Synergistic material-microbe interface toward deeper anaerobic defluorination.

Per- and polyfluoroalkyl substances (PFAS), particularly the perfluorinated ones, are recalcitrant to biodegradation. By integrating an enrichment culture of reductive defluorination with biocompatible electrodes for the electrochemical process, a deeper defluorination of a C6-perfluorinated unsaturated PFAS was achieved compared to the biological or electrochemical system alone. Two synergies in the bioelectrochemical system were identified: i) The in-series microbial-electrochemical defluorination and ii) the electrochemically enabled microbial defluorination of intermediates. These synergies at the material-microbe interfaces surpassed the limitation of microbial defluorination and further turned the biotransformation end products into less fluorinated products, which could be less toxic and more biodegradable in the environment. This material-microbe hybrid system brings opportunities in the bioremediation of PFAS driven by renewable electricity and warrants future research on mechanistic understanding of defluorinating and electroactive microorganisms at the material-microbe interface for system optimizations.

Electron bifurcation and fluoride efflux systems implicated in defluorination of perfluorinated unsaturated carboxylic acids by Acetobacterium spp.

Enzymatic cleavage of C─F bonds in per- and polyfluoroalkyl substances (PFAS) is largely unknown but avidly sought to promote systems biology for PFAS bioremediation. Here, we report the reductive defluorination of α, ß-unsaturated per- and polyfluorocarboxylic acids by Acetobacterium spp. The microbial defluorination products were structurally confirmed and showed regiospecificity and stereospecificity, consistent with their formation by enzymatic reactions. A comparison of defluorination activities among several Acetobacterium species indicated that a functional fluoride exporter was required for the detoxification of the released fluoride. Results from both in vivo inhibition tests and in silico enzyme modeling suggested the involvement of enzymes of the flavin-based electron-bifurcating caffeate reduction pathway [caffeoyl-CoA reductase (CarABCDE)] in the reductive defluorination. This is a report on specific microorganisms carrying out enzymatic reductive defluorination of PFAS, which could be linked to electron-bifurcating reductases that are environmentally widespread.

Aerobic Biotransformation and Defluorination of Fluoroalkylether Substances (ether PFAS): Substrate Specificity, Pathways, and Applications.

Fluoroalkylether substances (ether PFAS) constitute a large group of emerging PFAS with uncertain environmental fate. Among them, GenX is the well-known alternative to perfluorooctanoic acid and one of the six proposed PFAS to be regulated by the U.S. Environmental Protection Agency. This study investigated the structure-biodegradability relationship for 12 different ether PFAS with a carboxylic acid headgroup in activated sludge communities. Only polyfluorinated ethers with at least one -CH2- moiety adjacent to or a C=C bond in the proximity of the ether bond underwent active biotransformation via oxidative and hydrolytic O-dealkylation. The bioreactions at ether bonds led to the formation of unstable fluoroalcohol intermediates subject to spontaneous defluorination. We further demonstrated that this aerobic biotransformation/defluorination could complement the advanced reduction process in a treatment train system to achieve more cost-effective treatment for GenX and other recalcitrant perfluorinated ether PFAS. These findings provide essential insights into the environmental fate of ether PFAS, the design of biodegradable alternative PFAS, and the development of cost-effective ether PFAS treatment strategies.

Substantial defluorination of polychlorofluorocarboxylic acids triggered by anaerobic microbial hydrolytic dechlorination.

Chlorinated polyfluorocarboxylic acids (Cl-PFCAs) derived from the widely used chlorotrifluoroethylene (CTFE) polymers and oligomers may enter and influence the aquatic environment. Here, we report significant defluorination of Cl-PFCAs by an anaerobic microbial community via novel pathways triggered by anaerobic microbial dechlorination. Cl-PFCAs first underwent microbial reductive, hydrolytic, and eliminative dechlorination, and it was the hydrolytic dechlorination that led to significant spontaneous defluorination. Hydrolytic dechlorination was favored with increased Cl-substitutions. An isolated, highly enriched anaerobic defluorinating culture was dominated by two genomes closest to Desulfovibrio aminophilus and Sporomusa sphaeroides, both of which exhibited active defluorination of CTFE tetramer acid. It implies the critical role played by anaerobic non-respiratory hydrolytic dechlorination in the fate of chlorinated polyfluoro-chemicals in natural and engineered water environments. The greatly enhanced biodegradability by Cl-substitutions also sheds light on the design of cost-effective treatment biotechnologies, as well as alternative PFAS that are readily biodegradable and less toxic.

