Mitochondrial translational defect extends lifespan in C. elegans by activating UPRmt.

Aminoacyl-tRNA synthetases (aaRSs) are indispensable players in translation. Usually, two or three genes encode cytoplasmic and mitochondrial threonyl-tRNA synthetases (ThrRSs) in eukaryotes. Here, we reported that Caenorhabditis elegans harbors only one tars-1, generating cytoplasmic and mitochondrial ThrRSs via translational reinitiation. Mitochondrial tars-1 knockdown decreased mitochondrial tRNAThr charging and translation and caused pleotropic phenotypes of delayed development, decreased motor ability and prolonged lifespan, which could be rescued by replenishing mitochondrial tars-1. Mitochondrial tars-1 deficiency leads to compromised mitochondrial functions including the decrease in oxygen consumption rate, complex Ⅰ activity and the activation of the mitochondrial unfolded protein response (UPRmt), which contributes to longevity. Furthermore, deficiency of other eight mitochondrial aaRSs in C. elegans and five in mammal also caused activation of the UPRmt. In summary, we deciphered the mechanism of one tars-1, generating two aaRSs, and elucidated the biochemical features and physiological function of C. elegans tars-1. We further uncovered a conserved connection between mitochondrial translation deficiency and UPRmt.

Cell wall glycosylation in Staphylococcus aureus: targeting the tar glycosyltransferases.

Peptidoglycan (PG) is the major structural polymer of the bacterial cell wall. The PG layer of gram-positive bacterial pathogens such as Staphylococcus aureus (S. aureus) is permeated with anionic glycopolymers known as wall teichoic acids (WTAs) and lipoteichoic acids (LTAs). In S. aureus, the WTA backbone typically consists of repeating ribitol-5-phosphate units, which are modified by enzymes that introduce glycosylation as well as amino acids at different locations. These modifications are key determinants of phage adhesion, bacterial biofilm formation and virulence of S. aureus. In this review, we examine differences in WTA structures in gram-positive bacteria, focusing in particular on three enzymes, TarM, TarS, and TarP that glycosylate the WTA of S. aureus at different locations. Infections with S. aureus pose an increasing threat to human health, particularly through the emergence of multidrug-resistant strains. Recently obtained structural information on TarM, TarS and TarP has helped to better understand the strategies used by S. aureus to establish resistance and to evade host defense mechanisms. Moreover, structures of complexes with poly-RboP and its analogs can serve as a platform for the development of new inhibitors that could form a basis for the development of antibiotic agents.

Experimental methods for the Palaeolithic dry distillation of birch bark: implications for the origin and development of Neandertal adhesive technology.

The destructive distillation of birch bark to produce tar has recently featured in debates about the technological and cognitive abilities of Neandertals and modern humans. The abilities to precisely control fire temperatures and to manipulate adhesive properties are believed to require advanced mental traits. However, the significance given to adhesive technology in these debates has quickly outgrown our understanding of birch bark tar and its manufacture using aceramic techniques. In this paper, we detail three experimental methods of Palaeolithic tar production ranging from simple to complex. We recorded the fuel, time, materials, temperatures, and tar yield for each method and compared them with the tar known from the Palaeolithic. Our results indicate that it is possible to obtain useful amounts of tar by combining materials and technology already in use by Neandertals. A ceramic container is not required, and temperature control need not be as precise as previously thought. However, Neandertals must have been able to recognize certain material properties, such as adhesive tack and viscosity. In this way, they could develop the technology from producing small traces of tar on partially burned bark to techniques capable of manufacturing quantities of tar equal to those found in the Middle Palaeolithic archaeological record.

Production of aromatics through current-enhanced catalytic conversion of bio-oil tar.

Biomass conversion into benzene, toluene and xylenes (BTX) can provide basic feedstocks for the petrochemical industry, which also serve as the most important aromatic platform molecules for development of high-end chemicals. Present work explored a new route for transformation of bio-oil tar into BTX through current-enhanced catalytic conversion (CECC), involving the synergistic effect between the zeolite catalyst and current to promote the deoxygenation and cracking reactions. The proposed transformation shows an excellent BTX aromatics selectivity of 92.9 C-mol% with 25.1 wt.% yield at 400 °C over usual HZSM-5 catalyst. The study of the model compounds revealed that the groups such as methoxy, hydroxyl and methyl in aromatics can be effectively removed in the CECC process. Present transformation potentially provides an important approach for production of the key petrochemicals of BTX and the overall use of bio-oil tar derived from bio-oil or biomass.

