ORCID Profile
0000-0003-4879-9504
Current Organisation
Murdoch University
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Publisher: Wiley
Date: 06-11-2013
DOI: 10.1111/VCP.12091
Publisher: Elsevier BV
Date: 04-2017
DOI: 10.1016/J.RVSC.2017.02.007
Abstract: High density lipoproteins (HDLs) are pivotal in innate immunity and decrease in serum during inflammation. Several studies have been done about lipoprotein changes in transition cows but little is known about their changes in newborn calves. The aim of this study is to provide information about HDLs in newborn calves, by defining the possible age-related changes in healthy calves compared with adults and by assessing the possible differences in calves with inflammation. Lipoprotein electrophoretic separation (reported as percentages) and colorimetric measurement of HDL (HDL-C) were performed on healthy cows and calves in order to identify possible differences in the lipoprotein profile due to the age. Then, age-matched calves with inflammatory conditions were also evaluated. Results showed that in calves HDL% and VLDL% were lower (mean values±SD: 77.6%±8.6% and 2.6%±2.5%, respectively) and LDL% was higher (19.7%±7.4%) than in adults (89.0%±3.9% 5.2±2.1% and 5.8%±3.1%, respectively). Sick calves revealed a decrease of both HDL% (mean values ± SD: 61.0%±22.1%) and HDL-C (22.8±11.6mg/dL) and an increase of VLDL% (12.1% ±13.1%) compared with controls (77.6%±8.6% 41.5±11.2mg/dL and 2.6%±2.5%, respectively). Paraoxonase-1 activity, influenced by inflammation and oxidation, was measured, and it appeared correlated with HDL% and HDL-C in sick calves. In conclusion, this study revealed that HDLs concentration in healthy calves is lower than in adults, and further decreases in calves with inflammation, likely due to oxidation.
Publisher: SAGE Publications
Date: 08-2013
Abstract: Many feline breeds have been generated from a small number of ancestors. Thus, breed-specific peculiarities can be expected, which could include haematological and biochemical measurements. Despite this, there are only a few reports on breed-specific reference intervals (RI). This information is essential in routine practice where results from in idual patients are usually compared with an RI. The aim was to compare haematological and biochemical data from clinically healthy Abyssinian, Holy Birman, Norwegian Forest and Siberian cats with published RIs to assess whether the published RIs are acceptable in these breeds. Comparison with established RIs using guidelines from the National Committee for Clinical Laboratory Standards and the American Society of Veterinary Clinical Pathology, revealed a number of breed-related clinicopathological differences. New RIs were established, but in most cases the new RIs overlapped with published RIs, and the use of the breed-specific data would minimally affect the clinical interpretation of laboratory results. Important differences that could result in misinterpretation of laboratory results were as follows: microcytosis and high α 2 -globulin concentrations in Abyssinian cats high serum creatinine, α 2 -globulin and glucose concentrations in Holy Birman cats high serum alkaline phosphatase activity and calcium and phosphate concentration in Norwegian Forest cats low β 2 -globulin and γ-globulin concentrations in Norwegian Forest and Siberian cats. Breed-specific RIs should be used for these analytes. In addition, care should be taken in interpreting clinicopathological data in purebred cats for which specific RIs have not been established.
Publisher: Wiley
Date: 12-2006
Publisher: MDPI AG
Date: 17-10-2023
Publisher: Wiley
Date: 12-2019
DOI: 10.1111/VCP.12806
Abstract: Synthetic colloid solutions, administered by rapid infusion to volume-depleted dogs, might be present in high concentrations in subsequent urine s les. The potential for these solutions to affect the performance of ELISA measurements due to s le matrix effects when studying kidney injury biomarkers requires investigation. We aimed to investigate two different synthetic colloid solutions, 4% succinylated bovine gelatin (GEL) and 6% hydroxyethyl starch (HES), for potential interferences with a commercially available canine neutrophil gelatinase-associated lipocalin (NGAL) ELISA. Assay interference was assessed by measuring the linearity of NGAL concentrations measured using a canine NGAL ELISA after serial dilution of a canine pooled urine s le with an assay diluent, GEL, or HES. NGAL recovery from urine specimens containing up to 75% HES and up to 62.5% GEL was within acceptable limits (80%-120%). NGAL recovery from the urine specimen containing 75% GEL was poor (76%). Linear regression analysis demonstrated excellent linearity under dilution when a canine urine s le was diluted with the assay diluent, GEL, or HES. The presence of large amounts (>62.5%) of GEL in canine urine s les could cause negative interference in the performance of the NGAL ELISA investigated.
