Vinblastine/prednisolone chemotherapy leads to hematological toxicity in dogs with high-grade or metastatic mast cell tumors.

OBJECTIVE: To determine the myelosuppressive effects/hematological toxicities, other general toxicities, and when these occur during vinblastine/prednisolone chemotherapy in dogs bearing high-grade or metastatic cutaneous/subcutaneous mast cell tumors (MCTs). METHODS: Medical records were retrospectively reviewed between November 1, 2016, and March 1, 2023. Thirty client-owned dogs with histopathologically confirmed cutaneous high-grade MCTs/metastatic subcutaneous MCTs and that subsequently completed a 12-week vinblastine/prednisolone chemotherapy protocol were included. Hematology was assessed before commencing chemotherapy and before each vinblastine treatment. The effect of each treatment upon hematological values was evaluated. Measured outcomes included the type, frequency, and severity of hematological and other more general toxicities. RESULTS: 24 of 30 dogs experienced at least 1 hematological toxicity, 6 experienced gastrointestinal toxicity, and 4 experienced lethargy. The most common toxicity was anemia (15/30 [50%]), with 93.3% (14/15 dogs) classified as Veterinary Cooperative Oncology Group-Common Terminology Criteria for Adverse Events grade I and 6.6% (1/15) classified as grade II. The second most common toxicity was neutropenia (14/30 [46.6%]), with 71.4% (10/14) classified as grade I and 28.6% (4/14) as grade III. The least common hematological toxicity was thrombocytopenia (4/30 [13%]), all grade I. Neutropenia mainly occurred during weeks 2 and 3; however, there was no significant decrease in neutrophil count relative to baseline. Neutrophil count increased and Hct decreased during weeks 6 to 12 of treatment when compared to baseline. No change in platelet count was observed. CLINICAL RELEVANCE: Vinblastine/prednisolone chemotherapy leads to hematological toxicity; however, this was mostly low-grade and did not require major intervention. Vinblastine/prednisolone chemotherapy is well tolerated in dogs bearing high-grade or metastatic MCTs.

Serum C-reactive protein concentrations in dogs with structural and idiopathic epilepsy.

BACKGROUND: C-reactive protein (CRP) is an acute-phase protein produced by the liver during systemic inflammation. In humans, some epilepsies are associated with increased serum CRP (sCRP) concentrations, but this has yet to be proven in veterinary studies. Dogs with structural epilepsy (SE) and normal interictal neurological examination are hard to distinguish from dogs with idiopathic epilepsy (IE) without the use of advanced imaging. METHODS: The study included eight dogs with SE and 12 dogs with IE from a referral hospital population. This was a retrospective observational cohort study. The Mann-Whitney test was used to compare the sCRP concentrations within 24 hours of the last epileptic seizure between dogs with SE or IE. RESULTS: Dogs with SE had higher sCRP concentrations than dogs with IE (8.9 [range <2.2-53.2] mg/L vs. <2.2 [range <2.2-6.9] mg/L; p = 0.043). Five of the eight (62%) dogs with SE had an sCRP concentration above the reference interval, compared with none of the 12 dogs with IE. LIMITATIONS: The small sample size was the major limitation of this study. Other inflammatory causes were also not exclusively ruled out, although further clinical investigations were not indicated. CONCLUSIONS: This study found that sCRP concentrations were higher in this cohort of dogs with SE than in those with IE. Further studies with larger cohorts of dogs are warranted to validate if sCRP can be used as an additional biomarker for SE.

Prevalence of neoplasia and concurrent diseases in dogs and cats with hypercobalaminemia: A retrospective case-control study.

