Dynamics of the Streptococcus gordonii Transcriptome in Response to Medium, Salivary α-Amylase, and Starch.

Streptococcus gordonii, a primary colonizer of the tooth surface, interacts with salivary α-amylase via amylase-binding protein A (AbpA). This enzyme hydrolyzes starch to glucose, maltose, and maltodextrins that can be utilized by various oral bacteria for nutrition. Microarray studies demonstrated that AbpA modulates gene expression in response to amylase, suggesting that the amylase-streptococcal interaction may function in ways other than nutrition. The goal of this study was to explore the role of AbpA in gene regulation through comparative transcriptional profiling of wild-type KS1 and AbpA(-) mutant KS1ΩabpA under various environmental conditions. A portion of the total RNA isolated from mid-log-phase cells grown in 5% CO2 in (i) complex medium with or without amylase, (ii) defined medium (DM) containing 0.8% glucose with/without amylase, and (iii) DM containing 0.2% glucose and amylase with or without starch was reverse transcribed to cDNA and the rest used for RNA sequencing. Changes in the expression of selected genes were validated by quantitative reverse transcription-PCR. Maltodextrin-associated genes, fatty acid synthesis genes and competence genes were differentially expressed in a medium-dependent manner. Genes in another cluster containing a putative histidine kinase/response regulator, peptide methionine sulfoxide reductase, thioredoxin protein, lipoprotein, and cytochrome c-type protein were downregulated in KS1ΩabpA under all of the environmental conditions tested. Thus, AbpA appears to modulate genes associated with maltodextrin utilization/transport and fatty acid synthesis. Importantly, in all growth conditions AbpA was associated with increased expression of a potential two-component signaling system associated with genes involved in reducing oxidative stress, suggesting a role in signal transduction and stress tolerance.

Succinic acid production from xylose mother liquor by recombinant Escherichia coli strain.

Succinic acid (1,4-butanedioic acid) is identified as one of important building-block chemicals. Xylose mother liquor is an abundant industrial residue in xylitol biorefining industry. In this study, xylose mother liquor was utilized to produce succinic acid by recombinant Escherichia coli strain SD121, and the response surface methodology was used to optimize the fermentation media. The optimal conditions of succinic acid fermentation were as follows: 82.62 g L-1 total initial sugars, 42.27 g L-1 MgCO3 and 17.84 g L-1 yeast extract. The maximum production of succinic acid was 52.09 ± 0.21 g L-1 after 84 h with a yield of 0.63 ± 0.03 g g-1 total sugar, approaching the predicted value (53.18 g L-1). It was 1.78-fold of the production of that obtained with the basic medium. This was the first report on succinic acid production from xylose mother liquor by recombinant E. coli strains with media optimization using response surface methodology. This work suggested that the xylose mother liquor could be an alternative substrate for the economical production of succinic acid by recombinant E. coli strains.

Structural effects on persister control by brominated furanones.

Bacterial persister cells are a small population of dormant cells that are tolerant to essentially all antibiotics. Recently, we reported that a quorum sensing (QS) inhibitor, (Z)-4-bromo-5-(bromomethylene)-3-methylfuran-2(5H)-one (BF8), can revert antibiotic tolerance of Pseudomonas aeruginosa persister cells. To better understand this phenomenon, several synthetic brominated furanones with similar structures were compared for their activities in persister control and inhibition of acyl-homoserine lactone (AHL) mediated QS. The results show that some other furanones in addition to BF8 are also AHL QS inhibitors and can revert antibiotic tolerance of P. aeruginosa PAO1 persister cells. However, not all QS inhibiting BFs can revert persistence at growth non-inhibitory concentrations, suggesting that QS inhibition itself is not sufficient for persister control.

Controlling persister cells of Pseudomonas aeruginosa PDO300 by (Z)-4-bromo-5-(bromomethylene)-3-methylfuran-2(5H)-one.

