Response of a Li-glass/multi-anode photomultiplier detector to collimated thermal-neutron beams

The response of a position-sensitive Li-glass scintillator detector being developed for thermal-neutron detection with 6 mm position resolution has been investigated using collimated beams of thermal neutrons. The detector was moved perpendicularly through the neutron beams in 0.5 to 1.0 mm horizontal and vertical steps. Scintillation was detected in an 8 X 8 pixel multi-anode photomultiplier tube on an event-by-event basis. In general, several pixels registered large signals at each neutron-beam location. The number of pixels registering signal above a set threshold was investigated, with the maximization of the single-hit efficiency over the largest possible area of the detector as the primary goal. At a threshold of ~50% of the mean of the full-deposition peak, ~80% of the events were registered in a single pixel, resulting in an effective position resolution of ~5 mm in X and Y. Lower thresholds generally resulted in events demonstrating higher pixel multiplicities, but these events could also be localized with ~5 mm position resolution.Comment: 23 pages, 8 figure

Light-yield response of liquid scintillators using 2–6 MeV tagged neutrons

Knowledge of the neutron light-yield response is crucial to the understanding of scintillator-based neutron detectors. In this work, neutrons from 2–6MeV have been used to study the scintillation light-yield response of the liquid scintillators NE 213A, EJ 305, EJ 331 and EJ 321P using event-by-event waveform digitization. Energy calibration was performed using a GEANT4 model to locate the edge positions of the Compton distributions produced by gamma-ray sources. The simulated light yield for neutrons from a PuBe source was compared to measured recoil proton distributions, where neutron energy was selected by time-of-flight. This resulted in an energy-dependent Birks parameterization to characterize the non-linear response to the lower energy neutrons. The NE 213A and EJ 305 results agree very well with existing data and are reproduced nicely by the simulation. New results for EJ 331 and EJ 321P, where the simulation also reproduces the data well, are presented

Response of a Li-glass/multi-anode photomultiplier detector to α-particles from ²⁴¹Am

The response of a position-sensitive Li-glass scintillator detector to -particles from a collimated ²⁴¹Am source scanned across the face of the detector has been measured. Scintillation light was read out by an 8 x 8 pixel multi-anode photomultiplier and the signal amplitude for each pixel has been recorded for every position on a scan. The pixel signal is strongly dependent on position and in general several pixels will register a signal (a hit) above a given threshold. The effect of this threshold on hit multiplicity is studied, with a view to optimize the single-hit efficiency of the detector

Formation and Toxicity of Soluble Polyglutamine Oligomers in Living Cells

Aggregation and cytotoxicity of mutant proteins containing an expanded number of polyglutamine (polyQ) repeats is a hallmark of several diseases, including Huntington's disease (HD). Within cells, mutant Huntingtin (mHtt) and other polyglutamine expansion mutant proteins exist as monomers, soluble oligomers, and insoluble inclusion bodies (IBs). Determining which of these forms constitute a toxic species has proven difficult. Recent studies support a role for IBs as a cellular coping mechanism to sequester levels of potentially toxic soluble monomeric and oligomeric species of mHtt.When fused to a fluorescent reporter (GFP) and expressed in cells, the soluble monomeric and oligomeric polyglutamine species are visually indistinguishable. Here, we describe two complementary biophysical fluorescence microscopy techniques to directly detect soluble polyglutamine oligomers (using Htt exon 1 or Htt(ex1)) and monitor their fates in live cells. Photobleaching analyses revealed a significant reduction in the mobilities of mHtt(ex1) variants consistent with their incorporation into soluble microcomplexes. Similarly, when fused to split-GFP constructs, both wildtype and mHtt(ex1) formed oligomers, as evidenced by the formation of a fluorescent reporter. Only the mHtt(ex1) split-GFP oligomers assembled into IBs. Both FRAP and split-GFP approaches confirmed the ability of mHtt(ex1) to bind and incorporate wildtype Htt into soluble oligomers. We exploited the irreversible binding of split-GFP fragments to forcibly increase levels of soluble oligomeric mHtt(ex1). A corresponding increase in the rate of IBs formation and the number formed was observed. Importantly, higher levels of soluble mHtt(ex1) oligomers significantly correlated with increased mutant cytotoxicity, independent of the presence of IBs.Our study describes powerful and sensitive tools for investigating soluble oligomeric forms of expanded polyglutamine proteins, and their impact on cell viability. Moreover, these methods should be applicable for the detection of soluble oligomers of a wide variety of aggregation prone proteins

Retinal Biosynthesis in Fungi: Characterization of the Carotenoid Oxygenase CarX from Fusarium fujikuroi

The car gene cluster of the ascomycete Fusarium fujikuroi encodes two enzymes responsible for torulene biosynthesis (CarRA and CarB), an opsin-like protein (CarO), and a putative carotenoid cleaving enzyme (CarX). It was presumed that CarX catalyzes the formation of the major carotenoid in F. fujikuroi, neurosporaxanthin, a cleavage product of torulene. However, targeted deletion of carX did not impede neurosporaxanthin biosynthesis. On the contrary, ΔcarX mutants showed a significant increase in the total carotenoid content, indicating an involvement of CarX in the regulation of the pathway. In this work, we investigated the enzymatic activity of CarX. The expression of the enzyme in β-carotene-accumulating Escherichia coli cells led to the formation of the opsin chromophore retinal. The identity of the product was proven by high-performance liquid chromatography and gas chromatography-mass spectrometry. Subsequent in vitro assays with heterologously expressed and purified CarX confirmed its β-carotene-cleaving activity and revealed its capability to produce retinal also from other substrates, such as γ-carotene, torulene, and β-apo-8′-carotenal. Our data indicate that the occurrence of at least one β-ionone ring in the substrate is required for the cleavage reaction and that the cleavage site is determined by the distance to the β-ionone ring. CarX represents the first retinal-synthesizing enzyme reported in the fungal kingdom so far. It seems likely that the formed retinal is involved in the regulation of the carotenoid biosynthetic pathway via a negative feedback mechanism