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The study analyses the performance of a pilot plant using a rotating hollow fibre (HF) membrane bioreactor system. The experiments evaluated the effect of operational parameters such as rotational speed, aeration strategies, and maintenance cleaning (MC) procedures on the efficiency of the system, in particular transmembrane pressure (TMP) and filtrate quality. The results indicate that the rotating membrane module reduces TMP increase and can operate for 48 days with satisfactory performance, even without aeration. This has the potential to significantly improve efficiency, resulting in significant energy savings. In addition, two MC methods, clean in air and clean in place, were tested and found to be efficient for weekly MC. It was observed that operating without aeration during colder seasons may not be effective. Therefore, adaptive strategies are needed to address seasonal temperature variations.

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OBJECTIVE: Carbapenem-resistant Acinetobacter baumannii (CRAB) is a global concern as effective treatments are very limited. We previously used a modified susceptibility testing approach to predict growth suppression in carbapenem-resistant Enterobacterales, but there are uncertainties about the generalizability of the model. The objective of this study is to verify if a similar approach can be extended to CRAB. METHOD: A clinical isolate of CRAB resistant to ceftazidime/avibactam (CAZ/AVI, MIC = 32/4 mg/L) was examined. CAZ susceptibility was determined using increasing concentrations of AVI (0-64 mg/L), and MIC reduction was characterized with a sigmoid inhibitory maximum effect (Emax) model. The effectiveness of CAZ/AVI was validated in a hollow fibre infection model (HFIM) over 72 hours, using simulated unbound serum / epithelial lining fluid (ELF) exposures of 2.5 g over 2 hours every 8 hours. Baseline inocula of approximately 5.5 log CFU/mL were examined. RESULTS: An AVI concentration-dependent reduction in CAZ MIC was observed (r2 = 0.99). CAZ MIC was dramatically reduced from 512 mg/L (no AVI) to 32 mg/L (AVI = 4 mg/L), and further to 8 mg/L (AVI = 16 mg/L). Pharmacokinetic simulations were satisfactory in the HFIM (r2 > 0.96). Bacterial suppression was observed >24 hours with the serum exposure, but not that from the ELF. CONCLUSION: Using multiple AVI concentrations within the clinically relevant range, our susceptibility testing approach could have better insights of treatment outcome for infections caused by CRAB. This could potentially lead to effective intervention(s) overlooked by conventional susceptibility testing method. This case highlights the importance of site-specific drug exposures on determining treatment outcome.

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OBJECTIVES: The proliferation of metallo-ß-lactamase (MBL)-producing Pseudomonas aeruginosa represents a significant public health threat. P. aeruginosa undergoes significant phenotypic changes that drastically impair antibiotic efficacy. The objectives of this study were (1) to quantify the time-course of killing of VIM-2-producing P. aeruginosa in response to aztreonam-based therapies (including avibactam for coverage of AmpC), and (2) to document the capacity of P. aeruginosa to undergo morphological transformations that facilitate persistence. METHODS: A well-characterised, clinical VIM-2-producing P. aeruginosa was studied in the hollow fibre infection model (HFIM) over 9 days (7 days of active antibiotic therapy, 2 days of treatment withdrawal) at a 107.5 CFU/mL starting inoculum. HFIM treatment arms included: growth control, aztreonam, ceftazidime/avibactam, aztreonam/ceftazidime/avibactam, polymyxin B, and aztreonam/ceftazidime/avibactam/polymyxin B. In addition, real-time imaging studies were conducted under static conditions to determine the time course of the reversion of persister cells. RESULTS: There was a pronounced discrepancy between OD620 and bacterial counts obtained from plating methods (hereafter referred to as 'OD-count discrepancy'). For aztreonam monotherapy, observed counts were 0 CFU/mL by 120 h. Despite this, there was a significant OD-count discrepancy compared with the pre-treatment 0 h. Between therapy withdrawal at 168 h and 216 h, all arms with suppressed counts had regrown to the system-carrying capacity. Real-time imaging of the P. aeruginosa filaments after drug removal showed rapid reversion from a long, filamentous phenotype to many individual rods within 2 h. CONCLUSION: Managing MBL-producing P. aeruginosa requires a multifaceted approach, focused on maximising killing and minimising proliferation of resistant and persistent subpopulations, which will involve eliminating drug-induced phenotypic transformers.

