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At least 595 records · Page 33

Data for Protoplast Fusion as a Strategy to Increase Ploidy in Rhodotorula toruloides for Strain Development

Rhodotorula toruloides is a red oleaginous yeast with growing commercial interest because of its hardiness and exceptional lipid production capacity. Because it is a basidiomycete yeast with a complex life cycle, many of the classical breeding methods used with ascomycetes are unavailable for strain improvement. However, we have been able to construct polyploid yeast by fusing protoplasts of parents with the same mating type. Fusing of Y-6985 (A2) and Y-48190 (A2), which had been transformed with complementary antibiotic markers, led to the recovery of two diploids and one triploid. The stability of the fusion yeasts was tested by plating them on non-selective medium after several growth cycles under antibiotics and then testing five colonies per strain for nuclear DNA contents using flow cytometry and standard cell cycle analysis: the triploid and one diploid were stable. Fusants inherited their mitochondria from a single parent, which was demonstrated using restriction fragment length polymorphism (RFLP) of mitochondrial DNA. The phenotypic properties of the parents and fusants were compared in glucose fed-batch bioreactor studies and cellulosic sugar batch cultures. The final lipid titers for the fed-batch cultures were 24.9–39.7 g/L with Y-6985 and the diploid and triploid performing the best and worst, respectively. The fusants demonstrated intermediate hardiness for growth on hydrolysate prepared with dilute-acid pretreated switchgrass and were outperformed by Y-48190. Unlike one of the haploid parents, the fusants grew in 70% v/v concentrated hydrolysate. However, they did not grow as fast as the other haploid. In this study, a modernized protoplast fusion method is resurrected a useful tool for strain development in this yeast, which is complementary with other available methods.

FOS: Biological sciences↗

High-Collection-Efficiency Fluorescence Detection Cell

A new fluorescence cell has been developed for the laser induced fluorescence (LIF) detection of formaldehyde. The cell is used to sample a flow of air that contains trace concentrations of formaldehyde. The cell provides a hermetically sealed volume in which a flow of air containing formaldehyde can be illuminated by a laser. The cell includes the optics for transmitting the laser beam that is used to excite the formaldehyde and for collecting the resulting fluorescence. The novelty of the cell is its small size and simple design that provides a more robust and cheaper alternative to the state of the art. Despite its simplicity, the cell provides the same sensitivity to detection as larger, more complicated cells.

Hanisco, Thomas↗

Ionospheric convection signatures and magnetic field topology

A statistical study of signatures of the high-latitude ionospheric convection pattern and the simultaneously observed energetic electron precipitation is presented. Most often found are convection cells in which the sunward flowing region contains auroral particle precipitation but the antisunward flowing region does not. However, observations also show the frequent occurrence of convection cells in which neither the antisunward nor the sunward flowing plasma region contains auroral particle precipitation. These findings may appear within the dawnside or duskside convection pattern and strongly suggest that such convection cells may be associated with open magnetic field lines that thread the magnetotail lobes.

Coley, W. R.↗

Computer modeling of high-efficiency solar cells

Transport equations which describe the flow of holes and electrons in the heavily doped regions of a solar cell are presented in a form that is suitable for device modeling. Two experimentally determinable parameters, the effective bandgap shrinkage and the effective asymmetry factor are required to completely model the cell in these regions. Nevertheless, a knowledge of only the effective bandgap shrinkage is sufficient to model the terminal characteristics of the cell. The results of computer simulations of the effects of heavy doping are presented. The insensitivity of the terminal characteristics to the choice of effective asymmetry factor is shown along with the sensitivity of the electric field and quasielectric fields to this parameter. The dependence of the terminal characteristics on the effective bandgap shrinkage is also presented.

Schartz, R. J.↗

Proton Exchange Membrane Fuel Cell Transient Load Response

Spaceflight mission profiles may not be limited to constant power loads. It is necessary for the associated power system to support changes in load in addition to being energy dense. H2-O2 proton exchange membrane (PEM) fuel cells can accommodate such requirements without significant current smoothing or additional devices to provide transient power. This testing involved current load step changes to characterize non-flow-through PEM fuel cell response to increasing and decreasing load step changes over a range of 0 to 0.5 A/sq. cm. It was determined that fuel cell output voltage stabilizes within 4 ms with no noticeable steady-state performance degradation, following exposure of the fuel cell to transient loads of up to ~1.5 A/sq. cm. Faster stabilization generally occurred following larger step changes.

fuel cell↗

Rapid native single cell mass spectrometry

A method for analyzing single cells by mass spectrometry includes the steps of providing a plurality of cells in a liquid medium and placing the cells and liquid medium in a single cell isolation and ejection system. Liquid medium containing a single cell is released from the single cell isolation and ejection system. The liquid medium and single cell are captured in a capture probe containing a flowing capture probe solvent. The cell is lysed by a lysis inducer in the capture probe to disperse single cell components into the medium. The lysed single cell components are transported to a mass spectrometer, where the lysed single cell components entering the mass spectrometer are spatially and temporally separated from any dispersed components of another single cell from the sample entering the mass spectrometer. Mass spectrometry is conducted on the lysed single-cell components. A system for analyzing single cells by mass spectrometry is also disclosed.

