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PubMed1993

Automated enzymatic analysis of inulin.

We have developed an automated enzymatic assay for quantitation of inulin in plasma and urine that can be performed on the Cobas FARA II. In the assay, inulinase hydrolyzes inulin to fructose, and sorbitol dehydrogenase converts fructose to sorbitol with consumption of NADH, which is detected by spectrophotometry. The method incorporates a sample blank (inactivated inulinase) for each specimen to subtract contributions of endogenous fructose. Recovery of fructose or inulin was near 100%, with linearity to 300 mg/L. The enzymatic assay (y) agreed well with an anthrone comparison method (x) for analysis of inulin in both urine specimens (y = 1.00x - 138; Sy/x = 714) and plasma specimens (y = 1.00x - 3.5; Sy/x = 5.5). Glucose at 300 mg/L yielded an apparent inulin value of 1.3 mg/L in the enzymatic assay, but reacted at nearly 10% equivalency in the anthrone assay. Interferences from sorbitol, mannitol, and xylitol were negligible. CVs for day-to-day precision studies were 1-4%. The automated enzymatic assay of inulin is faster and avoids the use of caustic reagents required by the classical anthrone method.

Anthracenes↗
PubMed2005

Antimalarial compounds from Kniphofia foliosa roots.

During the course of screening Ethiopian medicinal plants for their antimalarial properties, it was found that the dichloromethane extract of the roots of Kniphofia foliosa Hochst. (Asphodelaceae), which have long been used in the traditional medicine of Ethiopia for the treatment of abdominal cramps and wound healing, displayed strong in vitro antiplasmodial activity against the chloroquine-sensitive 3D7 strain of Plasmodium falciparum with an ED50 value of 3.8 microg/mL and weak cytotoxic activity against KB cells with an ED50 value of 35.2 microg/mL. Five compounds were isolated from the roots and evaluated for their in vitro antimalarial activity. Among the compounds tested, 10-(chrysophanol-7'-yl)-10-(xi)-hydroxychrysopanol-9-anthrone and chryslandicin, showed a high inhibition of the growth of the malaria parasite, P. falciparum with ED50 values of 0.260 and 0.537 microg/mL, respectively, while the naphthalene derivative, 2-acetyl-1-hydroxy-8-methoxy-3-methylnaphthalene, exhibited a less significant antimalarial activity with an ED50 value of 15.4 microg/mL. To compare the effect on the parasite with toxicity to mammalian cells, the cytotoxic activities of the isolated compounds against the KB cell line were evaluated and 10-(chrysophanol-7'-yl)-10-(xi)-hydroxychrysopanol-9-anthrone and chryslandicin displayed very low toxicity with ED50 values of 104 and 90 microg/mL, respectively. This is the first report of the inhibition of the growth of P. falciparum by anthraquinone-anthrone dimers and establishes them as a new class of potential antimalarial compounds with very little host cell toxicity.

Animals↗
PubMed1989

Evaluation in vitro and in vivo of two labelling techniques of different 99mTc-dextrans for lymphoscintigraphy.

Five dextrans with different molecular weights and charges were labelled with 99mTc. The labelling methods presented by Henze et al. (1982a) and Ercan et al. (1985) were compared. The labelling efficiency was tested with gel column chromatography scanning (GCS), gel chromatography (GC) combined with the Anthrone test, paper chromatography (PC) and thin layer chromatography (TLC). The GCS technique always indicated a lower labelling efficiency than the PC and TLC techniques, which was due to a more optimal separation of the radioactive components. Gel chromatography in combination with the Anthrone test made it easy to identify the different radiochemical components in contrast to the other methods. Dextran solutions were injected subcutaneously bilaterally at the xiphoid processes in rabbits. The injection sites were massaged for 30 s. Uptake in the parasternal lymph nodes was registrated with a scintillation camera. The animals were killed and dissected at the end of the study. This investigation shows that the labelling method of Ercan et al. gives the highest labelling efficiency. Furthermore, the final pH (4.5) for the dextran solution makes it more useful for injection. For quality control of 99mTc-labelled dextran we recommend the Anthrone test as a complement to GC because it is a quick and simple method of determining the dextran content.

