Identification of V-type nerve agents in vapor samples using a field-portable capillary gas chromatography/membrane-interfaced electron ionization quadrupole mass spectrometry instrument with Tri-Bed concentrator and fluoridating conversion tube.

A field-portable gas chromatography-mass spectrometry (GC-MS) system (Hapsite ER) was evaluated for the detection of nonvolatile V-type nerve agents (VX and Russian VX (RVX)) in the vapor phase. The Hapsite ER system consists of a Tri-Bed concentrator gas sampler, a nonpolar low thermal-mass capillary GC column and a hydrophobic membrane-interfaced electron ionization quadrupole mass spectrometer evacuated by a non-evaporative getter pump. The GC-MS system was attached to a VX-G fluoridating conversion tube containing silver nitrate and potassium fluoride. Sample vapors of VX and RVX were converted into O-ethyl methylphosphonofluoridate (EtGB) and O-isobutyl methylphosphonofluoridate (iBuGB), respectively. These fluoridated derivatives were detected within 10 min. No compounds were detected when the VX and RVX samples were analyzed without the conversion tube. A vapor sample of tabun (GA) was analyzed, in which GA and O-ethyl N,N-dimethylphosphoramidofluoridate were detected. The molar recovery percentages of EtGB and iBuGB from VX and RVX vapors varied from 0.3 to 17%, which was attributed to variations in the vaporization efficiency of the glass vapor container. The conversion efficiencies of the VX-G conversion tube for VX and RVX to their phosphonate derivatives were estimated to be 40%. VX and RVX vapors were detected at concentrations as low as 0.3 mg m-3 . Gasoline vapor was found to interfere with the analyses of VX and RVX. In the presence of 160 mg m-3 gasoline, the detection limits of VX and RVX vapor were increased to 20 mg m-3 . Copyright © 2017 John Wiley & Sons, Ltd.

Analysis of the canine IgE-binding epitope on the major allergen (Cry j 1) of Japanese cedar pollen with anti-Cry j 1 monoclonal antibodies.

In our previous study [Immunology 91 (1997) 161] using monoclonal antibodies (mAbs) specific to Cry j 1, a major allergen in Japanese cedar (Cryptomeria japonica) pollen, we identified five independent epitopes (EP-1-EP-5) on the molecule and found that EP-1 and EP-5 are the predominant allergic epitopes for humans and monkeys, respectively. In this study, we analyzed the epitopes recognized by IgE in the sera of 10 dogs sensitive to C. japonica pollen allergen using an IgE-ELISA inhibition method with these mAbs. The IgE reaction patterns varied among dogs. In eight of the 10 dogs, IgE recognized EP-5 which is a predominant allergic epitope for monkeys with the pollenosis. In four dogs, IgE recognized EP-1 which is a predominant allergic epitope for human patients with the pollenosis. In three dogs, IgE recognized EP-4 which is a heat-stable epitope. EP-5 is a predominant allergic epitope for dogs and some, but not all, dogs have IgE reaction patterns to the epitopes similar to those of humans.

A novel cell adhesion inhibitor, K-7174, reduces the endothelial VCAM-1 induction by inflammatory cytokines, acting through the regulation of GATA.

A novel inhibitor for the adhesion of monocytes to cytokine-stimulated endothelial cells, K-7174, was selected by an assay system using the cultured human monocytic cells and human endothelial cells. K-7174 inhibited the expression of vascular cell adhesion molecule-1 (VCAM-1) induced by either tumor necrosis factor alpha or interleukin-1beta, without affecting the induction of intercellular adhesion molecule-1 or E-selectin. K-7174 had no effect on the stability of VCAM-1 mRNA. Electrophoretic mobility shift assay revealed that its inhibitory effect on VCAM-1 induction was mediated by an effect on the binding to the GATA motifs in the VCAM-1 gene promoter region. K-7174 did not influence the binding to any of the following binding motifs: octamer binding protein, AP-1, SP-1, ets, NFkappaB, or interferon regulatory factor. These results suggest that the regulation of GATA binding may become a new target for anti-inflammatory drug development, acting through a mechanism independent from NFkappaB activity.

Determinations of free and bound ethanol, acetaldehyde, and acetate in human blood and urine by headspace gas chromatography.

