Exopolysaccharide dispelled by calcium hydroxide with volatile vehicles related to bactericidal effect for root canal medication

Objective: Enterococcus faecalis is the dominant microbial species responsible for persistent apical periodontitis with ability to deeply penetrate into the dentin. Exopolysaccharides (EPS) contribute to the pathogenicity and antibiotic resistance of E. faecalis. Our aim was to investigate the antimicrobial activity of calcium hydroxide (CH), camphorated parachlorophenol (CMCP), and chlorhexidine (CHX) against E. faecalis in dentinal tubules. Material and Methods: Decoronated single-canal human teeth and semicylindrical dentin blocks were incubated with E. faecalis for 3 weeks. Samples were randomly assigned to six medication groups for 1 week (n=10 per group): CH + 40% glycerin-water solution (1:1, wt/vol); CMCP; 2% CHX; CH + CMCP (1:1, wt/vol); CH + CMCP (2:3, wt/vol); and saline. Bacterial samples were collected and assayed for colony-forming units. After dentin blocks were split longitudinally, confocal laser scanning microscopy was used to assess the proportion of viable bacteria and EPS production in dentin. Results: CMCP exhibited the best antimicrobial activity, while CH was the least sensitive against E. faecalis (p;0.05). CH combined with CMCP inhibited EPS synthesis by E. faecalis, which sensitized biofilms to antibacterial substances. Moreover, increasing concentrations of CMCP decreased EPS matrix formation, which effectively sensitized biofilms to disinfection agents. Conclusion: The EPS matrix dispelled by CH paste with CMCP may be related to its bactericidal effect; the visualization and analysis of EPS formation and microbial colonization in dentin may be a useful approach to verify medicaments for antimicrobial therapy

Roles of SlETR7, a newly discovered ethylene receptor, in tomato plant and fruit development

Ethylene regulates many aspects of plant growth and development. It is perceived by a family of ethylene receptors (ETRs) that have been well described. However, a full understanding of ETR function is complicated by functional redundancy between the receptor isoforms. Here, we characterize a new ETR, SlETR7, that was revealed by tomato genome sequencing. SlETR7 expression in tomato fruit pericarp increases when the fruit ripens and its expression is synchronized with the expression of SlETR1, SlETR2, and SlETR5 which occurs later in the ripening phase than the increase observed for SlETR3, SlETR4, and SlETR6. We uncovered an error in the SlETR7 sequence as documented in the ITAG 3 versions of the tomato genome which has now been corrected in ITAG 4, and we showed that it belongs to sub-family II. We also showed that SlETR7 specifically binds ethylene. Overexpression (OE) of SlETR7 resulted in earlier flowering, shorter plants, and smaller fruit than wild type. Knock-out (KO) mutants of SlETR7 produced more ethylene at breaker (Br) and Br + 2 days stages compared to wild type (WT), but there were no other obvious changes in the plant and fruit in these mutant lines. We observed that expression of the other SlETRs is upregulated in fruit of SlETR7 KO mutants, which may explain the absence of obvious ripening phenotypes. Globally, these results show that SlETR7 is a functional ethylene receptor. More work is needed to better understand its specific roles related to the six other tomato ETRs

Pathway and kinetics of cyhalothrin biodegradation by Bacillus thuringiensis strain ZS-19

10.1038/srep08784Scientific Reports

Browning of Pig White Preadipocytes by Co-Overexpressing Pig PGC-1α and Mice UCP1

Background/Aims: Brown adipose tissue (BAT) is critical for mammals’ survival in the cold environment. BAT-dependent non-shivering thermogenesis is attributed to uncoupling protein 1 (UCP1)’s disengagement of oxidative phosphorylation from ATP synthesis and dissipates energy as heat. Thus individuals with a substantial amount of BAT are better equipped during cold stress and less likely to become obese. Recently, our laboratory has shown pig adipocytes have no UCP1 protein. The inability of newborn piglets to generate heat contributed to its high death rate. Repairing the genetic defect of UCP1 in pig adipocytes has implications in defending against cold for piglets and developing an alternative treatment for human obesity. Methods: Q-PCR, western blotting (WB) and oxygen consumption measurement were used to enable functional UCP1 protein in preadipocytes. Immunoprecipitation (IP), chromatin immunoprecipitation (CHIP), and dual-luciferase reporter assay system were used to clarify the thermogenesis mechanism of functional UCP1. Results: Only co-overexpressing mice UCP1 and pig PGC-1α increased not only the mitochondrial number but also the uncoupled respiration rate in the transfected pig adipocytes. The functional mice UCP1 increased the pig PGC-1α activity through the AMPK-SIRT1 pathway. The active form PGC-1α interacted with transcription factors Lhx8, Zic1, ERRα, and PPARα to regulate the expression of mitochondrial energy metabolism and adipocytes browning-related genes. Conclusion: Our data suggest a model in which pig PGC-1α and mice UCP1 work collaboratively to restore uncoupling respiration in pig preadipocytes. These results have great implications for piglet survival and developing an alternative treatment for human obesity in the future

