Cosmetics with high water content are at a risk of being contaminated by micro-organisms that can alter the composition of the product or pose a health risk to the consumer. Pathogenic microorganisms such as Staphylococcus aureus and Pseudomonas aeruginosa are frequently found in contaminated cosmetics. In order to avoid contamination of cosmetics, the manufacturers add preservatives to their products. In the EU and the USA, cosmetics are under legislation and all preservatives must be safety evaluated by committees. There are several different preservatives available but the cosmetic market is dominated by a few preservatives: parabens, formaldehyde, formaldehyde releasers, and methylchloroisothiazolinone/methylisothiazolinone.Allergy to preservatives is one of the main reasons for contact eczema caused by cosmetics. Concentration of the same preservative in similar products varies greatly, and this may indicate that some cosmetic products are over preserved. As development and elicitation of contact allergy is dose dependent, over preservation of cosmetics potentially leads to increased incidences of contact allergy. Very few studies have investigated the antimicrobial efficiency of preservatives in cosmetics, but the results indicate that efficient preservation is obtainable with concentrations well below the maximum allowed.
Cell culture systems are widely used for the investigation of in vitro immunomodulatory effects of medicines and natural products. Since many pharmacological relevant compounds are water-insoluble, solvents are frequently used in cell based assays. Although many reports describe the cellular effects of solvents at high concentrations, only a few relate the effects of solvents used at low concentrations. In this report we investigate the interference of three commonly used solvents: Dimethyl sulfoxide (DMSO), ethanol and β-cyclodextrin with five different cell culture systems. The effects of the solvents are investigated in relation to the cellular production of interleukin (IL)-6 or reactive oxygen species (ROS) after lipopolysaccharide (LPS) stimulation. We show that DMSO above 1 % reduces readout parameters in all cell types but more interestingly the 0.25 and 0.5 % solutions induce inhibitory effects in some cell types and stimulatory effects in others. We also found that LPS induced ROS production was more affected than the IL-6 production in the presence of ethanol. Finally we showed that β-cyclodextrin at the investigated concentrations did not have any effect on the LPS induced IL-6 production and only minor effects on the ROS production. We conclude that the effects induced by solvents even at low concentrations are highly relevant for the interpretation of immunomodulatory effects evaluated in cell assays. Furthermore, these results show the importance of keeping solvent concentrations constant in serial dilution of any compound investigated in cell based assays.
Semliki Forest virus (SFV) utilizes a membrane fusion strategy to introduce its genome into the host cell. After binding to cell-surface receptors, virus particles are internalized through receptor-mediated endocytosis and directed to the endosomal cell compartment. Subsequently, triggered by the acid pH in the lumen of the endosomes, the viral envelope fuses with the endosomal membrane. As a result of this fusion reaction the viral RNA gains access to the cell cytosol. Low-pH-induced fusion of SFV, in model systems as well as in cells, has been demonstrated previously to be strictly dependent on the presence of cholesterol in the target membrane. In this paper, we show that fusion of SFV with cholesterol-containing liposomes depends on sphingomyelin (SM) or other sphingolipids in the target membrane, ceramide representing the sphingolipid minimally required for mediating the process. The action of the sphingolipid is confined to the actual fusion event, cholesterol being necessary and sufficient for low-pH-dependent binding of the virus to target membranes. The 3-hydroxyl group on the sphingosine backbone plays a key role in the SFV fusion reaction, since 3-deoxy-sphingomyelin does not support the process. This, and the remarkably low levels of sphingolipid required for half-maximal fusion (1-2 mol%), suggest that the sphingolipid does not play a structural role in SFV fusion, but rather acts as a cofactor, possibly through activation of the viral fusion protein. Domain formation between cholesterol and sphingolipid, although it may facilitate SFV fusion, is unlikely to play a crucial role in the process.
Occupational health symptoms related to bioaerosol exposure have been observed in a variety of working environments. Bioaerosols contain microorganisms and microbial components. The aim of this study was to estimate the total inflammatory potential (TIP) of bioaerosols using an in vitro assay based on granulocyte-like cells. A total of 129 bioaerosol samples were collected in the breathing zone of workers during their daily working routine at 22 biofuel plants. The samples were analyzed by traditional assays for dust, endotoxin, fungal spores, (133)--D-glucan, total number of bacteria, the enzyme N-acetyl--D-glucosaminidase (NAGase; primarily originating from fungi), Aspergillus fumigatus, and mesophilic and thermophilic actinomycetes; the samples were also assayed for TIP. In a multilinear regression four factors were significant for the TIP values obtained: endotoxin (P < 0.0001), fungal spores (P < 0.0001), (133)--D-glucan (P ؍ 0.0005), and mesophilic actinomycetes (P ؍ 0.0063). Using this model to estimate TIP values on the basis of microbial composition, the correlation to the measured values was r ؍ 0.91. When TIP values obtained in the granulocyte assay were related to the primary working area, we found that bioaerosol samples from personnel working in straw storage facilities showed high TIP values (Ϸ50 times the TIP of unstimulated controls). In contrast, bioaerosol samples from personnel with work functions in offices or laboratories showed low TIP values (Ϸ5 times the TIP of the unstimulated control). This indicates, as expected, that these areas were less contaminated. In conclusion, the granulocyte assay reacts to multiple contaminants in the environmental samples and can be used to obtain a measurement of TIP. Therefore, potential occupational health effects related to inflammation of the airways in a working environment can be estimated using this assay.For several years reports have related increased prevalence of respiratory symptoms to the exposure to bioaerosols containing, e.g., organic dust particles, actinomycetes, endotoxin, and fungal spores. Occupational health effects of bioaerosols have been reported in swine confinement houses and poultry farms as well as during hay handling. Airway diseases are frequent occupational disorders among farmers in many countries around the world (19,23). During mechanical handling, biofuels such as straw and wood chips release high amounts of various microbial components such as, e.g., actinomycetes, fungal spores, and endotoxin (11,25). Studies of personnel exposure to bioaerosols at biofuel plants reveal high concentrations of different microbial components and exposure levels higher than the suggested occupational exposure limits (10).Exposure limits or suggested exposure limits are usually based on studies where traditional microbial quantification methods have been applied and are normally related to the concentration of a single contaminant (e.g., endotoxin) (17) or to gravimetric measures of dust. Furthermore, it is recognized that exposur...
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