Studies of royal jelly and associated cross-reactive allergens in atopic dermatitis patients.

Royal jelly (RJ), a creamy substance secreted by honeybees, is the exclusive diet for queen bee differentiation and life maintenance. RJ has been used in cosmetics, beverages, medicines, and supplements worldwide. However, allergy is a concerning issue for RJ, especially in atopic dermatitis (AD) and asthma patients. In some cases, allergic reactions are seen after the first intake of RJ, suggesting the existence of allergens cross-reactive with RJ. Information about the cross-reactive allergens is very important for the safe application of RJ; however, study of this cross-reactivity is quite limited. In this study, we attempted to identify allergens cross-reactive with RJ by using serum samples from 30 AD patients who had never been exposed to RJ. In an enzyme-linked immunosorbent assay (ELISA) experiment, RJ-binding IgE antibodies were detected in the serum of 10 out of 30 patients, and their antibody titers ranged from 4- to 2,048-fold dilution ratios. Additionally, 3 AD patients were determined to be positive in a skin-prick test (SPT) with an RJ solution. Significant correlations were observed between the anti-RJ antibody titer and nonspecific IgE and between the anti-RJ antibody titer and the Eczema Area and Severity Index score. We further examined the cross-reactivity between RJ and 14 typical allergens by using an ELISA-inhibition assay and demonstrated that the following 6 allergens showed cross-reactivity with RJ: the European house dust mite (HDM) (Dermatophagoides pteronyssinus), American HDM (Dermatophagoides farinae), snow crab (Chionocetes spp.), edible crab (Cancer pagurus), German cockroach (Blatella germanica), and honeybee venom (Apis mellifera). In conclusion, people with a history of allergic diseases, including AD, asthma, and allergic rhinitis, should be cautioned against consuming RJ products because of the potential for cross-reactive responses to ensure the safe and successful use of RJ supplements.

Insecticidal application of essential oils loaded polymeric nanoparticles to control German cockroach: Design, characterization and lethal/sublethal effects.

Essential oils (EO) from peppermint, palmarosa, geranium, lavender and rosemary were tested against the German cockroach, Blatella germanica L. (Blattaria: Blatellidae). Peppermint and palmarosa oils were the most effective and were included in a polyethylene glycol 6000 matrix to obtain EO loaded polymeric nanoparticles (EOPN). The physicochemical analyses indicated that, at 7 days postformulation, peppermint EOPN had sizes of 380 nm, the loading efficiency (LE) was 72.25% and the polydispersity index (PDI) was >0.4 (polydisperse sample). Palmarosa EOPN had sizes of 191 nm; LE was 89.75% and PDI was <0.25 (monodisperse sample). Peppermint and palmarosa EOPN enhanced the lethal and sublethal effects of the EO on B. germanica. These results suggest that the newly developed nanoinsecticides could be successfully used to control German cockroach.

Insecticidal Activity of Lamiaceae Plant Essential Oils and Their Constituents Against Blattella germanica L. Adult.

The insecticidal activities of 13 Lamiaceae plant oils and their components against adult German cockroaches, Blattella germanica L. (Blattodea: Blattellidae), were evaluated using fumigant and contact bioassay. Among the tested oils, basil, pennyroyal, and spearmint showed the strongest insecticidal activities against adult B. germanica. Insecticidal activity of pennyroyal was 100% against male B. germanica at 1.25 mg concentration in fumigant bioassay. Basil and spearmint revealed 100% and 100% insecticidal activity against male B. germanica at 5 mg concentration, but their activities reduced to 80% and 25% at 2.5 mg concentration, respectively. In contact, toxicity bioassay, basil, pennyroyal, and spearmint oils exhibited 100%, 100%, and 98% mortality against female B. germanica at 1 mg/♀, respectively. Among the constituents identified in basil, pennyroyal, and spearmint oils, insecticidal activity of pulegone was the strongest against male and female B. germanica.

Repellency of zerumbone identified in Cyperus rotundus rhizome and other constituents to Blattella germanica.

