Evaluation of a creosote-based medium for the growth and preparation of a PAH-degrading bacterial community for bioaugmentation

Creosote was evaluated as an inexpensive carbon source for growing inocula of a polycyclic aromatic hydrocarbon (PAH)-degrading bacterial community (community five). Creosote was a poor growth substrate when provided as sole carbon source in a basal salts solution (BSM). Alternatively, peptone, yeast extract or glucose in BSM supported high growth rates, but community five could not subsequently degrade pyrene. A combination of creosote and yeast extract in BSM (CYEM) supported growth and maintained the pyrene-degrading capacity of community five. Optimum pyrene-degrading activity occurred when the inocula were grown in creosote and yeast extract concentrations of 2 ml L⁻¹ and 1 g L⁻¹ respectively: concentrations outside these values resulted in either low biomass yields or loss of PAH-degrading activity. CYEM-grown community five inocula degraded 250 mg L⁻¹ of pyrene in BSM at a rate comparable to cultures inoculated with community five grown in BSM-pyrene. However, the CYEM-grown community showed a 40% lower rate of PAH degradation in a synthetic PAH mixture compared with pyrene-grown cells and there was an increase in the lag period before the onset of PAH degradation. This appears to reflect a weaker induction of PAH catabolism by CYEM compared to BSM-pyrene. Journal of Industrial Microbiology & Biotechnology (2000) 24, 277–284. Received 24 August 1999/ Accepted in revised form 20 January 2000     

高剂量生长抑素、奥美拉唑联合止血芳酸治疗急性上消化道出血合并凝血功能障碍患者的临床效果

目的:探讨高剂量生长抑素、奥美拉唑联合止血芳酸治疗急性上消化道出血合并凝血功能障碍患者的临床效果及安全性。方法:选择我院2014年1月~2017年12月收治的92例急性上消化出血合并凝血功能障碍的患者,并按随机数表法将其分为对照组和研究组。对照组予以常规剂量生长抑素、奥美拉唑联合止血芳酸治疗,研究组予以高剂量生长抑素治疗,其余奥美拉唑及止血芳酸用法同对照组。治疗后,比较两组的临床疗效、止血情况、住院时间,治疗前后血常规指标、凝血功能的变化及并发症的发生情况。结果:治疗后,研究组总有效率明显高于对照组[91.30%vs.74.42%](P0.05),而平均止血时间、再止血率及住院时间均明显短于对照组(P0.05);两组白细胞计数(WBC)、部分活化凝血酶原时间(APTT)及凝血酶原时间(PT)均较治疗前明显下降,血红蛋白(Hb)、红细胞计数(RBC)、红细胞压积(Hct)及血小板计数(PLT)均较治疗前明显上升,且研究组以上指标变化较对照组更明显(P0.05)。两组并发症的发生率比较差异均无统计学意义(P0.05)。结论:高剂量生长抑素、奥美拉唑联合止血芳酸治疗急性上消化道出血合并凝血功能障碍的效果明显优于常规剂量生长抑素、奥美拉唑联合止血芳酸治疗,其能够更有效缩短止血时间,避免再出血,且未增加药物不良反应,安全性高。     

