Interaction between Illite and a Pseudomonas stutzeri-Heavy Oil Biodegradation Complex.

Illite is a widely distributed clay mineral with huge reserves in Earth's crust, but its effect on heavy oil biodegradation is rarely reported. This study made an investigation of the interactions between illite and a Pseudomonas stutzeri-heavy oil complex (PstHO). Results showed that, although illite exerted a negative effect on P. stutzeri degrading heavy oil by inhibiting the biodegradation of 64 saturated hydrocarbons (SHs) and 50 aromatic hydrocarbons (AHs), it selectively stimulated the biodegradation of 45 AHs with a specific structure, and its biogenic kaolinization at room temperature (35 °C) and pressure (1 atm) was observed in PstHO for the first time. The finding points out for the first time that, in PstHO, illite may change the quasi-sequential of AHs biodegradation of heavy oil, as well as its kaolinization without clay intermediate.

Multi-Temperatures Pyrolysis Gas Chromatography: A Rapid Method to Differentiate Microorganisms.

The identification of microorganisms using single-temperatures pyrolysis gas chromatography (ST-PyGC) has limitations, for example, the risk of missing characteristic peaks that are essential to the chemotaxonomic interpretation. In this paper, we proposed a new multi-temperature PyGC (MT-PyGC) method as an alternative to ST-PyGC, without sacrificing its speed and quality. Six bacteria (three Gram-positive and three Gram-negative), one micro-fungus and one archaeon, representing microorganisms from different domains, were analyzed by MT-PyGC. It is found that MT pyrograms cover a more complete range of characteristic peaks than ST. Coupling with thermogravimetric analysis, chemotaxonomic information extracted from pyrograms by MT-PyGC have the potential for the differentiation of microorganisms from environments including deep subterranean reservoirs and biomass conversion/biofuel production.

Regulation of LncRNA00067110 on Proliferation, Apoptosis and Melanins Production of B16-F10 Cells by Targeting Cabyr / 中国生物化学与分子生物学报

Long non-coding RNA (lncRNA) is a type of non-coding RNA with the more than 200 nucleotides. Several lncRNAs have been identified as the potential targets for cancer therapy. LncRNA00067110 is one of the differentially expressed genes in the transcriptome profiles of melanoma B16-F10 cells compared to normal mice melanocytes. To investigate whether lncRNA00067110 regulates the proliferation, apoptosis and melanogenesis of B16-F10 cells, the calcium-binding tyrosine phosphorylation regulated protein (Cabyr) target gene was predicted by LncTar and verified by dual luciferase activities. The regulating function of lncRNA00067110 was investigated by the analysis of transcriptome profiles and to detect the proliferation, apoptosis and melanin production of B16-F10 cells transfected by the overexpression plasmids of lncRNA00067110. The results showed that the relationship of lncRNA00067110 targeting Cabyr, the mRNA and protein levels of proliferation (MEK/ERK/MNK/CREB) and melanogenesis-related genes (TYR family and CREB) were significantly down-regulated, while the mRNA and protein levels of apoptosis-related genes (AKT and Bcl-2) were up-regulated in B16-F10 cells with lncRNA00067110 overexpression. The transcriptome profile of B16-F10 cells with lncRNA00067110 overexpression showed that 17 genes were differentially expressed, among which Cabyr was up-regulated. Furthermore, the effect of lncRNA00067110 on the phenotypes of cell proliferation and apoptosis were verified. The results suggested that lncRNA00067110 might be a novel target for the treatment of melanoma by targeting Cabyr, which regulate the expression of related genes to inhibit the proliferation and melanogenesis, as well as to induce the apoptosis of B16-F10 cells.

A rapid change in microbial communities of the shale gas drilling fluid from 3548 m depth to the above-ground storage tank.

