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NASA NTRS1993

The effect of Na vapor on the Na content of chondrules

Chondrules contain higher concentrations of volatiles (Na) than expected for melt droplets in the solar nebula. Recent studies have proposed that chondrules may have formed under non-canonical nebular conditions such as in particle/gas-rich clumps. Such chondrule formation areas may have contained significant Na vapor. To test the hypothesis of whether a Na-rich vapor would minimize Na volatilization reaction rates in a chondrule analog and maintain the Na value of the melt, experiments were designed where a Na-rich vapor could be maintained around the sample. A starting material with a melting point lower that typical chondrules was required to keep the logistics of working with Na volatilization from NaCl within the realm of feasibility. The Knippa basalt, a MgO-rich alkali olivine basalt with a melting temperature of 1325 +/- 5 C and a Na2O content of 3.05 wt%, was used as the chondrule analog. Experiments were conducted in a 1 atm, gas-mixing furnace with the fO2 controlled by a CO/CO2 gas mixture and fixed at the I-W buffer curve. To determine the extent of Na loss from the sample, initial experiments were conducted at high temperatures (1300 C - 1350 C) for duration of up to 72 h without a Na-rich vapor present. Almost all (up to 98%) Na was volatilized in runs of 72 h. Subsequent trials were conducted at 1330 C for 16 h in the presence of a Na-rich vapor, supplied by a NaCl-filled crucible placed in the bottom of the furnace. Succeeding Knudsen cell weight-loss mass-spectrometry analysis of NaCl determined the P(sub Na) for these experimental conditions to be in the 10(exp -6) atm range. This value is considered high for nebula conditions but is still plausible for non-canonical environments. In these trials the Na2O content of the glass was maintained or in some cases increased; Na2O values ranged from 2.62% wt to 4.37% wt. The Na content of chondrules may be controlled by the Na vapor pressure in the chondrule formation region. Most heating events capable of producing chondrules are sufficient to volatile Na. Sodium volatilization reaction rates will be reduced to varying degrees from melt droplets, depending on the magnitude of the P(sub Na) generated. A combination of Na vapor during, and Na diffusion back into chondrules after, formation could maintain and/or enrich Na concentrations in chondrules.

Lewis, R. Dean↗

NASA NTRS2004

Microelectronic DNA assay for the detection of BRCA1 gene mutations

Mutations in BRCA1 are characterized by predisposition to breast cancer, ovarian cancer and prostate cancer as well as colon cancer. Prognosis for this cancer survival depends upon the stage at which cancer is diagnosed. Reliable and rapid mutation detection is crucial for the early diagnosis and treatment. We developed an electronic assay for the detection of a representative single nucleotide polymorphism (SNP), deletion and insertion in BRCA1 gene by the microelectronics microarray instrumentation. The assay is rapid, and it takes 30 minutes for the immobilization of target DNA samples, hybridization, washing and readout. The assay is multiplexing since it is carried out at the same temperature and buffer conditions for each step. The assay is also highly specific, as the signal-to-noise ratio is much larger than recommended value (72.86 to 321.05 vs. 5) for homozygotes genotyping, and signal ratio close to the perfect value 1 for heterozygotes genotyping (1.04).

Validation Studies↗

NASA NTRS2004

Antigenic Protein In Microgravity-Grown Human Mixed Mullerian Tumor (LN1) Cells Preserved In RNA Stabilizing Agent