AMF和PGPR联合修复菲和芘污染土壤的效应

丛枝菌根真菌（arbuscular mycorrhizal fungi,AMF）与根围促生细菌（plant growth-promoting rhizobacteria,PGPR）联合降解有毒有机物、修复污染土壤和促进植物生长的作用倍受关注。本试验旨在探究AMF与PGPR联合降解土壤中菲和芘的效应,以菲和芘1:1混合处理浓度各0、50mg/kg、100mg/kg和150mg/kg下对高羊茅Festuca elata接种AMF根内根孢囊霉Rhizophagus intraradices（Ri）、变形球囊霉Glomus versiforme（Gv）、PGPR荧光假单胞菌Pseudomonas fluorescens Ps2-6、芽孢杆菌Bacillus velezensis Ps3-2、Ri+Ps2-6、Ri+Ps3-2、Gv+Ps2-6、Gv+Ps3-2和不接种对照共36个处理。结果表明,供试AMF增加了PGPR的定殖数量;接种PGPR则显著提高AMF的侵染率。AMF、PGPR或AMF+PGPR处理均显著降低土壤中菲和芘含量,促进植物对土壤中菲和芘的吸收,显著提高高羊茅根系和叶片内的菲和芘含量。在土壤中菲和芘100mg/kg和150mg/kg水平下,Gv与Ps2-6及Ri与Ps2-6能相互促进对土壤中菲和芘的去除效应,其中接种Gv+Ps2-6组合处理的去除率最高,达到95%-98%,土壤中多酚氧化酶、脱氢酶和过氧化氢酶活性显著高于单接种处理和不接种对照,而酸性磷酸酶活性变化则表现为相反趋势。其中以Gv+Ps2-6组合处理的多酚氧化酶活性最高,为0.17mg/g,是不接种对照的1.9倍;脱氢酶和过氧化氢酶活性分别达到1.32µg/(g·h)和1.81mL/g;酸性磷酸酶活性则比不接种对照土壤降低27%-45%;易提取球囊霉素相关土壤蛋白含量和总球囊霉素相关土壤蛋白含量分别是不接种对照的1.6倍和1.5倍。     

Electron Paramagnetic Resonance and Spin Trapping Study of Radicals Formed During Reaction of Aromatic Amines with isoAmyl Nitrite Under Aprotic Conditions

Diazotization of primary aromatic amines with isoamyl nitrite in benzene at room temperature was studied employing EPR and spin trapping techniques. Nitrosodurene (ND). 2-methyl-2-nitrosopropane (MNP). and 5,5-dimethyl-pyrroline N-oxide (DMPO) were used as spin trapping agents. Aryl radicals were detected employing ND and MNP. Using DMPO as a spin trap most of the amines produced EPR spectra ascribed to adducts with aniline-type radicals (N-centred radicals). The assignments were verified using ¹⁵JN-labeled anilines. Similar spectra of DMPO adducts were recorded from amines treated with benzoyl peroxide or benzophenone plus UV. Possible mechanisms of formation of these adducts (radical trapping versus nucleophilic addition to DMPO followed by oxidation) during treatment of the amines with isoamyl nitrite are discussed.     

Production of 2-Phenylethanol in Roses as the Dominant Floral Scent Compound from L-Phenylalanine by Two Key Enzymes,a PLP-Dependent Decarboxylase and a Phenylacetaldehyde Reductase

We investigated the biosynthetic pathway for 2-phenylethanol, the dominant floral scent compound in roses, using enzyme assays. L-[²H₈] Phenylalanine was converted to [²H₈] phenylacetaldehyde and [²H₈]-2-phenylethanol by two enzymes derived from the flower petals of R. ‘Hoh-Jun,’ these being identified as pyridoxal-5′-phosphate-dependent L-aromatic amino acid decarboxylase (AADC) and phenylacetaldehyde reductase (PAR). The activity of rose petal AADC to yield phenylacetaldehyde was nine times higher toward L-phenylalanine than toward its D-isomer, and this conversion was not inhibited by iproniazid, a specific inhibitor of monoamine oxidase. Under aerobic conditions, rose petal AADC stoichiometrically produced NH₃ together with phenylacetaldehyde during the course of decarboxylation and oxidation, followed by the hydrolysis of L-phenylalanine. Phenylacetaldehyde was subsequently converted to 2-phenylethanol by the action of PAR. PAR showed specificity toward several volatile aldehydes.     

