Neuropeptide hormones from the eyestalks of Jasus Lalandii

Bibliography: p. 242-264.The X-organ sinus gland complex, situated in the eyestalks of decapod crustaceans, are known to be a source of a variety of neuropeptide hormones that regulate a number of diverse physiological processes. This neuroendocrine complex was investigated in 3 crustacean species, viz. the European shore crab Carcinus maellas, and 2 spiny lobster species Jasus lalandii and Panulirus homarus by means of tissue immunocytochemistry and an enzyme-linked immunosorbent assay (ELISA). Positive immunoreactions, associated with the X-organ - sinus gland system only, were obtained with antisera raised against crustacean hyperglycaemic hormone (cHH) of the American lobster (Homarus americanus), the Mexican crayfish (Procambarus bouvieri) and the edible crab (Cancer pagurus), as well as with antisera raised against vitellogenesis-inhibiting hormone (VIH) of the H. americanus and moult-inhibiting hormone (MIH) of C. pagurus. This is the first time that the immunolocalisation of these 3 hormones have been studied in a single crustacean species. The chief results of this comparative immunocytochemical study showed that (1) neuropeptide hormones of the shore crab and the 2 spiny lobster species were sufficiently homologous in primary structure to be recognised by the heterologous antisera, thus, an indication of conserved peptide structures across the species and infraorder boundaries; (2) preabsorbed complexes of purified peptides and antisera from the edible crab did not produce any immunoreactions in tissue immunocytochemistry, nor in ELISA, thus, indicating the specificity of the anti-cHH and anti-MIH sera; (3) the anti-VIH serum demonstrated the ability to bind epitopes on cHH and MIH peptides and is, thus, not a specific antiserum in this study; (4) there is co-localisation of cHH, MIH, VIH immunoreactivity in the eyestalk neuroendocrine complexes of all 3 species studied which suggests that the different peptide hormones can be synthesized in the same neuronal cell bodies. This co-localisation of neuropeptides in the eyestalk of J lalandii was confirmed by a double-staining immunoflourescence experiment, and finally (5) immunoreactivity of antisera raised against cHH of H. americanus and MIH of C. pagurus was associated with distinct and unique peak fractions, following reverse-phase high pressure liquid chromatographic (RP-HPLC) separation of sinus gland extracts from J. lalandii. A total of 6 neuropeptide hormones belonging to the cHH/MIH/VIH peptide family were isolated, functionally characterised and sequenced from extracts of sinus glands from the South African west coast rock lobster, Jasus lalandii. This is the first complete report on these peptides from any species belonging to the Palinuridae infraorder

In Silico Screening for Pesticide Candidates against the Desert Locust Schistocerca gregaria

Adipokinetic hormone (AKH) is one of the most important metabolic neuropeptides in insects, with actions similar to glucagon in vertebrates. AKH regulates carbohydrate and fat metabolism by mobilizing trehalose and diacylglycerol into circulation from glycogen and triacylglycerol stores, respectively, in the fat body. The short peptide (8 to 10 amino acids long) exerts its function by binding to a rhodopsin-like G protein-coupled receptor located in the cell membrane of the fat body. The AKH receptor (AKHR) is, thus, a potential target for the development of novel specific (peptide) mimetics to control pest insects, such as locusts, which are feared for their prolific breeding, swarm-forming behavior and voracious appetite. Previously, we proposed a model of the interaction between the three endogenous AKHs of the desert locust, Schistocerca gregaria, and the cognate AKHR (Jackson et al., Peer J. 7, e7514, 2019). In the current study we have performed in silico screening of two databases (NCI Open 2012 library and Zinc20) to identify compounds which may fit the endogenous Schgr-AKH-II binding site on the AKHR of S. gregaria. In all, 354 compounds were found to fit the binding site with glide scores < −8. Using the glide scores and binding energies, 7 docked compounds were selected for molecular dynamic simulation in a phosphatidylcholine membrane. Of these 7 compounds, 4 had binding energies which would allow them to compete with Schgr-AKH-II for the receptor binding site and so are proposed as agonistic ligand candidates. One of the ligands, ZINC000257251537, was tested in a homospecific in vivo biological assay and found to have significant antagonistic activity

Five Neuropeptide Ligands Meet One Receptor: How Does This Tally? A Structure-Activity Relationship Study Using Adipokinetic Bioassays With the Sphingid Moth, Hippotion eson

