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Propolis allergy (IV). Studies with further sensitizers from propolis and constituents common to propolis, poplar buds and balsam of Peru.

26 different compounds have been investigated experimentally for their sensitizing capacity in guinea pigs. 19 of these occur in propolis as well as in poplar bud exudates, and 14 of them are also found in balsam of Peru. 4 caffeates and benzyl isoferulate were found to be strong sensitizers. 7 compounds were moderate, and 13 compounds showed only weak sensitizing potency. Methyl cinnamate was negative. Patch tests in 11 propolis-sensitive patients once more revealed 3-methyl-2-butenyl caffeate and phenylethyl caffeate as the major sensitizers. In addition to the 8 compounds already known to occur in propolis as well as in balsam of Peru, we detected 5 further substances that both materials have in common. Among these, benzyl isoferulate is considered a noteworthy sensitizer. Coniferyl benzoate, which was shown to be a moderate sensitizer, is present in fresh samples of balsam of Peru, while in propolis it has been detected only once so far. The flavonoid aglycones occurring in poplar bud exudates, and hence also in propolis, are weak sensitizers which play only a minor role in propolis hypersensitivity.

Allergens

[On the antimicrobial activity of propolis and propolis constituents (author's transl)].

After a survey of the literature on the antimicrobial activity of the bee product propolis, the authors discuss their own findings as compared to the chemotherapeutical agents streptomycin, oxytetracycline, chloramphenicol, nystatin, griseofulvin and sulphamerazine. According to the results obtained by testing 25 isolated constituents on Bacillus subtilis, Staphylococcus aureus, Candida albicans and Trichophyton mentagrophytes, the antimicrobial properties of this mixture of natural substances are mainly attributable to the flavonoids pinocembrin, galangin, pinobanksin, pinobanksin-3-acetate as well as to the p-coumaric acid benzyl ester and a caffeic acid ester mixture. None of the isolated substances was as potent as the antibiotics tested for the purpose of comparison. The relatively good antimycetic activity of the 5,7-dihydroxyflavanone pinocembrin seems noteworthy. Finally, possible mechanisms of the antimicrobial action of the flavonoids are discussed.

Anti-Bacterial Agents

Biological properties and clinical application of propolis. I. Some physico-chemical properties of propolis.

The presence of 19 elements has been shown in the ethanol extracts of propolis (EEP). Three fractions have been obtained by filtration through a structural gel that did not show an initial antibacterial activity when investigated separately. Fractions 2 and 3 joined together have regained this activity. EEP solutions maintain their anitbacterial activity in acidic or neutral pH. Insensitivity of EEP solutions on temperautre of 75 degrees C for 30 min has been found.

Bees

Biological properties and clinical application of propolis. III. Investigation of the sensitivity of Staphylococci isolated from pathological cases to ethanol extract of propolis (EEP). Attempts on inducing resistance in laboratory Staphylococcus strain to EEP.

Staphylococci isolated from pathological material exhibited a reduced sensitivity to ethanol extract of propolis (EEP) in 90% of cases. No cross-resistance of the staphylococci to EEP and to any commonly used antibiotics was found. The induction of resistance to EEP in laboratory strain of Staphylococcus aureus (Oxford 209 P) can be achieved already after serial passages on nutrient media containing EEP. Culturing Staphylococcus resistant to EEP in an environment devoid of this compound caused a remission to sensitivity of the strain investigated.

Anti-Bacterial Agents

Biological properties and clinical application of propolis. IV. The action of ethanol extract of propolis (EEP) on cells cultured in vitro.

An increase of total cell number was shown in the cell culture in vitro under the influence of ethanol extract of propolis (EEP). Addition of EEP to the nutrient medium of the cells caused a strong activation of mitoses. Besides, distinctly intensified metabolism of these cells expressed by a strong activation of NADH2-reductase was also observed.

Cells, Cultured

Biological properties and clinical application of propolis. X. Preliminary pharmacological evaluation of ethanol extract of propolis (EEP).

A study consisting of an examination of the acute toxicity of ethanol extract of propolis (EEP) in mice, its effect on spontaneous movement in mice and rats, its analgesic properties and its influence on body temperature in mice was conducted. Also examined was the activity of EEP on animals under the influence of narcotics and spontaneous movement under the influence of amphetamine, its effects on blood pressure and respiration in rats. The results of these examinations indicate that EEP injected i.p. has a weak general effect on the experimental animals.

Amphetamine

[Selective effect of propolis in the isolation of Listeria monocytogenes (author's transl)].

Propolis is a substance produced by honeybees. It is inhibitory to some bacteria species, mainly Gram-positive bacteria, but less inhibitory to Listeria monocytogenes (L.m) than to the other Gram-positive bacteria tested. In order to obtain selective growth of L.m. from contaminated samples, the effect of propolis in plating media and broths on various strains of bacteria was examined. Table I shows the effect of increasing concentrations of propolis in tryptose-agar (TA). L.m. tolerated higher concentrations of propolis than Streptococcus viridans and Staphylococcus aureus. L.m. grew well in tryptosebroth (TB) that contained 0.15 mg propolis pr. ml medium, while Streptococcus viridans and Streptococcus agalactiae were completely inhibited as seen in Table II. Table III shows that when serum was added to the agar, the inhibitory effect was reduced. It can also be seen that Gram-negative bacteria grew quite well on media that contained 0.19 mg propolis pr. ml. To reduce the growth of Gram-negative bacteria, nalidixic acid was added to the medium. Table IV illustrates growth of various species of bacteria in tryptosephosphatebroth (TFB) with or without propolis and nalidixic acid. Most of the strains tested were inhibited, but Pseudomonas aeruginosa and to some extent faecal streptococci were able to grow in the medium that contained the selective substances. As a conclusion it seems that propolis may be a valuable additive to a medium for the selective isolation of L.m.

