Characterization of synthetic turf rubber granule infill in Japan: Volatile organic compounds.

There has been extensive studies on the composition of tires and industrial rubber. However, there is insufficient information on volatile organic compounds (VOCs) emitted from rubber granule products used to fill synthetic turf fields. In this study, we applied a passive sampling method for assessing the VOCs emitted from rubber granule products used for filling synthetic turf fields. We also performed a quantitative component analysis using a gas chromatography-mass spectrometer (GC-MS). The component analysis results of 46 rubber granule-based products showed the predominant presence of benzothiazole and methyl isobutyl ketone. The level of benzene, which the International Agency for Research on Cancer classifies as a substance with sufficient evidence for carcinogenicity to humans, was below the lower quantification limit in the products tested in this study. Our study included most of the rubber granule products used for synthetic turf fields in Japan (>95% of the products in the current domestic market of Japan). Therefore, we obtained a comprehensive overview of the VOCs emitted from the rubber granule-based products used in Japan's synthetic turf fields. Estimating the exposure to these airborne VOCs is essential to evaluate the adverse health effects of the VOCs emitted from these rubber granule-based products. Our sampling method and results can help provide key data for such risk assessment studies in the future.

Free formaldehyde in non-medical face masks purchased from the Japanese market since the COVID-19 outbreak.

Since the Coronavirus Disease 2019 (COVID-19) pandemic began, people have been wearing face masks for many hours every day. As these face masks are in contact with the skin, it is important to pay more attention to their quality and safety. This study examined the concentration of free formaldehyde in 90 non-medical face masks and related products (33 nonwoven, 30 woven cloth, 12 polyurethane, and 15 related products) because formaldehyde is a common contact allergen in textile products. For products consisting of mixed materials, each material was sampled, resulting in 103 samples for analysis. Free formaldehyde (34-239 µg/g) was found in three cloth masks, which consisted of cotton and polyester, with antibacterial and antiviral labeling. It was confirmed that the detected formaldehyde originated from the mask-finishing treatment by a hydrochloric acid extraction discrimination test. These masks may elicit contact dermatitis if the consumers have already been sensitized to formaldehyde. However, the risk of contact dermatitis caused by formaldehyde in masks may be considered low since the frequency of formaldehyde detection in masks in Japan is low.

[Ensuring Traceability Using qNMR in the Quantitative Analysis of Formaldehyde and Acetaldehyde].

　Currently, indoor air quality guidelines for formaldehyde and acetaldehyde are set by the Ministry of Health, Labour and Welfare of Japan. Aldehydes are widely used in adhesives and preservatives, and exposure to these compounds via indoor air is a matter of concern. Considering that contact with indoor air is part of daily life, evaluation of indoor air quality is extremely important. 2,4-Dinitrophenylhydrazine (DNPH) derivatization is widely used for quantitative analysis of aldehydes. A certified reference material with traceability to the International System of Units (SI) is required for this method. However, currently, there are no certified reference materials available for aldehyde-DNPH derivatives, which means that the quantified values obtained by this method are not sufficiently reliable. In this study, we determined the actual content and purity of commercially available aldehyde-DNPH derivatives using 1H-quantitative NMR (qNMR), which can be measured with SI-traceability. Although the commercial DNPH derivatives of formaldehyde and acetaldehyde were low concentration solutions, we were able to determine their purities using 1H-qNMR. Furthermore, we were able to separate and quantify the acetaldehyde isomers generated by the derivatization reaction. In conclusion, it is possible to obtain highly accurate results using 1H-qNMR with commercially available reagents that are not certified metrologically.

A chimeric IgE that mimics IgE from patients allergic to acid-hydrolyzed wheat proteins is a novel tool for in vitro allergenicity assessment of functionalized glutens.

