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Dermatopharmacology of salicylic acid. II. Epidermal antihyperplastic effect of salicylic acid in animals.

The influence of salicylic acid on pathological epithelial proliferation has been evaluated by means of the peidermal hyperplasia inhibition test in the guinea pig. 3% w/w salicylic acid dissolved in ethanol reduced surface epithelial hyperplasia by 15%, i.e. exhibited activity comparable to that of hydrocortisone 0.1% w/w. 1% w/w salicylic acid dissolved in a dimethylacetamide-acetone-ethanol mixture exerted an even greater degree of antihyperplastic activity on the surface epithelium (-18%) and also a marked inhibitory effect on deep epithelial proliferation (-10%).

Acetamides↗

Salicylate intoxication after use of topical salicylic acid ointment by a patient with psoriasis.

The authors describe a therapeutically well managed case of severe salicylate intoxication in a patient with psoriasis who treated himself with 40 percent salicylic ointment applied to approximately 41 percent of his body surface. Nineteen hours after the application of salicylic acid, his blood level of the agent was 6.04 mmol/liter. The patient underwent hemodialysis and was discharged after fourteen days in good general health.

Administration, Cutaneous↗

[Suicidal salicylate intoxications and unintentional percutaneous poisoning with salicylic ointment].

Suicidal salicylate poisonings are presented in 49 patients, 33 women and 16 men, aged from 18 to 71 (mean 37) years. Mixed poisonings with multiple agents were four times more frequent, had more severe clinical course and demanded longer hospitalisation than acute intoxications with salicylates alone. Four patient were over 65 years old (8.2%). Difficult economic situation of geriatric population in Poland had no effect on frequency of suicidal attempts in the studied cohort. There were no fatalities in the studied group. There were two unintentional systemic poisonings due to topical administration of the 10% salicylic acid ointment for wide spread skin lesions (more than 80% of body surface) in two patients with exudative psoriasis.

Administration, Cutaneous↗

[Salicylic acid and urea--possible modification of the keratolytic effect of salicylic acid by urea].

The keratolytic effect of salicylic acid was measured with a colorimeter after staining the outer layers of the stratum corneum with silver nitrate. Salicylic acid was used in a concentration of 5% or 10% in vaseline or of 5% in an improved vehicle. The results of two experiments showed that improvement of the vehicle increased the keratolytic effect of salicylic acid and that urea played a major role in this improvement.

Dermatologic Agents↗

Equilibrium studies on chromium(III) complexes of salicylic acid and salicylic acid derivatives in aqueous solution.

The complexes of chromium(III) ion formed by salicylic acid, SA(H(2)L), and its derivatives (H(2)L): 5-nitrosalicylic acid (5-NSA), 5-sulphosalicylic acid (5-SSA) were investigated by means of potentiometry and spectroscopy, at 25 degrees C and in ionic strength of 0.1 M KNO(3) and 0.1 M KCl, respectively. Over the acidic pH range, the coordination of Cr(III) ion to SA and its derivatives in 1 : 1 mole ratio occurs, CrL(+) type complex is formed. In the excess of ligand, the coordination of the second ligand molecule is somewhat hindered; as a result CrL(HL) type complex occurs. Their existences were verified and their formation constants were determined. At near neutral pH, CrL(OH) and CrL(HL)(OH)(-) type hydroxo complexes formed by hydrolytic equilibria and their formation constants were also defined. The stabilities of Cr(III) complexes of SA and its derivatives decrease in the following order: SA>5-SSA>5-NSA. The formation constants of Cr(III) complexes of SA and its derivatives are in comparable ranges with the corresponding complexes of the 2,x-dihydroxybenzoic acid (2,x-DHBA) of Cr(III) ion. The stabilities of SA complexes for V(IV), Cr(III) and Fe(III) ions that have similar ionic radii, increase in the order VOL<CrL(+)<FeL(+). It is worthwhile noting that the binding ability of Cr(III) to salicylate ion is four orders of magnitude higher than to transferin, which is a blood serum protein and is associated with insulin action.