Reduced exposure evaluation of an Electrically Heated Cigarette Smoking System. Part 8: Nicotine bridging--estimating smoke constituent exposure by their relationships to both nicotine levels in mainstream cigarette smoke and in smokers.

A modeling approach termed 'nicotine bridging' is presented to estimate exposure to mainstream smoke constituents. The method is based on: (1) determination of harmful and potentially harmful constituents (HPHC) and in vitro toxicity parameter-to-nicotine regressions obtained using multiple machine-smoking protocols, (2) nicotine uptake distributions determined from 24-h excretion of nicotine metabolites in a clinical study, and (3) modeled HPHC uptake distributions using steps 1 and 2. An example of 'nicotine bridging' is provided, using a subset of the data reported in Part 2 of this supplement (Zenzen et al., 2012) for two conventional lit-end cigarettes (CC) and the Electrically Heated Cigarette Smoking System (EHCSS) series-K6 cigarette. The bridging method provides justified extrapolations of HPHC exposure distributions that cannot be obtained for smoke constituents due to the lack of specific biomarkers of exposure to cigarette smoke constituents in clinical evaluations. Using this modeling approach, exposure reduction is evident when the HPHC exposure distribution curves between the MRTP and the CC users are substantially separated with little or no overlap between the distribution curves.

Development of a method for the determination of 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol in urine of nonsmokers and smokers using liquid chromatography/tandem mass spectrometry.

4-(Methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL) is an efficient biomarker of tobacco-specific carcinogen 4-(Methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK). The ability to monitor biomarker concentrations is very important in understanding potential cancer risk. An analytical method using molecularly imprinted polymer (MIP) column coupled with liquid chromatography/tandem mass spectrometry (LC-MS/MS) for the determination of total NNAL in human urine was developed and validated. The combination of MIP column extraction and LC-MS/MS can provide a high sensitive and relatively simple analytical method. The limit of detection (LOD) was 0.30 pg/ml and analysis time was 6min. The method has been applied to urine samples of 36 nonsmokers and 207 smokers. NNAL was found to be significantly higher in the urine of smokers compared with nonsmokers. Compared with smokers with blended cigarettes, Chinese virginia cigarettes smokers had low urinary NNAL levels. There was a direct association between the 24-h mouth-level exposure of carcinogen NNK from cigarette smoking and the concentration of NNAL in the urine of smokers. However, there was not a positive correlation between urinary total NNAL levels in 24 h and tar. Total urinary NNAL is a valuable biomarker for monitoring exposure to carcinogenic NNK in smokers and in nonsmokers. A prediction model of cigarette smoke NNK and urinary average NNAL levels in 24 h was established (y=2.8987x-245.38, r²=0.9952, n=204).

Electron acceptor-dependent identification of key anaerobic toluene degraders at a tar-oil-contaminated aquifer by Pyro-SIP.

Bioavailability of electron acceptors is probably the most limiting factor in the restoration of anoxic, contaminated environments. The oxidation of contaminants such as aromatic hydrocarbons, particularly in aquifers, often depends on the reduction of ferric iron or sulphate. We have previously detected a highly active fringe zone beneath a toluene plume at a tar-oil-contaminated aquifer in Germany, where a specialized community of contaminant degraders codominated by Desulfobulbaceae and Geobacteraceae had established. Although on-site geochemistry links degradation to sulphidogenic processes, dominating catabolic (benzylsuccinate synthase α-subunit, bssA) genes detected in situ appeared to be more related to those of Geobacter spp. Therefore, a stable isotope probing (SIP) incubation of sediment samples with (13)C(7)-toluene and comparative electron acceptor amendment was performed. We introduce pyrosequencing of templates from SIP microcosms as a powerful new strategy in SIP gradient interpretation (Pyro-SIP). Our results reveal the central role of Desulfobulbaceae in sulphidogenic toluene degradation in situ, and affiliate the detected bssA genes to this lineage. This and the absence of (13)C-labelled DNA of Geobacter spp. in SIP gradients preclude their relevance as toluene degraders in situ. In contrast, Betaproteobacteria related to Georgfuchsia spp. became labelled under iron-reducing conditions. Furthermore, secondary toluene degraders belonging to the Peptococcaceae detected in both treatments suggest the possibility of functional redundancy among anaerobic toluene degraders on site.