BACKGROUND: Hypercobalaminemia is infrequently reported in companion animals and is considered of low clinical significance. Recent studies have described its association with inflammatory, immune-mediated, endocrine, and neoplastic conditions in dogs and cats. OBJECTIVES: We aimed to investigate the association between hypercobalaminemia and neoplasia in companion animals and to identify other concurrent diseases or clinicopathologic changes. METHODS: This is a retrospective, case-control study. Medical records of patients with measured serum cobalamin concentration (2015-2020) and no history of prior supplementation were reviewed. Hypocobalaminemic animals were excluded. Variables were compared between groups (hypercobalaminemic vs. normocobalaminemic) using non-parametric statistics. Data are presented as median (range). RESULTS: Thirty-five dogs and eight cats were hypercobalaminemic. At baseline, neoplasia was confirmed in 4/35 hypercobalaminemic dogs versus 11/70 control dogs (P = 0.77) and 0/8 hypercobalaminemic cats versus 3/16 control cats (P = 0.53). Cases without neoplasia at baseline were followed for 409 (13-1854) days (dogs, n = 78) and 395 (28-1670) days (cats, n = 21). During follow-up, neoplasia was diagnosed in 4/27 hypercobalaminemic dogs versus 3/51 control dogs (P = 0.23) and 1/8 hypercobalaminemic cats versus 0/13 control cats (P = 0.38). Pancreatitis was more frequent in hypercobalaminemic dogs (P = 0.006). Hypercobalaminemic dogs had higher serum total protein (P = 0.014), globulin (P = 0.001), and CRP (P = 0.032) concentrations and lower serum sodium (P = 0.012) and chloride (P = 0.033) concentrations than controls. Hypercobalaminemic cats had higher serum total protein concentrations than controls (P = 0.008). CONCLUSION: Our results suggest that hypercobalaminemia is not associated with the presence or development of neoplasia in dogs and cats but may be associated with systemic inflammatory conditions, including pancreatitis, in dogs.

The ultrasonographic appearance of renal medullary striations and their association with renal disease and renal histopathology in domestic cats.

Medullary striations (MS) have been anecdotally observed on ultrasound of feline kidneys; however, their significance is unknown. Aims of this retrospective, case control, pilot study were to describe the appearance, prevalence, and clinicopathological correlates of MS in a referral feline population. Still images from 1247 feline abdominal ultrasound studies performed between 2011 and 2021 were reviewed. Cats with MS were identified and compared with age-matched controls. Serum urea, creatinine, calcium, phosphate, and calcium-phosphate-product, plus urine specific gravity, urine protein: creatinine ratio (UPC), prevalence of active sediment (defined as > 5 red (RBC) or white blood cells (WBC) per high-power field) and prevalence of positive urine culture were compared between MS and control groups using the Mann-Whitney U test or Fisher's Exact test. Data are presented as median [range]. 27 cats were identified as having MS, giving a prevalence of 2.2% with a significantly higher proportion being seen in males (P = 0.018). Medullary striation cats had significantly higher UPC values than controls (0.46 [0.16-7.57] vs. 0.16 [0.07-2.27]; P = 0.006). Cats with MS were more likely to have active urinary sediments (39% vs 8%, P = 0.023), but no difference in prevalence of positive urinary cultures was observed between groups. There was no significant difference in other parameters between MS and control cats. Renal histopathology performed in three MS cats revealed focal regions of linear medullary fibrosis. Medullary striations are associated with proteinuria and urinary tract inflammation in cats, which may reflect renal tubular dysfunction and/or inflammation. Hence identification might allow for earlier detection of renal pathology.

Measuring disability in amyotrophic lateral sclerosis/motor neuron disease: the WHODAS 2.0-36, WHODAS 2.0-32, and WHODAS 2.0-12.

Aim: To investigate whether the World Health Organization Disability Assessment Schedule 2.0 (WHODAS) can provide interval level measurement of disability in Amyotrophic Lateral Sclerosis (ALS), allowing parametric analyses. Methods: Data on the WHODAS 12, 32, and 36-item versions, from 1120 patients studied at one or more time points, were fit to the Rasch model and comparisons made against ALSFRS-R, King's staging, and mortality. Trajectory modeling was undertaken for a newly diagnosed (≤6 months) cohort of 454 individuals. Results: Total scores for WHODAS 32 and 36-item versions can be converted to interval level measurement suitable for individual clinical use, and the 12-item WHODAS total for group use. The 36-item version is shown to be equivalent to the 32-item version. Expected correlations were seen with King's staging, ALSFRS-R, and EQ-5D-5L. Trajectory analysis of disability (WHODAS 2.0) showed three clearly demarcated groups with differences in King's staging, depressive symptomatology and mortality, but not age. Conclusions: The WHODAS 2.0 is a brief patient reported outcome measure which can be used to measure disability in ALS. Provided the patient answers all 36 (32 if not working) items, the conversion table produces an interval level estimate for parametric analyses. The different trajectories demonstrated from diagnosis support the concept of a prodromal period, and suggest the WHODAS 2.0 could be used for surveillance of at risk populations, such as those with genetic predisposition.

