Neurobehavior and neuron damage following prolonged exposure of silver nanoparticles with/without polyvinylpyrrolidone coating in Caenorhabditis elegans.

Silver nanoparticles (AgNPs) have become widespread in the environment with increasing industrial applications. But the studies about their potential health risks are far from enough, especially in neurotoxic effects. This study aimed to investigate the neurotoxic effects of longer-term exposure (prolonged exposure for 48 h and chronic exposure for 6 days) of 20nm AgNPs with/without polyvinylpyrrolidone (PVP) coating at low concentrations (0.01-10 mg·L-1 ) to Caenorhabditis elegans. The results suggested that exposure to AgNPs induced damage to nematode survival, with the longest and relative average life span reduced. Exposure to AgNPs caused neurotoxicity on locomotion behaviors (head thrashes, body bends, pharyngeal pumping frequency, and defecation interval) and sensory perception behaviors (chemotaxis assay and thermotaxis assay), as well as impaired dopaminergic, GABAergic, and cholinergic neurons, except for glutamatergic, based on the alters fluorescence intensity, in a dose- and time-dependent manner. Further investigations suggested that the low-dose AgNPs (0.01-0.1 mg·L-1 ) exposure raises receptors of GABAergic and dopamine in C. elegans at the genetic level, whereas opposite results were observed at higher doses (1-10 mg·L-1 ), which implied that AgNPs could cause neurotoxicity by impairing neurotransmitter delivery. The PVP-AgNPs could cause a higher fatality rate and neurotoxicity at the same dose. Notably, AgNPs did not cause any deleterious effect on nematodes at the lowest dose of 0.01 mg·L-1 . In general, these results suggested that AgNPs possess the neurotoxic potential in C. elegans and provided useful information to understand the neurotoxicity of AgNPs, which would offer an inspiring perspective on the safe application.

Lippia origanoides essential oil: an efficient and safe alternative to preserve food, cosmetic and pharmaceutical products.

AIMS: The aim of this work was to evaluate the efficacy and safety of Lippia origanoides essential oil as a preservative in industrial products. METHODS AND RESULTS: The composition, antimicrobial activity, mutagenic and toxic potential of L. origanoides were determined. Then, the effect of essential oil as a preservative in food, cosmetics and pharmaceutical products was evaluated. The essential oil of L. origanoides consisted mainly of oxygenated monoterpenes (38·13%); 26·28% corresponded to the compound carvacrol. At concentrations ranging from 0·312 to 1·25 µl ml-1 and in association with polysorbate 80, the essential oil of L. origanoides inhibited the growth of all the tested micro-organisms. The medium lethal dose in mice was 3·5 g kg-1 , which categorizes it as nontoxic according to the European Union criteria, and negative results in the Ames test indicated that this oil was not mutagenic. In combination with polysorbate 80, the essential oil exerted preservative action on orange juice, cosmetic and pharmaceutical compositions, especially in the case of aqueous-based products. CONCLUSIONS: Lippia origanoides essential oil is an effective and safe preservative for orange juice, pharmaceutical and cosmetic products. SIGNIFICANCE AND IMPACT OF THE STUDY: This study allowed for the complete understanding of the antimicrobial action and toxicological potential of L. origanoides essential oil. These results facilitate the development of a preservative system based on L. origanoides essential oil.

Oral subchronic exposure to silver nanoparticles in rats.

Because of their extremely small size, silver nanoparticles (AgNPs) show unique physical and chemical properties, with specific biological effects, which make them particularly attractive for being used in a number of consumer applications. However, these properties also influence the potential toxicity of AgNPs. In this study, we assessed the potential toxic effects of an in vivo oral sub-chronic exposure to polyvinyl pyrrolidone coated AgNPs (PVP-AgNPs) in adult male rats. We also assessed if oral PVP-AgNPs exposure could alter the levels of various metals (Fe, Mg, Zn and Cu) in tissues. Rats were orally given 0, 50, 100 and 200 mg/kg/day of PVP-AgNPs. Silver (Ag) accumulation in tissues, Ag excretion, biochemical and hematological parameters, metal levels, as well as histopathological changes and subcellular distribution following PVP-AgNPs exposure, were also investigated. After 90 days of treatment, AgNPs were found within hepatic and ileum cells. The major tissue concentration of Ag was found in ileum of treated animals. However, all tissues of PVP-AgNPs-exposed animals showed increased levels of Ag in comparison with those of rats in the control group. No harmful effects in liver and kidney, as well as in biochemical markers were noted at any treatment dose. In addition, no hematological or histopathological changes were found in treated animals. However, significant differences in Cu and Zn levels were found in thymus and brain of PVP-AgNPs-treated rats.