The compounds 1,8-cineole and zerumbone (ZER) from the Cyperus rotundus rhizome along with another 11 previously identified rhizome essential oil constituents and α-humulene, which lacks the only carbonyl group present in ZER, as well as binary mixtures of ZER and seven active compounds were tested for repellency to male B. germanica. The results were compared to N,N-diethyl-3-methylbenzamide (deet). In filter-paper choice tests, ZER was the most repellent compound, and α-humulene was ineffective, which indicates that the α,ß-unsaturated carbonyl group of ZER is a prerequisite component for repellency. At 81.5 µg cm-2, enhanced repellency was produced by binary mixtures of ZER and 1,8-cineole, (+)-dihydrocarvone or (R)-(+)-limonene (70:30, 50:50 and 30:70 ratios by weight). These mixtures were very effective against male B. germanica within 24 h and were more repellent than a single compound or deet alone. The optimum ZER content was determined to be more than 50%. In Ebeling choice box tests at 652.4 µg cm-2, these compounds and deet resulted in complete repellency to intact male B. germanica, while they exhibited 35-47% repellency to antennectomized male one. Mixtures formulated from the active constituents of the C. rotundus rhizome could be useful as potential repellents for controlling B. germanica.

Repellent Activity of Apiaceae Plant Essential Oils and their Constituents Against Adult German Cockroaches.

We evaluated the repellent activity of 12 Apiaceae plant essential oils and their components against male and female adult German cockroaches, Blattella germanica L., to find new natural repellents. Of all the plant essential oils tested, ajowan (Trachyspermum ammi Sprague) and dill (Anethum graveolens L.) essential oils showed the most potent repellent activity against male and female adult German cockroaches. Repellent activities of chemicals already identified in active oils were also investigated. Of the compounds identified, carvacrol, thymol, and R-(-)-carvone showed >80% repellent activity against male and female adult German cockroaches at 2.5 µg/cm2. S-(+)-Carvone, (+)-dihydrocarvone, and terpinen-4-ol showed >70% repellent activity against male and female adult German cockroaches at 10 µg/cm2. Our results indicated that Apiaceae plant essential oils and their constituents have good potential as natural repellents against adult German cockroaches.

Molecular basis of the remarkable species selectivity of an insecticidal sodium channel toxin from the African spider Augacephalus ezendami.

The inexorable decline in the armament of registered chemical insecticides has stimulated research into environmentally-friendly alternatives. Insecticidal spider-venom peptides are promising candidates for bioinsecticide development but it is challenging to find peptides that are specific for targeted pests. In the present study, we isolated an insecticidal peptide (Ae1a) from venom of the African spider Augacephalus ezendami (family Theraphosidae). Injection of Ae1a into sheep blowflies (Lucilia cuprina) induced rapid but reversible paralysis. In striking contrast, Ae1a was lethal to closely related fruit flies (Drosophila melanogaster) but induced no adverse effects in the recalcitrant lepidopteran pest Helicoverpa armigera. Electrophysiological experiments revealed that Ae1a potently inhibits the voltage-gated sodium channel BgNaV1 from the German cockroach Blattella germanica by shifting the threshold for channel activation to more depolarized potentials. In contrast, Ae1a failed to significantly affect sodium currents in dorsal unpaired median neurons from the American cockroach Periplaneta americana. We show that Ae1a interacts with the domain II voltage sensor and that sensitivity to the toxin is conferred by natural sequence variations in the S1-S2 loop of domain II. The phyletic specificity of Ae1a provides crucial information for development of sodium channel insecticides that target key insect pests without harming beneficial species.

Insecticidal and acetylcholine esterase inhibition activity of Asteraceae plant essential oils and their constituents against adults of the German cockroach (Blattella germanica).