Cahill, John F.↗

Rapid native single cell mass spectrometry

A method for analyzing single cells by mass spectrometry includes the steps of providing a plurality of cells in a liquid medium and placing the cells and liquid medium in a single cell isolation and ejection system. Liquid medium containing a single cell is released from the single cell isolation and ejection system. The liquid medium and single cell are captured in a capture probe containing a flowing capture probe solvent. The cell is lysed by a lysis inducer in the capture probe to disperse single cell components into the medium. The lysed single cell components are transported to a mass spectrometer, where the lysed single cell components entering the mass spectrometer are spatially and temporally separated from any dispersed components of another single cell from the sample entering the mass spectrometer. Mass spectrometry is conducted on the lysed single-cell components. A system for analyzing single cells by mass spectrometry is also disclosed.

Cahill, John F.↗

Tuning Alkaline Anion Exchange Membranes through Crosslinking: A Review of Synthetic Strategies and Property Relationships

Alkaline anion exchange membranes (AAEMs) are an enabling component for next-generation electrochemical devices, including alkaline fuel cells, water and CO2 electrolyzers, and flow batteries. While commercial systems, notably fuel cells, have traditionally relied on proton-exchange membranes, hydroxide-ion conducting AAEMs hold promise as a method to reduce cost-per-device by enabling the use of non-platinum group electrodes and cell components. AAEMs have undergone significant material development over the past two decades; however, challenges remain in the areas of durability, water management, high temperature performance, and selectivity. In this review, we survey crosslinking as a tool capable of tuning AAEM properties. While crosslinking implementations vary, they generally result in reduced water uptake and increased transport selectivity and alkaline stability. We survey synthetic methodologies for incorporating crosslinks during AAEM fabrication and highlight necessary precautions for each approach.

08 HYDROGEN↗

Improved fuel cell

Concept in fuel cell design has two cells, each with its own reactant flow fields bonded into single modular unit. Cells require fewer parts and occupy less space than conventional fuel cells, effecting easier assembly and maintenance.

Bell, W. F.↗

CARS Temperature Measurements in a Hypersonic Propulsion Test Facility

Nonintrusive diagnostic measurements were performed in the supersonic reacting flow of the Hypersonic Propulsion Test Cell 2 at NASA-Langley. A Coherent Anti-stokes Raman Spectroscopy (CARS) system was assembled specifically for the test cell environment. System design considerations were: (1) test cell noise and vibration; (2) contamination from flow field or atmospheric borne dust; (3) unwanted laser or electrically induced combustion (inside or outside the duct); (4) efficient signal collection; (5) signal splitting to span the wide dynamic range present throughout the flow field; (6) movement of the sampling volume in the flow; and (7) modification of the scramjet model duct to permit optical access to the reacting flow with the CARS system. The flow in the duct was a nominal Mach 2 flow with static pressure near one atmosphere. A single perpendicular injector introduced hydrogen into the flow behind a rearward facing step. CARS data was obtained in three planes downstream of the injection region. At least 20 CARS data points were collected at each of the regularly spaced sampling locations in each data plane. Contour plots of scramjet combustor static temperature in a reacting flow region are presented.

Jarrett, Olin, Jr.↗

Cell-Mediated Immune Function and Cytokine Regulation During Space Flight

The changes in immune function which occur during space flight potentially expose the crews to an increased risk for development of illness. Decreased cellular immune function has been repeatedly documented after space flight and confirmed during flight by in vivo delayed-type hypersensitivity testing. However, correlation of immune changes with a clinically significant risk factor has not yet been performed. Our hypothesis is that space flight induces a decrease in cell-mediated immune function accompanied by a shift from a type 1 cytokine pattern (favoring cell-mediated immunity) to a type 2 cytokine pattern (favoring humoral immunity). We further hypothesize that reactivation of latent viruses will occur during space flight in association with the decreased cellular immunity. To test these hypotheses, we will determine the effects of space flight on cell-mediated immunity and viral reactivation. We will utilize delayed-type hypersensitivity testing as an in vivo measure of integrated cell-mediated immune function. The production of cytokines and immunoregulatory factors by lymphocytes and monocytes will be measured to determine whether changes in cytokine patterns are associated with the space flight-induced immune dysregulation. Correlation of antigen-specific immune changes with reactivation of latent herpes viruses will be determined by measuring peripheral levels of viral (CMV, VZV, EBV) antigen-specific T cells and comparing to the levels of EBV-infected B-cells by fluorescence in situ hybridization and flow cytometry. A comparison of cell-mediated immune function, cytokine regulation and viral reactivation will provide new insights into crew member health risks during flight.