Excipientes farmacêuticos: perspectivas dos polissacarídeos na pesquisa e desenvolvimento de novos sistemas de liberação / Pharmaceutical excipients: perspectives for polysaccharides in the research and development of new drug delivery systems

Excipientes em geral são conceituados como substâncias auxiliares diretamente envolvidas na composição das diversas formulações farmacêuticas. Constituintes de diferentes sistemas terapêuticos viabilizam o sucesso da forma farmacêutica final, apresentando características e propriedades diferentes daquelas pertinentes aos fármacos. Excipientes farmacêuticos constituem elementos de elevado destaque na formulação dos medicamentos, uma vez que, exercem efetivo papel na garantia de obtenção da forma farmacêutica adequada ao uso e ao efeito terapêutico desejado, regendo e influenciando de maneira significativa a cedência do princípio ativo contido no medicamento. Polissacarídeos são polímeros de monossacarídeos, encontrados abundantemente na natureza, a baixo custo, e devido as suas propriedades, de baixa toxicidade, biodegradabilidade, características filmogênicas, facilidade de derivatização, disponibilidade a baixo custo, têm constituído elementos de elevado interesse e destaque nas pesquisas farmacêuticas. Estes polímeros naturais encontram vasto campo de aplicação incluindo a melhoria de diversas propriedades das formulações de formas farmacêuticas sólidas tradicionais, sendo estas representadas pela facilidade de escoamento dos granulados, boa uniformidade de peso e otimização da dureza em comprimidos. Prioritariamente, encontram posição de destaque na pesquisa e desenvolvimento de novos sistemas para liberação modificada de fármacos quando aplicados nos sistemas reservatórios (revestimento farmacêutico), sistemas matriciais, ou como ôcarregadoresö macromoleculares e/ou biodegradáveis

Biotransformation of coumarin by rodent and human cytochromes P-450: metabolic basis of tissue-selective toxicity in olfactory mucosa of rats and mice.

Coumarin was previously found to cause tissue-selective toxicity in the olfactory mucosa (OM) of rats and mice, with rats being the more sensitive species. The aim of this study was to explore the role of target tissue biotransformation in OM-selective toxicity and the metabolic basis of the species differences in coumarin toxicity. At least six coumarin metabolites were detected in OM microsomal reactions, with o-hydroxyphenylacetaldehyde (o-HPA) being the most abundant. Formation of o-HPA was inhibited by reduced glutathione, confirming its origin from a reactive intermediate. There were significant differences in the rates and metabolite profiles of coumarin metabolism in the livers of Wistar rats and C57BL/6 mice. The rates of metabolic activation of coumarin, as indicated by the formation of o-HPA, were comparable in OM microsomes of the two species but about 25- and 3-fold higher in OM than in liver microsomes of rats and mice, respectively. Thus, target tissue activation seems to play an important role in the tissue-selective toxicity, whereas differences in the rates of hepatic metabolism may be responsible for the species difference in olfactory toxicity. Purified, heterologously expressed mouse CYP2A5 and CYP2G1 produced 7-hydroxycoumarin and o-HPA as the predominant products, respectively. Kinetic analysis and immunoinhibition studies indicated that the OM-specific CYP2G1 plays the major role in metabolic activation of coumarin. Furthermore, of 13 human cytochrome P-450s (P-450s) examined, five (CYP1A1, CYP1A2, CYP2B6, CYP2E1, and CYP3A4) were active in the metabolic activation of coumarin, suggesting a potential risk of coumarin toxicity in humans.

Residual monomer/additive release and variability in cytotoxicity of light-curing glass-ionomer cements and compomers.

