The comprehensive effects of aluminum oxide nanoparticles on the physiology of freshwater microalga Scenedesmus obliquus and it's phycoremediation performance for the removal of sulfacetamide.

Nanoparticles are inevitable byproducts of modern industry. However, the environmental impacts arising from industrial applications of nanoparticles are largely under-reported. This study evaluated the ecotoxicological effects of aluminum oxide nanoparticles (Al2O3NP) and its influence on sulfacetamide (SA) biodegradation by a freshwater microalga, Scenedesmus obliquus. Although Al2O3NP showed limited toxicity effect on S. obliquus, we observed the toxicity attenuation aspect of Al2O3NP in a mixture of sulfacetamide on microalgae. The addition of 100 mg L-1 of Al2O3NP and 1 mg L-1 of SA reduced total chlorophyll by 23.3% and carotenoids by 21.6% in microalgal compared to control. The gene expression study demonstrated that ATPF0C, Lhcb1, HydA, and psbA genes responsible for ATP synthesis and the photosynthetic system were significantly downregulated, while the Tas gene, which plays a major role in biodegradation of organic xenobiotic chemicals, was significantly upregulated at 1 and 100 mg L-1 of Al2O3NP. The S. obliquus removed 16.8% of SA at 15 mg L-1 in 14 days. However, the removal was slightly enhanced (18.8%) at same concentration of SA in the presence of 50 mg L-1 Al2O3NP. This result proves the stability of sulfacetamide biodegradation capacity of S. obliquus in the presence of Al2O3NP co-contamination. The metabolic analysis showed that SA was degraded into simpler byproducts such as sulfacarbamide, sulfaguanidine, sulfanilamide, 4-(methyl sulfonyl)aniline, and N-hydroxy-benzenamine which have lower ecotoxicity than SA, demonstrating that the ecotoxicity of sulfacetamide has significantly decreased after the microalgal degradation, suggesting the environmental feasibility of microalgae-mediated wastewater technology. This study provides a deeper understanding of the impact of nanoparticles such as Al2O3NP on aquatic ecosystems.

Marine Alga Ulva fasciata-Derived Molecules for the Potential Treatment of SARS-CoV-2: An In Silico Approach.

SARS-CoV-2 is the causative agent of the COVID-19 pandemic. This in silico study aimed to elucidate therapeutic efficacies against SARS-CoV-2 of phyco-compounds from the seaweed, Ulva fasciata. Twelve phyco-compounds were isolated and toxicity was analyzed by VEGA QSAR. Five compounds were found to be nonmutagenic, noncarcinogenic and nontoxic. Moreover, antiviral activity was evaluated by PASS. Binding affinities of five of these therapeutic compounds were predicted to possess probable biological activity. Fifteen SARS-CoV-2 target proteins were analyzed by the AutoDock Vina program for molecular docking binding energy analysis and the 6Y84 protein was determined to possess optimal binding affinities. The Desmond program from Schrödinger's suite was used to study high performance molecular dynamic simulation properties for 3,7,11,15-Tetramethyl-2-hexadecen-1-ol-6Y84 for better drug evaluation. The ligand with 6Y84 had stronger binding affinities (-5.9 kcal/mol) over two standard drugs, Chloroquine (-5.6 kcal/mol) and Interferon α-2b (-3.8 kcal/mol). Swiss ADME calculated physicochemical/lipophilicity/water solubility/pharmacokinetic properties for 3,7,11,15-Tetramethyl-2-hexadecen-1-ol, showing that this therapeutic agent may be effective against SARS-CoV-2.

Biodiesel production potential of microalgae, cultivated in acid mine drainage and livestock wastewater.

