Abstract  Although p38 mitogen-activated protein kinases (MAPK) play a crucial role in the activation of monocyte-derived dendritic cells (MoDC) by contact sensitizers, the upstream signals of p38 MAPK remain undetermined. To examine whether sensitizers induce redox or oxidative stress in dendritic cells (DC), which subsequently stimulate p38 MAPK, we measured the ratio of the oxidized (GSSG) versus reduced (GSH) form of cellular glutathione in MoDC stimulated with five sensitizers including NiCl2 and 2,4-dinitrochlorobenzene (DNCB) and three non-sensitizers including sodium dodecyl sulfate using colorimetric assays. All the sensitizers, but none of the non-sensitizers at sublethal concentration, reduced the GSH/GSSG ratio, which was accompanied by phosphorylation of p38 MAPK. Treatment with the antioxidant, N-acetyl-L-cysteine, which suppressed the reduction of the GSH/GSSG ratio, abrogated both the phosphorylation of p38 MAPK and the augmentation of CD86 expression. A similar response pattern was observed in THP-1 macrophage-monocyte cells. Unexpectedly, however, formaldehyde (HCHO) reduced the GSH/GSSG ratio in MoDC, but not in THP-1. This finding, in conjunction with the observation that DNCB and NiCl2 reduced the GSH/GSSG ratio at different kinetics, indicated that the sensitizers reduced the GSH/GSSG ratio by a different mechanism. These data suggest that the GSH/GSSG imbalance plays a crucial role in triggering DC maturation by sensitizers.

Abstract  The mouse ear swelling test (MEST) was developed as an alternative to guinea pig models for measuring the contact sensitization potential. However, the MEST relies on the quantitative measurement of ear swelling by micrometer as the means of determining the endpoint. The purpose of this study was to investigate the possibility of using cell proliferation in the ear and lymph node by bromodeoxyuridine (BrdU) immunohistochemistry as a reliable marker for MEST. Female Balb/c mice were treated by the topical application of various sensitizers, 2,4-dinitrochlorobenzene (DNCB), toluene diisocyanate (TDI) and α-hexylcinnamaldehyde (HCA) and an irritant, sodium lauryl sulfate (SLS) following the protocol of MEST. The proliferation of cells in the ear and auricular lymph node was analyzed by BrdU incorporations into cells. There were significant increases in the cell proliferations of the ear and auricular lymph node in mice treated with DNCB and TDI compared to the vehicle control. All allergens and the irritant were correctly identified by the MEST using BrdU immunohistochemistry of lymph node responses. The standard MEST assay showed positive results in the case of the strong sensitizers, DNCB and TDI. However, HCA and SLS were not correctly identified in the ear swelling assay. These results suggest that the measurement of cell proliferation in the auricular lymph node using BrdU immunohistochemistry could provide a reliable marker for MEST.

Abstract  Our results indicate that sex hormones play an important role in the regulation of brain and hepatic GST protein during maturity. The conjugating factor GSH does not appear to be under the influence of sex hormones. These observations are of great significance in view of the possibility of continued exposure to neurotoxic chemicals like DDT and Kepone which can cause significant alterations in levels of sex hormones. A reduced GSH activity could lead to a retarded biotransformation of electrophiles and thus to an enhanced toxicity.

Abstract  Characteristic cytokine secretion profiles, consistent with the selective activation of discrete functional subpopulations of T helper (Th) cells, have been demonstrated following repeated topical exposure of mice to chemical contact or respiratory allergens. Draining lymph node cells (LNC) derived from animals treated with the respiratory allergen trimellitic anhydride (TMA; 10%) expressed high levels of the Th2 cytokines interleukins 4 and 10, but little of the Th1 cell product interferon gamma. Under conditions of exposure of equivalent immunogenicity with respect to LNC proliferation, the contact allergen 2,4-dinitrochlorobenzene (DNCB) provoked the converse pattern of cytokine secretion. The purpose of the present investigations was to examine dose-response relationships with respect to cytokine production with a wider range of chemical allergens. In each case, cytokine secretion patterns were compared with those observed with LNC prepared from animals exposed concurrently to TMA or DNCB. Despite some inter-experimental variation in the absolute amounts of cytokines produced, DNCB- and TMA-activated LNC invariably expressed Th1- and Th2-type patterns, respectively. At all concentrations tested, the contact allergens isoeugenol and formaldehyde stimulated a Th1-type cytokine secretion profile, whereas a Th2-type pattern was induced following exposure to the chemical respiratory allergens cyanuric chloride and diphenylmethane diisocyanate. These data demonstrate that divergent cytokine secretion profiles characterize immune responses to different classes of chemical allergen and suggest that it may be possible, in a single integrated assay, to identify and classify chemical allergens as a function of induced cytokine production patterns.