Molecular dynamics simulations of the aggregation behaviour of overlapped graphene sheets in linear aliphatic hydrocarbons

Molecular dynamics simulations were used to investigate the aggregation of two partially overlapped graphene sheets in hexane, dodecane and eicosane. When partially overlapped graphene sheets are adjacent to one another, they will expel the adsorbed layers of the solvent molecules on the graphene surface, and the amount of overlap will increase. When the overlapped regions of the graphene sheets are separated by solvent molecules, they cannot expel the adsorption layers between them, and so the sheets remain separated. The driving force for aggregation is the van der Waals interaction between the two graphene sheets, while the van der Waals interaction between the graphene sheets and the solvent molecules inhibits graphene aggregation. The diffusion rate of the hydrocarbon molecules with shorter chain lengths is higher. Thus, they diffuse faster during graphene aggregation, which leads to a higher rate of graphene overlapping in the shorter hydrocarbons. This work provides useful insights into graphene aggregation in linear hydrocarbon solvents of varying lengths at the nanoscale.     

干酪乳杆菌arat基因的敲除及其对苯乳酸合成代谢的影响

苯乳酸（PLA）作为一种新型的广谱抑菌物质,在食品行业具有巨大的发展潜力,作为乳酸菌的天然代谢产物之一,对其代谢机理的研究具有非常重要的意义。通过CRISPR/Cas9基因编辑系统对一株干酪乳杆菌（Lactobacillus casei）1.8727的芳香族氨基转移酶基因arat进行敲除,得到一株arat缺失菌株（L. casei）1.8727Δarat。通过HPLC方法分析其代谢产物,发现发酵72 h时,PLA产量比出发菌株提高约66.7%,而苯丙氨酸（Phe）比出发菌株提高约57.8%。首次在L. casei 1.8727中成功应用基因编辑手段敲除了arat基因,研究表明该菌株具有自身合成代谢Phe的能力,arat作为PLA代谢途径的关键酶基因,其缺失并未使PLA产量下调,而是促进了PLA与Phe的合成代谢,证明干酪乳杆菌中可能存在其他的代偿途径,arat基因缺失所造成的代谢流的改变最终造成PLA与Phe产量的提高,同时PLA的合成代谢可能涉及多个基因的参与,是一个复杂的代谢网络。研究结果对于深入研究PLA的合成代谢机制具有重要意义,为后续PLA合成代谢途径提供了新的思路及方法。     

Bioremediation of Polyaromatic Hydrocarbon-Contaminated Sediments in Aerated Bioslurry Reactors

Treatment of dredged sediments contaminated by polyaromatic hydrocarbons (PAHs) is a significant problem in the New York/New Jersey (NY/NJ) Harbor. 0.5 m³-scale slurry-phase bioreactors were used to determine whether bioaugmentation with a PAH-degradative bacterial consortium, or with the salt marsh grass S. alterniflora, could enhance the biodegradation of PAHs added to dredged estuarine sediments from the NY/NJ Harbor. The results were compared to biodegradation effected by the indigenous sediment microbial community. Sediments were diluted 1:1 in tap water and spiked to a final concentration of 20 mg/kg dry weight sediment of phenanthrene, anthracene, acenaphthene, fluorene, fluoranthene, and pyrene. The sediment slurry was then continuously sparged with air over 3 months. In all bioreactors a rapid reduction of greater than 95% of the initial phenanthrene, acenaphthene, and fluorene occurred within 14 days. Pyrene and fluoranthene reductions of 70 to 90% were achieved by day 77 of treatment. Anthracene was more recalcitrant and reductions ranged from 30 to 85%. Separate experiments showed that the sediment microbial communities mineralized ¹⁴C-pyrene and ¹⁴C-phenanthrene. PAH degradation, and the number of phenanthrene-degrading bacteria, were not enhanced by microbial or plant bioaugmentation. These data demonstrate that bioaugmentation is not required to effect efficient remediation of PAH-contaminated dredged sediments in slurry-phase bioreactors.     

