Abstract  Cell transformation models have been established for studying the cellular and molecular basis of the neoplastic process. Transformation models have also been utilized extensively for studying mechanisms of chemical carcinogenesis and, to a lesser degree, screening chemicals for their carcinogenic potential. Complexities associated with the conduct of cell transformation assays have been a significant factor in discouraging broad use of this approach despite their reported good predictivity for carcinogenicity. We previously reported that many of the experimental difficulties with the Syrian hamster embryo (SHE) cell transformation assay could be reduced or eliminated by culturing these cells at pH 6.7 culture conditions compared to the historically used pH 7.1-7.3. We and others have shown that morphological transformation (MT), the earliest recognizable phenotype in the multi-step transformation process and the endpoint used in the standard assay to indicate a chemica

Abstract  A polyaniline–ionic liquid (i.e. 1-butyl-3-methylimidazolium tetrafluoroborate, [C b4 smim][BF b4 s]) composite film coated platinum wire (PANI–IL/Pt) was prepared by electrochemical method for the first time. Scanning electron microscopy image showed that the PANI–IL composite film was even and porous. When the PANI–IL/Pt was used as a fiber for the headspace solid-phase microextraction (HS-SPME) of some benzene derivatives (i.e. 1,3-dimethylbenzene, 1,2-dimethylbenzene, 1,4-dichlorobenzene, 1,2-dichlorobenzene, 1,3,5-trimethylbenzene and 1,2,4-trimethylbenzene), followed by gas chromatographic analysis, it presented excellent performance, which was much better than that of PANI/Pt and commercial polydimethylsiloxane fiber. Hence the fiber was coupled with gas chromatography for the determination of these benzene derivatives. It was found that under the optimized conditions the linear ranges were 0.04–400 (So (BgL p-1 s with correlation coefficients above 0.99, the detection limits were 9.3–48.1ngL p-1 s (S/N=3), the relative standard deviations (RSDs) were smaller than 5.1% for five successive measurements with single fiber, and the RSDs for fiber-to-fiber were 5.0–11.1% (n=3) for different benzene derivatives. The proposed method was successfully applied to the extraction and determination of benzene derivatives in waste water and tap water, and the recoveries were 87.1–108.1% for different analytes. Therefore, the PANI–IL/Pt is a promising SPME fiber.

Abstract  The study of the energetics of phenolic compounds has a considerable practical interest since this family of compounds includes numerous synthetic and naturally occurring antioxidants. In this work, density functional theory (DFT) has been used to investigate gas-phase thermochemical properties of the following tri-substituted phenols: 2,4,6-trimethylphenol, 2,6-dimethyl-4-tert-butylphenol, 2, 6-dimethyl-4-methoxyphenol, 2,4,6-tri-tert-butylphenol, 2,6-di-tert-butyl-4-methylphenol, 2,6-di-tertbutyl-4-methoxyphenol, 2,4,6-trimethoxyphenol, 2,6-dimethoxy-4-methylphenol and 2,6-dimethoxy-4-tert-butylphenol. Molecular structures were computed with the B3LYP and the omega B97X-D functionals and the 6-31G(d) basis set. More accurate energies were obtained from single-point energy calculations with both functionals and the 6-311++G(2df,2pd) basis set. Standard enthalpies of formation of the phenolic molecules and phenoxyl radicals were derived using an appropriate homodesmotic reaction. The O-H homolytic bond dissociation enthalpies, gas-phase acidities and adiabatic ionization enthalpies were also calculated. The general good agreement found between the calculated and the few existent experimental gas-phase thermochemical parameters gives confidence to the estimates concerning the phenolic compounds which were not yet experimentally studied. (C) 2014 Elsevier Ltd. All rights reserved.

Abstract  We resolved the controversial assignments of the visible vibronic bands observed from corona discharge of 1,2,3-trimethylbenzene. The vibronic bands belonging to the jet-cooled 2,6-dimethylbenzyl radical were clearly identified from the spectrum observed from corona discharge of 2,6-dimethylbenzyl chloride. After subtracting the bands of the 2,6-isomer from the spectrum observed from corona discharge of 1,2,3-trimethylbenzene, the vibronic bands of the 2,3-isomer were also identified. By comparing data with the known vibrational data of 1,2,3-trimethylbenzene and the results of ab initio calculations, we determined the electronic energies of the D-1 -> D-0 transitions and vibrational mode frequencies in the ground electronic states of the 2,3-and 2,6-dimethylbenzyl radicals.

