Defense Responses, Induced by p-Coumaric Acid and Methyl p-Coumarate, of Jujube ( Ziziphus jujuba Mill.) Fruit against Black Spot Rot Caused by Alternaria alternata.

The esterified fraction of jujube ( Ziziphus jujuba Mill.) peel extract showed strong antifungal activity on Alternaria alternata. p-Coumaric acid (pCA) was found to be the most predominant individual phenolic acid that was correlated highly with the antifungal activity of the esterified fraction. Thus, effects of postharvest treatments with pCA and its simplest esterified derivative methyl p-coumarate (MeCA) against black spot rot on jujube fruit caused by A. alternata were investigated. pCA and MeCA strongly suppressed in vitro growth of the fungus and significantly reduced postharvest Alternaria rot on fresh jujubes. Biochemical and transcriptional analysis revealed that pCA and MeCA regulated the expression of some genes encoding antioxidant enzymes and their enzymatic activities, enhanced the phenylpropanoid pathway metabolism, and activated the expression of genes encoding pathogenesis-related proteins. These results suggested that, apart from its direct antifungal activity, pCA and MeCA induced defense responses in jujube fruit against postharvest Alternaria rot.

In vitro immunocompetence of two compounds isolated from Polygala tenuifolia and development of resistance against grass carp reovirus (GCRV) and Dactylogyrus intermedius in respective host.

The present study was undertaken to isolate some compounds from methanol extract of Polygala tenuifolia and evaluate their immunostimulatory properties and antiviral activity using grass carp Ctenopharyngodon idella kidney (CIK) cells and GCRV. By applying insecticidal bioassay-guided, chromatography techniques and successive recrystallization, two purified compounds were obtained. The changes of expression of selected immune genes (Mx1, IL-1ß, TNFα, MyD88 and IgM) in C. idella kidney cell lines were evaluated after exposure to these isolated compounds. The results showed that compound 1 and 2 up-regulated to varying degrees of Mx1, IL-1ß, TNFα, and MyD88 in C. idella kidney cells. WST-8 kit assay verified the two compounds has no toxic effects on CIK cell, and furthermore, have in vitro antivirus activity. Especially, that there is keeping 79% cell viability when exposure to compound 2 (100 mg L(-1)). According to in vivo insecticidal assays against Dactylogyrus intermedius, compound 2 exhibited higher efficacy than compound 1, which was found to be 87.2% effective at the concentrations of 5 mg L(-1) and safe to goldfish (Carassius auratus). Besides, the purified compounds were identified by spectral data as: (1) 1,5-Anhydro-D-glucitol and (2) 3,4,5-trimethoxy cinnamic acid. Overall, the results indicate that bath administration of these compounds modulates the immune related genes in C. idella kidney cells and to some extent, eliminate the virus and parasitic infections.

Cinnamyl alcohol oxidizes rapidly upon air exposure.

BACKGROUND: Cinnamyl alcohol and cinnamal are frequent fragrance contact allergens. Both are included in the European baseline fragrance mix I, which is used for screening of contact allergy in dermatitis patients. OBJECTIVES: The aim of this study was to investigate the autoxidation of cinnamyl alcohol and to identify the oxidation products formed on air exposure. We also wanted to evaluate the effect of autoxidation on the sensitization potency of cinnamyl alcohol. METHODS: Samples of commercially available cinnamyl alcohol with and without purification were exposed to air, and the autoxidation was followed by chemical analysis. The analysis was performed with mass spectrometry (LC/MS/MS). Sensitization potencies of compounds were determined with the murine local lymph node assay (LLNA) in mice. RESULTS: Chemical analysis showed that the concentration of cinnamyl alcohol in the air-exposed samples decreased rapidly over time, and that autoxidation products were formed. Cinnamal, epoxy cinnamyl alcohol and cinnamic acid were identified as oxidation products. According to our study, cinnamal and epoxy cinnamyl alcohol were the first autoxidation products formed. The epoxy cinnamyl alcohol was shown to be the oxidation product with the highest sensitization potency. The analysis of our samples of commercially available cinnamyl alcohol showed that there was already a content of 1.5% cinnamal at the start of the autoxidation experiments. CONCLUSION: Cinnamyl alcohol readily autoxidizes upon air exposure, and forms strong sensitizers as determined by the LLNA. Cinnamal was formed in the largest amounts, showing that cinnamal is not only formed via bioactivation, as has previously been shown. A highly sensitizing epoxide was also identified and quantified in the oxidation mixture.