Serum amyloid A and other clinicopathological variables in cats with intermediate- and large-cell lymphoma.

OBJECTIVES: Serum amyloid A (SAA) concentrations are increased in cats with lymphoma vs healthy cats; however, the association between SAA concentrations and prognosis in cats with lymphoma is unclear. The aim of this study was to evaluate if SAA concentrations were different in cats with nasal vs non-nasal lymphoma, if SAA concentrations are prognostic in patients treated with high-dose chemotherapy and if SAA concentrations are correlated with other clinicopathological variables. METHODS: Cats diagnosed with intermediate- or large-cell lymphoma between 2012 and 2022 with SAA concentration data available were included. Associations between tumour site (nasal vs non-nasal), stage, response to treatment and SAA concentration were evaluated using non-parametric statistics. Associations between SAA concentrations and stage with survival time were evaluated using Cox regression analysis. Patients with nasal tumours and those not receiving high-dose chemotherapy were excluded from the survival analyses. RESULTS: Thirty-nine cats were included. Median SAA concentrations were significantly higher in non-nasal compared with nasal lymphoma (42 µg/ml [range <0.3-797] vs <0.3 µg/ml [range <0.3-0.9]; P = 0.026). SAA concentrations did not correlate with tumour stage. Median survival time for patients with non-nasal tumour and undergoing chemotherapy was 49 days (range 2-1726). Responders had a better median survival time than non-responders (273 days [range 43-1728] vs 39 days [range 2-169]; P <0.001), whereas SAA concentrations were not associated with survival time. Lower haematocrit at presentation was associated with a reduced median survival time (P = 0.007). CONCLUSIONS AND RELEVANCE: In the population examined, no correlation between serum concentration of SAA and prognosis in patients with lymphoma was identified, while low haematocrit and lack of response to treatment were both found to be associated with survival time. SAA concentrations were elevated in patients with non-nasal lymphoma vs patients with tumours confined to the nasal cavity.

Parasitic helminths and the host microbiome - a missing 'extracellular vesicle-sized' link?

Infections by gastrointestinal (GI) helminths have been associated with significant alterations of the structure of microbial communities inhabiting the host gut. However, current understanding of the biological mechanisms that regulate these relationships is still lacking. We propose that helminth-derived extracellular vesicles (EVs) likely represent key players in helminth-microbiota crosstalk. Here, we explore knowledge of helminth EVs with an emphasis on their putative antimicrobial properties, and we argue that (i) an enhanced understanding of the mechanisms governing such interactions might assist the discovery and development of novel strategies of parasite control, and that (ii) the identification and characterisation of helminth molecules with antimicrobial properties might pave the way towards the discovery of novel antibiotics, thus aiding the global fight against antimicrobial resistance.

Retrospective analysis of association between hepatopathy and serum DGGR lipase activity in dogs: a pilot study.

1,2-o-dilauryl-rac-glycero-3-glutaric acid-(6'-methylresorufin) ester (DGGR) lipase assays are used to measure lipase activity in the diagnosis of pancreatitis. The effect of hepatic lipases released from damaged hepatocytes on serum DGGR lipase activity has not been reported, to our knowledge. We identified dogs with histologically confirmed liver lesions and concurrent unremarkable pancreatic histology, and dogs with no histologic evidence of hepatic or pancreatic disease. Dogs with relevant comorbidities were excluded. The hepatopathy group (n = 7) included 4 dogs with inflammatory hepatopathies, 2 with hepatic neoplasia, and 1 with unspecified (non-inflammatory, non-neoplastic) hepatopathy. The control group (n = 5) included one dog each with enteritis, subcutaneous hemangiosarcoma, hydrocephalus, myelomalacia, and tetanus. A Mann-Whitney U test compared selected biochemical parameters including serum DGGR lipase, alkaline phosphatase, alanine aminotransferase, and amylase activities, with statistical significance defined as p ≤ 0.05. Data are presented as median and range. Serum DGGR lipase activity (RI: <44 IU/L) was not different between the hepatopathy (52 IU/L; range: 27-85 IU/L) and control (37 IU/L, 25-105 IU/L; p = 0.947) groups. Serum amylase activity (RI: 256-1,610 IU/L) was significantly higher in the hepatopathy group (830 IU/L; 711-1,210 IU/L) than the control group (541 IU/L, 336-695 IU/L; p = 0.028). No association or correlation between serum DGGR lipase activity and hepatic lesions (based on histologic or biochemical findings) was identified, suggesting that clinically relevant changes in serum DGGR lipase activity may not be expected secondary to hepatopathy alone.