Electron-bifurcation and fluoride efflux systems in Acetobacterium spp. drive defluorination of perfluorinated unsaturated carboxylic acids.

Enzymatic cleavage of C-F bonds in per- and polyfluoroalkyl substances (PFAS) is largely unknown but avidly sought to promote systems biology for PFAS bioremediation. Here, we report the reductive defluorination of α, ß-unsaturated per- and polyfluorocarboxylic acids by Acetobacterium spp. Two critical molecular features in Acetobacterium species enabling reductive defluorination are (i) a functional fluoride efflux transporter (CrcB) and (ii) an electron-bifurcating caffeate reduction pathway (CarABCDE). The fluoride transporter was required for detoxification of released fluoride. Car enzymes were implicated in defluorination by the following evidence: (i) only Acetobacterium spp. with car genes catalyzed defluorination; (ii) caffeate and PFAS competed in vivo ; (iii) models from the X-ray structure of the electron-bifurcating reductase (CarC) positioned the PFAS substrate optimally for reductive defluorination; (iv) products identified by 19 F-NMR and high-resolution mass spectrometry were consistent with the model. Defluorination biomarkers identified here were found in wastewater treatment plant metagenomes on six continents.

Performance enhancement, membrane fouling mitigation and eco-friendly strategy by electric field coupled membrane bioreactor for treating mariculture wastewater.

An eco-friendly strategy for mariculture wastewater treatment using an electric field attached membrane bioreactor (E-MBR) was evaluated and compared with a conventional membrane bioreactor (C-MBR). The removal efficiencies of total nitrogen (TN) and chemical oxygen demand (COD) increased significantly and the membrane fouling rate reduced by 44.8% in the E-MBR. The underlying mechanisms included the enriched nitrifiers and denitrifiers, the enhanced salinity-resistance, the increased activities and upregulated genes of key enzymes involved in nitrification and denitrification for improving the performance of mariculture wastewater treatment, and the enriched extracellular polymeric substance (EPS)-degrading genera, the downregulated EPS biosynthesis genes, the repressed biofilm-forming bacteria, the enhanced zeta potential absolute value and the generated H2O2 for membrane fouling mitigation by electrical stimulation. Compared with the C-MBR, the energy consumption, carbon emissions, and nitrogen footprint were reduced. These findings provide novel insights into mariculture wastewater treatment using an applied electric field.

Microbial Defluorination of Unsaturated Per- and Polyfluorinated Carboxylic Acids under Anaerobic and Aerobic Conditions: A Structure Specificity Study.

The recently discovered microbial reductive defluorination of two C6 branched and unsaturated fluorinated carboxylic acids (FCAs) provided valuable insights into the environmental fate of per- and polyfluoroalkyl substances (PFASs) and potential bioremediation strategies. However, a systematic investigation is needed to further demonstrate the role of C═C double bonds in the biodegradability of unsaturated PFASs. Here, we examined the structure-biodegradability relationships of 13 FCAs, including nine commercially available unsaturated FCAs and four structurally similar saturated ones, in an anaerobic defluorinating enrichment and an activated sludge community. The anaerobic and aerobic transformation/defluorination pathways were elucidated. The results showed that under anaerobic conditions, the α,ß-unsaturation is crucial for FCA biotransformation via reductive defluorination and/or hydrogenation pathways. With sp2 C-F bonds being substituted by C-H bonds, the reductive defluorination became less favorable than hydrogenation. Moreover, for the first time, we reported enhanced degradability and defluorination capability of specific unsaturated FCA structures with trifluoromethyl (-CF3) branches at the α/ß-carbon. Such FCA structures can undergo anaerobic abiotic defluorination in the presence of reducing agents and significant aerobic microbial defluorination. Given the diverse applications and emerging concerns of fluorochemicals, this work not only advances the fundamental understanding of the fate of unsaturated PFASs in natural and engineered environments but also may provide insights into the design of readily degradable fluorinated alternatives to existing PFAS compounds.