Abstract  The expression of MHC class II (Ia) antigens on mouse keratinocytes was studied following both the induction and elicitation of contact sensitivity, and after primary irritant reactions. IA+ and IE+ keratinocytes were detected, using an indirect immunofluorescence assay on epidermal sheets, only after the induction and elicitation of contact sensitivity with the sensitizers oxazalone, picryl chloride and 2,4-dinitrochlorobenzene but not with formaldehyde. Ia+ keratinocytes were not detected after epicutaneous application of the non-sensitizing irritants croton oil, SDS and anthralin, or following attempted sensitization of nude mice, suggesting that the expression of Ia antigen on keratinocytes during contact sensitivity reactions is T-cell mediated. Because Ia antigen expression on keratinocytes could be detected only several days after induction or elicitation of contact sensitivity, and contact sensitization could also be demonstrated to occur independently of aberrant Ia expression, Ia+ keratinocytes cannot be involved in the initiation of these reactions. However, they might be important in exerting an immunomodulatory influence during the later stages of the responses to certain sensitizers.

Abstract  2 protocols are presented to predict and assess allergic reactions after application of chemicals or cosmetic products, either raw materials or finished products. The first is an open epidermal induction and challenge, the intensity of the reaction being maximized by injecting Freund's complete adjuvant into the foot pad. The second allows detection of weaker allergens and is a quasi-intradermal induction: both the adjuvant and test substance are injected into the foot pad. In both protocols, the challenge consisted of a single topical application in the lumbar region of 10 microliter of test substance and components, allowing study of cross-sensitization, the large surface available permitting 6 different contacts.

Abstract  1-Chloro-2,4-dinitrobenzene (CDNB) was used to conjugate glutathione (GSH) through the catalysis of lens glutathione S-transferase without the untoward oxidative damage to the lens mediated by GSH oxidants. A 2 hr treatment of the rat lens with 1 mM CDNB resulted in a nearly total depletion of lens GSH with neither formation of GSSG nor glutathione-protein mixed disulfides. Rubidium uptake was found to decrease linearly with the loss of GSH; nevertheless, ionic imbalance did not commence until more than 30% cation pump activity was lost. Glycolytic rate dropped following CDNB treatment, due probably to a decline in demand for ATP by the deactivated cation pump. 31P-NMR studies confirmed the irreversible loss of ATP. CDNB depletion of GSH resulted in a two-fold increase in 14CO2 production from [14C]-1-glucose. Whereas oxidative stress resulted in a six-fold increase in glucose utilization through the hexose monophosphate shunt (HMPS), CDNB-treated lenses showed no such stimulation. This indicated that the residual GSH following CDNB treatment was insufficient for the activation of the glutathione peroxidase-reductase-HMPS mechanism and raised the possibility that the increased glucose utilization might be due to mechanisms other than the HMPS. These results indicate an intimate correlation between the GSH content and major metabolic functions in the lens.

Abstract  Porous polymers with different functional groups were tested as sorbents for off-line preconcentration of nitrobenzenes together with their reduction products from water. In addition to 1,4-di(methacryloyloxymethyl)naphthalene-divinylbenzene porous copolymer, previously used for the solid-phase extraction of aromatic amines, three other porous polymers were used. The additional solid-phase minicolumns were packed with methacrylic ester of p,p'-dihydroxydiphenylpropane diglycidyl ether-divinylbenzene, 4,4'-bismaleimidodiphenylmethane-styrene and non-polar styrene-divinylbenzene. In order to study the sorption properties of these polymeric materials, the recoveries and breakthrough volumes of m-phenylenediamine, o-phenylenediamine, aniline, m-nitroaniline, o-nitroaniline, p-nitroaniline, nitrobenzene, m-dinitrobenzene, o-dinitrobenzene and 1,3,5-trinitrobenzene were determined.

