Gas-permeable Membrane Research Articles

Modeling nitrogen recovery and water transport in gas-permeable membranes

This study presents a new modeling approach for nitrogen recovery in gas-permeable membrane (GPM) contactors, including both ammonia and water transport dynamics. A distinct feature of the model is its capacity to model water transport across the membrane, which has been overlooked in most publications. Osmotic pressure differences are used to predict the behavior of ammonia and water transport in the GPM contactor. Experiments carried out to develop, test and calibrate the model examined the dynamics of ammonia and water transport through the GPM contactor at various nitrogen concentrations. Specifically, the GPM contactor was tested for nitrogen recovery from high-strength synthetic wastewaters (2.4–10.6 g N/L) at 35 °C and at pH 9. The initial volume of the trapping solution (diluted H2SO4) was 10 times lower than that of the synthetic wastewater, aiming to concentrate the recovered nitrogen. The estimated ammonia transport constant (Km) ranged between (1.2 - 2.1)·10–6 m/s and water transport constant Kw between (2.8 - 8.2)·10–10 m/(s bar). Numerical determination of the model parameters revealed high R² values, demonstrating strong agreement with experimental data.

Optimizing biomass production and photochemical efficiency of in vitro-grown Capsicum annuum L. using gas-permeable membrane flasks

Optimizing biomass production and photochemical efficiency of in vitro-grown Capsicum annuum L. using gas-permeable membrane flasks

Production of Volatile Fatty Acids from Cheese Whey and Their Recovery Using Gas-Permeable Membranes

The use of anaerobic fermentation to produce volatile fatty acids (VFAs) is an environmentally sustainable alternative for cheese whey (CW) valorization. This study evaluates the effect of pH control on the conversion of organic matter to VFAs from CW and assesses VFA recovery using a novel approach based on gas-permeable membranes. VFA bioconversion and composition were studied with initial and sequential control of pH, both in acidic and alkaline conditions. Bioconversion efficiencies for assays with initial pH control were 36% and 45% for acidic and alkaline conditions, respectively. Sequential control of pH resulted in an increase in bioconversion to 54% under acidic conditions. Under acidic conditions, a variety of VFA was produced (mainly butyric, acetic, and propionic acids), while under alkaline conditions the majority was acetic acid. VFA recovery using a novel system of tubular gas-permeable membranes accounted for 15% and 100% of the total VFA from effluent 1 (butyric, acetic, and propionic acids) and effluent 2 (mainly acetic acid), respectively.

Scalable process development of NK and CAR-NK expansion in a closed bioreactor.

Production of large amounts of functional NK and CAR-NK cells represents one of the bottlenecks for NK-based immunotherapy. In this study, we developed a large-scale, reliable, and practicable NK and CAR-NK production using G-Rex 100M bioreactors, which depend on a gas-permeable membrane technology. This system holds large volumes of medium with enhanced oxygen delivery, creating conditions conducive to large-scale PBNK and CAR-NK expansions for cancer therapy. Both peripheral blood NK cells (PBNKs) and CAR-NKs expanded in these bioreactors retained similar immunophenotypes and exhibited comparable cytotoxicity towards hepatocellular carcinoma (HCC) cells akin to that of NK and CAR-NK cells expanded in G-Rex 6 well bioreactors. Importantly, cryopreservation minimally affected the cytotoxicity of NK cells expanded using the G-Rex 100M bioreactors, establishing a robust platform for scaled-up NK and CAR-NK cell production. This method is promising for the development of "off-the-shelf" NK cells, supporting the future clinical implementation of NK cell immunotherapy.

Thermal Vapor Deposition of a Hydrophobic and Gas-Permeable Membrane on Zirconium Phosphate Cation Exchanger: An Oral Sorbent for the Urea Removal of Kidney Failure.