Publisher: Wiley
Date: 22-08-2018
DOI: 10.1111/VCP.12646
Abstract: Proteinuria quantification with the urinary protein-to-creatinine (UPC) ratio is part of the diagnostic process in feline patients suspected of chronic kidney disease (CKD). In affected cats, monitoring and substaging of the UPC according to the International Renal Interest Society (IRIS) guidelines is also necessary for appropriate patient management. No information is available about the possible effects of analytical variability on urinary proteins (UPs) and the UPC ratio in cats. This study aimed to determine whether imprecision and method-dependent differences due to the two dye-binding methods, pyrogallol red-molybdate (PRM) and Coomassie brilliant blue (CBB), could affect IRIS substaging. Urine s les were collected from proteinuric and nonproteinuric cats. Intra-assay and inter-assay repeatability were assessed with both the PRM and CBB methods. Urinary supernatants (n = 120) were tested using both methods. Agreement between the methods and concordance with s le classification according to IRIS guidelines were determined. On average, the PRM method yielded a higher CV (UP 8.4 ± 5.2% UPC 9.5 ± 4.8%) than the CBB method (UP 5.6 ± 2.6% UPC 7.2 ± 2.6%), but similar rates of misclassification were found in s les with UPC ratios close to the IRIS cut-off. Although the two methods were correlated, the CBB method tended to yield UPs and UPC ratios that were significantly higher (P < 0.0001) than that of the PRM method. The Passing-Bablok test also found constant and proportional errors between the PRM and CBB methods. Concordance in substaging s les according to IRIS was good (k coefficient = 0.62). The two methods were precise, but the higher UPC ratios obtained with the CBB methods might affect the interpretation using the IRIS guidelines and clinical decisions.
Publisher: Wiley
Date: 27-10-2014
DOI: 10.1111/VCP.12207
Publisher: Wiley
Date: 17-08-2018
DOI: 10.1111/VCP.12647
Abstract: High molecular weight (MW) synthetic colloids interfere with refractometry. The effect of low MW synthetic colloids on refractometry is unknown. To investigate the effect of 6% hydroxyethyl starch 130/0.4 (HES) and 4% succinylated gelatin (GELO) on the refractometric measurement of total plasma protein (TPP) and colloid osmotic pressure (COP) in canine plasma. Heparinized plasma from 10 dogs was diluted with 0.9% saline (NaCl), HES, and GELO in ratios of plasma:fluid, 9:1, 8:2, 7:3, 6:4, and 5:5. The biuret TPP (TPPb), refractometric TPP (TPPr), and COP assays were performed for all dilutions. Agreement (bias, 95% limits of agreement (LOA)) was tested between the TPPb (NaCl) and the TPPr of the NaCl, HES, and GELO plasma dilutions, and between the TPPb (NaCl) and the TPPb of the HES and GELO plasma dilutions. A small bias (0.06 g/dL, 95% LOA -0.51 to 0.39 g/dL) existed between TPPb (NaCl) and TPPr (NaCl). A large bias was found between TPPb (NaCl) and TPPr (HES) (1.38 g/dL, 95% LOA 0.30 to 3.10 g/dL), and TPPr (GELO) (1.18 g/dL, 95% LOA 0.53 to 2.88 g/dL). A small bias was found between TPPb (NaCl) and TPPb (HES) (0.08 g/dL, 95% LOA -2.04 to 2.20 g/dL), and a large bias was found between TPPb (NaCl) and TPPb (GELO) (1.00 g/dL, 95% LOA -0.59 to 2.60 g/dL). Increasing overestimation of TPPb (NaCl) by TPPr (HES and GELO) and TPPb (GELO) occurred with increasing colloid volumes. HES and GELO interfered with TPPr, and GELO also interfered with TPPb, in vitro. These variables should be interpreted with caution for dogs that have received these synthetic colloids.
Publisher: Wiley
Date: 12-2010
Publisher: Wiley
Date: 12-2020
DOI: 10.1111/VCP.12923
Publisher: Wiley
Date: 03-2014
DOI: 10.1111/VCP.12121
Abstract: Information about laboratory reference intervals (RIs) of European Hedgehog (Erinaceus europaeus) hospitalized at rehabilitation centers is scarce. The purpose of this study was to establish hematologic and biochemical RIs for rehabilitated hedgehogs before the release into the wild, and to assess whether sex and management of the center influence laboratory results. Blood was collected from 50 hedgehogs at 3 centers. Thirty-eight animals were included in the study based on normal body weight, absence of clinical signs of disease, Bunnell index > 0.80, and absence of hibernation during overwintering. CBCs were performed using an automated laser cell counter followed by morphologic analysis of blood smears. Clinical biochemistry was performed using an automated spectrophotometer. RIs were determined as recommended by the ASVCP guidelines. Hematology profiles revealed a prevalence of lymphocytes, a constant presence of nucleated RBCs, Howell-Jolly bodies and basophils, and bilobed nuclei in neutrophils and eosinophils. Biochemistry profiles were characterized by higher creatinine and urea concentrations, and higher ALP and GGT activities compared with other domestic species. The sex did not influence the results. Conversely, numbers of eosinophils, activated and large granular lymphocytes, and concentrations of total protein, glucose and cholesterol were different among the centers, likely due to different management practices (eg, antiparasitic treatments, environmental exposure to microorganisms, diet). The RIs established in this study can be used to monitor the health status of hedgehogs in rehabilitation centers. As management practices appeared to influence some variables, it is recommended to standardize the management protocols to minimize their influence on laboratory data.