Abstract  Secondary organic aerosol (SOA) from the photooxidation of aromatic compounds is a very complex mixture containing products with a different chemical nature that are dependent on aging processes. Aging of SOA particles formed from OH-initiated oxidation of benzene was investigated in a home-made smog chamber in this study. The chemical composition of aged benzene SOA particles were measured using aerosol laser time-of-flight mass spectrometer (ALTOFMS) coupled with Fuzzy C-Means (FCM) clustering algorithm. Experimental results showed that nitrophenol, dinitrophenol, nitrocatechol, dinitrocatechol, 6-oxo-2,4-hexadienoic acid, 2,4-hexadiendioic acid, 2,3-dihydroxy-6-oxo-4-hexenoic acid, 2,3-epoxy-4-hexendioic acid, 2,3-epoxy-4,5-dihydroxy-hexanedioic acid and high-molecular-weight (HMW) components were the predominant products in the aged particles. Compared to offline method such as liquid chromatography mass spectrometry (LC-MS) measurement, the real-time ALTOFMS detection approach coupled with the FCM data processing algorithm can make cluster analysis of SOA successfully and provide more information of products. The present results also indicate that benzene SOA aging proceeds through the oxidation of the internal double bond of ring-opened products, phenolic compounds, and acid-catalyzed heterogeneous reactions of carbonyl products. The possible reaction mechanisms leading to these aged products were also discussed and proposed.

Abstract  A series of mesoporous sieves with different pore diameters and structures have been controllably synthesized by adding variable amounts of swelling agent 1,3,5-trimethylbenzene in the preparation process for SBA-15. NiMo catalysts supported by mesoporous sieves are prepared by using the newly proposed Mo-based organic-inorganic hybrid nanocrystals as precursors to load Mo and subsequently using wetness impregnation to load Ni. The hydrodesulfurization (HDS) activity of these obtained catalysts is evaluated in a continuously flowing tubular fixed-bed microreactor using 1 wt% dibenzothiophene (DM.) in heptane as a model compound. The results show that HDS ratio has gradually increased as the pore diameter of supports is enlarged from 7 to 11 nm with the mesostructure keeping an ordered two-dimensional hexagonal symmetcy. When the structure is in the intermediate stage of structural transition from ordered hexagonal mesostructure to mesocecullar structure, HDS ratio is decreased dramatically due to the disordered and collapsed structure to block the pore channels. After the mesocecullar structure with three-dimensional open pore channels is formed which is more convenient for the molecular diffusion, HDS ratio has increased again as a result It is concluded that both the pore size and window size connecting the adjacent pores play an important role in the HDS activity but the window size is of more significant importance, which is the key point for the enhanced catalytic performance. (c) 2014 Elsevier Ltd. All rights reserved,

Abstract  Lethality changes were investigated during development in 4 h metal exposed Caenorhabditis elegans. Exposure to examined metals caused severe lethality toxicities in L1- and L2-larvae, in L3-larvae exposed to examined metals at concentrations of 50 and 100 mu M and to Pb, Hg, and Cr at the concentration of 2.5 mu M, in L4-larvae exposed to examined metals at concentrations of 50 and 100 mu M, and in adults exposed to Pb, Hg, and Cr at the concentration of 100 mu M. Moreover, the lethality toxicities induced by Pb and Hg in L1 larvae for 4 h could be largely comparable to those in young adults for 24 h.

Abstract  In the present study, we investigated the possibly neurotoxic effects of metal (Pb and Hg) exposure at different developmental stages on neuronal loss in the GABAergic nervous system and synaptic functions in the nematode Caenorhabditis elegans. Our data suggest that neuronal survival in GABAergic neurons and cholinergic transmission were relatively stable during development in nematodes. Moreover, neurodegeneration, as shown by the neuronal loss and dorsal/ventral cord gaps, was more severely induced by Pb and Hg exposure at the L1 through L3 larval stages than at the L4 larval and young-adult stages. Similarly, pre- and postsynaptic functions were more severely impaired by Pb and Hg exposure at the L1 through L3 larval stages than at the L4 larval and young-adult stages. Furthermore, both aldicarb and levamisole resistance were significantly correlated with neuronal loss, dorsal cord gap, and ventral cord gap in Pb- and Hg-exposed nematodes, suggesting that neuronal survival was noticeably correlated with synaptic function in metal-exposed nematodes during development. Therefore, younger (L1-L3) larvae show more sensitivity to neurotoxicity of neuronal survival and synaptic function than L4 larvae and young adult nematodes.