Electrodiagnostic findings in facial onset sensory motor neuronopathy (FOSMN).

OBJECTIVE: To determine the electrodiagnostic characteristics of facial onset sensory and motor neuronopathy (FOSMN). METHODS: Electrophysiological data from 10 FOSMN patients in Newcastle-upon-Tyne and Sydney were reviewed. Relevant literature was reviewed. RESULTS: Findings on standard electrophysiological assessment were in broad agreement with those published: blink reflexes were abnormal in all but one patient; sensory nerve action potentials were reduced but compound muscle action potentials preserved; mixed acute and chronic neurogenic change was identified on needle electromyography in bulbar and cervico-thoracic muscles in approximately 50% of patients. Upper limb somatosensory evoked potential (SEP) central conduction times were increased (n = 4) and progressed on repeat testing (n = 3). Upper motor neuron dysfunction was revealed by several measures [ipsilateral motor evoked potentials (MEPs) (n = 1); reduced short interval intra-cortical inhibition on threshold-tracking transcranial magnetic stimulation (n = 2); absent beta-band intermuscular coherence (n = 3)]. CONCLUSIONS: Electrodiagnostic investigation of FOSMN should include blink reflex testing, SEPs and tests of upper motor neuron function. The combination of progressive lower motor neuron disease and upper motor neuron disease on neurophysiological investigation provides further support for the contention that FOSMN is a rare variant of motor neurone disease. SIGNIFICANCE: These findings will aid the neurologist and neurophysiologist in making a confident diagnosis of FOSMN, thus expediting appropriate care.

Can the neutrophil count from hepatic fine-needle aspirate cytology be used to diagnose hepatitis in dogs? A pilot study.

BACKGROUND: The cytologic diagnosis of inflammation on canine hepatic aspirates can be confounded by neutrophilic infiltrates in the liver of dogs with nodular regeneration, by extramedullary hematopoiesis, and by marked blood contamination. OBJECTIVES: We aimed to assess the association between neutrophil counts on hepatic cytology and the histopathologic diagnosis in dogs with hepatitis and non-inflammatory hepatopathy. We also sought to determine a cut-off value for the cytologic diagnosis of hepatitis. METHODS: In a retrospective blinded pilot study, three observers independently reviewed hepatic aspirates that had corresponding histopathologic examinations performed within 2 days. The number of neutrophils per 200 hepatocytes was determined and averaged among observers. Only neutrophils within or directly in contact with a cluster of ≥5 hepatocytes were counted, and only intact hepatocytes within an approximate monolayer were included. Data are presented as the median (range), and the Mann-Whitney U test is used to make comparisons between groups. RESULTS: Eighteen cases were included (13 hepatitis and five vacuolar hepatopathy). Aspirates with a histopathologic diagnosis of hepatitis had increased numbers of neutrophils compared with those of vacuolar hepatopathy (7.7 [0.3-18.3] vs 3.0 [1.0-5.3]; P = .038). Receiver operating characteristic curve analysis indicated that ≥6 neutrophils were 61.5% (CI 31.6%-86.1%) sensitive and 100% (CI 47.8%-100%) specific for identifying hepatitis. CONCLUSIONS: Liver aspirates from hepatitis cases have a higher number of neutrophils on cytology compared with those from vacuolar hepatopathy; however, larger studies, including those with dogs with other liver pathologies, are required. Identification of six or more neutrophils per 200 hepatocytes is highly suggestive of hepatitis.

