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High Speed Intensified Video Observations of TLEs in Support of PhOCAL

The third observing season of PhOCAL (Physical Origins of Coupling to the upper Atmosphere by Lightning) was conducted over the U.S. High Plains during the late spring and summer of 2013. The goal was to capture using an intensified high-speed camera, a transient luminous event (TLE), especially a sprite, as well as its parent cloud-to-ground (SP+CG) lightning discharge, preferably within the domain of a 3-D lightning mapping array (LMA). The co-capture of sprite and its SP+CG was achieved within useful range of an interferometer operating near Rapid City. Other high-speed sprite video sequences were captured above the West Texas LMA. On several occasions the large mesoscale convective complexes (MCSs) producing the TLE-class lightning were also generating vertically propagating convectively generated gravity waves (CGGWs) at the mesopause which were easily visible using NIR-sensitive color cameras. These were captured concurrent with sprites. These observations were follow-ons to a case on 15 April 2012 in which CGGWs were also imaged by the new Day/Night Band on the Suomi NPP satellite system. The relationship between the CGGW and sprite initiation are being investigated. The past year was notable for a large number of elve+halo+sprite sequences sequences generated by the same parent CG. And on several occasions there appear to be prominent banded modulations of the elves' luminosity imaged at >3000 ips. These stripes appear coincident with the banded CGGW structure, and presumably its density variations. Several elves and a sprite from negative CGs were also noted. New color imaging systems have been tested and found capable of capturing sprites. Two cases of sprites with an aurora as a backdrop were also recorded. High speed imaging was also provided in support of the UPLIGHTS program near Rapid City, SD and the USAFA SPRITES II airborne campaign over the Great Plains.

Lyons, Walter A.↗

In-Space Crew-Collaborative Task Scheduling

For all past and current human space missions, the final scheduling of tasks to be done in space has been devoid of crew control, flexibility, and insight. Ground controllers, with minimal input from the crew, schedule the tasks and uplink the timeline to the crew or uplink the command sequences to the hardware. Prior to the International Space Station (ISS), the crew could make requests about tomorrow s timeline, they could omit a task, or they could request that something in the timeline be delayed. This lack of control over one's own schedule has had negative consequences. There is anecdotal consensus among astronauts that control over their own schedules will mitigate the stresses of long duration missions. On ISS, a modicum of crew control is provided by the job jar. Ground controllers prepare a task list (a.k.a. "job jar") of non-conflicting tasks from which jobs can be chosen by the in space crew. Because there is little free time and few interesting non-conflicting activities, the task-list approach provides little relief from the tedium of being micro-managed by the timeline. Scheduling for space missions is a complex and laborious undertaking which usually requires a large cadre of trained specialists and suites of complex software tools. It is a giant leap from today s ground prepared timeline (with a job jar) to full crew control of the timeline. However, technological advances, currently in-work or proposed, make it reasonable to consider scheduling a collaborative effort by the ground-based teams and the in-space crew. Collaboration would allow the crew to make minor adjustments, add tasks according to their preferences, understand the reasons for the placement of tasks on the timeline, and provide them a sense of control. In foreseeable but extraordinary situations, such as a quick response to anomalies and extended or unexpected loss of signal, the crew should have the autonomous ability to make appropriate modifications to the timeline, extend the timeline, or even start over with a new timeline. The Vision for Space Exploration (VSE), currently being pursued by the National Aeronautics and Space Administration (NASA), will send humans to Mars in a few decades. Stresses on the human mind will be exacerbated by the longer durations and greater distances, and it will be imperative to implement stress-reducing innovations such as giving the crew control of their daily activities.

