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Background: Silkworm pupa (SWP) food anaphylaxis has been described frequently in Asian countries. However, false‐positive reactions by skin pricks and serum IgE (sIgE) tests to the extract complicate diagnosis, requiring identification of clinically relevant major allergens. Objectives: In this study, we characterized a novel SWP allergen, Bomb m 4, a 30‐kDa lipoprotein, and evaluated its diagnostic sensitivity. Methods: Bomb m 4 was identified by a proteomic analysis. This recombinant (r)Bomb m 4 was overexpressed in Escherichia coli, and the IgE reactivity by ELISA was compared with other reported allergenic proteins: Bomb m 1 (arginine kinase), 27‐kDa glycoprotein, Bomb m 3 (tropomyosin) using the serum samples from 17 SWP allergic patients and 11 asymptomatic sensitized subjects. Results: rBomb m 4‐specific IgE was recognized by all 17 SWP allergic patients. The 27‐kDa glycoprotein and Bomb m 1 sIgE were found in 35.3% and 0%, respectively, in the SWP allergic patients. ELISA sIgE reactivity increased significantly, when 4 M urea was added in serum samples. However, only 16% inhibition of sIgE reactivity to the whole SWP extract was exhibited by rBomb m 4, whereas more than 93% of self‐inhibition of rBomb m 4 sIgE was obtained, possibly due to the low abundance of Bomb m 4 in the extract. Three linear epitopes (81–95, 191–205 and 224–238 residues) of rBomb m 4 were identified. These epitopes are shown to be released by pepsin digestion. Receiver operator characteristic (ROC) analysis showed the highest diagnostic value of Bomb m 4 followed by Bomb m 1, 27‐kDa glycoprotein and Bomb m 3. Conclusion: Bomb m 4 is the major allergen of SWP allergic patients. It has cryptic epitopes which are exposed to IgE antibodies with digestive enzymes. This recombinant Bomb m 4 allergen permits exact diagnosis of SWP allergy. [ABSTRACT FROM AUTHOR]

IgE reactivity between native Que ac 1 and pollen extract (A), between recombinant Que ac 1 and pollen extract (B), and between native Que ac 1 and recombinant Que ac 1 were compared. At least 3 isoallergens (1.0101, 10201, 10301) are present, and isoallergen Que ac 1.0101 was deposited in GenBank under accession No.MN201198, Que ac 1.0201 under MN201199, and Que ac 1.0301 under MN201200. RQue ac 1 was able to inhibit 67.2% of IgE reaction to the whole pollen extract, while 57.6% was inhibited by rBet v 1, 88.7% by nQue ac 1, and 94.7% by the pollen extract at 10 g of inhibitor concentrations, respectively (Figure 2D). [Extracted from the article]

Allergen extracts are commonly utilized for diagnosis and immunotherapy; however, the stability of protease-rich extracts is important for a precise diagnosis and treatment efficacy. The present study determines the optimal conditions for the storage of German cockroach allergen extract. Cockroach extracts were reconstituted in four buffers: normal saline (NS), 50% glycerol in NS, 0.3% phenol in NS, or 0.3% phenol and 50% glycerol in NS. The extracts in different buffers were stored either at room temperature (18–26°C, RT) or refrigerated (2–8°C). Subsequently, the protein concentration and allergen content (Bla g 1 and Bla g 2) in the extracts were examined for the course of one year. Extract potency was estimated by inhibition ELISA. At least 90.5% protein, 94.4% Bla g 1, 65.2% Bla g 2, and 91.4% potency remained after one year when 50% glycerol NS was added to the extract with refrigeration. However, less than 13.7% protein, 17.1% Bla g 1, 0% Bla g 2 and 32.5% potency were maintained after one year when 50% glycerol NS was not added to the extract and was maintained at RT. The addition of 0.3% phenol NS did not show significant effects on extract stability. The addition of 50% glycerol NS and refrigerated storage temperature were found to be important factors for increasing the shelf life of protease-rich cockroach extract. [ABSTRACT FROM AUTHOR]

Abstract: Aim of the study: We investigated the efficacy of Viola mandshurica W. Becker (VM) ethanolic (EtOH) extract in the treatment of bronchial asthma in an ovalbumin (OVA)-induced asthmatic BALB/c mouse model. Materials and methods: Female BALB/c mice were sensitized with intraperitoneal (i.p.) ovalbumin (OVA) on days 0 and 14, and were next given intranasal OVA on days 28–30. Randomized treatment groups of sensitized mice received VM EtOH extract, dexamethasone, or placebo, orally, from days 28 to 30. Results: VM EtOH extract significantly inhibited increases in total immunoglobulin E (IgE) and cytokines IL-4 and IL-13 levels in serum and bronchoalveolar lavage fluid (BALF), and also effectively suppressed airway hyperresponsiveness (AHR), eosinophilia, and mucus hypersecretion, in mice with OVA-induced asthma. Conclusions: The results suggest that VM EtOH extract and allied extracts could be useful herbal medicines for asthma treatment, and that VM may also be a valuable lead material for anti-asthma drug development. [Copyright &y& Elsevier]

The effects of seven heavy metals on hemolymph and tissue ferritin in Galleria mellonella were examined by western blotting. Hemolymph ferritin was clearly upregulated by Hg, while tissue ferritin was dramatically upregulated by Cd, Cu, Mn, Zn and Ni. The level of hemolymph ferritin for Hg and Cd increased approximately 1.5-fold over normal controls, while the level of tissue ferritin for Cd, Cu, Mn, Zn and Ni increased more than twofold, indicating that the regulation of hemolymph and tissue ferritin differ with the different heavy metals. Compared to normal control insects not fed CdCl2, ferritin subunit mRNA from the Malpighian tubules, muscle, the gut and the integument was clearly downregulated, as determined by semi-quantitative reverse transcription–polymerase chain reaction, but ferritin subunit mRNA from the silk gland and the fat body did not show any change. These results suggest that in G. mellonella, tissue ferritin is responsible for Cd stress and is thus important for survival of the moth, and that tissue and hemolymph ferritin have different ferritin-inducible mechanisms in response to dietary CdCl2. [ABSTRACT FROM AUTHOR]

Aim of the study: Lilium lancifolium is commonly used to treat bronchitis, pneumonia, etc. In this study, we investigated the anti-inflammatory effects of methanol extracts of the root of Lilium lancifolium (LL extracts) in LPS-stimulated Raw264.7 cells. Material and methods: Levels of NO, PGE2 and pro-inflammatory cytokines (IL-6 and TNF-α) in the supernatant fraction were determined using sandwich ELISA. Expression of COX-2 and iNOS, phosphorylation of MAPK subgroups (ERK and JNK), and NF-κB activation in extracts were detected via Western blot and immunocytochemistry assays. Results: The LL extract significantly inhibited NO, PGE2, IL-6 and TNF-α production in LPS-stimulated cells, and suppressed iNOS and COX-2 expression. A mechanism-based study showed that phosphorylation of ERK1/2 and JNK and translocation of the NF-κB p65 subunit into nuclei were inhibited by the LL extract. Furthermore, interleukin-4 and interleukin-13 production in Con A-induced splenocytes was suppressed. Conclusion: These results indicate that anti-inflammatory effects of methanol extracts from Lilium lancifolium are due to downregulation of iNOS and COX-2 via suppression of NF-κB activation and nuclear translocation as well as blocking of ERK and JNK signaling in LPS-stimulated Raw264.7 cells. [Copyright &y& Elsevier]