Metabolites indicate hot spots of biodegradation and biogeochemical gradients in a high-resolution monitoring well.

Anaerobic degradation processes play an important role in contaminated aquifers. To indicate active biodegradation processes signature metabolites can be used. In this study field samples from a high-resolution multilevel well in a tar oil-contaminated, anoxic aquifer were analyzed for metabolites by liquid chromatography-tandem mass spectrometry and time-of-flight mass spectrometry. In addition to already known specific degradation products of toluene, xylenes, and naphthalenes, the seldom reported degradation products benzothiophenemethylsuccinic acid (BTMS), benzofuranmethylsuccinic acid (BFMS), methylnaphthyl-2-methylsuccinic acid (MNMS), and acenaphthene-5-carboxylic acid (AC) could be identified (BFMS, AC) and tentatively identified (BTMS, MNMS). The occurrence of BTMS and BFMS clearly show that the fumarate addition pathway, known for toluene and methylnaphthalene, is also important for the anaerobic degradation of heterocyclic contaminants in aquifers. The molar concentration ratios of metabolites and their related parent compounds differ over a wide range which shows that there is no simple and consistent quantitative relation. However, generally higher ratios were found for the more recalcitrant compounds, which are putatively cometabolically degraded (e.g., 2-carboxybenzothiophene and acenaphthene-5-carboxylic acid), indicating an accumulation of these metabolites. Vertical concentration profiles of benzylsuccinic acid (BS) and methyl-benzylsuccinic acid (MBS) showed distinct peaks at the fringes of the toluene and xylene plume indicating hot spots of biodegradation activity and supporting the plume fringe concept. However, there are some compounds which show a different vertical distribution with the most prominent concentrations where also the precursor compounds peaked.

Tobacco-specific nitrosamine 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL) in smokers in the United States: NHANES 2007-2008.

The tobacco-specific carcinogen 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL), a metabolite of the tobacco-specific nitrosamine (TSNA) 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK), has been measured in urine samples from all participants aged 6 years and older from the National Health and Nutrition Examination Survey 2007-2008. Participants with a serum cotinine concentration of ≥ 10 ng/mL were identified as tobacco users, primarily cigarette smokers. Regression models were developed to calculate geometric mean NNAL concentrations adjusted for serum cotinine, urinary creatinine, cigarettes per day, and Federal Trade Commission tar values of the cigarettes smoked. Significant differences were found by gender (p=0.003) and race/ethnicity (p=0.022 for non-Hispanic white versus non-Hispanic black smokers), but not by menthol type of the cigarettes. Females and non-Hispanic white smokers had the highest adjusted means for urinary NNAL (353 and 336 pg/mL, respectively). The results from this study demonstrated significant relationships between NNAL concentrations and serum cotinine (p<0.001) and urine creatinine (p<0.001). The joint effect of linear and quadratic terms for number of cigarettes smoked per day was also statistically significant (p=0.001). In addition to addressing current NNK exposure levels, these results will form a baseline for future estimates of tobacco users' exposure to this carcinogen.

Sex differences in susceptibility to PAHs is an intrinsic property of human lung adenocarcinoma cells.