Jaap, John↗

Lunar mare volcanism: Mixing of distinct, mantle source regions with KREEP-like component

Mare basalts comprise less than 1% of the lunar crust, but they constitute our primary source of information on the moon's upper mantle. Compositional variations between mare basalt suites reflect variations in the mineralogical and geochemical composition of the lunar mantle which formed during early lunar differentiation (4.5-4.4 AE). Three broad suites of mare basalt are recognized: very low-Ti (VLT) basalts with TiO2 less than 1 wt%, low-Ti basalts with TiO2 = 2-4 wt%, and high-Ti basalts with TiO2 = 10-14 wt%. Important subgroups include the Apollo 12 ilmenite basalts (TiO2 = 5-6 wt%), aluminous low-Ti mare basalts (TiO2 = 2-4 wt%, Al2O3 = 10-14 wt%), and the newly discovered Very High potassium (VHK) aluminous low-Ti basalts, with K2O = 0.4-1.5 wt%. The mare basalt source region has geochemical characteristics complementary to the highlands crust and is generally thought to consist of mafic cumulates from the magma ocean which formed the felsic crust by feldspar flotation. The progressive enrichment of mare basalts in Fe/Mg, alkalis, and incompatible trace elements in the sequence VLT basalt yields low-Ti basalt yields high-Ti basalt is explained by the remelting of mafic cumulates formed at progressively shallower depths in the evolving magma ocean. This model is also consistent with the observed decrease in compatible element concentrations and the progressive increase in negative Eu anomalies.

Shervais, John W.↗

Thrust Measurement of Dielectric Barrier Discharge (DBD) Plasma Actuators: New Anti-Thrust Hypothesis, Frequency Sweeps Methodology, Humidity and Enclosure Effects

We discuss thrust measurements of Dielectric Barrier Discharge (DBD) plasma actuators devices used for aerodynamic active flow control. After a review of our experience with conventional thrust measurement and significant non-repeatability of the results, we devised a suspended actuator test setup, and now present a methodology of thrust measurements with decreased uncertainty. The methodology consists of frequency scans at constant voltages. The procedure consists of increasing the frequency in a step-wise fashion from several Hz to the maximum frequency of several kHz, followed by frequency decrease back down to the start frequency of several Hz. This sequence is performed first at the highest voltage of interest, then repeated at lower voltages. The data in the descending frequency direction is more consistent and selected for reporting. Sample results show strong dependence of thrust on humidity which also affects the consistency and fluctuations of the measurements. We also observed negative values of thrust, or "anti-thrust", at low frequencies between 4 Hz and up to 64 Hz. The anti-thrust is proportional to the mean-squared voltage and is frequency independent. Departures from the parabolic anti-thrust curve are correlated with appearance of visible plasma discharges. We propose the anti-thrust hypothesis. It states that the measured thrust is a sum of plasma thrust and anti-thrust, and assumes that the anti-thrust exists at all frequencies and voltages. The anti-thrust depends on actuator geometry and materials and on the test installation. It enables the separation of the plasma thrust from the measured total thrust. This approach enables more meaningful comparisons between actuators at different installations and laboratories. The dependence on test installation was validated by surrounding the actuator with a grounded large-diameter metal sleeve. Strong dependence on humidity is also shown; the thrust significantly increased with decreasing humidity, e.g., 44 percent increase as relative humidity changed from 18 percent and dew point 33 degF to 50 percent and dew point of 57 degF.

Ashpis, David E.↗

Dielectric Barrier Discharge (DBD) Plasma Actuators Thrust-Measurement Methodology Incorporating New Anti-Thrust Hypothesis

