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Salicylate pollutants (SAs) poses a serious threat to marine ecosystems as emerging contaminants. However, the toxic effects of SAs on marine phytoplankton, as well as the potential mechanisms and their ecological risks linked with them, are remain largely unknown. In this study, we aimed to evaluate the toxic effects of salicylic acid (SA) and its 5-substituted derivatives (5-sSA) on the marine diatom Phaeodactylum tricornutum, as well as the potential molecular mechanism involved in the toxicity. Physiological assays conducted on P. tricornutum revealed significant changes in photosynthetic pigments, chlorophyll fluorescence parameters, and antioxidant enzyme activities. The results showed that exposure of P. tricornutum to SAs caused a significant decline in chlorophyll contents and damage to the photosystem II (PSII) core resulting in the decline of photosynthesis. Although the activities of superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) were enhanced, oxidative damage occurred. Transcriptome analysis showed that a large number of differentially expresses genes (DEGs) were significantly enriched in metabolic pathways such as porphyrin metabolism, terpenoid backbone biosynthesis, and carbon fixation in photosynthetic organisms after SA and 5-BrSA treatments. In addition, key genes in transcriptomic metabolic pathways were further analyzed and validated using weighted correlation network analysis (WGCNA) and real-time fluorescence quantitative PCR (qPCR). Considering the above results, SAs mainly inhibit the processes of photosynthesis by repressing the expression of genes involved in secondary metabolite synthesis and photosynthetic carbon sequestration pathways, thus exerting toxic effects on algal cells. The results of the study will provide key data for understanding the ecological risk and toxicity mechanisms of SA pollutants., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. We wish to confirm that there are no known conflicts of interest associated with this publication and there has been no significant financial support for this work that could have influenced its outcome. RIFM staff are employees of the Research Institute for Fragrance Materials, Inc. (RIFM). The Expert Panel receives a small honorarium for time spent reviewing the subject work.

Objective: Prototype cosmetic formulations containing short-chain acids and alcohols intended to be applied in the proximity of the eyes are sometimes evaluated for ocular irritation potential using the validated Bovine Corneal Opacity and Permeability Assay (OECD TG 437). We evaluated the eye irritation potential of nine experimental cosmetic formulations designed and prepared by Avon Global Reserach and Development to differ only in the concentrations of Ethanol, Glycolic Acid and Salicylic Acid., Methods: We analysed the data generated using the BCOP assay. The opacity and permeability values obtained following the exposure of bovine corneas to experimental cosmetic formulations were combined into a single In Vitro Irritancy Score used to rank eye irritation potential. Histopathological examination of treated corneas was used to provide additional information about the depth and degree of the injury and to support the prediction of eye irritation potential of each experimental cosmetic formulation., Results: The In Vitro Irritancy Scores and histopathological analysis showed that experimental formulations containing only Ethanol, Glycolic Acid, or Salicylic Acid alone had, at most, a mild ocular irritation potential. The experimental formulations containing both Ethanol and Glycolic Acid had a mild ocular irritation potential, while the experimental formulations containing both Ethanol and Salicylic Acid had a moderate ocular irritation potential. Severe ocular irritation potential was induced by an experimental formulation containing a combination of Glycolic Acid and Salicylic Acid and it was further accentuated by the addition of Ethanol to the formulation. Our data indicate a possible synergistic effect on eye irritation potential of Ethanol, Glycolic Acid and Salicylic Acid in at least some experimental cosmetic formulations. Further, our results provide insight on an apparent concentration-dependent ocular irritation potential effect of combinations of Glycolic Acid, Salicylic Acid and Ethanol in at least one experimental cosmetic formulation., Conclusions: The results presented herein emphasise the need to consider in vitro testing of prototype cosmetic formulations containing combinations of Ethanol, Glycolic Acid and Salicylic Acid rather than relying on any predicted additive effect on ocular irritation based solely on previously generated results of similar formulations containing Ethanol, Glycolic Acid or Salicylic Acid alone. Further work is required to understand the significance of these observations and to elucidate the mechanisms responsible for the apparent synergistic effects of Glycolic Acid, Salicylic Acid and Ethanol and eye irritation potential suggested by our results.

The Expert Panel for Cosmetic Ingredient Safety (Panel) reassessed the safety of Capryloyl Salicylic Acid in cosmetic products; this ingredient is reported to function as a skin conditioning agent. The Panel reviewed relevant data relating to the safety of this ingredient in cosmetic formulations, and concluded that the available data are insufficient to make a determination that Capryloyl Salicylic Acid is safe under the intended conditions of use in cosmetic formulations., Competing Interests: Declaration of Conflicting InterestsThe author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Case studies are needed to demonstrate the use of human-relevant New Approach Methodologies in cosmetics ingredient safety assessments. For read-across assessments, it is crucial to compare the target chemical with the most appropriate analog; therefore, reliable analog selection should consider physicochemical properties, bioavailability, metabolism, as well as the bioactivity of potential analogs. To complement in vitro bioactivity assays, we evaluated the suitability of three potential analogs for the UV filters, homosalate and octisalate, according to their in vitro ADME properties. We describe how technical aspects of conducting assays for these highly lipophilic chemicals were addressed and interpreted. There were several properties that were common to all five chemicals: they all had similar stability in gastrointestinal fluids (in which no hydrolysis to salicylic occurred); were not substrates of the P-glycoprotein efflux transporter; were highly protein bound; and were hydrolyzed to salicylic acid (which was also a major metabolite). The main properties differentiating the chemicals were their permeability in Caco-2 cells, plasma stability, clearance in hepatic models, and the extent of hydrolysis to salicylic acid. Cyclohexyl salicylate, octisalate, and homosalate were identified suitable analogs for each other, whereas butyloctyl salicylate exhibited ADME properties that were markedly different, indicating it is unsuitable. Isoamyl salicylate can be a suitable analog with interpretation for octisalate. In conclusion, in vitro ADME properties of five chemicals were measured and used to pair target and potential analogs. This study demonstrates the importance of robust ADME data for the selection of analogs in a read-across safety assessment., (© 2024 John Wiley & Sons Ltd.)

Salicylic acid topical is used to treat variety of skin conditions. However, salicylic acid in these products is generated through industrial synthesis and has been shown to negatively impact fetal development and cause congenital abnormalities. We hypothesized that teratogenic effects reported in salicylic acid can be prevented by naturally synthesizing salicylic acid from wintergreen oil using green chemistry method. For this purpose, we investigated the effects of natural salicylic acid (NSA) synthesized from wintergreen oil using green chemistry and synthetic salicylic acid (SSA) on keratinocyte cell (HaCaT) proliferation and zebrafish embryo development. NSA structures were analyzed by 1 H NMR, 13 C NMR, and GC/MS methods. Percentage inhibition against HaCaT cell was determined by MTS assay. xCelligence system was used for cellular activities. Zebrafish embryos were exposed to NSA and SSA for 72 h post-fertilization. Lipid peroxidation, nitric oxide, sialic acid, glutathione-S-transferase, catalase, and superoxide dismutase were evaluated using biochemical methods. Expressions of nqO1 , gfap , bdnf , vtg , egr , cyp1a , and igf2 were determined by RT-PCR as developmental indicators. MTS and RT-cell analysis showed increased cell viability by NSA, whereas SSA decreased cell viability. NSA beneficially affected zebrafish embryo development while SSA exerted deleterious effects through oxidant-antioxidant status, inflammation, and development. Results of our study showed for the first time that synthesis of salicylic acid from wintergreen oil by green chemistry overcomes its cytotoxicity in keratinocyte cells and teratogenicity in zebrafish embryos. This finding is important for drug research on safe topical applications during pregnancy, when preventing exposure to drug and chemical-derived teratogens is vital.