Animals↗
PubMed1982

[The elimination of hydroxyethyl starch 200/0.5, dextran 40 and oxypolygelatine (author's transl)].

After withdrawal of 400 ml whole blood and subsequent infusion of 500 ml of a colloidal plasma substituent, the intravascular and renal colloid elimination was investigated in 40 test subjects. The individual colloidal solutions could no longer be demonstrated in the intravascular space after the following times: 10% hydroxyethyl starch 200/0.5 (anthrone method) after six weeks, 10% dextran 40 (anthrone method) after two weeks, 6% hydroxyethyl starch 200/0.5 (anthrone method) after four weeks and 5.5% oxypolygelatine (hydroxyproline method) after two days. Colloidal plasma substitutes are polydisperse solutions with various molecular weights and degree of hydroxyethylation and therefore, also have a large number of different elimination constants. With repeated application, the intravascular colloid concentration shifts in favour of the molecules with a longer half life which are difficult to eliminate. The elimination of the clinically employed dextran 40 and oxypolygelatine solution could be best described with an open two-compartment model. As a result of its greater heterogeneity, the elimination of the moderately high molecular weight hydroxyethyl starch 200/0.5 could only be characterized approximately even assuming three elimination constants. In the first four days, the hydroxyethyl starch 200/0.5 was more rapidly eliminated compared to dextran 40. However, subsequently a very much lower elimination from the intravascular space was found for about 3% of the administered hydroxyethyl starch 200/0.5. Oxypolygelatine was eliminated especially rapidly. Accordingly, the greatest renal clearance was found for oxypolygelatine, which showed a close relation to the molecular weight. On the other hand, a rapid elimination simultaneously is followed by a correspondingly lower volume effect.

Adult↗
PubMed1990

Kinetic studies on anthralin photooxidation.

The photooxidation of the antipsoriatic drug anthralin (1,8-dihydroxy-9-anthrone) has been studied by several kinetic techniques, including direct observation of 1O2 (1 delta g) luminescence at 1.27 microns. The rate of deactivation of 1O2 increases at higher pH, demonstrating that the trihydroxyanthracene anion is the reactive species. Direct determination of the rate constant of 1O2 deactivation (kR + kQ) in deuterated buffer systems by luminescence quenching gave a value of 3.0 x 10(8) M-1 s-1 for the anion; the neutral anthrone is unreactive. The rate constant for the neutral anthrone in benzene-d6 is 2.8 x 10(4) M-1 s-1. Competition experiments with tetramethylethylene in acetonitrile gave a rate constant for reaction alone (kR) of 2.1 x 10(8) M-1 s-1 for the anion.

Anthralin↗
PubMed1989

Evaluation of antithrombogenic thermodilution catheter.

In order to prevent the complications accompanied with pulmonary-artery (Swan-Ganz) catheterization, we have developed an antithrombogenic coating, tradenamed Anthron. Anthron-coated thermodilution catheters show excellent antithrombogenicity due to continuous release of heparin from its surface to the blood stream in animal experiments. As controls, a poly (vinyl chloride) thermodilution catheter was evaluated in the similar manner. All of Anthron-coated thermodilution catheters were completely free from thrombus formation and kept excellent sensing functions for more than 3 days, while in control catheters severe thrombus formations were found both on the surfaces of the catheters and blood vessels. The sensing functions of the control catheters deteriorated with passage of time because of the deposition of the blood constituents on the thermistor.

Animals↗
PubMed1976

A semiautomated micromethod for dextran in the presence of glucose.