Despite the growing body of studies on the various fracturing phrases, the research on the differences between subterranean and surface microorganisms at shale gas drilling sites is still limited. Generally, shale gas development and the production process are divided into drilling and fracturing. The distribution of microbial communities in the latter has been paid some attention, but a deficit remains in terms of our understanding of the microbial community in the former, especially for the phase of drilling flowback and drilling flowback surface. In this study, four drilling flowback fluids (DFFs) (H230-flowback drilling cuttings, H23G-flowback drilling mud, H240-flowback drilling sediment, and H21F-flowback drilling water) from the outlet of subterranean pipeline to the inlet of storage tank were successively collected from H2 shale gas field during its initial drilling in Sichuan, China. Natural mountain water (H10W) used as the injection water of H2 was also sampled. Illumina MiSeq 16S rRNA gene sequencing revealed a total of 8 phyla, 17 classes, 36 orders, 62 families, and 98 genera that were recovered from these samples with uneven distribution. The majority of the obtained sequences belonged to the phyla Proteobacteria (75.36%), Bacteroidetes (10.75%), and Firmicutes (5.64%), with significant differences found in DFFs and injection water. The richness of microorganisms gradually increased with the increasing flowback flowing distance (H230 < H23G < H240 < H21F < H10W), which was employed to reveal a rapid change in microbiota that was evident in samples along the flow path aboveground from a depth of 3548 m. The findings of this study could expand our understanding of the ecological role of microorganisms during the shale gas drilling phase. Furthermore, the study highlights the temporal-spatial trajectory of microbial communities from subterranean environments to the surface in a short period of 30 days.

Cupidesulfovibrio liaohensis gen. nov., sp. nov., a novel sulphate-reducing bacterium isolated from an oil reservoir and reclassification of Desulfovibrio oxamicus and Desulfovibrio termitidis as Cupidesulfovibrio oxamicus comb. nov. and Cupidesulfovibrio termitidis comb. nov.

A novel sulphate-reducing, Gram-stain-negative, anaerobic strain, isolate XJ01T, recovered from production fluid at the LiaoHe oilfield, PR China, was the subject of a polyphasic study. The isolate together with Desulfovibrio oxamicus NCIMB 9442T and Desulfovibrio termitidis DSM 5308T formed a distinct, well-supported clade in the Desulfovibrionaceae 16S rRNA gene tree. The taxonomic status of the clade was underscored by complementary phenotypic data. The three isolates comprising the clade formed distinct phyletic branches and were distinguished using a combination of physiological features and by low average nucleotide identity and digital DNA-DNA hybridization values. Consequently, it is proposed that isolate XJ01T represents a novel genus and species for which the name Cupidesulfovibrio liaohensis gen. nov., sp. nov. is proposed with the type strain XJ01T (=CGMCC 1.5227T=DSM 107637T). It is also proposed that D. oxamicus and D. termitidis be reclassified as Cupidesulfovibrio oxamicus comb. nov. and Cupidesulfovibrio termitidis comb. nov., respectively.

DKK3 Regulates Melanogenesis in Melanocytes of Alpaca / 中国生物化学与分子生物学报

Dickkopf-3 (DKK3) , as a critical inhibitor of the Wnt/p-catenin signaling pathway, may he involved in melanogenesis.In the current study, we investigated the effects of DKK3 on melanogenesis in melanocytes of alpaca.Overexpression of DKK3 in alpaca melanocytes, the expression of Wntl, Lefl , Myc and the major target genes termed microphthalmia-associated transcription factor (M1TF) and its downstream genes, including tyrosinase (TYR), tyrosinase-related protein 1 (TYRP1) and tyrosinase- related protein 2 (TYRP2) were significantly decreased at both mRNA and protein levels (P<0.05); total alkali melanin, pheomelanin and eumelanin were decreased by 80.30%, 72.17% and 64.60% (P <0.05), respectively.In contrast, in the melanocytes transfected with siRNA-DKK3 (a small interference RNA targeting DKK3) , the expression of Wntl, Lefl, Myc, MITF, TYR, TYRPl and TYRP2 were significantly increased at both mRNA and protein levels (P<0.05) ; total alkali melanin, pheomelanin and eumelanin were significantly increased by 1.65 folds, 1.25 folds and 1.21 folds (P< 0.05) , respectively.These results indicate that DKK3 regulates melanogenesis in alpaca melanocytes via the Wnt/p-catenin signaling pathway and down-regulates MITF.

