Functional metagenomic analysis of quorum sensing signaling in a nitrifying community.

Quorum sensing (QS) can function to shape the microbial community interactions, composition, and function. In wastewater treatment systems, acylated homoserine lactone (AHL)-based QS has been correlated with the conversion of floccular biomass into microbial granules, as well as EPS production and the nitrogen removal process. However, the role of QS in such complex communities is still not fully understood, including the QS-proficient taxa and the functional QS genes involved. To address these questions, we performed a metagenomic screen for AHL genes in an activated sludge microbial community from the Ulu Pandan wastewater treatment plant (WWTP) in Singapore followed by functional validation of luxI activity using AHL biosensors and LC-MSMS profiling. We identified 13 luxI and 30 luxR homologs from the activated sludge metagenome. Of those genes, two represented a cognate pair of luxIR genes belonging to a Nitrospira spp. and those genes were demonstrated to be functionally active. The LuxI homolog synthesized AHLs that were consistent with the dominant AHLs in the activated sludge system. Furthermore, the LuxR homolog was shown to bind to and induce expression of the luxI promoter, suggesting this represents an autoinduction feedback system, characteristic of QS circuits. Additionally, a second, active promoter was upstream of a gene encoding a protein with a GGDEF/EAL domain, commonly associated with modulating the intracellular concentration of the secondary messenger, c-di-GMP. Thus, the metagenomic approach used here was demonstrated to effectively identify functional QS genes and suggests that Nitrospira spp. maybe QS is active in the activated sludge community.

Long-term PM2.5 exposure increases the risk of non-small cell lung cancer (NSCLC) progression by enhancing interleukin-17a (IL-17a)-regulated proliferation and metastasis.

PM2.5 is a class of airborne particles and droplets with sustained high levels in many developing countries. Epidemiological studies have indicated that PM2.5 is closely associated with the increased morbidity and mortality of lung cancer in the world. Unfortunately, the effects of PM2.5 on lung cancer are largely unknown. In the present study, we attempted to explore the role of PM2.5 in the etiology of NSCLC. Here, we found that long-term PM2.5 exposure led to significant pulmonary injury. Epithelial-mesenchymal transition (EMT) and cancer stem cells (CSC) properties were highly induced by PM2.5 exposure. EMT was evidenced by the significant up-regulation of MMP2, MMP9, TGF-ß1, α-SMA, Fibronectin and Vimentin. Lung cancer progression was associated with the increased expression of Kras, c-Myc, breast cancer resistance protein BCRP (ABCG2), OCT4, SOX2 and Aldh1a1, but the decreased expression of p53 and PTEN. Importantly, mice with IL-17a knockout (IL-17a-/-) showed significantly alleviated lung injury and CSC properties following PM2.5 exposure. Also, IL-17a-/--attenuated tumor growth was recovered in PM2.5-exposed mice injected with recombinant mouse IL-17a, accompanied with significantly restored lung metastasis. Taken together, these data revealed that PM2.5 could promote the progression of lung cancer by enhancing the proliferation and metastasis through IL-17a signaling.

Ridge expansion alone or in combination with guided bone regeneration to facilitate implant placement in narrow alveolar ridges: a retrospective study.

OBJECTIVE: To evaluate the long-term outcomes of ridge expansion technique in dealing with horizontal bony insufficiency of alveolar ridges for implant placement. MATERIALS AND METHODS: During the period 2004-2009, 168 patients with width insufficiency of alveolar ridges were treated using the ridge expansion technique to obtain an improved bony base for implant placement. Depending on the severity of width insufficiency, the surgical procedures were classified into two groups: ridge expansion alone (Group 1) and ridge expansion in combination with guided bone regeneration (Group 2). After 4-6 months of unloaded healing, the implants were restored. The patients were followed up until 2013 with clinical and radiographic examinations. RESULTS: Among the 168 patients, 11 patients underwent a fracture of labial/buccal bony plate during surgery, which was corrected by changing the procedure into bone grafting, yielding a surgical failure rate of 6.5%. In the remaining 157 patients successfully treated by ridge expansion alone or in combination with GBR, 226 implants were simultaneously placed as planned. No implant failed over 2.8 years (6 months to 8 years) of follow-up, yielding a cumulative implant survival rate of 100% in each group. Six implants in Group 1 and 4 implants in Group 2, although osseointegrated and in function, did not fulfill success criteria: Cumulative implants' success rates were 93.2% in Group 1 and 95.6% in Group 2. The mean marginal bone losses during the first year in Group 1 and Group 2 were 0.69 and 0.43 mm, respectively, followed by an annual loss of ~ 0.06 and 0.07 mm, respectively, in the following years. No clinical parameter was abnormal. Twenty-two (10.4%) implants were exposed to peri-implant mucositis, whereas 19 (11.0%) implant-supported restorations were involved in prosthetic complications. CONCLUSIONS: The preliminary results of this retrospective study indicate that ridge expansion alone or in combination with GBR can be considered an effective and safe procedure for treatment of width insufficiency of alveolar ridges on the purpose of implant application.

