between the urine of stone formers (n = 23) and control dogs (n = 12), as well as between organic matrix of different urinary stones (struvite n = 11, calcium oxalate n = 8, uric acid n = 4) using surface-enhanced laser desorption/ionization time-of-flight mass spectrometry. Specific differences in protein and peptide profiles were found in the organic matrix of different mineral compositions. Characteristic differences were also found in urinary peptide and protein pattern especially in molecular masses below 20 kDa between affected and healthy dogs. Based on the obtained molecular masses they were in some cases tentatively identified as proteins that are known to be involved in stone formation in humans. The study shows that in dogs, specific-urinary peptides and proteins might be associated with urolithiasis. It indicates the importance to further characterize those proteins for possible diagnostic purposes in prognosis and therapy

Influence of hormone replacement therapy on proteomic pattern in serum of postmenopausal women

OBJECTIVES: Proteomics approaches to cardiovascular biology and disease hold the promise of identifying specific proteins and peptides or modification thereof to assist in the identification of novel biomarkers. METHOD: By using surface-enhanced laser desorption and ionization time of flight mass spectroscopy (SELDI-TOF-MS) serum peptide and protein patterns were detected enabling to discriminate between postmenopausal women with and without hormone replacement therapy (HRT). RESULTS: Serum of 13 HRT and 27 control subjects was analyzed and 42 peptides and proteins could be tentatively identified based on their molecular weight and binding characteristics on the chip surface. By using decision tree-based Biomarker Patternstrade mark Software classification and regression analysis a discriminatory function was developed allowing to distinguish between HRT women and controls correctly and, thus, yielding a sensitivity of 100% and a specificity of 100%. The results show that peptide and protein patterns have the potential to deliver novel biomarkers as well as pinpointing targets for improved treatment. The biomarkers obtained represent a promising tool to discriminate between HRT users and non-users. CONCLUSION: According to a tentative identification of the markers by their molecular weight and binding characteristics, most of them appear to be part of the inflammation induced acute-phase response

Protein profiling of urine from dogs with renal disease using ProteinChip analysis

Measurement of total urinary proteins in individuals that tested positive by urinary dipstick is a typical method for assessing the presence of potentially serious renal disorders. In the absence of such overt proteinuria, however, measurement of specific urinary proteins may be useful in the diagnosis of nephropathies and may provide greater insight into the pathogenesis. The urine of 28 dogs (16 with renal disease and 12 healthy) was evaluated to determine whether specific low-molecular-weight proteins or the pattern of protein excretion could also be used as a marker of tubular dysfunction in dogs. Specific proteins were assessed by immunological methods, whereas protein profiles were determined by surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (MS). In particular, changes in the excretion of retinol-binding protein (RBP) and Tamm-Horsfall protein (THP) appear to be of clinical relevance in the diagnosis of canine kidney diseases. The pattern of urinary protein and peptides revealed specific changes in abundance in dogs with renal disease at molecular masses (kD) of 11.58, 12.41, 12.60, 14.58, 20.95 (RBP), 27.85, and 65.69 (albumin). In conclusion, comparable proteins as in humans might be used as urinary markers for proximal (RBP) and distal (THP) tubular dysfunction in dogs. Surface-enhanced laser desorption/ionization time-of-flight MS is a promising tool for the study of kidney physiology and pathophysiology and might aid in the discovery of new biomarkers of renal disease.

Zooplankton abundance and size structure in the North Atlantic from MSM26_133-14

Data on zooplankton abundance and biovolume were collected in concert with data on the biophysical environment at 9 stations in the North Atlantic, from the Iceland Basin in the East to the Labrador Sea in the West. The data were sampled along vertical profiles by a Laser Optical Plankton Counter (LOPC, Rolls Royce Canada Ltd.) that was mounted on a carousel water sampler together with a Conductivity-Temperature-Depth sensor (CTD, SBE19plusV2, Seabird Electronics, Inc., USA) and a fluorescence sensor (F, ECO Puck chlorophyll a fluorometer, WET Labs Inc., USA). Based on the LOPC data, abundance (individuals/m**3) and biovolume (mm3/m**3) were calculated as described in the LOPC Software Operation Manual [(Anonymous, 2006), http://www.brooke-ocean.com/index.html]. LOPC data were regrouped into 49 size groups of equal log10(body volume) increments, see Edvardsen et al. (2002, doi:10.3354/meps227205). LOPC data quality was checked as described in Basedow et al. (2013, doi:10.1016/j.pocean.2012.10.005). Fluorescence was roughly converted into chlorophyll based on filtered chlorophyll values obtained from station 10 in the Labrador Sea. Due to the low number of filtered samples that was used for the conversion the resulting chlorophyll values should be considered with care. CTD data were screened for erroneous (out of range) values and then averaged to the same frequency as the LOPC data (2 Hz). All data were processed using especially developed scripts in the python programming language. The LOPC is an optical instrument designed to count and measure particles (0.1 to 30 mm equivalent spherical diameter) in the water column, see Herman et al., (2004, doi:10.1093/plankt/fbh095). The size of particles as equivalent spherical diameter (ESD) was computed as described in the manual (Anonymous, 2006), and in more detail in Checkley et al. (2008, doi:10.4319/lo.2008.53.5_part_2.2123) and Gaardsted et al. (2010, doi:10.1111/j.1365-2419.2010.00558.x).