We discuss thrust measurements of Dielectric Barrier Discharge (DBD) plasma actuators devices used for aerodynamic active flow control. After a review of our experience with conventional thrust measurement and significant non-repeatability of the results, we devised a suspended actuator test setup, and now present a methodology of thrust measurements with decreased uncertainty. The methodology consists of frequency scans at constant voltages. The procedure consists of increasing the frequency in a step-wise fashion from several Hz to the maximum frequency of several kHz, followed by frequency decrease back down to the start frequency of several Hz. This sequence is performed first at the highest voltage of interest, then repeated at lower voltages. The data in the descending frequency direction is more consistent and selected for reporting. Sample results show strong dependence of thrust on humidity which also affects the consistency and fluctuations of the measurements. We also observed negative values of thrust or "anti-thrust", at low frequencies between 4 Hz and up to 64 Hz. The anti-thrust is proportional to the mean-squared voltage and is frequency independent. Departures from the parabolic anti-thrust curve are correlated with appearance of visible plasma discharges. We propose the anti-thrust hypothesis. It states that the measured thrust is a sum of plasma thrust and anti-thrust, and assumes that the anti-thrust exists at all frequencies and voltages. The anti-thrust depends on actuator geometry and materials and on the test installation. It enables the separation of the plasma thrust from the measured total thrust. This approach enables more meaningful comparisons between actuators at different installations and laboratories. The dependence on test installation was validated by surrounding the actuator with a large diameter, grounded, metal sleeve.

Ashpis, David E.↗

The Plasma Interaction Experiment (PIX) description and test program

The plasma interaction experiment (PIX) is a battery powered preprogrammed auxiliary payload on the LANDSAT-C launch. This experiment is part of a larger program to investigate space plasma interactions with spacecraft surfaces and components. The varying plasma densities encountered during available telemetry coverage periods are deemed sufficient to determine first order interactions between the space plasma environment and the biased experimental surfaces. The specific objectives of the PIX flight experiment are to measure the plasma coupling current and the negative voltage breakdown characteristics of a solar array segment and a gold plated steel disk. Measurements will be made over a range of surface voltages up to plus or minus kilovolt. The orbital environment will provide a range of plasma densities. The experimental surfaces will be voltage biased in a preprogrammed step sequence to optimize the data returned for each plasma region and for the available telemetry coverage.

Ignaczak, L. R.↗

Trichococcus Patagoniensis sp. nov., a Facultative Anaerobe that grows at -5 C, Isolated from Penguin Guano in Chilean Patagonia

A novel, extremely psychrotolerant, facultative anaerobe, strain PmagGl(sup T), was isolated from guano of Magellanic penguins (Spheniscus magellanicus) collected in Chilean Patagonia. Gram-variable, motile cocci with a diameter of 1.3-2.0 micrometers were observed singularly or in pairs, short chains and irregular conglomerates. Growth occurred within the pH range 6.0-10.0, with optimum growth at pH 8.5. The temperature range for growth of the novel isolate was from -5 to 35 C, with optimum growth at 28-30 C. Strain PmagG1(sup T) did not require NaCl, as growth was observed in the presence of 0-6.5% NaCl with optimum growth at 0.5% (w/v). Strain PmagGl(sup T) was a catalase-negative chemo-organoheterotroph that used sugars and some organic acids as substrates. The metabolic end products were lactate, formate, acetate, ethanol and Con. Strain PmagG1(sup T) was sensitive to ampicillin, tetracycline, chloramphenicol, rifampicin, kanamycin and gentamicin. The G+C content of its genomic DNA was 45.8 mol%. 16S rRNA gene sequence analysis showed 100 % similarity of strain PmagG1(sup T) with Trichococcus collinsii ATCC BAA-296(sup T), but DNA-DNA hybridization between them demonstrated relatedness values of less than 45 plus or minus 1%. Another phylogenetically closely related species, Trichococcus pasteurii, showed 99.85 % similarity by 16s rRNA sequencing and DNA-DNA hybridization showed relatedness values of 47 plus or minus 1.5%. Based on genotypic and phenotypic characteristics, the novel species Trichococcus patagoniensis sp. nov. is proposed, with strain PmagG1(sup T) (=ATCC BAA-756(sup T)=JCM 12176(sup T)=CIP 108035(sup T)) as the type strain.

