A comprehensive analysis of albuminuria in canine chronic kidney disease.

BACKGROUND: Albuminuria, an important marker of decreased kidney function in chronic kidney disease (CKD), is not routinely used for CKD detection or proteinuria appearance. Its relationships with biochemical parameters and blood pressure in dogs are poorly understood. OBJECTIVES: This study aimed to evaluate the relationship of albuminuria with various CKD markers, its correlation with the urinary protein to creatinine ratio (UPC), and hypertension in dogs with early stages of CKD. It also sought to determine the usability of the urinary albumin to creatinine ratio (UAC) for CKD screening. METHODS: The study reviewed records of 102 dogs, categorising them into four groups based on disease status. UAC and UPC ratio, biochemistry and haematology variables, age, and systolic blood pressure were determined. RESULTS: The Pearson's correlation coefficient between log-transformed values of UPC and UAC was r = 0.902 (95% CI: 0.87 to 0.93). Median UAC ratio values were 2.1 mg/g for the Healthy control group (n = 17), 54.2 mg/g for early stages CKD (n = 42), 5.8 mg/g for Acute sick control (n = 30), and 104 mg/g for Chronic sick control (n = 13). Thresholding UAC ratio as an indicator for impaired kidney function with the threshold of 10 mg/g (established based on the receiver operating characteristic curve) had a sensitivity 81.8%, specificity of 89.4%, positive predictive value (PPV) 90%, and negative predictive value (NPV) 80.1%. The correlation of UAC with biochemistry and haematology variables was statistically significant; for SDMA (µg/L), it was r = 0.566 and for other variables, it was weak to moderate. UAC was markedly elevated in cases of severe hypertension. CONCLUSIONS: UAC ratio was significantly different among dogs with impaired and not impaired kidney function. The correlation strength for the UAC and UPC ratios was high. UAC ratio may be a promising marker for proteinuria analysis in dogs with CKD or other kidney function alterations.

Regulatory T lymphocytes and selected cytokines in horses with mild to severe equine asthma and different inflammatory cytology profiles.

This study determined Tregs and inflammatory cytokines in BALF and peripheral blood (PB) of adult horses with mild and severe asthma and different BALF cytological inflammation profiles. Horses of diverse breeds with asthma (age range: 2-20 years, n = 24) were divided into groups according to the number of points obtained in a standardized clinical scoring system (mild-moderate equine asthma - MEA, severe equine asthma - SEA) and according to the inflammation type based on cytological finding. Plasma levels of IFN-γ, IL-4, IL-10, IL-17 and MMP-9 in the BALF were determined by ELISA. Tregs in the BALF and PB were detected by flow cytometry. Horses with SEA were older than horses with MEA and had higher concentrations of MMP-9 in the BALF (P < 0.05). The neutrophilic inflammation group was characterised by higher age, grade of clinical score, percentage of Tregs and MMP-9 concentration in the BALF compared to the mastocytic inflammation group (P < 0.05). Age positively correlated with neutrophils, grade of score, Tregs in the BALF and in the PB. The grade of score positively correlated with MMP-9. The percentage of Tregs in the BALF positively correlated with Tregs in the PB. Tregs in the BALF and PB positively correlated with neutrophils and MMP-9 in the BALF and with IFN-γ, IL-4 and IL-10 in the plasma. IFN-γ and IL-10 in the plasma positively correlated with the age. The results indicate that the evaluation of selected parameters in PB, the collection of which is less invasive than the collection of BALF, is of potential importance.

Lymphocyte immunophenotyping and concentration of MMP-9 in transudates and exudates in horses.

This study is the first to provide information on the lymphocyte subpopulations in peritoneal effusions in horses. Peritoneal transudates (n = 12), peritoneal exudates (n = 6) and a pleural exudate (n = 1) were analyzed. The total nucleated cell count (TNCC), total protein (TP) and matrix metalloproteinase-9 (MMP-9) concentration determined by ELISA were measured and routine cytological evaluation was performed. CD3, CD4, CD8 and CD21 positive cells were detected by flow cytometry. A higher percentage of neutrophils (P < 0.05) and higher MMP-9 (P < 0.01) levels were found in exudates. A higher percentage of macrophages (P < 0.05) and lymphocytes (P < 0.01) were found in transudates. CD4 + lymphocytes were the most common lymphocyte subpopulation in all samples. CD21 + lymphocytes were the least common in all samples. A large variability in the percentage of CD21 + lymphocytes was found in exudates. The percentage of CD21 + lymphocytes positively correlated with the level of total protein (r = 0.5704, P < 0.05). The correlation was even stronger in the group of exudates. The percentages of lymphocyte subpopulations did not correlate with the level of MMP-9 or with cytological findings. The level of MMP-9 positively correlated with the percentage of neutrophils (r = 0.4980, P < 0.05), the level of TP (r = 0.7855, P < 0.01) and TNCC (r = 0.6129, P < 0.01). A significantly higher level of MMP-9 was detected in euthanized horses than in horses that survived (P < 0.05). However, it was shown that the level of MMP-9 in the peritoneal fluid can change significantly in a short time. More studies on repeated abdominocentesis could contribute to elucidating the role of MMP-9 as a prognostic indicator.