The impact of pharmaceuticals on marine invertebrates has been a topic of rising concern, with an increasing number of studies regarding the impacts on bivalves. However, very few investigated the toxicity of mixtures of pharmaceuticals. This knowledge gap was investigated in the present study, where the toxicity of 17α-ethinylestradiol (EE2) and salicylic acid (SA) mixture was evaluated. To this end, Mytilus galloprovincialis mussels were chronically subjected to both pharmaceuticals, acting alone and in combination, and the effects at the cellular level were measured. The Independent Action (IA) model was performed aiming to compare obtained with predicted responses. The integrated biomarker response (IBR) index was used to assess the overall biochemical response given by mussels. The results obtained revealed that the most stressful condition was caused by the combined effect of EE2 and SA, with the highest metabolic capacity, antioxidant (catalase activity) and biotransformation (carboxylesterases activity) activation and cellular damage in organisms exposed to the mixture of both drugs in comparison to responses observed when each drug was acting alone. Predicted responses obtained from the IA model indicate that caution should be paid as frequent deviations to observed responses were found. This study highlights the need for future studies considering the mixture of pollutants, mimicking the actual environmental conditions., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Urban and hospital-sourced pharmaceuticals are continuously discharged into aquatic environments, threatening biota. To date, their impact as single compounds has been widely investigated, whereas few information exists on their effects as mixtures. We assessed the time-dependent biological impact induced by environmental concentrations of caffeine alone (CAF; 5 ng/L to 10 µg/L) and its combination with salicylic acid (CAF+SA; 5 ng/L+0.05 µg/L to 10 µg/L+100 µg/L) on gills of mussel Mytilus galloprovincialis during a 12-day exposure. Although no histological alteration was observed in mussel gills, haemocyte infiltration was noticed at T12 following CAF+SA exposure, as confirmed by flow cytometry with increased hyalinocytes. Both the treatments induced lipid peroxidation and cholinergic neurotoxicity, which the antioxidant system was unable to counteract. We have highlighted the biological risks posed by pharmaceuticals on biota under environmental scenarios, contributing to the enhancement of ecopharmacovigilance programmes and amelioration of the efficacy of wastewater treatment plants., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Abstract: Salicylic acid (SA) is one of the key signal molecules in regulating plant resistance to diverse pathogens. In Arabidopsis thaliana, it is predominantly associated with resistance against biotrophic and hemibiotrophic pathogens, and triggering systemic acquired resistance. In contrast, the effect of SA on the defence efficiency of wheat against fusarium head blight (FHB) and its causal agent, Fusarium graminearum, is still poorly understood. Here we show that the F. graminearum mycelial growth and conidia germination were significantly inhibited, and eventually halted in the presence of increasing concentration of SA in both liquid and solid media. Addition of SA also significantly reduced the production of the mycotoxin deoxynivalenol (DON). However the inhibitory effect of SA required acidic growth conditions to be observed while basic conditions allowed F. graminearum to use SA as a carbon source. High performance liquid chromatography (HPLC) analysis confirmed the capacity of F. graminearum to metabolize SA. To better understand the effect of SA on F. graminearum mycelial growth, we have compared the expression profiles of SA-treated and untreated F. graminearum liquid cultures after 8 and 24 h of treatment, using an F. graminearum custom-commercial microarray. The microarray analysis suggested that F. graminearum can metabolize SA through either the catechol or gentisate pathways that are present in some fungal species. Inoculation of F. graminearum conidia in a SA-containing solution has led to reduced FHB symptoms in the very susceptible Triticum aestivum cv. Roblin. In contrast, no inhibition was observed when SA and conidia were inoculated sequentially. The expression patterns for the wheat PR1, NPR1, Pdf1.2, and PR4 genes, a group of indicator genes for the defence response, suggested that SA-induced resistance contributed little to the reduction of symptoms in our assay conditions. Our results demonstrate that, although F. graminearum has the capacity to metabolize SA, SA has a significant and direct impact on F. graminearum through a reduction in efficiency of germination and growth at higher concentrations. [Copyright &y& Elsevier]

Despite the availability of analytic data, little is known about the toxicity of salicylic acid (SA) on aquatic non-target organisms. The present study aimed at evaluating the impact of SA through a short-term exposure of the Mediterranean mussel Mytilus galloprovincialis to five environmentally relevant concentrations of SA. A set of suitable biomarkers was applied at selected time-points on mussel digestive glands, including histological observations and expression of oxidative stress related genes. The obtained results showed a conspicuous hemocytic infiltration among mussel digestive tubules, as confirmed also by a flow cytometric approach that revealed an increase of halinocytes and granulocytes. Interestingly, a significant dose and time dependent decrease in the expression levels of oxidative stress related genes was found in mussels exposed to SA except for the glutathione S-transferase gene that was significantly up-regulated in a time-dependent manner confirming its important role against oxidant species and in the metabolism of pharmaceuticals., (Copyright © 2022 Elsevier B.V. All rights reserved.)

The main objective of the study was to assess if joint application of melatonin (MT, 0.1 mM) and salicylic acid (SA 0.5 mM) could improve tolerance of pepper plants to arsenic (As) as sodium hydrogen arsenate heptahydrate (0.05 mM). The imposition of arsenic stress led to accumulation of As in roots and leaves, and increased contents of leaf proline, phytochelatins, malondialdehyde (MDA) and H 2 O 2 , but it reduced plant biomass, chlorophylls (Chl), PSII maximum efficiency (Fv/Fm) and leaf water potential. Melatonin and SA applied jointly or alone enhanced nitrogen metabolism by triggering the activities of glutamate synthase, glutamine synthetase, and nitrite reductases and nitrate. In comparison with a single treatment of MT or SA, the joint treatment of MT and SA had better impact on enhancing growth and key biological events and decreasing tissue As content. This clearly shows a cooperative function of both agents in enhancing tolerance to As-toxicity in pepper plants., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

The increasing presence of anthropogenic contaminants in the environment may constitute a challenge to non-target biota, considering that most contaminants can exert deleterious effects. Salicylic acid (SA) is a non-steroid anti-inflammatory drug (NSAID) which exerts its activity by inhibiting the enzyme cyclooxygenase (COX). Another class of drugs is that of the diuretics, in which acetazolamide (ACZ) is included. This pharmaceutical acts by inhibiting carbonic anhydrase (CA), a key enzyme in acid-base homeostasis, regulation of pH, being also responsible for the bio-availability of Ca 2+ for shell biomineralization processes. In this work, we evaluated the chronic (28-day) ecotoxicological effects resulting from the exposures to SA and ACZ (alone, and in combination) on individuals of the marine mussel species Mytillus spp., using enzymatic (catalase (CAT), glutathione S-transferases (GSTs), COX and CA), non-enzymatic (lipid peroxidation, TBARS levels) and morphological and physiological (shell hardness, shell index and feeding behaviour) biomarkers. Exposure to ACZ and SA did not cause significant alterations in CAT and GSTs activities, and in TBARS levels. In terms of CA, this enzyme was inhibited by the highest concentration of ACZ in gills of exposed animals, but no effects occurred in the mantle tissue. The activity of COX was not altered after exposure to the single chemicals. However, animals exposed to the mixture of ACZ and SA evidenced a significant inhibition of COX activity. Morphological and physiological processes (namely, feeding, shell index, and shell hardness) were not affected by the here tested pharmaceutical drugs. Considering the general absence of adverse effects, further studies are needed to fully evaluate the effects of these pharmaceutical drugs on alternative biochemical and physiological pathways., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Boron (B) toxicity is a notable abiotic hindrance that restricts crop productivity by disturbing several physiological and biochemical processes in plants. This study was aimed to elucidate the role of salicylic acid (SA) in conferring tolerance to B stress in Mentha arvensis and Cymbopogon flexuosus. Boron toxicity led to a considerable decrease in shoot height and root length, fresh and dry mass of shoot and root, and physiological and biochemical parameters. However, exogenously applied SA relieved the adverse effects caused by B toxicity and led to an increase in growth parameters under B stress and non-stress conditions. The treatment of B resulted in its increased accumulation in roots and shoots of both the plants which, in turn, caused oxidative damage as evident by increased content of malondialdehyde and catalase, peroxidase, superoxide dismutase and glutathione reductase enzyme activities. However, exogenous SA supply significantly affected antioxidant enzyme activities and protected the plants from excess B. Moreover, the essential oil content of two selected plants declined under B toxicity and significantly enhanced in SA-treated stressed plants. The contents of menthol and menthyl acetate in M. arvensis were lowered in B stressed plants which significantly improved in SA treated B-stressed and in their respective SA alone treatment. Similarly, citral-A and citral-B content of C. flexuosus declined under B toxicity, however, SA reversed the negative effects of B toxicity on essential oil components. This assessment stipulated the promising role of exogenously applied SA in alleviating B toxicity in M. arvensis and C. flexuosus by improving antioxidant machinery and limiting B uptake which protects the structural integrity of leaves and also helps in increasing essential oil content., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Salicylic acid (SA), a metabolite of acetylsalicylic acid is a monohydroxybenzoic acid a common non-steroidal analgesic and anti-inflammatory drug (NSAID) frequently detected in various aquatic ecosystems at concentrations up to 19.50 μg L -1 in surface waters near livestock farms and 59.6 μg L -1 in wastewaters. Little is known on the effects of short-term exposure of freshwater crustaceans to salicylic acid. Therefore, the aim of our study was to determine the effects of SA at concentrations of 5 μg L -1 , 500 μg L -1 , 5 mg L -1 , 50 mg L -1 and 500 mg L -1 on the behavior (swimming speed, swimming height, distance travelled) and physiological endpoints (heart rate, mandible movement) of Daphnia magna exposed for 24 h, 48 h and 72 h. The results showed that SA inhibited the swimming speed, swimming height and distance travelled, heart rate and mandible movement at 5 mg L -1 , 50 mg L -1 and 500 mg L -1 when compared to the control. On the other hand, SA at 5 μg L -1 and 500 μg L -1 transiently increased swimming speed and distance travelled after 24 h of the exposure, except for swimming height. Behavioral and physiological disturbances were observed much earlier than lethality. Our study showed SA at environmental levels may be an ecotoxicological agent imparing behavior and physiology of freshwater crustaceans., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