Recent epidemiological studies have disputed whether females are at increased risk of lung cancer compared to males. However, several molecular studies are in support of an increased susceptibility to tobacco smoke carcinogens among females. Our earlier findings suggest that women display higher levels of smoking-induced bulky/hydrophobic DNA adducts which may be related to an increased expression of CYP1A1 in their lungs, compared to men. In this in vitro study, 11 lung adenocarcinoma cell lines, 6 of male and 5 of female origin, were exposed to benzo[a]pyrene, cigarette smoke condensate (CSC), or vehicle control. Subsequent expression analysis of genes in the polycyclic aromatic hydrocarbon bioactivation pathway was conducted with Real-Time RT-PCR. DNA adducts were measured in benzo[a]pyrene-exposed cells by ³²P-postlabelling analysis, and CYP1 activity was measured by EROD assay. Analysis of benzo[a]pyrene-DNA adducts showed higher levels of adducts in cell lines from women compared to cell lines from men (p=0.03). The results also revealed significant sex differences in CYP1A1 gene expression, both in untreated cells (p=0.03), and in cells exposed to benzo[a]pyrene (p=0.017) and cigarette smoke condensate (p=0.0043). In CSC-exposed cells, significantly higher levels of CYP1 activity was found in cell lines of female origin (p=0.049). These results are in support of the previously published in vivo data, providing evidence for a higher susceptibility to PAH of women's lungs.

DNA-SIP identifies sulfate-reducing Clostridia as important toluene degraders in tar-oil-contaminated aquifer sediment.

Global groundwater resources are constantly challenged by a multitude of contaminants such as aromatic hydrocarbons. Especially in anaerobic habitats, a large diversity of unrecognized microbial populations may be responsible for their degradation. Still, our present understanding of the respective microbiota and their ecophysiology is almost exclusively based on a small number of cultured organisms, mostly within the Proteobacteria. Here, by DNA-based stable isotope probing (SIP), we directly identified the most active sulfate-reducing toluene degraders in a diverse sedimentary microbial community originating from a tar-oil-contaminated aquifer at a former coal gasification plant. On incubation of fresh sediments with (13)C(7)-toluene, the production of both sulfide and (13)CO(2) was clearly coupled to the (13)C-labeling of DNA of microbes related to Desulfosporosinus spp. within the Peptococcaceae (Clostridia). The screening of labeled DNA fractions also suggested a novel benzylsuccinate synthase alpha-subunit (bssA) sequence type previously only detected in the environment to be tentatively affiliated with these degraders. However, carbon flow from the contaminant into degrader DNA was only ∼50%, pointing toward high ratios of heterotrophic CO(2)-fixation during assimilation of acetyl-CoA originating from the contaminant by these degraders. These findings demonstrate that the importance of non-proteobacterial populations in anaerobic aromatics degradation, as well as their specific ecophysiology in the subsurface may still be largely ungrasped.

Actions of redox-active compound resveratrol under hydrogen peroxide insult in C6 astroglial cells.

The mechanisms by which resveratrol (3,5,4'-trihydroxy-stilbene) imparts neural effects is not well understood. We previously demonstrated that, depending upon the concentration of resveratrol and the cell type, this compound exerts anti-or pro-oxidant effects. In the present study, we investigated the effects of resveratrol on H(2)O(2)-mediated genotoxicity in C6 astroglial cells (I - 1mM H(2)O(2)/30 min or II - 0.1mM H(2)O(2)/6h), evaluated by micronucleus assay, lipid peroxidation (TBARS) and membrane integrity. H(2)O(2) increased micronuclei to 1.5 (I) and 1.7-fold (II), compared to control cells. This DNA damage was prevented (I) or partially prevented (II) by resveratrol. Oxidative insult also increased TBARS, 52% in I and 38% in II, P<0.05. These effects were prevented by resveratrol in I and increased in II (70% of increase). Present data contribute to the understanding of resveratrol effects under oxidative stress damage.

Ebullition-facilitated transport of manufactured gas plant tar from contaminated sediment.

Manufactured gas plant (MGP) tar and wastewater solids historically were discharged into the Penobscot River, Maine,USA, via a sewer at the Bangor Landing site. The tar and wastewater solids accumulated in riverbed sediment over a 5-hectare area downstream from the sewer outfall. Much of the tarry sediment is a hardened mass at the bottom of the river, but in part of the tar deposit (the active zone), the tar remains unhardened. In the active zone, anaerobic biodegradation of organic matter generates methane and carbon dioxide; as gas accumulates and migrates upward, it entrains tar, eventually dragging the tar from the sediment to surface water. Understanding the migration mechanisms in different portions of the tar deposit is critical for modeling the risk posed by the tar at the Bangor Landing site, because during gas-facilitated tar migration, the tar is brought to the water surface, instead of remaining in the sediment. Tar migration from sediment poses a potential human health risk because of the high concentrations of polycyclic aromatic hydrocarbons in the tar. Migration from sediment to the water surface greatly increases the potential exposure of human and ecological receptors to tar that reaches the water surface. In order for tar to migrate from sediment to surface water, three conditions are necessary: the sediment must contain liquid tar, the sediment must produce gas bubbles, and the gas must come into contact with the tarry sediment. Failure to consider facilitated transport of MGP tar from sediment can cause underestimation of site risk and can lead to failure of remedial measures.