In previous studies, light-cured glass-ionomer cements have been shown to evoke cytotoxic reactions. It was the purpose of this investigation (a) to determine the nature of the ingredients released into an aqueous medium from 2 light-cured glass-ionomer cements (GICs) and 3 compomers; (b) to evaluate the cytotoxicity of these extracts; and (c) to correlate the extent of the cytotoxic effects with eluted substances. Specimens of 2 light-cured GICs and 3 compomers were prepared and extracted in distilled water or cell culture medium for 24 hrs (surface-liquid ratio 42.4 mm2/mL). The aqueous eluates were analyzed by gas chromatography/mass spectrometry (GC/MS). The relative amounts of the components released from various products were compared by means of an internal caffeine standard [%CF]. For evaluation of cytotoxic effects, permanent 3T3 fibroblasts were incubated with medium extracts for 24 hrs. In addition, the ED50 concentration of the photoinitiator diphenyliodoniumchloride (DPICl) was determined. In all extracts, several water-elutable organic substances were found: (Co)monomers (especially HEMA and ethylene glycol compounds), additives (e.g., camphorquinone and diphenyliodoniumchloride), and decomposition products. The extracts of 3 products inhibited cell growth only moderately, whereas the light-cured GIC Vitrebond and the compomer Dyract Cem revealed severe cytotoxic effects. Vitrebond liberated the initiator DPICl, whereas Dyract Cem segregated a relatively high quantity [2966 %CF] of the comonomer TEGDMA in comparison with the other products. The present data show that TEGDMA and DPICl may be regarded as the prime causes for cytotoxic reactions evoked by the investigated light-cured glass-ionomer cements or compomers. Therefore, leaching of these substances should be minimized or prevented.

Effects of viscoelastics on bovine corneal endothelial cells in vitro.

PURPOSE: To evaluate the possible toxic effects of sodium hyaluronate and hydroxypropyl methylcellulose on corneal endothelium. METHODS: Cultured bovine corneal endothelial cells (BCEC) were treated with either original Healon (10 mg/ml) or Methocel (20 mg/ml) for 1 h, or with various dilutions of these substances in culture medium for up to one week. The toxicity of the viscoelastics was assessed in terms of lactate dehydrogenase (LDH) release into the supernatant and of cell density. RESULTS: Neither Healon nor Methocel in a dilution of 2 mg/ml enhanced LDH release after 72 h incubation, when compared with the control in a confluent model. In a proliferation model neither diluted Healon nor Methocel showed apparent inhibitory or stimulatory effects on the growth of BCEC up to the highest concentration we tested. When a BCEC monolayer was covered for 1 h with either undiluted Healon or undiluted Methocel, a significant, though transient, higher LDH release was induced. CONCLUSION: The results indicate that the diluted viscoelastics are safe for long time contact with BCEC, but undiluted they may temporarily interfere with the metabolism of the cytoplasm membranes of BCEC.

Effect of inhaled preservatives on asthmatic subjects. II. Benzalkonium chloride.

Benzalkonium chloride has been used as a preservative in some antiasthma respirator solutions and is known to cause bronchoconstriction in asthmatic subjects. To increase understanding of how it causes bronchoconstriction, the characteristics of airway response in 28 asthmatic subjects were documented. Subjects inhaled histamine, in doses ranging from 0.03 to 7.8 mumol, or benzalkonium in doses ranging from 0.04 to 5.33 mumol on separate days. The dose of histamine or benzalkonium that caused a 20% fall in the 1-s forced expiratory volume (PD20FEV1) was measured. All subjects responded to histamine, with PD20FEV1 values in the range of 0.14 to 7.8 mumol and 17 responded to benzalkonium, with PD20FEV1 values in the range 0.35 to 5.55 mumol. Subjects who responded to benzalkonium were more sensitive to histamine (mean PD20FEV1 0.44 mumol) than subjects who did not respond (mean PD20FEV1 1.84 mumol) and, among the benzalkonium responders, there was a significant correlation between PD20FEV1 values for histamine and benzalkonium (r = 0.5, p less than 0.05). Inhalation of benzalkonium enhanced subsequent responses to histamine, causing a decrease in mean PD20FEV1 from 0.51 to 0.18 mumol histamine (p less than 0.001), but did not alter subsequent responses to benzalkonium. The response to benzalkonium reached a maximum 1 min after inhalation and was slow to recover, taking up to 60 min to return to baseline values. Response to benzalkonium was inhibited by 8 mg cromolyn sodium but not by 160 micrograms ipratropium bromide. The characteristics of the response to benzalkonium suggest a mechanism of action via release of mediators.

Toxic effects of ophthalmic preservatives on cultured rabbit corneal epithelium.