The adaptability and biofuel production potential of two strains of microalgae isolated and cultivated in livestock wastewater effluent (LWE) with acid mine drainage (AMD) were investigated. The isolated strains of microalgae from samples obtained from LWE and AMD, two microalgal strains (Nephroselmis sp. KGE2 and Autodesmus obliquus KGE17) were selected based on their growth rate and lipid productivity. The dry cell weight of Nephroselmis sp. KGE2 and Autodesmus obliquus KGE17 after 20 days of cultivation in AMD increased from 0.05 to 0.59 g/L and from 0.05 to 0.55 g/L, respectively. These findings revealed a significant accumulation of fatty acids with increasing AMD content. Nephroselmis sp. KGE2 in LWE with 5% AMD demonstrated a higher growth rate (0.59 ± 0.03 g/L) and fatty acid production (401.5 ± 47.3 mg/L) than Autodesmus obliquus KGE17 with 5% AMD. Additionally, Nephroselmis sp. KGE2 had C16-C18 fatty acid content (92.4%) in LWE with AMD. Biodiesel produced from Nephroselmis sp. KGE2 had a higher cetane number (52.31) and iodine value (88.26 g I2/100 g oil). Consequently, Nephroselmis sp. KGE2 can be considered a potential candidate for biodiesel production using AMD as an iron source.

Efficient removal of formaldehyde using metal-biochar derived from acid mine drainage sludge and spent coffee waste.

A novel metal-biochar (Biochar/AMDS) composite were fabricated by co-pyrolysis of spent coffee waste (SCW)/acid mine drainage sludge (AMDS), and their effective application in adsorptive removal of air pollutants such as formaldehyde in indoor environments was evaluated. The physicochemical characteristics of Biochar/AMDS were analyzed using SEM/EDS, XRF, XRD, BET, and FTIR. The characterization results illustrated that Biochar/AMDS had the highly porous structure, carbonaceous layers, and heterogeneous Fe phases (hematite, metallic Fe, and magnetite). The fixed-bed column test showed that the removal of formaldehyde by Biochar/AMDS was 18.4-fold higher than that by metal-free biochar (i.e., SCW-derived biochar). Changing the ratio of AMDS from 1:6 to 1:1 significantly increased the adsorption capacity for formaldehyde from 1008 to 1811 mg/g. In addition, thermal treatment of used adsorbent at 100 °C effectively restored the adsorptive function exhausted during the column test. These results provide new insights into the fabrication of practical, low-cost and ecofriendly sorbent for formaldehyde.

Efficient removal of arsenic by strategically designed and layer-by-layer assembled PS@+rGO@GO@Fe3O4 composites.

The PS@+rGO@GO@Fe3O4 (PG-Fe3O4) hybrid composites for Arsenic removal were successfully fabricated and well dispersed using layer-by-layer assembly and a hydrothermal method. The PG-Fe3O4 hybrid composites were composed of uniformly coated Fe3O4 nanoparticles on graphene oxide layers with water flow space between 3D structures providing many contact area and adsorption sites for Arsenic adsorption. The PG-Fe3O4 hybrid composite has large surface adsorption sites and exhibits high adsorption capacities of 104 mg/g for As (III) and 68 mg/g for As (V) at 25 °C and pH 7 comparison with pure Fe3O4 and P-Fe3O4 samples.

7-Ketocholesterol induces the reduction of KCNMB1 in atherosclerotic blood vessels.

Hypertension is a high-risk symptom in atherosclerotic patients, and vascular rigidity is one of the main factors leading to hypertension. ß1-Subunit of BKCa channel (KCNMB1; MaxiKß1) has been reported as a modulator of vascular flexibility. To determine the relationship between atherosclerosis and KCNMB1, we studied some atherogenic factors affecting vascular tone. Blood of atherosclerotic patients shows increased concentration of 7-ketocholesterol (7K), which has been studied as a harmful lipid to blood vessels. Our data showed that KCNMB1 was significantly down-regulated in the presence of 7K, in a dose-/time-dependent manner in vascular smooth muscle cells (VSMCs). And, the reduction of KCNMB1 was confirmed in cell images of 7K-stimulated VSMCs and in vessel tissue images of ApoE knock-out mice. To determine whether aryl hydrocarbon receptor (AhR) was involved in the reduction of KCNMB1 by 7K-stimulation, protein level of AhR was analyzed by Western blot. Our data showed that the reduction of KCNMB1 was modulated through the AhR pathway. In conclusion, results of our study suggest that 7K induces the reduction of KCNMB1 through the AhR pathway.

