Strong succession in prokaryotic association networks and community assembly mechanisms in an acid mine drainage-impacted riverine ecosystem.

Acid mine drainage (AMD) serves as an ideal model system for investigating microbial ecology, interaction, and assembly mechanism in natural environments. While previous studies have explored the structure and function of microbial communities in AMD, the succession patterns of microbial association networks and underlying assembly mechanisms during natural attenuation processes remain elusive. Here, we investigated prokaryotic microbial diversity and community assembly along an AMD-impacted river, from the extremely acidic, heavily polluted headwaters to the nearly neutral downstream sites. Microbial diversity was increased along the river, and microbial community composition shifted from acidophile-dominated to freshwater taxa-dominated communities. The complexity and relative modularity of the microbial networks were also increased, indicating greater network stability during succession. Deterministic processes, including abiotic selection of pH and high contents of sulfur and iron, governed community assembly in the headwaters. Although the stochasticity ratio was increased downstream, manganese content, microbial negative cohesion, and relative modularity played important roles in shaping microbial community structure. Overall, this study provides valuable insights into the ecological processes that govern microbial community succession in AMD-impacted riverine ecosystems. These findings have important implications for in-situ remediation of AMD contamination.

Long-term elevated precipitation induces grassland soil carbon loss via microbe-plant-soil interplay.

Global climate models predict that the frequency and intensity of precipitation events will increase in many regions across the world. However, the biosphere-climate feedback to elevated precipitation (eP) remains elusive. Here, we report a study on one of the longest field experiments assessing the effects of eP, alone or in combination with other climate change drivers such as elevated CO2 (eCO2 ), warming and nitrogen deposition. Soil total carbon (C) decreased after a decade of eP treatment, while plant root production decreased after 2 years. To explain this asynchrony, we found that the relative abundances of fungal genes associated with chitin and protein degradation increased and were positively correlated with bacteriophage genes, suggesting a potential viral shunt in C degradation. In addition, eP increased the relative abundances of microbial stress tolerance genes, which are essential for coping with environmental stressors. Microbial responses to eP were phylogenetically conserved. The effects of eP on soil total C, root production, and microbes were interactively affected by eCO2 . Collectively, we demonstrate that long-term eP induces soil C loss, owing to changes in microbial community composition, functional traits, root production, and soil moisture. Our study unveils an important, previously unknown biosphere-climate feedback in Mediterranean-type water-limited ecosystems, namely how eP induces soil C loss via microbe-plant-soil interplay.

Total flavonoids of hawthorn leaves promote motor function recovery via inhibition of apoptosis after spinal cord injury.

Flavonoids have been reported to have therapeutic potential for spinal cord injury. Hawthorn leaves have abundant content and species of total flavonoids, and studies of the effects of the total flavonoids of hawthorn leaves on spinal cord injury have not been published in or outside China. Therefore, Sprague-Dawley rats were used to establish a spinal cord injury model by Allen's method. Rats were intraperitoneally injected with 0.2 mL of different concentrations of total flavonoids of hawthorn leaves (5, 10, and 20 mg/kg) after spinal cord injury. Injections were administered once every 6 hours, three times a day, for 14 days. After treatment with various concentrations of total flavonoids of hawthorn leaves, the Basso, Beattie, and Bresnahan scores and histological staining indicated decreases in the lesion cavity and number of apoptotic cells of the injured spinal cord tissue; the morphological arrangement of the myelin sheath and nerve cells tended to be regular; and the Nissl bodies in neurons increased. The Basso, Beattie, and Bresnahan scores of treated spinal cord injury rats were increased. Western blot assays showed that the expression levels of pro-apoptotic Bax and cleaved caspase-3 were decreased, but the expression level of the anti-apoptotic Bcl-2 protein was increased. The improvement of the above physiological indicators showed a dose-dependent relationship with the concentration of total flavonoids of hawthorn leaves. The above findings confirm that total flavonoids of hawthorn leaves can reduce apoptosis and exert neuroprotective effects to promote the recovery of the motor function of rats with spinal cord injury. This study was approved by the Ethics Committee of the Guangxi Medical University of China (approval No. 201810042) in October 2018.