Cells treated with RNAlater(TradeMark) have previously been shown to contain antigenic proteins that can be visualized using Western blot analysis. These proteins seem to be stable for several months when stored in RNA stabilizer at 4 C. Antigenic protein can be recovered from cells that have been processed using an Ambion RNAqueous(Registered TradeMark) kit to remove RNA. In this set of experiments, human mixed Mullerian tumor (LNI) cells grown on the International Space Station during Expedition 3 were examined for antigenic stability after removal of RNA. The cells were stored for three months in RNAlater(TradeMark) and RNA was extracted. The RNA filtrate containing the protein was precipitated, washed, and suspended in buffer containing sodium dodecyl sulfate (SDS). Samples containing equal concentrations of protein were loaded onto SDS-polyacrylamide gels. Proteins were separated by electrophoresis and transferred by Western blot to polyvinylidene fluoride (PVDF) membrane. The Western blots were stained with an enhanced chemiluminescent ECL(Registered Trademark) Plus detection kit (Amersham) and scanned using a Storm 840 gel image analyzer (Amersham, Molecular Dynamics). ImageQuant(Registered TradeMark) software was used to quantify the densities of the protein bands. The ground control and flight LN1 cell samples showed a similar staining pattern over time with antibodies to vimentin, glyceraldehyde-3-phosphate dehydrogenase, and epithelial membrane antigens.

Hammond, Dianne K.↗

NASA NTRS2004

Antigenic Protein In Microgravity-Grown Human Mixed Mullerian Tumor (LN1) Cells Preserved In RNA Stabilizing Agent

Cells treated with RNAlater(TradeMark) have previously been shown to contain antigenic proteins that can be visualized using Western blot analysis. These proteins seem to be stable for several months when stored in RNA stabilizer at 4 C. Antigenic protein can be recovered from cells that have been processed using an Ambion RNAqueous(Registered TradeMark) kit to remove RNA. In this set of experiments, human mixed Mullerian tumor (LN1) cells grown on the International Space Station during Expedition 3 were examined for antigenic stability after removal of RNA. The cells were stored for three months in RNAlater(TradeMark) and RNA was extracted. The RNA filtrate Containing the protein was precipitated, washed, and suspended in buffer containing sodium dodecyl sulfate (SDS). Samples containing equal concentrations of protein were loaded onto SDS-polyacrylamide gels. Proteins were separated by electrophoresis and transferred by Western blot to polyvinylidene fluoride (PVDF) membrane. The Western blots were stained with an enhanced chemiluminescent ECL(Registered TradeMark)Plus detection kit (Amersham) and scanned using a Storm 840 gel image analyzer (Amersham, Molecular Dynamics). ImageQuant(Registered TradeMark)a software was used to quantify the densities of the protein bands. The ground control and flight LN1 cell samples showed a similar staining pattern over time with antibodies to vimentin, glyceraldehyde-3-phosphate dehydrogenase, and epithelial membrane antigens.

Hammond, Dianne K.↗

NASA NTRS1988

Demixing kinetics of phase separated polymer solutions in microgravity

Phase separated solutions of two neutral polymers in buffer provide a useful and versatile medium for the partition separation of biological cells. However, the efficiency of such separations is orders of magnitude lower than the thermodynamic limit. To test the hypothesis that this inefficiency is at least partially due to the convection and sedimentation that occur during the gravity driven demixing that follows introduction of cells to the systems, a series of experiments were begun aimed at performing cell partition in a low g environment. Demixing of isopycnic three polymer solvent systems was studied, experiments were performed on KC-135 aircraft and one shuttle middeck experiment was completed. Analysis of the results of these experiments and comparisons with the predictions of scaling relations for the dependence of phase domain size on time, derived for a number of possible demixing mechanisms, are presented.

Brooks, D. E.↗

NASA NTRS1993

Technical support for digital systems technology development. Task order 1: ISP contention analysis and control