Ischemic preconditioning enhances autophagy but suppresses autophagic cell death in rat spinal neurons following ischemia-reperfusion.

Autophagy serves to eliminate damaged proteins and organelles under normal physiological conditions and can be accelerated by pathological stress, possibly as a cytoprotective mechanism. Brief periods of ischemia (ischemic preconditioning or IPC) can reduce neuronal death in response to subsequent severe ischemic insults. Ischemic preconditioning also induces autophagy, but the contribution of autophagy to IPC-associated neuroprotection remains unclear. We investigated the contribution of autophagy to IPC-mediated neuroprotection in rats subjected to ischemic spinal cord injury. Fifty adult rats were randomly assigned to either (1) a sham group receiving anesthesia and surgical preparation (n=5), (2) an ischemia/reperfusion (I/R) group (n=20) subjected to 0.5 h ischemia followed by 3, 6, 12, or 24 h reperfusion, (3) an IPC group receiving three cycles of 5 min ischemia followed by 5 min of reperfusion (n=5), or (4) an IPC+I/R group (n=20). Hematoxylin-eosin (HE) and immunohistochemical staining were performed to evaluate spinal neuron survival in the four treatment groups. Autophagic activity was investigated by electron microscopy and by immunohistochemical and Western blot analyses of the autophagosome marker LC3-II and the autophagy-associated BH3 protein Beclin-1. Changes in Bcl-2/Beclin-1 complex association and Bcl-2 phosphorylation (p-Bcl-2) were examined by co-immunoprecipitation and Western blot analyses. In the I/R group, LC3-II was significantly elevated after 3h of reperfusion, but declined significantly by 24 h. At 24 h, I/R rats exhibited extensive spinal damage and decreased neuronal survival. In the IPC+IR group, neuronal death was reduced and expression of LC3-II sustained throughout the 24 h reperfusion period. In the I/R group, expression of (inactive) p-Bcl-2(Ser70) was increased significantly during reperfusion and was accompanied by dissociation of the Bcl-2/Beclin-1 complex and increased Beclin-1 expression. Preconditioning inhibited these changes in p-Bcl-2, Beclin-1, and Bcl-2/Beclin-1 complex expression. Ischemic preconditioning appears to sustain the beneficial effects of autophagic lysosomal degradation during I/R while inhibiting autophagic cell death.

Separation and determination of trace environmental estrogen through molecularly imprinted solid phase extraction coupled to high performance liquid chromatography.

In recent years, there has been an increasing demand on the determination of trace environmental estrogen. In this study, a new 17ß-estradiol molecularly imprinted polymer was successfully synthesized on the surface of activated silica gel. This prepared molecularly imprinted polymer was characterized, and found to exhibit good recognition, high selectivity, and a rapid adsorption-desorption dynamic toward 17ß-estradiol. Using the molecularly imprinted polymer as a sorbent, an effective method of solid phase extraction coupled to high performance liquid chromatography for the separation and determination of 17ß-estradiol was developed. Under optimal conditions, the detection limit (S/N = 3) of 17ß-estradiol was 12.0 ng L(-1). The enrichment factor was 198, and the linear range of the calibration graph was 0.05-300 mg L(-1). Blank water samples spiked with 17ß-estradiol at three different levels were analyzed by the developed method to afford recoveries ranging from 82.7% to 103.0%. Moreover, this presented method was applied to the quantitative detection of 17ß-estradiol in a polluted water sample with a level of 0.024 mg L(-1).