Stomatocytosis in a Beagle and Australian Cattle Dog.

BACKGROUND: Canine stomatocytosis is a well-recognized rare erythrocyte disorder characterized by nonsyndromic forms with selective erythroid involvement, syndromic forms with extra-hematologic disease, and acquired forms. OBJECTIVES: We describe serial clinicopathologic changes in two dogs with stomatocytosis of breeds that are different from those previously reported. METHODS: Blood samples were obtained from a 12-year-old female neutered Australian Cattle Dog and a 12-year-old male neutered Beagle for hematologic and biochemical analyses, including a morphologic examination of peripheral blood films. Serial clinicopathologic data were reviewed, including CBCs performed by the referring veterinary surgeons. RESULTS: Serial CBC data in both cases reported a variable decrease in RBC numbers commonly associated with a normal hematocrit, macrocytosis, hypochromasia, changes in red cell distribution width parameters including marked histogram abnormalities in volume distribution of the RBC population, and mildly increased or normal reticulocyte counts. Morphologic examination of peripheral blood films identified variable numbers of stomatocytes, knizocytes (Case 1, Day 1, Day 4), mild anisocytosis, mild macrocytosis, and mild polychromasia. CONCLUSIONS: In both cases, the changes exhibited in the erythrogram raise suspicion for an RBC membrane disorder with cell volume dysregulation and stomatocytosis, although they did not appear to cause clinically relevant hemolysis.

Size-Exclusion Chromatography Separation Reveals That Vesicular and Non-Vesicular Small RNA Profiles Differ in Cell Free Urine.

Urinary extracellular vesicles (EVs) and their RNA cargo are a novel source of biomarkers for various diseases. We aimed to identify the optimal method for isolating small (<200 nm) EVs from human urine prior to small RNA analysis. EVs from filtered healthy volunteer urine were concentrated using three methods: ultracentrifugation (UC); a precipitation-based kit (PR); and ultrafiltration (UF). EVs were further purified by size-exclusion chromatography (SEC). EV preparations were analysed with transmission electron microscopy (TEM), Western blotting, nanoparticle tracking analysis (NTA) and an Agilent Bioanalyzer Small RNA kit. UF yielded the highest number of particles both before and after SEC. Small RNA analysis from UF-concentrated urine identified two major peaks at 10-40 nucleotides (nt) and 40-80 nt. In contrast, EV preparations obtained after UC, PR or SEC combined with any concentrating method, contained predominantly 40-80 nt sized small RNA. Protein fractions from UF+SEC contained small RNA of 10-40 nt in size (consistent with miRNAs). These data indicate that most of the microRNA-sized RNAs in filtered urine are not associated with small-sized EVs, and highlights the importance of removing non-vesicular proteins and RNA from urine EV preparations prior to small RNA analysis.

Resting and postprandial serum bile acid concentrations in dogs with liver disease.

BACKGROUND: Serum bile acids (SBAs) are frequently measured in dogs. However, there is limited data comparing SBAs in different liver diseases diagnosed according to standardized histological criteria. OBJECTIVES: To compare resting and postprandial SBAs, and determine their sensitivity and specificity, for various liver diseases in dogs. ANIMALS: Three hundred and forty-one client-owned dogs with suspected liver disease that had a liver biopsy and SBAs measured. METHODS: Multicenter retrospective study. Cases were classified according to standardized histological criteria. The sensitivity and specificity of resting and postprandial SBAs for the diagnosis of each liver disease, and all liver diseases combined, were calculated. RESULTS: The median resting SBAs were highest in dogs with cirrhosis (98.8 µmol/L; range, 6-135) and congenital circulatory anomalies (CCa; 79.45 µmol/L; 0.3-705). The highest median postprandial concentrations were found in CCa (126 µmol/L; 0-726) and chronic hepatitis (CH; 54.3 µmol/L; 0-260). Using the cut-off value of 10 µmol/L, the highest sensitivities of resting SBAs were recorded in dogs with CCa (87.5%; 95% confidence interval, 76.8-94.4) and CH (81.1%; 71.5-88.6). The sensitivities of postprandial SBAs were the highest in cholangitis (100%; 47.8-100.0) and CCa (91.1%; 78.8-97.5). The specificities of resting and postprandial SBAs for all diseases were 49.3% (37.6-61.1) and 29.7% (15.9-47.0), respectively. CONCLUSIONS AND CLINICAL IMPORTANCE: Postprandial SBAs are more sensitive but less specific than resting SBAs for the diagnosis of liver disease. There were dogs in all categories of liver disease with resting SBAs <10 and >90 µmol/L. Therefore, careful interpretation of both normal and elevated values is required.

