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PubMed1993

Mutagenicity of organic extracts from Santiago (Chile) airborne particulate matter.

The Ames test has been used to detect the mutagenic activity of organic extracts from Santiago (Chile) airborne particles collected in 1990 and 1991 in one of the monitoring net system stations (MACAM). The samples were assayed with the strains TA98, TA98-NR, and TA98/1,8-DNP6 of Salmonella typhimurium, in the presence and in the absence of liver S9 fraction obtained from rats treated with Aroclor 1254. With the strain TA98 all the samples showed a very high mutagenic response either in the presence or in the absence of S9 fraction, suggesting that Santiago airborne particles contain both indirect-acting (polycyclic aromatic hydrocarbons) and direct-acting mutagenic agents. The mutagenicity of Santiago airborne particles was much higher than that reported in studies performed in other countries. Results obtained with the strains TA98-NR and TA98/1,8-DNP6 suggest that the extracts also contain mononitro and dinitroarenes. These nitroarenes have been described as very potent mutagenic agents, that can be generated by photochemical reactions under certain atmospheric conditions, or in the combustion of fuel, especially of diesel motors. The presence of polycyclic aromatic hydrocarbons and nitroarenes in Santiago airborne particles, as well as the high levels of mutagenicity detected, suggest that the inhabitants permanent exposure to these kinds of compounds represents a high risk for human health.

Air Pollutants↗

PubMed1993

31P NMR examination of phosphorus metabolites in the aqueous, acidic, and organic extracts of Phaseolus vulgaris seeds.

31P NMR spectroscopy was used to compare the phosphorus compound content in aqueous, acidic, and organic extracts of Phaseolus vulgaris seeds. Two chloroform-methanol and three ethanol extractions were used to isolate phospholipids from dry cotyledons and the phospholipid profiles were compared. Variations in phospholipid composition and artificially high concentrations of lysophosphatidylcholine were observed in the ethanol extracts. Other phosphorus compounds were extracted using perchloric acid, trichloroacetic acid, trichloroacetic acid in ether, hydrochloric acid, boiling water, and aqueous Hepes. The Hepes extraction was introduced in order to compare the results of a gentle procedure with those involving heat and acid treatments. Low concentrations of phospho-sugars and other phosphorus metabolites were found in all these extracts. High concentrations of phytate were found in all aqueous and acidic extractions; however, 31P NMR spectra of aqueous extractions did not show the phytate resonance. The NMR silence of the phytate resonances is attributed to complexation of phytate. The 31P NMR spectra of aqueous extracts contained a broad resonance at 0.1 ppm assigned to protein-bound RNA. Various extraction procedures on the same biological material are presented, and the comparison of these methods will facilitate future analysis and interpretation of literature reports involving these methods of tissue analysis.

Chemistry Techniques, Analytical↗

PubMed1990

Application of a subtraction hybridization technique involving photoactivatable biotin and organic extraction to solution hybridization analysis of genomic DNA.

We have adapted a subtraction hybridization technique involving photoactivatable biotin, streptavidin binding, and organic extraction for solution hybridization analysis of mammalian genomic DNA. By combining maximal hybridization conditions of high salt, dextran sulfate, and formamide with successive hybridization steps and sequence enrichment by agarose gel electrophoresis, up to 97% of tracer DNA can be reproducibly driven to hybridize with photobiotinylated driver DNA. We demonstrate that the fractionation of hybridized from unhybridized sequences by this technique differs from hydroxyapatite chromatography with respect to the handling of nondenatured tracer, foldback sequences, and tracer-tracer hybrids. Strategies are presented to control the contribution of these species to the final subtracted product thereby making this technology a useful adjunct to solution hybridization approaches such as deletion cloning.

Azides↗

PubMed2005

Characterization of water extractable organic matter in a deep soil profile.