The improved PCA method leads to accurate measurement of ethanol, acetaldehyde, and acetate in blood and urine by headspace gas chromatography. It is important to prevent the formation of artifactual acetaldehyde from coexistent ethanol. The column used for detection of alcohol metabolites was the fused silica glass capillary column bonded with PEG-20M or the fused silica glass capillary column of Pora PLOT Q. In bound alcohol metabolites, recent measurements of hemoglobin-associated acetaldehyde in blood, and ethanol conjugate and acetaldehyde conjugate in urine are reviewed and described as a marker of alcohol abuse.

Epitope specificity of IgE antibodies to a major allergen (Cry j 1) of Japanese cedar pollen in sera of humans and monkeys with pollinosis.

Japanese cedar (Cryptomeria japonica) pollinosis has been reported to occur naturally in Japanese monkeys (Macaca fuscata) as well as humans. Using monoclonal antibodies (mAb) specific to Cry j 1, a major allergen in Japanese cedar pollen, we identified five independent epitopes (EP-1 to EP-5) on the molecule. The epitopes recognized by IgE antibodies in the sera of humans and monkeys with the pollinosis were analysed by an IgE enzyme-linked immunosorbent assay inhibition method with these mAb. In human patients, the mAb to EP-1 strongly blocked the binding of IgE antibodies in all patients' sera to Cry j 1. The reaction patterns of IgE antibodies in monkeys, however, varied among the troops of monkeys. In some troops, the mAb to EP-1 showed a blocking pattern similar to that for human patients. In other troops, mAb to EP-4 and EP-5 blocked binding of IgE. These results indicate that some, but not all, monkeys have antibody responses to the major allergen similar to those of humans.

Effects of bezafibrate on ethanol oxidation in rats.

Bezafibrate is used to lower serum lipid levels in humans. Fibrate derivatives induce an enzyme participating in the beta-oxidation by peroxisomes. We gave ethanol (2 g/kg) orally to bezafibrate-treated (300 mg/kg) male rats of the Wistar strain. Blood ethanol levels were remarkably lower and ethanol elimination stood at 432.6 mg/kg/hr (control, 336.6 mg/kg/hr) in the bezafibrate group (p < 0.01). Blood acetate levels were conversely higher in the bezafibrate group. The fatty acid beta-oxidation activity of liver peroxisome in bezafibrate-treated, clofibrate-treated, or gamma-linolenic acid-treated rats for 4 days was assayed. The activity was 5.8-fold higher in rats given bezafibrate, 5.4-fold in the clofibrate (p < 0.01), and 2.0-fold in the gamma-linolenic acid (p < 0.05). Alcohol dehydrogenase and aldehyde dehydrogenase activity of cytosol in the liver was not induced by the hypolipidemic drugs, but aldehyde dehydrogenase activity in the liver homogenate was induced. From foregoing results, bezafibrate induced in the organism beta-oxidation by peroxisomes and increased H2O2 production, which led to augmented ethanol metabolism by catalase.

Effects of tea (Camellia sinensis) chemical compounds on ethanol metabolism in ICR mice.

The effects of a green tea (Camellia sinensis) extract on ethanol metabolism in ICR male mice were studied. A crude green tea extract (GTE) and the tea components as (-)-epigallocatechin gallate (EGCg), (-)-epigallocatechin (EGC), and caffeine were administered before the tests. One hour later, the mice were orally given 2g/kg body weight (b.w.) of ethanol (20% ethanol w/v). The results show that the levels in the blood and liver of ethanol and acetaldehyde were lower, and that the levels of acetate and acetone were higher than in the controls orally given 500 mg/kg b.w. of GTE. After the administration of 75 mg/kg b.w. and 225 mg/kg b.w. of EGCg, the acetate and acetone concentrations in the blood and liver were lower than in the controls. The mice given caffeine at the same dose as that in GTE showed almost the same effects as the group treated with GTE. This suggests that EGCg and caffeine, the principal components of GTE, both had an effect on ethanol metabolism.

Effects of bezafibrate on erythrocyte membrane phospholipids in alcohol-treated rats.

