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The antipsoriatic compound anthralin influences bioenergetic parameters and redox properties of energy transducing membranes.

Bioenergetic parameters and redox properties of energy transducing membranes in rat liver mitochondria and cyanobacteria were investigated in the presence of the antipsoriatic compound anthralin (1,8-dihydroxy-9-anthrone). Transmembrane pH and electrical gradients were determined using electron paramagnetic resonance spectroscopy. In mitochondria, ubiquinones 9,10 and other redox components of the electron transport chain are reduced by anthralin; the proton motive force is increased. In the absence of ADP, anthralin slightly stimulates mitochondrial cyanide-insensitive oxygen consumption. It is suggested that increased cyanide-insensitive respiration is due to enhanced autoxidation of mitochondrial components and/or catalyzed oxidation of anthralin. In the presence of ADP mitochondrial respiration is decreased, and ATP synthesis is inhibited. Uncoupler-induced mitochondrial respiration is also decreased by anthralin, indicating inhibition of the electron transport chain. In the cyanobacterium Synechococcus PCC 6311 anthralin increases the pH gradient and decreases ATP levels. Thus, anthralin acts as an electron donor to membrane associated redox components and inhibits ATP synthesis in two different biologic systems. In human keratinocytes oxygen metabolism is influenced by anthralin in a similar pattern as in isolated mitochondria, and ATP content is decreased. Because anthralin reacts with redox components in different biologic membranes, alterations of subcellular/cellular redox status and energy metabolism might contribute significantly to its antiproliferative activity.

Animals↗

Anopheles gambiae feeding and survival on honeydew and extra-floral nectar of peridomestic plants.

It is widely believed that the malaria vector Anopheles gambiae Giles (Diptera: Culicidae) rarely or never feeds on sugar in nature. If so, the need for supplemental blood-feeding may be increased and this would help to explain why it is such an efficient malaria vector. Nonetheless, both sexes of this mosquito species readily imbibe and digest sugar solutions, and sugar is a staple of laboratory colonies. In this study, we investigated whether An. gambiae will feed on the extra-floral nectar of three common peridomestic plants in Africa, and on honeydew of the mealybug Pseudococcus longispinus (Targioni-Tozetti) (Hemiptera: Homoptera: Pseudococcidae), and how this affects survivorship. We found that both males and females of An. gambiae provided with vegetative parts of cassava (Manihot esculenta Crantz) survived as well (x = 26.3 and 19.2 days, respectively) as they did on 50% sucrose solution (x = 29.7 and 24.3 days, respectively) and much longer than they did on water alone (x = 1.8 days, both sexes). Females provided with mealybug honeydew also lived substantially longer (x = 16.5 days) than those on water alone. Males and females provided with vegetative parts of castorbean (Ricinus communis L.) also survived much longer (x = 12.7 and 7.8 days, respectively) than on water, but those provided with flowering lantana (Lantana camara L.) did not. Anthrone tests of females after one night of exposure to these potential energy sources confirmed that they obtained fructose from cassava, from mealybug honeydew, and from non-flowering castorbean, but not from lantana or from castorbean lacking its petiolar nectaries. Previous laboratory studies had shown that sugar availability affects the survival and biting frequency of An. gambiae. It now appears that this mosquito can locate natural sources of plant sugar readily and utilize them effectively. Nectar-producing plants in the domestic environment may play a significant role in this mosquito's energy budget and malaria vectorial capacity.

Animals↗

Feeding and survival of the malaria vector Anopheles gambiae on plants growing in Kenya.

The propensity of the malaria vector mosquito Anopheles gambiae Giles (Diptera: Culicidae) to ingest sugars from various plants, and subsequent survival rates, were assessed with laboratory-reared males and females offered eight species of plants commonly cultivated and/or growing wild in western Kenya. In cages (no-choice bioassay), mosquitoes given the opportunity to feed on castorbean (Ricinus communis L.) had the longest survival times (mean and median survival time of 6.99 +/- 0.23 and 5.67 +/- 0.17 days, respectively), comparable to mosquitoes given 6% glucose (mean and median survival time of 8.70 +/- 0.23 and 6.67 +/- 0.33 days, respectively). Survival rates of An. gambiae were low on the other plants, comparable to mosquitoes given only water. Three plants: sweet potato (Ipomoea batatas L.), wild sage (Lantana camara L.) and castorbean provided levels of sugar ingestion by both sexes of An. gambiae detectable using the cold anthrone method, showing a positive correlation between median survival and sugar consumption (Spearman rank correlation coefficient = 0.905, P < 0.0001). Equal numbers of males and females were released in an enclosed semi-field screenhouse system containing a range of local plants, but no host for blood, and allowed to feed ad libitum: 6.7 +/- 0.5% (11/64) of those recaptured were found to contain detectable fructose (all females). Common plants are clearly a viable source of nutrition for adult female An. gambiae, as well as males, and may constitute and important resource for this important malaria vector.