Abstract  The rodent olfactory mucosa is characterized by a mosaic of gene expression that is exhibited among various cell types. Olfactory sensitivity in these animals is conveyed through odorant receptor families that are distinctly expressed within various subsets of the olfactory neuron population. Receptor neurons that express a particular class of odorant receptors exhibit bilaterally symmetric zones, which generally define their location within the nasal cavity. Less characterized are zonal expression profiles of proteins among non-neuronal cell types of the olfactory mucosa. In this study, we survey the expression of three glutathione S-transferase (GST) isozymes (alpha, mu, and pi) in the mouse olfactory mucosa and characterize the zonal expression of the mu isozyme. Immunohistochemistry and Western blot analysis of the GST mu isozyme reveal that the lateral olfactory turbinates I, Ib, II, IIb, and III display a greater intensity of expression for GST mu, in comparison to the dorsal and septal regions of the mucosa. GST alpha and pi isozymes do not display any distinct zonal organization in olfactory tissue of the adult mouse. When the general substrate 1-chloro-2-4-dinitrobenzene (CDNB) was used to assess GST activity within the olfactory tissue, the lateral turbinate regions displayed a higher level of activity when compared to dorsal or septal regions. Analysis of GST mu expression in prenatal and early postnatal olfactory tissue also reveals a zonal expression of the isozyme. We relate the significance of these findings to metabolic topography and olfactory chemosensory function. (C) 2003 Elsevier B.V. All rights reserved.

Abstract  The dose-response relationship (challenge phase) of the skin sensitization response was investigated in previously sensitized Hartley guinea pigs. Larger numbers of animals were used per group at the lower doses so that statistically significant observations could be made. Model compounds known to be skin sensitizers were used: a strong sensitizer, dinitrochlorobenzene (DNCB), and a weaker sensitizer, p-phenylenediamine (PPDA). A gradation in response to changing DNCB doses was easily observed by using either the open epicutaneous test (OET) or the Buehler occlusive patch test. The Buehler test was used to study the dose-response relationship of DNCB sensitization. The sensitivity of the OET and Buehler test was judged not adequate to measure the dose response for PPDA, because at high doses a high incidence of responders was not obtained. Therefore, the maximization test was used to evaluate PPDA. Similar, non-linear dose-response curves were obtained with each compound. The higher doses produced a somewhat linear relationship, but at lower doses the curves flattened out and more slowly approached a zero response. Thus, for potent sensitizers, concentrations found in exposure situations might be in the linear portion of the dose-response curve. For weak responders, use concentrations might be in the shallow portion of the curve, where reactions would be underestimated if a linear dose-response curve were assumed.

Abstract  To better guide the development of liquid chromatography/electron capture-atmospheric pressure photoionization-mass spectrometry (LC/EC-APPI-MS) in analysis of low polarity compounds, the ionization mechanism of 19 compounds was studied using dopant assisted negative ion-APPI. Four ionization mechanisms, i.e., EC, dissociative EC, proton transfer, and anion attachment, were identified as being responsible for the ionization of the studied compounds. The mechanisms were found to sometimes compete with each other, resulting in multiple ionization products from the same molecule. However, dissociative EC and proton transfer could also combine to generate the same [M - H](-) ions. Experimental evidence suggests that O(2)(-*), which was directly observed in the APPI source, plays a key role in the formation of [M - H](-) ions by way of proton transfer. Introduction of anions more basic than O(2)(-*), i.e., C(6)H(5)CH(2)(-), into the APPI source, via addition of di-tert-butyl peroxide in the solvent and/or dopant, i.e., toluene, enhanced the deprotonation ability of negative ion-APPI. Although the use of halogenated solvents could hinder efficient EC, dissociative EC, and proton transfer of negative ion-APPI due to their EC ability, the subsequently generated halide anions promoted halide attachment to compounds that otherwise could not be efficiently ionized. With the four available ionization mechanisms, it becomes obvious that negative ion-APPI is capable of ionizing a wider range of compounds than negative ion chemical ionization (NICI), negative ion-atmospheric pressure chemical ionization (negative ion-APCI) or negative ion-electrospray ionization (negative ion-ESI).