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A multifunctional hollow fibre was prepared by the modification of polyethersulfone (PES) with laccase (Lac) and phosphorus-doped graphitic carbon nitride (P-gC3N4) for the removal of ciprofloxacin and sulfamethoxazole. The properties and structure elucidation of the prepared membranes were evaluated using contact angle analysis, Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), correlative light electron microscopy (CLEM), atomic force microscopy (AFM), tensile strength, water-intake capacity, and pure water flux. The modified multifunctional hollow fibre membranes showed increased root mean square surface roughness from 50 nm for neat PES to 104 nm, which contributed to the significantly higher water flux of 90 L.m-2h-1 compared to 54 L.m-2h-1 for pristine PES. The hydrophilicity also improved after modification as the contact angle reduced from 72° ± 1.01° to 42° ± 2.26°. The modified hollow fibre membranes showed an enhanced removal of ciprofloxacin (77%) and sulfamethoxazole (80%). Moreover, antifouling properties towards bovine serum albumin were 89% for FRR, 7% for Rr, 9% for Rir and 17% for Rt. Regeneration studies showed that the multifunctional hollow fibre membrane obtained a high removal percentage of 79% towards sulfamethoxazole after five cycles. Hence, this work proposes a new system that can be successfully utilized in the treatment of emerging pharmaceutical pollutants in water.

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INTRODUCTION: Guideline-based therapy for Mycobacterium avium complex (MAC) pulmonary disease achieves sustained sputum conversion rates in only 43-53% of patients. Repurposing of ß-lactam antibiotics such as ertapenem could expedite design of more efficacious regimens, compared to developing new drugs. METHODS: We performed an ertapenem exposure-response study in the hollow fibre system model of intracellular MAC (HFS-MAC). We recapitulated human-like intrapulmonary concentration-time profiles of eight once-daily intravenous doses of ertapenem over 28 days and performed repetitive sampling for drug concentration-time profiles and MAC burden. The % of time concentration persisted above MIC (%TMIC) mediating either 50% or 80% of maximal effect (E50, EC80) were identified. The EC80 was used as target exposure in a 10 000 subject Monte Carlo experiments for ertapenem doses of 1G, 2G, or 4G administered once versus twice daily. RESULTS: The ertapenem MIC ranged from 0.5 to 2 mg/L on three occasions. Ertapenem achieved a half-life of 4.04 ± 0.80 h in the HFS-MAC and killed a maximum of 2.17 log10 CFU/mL below day 0. The EC50 was %TMIC of 75.9% (95% confidence interval: 68.43%-86.54%) and the EC80 was %TMIC of 100%. Target attainment probability was >90% for 1G twice daily up to an MIC of 2 mg/L, while for 2G twice daily the susceptibility MIC breakpoint was 4-8 mg/L. CONCLUSIONS: Ertapenem microbial kill below day 0 burden was better than guideline-based therapy drugs in the HFS-MAC in the past. Ertapenem is a promising drug for novel combination therapies for MAC lung disease.

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AIMS: The hollow­fibre system for tuberculosis (HFS­TB) is a preclinical model qualified by the European Medicines Agency to underpin the anti­TB drug development process. It can mimic in vivo pharmacokinetic (PK)­pharmacodynamic (PD) attributes of selected antimicrobials, which could feed into in silico models to inform the design of clinical trials. However, historical data and published protocols are insufficient and omit key information to allow experiments to be reproducible. Therefore, in this work, we aim to optimize and standardize various HFS­TB operational procedures. METHODS: First, we characterized bacterial growth dynamics with different types of hollow­fibre cartridges, Mycobacterium tuberculosis strains and media. Second, we mimicked a moxifloxacin PK profile within hollow­fibre cartridges, in order to check drug­fibres compatibility. Lastly, we mimicked the moxifloxacin total plasma PK profile in human after once daily oral dose of 400 mg to assess PK­PD after different sampling methods, strains, cartridge size and bacterial adaptation periods before drug infusion into the system. RESULTS: We found that final bacterial load inside the HFS­TB was contingent on the studied variables. Besides, we demonstrated that drug­fibres compatibility tests are critical preliminary HFS­TB assays, which need to be properly reported. Lastly, we uncovered that the sampling method and bacterial adaptation period before drug infusion significantly impact actual experimental conclusions. CONCLUSION: Our data contribute to the necessary standardization of HFS­TB experiments, draw attention to multiple aspects of this preclinical model that should be considered when reporting novel results and warn about critical parameters in the HFS­TB currently overlooked.

