Detection of soluble methane monooxygenase producing Methylosinus trichosporium OB3b by polymerase chain reaction

The soluble methane monooxygenase (sMMO) enzyme complex of methanotrophs cometabolizes haloaliphatic compounds such as trichloroethylene. Two 18-mer oligonucleotides as primary primers and a nested primer of the same length were selected to amplify specific DNA sequences of the sMMO gene cluster using polymerase chain reaction (PCR). Two DNA fragments of sizes 270 and 400 base pairs were obtained when purified DNA from the methanotroph Methylosinus trichosporium OB3b was used as template. The primers were specific for sMMO sequences of M. trichosporium, since none of the 13 bacterial isolates screened yielded the expected length of PCR-amplified DNA fragments. The detection limit of the PCR method was 5 x 10(2) cells of M. trichosporium. The sMMO sequences were successfully amplified in groundwater (containing native microbial population) when seeded with M. trichosporium, FP1 sense (5'-ATGTCCAGCGCTCATAAC-3'), RP1 antisense (5'-TCAGATGTCGGTCAGGGC-3'), FP2 sense nested (5'GCCATCATCGGTCAGGGC-3'), and FP2 sense nested (5'-GCCATCATCGAGGACATC-3').     

Oxidation of trichloroethylene, 1,1-dichloroethylene, and chloroform by toluene/o-xylene monooxygenase from Pseudomonas stutzeri OX1

Toluene/o-xylene monooxygenase (ToMO) from Pseudomonas stutzeri OX1, which oxidizes toluene and o-xylene, was examined for its ability to degrade the environmental pollutants trichloroethylene (TCE), 1, 1-dichloroethylene (1,1-DCE), cis-1,2-DCE, trans-1,2-DCE, chloroform, dichloromethane, phenol, 2,4-dichlorophenol, 2,4,5-trichlorophenol, 2,4,6-trichlorophenol, 2,3,5,6-tetrachlorophenol, and 2,3,4,5, 6-pentachlorophenol. Escherichia coli JM109 that expressed ToMO from genes on plasmid pBZ1260 under control of the lac promoter degraded TCE (3.3 microM), 1,1-DCE (1.25 microM), and chloroform (6.3 microM) at initial rates of 3.1, 3.6, and 1.6 nmol/(min x mg of protein), respectively. Stoichiometric amounts of chloride release were seen, indicating mineralization (2.6, 1.5, and 2.3 Cl- atoms per molecule of TCE, 1,1-DCE, and chloroform, respectively). Thus, the substrate range of ToMO is extended to include aliphatic chlorinated compounds.     

Some properties of a soluble methane mono-oxygenase from Methylococcus capsulatus strain Bath

Soluble extracts of Methylococcus capsulatus (Bath), obtained by centrifugation of crude extracts at 160000g for 1h, catalyse the NAD(P)H- and O2-dependent disappearance of bromomethane, and also the formation of methanol from methane. Soluble methane mono-oxygenase is not inhibited by chelating agents or by most electron-transport inhibitors, and is a multicomponent enzyme.     

Effects of hydrogen sulfide containing gas condensate on the hepatic microsomal monooxygenase system]

Chronic action of different xenobiotics which form a part of condensed hydrogen sulphide-containing gas causes substantial changes in the activity of xenobiotic metabolic enzymes, which is suggestive of the damaged hepatic microsomal monooxygenase system. As the rate of biotransformation becomes higher, there are increases in the activity of microsomal monooxygenases and in the generation of active oxygen forms and hydrogen peroxide, by impairing antiradical and antiperoxide mechanisms. The experimentally used concentration of substances as constituents of condensed gas corresponds to the maximum acceptable air concentration of in the working area of gas-refining plants, but the functional features of the body's detoxifying system--the hepatic monooxygenase system should be taken into account while developing preventive measures of occupational diseases.     

