20-HETE Mediates Ozone-Induced, Neutrophil-Independent Airway Hyper-Responsiveness in Mice

Background Ozone, a pollutant known to induce airway hyper-responsiveness (AHR), increases morbidity and mortality in patients with obstructive airway diseases and asthma. We postulate oxidized lipids mediate in vivo ozone-induced AHR in murine airways.Methodology/Principal Findings Male BALB/c mice were exposed to ozone (3 or 6 ppm) or filtered air (controls) for 2 h. Precision cut lung slices (PCLS; 250 µm thickness) containing an intrapulmonary airway (∼0.01 mm$^2$ lumen area) were prepared immediately after exposure or 16 h later. After 24 h, airways were contracted to carbachol (CCh). Log EC$_{50}$ and E$_{\text{max}}$ values were then calculated by measuring the airway lumen area with respect to baseline. In parallel studies, dexamethasone (2.5 mg/kg), or 1-aminobenzotriazol (ABT) (50 mg/kg) were given intraperitoneal injection to naïve mice 18 h prior to ozone exposure. Indomethacin (10 mg/kg) was administered 2 h prior. Cell counts, cytokine levels and liquid chromatography-mass spectrometry (LC-MS) for lipid analysis were assessed in bronchoalveolar lavage (BAL) fluid from ozone exposed and control mice. Ozone acutely induced AHR to CCh. Dexamethasone or indomethacin had little effect on the ozone-induced AHR; while, ABT, a cytochrome P450 inhibitor, markedly attenuated airway sensitivity. BAL fluid from ozone exposed animals, which did not contain an increase in neutrophils or interleukin (IL)-6 levels, increased airway sensitivity following in vitro incubation with a naïve PCLS. In parallel, significant increases in oxidized lipids were also identified using LC-MS with increases of 20-HETE that were decreased following ABT treatment.Conclusions/Significance These data show that ozone acutely induces AHR to CCh independent of inflammation and is insensitive to steroid treatment or cyclooxygenase (COX) inhibition. BAL fluid from ozone exposed mice mimicked the effects of in vivo ozone exposure that were associated with marked increases in oxidized lipids. 20-HETE plays a pivotal role in mediating acute ozone-induced AHR

Unraveling a 146 Years Old Taxonomic Puzzle: Validation of Malabar Snakehead, Species-Status and Its Relevance for Channid Systematics and Evolution

The current distribution of C. diplogramma and C. micropeltes is best explained by vicariance. The significant variation in the key taxonomic characters and the results of the molecular marker analysis points towards an allopatric speciation event or vicariant divergence from a common ancestor, which molecular data suggests to have occurred as early as 21.76 million years ago. The resurrection of C. diplogramma from the synonymy of C. micropeltes has hence been confirmed 146 years after its initial description and 134 years after it was synonymised, establishing it is an endemic species of peninsular India and prioritizing its conservation value

Dynamic Visualization System for Gaze and Dialogue Data

We report and review a visualization system capable of displaying gaze and speech data elicited from pairs of subjects interacting in a discussion. We elicit such conversation data in our first experiment, where two participants are given the task of reaching a consensus about questions involving images. We validate the system in a second experiment where the purpose is to see if a person could determine which question had elicited a certain visualization. The visualization system allows users to explore reasoning behavior and participation during multimodal dialogue interactions

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Crustacean hyperglycemic hormone family gene silencing in <i>Penaeus monodon</i> mediated through dsRNA synthesized <i>in vitro</i> from genomic and cDNA

37-43RNA interference (RNAi) is the phenomenon in which long dsRNA is able to silence cognate gene expression. In the present study, 801 bp crustacean hyperglycemic hormone 1 (CHH1) and 795 bp moult inhibiting hormone 1 (MIH1) specific dsRNAs from genomic DNA, and 316 bp gonad inhibiting hormone (GIH) specific dsRNA from cDNA were constructed in vitro. Then the specific dsRNA constructs were administered into adult shrimps (Penaeus monodon). The gene expression was studied by semi quantitative RT-PCR and by monitoring haemolymph glucose concentration, duration of moulting and expression of vitellogenin as measures of specific biological activity. The gene silencing of CHH1, MIH1 and GIH genes could be attained within 24 h of dsRNA application. GIH gene silencing was observed up to 60th h. However, a complete silencing of MIH1 and MIH2 continued to 108h post administration. Physiology of the animals injected with dsRNA of CHH1, MIH1 and GIH corroborated with the silencing of the specific genes resulting in the decrease of haemolymph glucose level, reduction in the days of moulting and expression of vitellogenin gene, respectively in adult shrimp. These results suggest the possibility of using dsRNAs of CHH family hormone genes as molecular tools for silencing inhibitory genes in turn affecting induced maturation in P. monodon.</i

Abstract Table-top Computed Lighting for Practical Digital Photography

We apply simplified image-based lighting methods to reduce the equipment, cost, time, and specialized skills required for high-quality photographic lighting of desktop-sized static objects such as museum artifacts. We place the object and a computer-steered moving-head spotlight inside a simple foam-core enclosure, and use a camera to quickly record low-resolution photos as the light scans the box interior. Optimization guided by interactive user sketching selects a small set of frames whose weighted sum best matches the target image. The system then repeats the lighting used in each of these frames, and constructs a high resolution result from re-photographed basis images. Unlike previous image-based relighting efforts, our method requires only one light source, yet can achieve high resolution light positioning to avoid multiple sharp shadows. A reduced version uses only a hand-held light, and may be suitable for battery-powered, field photography equipment that fits in a backpack. Categories and Subject Descriptors (according to ACM CCS): I.3.7 [Computer Graphics]: Three-Dimensiona

Discovery of cyclotides in the fabaceae plant family provides new insights into the cyclization, evolution, and distribution of circular proteins

Cyclotides are plant proteins whose defining struc-tural features are a head-to-tail cyclized backbone and three interlocking disulfide bonds, which in combination are known as a cyclic cystine knot. This unique structural motif confers cyclotides with exceptional resistance to proteolysis. Their endogenous function is thought to be as plant defense agents, associated with their insecticidal and larval growth-inhibitory properties. However, in addition, an array of pharmaceutically relevant biological activities has been ascribed to cyclotides, including anti-HIV, anthelmintic, uterotonic, and antimicrobial effects. So far, >150 cyclotides have been elucidated from members of the Rubiaceae, Violaceae, and Cucurbitaceae plant families, but their wider distribution among other plant families remains unclear. Clitoria ternatea (Butterfly pea) is a member of plant family Fabaceae and through its usage in traditional medicine to aid childbirth bears similarity to Oldenlandia affinis, from which many cyclotides have been isolated. Using a combination of nanospray and matrix-assisted laser desorption ionization-time-of-flight (MALDI-TOF) analyses, we examined seed extracts of C. ternatea and discovered cyclotides in the Fabaceae, the third-largest family of flowering plants. We characterized 12 novel cyclotides, thus expanding knowledge of cyclotide distribution and evolution within the plant kingdom. The discovery of cyclotides containing novel sequence motifs near the in planta cyclization site has provided new insights into cyclotide biosynthesis. In particular, MS analyses of the novel cyclotides from C. ternatea suggest that Asn to Asp variants at the cyclization site are more common than previously recognized. Moreover, this study provides impetus for the examination of other economically and agriculturally significant species within Fabaceae, now the largest plant family from which cyclotides have been described