Deficiency of PDK1 in osteoclasts delays fracture healing and repair.

Bone fractures are common traumatic injuries of the musculoskeletal system. However, delayed union and non­union fractures are a major clinical problem that present significant socioeconomic burden to patients and the public health sector. The bone­resorbing osteoclasts and bone­forming osteoblasts serve important roles in the fracture repair/healing process. Osteoclast deficiency or decreased osteoblast activity negatively impacts fracture healing. We previously demonstrated that the specific deletion of the serine/threonine kinase 3­phosphoinositide­dependent protein kinase 1 (PDK1) in osteoclasts leads to abrogated osteoclast formation and bone resorption in response to receptor activator of nuclear factor­κB in vitro and protected mice against ovariectomized­induced bone loss and lipopolysaccharide­induced osteolysis in vivo. Given the importance of osteoclasts in fracture repair, we hypothesized that the specific loss of PDK1 in osteoclasts will alter the fracture healing process. Mice of tibial fracture were constructed, and tibial specimens were sampled at 7­, 14­, 21­ and 28­days post­fracture to observe the effect of PDK1 gene regulated osteoclasts on fracture healing process by X­ray radiography, microcomputed tomography scanning, histomorphological staining and biomechanical testing. The present study revealed, using the tibial fracture model, that the specific deletion of the PDK1 gene in osteoclasts impeded the fracture healing process by delaying the resorption of the cartilaginous callus and subsequent remodeling of immature woven bone to structurally and mechanically ensure lamellar bone is stronger. No effect on osteoblast bone formation and osteogenesis was observed, thus indicating that delayed fracture healing is primarily due to defective osteoclast activity. These results provide important clinical implications for the use of anti­resorptive agents, such as bisphosphonates, for the treatment of osteolytic conditions. Such anti­resorptive therapies may detrimentally delay fracture healing and repair.

PDK1 is important lipid kinase for RANKL-induced osteoclast formation and function via the regulation of the Akt-GSK3ß-NFATc1 signaling cascade.

Perturbations in the balanced process of osteoblast-mediated bone formation and osteoclast-mediated bone resorption leading to excessive osteoclast formation and/or activity is the cause of many pathological bone conditions such as osteoporosis. The osteoclast is the only cell in the body capable of resorbing and degrading the mineralized bone matrix. Osteoclast formation from monocytic precursors is governed by the actions of two key cytokines macrophage-colony-stimulating factor and receptor activator of nuclear factor-κB ligand (RANKL). Binding of RANKL binding to receptor RANK initiates a series of downstream signaling responses leading to monocytic cell differentiation and fusion, and subsequent mature osteoclast bone resorption and survival. The phosphoinositide-3-kinase (PI3K)-protein kinase B (Akt) signaling cascade is one such pathway activated in response to RANKL. The 3-phosphoinositide-dependent protein kinase 1 (PDK1), is considered the master upstream lipid kinase of the PI3K-Akt cascade. PDK1 functions to phosphorylate and partially activate Akt, triggering the activation of downstream effectors. However, the role of PDK1 in osteoclasts has yet to be clearly defined. In this study, we specifically deleted the PDK1 gene in osteoclasts using the cathepsin-K promoter driven Cre-LoxP system. We found that the specific genetic ablation of PDK1 in osteoclasts leads to an osteoclast-poor osteopetrotic phenotype in mice. In vitro cellular assays further confirmed the impairment of osteoclast formation in response to RANKL by PDK1-deficient bone marrow macrophage (BMM) precursor cells. PDK1-deficient BMMs exhibited reduced ability to reorganize actin cytoskeleton to form a podosomal actin belt as a result of diminished capacity to fuse into giant multinucleated osteoclasts. Notably, biochemical analyses showed that PDK1 deficiency attenuated the phosphorylation of Akt and downstream effector GSK3ß, and reduced induction of NFATc1. GSK3ß is a reported negative regulator of NFATc1. GSK3ß activity is inhibited by Akt-dependent phosphorylation. Thus, our data provide clear genetic and mechanistic insights into the important role for PDK1 in osteoclasts.

miR-136-5p Regulates the Inflammatory Response by Targeting the IKKß/NF-κB/A20 Pathway After Spinal Cord Injury.