Alternatives for realizing a packet-based network switch for use on a frequency division multiple access/time division multiplexed (FDMA/TDM) geostationary communication satellite were investigated. Each of the eight downlink beams supports eight directed dwells. The design needed to accommodate multicast packets with very low probability of loss due to contention. Three switch architectures were designed and analyzed. An output-queued, shared bus system yielded a functionally simple system, utilizing a first-in, first-out (FIFO) memory per downlink dwell, but at the expense of a large total memory requirement. A shared memory architecture offered the most efficiency in memory requirements, requiring about half the memory of the shared bus design. The processing requirement for the shared-memory system adds system complexity that may offset the benefits of the smaller memory. An alternative design using a shared memory buffer per downlink beam decreases circuit complexity through a distributed design, and requires at most 1000 packets of memory more than the completely shared memory design. Modifications to the basic packet switch designs were proposed to accommodate circuit-switched traffic, which must be served on a periodic basis with minimal delay. Methods for dynamically controlling the downlink dwell lengths were developed and analyzed. These methods adapt quickly to changing traffic demands, and do not add significant complexity or cost to the satellite and ground station designs. Methods for reducing the memory requirement by not requiring the satellite to store full packets were also proposed and analyzed. In addition, optimal packet and dwell lengths were computed as functions of memory size for the three switch architectures.

Stehle, Roy H.↗

NASA NTRS1999

Information Technology and the Autonomous Control of a Mars In-Situ Propellant Production System

With the rapidly increasing performance of information technology, i.e., computer hardware and software systems, as well as networks and communication systems, a new capability is being developed that holds the clear promise of greatly increased exploration capability, along with dramatically reduced design, development, and operating costs. These new intelligent systems technologies, utilizing knowledge-based software and very high performance computer systems, will provide new design and development tools, scheduling mechanisms, and vehicle and system health monitoring capabilities. In addition, specific technologies such as neural nets will provide a degree of machine intelligence and associated autonomy which has previously been unavailable to the mission and spacecraft designer and to the system operator. One of the most promising applications of these new information technologies is to the area of in situ resource utilization. Useful resources such as oxygen, compressed carbon dioxide, water, methane, and buffer gases can be extracted and/or generated from planetary atmospheres, such as the Martian atmosphere. These products, when used for propulsion and life-support needs can provide significant savings in the launch mass and costs for both robotic and crewed missions. In the longer term the utilization of indigenous resources is an enabling technology that is vital to sustaining long duration human presence on Mars. This paper will present the concepts that are currently under investigation and development for mining the Martian atmosphere, such as temperature-swing adsorption, zirconia electrolysis etc., to create propellants and life-support materials. This description will be followed by an analysis of the information technology and control needs for the reliable and autonomous operation of such processing plants in a fault tolerant manner, as well as the approach being taken for the development of the controlling software. Finally, there will be a brief discussion of the verification and validation process so crucial to the implementation of mission-critical software.

Gross, Anthony R.↗

NASA NTRS1992

Gravity effects on cellulose assembly

The effect of microgravity on cellulose synthesis using the model system of Acetobacter xylinum was the subject of recent investigations using The National Aeronautics and Space Administration's Reduced Gravity Laboratory, a modified KC-135 aircraft designed to produce 20 sec of microgravity during the top of a parabolic dive. Approximately 40 parabolas were executed per mission, and a period of 2 x g was integral to the pullout phase of each parabola. Cellulose biosynthesis was initiated on agar surfaces, liquid growth medium, and buffered glucose during parabolic flight and terminated with 2.0% sodium azide or 50.0% ethanol. While careful ground and in-flight controls indicated normal, compact ribbons of microbial cellulose, data from five different flights consistently showed that during progression into the parabola regime, the cellulose ribbons became splayed. This observation suggests that some element of the parabola (the 20 sec microgravity phase, the 20 sec 2 x g phase, or a combination of both) was responsible for this effect. Presumably the cellulose I alpha crystalline polymorph normally is produced under strain, and the microgravity/hypergravity combination may relieve this stress to produce splayed ribbons. An in-flight video microscopy analysis of bacterial motions during a parabolic series demonstrated that the bacteria continue to synthesize cellulose during all phases of the parabolic series. Thus, the splaying may be a reflection of a more subtle alteration such as reduction of intermicrofibrillar hydrogen bonding. Long-term microgravity exposures during spaceflight will be necessary to fully understand the cellulose alterations from the short-term microgravity experiments.