Abstract  BACKGROUND: Offspring of asthmatic mothers have increased risk of developing asthma, based on human epidemiologic data and experimental animal models. The objective of this study was to determine whether maternal allergy at non-pulmonary sites can increase asthma risk in offspring.

METHODS: BALB/c female mice received 2 topical applications of vehicle, dinitrochlorobenzene, or toluene diisocyanate before mating with untreated males. Dinitrochlorobenzene is a skin-sensitizer only and known to induce a Th1 response, while toluene diisocyanate is both a skin and respiratory sensitizer that causes a Th2 response. Both cause allergic contact dermatitis. Offspring underwent an intentionally suboptimal protocol of allergen sensitization and aerosol challenge, followed by evaluation of airway hyperresponsiveness, allergic airway inflammation, and cytokine production. Mothers were tested for allergic airway disease, evidence of dermatitis, cellularity of the draining lymph nodes, and systemic cytokine levels. The role of interleukin-4 was also explored using interleukin-4 deficient mice.

RESULTS: Offspring of toluene diisocyanate but not dinitrochlorobenzene-treated mothers developed an asthmatic phenotype following allergen sensitization and challenge, seen as increased Penh values, airway inflammation, bronchoalveolar lavage total cell counts and eosinophilia, and Th2 cytokine imbalance in the lung. Toluene diisocyanate treated interleukin-4 deficient mothers were able to transfer asthma risk to offspring. Mothers in both experimental groups developed allergic contact dermatitis, but not allergic airway disease.

CONCLUSION: Maternal non-respiratory allergy (Th2-skewed dermatitis caused by toluene diisocyanate) can result in the maternal transmission of asthma risk in mice.

Abstract  These studies were conducted to investigate the potential use of a flow cytometric analysis method for the identification and differentiation of chemicals with the capacity to induce irritation, IgE- or T cell-mediated hypersensitivity responses. An initial study investigated the ability of equally sensitizing concentrations (determined by local lymph node assay) of IgE-mediated (Toluene Diisocyanate-TDI) and T cell-mediated (Dinitrofluorobenzene-DNFB) allergens to differentially modulate the IgE+B220+ population in the lymph nodes draining the dermal exposure site. Sodium lauryl sulfate (SLS) was also tested as a nonsensitizing irritant control. Female B6C3F1 mice were dermally exposed once daily for 4 consecutive days, with the optimum time point for analysis determined by examining the IgE+B220+ population 8, 10, and 12 days post-initial chemical exposure. At the peak time point, day 10, the IgE+B220+ population was significantly elevated in TDI (41%), while moderately elevated in DNFB (18%) exposed animals when compared to the vehicle (0.8%), and remained unchanged in SLS (2.2%) exposed animals when compared to the ethanol control (2.5%). Experiments in our laboratory and others have demonstrated that the draining lymph node B220+ population becomes significantly elevated following exposure to allergens (IgE- and T cell-mediated), not irritants, allowing for their differentiation. An existing mouse ear swelling assay was used to identify chemical irritants. Therefore, using the endpoints of percent ear swelling, percent B220+ cells, and percent IgE+B220+ cells, a combined irritancy/phenotypic analysis assay was developed and tested with tetradecane (irritant), toluene diisocyanate, trimellitic anhydride (IgE-mediated allergens), benzalkonium chloride, dinitrofluorobenzene, oxazolone, and dinitrochlorobenzene (T cell-mediated allergens) over a range of concentrations. Based upon the pattern of response observed, a paradigm was developed for continued evaluation: Irritant exposure will result in significant ear swelling without altering the B220+ or IgE+B220+ populations. Exposure to sensitizers (IgE-mediated or T cell-mediated) will increase the B220+ population and the percent ear swelling will remain unchanged or will significantly increase, depending on the irritancy capacity of the chemical. Both the IgE+B220+ and B220+ populations will become elevated at the same test concentration following exposure to IgE-mediated, hypersensitivity inducing allergens. At its peak, the percent of IgE+B220+ cells will be equal to the percent of B220+ cells. The B220+ population will increase at a lower test concentration than the IgE+B220+ population, following exposure to T cell-mediated, hypersensitivity inducing allergens. At its peak, the percent of IgE+B220+ cells will reach less than half that of the percent of B220+ cells. The irritancy/phenotypic analysis method may represent a single murine assay able to identify and differentiate chemicals with the capacity to induce irritation, or IgE-mediated or T cell-mediated responses.

