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At least 271 records · Page 15Linked to original sources

Evidence for autocrine/paracrine growth stimulation by transforming growth factor-alpha during the process of skin tumor promotion.

A single topical application of 12-O-tetradecanoylphorbol-13-acetate (TPA) to mouse skin decreased 125I-labeled epidermal growth factor (EGF) binding in epidermal membrane preparations within 1 h while 1,8-dihydroxy-3-methyl-9-anthrone (chrysarobin) gradually reduced binding with maximum inhibition at 15 h. Subsequently, 125I-EGF binding increased to approximately 200% of control in epidermal membrane preparations from both TPA- and chrysarobin-treated mice. A single application of TPA but not chrysarobin resulted in a rapid translocation of protein kinase C (PKC) to the membrane; however, treatment with both promoters ultimately led to a time-dependent loss of PKC activity in both membrane and cytosol fractions. The initial inhibition of 125I-EGF binding was sustained for at least 24 h after single and multiple treatments with both promoting agents. Acid washing restored EGF binding to control levels in membrane preparations obtained 24 h after a single application, whereas acid washing of membrane preparations obtained 24 h after a second application of TPA or chrysarobin increased binding (2.5-fold and 1.5-fold that of the control, respectively). The presence of increased amounts of ligands for the EGF receptor in tumor promoter-treated epidermis was initially confirmed in 125I-EGF binding competition experiments using NRK-49F cells. A single topical application of TPA or chrysarobin induced elevated levels of transforming growth factor-alpha (TGF-alpha) mRNA at 6 h or 15-24 h, respectively. Elevated levels of a TGF-alpha precursor (21 kDa) were subsequently observed in cytosol and membrane preparations after single and multiple applications of TPA or chrysarobin. These results suggest that repeated topical application of tumor promoters may lead to sustained loss of a negative-feedback mechanism involving phosphorylation at Thr-654 of the EGF receptor by PKC. The concomitant elevation of ligands, such as TGF-alpha, may provide a mechanism for sustained cell proliferation essential for skin tumor promotion.

Animals↗

Microsporidian intrasporal sugars and their role in germination.

The hypothesis that spores of terrestrial and aquatic microsporidia differ in their utilization of sugars was tested by evaluating the sugars in germinated and ungerminated spores of several species in each category. The aquatic species tested were Vavraia culicis, Edhazardia aedis, and Nosema algerae and the terrestrial species were Vairimorpha necatrix, Nosema disstriae, Nosema apis, Vairimorpha lymantriae, and Nosema spp. from Spodoptera exigua and Plutella xylostella. The percentage germination varied between species, ranging between 40 and 92%. Total sugars (anthrone reactive) and reducing sugars (Nelson's test) remained unchanged through germination in the three terrestrial species tested; however, reducing sugars increased significantly in the aquatic species. High-performance liquid chromatography and gas chromatography revealed a preponderance of trehalose in all species and large quantities of sorbitol in all species except N. algerae and E. aedis. Other sugars were present in some species in much lower concentrations. After germination no changes in sugar content were observed in terrestrial species; however, all aquatic species lost trehalose with a concomitant increase in fructose and/or glucose concentrations. Increased osmotic potential from breakdown of trehalose has been postulated to induce germination of the aquatic species, but another explanation must be found for the terrestrial species.

Animals↗

A purified saponin acts as an adjuvant for a T-independent antigen.

Three strains of mice were injected with a T-independent antigen, Escherichia coli 055:B5 polysaccharide (PS) combined with purified saponin, QS-21, isolated from Quillaja saponaria bank. PS was prepared by hydrolysis of lipopolysaccharide (LPS). Nine week old mice were injected intradermally with 60 micrograms PS, as determined by an anthrone assay, with or without 15 micrograms QS-21 on days 0 and 14. On day 22 sera were assayed by EIA for PS specific antibodies. Titers were 11-fold higher in CD-1 mice with QS-21. C3H/HeJ (Ipsd) and C3H/HeSnJ (Ipsr) mice also showed an adjuvant associated increase in titer with saponin. Therefore, LPS responsiveness was not required for the adjuvant effect. PS vaccinated C3H and CD-1 mice with and without QS-21 had similar antibody isotype profiles. IgG2b titers accounted for more than half of the total Ig response. IgG2a was next highest followed by IgG3, IgM, IgG1, and IgA. In comparison, CD-1 mice injected with 0.1 microgram intact LPS had a different LPS specific isotype profile. IgG3 was the highest followed by IgG1, IgG2b, IgM, IgG2a, and IgA.