The role of nitric oxide (NO) in salicylic acid (SA)-induced tolerance to arsenic (As) stress in maize plants is not reported in the literature. Before starting As stress (AsS) treatments, SA (0.5 mM) was sprayed to the foliage of maize plants. Thereafter, AsV (0.1 mM as sodium hydrogen arsenate heptahydrate) stress (AsS) was initiated and during the stress period, sodium nitroprusside (SNP 0.1 mM), a NO donor, was sprayed individually or in combination with SA. Furthermore, cPTIO (0.1 mM) was also applied as a NO scavenger during the stress period. Arsenic stress led to significant reductions in plant growth, photosynthesis, water relation parameters and endogenous NO content, but it increased hydrogen peroxide, malondialdehyde, electrolyte leakage, methylglyoxal, proline, the activities of major antioxidant enzymes, and leaf and root As content. The combined treatment of SA+SNP was more effective to reverse oxidative stress related parameters and reduce the As content in both leaves and roots, with a concomitant increase in antioxidant defense system, the ascorbate-glutathione (AsA-GSH) cycle-related enzymes, glyoxalase system enzymes, plant growth, and photosynthetic traits. The beneficial effects of SA were completely abolished with cPTIO supply by blocking the NO synthesis in AsS-maize plants, indicating that NO effectively participated in SA-improved tolerance to AsS in maize plants., (Copyright © 2020 Elsevier B.V. All rights reserved.)

While many studies have been conducted on drug-inducing alterations in the aquatic environment, little is known about their interaction with climate change, such as rising temperatures. To increase knowledge on this topic, Mytilus galloprovincialis mussels were exposed to two different temperatures 17 ± 1 °C (control) and 21 ± 1 °C in the absence and presence of salicylic acid (SA) (4 mg/L) for 28 days. Salicylic acid in the water and tissues was measured and its impact reported through biomarker responses including: energy metabolism (electron transport system (ETS) activity, glycogen (GLY), protein (PROT) and lipids (LIP) contents), oxidative stress markers (activity of the enzymes superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx)), glutathione balance between the reduced and the oxidized forms (GSH/GSSG), and damage to membrane lipids (lipid peroxidation - LPO). The mussels responded differently if the stresses imposed were single or combined, with greater impacts when both stressors were acting together. Contaminated mussels exposed to high temperatures were unable to increase their metabolic capacity to restore their defence mechanisms, reducing the expenditure of LIP. In the presence of SA and increased temperature antioxidant defences respond differently, with higher SOD levels and inhibition of CAT. The present study highlights not only the negative impact of warming and SA, but especially how temperature increase will promote the impact of SA in M. galloprovincialis, which under predicted climate change scenarios may greatly impair population maintenance and ecosystem biodiversity., (Copyright © 2020 Elsevier B.V. All rights reserved.)

We investigated the mechanistic consequences of selenium (Se)-toxicity, and its possible mitigation using salicylic acid (SA) in rice. In comparison with control, sodium selenate-exposed 'Se1' (0.5 mM) and 'Se2' (1.0 mM) plants showed accumulation of Se by 190.63 and 288.00 % in roots, 2359.42 and 2054.35 % in leaf sheaths, and 7869.91 and 9063.72 % in leaves, respectively, resulting in severe toxicity symptoms, such as growth inhibition, chlorosis, burning of leaves, and oxidative stress. In contrast, SA addition to Se-stressed plants significantly alleviated the Se-toxicity symptoms, and radically improved shoot height (28.88 %), dry biomass (34.00 %), total chlorophyll (37.51 %), soluble sugar (17.31 %) and leaf water contents (22.31 %) in 'SA + Se2' plants over 'Se2' plants. Notably, SA maintained Se-homeostasis, and decreased 'Se2'-induced oxidative stress by enhancing ascorbate level (67.75 %) and the activities of antioxidant enzymes like superoxide dismutase (20.99 %), catalase (40.97 %), glutathione peroxidase (12.26 %), and glutathione reductase (32.58 %) relative to that in 'Se2' plants. Additionally, SA protected rice plants from the deleterious effects of methylglyoxal by stimulating the activities of glyoxalase enzymes. Furthermore, SA upregulated several genes associated with reactive oxygen species (e.g. OsCuZnSOD1, OsCATB, OsGPX1 and OsAPX2) and methylglyoxal (e.g. OsGLYI-1) detoxifications. These findings unravel a decisive role of SA in alleviating Se-phytotoxicity in rice., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Alterations of the physical-chemical properties of the oceans due to anthropogenic activities are, at present, one of the most concerning environmental issues studied by researchers. One of these issues is ocean acidification, mainly caused by overproduction and release of carbon dioxide (CO 2 ) from anthropogenic sources. Another component of environmental degradation is related to the production and release of potential toxic compounds, namely active pharmaceutical ingredients, into the aquatic environment that, combined with oceanic acidification, can cause unpredictable and never before considered deleterious effects on non-target marine organisms. Regarding this issue, the hereby study used predictions of future ocean acidification to simulate realistic scenarios of environmental exposure to a common therapeutic drug, salicylic acid (SA), in the marine gastropod Gibbula umbilicalis under different pH values. This species was exposed to a range of pH values (8.2, 7.9 and 7.6), and to already reported environmentally realistic concentrations (5, 25 and 125 μg/L) of SA. To evaluate the effects of these environmental stressors, key physiological biomarkers (GSTs, CAT, TBARS, AChE and COX) and shell hardness (SH) were quantified. Results from the present study showed that CAT and GSTs activities were enhanced by SA under water acidification; increased lipid peroxidation was also observed in organisms exposed to SA in more acidic media. In addition, the hereby study demonstrated the neurotoxic effects of SA through the inhibition of AChE. Effects were also observed in terms of COX activity, showing that SA absorption may be affected by water acidification. In terms of SH, the obtained data suggest that SA may alter the physical integrity of shells of exposed organisms. It is possible to conclude that the combination of seawater acidification and exposure to toxic xenobiotics (namely to the drug SA) may be strenuous to marine communities, making aquatic biota more susceptible to xenobiotics, and consequently endangering marine life in an unpredictable extent., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

In the present study, the biochemical responses and antioxidant enzymes activity of the Dunaliella salina, a green microalga, to the interaction of silver nanoparticles (AgNPs) and salicylic acid (SA) were investigated. Algal suspensions in the phase of logarithmic growth were subjected to the concentrations of 0, 5, 15, and 25 pM AgNPs with or without 1 mM SA. AgNPs level of 25 pM declined cell division but highly accumulated levels of chlorophyll, β-carotene, proteins, free amino acid, carbohydrates, and hydrogen peroxide, which was associated with enhanced the activity of proteolysis, lipid peroxidation, and antioxidant enzymes. SA-treated cells at 25 pM AgNPs improved cell growth but declined the activities of antioxidant enzymes and proteolytic along with a lower accumulation of metabolites except β-carotene relative to untreated controls. These results suggest that AgNPs treatment induce oxidative stress in D. salina cells, which tolerated by alga through the metabolic modifications and accumulating β-carotene, while SA induces AgNPs tolerance by the mechanisms that direct carbon flux to growth and β-carotene biosynthesis rather than the antioxidant enzymes or osmoprotectant metabolites., (Copyright © 2019 Elsevier B.V. All rights reserved.)