Abstract  PmGSTBI-1 (Proteus mirabilis glutathione S-transferase B1 1) has two tryptophan residues at positions 97 and 164 in each monomer. Structural data for this bacterial enzyme indicated that Trp(97) is positioned in the helix alpha4, whereas Trp(164) is located at the bottom of the helix alpha6 in the xenobiotic-binding site. To elucidate the role of the two tryptophan residues they were replaced by site-directed mutagenesis. Trp(97) and Trp (164) were mutated to either phenylalanine or alanine. A double mutant was also constructed. The effects of the replacement on the activity, structural properties and antibiotic-binding capacity of the enzymes were examined. On the basis of the results obtained, Trp(97) does not seem to be involved in the enzyme active site and structural stabilization. In contrast, different results were achieved for Trp(164) mutants. Conservative substitution of the Trp(164) with phenylalanine enhanced enzyme activity 10-fold, whereas replacement with alanine enhanced enzyme activity 17- fold. Moreover, the catalytic efficiency for both GSH and 1-chloro-2,4-dinitrobenzene substrates improved. In particular, the catalytic efficiency for 1-chloro-2,4-dinitrobenzene improved for both W164F (Trp(164) --> Phe) and W164A by factors of 7- and 22-fold respectively. These results are supported by molecular graphic analysis. In fact, W164A presented a more extensive substrate-binding pocket that could allow the substrates to be better accommodated. Furthermore, both Trp(164) mutants were significantly more thermolabile than wild-type, suggesting that the substitution of this residue affects the overall stability of the enzyme. Taken together, these results indicate that Trp(164) is an important residue of PmGSTB 1-1 in the catalytic process as well as for protein stability.

Abstract  The vasodilating action of organic nitrates, such as nitroglycerin (NTG), is thought to be mediated through metabolic conversion to nitric oxide (NO) in vascular smooth muscle. Although the pertinent enzyme(s) that carries out this crucial step has not been identified, previous studies have shown that the primary enzymatic site is located within cellular membrane fractions. In these studies, we examined the binding of [14C]NTG to microsomal fractions from bovine coronary arteries. Specific binding was linearly related to protein concentration, and binding equilibrium was reversible, reached equilibrium within 1 hr, and remained stable for 4 hr at 25 degrees. Competition experiments with unlabeled NTG demonstrated the presence of two binding sites of differing affinities (high-affinity site: Bmax 24.1 +/- 0.9 pmol/mg protein, Kd 554 +/- 22 pM; low-affinity site: Bmax 79.0 +/- 2.9 pmol/mg protein, Kd 151 +/- 3 microM). Both of the thiol alkylators 1-chloro-2,4-dinitrobenzene and N-ethylmaleimide were found to inhibit [14C]NTG binding, as well as enzymatic generation of NO from NTG, in a concentration-dependent manner. Competition of [14C]NTG was also observed with five other organic nitrate vasodilators, and the degree of competition was linearly related to the in vitro vaso-relaxing potencies of these agents. Parallel experiments also showed that in the absence of thiol cofactor, the enzymatic production of NO from NTG was antagonized competitively by less potent organic nitrates. Intact blood vessel experiments using rat aorta also showed that the presence of isosorbide dinitrate (ISDN), at concentrations that did not induce relaxation alone, caused a slight but significant shift in the relaxation potency of NTG (EC25 9 +/- 2 versus 28 +/- 7 nM, in the presence and absence of 0.3 microM ISDN, respectively; P < 0.05). These results demonstrate the presence of specific binding of organic nitrates to microsomal proteins in vascular smooth muscle, and the observed binding is apparently related to enzymatic conversion to NO and the vasodilating properties of these compounds.