Lymphocyte immunophenotyping in dogs with lymphopenia of common causes.

Lymphocyte immunophenotyping can be useful for evaluating immune competence and predicting the disease prognosis. It is essential to gain knowledge about canine lymphocyte immunophenotypes in various conditions. The study deals with the characteristics of lymphopenia in dogs, with an emphasis on lymphocyte immunophenotyping by flow cytometry. Blood samples from 44 dogs with lymphopenia were included in the study. All lymphopenias sent from veterinary clinics to the diagnostic laboratory were analyzed. The hematological and biochemical abnormalities were investigated, as well as the effect of the age. Lymphopenias were classified according to the level of C-reactive protein (CRP). The percentage of T cells, B cells, Th cells and Tc cells, and T/B and Th/Tc ratios were determined by flow cytometry. Lymphopenias often occurred in dogs over 7 years of age (79.5 %). The most common were postoperative lymphopenia (31.8 %) and inflammatory diseases (29.5 %), most commonly affecting the gastrointestinal tract. Frequent abnormalities were monocytosis (56.8 %), increased CRP (72.7 %) and decreased albumin/globulin ratio (50.0 %). The percentage of Th lymphocytes was significantly lower in the group with elevated CRP than in the group with basal CRP (P = 0.0329). A negative correlation was found between the level of CRP and the percentage of Th lymphocytes (r = -0.3278, P = 0.0390). This study provided new insights into the appearance, incidence and classification of canine lymphopenia.

Changes in rabbit lymphocyte subpopulations and activation following long-term administration of meloxicam.

Meloxicam is a commonly used analgesic in rabbits. However, its possible impact on lymphocyte subpopulations remained unknown. The aim of the study was to investigate a possible effect of long-term administration of meloxicam on rabbit lymphocyte subpopulations. The study included 8 rabbits given meloxicam orally once daily (1 mg/kg BW) for 14 days and 8 rabbits as a control group. Peripheral blood samples were collected on day 0 (before the first dose of meloxicam), day 3, 7 and 14. Samples were evaluated with a haematology analyser and a flow cytometer. A significant decrease in T: B cell ratio was found in all samples taken during meloxicam administration compared to day 0, as well as in comparison with the control group (P < 0.01). A significant increase (P < 0.05) in proportion of CD5 +CD8 + lymphocytes occurred by day 3. Subsequently, although the values slightly decreased, they still remained elevated throughout all the experiment compared to the values from day 0 (P < 0.05). A slight decrease in T and B cell activation (CD5 +CD25 + and IgM+CD25 +) noticed by day 3, declined during the next days of administration and became more and more significant (finally, P = 0.0078). Since a high significant decrease (P < 0.01) in both T and B cell activation as well as a significant increase (P < 0.05) in CD5 +CD8 + T cells proportion were observed after meloxicam administration, a predicted effect of long-term administration of meloxicam on rabbit lymphocytes was confirmed.

Electrolyte, Oestradiol and Progesterone Concentrations in Ovarian Cyst Fluid in Guinea Pigs (Cavia porcellus).

Despite their high prevalence, the aetiopathogenesis of ovarian cysts in guinea pigs (Cavia porcellus) is unknown. We examined tissue samples of cystic ovaries from 20 guinea pigs by histopathology and immunohistochemistry for CD10 antigen, and determined the concentrations of Na+, K+, Cl-, oestradiol and progesterone in the cyst fluid. Cystic rete ovarii were found in all 20 guinea pigs but no follicular cysts. Concentrations of Na+ (142.2 ± 5.04 mmol/l), K+ (5.6 ± 0.76 mmol/l) and Cl- (116.1 ± 5.25 mmol/l) corresponded to extracellular, rather than intracellular, fluid indicating that rete ovarii cysts in guinea pigs are transudative in nature. A positive correlation was found between Na+ and Cl- concentrations (r = 0.7713, P <0.0001). The mean concentrations of oestradiol (61.9 ± 17.44 pg/ml) and progesterone (2.7 ± 5.42 ng/ml) in the cyst fluid were higher than expected because the rete epithelium is not known to have steroidogenic activity. A negative correlation was found between oestradiol and progesterone concentrations in cyst fluid (r = -0.4296, P = 0.0178). Possible pathogenetic mechanisms of cystic rete ovarii in guinea pigs are discussed.