The global occurrence of pharmaceuticals in the aquatic environment has been considered a particularly concerning problem with unknown consequences. Non-steroidal anti-inflammatory drugs (NSAIDs) including diclofenac (DCF) and salicylic acid (SA), are among the most frequently prescribed drugs in the world, being consequently commonly found in the aquatic environment. Prolonged experiments (with duration of exposure that surpass those recommended by already established testing guidelines) are important to obtain ecologically relevant data to address the issue of NSAIDs ecotoxicity, because by being more realistically (namely in terms of levels and durations of exposure), such tests may indicate realistic challenges posed to aquatic organisms. Among the most common test species that are used for assessing environmental quality, plants play a leading role. Lemna species are among the most important plants used for ecotoxicity testing. Therefore, the aim of this study was to evaluate the temporal effect of a prolonged exposure of DCF and SA on Lemna minor. To attain this purpose, L. minor plants were chronically exposed to 0, 4, 20, and 100 μg/L of both pharmaceuticals, and samplings were performed at 6, 10 and 14 days of exposure. The analyzed endpoints were: levels of chlorophyll a, b and total, carotenoids; and enzymatic biomarkers, such as catalase, ascorbate peroxidase and glutathione-S-transferases. Diclofenac was responsible for alterations in all analyzed parameters in different intervals of exposure. Salicylic acid exposure was not capable of causing alterations on pigment contents of L. minor, however, enzymatic biomarkers were altered at all sampling intervals. Thus, it is possible to conclude that both pharmaceuticals can cause damage on the tested macrophyte species, biochemical parameters being more sensitive than physiological ones. Additional prolonged experiments are required to understand the chronic effects of different pharmaceuticals in the aquatic environment, especially in plants., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Background: Asthma exacerbations evoke emergency room visits, progressive loss of lung function and increased mortality. Environmental and industrial toxicants exacerbate asthma, although the underlying mechanisms are unknown. We assessed whether 3 distinct toxicants, salicylic acid (SA), toluene diisocyanate (TDI), and 1-chloro-2,4-dinitrobenzene (DNCB) induced airway hyperresponsiveness (AHR) through modulating excitation-contraction coupling in human airway smooth muscle (HASM) cells. The toxicants include a non-sensitizing irritant (SA), respiratory sensitizer (TDI) and dermal sensitizer (DNCB), respectively. We hypothesized that these toxicants induce AHR by modulating excitation-contraction (EC) coupling in airway smooth muscle (ASM) cells., Methods: Carbachol-induced bronchoconstriction was measured in precision-cut human lung slices (hPCLS) following exposure to SA, TDI, DNCB or vehicle. Culture supernatants of hPCLS were screened for mediator release. In HASM cells treated with the toxicants, surrogate readouts of EC coupling were measured by phosphorylated myosin light chain (pMLC) and agonist-induced Ca 2+ mobilization ([Ca 2+ ] i ). In addition, Nrf-2-dependent antioxidant response was determined by NAD(P) H quinone oxidoreductase 1 (NQO1) expression in HASM cells., Results: In hPCLS, SA, but not TDI or DNCB, potentiated carbachol-induced bronchoconstriction. The toxicants had little effect on release of inflammatory mediators, including IL-6, IL-8 and eotaxin from hPCLS. In HASM cells, TDI amplified carbachol-induced MLC phosphorylation. The toxicants also had little effect on agonist-induced [Ca 2+ ] i. CONCLUSION: SA, a non-sensitizing irritant, amplifies agonist-induced bronchoconstriction in hPCLS via mechanisms independent of inflammation and Ca 2+ homeostasis in HASM cells. The sensitizers TDI and DNCB, had little effect on bronchoconstriction or inflammatory mediator release in hPCLS., Implications: Our findings suggest that non-sensitizing irritant salicylic acid may evoke AHR and exacerbate symptoms in susceptible individuals or in those with underlying lung disease.

Contamination of the aquatic environment by pharmaceutical drugs is an emerging issue in ecotoxicology. Aquatic organisms, in the presence of xenobiotics, tend to activate defensive mechanisms against toxic effects in order to mitigate and/or compensate for the toxic damages that frequently result from these interactions. Salicylic acid (SA) is a common drug, widely used in human medicine due to its analgesic, anti-inflammatory, and antipyretic properties, as well as its activity in terms of preventing platelet aggregation, among other clinical and cosmetic uses. It is commonly found in levels of the nanograms per liter to the micrograms per liter range in receiving waters, and its presence has been related to toxic effects in aquatic organisms, including oxidative stress. However, the number of studies that characterize the ecotoxicological profile of salicylates is still scarce and no studies have been published about the putative toxic effects of SA, especially in marine polychaetes. In order to determine the potential ecotoxicological effects caused by SA, individuals of the marine Polychaeta species Hediste diversicolor were exposed for 96 h to ecologically relevant concentrations of this compound, and several biochemical endpoints were evaluated, namely the activity of the antioxidant enzymes glutathione peroxidase (GPx) and catalase (CAT), the phase II biotransformation isoenzymes glutathione S-transferases (GSTs), the cholinergic enzyme acetylcholinesterase (AChE), and the determination of lipoperoxidative damage (thiobarbituric acid-reactive substances (TBARS) assay). The obtained results demonstrated that despite the pro-oxidative effects elicited by SA, exposure to realistic levels of this compound was not able to generate a state of oxidative stress, and the adaptive protective responses elicited by exposed individuals were effective enough to minimize and/or inhibit the damage potentially caused by overproduced reactive oxygen species.

The technological utility of active pharmaceutical ingredients (APIs) is greatly enhanced when they are transformed into ionic liquids (ILs). API-ILs have better solubility, thermal stability, and the efficacy in topical delivery than solid or crystalline drugs. However, toxicological issue of API-ILs is the main challenge for their application in drug delivery. To address this issue, 11 amino acid esters (AAEs) were synthesized and investigated as biocompatible counter cations for the poorly water-soluble drug salicylic acid (Sal) to form Sal-ILs. The AAEs were characterized using 1 H and 13 C NMR, FTIR, elemental, and thermogravimetric analyses. The cytotoxicities of the AAE cations, Sal-ILs, and free Sal were investigated using mammalian cell lines (L929 and HeLa). The toxicities of the AAE cations greatly increased with inclusion of long alkyl chains, sulfur, and aromatic rings in the side groups of the cations. Ethyl esters of alanine, aspartic acid, and proline were selected as a low cytotoxic AAE. The cytotoxicities of the Sal-ILs drastically increased compared with the AAEs on incorporation of Sal into the cations, and were comparable to that of free Sal. Interestingly, the water miscibilities of the Sal-ILs were higher than that of free Sal, and the Sal-ILs were miscible with water at any ratio. A skin permeation study showed that the Sal-ILs penetrated through skin faster than the Sal sodium salt. These results suggest that AAEs could be used in biomedical applications to eliminate the use of traditional toxic solvents for transdermal delivery of poorly water-soluble drugs., (Copyright © 2018 Elsevier B.V. All rights reserved.)

