Diet affects nearly every aspect of animal life such as development, metabolism, behavior, and aging, both directly by supplying nutrients and indirectly through gut microbiota. C. elegans feeds on bacteria, and like other animals, different bacterial diets induce distinct dietary responses in the worm. However, the lack of certain critical tools hampers the use of worms as a model for dietary signaling. Here, we genetically engineered the bacterial strain OP50, the standard laboratory diet for C. elegans, making it compatible for dsRNA production and delivery. Using this RNAi-compatible OP50 strain and the other bacterial strain HT115, we feed worms different diets while delivering RNAi to interrogate the genetic basis underlying diet-dependent differential modulation of development, metabolism, behavior, and aging. We show by RNAi that neuroendocrine and mTOR pathways are involved in mediating differential dietary responses. This genetic tool greatly facilitates the use of C. elegans as a model for dietary signaling.
With the rapid development of the industrial internet platforms, the virtualisation and ecologicalisation of manufacturing services pose new challenges to the manufacturing service capabilities, especially in terms of the improvement of publishing and transaction functions. Firstly, the demand of manufacturing service capability publishing in the market and the hierarchical structure of the industrial internet platform are analysed in this paper. Then, after defining the functional requirements, the system flow chart and the entity relationship diagram are designed. Lastly, the entire system is designed to illustrate how to publish and trade manufacturing service capabilities on the industrial internet platform.
Aberrant gene expression is a key mechanism underlying pulmonary hypertension (PH) development. The alterations of genomic chromatin accessibility and their relationship with the aberrant gene expressions in PH are poorly understood. We used bulk Assay for Transposase-Accessible Chromatin with high-throughput sequencing (ATAC-seq) and RNA sequencing (RNA-seq) in pulmonary artery smooth muscle cells (PASMCs) of chronic hypoxia-exposed rats mimicking group 3 human PH.Adult Sprague Dawley rats were commercially obtained from Hunan SJA (Hunan SJA Laboratory Animal Co., Changsha, China) and randomizedly allocated into four groups exposing to nomobaric hypoxia or normoxia for 1 or 28 days respectively. After the assessment of pulmonary hemodynamics, smooth muscle cells were isolated from intralobular arteries and simultaneously subjected to bulk Assay of ATAC-seq and RNA-seq.Hypoxic exposure for continuous 28-days, but not for 1-day, induced established PH phenotypes in rats. ATAC-seq revealed a major distribution of differential accessibility regions (DARs) annotated to the genome in out-of-promoter regions, following 1-day or 28-days hypoxia. 1188 DAR-associated genes and 378 differentially expressed genes (DEGs) were identified in rats after exposure to 1-day hypoxia, while 238 DAR-associated genes and 452 DEGs for 28-days hypoxia. Most of the DAR-associated genes or DEGs in 1-day did not overlap with that of 28-days hypoxia. A Pearson correlation analysis indicated no significant correlation between ATAC-seq and RNA-seq.The alterations in genomic chromatin accessibility and genes expression of PASMCs in the initial stage of hypoxia are distinct from the established stage of hypoxia-induced PH. The genomic differential accessibility regions may not be the main mechanisms directly underlying the differentially expressed genes observed either in the initial or established stages of PH. Thus the time-course alterations of gene expression and their possible indirect link with genomic chromatin accessibility warrant more attention in mechanistic study of pulmonary hypertension.
Abstract Mitochondrial stress within the nervous system can trigger non-cell autonomous responses in peripheral tissues. However, the specific neurons involved and their impact on organismal aging and health have remained incompletely understood. Here, we demonstrate that mitochondrial stress in γ-aminobutyric acid-producing (GABAergic) neurons in Caenorhabditis elegans ( C. elegans ) is sufficient to significantly alter organismal lifespan, stress tolerance, and reproductive capabilities. This mitochondrial stress also leads to significant changes in mitochondrial mass, energy production, and levels of reactive oxygen species (ROS). DAF-16/FoxO activity is enhanced by GABAergic neuronal mitochondrial stress and mediates the induction of these non-cell-autonomous effects. Moreover, our findings indicate that GABA signaling operates within the same pathway as mitochondrial stress in GABAergic neurons, resulting in non-cell-autonomous alterations in organismal stress tolerance and longevity. In summary, these data suggest the crucial role of GABAergic neurons in detecting mitochondrial stress and orchestrating non-cell-autonomous changes throughout the organism.