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Rifampicin is recommended for the treatment of Mycobacterium avium complex pulmonary disease alongside azithromycin and ethambutol. We evaluated the azithromycin-ethambutol backbone with and without rifampicin in an intracellular hollow fiber model and performed RNA sequencing to study the differences in adaptation. In an in vitro hollow fiber experiment, we simulated epithelial lining fluid pharmacokinetic profiles of the recommended 3-drug (rifampicin, ethambutol, and azithromycin) or a 2-drug (ethambutol and azithromycin) treatment. THP-1 cells infected with M. avium ATCC700898 were exposed to these regimens for 21 days. We determined intra- and extra-cellular bacterial load- and THP-1 cell densities on days 0, 3, 7, 14, and 21, alongside RNA sequencing. The emergence of macrolide resistance was studied by inoculating intra- and extra-cellular fractions of azithromycin-containing Middlebrook 7H10 agar plates. Complete pharmacokinetic profiles were determined at days 0 and 21. Both therapies maintained stasis of both intra- and extra-cellular bacterial populations for 3 days, whilst regrowth coinciding with the emergence of a macrolide-resistant subpopulation was seen after 7 days. THP-1 cell density remained static. Similar transcriptional profiles were observed for both therapies that were minimally influenced by exposure duration. Transcriptional response was slightly larger during 2-drug treatment. Rifampicin did not add to the antimycobacterial effect to the 2-drug therapy or suppression of emergence resistance. RNA transcription was not greatly altered by the addition of rifampicin, which may be due to strong transcriptional influence of azithromycin and host cells. This questions the role of rifampicin in the currently recommended therapy. These findings should be confirmed in clinical trials.

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Supported liquid membranes have been used to implement a hollow fibre liquid-phase microextraction (HF-LPME) method for the preconcentration of Cd(II) in natural waters as a sample preparation step for its determination by high-resolution continuum source graphite furnace atomic absorption spectrometry (HR-CS-GFAAS). This system was designed to use four hollow fibres simultaneously with the same sample, thus improving the simplicity, speed and reproducibility of the results. The organic liquid membrane bis-(2,4,4-trimethylpentyl) phosphinic acid (Cyanex® 272) dissolved in dihexylether (DHE) was immobilised into the pores of the walls of polypropylene hollow fibres. After extraction, the cadmium-enriched acidic phases were recovered and analysed by triplicate. To optimise the extraction process, the effect of both physical and chemical variables was studied, and optimum results with an enrichment factor (EF) of 292 were obtained for a fibre length of 6 cm, 1.06 M Cyanex 272, 0.04 M HNO3, stirring rate of 600 rpm and an extraction time of 4.26 h. For practical applications, extraction time was reduced to 2 h, keeping the EF as high as 130. Under these conditions, a detection limit of 0.13 ng L-1 Cd(II) was obtained, with a reproducibility of 3.3 % and a linear range up to 3 µg L-1 being achieved. The proposed method was successfully applied to the determination of cadmium in mineral, tap and seawater samples.

