Evidence for an additional disulfide reduction pathway in Escherichia coli

An Escherichia coli cell-free protein synthesis cell extract has been created that lacks all known cytoplasmic disulfide reduction pathways but still retains significant reductase activity. Oxidized glutathione was partially stabilized by deleting the gene for glutathione reductase. To avoid previously reported AhpC mutations, thioredoxin reductase was only removed after extract preparation. The trxB gene was extended to encode a hemagglutinin tag so that TrxB could be removed by affinity adsorption. However, significant glutathione reductase activity remained. The unknown glutathione reductase pathway is disabled by iodoacetamide, is inhibited by NADH, and appears to use NADPH as an electron source.     

Persistence and biodegradation of monoethanolamine and 2-propanolamine at an abandoned industrial site

Soil and groundwater samples were collected at the site of a former chemical processing plant in areas impacted by accidental releases of MEA (monoethanolamine) and IPA (2-propanolamine or isopropanolamine). Although their use had ceased ca. 10 years before sample collection, soils collected at contamination sites had MEA concentrations ranging from ca. 400 to 3000 mg/kg and IPA concentrations from ca. 30 to 120 mg/kg. Even though alkanolamines are miscible in water, transport to groundwater was slow, apparently because they are present in soil as bound cations. Only one groundwater sample (near the most highly contaminated soil)from wells directly adjacentto and down-gradient from the contaminated soils had detectable MEA, and none had detectable IPA. However, ammonia was found in the soil samples collected in the MEA-contaminated areas (ca. 500-1400 mg/kg) and the groundwater (80-120 mg/L), as would be consistent with bacterial degradation of MEA to ammonia, followed by transport of ammonia into the groundwater. Counts for bacteria capable of using MEA or IPA as a sole carbon source were ca. 5 x 106 and 1 x 106 (respectively) per gram in uncontaminated site soil, but no such organisms were found in highly contaminated soils. Similarly, bacterial degradation of MEA in slurries of highly contaminated soils was slow, with ca. 8-20 days required for half of the initial concentrations of MEA to be degraded at 20 degrees C and 30-60 days at 10 degrees C. In contrast, bacterial degradation studies using uncontaminated site soils spiked with ca. 1300 mg/L either MEA or IPA showed very rapid degradation of both compounds,with more than 99% degradation occurring in less than 3 days with quantitative conversion to ammonia, followed by slower conversion to nitrite and nitrate. The results obtained in the site soils, the groundwater samples, and from the biodegradation studies demonstrate that MEA and IPA can persist for decades on soil at high (hundreds of mg/kg) concentrations without significant migration into groundwater, despite the fact that they are miscible in water. Since MEA and IPA exist primarily as cations at the pH of site soils, their persistence apparently results from strong binding to soil, as well as inhibition of natural bioremediation in highly contaminated field soils.     

Prediction of principal metal-binding solid phases in estuarine sediments from color image analysis

A novel application of image analysis technology to the semiquantitative determination of the strong metal-binding phases of hydrous ferric oxides and acid-volatile sulfides in muddy estuarine sediments is presented. Sediment cores were split in the laboratory, photographed in cross-section, and subsampled for solid-phase analyses. The images were processed using commercially available software packages. Concentrations of FeOOH and AVS were found to be linearly related to elementary color parameters (saturation and intensity, respectively) at corresponding points on the images. Spatially resolved information on metal-binding capacity can thereby be estimated from such images; selected depth profiles are generated as examples.     