Adjuvants, Immunologic↗

Quality control of 99mTc-labeled dextran for lymphoscintigraphy.

99mTc-labeled dextran has been suggested as a lymphoscintigraphic agent. However, quality-control results from previous studies have been controversial. In this study, the optimal concentration of stannous ion and pH value were determined to obtain maximal labeling. Paper and thin-layer chromatography showed total labeling efficiency as high as 98.4%. Anthrone test of the supernatant of the segments from thin-layer chromatographic strip was performed. Colorimetric determinations verified that dextran was found in the same locations as the peak radioactivity.

Chromatography, Paper↗

The ageing process in infective larvae of the roundworms Haemonchus contortus and Nippostrongylus brasiliensis: carbohydrate content.

Trehalose, glycogen and traces of glucose were the only anthrone sensitive carbohydrates identifiable. The variation of 13-22% of the dry weight in different populations of Haemonchus contortus L3 is shown to be due to their capacity to synthesize glycogen during ageing. This contrasts with the larvae of Nippostrongylus brasiliensis in which a low level of carbohydrate remains constant with time. It is suggested that H. contortus can convert lipid to carbohydrate as is the case for several other nematodes. H. contortus L3 show some evidence of physiological adaptation.

Aging↗

Metabolic products of microorganisms. 185. The anthraquinones of the Aspergillus glaucus group. I. Occurrence, isolation, identification and antimicrobial activity.

The occurrence of emodin, erythroglaucin, physcion, physcion-9-anthrone, questin, catenarin, and catenarin-8-methyl ether in different species of the Aspergillus glaucus group (genus Eurotium) was investigated. So far catenarin-8-methyl ether (1, 4, 6-trihydroxy-8-methoxy-3-methylanthraquinone) has not been described as a natural product; it was therefore given the name rubrocristin. The chemical and physical properties of rubrocristin are reported. In addition a new violet pigment (C16H12O5) was isolated and characterized by its MS-, IR- and UV-spectra. The antimicrobial properties of all substances were examined in the agar diffusion assay. Gram-positive bacteria were the most sensitive organisms and catenarin was the most active naturally occurring substance. Synthetically obtained 1, 4, 6, 8-tetrahydroxyanthraquinone was slightly more active than catenarin, whereas rubrocristin showed no antibacterial activity.

Anthraquinones↗

Multifunctional analysis of the interaction of anthralin and its metabolites anthraquinone and anthralin dimer with the inner mitochondrial membrane.

We studied the interaction of the antipsoriatic compound anthralin (1.8-dihydroxy-9-anthrone), and its metabolites anthraquinone (1.8-dihydroxy-9.10-anthraquinone) and anthralin dimer (1.8.1'.8'.-tetrahydroxy-10.10'-bis-9[10]-dianthrone) with the inner mitochondrial membrane. Mitochondrial membrane functions such as ubiquinone redox equilibria, redox status of iron sulfur clusters, cyanide-sensitive and cyanide-insensitive oxygen consumption, adenosine triphosphate (ATP) synthesis, ATP hydrolysis, and adenine nucleotide content of mitochondria were analyzed. Anthralin is an inhibitor of mitochondrial oxygen uptake in the presence of ADP and substrate (cyanide-sensitive respiration), inhibits ATP synthesis without affecting ATP hydrolysis, and depletes mitochondria of ATP. Anthralin dimer is a much weaker inhibitor of mitochondrial functions and anthraquinone is almost inactive. Anthralin, but not anthraquinone and anthralin dimer, reverses uncoupler stimulated oxygen consumption, stimulates cyanide-insensitive respiration, reduces mitochondrial ubiquinone-9 and -10 to the corresponding ubiquinols and reduces mitochondrial iron sulfur clusters. Anthralin may induce formation of reactive oxygen species by enhancing autoxidation of mitochondrial components and/or by catalyzed oxidation of anthralin. Taken together, anthralin acts as an electron donor to inner mitochondrial membrane associated redox components, inhibits the electron transport chain, and has an oligomycin-like effect. Anthralin dimer and anthraquinone do not function as electron donors and act by a different reaction mechanism. Respiratory measurements in human keratinocytes revealed similar results as obtained with isolated mitochondria. We suggest that modulation of membrane redox status may be a common concept of anthralin action in target cells such as keratinocytes and neutrophils.