The aim of this study was to identify qualitative and quantitative differences of water extractable organic matter (WEOM) isolated from each horizon along a deep soil profile and to evaluate any relationship between the WEOC and the total organic carbon (TOC) content. The soil profile "Monte Pietroso" is located in the Murge area, Apulia region in Southern Italy. Samples from the eight horizons (Ap1, Ap2, Ab1, Ab2, Bt1, 2B, 2Bt2, and 2B/C) were collected in October 2002. The WEOM characterization was carried out by means of UV absorbance, fluorescence spectroscopy in the emission and excitation/emission matrix (EEM) modes, and additional spectroscopic derived indexes. Soil organic carbon was shown to accumulate in the top horizons (Ap) and, in general, to decrease with depth, whereas, the WEOM/TOC ratio increases with increasing depth. The aromaticity and the humification index of the WEOM decrease dramatically downward the soil profile, whereas the fluorescence efficiency index tends to increase markedly. The WEOM fractions feature three main fluorophores with different wavelength and relative intensity. In general WEOM transport phenomena are suggested to occur downward the soil profile, depending on the nature of the organic material and on the chemical and mineral characteristics of the various horizons.

Italy↗

PubMed2005

Seasonal and spatial variation of solvent extractable organic compounds in fine suspended particulate matter in Hong Kong.

The results of a 12-month study of more than 100 solvent extractable organic compounds (SEOC) in particulate matter (PM) less than or equal to 2.5 microm (PM2.5) collected at three air monitoring stations located at roadside, urban, and rural sites in Hong Kong are reported. The total yield of SEOC that accounts for approximately 8-18% of organic carbon (OC) determined by a thermal optical transmittance method was 125-2060 ng/m3, which included 14.6-128 ng/m3 resolved aliphatic hydrocarbons, 39.4-1380 ng/m3 unresolved complex mixtures, 0.6-17.2 ng/m3 polycyclic aromatic hydrocarbons, 41.6-520 ng/m3 fatty acids, and < 0.1-12.1 ng/m3 alkanols. Distinct seasonal variations (summer/winter differences) were observed with higher concentrations of the total and each class of SEOC in the winter and lower concentrations in the summer. Spatial variations are also obvious, with the roadside samples having the highest concentrations of SEOC and the rural samples having the lowest concentrations in all seasons. Characteristic ratios of petroleum hydrocarbons, such as carbon preference index, unresolved to resolved components, and carbon number with maximum concentration, suggest that PM2.5 carbon in Hong Kong originates from both biogenic and anthropogenic sources. The proportion of SEOC in PM2.5 from anthropogenic sources is estimated.

Air Pollutants↗

PubMedDate not supplied

Antiproliferative effects of an organic extract from the marine diatom Skeletonema costatum (Grev.) Cleve. Against a non-small-cell bronchopulmonary carcinoma line (NSCLC-N6).

An organic extract of the marine diatom Skeletonema costatum was studied in vitro for its effect on asynchronous cells of a human non-small-cell bronchoplumanory carcinoma line (NSCLC-N6). Cell growth appeared to be inhibited in the G1 phase of the cell cycle, and kinetic studies in pretreated cells showed that this growth arrest was irreversible. These events are related to a terminal maturation induction.

Antineoplastic Agents↗

PubMed1998

The organ extraction and splanchnic haemodynamic effects of octreotide in cirrhotic patients.

BACKGROUND: Intravenous octreotide is an established treatment of oesophageal variceal haemorrhage in the cirrhotic patient. AIM: To examine the organ extraction and splanchnic haemodynamic effects of octreotide in cirrhotic patients with portal hypertension. METHODS: Thirteen patients with cirrhosis had hepatic venous catheterization performed. Hepatic venous pressure gradient (HVPG), indocyanine green (ICG) clearance and hepatic blood flow (HBF) were determined in the basal state and during 60 min of octreotide infusion by bolus injection (0.75 microg/kg) followed by continuous infusion of 0.75 microg/kg x h. Blood samples were simultaneously drawn from the femoral artery and the hepatic and renal veins. RESULTS: The extraction fraction of octreotide in the liver was 0.05 (-0.01 - 0.14) (median (interquartile range)) and in the kidneys 0.16 (-0.06 - 0.35). The extraction fraction ratio (E(liver)/E(kidney)) was 0.69 (-0.20 - 1.06). Hepatic clearance was 47 mL/min (3-88) (n = 11). No correlations were found between liver biochemistry or galactose elimination capacity (GEC; a metabolic measure of liver function) and renal extraction fraction or liver clearance. Octreotide had no effect on HVPG or wedged hepatic venous pressure although free hepatic venous pressure increased during octreotide infusion: 6 mmHg (5-9) vs. 7 mmHg (6-10) (P = 0.02). No effect on HBF was observed while ICG clearance decreased significantly. CONCLUSIONS: Octreotide is extracted in cirrhotic patients by both the liver and the kidney, the latter being the most important organ of elimination. Octreotide decreases liver metabolic activity determined by the ICG clearance technique, but no significant effects of octreotide on HVPG or HBF could be demonstrated.