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Membrane fouling during the filtration process is a perennial issue and could lead to reduced separation efficiency. In this work, poly(citric acid)-grafted graphene oxide (PGO) was incorporated into a matrix of single-layer hollow fibre (SLHF) and dual-layer hollow fibrr (DLHF) membranes, respectively, aiming to improve membrane antifouling properties during water treatment. Different loadings of PGO ranging from 0 to 1 wt% were first introduced into the SLHF to identify the best PGO loading for the DLHF preparation with its outer layer modified by nanomaterials. The findings showed that at the optimized PGO loading of 0.7 wt%, the resultant SLHF membrane could achieve higher water permeability and bovine serum albumin rejection compared to the neat SLHF membrane. This is due to the improved surface hydrophilicity and increased structural porosity upon incorporation of optimized PGO loading. When 0.7 wt% PGO was introduced only to the outer layer of DLHF, the cross-sectional matrix of the membrane was altered, forming microvoids and spongy-like structures (more porous). Nevertheless, the BSA rejection of the membrane was improved to 97.7% owing to an inner selectivity layer produced from a different dope solution (without the PGO). The DLHF membrane also demonstrated significantly higher antifouling properties than the neat SLHF membrane. Its flux recovery rate is 85%, i.e. 37% better than that of a neat membrane. By incorporating hydrophilic PGO into the membrane, the interaction of the hydrophobic foulants with the membrane surface is greatly reduced.

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Lead (Pb) exposure can be harmful to public health, especially through drinking water. One of the promising treatment methods for lead contaminated water is the adsorption-filtration method. To ensure the cost-effectiveness of the process, naturally derived adsorbent shall be utilised. In this study, hydroxyapatite particles, Ca10(PO4)6(OH)2 (HAP) derived from waste cockle shell, were incorporated into the outer layer of polysulfone/HAP (PSf/HAP) dual-layer hollow fibre (DLHF) membrane to enhance the removal of lead from the water source due to its hydrophilic nature and excellent adsorption capacity. The PSf/HAP DLHF membranes at different HAP loadings in the outer layer (0, 10, 20, 30 and 40 wt%) were fabricated via the co-extrusion phase inversion technique. The performance of the DLHF membranes was evaluated in terms of pure water flux, permeability and adsorption capacity towards lead. The results indicated that the HAP was successfully incorporated into the outer layer of the membrane, as visibly confirmed by microscopic analysis. The trend was towards an increase in pure water flux, permeability and lead adsorption capacity as the HAP loading increased to the optimum loading of 30 wt%. The optimized DLHF membrane displayed a reduced water contact angle by 95%, indicating its improved surface hydrophilicity, which positively affects the pure water flux and permeability of the membrane. Furthermore, the DLHF membrane possessed the highest lead adsorption capacity, 141.2 mg/g. The development of a hybrid inorganic-organic DLHF membrane via the incorporation of the naturally derived HAP in the outer layer is a cost-effective approach to treat lead contaminated water.

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Stem cell and cell therapies, particularly autologous cell therapies, are becoming a common practice. However, in order for these technologies to achieve wide-scale clinical application, the prohibitively high cost associated with these therapies must be addressed through creative engineering. Membranes can be a disruptive technology to reshape the bioprocessing and manufacture of cellular products and significantly reduce the cost of autologous cell therapies. Examples of successful membrane applications include expansions of CAR-T cells, various human stem cells, and production of extracellular vesicles (EVs) using hollow fibre membrane bioreactors. Novel membranes with tailored functions and surface properties and novel membrane modules that can accommodate the changing needs for surface area and transport properties are to be developed to fulfil this key role.

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There is increasing interest in membrane-aerated biofilm reactors (MABRs), due to their energy efficiency and ability to intensify wastewater treatment. While MABR membranes play a key role, supporting biofilms and transferring O2, little research has addressed how membrane types impact MABR performance. This research compared two types of membranes used in commercial MABRs: a silicone hollow-fibre membrane and a 'micromembrane cord,' consisting of an inert cord surrounded by fine proprietary polymeric membranes. We used single-membrane MABRs to determine the oxygen mass transfer coefficient, Km, and explore biofilm development. The silicone membrane had a measured Km of 2.6 m/d, and the micromembrane cord had an apparent Km of 1 m/d. Pure MABR bundles (only biofilm) were operated with synthetic wastewater, and hybrid MABRs (suspended biomass and biofilm) with real wastewater, to explore behaviour for a wide range of conditions. The maximum ammonium oxidation fluxes with synthetic wastewater were 7.8 gN/m2d for the silicone membrane and 4.3 gN/m2d for the micromembrane cord. However, at bulk NH4+ concentrations below 5 mgN/L, the ammonium oxidation fluxes were similar. A previously published MABR model effectively captured the behaviour of each membrane. Nitrification fluxes with real wastewater were lower than synthetic wastewater, likely because of the presence of chemical oxygen demand (COD). Although the ammonium oxidation fluxes were higher for the silicone membranes for a given air supply pressure, the fluxes for the micromembrane cord could be increased using higher intramembrane air pressures. Overall, this research helped understand the impact of membrane types on nitrification fluxes.