An oral sorbent with high capacity for NH4+ is desirable in lowering the blood urea level and mitigating the dialysis burden for end-stage kidney disease (ESKD) patients. Zirconium phosphate (ZrP) is an amorphous cation ion exchanger with high NH4+ binding capacity as a sorbent material, but its selectivity to remove NH4+ is limited in the presence of other competing ions in water solution. We previously have developed a gas-permeable and hydrophobic perfluorocarbon coating on ZrP, which improves ZrP's NH4+ selectivity. However, the coating preparation procedure, a wet chemistry approach, is complicated and time-consuming, and more importantly, the large amount of usage of acetone poses a concern for the application of ZrP as an oral sorbent. In this study, we developed a solventless coating protocol that effectively coats ZrP with tetraethyl orthosilicate (TEOS) and 1H,1H,2H,2H-perfluorooctyltriethoxysilane (FOTS) via thermal vapor deposition (TVD) in a simplified manner. X-ray photoelectron spectroscopy (XPS) and contact angle measurements verify the two coatings are successfully deposited on the ZrP surface, and the coating condition was optimized based on an in vitro static binding study. The dynamic binding study of competing ions on Na-loaded ZrP with TVD coatings yields a maximum NH4+ removal (∼3.2 mequiv/g), which can be improved to ∼4.7 mequiv/g if H-loaded ZrP under the same coating condition is used in basic stock solutions. More importantly, both materials barely remove Ca2+ and show excellent acid resistance. The significant improvement in the NH4+ binding capacity and selectivity reported here establishes a highly promising surface modification approach to optimize oral sorbents for ESKD patients.

Gas permeable membrane electrode assembly with in situ utilization of authigenic acid and base for transmembrane electro-chemisorption to enhance ammonia recovery from wastewater

Ammonia recovery from wastewater is of great significance for aquatic ecology safety, human health and carbon emissions reduction. Electrochemical methods have gained increasing attention since the authigenic base and acid of electrochemical systems can be used as stripper and absorbent for transmembrane chemisorption of ammonia, respectively. However, the separation of electrodes and gas permeable membrane (GPM) significantly restricts the ammonia transfer-transformation process and the authigenic acid-base utilization. To break the restrictions, this study developed a gas permeable membrane electrode assembly (GPMEA), which innovatively integrated anode and cathode on each side of GPM through easy phase inversion of polyvinylidene fluoride binder, respectively. With the GPMEA assembled in a stacked transmembrane electro-chemisorption (sTMECS) system, in situ utilization of authigenic acid and base for transmembrane electro-chemisorption of ammonia was achieved to enhance the ammonia recovery from wastewater. At current density of 60 A/m2, the transmembrane ammonia flux of the GPMEA was 693.0 ± 15.0 g N/(m2·d), which was 86 % and 28 % higher than those of separate GPM and membrane cathode, respectively. The specific energy consumption of the GPMEA was 9.7∼16.1 kWh/kg N, which were about 50 % and 25 % lower than that of separate GPM and membrane cathode, respectively. Moreover, the application of GPMEA in the ammonia recovery from wastewater is easy to scale up in the sTMECS system. Accordingly, with the features of excellent performance, energy saving and easy scale-up, the GPMEA showed good prospects in electrochemical ammonia recovery from wastewater.

Mathematical model and electronic system for real-time O2 control in storage boxes: Development and validation under fluctuating temperatures

Modified atmosphere storage containers with gas-permeable membranes are effective in reducing post-harvest losses, mainly at low temperatures but become ineffective in controlling O2 and CO2 at temperatures above 3 °C. This study aimed to develop a method to control O2 in a storage box. This method uses a diffusion tube, that blocks air from entering the box but allows for air exchange when an air blower is activated. The blower ON frequency (BOF) was modelled as a function of temperature, considering the type and amount of produce, blower properties, tube dimension, and the O2 setpoint. The effects of temperature and gas composition on respiration rate kinetics were analysed with Michaelis-Menten and Arrhenius equations. The model was used to program the microcontroller to control the blower. Validation was performed using a 190 L box containing 25 kg of broccoli. The developed model maintained the desired O2 concentration under fluctuating temperatures. The BOF values ranged from 25.6 to 549.2 s h−1, for temperature fluctuations between 4 and 23 °C. The time required to reach the 3% O2 in the box differed depending on the blower's state. With the blower turned ON from the beginning, it took 24.9 h. However, when the blower was initially turned OFF, it took 11.1 h. Despite temperature changes, the system maintained O2 at 3.8 ± 0.29% and CO2 at 14.4 ± 0.66%. The system is promising for commercial use and best suited for CO2 tolerant produce because it requires a separate mathematical model and control mechanism.