Chromium↗

Hepatic and cerebral pathology findings in children with fatal salicylate intoxication: further evidence for a causal relation between salicylate and Reye's syndrome.

Histology or necropsy records of 13 children with accidental or therapeutically induced salicylate intoxication were examined for the presence of hepatic and cerebral pathology findings characteristic of Reye's syndrome. Liver sections stained with haematoxylin and eosin showed intrahepatocytic microvesiculation (10 of 12 children) and absence of significant inflammation or necrosis (10 of 12 children). All 6 specimens of liver tissue stained with oil red O showed intrahepatocytic microvesicular fat with central hepatocytic nuclei distributed either diffusely throughout the lobule or more prominently in the lobular periphery. Liver tissue stained with the periodic-acid/Schiff method showed complete absence of stainable glycogen in 5 of 6 children. 9 of 12 children for whom information was available had cerebral oedema. It is concluded that the light-microscopy hepatic findings and the gross cerebral findings for the majority of these children with salicylate intoxication are the same as those for children with Reye's syndrome.

Accidents, Home↗

Manipulation of salicylate content in Arabidopsis thaliana by the expression of an engineered bacterial salicylate synthase.

Salicylic acid (SA) plays a central role as a signalling molecule involved in plant defense against microbial attack. Genetic manipulation of SA biosynthesis may therefore help to generate plants that are more disease-resistant. By fusing the two bacterial genes pchA and pchB from Pseudomonas aeruginosa, which encode isochorismate synthase and isochorismate pyruvate-lyase, respectively, we have engineered a novel hybrid enzyme with salicylate synthase (SAS) activity. The pchB-A fusion was expressed in Arabidopsis thaliana under the control of the constitutive cauliflower mosaic virus (CaMV) 35S promoter, with targeting of the gene product either to the cytosol (c-SAS plants) or to the chloroplast (p-SAS plants). In p-SAS plants, the amount of free and conjugated SA was increased more than 20-fold above wild type (WT) level, indicating that SAS is functional in Arabidopsis. P-SAS plants showed a strongly dwarfed phenotype and produced very few seeds. Dwarfism could be caused by the high SA levels per se or, perhaps more likely, by a depletion of the chorismate or isochorismate pools of the chloroplast. Targeting of SAS to the cytosol caused a slight increase in free SA and a significant threefold increase in conjugated SA, probably reflecting limited chorismate availability in this compartment. Although this modest increase in total SA content did not strongly induce the resistance marker PR-1, it resulted nevertheless in enhanced disease resistance towards a virulent isolate of Peronospora parasitica. Increased resistance of c-SAS lines was paralleled with reduced seed production. Taken together, these results illustrate that SAS is a potent tool for the manipulation of SA levels in plants.

Amino Acid Sequence↗

Influence of renal failure in rats on the disposition of salicyl acyl glucuronide and covalent binding of salicylate to plasma proteins.

Many acyl glucuronides are labile, reactive and form covalent adducts with proteins. In the present experiment, the stability of salicyl acyl glucuronide (SAG), its reactivity with serum albumin in vitro and the influence of renal failure in rats on the disposition of SAG and covalent binding of salicylate (SA) to rat plasma proteins were investigated. In vitro studies showed that SAG was hydrolyzed to SA or undergoes isomerization to positional isomers. The half-life of SAG was 1.3 hr in 0.15 M phosphate buffer at pH 7.4 and 37 degrees C, but the stability of its isomers was much greater with an apparent half-life of 19 hr. Incubation of SAG in solutions of human serum albumin revealed the formation of covalent adducts with the protein, with maximal binding of 2.8% of total SA equivalents added to the solution. For in vivo studies, one group of rats was administered 5 mg/kg of uranyl nitrate i.p. to induce renal failure. After administration of 100 mg/kg of SA i.v., the AUC 0-26 hr of SAG in rats with renal failure was 9 times higher than that observed in control rats. The apparent clearance of SA decreased from 64 +/- 21 ml/hr in control rats 28 +/- 8.7 ml/hr in rats with renal failure. The level of SA covalent adducts with plasma proteins reached about 0.5 ng/mg of protein in control rats, whereas in rats with renal failure the binding was increased significantly and achieved an average peak concentration of 18 ng/mg of protein when measured 26 hr after dosing. The data indicate that reactive SAG can accumulate in renal failure which then increases covalent addition of SA to proteins. Such binding may have a role in enhancing the potential for toxicity of acidic drugs such as SA in renal disease.

