57Background: The stable fly, Stomoxys calcitrans, is a major blood-feeding pest of livestock that 58 has near worldwide distribution, causing an annual cost of over $2 billion for control and product 59 loss in the United States alone. Control of these flies has been limited to increased sanitary 60 management practices and insecticide application for suppressing larval stages. Few genetic 61 and molecular resources are available to help in developing novel methods for controlling stable 62 flies. 63 64 Results: This study examines stable fly biology by utilizing a combination of high-quality 65 genome sequencing, microbiome analyses, and RNA-seq analyses targeting multiple 66 developmental stages and tissues. In conjunction, manual curation of over 1600 genes was 67 used to examine gene content related to stable fly reproduction, interactions with their host, 68 host-microbe dynamics, and putative routes for control. Most notable was establishment of 69 reproduction-associated genes and identification of expanded vision, chemosensation, immune 70 repertoire, and metabolic detoxification pathway gene families. 71 72 Conclusions: The combined sequencing, assembly, and curation of the male stable fly genome 73 followed by RNA-seq and downstream analyses provide insights necessary to understand the 74 biology of this important pest. These resources and knowledge will provide the groundwork for 75 expanding the tools available to control stable fly infestations. The close relationship of 76 Stomoxys to other blood-feeding (Glossina) and non-blood-feeding flies (medflies, Drosophila, 77 house flies) will allow for understanding the evolution of blood feeding among Cyclorrhapha 78 flies. 79 80 immunity 83 Additional File 1: Supplementary Material; Additional File 2: Supplementary Tables 84 Supplementary Dataset Files: 10 85 109 or a contaminated substrate to a susceptible host, association between specific vector and 110 pathogen is not necessary) of Equine infectious anemia, African swine fever, West Nile, and Rift 111 Valley Viruses, Trypanosoma spp., and Besnoitia spp. (reviewed by [13]). The apparent low 112 vector competence of stable flies implicates the importance of immune system pathways not 113 only in regulating larval survival in microbe-rich environments but also in the inability of 114 pathogens to survive and replicate in the adult midgut following ingestion [14-16]. 115 Stable fly mate location and recognition are largely dependent upon visual cues and contact 116 pheromones [17, 18], and gravid females identify suitable oviposition sites through a 117 combination of olfactory and contact chemostimuli along with physical cues [19, 20]. Since 118 stable flies infrequently associate with their hosts, feeding only 1 to 2 times per day, on-animal 119 5 and pesticide applications are less effective control efforts than those that integrate sanitation 120 practices with fly population suppression by way of traps [21]. Given the importance of 121 chemosensory and vision pathways, repellents have been ident...

We previously reported that a peptide with the sequence Gly-Ala-Pro-Leu (GAPL) found as residues 309-312 in glycoprotein IIb alpha (GPIIb) comprises at least part of a Fg binding site on GPIIb (1). Subsequent studies demonstrated that a peptide corresponding to residues 300-312 of GPIIb alpha can bind to Fg and Vn, and is a potent inhibitor of platelet aggregation and the adhesion of activated platelets to at least four adhesive ligands: Fg, Fn, Vn, and vWf (2). Here, the production and initial characterization of polyclonal antibodies against this peptide are described. ELISAs and dot-blot assays reveal the specificity of the antibodies for the peptide immunogen. In immunoblots, the antibodies recognize GPIIb under reducing conditions. The binding of Fg to immobilized GPIIb/IIIa, the rate of clot retraction and the adhesion of stimulated platelets to Fg, fibronectin (Fn), vitronectin (Vn), and von Willebrand factor (vWf) were inhibited by these antibodies, but not by a control IgG. These results together with our earlier published data indicate that GPIIb alpha (300-312) comprises at least part of a common ligand binding site within the integrin alpha IIb subunit.

Antiplatelet agents are clinically useful as antithrombotic entities. The importance of antiplatelet agents led us to design, synthesize, and characterize a new antiplatelet peptide. This peptide is a presumptive mimic of a ligand binding site on the platelet fibrinogen receptor. Unlike peptides related to Arg-Gly-Asp-Ser and His-His-Leu-Gly-Gly-Ala-Lys-Gln-Ala-Gly-Asp-Val that bind to the fibrinogen receptor, this peptide binds to fibrinogen. The anticomplementarity hypothesis was used to design this presumptive peptide mimic of the vitronectin binding site on the fibrinogen receptor, glycoprotein IIb/IIIa complexes. The resulting peptide (Glu-His-Ile-Pro-Ala) has the characteristics of a fibrinogen binding site mimic: It binds fibrinogen and inhibits both the adhesion of platelets to fibrinogen and platelet aggregation. The peptide also inhibits the adhesion of platelets to vitronectin. The antiplatelet activity of this mimic peptide was dependent on its amino acid sequence, since closely related analogues were either inactive or less active inhibitors of platelet function than the original peptide. These results demonstrate that the peptide Glu-His-Ile-Pro-Ala has the characteristics expected of a mimic of a glycoprotein IIb/IIIa ligand binding site.