Pseudomonas aeruginosa is a major pathogen causing chronic pulmonary infections; for example, 80% of cystic fibrosis patients get infected by this bacterium as the disease progresses. Such chronic infections are challenging because P. aeruginosa exhibits high-level tolerance to antibiotics by forming biofilms (multicellular structures attached to surfaces), by entering dormancy and forming antibiotic tolerant persister cells, and by conversion to the mucoid phenotype. Recently, we reported that a synthetic quorum sensing inhibitor, (Z)-4-bromo-5-(bromomethylene)-3-methylfuran-2(5H)-one (BF8), can sensitize both planktonic and biofilm-associated persister cells of P. aeruginosa PAO1 to antibiotics at the concentrations non-inhibitory to its growth. In this study, we further characterized the effects of this compound on the mucoid strain P. aeruginosa PDO300. BF8 was found to reduce persistence during the growth of PDO300 and effectively kill the persister cells isolated from PDO300 cultures. In addition to planktonic cells, BF8 was also found to inhibit biofilm formation of PDO300 and reduce associated persistence. These findings broaden the activities of this class of compounds and indicate that BF8 also has other targets in P. aeruginosa in addition to quorum sensing.

(Z)-4-bromo-5-(bromomethylene)-3-methylfuran-2(5H)-one sensitizes Escherichia coli persister cells to antibiotics.

Persisters are a small subpopulation of bacterial cells that are dormant and extremely tolerant to antibiotics. The intrinsic antibiotic tolerance of persisters also facilitates the development of multidrug resistance through acquired mechanisms based on drug resistance genes. In this study, we demonstrate that (Z)-4-bromo-5-(bromomethylene)-3-methylfuran-2(5H)-one (BF8) can reduce persistence during Escherichia coli growth and revert the antibiotic tolerance of its persister cells. The effects of BF8 were more profound when the pH was increased from 6 to 8.5. Although BF8 is a quorum sensing (QS) inhibitor, similar effects were observed for the wild-type E. coli RP437 and its ΔluxS mutant, suggesting that these effects did not occur solely through inhibition of AI-2-mediated QS. In addition to its effects on planktonic persisters, BF8 was also found to disperse RP437 biofilms and to render associated cells more sensitive to ofloxacin. At the doses that are effective against E. coli persister cells, BF8 appeared to be safe to the tested normal mammalian cells in vitro and exhibited no long-term cytotoxicity to normal mouse tissues in vivo. These findings broadened the activities of brominated furanones and shed new light on persister control.

In vivo microcapillary sampling coupled with matrix-assisted laser desorption/ionization fourier transform ion cyclotron resonance mass spectrometry for real-time monitoring of paraquat and diquat in living vegetables.

An in vivo microcapillary sampling (MCS) method coupled with matrix-assisted laser desorption/ionization fourier transform ion cyclotron resonance mass spectrometry (MALDI-FTICR-MS) analysis was utilized to monitor the real-time bipyridine quaternary ammonium herbicides concentrations and assess their uptake and elimination behaviors in living cabbage plants noninvasively. Under optimized conditions, the proposed method for paraquat (PQ) and diquat (DQ) determination showed wide linear ranges (7.81-500 µg/kg), low limits of detection (0.1-0.9 µg/kg), and good reproducibility. In vivo tracking results demonstrated that different absorption behaviors between PQ and DQ existed in living vegetables and DQ was more easily absorbed. Through decay kinetics model fitting, herbicide half-lives were 1.32 and 1.86 days for PQ and DQ, respectively. To summarize, in vivo MCS method provides valuable information on herbicide risks for agricultural production, which is suitable for temporal, spatial, and longitudinal studies in the same living system and multicompartmental studies in the same organism.

Nanobodies Based on a Sandwich Immunoassay for the Detection of Staphylococcal Enterotoxin B Free from Interference by Protein A.

As one of the leading causes of food poisoning, staphylococcal enterotoxins (SEs) secreted by Staphylococcus aureus pose a serious threat to human health. The immunoassay has become the dominant tool used for the rapid detection of harmful bacteria and toxins as a result of its excellent specificity. However, with regard to SEs, staphylococcal protein A (SpA) is likely to bind with the fragment crystallizable (Fc) terminal of the traditional antibody and result in a false positive, limiting the practical application of this method. Therefore, to eliminate the bottleneck problem, the sandwich immunoassay was development by replacing the traditional antibody with a nanobody (Nb) that lacked a Fc terminal. Using 0.5 × 107 colony-forming units, the Nb library was constructed using Bactrian camels immunized with staphylococcal enterotoxin B (SEB) to obtain a paired Nb against SEB with good affinity. A sandwich enzyme-linked immunosorbent assay (ELISA) was developed using one Nb as the capture antibody and a phage-displayed Nb with signal-amplifying properties as the detection antibody. In optimal conditions, the current immunoassay displayed a broad quantitative range from 1 to 512 ng/mL and a 0.3 ng/mL limit of detection. The recovery of spiked milk, milk powder, cheese, and beef ranged from 87.66 to 114.2%. The Nbs-ELISA was not influenced by SpA during the detection of SEB in S. aureus food poisoning. Therefore, the Nb developed here presented the perfect candidates for immunoassay application during SE determination as a result of the complete absence of SpA interference.