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Static concentration in vitro studies have demonstrated that fosfomycin- or sulbactam-based combinations may be efficacious against carbapenem-resistant Acinetobacter baumannii (CRAB). In the present study, we aimed to evaluate the bacterial killing and resistance suppression potential of fosfomycin-sulbactam combination therapies against CRAB isolates in a dynamic infection model. We simulated clinically relevant dosing regimens of fosfomycin (8 g every 8 h, 1 h infusion) and sulbactam (12 g continuous infusion or 4 g every 8 h, 4 h infusion) alone and in combination for 7 days in a hollow-fibre infection model (HFIM) against three clinical isolates of CRAB. The simulated pharmacokinetic profiles in the HFIM were based on fosfomycin and sulbactam data from critically ill patients. Fosfomycin monotherapy resulted in limited bacterial killing. Sulbactam monotherapies resulted in ~ 3 to 4 log10 kill within the first 8 to 32 h followed by regrowth of up to 8 to 10 log10 CFU/mL. A combination of fosfomycin and continuous infusion of sulbactam led to a ~2 to 4 log10 reduction in bacterial burden within the first 24 h, which was sustained throughout the duration of the experiments. A combination of fosfomycin and extended infusion of sulbactam produced a ~4 log10 reduction in colony count within 24 h. This study demonstrated that fosfomycin in combination with sulbactam is a promising option for the treatment of MDR A. baumannii. Further studies are needed to further assess the potential clinical utility of this combination.

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BACKGROUND: Pseudomonas aeruginosa in hospital water is a risk for invasive infection. Point-of-use (POU) filters are used to reduce patient exposure to the organism, and hollow-fibre filters are becoming more popular. However, retrograde colonization of the filter mechanism may contaminate the effluent. AIMS: To assess the efficacy of POU filter head (polysulfone; hollow-fibre matrix) shower filters in preventing the exposure of high-risk patient groups to P. aeruginosa. METHODS: Pre-flush (opening the outlet and collecting the first 100 mL of water) samples were analysed to measure P. aeruginosa contamination from 25 shower outlets (∼21% of all showers on the six wards), with and without a hollow-fibre filter. P. aeruginosa was measured in a subset of outlets harbouring P. aeruginosa (sampling period 19th August 2019 to 10th January 2020). FINDINGS: Water from all 25 showers was heavily colonized [>300 colony-forming units (cfu)/mL] with P. aeruginosa at the showerhead. P. aeruginosa was found in 32% (8/25) of post-filter shower water effluent samples with a geometric mean of 4x106 cfu/mL (N=4) (6.8x104-2x108). Filters were sampled at 15-150 days of use (median 15 days), with 26% (6/23) of filter units becoming colonized before the expiry date. CONCLUSION: POU filter showerhead units may not be effective in preventing exposure of vulnerable patients to P. aeruginosa in hospital water due to retrograde contamination (external contamination of the showerhead passed back to the filter cartridge itself) or failure of the hollow-fibre filter matrix. Reliance should not be placed on the use of hollow-fibre filters to protect patients from exposure to P. aeruginosa without repeated microbiological monitoring.