Pikuta, Elena V.↗

The Plasma Interaction Experiment /PIX/ - Description and flight qualification test program

The Plasma Interaction Experiment (PIX) is a battery powered preprogrammed auxiliary payload on the Landsat-C launch. This experiment is part of a larger program to investigate space plasma interactions with spacecraft surfaces and components. The varying plasma densities encountered during available telemetry coverage periods are deemed sufficient to determine first order interactions between the space plasma environment and the biased experimental surfaces. The specific objectives of the PIX flight experiment are to measure the plasma coupling current and the negative voltage breakdown characteristics of a solar array segment and a gold plated steel disk. Measurements will be made over a range of surface voltages up to plus or minus 1 kilovolt. The orbital environment will provide a range of plasma densities. The experimental surfaces will be voltage-biased in a preprogrammed step sequence to optimize the data returned for each plasma region and for the available telemetry coverage.

Ignaczak, L. R.↗

Spectral evolution of young stellar objects

An evolutionary sequence, from protostars to pre-main sequence stars, for the classification of young stellar objects is derived by comparing the predictions of the theoretical protostar models of Adams and Shu (AS, 1986) with the morphological classification scheme of Lada and Wilking (1984). It is shown that the AS models adequately explain the emergent spectral energy distributions of unidentified objects with negative spectral indices in the mid-IR and near-IR in both Taurus and Ophiuchus. If the infalling dust envelope is then completely removed, the spectra of the underlying stars and nebular disks used by AS provide a natural explanation for the near-IR and mid-IR excesses and the positive spectral indices of embedded T Tauri stars. It is found that the addition of a simple physical model for residual dust envelopes can reproduce the far-IR excesses found in some of these T Tauri stars.

Adams, Fred C.↗

Haloanaerobium kushneri sp. nov., an obligately halophilic, anaerobic bacterium from an oil brine

Three strains, designated VS-751T, VS-511 and VS-732, of a strictly anaerobic, moderately halophilic, Gram-negative, rod-shaped bacterium were isolated from a highly saline (15-20%) brine from an oil reservoir in central Oklahoma, USA. The optimal concentration of NaCl for growth of these three strains was 2 M (12%), and the strains also grew in the presence of an additional 1 M MgCl2. The strains were mesophilic and grew at a pH range of 6-8. Carbohydrates used by all three strains included glucose, fructose, arabinose, galactose, maltose, mannose, cellobiose, sucrose and inulin. Glucose fermentation products included ethanol, acetate, H2 and CO2, with formate produced by two of the three strains. Differences were noted among strains in the optimal temperature and pH for growth, the maximum and minimum NaCl concentration that supported growth, substrate utilization and cellular fatty acid composition. Despite the phenotypic differences among the three strains, analysis of the 16S rRNA gene sequences and DNA-DNA hybridizations showed that these three strains were members of the same genospecies which belonged to the genus Haloanaerobium. The phenotypic and genotypic characteristics of strains VS-751T, VS-511 and VS-732 are different from those of previously described species of Haloanaerobium. It is proposed that strain VS-751T (ATCC 700103T) be established as the type strain of a new species, Haloanaerobium kushneri.

NASA Discipline Exobiology↗

Spectral evolution of young stellar objects

An evolutionary sequence, from protostars to pre-main-sequence stars, is suggested for the classification of young stellar objects. This sequence is derived by comparing the predictions of the theoretical models of Adams and Shu with the morphological classification scheme of Lada and Wilking. We first define the spectral index in the near- and mid-infrared, n is identical to d log(nu F sub nu)/d log nu, and then interpret the class of sources with negative spectral indices as protostars. The inferred mass infall rates for these objects are generally consistent with the measured gas temperatures of approx. 35 K in Ophiuchus, and of approx. 10 K in Taurus. Fitting the data requires us to adopt cloud rotation rates in Ophiuchus which are typically an order of magnitude greater than in Taurus, and we speculate on the mechanistic origin for this difference. Next, we consider a subclass of T Tauri stars with near- and mid-infrared excesses and positive or zero spectral indices. We find that the objects with the steeper indices can be understood as the post-infall products from the collapse of rotating cloud cores, where the infrared excesses arise from the simple reprocessing of visible stellar photons in optically thick but spatially thin disks. The sources with flatter spectra may require massive accretion disks. Given the existence of protostars and naked star/disk systems, there is a natural interpretation of another subclass of T Tauri stars, those with two peaks in their emergent spectral energy distributions. These are readily explained as intermediate cases in which dust envelopes still surround the stars and disks. Finally, we find that the theory can be extended to explain the spectral energy distribution of FU Orionis, a famous outburst source. Our model suggests that FU Orionis has a disk, but it offers no discrimination between the competing ideas that the outburst took place on the star or in the disk.