Adult↗

PubMed1997

Mutagenicity testing of organic extracts of diesel exhaust particles after fractionation and recombination.

A new strategy for the evaluation of mixtures is presented. The mixture used was the organic extract of diesel exhaust particles (DEP). After extraction with dichloromethane (DCM), the crude extract was fractionated according to polarity into five fractions: aliphatic hydrocarbons, polycyclic aromatic hydrocarbons (PAHs), nitro-PAHs, dinitro-PAHs, and polar compounds. After dissolving in dimethylsulphoxide (DMSO), the three fractions containing the primary mutagens (fractions 3-5) were recombined in different combinations to create new extracts. The blend matrix was obtained using a mixture design at three dose levels to support an empirical model with linear, interaction, and quadratic terms (Taylor polynome). The recombined extracts were tested in the Ames Salmonella assay using strain TA100. Multivariate data analysis was performed with projections to latent structures (PLS). The best model describing the relation between the mutagenicity (response) and the three fractions (variables) contained two interaction terms. The model showed high correlation (r2) and prediction properties (Q2), the latter obtained after cross validation. Interaction terms are only indications of possible synergism or antagonism and have to be evaluated with respect to dose-additivity and response-additivity. The incorporation of dose in the design reduced the number of samples (recombined extracts) significantly, compared to determining dose-response curves on each sample (i.e. the recombined extracts in different dilutions). Furthermore, instead of running two independent experiments as required in the standard procedure for the Ames test, predictions and verifications of a few new samples were used. The principle of fractionation and recombination, and the use of mixture design may in principle be extended to an unlimited number of variables. An adaptation of mixture design to the isobole method is discussed.

Hydrocarbons↗

PubMed1990

Single-step organic extraction of leukotrienes and related compounds and their simultaneous analysis by high-performance liquid chromatography.

A method for the simultaneous single-step organic extraction from biological matrices of peptido- and dihydroxyleukotrienes as well as 5-hydroperoxy- and 5-hydroxyeicosatetraenoic acid followed by separation and quantitation in a single run on reversed-phase high-performance liquid chromatography was evaluated. Using an extraction system comprising 400/1200/4800 (v/v/v) aqueous phase/isopropanol/dichloromethane, pH 3.0, absolute recoveries of 82.3 +/- 2.0, 89.7 +/- 1.0, 93.7 +/- 1.4, 92.8 +/- 1.4, 90 +/- 4, and 90 +/- 4% for prostaglandin B1 (PGB1), leukotriene C4 (LTC4), leukotriene B4 (LTB4), leukotriene D4 (LTD4), 5-hydroperoxyeicosatetraenoic acid (5-HETE), respectively, were achieved. Separation and quantitation of products were performed on a Nucleosil 100 C18 column (5 microns, 4.6 X 250 mm) using, at pH 6.0, a gradient system comprising 72/28/0.02 (v/v/v) methanol/water/glacial acetic acid from 0 to 15 min, followed by a convex gradient to 76/24/0.02 (v/v/v) methanol/water/glacial acetic acid, followed by a 10-min hold at this methanol concentration. The method was used to investigate the profile of leukotrienes synthesized by rat hepatocyte homogenates from 5-HPETE or leukotriene A4 in absence or presence of glutathione (GSH). During a 5-min incubation with 100 microM 5-HPETE, 9.6 ng LTB4/mg protein and 2.2 micrograms 5-HETE/mg protein were formed in the absence of GSH. In the presence of 0.4 mM GSH, 3.7 ng LTB4/mg protein and 11.0 micrograms 5-HETE/mg protein were formed. Using 20 microM LTA4 as a substrate, 17.3 and 324.0 ng LTC4/mg protein X min and 14.3 and 19.3 ng LTB4/mg protein X min were formed in the presence of 0.4 and 10 mM GSH, respectively.