Animals↗

Dermatopharmacology of salicylic acid. III. Topical contra-inflammatory effect of salicylic acid and other drugs in animal experiments.

The acute contra-inflammatory effects of salicylic acid, three standard dermatocorticoids and four contact antiphlogistics have been investigated by means of a UV dermatitis inhibition test in the guinea pig. The substances tested had a distinct inhibitory effect on the development of erythema and can be ranked in the following ascending order of activity (percent of maximum possible score): bufexamac = 36%, salicylic acid = 37%, hydrocortisone = 44%, acetylsalicylic acid = 48%, flumethasone pivalate = 51%, fluocinolone acetonide = 51%, phenylbutazone = 56%, and indomethacin = 58%.

Administration, Topical↗

Teratogenicity of edoferon kappa A, a molecule derived from salicylate, in cultured rat embryos: differences from salicylate and interaction with free oxygen radical scavenging enzymes.

The effect of edoferon kappa A (E-KA), a non-specific immunomodulatory and anti-neoplastic chemical substance derived from the methyl form of salicylate (acetyl salicylic acid; ASA), on mammalian embryos was studied and compared to the effects of ASA. Rat embryos were cultured in vitro from 9.5 days of gestation for 48 h. E-KA (0.1-12.8 mg/ml) and ASA (0.1-0.6 mg/ml) were added to the whole rat serum. To investigate the interaction of these molecules with antioxidant agents, the lowest effective concentrations of E-KA (0.6 mg/ml) and ASA (0.3 mg/ml) for all parameters were added to the culture media in the presence of superoxide dismutase (SOD) (30 U/ml) or glutathione (0.5 mumol/ml). The growth and development of embryos was compared and each embryo was evaluated for the presence of any malformations. E-KA and ASA decreased growth and development in a concentration-responsive manner. There was also a concentration-related increase in overall dysmorphology (haematoma in the yolk sac and neural system, open neural tube, abnormal tail torsion and the absence of fore limb bud). There were no statistically significant differences between the control and embryos grown in the presence of 0.1-0.4 mg/ml E-KA, although the effects of ASA started at a concentration of 0.2 mg/ml. Embryos showed significant growth retardation in all scoring criteria and severe malformations when 0.5-3.2 mg/ml E-KA and 0.3-0.6 mg/ml ASA were added. When SOD was added, there was a significant decrease in the incidence of malformations and growth and developmental parameters were increased but this decrease never reached the control level. We concluded that E-KA has direct toxic effects on the developing embryo but at much higher concentrations than ASA, and the teratogenic effects of these molecules might be related to free oxygen radicals.

Animals↗

Pharmacokinetic and bacteriological study of cefadroxil-salicylate and josamycin-salicylate drug regimens.

Eye, nose, throat and bronchopulmonary infections are frequently associated with inflammatory symptoms. This often leads the clinician to prescribe a combination of an anti-inflammatory and an antibiotic. Cefadroxil and josamycin are among the antibiotics most frequently used in these infections, and they are often combined with acetylsalicylic acid in various pharmaceutical formulations. The study of possible pharmacokinetic and bacteriological interactions was performed in healthy volunteers who received in a crossover protocol each of the two antibiotics, either alone or combined with acetylsalicylic acid or lysine acetylsalicylate. No marked pharmacokinetic interaction was noted except an increase in the AUC for plasma concentrations of cefadroxil when combined with a salicylate. A greater uniformity of kinetic profiles was seen with cefadroxil than with josamycin. Lastly, with the exception of one strain, the salicylates did not alter the antibacterial activity of cefadroxil.