Adams, F. C.↗

Specialized physiological studies in support of manned space flight

Subjects were tested for tolerance to lower body negative pressure (LBNP) before and after acute dehydration by working intermittenly for two hours without fluid replacement. On the second day there-after the LBNP tests were repeated before and after acute dehydration. The LBNP test consisted of 5 min long consective stages at -20, -30, -40, -50 and -60 Torr. Tests were terminated when syncope was imminent or the full sequence was completed. Tolerance was expressed in terms of cumulative stress in Torr x min. Measurements of body mass, density, fat fraction and total body water (TBW) were made before and after acclimation. Blood volume and its constituents were determined before and after each of the four LBNP tests. During LBNP, heart rate, blood pressure, and changes in calf and forearm volume were recorded every minute. Results showed: acute dehydration caused a significant loss in average LBNP tolerance on all subjects. Acclimation to heat did not significantly affect LBNP tolerance in hydrated subjects but significantly improved it on dehydrated subjects.

Luft, U. C.↗

A UAV-Based Fog Collector Design for Fine-Scale Aerobiological Sampling

Airborne microbes are found throughout the troposphere and into the stratosphere. Knowing how the activity of airborne microorganisms can alter water, carbon, and other geochemical cycles is vital to a full understanding of local and global ecosystems. Just as on the land or in the ocean, atmospheric regions vary in habitability; the underlying geochemical, climatic, and ecological dynamics must be characterized at different scales to be effectively modeled. Most aerobiological studies have focused on a high level: 'How high are airborne microbes found?' and 'How far can they travel?' Most fog and cloud water studies collect from stationary ground stations (point) or along flight transects (1D). To complement and provide context for this data, we have designed a UAV-based modified fog and cloud water collector to retrieve 4D-resolved samples for biological and chemical analysis.Our design uses a passive impacting collector hanging from a rigid rod suspended between two multi-rotor UAVs. The suspension design reduces the effect of turbulence and potential for contamination from the UAV downwash. The UAVs are currently modeled in a leader-follower configuration, taking advantage of recent advances in modular UAVs, UAV swarming, and flight planning.The collector itself is a hydrophobic mesh. Materials including Tyvek, PTFE, nylon, and polypropylene monofilament fabricated via laser cutting, CNC knife, or 3D printing were characterized for droplet collection efficiency using a benchtop atomizer and particle counter. Because the meshes can be easily and inexpensively fabricated, a set can be pre-sterilized and brought to the field for 'hot swapping' to decrease cross-contamination between flight sessions or use as negative controls.An onboard sensor and logging system records the time and location of each sample; when combined with flight tracking data, the samples can be resolved into a 4D volumetric map of the fog bank. Collected samples can be returned to the lab for a variety of analyses. Based on a review of existing flight studies, we have identified ion chromatography, metagenomic sequencing, cell staining and quantification, and ATP quantification as high-priority assays for implementation. Support for specific toxicology assays, such as methylmercury quantification, is also planned.