manned↗

NASA NTRS1995

The Physics of Turbulence in the Boundary Layer

The geometry of the velocity field in a numerically simulated incompressible turbulent boundary layer over a flat plate at Re theta=670 has been studied using the invariants of the velocity gradient tensor. These invariants are computed at every grid point in the flow and used to form the discriminant. Of primary interest are those regions in the flow where the discriminant is positive; regions where, according to the characteristic equation, the eigenvalues of the velocity gradient tensor are complex. An observer moving with a frame of reference which is attached to a fluid particle lying within such a region would see a local flow pattern of the type stable-focus-stretching or unstable-focus-compressing. When the flow is visualized this way, continuous, connected, large-scale structures are revealed that extend from the point just below the buffer layer out to the beginning of the wake region. These structures are aligned with the mean shear close to the wall and arch in the cross-stream direction away from the wall. In some cases the structures observed are very similar to to the hairpin eddy vision of boundary layer structure proposed by Theodorsen. That the structure of the flow is revealed more effectively by the discriminant rather than by the vorticity is important and adds support to recent observations of the discriminant in a channel flow simulation. Of particular importance is the fact that the procedure does not require the use of an arbitrary threshold in the discriminant. Further analysis using computer flow visualization shows a high degree of spatial correlation between regions of positive discriminant, extreme negative pressure fluctuations and large instantaneous values of Reynolds shear stress.

Kline, Stephen↗

NASA NTRS2018

Calculating the Modal Mineralogy of Aubrite Meteorites Using X-Ray Computed Tomography

The aubrites are a group of differentiated meteorites that formed in extremely reducing conditions, with oxygen fugacities (ƒO2) ranging from ~2 to ~6 log units below the Iron-Wüstite buffer. At these extreme conditions, elements that are lithophile (silica-loving) on Earth can behave as chalcophiles (sulfur-loving) or siderophiles (metal-loving), and FeO-poor silicates and exotic sulfides can form. Elemental partitioning among minerals at these conditions are still not well understood, thus studying aubrites could help better constrain elemental behavior. Only 30 aubrites exist; however, they display various textures, which can be difficult to identify in small samples. Identifying composite clasts can help in understanding the igneous history of the aubrite parent bodies. In order to distinguish different minerals and textures in larger samples, we use X-ray computed tomography (XCT), which is a useful, non-destructive analysis that can produce a 3D representation of a meteorite's textures, structures, and modal mineralogy. This study investigates the Norton County aubrite using XCT. Norton County is a fragmental impact breccia aubrite, and was an observed fall with a main mass of 1.1 tons. This large amount of material offers an opportunity to investigate a great volume of reduced, aubritic sample and study a more diverse variety of clasts. Here we report XCT results of two samples of Norton County (NC 15417) in order to constrain the 3D modal mineralogy of silicates, sulfides, and metals, and to identify unique clasts within the matrix.

Wilbur, Zoe↗

NASA NTRS1980

Next generation communications satellites: Multiple access and network studies

Following an overview of issues involved in the choice of promising system architectures for efficient communication with multiple small inexpensive Earth stations serving hetergeneous user populations, performance evaluation via analysis and simulation for six SS/TDMA (satellite-switched/time-division multiple access) system architectures is discussed. These configurations are chosen to exemplify the essential alternatives available in system design. Although the performance evaluation analyses are of fairly general applicability, whenever possible they are considered in the context of NASA's 30/20 GHz studies. Packet switched systems are considered, with the assumption that only a part of transponder capacit is devoted to packets, the integration of circuit and packet switched traffic being reserved for further study. Three types of station access are distinguished: fixed (FA), demand (DA), and random access (RA). Similarly, switching in the satellite can be assigned on a fixed (FS) or demand (DS) basis, or replaced by a buffered store-and-forward system (SF) onboard the satellite. Since not all access/switching combinations are practical, six systems are analyzed in detail: three FS SYSTEMS, FA/FS, DA/ES, RA/FS; one DS system, DA/DS; and two SF systems, FA/SF, DA/SF. Results are presented primarily in terms of delay-throughput characteristics.