Improvement of water quality in the Pearl River Estuary, China: a long-term (2008-2017) case study of temporal-spatial variation, source identification and ecological risk of heavy metals in surface water of Guangzhou.

A series of environmental protective policies have been taken recently in the Pearl River Estuary (PRE) to alleviate water pollution; however, their influence on the reduction of heavy metals in estuarine water has not been known. This study selected Guangzhou as a representative city in the PRE and collected estuarine water monthly from 2008 to 2017 to track the variation of As, Hg, Pb, Cd, Cu, Zn, and Se. During the last decade, the high time-resolved record showed that the concentration of Hg, Pb, Cd, Cu, and Zn in estuarine water reduced by 39.5%, 91.0%, 86.2%, 74.6%, and 97.3%, respectively. However, the concentration of As kept in a stable range (1.89-2.69 µg L-1) and Se (0.17-0.65 µg L-1) increased slightly. The principal component analysis (PCA) and absolute principal component scores-multiple linear regression (APCS-MLR) results suggested that the upstream industrial effluents were major sources for Hg (45.5-92.7%), Pb (47.3-100%), Cd (42.0-90.6%), Cu (85.5-100%), and Zn (100%) and the geogenic source was major origin for As (84.6-98.3%) and Se (0-67.5%). The risk quotient of Hg, Pb, Cd, Cu, and Zn to aquatic organisms largely decreased from 0.03, 0.59, 0.03, 2.06, and 0.26 in 2008 to 0.02, 0.05, 0.006, 0.52, and 0.007 in 2017, respectively. The effective control of heavy metal pollution in the study area can be primarily due to the relocation of hundreds of polluting factories during the last decade.

Short-Chain Modified SiO2 with High Absorption of Organic PCM for Thermal Protection.

Organic phase change materials (PCMs) have great potential in thermal protection applications but they suffer from high volumetric change and easy leakage, which require "leak-proof" packaging materials with low thermal conductivity. Herein, we successfully modify SiO2 through a simple 2-step method consisting of n-hexane activation followed by short-chain alkane silanization. The modified SiO2 (M-SiO2) exhibits superior hydrophobic property while maintaining the intrinsic high porosity of SiO2. The surface modification significantly improves the absorption rate of RT60 in SiO2 by 38%. The M-SiO2/RT60 composite shows high latent heat of 180 J·g-1, low thermal conductivity of 0.178 W·m-1·K-1, and great heat capacity behavior in a high-power thermal circuit with low penetrated heating flow. Our results provide a simple approach for preparing hydrophobic SiO2 with high absorption of organic PCM for thermal protection applications.

[Distribution, Sources, and Health Risk Assessment of PAHs in Water Supply Source Regions of Guangzhou].

Trace polycyclic aromatic hydrocarbons (PAHs) in drinking water sources have significant harmful effects on human health. Water and sediment samples from water source regions of three water treatment plants in Guangzhou were collected and the distributions of 16 kinds of PAHs were analyzed. The human risk of PAHs in the water samples was also evaluated using the Risk Assessment Guidance for Superfund (RAGS) of the United States Environmental Protection Agency (USEPA). The results showed that PAHs in the samples from the three water source regions did not exceed the corresponding standard limit for water quality, and the content of ΣPAHs in suspended solids and sediments was below the medium level. The non-carcinogenic risks (HQ and HI) of PAHs in the water samples were less than 1, and the non-carcinogenic risk was negligible. In addition, Riskingest, Riskdermal, and RiskT for the waters were all in range of 5.53×10-7 to 5.34×10-6, indicating that a carcinogen risk was possible but acceptable. The results of the isomer ratio method indicated that the PAHs in the water sources of the three water plants had a mixed input of pollution, including petroleum discharge, petroleum combustion, and incomplete combustion of wood, coal, and biomass. The total organic carbon (TOC) content of the water and sediment samples was positively correlated with the accumulation and enrichment of low-ring PAHs, and there was a significant positive correlation between PAHs and similar molecules in the sediments. The ΣPAHs in the water and sediment samples were also strongly correlated.

