Genome Sequencing Reveals the Potential of Achromobacter sp. HZ01 for Bioremediation.

Petroleum pollution is a severe environmental issue. Comprehensively revealing the genetic backgrounds of hydrocarbon-degrading microorganisms contributes to developing effective methods for bioremediation of crude oil-polluted environments. Marine bacterium Achromobacter sp. HZ01 is capable of degrading hydrocarbons and producing biosurfactants. In this study, the draft genome (5.5 Mbp) of strain HZ01 has been obtained by Illumina sequencing, containing 5,162 predicted genes. Genome annotation shows that "amino acid metabolism" is the most abundant metabolic pathway. Strain HZ01 is not capable of using some common carbohydrates as the sole carbon sources, which is due to that it contains few genes associated with carbohydrate transport and lacks some important enzymes related to glycometabolism. It contains abundant proteins directly related to petroleum hydrocarbon degradation. AlkB hydroxylase and its homologs were not identified. It harbors a complete enzyme system of terminal oxidation pathway for n-alkane degradation, which may be initiated by cytochrome P450. The enzymes involved in the catechol pathway are relatively complete for the degradation of aromatic compounds. This bacterium lacks several essential enzymes for methane oxidation, and Baeyer-Villiger monooxygenase involved in the subterminal oxidation pathway and cycloalkane degradation was not identified. These results suggest that strain HZ01 degrades n-alkanes via the terminal oxidation pathway, degrades aromatic compounds primarily via the catechol pathway and cannot perform methane oxidation or cycloalkane degradation. Additionally, strain HZ01 possesses abundant genes related to the metabolism of secondary metabolites, including some genes involved in biosurfactant (such as glycolipids and lipopeptides) synthesis. The genome analysis also reveals its genetic basis for nitrogen metabolism, antibiotic resistance, regulatory responses to environmental changes, cell motility, and material transport. The obtained genome data provide us with a better understanding of hydrocarbon-degrading bacteria, which may contribute to the future design of rational strategies for bioremediation of petroleum-polluted marine environments.

Molecular characterization and gene expression of cathepsin L in Nile tilapia (Oreochromis niloticus).

Cathepsin L (CatL) has been widely known for its involvement in the innate immunity. However, it still remains poorly understand how CatL modulates the immune system of teleosts. Moreover, the CatL of Nile tilapia (NtCatL) has not been cloned or characterized. In this study, the gene encoding NtCatL was cloned, and was characterized by bioinformatics analysis, heterologous expression and protease activity assay. The coding sequence of NtCatL is 1017 bp in length and encodes 338 amino acid residues with a predicted molecular weight of 38.487 kDa and a theoretical isoelectric point of 5.79. NtCatL possesses the features of a typical cathepsin L, including one signal peptide, one propeptide region, and one papain family cysteine protease domain containing four active site residues (Gln135, Cys141, His281, and Asn305). The prediction of protein-protein interaction shows that NtCatL may interact with some functional proteins for realizing an immune function. Real-time quantitative PCR revealed the widespread transcriptional expression of NtCatL in six tissues of healthy Nile tilapia, and the NtCatL mRNA is significantly up-regulated after Streptococcus agalactiae challenge. These results suggest that NtCatL is likely to be involved in the immune reaction of Nile tilapia. Recombinant proteins from the mature domain (residues 117-337) of NtCatL were obtained by heterologous expression using pET28a and Rosetta (DE3) competent cells. A protein product with the high purity was obtained by using TALON Superflow purification rather than adopting HisTrap HP columns. The protease activity of the recombinant protein was verified by using a substrate hydrolyzing assay. This work has cloned and characterized the CatL from Nile tilapia for the first time, and contributes to elucidating the immunological functions of CatL.

Isolation and characterization of Burkholderia fungorum Gan-35 with the outstanding ammonia nitrogen-degrading ability from the tailings of rare-earth-element mines in southern Jiangxi, China.