Publisher: Wiley
Date: 07-09-2012
Publisher: Wiley
Date: 02-04-2015
DOI: 10.1111/VCP.12241
Publisher: SAGE Publications
Date: 22-11-2020
Abstract: Paraoxonase-1 (PON-1) activity is a new inflammatory and oxidative marker. Technical effects and biological factors could affect the accuracy of PON-1 activity measurement. We investigated the effects of storage at different temperatures, repeated freeze–thaw cycles, interferences from hemolytic, lipemic, and icteric s les, and seasonal effects on PON-1 activity in horses. We evaluated 2 substrates with an automated spectrophotometer. Ten equine serum s les were stored under different conditions. Although storage at room (21°C) or refrigeration (4°C) temperature induced a statistically significant decrease ( p 0.05) in PON-1 activity, this is not diagnostically relevant. PON-1 activity in frozen s les (−20°C) was stable for short-term storage diagnostically significant ( p 0.01) fluctuations were observed after 1 mo. Four repeated freeze–thaw cycles were assessed, and all cycles affected PON-1 activity ( p 0.01) however, this was diagnostically significant only after the 4th cycle. Hemolysis induced an overestimation of PON-1 activity lipemia and hyperbilirubinemia did not change PON-1 activity. Thirty-four horses were s led monthly for 1 y, and PON-1 activity was higher in autumn ( p 0.05) and winter ( p 0.05) than in spring and summer.
Publisher: Wiley
Date: 06-2010
Publisher: Informa UK Limited
Date: 29-01-2020
Publisher: SAGE Publications
Date: 09-2008
DOI: 10.1177/104063870802000520
Abstract: In humans, homocysteine (Hcy) is employed to monitor renal, cardiovascular, and other diseases and their complications. The aim of the current study was to define the analytical performances of an enzymatic method not yet validated in dogs for measuring homocysteine, and to assess the possible clinical usefulness of Hcy measurement. Using conventional approaches, the analytical performances were investigated by assessing, imprecision, inaccuracy, and interference of hemoglobin, triglycerides, and bilirubin. The possible clinical usefulness of Hcy determination was assessed by comparing the results of healthy dogs ( n = 8) with those of dogs with heart disease ( n = 10), inflammation ( n = 6), gastrointestinal disorders ( n = 7), neoplasia ( n = 8), renal failure ( n = 4), trauma ( n = 7), and other miscellaneous diseases ( n = 6). Preliminary evaluation of this enzymatic method showed it to be precise at Hcy levels close to or higher than the values in dogs with renal or cardiac disorders that had the highest Hcy levels. By contrast, at low Hcy levels, which were recorded basically in control dogs, the method suffers from high imprecision. The s le of choice is serum. The use of icteric s les should be avoided, while hemoglobin and lipids have only a minor effect on Hcy measurement. In conclusion, the enzymatic method employed in the current study provides useful information in dogs and could be used to monitor cardiac and renal disorders, in which Hcy concentrations are elevated.
Publisher: Wiley
Date: 28-08-2011
Publisher: Elsevier BV
Date: 10-2013
Publisher: Wiley
Date: 29-10-2010
Publisher: American Veterinary Medical Association (AVMA)
Date: 10-2016
Abstract: OBJECTIVE To assess sensitivity and specificity of manual and automated measurements of reticulocyte percentage, number, and production index for classification of anemia in dogs. DESIGN Retrospective case series SAMPLE 174 blood smears from client-owned dogs with anemia collected between 1993 and 2013 for which reticulocyte parameters were determined manually (nonregenerative anemia, 22 preregenerative anemia, 23 regenerative anemia, 28) or with an automated laser-based counter (nonregenerative anemia, 66 preregenerative anemia, 17 regenerative anemia, 18). PROCEDURES Diagnostic performance was evaluated with receiver operating characteristic (ROC) curves by considering preregenerative anemia as nonregenerative or regenerative. Sensitivity, specificity, and positive likelihood ratio were calculated by use of cutoffs determined from ROC curves or published reference limits. RESULTS Considering preregenerative anemia as non regenerative, areas under the curve (AUCs) for reticulocyte percentage, number, and production index were 97%, 93%, and 91% for manual counting and 93%, 90%, and 93% for automated counting. Sensitivity, specificity, and positive likelihood ratio were 82% to 86%, 82% to 87%, and 4.6 to 6.4, respectively. Considering preregenerative anemia as regenerative, AUCs were 77%, 82%, and 80% for manual counting and 81%, 82%, and 92% for automated counting. Sensitivity, specificity, and positive likelihood ratio were 72% to 74%, 76 to 87%, and 2.7 to 6.2, respectively. CONCLUSIONS AND CLINICAL RELEVANCE Whereas all reticulocyte parameters identified regeneration in anemic dogs, the performance of specific parameters was dependent on the method used. Findings suggested that lower cutoffs than published reference limits are preferred for reticulocyte number and production index and higher cutoffs are preferred for reticulocyte percentage. Reticulocyte production index may be useful when the pretest probability of regeneration is moderate.
Publisher: Elsevier BV
Date: 12-2010
DOI: 10.1016/J.TVJL.2009.08.019
Abstract: Leishmania infantum interferes with the oxidative metabolism of phagocytes. In order to assess whether derivatives of reactive oxygen metabolites (d-ROMs) decrease due to infection or increase due to inflammation, d-ROMs were measured in serum collected from control dogs (Group 1 n = 12), from dogs seropositive for Leishmania either symptomatic (Group 2 n = 27) or not (Group 3 n = 14), and from dogs with other diseases (Group 4 n = 16). The concentrations of d-ROMs in the four groups, expressed in Carratelli Units (U CARR) were, respectively, 75.4 ± 39.5 (median, 81.6), 108.2 ± 96.3 (73.4), 73.5 ± 62.2 (62.0), 127.7 ± 97.3 (94.3). There were no significant differences between groups, but dogs with values higher than the reference interval were found, mostly in Groups 2 and 4 (which had serum C-reactive protein levels consistent with inflammation), whilst low values were occasionally found in Groups 2 and 3. Inflammation may mask decreases in d-ROMs induced by Leishmania infection.