Growth characteristics of calves fed an intensified milk replacer regimen with additional lactoferrin

The objective of this study was to evaluate the effect of lactoferrin addition to milk replacer varying in crude protein (CP) on dry matter intake, growth, and days medicated. Thirty-four Holstein heifer calves were assigned to 4 treatments in a 2 × 2 factorial arrangement of treatments in a randomized complete block design. Treatments were as follows: 562 g daily of a nonmedicated conventional milk replacer (20% CP:20% fat) feeding regimen with or without 1 g of supplemental bovine lactoferrin (n = 9 for both treatments) or a nonmedicated intensified milk replacer feeding regimen (28% CP:20% fat) fed on a metabolizable energy basis (0.2 Mcal/kg BW^0.75) from d 2 to 9, and at 0.27 Mcal/kg BW^0.75 from d 10 to 42 with or without 1g supplemental bovine lactoferrin (n = 8 for both treatments). Calves were fed pelleted starter (25% CP) in 227.5-g increments beginning on d 2 and had free access to water. Calves remained on the study for 14 d postweaning. Dry matter intake was determined daily. Growth measurements were taken weekly. Blood samples were taken twice weekly for determination of blood urea N. On d 10 of life, calves were subjected to a xylose challenge. Calves on conventional treatments ate more starter preweaning, during weaning, and postweaning. Preweaning, intensively fed calves had higher dry matter intakes. Weights of intensified-fed calves were greater at weaning. Intensified milk replacer-fed calves had greater average daily gain preweaning and overall and higher gain:feed ratios preweaning, but conventionally fed calves had higher gain:feed ratios during weaning. Intensified milk replacer-fed calves had greater hip heights during weaning and postweaning and greater heart girths preweaning, weaning, and postweaning. Days medicated were greater preweaning and overall for intensified-fed calves. There were no differences among treatments for xylose absorption. Calves on conventional treatments had increased blood urea nitrogen concentrations preweaning. There were no effects of lactoferrin on any experimental variable. Intensified milk replacer-fed calves consumed less starter but had higher average daily gains overall and larger frames and greater BW than conventionally fed calves. An intensified milk replacer feeding regimen promotes faster growth during the preweaning period when compared with calves fed conventional treatments, but supplemental bovine lactoferrin was not beneficial under these experimental conditions.     

湖南省某砷制品厂场址土壤修复治理工程及场地环境现状的调查

由于污染事故,某砷制品厂关闭,遗留下的污染源问题并未彻底消除,当年污染的场地也未进行风险管控或修复.因此开展本次场地环境现状调查,旨在为项目所在地下一步的环境修复或风险管控提供意见和参考.     

高层住宅现浇混凝土外墙外保温体系的施工实践

钢筋混凝土与聚苯板复合外墙外保温技术及其施工要点。     

压力棒隔距块在棉纺中的应用

为了探讨在棉纺生产中压力棒隔距块对成纱质量的影响,分析了压力棒隔距块的原理,并在FA506型环锭细纱机上加装不同型号的压力棒隔距块分别纺制出CJ9.7 tex、CJ7.3 tex纱线,测试纱线的毛羽、强力、条干等各项指标并进行对比分析。结果表明,在棉纺中使用压力棒隔距块后,成纱条干有所改善,成纱强力变化不大,成纱毛羽的变化也不明显。     

高产梳棉机附加分梳件的分析

为确保在梳棉机产量提高的同时,具有足够的梳理度和除杂效率,采用扩大梳理面,加装分梳板、固定盖板、双刺辊、三刺辊等措施来实现。系统论述了这些措施,指出各种附加分梳件与给棉板、刺辊、锡林、盖板的主分梳作用间存在交互作用、相互影响、相互替代,生条、成纱质量的改善是分梳板、固定盖板、回转盖板、锡林、刺辊等综合作用的结果。根据转移曲线得出只有提高梳棉机一次梳理的能力,才能极大地改善棉网、成纱质量,加装各种附加分梳件提高了一次梳理能力。试验表明,梳针式优于锯齿式,应以梳针代替锯齿,提高分梳件的梳理质量。     