Acute Pneumonia Caused by Clinically Isolated Legionella pneumophila Sg 1, ST 62: Host Responses and Pathologies in Mice.

Legionnaires' disease is a severe form of lung infection caused by bacteria belonging to the genus Legionella. The disease severity depends on both host immunity and L. pneumophila virulence. The objective of this study was to describe the pathological spectrum of acute pneumonia caused by a virulent clinical isolate of L. pneumophila serogroup 1, sequence type 62. In A/JOlaHsd mice, we compared two infectious doses, namely, 104 and 106 CFU, and their impact on the mouse status, bacterial clearance, lung pathology, and blood count parameters was studied. Acute pneumonia resembling Legionnaires' disease has been described in detail.

Effects of the Administration of Different Buffered Balanced Crystalloid Solutions on Acid-Base and Electrolyte Status in Dogs with Gastric Dilation-Volvulus Syndrome: A Randomized Clinical Trial.

OBJECTIVE: To investigate the effect of three different buffered balanced crystalloid solutions on acid-base status and electrolyte concentrations in dogs with gastric dilation-volvulus (GDV) syndrome. METHODS: The study design was a prospective, randomized clinical trial of 40 dogs. The dogs were randomly assigned to one of three groups according to the fluid used: Hartmann's solution (H), Plasmalyte (PL), and Ringerfundin (RF). Hemoglobin, albumin, lactate, electrolyte, and acid-base parameters were determined before fluid administration (T0) and at the end of surgery (T1). Results were assessed by one-way ANOVA, Fisher's exact test, the Wilcoxon signed-rank test, the Kruskal-Wallis test, and a linear mixed-effect regression model. A significance level of 0.05 was used in all analyses. RESULTS: Bicarbonate and base excess (BE) levels increased and chloride concentration decreased in the PL group; in contrast, strong ion difference apparent (SIDapp) decreased and chloride concentration increased in the RF group. The mixed-effect model confirmed a significant interaction between the type of solution and time on the changes in bicarbonate, BE, anion gap (AG), SIDapp, and chloride levels. CLINICAL SIGNIFICANCE: Significantly different effects in acid-base parameters were observed in dogs after intravenous administration of H, PL, and RF. However, clinical significance of these changes is lacking, requiring further investigation in a larger randomized controlled clinical trial.

Acid-Base, Electrolyte and Lactate Abnormalities as Well as Gastric Necrosis and Survival in Dogs With Gastric Dilation-Volvulus Syndrome. A Retrospective Study in 75 Dogs.

Gastric dilation-volvulus (GDV) syndrome in dogs is associated with complex metabolic, acid-base, and electrolyte abnormalities. The aim of this study was to evaluate previously analyzed factors (lactate and BE) in combination with other acid-base parameters (pH, pCO2, bicarbonate, base excess [BE], anion gap [AG], and strong ion difference) and electrolyte concentrations and to evaluate their association with the incidence of gastric necrosis and outcome in dogs with GDV. A retrospective study in 75 dogs with gastric dilation-volvulus syndrome, University veterinary teaching hospital. Medical records were reviewed including signalment, history, initial plasma lactate, acid-base parameters, and electrolyte concentrations, surgical findings and outcome. The overall mortality was 18.7%. In dogs with gastric necrosis, higher initial plasma lactate (median 5.84 vs. 3.36 mmol/L) and AG (20.7 vs. 16.55 mmol/L) and lower pH (7.29 vs. 7.36), bicarbonate (18.7 vs. 22.9 mmol/L), and BE concentration (-8.1 vs. -1.85 mmol/L) were found compared to dogs without gastric necrosis. Anorganic phosphorus was the only electrolyte investigated for which a significant difference was noted between dogs with and without gastric necrosis (1.93 vs. 1.39 mmol/L). The initial plasma lactate concentration (3.36 mmol/L vs. 9.68 mmol/L) and AG (16.8 vs. 20.95 mmol/L) were lower in survivors than nonsurvivors. Survivors had higher pH (7.35 vs. 7.27), bicarbonate concentrations (22.9 vs. 17.35 mmol/L), and BE (-1.9 vs. -9.55 mmol/L) compared to nonsurvivors. Anorganic phosphorus was ultimately the only electrolyte with a significant difference between survivors and nonsurvivors (1.4 vs. 1.84 mmol/L). A multivariate logistic regression model of combination lactate, pH, bicarbonate, BE, AG, and anorganic phosphorus identified pH ≤7.331 and bicarbonate as factors independently associated with gastric necrosis. Similarly, pH ˃7.331, bicarbonate and anorganic phosphorus were independently associated with outcome. Higher initial plasma lactate, AG and anorganic phosphorus levels, and lower pH, BE and bicarbonate concentrations were found in GDV dogs with gastric necrosis. Similarly, initially higher plasma lactate, AG and anorganic phosphorus concentrations, and lower pH, BE and bicarbonate were found in GDV dogs who required euthanasia or who died after surgery. Of these parameters, pH and bicarbonate were strongly and independently associated with gastric necrosis, and pH, bicarbonate and phosphorus were independently associated with outcome.