Development of an autonomous on-site dissolved inorganic carbon analyzer using conductometric detection

BackgroundThe increase in anthropogenic CO2 concentrations in the Earth's atmosphere since the industrial revolution has resulted in an increased uptake of CO2 by the oceans, leading to ocean acidification. Dissolved Inorganic Carbon (DIC) is one of the key variables to characterize the seawater carbonate system. High quality DIC observations at a high spatial-temporal resolution is required to improve our understanding of the marine carbonate system. To meet the requirements, autonomous DIC analyzers are needed which offer a high sampling frequency, are cost-effective and have a low reagent and power consumption. ResultsWe present the development and validation of a novel analyzer for autonomous measurements of DIC in seawater using conductometric detection. The analyzer employs a gas diffusion sequential injection approach in a “Tube In A Tube” configuration that facilitates diffusion of gaseous CO2 from an acidified sample through a gas permeable membrane into a stream of an alkaline solution. The change in conductivity in the alkaline medium is proportional to the DIC concentration of the sample and is measured using a detection cell constructed of 4 hollow brass electrodes. Physical and chemical optimizations of the analyzer yielded a sampling frequency of 4 samples h−1 using sub mL reagent volumes for each measurement. Temperature and salinity effects on DIC measurements were mathematically corrected to increase accuracy. Analytical precision of ±4.9 μmol kg−1 and ±9.7 μmol kg−1 were achieved from measurements of a DIC reference material in the laboratory and during a field deployment in the southwest Baltic Sea, respectively. SignificanceThis study describes a simple, cost-effective, autonomous, on-site benchtop DIC analyzer capable of measuring DIC in seawater at a high temporal resolution as a step towards an underwater DIC sensor. The analyzer is able to measure a wide range of DIC concentrations in both fresh and marine waters. The achieved accuracy and precision offer an excellent opportunity to employ the analyzer for ocean acidification studies and CO2 leakage detection in the context of Carbon Capture and Storage operations.

Bubble Turbulent Gas-Permeable Membrane for Ammonia Recovery from Swine Wastewater: Mass Transfer Enhancement and Antifouling Mechanisms.

Recovering ammonium from swine wastewater employing a gas-permeable membrane (GM) has potential but suffers from the limitations of unattractive mass transfer and poor-tolerance antifouling properties. Turbulence is an effective approach to enhancing the release of volatile ammonia from wastewater while relying on interfacial disturbance to interfere with contaminant adhesion. Herein, we design an innovative gas-permeable membrane coupled with bubble turbulence (BT-GM) that enhances mass transfer while mitigating membrane fouling. Bubbles act as turbulence carriers to accelerate the release and migration of ammonia from the liquid phase, increasing the ammonia concentration gradient at the membrane-liquid interface. In comparison, the ammonium mass transfer rate of the BT-GM process applied to real swine wastewater is 38% higher than that of conventional GM (12 h). Through a computational fluid dynamics simulation, the turbulence kinetic energy of BT-GM system is 3 orders of magnitude higher than that of GM, and the effective mass transfer area is nearly 3 times that of GM. Seven batches of tests confirmed that the BT-GM system exhibits remarkable antifouling ability, broadens its adaptability to complex water quality, and practically promotes the development of sustainable resource recycling.

Efficient sulfide and methane removal in anaerobic secondary effluent using a pilot-scale membrane-aerated biofilm reactor