Publisher: Elsevier BV
Date: 2014
DOI: 10.1016/J.TVJL.2013.10.007
Abstract: This study was designed to determine if the activity of paraoxonase (PON1), an antioxidant enzyme that works as a negative acute phase reactant, is a better predictor for the clinical recovery of leishmaniotic dogs receiving standard treatments compared with inflammatory markers such as C reactive protein (CRP) and electrophoretic fractions. For this purpose we tested 20 healthy dogs (controls) and 39 leishmaniotic dogs classified as sick (group A, n=23) or severely sick (group B, n=16) and tested at admission and after 3, 7, 14, 21, 28, 35 and 42 days. At admission, CRP and electrophoresis were altered in both groups, while PON1 activity was abnormal only in group B. There were no differences related to the outcome (mortality, complications or time of recovery). PON1 activity normalized in about 2 weeks in dogs that had abnormal values at admission and a final positive outcome CRP normalized in 4-6 weeks and electrophoretic fractions were still altered after 6 weeks. The results show that, at admission, inflammatory markers did not predict the outcome of leishmaniasis. PON1 activity decreased only in some dogs with systemic inflammation but not in those with mild leishmaniasis: when decreased, PON1 normalized earlier than other markers in dogs that responded to treatment. This finding most likely depends on the rapid decrease in oxidative phenomena. PON1 activity should therefore be tested on admission: if low values are recorded, severe inflammation may be suspected and PON1 measurement may be repeated during treatment to early identify responsive dogs.
Publisher: Wiley
Date: 28-07-2021
DOI: 10.1111/VCP.12994
Abstract: To date, little information is available about the effect of preanalytical factors on the urinary protein‐to‐creatinine (UPC) ratio in cats. We aimed to evaluate the effect of a commercially available cat litter, creatinine measurements at three different dilutions of urine, and different storage conditions on the UPC ratio in cats. Feline urine specimens were prospectively collected. Twenty‐two whole‐urine specimens were placed uncovered and in contact with cat litter for 1 hour 25 urine supernatants were diluted 1:10, 1:20, and 1:100 for creatinine measurements. The correlation, difference, agreement, and concordance in classifying specimens according to International Renal Interest Society staging were determined. Storage effects on UPC ratios were assessed in specimens stored for 6 hours at +20℃ (n = 20), 1 week at +4℃ (n = 20), and 3 months at −20℃ (n = 25). Specimens were also subjected to four freeze‐thaw cycles (n = 20). Results were compared, and clinical significance was assessed by comparing each UPC ratio to the inter‐assay range of the baseline value. Exposure to cat litter did not affect UPC ratios. A positive proportional bias was found in the 1:100 dilution compared with the 1:20 dilution however, concordance was high for all comparisons. At +20, +4℃, and after four repeated freeze‐thaw cycles, UPC ratios were stable. Compared with baseline values, UPC ratios decreased ( P .01) after 8 and 12 weeks at −20℃. However, all UPC ratios were within the inter‐assay variability of the baseline value. Exposure to cat litter did not affect UPC ratios, but further studies are necessary to evaluate other potential variables. The effects of the dilutions and storage conditions were clinically acceptable, although the 1:20 and 1:100 dilutions were not perfectly comparable.
Publisher: Elsevier BV
Date: 04-2015
DOI: 10.1016/J.TVJL.2015.01.029
Abstract: Quantification of proteinuria is a fundamental step in staging dogs with chronic kidney disease and in monitoring the course of disease or the efficacy of anti-proteinuric treatments. Analytical precision and accuracy of the proteinuria assessment could be affected by several factors such as biological variability, different operators and quality control materials. The aim of this study was to assess whether inter-laboratory variability could affect the urinary protein to creatinine (UPC) ratio and whether this variability may affect patient classification according to the International Renal Interest Society (IRIS) sub-staging system. The same urine s les were analysed in three different laboratories using different instruments and different reagent brands. The results of the three laboratories were highly correlated to each other although urinary protein (UP), urinary creatinine (UC) and the UPC ratio of one laboratory were found to be significantly higher than those of the other two. No significant differences between the other two laboratories were recorded. The concordance in classifying dogs according to the IRIS guidelines was good if all three proteinuria categories were analysed separately or if borderline proteinuric (BP) dogs were included in the proteinuric group, and very good if BP dogs were merged into the non-proteinuric group. The inter-laboratory variability in UPC ratio measurement was not so great as to impede the identification of proteinuric dogs, but may influence the estimation of the magnitude of proteinuria.