The relationship between smoking machine derived tar yields and biomarkers of exposure in adult cigarette smokers in the US.

UNLABELLED: Comprehensive data on human exposure to smoke constituents from different machine-measured tar yield cigarettes is limited. METHODS: This study used a stratified, cross-sectional, multi-center design to estimate biomarkers of exposure (BOE) from nicotine, 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK), pyrene, CO, acrolein, and 1,3-butadiene and their relationship to tar yield categories of cigarette in adult smokers in the U.S. 3625 adults smokers were enrolled into four tar categories < or =2.9 mg (T1), 3.0-6.9 mg (T2), 7.0-12.9 mg (T3), and > or =13.0mg (T4). Biomarkers were measured in blood (carboxyhemoglobin, 4-aminobiphenyl-hemoglobin (4-ABP-Hb)-adducts, serum cotinine) and 24h urine (nicotine and five metabolites, calculated as nicotine equivalents (NE), NNAL, 1-OH-pyrene, 3-HPMA, MHBMA and DHBMA). Data were analyzed using analysis of covariance (ANCOVA). RESULTS: Tar was a significant factor for most biomarkers in the ANCOVA models. The largest least square mean differences between tar categories was 35% for NE per day, 28% for NE per cigarette, 36% for serum cotinine, 42% for NNAL per day, 29% for NNAL per cigarette, 26% for 1-OHP, 24% for COHb, 14% for 3-HPMA and 40% for 4-ABP-Hb. Variability in BOE ranged from 41% to 154% CV. CONCLUSIONS: There was a statistically significant effect of machine-measured tar yield on most BOE, which were generally lower with lower tar yield.

Possible effects on smokers of cigarette mentholation: a review of the evidence relating to key research questions.

Menthol (2-isopropyl-5-methyl-cyclohexan-1-ol) is used in food, pharmaceutical, and tobacco products. Despite its long usage history and GRAS status, scientific literature on effects of cigarette mentholation is limited. Because African-American men have high lung cancer rates and predominantly smoke mentholated cigarettes, and because menthol's cooling effect might affect puffing and smoke inhalation, possible adverse effects of cigarette mentholation have been suggested. We review the evidence on the effects of mentholation on smokers, and we also identify areas for further study. Five large epidemiological studies provide no evidence that cigarette mentholation increases lung cancer risk. Mentholation cannot explain the higher risk for lung cancer in African-American male smokers, who also predominantly smoke mentholated cigarettes. Limited data on other cancers also suggest no risk from mentholation. The scientific literature suggests that cigarette mentholation does not increase puff number or puff volume of smoked cigarettes, and has little or no effect on heart rate, blood pressure, uptake of carbon monoxide, tar intake or retention, or blood cotinine concentration. Mentholation has little effect on other smoke constituents, and no apparent effect on nicotine absorption, airway patency and smoking initiation, dependency or cessation. Any toxicological effects of cigarette mentholation on adult smokers are probably quite limited.

Studies on microbiological degradation of tobacco tar.

A bacterium, strain MR4, was isolated from tobacco tar-contaminated soil and identified as Klebsiella oxytoca based on morphology, physiological and biochemical characteristics and phylogenetic analysis of 16S rDNA sequence. The organism grew optimally at 34 degrees C and pH 7.0 to 7.5. During growth on tobacco tar the isolate produced acid materials, which caused the drop of pH in the cultures. GC/MS analysis indicated that the isolate had the ability to degrade phenolic compounds, heterocyclic compounds, especially nicotine in tobacco tar. The degradation rate of strain MR4 was 75.56% for nicotine, 35.84% to 58.16% for hydroxybenzenes and the other aromatic compounds, 29.15% to 65.56% for heterocyclics, and 35.17% to 82.59% for hydrocarbons.

