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关键词： biomarker monitoring and discovery   cell manufacturing   microfabricated sampling platform   online electrospray ionization mass spectrometry  

摘要： Large-scale manufacturing of therapeutic cells requires bioreactor technologies with online feedback control enabled by monitoring of secreted biomolecular critical quality attributes (CQAs). Electrospray ionization mass spectrometry (ESI-MS) is a highly sensitive label-free method to detect and identify biomolecules, but requires extensive sample preparation before analysis, making online application of ESI-MS challenging. We present a microfabricated, monolithically integrated device capable of continuous sample collection, treatment, and direct infusion for ESI-MS detection of biomolecules in high-salt solutions. The dynamic mass spectrometry probe (DMSP) uses a microfluidic mass exchanger to rapidly condition samples for online MS analysis by removing interfering salts, while concurrently introducing MS signal enhancers to the sample for sensitive biomolecular detection. Exploiting this active conditioning capability increases MS signal intensity and signal-to-noise ratio. As a result, sensitivity for low-concentration biomolecules is significantly improved, and multiple proteins can be detected from chemically complex samples. Thus, the DMSP has significant potential to serve as an enabling portion of a novel analytical tool for discovery and monitoring of CQAs relevant to therapeutic cell manufacturing.

关键词： Dissolved organic nitrogen (DON)   Bioavailable dissolved organic nitrogen (ABDON)   Membrane bioreactor (MBR)   Posttreatment   Eutrophication  

摘要： Bioavailable dissolved organic nitrogen (ABDON) is the component of dissolved organic nitrogen (DON) which supports the growth of algae. Previous research indicates that a membrane bioreactor (MBR) is effective in reducing DON, however, its ability to remove ABDON remains unknown. The present study investigated three full-scale MBRs (membrane type: hydrophilic polyvinylidene fluoride and membrane pore size: 0.04-0.1 mu m) as posttreatment for the removal of ABDON. Results showed that the concentrations of ABDON were not significantly different between influent and effluent at each MBR (p=0.067-0.614, t-test). Analysis of DON molecular composition via ultrahigh resolution mass spectrometry provides supporting evidence that bacterial biomass produced/released ABDON during the biological processes, which would be one of the possible reasons for the low removal efficiency of ABDON (< 0%-14.0%) occurred in the MBR process. Overall, MBRs as posttreatments would have a smaller-than expected impact on primary productivity in receiving waters since a substantial fraction of DON stimulating algal growth cannot be removed by this treatment.

关键词： biomedical equipment  

摘要： Ultrasonic standing wave systems have previously been used for the generation of 3D constructs for a range of cell types. In the present study, we cultured cells from the human hepatoma Huh7 cell line in a Bulk Acoustic Wave field and studied their viability, their functions, and their response to the anti-cancer drug, 5 Fluorouracil (5FU). We found that cells grown in the acoustofluidic bioreactor (AFB) expressed no reduction in viability up to 6 h of exposure compared to those cultured in a conventional 2D system. In addition, constructs created in the AFB and subsequently cultured outside of it had improved functionality including higher albumin and urea production than 2D or pellet cultures. The viability of Huh7 cells grown in the ultrasound field to 5FU anti-cancer drug was comparable to that of cells cultured in the 2D system, showing rapid diffusion into the aggregate core. We have shown that AFB formed 3D cell constructs have improved functionality over the conventional 2D monolayer and could be a promising model for anti-cancer drug testing. Published under license by AIP Publishing.

关键词： Anaerobic fluidized membrane bioreactor   Membrane fouling   Mechanical scouring   Granular activated carbon   Membrane bioreactor  

摘要： Effect of mechanical scouring driven by granular activated carbon (GAC) fluidization on membrane fouling was investigated using a laboratory-scaled, fluidized membrane reactor filtering the effluent from anaerobic fluidized bed bioreactor (AFBR) in domestic wastewater treatment. The GAC particles were fluidized by recirculating a bulk solution only through the membrane reactor to control membrane fouling. The membrane fouling was compared with two different feed solutions, effluent taken from a pilot-scaled, AFBR treating domestic wastewater and its filtrate through 0.1-m membrane pore size. The GAC fluidization driven by bulk recirculation through the membrane reactor was very effective to reduce membrane fouling. Membrane scouring under GAC fluidization decreased reversible fouling resistance effectively. Fouling mitigation was more pronounced with bigger GAC particles than smaller ones as fluidized media. Regardless of the fluidized GAC sizes, however, there was limited effect on controlling irreversible fouling caused by colloidal materials which is smaller than 0.1m. In addition, the deposit of GAC particles that ranged from 180 to 500m in size on membrane surface was very significant and accelerated fouling rate. Biopolymers rejected by the membranes were thought to play a role as binding these small GAC particles on membrane surface strongly.