Adenine Nucleotides↗

The pharmacokinetics of subcutaneous dihydroergotamine with and without a dextran 70 infusion.

The pharmacokinetics of subcutaneous dihydroergotamine (DHE) with or without dextran 70 infusion was evaluated in a single- and multiple-dose study in 30 patients. Radioimmunoassay was used to measure plasma DHE and the anthrone method to determine the dextran concentration. In the single-dose study no significant interaction between DHE and dextran was noted with respect to their plasma levels. The absorption of s.c. DHE was rapid and the disappearance curve followed a biphasic pattern, t0.5 alpha being 1.4 and 2.0 h, t0.5 beta 22 and 21 h for DHE and DHE/dextran 70, respectively. In the multi-dose study the trough level of DHE initially had a tendency to rise, in accordance with simulated plasma concentration curves. DHE trough levels were about 0.5 ng/ml and were well above the assumed minimum effective value to induce venoconstriction (0.06 ng/ml). Dextran concentrations were significantly higher when DHE was co-administered, possibly, due to changes in plasma volume. It is concluded that DHE 0.5 mg s.c. twice daily will give an adequate plasma concentration and that there was no important interaction between it and infused dextran 70.

Adult↗

EDTA soluble protein of human mature normal enamel.

Pure human mature enamel was prepared using a careful microdissection technique. After EDTA dissolution, the soluble proteins were recovered representing a concentration of 0.035% in the initial enamel. When the samples were analyzed with polyacrylamide gel electrophoresis, Coomassie Brilliant Blue staining revealed only one sharp fast migrating band, whereas o-toluidine blue, methylene blue, Amido Black 10B, and pyronine red G showed a thin double band at the same migration distance. Ultracentrifugation studies suggested that the proteins were of low molecular weight or of weak density. Absorption spectra showed a strong absorbance at 260 nm. After hydrolysis, amino acid analyses yielded a composition of 25% Gly, 13.5% Glu, 11% Ser, and 11% Pro. Cysteine measured as cysteic acid was present at 2%, and 2% hydroxyproline was found. A carbohydrate content of 15% was estimated by the anthrone method. Glucose, galactose, mannose, and fucose, identified through gas chromatography, were in a molar ratio of 9:4:3:1. Thus the organic matrix of adult human enamel consists of one or possibly two acidic glycoproteins.

Amino Acids↗

Isolation and characterization of a novel polychlorinated biphenyl-degrading bacterium, Paenibacillus sp. KBC101.

The biphenyl-utilizing bacterial strain KBC101 has been newly isolated from soil. Biphenyl-grown cells of KBC101 efficiently degraded di- to nonachlorobiphenyls. The isolate was identified as Paenibacillus sp. with respect to its 16S rDNA sequence and fatty acid profiles, as well as various biological and physiological characteristics. In the case of highly chlorinated biphenyl (polychlorinated biphenyl; PCB) congeners, the degradation activities of this strain were superior to those of the previously reported strong PCB degrader, Rhodococcus sp. RHA1. Recalcitrant coplanar PCBs, such as 3,4,3',4'-CB, were also efficiently degraded by strain KBC101 cells. This is the first report of a representative of the genus Paenibacillus capable of degrading PCBs. In addition to growth on biphenyl, strain KBC101 could grow on dibenzofuran, xanthene, benzophenone, anthrone, phenanthrene, naphthalene, fluorene, fluoranthene, and chrysene as sole sources of carbon and energy. Paenibacillus sp. strain KBC101 presented heterogeneous degradation profiles toward various aromatic compounds.

Bacillaceae↗

Skim soy protein enhances GFR as much as beefsteak protein in healthy human subjects.

