USE OF SOLENSIA (FRUNEVETMAB) IN NONDOMESTIC FELIDS (PANTHERA TIGRIS, LYNX RUFUS, PANTHERA UNCIA) GUIDED BY PROTEIN SEQUENCE ANALYSIS.

Osteoarthritis and degenerative joint disease affect many species of nondomestic felid and negatively impact quality of life in managed care settings. Previously, pain management options were limited because of the frequency of comorbidities such as renal disease and common difficulty encountered in medication administration. SolensiaTM (frunevetmab) is a felinized monoclonal antibody that binds to nerve growth factor in arthritic joints, thereby inhibiting pain response cascades, and produces marked improvement in clinical signs and quality of life in domestic cats diagnosed with osteoarthritis. Protein sequence analysis was performed to inform the application of frunevetmab in four individuals from three nondomestic felid species (Panthera tigris, Lynx rufus, and Panthera uncia) diagnosed with degenerative joint disease to predict safety, contraindications, and likely response to treatment. Patients were then treated utilizing doses extrapolated from domestic cats and following manufacturer guidelines for administration, and clinical response was evaluated over a minimum 2-mon period. Significant improvement was noted in clinical signs in all four animals, resulting in marked improvement in mobility, lameness, activity level, demeanor, and overall quality of life. Frunevetmab presents an excellent adjunctive therapeutic alternative for nondomestic felids suffering from degenerative joint disease and may complement or decrease the need for other conventional therapeutic regimens.

Leishmania (L.) amazonensis LaLRR17 increases parasite entry in macrophage by a mechanism dependent on GRP78.

Leishmaniases affect 12 million people worldwide. They are caused by Leishmania spp., protozoan parasites transmitted to mammals by female phlebotomine flies. During the life cycle, promastigote forms of the parasite live in the gut of infected sandflies and convert into amastigotes inside the vertebrate macrophages. The parasite evades macrophage's microbicidal responses due to virulence factors that affect parasite phagocytosis, survival and/or proliferation. The interaction between Leishmania and macrophage molecules is essential to phagocytosis and parasite survival. Proteins containing leucine-rich repeats (LRRs) are common in several organisms, and these motifs are usually involved in protein­protein interactions. We have identified the LRR17 gene, which encodes a protein with 6 LRR domains, in the genomes of several Leishmania species. We show here that promastigotes of Leishmania (L.) amazonensis overexpressing LaLRR17 are more infective in vitro. We produced recombinant LaLRR17 protein and identified macrophage 78 kDa glucose-regulated protein (GRP78) as a ligand for LaLRR17 employing affinity chromatography followed by mass spectrometry. We showed that GRP78 binds to LaLRR17 and that its blocking precludes the increase of infection conferred by LaLRR17. Our results are the first to report LRR17 gene and protein, and we hope they stimulate further studies on how this protein increases phagocytosis of Leishmania.

Fever as an evolutionary agent to select immune complexes interfaces.

We herein analyzed all available protein-protein interfaces of the immune complexes from the Protein Data Bank whose antigens belong to pathogens or cancers that are modulated by fever in mammalian hosts. We also included, for comparison, protein interfaces from immune complexes that are not significantly modulated by the fever response. We highlight the distribution of amino acids at these viral, bacterial, protozoan and cancer epitopes, and at their corresponding paratopes that belong strictly to monoclonal antibodies. We identify the "hotspots", i.e. residues that are highly connected at such interfaces, and assess the structural, kinetic and thermodynamic parameters responsible for complex formation. We argue for an evolutionary pressure for the types of residues at these protein interfaces that may explain the role of fever as a selective force for optimizing antibody binding to antigens.

Equine simplified acute physiology score: Personalised medicine for the equine emergency patient.

BACKGROUND: Scoring models are useful tools that guide the attending clinician in gauging the severity of disease evolution and in evaluating the efficacy of treatment. There are few tools available with this purpose for the non-human patient, including horses. We aimed (i) to adapt the simplified acute physiology score 3 (SAPS-3) model for the equine species, reaching a margin of accuracy greater than 75% in the calculation of the probability of survival/death and (ii) to build a decision tree that helps the attending veterinarian in assessment of the clinical evolution of the equine patient. METHODS: From an initial pool of 5568 medical records from University-based Veterinary Hospitals, a final cohort of 1000 was further mined manually for data extraction. A set of 19 variables were evaluated and tested by five machine learning data mining algorithms. RESULTS: The final scoring model, named EqSAPS for equine simplified acute physiology score, reached 91.83% of correct estimates (post hoc) for probability of death within 24 hours upon hospitalization. The area under receiver operating characteristic curve for outcome 'death' was 0.742, while for 'survival' was 0.652. The final decision tree was able to refine prognosis of patients whose EqSAPS score suggested 'death'. CONCLUSION: EqSAPS is a useful tool to gauge the severity of the clinical presentation of the equine patient.

p38{alpha} MAP kinase controls IL-17 synthesis in vogt-koyanagi-harada syndrome and experimental autoimmune uveitis.

PURPOSE. Interleukin (IL)-17, which is responsible for the initial influx of leukocytes into the target tissue, was recently described as the main cytokine involved in autoimmune diseases. Vogt-Koyanagi-Harada (VKH) syndrome is a significant cause of noninfectious blindness in the world. Herein the authors aimed at unraveling the involvement of IL-17 in VKH and in experimental autoimmune uveitis, focusing on the signaling pathways involved in IL-17 synthesis. METHODS. Mice were immunized with 161-180 peptide and pertussis toxin. Draining lymph node cells, harvested 21 days after immunization, were cultured in the presence or absence of p38alpha mitogen-activated protein kinase (MAPK) inhibitor (SB203580) and assayed for cytokine production and quantification of CD4(+)IL-17(+) cells. Mice received intraocular injections of SB203580, and disease severity was evaluated by histologic examination of the enucleated eyes at day 21. CD4(+) lymphocytes from MSK-1/2-deficient mice, human CD4(+) cells silenced with MSK1 siRNA, or peripheral blood mononuclear cells (PBMCs) from VKH patients were cultured in the presence or absence of p38alpha MAPK inhibitor and then assayed for IL-17, IFN-gamma, and IL-4 production. RESULTS. The inhibition of p38alpha MAPK fully blocked the synthesis of IL-17 by PBMCs from VKH patients and lymphocytes from EAU mice. The absence of the msk1/2 gene resulted in failure to produce IL-17 by murine and human lymphocytes. Interestingly, intraocular injections of SB203580 in EAU mice did not suppress development of the disease. CONCLUSIONS. These data show that p38alpha MAPK-MSK1/2 is involved in the control of IL-17 synthesis by CD4(+) T cells and that inhibition of p38alpha MAPK in vitro suppresses IL-17 synthesis but that inhibition of this kinase in vivo did not protect from EAU.