Stern, T. E.↗

NASA NTRS2008

Delay-Throughput Performance of the Deep-Space Ka-band Link

In this paper, performance of a first-in, first-out (FIFO), selective retransmission scheme for the deep-space Ka-band link is presented and compared to the performance of a comparable X-band link. In this analysis, 16 months of water vapor radiometer (WVR) and advanced water vapor radiometer (AWVR) data from the three Deep Space Network (DSN) Communication Complexes (DSCC) were used to emulate weather effects on X-band and Ka-band links from Mars. Mars Reconnaissance Orbiter (MRO) X-band and Ka-band telecommunications parameters were used for spacecraft telecommunications capabilities. One pass per week per complex was selected from MRO's Deep Space Network (DSN) schedule from April 1, 2006 to August 31, 2007 for a total of 207 passes (69 passes per complex) for this analysis. For each pass both X-band and Ka-band links were designed using at most two data rates so that the expected pass capacity would be maximized subject to a minimum availability requirement (MAR). In conjunction with the WVR/AWVR data, elevation profiles of the selected passes and models for the performance of the antennas in the DSN were used to emulate the performance of both links. It was assumed that the retransmission of the data takes place not on the same pass as the original transmission but during subsequent passes. The data collected before a pass was assumed to be a fraction of the expected capacity of the pass as calculated through the link design process. Infinite spacecraft storage was assumed to obtain an upper bound on the spacecraft storage requirement. The independent parameters of this analysis were MAR and the ratio of data collected before a pass to the expected pass capacity. Since the selected passes did not occur at regular intervals, the delay in this analysis was measured in terms of number of passes. The throughput was measured in terms of number of bits received successfully on the ground. The results indicate that reasonable delay performance could be achieved with very high throughput for relatively low MAR values for data collection to expected pass capacity ratio of around 97% for Ka-band. The results indicate that, except for very low average delay requirements, the Ka-band link provides more than twice the throughput of the X-band link for the same amount of power consumed by the spacecraft. In addition, the results indicate that the required storage onboard the spacecraft is not prohibitive and good performance could be achieved by using a buffer size less than three times the maximum amount of data collected before a pass.

Shambayati, Shervin↗

NASA NTRS2008

Delay-Throughput Performance the Deep-Space Ka-Band Link

Shambayati, Shervin↗

NASA NTRS2001

Sources, Sinks, and the Distribution of OH in the Lower Stratosphere

Extensive measurement campaigns by the NASA ER-2 research aircraft have obtained a nearly pole-to-pole database of the species that control HOx (OH + HO2) chemistry. The wide dynamic range of these in situ measurements provides an opportunity to demonstrate empirically the mechanisms that control the HOx system. Measurements in the lower stratosphere show a remarkably tight correlation of OH concentration with the solar zenith angle (SZA). This correlation is nearly invariant over latitudes ranging from 70 S to 90 N and all seasons. An analysis of the production and loss of HOx in terms of the rate determining steps of reaction sequences developed by Johnston and Podolske and Johnston and Kinnison is used to clarify the behavior of the system and to directly test our understanding of the system with observations. Calculations using in situ measurements show that the production rate of HOx is proportional to O3 and ultraviolet radiation flux. The loss rate is proportional to the concentration and the partitioning of NOy (reactive nitrogen) and the concentration of HO2. In the absence of heterogeneous reactions, the partitioning of NOy is controlled by O3 and NOx and the concentration of HO2 is controlled by NOy and O3, so that the removal rate of OH is buffered against changes in the correlation of O3 and NOy. The heterogeneous conversion of NO2 to HNO3 is not an important net source of HO, because production and removal sequences are nearly balanced. Changes in NOy partitioning resulting from heterogeneous chemistry have a large effect on the loss rates of HOx, but little or no impact on the measured abundance of OH. The enhanced loss rates at high NO2/HNO3 are offset in the data set examined here by enhanced production rates resulting from increased photolysis rates resulting from the decreased O3 column above the ER-2.