Adult↗

Influence of salicylic acid on H2O2 production, oxidative stress, and H2O2-metabolizing enzymes. Salicylic acid-mediated oxidative damage requires H2O2.

We investigated how salicylic acid (SA) enhances H2O2 and the relative significance of SA-enhanced H2O2 in Arabidopsis thaliana. SA treatments enhanced H2O2 production, lipid peroxidation, and oxidative damage to proteins, and resulted in the formation of chlorophyll and carotene isomers. SA-enhanced H2O2 levels were related to increased activities of Cu,Zn-superoxide dismutase and were independent of changes in catalase and ascorbate peroxidase activities. Prolonging SA treatments inactivated catalase and ascorbate peroxidase and resulted in phytotoxic symptoms, suggesting that inactivation of H2O2-degrading enzymes serves as an indicator of hypersensitive cell death. Treatment of leaves with H2O2 alone failed to invoke SA-mediated events. Although leaves treated with H2O2 accumulated in vivo H2O2 by 2-fold compared with leaves treated with SA, the damage to membranes and proteins was significantly less, indicating that SA can cause greater damage than H2O2. However, pretreatment of leaves with dimethylthiourea, a trap for H2O2, reduced SA-induced lipid peroxidation, indicating that SA requires H2O2 to initiate oxidative damage. The relative significance of the interaction among SA, H2O2, and H2O2-metabolizing enzymes with oxidative damage and cell death is discussed.

Arabidopsis↗

Penetration of salicylic acid and salicylate into the multilayer membrane system and into the human horny layer.

Using a multilayer membrane system and human horny layer the difference in the penetration of salicylic acid (SA) and its sodium (Na-S) and choline (Ch-S) salts from topical formulations was studied. It was found Na-S and Ch-S were markedly accumulated in the first membrane of the three layer membrane system used. In contrast, a rapid penetration into all three membranes was observed when SA was used. Similar penetration profiles were obtained in human horny layer. Hence, the use of the salts of SA appears to be more suitable for the application as keratolytic.

Epidermis↗

The salicylate trapping method: is oxidation of salicylic acid solution oxygen and time dependent and metal catalysed?

For a microdialytic trapping method we systematically investigated changes in concentrations of 2,5-dihydroxy-benzoic acid (2,5-DHBA) and 2,3-dihydroxy-benzoic acid (2,3-DHBA) in freshly prepared solutions of salicylic acid (SA). The solvent was 0.9% saline exposed to different atmospheric concentrations of oxygen (0, 21, and 100%). The solutions were treated by freezing-thawing and an ultrasonic bath in presence and absence of aluminium foil. Without aluminium the concentrations of 2,5-DHBA and 2,3-DHBA kept constant over an observed period of 160 min on different levels from below 20 ng/ml to about 100 ng/ml. In presence of aluminium the concentrations increased to maximum 307 ng/ml after 160 min. Ultrasonic irradiation amplified this effect to maximum 341 ng/ml. HPLC/ECD processing and quantitative analysis of dihydroxy-benzoic acids (DHBAs) in microdialysis may be artificially influenced by varying oxygen environment and metal catalysis.

Aluminum↗

Two tobacco genes induced by infection, elicitor and salicylic acid encode glucosyltransferases acting on phenylpropanoids and benzoic acid derivatives, including salicylic acid.

Two tobacco genes (TOGT) with homology to glucosyltransferase genes known to be induced by salicylic acid (SA) also responded rapidly to a fungal elicitor or to an avirulent pathogen. SA, although an efficient inducer, was shown not to be essential in the signal transduction pathway regulating TOGT gene expression during the resistance response. Recombinant TOGT proteins produced in Escherichia coli exhibited low, but significant, glucosyltransferase activity towards SA, but very high activity towards hydroxycoumarins and hydroxycinnamic acids, with glucose esters being the predominant products. These results point to a possible important function in defense of these glucosyltransferases in conjugating aromatic metabolites prior to their transport and cross-linking to the cell wall.