We investigated the effects of the ophthalmic preservatives thimerosal and sorbic acid on the proliferation and survival of rabbit corneal epithelial cells in tissue culture. Normally, explants of corneal epithelium grow vigorously during the first 7 days in culture. With 0.004% thimerosal present in the culture medium, the normal proliferation of corneal cells is suppressed completely. When 0.1% sorbic acid is present, proliferation is delayed and the lifespan of the corneal cells is reduced. After a 1-h exposure to concentrations of thimerosal of 0.0005% or greater, virtually all corneal cells present in established cultures are killed. These results suggest that use of ophthalmic preparations containing these chemicals may affect the metabolic and proliferative capacity of the corneal epithelium adversely.

[Individual sensitivity of cilia. A simple in vitro test method]. / Individuelle Empfindlichkeit der Cilien. Eine einfache in-vitro-Testmethode.

A simple patient-related method is described that determines the effect of toxic substances on the ciliary activity of human nasal cilia cells. The method is suitable for investigating and numerically determining substance-specific toxicity and individual sensitivity even between patients of different states of health. This is illustrated by the example of two preservatives introduced in human medicine: chlorhexidine and merthiolate.

Propylene glycol-induced cholesteatoma in chinchilla middle ears.

Propylene glycol is a solvent commonly used in topical otic preparations. This study examines the occurrence of inflammatory changes and cholesteatoma in chinchilla middle ears after the application of propylene glycol in varying concentrations. A total of 32 ears were studied, divided into four treatment groups. Three groups received propylene glycol in concentrations of 10%, 50%, and 90%. One group received normal saline. Six weeks after the application of propylene glycol to the middle ear through the bulla, examination revealed cholesteatoma, tympanic membrane perforations, and middle ear adhesions in most of the ears subjected to 50% and 90% propylene glycol. Only one ear treated with 10% propylene glycol showed a cholesteatoma, while the group treated with normal saline showed only mild inflammation. Histologic preparations confirmed cholesteatoma and revealed replacement of the normal columnar epithelium by keratinizing stratified squamous epithelium, inflammatory infiltration, and granulation tissue eroding underlying bone. We conclude that exposure to propylene glycol in high concentrations will consistently produce cholesteatoma in chinchilla middle ears. Although the effects of propylene glycol in the human middle ear are yet to be investigated, we recommend the avoidance of otic preparations containing high concentrations of propylene glycol in patients with tympanic membrane perforations.

Kathon CG: a review.

Kathon CG, a cosmetics preservative containing, as active ingredients, 5-chloro-2-methyl-4-isothiazolin-3-one and 2-methyl-4-isothiazolin-3-one, appears to be a frequent cause of contact dermatitis in Europe. In the United States, where Kathon CG was introduced some 5 years later, the use of this preservative system for cosmetics and toiletries is rapidly increasing. Undoubtedly cases of contact sensitization will soon emerge in this country. Most cases of contact allergy are caused by the use of moisturizing creams on (slightly) damaged skin. Sensitization by the use of cosmetic products on previously healthy skin, especially the face, does occur but appears to be less frequent. Rinse-off products do not seem to have a substantial potential for the induction and elicitation of contact allergic reactions to Kathon CG because of dilution of the product and the allergen with water as well as a short contact time with the skin. This review provides a synopsis of current knowledge on the preservative system Kathon CG, with emphasis on the risk of sensitization and diagnostic procedures.

Sensitization studies in the guinea pig with the active ingredients of Euxyl K 400.

The preservative Euxyl K 400 consists of the 2 active ingredients, 2-phenoxyethanol and 1,2-dibromo-2,4-dicyanobutane. Sensitization studies with the guinea pig maximization test were performed with these substances, but no sensitizing capacity was demonstrated in the case of either compound.

Human safety study of body lotion containing Kathon CG.

The safety of Kathon CG biocide as a preservative in leave-on body lotions was assessed by 2 double-blind studies, using similar protocols. A total of 209 healthy male and female subjects aged 18 to 65 years, 100 in California (72 test subjects, 28 controls) and 109 in Florida (88 test subjects, 21 controls) completed the studies which included pre- and post-use phase diagnostic patch testing with Kathon CG 100 ppm active ingredient, and 13 weeks daily applications of either a test lotion containing Kathon CG 15 ppm active ingredient or a control lotion without Kathon CG. No evidence of irritation or sensitization attributable to use of the biocide was found during regular dermatological examinations during the use phase. Post-use phase patch testing produced negative results in all subjects with the exception of 1 control subject in Florida who had positive readings at the 2- and 4-week post-use phase patch testing. Overall, these studies show there is minimal, if any, risk of adverse effects associated with the use of Kathon CG 15 ppm active ingredient in a leave-on application.