1-Propanol↗

PubMedDate not supplied

Relationship between compost stability and extractable organic carbon.

Establishing a simple yet reliable compost stability test is essential for a better compost quality control and utilization efficiency. The objective of this study was to examine the relationship between extractable organic carbon (OC) and compost stability based on 18 compost samples from five composting facilities. The compost samples were extracted sequentially with water for 2 h [water(2h)] and 0.1 M NaOH for 2 and 24 h [NaOH(2h) and NaOH(24h), respectively]. The extractable OC was further separated into fulvic acid (FA) and humic acid (HA) fractions by adjusting the pH to <2. The mass specific absorbance (MSA) of OC in the six fractions was measured. Compost stability was estimated with a CO2 evolution method. The extractable OC concentration was influenced by the total volatile solids and decreased with curing time for compost with a high level of extractable OC. The OC levels in each fraction were significantly correlated (p < 0.05) to each other except for the water(2h)-extractable HA. In addition, all the FA and HA fractions except for water(2h)-extractable HA were highly (P < 0.01) and linearly correlated to CO2 evolution, but multiple regression showed that NaOH(24h)-extractable OC was insignificant for CO2 evolution. The relatively high slope of NaOH(2h)-extractable FA versus CO2 evolution suggests that this fraction may contribute the most to compost CO2 evolution. The water(2h)- and/or NaOH(2h)-extractable FA tests are recommended for measuring compost stability because of their high correlation with CO2 evolution. This estimation can be obtained through a simple photometric method covering a wide range of carbon concentrations up to 4,000 mg L(-1).

Carbon↗

PubMed1986

Experimental systemic amyloidosis induced by immunization with syngeneic organ extracts in mice.

Systemic amyloidosis was induced consistently in mice by intramuscular injection of syngeneic organ (liver and kidney) extracts mixed with CFA six times at weekly intervals. Syngeneic organ extract with CFA also induced amyloidosis of a lesser degree. All three strains of mice (C57BL/6, C3H/He, and BALB/c) injected with a syngeneic liver extract mixed with CFA developed systemic amyloidosis; the most prominent amyloid deposition occurred in C57BL/6 (B6) mice, followed by C3H/He and BALB/c. The amyloid substance deposited in these animals was identified as mouse amyloid A protein (AA). Furthermore, an organ specificity of the immunogen in inducing amyloidosis was suggested with liver and kidney extracts. Primed spleen cells of the immunized B6 mice were fractionated by a nylon-wool column and injected to normal recipient mice via the tail vein. Organs of the recipient mice developed systemic amyloidosis 8 wk after the transfer, and the most prominent histological changes occurred in the recipient mice given nylon-wool column adherent spleen cells. Using anti-Thy-1,2; Ly-1; Ly-2, antibody and complement, it was suggested that T cells, especially Ly-1,2,3+ T cell populations in the primed nylon-wool adherent cells, play an important role in the induction of systemic amyloidosis. It was shown further that the amyloidosis-inducing substance in liver extract was composed of unstable proteins or protein-bound substance.

Amyloidosis↗

PubMed2002

Genotoxic activity of extractable organic matter from urban airborne particles in Shanghai, China.

The aim of this research is to investigate the impact of air pollution on the population in Shanghai. The genotoxicity of extractable organic matter (EOM) from the air particles was investigated by the means of the Salmonella plate incorporation assay, rat hepatocyte unscheduled DNA repair assay, and mice micronuclei test. The airborne particles were collected in 13 locations during the summer of 1992 and winter of 1993. The crude extracts were fractionated by acid-base partitioning into acid, base and neutral fractions. The neutral fractions were further fractionated by resin-silica gel column chromatography into three subfractions. The induction of revertants with the crude extracts was higher in winter samples than in summer samples. Both indirect-acting and direct-acting mutagenicity were observed. The mutagenicity was detected with TA98, but was not detected with TA100. The mutagenic activity was the greatest in the acid, aromatic and polar fractions from summer samples. The fractions from the winter samples did not show clear differences. There was no substantial location-related variance in the mutagenic potencies of EOM, but substantial location- or time-related variances in the mutagenic potencies of the airborne particles per cubic meter air were found. While rat hepatocyte unscheduled DNA synthesis (UDS) assay revealed genotoxicity for all the samples, there was no big variance in the genotoxicity of the fractions. The mouse micronuclei test showed results similar to the UDS assay. The difference of locality did not have statistical significance.