Anaerobic secondary treatment can enable energy-efficient removal of organic matter but may produce effluent containing dissolved methane and sulfide that must be managed before discharge or reuse. In this study, we operated a Membrane-aerated Biofilm Reactor (MABR) to achieve reliable removal of dissolved sulfide and methane from anaerobic secondary effluent at pilot-scale. The pilot-scale MABR was equipped with gas permeable polymethyl pentene (PMP) membranes, promoting surface growth of aerobic biofilm via diffusion-based aeration (lumen-to-surface diffusion). The system treated anaerobic secondary effluent from a demonstration-scale anaerobic membrane bioreactor (AnMBR) processing 90 m3/d primary effluent. MABR influent flow rate was increased from 8.2 to 32.7 m3/d to elevate substrate loading rates to the biofilm. The MABR consistently achieved >99 % removal of sulfide and dissolved methane, even at the maximum substrate loading rate: 2.3 g-S/m2/d for sulfide and 2.5 g-CH4/m2/d for dissolved methane. Despite effective sulfide and methane removal, incomplete nitrification (<25 % ammonia removal) occurred, with a portion of ammonia converted into nitrous oxide (N2O), a greenhouse gas 298 times more potent than CO2. Operating the MABR incurred low energy costs: 0.01 to 0.05 kWh/m3 for the compressor supplying air to the membrane lumen and 0.01 kWh/m3 for the influent pump. An in-depth mass balance of N2O emissions from the MABR revealed that N2O was subject to counter-diffusion (surface-to-lumen diffusion) in which >99 % of the N2O produced within the biofilm was recovered in off-gas from the gas permeable membranes (hollow fibers).

Combined leaf gas-exchange system for model assessment.

Leaf gas-exchange measurements are useful in assessing plant environmental responses. However, uncertainties in the leaf gas-exchange model potentially limit its application. The main challenge in the model-dependent calculations is to detect violations of assumptions. Here, we developed a system that integrates into one instrument the direct measurement of leaf intercellular CO2 concentration and the standard open-flow (OF) and novel open-diffusion (OD) systems for flux measurement. In the OD system, a gas-permeable membrane between the leaf ambient air and outside air creates CO2 and H2O differentials, rather than the air flow in the OF chamber. We measured hypostomatous and amphistomatous leaves of several species with different photosynthetic capacities [sunflower (Helianthus annuus), grape (Vitis vinifera), lemon (Citrus limon), and cherry (Prunus avium)]. The CO2 and H2O differentials in the OD system strictly depend on the flux measured by the OF system. The lower permeability of the membrane resulted in a larger differential per flux, indicating that the OD system can increase the resolution for a small flux. An analysis of the conductance model along with observations suggested that cuticle and leaf intercellular conductances and the unsaturation of leaf humidity contributed to discrepancies between the direct measurement and standard calculation. The combined system developed here provides an opportunity to address these overlooked concepts in leaf gas exchange.

Acidogenic fermentation of biowaste coupled with nitrogen recovery using selective membranes to produce a VFA-rich liquid with a high C/N ratio

This research focuses on the production of a liquid stream rich in volatile fatty acids (VFAs) and low ammoniacal nitrogen content (<0.1 g N/L) from biowaste. The liquid stream was obtained by combining (i) mixed culture acidogenic fermentation to maximise VFA production and (ii) gas-permeable membrane (GPM) contactor to recover ammoniacal nitrogen. Three batch fermentation tests of biowaste collected in a full-scale mechanical-biological treatment plant provided high and stable VFA concentrations (37–39 g CODVFA/L). VFAs represented 73–81 % of the soluble chemical oxygen demand (sCOD) concentration, with a predominance of acetic, propionic and butyric acids. A highly specialized microbial community was observed in all batch tests, with Bacteroidota and Firmicutes as predominant phyla (>90 % of relative abundance). The GPM contactor recovered more than 99 % of the ammoniacal nitrogen in the fermentation liquid without VFA losses. The suitability of the produced fermentation liquid with a high C/N ratio for downstream applications was evaluated using biomethane potential tests (BMP) at different total ammonium nitrogen (TAN) concentrations (0.76–3.15 g N/L) and circumneutral pH. Despite achieving similar ultimate methane yields (279–314 NmL CH4/g CODfeed), lower TAN concentrations in the biowaste fermentation liquid improved anaerobic biodegradation kinetics, enhancing its potential applicability for methane production.

Ammoniacal nitrogen recovery from swine slurry using a gas-permeable membrane: pH control strategies and feed-to-trapping volume ratio.