Reference and counter electrode positions affect electrochemical characterization of bioanodes in different bioelectrochemical systems.

The placement of the reference electrode (RE) in various bioelectrochemical systems is often varied to accommodate different reactor configurations. While the effect of the RE placement is well understood from a strictly electrochemistry perspective, there are impacts on exoelectrogenic biofilms in engineered systems that have not been adequately addressed. Varying distances between the working electrode (WE) and the RE, or the RE and the counter electrode (CE) in microbial fuel cells (MFCs) can alter bioanode characteristics. With well-spaced anode and cathode distances in an MFC, increasing the distance between the RE and anode (WE) altered bioanode cyclic voltammograms (CVs) due to the uncompensated ohmic drop. Electrochemical impedance spectra (EIS) also changed with RE distances, resulting in a calculated increase in anode resistance that varied between 17 and 31 Ω (-0.2 V). While WE potentials could be corrected with ohmic drop compensation during the CV tests, they could not be automatically corrected by the potentiostat in the EIS tests. The electrochemical characteristics of bioanodes were altered by their acclimation to different anode potentials that resulted from varying the distance between the RE and the CE (cathode). These differences were true changes in biofilm characteristics because the CVs were electrochemically independent of conditions resulting from changing CE to RE distances. Placing the RE outside of the current path enabled accurate bioanode characterization using CVs and EIS due to negligible ohmic resistances (0.4 Ω). It is therefore concluded for bioelectrochemical systems that when possible, the RE should be placed outside the current path and near the WE, as this will result in more accurate representation of bioanode characteristics.

A two-stage microbial fuel cell and anaerobic fluidized bed membrane bioreactor (MFC-AFMBR) system for effective domestic wastewater treatment.

Microbial fuel cells (MFCs) are a promising technology for energy-efficient domestic wastewater treatment, but the effluent quality has typically not been sufficient for discharge without further treatment. A two-stage laboratory-scale combined treatment process, consisting of microbial fuel cells and an anaerobic fluidized bed membrane bioreactor (MFC-AFMBR), was examined here to produce high quality effluent with minimal energy demands. The combined system was operated continuously for 50 days at room temperature (∼25 °C) with domestic wastewater having a total chemical oxygen demand (tCOD) of 210 ± 11 mg/L. At a combined hydraulic retention time (HRT) for both processes of 9 h, the effluent tCOD was reduced to 16 ± 3 mg/L (92.5% removal), and there was nearly complete removal of total suspended solids (TSS; from 45 ± 10 mg/L to <1 mg/L). The AFMBR was operated at a constant high permeate flux of 16 L/m(2)/h over 50 days, without the need or use of any membrane cleaning or backwashing. Total electrical energy required for the operation of the MFC-AFMBR system was 0.0186 kWh/m(3), which was slightly less than the electrical energy produced by the MFCs (0.0197 kWh/m(3)). The energy in the methane produced in the AFMBR was comparatively negligible (0.005 kWh/m(3)). These results show that a combined MFC-AFMBR system could be used to effectively treat domestic primary effluent at ambient temperatures, producing high effluent quality with low energy requirements.

Continuous augmentation of anaerobic digestion with electroactive microorganisms: Performance and stability.

The performance and stability of a bioelectrochemical anaerobic digester (BeAD), continuously augmented with electroactive microorganisms (EAMs), were investigated. The BeAD showcased superior performance, sustaining the high COD removal efficiency and methane production rate of 76.5 % and 0.67 L/(L.d), respectively, in a stable state. Prominently, it exhibited remarkable resilience under hydraulic and organic shock loads, adeptly recuperating from disturbances up to 1000 % of its stable condition. This resilience of up to 300 % shock load was driven by increased levels of electron transport components such as quinones and riboflavins, which act as electron shuttles. However, after extreme shock exposures from 500 % to 1000 %, despite the spike in inhibitory by-products such as humic acids and ammonia, the upregulation of the mtr complex was pivotal in recovering and sustaining methane production. These insights emphasize the BeAD's capability to bolster both performance and stability, thereby providing a potent strategy for practical application of bioelectrochemical systems.