Correlation between cytologic features and histologic grades in cutaneous and subcutaneous soft tissue sarcomas in dogs-A pilot study.

Currently, canine soft tissue sarcoma (STS) grading is based on histopathology. In humans, several studies have demonstrated concordance between cytologic grading systems for STS and histologic grade. The aim of this study was to correlate several cytologic parameters (smear cellularity, anisokaryosis, nucleolar malignancy score, multinucleation, and the number of mitotic figures per 200 cells) that form part of a human STS cytologic grading system, with histologic grades of canine cutaneous and subcutaneous STS. Three observers (blinded) reviewed the cytologic preparations independently from cases with confirmed histologic diagnoses of STS. A cytologic grading score was assigned for each parameter. Correlations between cytologic grading scores (averaged between observers) and histologic grades were assessed using Spearman's correlation coefficient, with statistical significance defined as P < .05. Twenty-one cases were included in the study (10 Grade I STS, nine Grade II STS, and two Grade III STS). The number of mitotic figures (≥3) per 200 cells was the only parameter that showed a significant but weak, positive correlation with histologic grade (rs  = .469; P = .032). No Grade I tumors had ≥3 mitotic figures per 200 cells; however, ≥3 mitotic figures per 200 cells were only observed in 33% of Grade II tumors and 50% (one out of two) of the Grade III tumors. This pilot study suggests that an increased number of mitotic figures seen on cytology might correlate with higher grade STS; however, the sensitivity of this parameter for grading STS appears to be low.

Preliminary evaluation of a salivary urea test strip method for use in dogs.

BACKGROUND: Salivary urea concentrations correlate with serum urea concentrations in dogs and humans. Salivary urea concentrations can now be determined semi-quantitatively using a salivary urea test strip method that has been validated for use in humans. OBJECTIVES: We aimed to evaluate the repeatability of the salivary urea test strip score, and the correlation between the salivary urea test strip scores and serum urea concentrations in dogs. METHODS: Intra-run and inter-run variabilities were determined (n = 10 in triplicate). Correlations between salivary urea test strip scores and serum urea concentrations in dogs were assessed using the Spearman's correlation coefficient. Receiver operator curve analysis was used to evaluate the diagnostic performance of the salivary urea test strip score to identify dogs with serum urea concentration >7.4 mmol/L (upper limit of laboratory RI). RESULTS: The intra-run repeatability was good (28/30 concordant results) whereas the inter-run repeatability was moderate (23/30 concordant results). Salivary and serum urea concentrations showed a moderately positive correlation (rs  = .63, n = 33; P < .0001). A salivary urea test strip score ≥4 was 57% sensitive and 96% specific for detecting a serum urea concentration >7.4 mmol/L. CONCLUSIONS: Uremia can be detected using salivary urea test strips in dogs. Based on our preliminary data, salivary urea test strip scores of 1 or 2 might exclude clinically relevant uremia in most cases; however, it is recommended that the salivary urea test be repeated in dogs with a test strip score of 3. Dogs with a salivary urea test strip score of ≥4 would likely require additional investigations.

Hepatocyte expression and prognostic importance of senescence marker p21 in liver histopathology samples from dogs with chronic hepatitis.