Adipokinetic hormones (AKHs) play a major role in mobilizing stored energy metabolites during energetic demand in insects. We showed previously (i) the sphingid moth Hippotion eson synthesizes the highest number of AKHs ever recorded, viz. five, in its corpus cardiacum: two octa- (Hipes-AKH-I and II), two nona- (Hipes-AKH-III and Manse-AKH), and one decapeptide (Manse-AKH-II), which are all active in lipid mobilization (1). (ii) Lacol-AKH from a noctuid moth showed maximal AKH activity in H. eson despite sequence differences and analogs based on Lacol-AKH with modifications at positions 2, 3, 8, or at the termini, as well as C-terminally shortened analogs had reduced or no activity (2). Here we report on N-terminally shortened and modified analogs of the lead peptide, as well as single amino acid substitutions at positions 1, 4, 5, 6, and 7 by an alanine residue. Ala1 and Glu1 instead of pGlu are not tolerated well to bind to the H. eson AKH receptor, whereas Gln1 has high activity, suggesting it is endogenously cyclized. Replacing residue 5 or 7 with Ala did not alter activity much, in contrast with changes at position 4 or 6. Similarly, eliminating pGlu1, Leu2, or Thr3 from Lacol-AKH severely interfered with biological activity. This indicates that there is no core peptide sequence that can elicit the adipokinetic effect and that the overall conformation of the active peptide is required for a physiological response. AKHs achieve a biological action through binding to a receptor located on fat body cells. To date, one AKH receptor has been identified in any given insect species; we infer the same for H. eson. We aligned lepidopteran AKH receptor sequences and note that these are very similar. The results of our study is, therefore, also applicable to ligand-receptor interaction of other lepidopteran species. This information is important for the consideration of peptide mimetics to combat lepidopteran pest insects

Comparative analysis of adipokinetic hormones and their receptors in Blattodea reveals novel patterns of gene evolution

Adipokinetic hormone (AKH) is a neuropeptide produced in the insect corpora cardiaca that plays an essential role in mobilising carbohydrates and lipids from the fat body to the haemolymph. AKH acts by binding to a rhodopsin-like G protein-coupled receptor (GPCR), the adipokinetic hormone receptor (AKHR). In this study, we tackle AKH ligand and receptor gene evolution as well as the evolutionary origins of AKH gene paralogues from the order Blattodea (termites and cockroaches). Phylogenetic analyses of AKH precursor sequences point to an ancient AKH gene duplication event in the common ancestor of Blaberoidea, yielding a new group of putative decapeptides. In total, 16 different AKH peptides from 90 species were obtained. Two octapeptides and seven putatively novel decapeptides are predicted for the first time. AKH receptor sequences from 18 species, spanning solitary cockroaches and subsocial wood roaches as well as lower and higher termites, were subsequently acquired using classical molecular methods and in silico approaches employing transcriptomic data. Aligned AKHR open reading frames revealed 7 highly conserved transmembrane regions, a typical arrangement for GPCRs. Phylogenetic analyses based on AKHR sequences support accepted relationships among termite, subsocial (Cryptocercus spp.) and solitary cockroach lineages to a large extent, while putative post-translational modification sites do not greatly differ between solitary and subsocial roaches and social termites. Our study provides important information not only for AKH and AKHR functional research but also for further analyses interested in their development as potential candidates for biorational pest control agents against invasive termites and cockroaches

Discs large (Dlg1) complexes in lymphocyte activation

T cell antigen recognition involves the formation of a structured interface between antigen-presenting and T cells that facilitates the specific transmission of activating and desensitizing stimuli. The molecular machinery that organizes the signaling molecules and controls their disposition in response to activation remains poorly understood. We show here that in T cells Discs large (Dlg1), a PDZ domain-containing protein, is recruited upon activation to cortical actin and forms complexes with early participants in T cell activation. Transient overexpression of Dlg1 attenuates basal and Vav1-induced NFAT reporter activation. Reduction of Dlg1 expression by RNA interference enhances both CD3- and superantigen-mediated NFAT activation. Attenuation of antigen receptor signaling appears to be a complex, highly orchestrated event that involves the mutual segregation of important elements of the early signaling complex

Observational and Dynamical Characterization of Main-Belt Comet P/2010 R2 (La Sagra)

We present observations of comet-like main-belt object P/2010 R2 (La Sagra) obtained by Pan-STARRS 1 and the Faulkes Telescope-North on Haleakala in Hawaii, the University of Hawaii 2.2 m, Gemini-North, and Keck I telescopes on Mauna Kea, the Danish 1.54 m telescope at La Silla, and the Isaac Newton Telescope on La Palma. An antisolar dust tail is observed from August 2010 through February 2011, while a dust trail aligned with the object's orbit plane is also observed from December 2010 through August 2011. Assuming typical phase darkening behavior, P/La Sagra is seen to increase in brightness by >1 mag between August 2010 and December 2010, suggesting that dust production is ongoing over this period. These results strongly suggest that the observed activity is cometary in nature (i.e., driven by the sublimation of volatile material), and that P/La Sagra is therefore the most recent main-belt comet to be discovered. We find an approximate absolute magnitude for the nucleus of H_R=17.9+/-0.2 mag, corresponding to a nucleus radius of ~0.7 km, assuming an albedo of p=0.05. Using optical spectroscopy, we find no evidence of sublimation products (i.e., gas emission), finding an upper limit CN production rate of Q_CN<6x10^23 mol/s, from which we infer an H2O production rate of Q_H2O<10^26 mol/s. Numerical simulations indicate that P/La Sagra is dynamically stable for >100 Myr, suggesting that it is likely native to its current location and that its composition is likely representative of other objects in the same region of the main belt, though the relatively close proximity of the 13:6 mean-motion resonance with Jupiter and the (3,-2,-1) three-body mean-motion resonance with Jupiter and Saturn mean that dynamical instability on larger timescales cannot be ruled out.Comment: 23 pages, 13 figures, accepted for publication in A

