scholarly journals Production of PHB From CO2-Derived Acetate With Minimal Processing Assessed for Space Biomanufacturing

2021 ◽  
Vol 12 ◽  
Author(s):  
Stefano Cestellos-Blanco ◽  
Skyler Friedline ◽  
Kyle B. Sander ◽  
Anthony J. Abel ◽  
Ji Min Kim ◽  
...  
Keyword(s):  
Durable Goods ◽  
Life Support ◽  
Culture Media ◽  
Minimal Processing ◽  
Cell Culture Media ◽  
Support Materials ◽  
Mission Success ◽  
Phb Production ◽  
Sporomusa Ovata

Providing life-support materials to crewed space exploration missions is pivotal for mission success. However, as missions become more distant and extensive, obtaining these materials from in situ resource utilization is paramount. The combination of microorganisms with electrochemical technologies offers a platform for the production of critical chemicals and materials from CO2 and H2O, two compounds accessible on a target destination like Mars. One such potential commodity is poly(3-hydroxybutyrate) (PHB), a common biopolyester targeted for additive manufacturing of durable goods. Here, we present an integrated two-module process for the production of PHB from CO2. An autotrophic Sporomusa ovata (S. ovata) process converts CO2 to acetate which is then directly used as the primary carbon source for aerobic PHB production by Cupriavidus basilensis (C. basilensis). The S. ovata uses H2 as a reducing equivalent to be generated through electrocatalytic solar-driven H2O reduction. Conserving and recycling media components is critical, therefore we have designed and optimized our process to require no purification or filtering of the cell culture media between microbial production steps which could result in up to 98% weight savings. By inspecting cell population dynamics during culturing we determined that C. basilensis suitably proliferates in the presence of inactive S. ovata. During the bioprocess 10.4 mmol acetate L –1 day–1 were generated from CO2 by S. ovata in the optimized media. Subsequently, 12.54 mg PHB L–1 hour–1 were produced by C. basilensis in the unprocessed media with an overall carbon yield of 11.06% from acetate. In order to illustrate a pathway to increase overall productivity and enable scaling of our bench-top process, we developed a model indicating key process parameters to optimize.

scholarly journals In Situ Swelling Behavior of Chitosan-Polygalacturonic Acid/Hydroxyapatite Nanocomposites in Cell Culture Media

2010 ◽  
Vol 2010 ◽  
pp. 1-12 ◽  
Author(s):  
Rohit Khanna ◽  
Kalpana S. Katti ◽  
Dinesh R. Katti
Keyword(s):  
Cell Culture ◽  
Structural Integrity ◽  
Culture Media ◽  
Sample Surface ◽  
Nanocomposite Films ◽  
Apatite Formation ◽  
Polygalacturonic Acid ◽  
Cell Culture Media ◽  
Polyelectrolyte Complexes

The molecular and mechanical characteristics of in situ degradation behavior of chitosan-polygalacturonic acid/hydroxyapatite (Chi-PgA-HAP) nanocomposite films is investigated using Fourier Transform Infrared spectroscopy (FTIR), Atomic Force Microscopy (AFM), and modulus mapping techniques for up to 48 days of soaking in cell culture media. The surface molecular structure of media-soaked samples changes over the course of 48 days of soaking, as indicated by significant changes in phosphate vibrations (1200–900 ) indicating apatite formation. Chitosan-Polygalacturonic acid polyelectrolyte complexes (PECs) govern structural integrity of Chi-PgA-HAP nanocomposites and FTIR spectra indicate that PECs remain intact until 48 days of soaking. In situ AFM experiments on media-soaked samples indicate that soaking results in a change in topography and swelling proceeds differently at the initial soaking periods of about 8 days than for longer soaking. In situ modulus mapping experiments are done on soaked samples by probing 1–3 nm of surface indicating elastic moduli of 4 GPa resulting from proteins adsorbed on Chi-PgA-HAP nanocomposites. The elastic modulus decreases by 2 GPa over a long exposure to cell culture media (48 days). Thus, as water enters the Chi-PgA-HAP sample, surface molecular interactions in Chi-PgA-HAP structure occur that result in swelling, causing small changes in nanoscale mechanical properties.

Investigation of Near-Infrared Spectroscopy for Periodic Determination of Glucose in Cell Culture Media in Situ

2000 ◽  
Vol 54 (10) ◽  
pp. 1453-1457 ◽  
Author(s):  
Christopher B. Lewis ◽  
Roger J. McNichols ◽  
Ashok Gowda ◽  
Gerard L. Coté
Keyword(s):  
Cell Culture ◽  
Infrared Spectroscopy ◽  
Near Infrared Spectroscopy ◽  
Near Infrared ◽  
Culture Media ◽  
Cell Culture Media

High-pressure freezing of cells in suspension for low-voltage scanning electron microscopy (LVSEM)

1991 ◽  
Vol 49 ◽  
pp. 64-65
Author(s):  
Marek Malecki ◽  
James Pawley ◽  
Hans Ris
Keyword(s):  
Electron Microscopy ◽  
High Pressure ◽  
Low Voltage ◽  
Culture Media ◽  
Cell Structure ◽  
Freeze Substitution ◽  
Ice Crystal ◽  
High Pressure Freezing ◽  
Cell Culture Media ◽  
Voltage Scanning

The ultrastructure of cells suspended in physiological fluids or cell culture media can only be studied if the living processes are stopped while the cells remain in suspension. Attachment of living cells to carrier surfaces to facilitate further processing for electron microscopy produces a rapid reorganization of cell structure eradicating most traces of the structures present when the cells were in suspension. The structure of cells in suspension can be immobilized by either chemical fixation or, much faster, by rapid freezing (cryo-immobilization). The fixation speed is particularly important in studies of cell surface reorganization over time. High pressure freezing provides conditions where specimens up to 500μm thick can be frozen in milliseconds without ice crystal damage. This volume is sufficient for cells to remain in suspension until frozen. However, special procedures are needed to assure that the unattached cells are not lost during subsequent processing for LVSEM or HVEM using freeze-substitution or freeze drying. We recently developed such a procedure.