BACKGROUND: Chronic hepatitis (CH) occurs commonly in dogs but is associated with a variable and largely unpredictable prognosis. p21, a cell-cycle inhibitor and marker of cellular senescence, is upregulated in human liver disease and is a better prognostic marker than histological or clinical scoring systems. OBJECTIVE: To quantify hepatocyte p21 immunopositivity in histopathology samples from dogs with CH and determine its association with outcome. ANIMALS: Twenty-six client-owned dogs with histologically confirmed CH, and 15 dogs with normal liver histology. METHODS: Medical records and liver histopathology samples were retrospectively reviewed to identify cases of CH. Immunohistochemistry for p21 was performed on all samples and hepatocyte immunopositivity was visually quantified. Relationships between p21 and dog age and dog survival time were statistically evaluated. RESULTS: Hepatocyte p21 immunopositivity in dogs with CH was high (median percentage of positive hepatocytes: 90%, range: 20%-98%) and exceeded 70% in 23/26 cases with no association with age. In control dogs, p21 immunopositivity was low (≤15% positive hepatocytes in 12/15 cases) and was positively correlated with age (rs = 0.63; P = .011). Dogs with p21 immunopositivity exceeding 91.8% (upper tercile) had significantly shorter survival compared to dogs with less than 88.9% immunopositivity (lowest tercile; 218 versus 874 days, P = .006). Increasing hepatocyte p21 immunopositivity was significantly negatively associated with survival time (HR 4.12; 95% CI 1.34-12.63; P = .013). CONCLUSIONS AND CLINICAL IMPORTANCE: Marked p21 immunopositivity in dogs with CH might be indicative of widespread hepatocellular senescence. A significant association with survival time also suggests a potential value for p21 quantification in determining prognosis.

Validation of a commercial 1,2-o-dilauryl-rac-glycero glutaric acid-(6'-methylresorufin) ester lipase assay for diagnosis of canine pancreatitis.

The objectives of this study were fourfold: technical validation of a commercial canine 1,2-o-dilauryl-rac-glycero glutaric acid-(6'-methylresorufin) ester (DGGR) lipase assay, to calculate a reference interval for DGGR lipase by the indirect a posteriori method, to establish biological validity of the assay, and to assess agreement between DGGR lipase and specific canine pancreatic lipase (Spec cPL) assays. Dogs with histologically confirmed acute pancreatitis (n=3), chronic pancreatitis (n=8) and normal pancreatic tissue (n=7) with stored (-80°C) serum samples were identified. Relevant controls were selected. Precision, reproducibility and linearity of DGGR lipase, and the effect of sample haemolysis and freezing, were assessed. Sensitivity and specificity of DGGR lipase and Spec cPL were determined. Agreement between these two parameters was calculated using Cohen's kappa coefficient (κ). The DGGR lipase assay demonstrated excellent precision, reproducibility and linearity. Sample haemolysis and storage at -80°C for 12 months did not influence the assay. DGGR lipase (>245IU/l) and Spec cPL (>400µg/l) both showed poor sensitivity but excellent specificity for acute pancreatitis, and poor to moderate sensitivity but excellent specificity for chronic pancreatitis. Substantial agreement (κ=0.679) was found between DGGR lipase and Spec cPL. The validated DGGR lipase assay had similar sensitivity and specificity for the diagnosis of acute and chronic pancreatitis to Spec cPL. DGGR lipase is a reliable alternative to Spec cPL for the diagnosis of pancreatitis.

Detection and seroprevalence of morbillivirus and other paramyxoviruses in geriatric cats with and without evidence of azotemic chronic kidney disease.

BACKGROUND: Feline morbillivirus (FeMV) is associated with the presence of tubulo-interstitial nephritis (TIN) in cats, however the seroprevalence of FeMV in the UK and the association between the presence of FeMV and renal azotemia is unknown HYPOTHESIS/OBJECTIVES: To identify whether paramyxoviruses are present in urine samples of geriatric cats and to develop an assay to assess FeMV seroprevalence. To investigate the relationship between both urinary paramyxovirus (including FeMV) excretion and FeMV seroprevalence and azotemic chronic kidney disease (CKD). ANIMALS: Seventy-nine cats (40 for FeMV detection; 72 for seroprevalence). METHODS: Retrospective cross-sectional, case control study. Viral RNA was extracted from urine for RT-PCR. PCR products were sequenced for virus identification and comparison. The FeMV N protein gene was cloned and partially purified for use as an antigen to screen cat sera for anti-FeMV antibodies by Western Blot. RESULTS: Feline morbillivirus RNA from five distinct morbilliviruses were identified. Detection was not significantly different between azotemic CKD (1/16) and nonazotemic groups (4/24; P = .36). Three distinct, non-FeMV paramyxoviruses were present in the nonazotemic group but their absence from the azotemic group was not statistically significant (P = .15). 6/14 (43%) azotemic cats and 40/55 (73%) nonazotemic cats were seropositive (P = .06). CONCLUSIONS AND CLINICAL IMPORTANCE: Feline morbillivirus was detected in cats in the UK for the First time. However, there was no association between virus prevalence or seropositivity and azotemic CKD. These data do not support the hypothesis that FeMV infection is associated with the development of azotemic CKD in cats in the UK.

