Search

Introduction: septic shock is a subset of sepsis characterized by significantly increased mortality in people. A novel proposed classification of septic shock based on 3 different phenotypes (cryptic, hyperlactatemia without persistent hypotension; vasoplegic, persistent hypotension without hyperlactatemia; and dysoxic, persistent hyperlactatemia and hypotension) seems helpful for outcome prediction. Our aim was to report the clinical features and outcome of dogs with septic shock, including the prevalence of the 3 different septic shock phenotypes, compared to dogs with uncomplicated sepsis. Methods: Dogs with sepsis, identified as SIRS plus confirmation of infection, and hospitalized in the intensive care unit were prospectively included (June 2018 –December 2019). Presence of septic shock was defined by persistent hypotension requiring vasopressor therapy or persistent hyperlactatemia after adequate fluid resuscitation Dogs with septic shock were further classified as affected by cryptic, vasoplegic, or dysoxic shock according to the previously cited human criteria. The canine Acute Patient Physiological and Laboratory Evaluation (APPLEfast) score was calculated and the presence of multiorgan dysfunction syndrome (MODS), defined as previously reported for dogs, was evaluated upon admission in the whole population. Nonparametric statistical tests were performed, and significance set at P < 0.05. Results: One-hundred-thirty-one septic dogs were enrolled; 31/131 (24%) had septic shock: cryptic (10/31, 32%), dysoxic (18/31, 58%), and vasoplegic (3/31, 10%). Septic shock dogs had significantly higher APPLEfast Scores (28, 13-38 vs 25, 6-33, P = 0.0051) and plasma lactate (3.5, 1.3-9.9 vs 2.2, 0.4-7.1, P < 0. 001), and significantly greater frequencies of MODS (74.2% vs 7%, P

NA

In this study we reported a case of co-infection with canine parvovirus (CPV) type 2a and feline panleukopenia virus (FPV) in a 3-month-old male kitten, with the presence of a parvovirus variant which is a true intermediate between CPV and FPV. The report of a viral variant which contained FPV- and CPV-specific epitopes stresses the importance of the mechanism of multistep mutation in the production of new variants and in the emergence of new viruses. This type of multistep adaptation has already been documented during the emergence of CPV and on the basis of our results, it was hypothesized that CPV had presumably started a new process of readaptation in the feline host, confirming the importance of viral host switching as a mechanism for the emergence of new viruses.

The quantification of circulating inflammatory biomarkers is relevant in the assessment of canine SIRS. The aim of this prospective study was to investigate the role of selected cytokines (IL6, IL8, IL10, IL15, MCP-1, GM-CSF; Millipore) during SIRS in dogs and their potential to differentiate sepsis from SIRS of non-infectious origin. Furthermore, a potential correlation between cytokines and other clinical and clinicopathological variables was investigated. Twenty dogs admitted to a Veterinary Teaching Hospital with a diagnosis of SIRS related to 3 different diseases (parvovirus, n=8; septic peritonitis, n=7; acute pancreatitis; n=5) were included. Ten blood donor dogs were included as healthy controls. Blood was collected upon admission by venipuncture, immediately processed at +4°C, and at least two aliquots of EDTA-plasma were stored at -80°C upon analysis. Additional laboratory analysis included measurement of C-reactive protein (CRP). Illness severity was calculated by the acute patient physiologic and laboratory evaluation fast score (APPLE). Cytokines were assayed by a multiparametric xMAP Luminex. CRP was assayed on serum using an immunoturbidimetric method on an automated chemistry analyzer. Results were analyzed by non-parametric statistic analysis. SIRS dogs have significantly higher serum CRP compared to controls (p