Distribution variation of heavy metals in maricultural sediments and their enrichment, ecological risk and possible source-A case study from Zhelin bay in Southern China.

The study characterized the enrichment, ecological risk and possible source of heavy metals (As, Cd, Cr, Cu, Hg, Ni, Pb and Zn) in sediments from a typical mariculture bay. The concentrations of the metals were analyzed in sediments collected from Zhelin bay. The distribution variation was examined during the past decade, which had an increase tendency till 2011. The enrichment factor and geoaccumulation indices suggested Pb, As, Cu and Zn were minor enrichment and unpolluted to moderately polluted, and Ni at cage mariculture area was moderately-severe enrichment and strongly polluted. This area had medium to high ecological risk, especially at the northwest coastal area of semi-closed bay, with high-medium to high ecological risk. Correlation and principal component analyses indicated that most of heavy metals, especially for As, Pb and Ni, primarily resulted from the combustion of gasoline and diesel fuel and the ship protective layer.

[Distribution Characteristics and Ecological Risk Assessment of Organochlorine Pesticides in Surface Sediments of Zhelin Bay in Guangdong Province, China].

The distribution characteristics and pollution degrees of 20 organochlorine pesticides (OCPs) were investigated in surface sediments from Zhelin Bay south of China and the ecological risk of OCPs and integrated ecological risk at the samples stations in sediments were evaluated by risk quotient (RQ) and sediment quality guideline of NOAA. The possible sources of HCHs and DDTs in sediments were preliminarily studied, and the correlation between OCPs and sediment characteristic parameters was discussed. The concentrations of DDTs in surface sediments were found to be higher than those of other OCPs, especially p,p'-DDD, and the concentrations of DDTs were higher than the limited values of Chinese Marine sediment quality criteria. The total concentration of 20 OCPs in surface sediments ranged from 14.14 to 306.88 ng·g-1, with a mean concentration of 78.37 ng·g-1, and the highest total concentration was at site 8(S8). There were inevitable adverse biological effects and high ecological risk of p,p'-DDD and p,p'-DDT, high integrated ecological risk at S8 and S10. The possible sources of HCHs in surface sediments of Zhelin Bay were mainly from the application of Lindane pesticides. At S1,S2,S10,S11 there was input of DDT into mariculture area of Zhelin Bay, and at the other sites the sources of DDT were from the early residue in sediments or the long-term weathering sediments of using pesticides. There was a very significant positive correlation between HCHs, endosulfan, chlordane and dieldrin in sediments, while there was a negative relation between the above OCPs and TOC, which indicated that the presence of TOC could promote their biodegradation. There was a significant positive correlation between p, p'-DDD and Ca, which revealed that DDT used in the early period was deposited with biological carbonate. Increasing size of sediment particles discouraged the accumulation and enrichment of OCPs in sediments.

[Distribution Characteristics of Heavy Metals in Environmental Samples Around Electroplating Factories and the Health Risk Assessment].

This study aimed to investigate the pollution degree and human health risk of heavy metals in soil and air samples around electroplating factories. Soil, air and waste gas samples were collected to measure 8 heavy metals (As, Cd, Cr, Cu, Hg, Ni, Pb and Zn) in two electroplating factories, located in Baiyun district of Guangzhou city. Geoaccumulation index and USEPA Risk Assessment Guidance for Superfund (RAGS) were respectively carried out. Results showed that concentrations of Hg and Pb in waste gas and Cr in air samples were higher than limits of the corresponding quality standards, and concentrations of Cd, Hg and Zn in soil samples reached the moderate pollution level. The HQ and HI of exposure by heavy metals in air and soil samples were both lower than 1, indicating that there was no non-carcinogen risk. CRAs and CRCr in soil samples were beyond the maximum acceptable level of carcinogen risk (10(-4)), and the contribution rate of CRCr to TCR was over 81%. CRCr, CRNi and TCR in air samples were in range of 10(-6) - 10(-4), indicating there was possibly carcinogen risk but was acceptable risk. CR values for children were higher than adults in soils, but were higher for adults in air samples. Correlation analysis revealed that concentrations of heavy metals in soils were significantly correlated with these in waste gas samples, and PCA data showed pollution sources of Cd, Hg and Zn in soils were different from other metals.

[Characteristics of speciation and evaluation of ecological risk of heavy metals in sewage sludge of Guangzhou].