Animals↗

Biological activities of endotoxins from Yersinia enterocolitica.

The chemical properties and the general biological activities of lipopolysaccharide (LPS) and Boivin-type endotoxin obtained respectively by phenol-water and trichloroacetic acid extraction from Yersinia enterocolitica serotypes O3 and O9 were studied. The yield of LPS from the O9 strain was about 10% of the O3 strain possibly because of the lower solubility of O9-LPS in aqueous phase. However, the chemical composition of O9-LPS was similar to that of O3-LPS in the proportions of reducing sugar, glucosamine, heptose, KDO, and lipid A. In pyrogenicity and local Shwartzman reactivity in rabbits and lethality for mice, there was also no difference between O3 and O9-LPS. The anthrone-positive carbohydrate and lipid A contents of Boivin-type endotoxin from O3 were higher than those of the endotoxin from O9. The biological activities of Boivin-type endotoxin from O3 were also remarkably higher than those of the endotoxin from O9. It seems that endotoxin of Y. enterocolitica serotype O3 may play an important role in infection by this organism.

Adjuvants, Immunologic↗

Hydroxyethyl starch. An agent for hypovolaemic schock treatment II. Urinary excretion in normal volunteers following three consecutive daily infusions.

1 Urinary hydroxyethyl starch (HES) concentrations were determined by the anthrone method, in four healthy normal male volunteers following three consecutive daily 500 ml infusions (total 1500 ml), in order to ascertain excretion rates under normal controlled conditions. 2 The HES was excreted at a rate of 2.57, 2.46, and 2.44 g/h during the first hour postinjection, on days 1, 2, and 3, respectively. 3 The rate during the interval 12-24 h postinjection, averaged 0.14, 0.21, and 0.10 g/h, on days 1,2, and 3, respectively. 4 In the intervals 24-72, 72-120, and 120-168 h after the third and final injection, the excretion rate was 0.07, 0.04, and 0.02 g/h, respectively. 5 Renal function, as assessed by serum creatinine and uric acid concentrations and 24 h void volumes, was normal during the entire period of observation. 6 The results indicate that HES is excreted at a similar rate following three consecutive daily infusions without evidence of renal injury.

Adult↗

Nectar feeding by mosquitoes in Sweden, with special reference to Culex pipiens and Cx torrentium.

1. Nectar feeding by mosquitoes collected from tansy (Tanacetum vulgare) flowers was studied in July and August 1983-85 at two sites in central and south-western Sweden. Prior to fructose analysis, gonotrophic state and parity was determined. 2. A total of 1010 mosquitoes (70% males) of eighteen species was collected. Fructose was detected in 75% of the males and 78% of the females. Most (86%) mosquitoes collected belonged to Culex pipiens and Cx torrentium. 3. Among 219 Cx pipiens females, for most of which the abdomen appeared gravid (52%) or empty (42%), 81% were fructose-positive. Females of other species were also proved to have fed on plant sugars in both early and later stages of the gonotrophic cycle. 4. In Cx pipiens and males of Cx torrentium peak nectar feeding occurred between 22.00 and 04.00 hours. Data for males of both species suggested a V-shaped pattern of nectar feeding activity during the night. 5. At about 16 degrees C nearly all mosquitoes became fructose-negative in the anthrone test within 20 h after collection from tansy flowers. 6. At least 44% of Cx pipiens females that were attracted to a dove-baited trap had fed on plant sugars shortly before.

Animals↗

Leishmaniasis in the Jordan Valley. V. Dispersal characteristics of the sandfly Phlebotomus papatasi.

Several characteristics of dispersing and non-dispersing Phlebotomus papatasi (Scopoli) were quantified and compared. The majority of dispersing sandflies, trapped crossing fallow fields, were females (68.5% v. 51.1%); of the dispersing females, 55.4% were parous, 48.1% were inseminated and 11.6% were gravid. In the population of sandflies sampled exiting from burrows of the sand rat Psammomys obesus Cretschmar, these categories, respectively, represented 39%, 90% and 26% of the females examined. Leishmania promastigotes were found in 9% of females exiting from P. obesus burrows, and in 2.7% of the dispersing females. The anthrone test established that the reason for activity of gravid females is sugar feeding. These females do not disperse and are suitable targets for future control measures.

Animals↗

Sugar meal sources for the phlebotomine sandfly Lutzomyia longipalpis in Ceará State, Brazil.