Hanisco, T. F.↗

NASA NTRS2014

Persistent Expression Changes of Fibrosis-Related Genes in the Lung Tissues of Rats Exposed to Lunar Dust Particles

The Moon's surface is covered by a layer of fine, potential reactive dust. Lunar dust contain about 1‐2% of very fine respirable dust (less than 3 micrometers). The habitable area of any lunar landing vehicle and outpost would inevitably be contaminated with lunar dust that could pose a health risk. The purpose of the study is to evaluate the toxicity of Apollo moon dust in rodents to assess the health risk of dust exposures to humans. One of the particular interests in the study is to evaluate dust‐induced changes of the expression of fibrosis‐related genes, and to identify specific signaling pathways involved in lunar dustinduced toxicity. F344 rats were exposed for 4 weeks (6h/d; 5d/wk) in nose‐only inhalation chambers to concentrations of 0 (control air), 2.1, 6.8, 21, and 61 milligrams per cubic meters of lunar dust. Five rats per group were euthanized at 1 day, 1 week, 1 month, and 3 months after the last inhalation exposure. The bronchoalveolar lavage fluid (BALF) was collected by lavaging with phosphate‐buffered saline (PBS). A zymosan‐induced luminolbased chemiluminescence assay was used to assess the activity of BAL cells. The lavaged lung tissue was snap frozen in LN2 and total RNA was isolated using the Qigen RNeasy kit. The expression of 84 fibrosisrelated genes were analyzed using the RT2 Profiler PCR Array technique. The expression of 18 genes of interest were further measured using real‐time PCR technique in all the samples. 10 out of 18 genes of interest showed persistently significant expression changes in the local lung tissue exposed to lunar dust, indicating a prolonged proinflammatory response. The expressions of several of these genes were dose‐ and time‐dependent and were significantly correlated with other pathological parameters. The potential signaling pathways and upstream regulators were further analyzed using IPA pathway analysis tool based on the gene expression data. The data presented in this study, for the first time, explore the molecular mechanisms of lunar dust induced toxicity, contributing not only the risk assessment for future space exploration, but also understandings of the dust‐induced toxicity in humans on earth.

Zhang, Ye↗

NASA NTRS1993

Analysis of cache for streaming tape drive

A tape subsystem consists of a controller and a tape drive. Tapes are used for backup, data interchange, and software distribution. The backup operation is addressed. During a backup operation, data is read from disk, processed in CPU, and then sent to tape. The processing speeds of a disk subsystem, CPU, and a tape subsystem are likely to be different. A powerful CPU can read data from a fast disk, process it, and supply the data to the tape subsystem at a faster rate than the tape subsystem can handle. On the other hand, a slow disk drive and a slow CPU may not be able to supply data fast enough to keep a tape drive busy all the time. The backup process may supply data to tape drive in bursts. Each burst may be followed by an idle period. Depending on the nature of the file distribution in the disk, the input stream to the tape subsystem may vary significantly during backup. To compensate for these differences and optimize the utilization of a tape subsystem, a cache or buffer is introduced in the tape controller. Most of the tape drives today are streaming tape drives. A streaming tape drive goes into reposition when there is no data from the controller. Once the drive goes into reposition, the controller can receive data, but it cannot supply data to the tape drive until the drive completes its reposition. A controller can also receive data from the host and send data to the tape drive at the same time. The relationship of cache size, host transfer rate, drive transfer rate, reposition, and ramp up times for optimal performance of the tape subsystem are investigated. Formulas developed will also show the advantages of cache watermarks to increase the streaming time of the tape drive, maximum loss due to insufficient cache, tradeoffs between cache and reposition times and the effectiveness of cache on a streaming tape drive due to idle times or interruptions due in host transfers. Several mathematical formulas are developed to predict the performance of the tape drive. Some examples are given illustrating the usefulness of these formulas. Finally, a summary and some conclusions are provided.