The responses of activated sludge to membrane cleaning reagent H2O2 and protection of extracellular polymeric substances.

Hydrogen peroxide (H2O2) is evaluated as a potential replacement for chlorine to control biofouling in membrane bioreactors (MBRs). However, H2O2 might diffuse into the mixed liquor and damage microorganisms during membrane cleaning. This study comprehensively analyzed the impacts of H2O2 on microbes. Key enzymes involved in phenol biodegradation were inhibited with H2O2 concentration increased, and thus phenol degradation efficiency was decreased. Increase of lactic dehydrogenase (LDH) and intracellular reactive oxygen species (ROS) indicated more severe cell rupture with H2O2 concentration increased. At the same H2O2 concentration, Extracellular polymeric substances (EPS) extraction further led to inhibiting the activity of key enzymes, decreasing phenol degradation efficiency, and enhancing LDH release and ROS production, demonstrating that the existence of EPS moderated the adverse impacts on microbes. Spectroscopic characterization revealed the increase of H2O2 decreased tryptophan protein-like substances, protein-associated bonds and polysaccharide-associated bonds. Hydroxyl and amide groups in EPS were attacked, which might lead to the consumption of H2O2, indicated EPS protect the microorganism through sacrificial reaction with H2O2.

Structure-Specific Aerobic Defluorination of Short-Chain Fluorinated Carboxylic Acids by Activated Sludge Communities.

Per- and polyfluoroalkyl substances (PFAS) are a large group of manmade chemicals that impose emerging environmental concerns. Among them, short-chain per- and polyfluorinated carboxylic acids represent an important subgroup used as building blocks of biologically active chemicals and functional materials. Some are also considered PFAS alternatives, and some could be byproducts of the physicochemical treatment of PFAS. However, little is known about the environmental fate of short-chain fluorinated carboxylic acids (FCAs) and their defluorination/transformation by microorganisms. To fill the knowledge gap, we investigated the structure-reactivity relationships in the aerobic defluorination of C3-C5 FCAs by activated sludge communities. Four structures exhibited greater than 20% defluorination, with 3,3,3-trifluoropropionic acid being almost completely defluorinated. We further analyzed the defluorination/transformation pathways and inferred the structures susceptible to aerobic microbial defluorination. We also demonstrated that the defluorination was via cometabolism. The findings advance the fundamental understanding of aerobic microbial defluorination and help assess the environmental fate of PFAS. Since some short-chain PFAS, such as 3,3,3-trifluoropropionic acid, are the incomplete defluorination byproducts of advanced reduction processes, their defluorination by activated sludge communities sheds light on the development of cost-effective chemical-biological PFAS treatment train systems.

Synthetic microbial consortia for biosynthesis and biodegradation: promises and challenges.

Functional differentiation and metabolite exchange enable microbial consortia to perform complex metabolic tasks and efficiently cycle the nutrients. Inspired by the cooperative relationships in environmental microbial consortia, synthetic microbial consortia have great promise for studying the microbial interactions in nature and more importantly for various engineering applications. However, challenges coexist with promises, and the potential of consortium-based technologies is far from being fully harnessed. Thorough understanding of the underlying molecular mechanisms of microbial interactions is greatly needed for the rational design and optimization of defined consortia. These knowledge gaps could be potentially filled with the assistance of the ongoing revolution in systems biology and synthetic biology tools. As current fundamental and technical obstacles down the road being removed, we would expect new avenues with synthetic microbial consortia playing important roles in biological and environmental engineering processes such as bioproduction of desired chemicals and fuels, as well as biodegradation of persistent contaminants.

Topotactic Conversion of Alkali-Treated Intergrown Germanosilicate CIT-13 into Single-Crystalline ECNU-21 Zeolite as Shape-Selective Catalyst for Ethylene Oxide Hydration.