New series of oxo-vanadium N-salicyledieneamino acid Schiff base complexes are synthesized and characterized. They are synthesized from the reaction of sodium salicylaldehyde-5-sulfonate, some amino acids, alanine (VOHL1), leucine (VOHL2) or glycine (VOHL3) in an aqueous media, and leucine (VOHLpy1) or tryptophan (VOHLpy2) in pyridine with vanadyl acetylacetonate. The complexes are characterized by EA, TGA, IR, UV-Visible and mass spectra, conductivity and magnetic measurements. The biological activity of the VO-complexes shows that VOHL1, VOHL2 and VOHL3 exhibit anti-proliferative effect and may be used as anticancer drugs. VO-complexes manifest high toxicity, except VOHL2 is less toxic, and could be applied for the human being. VOHL1, VOHL2 and VOHL3 display remarkable SOD like potential and act as high inhibiting reagents. VOHLpy1 and VOHLpy2 show low inhibiting potentials. VO-complexes have good anti-oxidant effect, in which VOHL3 affords the best antioxidant activity. The interaction between VO-complexes and DNA is studied spectrophotometrically and by gel electrophoresis. Binding constants and spectrophotometric parameters indicate a strong interaction between VO-complexes and DNA. VO-complexes have respectable anti-bacterial and antifungal activities, where VOHL3 shows the maximum potential. DFT calculations of VOHL1 and VOHL3 were discussed in the light of their biological activity, which are convenient with the obtained results., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Salicylic acid (SA) has a long history of safe use as ingredient in topical cosmetic products. In 2016, the Committee for Risk Assessment of the European Chemicals Agency proposed to classify SA as a Category 2 reproductive toxicant based on adverse developmental effects in animal toxicity studies. This hazard-based classification (based on mg/kg doses) requires a reassessment of the safety of the current SA concentrations in cosmetic consumer products. Herein, a safety reassessment was performed in which margins of safety were calculated based on literature data on the NOAEL plasma exposure levels from animal reproductive toxicity studies with ASA (rapidly converts to SA in plasma), human SA plasma levels from oral exposure to ASA and human dermal exposure to SA-containing cosmetic products. In addition, a literature review was performed, which shows that there are no adverse developmental effects despite extensive human clinical oral use of ASA up to the maximum recommended therapeutic doses. The plasma exposure-based safety assessment for SA combined with an absence of any clinical health risk with oral ASA use in the literature supports that there is an acceptable margin of safety for the consumer exposure to SA as authorized in the current EU cosmetic regulation., (Copyright © 2018 The Authors. Published by Elsevier Inc. All rights reserved.)

The neutral red uptake assay is a cell viability assay that allows in vitro quantification of xenobiotic-induced cytotoxicity. The assay relies on the ability of living cells to incorporate and bind neutral red, a weak cationic dye, in lysosomes. As such, cytotoxicity is expressed as a concentration-dependent reduction of the uptake of neutral red after exposure to the xenobiotic under investigation. The neutral red uptake assay is mainly used for hazard assessment in in vitro toxicology applications. This method has also been introduced in regulatory recommendations as part of 3T3-NRU-phototoxicity-assay, which was regulatory accepted in all EU member states in 2000 and in the OECD member states in 2004 as a test guideline (TG 432). The present protocol describes the neutral red uptake assay using the human hepatoma cell line HepG2, which is often employed as an alternative in vitro model for human hepatocytes. As an example, the cytotoxicity of acetaminophen and acetyl salicylic acid is assessed.

Nonsteroidal anti-inflammatory drugs (NSAIDs) are widely used therapeutic agents; however, their pharmacological actions raise concerns about potential risks to the reproductive health of aquatic vertebrates. In the present study, a medaka ovulation assay was applied as an in vitro model to evaluate NSAID-induced antiovulatory activity. We first tested five NSAIDs, including diclofenac sodium (DCF), ketoprofen (KP), salicylic acid (SA), mefenamic acid (MA), and acetylsalicylic acid (ASA) for their antiovulatory activities toward the follicles isolated from the ovaries of spawning females. Of all the chemicals tested, DCF had the highest antiovulatory activity, with the concentration that caused 50% inhibition (IC50) (101 µM). MA was the second most potent inhibitor following DCF, but KP, SA, or ASA had little inhibitory effect on the ovulation of the follicles. The in vitro antiovulatory activity of five NSAIDs showed good correlation with data published on the inhibitory activity on human COX-2. Second, we selected DCF and SA as the most and least potent NSAIDs, respectively, and examined the effects on reproduction of intact fish in order to evaluate whether the ovulation assay was a reasonable predictor of potential reproductive effects in fish. Females exposed to DCF showed a concentration-dependent decrease in the number of spawned eggs and an increment in the gonadosomatic index (GSI), possibly due to an anovulation in the females. In contrast, neither fecundity nor the GSI of females decreased at up to 20 mg/L of SA, at which acute lethality to medaka was induced. In conclusion, the medaka ovulation assay reflected the potency of NSAID-induced antiovulatory activity and may thus serve as an in vitro model for the prediction of NSAID-induced reproductive toxicity. © 2015 Wiley Periodicals, Inc. Environ Toxicol 31: 1710-1719, 2016., (© 2015 Wiley Periodicals, Inc.)

The aquatic environment is becoming increasingly contaminated with pharmaceuticals. Salicylic acid (SA), which can be used individually or appear as a degradation product of the widely used acetylsalicylic acid was chosen for testing. Juvenile zebrafish Danio rerio were subjected to OECD test No. 215 (fish, juvenile growth test) with salicylic acid concentrations of 0.004; 0.04; 0.4; 4 and 40 mg/L. Specific growth rate (SGR), histological changes, and parameters of oxidative stress were evaluated. SA had no effects on histological changes, SGR, glutathione reductase, and lipid peroxidation. Increased catalytic activity of GPx was found at 0.04 mg/L compared to control, increased catalytic activity of catalase was found at 0.04 and 4 mg/L compared to control, and increased catalytic activity of glutathione-S-transferase was found at 0.004 and 0.04 mg/L compared to control (P < 0.05). Juvenile zebrafish turned out to be relatively insensitive to both environmentally relevant (0.004 mg/L) and higher concentrations of salicylic acid.

Fusarium graminearum is the major causal agent of Fusarium head blight (FHB) of wheat and barley and is considered to be one of the most devastating plant diseases worldwide. Chitin is a critical component of the fungal cell wall and is polymerized from UDP-N-acetyl-alpha-D-glucosamine by chitin synthase. We characterized FgCHS8, a new class of the chitin synthase gene in F. graminearum. Disruption of FgCHS8 resulted in reduced accumulation of chitin, decreased chitin synthase activity, and had no effect on conidia growth when compared with the wild-type isolate. ΔFgCHS8 had a growth rate comparable to that of the wild-type isolate in vitro. However, ΔFgCHS8 had reduced growth when grown on agar supplemented with either 0.025% SDS or 0.9 mM salicylic acid. ΔFgCHS8 produced significantly less deoxynivalenol and exhibited reduced pathogenicity in wheat spikes. Re-introduction of a functional FgCHS8 gene into the ΔFgCHS8 mutant strain restored the wild-type phenotypes. Fluorescence microscopy revealed that FgCHS8 protein was initially expressed in the septa zone, and then gradually distributed over the entire cellular membrane, indicating that FgCHS8 was required for cell wall development. Our results demonstrated that FgCHS8 is important for cell wall sensitivity to environmental stress factors and deoxynivalenol production in F. graminearum., (Copyright © 2016 The British Mycological Society. Published by Elsevier Ltd. All rights reserved.)