In the Brazilian village of Boqueirão do Renato Parente, Ceará State, where visceral leishmaniasis is endemic, significant variation in the abundance of phlebotomine sandflies between different types of vegetation was demonstrated by castor oil sticky trap collections. Population densities of Lutzomyia longipalpis (Lutz & Neiva) sandflies were higher in beans and maize crops than in the surrounding open and tree scrub. Significant relationships were detected between the abundances of sandflies and aphids, suggesting aphid honeydew as a potential food source. Sugar meal analysis, using high-performance anion exchange (HPAE) chromatography with pulsed amperometric detection (PAD) and the cold anthrone test, was used to demonstrate that significantly more sandflies fed on bean plants contaminated with aphids and honeydew than on uncontaminated beans. Furthermore, higher concentrations of sugars were detected in flies which fed on aphid/honeydew-contaminated beans, suggesting that sugar was more easily available and/or that honeydew/aphids act as phagostimulants for sandflies. Amongst wild sandflies collected from a bean field, significantly more female sandflies were 'sugar-positive' than males, though the sugar concentrations in positive sandflies were similar for both sexes. The concentrations of di- and trisaccharides in sandfly homogenates, honeydew extracts and phloem exudates were very low.

Animals↗

Agar accumulates in rat peritoneal macrophages elicited with thioglycollate broth.

Thioglycollate broth- and agar-elicited rat peritoneal exudate cells contain copious amounts of agar, a sulphated polysaccharide contained in the eliciting agent; the agar content has been measured chemically by the anthrone method for neutral polysaccharides. Adherence experiments confirm that day-3 thioglycollate-elicited macrophages contain 95% of the measured agar; on day 1, 72% is associated with polymorphonuclear cells. Agar ingestion by macrophages coincides with increases in glycogen synthase activity. We suggest that ingestion of agar may be related to the reported deficiencies in function of thioglycollate-elicited or -treated macrophages.

Agar↗

Changes in the molecular composition of circulating hydroxyethyl starch.

1. Hydroxyethyl starch (HES) of molecular weight 450,000 has been extensively investigated as a plasma volume expander and HES of molecular weight 60,000 has been proposed for use as a cryoprotective agent in the freezing for preservation of red blood cells. In these applications considerable amounts of HES may be injected intravenously and it is therefore desirable to understand its behaviour in vivo.2. After intravenous injection into rabbits, plasma levels of HES and clinical dextrans were determined using anthrone. HES persisted longer in the circulation than did the dextrans.3. HES was recovered from the bloodstream of rabbits and shown by gel filtration to be of a narrower molecular size distribution than the injected material. Smaller molecules were removed, presumably by filtration at the glomerulus. Larger molecules may have been removed by cells of the reticuloendothelial system or possibly through the action of amylase.4. The amylase in saliva was shown to break down HES into molecules of intermediate size with very little production of maltose units.

Amylases↗

Resolution of liver fibrosis in chronic CCl4 administration in the rat after discontinuation of treatment: effect of silymarin, silibinin, colchicine and trimethylcolchicinic acid.

The purpose of this work was to obtain a suitable model of fibrosis, in which spontaneous reversion was minimal, to study the ability of silymarin, silibinin, colchicine and trimethylcolchicinic acid (TMCA) to reverse it. Reversal of liver fibrosis was studied in male Wistar rats after one, two or three months of CCl(4) administration (0.4 g/kg intraperitoneally, three times per week), by discontinuation of the toxin for 2 months. Silymarin (50 mg/kg), silibinin (50 mg/kg), colchicine (10 microg/rat) and trimethylcolchicinic acid (100 microg/rat) were administered daily, by gavage, after 3 months of CCl(4) administration. Collagen content was determined by measuring hydroxyproline in liver samples; glycogen, was determined utilizing the anthrone reagent; Mallory's trichromic stains of liver sections were performed. The best scheme of treatment was obtained when CCl(4) was administered during three months (collagen increased 6 times). Discontinuation of the toxin for two months produced a significant but relative small reduction of fibrosis (collagen was still 4.5 times over control). Colchicine, TMCA, silymarin or silibinin treatment showed no significant fibrolitic effect. This scheme of treatment may be an excellent tool to study the ability of drugs to reverse fibrosis. The hepatoprotective properties of silymarin, silibinin, colchicine and trimethylcolchinic acid may be irrelevant to reverse established cirrhosis.

Alanine Transaminase↗

Anthralin-derived transients--I. The triplet state and the products of its reaction with oxygen in benzene.

