Lise Moesby

Contamination versus preservation of cosmetics: a review on legislation, usage, infections, and contact allergy

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 micro‐organisms 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.

A comparative study of Mono Mac 6 cells, isolated mononuclear cells and Limulus amoebocyte lysate assay in pyrogen testing

Pyrogen induced secretion of interleukin 6 (IL-6) in Mono Mac 6 (MM6) cells was measured. The ability of the MM6 cell culture to detect pyrogens was compared to the Limulus amoebocyte lysate (LAL) test and isolated mononuclear cells (MNC). The detection limit of MM6 for lipopolysaccharide (LPS) and Staphylococcus aureus was comparable to that of MNC. Aspergillus niger and Candida albicans induced IL-6 in isolated MNC, but not in MM6. The detection limit for Salmonella typhimurium in the MM6 assay was comparable to that of the LAL assay. As expected, S. aureus and C. albicans did not show any LAL activity. A. niger and Influenza virus showed some activity in the LAL test, but could not be detected by MM6 cells. In conclusion, the MM6 assay is a good supplement to the current pyrogen assays for detection of LPS, S. aureus and S. typhimurium, but the MM6 assay could not detect A. niger, C. albicans and Influenza virus.

Isolation of immunomodulatory triterpene acids from a standardized rose hip powder ( Rosa canina L.)

A previously published systematic review and a metaanalysis have concluded that the consumption of standardized rose hip powder (Rosa canina L.) can reduce pain in osteoarthritis patients. Synovial inflammation has been suggested to play an important role in the pathogenesis of osteoarthritis and mainly to involve infiltration of the synovial membrane by macrophages. Therefore, the immunomodulatory effect of standardized rose hip powder of Rosa canina L. was investigated and active principles isolated using the Mono Mac 6 cell line as a model for human macrophages. Treatment of Mono Mac 6 cells with the residue of a crude dichloromethane extract of rose hip powder significantly and concentration dependently inhibited the lipopolysaccharide induced interleukin‐6 release. Through bioassay‐guided fractionation the immunomodulatory effect of the dichloromethane extract was correlated to a mixture of three triterpene acids; oleanolic acid, betulinic acid and ursolic acid (IC50 21 ± 6 µm). Further studies revealed that only oleanolic acid and ursolic acid, but not betulinic acid, could inhibit the lipopolysaccharide induced interleukin‐6 release from Mono Mac 6 cells when tested separately. Combination of either oleanolic acid or ursolic acid with betulinic acid enhanced the immunomodulatory effect of the two triterpene acids. Copyright

Low-level efficacy of cosmetic preservatives

Preservation using combinations of preservatives has several advantages. This study shows that the concentration of some of the most frequently used allergenic preservatives can be markedly lowered when they are combined with phenoxyethanol. The antimicrobial efficacy of cosmetic preservatives and known allergens of various potency [diazolidinyl urea, methylchloroisothiazolinone/methylisothiazolinone (MCI/MI), methylisothiazolinone (MI) and phenoxyethanol] was tested alone and in various combinations of two or three preservatives together. The preservatives were tested for minimum inhibitory concentration (MIC) values and possible synergy using fractional inhibitory concentration. MCI/MI was the only preservative showing low‐level MIC against all four tested microorganisms: Staphylococcus aureus, Pseudomonas aeruginosa, Candida albicans and Aspergillus niger. Different combinations of the preservatives indicated additive effects against the microorganisms. No combination of preservatives showed any inhibitory action on each other. Challenge tests with different concentrations and combinations were performed in a cosmetic cream. Diazolidinyl urea and MCI/MI alone were ineffective against C. albicans in a challenge test at concentrations up to 16 times higher than the observed MIC values. When combining phenoxyethanol with either one of the allergenic preservatives diazolidinyl urea, MCI/MI or MI, the cosmetic cream was adequately preserved at concentrations well below the preservatives’ MIC values as well as 10–20 times below the maximum permitted concentrations. By using combinations of preservatives, effective preservation can be achieved with lower concentrations of allergenic preservatives.

Assessment of the Total Inflammatory Potential of Bioaerosols by Using a Granulocyte Assay

ABSTRACT 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, (1→3)-β-d-glucan, total number of bacteria, the enzyme N-acetyl-β-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), (1→3)-β-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 (≈50 times the TIP of unstimulated controls). In contrast, bioaerosol samples from personnel with work functions in offices or laboratories showed low TIP values (≈5 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.