The fumigant and contact toxicities of 16 Asteraceae plant essential oils and their constituents against adult male and female Blattella germanica were examined. In a fumigant toxicity test, tarragon oil exhibited 100% and 90% fumigant toxicity against adult male German cockroaches at 5 and 2.5 mg/filter paper, respectively. Fumigant toxicities of Artemisia arborescens and santolina oils against adult male German cockroaches were 100% at 20 mg/filter paper, but were reduced to 60% and 22.5% at 10 mg/filter paper, respectively. In contact toxicity tests, tarragon and santolina oils showed potent insecticidal activity against adult male German cockroaches. Components of active oils were analyzed using gas chromatography, gas chromatography-mass spectrometry, or nuclear magnetic resonance spectrometer. Among the identified compounds from active essential oils, estragole demonstrated potent fumigant and contact toxicity against adult German cockroaches. ß-Phellandrene exhibited inhibition of male and female German cockroach acetylcholinesterase activity with IC50 values of 0.30 and 0.28 mg/mL, respectively.

Insecticidal activity of essential oils from eleven Eucalyptus spp. and two hybrids: lethal and sublethal effects of their major components on Blattella germanica.

The aim of the current study was to evaluate the fumigant activity of the essential oils from 11 species of the genus Eucalyptus and two of their hybrids on first instar of Blattella germanica L. The fumigant activity and repellence of the four major monoterpene components of these essential oils also were tested. Fumigant activity was evaluated by exposing nymphs to the vapors emitted by 50 microl of essential oil or monoterpene in a closed container. The lowest knockdown time 50% (KT50) values, expressed in minutes, were elicited by the essential oils of the Eucalyptus grandis X Eucalyptus tereticornis (57.9) hybrid, Eucalyptus sideroxylon A. Cunn (62.0), E. grandis X Eucalyptus camaldulensis (63.8) hybrid, Eucalyptus viminalis Labill (64.1), Eucalyptus dunnii Maiden (64.5), and Eucalyptus grandis (Hill) ex Maiden (68.7). The KT50 values for the remaining essential oils ranged between 74.5 (E. saligna Smith) and 161.4 min (E. tereticornis Smith). The essential oil from the hybrid E. grandis X E. tereticornis was 3.7 times less toxic than dichlorvos (positive control). The KT50 values of monoterpenes were 38.8 for alpha-pinene, 55.3 for 1,8-cineole, 175.6 for p-cymene, and 178.3 for gamma-terpinene. Alpha-pinene was 2.5 times less toxic than dichlorvos. There was a strong positive correlation between the fumigant activity of essential oils and their corresponding 1,8-cineole and alpha-pinene concentration. Repellency was quantified using a video tracking system. Two concentrations of monoterpenes were studied (7 and 70 microg/cm2). All compounds produced a light repellent effect but only when applied at 70 microg/cm2. In all cases, the repellent effect was less than that produced by the broad-spectrum insect repellent N,N-diethyl-3-methylbenzamide (positive control).

[The research of thrombolytic and hemolysis effect from Eupolyphage fibrinolyric protein and its inhibitory effect on S180 ascites tumor of mice].

OBJECTIVE: Eupolyphage fibrinolyric protein (EFP) was isolated and purified from Eupolyphage sineses, and its thrombolytic effect, hemolysis effect and inhibitory effect on S180 ascites tumor were investigated. METHODS: EFP was isolated and purified by ammonium precipitation and DEAE ion exchange chromatography. It's thrombolytic and hemolysis effect were determined. MTT method and Colony-forming method were used to determine the inhibitory effect on S180 ascites tumor. RESULTS: the EFP was proved to have the effect of Thrombolytic and Hemolysis, and both increased dose-dependently, however at a lower concentration, the EFP had no hemocytolysis. The EFP was also proved the effect of inhibitory on cell proliferation and Colony-forming on S180 ascites tumor of Mice. CONCLUSION: EFP has a strong thrombolytic activity and weak hemolytic, and has inhibitory effect on S180 ascites tumor of mice.

Repellence and toxicity of Schinus molle extracts on Blattella germanica.

The biological activities of ethanol and petroleum ether extracts from leaves and fruits of Schinus molle against adults of Blattella germanica were examined by repellence test and topical application. All extracts produced significant repellent effect and mortality.