Male rats of the Wistar strain were divided 4 groups, and give a liquid diet of control feed, bezafibrate (150 mg/kg), ethanol, and ethanol plus bezafibrate for 5 week. The effect of bezafibrate supplementation on rats fed ethanol was examined in terms of the fatty acid composition of the phospholipids in the erythrocyte membrane. In the phospholipids profiles of erythrocyte membranes, PI was significantly decreased. The decrease in PI caused by bezafibrate appeared to substantially affect the membrane and consequently lead to changes in the membrane anchor. In the fatty acid composition of the PC, C20: 4 was significantly decreased in the group receiving alcohol (p < 0.05) but increased in the groups receiving bezafibrate (p < 0.05). In the fatty acid composition of the PE, C16: 0 was significantly increased in the three groups when compared with the control, and C20: 4 was decreased in the alcohol group (p < 0.05). In the fatty acid of SM and PI, C20: 4 was decreased and C18: 0 increased in the alcohol group. In the PS, C14: 0 was increased in the alcohol group, and decreased in the alcohol plus bezafibrate group (p < 0.01). The levels of arachidonic acid in the total fatty acids that constituted the membrane phospholipids were decreased in the rats given ethanol (p < 0.05). However, arachidonic acid in the group of bezafibrate supplementation on rats fed ethanol were elevated in comparison with the alcohol group (p < 0.05). With decreasing arachidonic acid as a marker of alcohol tissue injury following chronic alcohol intake, the effects of bezafibrate supplementation appear to contribute to membrane fluidity by altering the biochemical flexibility of the membrane.

[An attempt of applying the image processing for the automatic estimation of sampled airborne pollen].

Development of techniques to rapidly and easily estimate an airborne pollen quantity is necessitated in order to make out a appropriate prescription for an allergy patient from medical clinical viewpoints, and in order to research a movement of allergen from medical basic viewpoint. The measurement of airborne pollen quantity required a large labour and time, because the amount of pollen grains is visually measured by naked eye. This paper, as a first step to estimate an airborne pollen quantity full-automatically, discuss techniques to measure the quantity of sampled airborne cedar pollen automatically and rapidly using the image processing techniques. As a result, the following facts are cleared. Automatic measurement is possible to some degree without any special image processing. It is important to eliminate a noise on image as blurs on basefilm for high accuracy measurement. The precision has improved fairly with level correction for image. Sharpening filter is the most appropriate process to improve the accuracy of automatic measurement of sampled airborne pollen. This filtering process has a merit that the operativeness is easy.

[An experiment on drinking using breath alcohol monitor (Alcomed 3010) by an electrochemical method].

A drinking experiment was performed to evaluate the efficiency of a breath alcohol monitor, Alcomed 3010. The ethanol concentrations in blood and breath were determined by gas chromatography, and in particular the breath ethanol concentration was determined with the breath alcohol monitor and by gas chromatography. The results obtained by two methods were compared. Based on the blood and breath ethanol concentrations, the following conclusions were drawn reading the breath alcohol monitor. The monitor has practical merit for determination of the breath ethanol level. It is small, usable anywhere, with little error in determination. In measuring principle, tobacco and acetone did not affected levels with the meter, but methanol, n-propanol and n-butanol affected determinations with the alcohol monitor. The breath (AM)/blood (GC) ethanol ratio was 1:2555. Comparison of the values determined with the alcohol monitor and gas chromatography yielded the equation: y = 0.998 x +/- 0.012 (r = 0.994). When determinations were made on the pure ethanol gas by the meter and gas chromatograph, the equation was: y = 0.974 x +/- 0.021 (r = 0.994). It may be said therefore that the alcohol monitor is both practically and functionally excellent.

[A study for the practical use of the mask fitting tester].

The standards require replaceable dust respirators to be designed so that the wearer can easily check facepiece-to-face fitting at any time. The common practice adopted is an air leakage examination between the facepiece and the face in a negative pressure created by sealling-off inhalation area and breathing-in (called "a negative pressure method"). This method offers only subjective testing made by the wearer himself or herself, no objective testing is possible by the third party including supervisors and hygiene staff. Accordingly, we conducted the practical use test of the Mask Fitting Tester (Model MT-02, Roken type) by letting wearers to use the tester at a sanitaryware plant where workers are well instructed for how to wear respirators and also respirators are used in good care and maintenance. In addition, a questionnaire survey was conducted to investigate how wearers perceive the practical use of the Mask Tester. The following are the conclusions of the study and the survey for the practical use of the Mask Tester. 1) At the first fitness test, when examinees wear the respirator as in the usual manner without given particular instructions, 50% of examinees are found unachieved with the leakage rate of the desired value of 5% of less. 2) All the examinees unachieved were instructed by hygiene staff followed by fitting test to check their leakage rate until they pass the desired value. After repeating this three times, there were no examinees found unachieved. 3) 88.2% of these examinees could achieve the desired value only by adjusting headbands and correcting the position of facepiece under instructions.(ABSTRACT TRUNCATED AT 250 WORDS)