The exploitation of rare-earth-element (REE) mines has resulted in severe ammonia nitrogen pollution and induced hazards to environments and human health. Screening microorganisms with the ammonia nitrogen-degrading ability provides a basis for bioremediation of ammonia nitrogen-polluted environments. In this study, a bacterium with the outstanding ammonia nitrogen-degrading capability was isolated from the tailings of REE mines in southern Jiangxi Province, China. This strain was identified as Burkholderia fungorum Gan-35 according to phenotypic and phylogenetic analyses. The optimal conditions for ammonia-nitrogen degradation by strain Gan-35 were determined as follows: pH value, 7.5; inoculum dose, 10%; incubation time, 44 h; temperature, 30 °C; and C/N ratio, 15:1. Strain Gan-35 degraded 68.6% of ammonia nitrogen under the optimized conditions. Nepeta cataria grew obviously better in the ammonia nitrogen-polluted soil with strain Gan-35 than that without inoculation, and the decrease in ammonia-nitrogen contents of the former was also more obvious than the latter. Besides, strain Gan-35 exhibited the tolerance to high salinities. In summary, strain Gan-35 harbors the ability of both ammonia-nitrogen degradation at high concentrations and promoting plant growth. This work has reported a Burkholderia strain with the ammonia nitrogen-degrading capability for the first time and is also the first study on the isolation of a bacterium with the ammonia nitrogen-degrading ability from the tailings of REE mines. The results are useful for developing an effective method for microbial remediation of the ammonia nitrogen-polluted tailings of REE mines.

[Manipulative reduction and percutaneous pin fixation for the treatment of severely displaced distal tibial fractures in children].

OBJECTIVE: To study the therapeutic efficacy of manipulative reduction and percutaneous pin fixation for the treatment of severely displaced distal tibial fractures in children. METHODS: From May 2009 to December 2012,56 children with severely displaced distal tibial fractures were analyzed, who had been treated with manipulative reduction and percutaneous pin fixation. Preoperative fractures were confirmed as severely displaced fractures by X-ray apparatus. There were 33 boys and 23 girls,ranging in age from 3 to 14 years, with an average of 10.1 years. All the fractures were closed without vascular or nerve injuries. According to the bone fracture type, under C-arm fluoroscopy, functional reduction was achieved by manipulative reduction. The fractures were fixed with percutaneous pins. Postoperative X-ray confirmed the functional reduction. Follow-up indexes were recorded: intra-operative and postoperative complications,postoperative radiographic examination, lower extremity length and range of ankle motion. Ankle score system of Teeny was used to evaluate ankle function. RESULTS: All the patients were followed up, and the duration ranged from 3 to 46 months, with an average duration of 19.4 months. According to the Teeny score standard, 35 patients got an excellent result, 7 good and 3 fair. Pin track reaction was found in 4 cases. The X-ray showed all the fractures healed without shortening deformity or epiphyseal arrest at early stage. All the patients could participate in the normal physical activities, having a normal range of motion and excellent strength of the ankle joint. CONCLUSION: The method of manipulative reduction and percutaneous pin fixation is a safe and convenient treatment for severely displaced distal tibial fractures in children. It has several advantages as follow: micro-trauma, tiny tissue damage, firm fixation, and the patients can exercise the function of ankle early, suggesting that it is an effective treatment method.

[Treatment type C fracture of the distal radius with locking compression plate and external fixators].

OBJECTIVE: To compare efficacy of unilateral external fixators and locking compression plates in treating type C fractures of the distal radius. METHODS: From January 2009 to June 2010, 76 patients with distal radius fracture were treated with LCP and external fixators, 54 patients were followed up. Among them, 29 cases were male and 25 cases were female with an average age of 45.31 (ranged, 24 to 68) years old. There were 29 patients in LCP group. According to AO classification, 8 cases were type C1, 7 cases were type C2 and 14 cases were type C3. There were 25 cases in external fixators group. According to AO classification, 6 cases were type C1, 8 cases were type C2 and 11 cases were type C3. Radial height, volar tilt and radial inclination were compared, advanced Gartland-Werley scoring were used to assessed wrist joint function after 6 and 12 months' following up. RESULTS: Two cases were suffered from nail infection in external fixators group. Fifty-four patients were followed up from 12 to 24 months with an average of 21.3 months. Radial height was (9.60 +/- 0.72) mm, volar tilt was (9.55 +/- 0.80) degrees and radial inclination was (21.40 +/- 0.78) degrees in LCP group,while those were (9.40 +/- 0.70) mm, (9.47 +/- 0.71) degrees and (21.20 +/- 0.73) degrees in external fixtors group, and with no statistical significance (P>0.05). Advanced Gartland-Werley score after 6 months' following up was 3.31 +/- 1.17 in LCP group, 5.56 +/- 1.58 in external fixtors group, and with significant difference (t=-5.99,P<0.05); after 12 months' following up, advanced Gartland-Werley score was respectively 2.66 +/- 1.01 and 3.08 +/- 1.00, but with no statistical meaning (t=-1.55, P>0.05). CONCLUSION: LCP and external fixtors can receive good curative effects in treating type C distal radius fracture, and LCP can obtain obviously short-term efficacy, while there is no significant difference between two groups in long-term results. For serious distal radius comminuted fracture which unable to plate internal fixation, external fixators is a better choice.