Publisher: Japanese Society of Veterinary Science
Date: 2017
DOI: 10.1292/JVMS.17-0023
Publisher: Wiley
Date: 14-08-2013
DOI: 10.1111/VCP.12073
Abstract: Serum activity of paraoxonase (PON1) decreases during inflammation in many species. Little information is available on paraoxon-based tests and the possible role of PON1 in dogs. The objectives of the study were to validate an automated paraoxon-based assay to measure PON1 activity in canine serum, to determine its stability under different storage conditions, to determine a reference interval (RI) in healthy dogs, and to assess whether PON1 is of comparable diagnostic value as C-reactive protein (CRP) and α2-globulins. Intra-assay and inter-assay imprecision, linearity under dilution (LUD), interference, and storage artifacts were evaluated. A PON1 RI was determined for healthy dogs, and PON1 activity, sensitivity, and specificity were compared with CRP and α2-globulins. Intra- and inter-assay CVs were below 1.6% and 7.8%, respectively. The LUD test fitted the linear model. PON1 activity measurements were increased after addition of hemolysates and lipids, and after storage for 12 hours at room temperature, 72 hours at 4°C, and 6 months at -20°C. PON1 activity and CRP or α2-globulins did not correlate well. PON1 activity decreased significantly only in dogs with very high CRP concentrations. In contrast to CRP and α2-globulins, PON1 activity was not significantly different between dogs with and without inflammation. The automated paraoxon-based method to assess serum canine PON1 activity was accurate and precise, but it was influenced by hemolysis, lipemia, and standard storage conditions. In this study, contrarily to CRP and α2-globulins, PON1 activity did not provide diagnostic value as a negative acute phase protein in dogs.
Publisher: Wiley
Date: 11-2010
Publisher: Wiley
Date: 15-05-2023
DOI: 10.1111/EVJ.13944
Publisher: Elsevier BV
Date: 03-2021
Publisher: Wiley
Date: 03-2008
Publisher: Elsevier BV
Date: 10-2014
Publisher: Elsevier BV
Date: 08-2013
DOI: 10.1016/J.TVJL.2013.01.034
Abstract: Early detection of inflammation in neonatal calves allows early intervention, which may reduce mortality. Paraoxonase-1 (PON1) is a negative acute phase protein in humans. The aims of this study were to investigate age-related variability in serum PON1 activity and its clinical usefulness in neonatal calves. In healthy calves (n=9), PON1 activity increased with age from 2 to 21 days of age. There was no significant increase in PON1 activity in healthy calves from days 21 to 120 (n=15), but PON1 activity was significantly higher in adult cattle (n=45). In sick calves, serum PON1 was significantly lower in calves 28- to 120-days-old with respiratory disease (n=8) in comparison with age matched controls (n=20 and n=15, respectively). These results support the role of PON1 as a negative acute phase protein in cattle.
Publisher: Elsevier BV
Date: 07-2018
Publisher: Wiley
Date: 02-2023
DOI: 10.1111/VCP.13238
Abstract: While clinical studies on acute phase proteins (APPs) have significantly increased in the last decade, and most commercial labs are now offering major APPs in their biochemical profiles, APP testing has not been widely adopted by veterinary clinical pathologists and veterinarians. Measurement of APP concentration is a useful marker for detecting the presence or absence of inflammation in cats with various diseases. APPs can also be reliably measured in different biological fluids (eg, effusions and urine) to improve their diagnostic utility. Measurement of APPs can be extremely beneficial in cats with feline infectious peritonitis (FIP) to discriminate between FIP and non‐FIP cats with similar clinical presentations. Additional benefits come from multiple and sequential measurements of APPs, particularly in the assessment of therapeutic efficacy. APPs are more sensitive than WBC counts for early detection of inflammation and to demonstrate an early remission or recurrence of the diseases. Given the potential utility of APPs, more studies are warranted, with a particular focus on the applications of APPs to guide the length of antimicrobial therapies, as suggested by the antimicrobial stewardship policy. New inflammatory markers have been discovered in human medicine, with a higher specificity for distinguishing between septic versus nonseptic inflammatory diseases. It is desirable that these new markers be investigated in veterinary medicine, to further test the power of APPs in diagnostic setting.
Publisher: Wiley
Date: 02-02-2015
DOI: 10.1111/VCP.12222
Publisher: SAGE Publications
Date: 22-03-2011
Abstract: In humans, the glycosylation pattern of serum and of membrane glycoproteins is associated with invasiveness of tumors: specifically, α2,6-sialylation and α2,3-sialylation are associated with metastasizing and nonmetastasizing tumors, respectively. In turn, the type of sialylation depends on the activity of α2,6 or α2,3 sialyltransferase (ST) enzymes. Because of the high prevalence of metastasizing tumors with biological behavior similar to the human counterpart, female dogs with metastasizing neoplasms could provide a good animal model for investigating the potential roles of sialic acid (Sia) and ST enzymes in the pathogenesis of metastatic tumors. The aims of this study were (1) to validate a solid-phase method based on lectin staining of serum and tissue homogenates to investigate sialylation and ST activity and (2) to compare the results obtained with this method and with lectin staining and to collect preliminary information on sialylation and ST activity in dogs with ( n = 8) and without ( n = 8) mammary tumors. The data recorded in healthy dogs revealed that serum and tissue glycoproteins are prevalently characterized by a α2,6 sialylation, but ST-α2,3 seems to be the most active enzyme in both s les. Sia-α2,3 and ST-α2,3 activity decreases in serum and tissues of dogs with tumors, especially in a dog with metastasis, suggesting that the equilibrium between ST-α2,6 and ST-α2,3 activity shifts toward the former, as reported in humans.