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Extended-spectrum ß-lactamase (ESBL)-producing Escherichia coli are a global public-health concern. We evaluated the pharmacodynamic activity of piperacillin/tazobactam (TZP) dosing regimens against ESBL-producing versus non-ESBL-producing E. coli. Five E. coli clinical isolates were obtained from Bangladesh. Broth microdilution and whole-genome sequencing (WGS) were performed on the five studied isolates. Three TZP-susceptible ESBL-producing and two non-ESBL-producing E. coli were exposed to TZP regimens of 4.5 g every 6 h (q6h) and every 8 h (q8h) as a 30-min infusion in a dynamic hollow-fibre infection model over 7 days. The extent of bacterial killing was ∼4-5 log10 CFU/mL against ESBL-producing and non-ESBL-producing E. coli with TZP q6h and q8h regimens over the first 8 h. Bacterial killing was similar between two of three ESBL-producing (CTAP#168 and CTAP#169) and two non-ESBL-producing (CTAP#179 and CTAP#180) E. coli clinical isolates over the course of the experiment. ESBL-producing CTAP#173 E. coli was poorly killed (∼1 log) compared with two non-ESBL-producing E. coli over 168 h. WGS revealed that ESBL-producing E. coli isolates co-harboured multiple antimicrobial resistance genes such as blaCTX-M-15, blaEC, blaOXA-1, blaTEM-1 and aac(6')-Ib-cr5. Overall, TZP q6h and q8h dosing regimens attained >3 log bacterial kill against all ESBL-producing or non-ESBL-producing E. coli within 24 h and maintained and prevented the emergence of resistance through the end of the experiment. In conclusion, TZP standard regimens resulted in similar bacterial killing and prevented the emergence of resistance against CTX-M-15-type ESBL-producing and non-ESBL-producing E. coli clinical isolates.

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Low concentrations of pharmaceutically active compounds have been reported in samples from highly complex aqueous environments. Due to their low concentrations, efficient sample pretreatment methods are needed to clean samples and concentrate the compounds of interest prior to instrumental analysis. Hollow fibre liquid-phase microextraction (HF-LPME) is an effective alternative to conventional techniques such as liquid-liquid extraction (LLE) and solid phase extraction (SPE) because it consumes less organic solvent and is less labour intensive with a short extraction time. HF-LPME involves the preconcentration and mass transfer of target analytes from an aqueous sample into an acceptor solution in the lumen of the fibre using a supported liquid membrane (SLM) impregnated in the hollow fibre pores. However, despite the high contaminant selectivity, reproducibility, and enrichment that HF-LPME offers, this technique is limited by membrane instability. Although several advances have been made to address membrane instability, they are either too costly or not feasible for industrial application. Hence, hollow fibre polymer inclusion membrane liquid-phase microextraction (HF-PIM-LPME) was introduced to ameliorate membrane instability. This new approach uses ionic liquids (ILs) as a green solvent, and has demonstrated high membrane stability, good contaminant enrichment, and similar selectivity and reproducibility to HF-SLM-LPME. Hence, this review aims to raise awareness of HF-PIM-LPME as a viable alternative for the selectivity and preconcentration of pharmaceuticals and other contaminants in aquatic environments.

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Electromembranes increase the efficiency of metal transport in liquid-phase microextraction systems by applying an electric potential, which accelerates the transport. Nevertheless, to get high extraction percentages in short extraction times it is necessary to take into account a great variety of factors, and multivariate optimization techniques are the best alternative to determine the most influential variables and to optimize the extraction process. In this work, a fractional factorial design was applied to determine the most influential variables in the extraction of silver by electromembranes. Thus, the effect of tri-isobutylphosphine sulphide (Cyanex 471x) concentration in the organic solution, sodium thiosulphate concentration in the acceptor solution, nitrate concentration in the sample solution, extraction time, stirring rate and electric potential on the enrichment factor were studied. Once the most important variables were selected, a small composite design (Draper-Lin) was used to obtain their optimal values to maximize the enrichment factor. Under these conditions, an experimental enrichment factor of 49.91 ± 3.95 was achieved after 22 min. Finally, the effect of saline matrix on the enrichment factor was tested and the optimized system was successfully applied to analyse silver concentrations at ultratrace levels, within the range of 7-29 ng·L-1 in different real seawater samples.