Design↗

A UAV-Based Fog Collector Design for Fine-Scale Aerobiological Sampling

Airborne microbes are found throughout the troposphere and into the stratosphere. Knowing how the activity of airborne microorganisms can alter water, carbon, and other geochemical cycles is vital to a full understanding of local and global ecosystems. Just as on the land or in the ocean, atmospheric regions vary in habitability; the underlying geochemical, climatic, and ecological dynamics must be characterized at different scales to be effectively modeled. Most aerobiological studies have focused on a high level: 'How high are airborne microbes found?' and 'How far can they travel?' Most fog and cloud water studies collect from stationary ground stations (point) or along flight transects (1D). To complement and provide context for this data, we have designed a UAV-based modified fog and cloud water collector to retrieve 4D-resolved samples for biological and chemical analysis.Our design uses a passive impacting collector hanging from a rigid rod suspended between two multi-rotor UAVs (Unmanned Aerial Vehicles). The suspension design reduces the effect of turbulence and potential for contamination from the UAV downwash. The UAVs are currently modeled in a leader-follower configuration, taking advantage of recent advances in modular UAVs, UAV swarming, and flight planning.The collector itself is a hydrophobic mesh. Materials including Tyvek, PTFE, nylon, and polypropylene monofilament fabricated via laser cutting, CNC knife, or 3D printing were characterized for droplet collection efficiency using a benchtop atomizer and particle counter. Because the meshes can be easily and inexpensively fabricated, a set can be pre-sterilized and brought to the field for 'hot swapping' to decrease cross-contamination between flight sessions or use as negative controls.An onboard sensor and logging system records the time and location of each sample; when combined with flight tracking data, the samples can be resolved into a 4D volumetric map of the fog bank. Collected samples can be returned to the lab for a variety of analyses. Based on a review of existing flight studies, we have identified ion chromatography, metagenomic sequencing, cell staining and quantification, and ATP quantification as high-priority assays for implementation. Support for specific toxicology assays, such as methyl-mercury quantification, is also planned.

Design↗

PRIME: A bottom-up approach to probabilistic rule development

PRIME is a system to be used by an intelligent machine to allow it to operate in an abstract but uncertain (or stochastic) environment. It maintains a model of the effects of the machine's actions in the form of a rule base, which is induced from experience. This bottom-up approach to rule development allows the model to adapt to changes in the environment. Each rule consists of a condition under which the rule is active, an action, the effect of the action on the environment, and an estimate of the probability of this effect occurring. The effect probabilities are used to model the uncertainty in the environment, permitting multiple possible effects for a single action under a particular set of conditions. The objective of the intelligent machine is to satisfy user-specified goals with maximum probability of success. PRIME fulfills this requirement in two ways: it continuously updates the rule base with the most recent information, to ensure the validity of the model; and it generates plans which have the maximum probability of achieving the goals, based on the probability estimates in the rule base. PRIME is composed of three main processes: exploration, generalization, and planning. In exploration, the machine executes various randomly chosen actions, observes the effects on the environment, and updates the rule base accordingly. This process is used to develop the rule base in simulation, as well as to supplement the current knowledge during normal operation. Generalization is the procedure used to induce general rules from experience, which is encoded in the form of specific rules. These general rules extend the machine's knowledge to situations which have not been encountered yet, thereby increasing the capability of the machine to plan effectively. Planning is the process of constructing an optimal sequence of actions to satisfy a goal, using the rule base to predict the effects of these actions and to determine the probability of success of the plan. The rule representation and many other data structures were specifically chosen to maximize the efficiency of these processes. A simulated environment was designed to test the performance of PRIME. The results of experimentation were largely negative. The main problem was that the domain coverage of the rules was inadequate for the number of rules stored in the rule base, due to redundancies in general rules and numerous rules covering ineffective actions. It was determined that a more efficient generalization, and some form of goal-directed exploration, are necessary in order to solve most of the current deficiencies in PRIME.