Publisher: Wiley
Date: 26-08-2016
DOI: 10.1111/VCP.12388
Abstract: Urinary protein-to-creatinine (UPC) ratio is an early diagnostic and prognostic marker of renal disease in dogs. Pyrogallol red molybdate (PRM) and Coomassie brilliant blue (CBB) are the most popular dye-binding assays for measurement of proteinuria. Published guidelines recommend strict cut-off points to substage patients with chronic renal diseases, irrespective of the assay applied. However, analytic variability and method-dependent differences could affect substaging of patients. The aims of this study were to analytically validate the CBB assay to evaluate possible method-dependent differences with PRM in urinary protein (UP) determination, and to assess the influence of such differences in substaging according to the International Renal Interest Society (IRIS). Urine was collected from healthy and proteinuric dogs. Intra-assay and inter-assay repeatability (imprecision), linearity under dilution (LUD), and spiking recovery (inaccuracy) were determined for the CBB assay. Split s les were measured with PRM and CBB, and agreement between methods and concordance in classification according to IRIS guidelines was determined. The CBB assay was precise (< 10%) at all urine protein concentrations after excluding outliers from the intra-assay precision assay of high urine protein concentrations. Acceptable accuracy was demonstrated with both LUD and spiking recovery test. Both UP and UPC determined by CBB were significantly higher (P < .0001) than those obtained with PRM, and both a constant and proportional bias were present. Concordance of IRIS substaging was only moderate. The CBB is precise and accurate, but the higher UPC obtained with CBB vs PRM may affect interpretation of the IRIS guidelines.
Publisher: Elsevier BV
Date: 07-2022
Publisher: Wiley
Date: 26-02-2016
DOI: 10.1111/VCP.12340
Abstract: A 13-year-old female Domestic Shorthair cat was presented to the Veterinary Teaching Hospital of the University of Milan for an interscapular mass suspected to be a mesenchymal malignant tumor. A preoperative CBC performed with Sysmex XT-2000iV showed leukocytosis with neutrophilia and eosinophilia. The Sysmex WBC/DIFF scattergram showed an additional, well-separated cluster of events between the neutrophil, eosinophil, and lymphocyte clusters. Blood smear evaluation revealed the presence of a significant number of basophils thus, it was hypothesized that the additional cluster could represent the basophilic population. A second CBC, 24 days later, showed the same pattern on the WBC/DIFF scattergram in the absence of leukocytosis and neutrophilia. After surgical excision of the mass, a definitive diagnosis of feline injection site sarcoma was made. To the author's knowledge, there are no previous reports about the identification of feline basophils in the WBC/DIFF scattergram of Sysmex XT-2000iV.
Publisher: Elsevier BV
Date: 10-2014
DOI: 10.1016/J.RVSC.2014.07.010
Abstract: Paraoxonase 1 (PON1) is a negative acute phase protein bound to high density lipoproteins (HDL) and during the acute phase response (APR) protects HDL from peroxidation. The aim of this study was to assess the relationship between PON1 and HDL in canine babesiosis, a disease characterized by oxidative damages and by an APR. PON1, HDL and C-reactive protein (CRP), were measured in blood collected from 15 controls and 29 dogs with babesiosis s led at admission, and on days 1 and 7 after treatment. At admission, PON1 and HDL were significantly lower in affected dogs. HDL concentration increased at day 1 while PON1 increased and CRP decreased at day 7. This suggests that the decrease of PON1 at admission is in part due to an increased consumption, the decreased HDL may depend on lipid peroxidation and its rapid increase after treatment may depend on the antioxidant activity of PON1.
Publisher: Elsevier BV
Date: 06-2021
Publisher: Elsevier BV
Date: 02-2015
DOI: 10.1016/J.RVSC.2014.11.011
Abstract: In order to assess whether the concentration of high density lipoproteins (HDLs) changes in leishmaniotic dogs before and after treatment, HDL cholesterol (HDL-Chol and HDL%), C reactive protein (CRP) and activity of the antioxidant enzyme paraoxonase (PON-1) were measured in sera from 10 controls and 10 leishmaniotic dogs. Seven of these latter were s led also 3, 7, 14, 21 and 28 days after treatment with antimonials and allopurinol. HDL-chol, and PON-1 were low in leishmaniotic dogs at admission and increased after treatment. HDL-chol and HDL% correlated positively with PON-1 and negatively with CRP suggesting that HDLs decrease through an oxidative mechanism. Therefore, HDLs may be used to monitor the magnitude of oxidation associated with inflammation in leishmaniotic dogs.
Publisher: Wiley
Date: 07-2009
Publisher: Wiley
Date: 11-2009
Publisher: Wiley
Date: 04-2013
DOI: 10.1136/VR.101171
Publisher: Springer Science and Business Media LLC
Date: 18-12-2019
DOI: 10.1038/S41598-019-55009-W
Abstract: Hepcidins are an evolutionarily conserved class of liver-expressed peptide, from which the twenty-five amino acid hormone, hepcidin-25 (herein hepcidin), has gained significant notoriety as the master regulator of iron homeostasis in mammals. Hepcidin maintains iron homeostasis by controlling the dietary absorption of iron and the mechanisms of recycling cellular iron stores. With the physiological significance of this hormone well established, it has emerged as an informative biomarker. In a comparison of the genome, transcriptome and peptidome of Canis lupis familiaris , we reveal the size of the hepcidin peptide in the canine, previous reports of which were contradictory to the evolutionary conservation predicted by genome annotation. Here, measurement of the peptide by mass spectrometry, following isolation from greyhound blood serum, revealed an amino acid sequence and peptide mass, differing from all accounts to date, yet demonstrating perfect sequence identity to that of the greater Canidae lineage of the Carnivora. Importantly, in the greyhound, the measured hepcidin peptide showed a similar temporal pattern to total serum iron, consistent with our understanding of hepcidin regulating iron homeostasis, in agreement with human diagnostics, and providing added translational evidence of the measured peptide being the iron regulatory hormone of the Canidae.
