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A novel, alternative intensified cell culture process comprised of a linked bioreactor system is presented. An N-1 perfusion bioreactor maintained cells in a highly proliferative state and provided a continuous inoculum source to a second bioreactor operating as a continuous-flow stirred-tank reactor (CSTR). An initial study evaluated multiple system steady-states by varying N-1 steady-state viable cell densities, N-1 to CSTR working volume ratios, and CSTR dilution rates. After identifying near optimum system steady-state parameters yielding a relatively high volumetric productivity while efficiently consuming media, a subsequent lab-scale experiment demonstrated the startup and long-term operation of the envisioned manufacturing process for 83 days. Additionally, to compensate for the cell-specific productivity loss over time due to cell line instability, the N-1 culture was also replaced with younger generation cells, without disturbing the steady-state of the system. Using the model cell line, the system demonstrated a two-fold volumetric productivity increase over the commercial-ready, optimized fed-batch process.  相似文献   
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Upper airway compliance indicates the potentialof the airway to collapse and is relevant to the pathogenesis ofobstructive sleep apnea. We hypothesized that compliance would varyover the rostral-to-caudal extent of the pharyngeal airway. In aparalyzed isolated upper airway preparation in cats, we controlledstatic upper airway pressure during magnetic resonance imaging (MRI, 0.391-mm resolution). We measured cross-sectional area andanteroposterior and lateral dimensions from three-dimensionalreconstructed MRIs in axial slices orthogonal to the airway centerline.High-retropalatal (HRP), midretropalatal (MRP), and hypopharyngeal(HYP) regions were defined. Regional compliance was significantlyincreased from rostral to caudal regions as follows: HRP < MRP < HYP (P < 0.0001), and compliancedifferences among regions were directly related to collapsibility. Thusour findings in the isolated upper airway of the cat support thehypothesis that regional differences in pharyngeal compliance exist andsuggest that baseline regional variations in compliance andcollapsibility may be an important factor in the pathogenesis andtreatment of obstructive sleep apnea.

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As a pulmonary component of Predictive Studies V, designed to determine O2 tolerance of multiple organs and systems in humans at 3.0-1.5 ATA, pulmonary function was evaluated at 1.0 ATA in 13 healthy men before and after O2 exposure at 3.0 ATA for 3.5 h. Measurements included flow-volume loops, spirometry, and airway resistance (Raw) (n = 12); CO diffusing capacity (n = 11); closing volumes (n = 6); and air vs. HeO2 forced vital capacity maneuvers (n = 5). Chest discomfort, cough, and dyspnea were experienced during exposure in mild degree by most subjects. Mean forced expiratory volume in 1 s (FEV1) and forced expiratory flow at 25-75% of vital capacity (FEF25-75) were significantly reduced postexposure by 5.9 and 11.8%, respectively, whereas forced vital capacity was not significantly changed. The average difference in maximum midexpiratory flow rates at 50% vital capacity on air and HeO2 was significantly reduced postexposure by 18%. Raw and CO diffusing capacity were not changed postexposure. The relatively large change in FEF25-75 compared with FEV1, the reduction in density dependence of flow, and the normal Raw postexposure are all consistent with flow limitation in peripheral airways as a major cause of the observed reduction in expiratory flow. Postexposure pulmonary function changes in one subject who convulsed at 3.0 h of exposure are compared with corresponding average changes in 12 subjects who did not convulse.  相似文献   
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