The conversion of the alkali-treated intergrowth germanosilicate CIT-13 into the single-crystalline high-silica ECNU-21 (named after East China Normal University) zeolite, with a novel topology and a highly crystalline zeolite framework, has been realized through a creative top-down strategy involving a mild alkaline-induced multistep process consisting of structural degradation and reconstruction. Instead of acid treatment, hydrolysis in aqueous ammonia solution not only readily cleaved the chemically weak Ge(Si)-O-Ge bonds located within the interlayer double four ring (D4R) units of CIT-13, but also cleaved the metastable Si-O-Si bonds therein. This led to extensive removal of the D4R units, and also generated silanol groups on adjacent silica-rich layers, which then condensed to form a novel daughter structure upon calcination. Individual oxygen bridges in the reassembled ECNU-21 replaced the germanium-rich D4R units in CIT-13, thereby eliminating the original intergrowth phenomenon along the b axis. With an ordered crystalline structure of 10-ring (R) channels as well as suitable germanium-related Lewis acid sites, ECNU-21 serves as a stable solid Lewis acid catalyst for the shape-selective hydration of ethylene oxide (EO) to ethylene glycol (EG) at greatly reduced H2 O/EO ratios and reaction temperature in comparison with the noncatalytic industrial process.

Area-efficient all-digital pulse-shrinking smart temperature sensor with improved accuracy and resolution.

An all-digital CMOS (Complementary Metal-Oxide Semiconductor) pulse-shrinking smart temperature sensor (PSSTS) is proposed to deliver the merits of area efficiency, improved accuracy, and high resolution. First, an inverter-based temperature-sensing delay line generates a pulse with a width proportional to absolute temperature (PTAT). Then, a pulse-shrinking delay line (PSDL) with a pulse-mixing scheme (PMS) measures the PTAT pulse through pulse shrinking. The two delay lines become area efficient when the channel length of transistors is long. Conventionally, the area-efficient PSDL decreases the time resolution. Thus, the all-digital PMS considerably enhances the time resolution and achieves an excellent sensor resolution. Finally, a time subtractor improves sensor accuracy by effectively eliminating the offset error effects. The proposed sensor was implemented in a Taiwan Semiconductor Manufacturing Company 0.35-µm CMOS process and occupied an area of 0.0247 mm2, which is the best area among related studies. The PSSTS exhibited an improved inaccuracy from a maximal inaccuracy of 1.95 °C to 1.5 °C and an excellent resolution of approximately 0.05 °C/LSB. Experimental results prove that this study is area-efficient and achieves improved accuracy and high resolution.

One-step assembly of polymeric demethylcantharate prodrug/Akt1 shRNA complexes for enhanced cancer therapy.

This report demonstrated a one-step assembly for co-delivering chemotherapeutics and therapeutic nucleic acids, constructed by integrating drug molecules into a nucleic acid condensing polymeric prodrug through degradable linkages. Demethylcantharate was selected as the model drug and pre-modified by esterifying its two carboxylic groups with 2-hydroxyethyl acrylate. The synthesized demethylcantharate diacrylate was then used to polymerize with linear polyethyleneimine (PEI 423) through a one-step Michael-addition reaction. The obtained cationic polymeric demethylcantharate prodrug was used to pack Akt1 shRNA into complexes through a one-step assembly. The formed complexes could release the parent drug demethylcantharate and Akt1 shRNA through the hydrolysis of ester bonds. Cellular assays involving cell uptake, cytotoxicity, and cell migration indicated that demethylcantharate and Akt1 shRNA co-delivered in the present form significantly and synergistically suppress the growth and metastasis of three human cancer cells. This work suggests that incorporating drug molecules into a nucleic acid-packing cationic polymer as a polymeric prodrug in a degradable form is a highly convenient and efficient way to co-deliver drugs and nucleic acids for cancer therapy.

Rapid Osseointegration of Titanium Implant With Innovative Nanoporous Surface Modification: Animal Model and Clinical Trial.