Oxidation of zinc sulfide in a fluidized bed

Kinetics of oxidation of ZnS particles in a batch-type fluidized bed were studied at temperatures between 800 and 910°C. A two-phase model was employed for the fluidized bed, and the partial pressure of oxygen and the gas-film mass transfer coefficient on the particle surface were separately evaluated in gas bubbles and in the emulsion phase. The calculated fractional reaction coincided well with the experimental results. The difference in O₂ partial pressure between gas bubbles and emulsion phase was found to be fairly large especially under the vigorous fluidizing condition. Furthermore, it was shown from the mathematical model that the reaction of ZnS particles in the gas bubbles is negligible because of the extremely low solid concentration and that the overall rate of reaction in the emulsion phase is virtually controlled by the rate of gas-film mass transfer at higher temperature. The resistance of interfacial reaction within the particle also becomes significant when the temperature is lowered. Y. Fukunaka and T. Monta are both former Graduate Students, Kyoto University, Kyoto, Japan.     

Oxidation kinetic studies of zinc sulfide in a fluidized bed reactor

Kinetic studies of the oxidation of zinc sulfide were carried out in a fluidized bed reactor over a temperature range of 740° to 1000°C with O-N gas mixtures of 20 to 40 pct O₂. A mathematical model was developed to describe the overall conversion of the solids. Application of the model to the experimental data indicated that the chemical reaction at the outer boundary of the unreacted sulfide core was the rate-limiting step for the process. The temperature dependence of the kinetic constant corresponded to an activation energy of 40,250 cal per mole. Oxygen starvation in the bed was not limiting in any of the experimental runs, but an increase in the inlet-oxygen mole fraction resulted in a substantial increase in reaction rate.     

Oxidation rates of liquid copper and liquid copper sulfide

The kinetics of oxidation of liquid copper and copper sulfide were determined when the rate was controlled by the diffusion of the oxidizing gas to the melt interface. The method incorporated a capillary tube sample holder and the flux rate was measured by a quartz spring and cathetometer. The sample of copper sulfide was suspended from the balance in a tube furnace through which was passed a mixture of oxygen and argon gas. The oxidation of molten copper sulfide was found to proceed by the formation of copper followed by the formation of copper oxide. From the results, the diffusion coefficients of SO₂ and O₂ in argon were calculated over a temperature range of 1150° to 1350°C. The data compare well with the empirical equations for the diffusion of these gases. At lower temperatures, a different oxidation behavior is observed due to solidification of the sample.     

Copper-dependent reciprocal transcriptional regulation of methane monooxygenase genes in Methylococcus capsulatus and Methylosinus trichosporium

PURPOSE: To better understand the dose and time dependence of radiation therapy (RT)-induced regional lung dysfunction as assessed by changes in regional lung perfusion. METHODS AND MATERIALS: Patients who were to receive RT for tumors in and around the thorax, wherein portions of healthy lung would be incidentally irradiated, were prospectively studied. Regional function was assessed pre- and post-RT with single photon emission computed tomography (SPECT) lung perfusion scans, obtained following the intravenous administration of approximately 4 mCi of technetium-99m macroaggregated albumin. Pre-RT computed tomography (CT) scans were used to calculate the three-dimensional (3D) dose distribution, reflecting tissue density inhomogeneity corrections. Each SPECT scan was correlated with the pre-RT CT scan, and the 3D dose distribution. Changes in regional lung perfusion were correlated with regional RT dose, at various time intervals following radiation. RESULTS: The data from 20 patients (7 breast cancer, 5 lymphoma, 1 esophagus, 1 sarcoma, and 6 lung cancer) have been analyzed. Patients with gross intrathoracic lung cancers causing obstruction of regional pulmonary arteries were not included. For most patients, there is a statistically significant dose-dependent reduction in regional blood flow at all time points following radiation. While a time dependence is suggested in the high dose range, the limited amount of data prevents meaningful statistical evaluation. CONCLUSIONS: Radiation therapy-induced regional lung dysfunction occurs in a dose-dependent manner and develops within 3-6 months following radiation. In contrast to classical "sigmoid" dose-response curves, described mainly for changes following whole lung irradiation, these data suggest a more gradual relationship between regional dysfunction and RT dose. Retraction of irradiated lung with secondary movement of unirradiated lung into the "3D-defined irradiated volume" may have introduced inaccuracies into this analysis. Additional studies are currently underway to assess this possibility and better refine this dose-response curve. Studies are underway to determine if changes in assessments of whole lung function, such as pulmonary function tests, can be predicted by summing the regional changes observed.     