Abstract  The peroxisome proliferator-activated receptor-gamma (PPAR-gamma) is a ligand-dependent nuclear receptor that has been implicated in the modulation of critical aspects of development and homeostasis, including adipocyte differentiation, glucose metabolism and macrophage development and function. PPAR-gamma is activated by a range of synthetic and naturally occurring substances, including antidiabetic thiazolidinediones, polyunsaturated fatty acids, 15-deoxy-delta prostaglandin J2 and components of oxidized low-density lipoprotein, such as 13-hydroxyoctadecadienoic acid (13-HODE) and 15-hydroxyeicosatetraenoic acid (15-HETE). However, the identities of endogenous ligands for PPAR-gamma and their means of production in vivo have not been established. In monocytes and macrophages, 13-HODE and 15-HETE can be generated from linoleic and arachidonic acids, respectively, by a 12/15-lipoxygenase that is upregulated by the TH2-derived cytokine interleukin-4. Here we show that interleukin-4 also induces the expression of PPAR-gamma and provide evidence that the coordinate induction of PPAR-gamma and 12/15-lipoxygenase mediates interleukin-4-dependent transcription of the CD36 gene in macrophages. These findings reveal a physiological role of 12/15-lipoxygenase in the generation of endogenous ligands for PPAR-gamma, and suggest a paradigm for the regulation of nuclear receptor function by cytokines.

Abstract  This paper reviews some illustrative examples of studies of human neurobehavioral effects from experimental as well as long-term occupational organic-solvent exposure. My objective is to present a selection of neurobehavioral solvent studies with the educational aim that we may hopefully learn from the early experiences of others. Some of the methodological problems encountered in these studies are discussed, as well as some reasons for the relative success of the work performed by certain Scandinavian research teams.

Abstract  We assessed the frequencies of micronucleated lymphocytes in 3,036 individuals living in 16 settlements in the western part of the Altai region. The majority of individuals with significantly high frequencies of micronucleated lymphocytes were detected in settlements adjacent to the Semipalatinsk atomic testing ground (SATG). The most considerable genome instability was found in individuals born in the period of intensive testing at the SATG (1949 to 1962). Moreover, we determined that the residents of the settlements adjacent to the SATG have significantly high levels of antibodies to potentially oncogenic Epstein-Barr virus, in addition to high frequencies of micronucleated lymphocytes. The considerable Epstein-Barr virus contamination among the residents in the radiation-polluted zone around the SATG was supposed to be caused by immunodeficiency disorders in these individuals and was correlated with high frequencies of micronucleated cells.

Abstract  BACKGROUND AND AIMS: The consumption of germinated barley foodstuff (GBF) prevents inflammation and diarrhoea in a colitis model. In this study we investigated the mechanism of the preventative effect of GBF on experimental colitis in rats, in view of production of bacterial butyrate and preservation of intestinal barrier function. METHODS: Sprague-Dawley rats administered with diets supplemented with 3.5% dextran sodium sulphate were used as an experimental colitis model. Butyrate was given to rats orally or intracaecally. Intestinal barrier function was estimated by light microscopic observation of the mucosa, intestinal permeability and bacterial translocation. RESULTS: Mucosal damage was reduced by intracaecal administration of butyrate, but not by oral administration. Bacterial butyrate production and reduction of mucosal damage depended on the dose of GBF in diets. The action of endogenous bacterial butyrate, including the reduction of intestinal permeability and bacterial translocation, was inhibited by administration of an inhibitor of beta-oxidation of short-chain fatty acids. CONCLUSIONS: The feeding of GBF promotes bacterial butyrate production and improves intestinal barrier function in rats, resulting in mitigation of experimental colitis.