Artificial neural network modeling for the oxidation kinetics of divalent manganese ions during chlorination and the role of arsenite ions in the binary/ternary systems.

This study investigated the coexistence and contamination of manganese (Mn(II)) and arsenite (As(III)) in groundwater and examined their oxidation behavior under different equilibrating parameters, including varying pH, bicarbonate (HCO3-) concentrations, and sodium hypochlorite (NaClO) oxidant concentrations. Results showed that if the molar ratio of NaClO: As(III) was >1, the oxidation of As(III) could be achieved within a minute with an extremely high oxidation rate of 99.7 %. In the binary system, the removal of As(III) prevailed over Mn(II). The As(III) oxidation efficiency increased from 59.8 ± 0.6 % to 70.8 ± 1.9 % when pH rose from 5.7 to 8.0. The oxidation reaction between As(III) and NaClO releases H+ ions, decreasing the pH from 6.77 to 6.19 and reducing the removal efficiency of Mn(II). The presence of HCO3- reduced the oxidation rate of Mn(II) from 63.2 % to 13.9 % within four hours. Instead, the final oxidation rate of Mn(II) increased from 68.1 % to 87.7 %. This increase can be attributed to HCO3- ions competing with the free Mn(II) for the adsorption sites on the sediments, inhibiting the formation of H+. Moreover, kinetic studies revealed that the oxidation reaction between Mn(II) and NaClO followed first-order kinetics based on their R2 values. The significant factors affecting the Mn(II) oxidation efficiency were the initial concentration of NaClO and pH. Applying an artificial neural network (ANN) model for data analysis proved to be an effective tool for predicting Mn(II) oxidation rates under different experimental conditions. The actual Mn(II) oxidation data and the predicted values obtained from the ANN model showed significant consistency. The training and validation data sets yielded R2 values of 0.995 and 0.992, respectively. Moreover, the ANN model highlights the importance of pH and NaClO concentrations in influencing the oxidation rate of Mn(II).

Domestic wastewater treatment using multi-electrode continuous flow MFCs with a separator electrode assembly design.

Treatment of domestic wastewater using microbial fuel cells (MFCs) will require reactors with multiple electrodes, but this presents unique challenges under continuous flow conditions due to large changes in the chemical oxygen demand (COD) concentration within the reactor. Domestic wastewater treatment was examined using a single-chamber MFC (130 mL) with multiple graphite fiber brush anodes wired together and a single air cathode (cathode specific area of 27 m(2)/m(3)). In fed-batch operation, where the COD concentration was spatially uniform in the reactor but changed over time, the maximum current density was 148 ± 8 mA/m(2) (1,000 Ω), the maximum power density was 120 mW/m(2), and the overall COD removal was >90 %. However, in continuous flow operation (8 h hydraulic retention time, HRT), there was a 57 % change in the COD concentration across the reactor (influent versus effluent) and the current density was only 20 ± 13 mA/m(2). Two approaches were used to increase performance under continuous flow conditions. First, the anodes were separately wired to the cathode, which increased the current density to 55 ± 15 mA/m(2). Second, two MFCs were hydraulically connected in series (each with half the original HRT) to avoid large changes in COD among the anodes in the same reactor. The second approach improved current density to 73 ± 13 mA/m(2). These results show that current generation from wastewaters in MFCs with multiple anodes, under continuous flow conditions, can be improved using multiple reactors in series, as this minimizes changes in COD in each reactor.

Assessment of effectiveness in stabilization/solidification of arsenic-contaminated soil: long-term leaching test and geophysical measurement.