Guidance on Monitoring of Marine Litter in European Seas

This publication is a Reference Report by the Joint Research Centre of the European Commission.The MSFD Technical Subgroup on Marine Litter was tasked to deliver guidance so that European Member States could initiate programmes for monitoring of Descriptor 10 of the MSFD. The present document provides the recommendations and information needed to commence the monitoring required for marine litter, including methodological protocols and categories of items to be used for the assessment of litter on the Beach, Water Column, Seafloor and Biota, including a special section on Microparticles

New genetic loci implicated in fasting glucose homeostasis and their impact on type 2 diabetes risk.

Levels of circulating glucose are tightly regulated. To identify new loci influencing glycemic traits, we performed meta-analyses of 21 genome-wide association studies informative for fasting glucose, fasting insulin and indices of beta-cell function (HOMA-B) and insulin resistance (HOMA-IR) in up to 46,186 nondiabetic participants. Follow-up of 25 loci in up to 76,558 additional subjects identified 16 loci associated with fasting glucose and HOMA-B and two loci associated with fasting insulin and HOMA-IR. These include nine loci newly associated with fasting glucose (in or near ADCY5, MADD, ADRA2A, CRY2, FADS1, GLIS3, SLC2A2, PROX1 and C2CD4B) and one influencing fasting insulin and HOMA-IR (near IGF1). We also demonstrated association of ADCY5, PROX1, GCK, GCKR and DGKB-TMEM195 with type 2 diabetes. Within these loci, likely biological candidate genes influence signal transduction, cell proliferation, development, glucose-sensing and circadian regulation. Our results demonstrate that genetic studies of glycemic traits can identify type 2 diabetes risk loci, as well as loci containing gene variants that are associated with a modest elevation in glucose levels but are not associated with overt diabetes

Pan-Cancer Analysis of lncRNA Regulation Supports Their Targeting of Cancer Genes in Each Tumor Context

Long noncoding RNAs (lncRNAs) are commonly dys-regulated in tumors, but only a handful are known toplay pathophysiological roles in cancer. We inferredlncRNAs that dysregulate cancer pathways, onco-genes, and tumor suppressors (cancer genes) bymodeling their effects on the activity of transcriptionfactors, RNA-binding proteins, and microRNAs in5,185 TCGA tumors and 1,019 ENCODE assays.Our predictions included hundreds of candidateonco- and tumor-suppressor lncRNAs (cancerlncRNAs) whose somatic alterations account for thedysregulation of dozens of cancer genes and path-ways in each of 14 tumor contexts. To demonstrateproof of concept, we showed that perturbations tar-geting OIP5-AS1 (an inferred tumor suppressor) andTUG1 and WT1-AS (inferred onco-lncRNAs) dysre-gulated cancer genes and altered proliferation ofbreast and gynecologic cancer cells. Our analysis in-dicates that, although most lncRNAs are dysregu-lated in a tumor-specific manner, some, includingOIP5-AS1, TUG1, NEAT1, MEG3, and TSIX, synergis-tically dysregulate cancer pathways in multiple tumorcontexts

Pan-cancer Alterations of the MYC Oncogene and Its Proximal Network across the Cancer Genome Atlas

Although theMYConcogene has been implicated incancer, a systematic assessment of alterations ofMYC, related transcription factors, and co-regulatoryproteins, forming the proximal MYC network (PMN),across human cancers is lacking. Using computa-tional approaches, we define genomic and proteo-mic features associated with MYC and the PMNacross the 33 cancers of The Cancer Genome Atlas.Pan-cancer, 28% of all samples had at least one ofthe MYC paralogs amplified. In contrast, the MYCantagonists MGA and MNT were the most frequentlymutated or deleted members, proposing a roleas tumor suppressors.MYCalterations were mutu-ally exclusive withPIK3CA,PTEN,APC,orBRAFalterations, suggesting that MYC is a distinct onco-genic driver. Expression analysis revealed MYC-associated pathways in tumor subtypes, such asimmune response and growth factor signaling; chro-matin, translation, and DNA replication/repair wereconserved pan-cancer. This analysis reveals insightsinto MYC biology and is a reference for biomarkersand therapeutics for cancers with alterations ofMYC or the PMN