Sams, Clarence F.↗

Ground Test Stand Development for One-step Electrochemical Production of High-Pressure Oxygen

NASA designed a test rig, High-Pressure Oxygen Generation Assembly (HPOGA) to test a novel method of electrolysis as an alternative technology to the state-of-the-art Oxygen Generation Assembly (OGA) currently on the International Space Station (ISS). This approach involves utilizing a specialized electrolysis cell stack designed to both generate and compress high-pressure oxygen up to 3600 PSIG. The design eliminates the need for an external compressor and hydrogen separator, streamlining the oxygen generation process. HPOGA tests the viability of an electrolysis cell stack for extended life usage. Within the system, there are several key subsystems: nitrogen, high/low pressure oxygen, hydrogen, water recirculation, and water injection. The water injection subsystem supplies deionized (DI) water to a centralized reservoir for consumption. Subsequently, a water recirculation loop transfers the DI water across the cathode side of the electrolysis cell stack with constant flow rate. The result is a supply of hydrogen in the water loop and oxygen accumulation on the anode side of the cell stack. Hydrogen is evaporated out as part of the hydrogen sub-system which also measures hydrogen/oxygen concentration for safety. The oxygen subsystem is responsible for managing high pressure oxygen, O2 exhaust, and conducting hydrogen/oxygen concentration measurements. The nitrogen subsystem serves dual functions, purging the hydrogen/oxygen subsystems, and regulating pressure in both high/low-pressure O2 sections via pneumatic back pressure regulators. Various sensors monitor pressure, temperature, humidity, gas concentration and flow rate in each subsystem. This paper presents the design and testing of a comprehensive test stand, which serves to operate and functionally verify the performance of the electrolysis cell while offering valuable lessons learned.

electrolysis↗

Experimental investigation of recirculating cells in laminar coaxial jets.

Utilizing several unique means of introducing smoke into the flow field for careful visualization in addition to hot-wire techniques, experiments are performed in a specially designed facility producing laminar flows up to considerably high Reynolds numbers. Characteristics of the cells and the flow conditions that bring them about are documented by smoke photographs in the Reynolds number velocity ratio plane and the results are compared to previous analytical predictions. The cells are found to fall into three categories with different flow characteristics involving unsteadiness in position, and shear layer instabilities which result in higher mixing with the outer streams.-

Warpinski, N. R.↗

Shear and mixing effects on cells in agitated microcarrier tissue culture reactors

Tissue cells are known to be sensitive to mechanical stresses imposed on them by agitation in bioreactors. The amount of agitation provided in a microcarrier or suspension bioreactor should be only enough to provide effective homogeneity. Three distinct flow regions can be identified in the reactor: bulk turbulent flow, bulk laminar flow and boundary-layer flows. Possible mechanisms of cell damage are examined by analyzing the motion of microcarriers or free cells relative to the surrounding fluid, to each other and to moving or stationary solid surfaces. The primary mechanisms of cell damage appear to result from: (1) direct interaction between microcarriers and turbulent eddies; (2) collisions between microcarriers in turbulent flow; and (3) collisions against the impeller or other stationary surfaces. If the smallest eddies of turbulent flow are of the same size as the microcarrier beads, they may cause high shear stresses on the cells. Eddies the size of the average interbead spacing may cause bead-bead collisions which damage cells. The severity of the collisions increases when the eddies are also of the same size as the beads. Impeller collisions occur when beads cannot avoid the impeller leading edge as it advances through the liquid. The implications of the results of this analysis on the design and operation of tissue culture reactors are discussed.

Cherry, Robert S.↗

Electrophoresis experiment. Experiment MA-014

A continuous free-flow electrophoresis study was conducted during the Apollo-Soyuz Test Project Mission to investigate and evaluate the increase in sample flow rate and sample resolution achievable in space. The electrophoresis equipment was designed for the separation of four mixtures of biological cells with variable sample flow rates, buffer flow rates, and electric field gradients. Separation quality was assessed by measuring the light from a quartz lamp through the electrophoresis channel and onto a photodiode system. The data evaluation indicates that all monitored systems operated correctly during the experiment. The optical system produced a light that was too bright to discern true cell distributions, but final analysis of scientific data by computer processing shows the expected distribution of separated cells.

Hannig, K. H.↗

Electrophoresis experiment, experiment MA-014

A continuous, free flow electrophoresis study was conducted during the Apollo Soyuz Test Project mission to investigate and evaluate the increase in sample flow rate and sample resolution achievable in space. The electrophoresis equipment was designed for the separation of four mixtures of biological cells with variable sample flow rates, buffer flow rates, and electric field gradients. Separation quality was assessed by measuring the light from a quartz lamp through the electrophoresis channel and on to a photodiode system. The preliminary results indicate that all monitored systems operated correctly during the experiment. The optical system produced a light that was too bright to discern true cell distributions, but data were received that show a distribution of separated cells.

Hannig, K.↗