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BACKGROUND AND OBJECTIVES: Platelet concentrates suspended in a platelet additive solution (PAS-PC) are associated with a reduction in allergic response and are suitable for preparing pathogen-inactivated PC. We aimed to develop an efficient platform for the dual preparation of PAS-PC and platelet-poor plasma. MATERIALS AND METHODS: PAS-PC was prepared in six steps by using a hollow-fibre system based on cross-flow filtration: priming, loading PC, loading PAS, collection of filtered liquid (flow-through) and collection of platelets by washing with PAS followed by washing with air. In this study, the efficacy of platelet and plasma protein recovery and characteristics of recovered PAS-PC and flow-through plasma were analysed in detail. RESULTS: Recoveries of platelet in PAS-PC and plasma protein in the flow-through were 95.4% ± 3.7% and 61.6% ± 5.0%, respectively. The residual plasma protein in PAS-PC was 34.1% ± 2.8%. Although the expression level of CD62P, a platelet activation marker, in recovered platelets was approximately 1.2-fold of that in original platelets, swirling patterns were well retained, and aggregation in PAS-PC was not visible. Agonist-induced aggregabilities, platelet morphology and hypotonic shock recovery were conserved. The patterns of plasma protein and lipoprotein in the flow-through were comparable with those in the original PCs. The multimeric pattern analysis of VWF remained unaltered. CONCLUSION: We propose a highly efficient preparation system that enables the simultaneous production of PAS-PC and platelet-poor plasma. It also achieves a high recovery of functionally well-retained platelets with very low activation.

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Nitrogen oxide (NOx) emissions cause significant impacts on the environment and must therefore be controlled even more stringently. This requires the development of cost-effective removal strategies which simultaneously create value-added by-products or energy from the waste. This study aims to treat gaseous nitric oxide (NO) by hollow-fibre membrane biofilm reactor (HFMBfR) in the presence of nitrite (NO2-) and evaluate nitrous oxide (N2O) emissions formed as an intermediate product during the denitrification process. Accumulated N2O can be utilised in methane oxidation as an oxidant to produce energy. In the first stage of the study, the HFMBfR was operated by feeding only gaseous NO as the nitrogen source. During this period, the best performance was achieved with 92% NO removal efficiency (RE). In the second stage, both NO gas and NO2- were supplied to the system, and 91% NO and 99% NO2- reduction were achieved simultaneously with the maximum N2O generation of 386 ± 31 ppm. Lower influent carbon to nitrogen (C/N) ratios, such as 4.5 and 2.0, and higher NO2--N loading rate of 158 mg N day-1 favoured N2O generation. An improved NO removal rate and N2O accumulation were seen with the increasing amount of PO43- in the medium. The 16S rDNA sequencing analysis revealed that Alicycliphilus denitrificans and Pseudomonas putida were the dominant species. The study shows that an HFMBfR can be successfully used to eliminate both NO2- and gaseous NO and simultaneously generate N2O by adjusting the system parameters such as C/N ratio, NO2- and PO43- loading.

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OBJECTIVES: Mycobacterium avium complex (MAC) bacteria can cause chronic pulmonary disease (PD). Current treatment regimens of azithromycin, ethambutol and rifampicin have culture conversion rates of around 65%. Dynamic, preclinical models to assess the efficacy of treatment regimens are important to guide clinical trial development. The hollow fibre system (HFS) has been applied but reports lack experimental details. METHODS: We simulated the human pharmacokinetics of azithromycin, ethambutol and rifampicin both in plasma and epithelial lining fluid (ELF) in a HFS, exposing THP-1 cells infected with M. avium to the triple-drug regimen for 3 weeks. We accounted for drug-drug interactions and protein-binding and provide all laboratory protocols. We differentiated the effects on the intracellular and extracellular mycobacterial population. RESULTS: The antibiotic concentrations in the HFS accurately reflected the time to peak concentration (Tmax), the peak concentration (Cmax) and half-life of azithromycin, rifampicin and ethambutol in plasma and ELF reported in literature. We find that plasma drug concentrations fail to hold the MAC bacterial load static (ΔLog10 CFU/mLControl:Regimen = 0.66 ± 0.76 and 0.45 ± 0.28 at 3 and 21 days); ELF concentrations do hold the bacterial load static for 3 days and inhibit bacterial growth for the duration of the experiment (ΔLog10 CFU/mLControl:Regimen = 1.1 ± 0.1 and 1.64 ± 0.59 at 3 and 21 days). DISCUSSION: In our model, the current therapy against MAC is ineffective, even when accounting for antibiotic accumulation at the site of infection and intracellularly. New treatment regimens need to be developed and be compared with currently recommended regimens in dynamic models prior to clinical evaluation. With the publication of all protocols we aim to open this technology to new users.

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