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PubMed1987

Amino sugars in the glycoprotein toxin from Bacillus thuringiensis subsp. israelensis.

The carbohydrate content of purified Bacillus thuriniensis subsp. israelensis crystal toxin was determined by six biochemical tests, column chromatography on an amino acid analyzer, and the binding of 11 fluorescent lectins. The crystals contained approximately 1.0% neutral sugars and 1.7% amino sugars. The amino sugars consisted of 70% glucosamine and 30% galactosamine. No N-acetylneuraminic acid (sialic acid) was detected. The presence of amino sugars was confirmed by the strong binding of fluorescent wheat germ agglutinin and the weak binding of fluorescent soybean agglutinin. These lectins recognize N-acetyl-D-glucosamine and N-acetyl-D-galactosamine, respectively. The lectin-binding sites appeared evenly distributed among the protein subunits of the crystal. The sugars were covalently attached to the crystal toxin because wheat germ agglutinin still bound alkali-solubilized toxin which had been boiled in sodium dodecyl sulfate, separate by polyacrylamide gel electrophoresis, and transferred to nitrocellulose membranes. This study demonstrates the covalent attachment of amino sugars and indicates that the B. thuringiensis subsp. israelensis protein toxins should be viewed as glycoprotein toxins. The crystals used in the present study were purified on sodium bromide density gradients. Studies employing crystals purified on Renografin density gradients can give artificially high values for the anthrone test for neutral sugars.

Bacillus thuringiensis↗
PubMed1968

Production of a capsular polysaccharide by a marine filamentous fungus.

The spent seawater medium of 4-day-old-cultures of the filamentous marine fungus Leptosphaeria albopunctata had a high viscosity after the fungus was collected by high-speed centrifugation. Microscopic examination of uncentrifuged mycelium suspended in India ink revealed that the viscosity resulted from capsular material. These capsules became disassociated from the mycelium during centrifugation. Precipitation of the medium of centrifuged cultures with 95% ethyl alcohol yielded a highly anthrone-positive polysaccharide material, composed of large amounts of glucose and minute amounts of mannose. Time course studies of the nutritional requirements for capsular polysaccharide production revealed that the capsular material was produced in large amounts, and on a wide variety of sugars, during the period of rapid growth, but was quickly degraded and presumably remetabolized in older cultures. The amount of capsular material produced was enhanced by NaCl concentrations above that of artificial seawater, and KCl could be substituted for NaCl. The salts MgCl(2) and CaCl(2) were also required for capsule production by L. albopunctata, although growth was obtained in cultures without added amounts of these constituents. The possible role of these salts in the metabolism of the fungus is discussed.

Ascomycota↗
PubMed1998

Glycogen assay for diagnosis of female genital Chlamydia trachomatis infection.

Chlamydia trachomatis can synthesize glycogen at various stages in its developmental cycle. The glycogen content of female genital epithelial cells was detected by anthrone, and the results were compared with those from PCR. A total of 320 cervical samples were examined. Of 92 specimens that were positive by PCR, 78 were positive and 14 were negative by the glycogen assay. Of 228 specimens that were negative by PCR, 220 were negative and 8 were positive by the glycogen assay. The sensitivity and specificity of the glycogen assay obtained from these data were 84.8% (78 of 92) and 96.5% (220 of 228), respectively. Use of the glycogen assay to detect the glycogen content in genital epithelial cells may be helpful in the diagnosis of C. trachomatis infection. This is an easy, fast, and inexpensive assay and can be done in less-sophisticated labs.

Cervix Uteri↗
PubMed2003

Isolation and partial characterisation of a new antiproliferative substance from human leucocytes inhibiting growth of Candida albicans.