Investigation of the association between serum protein concentrations and concurrent chronic kidney disease in hyperthyroid cats.

Our objective was to identify if changes in serum protein concentrations occur in hyperthyroidism and to assess their association with the development of azotaemia following treatment. Initially non-azotaemic hyperthyroid cats and healthy older cats were included. Serum concentrations of protein fractions were determined by agarose gel electrophoresis and compared between; hyperthyroid and control cats, initially non-azotaemic hyperthyroid cats which developed azotaemia in a 4month follow up period (masked-azotaemic) and those which remained non-azotaemic, and hyperthyroid cats before and at the time of restoration of euthyroidism. Data are presented as median [25th, 75th percentiles]. Hyperthyroid cats (n=56) had higher serum α2 globulin concentrations (12.5 [10.9, 13.1] g/L vs. 9.8 [3.0, 11.4] g/L; P<0.001) and lower serum γ globulin concentrations (11.4 [9.1, 13.3] g/L vs. 14.0 [12.4, 16.8] g/L; P=0.001) than control cats (n=26). Following treatment, serum total globulin concentration increased (from 38.6 [35.4, 42.8] g/L to 42.3 [39.0, 45.7] g/L; P<0.001), serum α2 globulin concentration decreased (from 12.5 [10.9, 13.9] g/L to 11.5 [10.1, 12.6] g/L; P<0.001) and serum γ globulin concentration increased (from 11.4 [9.0, 13.3] g/L to 14.0 [12.4, 16.8] g/L; P<0.001). Serum concentrations of total globulin or globulin fractions were not significantly different between masked-azotaemic and non azotaemic groups. In conclusion, hyperthyroidism is associated with altered serum concentrations of the α2 and γ globulin fractions, however these changes were not associated with the development of azotaemic chronic kidney disease following treatment.

Effect of prewarming EDTA blood samples to 37°C on platelet count measured by Sysmex XT-2000iV in dogs, cats, and horses.

BACKGROUND: Pseudothrombocytopenia secondary to platelet clumping is a common cause of preanalytic error for platelet counts in dogs, cats, and horses. In human beings, it is suggested that prewarming blood samples to 37°C prior to hematology analysis will reduce platelet clumping. OBJECTIVES: The purpose of the study was to evaluate the effect of prewarming EDTA blood samples to 37°C on measured platelet counts and other hematologic variables. METHODS: The EDTA blood samples from dogs, cats and horses submitted to the clinical pathology laboratory at the University of Cambridge were included. Complete blood cell counts performed using a Sysmex XT-2000iV hematology analyzer were done on samples at room temperature (approximately 22°C) and following warming of the samples to 37°C in a water bath. The Wilcoxon signed rank test was used to compare hematologic variables, including platelet count, before and after sample warming to 37°C. Data are presented as median (25(th) , 75(th) percentile) increase. RESULTS: Blood samples from 39 dogs, 19 cats, and 10 horses were included. Sample warming to 37°C resulted in a statistically significant increase in platelet counts in dogs (11 [-2, 30] ×10(9) /L), cats (36 [14, 84] ×10(9) /L), and horses (42 [31, 79] ×10(9) /L). Sample warming did not significantly affect other hematologic variables. CONCLUSIONS: Prewarming EDTA blood samples to 37°C prior to hematologic analysis increased platelet counts overall in canine, feline, and equine blood, but did not abrogate platelet clumping and pseudothrombocytopenia fully in some cases. Furthermore, true pseudothrombocytopenia was not confirmed in these animals.