Background and Objectives: Complex congenital heart defects (CHD), which occur in approximately 3 out of 1000 neonates, require invasive open-heart surgery during the first year of life. In a recent prospective study of infants with various forms of complex CHD, 40% of neonates acquired post-operative brain injury including stroke and periventricular leukomalacia (PVL). However, studies identifying specific risk factors of post-operative stroke have been limited. Herein we examine risk factors for post-operative stroke in a mixed cohort of infants with critical CHD. Methods: Term neonates born with critical CHD from 2009 to present were identified from a single-center prospective study. Exclusion criteria included birth weight less than 2 kg, neonatal depression, perinatal seizures, and evidence of end-organ injury. Magnetic resonance imaging (MRI) to assess pre- and post-operative brain injury was performed immediately prior to and within 10 days after surgery. Results: Of 102 subjects enrolled, 85 neonates were imaged both pre- and post-operatively. Neonates were diagnosed with either hypoplastic left heart syndrome (HLHS, n=41), transposition of the great arteries (TGA, n=29) or other (n=15). A total of 9 arterial ischemic strokes were observed (10.6%), of which 7/9 were post-operative only. The incidence of stroke was significantly higher in infants with HLHS (6/41, 14.6%) than in infants with TGA (0/29, p=0.03). Time-to-surgery, head circumference, birth weight, duration of deep hypothermic circulatory arrest, and volume of pre- or post-operative PVL were not significantly correlated with incidence of stroke. Conclusion: Neonates with HLHS have a higher prevalence of post-operative embolic stroke compared to infants with TGA. Although infants with HLHS have been shown to be at a high risk for both post-operative stroke and PVL, incidence of post-operative stroke is not correlated with volume of post-operative PVL.
Transient receptor potential vanilloid (TRPV) channels are polymodal detectors of multiple environmental factors, including temperature, pH, and pressure. Inflammatory mediators enhance TRPV function through multiple signaling pathways. The lipoxygenase and epoxygenase products of arachidonic acid (AA) metabolism have been shown to directly activate TRPV1 and TRPV4, respectively. TRPV3 is a thermosensitive channel with an intermediate temperature threshold of 31-39 degrees C. We have previously shown that TRPV3 is activated by 2-aminoethoxydiphenyl borate (2APB). Here we show that AA and other unsaturated fatty acids directly potentiate 2APB-induced responses of TRPV3 expressed in HEK293 cells, Xenopus oocytes, and mouse keratinocytes. The AA-induced potentiation is observed in intracellular Ca2+ measurement, whole-cell and two-electrode voltage clamp studies, as well as single channel recordings of excised inside-out and outside-out patches. The fatty acid-induced potentiation is not blocked by inhibitors of protein kinase C and thus differs from that induced by the kinase. The potentiation does not require AA metabolism but is rather mimicked by non-metabolizable analogs of AA. These results suggest a novel mechanism regulating the TRPV3 response to inflammation, which differs from TRPV1 and TRPV4, and involves a direct action of free fatty acids on the channel.
TRPA1 forms non‐selective cation channels in polymodal nociceptors and can be activated by electrophilic prostaglandins and pungent chemicals through covalent modification. In the present study, we tested the activity of a group of non‐steroidal anti‐inflammatory drugs (NSAIDs) using electrophysiological techniques and intracellular Ca 2+ measurements. We found that specific NSAIDs can act as full agonists of TRPA1, while others are partial agonists or inactive. Extracellularly applied flufenamic, niflumic, and mefenamic acid, as well as flurbiprofen, ketoprofen, diclofenac, and indomethacin rapidly activated rat TRPA1 expressed in Xenopus oocytes and human TRPA1 in WI‐38 fibroblasts. These same NSAIDs activated human TRPA1 inducibly expressed in HEK293 cells, but the responses were absent in uninduced and parental HEK293 cells. Responses to fenamate agonists were blocked by TRPA1 antagonists, AP‐18, HC‐030031, and ruthenium red. Washout of agonist led to a rapid transition to a resting state. At subsaturating concentrations, the fenamate NSAIDs potentiated the activation of TRPA1 by AITC, cinnamaldehyde, and cold, demonstrating positive synergistic interactions. The stimulatory effect was specific to TRPA1 because flufenamic acid inhibited TRPV1, TRPV3 and TRPM8. We conclude that fenamate NSAIDs are a novel class of potent and reversible direct agonists of TRPA1.
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