Adrenaline influences the release of interleukin-6 from murine pituicytes: role of β2-adrenoceptors

In this study, we examined the effect of adrenaline and interleukin-1beta on interleukin-6 secretion from cultured murine neurohypophyseal cells. Cells were cultured from neurohypophyses of 3- to 5-week-old mice and experiments were performed after 13 days in culture. Interleukin-6 was measured in 24-h samples using a sandwich fluoroimmunoassay. Unstimulated cells released 19+/-3 fmol interleukin-6/neurohypophysis/24 h (mean +/- S.E.M., n = 42). Adrenaline and interleukin-1beta increased the release of interleukin-6 from the cells in a concentration-dependent manner. Incubation with adrenaline (10(-6) M) or interleukin-1beta (11 pM) induced maximal secretion of interleukin-6, resulting in a 2.2-fold and 19.8-fold increase, respectively (P<0.01). The action of adrenaline (10(-6) M) and interleukin-1beta (1.1 pM) was examined separately and together. The sum of the effect of the two compounds given alone was significantly less (P<0.05) than the effect when adrenaline and interleukin-1beta were given together. We examined the effect of the beta-adrenoceptor antagonist propranolol (3.4x10(-6) M), the beta2-adrenoceptor antagonist (+/-)-1-[2,3-(Dihydro-7-methyl-1H-inden-4-yl)oxy]-3-[(1-methyl-eth yl)amino]-2-butanol (ICI 118551) (10(-7) M) and the beta1-adrenoceptor antagonist atenolol (10(-7) M and 10(-6) M) on the adrenaline-stimulated release of interleukin-6. Propranolol and ICI 118551 completely blocked the action of adrenaline, whereas atenolol was inactive. It is concluded that the stimulatory effect of adrenaline is mediated via beta2-adrenoceptors.

Pyrogen testing of lipid-based TPN using Mono Mac 6 monocyte cell line and DELFIA

Objective: Measurement of lipopolysaccharide (LPS) induced interleukin‐6 (IL‐6) secretion in Mono Mac 6 cells.

Endospores of B. subtilis are pyrogenic and activate Mono Mac 6 cells: importance of the CD14 receptor

The monocytic cell line Mono Mac 6 is sensitive to pyrogens and interleukin-6 secretion is induced after exposure to pyrogens. The aim of this study is to examine the pyrogenic activity and the interleukin-6-inducing capacity of the Gram-positive B. subtilis bacteria, endospores and isolated cell wall components. Furthermore the involvement of CD14 in activation of interleukin-6 release is investigated. All test substances are pyrogenic in the rabbit pyrogen test. The test substance is incubated with monocytic cells (Mono Mac 6) for 24 h and the secreted interleukin-6 is determined in a sandwich immunoassay. B. subtilis bacteria and endospores induce interleukin-6 in a dose-dependent manner. Endospores are less potent than bacteria. Lipoteichoic acid (LTA) isolated from B. subtilis induces interleukin-6 in a dose-dependent manner, whereas muramyl dipeptide (MDP) is unable to induce interleukin-6. Lipopolysaccharides (LPS) dose-dependently induce interleukin-6 release, but the curve differs from that of LTA both in shape and offset. The interleukin-6 secretion induced by LPS, LTA and B. subtilis bacteria can be blocked by 73-85% by an antibody directed against CD14, whereas the antibody only blocks 25% of B. subtilis endospores-induced interleukin-6 release. The results might indicate that B. subtilis endospores use an additional pathway to CD14 to activate mononuclear cells.

Lipidated α-peptide/β-peptoid hybrids with potent anti-inflammatory activity.

In this study, we investigated, optimized, and characterized a novel subclass of host defense peptide (HDP) mimics based on α-peptide/β-peptoid hybrid oligomers with an alternating cationic/hydrophobic design with respect to their ability to modulate the pro-inflammatory response by human primary leukocytes upon exposure to bacterial components. Structure-activity studies revealed that certain lipidated α-peptide/β-peptoid hybrid oligomers possess anti-inflammatory activities in the submicromolar range with low cytotoxicity, and that the anti-inflammatory activity of the HDP mimics is dependent on the length and position of the lipid element(s). The resulting lead compound, Pam-(Lys-βNSpe)6-NH2, blocks LPS-induced cytokine secretion with a potency comparable to that of polymyxin B. The mode of action of this HDP mimic appears not to involve direct LPS interaction since it, in contrast to polymyxin B, displayed only minor activity in the Limulus amebocyte lysate assay. Flow cytometry data showed specific interaction of a fluorophore-labeled lipidated α-peptide/β-peptoid hybrid with monocytes and granulocytes indicating a cellular target expressed by these leukocyte subsets.

Ultrasonication of pyrogenic microorganisms improves the detection of pyrogens in the Mono Mac 6 assay

The monocytic cell line Mono Mac 6 is sensitive to pyrogens. When exposed to pyrogens secretion of interleukin-6 is induced. However, some eukaryotic pyrogenic microorganisms are not detectable. The aim of this study is to introduce a pretreatment of samples to expand the detection range of the assay. The interleukin-6 inducing capacity of a broad spectrum of UV-killed and ultrasonicated microorganisms is examined in Mono Mac 6 cells. The interleukin-6 secretion is determined in a sandwich immunoassay (DELFIA). The Mono Mac 6 assay is able to detect UV-killed Bacillus subtilis, Staphylococcus aureus and Salmonella typhimurium, but neither Candida albicans nor Aspergillus niger. After ultrasonication of the microorganisms it is possible to detect C. albicans and A. niger. The interleukin-6 inducing ability of the examined microorganisms is in no case reduced after ultrasonic treatment. However, ultrasonication of S. aureus results in a 100-fold increase in the interleukin-6 response. Even after ultrasonication Streptococcus faecalis can not be detected. Ultrasonication is an easy and simple method for expanding the detection range in the Mono Mac 6 assay.