Results for “gene transfer” — Page 9

NASA NTRS2010

Trade Study for 9 kW Water Membrane Evaporator

Sublimators have been proposed and used in spacecraft for heat rejection. Sublimators are desirable heat rejection devices for short duration use because they can transfer large amounts of heat using little mass and are self-regulating devices. Sublimators reject heat into space by freezing water inside a porous substrate, allowing it to sublimate into vapor, and finally venting it into space. The state of the art thermal control system in orbiting spacecraft is a two loop, two fluid system. The external coolant loop typically uses a toxic single phase fluid that acquires heat from the spacecraft and rejects most of it via a radiator. The sublimator functions as a transient topper for orbiting spacecraft during day pass periods when radiator efficiency decreases. The sublimator interfaces with the internal loop through a built in heat exchanger. The internal loop fluid is non-toxic and is typically a propylene glycol and water solution with inhibitors to prevent corrosion with aluminum fins of the heat exchangers. Feedwater is supplied from a separate line to the sublimator to maintain temperature control of the cabin and vehicle hardware. Water membrane evaporators have been developed for spacecraft and spacesuits. They function similar to a sublimator but require a backpressure valve which could be actuated for this application with a simple fully open or fully closed modes. This technology would be applied to orbital thermal control (lunar or planetary). This paper details a trade study showing that evaporators would greatly reduce the consumable that is used, effectively wasted, by sublimators during start up and shut down during the topping phases of each orbit. State of the art for 9 kW sublimators reject about 870 W per kilogram of mass and 1150 W per liter of volume. If water with corrosion inhibitors is used the evaporators would be about 80% of the mass and volume of the equivalent system. The size and mass increases to about 110% if the internal fluid is 50% propylene glycol/50% water. The true benefit comes from the backpressure valve, that prevents the cyclical shutdown/startup loss of the sublimator and amounts to as much as 0.85 kg per orbit.

Bue, Grant C.↗

NASA NTRS2001

A spontaneously arising mutation in connexin32 with repeated passage of FRTL-5 cells coincides with increased growth rate and reduced thyroxine release

In this study we examine changes in the cellular properties of FRTL-5 cells as a function of passage number, with particular emphasis on gap junction expression, karyotype, morphology, growth rate and thyroxine (T(4)) release. Early passage FRTL-5 follicular cells transfer dye through gap junctions from injected cell(s) to third-order neighboring cells and beyond within their respective follicles and have immuno-detectable connexin32 (Cx32) type gap junctional plaques in their lateral contacting plasma membranes. By contrast, FRTL-5 cells established as monolayers, or as follicles from cultures passed more than 15 times, did not transfer microinjected Lucifer Yellow dye to contiguous neighboring cells and did not express any immuno-detectable rat thyroid specific connexins (Cx43, Cx32 or Cx26). Western blots confirmed that total, membrane and cytosolic Cx32 protein was present only in early pass follicular cultures. To better understand the passage-dependent loss of Cx32 expression, RT-PCR primers were made to the most unique sequences of the rat Cx32 molecule, the cytoplasmic and carboxyl-terminal regions. These primers were used to screen FRTL-5 RNA from cultures of various passage numbers. The results revealed that later passage cultures had a single base deletion in the middle of the Cx32 cytoplasmic loop region at nucleotide position 378. This base deletion was in the middle position of the codon for amino acid 116, which is normally a CAC (histidine) but read with the frame shift was a CCC (proline). The four amino acids that followed this deletion were also altered with the fourth one becoming UAA, the ochre translation stop codon. This premature stopping of translation resulted in a truncation of 60% of the protein, which included the remaining cytoplasmic loop, third and fourth transmembrane regions and the carboxyl-terminus. The later passage cultures did not produce a carboxyl-terminal RT-PCR product, indicating that the mRNA was also truncated. These regions of the Cx32 molecule contain the sequences and epitopes to which probes and antibodies are directed, and as such alterations of these regions with repeated passage explains reports by others that FRTL-5 cells do not express Cx32, and implies that cultures used for these assessments were passed more than 15 times. To determine if genetic or epigenetic abnormalities existed in FRTL-5 cells we performed chromosome spreads from various passage cultures. FRTL-5 cells have been reported to be diploid and more recently non-diploid; however, we found them to be fully tetraploid. This tetraploidy appears to be unstable in that later passes are tetraploid plus two or three extra chromosomes. There were no obvious translocations, breaks or large-scale interstitial deletions of any chromosomes in the FRTL-5 cultures tested. As FRTL-5 cells were repeatedly passed their morphology changed. Monolayer areas spread from beneath the follicles, and the follicles became flattened in appearance. These physical changes were coincident with dramatically increased growth rates. Early cultures (passed 3-12 times) divided on average every 49+/-1 h, whereas later passes (passes 20-25) divided every 28+/-3 h. To correlate these changes with a measure of thyroid function we assayed T(4) output. Early passage follicular cultures incubated for 6 h with sodium iodide, released on average 5.27+/- 0.33 ng/ml of T(4)/100 follicles. Later passes, or early passes treated with heptanol to down-regulate Cx32, released an average of 3.84+/-0.50 ng/ml of T(4)/100 follicles. There was a 27% difference in T(4) release between early follicular cultures, that were coupled by Cx32, and late or down-regulated early follicular cultures, that were uncoupled (P<0.0001). Collectively, the physical changes documented in this study were coincident with the loss of functional Cx32. This implies a relationship between the loss of intercellular communication and changes in morphogenic appearance, growth rate and reduced thyroid function and supports the previously postulated, tumor-suppressor role for Cx32. FRTL-5 cultures from low passage numbers are an excellent model of primary thyroid cells. However, many reports in the literature ascribe features to FRTL-5 cells that are mutually inconsistent. These differences may be resolved in the future by addressing the passage number and the conditional differences of the cultures being studied.

NASA Discipline Radiation Health↗