Source apportionment of PM2.5 at an urban IMPROVE site in Seattle, Washington

The multivariate receptor models Positive Matrix Factorization (PMF) and Unmix were used along with the EPA's Chemical Mass Balance model to deduce the sources of PM2.5 at a centrally located urban site in Seattle, WA. A total of 289 filter samples were obtained with an IMPROVE sampler from 1996 through 1999 and were analyzed for 31 particulate elements including temperature-resolved fractions of the particulate organic and elemental carbon. All three receptor models predicted that the major sources of PM2.5 were vegetative burning (including wood stoves), mobile sources, and secondary particle formation with lesser contributions from resuspended soil and sea spray. The PMF and Unmix models were able to resolve a fuel oil combustion source as well as distinguish between diesel emissions and other mobile sources. In addition, the average source contribution estimates via PMF and Unmix agreed well with an existing emissions inventory. Using the temperature-resolved organic and elemental carbon fractions provided in the IMPROVE protocol, rather than the total organic and elemental carbon, allowed the Unmix model to separate diesel from other mobile sources. The PMF model was able to do this without the additional carbon species, relying on selected trace elements to distinguish the various combustion sources.     

Secretion of a variant of human single-chain urokinase-type plasminogen activator without an N-glycosylation site in the methylotrophic yeast,Pichia pastoris and characterization of the secreted product

Human single-chain urokinase-type plasminogen activator without an N-glycosylation site (scu-PA-Q³⁰²) was produced in the methylotrophic yeast, Pichia pastoris using the shortened prepeptide sequence of a fungal aspartic proteinase, Mucor pusillus rennin (MPR). The level of urokinase-type plasminogen activator (u-PA) immunoreactive material in YPM medium was 0·47 mg/l; however, most of the secreted product had been processed to smaller polypeptides. The N-terminal amino acid sequence of major species was identical to that of the low molecular weight two-chain u-PA. Some approaches to minimizing the proteolysis of scu-PA-Q³⁰² were attempted. Addition of Triton X-100, l-arginine and ammonium phosphate to the YPM medium minimized the proteolysis of scu-PA-Q³⁰² and increased the yield of immunoreactive material to approximately 5 mg/l. Use of proteinase A- or proteinase B-deficient strains of yeast did not reduce the degradation. Co-expression of scu-PA-Q³⁰² and urinary trypsin inhibitor resulted in partial reduction of the major species of proteolysis. Scu-PA-Q³⁰² was purified from the culture supernatant of the improved medium by two successive chromatographies on Phenyl-Sepharose and S-Sepharose. The purified protein had a molecular weight of 47 kDa. It did not contain detectable N-linked oligosaccharides, but contained O-linked oligosaccharides attached to the light chain. N-terminal amino acid sequencing of the purified preparation showed that the shortened prepeptide sequence of MPR was correctly processed by the Pichia yeast. Scu-PA-Q³⁰² closely resembles natural scu-PA with respect to its enzymatic activity against the chromogenic substrate S-2444 and its in vitro fibrinolytic properties.     

Rapid on‐site determination of melamine in raw milk by an immunochromatographic strip

A rapid and simple method was established based on gold nanoparticle‐labelled monoclonal antibody probes for the detection of melamine pollution in raw milk. The conditions for conjugation between the antibody and gold nanoparticles were optimised (pH 8.0, antibody concentration 5 μg mL^?1). It gives a single proportional to melamine concentration with a performance time of only 3 min. A practical calibration curve was established with a reader system with limit of detection calculated as 4.47 and 8.34 μg L^?1 with naked eyes. Three structural analogues, atrazine, desethyl‐desisopropyl‐atrazine and cyromazine, were used to test the specificity of the immunochromatographic strip, and small colour changes on the strip test line were found even at the 500 ng mL^?1 spiked level. Fifty raw milk samples were detected with this strip method, and the resulting data coincided well with results from gas chromatography tandem mass spectrometry. The above‐mentioned results showed that this test strip can be used for melamine screening in the daily monitoring of milk.     