关键词： Bioreactor   Cytotoxicity   Calixarene   5-fluorouracil   P-31 NMR spectroscopy  

摘要： Introduction: The evaluation of drug's cytotoxicity is a crucial step in the development of new pharmacological compounds. 31P NMR can be a tool for toxicological screening, as it enables the study of drugs' cytotoxicity and their effect on cell energy metabolism in a real-time, in a non-invasive and non-destructive way. This paper details a step-by-step protocol to implement a bioreactor system able to maintain cell viability during NMR acquisitions, at high cell densities and for several hours, enabling toxicological evaluation of pharmacological compounds in living cells. Method: HeLa cells were immobilized in agarose gel threads and continuously perfused with oxygenated medium inside a 5mm NMR tube. Signals corresponding to intracellular high-energy phosphorous compounds were continuously monitored by 31P NMR to assess cell energy levels, intracellular pH and intracellular free Mg2+ concentrations ([Mg2+](f)) under control and in the presence of two different cytotoxic drugs, calix-NH2 or 5-fluorouracil (5-FU). Results: The bioreactor system was effective in maintaining cell energy levels as well as intracellular pH and [Mg2+](f) along time, with a good 31P NMR signal to noise ratio. Calix-NH2 and 5-FU decreased cell energy levels by 35% and 39%, respectively, with a negligible increase in intracellular [Mg2+](f), and without affecting intracellular pH. Discussion: The immobilization and perfusion system here detailed, along with 31P NMR, is useful in toxicological evaluation of new pharmacological compounds, enabling the continuous assessment of drugs' effect on energy levels, intracellular pH and [Mg2+](f) in intact cells, for several hours without compromising cell viability.

关键词： acoustic filter   bioreactor   perfusion   transient transfection   virus-like particles  

摘要： BACKGROUND: Transient gene expression has been widely used for virus-like particles (VLP) production in both small- and large-scale systems. The extended gene expression (EGE) technique is based on repeated medium exchanges and retransfections of cell culture to prolong the production phase. The aim of this study was to demonstrate scalability of the approach by operating EGE continuously in a controlled bioreactor to obtain similar results to those achieved with shake flask EGE for VLP production. A standard batch cultivation was also carried out in shake flask. RESULTS: For EGE in shake flasks and in a bioreactor, cell viability was comparable;however, the bioreactor enabled much higher cell densities and specific growth rates than the shake flasks. Owing to this increased cell growth in the bioreactor, the percentage of GFP-positive cells was considerably lower at the end than in the shake flasks. GagGFP VLP titers were similar in both shake flasks and bioreactor. Nanoparticle tracking analysis revealed that the ratio of VLPs/total particles (VLPs and cell vesicles) was higher in the shake flasks than in the bioreactor, possibly due to higher cell densities achieved in the bioreactor. CONCLUSIONS: In this study, EGE methodology was carried out in a bioreactor system for the first time while maintaining GagGFP production titers. Overall, our findings call for further optimization and implementation of common downstream processing steps to improve yield and VLP quality. (c) 2018 Society of Chemical Industry

关键词： Membrane bioreactor   Moving bed membrane bioreactor   Salinity wastewater   Eukarya   Prokarya   Oligotyping  