Simultaneous measurement of alcohols and hydrogen cyanide in biological specimens using headspace gas chromatography.

We attempted to analyze biological specimens simultaneously for alcohols and hydrogen cyanide. A headspace gas chromatographic method with thick film wide bore column (PEG 20M) for the simultaneous determinations of methanol, ethanol, n-propanol and hydrogen cyanide in blood has been developed. This method was applied for the determinations of methanol, ethanol and hydrogen cyanide in a forensic autopsy case and animal experiments.

Human monoclonal antibody [1-1-2D] against cancer of the uterine cervix.

To obtain a human monoclonal antibody (h-MAb) against cancer of the uterine cervix, lymphocytes from the regional lymph nodes of 14 patients with cervical cancer were fused with a mouse-human heterohybridoma [II]. Of 6,419 hybridomas, 1,295 produced human immunoglobulins (IgG 670, IgM 737). We isolated clone [1-1-2D], which has produced human IgM with stability for more than a year. This antibody reacted with three of five cell lines of cervical cancer but not with normal fibroblasts. Histoimmunostaining showed positive responses to 9/15 specimens of cervical cancer and in 2/7 specimens from cases of cervical dysplasia. Most of the normal human and fetal tissues showed no positive immune response. The positive immune response of [1-1-2D] to the cell membrane observed by the fluorescence antibody method disappeared after periodate treatment and was weakened by trypsin. Neuraminidase did not affect immune reactivity. This antibody showed a positive response by the thin-layer chromatography immunostaining method and precipitated a glycoprotein having a molecular weight of 38 kD. These results suggested that the epitope of the [1-1-2D] antibody is present on a carbohydrate moiety not containing sialic acid that is carried on protein and lipid moieties.

[Protracted (lasting) presence of Japanese cedar pollen allergen (Cry j I) in house dust].

We investigated the relationship between the amounts of Cry j I in house dust and airborne Cryptomeria japonica pollen in the same location. Cry j I was still detected in house dust collected two weeks after airborne C. japonica pollen had disappeared. Disappearance of Cry j I in house dust coincided with the disappearance of symptoms in the C. japonica pollinosis patients who lived in the same area. Airborne Cupressaceae pollen appeared during the latter half of the C. japonica pollen season. Disappearance of Cupressaceae pollen did not coincided with the disappearance of the symptoms in C. japonica pollinosis patients. Therefore, some symptoms of C. japonica pollinosis patients after C. japonica pollen disappeared from the air may be caused by pollen which had attached to clothes and been brought indoors.

Changes in free and bound alcohol metabolites in the urine during ethanol oxidation.

Free and bound ethanol, acetaldehyde, acetate, acetone and methanol in urine during alcohol oxidation were analyzed by means of a head space gas chromatography. Four healthy male volunteers drank beer for 20 min with 16 ml/kg for non-flushers (A, B) and 8 ml/kg for flushers (C, D). In the urine, the highest bound ethanol levels were between 0.5-1.1 mM for the non-flushers (NF) and 0.2-0.3 mM for the flushers (F). The urine free ethanol levels were 23-70 times as high as bound ethanol levels. The maximum free acetaldehyde in urine was 11-13 microM for the NF and 26-55 microM for the F. The urine bound acetaldehyde levels were 4-5 microM for the NF and 7-15 microM for the F. Urine acetaldehyde existed in free forms at 2.4-3.6 times as high concentrations as in bound forms during ethanol oxidation. The urine free acetate ranged between 0.3-2.0 mM. The bound acetate varied between 0.7-1.1 mM. The urine free methanol at 70-110 microM before the intake increased to 104-180 microM. The bound methanol reached to 78-126 microM from 48-97 microM before the intake. Ethanol levels in the urine were ethanol dose-dependent, whereas it was thought that free and bound acetaldehyde or acetate reflected individual metabolic abilities and not the amount of ethanol consumed.

