Pollen-food allergy syndrome (PFAS) associated with birch and alder pollinosis is becoming a problem in Japan, therefore, it is necessary to find apple cultivars in which Mal d 1, a major PFAS allergen component, is expressed at low levels. Relative quantification of Mal d 1 mRNA in 28 Japanese apple cultivars was performed, and several cultivars with low Mal d 1 expression were identified. As it is the protein and not mRNA that is the allergen, it was necessary to ascertain protein levels. To do this, an ELISA test was constructed for the quantification of Mal d 1 protein levels in apples cultivated in the Nagano prefecture. Mal d 1 extraction solutions were compared, and extraction efficiency was shown to be optimal with minimal sample variation when a composition of 1 mol/L sucrose, 2% polyvinylpolypyrrolidone (PVPP), 2 mmol/L EDTA, and 1 mmol/L diethyldithiocarbamic acid was used. Using this approach, Mal d 1 extraction and quantification were performed on 26 apple cultivars from Nagano prefecture. Of the cultivars tested, 24 were significantly lower in Mal d 1 protein than Sun fuji apples. These low Mal d 1 protein expression cultivars are proposed candidates for producing low-allergenic Japanese apples.

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Isoeugenol (IEG) is a natural alkenylbenzene compound which is used as a flavoring additive in foods. However, it has been shown to be a hepatocarcinogen in male B6C3F1 mice. Although there are negative results in several genotoxicity tests, the genotoxicity of IEG in the livers of male mice has not been investigated. To determine whether a genotoxic mechanism is involved in hepatocarcinogenesis, we carried out histopathological analyses, comprehensive DNA adduct analyses, in vivo mutation assays and global gene expression analyses in the livers of male and female B6C3F1 gpt delta mice treated with IEG by gavage at doses of 0, 150, 300 or 600 mg/kg bw/ day for 13 weeks. IEG induced slight hepatocyte hypertrophy along with liver weight gain in male mice treated with 300 mg/kg bw/day IEG and more, but not in similarly treated female mice. Comprehensive DNA adduct analyses by LC-MS/MS showed no specific DNA adduct formation in the liver, and there were no changes in gpt or Spi- mutant frequencies in the livers. A pathway analysis of mRNA expression data as determined with a cDNA microarray suggested activation of pathways associated with peroxisome proliferator　activated receptor (PPAR) α and γ in the livers of male mice. Overall, our data show a lack of genotoxicity in the mechanisms leading to the hepatocarcinogenesis of IEG in mice, and they suggest the involvement of PPARα and γ pathway activation in this process.

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In the case of antibacterial substances (ASs) detected over the uniform limit (0.01 ppm) in processed foods, the maximum residue limits of ASs in ingredients should be verified. However, it would be impossible to obtain ingredients from the same lot as processed in most cases. Thus, the processing factors (PFs: the ratio of the AS concentrations after the process to those before processing) would be important to estimate the concentration of ASs in ingredients. In this study, the PFs of eight ASs in seafood (sea bream) were estimated using artificial residual samples, which were prepared by injection of eight ASs into seafood and standing at 4℃ overnight. The PFs of eight ASs were estimated by the concentration of ASs in the artificial residual samples before and after the processing (boiling and frying). The concentration of ASs with lower octanol/water partition coefficient (logP_ow) (hydrophilic ASs) more rapidly declined during the boiling process than those of ASs with higher logP_ow (lipophilic ASs). On the contrary to that, the concentration of ASs with higher logP_ow rapidly declined during the frying process than those of ASs with lower logP_ow. The applicability of the PFs in the artificial residual sample was examined by comparison of the concentration of oxytetracycline (OTC) residue in the actual residual sample (yellow tail) before and after the processing. The PFs of OTC in the artificial residual sample could successfully correspond to the PFs of OTC in the actual residual samples. The PFs determined by artificial residual samples could be useful for evaluating the ASs concentration in ingredients from those of processed foods.

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Green tea leaves of Camellia sinensis L. contain 2-4% (dry weight: DW) caffeine, whereas Camellia ptilophylla Chang, called cocoa tea, contains about 4% (DW) theobromine instead of caffeine. Methylxanthines such as caffeine, theobromine, and theophylline are generally known to induce a diuresis. Recently, several reports have suggested that epigallocatechin gallate (EGCG) may influence the effects of caffeine. The above-mentioned Camellia contain up to about 10% (DW) of EGCG in addition to caffeine or theobromine. Therefore, in this study, we evaluated whether the diuretic activity in rats induced by oral administration of caffeine and theobromine (5 mg/kg body weight: b.w.) was affected by the coexistence of EGCG (10 mg/kg b.w.).

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The residual oxytetracycline analysis in cinnamon by dispersive solid-phase extraction using high-performance liquid chromatography (HPLC) was developed and validated. Oxytetracycline was extracted using McIlvaine buffer containing ethylenediaminetetraacetic acid (EDTA-2Na). The removal of matrix components derived from cinnamon was carried out using hexane distribution, solid-phase column extraction, and dispersive solid-phase extraction with Z-sep (zirconia-coated silica gel). After the removal of the solvent, the extract was dissolved in a potassium phosphate solution and analyzed by HPLC with a photodiode array detector (PDA). An interfering peak was not observed in the cinnamon extract chromatograms, indicating the high selectivity of this method. Method validation was performed by recovery tests at the maximum residue limit (MRL) concentration (0.1 µg/g) to evaluate the trueness, repeatability (RSD_r), and with-in laboratory reproducibility (RSD_wr). The validation results exhibited sufficient recovery (range, 79-81%) and precision (RSD_r ≤ 5% and RSD_wr ≤ 7%). These values fulfilled the criteria for the validation guidelines for residual pesticide analysis in Japan. The limit of quantification (LOQ) of this method was 0.03 µg/g in Chinese and Ceylon cinnamon. The time required for the preparation of eight test sample solutions was less than four hours. This method can help inspect residual oxytetracycline in cinnamons.

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Gut microbiota plays an important role in maintaining host health through: (i) supplying various nutrients, (ii) regulating energy balance, (iii) modulating the immune response, and (iv) defensing against pathogens. Several studies have shown that pro-biotics and pre-biotics could modulate microbiota and immune response. In addition, the bioactive metabolites released by pro-biotics, namely post-biotics, have been shown to inhibit pathogen growth, maintain microbiota balance, and regulate an immune response. This review summarizes the studies regarding the impact of pro-biotics, pre-biotics, and post-biotics on gut microbiota and immune systems.

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This article is focused on: (i) specific components of the diet associated with inflammation, especially advanced glycation end products (AGEs) that form during thermal processing of food; (ii) experimental models, epidemiological data, and small clinical trials that suggest an important association between dietary AGEs and development of an inflammatory and pro-oxidative states that are conductive to chronic diseases; (iii) delineation of the pathological mechanisms induced by dietary AGEs, both direct (e.g. non receptor-mediated by MGO) and indirect (receptor-mediated)

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