Abstract  The photocatalytic degradation of nitrobenzene and substituted nitrobenzenes under UV exposure was investigated with combustion synthesized nano- TiO2 and commercial TiO2 catalyst, Degussa P-25. The experimental data indicated that the photodegradation kinetics was first order. The photocatalytic degradation rates were considerably higher when catalyzed with combustion synthesized TiO2 compared to that of Degussa P-25. The degradation rate coefficients followed the order: 1-chloro,14-dinitrobenzene similar or equal to 4-nitrophenot > 2-nitrophenol > 1-chloro.4-nitrobenzene > 3-niti-ophenol > 2,4-dinitrophenol > 1 -chloro,2-nitrobenzene > nitrobenzene > 1,3-dinitrobenzene. Plausible mechanisms and reasons for the observation of the above order are discussed. (c) 2005 Elsevier B.V. All rights reserved.

Abstract  Attachment of glutathione (GSH) to cysteine residues in proteins (S-glutathionylation) is a reversible post-translational modification that can profoundly alter protein structure and function. Often serving in a protective role, for example, by temporarily saving protein thiols from irreversible oxidation and inactivation, glutathionylation can be identified and semiquantitatively assessed using anti-GSH antibodies, thought to be specific for recognition of the S-glutathionylation modification. Here, we describe an alternate mechanism of protein glutathionylation in which the sulfur atoms of the GSH and the protein's thiol group are covalently bound via a cross-linking agent, rather than through a disulfide bond. This form of thiol cross-linking has been shown to occur and has been confirmed by mass spectrometry at the solution chemistry level, as well as in experiments documenting the potent antiproliferative activity of the bis-diazeniumdiolate Double JS-K in H1703 cells in vitro and in vivo. The modification is recognized by the anti-GSH antibody as if it were authentic S-glutathionylation, requiring mass spectrometry to distinguish between them.

Abstract  Melanoma patients experience inferior survival after biochemotherapy when their tumors contain numerous cells expressing the inducible isoform of NO synthase (iNOS) and elevated levels of nitrotyrosine, a product derived from NO. Although several lines of evidence suggest that NO promotes tumor growth and increases resistance to chemotherapy, it is unclear how it shapes these outcomes. Here we demonstrate that modulation of NO-mediated S-nitrosation of cellular proteins is strongly associated with the pattern of response to the anticancer agent cisplatin in human melanoma cells in vitro. Cells were shown to express iNOS constitutively, and to generate sustained nanomolar levels of NO intracellularly. Inhibition of NO synthesis or scavenging of NO enhanced cisplatin-induced apoptotic cell death. Additionally, pharmacologic agents disrupting S-nitrosation markedly increased cisplatin toxicity, whereas treatments favoring stabilization of S-nitrosothiols (SNOs) decreased its cytotoxic potency. Activity of the proapoptotic enzyme caspase-3 was higher in cells treated with a combination of cisplatin and chemicals that decreased NO/SNOs, whereas lower activity resulted from cisplatin combined with stabilization of SNOs. Constitutive protein S-nitrosation in cells was detected by analysis with biotin switch and reduction/chemiluminescence techniques. Moreover, intracellular NO concentration increased significantly in cells that survived cisplatin treatment, resulting in augmented S-nitrosation of caspase-3 and prolyl-hydroxylase-2, the enzyme responsible for targeting the prosurvival transcription factor hypoxia-inducible factor-1α for proteasomal degradation. Because activities of these enzymes are inhibited by S-nitrosation, our data thus indicate that modulation of intrinsic intracellular NO levels substantially affects cisplatin toxicity in melanoma cells. The underlying mechanisms may thus represent potential targets for adjuvant strategies to improve the efficacy of chemotherapy.

Abstract  The threat of terrorism and the need for homeland security calls for advanced technologies to detect the concealed explosives safely and efficiently. We demonstrated highly sensitive and selective detection of traces of nitro-aromatic explosive compounds by functionalizing gallium nitride (GaN) nanowires with titanium dioxide (TiO2) nanoclusters to address this issue. The hybrid sensor devices were developed by fabricating two- terminal devices using individual GaN nanowires (NWs) followed by the deposition of TiO2 nanoclusters (NCs) using sputtering technique. The photo-modulated GaN/TiO2 NWNC hybrids showed remarkable selectivity to benzene and related aromatic compounds, with no measureable response for other analytes at room temperature. This paper presents the sensing characteristics of GaN/TiO2 nanowire-nanocluster hybrids towards the different aromatic and nitroaromatic compounds at room temperature. The GaN/TiO2 hybrids were able to detect trinitrotoluene (TNT) concentrations as low as 500 pmol/mol (ppt) in air and dinitrobenzene concentrations as low as 10 nmol/mol (ppb) in air in approximately 30 s. The noted sensitivity range of the devices for TNT was from 8 ppm down to as low as 500 ppt. The detection limit of Dinitrotoluene, nitrobenzene, nitrotoluene, toluene and benzene in air is 100 ppb with a response time of similar to 75 s. The devices show very sensitive and selective response to TNT when compared to interfering compounds like toluene. Integration of this nano-scale technology could lead to tiny, highly sensitive, selective, low-power and smart explosive detectors that could be manufactured cheaply in large numbers.