Propolis specimens from different locations of central Italy: chemical profiling and gas chromatography-mass spectrometry (GC-MS) quantitative analysis of the allergenic esters benzyl cinnamate and benzyl salicylate.

Propolis is a beehive product popular in natural medicine thanks to its noteworthy properties. Propolis is non-toxic but is responsible for allergic reactions in sensitive individuals. In this paper, we propose a new gas chromatography-mass spectrometry (GC-MS) analytical methodology for the quantitative analysis of two allergenic esters in propolis specimens, benzyl salicylate and benzyl cinnamate, and test it on specimens from different locations of central Italy. We also present the results obtained in the chemical characterization of the same specimens. The characterization showed that the resin fractions of all of the specimens are of poplar origin.

From xenobiotic chemistry and metabolism to better prediction and risk assessment of skin allergy.

Allergic contact dermatitis (ACD) is a condition that can have a serious impact on quality of life. The manifestation of ACD is dependent upon the primary sensitisation of an individual to a specific substance following skin exposure. It is important to identify and manage the risks associated with exposure to known skin sensitisers, in both the manufacture and use of consumer products. At present, the only validated approaches to conclusively identify sensitisation hazard and estimate potency are in vivo models such as the local lymph node assay. No in vitro test methods exist for this endpoint. There is an urgent need to develop novel in vitro/in silico testing or risk assessment strategies to replace animal testing. It is envisaged that such novel approaches can only be developed on the foundation of a good mechanistic understanding of skin sensitisation. Early stages of sensitisation are thought to be dependent upon the extent of compound absorption and bioavailability, rates of metabolic activation or detoxification and intrinsic reactivity of the bioavailable xenobiotic electrophile with skin protein nucleophiles. This review explores general chemical and metabolic aspects in relation to the potential formation of protein-hapten conjugates. Despite the complexities and poor understanding of some of the metabolic processes involved in skin sensitisation, it is possible to describe some of the relationships between chemical structures and the ability to form covalent conjugates with proteins. A prototypical group of xenobiotics that have been used to explore sensitisation mechanisms in some detail are selected cinnamic derivatives: a discussion of recent work using these compounds is presented as a case study. Novel aspects for future research in this area are also discussed.

Characterization of T cell responses to fragrances.

Fragrances are worldwide a major cause of allergic contact dermatitis (ACD), a delayed-type hypersensitivity reaction mediated by T lymphocytes. We investigated T cell responses to fragrances using peripheral blood mononuclear cells (PBMC) and T cells from skin lesions of fragrance-allergic patients. The components of a fragrance mixture (eugenol, isoeugenol, geraniol, oak moss, alpha-amyl cinnamic aldehyde, cinnamic aldehyde, cinnamic alcohol, and hydroxycitronellal) that is commonly used in the patch test were studied in vitro in the lymphocyte transformation test (LTT). PBMC from fragrance-allergic patients (n = 32) showed significant stimulations to all eight fragrances. The calculated stimulation indices (SI) varied between 2.1 and 21.8. The influence of metabolic enzymes on T cell stimulation was studied for two fragrances. Interestingly, stimulation of eugenol and isoeugenol was increased in the presence of antigen-modified human liver microsomes (CYP450) or recombinant CYP1A1 in five of seven cases. Furthermore, we established 18 T cell clones (TCC) from a skin lesion reacting specifically to eugenol. FACS analysis revealed that the majority (n = 15, 83%) of TCC were CD3(+), CD4(+), and HLA-DR(+). Seventeen percent (n = 3) of the clones were CD8(+). TCC (n = 4) released significant amounts of IL-2 and IFN-gamma but no IL-4 and IL-5. In addition, CD4(+) TCC (n = 3) showed antigen-induced cytotoxic activities against autologous B cells. In summary, we demonstrated for the first time that fragrance-specific CD4(+) and CD8(+) T lymphocytes are present in fragrance-allergic individuals. In addition, our results suggest that CYPs can be involved in the formation of the nominative antigen.