AIM: To purify and partially characterise a fraction from human leucocytes containing a substance cytotoxic to Candida albicans. METHODS: Leucocytes were isolated from the buffy coats of healthy blood donors. The cytotoxic factor (CF) was isolated from the soluble fraction of the cells. A cell lysate was passed through a filter with a cut off value of 3 kDa, and the filtrate was processed by anionic exchange chromatography and gel filtration. The purified CF was analysed for its chemical and biological properties. The cytotoxicity of CF was tested on C albicans grown on agar plates. RESULTS: Mass spectrometry showed a molecular mass of 2.148 kDa. CF was found in polymorphonuclear neutrophilic cells only. No amino acids were detected, and a low ultraviolet absorbance at 260 nm and resistance to nuclease indicate the absence of nucleic acids. An anthrone test was positive for carbohydrate. The substance was soluble in water. CF showed a dose related cytotoxicity in the range of 0.1-1 mg/ml. The cytotoxic effect was abrogated by zinc ions. Preliminary testing indicated that CF also had cytotoxic effects against some bacteria. CONCLUSIONS: This report describes a factor from isolated human leucocytes that is cytotoxic to C albicans. The substance contains a carbohydrate moiety, whereas no amino acids were detected. The cytotoxicity can be abrogated by zinc ions in vitro. This substance is probably part of the repertoire by which leucocytes prevent infections.

Anions↗
PubMed1984

Preliminary characterization of inhibitors of Neisseria gonorrhoeae produced in vitro by Eubacterium limosum.

Among anaerobic bacteria normally found in the urogenital flora, Eubacterium limosum was found to inhibit the in vitro growth of Neisseria gonorrhoeae. The antigonococcal activity produced by E. limosum was soluble in methanol and in a chloroform--methanol mixture (30:70). The fraction soluble in chloroform--methanol (30:70) yielded eight absorbance peaks when chromatographed on Bio-Gel P-2 and the inhibitory activity was found in the first two peaks. This activity was not absorbed on DEAE Sephacel and was eluted with distilled water in a peak considered as peak 1, on which preliminary characterization was done. The inhibitory activity of peak 1 was found to be heat and pH resistant and not susceptible to proteases, lipase, or amylases. When peak 1 was chromatographed on cellulose paper using a butanol--acetic acid (4:1) solvent system, eight different spots were detected upon spraying the paper with ninhydrin. No spot was detected with anthrone, bromothymol, nor Sudan black reagents used for the detection of carbohydrates and lipids. Based on sodium dodecyl sulfate-urea polyacrylamide gel electrophoresis and gel chromatography on Sephadex G-25, peak 1 appeared either as a diffuse band and as a single peak, respectively. The molecular weight of the inhibitory complex was estimated to be 2400. All these results suggest that the antigonococcal activity produced by E. limosum is composed of more than one low molecular weight amino compound.

Bacteriocins↗
PubMed1991

Isolation, ultrastructure, and partial characterization of collagen from the perineurium of the Florida lobster, Panulirus argus.

Highly concentrated extracellular filaments in the perineurium of the Florida spiny lobster, Panulirus argus, were isolated using ultracentrifugation and linear sucrose gradients. The pellet obtained was highly enriched for the filaments as observed by transmission electron microscopy. Fibril diameter and axial periodicity measurements were obtained from filaments positively and negatively stained with uranyl acetate. A period between 14.0 and 25.0 nm and an average fibril diameter of 15.0 nm were observed. The filaments proved resistant to solubilization by most conventional agents and by several collagenases. NaOH (0.1 M at 100 degrees C) safely dissolved the filaments for measurements of protein content by the Lowry method and carbohydrate content with anthrone reagent. These tests revealed a protein content of approximately 84% and a high carbohydrate content of approximately 15%. Polyacrylamide electrophoresis of an acid-pepsin filament extract revealed a highly concentrated band (approximately 100,000) corresponding to the alpha-1 and alpha-2 bands of vertebrate type I collagen. Wide angle X-ray diffraction yielded meridional reflections that confirmed the filaments as collagen when compared with mammalian collagen X-ray diffraction. The amino acid composition was determined with a computer-assisted Beckman amino acid analyzer, which showed a glycine content of 279 residues/1000. Hydroxylysine and hydroxyproline were present in lower concentrations than expected.

