Growth of Staphylococcus aureus and synthesis of toxic shock syndrome toxin 1 in different in vitro systems.

Various complex and synthetic bacterial growth media have been used to study the growth of Staphylococcus aureus and the production of toxic shock syndrome toxin 1 (TSST-1) under certain in vitro culture conditions. Because of the biochemical and nutritional differences between these media and human menses, a program was designed to determine the growth and metabolism of S. aureus under conditions that more closely approximate in vivo conditions. Human menses, an artificial menses developed to match human menses, whole human blood, and complex bacterial culture media (with and without added whole human blood) were tested for the ability to support the growth of S. aureus and the production of TSST-1 in vitro. In addition, the impact of other organisms, commonly isolated from the human vagina, on the growth of S. aureus and the production of TSST-1 was evaluated. Results show that the environmental conditions provided by human menses are more closely approximated by the artificial menses and that neither commercial bacterial growth media alone nor complex media plus 25% or 50% human blood provide a menses-like environment for the growth of S. aureus and the production of TSST-1. Furthermore, the addition of a second organism to the S. aureus culture resulted in significant suppression of TSST-1 production, even when in vitro conditions were optimized for the production of TSST-1.

Response of various animal species to experimental infection with different strains of Staphylococcus aureus.

An experimental infection program was conducted in rabbits, pigs, and baboons with toxic shock syndrome (TSS)-associated and non-TSS-associated strains of Staphylococcus aureus to produce an animal model for TSS. TSS-associated strains of S. aureus--whether positive or negative for TSS toxin 1 (TSST-1)--could not be distinguished from non-TSS-associated strains of S. aureus by means of the rabbit whiffle-ball infection model; therefore, limited pilot infection studies were conducted in pigs and baboons. Experimental conditions were optimized in both the pig and the baboon studies to maximize the chance of producing TSS. Pigs infected with TSS-associated S. aureus strain CDC-11 developed some of the clinical signs observed in TSS (fever, hypotension, diarrhea, and vomiting). However, no changes were detected in clinical chemistry or hematology. Baboons infected with S. aureus strain CDC-11 showed only minimal signs of illness, i.e., lethargy, decreased food intake, and loose stools. TSS was not produced in pigs or baboons, even under optimal exposure conditions.

Cinnamic aldehyde: a survey of consumer patch-test sensitization.

The potential for cinnamic aldehyde, an important fragrance and flavour ingredient, to induce or to elicit delayed contact hypersensitivity reactions in man was evaluated by analysing patch-test data. Results of studies involving a total of 4117 patch tests on various consumer products and fragrance blends containing cinnamic aldehyde and on the material itself were collected from fragrance and formulator companies. The data indicate that cinnamic aldehyde contained in consumer products and fragrance blends at concentrations up to 6 X 10(-1)%, and patch-tested at concentrations up to 8 X 10(-3)%, has no detectable potential to induce hypersensitivity. Cinnamic aldehyde when tested alone induced a dose-related hypersensitivity response. According to published reports, cinnamic aldehyde elicited positive delayed hypersensitivity responses in dermatitic patients. However, results of the current survey show that when cinnamic aldehyde was tested alone or as part of a mixture in subjects in the general population, no pre-existing hypersensitivity reactions to the fragrance material were observed in any of the 4117 patch tests which constituted the survey. Cinnamic aldehyde at the concentrations contained in consumer products and fragrances, has a very low potential to induce hypersensitivity ('induced' reactions) or to elicit sensitization reactions ('elicited' reactions) in the general population.

Isoeugenol: a survey of consumer patch-test sensitization.