A novel total toxicity detection method based on a mediated biosensor system with flow injection (MB-FI) was developed to rapidly and reliably detect respiration inhibitors (i.e., As2O3, KCN, salicylic acid (SA), 2,4-dintirophenol (DNP)) in water. The mediated biosensor toxicity assessment using microorganisms immobilized in calcium alginate filaments can greatly simplify the testing process and save time. In the MB-FI system, ferricyanide together with a respiration inhibitor was injected into the bioreactor, inhibiting the respiration of the immobilized microorganisms. The degree of inhibition was measured by determining the ferrocyanide generated in the effluent, expressed as the 50% inhibition concentration (IC50). The IC50 values for the four respiration inhibitors obtained using this method were comparable to those obtained using the classic method, confirming that this approach is an alternative alert method. More importantly, this constructed biosensor system with flow injection will facilitate the application and commercialization of this toxicity monitoring technology., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Environmental concentrations of pharmaceutical residues are often low; nevertheless, they are designed to have biological effects at low doses. The aim of this study was to assess the effects of salicylic acid on the growth and development of common carp (Cyprinus carpio) early life stages with respect to antioxidant defence enzymes. An embryo-larval toxicity test lasting 34 days was performed according to OECD guidelines 210 (Fish, Early-life Stage Toxicity Test). The tested concentrations were 0.004, 0.04, 0.4, 4 and 20mg/l of salicylic acid. Hatching, early ontogeny, and both morphometric and condition characteristics were significantly influenced by subchronic exposure to salicylic acid. Also, changes in antioxidant enzyme activity and an increase in lipid peroxidation were observed. The LOEC value was found to be 0.004 mg/l salicylic acid. The results of our study confirm the suggestion that subchronic exposure to salicylic acid at environmental concentrations can have significant effects on aquatic vertebrates., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Biotic stress factors such as Rhizopus oligosporus and Aspergillus niger mycelial extracts and abiotic elements methyljasmonate (MJ) and salicylic acid (SA), when administered through floral spray to Coffea canephora, showed significant influence on major bioactive metabolites of beans. Up to 42% caffeine, 39% theobromine and 46% trigonelline, along with 32% cafestol and kahweol content elevation was evident under respective elicitor treatments. Over all, the surge in respective metabolites depends on elicitor stress type and concentration. Abiotic factors MJ and SA were found to be efficient at 1 to 5 microM concentration in augmenting all the metabolites, compared to R. oligosporus and A. niger spray at 0.5-2.0% wherein the response was moderate as compared to abiotic stress, however significant compared to control. Though this elevation in caffeine, theobromine, cafestol and kahweol is not warranted from quality point of view, increase in trigonelline improves coffee quality. Besides increase in metabolites, stress mediated augmentation of bioactive compounds in coffee has a wide scope for studying gene expression pattern.

The presence of pharmaceutical drugs in aquatic ecosystems has been widely reported during the past years. Salicylic acid (SA) is mainly used in human medicine as an analgesic and antipyretic drug, being also active in preventing platelet aggregation. To study the ecotoxicological effects potentially elicited by SA in freshwater fish, brown trout individuals (Salmo trutta fario) were chronically exposed (28 days) to this drug, in order to evaluate the enzymatic and histological effects, in both gills and liver. A qualitative and semi-qualitative evaluation of the gills and liver was performed, and also a quantitative evaluation of various lamellar structures. Oxidative stress was quantified trough the determination of glutathione S-transferases (GSTs), glutathione reductase (GRed), total and selenium-dependent glutathione peroxidase (GPx) and Catalase (Cat) activities. Lipid peroxidative damage was also assessed by the quantification of thiobarbituric acid reactive substances (TBARS) in the liver. The here-obtained data showed the occurrence of oxidative stress, reflected by an increased activity of GPx and GRed in the liver; additionally, it was possible to observe non-specific histological changes in gills. The global significance of the entire set of results is discussed, giving emphasis to the ecological relevance of the responses.

Starvation and changing feeding conditions are frequently characteristics of wastewater treatment plants. They are typical causes of unsteady-state operation of biological systems and provoke cellular stress. The response of a membrane bioreactor functioning under feed-induced stress conditions is studied here. In order to simplify and considerably amplify the response to stress and to obtain a reference model, a pure culture of Pseudomonas putida was selected instead of an activated sludge and a sole substrate (salicylic acid) was employed. The system degraded salicylic acid at 100-1100mg/L with a high level of efficiency, showed rapid acclimation without substrate or product inhibition phenomena and good stability in response to unsteady states caused by feed variations. Under starvation conditions, specific degradation rates of around 15mg/gh were achieved during the adaptation of the biomass to the new conditions and no biofilm formation was observed during the first days of experimentation using an initial substrate to microorganisms ratio lower than 0.1. When substrate was added to the reactor as pulses resulting in rapidly changing concentrations, P. putida growth was observed only for substrate to microorganism ratios higher than 0.6, with a maximum YX/S of 0.5g/g. Biofilm development under changing feeding conditions was fast, biomass detachment only being significant for biomass concentrations on the membrane surface that were higher than 16g/m(2)., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Aspirin and its active ingredient salicylate are potent antioxidants that have been reported to be neuro- and otoprotective. However, when consumed in large quantities, these drugs can cause temporary hearing loss and tinnitus. Moreover, recent studies indicate that after several days of treatment, salicylate selectively destroys the spiral ganglion neurons and auditory nerve fibers that relay sounds from the sensory hair cells to the brain. Why salicylate selectively damages spiral ganglion neurons while sparing the hair cells and supports cells is unclear. Here we show that high dose of salicylate trigger an apoptotic response in spiral ganglion neurons characterized morphologically by soma shrinkage and nuclear condensation and fragmentation plus activation of extrinsic initiator caspase-8 and intrinsic initiator caspase-9 several days after the onset of drug treatment. Salicylate treatment triggered an upsurge in the toxic superoxide radical only in spiral ganglion neurons, but not in neighboring hair cells and support cells. Mn TMPyP pentachloride, a cell permeable scavenger of superoxide blocked the expression of superoxide staining in spiral ganglion neurons and almost completely blocked the damage to the nerve fibers and spiral ganglion neurons. NMDA receptor activation is known to increase neuronal superoxide levels. Since NMDA receptors are mainly found on spiral ganglion neurons and since salicylate enhances NMDA receptor currents, the selective killing of spiral ganglion neurons is likely a consequence of enhanced and sustained activation of NMDA receptors by salicylate.

Aims: Aging and drug-induced side effects may contribute to deteriorate mitochondrial bioenergetics in many tissues, including kidney and liver. One possibility is that the combination of both aging and drug toxicity accelerates the process of mitochondrial degradation, leading to progressive bioenergetic disruption. We therefore analyzed in vitro kidney (KM) and liver (LM) mitochondrial response to salicylate and diclofenac in old and adult animals., Main Methods: Male-Wistar adult (19-wks) and aged (106-wks) rats were used. In vitro endpoints of oxygen consumption and membrane potential were evaluated in non-treated conditions (vehicle) and in the presence of salicylate (0.5mM) and diclofenac (50μM). The susceptibility to calcium-induced permeability transition pore (MPTP) was assessed. Aconitase and C, -SH and MDA contents were measured. Apoptotic signaling was followed by measuring caspase 3, 8 and 9 activities, Bax, Bcl2 and CypD expression. ANT content was semi-quantified., Key Findings: In general, animal age alone compromised KM state 3 and LM ADP lag phase while resulting in decreased resistance to the MPTP. Aging decreased LM CypD and increased Mn-SOD. Kidney caspase 9-like activity was lower in aged group. Salicylate and diclofenac induced KM and LM dysfunction. ADP lag phase in KM was further increased in the aged group in the presence of diclofenac. No further impairments were observed regarding drug toxicity adding to the aging process., Significance: Aging impaired KM and LM function despite no detected alterations on oxidative stress and apoptosis. However, aging did not further exacerbate KM and LM frailty induced by salicylate and diclofenac., (© 2013.)

Rats were trained in a two lever food reinforced operant procedure to discriminate a 8000 Hz pure tone stimulus from its absence. Responding on one lever was reinforced in the presence of the tone and responding on the other lever was reinforced when the tone was absent. Frequency generalization testing yielded an inverted U-shaped function, whereas sound pressure level generalization testing yielded a continuous decrease in responding on the tone associated lever with decreasing sound pressure levels. The administration of sodium salicylic acid (150-450 mg/kg) generated responding on the tone associated lever suggesting that salicylic acid induced an experience that had commonalities with the percept of the training tone stimulus. After exposure to intense sound, responding consistent with the presence of tinnitus was achieved and Lidocaine failed to reduce tinnitus behavior. The use of a two choice design helped avoid confounding factors induced by drug induced side effects. Further, since no auditory cues were employed in the test situation the model achieves resistance to potential bias due to hearing impairment and hyperacusis. We propose that this model may be useful in detecting tinnitus., (Copyright © 2013 Elsevier B.V. All rights reserved.)