[Chemical composition and lethal effect of essential oil from Pimenta racemosa (Myrtales: Myrtaceae) on Blatella germanica (Dictyoptera: Blattellidae)]. / Composición química y efecto letal del aceite esencial de Pimenta racemosa (Myrtales: Myrtaceae) sobre Blattella germanica (Dictyoptera: Blattellidae).

Gas chromatography test and Gas chromatography/mass spectrometry test allowed examining the essential oil from Pimenta racemosa leaves (Mill.) J.W. Moore (Myrtales: Myrtaceae). A total of 26 volatile compounds representing 99.5% of the whole composition were identified, being ternipen 4-ol and 1.8 cineol those having the largest portions. Lab bioassays were performed with 5 different oil doses on Blattella germanica. DL50, 15.55 and DL95 = 48.16% were estimated by a probit-log analysis as well as diagnostic dose of 2 microg/insect of 50% oil that is recommended for field surveillance of the use of various formulations in insect control programs.

Molecular, biochemical and histochemical characterization of two acetylcholinesterase cDNAs from the German cockroach Blattella germanica.

Full length cDNAs encoding two acetylcholinesterases (AChEs; Bgace1 and Bgace2) were cloned and characterized from the German cockroach, Blattella germanica. Sequence analyses showed that both genes possess all the typical features of ace, and that Bgace1 is orthologous to the insect ace1 whereas Bgace2 is to the insect ace2. Transcript level of Bgace1 was significantly higher (c. 10 fold) than that of Bgace2 in all 11 tissues examined, suggesting that Bgace1 likely encodes a predominant AChE. Multiple AChE bands were identified by native polyacrylamide gel electrophoresis and isoelectricfocusing from various tissue preparations, among which ganglia produced distinct two major and two minor AChE bands, indicative of the presence of at least two active AChEs. B. germanica AChEs appeared to be mainly localized in the central nervous system as demonstrated by histochemical activity staining, together with quantitative analysis of Bgace transcripts. Fluorescence in situ hybridization of the 1st thoracic ganglion confirmed that Bgace1 is predominantly transcribed and further showed that its transcript is found in almost entire region of inter or motor neurones including the cell bodies and axonal/dendritic branches. Bgace2 transcript is found only in the subset of neurones, particularly in the cell body. In addition, certain neurones were observed to express Bgace1 only.

Contamination affects the performance of insecticidal baits against German cockroaches (Dictyoptera: Blattellidae).

The effects of contamination of insecticidal bait formulations, by using mint oil and silica aerogel, were evaluated in a series of laboratory experiments against the German cockroach, Blattella germanica (L.). Bait consumption at 3 d for uncontaminated baits ranged from 0.01 g for Avert dust to 0.399 g for Combat dry bait with hydramethylnon presented in a station. LT50 values for uncontaminated gel bait deposits ranged from 0.4 d for PreEmpt containing imidacloprid to 4.1 d for Maxforce containing hydramethylnon. As a group, significantly more gel bait was consumed than solid formulations even when both formulations had similar concentrations of the same active ingredient. As a result, gel baits were significantly more toxic than solid formulations. Application of mint oil directly to bait deposits significantly decreased bait consumption and increased overall LT50 values. When bait formulation types were examined individually, there was no difference in consumption or toxicity between contaminated and uncontaminated gel formulations. Contaminated solid baits, however, had significantly greater LT50 values and less consumption than uncontaminated solid baits. Gel formulations probably absorbed the contaminants and removed them from the surface of the bait deposits resulting in normal bait consumption and toxicity. Gel and solid bait deposits, inside plastic exposure stations or alone with no station and placed onto mint oil-contaminated substrates, had significantly lower bait consumption and greater LT50 values than baits placed on uncontaminated substrates. Contamination of a baited area is more likely than contamination of just the bait deposit and therefore a more realistic test of the effects of contamination on bait performance. The importance of contamination to the performance of cockroach baits is discussed.

