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We present the development of microbioreactors with a sensitive and accurate optical coupling to a confocal Raman microspectrometer. We show that such devices enable in situ and in vitro investigation of cell cultures for tissue engineering by chemically sensitive Raman spectroscopic imaging techniques. The optical resolution of the Raman microspectrometer allows recognition and chemical analysis of subcellular features. Human bone marrow stromal cells (hBMSCs) have been followed after seeding through a phase of early proliferation until typically 21 days later, well 'after the cells have differentiated to osteoblasts. Long-term perfusion of cells in the dynamic culture conditions was shown to be compatible with experimental optical demands and off-line optical analysis. We show that Raman optical analysis of cells and cellular differentiation in microbioreactors is feasible down to the level of subcellular organelles during development. We conclude that microbioreactors combined with Raman microspectroscopy are a valuable tool to study hBMSC proliferation, differentiation, and development into tissues under in situ and in vitro conditions. 10.1021/ac902515c

A pilot-scale experiment was carried out to study the simultaneous removal of nitrogen and phosphorous from municipal wastewater by an innovative continuous-flow integrated biological reactor (CIBR) process. A three-phase separator was used in the CIBR process, which not only saved energy consumption of sludge returning, but also solved the sludge-gas separating problem. The optimal working condition was 2 h aeration, 1 h agitation, and 1 h settling, with an energy consumption of 0.23 kW h/[m.sup.3]. The average removal of chemical oxygen demand (COD), ammonia nitrogen (N[H.sup.+.sub.4]-N), total nitrogen (TN), and total phosphorus (TP) under the optimal conditions were 72.87, 75.23, 61.25, and 68.25%, respectively. The distributing rules of dissolved oxygen, pH, mixed liquid suspended solid, COD, N[H.sup.+.sub.4]-N, N[O.sup.-.sub.3]-N, TN, and TP in each phase of CIBR was studied. It was indicated that the appropriate condition was created for the simultaneous removal of nitrogen and phosphorus in the integrated reactor. The study demonstrated the feasibility of using CIBR process for simultaneous removal of nitrogen and phosphorus at the average temperature 12.2[degrees]C. DOI: 10.1061/(ASCE)0733-9372(2008)134:3(169) CE Database subject headings: Assets; Sustainable development; Wastewater management; Water quality.

An Escherichia coli cell-free expression system is encapsulated in a phospholipid vesicle to build a cell-like bioreactor. Large unilamellar vesicles containing extracts are produced in an oil-extract emulsion. To form a bilayer the vesicles are transferred into a feeding solution that contains ribonucleotides and amino acids. Transcription-translation of plasmid genes is isolated in the vesicles. Whereas in bulk solution expression of enhanced GFP stops after 2 h, inside the vesicle permeability of the membrane to the feeding solution prolongs the expression for up to 5 h. To solve the energy and material limitations and increase the capacity of the reactor, the [alpha]-hemolysin pore protein from Staphylococcus aureus is expressed inside the vesicle to create a selective permeability for nutrients. The reactor can then sustain expression for up to 4 days with a protein production of 30 [micro]M after 4 days. Oxygen diffusion and osmotic pressure are critical parameters to maintain expression and avoid vesicle burst. [alpha]-hemolysin | cell-free protein expression | membrane-anchoring polypeptide