Recursive splicing is a process in which large introns are removed in multiple steps by resplicing at ratchet points - 5′ splice sites recreated after splicing1. Recursive splicing was first identified in the Drosophila Ultrabithorax (Ubx) gene1 and only three additional Drosophila genes have since been experimentally shown to undergo recursive splicing2,3. Here, we identify 197 zero nucleotide exon ratchet points in 130 introns of 115 Drosophila genes from total RNA sequencing data generated from developmental time points, dissected tissues, and cultured cells. The sequential nature of recursive splicing was confirmed by identification of lariat introns generated by splicing to and from the ratchet points. We also show that recursive splicing is a constitutive process, that depletion of U2AF inhibits recursive splicing, and that the sequence and function of ratchet points are evolutionarily conserved in Drosophila. Finally, we identified four recursively spliced human genes, one of which is also recursively spliced in Drosophila. Together these results indicate that recursive splicing is commonly used in Drosophila, occurs in human and provides insight into the mechanisms by which some large introns are removed.

Arginine and Glutamate-Rich protein 1 (ARGLU1) is a protein whose function is poorly understood, but may act in both transcription and pre-mRNA splicing. We demonstrate that the ARGLU1 gene expresses at least three distinct RNA splice isoforms – a fully spliced isoform coding for the protein, an isoform containing a retained intron that is detained in the nucleus, and an isoform containing an alternative exon that targets the transcript for nonsense mediated decay. Furthermore, ARGLU1 contains a long, highly evolutionarily conserved sequence known as an Ultraconserved Element (UCE) that is within the retained intron and overlaps the alternative exon. Manipulation of the UCE, in a reporter minigene or via random mutations in the genomic context using CRISPR/Cas9, changed the splicing pattern. Further, overexpression of the ARGLU1 protein shifted the splicing of endogenous ARGLU1 mRNA, resulting in an increase in the retained intron isoform and nonsense mediated decay susceptible isoform and a decrease in the fully spliced isoform. Taken together with data showing that functional protein knockout shifts splicing toward the fully spliced isoform, our data are consistent with a model in which unproductive splicing complexes assembled at the alternative exon lead to inefficient splicing and intron retention.

We report a head-mountable CMOS camera for recording rapid neuronal activity in freely-moving rodents using fluorescent activity reporters. This small, lightweight camera is capable of detecting small changes in light intensity (0.2% ΔI/I) at 500 fps. The camera has a resolution of 32 × 32, sensitivity of 0.62 V/lux·s, conversion gain of 0.52 μV/e- and well capacity of 2.1 Me-. The camera, containing intensity offset subtraction circuitry within the imaging chip, is part of a miniaturized epi-fluorescent microscope and represents a first generation, mobile scientific-grade, physiology imaging camera.

In this article, quantitative evaluation of optical thermographic techniques relative to the non-destructive inspection of aluminum foam material is studied. For this purpose, a set of aluminum foam specimens with flat-bottom holes (FBH) was inspected by both optical lock-in thermography (LT) and pulsed thermography (PT). Probability of detection (PoD) analysis, as a quantitative method to estimate the capability and reliability of a particular inspection technique, was studied and compared for both optical LT and PT inspection results

Advanced materials such as continuous carbon fiber-reinforced thermoplastic (CFRP) laminates are commonly used in many industries, mainly because of their strength, stiffness to weight ratio, toughness, weldability, and repairability. Structural components working in harsh environments such as satellites are permanently exposed to some sort of damage during their lifetimes. To detect and characterize these damages, non-destructive testing and evaluation techniques are essential tools, especially for composite materials. In this study, artificial intelligence was applied in combination with infrared thermography to detected and segment impact damage on curved laminates that were previously submitted to a severe thermal stress cycles and subsequent ballistic impacts. Segmentation was performed on both mid-wave and long-wave infrared sequences obtained simultaneously during pulsed thermography experiments by means of a deep neural network. A deep neural network was trained for each wavelength. Both networks generated satisfactory results. The model trained with mid-wave images achieved an F1-score of 92.74% and the model trained with long-wave images achieved an F1-score of 87.39%.

In this paper we present a fixed window level crossing sampling analog to digital convertor for bio-potential recording sensors. This is the first proposed and fully implemented fixed window level crossing ADC without local DACs and clocks. The circuit is designed to reduce data size, power, and silicon area in future wireless neurophysiological sensor systems. We built a testing system to measure bio-potential signals and used it to evaluate the performance of the circuit. The bio-potential amplifier offers a gain of 53 dB within a bandwidth of 200 Hz-20 kHz. The input-referred rms noise is 2.8 µV. In the asynchronous level crossing ADC, the minimum delta resolution is 4 mV. The input signal frequency of the ADC is up to 5 kHz. The system was fabricated using the AMI 0.5 µm CMOS process. The chip size is 1.5 mm by 1.5 mm. The power consumption of the 4-channel system from a 3.3 V supply is 118.8 µW in the static state and 501.6 µW with a 240 kS/s sampling rate. The conversion efficiency is 1.6 nJ/conversion.

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