The loss of retinoic acid receptor alpha, beta and alcohol dehydrogenase3 expression in non-small cell lung cancer.

OBJECTIVE: The reason for unsuccessful retinoid chemoprevention of lung cancer is obscure. Therefore we examined the retinoid-related receptor expression and the tissue distribution as well as the possible interaction between retinoid-related receptor expression and cigarette smoke. METHODOLOGY: Using semiquantitative conventional reverse transcriptase polymerase chain reaction (RT-PCR), quantitative real-time RT-PCR, in situ hybridization and immunostaining, we evaluated the expression and the distribution of retinoic acid receptor alpha, beta, gamma, retinoid X receptor alpha, beta, gamma, thyroid hormone receptor alpha, beta, and alcohol dehydrogenase3 (a well-characterized retinoic acid responsive gene) in malignant and non-malignant tissues obtained from patients with non-small cell lung cancer. We also assessed the response of the bronchial epithelial cell line, BEAS-2B, following exposure to cigarette smoke extract. RESULTS: Compared with non-malignant tissues, malignant tissues had a 45% (P < 0.02) reducion in retinoic acid receptor alpha and beta expression, while other receptors were not affected. The expression of the alcohol dehydrogenase3 gene in malignant tissue was 50% of that observed in non-malignant tissue (P < 0.05). There was a significant correlation between the retinoic acid receptor beta-mRNA level and the alcohol dehydrogenase3-mRNA level (r = 0.52, P < 0.05). In situ hybridization and immunostaining demonstrated that retinoic acid receptor alpha and beta were localized in the airway and alveolar epithelial cells and their expression was diminished to 30% in malignant tissues compared to non-malignant tissues (P < 0.05). Interestingly, cigarette smoke extract decreased retinoic acid receptor beta mRNA in BEAS-2B cells (P < 0.05). CONCLUSIONS: The present findings suggest a possible association of the loss of retinoic acid receptor alpha, beta and alcohol dehydrogenase3 with lung carcinogenesis.

Yields and daily consumption of cigarettes in Japan in 1969-1996.

Cigarette modification trends and the relationship between nicotine yields and consumption in Japan were examined over the 27 years between 1969-1996. Data on cigarette use were obtained from reports published by the government and tobacco manufacturers. Over the study period, there has been a coherent pattern of cigarette modification in Japan. The sales-weighted average yields have declined from 20.7 mg tar and 1.64 mg nicotine/cigarette in 1969 to 8.7 mg tar and 0.72 mg nicotine/cigarette in 1996. On the other hand, the average daily consumption per smoker has continuously increased over the same period. Average nicotine yields and daily cigarette consumption have significant negative correlations among both males and females. This relationship was observed even after controlling for the price changes of cigarettes over time. It is indicated that smokers have compensated for reduced nicotine yields by increasing daily consumption. This may have offset potential benefits of the continuous decline in tar and nicotine yields to smokers' health.

Morphological and biochemical changes following acute unilateral testicular torsion in prepubertal rats.

PURPOSE: The role played by reactive oxygen species in the effects of testicular torsion and torsion/detorsion on ipsilateral and contralateral testis was investigated. MATERIALS AND METHODS: Prepubertal rats were submitted to unilateral testicular torsion alone or followed by detorsion for up to 1 week. Morphology and biochemical parameters (thiobarbituric reactive substances, superoxide dismutase-like and catalase-like activities) were evaluated. RESULTS: Torsion (1 or 2 h) alone induced time-dependent morphological damages that worsened progressively after detorsion in ipsilateral testis but had no effect on contralateral testis. The levels of antioxidants against the superoxide anion and hydrogen peroxide decreased in ipsilateral but not in contralateral testis. However, the level of thiobarbituric reactive substances (an indicator of lipid perioxidation) decreased after testicular torsion or torsion/detorsion in both testes. CONCLUSION: These data emphasize that oxidative stress may play a role in testicular damage caused by torsion/detorsion and that biochemical indicators of oxidative stress are more sensitive than histological techniques in detecting modifications in the contralateral testis.