Unitary and binary remediations by plant and microorganism on refining oil-contaminated soil.

Refining oil contaminants are complex and cause serious harm to the environment. Remediation of refining oil-contaminated soil is challenging but has significant impact in China. Two plant species Agropyron fragile (Roth) P. Candargy and Avena sativa L. and one bacterium Bacillus tequilensis ZJ01 were used to investigate their efficiency in remediating the refining oil-polluted soil sampled from an oil field in northern China. The simulated experiments of remediations by A. fragile or A. sativa alone and A. fragile or A. sativa combined with B. tequilensis ZJ01 for 39 days and by B. tequilensis ZJ01 alone for 7 days were performed in the laboratory, with B. tequilensis ZJ01 added before or after the germination of seeds. Seed germination rates and morphological characteristics of the plants, along with the varieties of oil hydrocarbons in the soil, were recorded to reflect the remediation efficiency. The results showed that the contamination was weakened in all experimental groups. A. sativa was more sensitive to the pollutants than A. fragile, and A. fragile was much more resistant to the oil hydrocarbons, especially to aromatic hydrocarbons. Adding B. tequilensis ZJ01 before the germination of seeds could restrain the plant growth while adding after the germination of A. fragile seeds notably improved the remediation efficiency. The degradation rate of oil hydrocarbons by B. tequilensis ZJ01 alone was also considerable. Together, our results suggest that the unitary remediation by B. tequilensis ZJ01 and the binary remediation by A. fragile combined with B. tequilensis ZJ01 added after the germination of seeds are recommended for future in situ remediations.

Effect of anti-androgen and progesterone on development of regrowth antlers in 6 year old sika deer / 中国中药杂志

To investigate the effects of anti-androgen drugs and melengestrol acetate (MGA) on development of regrowth antlers in 6 year old sika deer, twenty healthysika deerwith similar body weight and antler weightwere randomly divided into five groups by using single factor test design: flutamide (=4), bicalutamide (=4), progesterone acetate (CPA, =4), melengestrol acetate (MGA, =4), control(=4). All deer were fed with same diets and were housed outside together in an opened fence of 15 m×30 m with free access to water and feed. Treatment groups were injected subcutaneously sustained-release agents of the four drugs respectively when two-branched antlers were harvested. The control group had no special treatment. In the experiment period of 60 d, blood sampleswere collected for 4 times for each deer. The concentration of testosterone in plasma was tested and analyzed to compare the changes between different groups. Development of regrowth antlers was observed. At the end of the experiment, regrowth antlers were weighted and analyzed. The resultsshowed that the weights of regrowth antlers in treatment groups were significantly greater than those from control group and the weight gain (as compared with the control group) was 100.50%, 64.46%, 87.16% and 117.46% respectively in flutamide group, bicalutamide group, progesterone acetate group and melengestrol acetate group. For plasma testosterone concentration, it was not significantly different in the early stage (in the first 35 d), but at the end of the experimen, it was significantly higher than that of earlier stage (<0.01) in various groups. Testosterone concentration of flutamide treated group was significantly lower than that of the other groups (<0.01), while the level inbicalutamide and MGA treated groups was significantly higher than that in other groups (<0.01). The results showed that both anti-androgen drugs and MGA treatment promoted the development of regrowth antlers and increased the weight of regrowth antlers, where the effect was most significant by MGA treatment. From the morphological observation of the antlers, it was found that anti-androgen and MGA treatments prolonged the growth period of regrowth antlers through delaying the ossification of antlers. However, plasma testosterone concentration was not affected by the treatments.

Mechanism of monolignol biotransformation by Rhus laccases in water-miscible organic solutions.