A semiautomated method for microanalysis of dextran with anthrone reagent has been developed. The method utilizes the Technicon inulin manifold and other standard AutoAnalyzer equipment. Since the anthrone method is sensitive to plasma glucose, a glocuse oxidase method which eliminates glucose in samples to be analyzed for dextran has also been developed. When solutions containing 50 mg. per 100 ml. of dextran were compared to solutions containing 50 mg. per 100 ml. of dextran plus up to 200 mg. per 100 ml. glucose, no significant difference was observed. The coefficient of the anthrone-glucose oxidase method is 0.039, which is within the error of the AutoAnalyzer itself.

Autoanalysis↗
PubMed2004

[Establishment and evaluation on the method for platelet intracellular trehalose determination].

This study was aimed to establish an applicable, convenient and rapid method for platelet intracellular trehalose determination, so that the technique of trehalose loading could be optimized and used in research on freeze-drying platelets. Protein from loaded-trehalose platelets was deposited by using trichloroacetic acid, trechalose concentration in platelets was determinated by sulfuric-anthrone reaction and was analyzed by high performance liquid chromatography (HPLC). The results showed that when platelets were loaded with 1.7% of trehalose in loading solution, intracellular concentration determinated by sulfuric-anthrone reaction was 0.22% and derterminated by HPLC was 0.2%. The recovery rate of trehalose determinated by sulfuric-anthrone reaction was 100.7% and stability and repeatability of results were good. In conclusion, the method is convenient, rapid, accurate and highly sensitive for determination of platelet intracellar trehalose.

Blood Platelets↗
PubMed1992

[Complications and their management in intraarterial infusion chemotherapy].

Complications and its management were evaluated in intraarterial infusion chemotherapy for 188 patients with advanced carcinoma of the digestive organs from 1975 to Sept. 1991. Subjects were divided into four groups: Group I was 62 patients in whom the tip of the catheter without knots was established in the abdominal aorta via celiac axis, Group II consisted of 72 patients with the tip of the catheter without knots in the common hepatic artery. Group III had 35 patients with the tip of the catheter with knots (Anthron catheter) in the common hepatic artery. Group IV was 19 patients with the tip of the anthron catheter connected to the Infuse A-Port in the common hepatic artery. The most frequent complications seen among Group I, II and III were caused by catheter thrombosis (11.3%) in Group I, spontaneous dislodgement of catheter (26.4%) in Group II and extravasation (20%) in Group III. By using 16 gauge Toray Anthron catheter with Heparin coating on its inner and outer surfaces, the number of complications in Group I and II was kept smaller. Extravasation, on the other hand, has been less frequently seen in Group III by establishing the tip of the catheter at the branching site of the gastroduodenal artery from the common hepatic artery. Complications in Group IV (19 patients) were noted only in 3 patients, i.e., extravasation, subcutaneous necrosis and subcutaneous abscess, respectively. Therefore, we concluded that Group IV showed the most favorable intraarterial infusion chemotherapy with the most infrequent complications.

Aorta↗
PubMed1989

[Glycolipid-patterns of Mycobacterium tuberculosis as an aid for sub-typing].

Total lipids were extracted from Mycobacterium tuberculosis with chloroform-methanol (2:1), applied on a silica-gel thin-layer plate, and developed with chloroform-methanol-acetone (90:10:5). Glycolipids were detected by spraying Anthrone-reagent and heating. Strain H37Rv of M. tuberculosis showed four Anthrone-positive spots, namely trehalose-monomycolate, unidentified glycolipid, trehalose-dimycolate and GL-Rv, and strain H37Ra showed only two spots corresponding to trehalose-monomycolate and trehalose-dimycolate. Other 4 laboratory-stock strains of M. tuberculosis showed glycolipid-pattern identical with either of these two patterns. One hundred and fifty-eight strains of M. tuberculosis, isolated clinically from tuberculosis patients, were classified into 7 types according to their glycolipid-pattern. Twenty-seven strains contained one more Anthrone-positive spot other than those of strain H37Rv. Pattern II was most frequently observed (60 strains), and then pattern I (33 strains), VI (29 strains), IV (13 strains), V (9 strains), VII (8 strains), and III (6 strains). Pattern I corresponded to that of strain H37Ra and pattern VI corresponded to that of strain H37Rv. Glycolipid-pattern did not correlate to clinical features of patients from whom the bacilli had been isolated. A glycolipid, which moved to just under the solvent front, was a new glycolipid which has been found by us and designated as GL-Rv. Chemical structure of GL-Rv was clarified by us as trehalose-polyacyl derivatives (no mycolic acid as the acyl residue). Glycolipid-pattern was very stable and reproducible for each strain of M. tuberculosis.(ABSTRACT TRUNCATED AT 250 WORDS)