Scaling from single-point sap velocity measurements to stand transpiration in a multispecies deciduous forest: Uncertainty sources, stand structure effect, and future scenarios

9 páginas.-- 5 figuras.-- 2 tablas.-- 58 referencias[EN] A major challenge in studies estimating stand water use in mixed-species forests is how to effectively scale data from individual trees to the stand. This is the case for forest ecosystems in the northeastern USA where differences in water use among species and across different size classes have not been extensively studied, despite their relevance for a wide range of ecosystem services. Our objectives were to assess the importance of different sources of variability on transpiration upscaling and explore the potential impacts of future shifts in species composition on the forest water budget. We measured sap velocity in five tree species (Fagus grandifolia Ehrh., Acer rubrum L., Acer saccharum Marsh., Betula alleghaniensis Britton, and Betula papyrifera Marsh.) in a mature stand and a young stand in New Hampshire, USA. Our results showed that the greatest potential source of error was radial variability and that tree size was more important than species in determining sap velocity. Total sapwood area was demonstrated to exert a strong controlling influence on transpiration, varying depending on tree size and species. We conclude that the effect of potential species shifts on transpiration will depend on the sap velocity, determined not only by radial variation and tree size, but also by the sapwood area distribution in the stand.[FR] Les études dont le but est d'estimer l'utilisation de l'eau a` l'échelle du peuplement dans les forêts mélangées font face a` un défi majeur : comment passer efficacement de l'échelle des arbres individuels a` l'échelle du peuplement. C'est le cas pour les écosystèmes forestiers dans le nord-est des États-Unis où les différences dans l'utilisation de l'eau entre les espèces et parmi les différentes catégories de taille n'ont pas fait l'objet d'études approfondies malgré leur pertinence pour une vaste gamme de services de l'écosystème. Nos objectifs consistaient a` évaluer l'importance des différentes sources de variation sur l'extrapolation de la transpiration et a` explorer les impacts potentiels des changements futurs dans la composition en espèces sur le bilan hydrique de la forêt. Nous avons mesuré la vitesse de la sève chez cinq espèces d'arbre (Fagus grandifolia Ehrh., Acer rubrum L., Acer saccharum Marsh., Betula alleghaniensis Britton et Betula papyrifera Marsh.) dans un peuplement mature et dans un jeune peuplement au New Hampshire (É.-U.). Nos résultats ont montré que la plus grande source potentielle d'erreur était la variation radiale et que la vitesse de la sève était davantage déterminée par la taille des arbres que par l'espèce. La surface totale de bois d'aubier avait un effet très déterminant sur la transpiration qui variait selon la taille et l'espèce d'arbre. Nous concluons que l'effet des changements potentiels dans la composition en espèces sur la transpiration dépendra de la vitesse de la sève qui est principalement déterminée par la variation radiale et la taille des arbres mais aussi de la distribution de la surface de bois d'aubier dans le peuplement.This work was funded by the University of New Hampshire and the New Hampshire Agricultural Experiment Station. The Bartlett Experimental Forest is operated by the USDA Forest Service Northern Research Station. S. Mcgraw, P. Pellissier, C. Breton, S. Alvarado-Barrientos, R. Snyder, and Z. Aldag assisted in the field and in the lab. The 2011 stand inventory was led by S. Goswami. Tree heights were measured and compiled by C. Blodgett, T. Fahey, and L. Liu. A. Richardson shared meteorology and solar radiation data from the Bartlett Amerflux tower. The stands used in this experiment are maintained and monitored by the MELNHE project under the direction of R. Yanai and M. Fisk, with funding from NSF grants DEB 0235650 and DEB 0949324Peer reviewe

