Antiviral RNA Interference against Orsay Virus Is neither Systemic nor Transgenerational in Caenorhabditis elegans.

UNLABELLED: Antiviral RNA-mediated silencing (RNA interference [RNAi]) acts as a powerful innate immunity defense in plants, invertebrates, and mammals. In Caenorhabditis elegans, RNAi is systemic; i.e., RNAi silencing signals can move between cells and tissues. Furthermore, RNAi effects can be inherited transgenerationally and may last for many generations. Neither the biological relevance of systemic RNAi nor transgenerational RNAi is currently understood. Here we examined the role of both pathways in the protection of C. elegans from viral infection. We studied the Orsay virus, a positive-strand RNA virus related to Nodaviridae and the first and only virus known to infect C. elegans. Immunity to Orsay virus infection requires the RNAi pathway. Surprisingly, we found that genes required for systemic or transgenerational RNAi did not have a role in antiviral defense. Furthermore, we found that Orsay virus infection did not elicit a systemic RNAi response even when a target for RNAi was provided by using transgenes. Finally, we show that viral siRNAs, the effectors of RNAi, are not inherited to a level that provides any significant resistance to viral infection in the next generation. We conclude that systemic or transgenerational RNAi does not play a role in the defense against natural Orsay virus infection. Furthermore, our data suggest that there is a qualitative difference between experimental RNAi and antiviral RNAi. Our data are consistent with a model of systemic and transgenerational RNAi that requires a nuclear or germ line component that is lacking in almost all RNA virus infections. IMPORTANCE: Since its discovery in Caenorhabditis elegans, RNAi has proven a valuable scientific tool in many organisms. In C. elegans, exogenous RNAi spreads throughout the organism and can be passed between generations; however, there has been controversy as to the endogenous role(s) that the RNAi pathway plays. One endogenous role for which spreading both within the infected organism and between generations would be advantageous is a role in viral defense. In plants, antiviral RNAi is systemic and the spread of RNAi between cells provides protection against subsequent viral infection. Here we investigated this by using the only naturally occurring virus known to infect C. elegans, Orsay virus, and surprisingly found that, in contrast to the exogenous RNAi pathway, the antiviral RNAi response targeted against this virus does not spread systemically throughout the organism and cannot be passed between generations. These results suggest that there are differences between the two pathways that remain to be discovered

Animal Counting Toolkit : a practical guide to small-boat surveys for estimating abundance of coastal marine mammals

The authors thank Synchronicity Earth, Marisla Foundation, and the US Marine Mammal Commission for seed funding for this program.Small cetaceans (dolphins and porpoises) face serious anthropogenic threats in coastal habitats. These include bycatch in fisheries; exposure to noise, plastic and chemical pollution; disturbance from boaters; and climate change. Generating reliable abundance estimates is essential to assess sustainability of bycatch in fishing gear or any other form of anthropogenic removals and to design conservation and recovery plans for endangered species. Cetacean abundance estimates are lacking from many coastal waters of many developing countries. Lack of funding and training opportunities makes it difficult to fill in data gaps. Even if international funding were found for surveys in developing countries, building local capacity would be necessary to sustain efforts over time to detect trends and monitor biodiversity loss. Large-scale, shipboard surveys can cost tens of thousands of US dollars each day. We focus on methods to generate preliminary abundance estimates from low-cost, small-boat surveys that embrace a ‘training-while-doing’ approach to fill in data gaps while simultaneously building regional capacity for data collection. Our toolkit offers practical guidance on simple design and field data collection protocols that work with small boats and small budgets, but expect analysis to involve collaboration with a quantitative ecologist or statistician. Our audience includes independent scientists, government conservation agencies, NGOs and indigenous coastal communities, with a primary focus on fisheries bycatch. We apply our Animal Counting Toolkit to a small-boat survey in Canada’s Pacific coastal waters to illustrate the key steps in collecting line transect survey data used to estimate and monitor marine mammal abundance.Publisher PDFPeer reviewe

Microfluidic Technologies for Synthetic Biology

Microfluidic technologies have shown powerful abilities for reducing cost, time, and labor, and at the same time, for increasing accuracy, throughput, and performance in the analysis of biological and biochemical samples compared with the conventional, macroscale instruments. Synthetic biology is an emerging field of biology and has drawn much attraction due to its potential to create novel, functional biological parts and systems for special purposes. Since it is believed that the development of synthetic biology can be accelerated through the use of microfluidic technology, in this review work we focus our discussion on the latest microfluidic technologies that can provide unprecedented means in synthetic biology for dynamic profiling of gene expression/regulation with high resolution, highly sensitive on-chip and off-chip detection of metabolites, and whole-cell analysis

Natural and Experimental Infection of Caenorhabditis Nematodes by Novel Viruses Related to Nodaviruses

Novel viruses have been discovered in wild Caenorahbditis nematode isolates and can now be used to explore host antiviral pathways, nematode ecology, and host-pathogen co-evolution