Benzoates↗

Metabolic profiling of oxylipins upon salicylate treatment in barley leaves--preferential induction of the reductase pathway by salicylate(1).

In barley leaves, 13-lipoxygenases (13-LOXs) are induced by salicylate (SA) and jasmonate. Here, we show by metabolic profiling that upon SA treatment, free linolenic acid and linoleic acid accumulate in a 10:1 ratio reflecting their relative occurrence in leaf tissues. Furthermore, 13-LOX-derived products are formed and specifically directed into the reductase branch of the LOX pathway leading mainly to the accumulation of (13S,9Z,11E,15Z)-13-hydroxy-9, 11,15-octadecatrienoic acid (13-HOT). Under these conditions, no accumulation of other products of the LOX pathway has been found. Moreover, exogenously applied 13-HOT led to PR1b expression suggesting for the time a role of hydroxy polyenoic fatty acid derivatives in plant defense reactions.

Enzyme Induction↗

Ozone-induced ethylene production is dependent on salicylic acid, and both salicylic acid and ethylene act in concert to regulate ozone-induced cell death.

Ethylene is known to influence plant defense responses including cell death in response to both biotic and abiotic stress factors. However, whether ethylene acts alone or in conjunction with other signaling pathways is not clearly understood. Ethylene overproducer mutants, eto1 and eto3, produced high levels of ethylene and developed necrotic lesions in response to an acute O3 exposure that does not induce lesions in O3-tolerant wild-type Col-0 plants. Treatment of plants with ethylene inhibitors completely blocked O3-induced ethylene production and partially attenuated O3-induced cell death. Analyses of the responses of molecular markers of specific signaling pathways indicated a relationship between salicylic acid (SA)- and ethylene-signaling pathways and O3 sensitivity. Both eto1 and eto3 plants constitutively accumulated threefold higher levels of total SA and exhibited a rapid increase in free SA and ethylene levels prior to lesion formation in response to O3 exposure. SA pre-treatments increased O3 sensitivity of Col-0, suggesting that constitutive high SA levels prime leaf tissue to exhibit increased magnitude of O3-induced cell death. NahG and npr1 plants compromised in SA signaling failed to produce ethylene in response to O3 and other stress factors suggesting that SA is required for stress-induced ethylene production. Furthermore, NahG expression in the dominant eto3 mutant attenuated ethylene-dependent PR4 expression and rescued the O3-induced HR (hypersensitive response) cell death phenotype exhibited by eto3 plants. Our results suggest that both SA and ethylene act in concert to influence cell death in O3-sensitive genotypes, and that O3-induced ethylene production is dependent on SA.

Arabidopsis↗

High-affinity salicylic acid-binding protein 2 is required for plant innate immunity and has salicylic acid-stimulated lipase activity.

Salicylic acid (SA) is a critical hormone for signaling innate immunity in plants. Here we present the purification and characterization of SA-binding protein 2 (SABP2), a tobacco protein that is present in low abundance and specifically binds SA with high affinity. Sequence analysis predicted that SABP2 is a lipase belonging to the alpha/beta fold hydrolase super family. Confirming this prediction, recombinant SABP2 exhibited lipase activity against several synthetic substrates. Moreover, this lipase activity was stimulated by SA binding and may generate a lipid-derived signal. Silencing of SABP2 expression suppressed local resistance to tobacco mosaic virus, induction of pathogenesis-related 1 (PR-1) gene expression by SA, and development of systemic acquired resistance. Together, these results suggest that SABP2 is an SA receptor that is required for the plant immune response. We further propose that SABP2 belongs to a large class of ligand-stimulated hydrolases involved in stress hormone-mediated signal transduction.

Amino Acid Sequence↗