Amino Acids↗
PubMed1988

Effect of sennosides and related compounds on intestinal transit in the rat.

The effect of pure sennosides A + B on large intestinal transit (LIT) was investigated in the rat. LIT was defined as the time from intracecal administration of a color marker through a chronically implanted catheter until first appearance of colored feces. Sennosides (50 mg/kg, administered orally 2-24 h before the marker) reduced LIT from greater than 6 h in controls to a minimum of 30-20 min after a 4- or 6-hour pretreatment. Longer pretreatment times increased LIT again reaching normal values after 24 h. Intracecal administration of sennosides and their natural metabolites (sennidins A + B, rhein-9-anthrone, rhein) simultaneously with the marker accelerated LIT to approximately 50-70 min. The laxative effect was less pronounced after rhein compared with the other compounds. Indometacin, loperamide, and calcium-channel antagonists (verapamil, nifedipine) partially antagonized the effect of intracecal sennosides on LIT and delayed, but did not suppress, appearance of soft feces.

Animals↗
PubMed1988

Metabolism of sennosides--an overview.

The metabolism of sennosides is discussed in view of the results obtained during the last years. Rhein anthrone is to be considered as the ultimate active form produced by microorganisms in the colon. Several contributions of this senna symposium bring complementary information of utmost interest.

Animals↗
PubMed1988

New aspects on the metabolism of the sennosides.

Pure sennoside B was administered to rats. On appearance of the first wet faeces, sennoside B and its metabolites were determined in different parts of the alimentary tract, in faeces and in the urine. The total recovery of unchanged sennoside B and its metabolites was determined by alkali fusion followed by colorimetry and high-pressure liquid chromatography (HPLC). Alkali fusion in 1 N sodium hydroxide solution formed red solutions with sennosides and sennoside derivatives. The molar absorbance of sennosides A and B, sennidin B monoglucoside, sennidins, rhein, danthron, dithranol, rhein-8-glucoside and rhein anthrone at wavelengths of 505-530 nm related approximately to the number of ionizable hydroxy groups in the molecule. Brown polymerized products were isolated from the senna drug. The colour intensity of these products was approximately the same by weight as that of the sennosides themselves, although sennidins could no longer be freed from these by acid hydrolysis. After administration of sennoside B, the average sum of unchanged glucoside and known metabolites in different parts of the gastrointestinal tract and faeces of rats was 61.6% according to HPLC and 92.8% according to the alkali fusion procedure. This difference is indicative of the presence of substances which are no longer identifiable as sennoside derivatives, either by HPLC or by other classical chromatographic methods. Sennosides seem to be partly present in the alimentary tract in polymerized or bound form. The alkali fusion method may be useful in connection with the isolation of as yet unknown metabolites of the sennosides in the gastrointestinal tract.

Alkalies↗
PubMed1980

Glycogen deposition in fetal mouse tissues and the effect of dexamethasone.

Glycogen in fetal mouse tissues was isolated and determined using the anthrone method on gestational days 14-19. Fetal liver showed rapid glycogen deposition after gestational day 17, reaching a value of 46.7 mg/g wet weight on day 19. Heart and placenta showed sizable glycogen stores earlier, with peak values of 21.7 and 12.8 mg/g, respectively, on day 16 which declined thereafter. Lung maintained approximately 10 mg/g days 16-19; gut and brain contained approximately 2 and 4 mg/g, respectively. days 14-19. After injection of dexamethasone into mothers on gestational day 15.5, 16 h prior to assay, the glycogen content of liver increased approximately tenfold, while placental glycogen declined. This indicates that corticosteroids merely accelerate the pattern observed in the normal course of gestation.

Animals↗
PubMed1987

Dithranol: further investigations on the structure-activity-relationship. The new series of the 10-mono-alkyldithranol derivatives.