OBJECTIVES: SLAffinity is the hybrid topography consisting of micropits and nanoporous TiO2 layers through electrochemical oxidation to mimic the natural bony environment. The aim of this study was to examine the rate of osseointegration in animal models and to further investigate the stability for implants with SLAffinity-treated surface in the clinical trial. MATERIALS AND METHODS: Implants were installed in the mandibular canine-premolar area of 12 miniature pigs. Each pig received 2 implants with the same shapes but with different chemical surfaces. In the clinical trial, 25 patients were included. Each patient received 1 SLAffinity-treated implant on the posterior area of either arch. Resonance frequency analysis and computed tomography were assessed weekly over the first 12 weeks after implant placement. RESULTS: The results found that surface treatment did affect the bone-to-implant contact (BIC) significantly. Comparison of BIC at 3 weeks in animal study showed that the SLAffinity-treated implants presented significantly higher values than machine surface implants. SLAffinity-treated implants also proved clinically successful through 12 months, ready for prosthodontic restoration. CONCLUSION: The effect of SLAffinity treatments enhanced osseointegration significantly, especially at early stages of bone healing. Clinical trial finding, furthermore, ensured that the SLAffinity treatment was a reliable surface modification alternative.

Effects of sandblasting media and steam cleaning on bond strength of titanium-porcelain.

The effects of sandblasting media and steam cleaning treatment after sandblast were examined on tensile bond strength of porcelain to titanium. The use of the commercially available silica-coated alumina particles for sandblast was significantly effective for increasing bond strength than the conventional alumina. It might be due to the increased surface roughness and existence of remaining silica on titanium surface. Additional application of the steam cleaning on titanium surface after sandblasting could make the surface configuration clear in SEM by removing some sandblasted particles loosely embedded in titanium as well as the debris and oily contaminants. The resultant bond strength was significantly improved to reach almost the maximum strength of this porcelain-titanium system regardless of the kind of sandblasting media used, which was confirmed by the observation of the failure mode showing that most of the fracture surface was occupied by cohesive failure in porcelain.

A brief report on the normal range of forehead temperature as determined by noncontact, handheld, infrared thermometer.

BACKGROUND: Noncontact forehead temperature measurement by handheld infrared thermometer was used as a screening tool for fever. However, the accuracy data and normal range of forehead temperature determined by this method were not available. METHODS: The temperature readings from 3 handheld infrared thermometers were validated against an electronic thermometer. Normal range of forehead temperature was determined by measuring the forehead temperature in 1000 apparently healthy subjects. RESULTS: Significant differences were detected in readings obtained by the 3 different handheld infrared thermometers (analysis of covariance, P < .001) The most accurate one was chosen, and the normal range of forehead temperature in 1000 subjects detected by this method was 31.0 degrees C to 35.6 degrees C. CONCLUSIONS: Our study shows that commercially available, handheld infrared thermometers require individual validation. Forehead temperature in excess of 35.6 degrees C is suggestive of fever. Further studies are required to confirm accuracy of this value in detecting fever.

DNA-based subtyping of glycogen storage disease type III: mutation and haplotype analysis of the AGL gene in Chinese.

Glycogen storage disease type III (GSD III) is an inborn error of glycogen metabolism caused by a deficiency of glycogen debranching enzyme (AGL). Here, we investigate two unrelated Hong Kong Chinese GSD III patients and identify a novel 5-base pair deletional mutation, 2715_2719delTCAGAin exon 22, in one patient and a nonsense mutation, 1222C>T (R408X) in exon 11, in another patient. Since GSD IIIb is only caused by mutation in exon 3 of the AGL gene, we diagnose our patients to have GSD IIIa, which is consistent with the clinical diagnosis. Until now, R408X has only been reported in Faroe Islands GSDIII patients and was thought to demonstrate a founder effect. In this study, haplotyping of the disease-bearing chromosomes in the AGL locus by 19 intragenic single nucleotide polymorphisms shows that R408X is linked with IVS16+8T and IVS23-21T in our patient while R408X is linked with IVS16+8C and IVS23-21A in the Faroe Islands. The different haplotypes of R408X in Chinese and Faroese indicated that R408X is a recurrent mutation.