BACKGROUND: It has been reported that an acute load of beefsteak (200 g) significantly enhanced the glomerular filtration rate (GFR; inulin clearance and creatinine clearance) in healthy human subjects compared to that in the same subjects in the fasted state. However, no comparative study of the effects of the same amount of vegetable protein on GFR has been reported to date. METHODS: We attempted to compare changes in the GFR (inulin clearance and creatinine clearance) in six healthy male subjects following consumption of the same amount of beefsteak or baked skim soy with soy sauce (protein, 86.9 g) after fasting. The clearance study was performed by conventional methods. Inulin was measured by the anthrone method. Creatinine was measured by the Jaffe rate assay method. Amino-acid analysis of the beefsteak and baked skim soy with soy sauce was done by acid or hydroxide hydrolysis and an amino-acid analyzer. RESULTS: A significant enhancement of the GFR (both inulin clearance and creatinine clearance) was observed following acute loading with beefsteak or baked skim soy with soy sauce, compared to the GFR in the fasted state. No significant difference was observed between the results with beefsteak and the results with baked skim soy with soy sauce. Amino-acid analysis revealed that the total amount of three amino acids (glycine, alanine, and arginine; or serine, alanine, and proline) was almost identical in beefsteak (animal protein) and baked skim soy with soy sauce (vegetable protein). CONCLUSIONS: The present study demonstrated that vegetable protein with the same amino-acid composition could enhance the GFR in healthy subjects as much as animal protein.

Adult↗

Effects of protein-bound polysaccharide isolated from pumpkin on insulin in diabetic rats.

To investigate hypoglycemic substances from pumpkin, protein-bound polysaccharide was isolated by activity-guided isolation from water soluble substances of the fruits. The protein-bound polysaccharide from pumpkin fruits (PBPP) was identified to consist mainly of polysaccharide (approximately 41.21%) and protein (approximately 10.13%) by anthrone test and Lowry-Folin test. Different doses of PBPP were evaluated for hypoglycemic activity and the effect on serum insulin levels in alloxan diabetic rats. The results indicated that PBPP can obviously increase the levels of serum insulin, reduce the blood glucose levels and improve tolerance of glucose. The hypoglycemic effect of big dose PBPP group (1000 mg/kg body weight) excelled that of small dose PBPP group (500 mg/kg body weight) and antidiabetic agent group. The results suggest that the hypoglycemic effect of PBPP depends on the dose and PBPP possesses the possibility of being developed from a new antidiabetic agent.

Animals↗

Quenching of carbohydrate reactions by azide.

Sodium azide, at concentrations that prevent development of microorganisms, interferes with the anthrone and phenol sulfuric acid reactions for carbohydrates, and with the glucose oxidase reaction. It does not affect copper reduction and the ortho-toluidine reaction.

Azides↗

Synthesis and characterization of 6-O-(N-heptylcarbamoyl)-methyl-alpha-D-glucopyranoside, a new surfactant for membrane studies.

A new surfactant, 6-O-(N-heptylcarbamoyl)-methyl-alpha-D-glucopyranoside (HECAMEG, molar mass 335.38 g), was synthesized by a simple and low cost procedure from methyl-alpha-D-glucopyranoside. This surfactant is characterized by a high solubility in water (even at 0 degree C), ultraviolet light transparency in the region useful for protein detection, and a high critical micellar concentration (CMC = 19.5 mM), permitting fast elimination by dialysis. Furthermore, the surfactant is colorimetrically titratable by the anthrone technique and its weak interference in protein titration by the Lowry et al. procedure and the bicinchoninic method is easy to overcome. Two membrane proteins (NADH oxidase and succinate dehydrogenase) and a soluble enzyme (lactoperoxidase) retained full activity in the presence of HECAMEG below or above its CMC. The partial inhibition of beta-lactamase (soluble form) by HECAMEG above the CMC was probably only apparent and due to an interference of the surfactant with the substrate rather than a direct effect on the enzyme. HECAMEG was capable of extracting up to 75% of bacteriorhodopsin from the purple membrane of Halobacterium halobium in a nondenatured form as indicated by the spectral properties of the protein. It also solubilized spiralin from the Spiroplasma melliferum membrane with a great selectivity and efficiency, without detectable loss of antigenic properties. These data show that HECAMEG is a very mild surfactant, useful for membrane protein studies.

Bacteria↗

Constituents of salivary supernatant responsible for stimulation of oxygen uptake by the bacteria in human salivary sediment.