Bacterial Typing Techniques↗
PubMed1985

Mechanism of mouse skin tumor promotion by chrysarobin.

The skin tumor-promoting ability of 1,8-dihydroxy-3-methyl-9-anthrone (chrysarobin) was compared with that of 12-O-tetradecanoylphorbol-13-acetate (TPA) and 1,8-dihydroxy-9-anthrone (anthralin) in SENCAR mice. Although dose-response comparisons indicated that chrysarobin was several orders of magnitude less potent than TPA for promoting papilloma formation, this anthrone was 1.5 to 2 times more potent than anthralin. Maximal papilloma responses were achieved by 15 weeks of promotion with TPA whereas at least 25 weeks of promotion were necessary to achieve maximal papilloma responses with chrysarobin or anthralin indicating marked differences in tumor latency between the two classes of compounds. Interestingly, at optimal promoting doses, chrysarobin gave a carcinoma response (22% with 0.3 carcinomas per mouse at 45 weeks) similar to that of TPA suggesting that this compound may be more efficient at promoting carcinomas than papillomas. In two-stage promotion experiments, chrysarobin was incapable of functioning independently as a Stage I or II promoter despite its complete promoting activity. Chrysarobin and TPA were compared at optimal promoting doses for their ability to induce: (a) skin edema, (b) epidermal hyperplasia, and (c) epidermal ornithine decarboxylase. In each case, distinct differences were noted between the two compounds. When taken together, the data support the hypothesis that anthracene-derived skin tumor promoters work at least in part by a mechanism different from the phorbol esters.

Animals↗
PubMed1991

Full replacement of the function of the secondary electron acceptor phylloquinone(= vitamin K1) by non-quinone carbonyl compounds in green plant photosystem I photosynthetic reaction centers.

One-carbonyl quinonoid compounds, fluorenone (fluoren-9-one), anthrone, and their derivatives are introduced into spinach photosystem (PS) I reaction centers in place of the intrinsic secondary electron acceptor phylloquinone (= vitamin K1). Anthrone and 2-nitrofluorenone fully mediated the electron-transfer reaction between the reduced primary electron acceptor chlorophyll A0- and the tertiary electron acceptor iron-sulfur centers. It is concluded that the PS I phylloquinone-binding site has a structure that enables various compounds with different molecular structures to function as the secondary acceptor and that the reactions of incorporated compounds are mainly determined by their redox properties rather than by their molecular structure. Carbonyl groups increase the binding affinity of the quinone/quinonoid compounds but do not seem to be essential to their function. The quinonoid compounds as well as quinones incorporated into the PS I phylloquinone-binding sites are estimated to function at redox potentials more negative than in organic solvents.

Anthracenes↗
PubMed1988

[The effect of exogenous factors on prececal nutrient and amino acid absorption, ascertained from swine with ileo-rectal anastomoses. 3. The effect of crude fiber-rich coarse meal supplements to a basic ration].