BACKGROUND: Acid-hydrolyzed wheat proteins (acid-HWPs) have been shown to provoke severe allergic reactions in Europe and Japan that are distinct from classical wheat allergies. Acid-HWPs were shown to contain neo-epitopes induced by the deamidation of gluten proteins. However, products with variable rates of deamidation can be found. OBJECTIVES: In this work, we studied the effect of the extent of wheat proteins deamidation on its allergenicity. A recombinant chimeric IgE was produced and compared to patients' IgE for its capacity to assess the IgE-mediated triggering potential of acid-HWPs. METHODS: Sera from acid-HWP allergic patients were analyzed via ELISA and a functional basophil assay for their IgE reactivity to wheat proteins with different deamidation levels. A chimeric mouse/human IgE (chIgE-DG1) specific for the main neo-epitope, QPEEPFPE, involved in allergy to acid-HWPs was characterized with respect to its functionality and its reactivity compared to that of patients' IgE. RESULTS: Acid-HWPs with medium (30%) and high (50-60%) deamidation levels displayed a markedly stronger IgE binding and capacity to activate basophils than those of samples with weak (15%) deamidation levels. The monoclonal chIgE-DG1 allowed basophil degranulation in the presence of deamidated wheat proteins. ChIgE-DG1 was found to mimic patients' IgE reactivity and displayed the same ability to rank acid-HWP products in a degranulation assay. CONCLUSION: Increasing the deamidation level of products from 15% to 60% resulted in an approximately 2-fold increase in their antigenicity and a 100-fold increase in their eliciting potential. The chimeric ChIgE-DG1 may be a useful tool to evaluate functionalized glutens for their allergenic potential. By mimicking patient sera reactivity, chIgE-DG1 also provided data on the patients' IgE repertoire and on the functionality of certain repeated epitopes in gluten proteins.

Anaphylactic augmentation by epicutaneous sensitization to acid-hydrolyzed wheat protein in a guinea pig model.

Recent reports suggest that hydrolyzed wheat protein (HWP) variants such as Glupearl® 19S (GP19S) induce immediate-type hypersensitivity via epicutaneous (EC) sensitization. The identification of strong allergens is a key step in product assessment before commercial launch. However, few reports have described the estimation of actual and potential anaphylactic sensitizing capacity. In this study we assessed the strength of both the actual and potential anaphylactic sensitizing capacity by investigating the immediate-type hypersensitivity inducing potential of HWP compared with gluten. We assessed these strengths via the EC route using an EC or intradermal (ID) sensitization method. We quantified the strength of immediate-type hypersensitivity by evaluating the titer of serum antibodies isolated from sensitized subjects using passive cutaneous anaphylaxis (PCA) reactions. We also evaluated the cross-reactivity between GP19S and gluten. GP19S and gluten applied by both the sensitization methods induced obvious IgG1-mediated PCA reactions. GP19S had stronger sensitizing potential than gluten, according to the serum titers and dye spot diameters. The difference in antibody titers between GP19S and gluten was 16-fold for the EC method versus 2-fold for the ID method. GP19S cross-reacted with gluten. Acid hydrolysis of gluten increased anaphylactic sensitizing capacity in the EC method. To our knowledge, our study is the first to quantitatively confirm that HWP and gluten can induce immediate-type hypersensitivity through an intact skin. These findings suggest that acid-HWP imposes a higher risk of EC sensitization than gluten because of the ease with which the former confers a sensitizing effect through the intact skin.

HCELL Expression on Murine MSC Licenses Pancreatotropism and Confers Durable Reversal of Autoimmune Diabetes in NOD Mice.

Type 1 diabetes (T1D) is an immune-mediated disease resulting in destruction of insulin-producing pancreatic beta cells. Mesenchymal stem cells (MSCs) possess potent immunomodulatory properties, garnering increasing attention as cellular therapy for T1D and other immunologic diseases. However, MSCs generally lack homing molecules, hindering their colonization at inflammatory sites following intravenous (IV) administration. Here, we analyzed whether enforced E-selectin ligand expression on murine MSCs could impact their effect in reversing hyperglycemia in nonobese diabetic (NOD) mice. Although murine MSCs natively do not express the E-selectin-binding determinant sialyl Lewis(x) (sLe(x) ), we found that fucosyltransferase-mediated α(1,3)-exofucosylation of murine MSCs resulted in sLe(x) display uniquely on cell surface CD44 thereby creating hematopoietic cell E-/L-selectin ligand (HCELL), the E-selectin-binding glycoform of CD44. Following IV infusion into diabetic NOD mice, allogeneic HCELL(+) MSCs showed threefold greater peri-islet infiltrates compared to buffer-treated (i.e., HCELL(-) ) MSCs, with distribution in proximity to E-selectin-expressing microvessels. Exofucosylation had no effect on MSC immunosuppressive capacity in in vitro assays; however, although engraftment was temporary for both HCELL(+) and HCELL(-) MSCs, administration of HCELL(+) MSCs resulted in durable reversal of hyperglycemia, whereas only transient reversal was observed following administration of HCELL(-) MSCs. Notably, exofucosylation of MSCs generated from CD44(-/-) mice induced prominent membrane expression of sLe(x) , but IV administration of these MSCs into hyperglycemic NOD mice showed no enhanced pancreatotropism or reversal of hyperglycemia. These findings provide evidence that glycan engineering to enforce HCELL expression boosts trafficking of infused MSCs to pancreatic islets of NOD mice and substantially improves their efficacy in reversing autoimmune diabetes. Stem Cells 2013;33:1523-1531.