This study focused on evaluating the effectiveness of stabilizer/binding agents in immobilizing arsenic (As) in contaminated soil using both geochemical and geophysical monitoring methods. The effluent from the stabilizer/binding agent's application and control columns was analyzed, and the status of the columns was monitored using electrical resistivity (ER) and induced polarization (IP) methods. As stabilizers/binder, acid mine drainage sludge (AMDS) and steel slag (SS) were used, which delayed As and Ca leaching time and significantly reduced As leaching amount. Determination coefficients for As and Fe leaching exhibited elevated values (control column, R2 = 0.955; AMDS column, R2 = 0.908; and SS column, R2 = 0.833). A discernible decline in the concentration of leached Fe was accompanied by a corresponding reduction in IP. The determination coefficients correlating IP and Fe leaching remained substantial (control column, R2 = 0.768; AMDS column, R2 = 0.807; and SS column, R2 = 0.818). Such IP measurements manifest as instrumental tools in monitoring and assessing the retention capacity of applied stabilizer/binding agents in As-affected soils, thereby furnishing crucial data for the enduring surveillance of stabilization/solidification locales. This research posits a swift and continuous monitoring method for solidification/stabilization locales in situ.

Evaluation of pyrite/sodium hypochlorite for activating purification of arsenic from fractured-bedrock groundwater.

In this work, the effectiveness of pyrite/sodium hypochlorite (FeS2/NaClO) treatment to eliminate arsenic (As) from fractured-bedrock groundwater via oxidative adsorption was evaluated. The As concentration in the tested reactors decreased sharply during the initial 5 min, as the addition of NaClO effectively increased the As removal efficiency, attaining 98.6% removal within 60 min in the presence of 0.05 M NaClO. There was no coexisting anion effect (Cl-, CO3-, HCO3-, NO3-, and F-) on the As removal capacity of FeS2/NaClO, except for the PO43- which resulted in less removal of As. X-ray spectroscopy analysis of As(III)-sorbed FeS2 surfaces revealed that a portion of As(III) was oxidized into As(V) during the adsorption process. Scanning electron microscopy-energy-dispersive spectrometer results of FeS2 exhibited the distribution of adsorbed As on the newly formed iron (oxy) hydroxide surfaces, with an As element ratio of 1.27%. A continuous flow-bed column study further demonstrated the efficiency of FeS2/NaClO treatment to lower the contamination level of As at the removal rates of 0.66-3.02 mg/L·day for 160 h. These results suggest that FeS2/NaClO treatment can be considered an effective strategy for removing As in groundwater of bedrock aquifers.

Morphology and stability of mineralized carbon influenced by magnesium ions.

Ex situ mineralization of CO2 is a promising technology that employs Ca- and Mg-rich industrial wastes but it simultaneously produces end products. Although Mg is a major mineralization source, it can adversely impact carbonate precipitation and crystal stability during co-precipitation in combination with Ca2+. In this study, the effects of Mg2+ ions on the mineralization process and its products were investigated using precipitates formed at different aqueous concentrations of Mg2+. The final phases of the precipitates were quantitatively evaluated at the end of each process. The alterations undergone by the calcite crystals, which constituted the dominant carbonate phase in each experiment, were analyzed using a sophisticated crystallographic approach. Aragonite was detected at high Mg2+ concentrations (Mg2+/Ca2+ ratio of 2.00), although brucite was the sole phase of the Mg crystal. The increase in Mg2+ ion concentration induced the formation of an amorphous solid. The results revealed that a drastic transformation of the calcite lattice occurred when the ratio of Mg2+/Ca2+ exceeded 1.00, agreeing with the shifts observed in the calcite structure upon comparing the precipitates formed at the Mg2+/Ca2+ ratios of 1.00 and 2.00, wherein microstrain and crystallite sizes changed from 0.040 and 55.33 nm to 0.1533 and 12.35 nm, respectively. At a Mg2+/Ca2+ ratio of 2.00, 6.51% of the Ca2+ ions in the calcite structure were substituted by Mg2+, increasing the surface energy of the crystal and the solubility of the carbonate. Therefore, Mg2+ is a potential hindrance that can impede the precipitation of carbonates and increase instability at certain concentrations.

Fluoride occurrence in environment, regulations, and remediation methods for soil: A comprehensive review.