Cell culture media: NMR approaches to evaluating quality and nutrient consumption

Planta Medica ◽  
2016 ◽  
Vol 81 (S 01) ◽  
pp. S1-S381
Author(s):  
KB Killday ◽  
AS Freund ◽  
C Fischer ◽  
KL Colson
Keyword(s):  
Cell Culture ◽  
Culture Media ◽  
Cell Culture Media ◽  
Nutrient Consumption

The Influence of Glycosylation on the Catalytic and Fibrinolytic Properties of Pro-Urokinase

1992 ◽  
Vol 68 (05) ◽  
pp. 539-544 ◽  
Author(s):  
Catherine Lenich ◽  
Ralph Pannell ◽  
Jack Henkin ◽  
Victor Gurewich
Keyword(s):  
Escherichia Coli ◽  
Mammalian Cell ◽  
Human Gene ◽  
Culture Media ◽  
Mammalian Cell Culture ◽  
Glycosylation Site ◽  
Clot Lysis ◽  
Cell Culture Media ◽  
E Coli ◽  
The Uk

SummaryWe previously found that human pro-UK expressed in Escherichia coli is more active in fibrinolysis than recombinant human pro-UK obtained from mammalian cell culture media. To determine whether this difference is related to the lack of glycosylation of the E. coli product, we compared the activity of E. coli-derived pro-UK [(-)pro-UK] with that of a glycosylated pro-UK [(+)pro-UK] and of a mutant of pro-UK missing the glycosylation site at Asn-302 [(-) (302) pro-UK]. The latter two pro-UKs were obtained by expression of the human gene in a mammalian cell. The nonglycosylated pro-UKs were activated by plasmin more efficiently (≈2-fold) and were more active in clot lysis (1.5-fold) than the (+)pro-UK. Similarly, the nonglycosylated two-chain derivatives (UKs) were more active against plasminogen and were more rapidly inactivated by plasma inhibitors than the (+)UK.These findings indicate that glycosylation at Asn-302 influences the activity of pro-UK/UK and could be the major factor responsible for the enhanced activity of E. coli-derived pro-UK.

In situ deteriorating patient simulation in general practice

2020 ◽  
Vol 70 (suppl 1) ◽  
pp. bjgp20X711425
Author(s):  
Joanna Lawrence ◽  
Petronelle Eastwick-Field ◽  
Anne Maloney ◽  
Helen Higham
Keyword(s):  
Life Support ◽  
Early Recognition ◽  
Phase 1 ◽  
Patient Simulation ◽  
Medical Emergency ◽  
Phase 2 ◽  
Action Plans ◽  
Integrated Simulation ◽  
Deteriorating Patient

BackgroundGP practices have limited access to medical emergency training and basic life support is often taught out of context as a skills-based event.AimTo develop and evaluate a whole team integrated simulation-based education, to enhance learning, change behaviours and provide safer care.MethodPhase 1: 10 practices piloted a 3-hour programme delivering 40 minutes BLS and AED skills and 2-hour deteriorating patient simulation. Three scenarios where developed: adult chest pain, child anaphylaxis and baby bronchiolitis. An adult simulation patient and relative were used and a child and baby manikin. Two facilitators trained in coaching and debriefing used the 3D debriefing model. Phase 2: 12 new practices undertook identical training derived from Phase 1, with pre- and post-course questionnaires. Teams were scored on: team working, communication, early recognition and systematic approach. The team developed action plans derived from their learning to inform future response. Ten of the 12 practices from Phase 2 received an emergency drill within 6 months of the original session. Three to four members of the whole team integrated training, attended the drill, but were unaware of the nature of the scenario before. Scoring was repeated and action plans were revisited to determine behaviour changes.ResultsEvery emergency drill demonstrated improved scoring in skills and behaviour.ConclusionA combination of: in situ GP simulation, appropriately qualified facilitators in simulation and debriefing, and action plans developed by the whole team suggests safer care for patients experiencing a medical emergency.

scholarly journals Simulation and Analysis of Self-Replicating Robot Decision-Making Systems

Computers ◽  
2021 ◽  
Vol 10 (1) ◽  
pp. 9
Author(s):  
Andrew Jones ◽  
Jeremy Straub
Keyword(s):  
Lower Risk ◽  
System Construction ◽  
System Configuration ◽  
Operating Environment ◽  
Robot System ◽  
Robotic Exploration ◽  
Mission Success ◽  
Robot Systems ◽  
System Configurations

Self-replicating robot systems (SRRSs) are a new prospective paradigm for robotic exploration. They can potentially facilitate lower mission costs and enhance mission capabilities by allowing some materials, which are needed for robotic system construction, to be collected in situ and used for robot fabrication. The use of a self-replicating robot system can potentially lower risk aversion, due to the ability to potentially replenish lost or damaged robots, and may increase the likelihood of mission success. This paper proposes and compares system configurations of an SRRS. A simulation system was designed and is used to model how an SRRS performs based on its system configuration, attributes, and operating environment. Experiments were conducted using this simulation and the results are presented.

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