Air Pollutants↗

PubMed1997

[Studies of malignant transformation of human fetal lung cells induced by organic extract from tap water].

Expression of P53 protein was determined in the process and the end of induction of malignant transformation in human fetal lung cells, with organic extract from tap water in a river of City G, to study its molecular mechanism. Results showed that the water sample of experimental doses could induce formation of cell malignant transformation foci to different extent and had certain toxicity to cells. High expression of P53 protein was observed in cell nuclei 15 days after exposure and before formation of cell transformation foci, or 30 days after exposure and formation of them. It suggests that changes of P53 protein may relate to cell malignant transformation to certain extent, and play some roles in the process of it.

Cell Transformation, Neoplastic↗

PubMed1984

A broadly active viral inhibitor in human and animal organ extracts and body fluids.

To help assess the possibility that a newly described viral inhibitor from cell cultures might play a natural defensive role in vivo, its distribution and concentration in human and animal organ extracts and body fluids were investigated. The concentration of the inhibitor was high in human liver, heart muscle, splenic extracts, and human serum and milk. The inhibitor in the body was indistinguishable from a previously described inhibitor produced in cell cultures that was characterized by broad antiviral activity, lack of target cell species specificity, lack of induction of stable antiviral activity in cells, rapid reversibility of antiviral action, prevention of virus attachment, and stability at 100 degrees C. Sixteen virus plaque reduction units of the inhibitor diminished the yield of poliovirus in vitro by more than 1000-fold. Additional evidence that contact-blocking viral inhibitor (CVI) inhibits vaccinia virus attachment to cells is presented. A role for the inhibitor in natural defense against viral infections is possible.

Animals↗

PubMed1983

Phenotypic plasticity of cultured bovine chromaffin cells. I. Morphological changes induced by non-chromaffin cells and organ extracts, but not by mouse and bovine nerve growth factor.

Adult bovine chromaffin cells are known to show process outgrowth in culture, similar to that seen with rat chromaffin and pheochromocytoma cells after exposure to nerve growth factor (NGF). To determine whether bovine chromaffin cells respond to NGF or NGF-like factors, dissociated adrenal medullary cells from adult cattle were cultured for up to 4 weeks in the presence or absence of NGF (from mouse submaxillary glands and bovine seminal vesicles, respectively), adrenal non-chromaffin cells and various organ extracts. Chromaffin cells that had been freed from non-chromaffin cells by differential plating and/or gamma-irradiation showed virtually no fiber outgrowth. In the presence of adrenal non-chromaffin cells (NCC), cell-free extracts obtained from these cells or medium conditioned by them, chromaffin cells formed processes. Extracts from bovine seminal vesicles (SVE), but not from other organs including bovine brain, heart, liver, kidney, adrenal glands and male mouse submaxillary glands, also elicited fiber outgrowth. Extension of processes induced by NCC and SVE could not be blocked by administration of monospecific anti-NGF antibodies directed against NGF from mouse submaxillary glands. Purified NGF from mouse submaxillary glands and SVE did not elicited a response. We conclude that adult bovine chromaffin cells in culture show structural plasticity similar to that shown by cultured chromaffin cells from other species, but do not respond to NGF. Neurite outgrowth promoting activities appear to reside with macromolecular constituents of bovine adrenal non-chromaffin cells and SVE.

Animals↗

PubMed2003

Fast fractionation of complex organic extracts by normal-phase chromatography on a solid-phase extraction polymeric sorbent. Optimization of a method to fractionate wine flavor extracts.