The potential of isoeugenol, an important fragrance and flavour ingredient, to induce delayed contact hypersensitivity or to elicit pre-existing sensitization reactions in man was evaluated by analysing patch-test data from dermatitic and non-dermatitic subjects. Results from a total of 6512 patch tests (involving approximately 5850 subjects) on isoeugenol alone and on various consumer products and fragrance blends containing isoeugenol, were collected from fragrance and formulator companies. Hypersensitivity induced by isoeugenol was concentration dependent. All but two of the reactions occurring in this survey were at exposure concentrations greater than or equal to 0.8% isoeugenol. No induced reactions occurred in the 1004 patch tests reported at isoeugenol concentrations between 0.03 and 0.5%. One induced reaction in 32 patch tests was attributable to isoeugenol at a concentration of 0.02% while another induced reaction in 23 patch tests conducted at the same concentration was resolved to an isoeugenol-eugenol mixture. One elicited reaction at an isoeugenol concentration of 0.04% occurred in the 6512 patch tests reported in this survey. This single elicitation was resolved to an isoeugenol-eugenol mixture, but the specific causative agent was not identified. The results of this survey indicate that isoeugenol has a very low potential for either eliciting pre-existing sensitization reactions ('elicited' reactions) or inducing hypersensitivity ('induced' reactions) in subjects exposed to consumer products containing this ingredient.

Eugenol and clove leaf oil: a survey of consumer patch-test sensitization.

The potential of eugenol and of clove leaf oil, which contains a high concentration of eugenol, to induce delayed skin hypersensitivity or to elicit reactions due to pre-existing skin sensitization in man was evaluated by analysing patch-test data. Results from a total of 11,632 patch tests on eugenol itself, on various consumer products containing eugenol and/or clove leaf oil, and on fragrance blends containing eugenol and/or clove leaf oil were collected from fragrance and formulation companies. One instance of induced hypersensitivity and one instance of pre-existing sensitization were observed at eugenol patch-test concentrations of 5 X 10(-2) and 9 X 10(-2)% respectively. In both cases, subsequent patch testing showed skin responses to mixtures of eugenol and isoeugenol, but no further attempt was made to define the causative agent. The survey indicates that, at the concentrations present in consumer products, eugenol alone or as part of clove leaf oil has a very low potential either to elicit pre-existing sensitization ('elicited' reactions) or to induce hypersensitivity ('induced' reactions).

Benzyl salicylate: a survey of consumer patch-test sensitization.

The potential of benzyl salicylate, an important fragrance and flavour ingredient, to induce hypersensitivity or to elicit reactions to pre-existing hypersensitivity in the general population was evaluated by analysing patch-test data. Results obtained from fragrance and formulator companies for a total of 10,538 patch tests on benzyl salicylate alone, on a variety of household and personal care consumer products and on fragrance blends containing benzyl salicylate were analysed as part of this survey. No induced or elicited responses directly attributable to benzyl salicylate were observed in the 35 patch tests on benzyl salicylate alone, or in the 10,503 patch tests on consumer products or fragrance blends containing benzyl salicylate. The highest concentration of benzyl salicylate tested in the consumer-product tests was 2 X 10(-1)%, and benzyl salicylate alone was tested at 10% in ethanol. This study indicates that benzyl salicylate has a very low potential to induce hypersensitivity ('induced' reactions) or to elicit reactions presumably attributable to pre-existing sensitization ('elicited' reactions) and thus supports the safe use of benzyl salicylate in consumer products and fragrance blends.

Simple and rapid gas-liquid chromatographic determination of diethylstilbestrol in biological specimens.

Diethylstilbestrol (DES) is readily extracted into toluene from biological fluids or tissue homogenates. Sodium carbonate is added to the initial mixture to eliminate potentially interfering substances. The toluene is extracted with a very small volume of aqueous trimethylphenylammonium hydroxide. This solution generates dimethyl DES in the vaporizer of a gas-liquid chromatograph. An internal standard, dienestrol (DI), is added at the beginning of the procedure and is partitioned and methylated in the same manner as DES. The DES and DI derivatives are well separated in less than 6 min on an ov-17 column. The entire analysis requires less than 15 min for a fluid specimen and less than 25 min for a solid tissue specimen. Seven samples can be analyzed each hour on a single column with a flame ionization detector. The relative standard deviations at levels from 2.5 to 100 ppm in bile are less than 5%. The lower limit of sensitivity is 8 ppb in a 1 ml bile sample.