Zooplankton abundance and size structure in the North Atlantic from MSM26_135-21

Data on zooplankton abundance and biovolume were collected in concert with data on the biophysical environment at 9 stations in the North Atlantic, from the Iceland Basin in the East to the Labrador Sea in the West. The data were sampled along vertical profiles by a Laser Optical Plankton Counter (LOPC, Rolls Royce Canada Ltd.) that was mounted on a carousel water sampler together with a Conductivity-Temperature-Depth sensor (CTD, SBE19plusV2, Seabird Electronics, Inc., USA) and a fluorescence sensor (F, ECO Puck chlorophyll a fluorometer, WET Labs Inc., USA). Based on the LOPC data, abundance (individuals/m**3) and biovolume (mm3/m**3) were calculated as described in the LOPC Software Operation Manual [(Anonymous, 2006), http://www.brooke-ocean.com/index.html]. LOPC data were regrouped into 49 size groups of equal log10(body volume) increments, see Edvardsen et al. (2002, doi:10.3354/meps227205). LOPC data quality was checked as described in Basedow et al. (2013, doi:10.1016/j.pocean.2012.10.005). Fluorescence was roughly converted into chlorophyll based on filtered chlorophyll values obtained from station 10 in the Labrador Sea. Due to the low number of filtered samples that was used for the conversion the resulting chlorophyll values should be considered with care. CTD data were screened for erroneous (out of range) values and then averaged to the same frequency as the LOPC data (2 Hz). All data were processed using especially developed scripts in the python programming language. The LOPC is an optical instrument designed to count and measure particles (0.1 to 30 mm equivalent spherical diameter) in the water column, see Herman et al., (2004, doi:10.1093/plankt/fbh095). The size of particles as equivalent spherical diameter (ESD) was computed as described in the manual (Anonymous, 2006), and in more detail in Checkley et al. (2008, doi:10.4319/lo.2008.53.5_part_2.2123) and Gaardsted et al. (2010, doi:10.1111/j.1365-2419.2010.00558.x).

Zooplankton abundance and size structure in the North Atlantic from MSM26_127-22

Data on zooplankton abundance and biovolume were collected in concert with data on the biophysical environment at 9 stations in the North Atlantic, from the Iceland Basin in the East to the Labrador Sea in the West. The data were sampled along vertical profiles by a Laser Optical Plankton Counter (LOPC, Rolls Royce Canada Ltd.) that was mounted on a carousel water sampler together with a Conductivity-Temperature-Depth sensor (CTD, SBE19plusV2, Seabird Electronics, Inc., USA) and a fluorescence sensor (F, ECO Puck chlorophyll a fluorometer, WET Labs Inc., USA). Based on the LOPC data, abundance (individuals/m**3) and biovolume (mm3/m**3) were calculated as described in the LOPC Software Operation Manual [(Anonymous, 2006), http://www.brooke-ocean.com/index.html]. LOPC data were regrouped into 49 size groups of equal log10(body volume) increments, see Edvardsen et al. (2002, doi:10.3354/meps227205). LOPC data quality was checked as described in Basedow et al. (2013, doi:10.1016/j.pocean.2012.10.005). Fluorescence was roughly converted into chlorophyll based on filtered chlorophyll values obtained from station 10 in the Labrador Sea. Due to the low number of filtered samples that was used for the conversion the resulting chlorophyll values should be considered with care. CTD data were screened for erroneous (out of range) values and then averaged to the same frequency as the LOPC data (2 Hz). All data were processed using especially developed scripts in the python programming language. The LOPC is an optical instrument designed to count and measure particles (0.1 to 30 mm equivalent spherical diameter) in the water column, see Herman et al., (2004, doi:10.1093/plankt/fbh095). The size of particles as equivalent spherical diameter (ESD) was computed as described in the manual (Anonymous, 2006), and in more detail in Checkley et al. (2008, doi:10.4319/lo.2008.53.5_part_2.2123) and Gaardsted et al. (2010, doi:10.1111/j.1365-2419.2010.00558.x).