Effects of seed saponins of Thea sinensis L. (Ryokucha saponin) on alcohol absorption and metabolism.

We evaluated the effects of the seed saponins of Thea sinensis L. on alcohol absorption and metabolism in rats and mice. An ethanolic extract from the seeds of T. sinensis was orally administered to the rats 1 hr before or 0.5 hr after administration of ethanol (2 g/kg), and the blood ethanol assayed 0.5, 1, 2, 3, and 4 hr after ethanol administration. The ethanol level decreased after both pre- and post-administration of the extract. The extract was further purified to obtain a saponin fraction which was orally administered to mice 1 hr before ethanol administration. Blood, liver, and stomach were obtained 0, 1, 3, and 6 hr after ethanol administration, and the ethanol, acetaldehyde, acetate, and acetone concentrations in each specimen were measured by head space gas chromatography. The saponin fraction decreased the ethanol levels in the blood and liver but increased that in the stomach five-fold over the control level, suggesting inhibition of alcohol absorption. The ethanol disappearance time from the blood was shortened, suggesting the promotion of alcohol disappearance. The acetate and acetone levels were unaffected. However, the acetaldehyde level decreased in the blood, liver, and stomach. The decreases in the ethanol and acetaldehyde levels in the liver suggested the protective effects of the seed saponins on the liver. The saponins did not directly inhibit hepatic alcohol dehydrogenase activity. The seed saponins of T. sinensis seem to suppress alcohol absorption by slowing gastric emptying and by inhibiting absorption across the cell membranes of the digestive tract.

Confirmation of the airborne occurrence of micron-size airborne pollen antigen carrying particles by immunoblotting.

We confirmed the existence of micron-size airborne particles carrying major pollen antigen of Cryptomeria japonica (Cry j l) and Lolium perenne (Lol p l) by means of an immunoblotting technique and examination by light microscopy. The size of the spots on a nitrocellulose membrane, marking the presence of antigen on the sampler's tape, varied considerably under light microscopic examination. Spots smaller than pollen grains and micron-size spots were observed together with large spots which seem to have been derived from intact pollen grains. The number of micron-size spots counted under a light microscope did not correspond with the number of spots from pollen grain observable with the naked eyes. This suggests other sources of airborne Cry j l particles than intact pollen grains alone.

A new method of counting airborne Japanese cedar (Cryptomeria japonica) pollen allergens by immunoblotting.

We have devised a new method of counting pollen allergen particles, modified from the fluorescent immunoblotting technique of Schumacher et al. Airborne Japanese cedar pollen allergens collected on Burkard's sampling tape were transferred onto a nitrocellulose membrane. The membrane was then treated with antiallergen mouse monoclonal antibody conjugated with alkaline phosphatase. Pollen allergens were detected as purple spots on the nitrocellulose membrane after phosphate substrate staining was performed. Pollen allergen particles were visible under a stereoscopic microscope or to the naked eye and could thus be counted easily. This new counting method takes less time than previous methods and requires no special skill.

[An enzyme-linked immunosorbent assay for the quantitation of the major allergen from Japanese cedar (Cryptomeria japonica) pollen, Cry j I, using monoclonal antibodies].

We have developed an enzyme-linked immunosorbent assay (ELISA) using two anti-Cry j I monoclonal antibodies (KW-S10 and KW-S131) for the quantitation of the major allergen from Japanese cedar (Cryptomeria Japonica) pollen, Cry j I. Polystyrene microplates coated with KW-S131 were incubated with pollen allergen extracts. Cry j I, which bound to the antibody, was detected with alkaline phosphatase-conjugated KW-S10 using chromogenic enzyme substrate. Cry j I could be measured in concentration of between 0.16 and 2.5 ng/ml by this assay. Intra- and inter-assay coefficients of variation were 1.1-3.5% and 0.9-4.6%, respectively. This assay was considered that it was specific for Cry j I of Japanese cedar pollen because it didn't react with allergens of hinoki (Chamaecypairs obtusa) pollen which have antigenicity in common to Japanese cedar pollen. This assay would be useful for the standardization of Japanese cedar pollen allergen extracts.