Anaerobic versus aerobic degradation of dimethyl sulfide and methanethiol in anoxic freshwater sediments

Degradation of dimethyl sulfide and methanethiol in slurries prepared from sediments of minerotrophic peatland ditches were studied under various conditions. Maximal aerobic dimethyl sulfide-degrading capacities (4.95 nmol per ml of sediment slurry. h-1), measured in bottles shaken under an air atmosphere, were 10-fold higher than the maximal anaerobic degrading capacities determined from bottles shaken under N2 or H2 atmosphere (0.37 and 0. 32 nmol per ml of sediment slurry. h-1, respectively). Incubations under experimental conditions which mimic the in situ conditions (i. e., not shaken and with an air headspace), however, revealed that aerobic degradation of dimethyl sulfide and methanethiol in freshwater sediments is low due to oxygen limitation. Inhibition studies with bromoethanesulfonic acid and sodium tungstate demonstrated that the degradation of dimethyl sulfide and methanethiol in these incubations originated mainly from methanogenic activity. Prolonged incubation under a H2 atmosphere resulted in lower dimethyl sulfide degradation rates. Kinetic analysis of the data resulted in apparent Km values (6 to 8 microM) for aerobic dimethyl sulfide degradation which are comparable to those reported for Thiobacillus spp., Hyphomicrobium spp., and other methylotrophs. Apparent Km values determined for anaerobic degradation of dimethyl sulfide (3 to 8 microM) were of the same order of magnitude. The low apparent Km values obtained explain the low dimethyl sulfide and methanethiol concentrations in freshwater sediments that we reported previously. Our observations point to methanogenesis as the major mechanism of dimethyl sulfide and methanethiol consumption in freshwater sediments.     

Human corneal studies with a vitrification solution containing dimethyl sulfoxide, formamide, and 1,2-propanediol

We tested the tolerance of human corneas to a vitrification solution, modified VS41A, containing 3.1 M dimethyl sulfoxide, 3.1 M formamide, and 2.2 M 1,2-propanediol in a carrier solution consisting of the corneal storage medium CPTES with 2.5% w/v chondroitin sulfate. Seven human corneas were exposed for 10 min each to graded concentrations of the solution at 0 degree C, remaining in the full-strength solution for 10 min. The corneas had significantly more endothelial cell damage (P < 0.05) than seven mated control corneas, but it was minimal (4.3% cell loss). Attempts at vitrification and rewarming of three corneas exposed to the solution by this protocol, however, resulted in ice formation in the peripheral corneal stroma and severe endothelial damage. Presumably, equilibration with the cryoprotectant in the thicker periphery of the human cornea had not occurred. Ice did not form on the center of one cornea, and substantial numbers of central endothelial cells survived after vitrification in this case. Immersion of the human corneas for 25 min in each of the four graded solutions at 0 degree C was required for sufficient penetration of the cryoprotectant to allow total corneal vitrification and rewarming without ice formation. This prolonged exposure to modified VS41A caused unacceptable damage to the corneal endothelium, however. Successful vitrification of human corneas with this solution will require a safe method for obtaining corneal equilibration with the cryoprotectant.     