Sensitization rates for inhalant allergens in Korea; a multi-center study / 천식및알레르기

BACKGROUND AND OBJECTIVE: There are many differences in common inhalant allergens between countries and regions. We need to determine the major inhalant allergens in this country with geographical characteristics. To observe the major inhalant allergens and establish essential ones for the skin prick test in Korea, a nation-wide multicenter study was performed. METHOD: The skin prick tests were performed with the same kits of 43 allergens on 2,554 allergy patients who visited their regional hospitals for one year. RESULT: There are significant differences in the sensitization rate to common inhalant allergens among the centers. Twenty three allergens showed > or=5% sensitization rate: D. pteronyssinus, D. farinae, Tyrophagus putrescentiae, Tetranychus urticae, Panonychus citri, Alternaria alternata, Cladosporium tenuis, cat fur, dog hair, American and German cockroach, tree pollen mixture I, alder, hazel, tree pollen mixture II, birch, beech, oak, plane tree, ragweed, mugwort, hop Japanese and chrysanthemum. CONCLUSION: We suggest the major allergens such as D. pteronyssinus, D. farinae, T. putrescentiae, T. urticae, P. citri, outdoor molds, indoor molds, cat fur, dog hair, German and American cockroaches, tree pollens, mugwort, and hop Japanese pollen should be included in skin test battery in Korea.

Fumigant activity of (E)-anethole identified in Illicium verum fruit against Blattella germanica.

The insecticidal activities of materials derived from the fruit of star anise, Illicium verum, against adults of Blattella germanica were examined by direct contact application and fumigation methods, and compared with those of DDVP, deltamethrin and hydramethylnon. The biologically active constituent of the Illicium fruit was characterized as the phenylpropene, (E)-anethole, by spectroscopic analysis. In a filter paper diffusion test with females, (E)-anethole caused 80.3% mortality at 0.159 mg cm-2 at 1 and 3 days after treatment (DAT), whereas 16.7% mortality at 3 DAT was achieved at 0.079 mg cm-2. DDVP and deltamethrin gave > 90% mortality at 0.019 mg cm-2 at 1 DAT. At 0.009 mg cm-2, DDVP and deltamethrin showed 73.3 and 60% mortality at 1 DAT, respectively, but 93.3 and 76.7% mortality at 3 DAT. Hydramethylnon exhibited 0 and 93.3% mortality at 0.159 mg cm-2 at 1 and 3 DAT, respectively, whereas 6.7% mortality at 3 DAT was observed at 0.079 mg cm-2. In a fumigation test with females, (E)-anethole was much more effective in closed cups than in open ones, indicating that the insecticidal activity of the compound was largely attributable to fumigant action. (E)-Anethole and DDVP caused 100% mortality at 0.398 and 0.051 mg cm-2 4 and 1 h after treatment, respectively. (E)-Anethole showed 46.7% mortality at 0.199 mg cm-2 at 3 DAT, whereas deltamethrin and hydramethylnon at 0.796 mg cm-2 was ineffective for 3-day period. As naturally occurring insect-control agents, the I verum fruit-derived materials described could be useful for managing populations of B germanica.

Sequences of cDNAs encoding calmodulin, and partial structures of calmodulin kinase, and a calcium channel of kdr-resistant and -susceptible German cockroaches, Blattella germanica.

Complementary DNA sequences of genes encoding calmodulin, partial structures of calmodulin-dependent protein kinase II (CaM-kinase II) and an L-type-like calcium channel al subunit (IVS5-IVS6-EF hand region) were identified and compared between susceptible and kdr strains of German cockroach, Blattella germanica. For this purpose, polymerase chain reactions (PCR) were used to obtain their sequences using cDNA from poly(A) + RNA isolated from their heads and thoraces. No mutation differences were found in all three sequences of calcium-regulating proteins between susceptible and strain. Northern blot analysis, however, showed reduced expressions of CaM-kinase II mRNA in two kdr strains. Western blot analysis with an antibody preparation against CaM-kinase II on protein levels confirmed the above strain difference in the titer of this enzyme. In contrast, the levels of calmodulin as well as that of an L-type-like calcium channel gene expression were not different between susceptible and kdr strains.