Publisher: Baghdad University College of Veterinary Medicine
Date: 28-06-2023
Abstract: Lymphoma is a cancer arising from B or T lymphocytes that are central immune system components. It is one of the three most common cancers encountered in the canine lymphoma affects middle-aged to older dogs and usually stems from lymphatic tissues, such as lymph nodes, lymphoid tissue, or spleen. Despite the advance in the management of canine lymphoma, a better understanding of the subtype and tumor aggressiveness is still crucial for improved clinical diagnosis to differentiate malignancy from hyperplastic conditions and to improve decision-making around treating and what treatment type to use. This study aimed to evaluate a potential novel biomarker related to iron metabolism, embryonic haemoglobin (HBE), for early diagnosis. Archived s les in combination with prospective s les collected from dogs with and without lymphoma were used in this study for the retrospective analyses of this tumor based on the same biomarker lified by real-time quantitative polymerase chain reaction. The HBE mRNA was aberrantly expressed in canine B and T cell lymphoma compared to the normal lymph node tissue and hyperplastic lymph nodes. In conclusion, this study identified a novel potential biomarker for improving lymphoma diagnosis and treatment in dogs. Further studies with larger s le sizes are needed to confirm the suitability of this biomarker for canine lymphoma diagnosis.
Publisher: SAGE Publications
Date: 17-08-2020
Abstract: Paraoxonase 1 (PON1) is an inflammation marker associated with lipid oxidation and is used as a diagnostic marker in people. There is no information about the suitable substrate and analytic performance in cats, or its biological behavior compared with other inflammation markers. Our aims were to validate a paraoxon-based method to measure PON1 activity in feline serum, to assess stability of PON1 under different storage conditions and the impact of interfering elements, to determine a reference interval (RI) for healthy cats, and to correlate PON1 activity with 2 major acute-phase proteins. Intra- and inter-assay precision, accuracy, and RI were assessed using fresh serum. The same specimens were stored at room temperature, refrigerated, or frozen, and retested at defined intervals. Hemolysis, lipemia, and icterus were simulated to study interferences. PON1 results were compared to serum amyloid A (SAA) and alpha-1-acid glycoprotein (AGP) results. Analytical validation yielded precise and accurate results. PON1 activity is stable for up to 24 h at room temperature and up to 48 h at 4°C. Freezing at −20°C results in an increase after 72 h, with return to baseline values after 1 wk, that again increases after 6 mo. Only hyperlipemia interfered with PON1 activity. The RI based on 71 healthy cats was 58–154 U/L. PON1 activity was negatively correlated with AGP, but not with SAA. Serum PON1 activity can be measured accurately in cats, and it acts as a negative acute-phase protein.
Publisher: Elsevier BV
Date: 10-2013
DOI: 10.1016/J.TVJL.2013.06.022
Abstract: This study was aimed at determining the serum concentration of homocysteine (Hcy) and big endothelin-1 (big ET-1, the precursor of endothelin) in dogs with chronic kidney disease (CKD) with and without hypertension, proteinuria and inflammation, in order to explore their role as biomarkers of hypertension associated with CKD. Hcy and big ET-1 were measured using an enzyme-linked immunosorbent assay and an enzymatic cyclic reaction, respectively, in dogs with CKD staged, as proposed by the International Renal Interest Society (IRIS), using serum creatinine, urinary protein to creatinine (UPC) ratio and systolic blood pressure, and classified as affected or not by inflammation based on the serum concentration of C-reactive protein (CRP). Serum Hcy was significantly higher in dogs of IRIS stages II, III and IV compared with controls and in proteinuric compared with non-proteinuric dogs. No differences relating to the degree of hypertension or to the CRP concentration were found. Serum big ET-1 significantly increased in dogs of IRIS stage IV compared with controls, in proteinuric compared with non-proteinuric dogs, in dogs with severe hypertension compared with those without hypertension, and in dogs with increased CRP compared to those with normal CRP concentrations. Hcy only correlated with serum creatinine but big ET-1 significantly correlated with serum creatinine, UPC ratio, systolic blood pressure, and increased CRP. In conclusion, both Hcy and big ET-1 increase in dogs with CKD. Although further research is needed, big ET-1, but not Hcy, may also be considered as a biomarker of hypertension.