Abstract  The possible effects of environmental and genetic factors on spontaneous frequencies of sister chromatid exchanges (SCEs) and cells with chromosome aberrations (CAs) in human lymphocytes were investigated by analysing 177 completed families (mother, father and at least one child). After removing the effects of methodological, biological and life-style factors by the use of multifactor analysis of variance (MANOVA), SCEs and CAs residuals were analysed by simple correlation analysis and principal component analysis. SCEs and CAs inter-familiar variability was higher than that found within families. A significant correlation was found between the average SCE frequencies shared by parents (the so-called 'midpoint parents', or 'midparent') and offspring (linear slope b=0.26+/-0.07, p<0.05), but also between mother and father (b=0.23+/-0.11, p<0.05) suggesting the presence of an effective environmental factor. The midparent-offspring correlation was found to be sustained by the mother-offspring relationship (b=0.28+/-0.08, p<0.05), being the father-offspring correlation not significant (b=0.16+/-0.11, p0.05). Concerning CAs, no statistically significant correlation between parents was found, but the strong relationship between mother and offspring was confirmed (b=0.468+/-0.11, p<0.001). The SCEs correlation between mother vs. offspring disappeared for older offspring (over 23 years old). The obtained findings strongly showed that the genetic make-up is barely detectable in the presence of domestic environment factors which are shown to play the major role in determining the interfamilial variability of SCE and CA in a general population. These results strengthen the suitability of the use of SCEs and CAs analysis in human cytogenetic surveillance for the detection of effective environmental factors.

Abstract  This study used 2-D agarose gel techniques to examine the effects of the DNA-strand scission enediyne C-1027 on DNA replication in SV40-infected BSC-1 cells. Replication of SV40 DNA was inhibited by C-1027 to a greater extent than was BSC-1 genomic DNA replication in infected cells. Low nanomolar concentrations (0.2-10 nM) of C-1027 affected a rapid, progressive decrease in SV40 replication activity and replication intermediates (RIs) within 15 min after drug addition. A concurrent decrease in the signal of both the SV40 bubble arc and replication activity with increasing concentrations of C-1027 suggested that C-1027 inhibited initiation of new RIs. Additionally, the reduction in bubble arc signal observed with C-1027 was prevented when elongation of nascent chains was blocked by aphidicolin. Thus, the C-1027-induced disappearance of RIs probably is related to the maturation of preformed replication molecules in the absence of initiation of new RIs. Strand damage to SV40 DNA was barely detectable at concentrations where inhibition of replication activity was nearly complete, indicating that C-1027 replication inhibition occurs in trans.

Abstract  This study is the first detailing drug-induced changes in EBV DNA replication intermediates (RIs). Both EBV replication inhibition and damage induction were studied in latently infected human Raji cells treated with the enediyne DNA strand-scission agent C-1027. Analysis of RIs on two-dimensional agarose gels revealed a rapid loss in the EBV bubble arc. When elongation of nascent chains was blocked by aphidicolin, this loss was inhibited, suggesting that C-1027-induced disappearance of RIs was related to maturation of preformed replication molecules in the absence of initiation of new RIs. C-1027 damage to EBV DNA was limited at concentrations where loss of the bubble arc was nearly complete, and none was detected within the replicating origin (ori P)-containing fragment, indicating that replication inhibition occurred in trans. By contrast, the non-nuclear mitochondrial genome was insensitive to replication inhibition but highly sensitive to damage induced by C-1027. C-1027-induced trans inhibition of nuclear but not mitochondrial DNA replication is consistent with a cell cycle checkpoint response to a DNA-damaging agent. EBV replication and Raji cell growth were inhibited at equivalent C-1027 doses.

Abstract  As a possible lipophilic spin probe of in vivo electron spin resonance (ESR), 3-methoxy carbonyl-2,2,5,5-tetramethyl-pyrrolidine-1-yloxy (MCPROXYL) was examined. The permeability of the blood-brain barrier to this compound was evaluated with a brain uptake index and autoradiography, with result that this probe is well distributed in the brain. The in vivo ESR spectra were measured in the head and the abdomen of MCPROXYL-injected living mice. The rate of signal decay of MCPROXYL in the head measured at one hour after X-irradiation was about 75% of that of the controls. The decrease in the head seems to be related to the early response of the brain to X-irradiation. This is the first report that the behavior of free radical such as MCPROXYL in the brain is influenced by X-irradiation. MCPROXYL is thus useful as a novel spin probe for in vivo ESR to monitor the radiation damage in the brain.