In recent years exosome research pubblications are rapidly expanding (1). These small vesicles (30‐100 nm) of endocytic origin are thought to participate in cell—cell communication and protein and RNA delivery. A wide range of cells have been shown to release exosomes, but they have also been detected in several biological fluids, including plasma, urine, saliva and breast milk. In particular urinary exosomes have been proposed as starting material to detect protein biomarkers of renal dysfunction and structural injury or overall to shed much insight on the health status of the kidney (2, 3, 4). We aimed to validate methods for exosome isolation from urine in a large animal model, the pig one. The swine shares with the man anatomical and physiological characteristics that make it preferred species as a pre‐clinical model, in particular for kidney functions, surgical approaches and in the view to obtain a significant amount of biological specimens (urine and blood samples, tissue for renal biopsy) compared to other animals. Exosomes were purified by differential ultracentrifugation, identified by electron microscopy and described in morphology, shape, size and distribution using atomic force microscopy (AFM). Validation methods include Western blot with pan‐exosome markers and urinary specific exosome antibodies. Exosome protein content was analyzed through nanospray liquid chromatography‐tandem mass spectrometry (LM‐MS/MS). The total exosomal RNA purificated was evaluated using gel electrophoresis and a Bioanalyzer. We found that the vesicles displayed a typical exosome‐like size and morphology as analyzed by electron microscopy and AFM. Western blot and mass spectrometry further confirmed the presence of several exosome‐associated molecules. Exosome RNA profile detected is typical of these vesicles, lacking rRNA subunits which are prominent in analysis of cellular RNA. Characterization of isolated exosomes indicates that the described isolation methods are suitable for the subsequent RNA and protein profiling.(1) Camussi G, Deregibus MC, Bruno S, Cantaluppi V, Biancone L. Exosomes/microvesicles as a mechanism of cell‐to‐cell communication. Kidney Int. 2010 Nov;78(9):838‐48. (2) Fernández‐Llama P, Khositseth S, Gonzales PA, Star RA, Pisitkun T, Knepper MA. Tamm‐Horsfall protein and urinary exosome isolation. Kidney Int. 2010 Apr;77(8):736‐42. (3) Pisitkun T, Shen RF, Knepper MA. Identification and proteomic profiling of exosomes in human urine. Proc Natl Acad Sci U S A. 2004 Sep 7;101(36):13368‐73. (4) Lässer C, Eldh M, Lötvall J. Isolation and characterization of RNA‐containing exosomes. J Vis Exp. 2012 Jan 9;(59):e3037.

Intimal hyperplasia (IH) limits the success of many vascular interventions, including bypass grafting endarterectomy and stenting. IH following vascular injury is characterized by vascular smooth muscle cells proliferation and migration, matrix deposition, inflammation. The aim of the study was to investigate, in a mild-hypercholesterolemic pig model of vascular injury: i) the role of Heme Oxygenase-1 (HO-1) in the pathogenesis of IH; ii) the molecular mechanism of prevention and therapy mediated by atorvastatin; iii) the atorvastatin-mediated induction of HO-1 in response to vascular injury. Balloon injury (BI) was used to induce IH in pig carotid arteries. Pigs were divided in 3 groups (n=7 each): standard diet; high-cholesterol diet; high-cholesterol diet and atorvastatin (80 mg/die). Carotid sections were analyzed by morphometric and immunohistochemical analyses. Hemochrome, cholesterolemia, proinflammatory cytokines profiles were also evaluated. Hypercholesterolemia induced a significant increase of total and LDL-cholesterol, exacerbated IH, increased extracellular matrix deposition, inhibited HO-1 expression and increased the local and systemic inflammatory state compared to standard diet. Atorvastatin prevented IH development by enhancing HO-1 expression, reducing the number of T-cell infiltrating the site of vascular injury, and lowering the plasmatic concentration of proinflammatory cytokines. In conclusion: i) a high-cholesterol diet induces a persistent state of vascular inflammation that worsen the vascular response following BI; ii) atorvastatin has a protective role in the pathogenesis and outcome of inflammatory response following vascular injury; iii) atorvastatin prevents IH inducing HO-1 expression in the vascular wall.