Gas-permeable membrane (GPM) technology is gaining interest to recover nitrogen from residual effluents due to its effectiveness, simple operation and capacity of producing a nutrient rich product with fertilising value. In this study, a GPM contactor was used at 25°C to recover total ammoniacal nitrogen (TAN) from swine slurry as a concentrated (NH4)2SO4 solution. Firstly, a synthetic solution was tested on a wide pH range (6-12). Results showed that the ammonia mass transfer constants (Km) increased from 7.9·10-9 to 1.2·10-6m/s as the pH increased. The reagent consumption to control the pH per mole nitrogen recovered had a minimum at pH 9, which showed a Km value of 3.0·10-7m/s. Secondly, various pH control strategies were tested using swine slurry, including (i) no pH control, (ii) pH control at 8.5, 9.0 and 10.0, and (iii) an initial spike of the NaOH equivalent to the required to control the pH at 9. The test without pH control reached a TAN recovery of around 60%, which could be an interesting strategy when high nitrogen recoveries or short operating times are not required. The pH control at 9 stood out as the most favourable operating condition due to its high Km and lower reagent consumption. Thirdly, several feed-to-trapping volume ratios ranging from 1:1 to 15:1 were tested using swine slurry at pH 9. These assays revealed that a GPM process with a high feed-to-trapping volume ratio fastens the recovery of 99% of TAN as a high purity (NH4)2SO4 solution containing 40g N/L.

Enhanced ammonia recovery from wastewater by a transmembrane electro-chemisorption system directly connecting anode chamber and cathode chamber with gas permeable membrane

A transmembrane electro-chemisorption (TMECS) system was developed via directly connecting anode chamber (AnC) and cathode chamber (CaC) with a hydrophobic gas permeable membrane (GPM). The special structure of the TMECS system with AnC and CaC at two sides of GPM contributed to both forming localized extreme pH layers at two sides of GPM and using electrical field forces to enrich NH4+, thus realizing on-site utilization of authigenic acid and base to enhance the ammonia recovery from wastewater. Increasing applied voltage (from 2.0 to 6.0 V) and salt concentration (from 80 to 500 mM) could improve the system performance by promoting water splitting and reducing internal resistance, respectively. The system showed wide applicability for the influent ammonia concentration (from 560 to 5000 mg NH4+-N/L) with the ammonia removal and recovery efficiencies of 61.9 %–77.1 % and 78.5 %–91.9 % at 24 h, respectively. For high influent ammonia concentration (5000 mg NH4+-N/L), a specific energy consumption of 21.8 kWh/kg N could be reached. Moreover, the system can be easily scaled up by stacking the TMECS units. Therefore, the system shows good prospects in saving chemicals, wide applicability and easy scale-up for ammonia recovery from wastewater.

New Trend of Amperometric Gas Sensors Using Atomic Gold-Decorated Platinum/Polyaniline Composites

The Amperometric Gas Sensor (AGS) uses an electrode as the transducer element which converts its signal into a current from the electrochemical reaction of analytes taking place at the electrode surface. Many attempts to improve AGS performance, such as modifying the working electrode, applying a particular gas-permeable membrane, and selecting the proper electrolyte, etc., have been reported in the scientific literature. On the other hand, in the materials community, atomic gold has gained much attention because its physicochemical properties dramatically differ from those of gold nanoparticles. This paper provides an overview of the use of atomic gold in AGSs, both in a bulky AGS and a miniaturized AGS. In the miniaturized AGS, the system must be redesigned; for example, the aqueous electrolyte commonly used in a bulky AGS cannot be used due to volatility and fluidity issues. A Room Temperature Ionic Liquid (RTIL) can be used to replace the aqueous electrolyte since it has negligible vapor pressure; thus, a thin film of RTIL can be realized in a miniaturized AGS. In this paper, we also explain the possibility of using RTIL for a miniaturized AGS by incorporating a quartz crystal microbalance sensor. Several RTILs coated onto modified electrodes used for isomeric gas measurement are presented. Based on the results, the bulky and miniaturized AGS with atomic gold exhibited a higher sensor response than the AGS without atomic gold.