Purification and Characterization of a Novel Chlorpyrifos Hydrolase from Cladosporium cladosporioides Hu-01

Chlorpyrifos is of great environmental concern due to its widespread use in the past several decades and its potential toxic effects on human health. Thus, the degradation study of chlorpyrifos has become increasing important in recent years. A fungus capable of using chlorpyrifos as the sole carbon source was isolated from organophosphate-contaminated soil and characterized as Cladosporium cladosporioides Hu-01 (collection number: CCTCC M 20711). A novel chlorpyrifos hydrolase from cell extract was purified 35.6-fold to apparent homogeneity with 38.5% overall recovery by ammoniumsulfate precipitation, gel filtration chromatography and anion-exchange chromatography. It is a monomeric structure with a molecular mass of 38.3 kDa. The pI value was estimated to be 5.2. The optimal pH and temperature of the purified enzyme were 6.5 and 40°C, respectively. No cofactors were required for the chlorpyrifos-hydrolysis activity. The enzyme was strongly inhibited by Hg2+, Fe3+, DTT, β-mercaptoethanol and SDS, whereas slight inhibitory effects (5–10% inhibition) were observed in the presence of Mn2+, Zn2+, Cu2+, Mg2+, and EDTA. The purified enzyme hydrolyzed various organophosphorus insecticides with P-O and P-S bond. Chlorpyrifos was the preferred substrate. The Km and Vmax values of the enzyme for chlorpyrifos were 6.7974 μM and 2.6473 μmol·min−1, respectively. Both NH2-terminal sequencing and matrix-assisted laser desorption/ionization time-of-flight/time-of-flight mass spectrometer (MALDI-TOF-MS) identified an amino acid sequence MEPDGELSALTQGANS, which shared no similarity with any reported organophosphate-hydrolyzing enzymes. These results suggested that the purified enzyme was a novel hydrolase and might conceivably be developed to fulfill the practical requirements to enable its use in situ for detoxification of chlorpyrifos. Finally, this is the first described chlorpyrifos hydrolase from fungus

Microbial Detoxification of Bifenthrin by a Novel Yeast and Its Potential for Contaminated Soils Treatment

Bifenthrin is one the most widespread pollutants and has caused potential effect on aquatic life and human health, yet little is known about microbial degradation in contaminated regions. A novel yeast strain ZS-02, isolated from activated sludge and identified as Candida pelliculosa based on morphology, API test and 18S rDNA gene analysis, was found highly effective in degrading bifenthrin over a wide range of temperatures (20–40°C) and pH (5–9). On the basis of response surface methodology (RSM), the optimal degradation conditions were determined to be 32.3°C and pH 7.2. Under these conditions, the yeast completely metabolized bifenthrin (50 mg·L−1) within 8 days. This strain utilized bifenthrin as the sole carbon source for growth as well as co-metabolized it in the presence of glucose, and tolerated concentrations as high as 600 mg·L−1 with a qmax, Ks and Ki of 1.7015 day−1, 86.2259 mg·L−1 and 187.2340 mg·L−1, respectively. The yeast first degraded bifenthrin by hydrolysis of the carboxylester linkage to produce cyclopropanecarboxylic acid and 2-methyl-3-biphenylyl methanol. Subsequently, 2-methyl-3-biphenylyl methanol was further transformed by biphenyl cleavage to form 4-trifluoromethoxy phenol, 2-chloro-6-fluoro benzylalcohol, and 3,5-dimethoxy phenol, resulting in its detoxification. Eventually, no persistent accumulative product was detected by gas chromatopraphy-mass spectrometry (GC-MS) analysis. This is the first report of a novel pathway of degradation of bifenthrin by hydrolysis of ester linkage and cleavage of biphenyl in a microorganism. Furthermore, strain ZS-02 degraded a variety of pyrethroids including bifenthrin, cyfluthrin, deltamethrin, fenvalerate, cypermethrin, and fenpropathrin. In different contaminated soils introduced with strain ZS-02, 65–75% of the 50 mg·kg−1 bifenthrin was eliminated within 10 days, suggesting the yeast could be a promising candidate for remediation of environments affected by bifenthrin. Finally, this is the first described yeast capable of degrading bifenthrin