This study focused on fabrication and characterization of porous core/sheath structured composite nanofibers with a core of blended salicylic acid (SA) and poly(ethylene glycol) (PEG) and a sheath of poly(lactic acid) (PLA) using a dual-capillary electrospinning system. Results of water contact angle measurements, field-emission scanning electron microscopy, and transmission electron microscopy indicated that feed rates of the core and sheath strongly affect the stability of the core/sheath structure and porous density of the composite nanofibers obtained, significantly influencing their SA release characteristics. At a lower ratio of feed rates of the core and the sheath, better stable core/sheath structures of nanofibers with higher porous density on the surface were formed resulting in a sustained release of SA over 5 days. Non-porous fibers showed a lower amount of drug release because the drug was embedded inside the core layer of the non-porous sheath layer. SA release from porous core/sheath nanofibers was described based on a one-dimensional Fickian diffusion mechanism, indicating that drug diffusion is a predominant factor in drug release. A cytotoxicity test suggested that the porous core/sheath nanofibers are non-toxic and support cell attachment. Therefore, this fiber mat may find application in the design of wound-healing patches with long-term activity., (Copyright © 2012 Elsevier B.V. All rights reserved.)

Aim: The use of nonsteroidal anti-inflammatory drugs (NSAIDs) in full-term pregnant women leads to fetal or neonatal toxicity, such as constriction of the ductus arteriosus (DA) and persistent pulmonary hypertension in the newborn. The aim of this study was to predict quantitatively the fetal toxicity of three NSAIDs (antipyrine, salicylic acid and diclofenac) using the transplacental pharmacokinetic parameters obtained from our previous placental perfusion studies., Methods: Human fetal plasma concentration profile after oral administration of each NSAID to the mother was estimated using the transplacental pharmacokinetic parameters and pharmacokinetic parameters in adult women. The fetal plasma concentration-response relationship for the three NSAIDs was estimated by pharmacokinetic/pharmacodynamic analysis of the results of previous studies investigating the effects of NSAIDs on the ratio of inner diameter of the DA to that of the pulmonary artery (DA/PA) in rats and the plasma concentration profiles of NSAIDs in pregnant rats., Results: The risk of constriction of the DA was well predicted by the model. Mean DA/PA ratio after oral administration of diclofenac to the mother was estimated to be 39.0%, whereas both of the corresponding values after oral administration of antipyrine and salicylic acid were estimated to be 5.9%. These results suggest that the fetal risk of diclofenac is higher than those of salicylic acid and antipyrine., Conclusions: This study presents a novel approach to predict quantitatively the fetal risk of NSAIDs administered to the mother. Human placental perfusion study and pharmacokinetic/pharmacodynamic analysis may provide basic data for predicting human fetal toxicity of drugs., (© 2011 The Authors. British Journal of Clinical Pharmacology © 2011 The British Pharmacological Society.)

Background: Ethylenediamine-N, N'-bis(2-hydroxyphenylacetic acid (EDDHA) is one of the most efficient iron-chelating agents employed to relieve iron chlorosis in plants. It has been well known for decades that this compound is photosensitive, but in spite of this fact its degradation pathways are virtually unknown. The aim of this work was to evaluate how the length of sunlight exposure and the concentration of irradiated EDDHA/Fe(3+) solutions influence the photostability of the chelate at constant pH. Moreover, the possible toxic effect of the chelate photodegradation products, elsewhere proposed, on soybean growth has been tested., Results: The photodecomposition of the chelate increased as the time of sunlight exposure increased, and resulted in a partial decomposition of the organic ligand. Moreover, EDDHA/Fe(3+) photodecomposition was highly correlated with the concentration of solution exposed. Plants did not present differences in recovery from chlorosis among treatments with and without decomposition products., Conclusions: EDDHA/Fe(3+) undergoes photodegradation, like other aminopolycarboxylic acids, being more degraded as solution concentration decreases and exposure time increases. The photodecomposition products salicylic acid, salicylaldehide and Salicylaldehyde ethylenediamine diimine tested did not have negative effects on soybean growth, at least in the short-term hydroponic experimental design tested., (Copyright © 2011 Society of Chemical Industry.)

More than 50 new inhibitors of the oncogenic Stat3 protein were identified through a structure-activity relationship (SAR) study based on the previously identified inhibitor S3I-201 (IC₅₀ =86 μM, K(i) >300 μM). A key structural feature of these inhibitors is a salicylic acid moiety, which, by acting as a phosphotyrosine mimetic, is believed to facilitate binding to the Stat3 SH2 domain. Several of the analogues exhibit higher potency than the lead compound in inhibiting Stat3 DNA binding activity, with an in vitro IC₅₀ range of 18.7-51.9 μM, and disruption of Stat3-pTyr peptide interactions with K(i) values in the 15.5-41 μM range. One agent in particular exhibited potent inhibition of Stat3 phosphorylation in both breast and multiple myeloma tumor cells, suppressed the expression of Stat3 target genes, and induced antitumor effects in tumor cells harboring activated Stat3 protein., (Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

The neuronal mechanism underlying the phantom auditory perception of tinnitus remains elusive at present. For over 25 years, temporary tinnitus following acute salicylate intoxication in rats has been used as a model to understand how a phantom sound can be generated. Behavioral studies have indicated that the pitch of salicylate-induced tinnitus in the rat is approximately 16 kHz. In order to better understand the origin of the tinnitus pitch measurements were made at the levels of auditory input and output; both cochlear and cortical physiological recordings were performed in ketamine/xylazine anesthetized rats. Both compound action potentials and distortion product otoacoustic emission measurements revealed a salicylate-induced band-pass-like cochlear deficit in which the reduction of cochlear input was least at 16 kHz and significantly greater at high and low frequencies. In a separate group of rats, frequency receptive fields of primary auditory cortex neurons were tracked using multichannel microelectrodes before and after systemic salicylate treatment. Tracking frequency receptive fields following salicylate revealed a population of neurons that shifted their frequency of maximum sensitivity (i.e. characteristic frequency) towards the tinnitus frequency region of the tonotopic axis (∼16 kHz). The data presented here supports the hypothesis that salicylate-induced tinnitus results from an expanded cortical representation of the tinnitus pitch determined by an altered profile of input from the cochlea. Moreover, the pliability of cortical frequency receptive fields during salicylate-induced tinnitus is likely due to salicylate's direct action on intracortical inhibitory networks. Such a disproportionate representation of middle frequencies in the auditory cortex following salicylate may result in a finer analysis of signals within this region which may pathologically enhance the functional importance of spurious neuronal activity concentrated at tinnitus frequencies., (Copyright © 2011 IBRO. Published by Elsevier Ltd. All rights reserved.)

Salicylic acid is a widely used nonsteroidal anti-inflammatory drug (NSAID). But it is known to cause serious liver damage occasionally. Mitochondrial dysfunction and oxidative stress are predicted to be the major factors of salicylic acid-induced liver injury. We investigated the influence of salicylic acid on ATP contents, oxygen consumption and lipid peroxidation in the presence of the same concentration of salicylic acid. Leakage of lactate dehydrogenase (LDH) was significantly higher in the presence of 5mM salicylic acid than in its absence. Salicylic acid-induced thiobarbituric acid-reactive substance (TBARS) formation and spontaneous chemiluminescence (CL) in rat hepatocytes, whereas antioxidants, such as promethazine (PMZ) and N,N-diphenylphenylenediamine (DPPD), suppressed both TBARS formation and LDH leakage. TBARS formation in rat liver microsomes was suppressed by diethyldithiocarbamate (a specific inhibitor of cytochrome P450 (CYP)2E1) and diclofenac (a specific inhibitor of CYP2C11). These results suggest that salicylic acid-induced lipid peroxidation was related to oxidative metabolism mediated by CYP2E1 and CYP2C11. On the other hand, 5mM salicylic acid induced a drastic decrease of ATP contents in rat isolated hepatocytes. Furthermore, mitochondrial respiration control ratio (RC ratio), calculated by State 3/State 4 also decreased with the increase of salicylic acid concentration. These findings suggest that salicylic acid would trigger mitochondrial dysfunction and cause ATP decrease, leading to lethal liver cell injury by lipid peroxidation, although this hypothesis remains to be elucidated in vivo., (Copyright (c) 2009 Elsevier Ireland Ltd. All rights reserved.)