Abstract  The effects of calcium ions and modulators of calcium movement on Bacillus thuringiensis insecticidal protein toxicity were investigated with Sf9 cells (Spodoptera frugiperda, fall armyworm) by a new B. thuringiensis toxicity assay based on measurement of fluorescence of ethidium homodimer, a high-affinity DNA stain. CryIC toxicity was substantially stimulated by extracellular calcium in a dose-dependent way (in the millimolar range), while toxicity enhancement could not be replicated when calcium was replaced by barium. This incremental toxicity was reduced by cobalt and lanthanum ions, two inorganic-calcium transport inhibitors. Methoxyverapamil, a voltage-dependent calcium channel blocker, and nifedipine, an inhibitor of dihydropyridine-sensitive L-type calcium channels, had no effect on CryIC toxin activity, but BAY K 8644, an L-type calcium channel activator, increased CryIC activity at high concentrations of extracellular calcium. While A23187, a calcium ionophore, and TMB-8, an inhibitor of intracellular-calcium mobilization, did not change CryIC-induced mortality, thapsigargin, an inhibitor of calcium uptake in intracellular stores, and more particularly trifluoperazine, which inhibits calcium-calmodulin-dependent processes, increased CryIC-mediated toxicity. The incremental effect of extracellular calcium on CryIC-induced toxicity was consistent with an increased concentration of intracellular calcium.

Abstract  CONTEXT: Use of the quadrivalent meningococcal vaccine for control of outbreaks has increased in recent years, but the efficacy of meningococcal vaccine during mass vaccination campaigns in US civilian populations has not been assessed. OBJECTIVES: To evaluate the efficacy of the quadrivalent meningococcal vaccine against serogroup C meningococcal disease in a community outbreak setting and to evaluate potentially modifiable barriers to vaccination in an area with persistent meningococcal disease following immunization. DESIGN: Matched case-control study of vaccine efficacy using cases of serogroup C meningococcal disease in persons eligible for vaccination during mass vaccination campaigns. Control patients were matched by neighborhood and age. The control group was used to identify possible barriers to vaccination. SETTING: Gregg County, Texas, population 106076, from 1993 to 1995. PARTICIPANTS: A total of 17 case patients with serogroup C meningococcal disease eligible for vaccine and 84 control patients. MAIN OUTCOME MEASURES: Vaccine efficacy and risk factors associated with nonvaccination. RESULTS: Vaccine efficacy among 2- to 29-year-olds was 85% (95% confidence interval, 27%-97%) and did not change in bivariate analyses with other risk factors that were significant in univariate analysis. Among control patients, older age was strongly associated with nonvaccination; vaccination rates for 2- to 4-year-olds, 5- to 18-year-olds, and 19- to 29-year-olds were 67%, 48%, and 20%, respectively (chi2 for linear trend, P=.01). CONCLUSIONS: The meningococcal polysaccharide vaccine was effective against serogroup C meningococcal disease in this community outbreak. Although specific barriers to vaccination were not identified, older age was a risk factor for nonvaccination in the target population of 2- to 29-year-olds. In future outbreaks, emphasis should be placed on achieving high vaccination coverage, with special efforts to vaccinate young adults.

Abstract  OBJECTIVES: This study was carried out to investigate possible immunological changes in workers with prolonged low exposure to inorganic mercury in a fluorescent light bulb factory. METHODS: 29 immunological variables were examined in 34 workers with prolonged low level exposure to inorganic mercury (exposed workers) and 35 unexposed workers as the controls. The selected indicator of mercury exposure was concentration of mercury in the urine (U-Hg), which declined progressively from 36.0 micrograms/l in 1978 to 6.0 micrograms/l in the study year 1994. RESULTS: None of the exposed workers had ever shown signs of either acute or chronic inorganic mercury toxicity or had shown any form of hypersensitivity. The only changes found in the exposed workers, compared with the controls, were a reduction of the cells that express cluster differentiation (CD25,(T activation antigen (Tac antigen))) and concentrations of tumour necrosis factor-alpha (TNF-alpha) in serum. However, the decrease of cells that express CD25 was unrelated to occupational exposure and was, in all likelihood a chance finding. Conversely, the decline in serum TNF-alpha was closely associated with occupational exposure. However, no dose-response relation was found between U-Hg and TNF-alpha concentrations; nor were TNF-alpha concentrations affected by cumulative occupational exposure to inorganic mercury in over 20 years. CONCLUSIONS: Tentatively, we suggest that reduced serum TNF-alpha concentrations might be indicative of an in vivo functional defect of the monocyte macrophage system in this particular group of workers even though they were clinically asymptomatic.