The in vitro stability of 10-ethyldithranol (1,8-dihydroxy-10-ethyl-9(10H)-anthracenone) and its effects on healthy and psoriatic skin is investigated. 10-Ethyl-dithranol as a representative of the new class of 10-alkyldithranol derivatives neither stains nor irritates and has no antipsoriatic activity at all. This is due to the relatively stable substituent at carbon 10 which enables the fast oxidation processes necessary for the antipsoriatic effect. Therefore, antipsoriatically active dithranol derivatives require a free or easily liberated 10-position and hence the so-called 'minimum structure of antipsoriatic anthrones' keeps its validity.

Anthralin↗
PubMedDate not supplied

Effect of dextran 40 on endothelial binding and vesicle loading of ferritin in rabbit aorta.

The effects of dextran 40 on endothelial binding and vesicle loading of cationized (CF, pl greater than 9.0) and anionic (AF, pl = 3.8 to 4.2) ferritins were investigated in the rabbit aorta. After two minutes of in situ perfusion, both ferritin species entered luminal vesicles and bound to the luminal endothelial membrane with a high surface density at vesicle necks. With CF, the presence of 5% dextran 40 did not change the fraction of vesicles loaded, the number of particles per vesicle, or the average density of particles bound to the luminal membrane, but did reduce the average density of particles adhering to vesicle necks. With AF, dextran 40 increased the percentage of vesicles filled, the average number of particles per loaded vesicle, the membrane particle density, the percentage of loaded vesicle necks, and the average density of particles adhering to vesicle necks. Application of gel chromatography and electrophoresis and determination of carbohydrates in the ferritin and dextran perfusates by the anthrone method after precipitation of the ferritin demonstrated that the dextran and ferritin did not conjugate. We hypothesize that the dextran binds to the endothelial membrane and increases its affinity to AF possibly by shielding negatively charged sites and that it binds in such a way as to offer little steric hindrance to AF particles entering vesicles.

Animals↗
PubMed1988

The relationship between plaque pH, plaque acid anion profiles, and oral carbohydrate retention after ingestion of several 'reference foods' by human subjects.

The primary aim of this study was to rank several reference foods (apple drink, caramel, chocolate, cookie, skimmed milk powder, snack cracker, and wheat flake) according to their plaque pH response as monitored in a panel of 12 volunteers by the plaque-sampling method for comparison with data previously reported with other methods used to assess cariogenicity potential. Secondary experiments (using subsets of the panel of subjects) were undertaken in an attempt to elucidate some of the reasons for the observed plaque pH changes. Oral carbohydrate retention was measured at a single time period after food use as total anthrone-positive carbohydrate material, and as specific acidogenic sugars by gas-liquid chromatography after gel-exclusion chromatography. The concentrations of acid anions in the plaque fluid after food consumption were measured by isotachophoresis eight min after food use. According to the plaque pH response, apple-flavored fruit drink and chocolate were the most acidogenic foods and skimmed milk powder the least acidogenic. There were significant correlations (p less than 0.05) between the plaque pH data and lactate-plus-acetate concentrations in plaque fluid, but the correlations between the pH data and any of the carbohydrate retention parameters were not significant.

Acids↗
PubMed1998

Hypertrehalosaemic neuropeptides decrease levels of the glycolytic signal fructose 2,6-bisphosphate in cockroach fat body

In cockroach fat body, trehalogenesis and glycolysis compete for glucose phosphates as common substrates. During trehalogenesis, glycolysis is inhibited, although the mechanism responsible for this is not known. Incubation of the isolated fat body from the Argentine cockroach Blaptica dubia with an extract of the corpora cardiaca containing as little as 0.005 gland equivalents ml-1 of incubation medium increased the release of trehalose (anthrone-positive material) from the tissue by more than 100 %. The content of the glycolytic activator fructose 2,6-bisphosphate was decreased by up to 50 %. A decapeptide was isolated from the corpora cardiaca of B. dubia and shown to be identical to the naturally occurring Blaberus discoidalis hypertrehalosaemic peptide (Bld HrTH), which was also found in the corpora cardiaca. Synthetic Bld HrTH at 2 nmol l-1 and above increased trehalose production and decreased the content of fructose 2,6-bisphosphate to the same extent as did corpus cardiacum extract. The octapeptides Periplaneta americana cardioacceleratory hormones I and II (Pea CAH-I and Pea CAH-II) also had a significant effect on both parameters. Fructose 2,6-bisphosphate is a potent activator of phosphofructokinase from cockroach fat body if the enzyme is assayed at near-physiological concentrations of substrates and effectors. It is suggested that, because of the decrease in fructose 2,6-bisphosphate levels in the fat body, the activity of the key glycolytic enzyme phosphofructokinase is diminished. This can explain the inhibition of glycolytic flux by hypertrehalosaemic peptides which alters the balance of glucose metabolism in favour of trehalose formation.