Direct laser excitation in benzene of 1,8-dihydroxy-9-anthrone (anthralin) does not lead to transient species with lifetimes in the nanosecond time regime or longer. The triplet state has been produced in benzene by pulse radiolysis and characterised in terms of its absorption spectrum (lambda max 560 nm), natural lifetime (11 microseconds), self-quenching properties (kmicrosecond = 2.6 x 10(7) l mol-1 s-1) and triplet energy (234 kJ mol-1). There is no tendency in the non-polar medium for production of either the triplet or ground state in a tautomeric form. The observed triplet state reacts with oxygen with a typical rate constant, 2.2 x 10(9) l mol-1 s-1. The products of this reaction are singlet oxygen (approximately 64%) and the anthralin radical (approximately 14%).

Anthralin↗

Anthralin-derived transients--II. Formation of the radical by spontaneous fragmentation of both singlet and triplet states of the 10,10'-dehydrodimer: radical pair multiplicity effects.

The singlet and triplet states of the anthralin (1,8-dihydroxy-9-anthrone) dehydrodimer have been produced selectively in benzene via pulsed laser excitation and pulse radiolysis respectively. The lifetime of S1 is less than or equal to 30 ps, that of T1 short but unspecified. Both states fragment spontaneously to yield a pair of anthralin radicals. The singlet radical pair predominantly undergoes geminate recombination within the solvent cage. In contrast, the corresponding triplet radical pair undergoes essentially exclusive cage escape to give the anthralin free radical (lambda max 370, 490 and 720 nm) which recombines under normal diffusive conditions. Both recombination processes lead, at least in part, to one or more species which have been assigned as tautomeric forms of the original dimer. The anthralin free radical in benzene is insensitive to the vitamin E model 6-hydroxy-2,2,5,7,8-pentamethylchroman and reacts only slowly with oxygen.

Anthralin↗

Chemical Characterization of Polysaccharide from the Slime Layer of the Cyanobacterium Microcystis flos-aquae C3-40.

Macromolecular material from the slime layer of the cyanobacterium Microcystis flos-aquae C3-40 was defined as material that adhered to cells during centrifugation in growth medium but was dislodged by washing with deionized water and retained within dialysis tubing with a molecular-weight cutoff of 3,500. At each step of this isolation procedure, the slime was observed microscopically. Cells in the centrifugal pellet were surrounded by large amounts of slime that excluded negative stain, whereas cells that had been washed with water lacked visible slime. Two independently isolated lots of slime contained no detectable protein (<1%, wt/wt) and consisted predominantly of anthrone-reacting polysaccharide. Sugars in a hydrolysate of slime polysaccharide were derivatized with trimethylsilylimidazole and examined by gas chromatography-mass spectrometry. The composition of the slime polysaccharide was 1.5% (wt/wt) galactose, 2.0% glucose, 3.0% xylose, 5.0% mannose, 5.5% rhamnose, and 83% galacturonic acid. This composition resembles that of the plant polysaccharide pectin, which was treated in parallel as a control. Consistent with earlier indications that M. flos-aquae slime preferentially binds certain cations, the ratio of Fe to Na in the dialyzed slime was 10 times that in the growth medium. The composition of the slime is discussed with respect to possible mechanisms of cation binding in comparison with other cyanobacterial exopolysaccharides and pectin.

Journal Article↗

Selection of a highly monensin-resistant Prevotella bryantii subpopulation with altered outer membrane characteristics.

Prevotella bryantii cultures treated with monensin grew more slowly than untreated cultures, but only if the monensin concentration was greater than 1 microM. Cultures that were repeatedly transferred (eight transfers or 25 doublings) with monensin always grew rapidly, even at a 10 microM concentration. The amount of monensin needed to facilitate half-maximal potassium depletion (K(d)) from monensin-selected cells was 16-fold greater than "unadapted" wild-type cultures (3,200 versus 200 nM). Cells taken from continuous culture had a K(d) of 100 nM, and these inocula could not grow in batch culture when the monensin concentration was greater than 300 nM. Continuous cultures treated with monensin nearly washed out, but the surviving cells had a K(d) of 1,300 nM. When wild-type cells were transferred in batch culture with 10 microM monensin, the K(d) did not reach its maximum value (3,200 nM) until after eight transfers (25 doublings). K(d) declined when monensin was removed, and it took eight transfers to reach the control value (200 nM). The most probable number of wild-type cells was 1,000-fold lower than of the monensin-selected cells, but calculations based on relative growth advantage and K(d) indicated that the wild-type culture had 1 to 10% highly monensin-resistant cells. Cell pellets of wild-type cultures were more difficult to disperse than were monensin-selected cells, and water-soluble phenol extracts of monensin-selected cells had 1.8-fold more anthrone-reactive material than did the wild type. Wild-type cultures that were washed in Tris buffer (pH 8.0) released little alkaline phosphatase and were agglutinated by lysozyme. Monensin-selected cultures leaked ninefold more alkaline phosphatase and were not agglutinated by lysozyme. Wild-type colonies taken from high-dilution agar roll tubes retained the lysozyme agglutination phenotype even if transferred with monensin, and monensin-selected colonies were never agglutinated. These observations indicated that wild-type P. bryantii cultures had a subpopulation with different outer membrane characteristics and increased monensin resistance.