Some properties of LiChrolut-EN resins as normal-phase sorbent have been studied. Retention factors using pentane as solvent range from less than 2 (ethyl esters) to more than 56 (fatty acids and vanillin). All retention factors were smaller than 2 with dichloromethane. The efficiency of the bed was between 4 and 13 plates per cm. A method for the fast fractionation of wine flavor extracts has been further developed. Wine (75 ml) is extracted on a 0.5 g LiChrolut-EN bed. Volatile compounds are recovered in 5 ml of dichloromethane and the extract is further concentrated to 0.1 ml. Recoveries of the extraction procedure are above 85% for all compounds less polar than isoamyl alcohol. This extract is fractionated on a bed (5.0 cm height, 0.6 cm internal diameter) packed with 0.55 g of LiChrolut-EN resins. A first fraction is collected by the elution with 4 ml of pentane. A second one with 6 ml of a mixture pentane/dichloromethane (9:1) and a final fraction with 4 ml of dichloromethane. The first fraction is enriched in ethyl esters and some other non-polar compounds. The second fraction concentrates the alcohols and some volatile phenols, while the third is enriched in fatty acids, vanillin derivatives and some lactones. The recovery in the fractionation is complete. The profile obtained in the fractionation is very stable, and becomes distorted only when the column is loaded with an extract containing 80 mg of major volatiles (coming from more than 150 ml of wine). The fractionation of extracts from different wines showed that the performance of the process does not depend on the composition of the extract. Twenty-seven out of 32 studied compounds eluted reproducibly mainly in one fraction. The results suggest that the method can be applied as an aid for qualitative or quantitative analysis to any kind of organic extract as an alternative to liquid chromatography on silica-gel.

Chromatography, Liquid↗

PubMed1998

Mutagenicity testing of organic extracts of diesel exhaust particles after spiking with polycyclic aromatic hydrocarbons (PAH).

In the present study, spiking was used as a strategy to evaluate the mutagenicity of individual compounds in a mixture. Mutagenicity of individual polycyclic aromatic hydrocarbons (PAH) was evaluated in an organic extract of diesel exhaust particles (DEP). The particles were extracted with dichloromethane (DCM). After replacing DCM with dimethylsulphoxide (DMSO), the extract was spiked with four individual PAH: benzo(a)pyrene, benzo(a)anthracene, pyrene and fluoroanthene. The PAH were added separately and in various combinations to the extract to determine the effects of each variable and to identify possible interactions between the individual PAH and between the PAH and the extract. The study was designed as a fractional factorial experiment with the five variables (the DEP extract and the four PAH), giving 16 (instead of 32) mixtures plus a triplicate centrepoint and background, i.e. a total of 20. The fractionated factorial design used in the present work supports a model with linear and interaction terms. The mixtures were tested for mutagenicity in the Ames assay using four strains of Salmonella typhimurium in the presence of rat liver xenobiotic enzymes (S9-mix). Projections to Latent Structures (PLS) was used to quantify the mutagenicity of each compound and possible interactions. The four individual PAH and the DEP extract acted additively in the Ames test with 10% S9-mix.

Anthracenes↗

PubMed2005

Impact of meteorology and energy structure on solvent extractable organic compounds of PM2.5 in Beijing, China.

Twenty-eight PM2.5 samples collected in Summer (July 2002) and Winter (November 2002) at two sites in Beijing, China were analyzed using GC/MS to investigate the impact of meteorology and coal burning on the solvent extractable organic compounds (SEOC). The characteristics and abundance of the n-alkanes, polycyclic aromatic hydrocarbons (PAHs), n-fatty acids and n-alkanols were determined. Source identification was made using organic species as molecular markers. Semi-volatile compounds of alkanes and PAHs had much higher concentrations in winter than summer because of the large difference in the temperature between the seasons. Plant wax emission was a major contributor to n-alkanes in summer, but fossil fuel residue was a major source (>80%) in winter. The seasonal differences in the distribution of pentacyclic triterpanes clearly shows the impact of coal burning for space heating in winter. The yield of PAHs in winter (148 ng m(-3) at the urban site and 277 ng m(-3) at the suburban site) was six to eight times higher than that in summer and was found to be mainly from coal burning. Higher pollutant concentrations were measured at the suburban site than the urban site in winter due to the rapid expansion of the city limit and the relocation of factories from urban to suburban areas over the last two decades.

Aerosols↗