摘要： A membrane bioreactor and two hybrid moving bed bioreactor-membrane bioreactors were operated for the treatment of variable salinity wastewater, changing in cycles of 6-h wastewater base salinity and 6-h maximum salinity (4.5 and 8.5mScm(-1) electric conductivity, which relate to 2.4 and 4.8gL(-1) NaCl, respectively), under different hydraulic retention times (6, 9.5, and 12h) and total solids concentrations (2500 and 3500mgL(-1)). The evaluation of the performance of the systems showed that COD removal performance was unaffected by salinity conditions, while BOD5 and TN removals were significantly higher in the low-salinity scenario. The microbial community structure showed differences with respect to salinity conditions for Eukarya, suggesting their higher sensitivity for salinity with respect to Prokarya, which were similar at both salinity scenarios. Nevertheless, the intra-OTU distribution of consistently represented OTUs of Eukarya and Prokarya was affected by the different salinity maximums. Multivariate redundancy analyses showed that several genera such as Amphiplicatus (0.01-5.90%), Parvibaculum (0.27-1.19%), Thiothrix (0.30-1.19%), Rhodanobacter (2.81-5.85%), Blastocatella (0.21-2.01%), and Nitrobacter (0.80-0.99%) were positively correlated with BOD5 and TN removal, and the ecological roles of these were proposed. All these genera were substantially more represented under low-salinity conditions (10-500% higher relative abundance), demonstrating that they might be of importance for the treatment of variable salinity wastewater. Evaluation of Eukarya OTUs showed that many of them lack a consistent taxonomic classification, which highlights the lack of knowledge of the diversity and ecological role of Eukaryotes in saline wastewater treatment processes. The results obtained will be of interest for future design and operation of salinity wastewater treatment systems particularly because little is known on the effect of variable salinity conditions in wastew

关键词： Membrane bioreactor (MBR)   Full-scale application   Wastewater treatment   Cost analysis   Membrane fouling  

摘要： Membrane bioreactor (MBR) technology for wastewater treatment has been developed for over three decades. Our latest survey shows that MBR applications for wastewater treatment are still in rapid growth today. This review summarizes the pros, cons and progress in full-scale MBR applications. Critical statistics on the capital cost, operating cost, footprint, energy consumption and chemical consumption of full-scale MBRs are provided, and are compared to those of conventional activated sludge (CAS) processes with/without tertiary treatment. The efficiencies in full-scale treatment of ordinary pollutants (C, N and P), pathogens (bacteria and viruses) and emerging pollutants (e. g., trace organic pollutants) are reviewed. The long-term operation stability of full-scale MBRs is also discussed with several examples provided, with special attention placed on the seasonal variation of membrane fouling. Finally, the future challenges of MBR application are outlined from the perspectives of fouling control, pollutant removal, cost-effectiveness and competitiveness in specific fields of application.

关键词： Biofilms  

摘要： Methyl tert-butyl ether (MTBE) degradation technologies based on two-phase partitioning systems such as extractive membrane biofilm reactors (EMBFR) permit separation of biological and contaminant compartments, thus allowing optimization of the biological section. In this study, we set-up an EMBFR with three MTBE-degrading and cooperating strains (termed social biofilm: Agrobacterium sp. MS2, Paenibacillus etheri SH7(T) and Rhodococcus ruber EE6). The removal efficiency of the social-biofilm EMBFR was 80%, and functional stability was observed in the reactor, i.e. more efficient than previous studies (single-strain inoculated EMBFR, <50% removal efficiency and unstable function). Metabolite tert-butyl alcohol was not observed, and the EC50 values were higher than those observed in single-strain EMBFRs. Comparative analysis of the MTBE enzymatic pathway and the social-biofilm was performed, where the mechanism of cooperation observed within the social-biofilm is likely due to enzymatic redundancy. Functional outcomes were equal to previous batch tests, hence 100% scalability was obtained. Overall, higher functional and stability outcomes are obtained with the use of the social-biofilm in an MTBE-EMBFR. (C) 2018 Elsevier Ltd. All rights reserved.

关键词： Membrane electro-bioreactor   Membrane fouling   Electrokinetics   Mixed liquor properties   Microbial community   Relationship analysis  

摘要： A membrane electro-bioreactor (MEBR) embracing biological treatment, electrokinetic phenomena and membrane filtration was established by applying intermittent direct current (DC) to MBR. MEBR exhibited significant improvement of treatment performance and reduction of membrane fouling. COD and total phenols removal efficiencies increased to 83.53% and 93.28% at an exposure mode of 24'-OFF/6'-ON, compared to 71.24% and 82.43% in MBR. Trans-membrane pressure increment rate declined dramatically in MEBR, which was mainly attributed to the increase of sludge floc size and decrease of zeta potential, soluble microbial products and specific resistance to filtration, resulted from electrokinetic effects such as electrocoagulation, electrophoresis, electroosmosis and electromigration of ions. It was notable that DC exposure exerted distinct evolution on microbial community, with the improvement of microbial community richness and diversity. The relative abundances of functional genera were promoted noticeably in MEBR. An interactive relevance existed among microbial community structure, mixed liquor properties and operational parameters.