Contents of heavy metals in different sewage sludges were analyzed and the speciation distribution and bioavailability of heavy metals were investigated, and the risk assessment code (RAC) and toxicity characteristic leaching procedure for solid waste were used to evaluate the potential ecological risk and leaching toxicity risk of heavy metals in sludge samples, respectively. The results showed that contents of Cu, Cr, Pb and Zn were high and presented a great difference by different sources in sewage sludges. Most of heavy metals existed in non-residual fractions and percentages of the mobile fraction (acid soluble fraction) of heavy metals in acidic sludge were higher. According to the results of single extraction, 1 mol x L(-1) NaOAc solution (pH 5.0) and 0.02 mol x L(-1) EDTA + 0.5 mol x L(-1) NH4OAc solution (pH 4.6) were suitable for evaluating bioavailable heavy metals in acidic and alkaline sludge, respectively. Percentages of bioavailable heavy metals were higher with the stronger of sludge acidity. The mobile ability of heavy metals resulted in the high ecological risk of sludge samples, and the bioavailability of heavy metals caused acidic sludges with a very high ecological risk but alkaline sludges with the middle ecological risk. Leaching toxicity risk was very high in sludge samples except domestic sewage sludge. After the removal of bioavailable heavy metals, leaching toxicity risk of sludge samples was still high in spite of its decrease; however, part type of sludges could be implemented landfill disposal.

Reverting antibiotic tolerance of Pseudomonas aeruginosa PAO1 persister cells by (Z)-4-bromo-5-(bromomethylene)-3-methylfuran-2(5H)-one.

BACKGROUND: Bacteria are well known to form dormant persister cells that are tolerant to most antibiotics. Such intrinsic tolerance also facilitates the development of multidrug resistance through acquired mechanisms. Thus persister cells are a promising target for developing more effective methods to control chronic infections and help prevent the development of multidrug-resistant bacteria. However, control of persister cells is still an unmet challenge. METHODOLOGY/PRINCIPAL FINDINGS: We show in this report that (Z)-4-bromo-5-(bromomethylene)-3-methylfuran-2(5H)-one (BF8) can restore the antibiotic susceptibility of Pseudomonas aeruginosa PAO1 persister cells at growth non-inhibitory concentrations. Persister control by BF8 was found to be effective against both planktonic and biofilm cells of P. aeruginosa PAO1. Interestingly, although BF8 is an inhibitor of quorum sensing (QS) in Gram-negative bacteria, the data in this study suggest that the activities of BF8 to revert antibiotic tolerance of P. aeruginosa PAO1 persister cells is not through QS inhibition and may involve other targets. CONCLUSION: BF8 can sensitize P. aeruginosa persister cells to antibiotics.

Quorum sensing inhibitors: a patent overview.

BACKGROUND: Quorum sensing (QS) is a microbial cell-cell signaling system that correlates gene expression with cell population density. It plays important roles in intra-species communications and is also involved in inter-species and microbe-host interactions. Because QS controls a wide spectrum of phenotypes including virulence and biofilm formation, inhibition of QS may provide alternative therapeutic methods for treating microbial infections. OBJECTIVE: To provide an overview of QS inhibitors and their applications. METHODS: Search of databases for patents on QS inhibition submitted between 1999 and 2008. RESULTS/CONCLUSIONS: The reported QS inhibitors include both natural and synthetic agents and can be mainly categorized in three classes: nonpeptide small molecules, peptides and proteins. These inhibitors interrupt QS by repressing signal generation, blocking signal receptors or disrupting QS signals, and provide an alternative approach to controlling microbial pathogenesis.

[Marine SRB community reducing sulfate wastewater in flue gas desulfurization].

An SRB community (SRB-2) was enriched from marine sediment for the treatment of sulfate-rich wastewater of high salinity, and the effect of salinity, temperature, pH value, carbon source, concentration of sulfate and the form of Fe on the activity of SRB-2 was studied. The results show that SRB-2 is a halophilous and moderately mesophilous SRB community. The optimal conditions for its growth are as follows: temperature of 30-40 degrees C and pH value of 7.4-8.3; it can endure 5,200 mg/L SO4(2-) and 60g/L NaCl. Zero-valent Fe can promote the reductive activity of SRB-2, while Fe2+ inhibits that. SEM and optical microscopic measurements indicate many rod-shaped and spiral bacteria on the surface of padding in reactor and black sticky substance composed of rod-shaped bacteria on the bottom of reactor. This sticky substance might be cumulus of culture SRB-2-64 (GenBank accession number: EU167911).