[Comprehensive analyses of hydrolyzed wheat protein using shotgun proteomics].

Hydrolyzed wheat protein (HWP; hydrolyzed gluten) is used in various types of products worldwide. Several cases of wheat-dependent, exercise-induced anaphylaxis following exposure to HWP (Glupearl 19S) in cosmetics have been reported. Glupearl 19S was produced from the gluten after partial hydrolysis with hydrogen chloride, and its allergenicity is larger than that of gluten (Adachi R., Allergy 2012;67:1392-9.). It is considered that provocation of allergic manifestations is caused by deamidated gluten in food and/or non-food products. Moreover, an increasing number of studies have shown that HWP can induce IgE-mediated hypersensitivity by skin contact and/or food ingestion. However, the essential molecular properties and profiles of HWP are still unknown. In this study, bioinformatic and multivariate analyses using shotgun proteomics have revealed that 27 proteins significantly decreased in Glupearl 19S compared with intact gluten as shown by the ratio of ion signal intensity of tryptic peptides. In contrast, a single protein significantly increased in HWP compared with intact gluten as shown by the ratio of ion signal intensity of tryptic peptides. Furthermore, we have identified six Glupearl 19S-specific peptides using shotgun proteomics, database searches on Mascot Sequence Query, and de novo sequencing. The six peptides were identified as the specific markers of Glupearl 19S.

New chlorogenin hexasaccharide isolated from Agave fourcroydes with cytotoxic and cell cycle inhibitory activities.

A new chlorogenin hexasaccharide (1) was isolated from leaves of Agave fourcroydes (Agavaceae). The structure of the new saponin was elucidated as chlorogenin 3-O-[alpha-L-rhamnopyranosyl-(1-->4)-beta-D-glucopyranosyl-(1-->3)-[beta-D-glucopyranosyl-(1-->3)-beta-D-glucopyranosyl-(1-->2)]-beta-D-glucopyranosyl-(1-->4)-beta-D-galactopyranoside] (1) by spectroscopic analysis and the result of acidic hydrolysis. The new saponin (1) as well as known hexasaccharides (3 and 5) isolated here showed cytotoxicity against HeLa cells, and 1 exhibited a cell cycle inhibitory effect at the G2/M stage at the concentration of 7.5 and 10 microg/mL.

Oral sensitization of W/W(v) mice with ovalbumin and possible involvement of the decrease in gammadelta-T cells.

Mast-cell-deficient WBB6F1-W/W(v) mice (W/W(v)) and congenic wild-type (+/+) mice were sensitized by oral administration of 0.1 or 1.0 mg ovalbumin (OVA) in the form of gavage every day for 9 weeks, and active systemic anaphylaxis (ASA) was induced by intraperitoneal injection of OVA. Production of OVA-specific IgG1 in response to oral sensitization of the W/W(v) mice was very high, and the production of IL-4, IL-5 and IL-10 by splenocytes re-stimulated with OVA in vitro was increased. These findings suggest that Th2-dominant helper T-cell activation had occurred. By contrast, production of OVA-specific IgG1 was low in +/+ mice, and no significant increase in production of Th2-type cytokines by the splenocytes of +/+ mice was observed. Population analysis in Peyer's patches by flow cytometry revealed that the proportion of the CD11c(+) cell in the W/W(v) mice was slightly increased after antigen stimulation. Analysis of the cell surface markers of intraepithelial lymphocytes (IELs) by flow cytometry showed that the proportion of TCRgammadelta-T cells was extremely lower in the W/W(v) mice, especially in the antigen sensitized group. The proportion of TCRgammadelta-T cells in the splenocytes of W/W(v) mice was also lower than in +/+ mice. Taken together, the above findings indicate that W/W(v) mice seems to be a good model not only for studying the induction mechanism of food allergy but for examining the role of TCRgammadelta-T cells in food-induced hypersensitivity.