DNA methylation of DKK-1 may correlate with pathological bone formation in ankylosing spondylitis.

OBJECTIVE: To investigate DNA methylation (DNAm) status of dickkopf-associated protein 1 (DKK-1) in ossified hip capsule synovium and serum among patients with ankylosing spondylitis (AS). METHODS: Western blot was applied to detect the level of DKK-1 protein expression in hip joint capsule tissues from four patients with AS as well as four patients with femoral neck fracture (FNF) caused by trauma as control. DKK-1 gene promoter methylation (GPM) was examined by methylation-specific polymerase chain reaction. Reverse transcription-polymerase chain reaction was performed to examine the messenger RNA (mRNA) levels of DKK-1, ß-catenin, and Wnt3a in both tissue and serum. The DNAm status of serum DKK-1 was measured among 36 patients with AS and syndesmophytes (AS + syndesmophytes group), 40 patients with AS but no syndesmophyte (AS group), and 42 healthy individuals (control group). Also, the serum levels of DKK-1 were measured by enzyme-linked immunosorbent assay. The modified New York criteria (mNYC) together with the modified Stoke Ankylosing Spondylitis Spinal Score (mSASSS) were adopted to examine the radiographic progression of AS. The receiver operating characteristic (ROC) curve was applied to investigate the diagnostic value of the methylation rate of DKK-1 with regard to radiographic progression. RESULTS: The expressions of DKK-1 protein and mRNA in hip joint capsule tissues of AS patients were significantly lower, while DKK-1 GPM rate, ß-catenin mRNA, and Wnt3a mRNA were markedly higher when compared with FNF group. For serum samples, the DKK-1 methylation rate was significantly higher in AS+ syndesmophytes group in contrast to AS group and healthy controls. Serum levels of DKK-1 protein and mRNA in AS with syndesmophytes group were markedly decreased, while ß-catenin mRNA and Wnt3a mRNA expressions were significantly increased than AS with no syndesmophyte group and the healthy control group. AS patients in Grade 4 showed a significantly higher serum DKK-1 GPM rate than those in Grade 3 based on mNYC. Serum DKK-1 GPM level was markedly and positively correlated with mSASSS. Serum levels of DKK-1 in AS+ syndesmophytes group were markedly lower compared with AS but no syndesmophyte group and healthy controls. ROC curve analysis indicated that serum DKK-1 methylation rate serves as a decent indicator for AS radiographic progression. CONCLUSION: DNAm of DKK-1 may correlate with pathological bone formation in AS, which may provide new strategies for the treatment of AS abnormal bone formation.

[Characteristics of Nitrogen Removal by a Heterotrophic Nitrification-Aerobic Denitrification Bacterium YL].

Traditional process of autotrophic nitrification-anaerobic denitrification usually has problems of long procedure and low efficiency. To overcome these problems, a heterotrophic nitrification-aerobic denitrification bacterium YL was isolated from a domesticated mature SBR reactor with efficient simultaneous nitrification and denitrification ability, and was identified as Pseudomonas aeruginosa YL. Meanwhile, the characteristics of the nitrogen removal of strain YL was investigated through single-factor experiments and an orthogonal experiment. The results showed that the preferred conditions were: succinate as the carbon source, C/N ratio of 10, pH of 7.0, temperature of 30°C, and the shaking speed of 160-200 r · min(-1), while the removal rate of ammonia oxidation was 5. 05 mg · (g · h)(-1), the transformation rate of TOC was 45.95 mg · (g · h)(-1), and the removal rates of nitrogen and TOC were 100% and 90.8%, respectively. Nitrite, nitrate and hydroxylamine could also be metabolized by strain YL, and the removal rates were 92.7%, 93.6% and 94.8%, respectively. The most important influencing factor on aerobic denitrification of strain YL was C/N ratio. Under the optimal conditions (C/N = 10, T = 30°C , r = 200 r · min(-1), pH = 7), the removal rates of nitrate and total nitrogen were 94.6% and 76.3%, respectively. Hence, strain YL could remove nitrogen by heterotrophic nitrification-aerobic denitrification independently, quickly, and effectively.

Nitrogen removal characteristics of a heterotrophic nitrifier Acinetobacter junii YB and its potential application for the treatment of high-strength nitrogenous wastewater.

Acinetobacter junii YB was found to exhibit efficient heterotrophic nitrification and aerobic denitrification ability, with the maximum ammonium, nitrite and nitrate removal rate of 8.82, 8.45 and 7.98 mg/L/h, respectively. Meanwhile, ammonium was found to be removed preferentially in the process of simultaneous nitrification and denitrification in mixed N-sources. The successful PCR amplification of hao, napA and nirS genes further provided additional evidence of heterotrophic nitrification and aerobic denitrification by strain YB. In addition, orthogonal test showed that dissolved oxygen was the most important determinant of nitrite removal, and the optimal conditions were C/N 15, pH 7.0, 37 °C and 200 rpm. Furthermore, stable nitrogen and organics removal were achieved by one-time dosing of enriched bacteria in a sequencing batch reactor. The inoculation of strain YB significantly improved the denitrification efficiency with minimal accumulation of nitrified products, which demonstrated high potential of the isolate for future practical applications.