Dorsoventral Patterning in Hemichordates: Insights into Early Chordate Evolution

We have compared the dorsoventral development of hemichordates and chordates to deduce the organization of their common ancestor, and hence to identify the evolutionary modifications of the chordate body axis after the lineages split. In the hemichordate embryo, genes encoding bone morphogenetic proteins (Bmp) 2/4 and 5/8, as well as several genes for modulators of Bmp activity, are expressed in a thin stripe of ectoderm on one midline, historically called “dorsal.” On the opposite midline, the genes encoding Chordin and Anti-dorsalizing morphogenetic protein (Admp) are expressed. Thus, we find a Bmp-Chordin developmental axis preceding and underlying the anatomical dorsoventral axis of hemichordates, adding to the evidence from Drosophila and chordates that this axis may be at least as ancient as the first bilateral animals. Numerous genes encoding transcription factors and signaling ligands are expressed in the three germ layers of hemichordate embryos in distinct dorsoventral domains, such as pox neuro, pituitary homeobox, distalless, and tbx2/3 on the Bmp side and netrin, mnx, mox, and single-minded on the Chordin-Admp side. When we expose the embryo to excess Bmp protein, or when we deplete endogenous Bmp by small interfering RNA injections, these expression domains expand or contract, reflecting their activation or repression by Bmp, and the embryos develop as dorsalized or ventralized limit forms. Dorsoventral patterning is independent of anterior/posterior patterning, as in Drosophila but not chordates. Unlike both chordates and Drosophila, neural gene expression in hemichordates is not repressed by high Bmp levels, consistent with their development of a diffuse rather than centralized nervous system. We suggest that the common ancestor of hemichordates and chordates did not use its Bmp-Chordin axis to segregate epidermal and neural ectoderm but to pattern many other dorsoventral aspects of the germ layers, including neural cell fates within a diffuse nervous system. Accordingly, centralization was added in the chordate line by neural-epidermal segregation, mediated by the pre-existing Bmp-Chordin axis. Finally, since hemichordates develop the mouth on the non-Bmp side, like arthropods but opposite to chordates, the mouth and Bmp-Chordin axis may have rearranged in the chordate line, one relative to the other

Competing conservation objectives for predators and prey: estimating killer whale prey requirements for chinook salmon

Ecosystem-based management (EBM) of marine resources attempts to conserve interacting species. In contrast to single-species fisheries management, EBM aims to identify and resolve conflicting objectives for different species. Such a conflict may be emerging in the northeastern Pacific for southern resident killer whales (Orcinus orca) and their primary prey, Chinook salmon (Oncorhynchus tshawytscha). Both species have at-risk conservation status and transboundary (Canada–US) ranges. We modeled individual killer whale prey requirements from feeding and growth records of captive killer whales and morphometric data from historic live-capture fishery and whaling records worldwide. The models, combined with caloric value of salmon, and demographic and diet data for wild killer whales, allow us to predict salmon quantities needed to maintain and recover this killer whale population, which numbered 87 individuals in 2009. Our analyses provide new information on cost of lactation and new parameter estimates for other killer whale populations globally. Prey requirements of southern resident killer whales are difficul

Additional file 1: of A soluble truncated tau species related to cognitive dysfunction and caspase-2 is elevated in the brain of Huntington’s disease patients

Figure S1. Levels of tau-5-reactive soluble proteins revealed by direct Western blotting are lowered in HD patients than non-HD individuals. Figure S2. Levels of tau-13-immunoreactive Δtau314 proteins, following normalization to levels of tau-5-reactive proteins (T-tau (WB)) revealed by direct Western blotting, are higher in HD patients than non-HD individuals. Figure S3. Levels of glyceraldehyde 3-phosphate dehydrogenase (GAPDH) are comparable between HD patients and non-HD individuals of the large cohort. Figure S4. The relationships of Δtau314 protein levels with demographic characteristics of subjects. Table S1. Demographic and neuropathological characteristics of human subjects. Table S2. Comparison of demographic characteristics of HD patients and non-HD individuals from the HUB-ICO-IDIBELL Biobank, Spain used in the study of proteins in the prefrontal cortex (BA8). Table S3. A statistical comparison of protein levels of HD patients and non-HD individuals from the HUB-ICO-IDIBELL Biobank, Spain. Table S4. A statistical comparison of levels of proteins revealed by direct Western blotting (WB) probed with tau-5 antibody in the prefrontal cortex (BA8/9) of HD patients and non-HD individuals from the NIH NeuroBioBank and the New York Brain Bank. Table S5. A statistical comparison of protein levels of proteins revealed by direct Western blotting (WB) probed with tau-5 antibody in the caudate nucleus of HD patients and non-HD individuals from the NIH NeuroBioBank. File S1 Supplementary references. (ZIP 8370 kb

Targeted peripheral focused ultrasound stimulation attenuates obesity-induced metabolic and inflammatory dysfunctions

AbstractObesity, a growing health concern, is associated with an increased risk of morbidity and mortality. Chronic low-grade inflammation is implicated in obesity-driven metabolic complications. Peripheral focused ultrasound stimulation (pFUS) is an emerging non-invasive technology that modulates inflammation. Here, we reasoned that focused ultrasound stimulation of the liver may alleviate obesity-related inflammation and other comorbidities. After 8 weeks on a high-fat high-carbohydrate “Western” diet, C57BL/6J mice were subjected to either sham stimulation or focused ultrasound stimulation at the porta hepatis. Daily liver-focused ultrasound stimulation for 8 weeks significantly decreased body weight, circulating lipids and mitigated dysregulation of adipokines. In addition, liver-focused ultrasound stimulation significantly reduced hepatic cytokine levels and leukocyte infiltration. Our findings demonstrate the efficacy of hepatic focused ultrasound for alleviating obesity and obesity-associated complications in mice. These findings suggest a previously unrecognized potential of hepatic focused ultrasound as a possible novel noninvasive approach in the context of obesity. </jats:p