Effects of geometric isomerism in dinuclear antitumor platinum complexes on their interactions with N-acetyl-L-methionine

In this study, the reactions of N-acetyl-L-methionine (AcMet) with [{trans-PtCl(NH₃)₂}₂-μ-H₂N(CH₂)₆NH₂](NO₃)₂ (BBR3005: 1,1/t,t 1) and its cis analog [{cis-PtCl(NH₃)₂}₂-μ-{H₂N(CH₂)₆NH₂}]Cl₂ (1,1/c,c 2) were analyzed to determine the rate and reaction profile of chloride substitution by methionine sulfur. The reactions were studied in PBS buffer at 37°C by a combination of multinuclear (¹⁹⁵Pt, {¹H-¹⁵N} HSQC) magnetic resonance (NMR) spectroscopy and electrospray ionization time of flight mass spectrometry (ESITOFMS). The diamine linker of the 1,1/t,t trans complex was released as a result of the trans influence of the coordinated sulfur atom, producing trans-[PtCl(AcMet)(NH₃)₂]⁺ (III) and trans-[Pt(AcMet)₂(NH₃)₂]²⁺ (IV). In contrast the cis geometry of the dinuclear compound maintained the diamine bridge intact and a number of novel dinuclear platinum compounds obtained by stepwise substitution of sulfur on both platinum centers were identified. These include (charges omitted for clarity): [{cis-PtCl(NH₃)₂}-μ-NH₂(CH₂)₆NH₂-{cis-Pt(AcMet)(NH₃)₂}] (V); [{cis-Pt(AcMet)(NH₃)₂}₂-μ-NH₂(CH₂)₆NH₂] (VI); [{cis-PtCl(NH₃)₂}-μ-NH₂(CH₂)₆NH₂-{PtCl(AcMet)NH₃] (VII); [{PtCl(AcMet)(NH₃)}₂-μ-NH₂(CH₂)₆NH₂] (VIII); [{trans-Pt(AcMet)₂(NH₃)}-μ-NH₂(CH₂)₆NH₂-{PtCl(AcMet)(NH₃)] (IX) and the fully substituted [{trans-Pt(AcMet)₂(NH₃)}₂-μ-{NH₂(CH₂)₆NH₂] (X). For both compounds the reactions with methionine were slower than those with glutathione (Inorg Chem 2003, 42:5498–5506). Further, the 1,1/c,c geometry resulted in slower reaction than the trans isomer, because of steric hindrance of the bridge, as observed previously in reactions with DNA and model nucleotides.     

Development of short and highly potent self‐assembling elastin‐derived pentapeptide repeats containing aromatic amino acid residues

Tropoelastin is the primary component of elastin, which forms the elastic fibers that make up connective tissues. The hydrophobic domains of tropoelastin are thought to mediate the self‐assembly of elastin into fibers, and the temperature‐mediated self‐assembly (coacervation) of one such repetitive peptide sequence (VPGVG) has been utilized in various bio‐applications. To elucidate a mechanism for coacervation activity enhancement and to develop more potent coacervatable elastin‐derived peptides, we synthesized two series of peptide analogs containing an aromatic amino acid, Trp or Tyr, in addition to Phe‐containing analogs and tested their functional characteristics. Thus, position 1 of the hydrophobic pentapeptide repeat of elastin (X¹P²G³V⁴G⁵) was substituted by Trp or Tyr. Eventually, we acquired a novel, short Trp‐containing elastin‐derived peptide analog (WPGVG)₃ with potent coacervation ability. From the results obtained during this process, we determined the importance of aromaticity and hydrophobicity for the coacervation potency of elastin‐derived peptide analogs. Generally, however, the production of long‐chain synthetic polypeptides in quantities sufficient for commercial use remain cost‐prohibitive. Therefore, the identification of (WPGVG)₃, which is a 15‐mer short peptide consisting simply of five natural amino acids and shows temperature‐dependent self‐assembly activity, might serve as a foundation for the development of various kinds of biomaterials. Copyright © 2015 European Peptide Society and John Wiley & Sons, Ltd.