Journal Article↗
PubMed1998

Studies of aloe. VI. Cathartic effect of isobarbaloin.

The cathartic effect of isobarbaloin, a stereoisomer of barbaloin (compound principally responsible for the cathartic activity of Aloe), was examined in male rats by oral administration. Individual differences in sensitivity in the laxative activity of isobarbaloin and barbaloin was not found. The cathartic activity (ED50) of isobarbaloin in barbaloin positive rats was 19.2 mg/kg, nearly equal to that of barbaloin (19.5 mg/kg). Also, isobarbaloin administered orally was demonstrated to decompose to aloe-emodin-9-anthrone (active metabolite of barbaloin) as well as to barbaloin. Therefore, it is considered that the mechanism underlying the cathartic effect of isobarbaloin is the same as that of barbaloin.

Administration, Oral↗
PubMed1976

Determination of O-hydroxylysyl-glycosides (OHLG) in urine.

A new method for determination of urinary O-hydroxylysyl-glycosides (OHLG) is described. Human urine was extracted with 2 volumes of ethanol, followed by gel-filtration on a column of Sephadex G-10. The presence of OHLG in the alkaline hydrolyzate of the fraction obtained by gel-filtration was proved by paper chromatography and by liquid chromatography. The alkaline hydrolyzate was subjected to Dowex 50 X4 (H+ form) column chromatography, and OHLG were eluted with 1.5N NH4OH. Content of OHLG in the eluate was determined by the anthrone-H2SO4 method in terms of hexose. Excretion rate of OHLG in urine of adults was shown to be 20 mumoles in terms of hexose per day.

Adult↗
PubMed1991

Assay of keratan sulfate as anion-exchanger bound hexose.

An assay for keratan sulfate in papain digests of human intervertebral disc and other tissues has been developed. The digest is applied to the acetate form of a tertiary amine acrylic anion-exchange resin, the oligosaccharide hexose is removed by washing the resin with 0.2 M sodium acetate buffer pH 5.0, then the keratan sulfate is eluted quantitatively with 1.0 M pyridinium sulfate pH 2.5 and assayed for hexose by the anthrone reaction. The keratan sulfate content of human intervertebral disc tissues ranged from 7 to 78 mumole galactose equivalents/g fresh weight; the root mean square error was 2 mumole/g; 10-25 mg of tissue were required. The separation of oligosaccharides from keratan sulfate was confirmed by gel permeation chromatography, sugar composition, ester sulfate analysis, and nuclear magnetic resonance.

Acetates↗
PubMed2006

Noni juice is not hepatotoxic.

Noni juice (Morinda citrifolia) has been approved for use as a safe food within the European Union, following a review of safety. Since approval, three cases of acute hepatitis in Austrian noni juice consumers have been published, where a causal link is suggested between the liver dysfunction and ingestion of anthraquinones from the plant. Measurements of liver function in a human clinical safety study of TAHITIAN NONI Juice, as well as subacute and subchronic animal toxicity tests revealed no evidence of adverse liver effects at doses many times higher than those reported in the case studies. Additionally, M. citrifolia anthraquinones occur in the fruit in quantities too small to be of any toxicological significance. Further, these do not have chemical structures capable of being reduced to reactive anthrone radicals, which were implicated in previous cases of herbal hepototoxicity. The available data reveals no evidence of liver toxicity.

Acute Disease↗