In digestion trials using pigs fitted with ileo-rectal anastomoses and parallel intact pigs the influence of wheat straw meal (WSM) or grass meal (GM) supplemented on two different levels to a basal diet was examined with regard to nutrient and amino acid digestibility resp. absorption. Both roughages reduced, obviously because of their high cell wall contents, nearly at equal amounts and partly significantly the precaecal as well as the total digestibility of dry matter, organic matter, carbohydrates (= crude fibre + NFE), crude fibre and hemicelluloses (arabinose, xylose). By the roughage supplementations the crude protein digestibility at the terminal ileum was less reduced than at the end of the total digestive tract, the starch digestibility was hardly influenced and that of ether extract mostly increased. In comparison with precaecal glucose and fructose digestibilities it could be shown that the anthrone method is not suitable for determinations of the precaecal digestibility of water soluble carbohydrates. Beside these compounds other substances in the ileum digesta must evidently be dyed by anthrone too. The crude protein digestibility and the amino acid absorption were precaecally hardly or not reduced by WSM supplementations, therefore it can be concluded that it is possible to dilute the energy concentration in diets (e.g. for sows) by addition of WSM without impairing protein digestion and amino acid absorption. The GM supplementations, however, impaired protein digestibility and amino acid absorption of the whole diet, probably caused by the encrusted, possibly heat damaged protein present in the GM itself. When the roughage supplemented diets were fed, the excretion of nitrogen compounds in the faeces was enlarged due to the more intensive bacterial activity in the hind gut and the additional sorption effects to cell wall substances, so that a too low apparent and also true digestibility is made believe. The applicability of the difference method to the calculation of protein and amino acid digestibilities in roughages is very questionable because of their low contents and the missing additivity. The crude fibre and hemicellulose digestibility values calculated by the difference method demonstrate for the both roughages--in spite of high standard deviations--that the precaecal digestibility of crude fibre is about zero and that of the hard lignified hemicellulose fraction in contrast to the predominantly endospermic hemicelluloses of the basal diet is very low. The faecal crude fibre and hemicellulose digestibilities of WSM--especially on the lower supplementation levels--are markedly worse than those of GM.

Amino Acids↗
PubMed1992

[A modified microanthrone method for the determination of inulin on nanogram scale].

The establishment of a method of determining minute amount of inulin on nanogram scale is essential for applying micropuncture technique to kidney research. In this experiment, we modified a classical microanthrone method which was used by Dirks et al. to determine inulin contained in tubular fluid. A spectrophotometer was installed with a special microcuvette to suit the colorimetry of 10 microliters solution. Linear relation between inulin concentration and absorption was observed in the range of inulin concentration from 2 to 32 nl per 10 microliters anthrone reagent. The accuracy of this microanthrone method was evaluated with inulin recovery test, the mean recovery rate being 101.2 +/- 3.5%. Since tubular fluid did not interfere with the determination of inulin, the tubular fluid which contained inulin could be added directly to anthrone reagent in actual practice. The mean single nephron glomerular filtration rate measured with this modified microanthrone method in 21 rats was 34.9 +/- 1.8 nl/min, which coincided well with the values reported in literature. The results indicate that the measurement of inulin with the above-mentioned microanthrone method is reliable and suitable for the micropuncture study.

Animals↗
PubMed1998

An experimental study on the detection of fructose in Aedes aegypti.

Male and female Aedes aegypti fed a 10% sucrose solution and/or blood were tested to determine the duration of fructose detection in their bodies and the volume of sugar solution they ingested. The limit of detection of fructose by the cold anthrone test was investigated in a series of experiments. Results were applied to the interpretation of sugar feeding by Ae. aegypti collected inside houses in Puerto Rico during times of low (cool season) and high (hot season) dengue transmission in 1996. We conclude that, under our experimental conditions, the cold anthrone test can detect a 10% sucrose solution in male and female Ae. aegypti up to approximately 4 days after ingestion, even the smallest volumes of 10% sucrose solution ingested by experimental mosquitoes are detectable, the test is sensitive enough to detect 0.6 microgram of fructose, and the cutoff point for defining positive fructose values in field-collected females should be based on blood-engorged specimens. We confirmed that female Ae. aegypti collected from natural resting sites inside houses in Puerto Rico seldom, compared to males, contain detectable amounts of fructose.

Aedes↗
PubMed1991

Structure-function relationship of new anthralin derivatives assayed for growth inhibition and cytotoxicity in human keratinocyte cultures.