Several important monolignols such as coniferyl alcohol were catalyzed using Rhus laccase (RL) from Rhus vernicifera in a water/acetone solution. The enzymatic mechanism is discussed in detail. Sites 6, ß, and phenolic oxygen were the main active sites of phenylpropanoid compounds, which were first oxidized by the enzyme and then radicalized. RL was also responsible for lignin biosynthesis, especially in the early stage.

Effects of organic solvents on the activity of free and immobilised laccase from Rhus vernicifera.

Rhus laccase (RL) was covalently immobilised onto chitosan, and the effects of immobilisation on pH optimum, enzyme activity, thermostability, and re-use evaluated, using either N,N-dimethyl-p-phenylenediamine or 2,6-dimethoxyphenol as substrate. Immobilisation greatly enhanced enzyme thermostability, resulted in negligible loss of activity, and showed excellent re-use potential, with >80% relative activity retained after 15 cycles in aqueous solvent. Immobilised Rhus laccase (I-RL) was more catalytically active in both hydrophobic and hydrophilic organic solvents than free RL. With water-immiscible organic solvents, both free RL and I-RL required a minimum water content to achieve activity. With water-miscible organic solvents, in general a water content of ∼20-50% (v/v) was required to achieve activity using free RL, whereas with I-RL less water was generally required to achieve enzyme activity, and therefore considerably higher relative activity was exhibited at lower water contents. Kinetic investigations showed that the rate of substrate disappearance generally followed a pseudo-first-order law, and for evaluated water-immiscible organic solvents rate constants generally increased with decrease of hydrophobicity, however, in water-miscible organic solvents no such relationship was observed. Some discussion of the potential interactions between organic solvent molecules and enzyme active centres was provided to explain obtained results.

Effects of lacquer polysaccharides, glycoproteins and isoenzymes on the activity of free and immobilised laccase from Rhus vernicifera.

The purified polysaccharides, glycoproteins, and isoenzymes of Rhus laccase, and crude enzymes, from Chinese lacquer (Rhus vernicifera sap) were used to determine their influence on the enzymic activity of Rhus laccase on several substrates (4-phenylenediamine, isoeugenol and coniferyl alcohol). No product identity changes were observed when these components were added singularly or in combination to the enzymic reactions (only relative product yields varied significantly), however, the polysaccharides (GP1 and GP2) and glycoprotein (stellacyanin, St) exhibited negative effects, and the two isoenzymes (L1 and L2) exhibited positive synergistic effects, on the activity of Rhus laccase. With respect to the activity of the crude enzymes, the negative effects of GP1, GP2 and St were greater than the positive effects of L1 and L2, compared with free Rhus laccase on its own (using 4-phenylenediamine as substrate), the estimated inhibitory effect (of GP1, GP2 and St) being by at least a factor of 50 (even with the positive effect of L1 and L2). This contributes to understanding of lacquer storage stability and drying rates. Immobilisation of crude enzymes using a variety of techniques (using natural and modified polysaccharides, and an inorganic support) where evaluated using isoeugenol as substrate. Agar embedding and zirconium chloride chelation methods resulted in the highest substrate conversion levels. The yields and products of isoeugenol catalysis using Vietnamese crude enzymes/purified Rhus laccase and commercial Denilite laccase were also compared and contrasted with their Chinese lacquer sap equivalents.

Does Donglan lacquer tree belong to Rhus vernicifera species?

The lacquer trees in Donglan of Guangxi Province, China, were identified totally as the species Rhus succedanea found in Vietnam and Taiwan region, based on the results of pyrolysis-gas chromatography/mass spectrometry (Py-GC/MS), an easy and effective method to identify species of trees among those with similar properties. Analyses by IR and NMR, the drying properties, and conventional morphology also confirmed that the Donglan lacquer trees do not belong to Rhus vernicifera, like most trees of the China mainland. Some differences, however, such as the enzymatic activity and the components of the lacquer, were found between the Donglan lacquer and the Vietnam lacquer. The Donglan lacquer has a shorter drying time than the latter.