Anti-Bacterial Agents

Effects of propolis on dental caries in rats.

Propolis, the resinous hive product collected by bees, is important in the defense of the hive. The effects of propolis on growth and glucosyltransferase activity of Streptococcus sobrinus 6715, Streptococcus mutans PS14 and Streptococcus cricetus OMZ61 in vitro, and on dental caries in rats infected with S. sobrinus 6715 were investigated. Propolis had antimicrobial activity against S. sobrinus, S. mutans and S. cricetus, and inhibited both water-insoluble glucan synthesis and glucosyltransferase activity. In rats inoculated with S. sobrinus, about half of their fissures were carious, while dental caries was significantly less (p = 0.01) in rats given propolis. Dental caries was markedly decreased by the multiple actions of propolis which had antimicrobial activity, inhibited water-insoluble glucan synthesis, and inhibited glucosyltransferase activity. No toxic effects of propolis on the growth of rats were observed under experimental conditions in this study. These results suggest that propolis can control dental caries in the rat model system.

Animals

Contact dermatitis from propolis.

Two patients with contact dermatitis due to the natural product propolis (bee glue) are reported. They presented perioral eczema and stomatitis which were recalcitrant until propolis was considered as the cause. Patch tests with propolis preparations were positive in both patients, and, furthermore, in the second patient the lesions relapsed after provocation tests. European standard patch test including balsam of Peru were negative. The complexity of propolis, its supposed anti-inflammatory effect due to flavonoids, and the sensitizing agents originating mainly from the poplar trees are discussed together with the cross-sensitization to balsam of Peru. Contact dermatitis due to propolis should be considered in unexplained eczemas, mainly perioral but also in other areas, as propolis preparations are available also as ointments and cosmetic creams.

Aged

[Propolis allergy].

The incidence of propolis contact sensitivity was 1.2-3.3% among 7483 hospital patients during the period 1981-1987, while the prevalence among 1,558 healthy volunteers was 0.64%. A group of 26 patients with contact allergy to propolis was challenged with identified substances isolated from propolis. The mixture of 3-methyl-2-butenylester (3M2B) and 3-methyl-3-butenylester (3M3B) from caffeic acid caused a positive reaction in 7 patients, and in 5 of 9 patients there was a positive reaction to the mixture of 3M2B and 3M3B from diacetyl-caffeic acid. Among the 19 patients challenged with the flavonoid group, 3 had positive reactions, and individual derivatives of cinnamic acid caused positive reactions in 3 and 4 patients each. When five components taken from poplar buds were tested, the one that provoked the largest number of positive reactions was the methanol component (15 of 19 patients). The results show that propolis contact allergy is not caused by one main allergen, but by several allergens varying in chemical composition; the presence of these in propolis depends on the nature of the source plant and the place and time of collection by the bees.

Adult

Nutrikinetics and bioavailability of Promunel®, a standardized poplar-type propolis phenolic extract: a double-blinded, placebo-controlled, cross-over, randomized trial.

Brown poplar-type propolis has been recognized and used for centuries to help prevent upper respiratory tract infections (URTIs). However, the scarce, incomplete information in humans on the nutrikinetics and bioavailability of its phenolic constituents, combined with a lack of standardization in its phenolic content and profile pose major challenges to develop bioactive ingredients. Thus, the aim of this study was to establish the nutrikinetics and total bioavailability (NKBA) parameters of brown poplar-type propolis phenolics in humans using the Standardized Propolis Extract (SPE) Promunel®. To achieve this, a 48 h NBKA study was conducted following a double blinded, randomized, placebo-controlled, cross-over design in healthy humans (n = 10) with two doses of SPE (1X = 400 mg or 4X = 1600 mg). Phenolic compounds were detected, identified and quantified in the extract, plasma and urine through different LC-MS/UV technologies. The SPE used is a rich (304.44 ± 15.61 µmol mg-1) and diverse source of phenolic compounds (5 sub-families). A total of 63 and 85 phenolic metabolites were identified and quantified in plasma and urine, mostly in the form of glucuronides and sulfates. In plasma, phenolic metabolites reached Cmax (1.22 ± 0.20 for 1X and 4.80 ± 0.48 µM for 4X) after 1 h of SPE intake, while urinary excretion occurred mostly during the first 3 h after. The total net bioavailability of SPE phenolic compounds at 48 h was 57.16 ± 5.71% for 1X and 43.82 ± 6.77% for 4X. Generally, the data between SPE 1X and 4X were proportional, indicating that a higher dose does not substantially modulate total net bioavailability. Overall, our data shows that brown poplar-type SPE phenolic compounds are highly bioavailable in the form of cinnamic acid and flavonoid conjugates, and that these compounds are rapidly absorbed and eliminated through the urine. Our results suggest that, for a sustained presence in circulation, brown poplar-type propolis supplements should be consumed more than once a day.

Humans

Synergistic effect of flavones and flavonols against herpes simplex virus type 1 in cell culture. Comparison with the antiviral activity of propolis.

The in vitro activity against herpes simplex virus type 1 of the major flavonoids identified in propolis was investigated. Flavonols were found to be more active than flavones, the order of importance being galangin, kaempferol, and quercetin. The efficacy against HSV-1 of binary flavone-flavonol combinations has been also investigated. The synergy demonstrated by all combinations could explain why propolis is more active than its individual compounds.

Animals