Mechanism of allergic cross-reactions--I. Multispecific binding of ligands to a mouse monoclonal anti-DNP IgE antibody.

A recently developed solid-phase binding assay was used to investigate the specificity of ligand binding to a mouse monoclonal anti-dinitrophenyl IgE [IgE(aDNP)]. All DNP-amino acids, that were tested, inhibited the binding of radio-labeled IgE(aDNP) to DNP covalently attached to polystyrene microtiter plates; however, the concentration for 50% inhibition varied within four orders of magnitude, DNP-L-serine being the most, DNP-proline the least potent inhibitor. In addition to DNP analogues a large number (2074) of drugs and other compounds were tested for their ability to compete with DNP for the binding site of IgE(aDNP). At the concentrations used for screening 59% of the compounds had no significant inhibition; 19% inhibited the binding of IgE(aDNP) more than 50%. Several families of compounds (tetracyclines, polymyxines, phenotiazines, salicylates and quinones) of effective competitors were found. Within these families change in the functional groups attached to the "family stem" had major effects on the affinity of ligand binding. The occurrence frequencies of interactions of ligands with IgE(aDNP) is in good agreement with a semi-empirical model for multispecific antibody-ligand interactions.

Cross-sensitization patterns in guinea pigs between cinnamaldehyde, cinnamyl alcohol and cinnamic acid.

Guinea pig maximization tests (GPMT) were performed with cinnamon substances. There was a certain degree of cross-reactivity between cinnamaldehyde, cinnamyl alcohol and cinnamic acid as animals sensitized to cinnamaldehyde reacted to the challenge with the three substances. Animals sensitized to cinnamyl alcohol reacted to cinnamyl alcohol and cinnamaldehyde, but not to cinnamic acid. Cinnamic acid did not sensitize guinea pigs. Compared to the challenge concentration for cinnamaldehyde, approximately a 15 times higher concentration of cinnamyl alcohol and a 25 times higher concentration of cinnamic acid were required to give positive reactions in animals sensitized to cinnamaldehyde. This could not be explained by differences in permeability properties, as the penetration profiles of the three substances through guinea pig skin in vitro showed permeability coefficients of the same order of magnitude under the test conditions. The study suggests that cinnamaldehyde is the "true" allergen, while cinnamyl alcohol and cinnamic acid are transformed in the skin to cinnamaldehyde, before contact allergic reactions can occur.

Potent contact allergen in the rubber plant guayule (Parthenium argentatum).

Chemical and dermatotoxicological investigations of the natural and processed resin of the Mexican rubber plant, guayule (Parthenium argentatum), has established the presence of a sesquiterpene cinnamic acid ester (guayulin A) that is a potent elicitor of allergic contact dermatitis in experimental animals. The guayule contact allergen is comparable to the poison ivy skin allergens as an elicitor of dermatitis in sensitized guinea pigs.

Studies on the cutaneous sensitization reaction of guinea pigs to purified aromatic chemicals.

The sensitization reactions to purified cinnamic aldehyde and its chemically related substances were investigated using the maximization test on guinea pigs. Mutual cross-sensitivities were also examined. Cinnamic aldehyde was found to be the most potent sensitizer in this series, and a strong cross-sensitizer. From the chemical structures of these substances, the following speculations were made. There was a tendency that as the number of hydrocarbons of alkyl groups replacing the alpha-hydrogen in the molecule increased, the rate of sensitization reaction declined. The conjugated system of double bonds was suspected to be a determinant factor in relation to the sensitization ability of the substances in this series.