硫化钠还原-硫酸铈滴定法测定含锑金精矿中锑

以甲基橙作指示剂采用硫酸铈滴定法对含锑金精矿中锑进行测定时,若样品中铁含量较高,则溶液中Fe?的颜色会对终点颜色变化产生严重干扰,使终点判断较为困难。经过多次试验发现,采用亚甲基蓝-甲基橙作指示剂能够消除Fe?对滴定终点的干扰,使终点颜色变化敏锐。据此,实验采用硫酸和硫酸钾溶解样品,用硫化钠将Sb(V)还原为Sb?,在盐酸介质中,加热溶液温度至70~90℃,以亚甲基蓝-甲基橙为指示剂,建立了硫酸铈滴定法测定含锑金精矿中锑的方法。采用实验方法对含锑金精矿实际样品进行精密度和加标回收试验,测定结果的相对标准偏差(RSD,n=8)小于4%,锑的加标回收率在99%~102%之间。将实验方法应用于含锑金精矿实际样品中锑的测定,所得结果与国家标准方法 GB/T 15925—2010的测定值相吻合。     

Column bioleaching of a low grade nickel-bearing sulfide ore containing high magnesium as olivine,chlorite and antigorite

This work investigates the bioleaching of Jinchuan low grade nickel-bearing sulfide ore containing rather high levels of olivine, chlorite and antigorite (MgO 30–35%) present in the main gangue minerals using a mixed mesophiles which are composed of Acidithiobacillus ferrooxidans and Acidithiobacillus thiooxidans. It aims to test the technical feasibility to recover valuable metals from Jinchuan low-grade nickel-bearing sulfide ore by bioleaching process. The tolerance of the mixed bacteria to Mg²⁺ could be improved markedly from 10 g/L to 25 g/L after nearly 2 years adaptation. A nickel recovery of 91% and a cobalt recovery of 81% were achieved in 312 days column leaching process including 60 days acid pre-leaching stage and 252 days bioleaching stage.     

含金锑砷多金属硫化矿浮选分离试验研究

对含金锑砷多金属硫化矿矿石经混合浮选后，混合精矿用碳酸钠（Ｎａ＿２ＣＯ＿３）调节矿浆ｐＨ至１０左右，添加硫化钠（Ｎａ＿２Ｓ）作辉锑矿抑制剂，浮选砷（金），可获得含锑４９．４４％，砷０．４４％，回收率εＳｂ８０．８５％的锑精矿和含砷１０．９６％，金７９．９４ｇ／ｔ，回收率ε＿Ａｓ７７．９８％，ｅ＿Ａｕ６８．７９％的砷（金）精矿，试验表明，碳酸钠和硫化钠的使用比例以３：１最好。     

Determination of sulfide capacities of CaO-SiO2 slags containing CaF2 and B2O3 by an encapsulation method

The sulfide capacities of CaO-SiO₂ melts containing CaF₂ and B₂O₃ have been measured by simultaneous equilibration of samples within quartz capsules at 1503°C. In this method, the compositions of both the condensed phases and the static gas environment adjust to es-tablish a mutual equilibrium. Although the sulfurizing potential,P _{S 2}/P _{O 2}, is neither preset nor directly measured, the sulfide capacities,C _s, of all samples within the capsule are related by the following equation:C _S(j)=(pct S)j/(pct S)i · C_S(i)· Hence, the sulfide capacities of all samples may be calculated from the measurement of the final sulfur content of each sample and the inclusion of a “reference” slag sample for which C_S is known from previous measurements. In the present study, quartz capsules containing as many as fifty slag samples including several reference samples have been equilibrated. With considerable difficulty the encapsulation-equilibration method atl503°C has been developed into an acceptable procedure. The influences of CaF₂ and B₂O₃ on the sulfide capacities of several CaO-SiO₂ melts have been measured. The results may be summarized as follows: 1) for melts of fixed CaO/SiO₂ ratio, the addition of CaF₂ increases sulfide capacity and the addition of B₂O₃ decreases sulfide capacity, 2) the substitution of CaF₂ for CaO does not alter sulfide capacity significantly, and 3) the substitution of B₂O₃ for SiO₂ increases sulfide capacity slightly. These results have been obtained for melts with CaO/SiO₂ mass ratios ranging from 1.00 to 1.28 and with flux additions up to 10 wt pct.     

