Biomarker di sepsi e disfunzione d'organo nel cane e nel gatto

Background Multiorgan dysfunction syndrome (MODS), the progressive dysfunction of organ systems following an acute threat to systemic homeostasis, is a common complication of systemic inflammatory response syndrome (SIRS) and sepsis in the intensive care unit (ICU). Bloodstream biomarkers for MODS prediction and prognostication have received growing attention in human medicine. Literature concerning MODS occurrence and significance is scant in dogs and absent in cats. Criteria for the systematic evaluation of organ dysfunction are lacking, and the use of diagnostic and prognostic biomarkers is limited in veterinary critical care medicine. Aims The aim of the proposed research is to investigate novel biomarkers for the prediction of illness severity, organ dysfunction and prognosis in critical dogs and cats hospitalized in the ICU of a veterinary university hospital (VUH). Materials and Methods Critically ill dogs and cats hospitalized in the ICU of the VUH of Bologna during the PhD (2014-2017) and diagnosed with SIRS and sepsis have been selected for the studies. Conclusion The role of biomarkers is becoming crucial in critical care medicine, as they can assist in patient management and predict early and late complications of critical illness. The present thesis contributes to characterize SIRS and sepsis in dogs and cats, and gives novel insights on biomarkers of disease severity and organ dysfunction. A systematic screening for MODS has been proposed in the performed studies, highlighting the need to early recognize this condition at the time of ICU admission and during hospital stay. The prognostic impact of selected organ dysfunction and MODS development has been observed. The presented results improve our understanding of the host response to inflammation and infection, and are the basis for an on-going process to characterize MODS and its sequelae in critical care veterinary medicine

Diagnosis of sepsis in dogs by measuring carbonylated proteins (PCOs) and paraoxonase (PON-1)

An early diagnosis of sepsis could allow a better prognosis and avoid the abuse of antibiotic administration; unfortunately, in veterinary medicine, specific and sensitive markers of sepsis are not available.Because Protein Carbonyls (PCOs), that result from protein oxidation, are widely used in human medicine as sepsis markers , the aim of our study was to validate an ELISA kit (Enzolifesciences, 3V Chimica, Roma) on canine serum and to measure PCOs, after a preliminary validation study, in three groups (homogeneous for age and size): healthy dogs without clinical or laboratory abnormalities (A, n=14), dogs with septic (B, n=14) and non-septic inflammation (C, n=12) at the first presentation and without previous treatments. Moreover, Paraoxonase-1, a negative acute phase protein with anti-oxidant properties (PON-1) was measured in each group with a method already validated in dogs.A Kruskal-Wallis test followed by a Wilcoxon signed rank test was used to evaluate differences between groups.The ELISA method for measuring PCOs showed a very good precision (coefficient of variation <12%) and a good accuracy in spiking-recovery tests.Compared with controls, the concentration of PCOs was significantly higher either in dogs with sepsis (P<0.001) or in dogs with non-septic inflammation (P=0.005) but no significant differences were found between the two groups of sick dogs. Conversely, PON-1 was significantly lower in sick dogs compared with controls (P<0.001 for both groups) and in septic dogs compared with dogs with non-septic inflammation (P=0.001).A negative correlation between the two markers was found (P<0.001, r=-0.594) Receiver operating characteristic (ROC) curves demonstrated that both markers may discriminate dogs with sepsis with the other groups. However, PCO was less sensitive than PON-1 in diagnosing sepsis.Future studies should be focused on the association of PCOs with other inflammatory markers, as well as the possible prognostic role of PCOs based on the outcome of the enrolled patients

Classification of Septic Shock Phenotypes Based on the Presence of Hypotension and Hyperlactatemia in Cats