BACKGROUND/AIMS: miR-136-5p participates in recovery after spinal cord injury (SCI) via an unknown mechanism. We investigated the mechanism underlying the involvement of miR-136-5p in the inflammatory response in a rat model of SCI. METHODS: Sprague-Dawley rat astrocytes were cultured in vitro to construct a reporter plasmid. Luciferase assays were used to detect the ability of miR-136-5p to target the IKKß and A20 genes. Next, recombinant lentiviral vectors were constructed, which either overexpressed miR-136-5p or inhibited its expression. The influence of miR-136-5p overexpression and miR-136-5p silencing on inflammation was observed in vivo in an SCI rat model. The expression of IL-1ß, IL-6, TNF-α, IFN-α, and related proteins (A20, IKKß, and NF-κB) was detected. RESULTS: In vitro studies showed that luciferase activity was significantly activated in the presence of the 3' untranslated region (UTR) region of the IKKß gene after stimulation of cells with miR-136-5p. However, luciferase activity was significantly inhibited in the presence of the 3'UTR region of the A20 gene. Thus, miR-136-5p may act directly on the 3'UTR regions of the IKKß and A20 genes to regulate their expression. miR-136-5p overexpression promoted the production of related cytokines and NF-κB in SCI rats and inhibited the expression of A20 protein. CONCLUSION: Overexpression of miR-136-5p promotes the generation of IL-1ß, IL-6, TNF-α, IFN-α, IKKß, and NF-κB in SCI rats but inhibits the expression of A20. Under these conditions, inflammatory cell infiltration into the rat spinal cord increases and injury is significantly aggravated. Silencing of miR-136-5p significantly reduces the protein expression results described after miR-136-5p overexpression and ameliorates the inflammatory cell infiltration and damage to the spinal cord. Therefore, miR-136-5p might be a new target for the treatment of SCI.

Fluoride Removal from Brackish Groundwaters by Constant Current Capacitive Deionization (CDI).

Charging capacitive deionization (CDI) at constant voltage (CV) produces an effluent stream in which ion concentrations vary with time. Compared to CV, charging CDI at constant current (CC) has several advantages, particularly a stable and adjustable effluent ion concentration. In this work, the feasibility of removing fluoride from brackish groundwaters by single-pass constant-current (SPCC) CDI in both zero-volt and reverse-current desorption modes was investigated and a model developed to describe the selective electrosorption of fluoride and chloride. It was found that chloride is preferentially removed from the bulk solution during charging. Both experimental and theoretical results are presented showing effects of operating parameters, including adsorption/desorption current, pump flow rate and fluoride/chloride feed concentrations, on the effluent fluoride concentration, average fluoride adsorption rate and water recovery. Effects of design parameters are also discussed using the validated model. Finally, we describe a possible CDI assembly in which, under appropriate conditions, fluoride water quality targets can be met. The model developed here adequately describes the experimental results obtained and shows how change in the selected system design and operating conditions may impact treated water quality.

Investigation of fluoride removal from low-salinity groundwater by single-pass constant-voltage capacitive deionization.