Fluoride, a naturally occurring chemical element, is largely insoluble in soils. More than 90% of the fluoride in soil is bound to soil particles and is unable to be dissolved. As part of the soil, fluoride is predominantly located in the colloid or clay fraction of the soil, and the movement of fluoride is strongly affected by the sorption capacity of the soil, which is affected by pH, the type of soil sorbent present, and the salinity. The Canadian Council of Ministers of the Environment soil quality guideline for fluoride in soils under a residential/parkland land use scenario is 400 mg/kg. In this review, we focus on fluoride contamination in soil and subsurface environments, and the various sources of fluorides are discussed in detail. The average fluoride concentration in soil in different countries and their regulations for soil and water are comprehensively reviewed. In this article, the latest advances in defluoridation methods are highlighted and the importance of further research addressing efficient and cost-effective methods to remediate fluoride contamination in soil is critically discussed. Methods used to mitigate fluoride risks by removing fluoride from the soil are presented. We strongly recommend that regulators and soil chemists in all countries explore opportunities to improve defluoridation methods and consider adopting more stringent regulations for fluoride in soil depending on geologic conditions.

Fate of five bisphenol derivatives in Chlamydomonas mexicana: Toxicity, removal, biotransformation and microalgal metabolism.

Bisphenols (BPs) are recognized as emerging contaminants because of their estrogenic properties and frequent occurrence in environmental matrices. Here, we evaluated the toxic effects of five common BPs on freshwater microalga Chlamydomonas mexicana and removal of the BPs by the alga. Bisphenols -AF (BPAF), -B (BPB), and -Z (BPZ) (96 h, EC50 1.78-12.09 mg·L-1) exhibited higher toxicity to C. mexicana compared to bisphenol -S (BPS) and -F (BPF) (96 h, EC50 30.53-85.48 mg·L-1). In contrast, the mixture of BPs exhibited acute toxicity (96 h, EC50 8.07 mg·L-1). After 14 days, C. mexicana had effectively removed 61%, 99%, 55%, 87%, and 89% of BPS, BPF, BPAF, BPB, and BPZ, respectively, at 1 mg L-1. The biotransformed products of all five BPs were analyzed using UHPLC QTOF, and their toxicity was predicted. All biotransformed products were observed to be less toxic than the parent compounds. The fatty acid composition of C. mexicana after exposure to the BP mixture was predominantly palmitic acid (34.14%), followed by oleic acid (18.9%), and γ-linolenic acid (10.79%). The results provide crucial information on the ecotoxicity of these five BPs and their removal by C. mexicana; the resulting biomass is a potential feedstock for producing biodiesel.

Molecular characterization of azoreductase and its potential for the decolorization of Remazol Red R and Acid Blue 29.

Azoreductase is a reductive enzyme that efficiently biotransformed textile azo dyes. This study demonstrated the heterologous overexpression of the azoreductase gene in Escherichia coli for the effective degradation of Remazol Red-R and Acid-Blue 29 dyes. The AzK gene of Klebsiella pneumoniae encoding a ≈22 kDa azoreductase enzyme was cloned into the pET21+C expression vector. The inoculum size of 1.5%, IPTG concentration of 0.5 mM, and incubation time of 6 h were optimized by response surface methodology a statistical tool. The crude extract showed 76% and 74%, while the purified enzyme achieved 94% and 93% decolorization of RRR and AB-29, respectively in 0.3 h. The reaction kinetics showed that RRR had a Km and Vmax value of 0.058 mM and 1416 U mg-1, respectively at an NADH concentration of 10 mM. HPLC and GC-MS analyses showed that RRR was effectively bio-transformed by azoreductase to 2-[3-(hydroxy-amino) benzene-1-sulfonyl and AB-29 to aniline and 3-nitrosoaniline. This study explored the potential of recombinant azoreductase isolated from K. pneumoniae in the degradation of toxic textile azo dyes into less toxic metabolites.

Polyol-mediated zinc oxide nanoparticles using the refluxing method as an efficient photocatalytic and antimicrobial agent.