Ammonia recovery from landfill leachate using gas-permeable membranes: Comparison of membrane fibers and expanded membrane tubes in both in-situ and shell-tube configurations

This study rigorously assessed the efficacy of gas-permeable membrane (GPM) technologies in recovering ammonia from landfill leachate, focusing on polypropylene (PP) and expanded polytetrafluoroethylene (ePTFE) membranes within in-situ (INS) and shell-tube (ST) configurations. The research spanned an array of determinants, including influencing factors, sustained operational efficiency, fouling dynamics, and recovery product purity. It was established that the INS-ePTFE configuration demonstrated formidable recovery capabilities and durable performance after fouling cleaning, with ammonia removal efficiency resurging to its initial state and stabilizing at 93 % post-maintenance. In contrast, the ST-PP configuration experienced an unrecoverable drop in removal efficiency to 83 %. Fouling analysis indicated that ePTFE membranes retained structural stability against organic and inorganic deposition, with humic substances and metal carbonates as predominant constituents. Conversely, PP membranes exhibited a decline in hydrophobic properties and structural integrity upon contamination. Furthermore, pH elevation significantly enhanced ammonia nitrogen mass transfer coefficients without increasing osmotic water transfer in INS systems. The INS system also showed promise for synergistic phosphorus recovery within the pH of 9 to 10. Then, NO3–, SO42- and humic acid did not impede ammonia translocation, whereas a 1.5 % mass fraction of Ca2+ incurred a roughly 30 % reduction in ammonia mass transfer coefficients across each system. The recovery of high purity (NH4)2SO4 attested to the exceptional retention capabilities of GPMs for organics and ions, with rejection rates surpassing 99.6 %. The comprehensive analysis underscored the operational superiority of the INS-ePTFE configuration in ammonia recovery.

Gas equilibrium membrane inlet mass spectrometry (GE-MIMS) for water at high pressure

Abstract. Gas species are widely used as natural or artificial tracers to study fluid dynamics in environmental and geological systems. The recently developed gas equilibrium membrane inlet mass spectrometry (GE-MIMS) method is most useful for accurate and autonomous on-site quantification of dissolved gases in aquatic systems. GE-MIMS works by pumping water through a gas equilibrator module containing a gas headspace, which is separated from the water by a gas-permeable membrane. The partial pressures of the gas species in the headspace equilibrate with the gas concentrations in the water according to Henry's Law and are quantified with a mass spectrometer optimized for low gas consumption (miniRUEDI or similar). However, the fragile membrane structures of the commonly used equilibrator modules break down at water pressures ≳3 bar. These modules are therefore not suitable for use in deep geological systems or other environments with high water pressures. To this end, the SysMoG® MD membrane module (Solexperts AG, Switzerland; “SOMM”) was developed to withstand water pressures of up to 100 bar. Compared to the conventionally used GE-MIMS equilibrator modules, the mechanically robust construction of the SOMM module entails slow and potentially incomplete gas–water equilibration. We tested the gas equilibration efficiency of the SOMM and developed an adapted protocol that allows correct operation of the SOMM for GE-MIMS analysis at high water pressures. This adapted SOMM GE-MIMS technique exhibits a very low gas consumption from the SOMM to maintain the gas–water equilibrium according to Henry's Law and provides the same analytical accuracy and precision as the conventional GE-MIMS technique. The analytical potential of the adapted SOMM GE-MIMS technique was demonstrated in a high-pressure fluid migration experiment in an underground rock laboratory. The new technique overcomes the pressure limitations of conventional gas equilibrators and thereby opens new opportunities for efficient and autonomous on-site quantification of dissolved gases in high-pressure environments, such as in research and monitoring of underground storage of CO2 and waste deposits or in the exploration of natural resources.

Therapeutic Effects of Aβ-Specific Regulatory T Cells in Alzheimer's Disease: A Study in 5xFAD Mice.