Capacitive deionization (CDI) is attracting increasing attention as an emerging technology for the facile removal of ionic species from water. In this work, the feasibility of fluoride removal from low-salinity groundwaters by single-pass constant-voltage CDI was investigated and a model developed to describe the dynamic fluoride electrosorption behavior. Effects of operating parameters including charging voltage and pump flow rate as well as impact of fluoride and chloride feed concentrations on the effluent fluoride concentration and equilibrium fluoride adsorption capacity were studied and the obtained data used to validate the model. Using the validated model, the effects of various design parameters, including arrangement of multiple CDI cells, on fluoride removal were assessed. Single-pass constant-voltage CDI was found to be effective in removing fluoride from low-salinity groundwaters but, as expected, removal efficiency was compromised in waters of high salinity. The relatively simple electrosorption model developed here provided a satisfactory description of both fluoride removal and current evolution and would appear to be a useful tool for prediction of CDI performance over a range of operating conditions, cell arrangements and feed water compositions though scope for model improvement exists.

Floc properties of polyaluminum ferric chloride in water treatment: The effect of Al/Fe molar ratio and basicity.

Producing flocs with desired properties is significant for contaminants removal in water treatment works. In this study, an inorganic composite coagulant, polyaluminum ferric chloride (PAFC), was prepared and used in surface water treatment, and the influence of Al/Fe molar ratio and basicity (B) on floc properties was investigated. The contribution of metal species analysis showed a competition relationship between Al and Fe in the pre-hydrolysis, while the monomeric contents decreased with the increase of B value. The investigation of floc properties was conducted on a laser scattering instrument, in terms of floc size, strength, recovery capacity and fractal dimension (Df). The largest floc size and the highest growth rate was achieved when Al/Fe=7:1 and B=1.5. Floc formed at the Al/Fe ratios of 5:1 and 7:1 were considered to be more compact. Meanwhile, the Df value increased when B value was increased. At Al/Fe=7:1 and B=1.5, strongest flocs were obtained. During the breakage period, the Df value increased. As lower shear was replaced, the floc size decreased continuously, with a further increase of Df value. However, after breakage at higher shear, all of the PAFC flocs showed capacity for regrowth and loose structures were formed.

The impact of pH on floc structure characteristic of polyferric chloride in a low DOC and high alkalinity surface water treatment.

The adjustment of pH is an important way to enhance removal efficiency in coagulation units, and in this process, the floc size, strength and structure can be changed, influencing the subsequent solid/liquid separation effect. In this study, an inorganic polymer coagulant, polyferric chloride (PFC) was used in a low dissolved organic carbon (DOC) and high alkalinity surface water treatment. The influence of coagulation pH on removal efficiency, floc growth, strength, re-growth capability and fractal dimension was examined. The optimum dosage was predetermined as 0.150 mmol/L, and excellent particle and organic matter removal appeared in the pH range of 5.50-5.75. The structure characteristics of flocs formed under four pH conditions were investigated through the analysis of floc size, effect of shear and particle scattering properties by a laser scattering instrument. The results indicated that flocs formed at neutral pH condition gave the largest floc size and the highest growth rate. During the coagulation period, the fractal dimension of floc aggregates increased in the first minutes and then decreased and larger flocs generally had smaller fractal dimensions. The floc strength, which was assessed by the relationship of floc diameter and velocity gradient, decreased with the increase of coagulation pH. Flocs formed at pH 4.00 had better recovery capability when exposed to lower shear forces, while flocs formed at neutral and alkaline conditions had better performance under higher shear forces.

Effect of OH-/Al3+ and Si/Al molar ratios on the coagulation performance and residual Al speciation during surface water treatment with poly-aluminum-silicate-chloride (PASiC).

Coagulation performance, mechanism of poly-aluminum-silicate-chloride (PASiC) and residual Al speciation in the effluent with respect to a specific surface water treatment in China were comprehensively investigated in this study. The impact of OH(-)/Al(3+) and Si/Al molar ratios on the coagulation performance, mechanism and residual Al speciation of PASiC in surface water treatment was discussed as a function of coagulant dosage. It was intended to provide an insight into the relationship between coagulation performance and residual Al. Experimental results revealed that when OH(-)/Al(3+) molar ratio = 2.00 and Si/Al molar ratio = 0.0500 in PASiC coagulant, PASiC exhibited beneficial coagulation property and relatively lower content of residual Al. Surface bridging and entrapment was more effective compared with charge neutralization during the specific surface water treatment. The majority of residual Al in the effluent existed in the form of insoluble suspended or particulate Al. Dissolved organically bound Al was almost the major speciation in dissolved Al and dissolved inorganically bound monomeric Al was the only component in dissolved monomeric Al. Al in PASiC remained abundant at lower dosages and residual Al concentration could be effectively reduced at the dosages of 12.0-15.0mg/L as Al.