Inhibition of microsomal lipid peroxidation and monooxygenase activities by eugenol

Previously we reported that eugenol (4-allyl-2-methoxyphenol) inhibits non-enzymatic peroxidation in liver mitochondria (E. Nagababu and N. Lakshmaiah, 1992, Biochemical Pharmacology. 43, 2393-2400). In the present study, we examined the effect of eugenol on microsomal mixed function oxidase mediated peroxidation using Fe+3-ADP-NADPH, carbon tetrachloride (CCL4)-NADPH and cumene hydroperoxide (CumOOH) systems. In the presence of eugenol the formation of thiobarbituric acid reactive substances (TBARS) was decreased in all the systems (IC50 values: 14 microM for Fe+3-ADP-NADPH, 4.0 microM for CCl4-NADPH and 15 microM for CumOOH). Oxygen uptake was also inhibited to a similar extent with Fe+3-ADP-NADPH and CumOOH systems. A comparative evaluation with other antioxidants showed that in Fe+3-ADP-NADPH and CumOOH systems, the antioxidant efficacy was in the order: butylated hydroxytoluene (BHT) > eugenol > alpha-tocopherol, while in CCl4-NADPH system the order was alpha-tocopherol > BHT > eugenol. Time course of inhibition by eugenol indicated interference in initiation as well as propagation of peroxidation. Eugenol did not inhibit cytochrome P-450 reductase activity but it inhibited P-450 - linked monooxygenase activities such as aminopyrine-N-demethylase, N-nitrosodimethylamine demethylase, benzo(a)pyrene hydroxylase and ethoxyresorufin-O-deethylase to different extents. However, CumOOH supported monooxygenases (aminopyrine-N-demethylase and benzo(a)pyrene hydroxylase) required much higher concentrations of eugenol for inhibition. The concentration of eugenol required to inhibit monooxygenase activities was more than that required to inhibit peroxidation in all the systems. Eugenol elicited type 1 changes in the spectrum of microsomal cytochrome P-450. These results suggest that the inhibitory effect of eugenol on lipid peroxidation is predominantly due to its free radical quenching ability. Eugenol significantly protected against the degradation of cytochrome P-450 during lipid peroxidation with all the systems tested. These findings suggest that eugenol has the potential to be used as a therapeutic antioxidant. Further evaluation may throw more light on this aspect.     

钙处理对中硫含量钢中硫化物形态影响的试验研究

通过实验室真空感应炉冶炼铸锭然后锻造成棒材的方法,研究了不同Mn/S（质量比）对含硫非调质钢中硫化物形态的影响.研究结果表明：提高Mn/S有利于改善铸态和棒材中硫化物的分布均匀性;随着Mn/S的提高,棒材中长宽比<3形态的硫化物由5%左右提高至25%以上,Mn/S提高有利于硫化物的球化或纺锤化控制;实验条件下,Mn/S应控制在40左右.     

Direct oxidation of copper sulfide by a biofilm of Acidithiobacillus ferrooxidans

The direct oxidation of synthetic copper sulfide by Acidithiobacillus ferrooxidans was studied under iron-free conditions in two different types of biofilm reactors. It was shown that copper was oxidized biologically in the absence of intermediate oxidants such as ferric ions, but at rates much lower than these due to the indirect mechanism within the iron reduction–oxidation cycle. The effect of temperature on the rate of copper sulfide biooxidation was studied and the results show that the optimal temperature for the direct copper sulfide oxidation by A. ferrooxidans was close to 35 °C. The variation of acidity of the liquid medium between pH 1.5 and 3.5 did not affect the oxidation rate significantly.