Miller, Scott A.↗

Gelidivirgula Patagoniensis Gen. Nov., Sp. Nov., A Novel Psychrotolerant, Sporeforming Anaerobe Isolated from Magellanic Penguin Guano in Patagonia, Chile

A novel obligately anaerobic, psychrotrophic bacterium, strain PPP2(sup T), was isolated from guano of the Magellanic penguin (Spheniscus magellanicus) in Patagonia, Chile. The Gram-positive, sporeforming, straight rods with sizes 0.6-0.9 x 3.0-5.0 microns, are motile by peritrichous flagella. Growth was observed to occur within the pH range 6.0-9.5 (optimum pH x), and temperature range 2-28 C (optimum 20 C). The novel isolate does not require NaCl for growth, but is halotolerant and growth was observed between 0 and 7 % NaCl (w/v) with optimum at 0.5 % (w/v). The new isolate is a catalase negative chemoorganohetherotroph with fermentative metabolism and uses as substrates: peptone, Bacto-tryptone, Casamino acids, and yeast extract. The major metabolic products are: acetate, butyrate, ethanol, and hydrogen is a minor gas product.. Strain PPP2 was sensitive to ampicillin, tetracycline, chloramphenicol, rifampin, kanamycin, and gentamycin. The G+C content of the DNA is 43.6 mol%. On the basis of 16S rDNA gene sequences and phenotypic characteristics, it is proposed that the strain PPP2(sup T) (= ATCC BAA-755(sup T) = JSM ...(sup T)) is assigned to the new genus Gelidivirgula gen. nov., as a representative of the new species, Gelidivirgula patagonensis sp. nov.

Pikuta, Elena V.↗

A search for diffuse interstellar bands in the ultraviolet

The International Ultraviolet Explorer (IUE) Satellite was used to collect interstellar extinction curves for 58 moderately reddened lines of sight. Spectral types of the background stars ranged from O7 to A0. All luminosity classes are represented, but early B main-sequence stars predominate. E(B-V) reddening values ranged from 0.14 to 1.13 mag, with an average of 0.63 mag. These extinction curves, less some 10 curves judged of lower quality, were co-added in order to reduce the random and fixed-pattern noise of the instrument. The resulting averaged curve was examined for evidence of small-scale structure, with negative results. The 2-sigma limits were found to be 0.09 mag for wavelengths between 1150 A and 2000 A, and 0.16 mag for wavelengths between 2000 A and 2300 A. Because of mismatch errors in weak stellar lines, it is unlikely that much improvement in these results is possible. The implications of these negative results are discussed with particular reference to the MgO theory of interstellar grains and diffuse bands.

Seab, C. G.↗

Zuotin, a putative Z-DNA binding protein in Saccharomyces cerevisiae

A putative Z-DNA binding protein, named zuotin, was purified from a yeast nuclear extract by means of a Z-DNA binding assay using [32P]poly(dG-m5dC) and [32P]oligo(dG-Br5dC)22 in the presence of B-DNA competitor. Poly(dG-Br5dC) in the Z-form competed well for the binding of a zuotin containing fraction, but salmon sperm DNA, poly(dG-dC) and poly(dA-dT) were not effective. Negatively supercoiled plasmid pUC19 did not compete, whereas an otherwise identical plasmid pUC19(CG), which contained a (dG-dC)7 segment in the Z-form was an excellent competitor. A Southwestern blot using [32P]poly(dG-m5dC) as a probe in the presence of MgCl2 identified a protein having a molecular weight of 51 kDa. The 51 kDa zuotin was partially sequenced at the N-terminal and the gene, ZUO1, was cloned, sequenced and expressed in Escherichia coli; the expressed zuotin showed similar Z-DNA binding activity, but with lower affinity than zuotin that had been partially purified from yeast. Zuotin was deduced to have a number of potential phosphorylation sites including two CDC28 (homologous to the human and Schizosaccharomyces pombe cdc2) phosphorylation sites. The hexapeptide motif KYHPDK was found in zuotin as well as in several yeast proteins, DnaJ of E.coli, csp29 and csp32 proteins of Drosophila and the small t and large T antigens of the polyoma virus. A 60 amino acid segment of zuotin has similarity to several histone H1 sequences. Disruption of ZUO1 in yeast resulted in a slow growth phenotype.

NASA Discipline Exobiology↗