Abstract  A new method to facilitate rapid screening of lignin-degrading microorganisms was developed. Fungal strains are cultivated in tissue culture plates containing C-ring-labeled dehydrogenation polymerizate (DHP) (synthetic lignin). Evolved CO(2) is trapped in barium-saturated filter paper and is detected by exposing the paper to X-ray film. Analysis of the autoradiograms, carried out by density measurement with an image analysis program, allows for a semiquantitative estimation of the amount of CO(2) evolved. The method is especially useful for screening for new, powerful lignin-degrading strains in both man-made and natural environments. It eliminates the need for special equipment for their cultivation and trapping of CO(2) as well as laborious sample analysis. The method has in this study been used to test three novel fungal isolates and a laccaseless mutant of the basidiomycete Pycnoporus cinnabarinus. Their ligninolytic capacities were compared with those of the potent lignin degrader Ceriporiopsis subvermispora.

Abstract  The Fluorescein Leakage (FL) test, a short-term in vitro assay measuring damage on exposure to eye irritants to a transepithelial barrier permeability system, has been assessed as an alternative to the Draize rabbit eye irritation assay in the EU/Home Office international validation trial. The assay is based on the degree of leakage of sodium fluorescein through a confluent layer of Madin-Darby canine kidney epithelial (MDCK) cells grown on a tissue culture insert following exposure to a potential irritant. Several different types of tissue culture inserts with different membrane growth surfaces, pore sizes and pore densities have been employed in the past. Problems with chemical binding and other incompatibilities with insert membranes can affect the sensitivity of MDCK cells. Cationic surfactants in particular appear to interfere with the passage of fluorescein, leading to underestimation of damage to MDCK cellular function. A blind comparison between MDCK cells cultured on Anopore() and the Millicell-HA inserts using the EU/Home Office set of surfactants indicate that the sensitivity of cells cultured on Anopore inserts appeared to correlate more closely with in vivo data. The ability to microscopically evaluate the status of MDCK cultures through Anopore further recommends this membrane as possibly the better choice for the FL assay.

Abstract  There is increasing evidence that vitamin D reduces the risk of many types of cancer. Geographic variations in cancer mortality rates in Spain are apparently linked to variations in solar ultraviolet (UV) irradiances and other factors. Cancer mortality rates for 48 continental Spanish provinces for 1978-1992 were used in linear regression analyses with respect to mortality rates for latitude (an index of solar UVB levels), skin cancer (an index of high cumulative UVB irradiance), melanoma (an index related to solar UV irradiance and several other factors) and lung cancer (an index of cumulative effects of smoking). The 9 cancers with mortality rates significantly correlated with latitude for 1 or both sexes were brain, gastric, melanoma, nonmelanoma skin cancer (NMSC), non-Hodgkin's lymphoma (NHL), pancreatic, pleural, rectal and thyroid cancer. Inverse correlations with latitude were found for laryngeal, lung and uterine corpus cancer. The 17 cancers inversely correlated with NMSC are bladder, brain, breast, colon, esophageal, gallbladder, Hodgkin's lymphoma, lung, melanoma, multiple myeloma, NHL, ovarian, pancreatic, pleural, rectal, thyroid and uterine corpus cancer. The 16 correlated with melanoma are bladder, brain, breast, colon, gallbladder, leukemia, lung, multiple myeloma, NHL, ovarian, pancreatic, pleural, prostate, rectal, renal and uterine corpus cancer. The results for lung cancer were in accordance with the literature. These results provide more support for the UVB/vitamin D/cancer hypothesis and indicate a new way to investigate the role of solar UV irradiance on cancer risk. They also provide more evidence that melanoma and NMSC have different etiologies.