Sharmila Bhattacharya

Innate Immune Responses of Drosophila melanogaster Are Altered by Spaceflight

Alterations and impairment of immune responses in humans present a health risk for space exploration missions. The molecular mechanisms underpinning innate immune defense can be confounded by the complexity of the acquired immune system of humans. Drosophila (fruit fly) innate immunity is simpler, and shares many similarities with human innate immunity at the level of molecular and genetic pathways. The goals of this study were to elucidate fundamental immune processes in Drosophila affected by spaceflight and to measure host-pathogen responses post-flight. Five containers, each containing ten female and five male fruit flies, were housed and bred on the space shuttle (average orbit altitude of 330.35 km) for 12 days and 18.5 hours. A new generation of flies was reared in microgravity. In larvae, the immune system was examined by analyzing plasmatocyte number and activity in culture. In adults, the induced immune responses were analyzed by bacterial clearance and quantitative real-time polymerase chain reaction (qPCR) of selected genes following infection with E. coli. The RNA levels of relevant immune pathway genes were determined in both larvae and adults by microarray analysis. The ability of larval plasmatocytes to phagocytose E. coli in culture was attenuated following spaceflight, and in parallel, the expression of genes involved in cell maturation was downregulated. In addition, the level of constitutive expression of pattern recognition receptors and opsonins that specifically recognize bacteria, and of lysozymes, antimicrobial peptide (AMP) pathway and immune stress genes, hallmarks of humoral immunity, were also reduced in larvae. In adults, the efficiency of bacterial clearance measured in vivo following a systemic infection with E. coli post-flight, remained robust. We show that spaceflight altered both cellular and humoral immune responses in Drosophila and that the disruption occurs at multiple interacting pathways.

Lunar Soil Simulant Uptake Produces a Concentration-Dependent Increase in Inducible Nitric Oxide Synthase Expression in Murine RAW 264.7 Macrophage Cells

One of NASAs long-term objectives is to be able to stay on the moon for extended periods, and to provide a stepping-stone for future Mars explorations. The lunar soil simulant JSC-1 has been developed by NASA from volcanic ash found in Arizona to facilitate testing of toxicity and system requirements for lunar exploration. A concentration-response study of JSC-1 was undertaken on the murine macrophage cell line RAW 264.7. Results demonstrated concentrations of 50–2000 μ g/ml JSC-1 induced enhanced expression of inducible nitric oxide synthase (iNOS). Data suggest that extraterrestrial regolith has the potential to induce an inflammatory response, and that future development of anti-inflammatory mitigative strategies may be necessary to counteract lunar dust-associated cellular toxicity.

A portable system for monitoring the behavioral activity of Drosophila.

We describe a low-cost system for monitoring the behavioral activity of the fruit fly, Drosophila melanogaster. The system is readily adaptable to one or more cameras for simultaneous recordings of behavior from different angles and can be used for monitoring multiple individuals in a population at the same time. Signal processing allows discriminating between active and inactive periods during locomotion or flying, and quantification of subtler movements related to changes in position of the wings or legs. The recordings can be taken continuously over long periods of time and can thus provide information about the dynamics of a population. The system was used to monitor responses to caffeine, changes in temperature and g-force, and activity in a variable size population.

A Miniaturized Video System for Monitoring the Locomotor Activity of Walking Drosophila Melanogaster in Space and Terrestrial Settings

A novel method is presented for monitoring movement of Drosophila melanogaster (the fruit fly) in space. Transient fly movements were captured by a

Effects of radiation and temperature on gallium nitride (GaN) metal-semiconductor-metal ultraviolet photodetectors

60, 2.5-cm-cubed monochrome video camera imaging flies illuminated by a uniform light source. The video signal from this camera was bandpass filtered (0.3-10 Hz) and amplified by an analog circuit to extract the average light changes as a function of time. The raw activity signal output of this circuit was recorded on a computer and digitally processed to extract the fly movement ldquoeventsrdquo from the waveform. These events corresponded to flies entering and leaving the image and were used for extracting activity parameters such as interevent duration. The efficacy of the system in quantifying locomotor activity was evaluated by varying environmental temperature and measuring the activity level of the flies. The results of this experiment matched those reported in the literature.

Characterization of irradiated and temperature-compensated gallium nitride surface acoustic wave resonators

The development of radiation-hardened, temperature-tolerant materials, sensors and electronics will enable lightweight space sub-systems (reduced packaging requirements) with increased operation lifetimes in extreme harsh environments such as those encountered during space exploration. Gallium nitride (GaN) is a ceramic, semiconductor material stable within high-radiation, high-temperature and chemically corrosive environments due to its wide bandgap (3.4 eV). These material properties can be leveraged for ultraviolet (UV) wavelength photodetection. In this paper, current results of GaN metal-semiconductor-metal (MSM) UV photodetectors behavior after irradiation up to 50 krad and temperatures of 15°C to 150°C is presented. These initial results indicate that GaN-based sensors can provide robust operation within extreme harsh environments. Future directions for GaN-based photodetector technology for down-hole, automotive and space exploration applications are also discussed.