Nanomaterials have attracted more curiosity recently because of their wide-ranging application in environmental remediation and electronic devices. The current study focuses on zinc oxide nanoparticles' (ZnO NPs) simple production, characterization, and applications in several fields, including medicinal and photocatalytic degradation of dyes. The non-aqueous-based reflux method is helpful for ZnO NP synthesis; the procedure involves refluxing zinc acetate dihydrate precursor in ethylene glycol for 3 hours in the absence of sodium acetate, in which the refluxing rate and the cooling rate are optimized to get the desired phase, and the unique morphology of polyol-mediated ZnO NPs; it has been achieved using the capping agent TBAB (tetra-butyl ammonium bromide) and precursor zinc acetate dihydrate. UV-Vis, FTIR, XRD, and FESEM structurally characterized polyol-mediated ZnO-NPs. The results show that the material is pure and broadly aggregated into spherical nanoparticles with an average particle size of 18.09 nm. According to XRD analysis, heat annealing made the crystallites more prominent and favored a monocrystalline state. These results and the low cost of making polyol-mediated ZnO NPs demonstrate photocatalytic and antimicrobial properties.

Hypoxia-inducible factor-1α regulates KCNMB1 expression in human pulmonary artery smooth muscle cells.

Previously, we observed that hypoxia increases the expression of the ß1-subunit (KCNMB1) of the calcium-sensitive potassium channel (BK(Ca)). Herein, we elucidate the mechanism whereby hypoxia increases KCNMB1 expression in human pulmonary artery smooth muscle cells (hPASMC). In response to hypoxia, the expression of both the transcription factor hypoxia-inducible factor 1-α (HIF-1α) and KCNMB1 are increased. Knockdown of HIF-1α using a shRNA plasmid blocked the hypoxic induction of KCNMB1 expression. Chromatin immunoprecipitation (ChIP) demonstrated HIF-1α binding to three discrete regions of the human KCNMB1 promoter known to contain hypoxia response elements (HREs). A KCNMB1 promoter reporter assay combined with site-directed mutagenesis identified two adjacent HREs located between -3,540 bp and -3,311 bp that are essential for the hypoxic induction of KCNMB1 promoter activity. Furthermore, additional ChIP assays demonstrated recruitment of the HIF-1α transcriptional coactivator, p300, to this same promoter region. Treatment of hPASMC with the histone deacetylase inhibitor, trichostatin, prolonged the increase in KCNMB1 observed with hypoxia, suggesting that alterations in chromatin remodeling function to limit the hypoxic induction of KCNMB1. Finally, KCNMB1 knockdown potentiated the hypoxia-induced increase in cytosolic calcium in hPASMC, highlighting the contribution of the ß1-subunit in modulating vascular SMC tone in response to acute hypoxia. In conclusion, HIF-1α increases KCNMB1 expression in response to hypoxia in hPASMC by binding to two HREs located at -3,540 to -3,311 of the KCNMB1 promoter. We speculate that selective modulation of KCNMB1 expression may serve as a novel therapeutic approach to address diseases characterized by an increase in vascular tone.

A multi-electrode continuous flow microbial fuel cell with separator electrode assembly design.

Scaling up microbial fuel cells (MFCs) requires the development of compact reactors with multiple electrodes. A scalable single chamber MFC (130 mL), with multiple graphite fiber brush anodes and a single air-cathode cathode chamber (27 m2/m3), was designed with a separator electrode assembly (SEA) to minimize electrode spacing. The maximum voltage produced in fed-batch operation was 0.65 V (1,000 Ω) with a textile separator, compared to only 0.18 V with a glass fiber separator due to short-circuiting by anode bristles through this separator with the cathode. The maximum power density was 975 mW/m2, with an overall chemical oxygen demand (COD) removal of >90% and a maximum coulombic efficiency (CE) of 53% (50 Ω resistor). When the reactor was switched to continuous flow operation at a hydraulic retention time (HRT) of 8 h, the cell voltage was 0.21 ± 0.04 V, with a very high CE = 85%. Voltage was reduced to 0.13 ± 0.03 V at a longer HRT = 16 h due to a lower average COD concentration, and the CE (80%) decreased slightly with increased oxygen intrusion into the reactor per amount of COD removed. Total internal resistance was 33 Ω, with a solution resistance of 2 Ω. These results show that the SEA type MFC can produce stable power and a high CE, making it useful for future continuous flow treatment using actual wastewaters.