Computer image analysis: an additional tool for the identification of processed poultry and mammal protein containing bones

The aims of this study were (1) to evaluate the potential of image analysis measurements, in combination with the official analytical methods for the detection of constituents of animal origin in feedstuffs, to distinguish between poultry versus mammals; and (2) to identify possible markers that can be used in routine analysis. For this purpose, 14 mammal and seven poultry samples and a total of 1081 bone fragment lacunae were analysed by combining the microscopic methods with computer image analysis. The distribution of 30 different measured size and shape bone lacunae variables were studied both within and between the two zoological classes. In all cases a considerable overlap between classes meant that classification of individual lacunae was problematic, though a clear separation in the means did allow successful classification of samples on the basis of averages. The variables most useful for classification were those related to size, lacuna area for example. The approach shows considerable promise but will need further study using a larger number of samples with a wider range.     

Field measurements of aerosol particle dry deposition on tropical foliage at an urban site

This paper presents dry deposition of major ions on tropical foliage (leaves of Ashok (Polyalthia longifolia) and Cassia (Cassia siamea)) at St. John's, Agra, an urban site of tropical India on nonrainy, nondewy, and nonfoggy days. The deposition flux was higher on Cassia leaf than Ashok leaf probably due to a rougher surface as shown by scanning electron microscopy. Dry deposition of cations varies from 0.46 to 12.16 mg m(-2) day(-1) while anions vary from 0.04 to 3.24 mg m(-2) day(-1). The percentage contribution of alkaline components is greater than that of acidic components, indicating the alkaline nature of dry deposition. Two-way analysis of variance results reveal significant seasonal variation only for K+, SO4(2-), and F-; however, values varied season to season for Na+, Ca2+, Mg2+, Cl-, NO3-, and NH4+ also. The large seasonal variation in deposition flux may be due to meteorological conditions, diameter of particles, and variation in atmospheric level. SO42- and NO3- show significant correlation, indicating their origin from similar sources while significant correlation between Ca2+ and Mg2+ implies their origin from soil. Poor correlation between Ca2+ and SO4(2-), Ca2+ and NO3-, and Mg2+ and SO4(2-) indicates that in addition to soil other sources also contribute to dry deposition. Low dry deposition fluxes of SO2- and NO3- compared to Ca2+ and Mg2+ may be due to low mass medium diameters of SO4(2-) and NO3- and may be due to uptake through the stomatal pores abundant on leaf surfaces. Factor analysis was employed to identify the sources. F-, Cl, SO4(2-), NO3-, and K+ are grouped together in the first factor, indicating their probable contribution from combustion, Ca2+, Mg2+, and NH4+ are grouped in factor II, which may be attributed to road dust and soil, and factor III includes mainly Na+ and F-, probably contributed from brick-kiln industries. Atmospheric concentrations of F-, Cl-, NOs-, SO4(2-), Na+, K+, Ca2+, Mg2+, and NH4+ were found to be 0.38, 2.28, 1.31, 2.74, 0.44, 0.59, 1.21, 1.2, and 2.29 microg m(-3), respectively.     

Hydrogen isotopic enrichment: an indicator of biodegradation at a petroleum hydrocarbon contaminated field site

Compound-specific carbon and hydrogen isotope analysis was used to investigate biodegradation of benzene and ethylbenzene in contaminated groundwater at Dow Benelux BV industrial site. delta13C values for dissolved benzene and ethylbenzene in downgradient samples were enriched by up to 2+/-0.5 per thousand, in 13C, compared to the delta13C value of the source area samples. delta2H values for dissolved benzene and ethylbenzene in downgradient samples exhibited larger isotopic enrichments of up to 27+/-5 per thousand for benzene and up to 50+/-5 per thousand for ethylbenzene relative to the source area. The observed carbon and hydrogen isotopic fractionation in downgradient samples provides evidence of biodegradation of both benzene and ethylbenzene within the study area at Dow Benelux BV. The estimated extents of biodegradation of benzene derived from carbon and hydrogen isotopic compositions for each sample are in agreement, supporting the conclusion that biodegradation is the primary control on the observed differences in carbon and hydrogen isotope values. Combined carbon and hydrogen isotope analyses provides the ability to compare biodegradation in the field based on two different parameters, and hence provides a stronger basis for assessment of biodegradation of petroleum hydrocarbon contaminants.