The aging global population is placing an increasing burden on healthcare systems, and the social impact of Alzheimer's disease (AD) is on the rise. However, the availability of safe and effective treatments for AD remains limited. Adoptive Treg therapy has been explored for treating neurodegenerative diseases, including AD. To facilitate the clinical application of Treg therapy, we developed a Treg preparation protocol and highlighted the therapeutic effects of Tregs in 5xFAD mice. CD4+CD25+ Tregs, isolated after Aβ stimulation and expanded using a G-rex plate with a gas-permeable membrane, were adoptively transferred into 5xFAD mice. Behavioral analysis was conducted using Y-maze and passive avoidance tests. Additionally, we measured levels of Aβ, phosphorylated tau (pTAU), and nitric oxide synthase 2 (NOS2) in the hippocampus. Real-time RT-PCR was employed to assess the mRNA levels of pro- and anti-inflammatory markers. Our findings indicate that Aβ-specific Tregs not only improved cognitive function but also reduced Aβ and pTAU accumulation in the hippocampus of 5xFAD mice. They also inhibited microglial neuroinflammation. These effects were observed at doses as low as 1.5 × 103 cells/head. Collectively, our results demonstrate that Aβ-specific Tregs can mitigate AD pathology in 5xFAD mice.

Silicone rubber membrane devices permit islet culture at high density without adverse effects.

Conventional culture conditions, such as in T-flasks, require that oxygen diffuse through the medium to reach the islets; in turn, islet surface area density is limited by oxygen availability. To culture a typical clinical islet preparation may require more than 20 T-175 flasks at the standard surface area density of 200IE/cm2. To circumvent this logistical constraint, we tested islets cultured on top of silicon gas-permeable (GP) membranes which place islets in close proximity to ambient oxygen. Oxygenation of individual islets under three culture conditions, standard low-density, non-GP high density, and GP high density, were first modeled with finite element simulations. Porcine islets from 30 preparations were cultured for 2days in devices with GP membrane bottoms or in paired cultures under conventional conditions. Islets were seeded at high density (HD, ∼4000IE/cm2, as measured by DNA) in both GP and non-GP devices. In simulations, individual islets under standard culture conditions and high density cultures on GP membranes were both well oxygenated whereas non-GP high density cultured islets were anoxic. Similarly, compared to the non-GP paired controls, islet viability and recovery were significantly increased in HD GP cultures. The diabetes reversal rate in nude diabetic mice was similar for HD GP devices and standard cultures but was minimal with non-GP HD cultures. Culturing islets in GP devices allows for a 20-fold increase of islet surface area density, greatly simplifying the culture process while maintaining islet viability and metabolism.

Quest for Nitrous Oxide-reducing Bacteria Present in an Anammox Biofilm Fed with Nitrous Oxide.

N2O-reducing bacteria have been examined and harnessed to develop technologies that reduce the emission of N2O, a greenhouse gas produced by biological nitrogen removal. Recent investigations using omics and physiological activity approaches have revealed the ecophysiologies of these bacteria during nitrogen removal. Nevertheless, their involvement in‍ ‍anammox processes remain unclear. Therefore, the present study investigated the identity, genetic potential, and activity‍ ‍of N2O reducers in an anammox reactor. We hypothesized that N2O is limiting for N2O-reducing bacteria‍ ‍and an‍ ‍exogeneous N2O supply enriches as-yet-uncultured N2O-reducing bacteria. We conducted a 1200-day incubation of N2O-reducing bacteria in an anammox consortium using gas-permeable membrane biofilm reactors (MBfRs), which efficiently supply N2O in a bubbleless form directly to a biofilm grown on a gas-permeable membrane. A 15N tracer test indicated that the supply of N2O resulted in an enriched biomass with a higher N2O sink potential. Quantitative PCR and 16S rRNA amplicon sequencing revealed Clade II nosZ type-carrying N2O-reducing bacteria as protagonists of N2O sinks. Shotgun metagenomics showed the genetic potentials of the predominant Clade II nosZ-carrying bacteria, Anaerolineae and Ignavibacteria in MBfRs. Gemmatimonadota and non-anammox Planctomycetota increased their abundance in MBfRs despite their overall lower abundance. The implication of N2O as an inhibitory compound scavenging vitamin B12, which is essential for the synthesis of methionine, suggested its limited suppressive effect on the growth of B12-dependent bacteria, including N2O reducers. We identified Dehalococcoidia and Clostridia as predominant N2O sinks in an anammox consortium fed exogenous N2O because of the higher metabolic potential of vitamin B12-dependent biosynthesis.