[Treatment of simulated produced wastewater from polymer flooding in oil production using dithiocarbamate-type flocculant].

A dithiocarbamate flocculant, DTC (T403), was prepared by the reaction of amine-terminated polyoxypropane-ether compound known as Jeffamine-T403 and carbon disulfide in alkaline solution. The oil removal efficiency of DTC (T403) for simulated produced wastewater from polymer flooding in oil production was studied by Jar-test. The effect of the dosage of DTC (T403), hydrolyzed polyacrylamide (HPAM), Fe2+ and Fe3+ ions, and pH on the oil removal efficiency of DTC (T403) was investigated. The results showed that the chelate polymer formed by DTC (T403) and Fe2+ ion has good oil removal performance by net capturing mechanism. HPAM had a negative effect on oil removal efficiency of DTC (T403). For the treatment of the simulated wastewater containing 0-900 mg/L of HPAM and 300 mg/L of oil, the residual oil concentrations in water samples decreased below 10 mg/L when the dosage of Fe2+ and DTC (T403) was 10 mg/L and 25 mg/L, respectively. The oil removal efficiency of DTC (T403) was affected by pH and good oil removal efficiency was obtained when the pH was below 7.5. DTC (T403) is appropriate for the treatment of oily wastewater containing Fe2+ ion.

[Municipal wastewater treatment using a composite flocculant made of polyaluminum chloride and polydimethyldiallyammonium chloride].

A composite flocculant (denoted JYF-1), made of polyaluminum chloride (PAC) and polydimethyldiallyammonium chloride (PDMDAAC), was used in jar-tests to simulate the chemically enhanced primary treatment (CEPT) for municipal wastewater. Removal of particles, organic compounds, nitrogen and phosphorus in wastewater was investigated, and the effects of pH and surface overflow rate (SOR) on flocculation were also examined. Electrical charge and distribution of particles in wastewater were analyzed before and after flocculation. Furthermore, the flocculation mechanism and application of JYF-1 in CEPT were discussed. The results demonstrate that JYF-1 performs better than PAC under a wide pH and SOR range. When 8 mg x L(-1) JYF-1 is added, 76.72% COD and 64.31% soluble COD (SCOD) can be removed. About 90% soluble TP (STP), 80% TP and 20% TN can be removed by addition of 12 mg x L(-1) JYF-1. After flocculation, the BOD/COD ratio increases from 0.23 to 0.53, which indicates the biodegradation ability of wastewater is improved. It can be concluded that JYF-1 is a high-efficiency low-cost flocculant, which can improve outlet water quality and produce less sludge without changing the existing equipments and treating process in sewage plants.

[Flocculation of wastewater produced in polymer flooding].

The particle size distribution, flocculation index, Zeta potential and oil content in water were tested during flocculation of wastewater produced in polymer flooding of oil field. CHP-03, a mixture of a modified pluronic demulsifer and a weak-cationic flocculant, was used to improve the efficiency of oil droplet collision. The effects of polyacrylamide, temperature, and mixing speed on flocculation dynamics are discussed. Polyacrylamide affects the oil-water separation by obstructing the aggregation of small oil droplets, but electrostatic repulsion is not a major factor in preventing flocculation. The local temperature has a limited effect on flocculation. Fast mixing favors the growth of flocs, while slow mixing is suitable for the coalescence of oil droplets. Addition of 250 mg/L CHP-03 can enhance the oil-water separation and remove 92% of the oil content.