Transcriptomic response of Drosophila melanogaster pupae developed in hypergravity

Conventional electronic components are prone to failure and drift when exposed to space environments, which contain harsh conditions, such as extreme variation in temperature and radiation exposure. As a result, electronic components are often shielded with heavy and complex packaging. New material platforms that leverage the radiation and temperature tolerance of wide bandgap materials can be used to develop robust electronic components without complex packaging. One such component that is vital for communication, navigation and signal processing on space exploration systems is the on-board timing reference, which is conventionally provided by a quartz crystal resonator and is prone to damage from radiation and temperature fluctuations. As a possible alternative, this paper presents the characterization of microfabricated and wide bandgap gallium nitride (GaN) surface acoustic wave (SAW) resonators in radiation environments. Ultimately, in combination with the two-dimensional gas (2DEG) layer at the AlGaN/GaN interface, high electron mobility transistor (HEMT) structures can provide a monolithic solution for timing electronics on board space systems. One-port SAW resonators are microfabricated on a GaN-on-sapphire substrate are used to explore the impact of irradiation on the device performance. The GaN-based SAW resonator was subjected to extreme temperature conditions to study the change in resonance frequency. Thermal characterization of the resonator has revealed a self-compensating property at cryogenic temperatures. In addition, GaN-on-sapphire samples were irradiated using a Cs-137 source up to 55 krads of total ionizing dose (TID). The measured frequency response and Raman spectroscopy of the GaN/sapphire SAW resonators microfabricated from the irradiated samples are presented.

Impact of gamma irradiation on GaN/sapphire surface acoustic wave resonators

Altered gravity can perturb normal development and induce corresponding changes in gene expression. Understanding this relationship between the physical environment and a biological response is important for NASAs space travel goals. We use RNA-Seq and qRT-PCR techniques to profile changes in early Drosophila melanogaster pupae exposed to chronic hypergravity (3g, or three times Earths gravity). During the pupal stage, D. melanogaster rely upon gravitational cues for proper development. Assessing gene expression changes in the pupae under altered gravity conditions helps highlight gravity-dependent genetic pathways. A robust transcriptional response was observed in hypergravity-treated pupae compared to controls, with 1513 genes showing a significant (q<0.05) difference in gene expression. Five major biological processes were affected: ion transport, redox homeostasis, immune response, proteolysis, and cuticle development. This outlines the underlying molecular and biological changes occurring in Drosophila pupae in response to hypergravity; gravity is important for many biological processes on Earth.

Estimating the speed of Drosophila locomotion using an automated behavior detection and analysis system

Space environments contain harsh conditions such as extreme temperature variations, high levels of radiation, and mechanical shocks. Resonator components made from quartz are susceptible to such environmental conditions so that complex packaging and shielding are typically required. As alternative to quartz, wide bandgap material platforms, such as gallium nitride (GaN), are being investigated due to their tolerance to harsh environments and high acoustic velocities. This paper presents the characterization of GaN/sapphire surface acoustic wave (SAW) resonators upon gamma irradiation. Microfabricated resonators were exposed to prolonged gamma ray radiation with (up to 850 krad) and the reflection characteristics (S11) were obtained before irradiation, after irradiation, and after thermal annealing at 100°C. The measured frequency response of the irradiated GaN/sapphire structures showed negligible shift in the resonance frequency (229 MHz). However, a decrease in the resonance peak intensity with increasing irradiation levels is observed. The state of the crystalline structure before and after irradiation (100 krad) is examined using rocking curve X-ray diffraction (XRD) analysis.

Protein array profiling of mouse serum, six months post whole body radiation with 56Fe

A fundamental phenotypic trait in Drosophila melanogaster is the speed of movement. Its quantification in response to environmental and experimental factors is highly useful for behavioral and neurological studies. Quantifying this behavioral characteristic in freely moving flies is difficult, and many current systems are limited to evaluating the speed of movement of one fly at a time or rely on expensive, time-consuming methods. Here, we present a novel signal processing method of quantifying the speed of multiple flies using a system with automatic behavior detection and analysis that we previously developed to quantify general activity. By evaluating the shape of the signal wave from recordings of a live and simulated single fly, a metric for speed of movement was found. The feasibility of using this metric to estimate the speed of movement in a population of flies was then confirmed by evaluating recordings taken from populations of flies maintained at two different temperatures. The results were consistent with those reported in the literature. This method provides an automated way of measuring speed of locomotion in a fly population, which will further quantify fly behavioral responses to the environment.