Objectives/hypothesis: In gerbils in which tinnitus was induced by salicylate, neurons in the dorsal raphe nucleus were activated. Since about half the neurons in this region are serotonergic, this indicates that serotonin (5-HT) might play a role in the mechanisms of central tinnitus. The goal of this study was to determine if serotonergic neurons are activated during salicylate-induced tinnitus. Further, to determine if the same neurons might modulate the cochlea during tinnitus, neuroanatomical tract-tracing with 5-HT immunohistochemistry was used to determine if serotonergic neurons project to the gerbil cochlea., Study Design: Randomized, prospective, gerbils., Methods: Six gerbils were injected with salicylate (saline for controls). Four hours later, the gerbils were euthanized, perfused, and their brains collected for immunohistochemical labeling of 5-HT and c-fos. For the tract-tracing, FluoroGold was injected into the cochleae of 3 gerbils. The gerbils were euthanized and perfused 4-11 days later and the brains immunohistochemically processed for 5-HT., Results: More serotonergic neurons expressed c-fos in the salicylate-injected animals compared to controls. The increase was significant for 3 of the 8 major serotonergic cell groups including B7, B9, and the caudal linear nucleus. Despite robust labeling of olivocochlear and vestibular efferents with FluoroGold, 5-HT-labeled neurons containing FluoroGold were lacking., Conclusions: Salicylate-induced tinnitus activates serotonergic neurons in rostral cell groups. Activation of these neurons is not likely to influence cochlear function directly, but is likely to influence a number of auditory and non-auditory regions known to be involved with tinnitus.

The present study was designed to assess whether, in the presence of a depression of the cochlear amplifier i.e. a sensorineural hearing loss (SNHL), the inner hair cells (IHCs) require the presence of a normal endocochlear potential for transduction. An SNHL was induced by injecting salicylic acid (which binds to the motor protein prestin in the outer hair cells), and then furosemide (which depresses the endocochlear potential) was injected. Furosemide did not cause an additional elevation of the threshold of the auditory nerve brainstem evoked response (ABR) over that induced by the salicylic acid injection. Exposure to noise was also used to induce a SNHL in other mice, and then furosemide was injected. Here too furosemide did not cause an additional ABR threshold elevation over that induced by the noise. These results show that the IHCs (and the auditory nerve) can be excited in the presence of a SNHL (i.e. without the cochlear amplifier) and in the absence of an endocochlear potential. Possible mechanisms of excitation in such a state are discussed.

The inducibility of cytochrome P450 monooxygenases (P450s) and other xenobiotic-metabolizing enzymes is thought to reflect material and energy costs of biosynthesis. Efforts to detect such costs of detoxification enzyme induction, however, have had mixed success. Although they are rarely considered, ecological costs of induction may be a more significant evolutionary constraint on herbivores than material and energy costs. Because some P450-mediated metabolic transformations are bioactivation reactions that increase, rather than reduce, toxicity, maintaining high levels of P450 activity places an organism at risk of greater mortality in the presence of compounds that are bioactivated. We show that P450 inducibility in the generalist moth Helicoverpa zea in response to plant signaling substances, an adaptive response in a ditrophic interaction between herbivore and plant, becomes detrimental in the presence of a third trophic association with a plant pathogen that produces aflatoxin, a toxin that can be bioactivated by P450s. Consumption of plant signaling molecules, such as methyl jasmonate (MeJA) and salicylic acid (SA) enhanced the toxicity of aflatoxin B1 (AFB1) to H. zea that resulted in substantially more damage to larval growth and development. Among the P450 transcripts already cloned from this organism, two in the CYP6B and CYP321A subfamilies are shown to be induced in response to MeJA and SA, suggesting that they may mediate some of the observed bioactivations.

Xenobiotic diacylglycerols (DG) may induce pathological disorders by causing abnormal chromosomal segregation, which could be aneuploid. In this study, seven xenobiotic-diacylglycerols (four of drug origin and three of pesticide origin) were evaluated for their ability to induce aneuploidy in mammalian cultures using in vitro cytokinesis blocked micronucleus (CBMN) assay coupled with kinetochore labeling and interphase fluorescent in situ hybridization. Out of seven xeno-DGs, two (ibuprofen-DG and fenbufen-DG) induced statistically significant (P < 0.001) and dose-dependent increase in micronucleus induction, but this apparent micronucleus induction was very weak in case of fenbufen-DG. These MN were produced predominantly by aneugenic and clastogenic mechanisms, respectively, confirmed by immunofluorescent labeling of kinetochores. Fluorescent in situ hybridization analysis revealed that ibuprofen-DG induced significantly higher nondisjunction for chromosomes 10, 17, and 18. Other xenobiotic diacylglycerols (indomethacin-DG, salicylic acid-DG, 4-(2-methyl-4-chlorophenoxy) butanoic acid-DG (MCPB-DG), 2-(2-methyl-4-chlorophenoxy) propanoic acid-DG (MCPP-DG) and 2-(4-dichlorophenoxy)-butanoic acid-DG (2,4 DB-DG) did not induce micronuclei, but the concentrations tested did not reach levels that caused the marked growth suppression typically required for testing for regulatory testing purposes. However, the levels of growth suppression achieved were similar to that seen with ibuprofen-DG, which was positive. This study shows that xeno-DGs, which have been neglected in the past for their possible link to any pathological disorders, need serious assessment of their mutagenic potential.

Salicylate is a well-known substance to produce reversible tinnitus in animals and humans as well. It has been shown that systemic application of salicylate changes the neuronal spontaneous activity in several parts of the auditory pathway. The effects observed in central auditory structures in vivo could be based upon the changed afferent cochlear input to the central auditory system or in addition by a direct action of salicylate onto neurons within the auditory pathway. A direct influence of local salicylate application on spontaneous activity of central auditory neurons has already been described for the inferior colliculus (IC) in brain slice preparations. As spontaneous activity within all key structures of the central auditory pathway could play an important role in tinnitus generation, the present study investigated direct effects of salicylate superfusion on the spontaneous activity of the deafferented cochlear nucleus (CN), medial geniculate body (MGB), and auditory cortex (AC) in brain slices. Out of 72 neurons, 73.4% responded statistically significantly to the superfusate by changing their firing rates. 48.4% of them increased and 51.6% decreased their firing rates, respectively. The mean change of firing rate upon salicylate superfusion was 24.4%. All responses were not significantly different between the brain areas. The amount of neurons which responded to salicylate and the mean change of firing rate was much higher in the IC than in the CN, MGB and AC. This contributes to the hypothesis that salicylate-induced tinnitus is a phantom auditory perception mainly related to hyperexcitability of IC neurons. However, the present results suggest that the individual, specific salicylate sensitivity of CN, MGB and AC neurons can modulate the salicylate-induced generation of tinnitus.

Acid-sensing ion channel 3 (ASIC3) is the most sensitive acid sensor in sensory neurons that innervate into skin, muscle, heart, and visceral tissues. ASIC3 is involved in ischemia sensing, nociception, mechanosensation, and hearing, but how ASIC3-expressing neurons differ in their firing properties is still unknown. We hypothesized that ASIC3-expressing neurons have specialized firing properties, which, coupled with the heterogeneity of acid-sensing properties, accounts for various physiological roles. Here, we successfully identified ASIC3-expressing lumbar dorsal root ganglion (DRG) neurons whose transient proton-gated currents were blocked by salicylic acid (SA). The salicylic acid-sensitive (SAS) neurons did not exist in DRG neurons of mice lacking ASIC3. SAS neurons expressed distinct electrophysiological properties as compared with other DRG neurons. Especially, SAS neurons fired action potentials (APs) with large overshoot and long afterhyperpolarization duration, which suggests that they belong to nociceptors. SAS neurons also exhibited multiple nociceptor markers such as capsaicin response (38%), action potential (AP) with inflection (35%), or tetrodotoxin resistance (31%). Only in SAS neurons but not other DRG neurons was afterhyperpolarization duration correlated with resting membrane potential and AP duration. Our studies reveal a unique feature of ASIC3-expressing DRG neurons and a basis for their heterogeneous functions.