Urinary Neutrophil Gelatinase-associated Lipocalin as a Marker for Identification of Acute Kidney Injury and Recovery in Dogs with Gentamicin-induced Nephrotoxicity.

BACKGROUND: Acute kidney injury (AKI) is associated with high mortality rates in dogs, which may be a consequence of late recognition using traditional diagnostic tests. Neutrophil gelatinase-associated lipocalin (NGAL) is a protein-induced during kidney injury that may identify AKI earlier than traditional tests. OBJECTIVES/HYPOTHESIS: To evaluate urinary NGAL (uNGAL) and uNGAL-to-urinary creatinine ratio (UNCR) as early markers of kidney injury and recovery in an AKI model in dogs. It was hypothesized that these markers would document AKI earlier than serum creatinine concentration. ANIMALS: Five purpose-bred dogs. METHODS: Prospective study. Acute kidney injury, defined as a > 50% increase in serum creatinine concentration above baseline, was induced in dogs by gentamicin administration (8-10 mg/kg SC q8h). Blood and urine collected for biochemical analyses and uNGAL and urinary creatinine concentrations, respectively, during AKI induction and recovery. RESULTS: Acute kidney injury was diagnosed significantly earlier based on a 7-fold increase in UNCR compared to a > 50% increase in serum creatinine concentration (day 8; range, 2-10 mg/dl vs day 16; range, 14-19 mg/dl; P = .009). During recovery, the initial decrease in UNCR preceded the decrease in serum creatinine concentration by a median of 2 days. The uNGAL changes paralleled UNCR changes, but the increase in uNGAL was triphasic; the initial peak occurred earlier than UNCR (median, day 11 versus median, day 19). CONCLUSIONS AND CLINICAL IMPORTANCE: The UNCR was early marker of gentamicin-induced AKI and its decrease documented onset of renal recovery. Additional studies are needed to validate this marker in dogs with naturally occurring renal injury.

5-Lipoxygenase expression in benign and malignant canine prostate tissues.

5-Lipoxygenase (5-LO) is overexpressed in human prostate carcinomas (PCs), and its inhibition decreases proliferation and induces apoptosis in prostate cancer cell lines. We hypothesized that 5-LO would be overexpressed in canine PC compared with benign prostate tissue and may be important in the pathogenesis of the disease. Immunoblot analysis of canine PC and benign prostatic hyperplasia (BPH) tissues demonstrated 5-LO expression in both. 5-LO immunohistochemical staining was not significantly different within the stromal or epithelial components of canine primary PC, BPH or suppurative prostatitis, suggesting that differential expression of this enzyme does not occur in these conditions. The percentage of tumour cells expressing 5-LO was significantly lower in metastatic PC lesions compared with primary PC (P < 0.0001). This decreased expression may indicate down-regulation or altered expression of the enzyme with progression of canine PC to a metastatic phenotype.

Evaluation of the positive predictive value of serum protein electrophoresis beta-gamma bridging for hepatic disease in three domestic animal species.

Beta-gamma bridging (ß-γ bridging) on serum protein electrophoresis is touted as being virtually pathognomonic for hepatic disease. However, the criteria for ß-γ bridging are not defined, and few publications support a relationship between ß-γ bridging and liver disease. The goal of this retrospective study was to evaluate the prevalence of hepatic pathology in animals with ß-γ bridging. All serum protein electrophoretograms from clinical patients generated at the University of Georgia between 1994 and 2008 were evaluated for the presence of ß-γ bridging, defined as (1) an albumin:globulin ratio below the reference interval; (2) indistinct separation between all ß and γ globulin fractions or between the ß(2) and γ fractions, with a negative shoulder slope of < 5%; and (3) predominance of γ proteins versus ß proteins. Of the 237 electrophoretograms examined, 25 (11 dogs, 11 cats, 3 horses) met the inclusion criteria for ß-γ bridging. Patients were classified into disease categories on the basis of biochemical, cytologic, and/or histologic findings. Positive predictive values of ß-γ bridging for hepatic and infectious diseases were determined with a one-sided exact binomial test. Of 25 animals, 8 had evidence for hepatic disease, whereas 9 had infectious diseases. As such, the positive predictive value of ß-γ bridging for hepatic disease was 32.0%, with a 95% confidence interval of 15.0% to 53.5% (P < .001), whereas for infectious disease, the positive predictive value was 36.0%, with a similar confidence interval. Beta-gamma bridging is not pathognomonic for liver diseases and is as frequently found with infectious diseases.

Mechlorethamine, procarbazine and prednisone for the treatment of resistant lymphoma in dogs.

Forty-one dogs with resistant lymphoma were treated with a modified MOPP (mechlorethamine, vincristine, procarbazine and prednisone) protocol (MPP [mechlorethamine, procarbazine and prednisone] administered on a 21-day cycle, shortened from the 28-day MOPP cycle). The overall response rate to MPP was 34% for a median of 56 days (95% confidence interval 30-238). Seventeen percent of dogs had a complete response for a median duration of 238 days, 17% had a partial response for a median of 56 days and 32% had stable disease for a median of 24 days. Histological grade or cell morphology on cytology was associated with response. Minimal toxicity was observed with the MPP protocol, suggesting that further dose intensification or addition of another chemotherapeutic agent would be possible.

Effects of 1,25(OH)2D3, 25OHD3, and EB1089 on cell growth and Vitamin D receptor mRNA and 1alpha-hydroxylase mRNA expression in primary cultures of the canine prostate.