Abstract  Aims: To examine the existing literature to determine the degree to which percutaneous absorption data obtained using the excised human skin model match those obtained from living man. Methods: The scientific literature was reviewed to collect data on compounds whose percutaneous absorption through human skin had been measured under both in vitro and in vivo conditions. The in vitro-in vivo (IVIV) correlation was evaluated by computing the in vitro/in vivo ratio using total absorption (percent of applied dose) as the metric for comparison. Results: A total of 92 data sets were collected from 30 published studies. The average IVIV ratio across all values was 1.6, though for any single data set there could be a nearly 20-fold difference between the in vitro and in vivo values. In 85% of the cases, however, the difference was less than 3-fold. The correlation was significantly improved when data were excluded from studies in which the protocols for both studies were not fully harmonized. For harmonized data sets the average IVIV ratio was 0.96 and there was a less than 2-fold difference between the in vitro and in vivo results for any one compound, with IVIV ratios ranging from 0.58 to 1.28. The dominant factors leading to exclusion of data were the use of skin from different anatomical sites and vehicles of differing composition. Conclusions: Percutaneous absorption data obtained from the excised human skin model closely approximate those obtained from living man when the two study protocols are appropriately matched. Copyright (C) 2011 S. Karger AG, Basel

Abstract  The hydrogenation of m-dinitrobenzene to m-phenylenediamine in liquid phase was studied with the nickel catalysts supported on SiO2, TiO2, gamma-Al2O3, MgO and diatomite carriers. Based on the experiments of X-ray diffraction (XRD), X-ray photoelectron spectroscopy (XPS), temperature-programmed reduction (TPR), temperature-programmed desorption of hydrogen (H-2-TPD) and activity evaluation, the physico-chemical and catalytic properties of the catalysts were investigated. Among the catalysts tested, the SiO2 supported nickel catalyst showed the highest activity and selectivity towards m-phenylenediamine, over which 97.3% m-dinitrobenzene conversion and 95.1% m-phenylenediamine yield were obtained at 373K under hydrogen pressure of 2.6MPa after reaction for 6 h when using ethanol as solvent. Although TiO2 and diatomite supported nickel catalysts also presented high activity, they had lower selectivity towards m-phenylenediamine. As for gamma-Al2O3 and MgO supported catalysts were almost inactive for the object reaction. It was shown that both the activity and selectivity of the catalysts were strongly depended on the interaction between nickel and the support. The higher activities of Ni/SiO2, Ni/TiO2 and Ni/diatomite could be attributed to the weaker metal-support interaction, on which Ni species presented as crystallized Ni metal particles. On the other hand, there existed strong metal-support interaction in Ni/MgO and Ni/gamma-Al2O3, which causes these catalysts more difficult to be reduced and the availability of Ni active sites decreased, resulting in their low catalytic activity.

Abstract  Early detection of the sensitizing potential of chemicals is an emerging issue for chemical, pharmaceutical and cosmetic industries. In our institute, an in vitro classification model for prediction of chemical-induced skin sensitization based on gene expression signatures in human CD34+ progenitor-derived dendritic cells (DC) has been developed. This primary cell model is able to closely mimic the induction phase of sensitization by Langerhans cells in the skin, but it has drawbacks, such as the availability of cord blood. The aim of this study was to investigate whether human in vitro cultured THP-1 monocytes or macrophages display a similar expression profile for 13 predictive gene markers previously identified in DC and whether they also possess a discriminating capacity towards skin sensitizers and non-sensitizers based on these marker genes. To this end, the cell models were exposed to 5 skin sensitizers (ammonium hexachloroplatinate IV, 1-chloro-2,4-dinitrobenzene, eugenol, para-phenylenediamine, and tetramethylthiuram disulfide) and 5 non-sensitizers (l-glutamic acid, methyl salicylate, sodium dodecyl sulfate, tributyltin chloride, and zinc sulfate) for 6, 10, and 24 h, and mRNA expression of the 13 genes was analyzed using real-time RT-PCR. The transcriptional response of 7 out of 13 genes in THP-1 monocytes was significantly correlated with DC, whereas only 2 out of 13 genes in THP-1 macrophages. After a cross-validation of a discriminant analysis of the gene expression profiles in the THP-1 monocytes, this cell model demonstrated to also have a capacity to distinguish skin sensitizers from non-sensitizers. However, the DC model was superior to the monocyte model for discrimination of (non-)sensitizing chemicals.