Determination of reference intervals of acid-base parameters in clinically healthy dogs.

OBJECTIVE: To establish reference intervals for traditionally- and Stewart's approach-determined acid-base parameters in a population of clinically healthy dogs. DESIGN: Prospective study (June 2011-September 2012). SETTING: Veterinary teaching hospital. ANIMALS: Two hundred twenty-four client-owned, clinically healthy dogs. INTERVENTIONS: Blood was collected from the jugular vein and the dorsal pedal artery. MEASUREMENTS AND MAIN RESULTS: In the whole blood samples, pH, PCO2 , and PO2 were measured and HCO3- , standard and total bicarbonate, base excess, oxygen content, and alveolar-arterial oxygen differences were calculated. Plasma sodium, potassium, chloride, calcium, albumin, and lactate concentrations were measured and the following parameters were calculated separately for venous and arterial samples: anion gap, anion gap corrected for albumin and phosphate, sodium chloride difference and ratio, chloride gap, strong ion difference, strong ion gap, and unmeasured anions. CONCLUSION: Reference intervals for traditionally- and Stewart's approach-determined acid-base parameters were established for venous and arterial blood in dogs.

C-reactive protein and high mobility group box 1 in dogs with gastric dilatation and volvulus.

OBJECTIVE: To (1) measure C-reactive protein (CRP) and high mobility group box 1 (HMGB1) and (2) evaluate their prognostic value and relationship to severity of systemic inflammatory response syndrome, routine hematological and acid-base parameters in dogs with gastric dilatation volvulus (GDV). DESIGN: Prospective observational study from September 2010 to June 2012. SETTING: Veterinary teaching hospital. ANIMALS: Forty-one client-owned dogs with GDV. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Blood was collected before surgery (baseline), postsurgery, 6-10 hours postsurgery, and 18-22 hours postsurgery. CRP and HMGB1 were measured in all samples, and routine hematological, biochemical, and acid-base analyses were performed. Only baseline and postsurgery samples were used from nonsurvivors (n = 10). CRP increased significantly from postsurgery sampling to 18-22 hours postsurgery, while HMGB1 did not change over time. There was a significant difference in HMGB1 between survivors and nonsurvivors over time. Both proteins correlated with systemic inflammatory response syndrome severity, total leukocyte, segmented neutrophils, and band counts. HMGB1 correlated also with acid-base parameters (pH, bicarbonate, base excess). CONCLUSION: HMGB1 and CRP behaved differently in regards to their kinetic patterns, with HMGB1 appearing to better reflect the severity of tissue injury in dogs with GDV than CRP.

Retention of nanoparticles-labeled bone marrow mononuclear cells in the isolated ex vivo perfused heart after myocardial infarction in animal model.

Cell therapy of myocardial infarction (MI) is under clinical investigation, yet little is known about its underlying mechanism of function. Our aims were to induce a sub-lethal myocardial infarction in a rabbit, to evaluate the abilities of labeled bone marrow mononuclear cells to migrate from the vessel bed into extracellular space of the myocardium, and to evaluate the short-term distribution of cells in the damaged left ventricle. Sub-lethal myocardial infarction was induced in rabbits by ligation of the left coronary vessel branch (in vivo). The Langendorff heart perfusion model (ex vivo) was used in the next phase. The hearts subjected to MI induction were divided into 3 groups (P1-P3), and hearts without MI formed a control group (C). Nanoparticles-labeled bone marrow mononuclear cells were injected into coronary arteries via the aorta. Perfusion after application lasted 2 minutes in the P1 group, 10 minutes in the P2 and C groups, and 25 minutes in the P3 group. The myocardium of the left ventricle was examined histologically, and the numbers of labeled cells in vessels, myocardium, and combined were determined. The numbers of detected cells in the P1 and C groups were significantly lower than in the P2 and P3 groups. In the P2 and P3 groups, the numbers of cells found distally from the ligation were significantly higher than proximally from the ligation site. Bone marrow mononuclear cells labeled with iron oxide nanoparticles proved the ability to migrate in the myocardium interstitium with significantly higher affinity for the tissue damaged by infarction.