Publisher: Frontiers Media SA
Date: 08-08-2022
DOI: 10.3389/FVETS.2022.875339
Abstract: To compare concentrations of biomarkers of allergy [mast cell tryptase (MCT) and histamine], inflammation [interleukin (IL)-6,-10, and−18, CXCL8, CCL2, keratinocyte chemoattractant (KC), C-reactive protein (CRP)], endothelial glycocalyx shedding (hyaluronan), coagulation [prothrombin time, activated partial thromboplastin time, fibrinogen concentration, and von Willebrand Factor antigen, protein C (PC) and antithrombin (AT) activity], and hepatopathy [alanine transaminase (ALT), aspartate transaminase (AST), alkaline phosphatase (ALP), and total bilirubin] between dogs with anaphylaxis after suspected insect exposure, dogs with critical illness, and healthy dogs. This was a single center prospective clinical observational comparative biomarker study that included 25 dogs with anaphylaxis (evidence of insect exposure, acute dermatological signs, and other organ involvement), 30 dogs with other critical illness, and 20 healthy dogs. Differences across groups in biomarker concentrations were tested using one-way ANOVA or Kruskal-Wallis test, with significant P values (& .05) reported for pairwise differences detected by post-hoc tests. Logistic regression models were used to calculate the area under the receiver operator characteristic curve (AUROC) for discrimination between anaphylaxis and non-anaphylactic illness. Histamine concentration was significantly higher in the anaphylaxis group than the healthy ( P & 0.001) and critically ill groups ( P & 0.001), whereas no differences in MCT were detected amongst groups. Biomarker concentrations that were increased relative to healthy dogs in both the anaphylaxis and critically ill groups included IL-10 ( P & 0.001 and P = 0.007, respectively), CCL2 ( P = 0.007 and P & 0.001, respectively) and AST (both P & 0.001), whereas only the critically ill group had significantly increased CRP ( P & 0.001), IL-6 ( P & 0.001), KC ( P & 0.001), ALP ( P & 0.001), and fibrinogen ( P = 0.016) concentrations, compared to the healthy group. Only dogs with anaphylaxis had significantly higher hyaluronan ( P = 0.021) and ALT ( P = 0.021) concentrations, and lower PC ( P = 0.030) and AT ( P = 0.032) activities, compared to healthy dogs. Both CRP and histamine concentration showed good discrimination between anaphylaxis and other critical illness, with an AUROC of 0.96 (95% CI 0.91–1) and 0.81 (95% CI 0.69–0.93), respectively. This preliminary study in dogs with anaphylaxis after suspected insect exposure, found evidence of an early innate immune response, glycocalyx shedding and anticoagulant consumption. Both CRP and histamine showed potential clinical utility for differentiation between anaphylaxis and other critical illness.
Publisher: Elsevier BV
Date: 10-2021
Publisher: American Veterinary Medical Association (AVMA)
Date: 06-2012
Abstract: Objective —To determine whether preanalytic and analytic factors affect evaluation of the urinary protein-to-creatinine (UPC) ratio in dogs. S le —50 canine urine s les. Procedures —The UPC ratio was measured to assess the intra-assay imprecision (20 measurements within a single session), the influence of predilution (1:10, 1:20, and 1:100) for urine creatinine concentration measurement, and the effect of storage at room temperature (approx 20°C), 4°C, and −20°C. Results —The coefficient of variation at room temperature determined with the 1:20 predilution was 10.0%, with the highest coefficients of variation found in s les with a low protein concentration or low urine specific gravity. This variability could result in misclassification of s les with UPC ratios close to the thresholds defined by the International Renal Interest Society to classify dogs as nonproteinuric (0.2), borderline proteinuric (0.21 to 0.50), or proteinuric ( 0.51). A proportional bias was found in s les prediluted 1:10, compared with s les prediluted 1:20 or 1:100. At room temperature, the UPC ratio did not significantly increase after 2 and 4 hours. After 12 hours at room temperature and at 4°C, the UPC ratio significantly increased. The UPC ratio did not significantly change during 3 months of storage at −20°C. Conclusions and Clinical Relevance —The intra-assay precision of the UPC ratio was sufficiently low to avoid misclassification of s les, except for values close to 0.2 or 0.5. The optimal predilution ratio for urine creatinine concentration measurement was 1:20. A 1:100 predilution is recommended in s les with a urine specific gravity 1.030. The UPC ratio must be measured as soon as s les are collected. Alternatively, s les should be immediately frozen to increase their stability and minimize the risk of misclassification of proteinuria.
Publisher: Elsevier BV
Date: 06-2011
Publisher: SAGE Publications
Date: 05-07-2020
Abstract: Interest is growing in measurement of novel biomarkers for the diagnosis of acute kidney injury. Multiplex assays may provide a rapid and cost-effective way of measurement however, sparse information is published regarding their use in dogs. We aimed to validate a commercial magnetic bead–based assay for 5 biomarkers: clusterin (Clus), cystatin C (CysC), kidney injury molecule 1 (KIM-1), monocyte chemoattractant protein 1 (MCP-1), and neutrophil gelatinase–associated lipocalin (NGAL). Intra- and inter-assay imprecision, linearity under dilution (LUD), spike recovery (S-R), and hemoglobin interference were evaluated using serum from healthy and diseased dogs. Additionally, the effect of s le type (serum vs. plasma) was investigated. All values for Clus and MCP-1 were outside the assay’s measurable range. Intra- and inter-assay precision were acceptable for NGAL (CVs 8.8% and 13.2%, respectively). Regression analysis of LUD and S-R indicated good linearity for CysC and NGAL. Hemolysis did not affect measurement of any biomarker. Measured concentrations of CysC ( p = 0.018) and NGAL ( p = 0.015) were significantly lower in sodium citrate plasma compared to serum. We conclude that this magnetic bead–based assay is precise and accurate for NGAL measurement in canine serum. Inappropriate standards for MCP-1 and Clus, and poor accuracy for KIM-1 measurement, suggest that this assay cannot reliably quantify those biomarkers in canine blood. Measurements of CysC in canine blood using this assay must be interpreted with caution given inter-assay imprecision.
Publisher: Wiley
Date: 12-2007
No related grants have been discovered for Gabriele Rossi.