Abstract  At present, the assessment of the allergenic potential of chemicals is carried out using animal models. Over the last decade, several in vitro methods mainly using primary dendritic cells have been proposed to identify the potential of chemicals to induce skin sensitization to meet current animal welfare and public opinions. The major limitations of such tests are the donor-to-donor variability, the low levels in the source, and a possible shortage of human sources. The aim of the present investigation was to establish an in vitro test to identify chemical allergens using the human promyelocytic cell line THP-1 in order to avoid some of these difficulties. We investigated whether the chemokine interleukin-8 or CXCL8 (IL-8) production could provide a methodology for the detection of both respiratory and contact allergens. THP-1 cells were exposed to contact allergens (cinnamaldehyde, dinitrochlorobenzene, nickel sulfate, penicillin G, p-phenylenediamine, tetramethylthiuram disulfide), to respiratory allergens (ammonium hexachloroplatinate, diphenylmethane diisocyanate, trimellitic anhydride) and to irritants (salicylic acid, phenol, sodium lauryl sulphate). Following 48 h of incubation, the release of IL-8 was evaluated by sandwich ELISA. IL-8 production was significantly increased after stimulation with all allergens tested, with the exception of trimellitic anhydride, whereas irritants exposure failed to induce IL-8 release. The lack of IL-8 production by trimellitic anhydride can be explained by the rapid hydrolysis of this chemical in water to trimellitic acid, which is not an allergen. In contrast to IL-8 release, CD54 and CD86 expression did not provide a sensitive method failing to correctly identify approximately 30% of the tested compounds. Although CD86 appears to be a more sensitive marker than CD54 when discriminating allergens from irritants neither of these markers provided robust methodology. We also investigated if a common activation pathway in allergen-induced IL-8 production involving p38 mitogen-activated protein kinase could be identified. By Western blot analysis we could indeed demonstrate p38 activation by all chemical allergens tested and, using the selective p38 MAPK inhibitor SB203580, a significant modulation of allergen-induced IL-8 release could be achieved in all cases. Our data suggests that production of IL-8 by naïve THP-1 cells may represent a promising in vitro model for the screening of potential chemical allergens and activation of p38 MAPK represents a common pathway triggered by allergens.

Abstract  Inflammatory skin diseases account for a large proportion of all skin disorders and constitute a major health problem worldwide. Contact dermatitis, atopic dermatitis, and psoriasis represent the most prevalent inflammatory skin disorders and share a common efferent T-lymphocyte mediated response. Oxidative stress and inflammation have recently been linked to cutaneous damage in T-lymphocyte mediated skin diseases, particularly in contact dermatitis. Insights into the pathophysiology responsible for contact dermatitis can be used to better understand the mechanism of other T-lymphocyte mediated inflammatory skin diseases, and may help to develop novel therapeutic approaches. This review focuses on redox sensitive events in the inflammatory scenario of contact dermatitis, which comprise for example, several kinases, transcription factors, cytokines, adhesion molecules, dendritic cell surface markers, the T-lymphocyte receptor, and the cutaneous lymphocyte-associated antigen (CLA). In vitro and animal studies clearly point to a central role of several distinct but interconnected redox-sensitive pathways in the pathogenesis of contact dermatitis. However, clinical evidence that modulation of the skin's redox state can be used therapeutically to modulate the inflammatory response in contact dermatitis is presently not convincing. The rational for this discrepancy seems to be multi-faceted and complex and will be discussed. © 2001 Elsevier Science Inc.