open8noBackground: Three different phenotypes of septic shock based on changes in blood pressure and lactate are recognized in people. Dysoxic shock, representing the combination of fluid-refractory hypotension and hyperlactatemia, is characterized by greater disease severity and mortality compared to cryptic shock (hyperlactatemia alone) and vasoplegic shock (hypotension with normal blood lactate). Little is known about septic shock and specifically its phenotypes in cats. Objective: To analyze the characteristics and prognostic implications of three septic shock phenotypes in cats with sepsis. Methods: Cats with septic shock were prospectively included. Septic shock was defined by the presence of hypotension (mean blood pressure <60 mmHg) requiring vasopressor support and/or persistent hyperlactatemia (>4 mmol/L) and classified in three subgroups: dysoxic shock, vasoplegic shock and cryptic shock. Clinical and clinicopathological variables including APPLEfast and APPLEfull scores, occurrence of multi-organ dysfunction syndrome (MODS; presence of at least two dysfunctional organs simultaneously) and outcome were compared among subgroups. Cats with sepsis showing normal blood pressure and lactate concentrations hospitalized during the study period were included as uncomplicated sepsis, and compared to cats with septic shock for selected variables. Length of hospital stay and mortality were evaluated in the whole study population. Odds ratios for mortality were calculated using logistic regression analysis. Significance was set at P < 0.05. Results: The study enrolled 48 cats with uncomplicated sepsis and 37 cats with septic shock (dysoxic shock n = 17; vasoplegic shock n = 11; cryptic shock n = 7). Cats with dysoxic shock had significantly higher APPLEfast and APPLEfull scores compared to vasoplegic and cryptic shock. Mortality rates were not significantly different among cryptic (57%), dysoxic (65%) and vasoplegic shock (91%), while MODS occurrence was significantly lower in cats with cryptic shock (57%) compared to patients affected by dysoxic (94%) and vasoplegic (100%) shock. Cats with septic shock had higher frequency of MODS and greater mortality rate than cats with uncomplicated sepsis. Conclusion: Despite similar in-hospital mortality, cats with dysoxic and vasoplegic shock are characterized by having higher occurrence of multi- organ dysfunction compared to cats affected by cryptic shock. Results from this study suggest novel means of identifying high-risk subgroups of septic cats.openTroia R.; Buzzurra F.; Ciuffoli E.; Mascalzoni G.; Foglia A.; Magagnoli I.; Dondi F.; Giunti M.Troia R.; Buzzurra F.; Ciuffoli E.; Mascalzoni G.; Foglia A.; Magagnoli I.; Dondi F.; Giunti M

Concomitant Infections With Canine Parvovirus Type 2 and Intracellular Tick-Borne Pathogens in Two Puppy Dogs

In this report the concomitant infection with canine parvovirus type 2 (CPV-2), Hepatozoon canis and Ehrlichia canis in two puppy dogs from Southern Italy is described. Dogs were referred to a veterinary university hospital for the acute onset of lethargy and gastrointestinal signs. A complete clinical and clinicopathological evaluation was carried out and the multiple infection was confirmed by microscopic detection of inclusion bodies in peripheral blood smear, rapid immunoenzymatic tests, indirect fluorescent antibody tests, and molecular assays. Sequence analysis revealed that the CPV-2 identified belonged to the 2c variant and had amino acid residues in the predicted VP2 protein typical of “Asian-like” strains widespread in Asia and occasionally reported in Romania, Nigeria and Italy, particularly in the region of Sicily. Numerous monocytes were infected by both H. canis gamonts and E. canis morulae, suggesting that this co-infection is not accidental and that E. canis preferably infects those cells parasitized by H. canis. The clinical presentation of these animals was severe but supportive cares associated with early etiological therapy allowed a good prognosis. Movement of puppies from geographic areas where vector-borne pathogens are endemic must be carefully evaluated and core vaccinations and ectoparasite prevention treatments must be rigorously adopted

Renal adenocarcinoma-associated erythrocytosis in a cat: Clinicopathological features and immunohistochemical results

This report documents a case of secondary inappropriate erythrocytosis in a cat with renal cell adenocarcinoma, its stabilization through manual erythrocytapheresis, and the EPO-immunostaining on the affected kidney. An 11-year-old cat was presented with lethargy, weight loss and polyuria/polydipsia. An abdominal mass was detected upon physical examination. Clinicopathological work-up revealed marked erythrocytosis (HCT value 64.8%), renal azotemia and decreased urine specific gravity (USG). An abdominal ultrasound was performed, localizing the mass in the right kidney. Serum erythropoietin (EPO) was above the reference interval (RI), and the cytology of the mass was indicative of renal carcinoma. Manual erythrocytapheresis was performed in order to stabilize the patient before surgery, improving the cat’s clinical and clinicopathological condition. After nephrectomy, EPO and creatinine concentrations returned within the RI, while the USG markedly increased. Histopathology confirmed the diagnosis of renal adenocarcinoma. Immunohistochemistry with anti-EPO antibody revealed diffuse and strong cytoplasmatic positivity in tumor cells

2H,3H-decafluoropentane-based nanodroplets: New perspectives for oxygen delivery to hypoxic cutaneous tissues

