The Cardiac Biomarker NT-proBNP Is Increased in Dogs with Azotemia

Background: Amino‐terminal probrain natriuretic peptide (NT‐proBNP) has been proposed as a useful biomarker for heart disease in dogs. In humans, decreased glomerular filtration rate (GFR) increases NT‐proBNP. Objective: To investigate whether decreased GFR as indicated by plasma creatinine concentration is associated with increased NT‐proBNP in dogs without heart disease. Animals: Four groups of dogs: healthy (n= 39), azotemic (n= 36), heart disease (n= 37), and congestive heart failure (CHF) (n= 7) presented to 2 teaching hospitals. Methods: Prospective observational cohort study. Plasma creatinine concentration and NT‐proBNP were measured in every dog. Nonparametric tests were used to compare the differences among groups. The median and actual results for each group were compared with the manufacturer's recommended and previously published suggestions for cut‐off values for diagnosis of heart disease. Results: Median (range) plasma creatinine concentration was 1.47 (1.06–1.70), 4.36 (1.74–15.6), 1.22 (0.69–1.91), and 1.45 (0.63–1.64) mg/dL and median (range) NT‐proBNP was 118 (2–673), 556 (37–1,819), 929 (212–5,658), and 3,144 (432–5,500) pmol/L for the healthy, azotemic, heart disease, and CHF groups, respectively. Pair‐wise comparison indicated a significant difference among all groups for NT‐proBNP (P≤ .049). Plasma creatinine concentration was significantly higher in the azotemic group compared with other groups (P < .001) but there was no significant among other groups. Application of 3 recommended cut‐off values led to misclassification of dogs with azotemia as having heart disease. Conclusions: Azotemia results in NT‐proBNP being increased to concentrations reported as diagnostic of heart disease or heart failure in dogs. Care should be employed when interpreting the results of NT‐proBNP in patients with known or possible increased plasma creatinine concentration.     

Cloning and tissue expression of the equine transferrin receptor

Background: Characterization of anemia in horses presents a challenge, as they do not release reticulocytes into peripheral blood. Transferrin receptor (TfR) expression is highest on erythroid cells in people and rats, and measurement of a soluble serum form (sTfR) is used to quantify erythropoiesis in these species. We hypothesized that equine TfR (eTfR) expression is similar in quantity and distribution to that in these other species and thus has potential for characterization of the regenerative response in anemic horses. Objective: This study was conducted to clone and sequence the eTfR gene and measure expression levels using quantitative real‐time PCR and immunohistochemical (IHC) staining. Methods: Total RNA from equine bone marrow was used to produce cDNA. The eTfR gene was amplified using pooled gene‐specific primers, and PCR products were sequenced. Rapid amplification of cDNA ends was used to obtain the first 22 nucleotides of the coding sequence. Quantitative PCR was performed using eTfR gene‐specific primers, and IHC staining was used to localize TfR protein expression. Results: The deduced amino acid (aa) sequence (767 aa) of the eTfR was 75–83% identical with sequences of the receptor in several other mammals. As in people and rats, eTfR mRNA expression was highest in the bone marrow, and distribution in other tissues was also similar. Conclusion: The eTfR gene is similar to that of other mammals in structure and expression levels. We hypothesize that it is also similar in function and that, following development of an immunoassay, determining sTfR concentrations will be useful for identifying the regenerative response in anemic horses.     

In vitro Inhibition of Canine Complement‐Mediated Hemolysis

Background

Immune‐mediated hemolytic anemia (IMHA) is the most common hematologic immune‐mediated disease in dogs. Complement fixation on erythrocytes causes hemolysis. Complement inhibition decreases hemolysis in people with the hemolytic disease and also may prove effective in treating IMHA in dogs.

Hypothesis/Objectives

Evaluate the in vitro efficacy of 2 complement inhibitors used in humans against canine complement.

Methods

The inhibitory activity of the C3‐inhibitor compstatin and recombinant human C1‐esterase inhibitor (C1‐INH) was evaluated using an in vitro hemolytic assay and spectrophotometric measurement of released hemoglobin. Dose‐response curves for each inhibitor were generated.

Results

Compstatin decreased approximately 50% of canine complement‐mediated hemolysis in initial experiments. This inhibition largely was lost when a new lot of drug was purchased. C1‐INH showed a dose‐dependent inhibition. The highest concentration of C1‐INH tested (500 μg/mL) decreased >80% of canine complement‐mediated hemolysis, and the lowest concentration tested (31.25 μg/mL) decreased hemolysis >60%.

Conclusions and Clinical Importance

Human C1‐INH is a robust inhibitor of canine complement‐mediated hemolysis, whereas compstatin was minimally and variably effective. Human C1‐INH may substantially decrease complement‐mediated hemolysis in dogs with IMHA and warrants further investigation.     

Study on efficacy of the decoction of invigorating kidney and activating blood circulation in BALB/C mice with immune mediated aplastic anemia

AIM and METHODS: To study the efficacy of the decoction of invigorating kidney and activating blood circulation(DIKABC) in BALB/C mice with immune mediated aplastic anemia. 50 female BALB/C mice at 8~12 weeks of age were divided into 5 groups:(1) control,(2) model,(3) model+DIKABC(H), (4) model+DIKABC(L), and (5) model+ciclosporin A(CsA). The model of immune mediated aplastic anemia in BALB/C mice, which received sublethal whole body irradiation, lymph node and thymus cells of DBA/2 mice, was given different liquid from groups and administered appropriately for 9 days, blood cells and bone marrow nucleated cells(BMNC) and CD₄₅⁺ cells were determined by flow cytomytry(FCM).RESULTS: RBC,WBC,Platelet,BMNC in DIKABC groups were higher than those in model group (P＜0.01/0.05), but those in CsA group were lower than those in DIKABC groups (P＜0.05).Compared with model group, CD₄₅⁺ cells in DIKABC groups increased (P＜0.05),but were lower than those in control groups (P＜0.05). CONCLUSION: The decoction of invigorating kidney and activating blood circulation could increase the percentages of CD₄₅⁺ cells in bone marrow and prevent immune mediated aplastic anemia in BALB/C mice.