Adenosine Triphosphate↗

Effect of herbicides on cellulose decomposition by Sporocytophaga myxococcoides.

The effects of 14 herbicides on cellulose decomposition by Sporocytophaga myxococcoides were studied by growing the organism in two different liquid cellulose-mineral salts media in the presence of the test agent and then quantitatively determining the residual cellulose. In the first medium, containing acid-hydrolyzed cellulose, anthrone reagent was used to determine residual carbohydrate. In the second medium, Gooch Pyrex fritted-glass crucibles were used to determine the amount of residual Whatman powdered cellulose. The herbicides used were Atrazine, Simazine, Dacthal, Diuron, Amiben, Banvel-D, Banvel-T,2,3,6-TBA, Dicryl, Maleic Hydrazide, Stam F-34, Zytron, Fenac, and Dalapon. Of these, Zytron appeared to be the most inhibitory to the test bacterium, and was the only compound exhibiting inhibition at a commonly used field rate of application (2 ppm).

Bacteria↗

Transient response of continuously cultured heterogeneous populations to changes in temperature.

Completely mixed, once-through continuous culture systems of heterogeneous microbial populations of sewage origin were systematically examined for response to changes in reactor temperature. Systems were operated at two dilution rates of 0.125 and 0.25 per h. "Steady state" conditions of the systems were assessed with the reactors operating at 25 C. From this base line, temperature was decreased to as low as 8 C and increased to as high as 57.5 C. Response was assessed in the ensuing transient phase as the system approached a new "steady state." The response was measured by changes in amount and type of carbon source in the reactor effluent as determined by the chemical oxygen demand test, the anthrone test, and gas chromatography. Biological solids concentration and cell composition (protein, carbohydrate, ribonucleic acid and deoxyribonucleic acid) were also determined. These systems responded more favorably to increases than to decreases in temperature. Regardless of the direction of change, the system with the lowest dilution rate (D = 0.125 per h) responded more successfully; i.e., there was less leakage of carbon source in the effluent and less dilute-out of cells during the transient phase.

Bacteria↗

Prostaglandin E release from human monocytes treated with lipopolysaccharides isolated from Bacteroides intermedius and Salmonella typhimurium: potentiation by gamma interferon.

The purpose of this investigation was to examine gamma interferon potentiation of lipopolysaccharide (LPS) responses in human monocytes by using phenol-water-extracted (unfractionated) and highly purified LPS preparations isolated from Bacteroides intermedius and Salmonella typhimurium. Phenol-water-extracted LPS preparations from these bacteria were further purified by chromatography over Sepharose-CL-4B. LPS enrichment in pooled column fractions was assessed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and quantitation of hydroxy-fatty acid and 2-keto-3-deoxyoctulosonic acid content, protein contamination, and anthrone-reactive material. Monocyte stimulation by LPS, measured as prostaglandin E (PGE) release, was assessed with and without gamma interferon treatment. Cells were either treated simultaneously with gamma interferon and LPS or pretreated with gamma interferon prior to LPS stimulation. PGE release from counterflow-isolated monocytes was quantitated during the 0- to 24-h and 24- to 48-h culture intervals. Contrary to previous results from this laboratory, phenol-water-extracted LPS preparations from B. intermedius and S. typhimurium were similar in their capacities to stimulate PGE release from monocytes. Molecular sieve chromatography was found to remove substantial amounts of high-molecular-weight polysaccharide contaminants only from the B. intermedius LPS but did not significantly alter the potency of either B. intermedius or S. typhimurium LPS. Gamma interferon cotreatment did not potentiate the release of PGE with any of the LPS preparations tested. However, 24-h pretreatment of monocytes with gamma interferon followed by a 24-h exposure to LPS resulted in significant potentiation of PGE release over LPS alone. In addition, B. intermedium preparations were approximately threefold more potent than similarly prepared LPS isolates from S. typhimurium following gamma interferon pretreatment. These results indicate that gamma interferon can selectively potentiate the effects of B. intermedius LPS in human monocyte isolates.

Bacteroides↗