The 10,000 g supernatant of wax-stimulated whole saliva was fractionated by gel filtration and its components were tested along with amino acids, small peptides and urea for their ability to stimulate this oxygen uptake, and for their effects on pH. Fractions containing the larger components, the proteins and large peptides, stimulated much less oxygen uptake than unfractionated supernatant, and caused a small decrease in pH. Analysis with anthrone indicated that both these effects were due mainly to the carbohydrate associated with these constituents. In contrast, fractions containing the remaining lower molecular-weight components stimulated substantial oxygen uptake and a rise in pH; both effects were like those seen with whole saliva supernatant. The oxygen effects were attributed mainly to certain amino acids and small peptides in the small molecular-weight fractions. Ornithine, arginine, proline and glutamic acid consistently stimulated oxygen uptake by the oral microflora in a test of 23 amino acids with the sediments of 13 subjects. Ornithine and arginine at the same time stimulated a significant rise in pH, whereas the other two amino acids showed no such effect. Variable and sometimes significant oxygen uptake was seen with alanine, aspartic acid, asparagine, glutamine and cysteine in 4-7 of the subjects; infrequent or no effects were seen with the remainder of the amino acids tested. There was some evidence to suggest that amino acid stimulation of oxygen uptake may be inducible. Urea had no effect on uptake but did contribute significantly to the pH rise. Small peptides containing those amino acids that could stimulate oxygen uptake also stimulated such uptake; peptides without such acids did not.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acids↗

Modification of DNA bases by anthralin and related compounds.

Modification of bases in calf thymus DNA by treatment with the antipsoriatic drug anthralin was studied. The products of DNA bases were identified and their yields measured by gas chromatography-mass spectrometry with selected ion monitoring. Treatment of calf thymus DNA with anthralin significantly enhanced the amount of modified bases above control levels. Purine bases were modified to products identical with those known to be typical of DNA damage induced by hydroxyl radicals. The yields of Fapy-adenine, 8-hydroxyadenine, Fapy-guanine, and 8-hydroxyguanine were maximally increased at an anthralin concentration of 75 microM. A variety of structural analogues of anthralin were also tested at 75 microM were either weaker or stronger hydroxylating agents. It is likely that damage to DNA bases induced by anthrones contributes to their antiproliferative activity. The pharmacological implications of these characteristics of the action of anthralin on DNA bases are discussed.

Adenine↗

A simple spectrophotometric method for quantitative fecal carbohydrate measurement.

A simple quantitative assay was developed for measuring total fecal carbohydrate (CHO) excretion using stools obtained during balance studies performed for fecal fat determination. This spectrophotometric method utilizes Dreywood's anthrone reagent which reacts with equal weights of CHO whether monosaccharide or polysaccharide. Analysis of known dietary CHO solutions yielded greater than 90% of the known theoretical concentration. Recovery of CHO (Polycose) added to fresh stool was greater than 95%, inter-assay coefficient of variation (CV) 6.2%. Stool specimens stored frozen and analyzed over a 21-month period yielded stable results. Fecal CHO excretion/day determined in 32 normal patients, ages 1 month to 13 years, on diets with varying CHO sources, ranged from 0.04 to 0.85 g, average 0.33 g, SD +/- 0.24. Three patients with diseases known to be associated with CHO malabsorption studied showed markedly increased total fecal CHO excretion, up to 53% of their CHO intake. Quantitative fecal CHO excretion in these patients allowed for assessment of the severity of their disease and provided a means for evaluating the use of different dietary CHO sources in their management.

Adolescent↗

Quantitation of fecal carbohydrate excretion in patients with short bowel syndrome.

We performed inpatient balance studies in 11 patients to evaluate the role of carbohydrate malabsorption in the pathogenesis of the diarrhea seen in short bowel syndrome. Stool weight, total reducing substance as measured by Clinitest, and total fecal carbohydrate as measured by anthrone were determined. Patients had markedly increased fecal carbohydrate excretion, up to 65% of dietary carbohydrate intake. When the diet contained oligosaccharides, measures of total reducing substance greatly underestimated fecal carbohydrate excretion and were unreliable for quantitation. Stool weight correlated with total fecal carbohydrate excretion and with total reducing substance (r = 0.79, p less than 0.001). Multiple balance studies in 2 patients suggested a relationship between both the amount and type of dietary carbohydrate and fecal carbohydrate excretion. These studies suggest that carbohydrate malabsorption is a major cause of the watery diarrhea and subsequent fluid, electrolyte, and acid-base imbalance seen in patients with short bowel syndrome.

Child↗