Perfluoropentane (PFP)-based oxygen-loaded nanobubbles (OLNBs) were previously proposed as adjuvant therapeutic tools for pathologies of different etiology sharing hypoxia as a common feature, including cancer, infection, and autoimmunity. Here we introduce a new platform of oxygen nanocarriers, based on 2H,3H-decafluoropentane (DFP) as core fluorocarbon. These new nanocarriers have been named oxygen-loaded nanodroplets (OLNDs) since DFP is liquid at body temperature, unlike gaseous PFP. Dextran-shelled OLNDs, available either in liquid or gel formulations, display spherical morphology, ~600 nm diameters, anionic charge, good oxygen carrying capacity, and no toxic effects on human keratinocytes after cell internalization. In vitro OLNDs result more effective in releasing oxygen to hypoxic environments than former OLNBs, as demonstrated by analysis through oxymetry. In vivo, OLNDs effectively enhance oxy-hemoglobin levels, as emerged from investigation by photoacoustic imaging. Interestingly, ultrasound (US) treatment further improves transdermal oxygen release from OLNDs. Taken together, these data suggest that US-activated, DFP-based OLNDs might be innovative, suitable and cost-effective devices to topically treat hypoxia-associated pathologies of the cutaneous tissues

Plasma procalcitonin concentrations predict organ dysfunction and outcome in dogs with sepsis

Abstract Background Procalcitonin (PCT) is a valuable prognostic biomarker in human sepsis that is predictive of organ dysfunction, septic shock and mortality. Data on PCT in dogs is limited. This study aimed to investigate the prognostic value of baseline and serial PCT measurements in dogs with sepsis and to determine the association between PCT and sepsis severity and the presence of organ dysfunction. PCT concentrations were measured in citrated plasma samples collected from 53 dogs with sepsis at the time of admission (T0, n = 53) and at 24 h (T1, n = 35) and 48 h (T2, n = 30) post-admission using a commercial ELISA. Dogs were classified by sepsis severity (sepsis without organ dysfunction; severe sepsis; septic shock) and outcome (survivors; non-survivors). Organ dysfunctions were recorded at T0 and during hospitalization, and the APPLEfast score calculated at T0. Healthy dogs (n = 12) were used as controls. Results There were 18 septic dogs without organ dysfunction, 24 dogs with severe sepsis and 11 with septic shock. Baseline PCT concentrations were significantly greater in dogs with sepsis compared to healthy controls (P < 0.0001), and in dogs with septic shock compared to dogs without cardiovascular compromise (P = 0.01). Baseline PCT was significantly correlated with organ dysfunction (P = 0.003). Declining PCT concentrations were documented in survivors at T1 and T2 compared to PCT at T0 (P = 0.0006), and PCT clearance at 24 h was significantly higher in survivors (n = 38) compared to non-survivors (n = 15) (P = 0.037). Canine APPLEfast score was not predictive of sepsis severity, the development of MODS or outcome. Conclusion In dogs with sepsis, PCT concentrations at hospital admissions are predictive of organ dysfunction and septic shock. Serial procalcitonin monitoring may offer valuable prognostic information in canine sepsis, wherein early decreases in PCT concentrations are associated with survival

Cell-Free DNA, High-Mobility Group Box-1, and Procalcitonin Concentrations in Dogs With Gastric Dilatation–Volvulus Syndrome

Canine gastric dilatation–volvulus (GDV) is a life-threatening disease characterized by extensive tissue ischemia, tissue hypoperfusion, and systemic inflammation. Biomarkers that better reflect the severity of gastric necrosis and systemic inflammation would aid clinicians in the management of these patients. This study aimed to investigate the prognostic significance of cell-free DNA (cfDNA), high-mobility group box-1 (HMGB1), and procalcitonin (PCT) in dogs with GDV. Concentrations of cfDNA, HMGB1, and PCT were measured in citrated plasma samples collected from 29 dogs with GDV at hospital admission. Additional data collected included baseline lactate concentrations, APPLEfast score, evidence of gastric necrosis, occurrence of postoperative complications, and outcome. Twenty-four healthy dogs were sampled as controls. Continuous variables between groups were compared with the Mann–Whitney U and correlations between continuous variables were assessed by calculation of Spearman’s correlation coefficient. Alpha was set at 0.05. Dogs with GDV had significantly greater concentrations of cfDNA, HMGB1, and PCT compared to controls (P = 0.0009, P = 0.004, and P = 0.009, respectively). PCT concentrations were significantly higher in non-survivors compared to survivors (P = 0.008). Dogs with gastric necrosis had significantly greater lactate concentrations compared to dogs without gastric necrosis (P = 0.0005). The APPLEfast score was not prognostic. Lactate and PCT concentrations were moderately, positively correlated (rs 0.51, P = 0.0005). Concentrations of the inflammatory biomarkers cfDNA, HMGB1, and PCT are increased in canine GDV. Only lactate and PCT concentrations were prognostic in this population of GDV dogs and were predictive of the presence of gastric necrosis and of non-survival to hospital discharge, respectively