Chinnaswamy, V.↗

NASA NTRS2006

Assessing Hurricane Katrina Vegetation Damage at Stennis Space Center using IKONOS Image Classification Techniques

Hurricane Katrina inflicted widespread damage to vegetation in southwestern coastal Mississippi upon landfall on August 29, 2005. Storm damage to surface vegetation types at the NASA John C. Stennis Space Center (SSC) was mapped and quantified using IKONOS data originally acquired on September 2, 2005, and later obtained via a Department of Defense ClearView contract. NASA SSC management required an assessment of the hurricane s impact to the 125,000-acre buffer zone used to mitigate rocket engine testing noise and vibration impacts and to manage forestry and fire risk. This study employed ERDAS IMAGINE software to apply traditional classification techniques to the IKONOS data. Spectral signatures were collected from multiple ISODATA classifications of subset areas across the entire region and then appended to a master file representative of major targeted cover type conditions. The master file was subsequently used with the IKONOS data and with a maximum likelihood algorithm to produce a supervised classification later refined using GIS-based editing. The final results enabled mapped, quantitative areal estimates of hurricane-induced damage according to general surface cover type. The IKONOS classification accuracy was assessed using higher resolution aerial imagery and field survey data. In-situ data and GIS analysis indicate that the results compare well to FEMA maps of flooding extent. The IKONOS classification also mapped open areas with woody storm debris. The detection of such storm damage categories is potentially useful for government officials responsible for hurricane disaster mitigation.

Spruce, Joseph P.↗

NASA NTRS2000

Size Exclusion Chromatography Studies of the Initial Self-Association Steps of Chicken Egg White Lysozyme Nucleation

Nucleation is one of the least understood aspects of crystallogenesis. In the case of macromolecule nucleation, this understanding is further hampered by uncertainty over what precisely is being discussed. We define the process of solute self-association (aggregation, oligomerization, interaction, clustering, etc.) whereby n-mers (n > or = 2) having a crystallographic or nascent crystallographic arrangement leading to the critical nucleus reversibly form in the solution, to be part of the nucleation process. This reversible self-association process is a fundamental part of the nucleation process, and occurs as a function of the solute concentration. In the case of chicken egg white lysozyme, a considerable body of experimental evidence leads us to the conclusion that it also forms the crystal growth units. Size exclusion chromatography is a simple and direct method for determining the equilibrium constants for the self-association process. A Pharmacia FPLC system was used to provide accurate solution flow rates. The column, injection valve, and sample loop were all mounted within a temperature-controlled chamber. Chromatographically re-purified lysozyme was first dialyzed against the column equilibration buffer, with injection onto the column after several hours pre-incubation at the running temperature. Preliminary experiments, were carried out using a Toyopearl HW-50F column (1 x 50cm), equilibrated with 0.1 M sodium acetate, 5% sodium chloride, pH 4.6, at 15C. Protein concentrations from 0.1 to 4 mg/ml were employed (C(sub sat) = 1.2 mg/ml). The data from several different protein preparations consistently shows a progressively decreasing elution volume with increasing protein concentration, indicating that reversible self-association is occurring. The dotted line indicates the monomeric lysozyme elution volume. However, lysozyme interacts with the column matrix in these experiments, which complicates data analysis.Accordingly, we are testing silica-based HPLC columns in an effort to eliminate this problem and substantially reduce the column volume and experimental run time. The results and data analysis from these and subsequent experiments will be presented.

Ewing, Felecia↗