The aim of this study was to investigate effects of 1,25(OH)(2)D(3) (calcitriol), 25OHD(3), and EB1089 on cell growth and on Vitamin D receptor (VDR) mRNA and 1alpha-hydroxylase (1alpha-OHase) mRNA expression in normal canine prostatic primary cultures. Canine prostatic epithelial cells were isolated, cultured, and treated with vehicle (ethanol), calcitriol, 25OHD(3), and EB1089 at 10(-9) and 10(-7)M. The VDR was present in epithelial and stromal cells of the canine prostate gland. 1,25(OH)(2)D(3), 25OHD(3), and EB1089 inhibited epithelial cell growth at 10(-7)M compared to vehicle-treated controls [calcitriol (P < 0.01), EB1089 (P < 0.01), and 25OHD(3) (P < 0.05)]. Epithelial cells treated with calcitriol and EB1089 at 10(-7)M had slightly increased VDR mRNA expression (0.2-0.3-fold) at 6 and 12h compared to controls. There was no difference in 1alpha-OHase mRNA expression in epithelial cells treated with these three compounds. 1,25(OH)(2)D(3) and its analogs may be effective antiproliferative agents of epithelial cells in certain types of prostate cancer.

Canine prostate carcinomas express markers of urothelial and prostatic differentiation.

Prostate carcinoma and transitional cell carcinoma (TCC) occur in the prostate gland of older dogs and have morphologic similarities when evaluated by light microscopy. The dog is a commonly used animal model for studying human prostate carcinoma; therefore, it is important to accurately differentiate canine prostate carcinomas from TCCs. We investigated whether keratin 7 (K7) and arginine esterase (AE) would aid differentiation of canine prostate carcinoma from TCC. K7 expression was evaluated in normal and neoplastic canine prostate and bladder tissues using immunohistochemistry. The expression of AE messenger ribonucleic acid (mRNA) in normal and neoplastic canine prostate and bladder was detected using northern blots and reverse transcriptase-polymerase chain reaction (RT-PCR). In addition, AE enzyme activity was measured in normal and neoplastic canine prostate and bladder tissues. We found marked similarities in K7 expression in prostate carcinomas and TCCs. AE mRNA was present in high levels in normal prostatic tissue but was reduced in prostate carcinoma by northern blot assay. Nested RT-PCR detected AE mRNA both in TCCs (13 of 15) and in prostate carcinomas (13 of 13). Enzymatic activity of AE was high in normal prostate gland and in some prostate carcinomas, whereas normal bladder and TCCs produced lower levels of AE. In conclusion, K7 and AE cannot be used to differentiate TCC from prostate carcinoma in dogs.

Results of intradermal tests in horses without atopy and horses with atopic dermatitis or recurrent urticaria.

OBJECTIVE: To compare results of intradermal tests (IDT) for environmental allergens at 30 minutes and 4, 6, and 24 hours after injection in horses without atopy and horses with atopic dermatitis (AD) or recurrent urticaria (RU). ANIMALS: 22 horses without atopy, 10 horses with RU, and 7 horses with AD. PROCEDURE: In all horses, medical history was obtained, and results of physical examination, hematologic examination, serum biochemical analyses, examination of bronchoalveolar lavage fluid, and IDT with 73 allergens were examined. RESULTS: Horses with AD or RU had a significantly greater mean number of positive reactions for IDT, compared with horses without atopy. Horses with AD had a significantly greater number of positive reactions than horses without atopy in every allergen group at all time periods, except for molds at 4 and 24 hours. Horses with RU had a significantly greater number of positive reactions than horses without atopy in every allergen group, except for molds at 30 minutes and 4 and 6 hours, trees at 4 and 6 hours, and grasses at 4 hours. CONCLUSIONS AND CLINICAL RELEVANCE: A significantly greater number of positive reactions for IDT in horses with AD or RU, compared with horses without atopy, provides evidence of type-I IgE-mediated hypersensitivity for these diseases. Evaluation of results of IDT performed in horses with AD or RU is useful in determining specific allergens for the formulation of immunotherapy along with providing identification of allergens that could be useful when creating avoidance strategies.

Comparison of immediate intradermal test reactivity with serum IgE quantitation by use of a radioallergosorbent test and two ELISA in horses with and without atopy.

OBJECTIVE: To compare a radioallergosorbent test and 2 ELISA with intradermal testing for the determination of environmental allergen hypersensitivity in horses with and without atopic diseases. DESIGN: Prospective clinical study. ANIMALS: 10 horses with recurrent urticaria, 7 with atopic dermatitis, 16 with chronic obstructive pulmonary disease, and 22 without atopy. PROCEDURE: History, physical examination, hemogram, serum biochemical analyses, bronchoalveolar lavage, and an intradermal test (used as the criterion standard) with a regional panel of 73 allergens were performed in all horses. Serum was analyzed by use of the 3 in vitro assays of allergen-specific IgE. RESULTS: An ELISA based on the alpha chain of the high-affinity IgE receptor, the Fcepsilon receptor immunoglobin epsilon chain (FcepsilonRIalpha) for IgE, had the overall highest kappa statistic (0.238), positive predictive value (49%), and negative predictive value (78%). Overall agreement between the FcepsilonRIalpha-based ELISA and the intradermal test was fair. The highest kappa statistic was obtained by the FcepsilonRIalpha-based ELISA in horses with atopic dermatitis (0.330). Kappa statistics for the radioallergosorbent test and a polyclonal antibody-based ELISA agreed slightly with that of the intradermal test at best. CONCLUSIONS AND CLINICAL RELEVANCE: None of the 3 serum allergy tests reliably detected allergen hypersensitivity, compared with the intradermal test. The FcepsilonRIalpha-based ELISA performed significantly better overall than the other 2 tests. Low sensitivity of all 3 assays indicates the need for continued study to elucidate a more sensitive test for the determination of potentially pathogenic allergens in horses.