HaCaT keratinocyte cultures were exposed to twelve hydrophilic anthralin derivatives 1 to 12 with substituents at C-1 and C-8 of the anthrone skeleton, of one H at C-10 and of both H's at C-10 by lacton rings. After 3 microM treatment growth was determined by cellular protein content, 3H-thymidine- and 14C-amino-acid-uptake and cytotoxicity by the release of cytoplasmic LDH into the culture medium. In comparison to acetone control (100%) anthralin suppressed mean protein content, as well as DNA- and protein-synthesis to 33, 28, and 21%, respectively, and the drug revealed an enzyme release of 660%. In relation to the parent drug we found similar cell growth inhibitory effects of compounds 4, 6, 8, 9, 10, and 12. Deriv. 4, 8, and 10 were, however, to some extent less cytotoxic than anthralin, whereas deriv. 6, 9, and 12 were in the same range. An extreme suppression of growth parameters which differed from the anthralin effect by a factor 0.5-0.8 was caused by deriv. 11, showing the same cytotoxicity. Deriv. 1, 2, 3, 5, and 7 did not demonstrate any cytotoxicity. Concerning growth parameters, deriv. 2 induced a slight stimulation, deriv. 3 and 7 were completely ineffective, deriv. 1 and 5 induced slightly to moderately inhibited proliferation but both being much less effective than anthralin. These data indicate that the "minimum structure" concept by Krebs and Schaltegger--claiming 1-hydroxy-9-anthrone as a precondition for clinical antipsoriatic potency--is not valid at least in cell-biological tests and point toward possible usefulness of some experimental model compounds as alternative antipsoriatics.

Anthralin↗
PubMed1998

Antioxidants attenuate anthralin-induced skin inflammation in BALB/c mice: role of specific proinflammatory cytokines.

Anthralin is the most common therapeutic agent among a small number of pro-oxidant, 9-anthrones effective in the topical treatment of psoriasis. However, the usefulness of this drug is diminished by toxic side effects, including skin irritation and inflammation. The activities of anthralin are believed to be mediated by the generation of reactive oxygen intermediates and anthrone radicals produced in the skin. In this study, the dermal inflammatory response to anthralin was determined using a mouse ear swelling test. Maximum ear swelling induced by anthralin coincided with the elevation of cytokine mRNA expression in the skin, including interleukin-6, granulocyte-macrophage colony-stimulating factor, macrophage inflammatory protein-2, and tumor necrosis factor alpha at 24 h post challenge. The role of free radical generation in ear swelling and cytokine modulation were examined by systemic administration of cell permeable and impermeable antioxidants before anthralin challenge. Superoxide dismutase and alpha-tocopherol acetate, but not the glutathione precursor N-acetyl cysteine, were effective inhibitors of anthralin-induced ear swelling and cytokine elevation. Maximum inflammatory cell infiltration occurred 72-96 h post anthralin challenge and was also reduced by antioxidants. These data suggest that oxidative stress, generated at the site of anthralin treatment, alters the expression of dermal chemokines and other cytokines resulting in the recruitment of inflammatory cells. Systemic antioxidant administration may provide opportunities for therapeutic intervention against anthralin-associated toxicities.

Acetylcysteine↗
PubMedDate not supplied

The phytochemical profile and identification of main phenolic compounds from the leaf exudate of Aloe secundiflora by high-performance liquid chromatography-mass spectroscopy.

The phytochemical profile of Aloe secundiflora (Aloeaceae) and the identity of eight major compounds, including the two main constituents, have been determined from the leaf exudate of this ethnoveterinary used species from Kenya and Tanzania. Analytical HPLC-MS studies of the exudate have revealed that it comprises a mixture of phenolic compounds, mainly anthrones (aloenin, aloenin B, isobarbaloin, barbaloin and other aloin derivatives), chromones and phenylpyrones with a low content of polysaccharides and aliphatic compounds. The high percentage of anthrones in the exudate could provide a first line of evidence for the use of the plant in ethnoveterinary practices.

Aloe↗