Asta Tvarijonaviciute

Acute phase protein response in goats

Acute phase proteins (APPs) are important diagnostic indicators of inflammatory disturbances in animals. The objectives of the current study were to validate analytical methods for measuring haptoglobin (Hp), serum amyloid A (SAA), acid soluble glycoprotein (ASG), fibrinogen, and albumin concentrations in goats and to evaluate their response to an inflammatory stimulus in this species. Intra- and interassay coefficients of variation (CVs) were in the range 0.07–9.31% and 1.83–12.68%, respectively, for all APPs and showed good precision. All assays determined APPs in a linear manner at different sample dilutions with high correlation coefficients with the exception of fibrinogen, which was measured by the heat precipitation method. Subcutaneous injection of turpentine oil induced an increase in Hp, SAA, ASG, and fibrinogen serum concentrations and a decrease in albumin concentration.

Obesity-related metabolic dysfunction in dogs: a comparison with human metabolic syndrome

BackgroundRecently, metabolic syndrome (MS) has gained attention in human metabolic medicine given its associations with development of type 2 diabetes mellitus and cardiovascular disease. Canine obesity is associated with the development of insulin resistance, dyslipidaemia, and mild hypertension, but the authors are not aware of any existing studies examining the existence or prevalence of MS in obese dogs.Thirty-five obese dogs were assessed before and after weight loss (median percentage loss 29%, range 10-44%). The diagnostic criteria of the International Diabetes Federation were modified in order to define canine obesity-related metabolic dysfunction (ORMD), which included a measure of adiposity (using a 9-point body condition score [BCS]), systolic blood pressure, fasting plasma cholesterol, plasma triglyceride, and fasting plasma glucose. By way of comparison, total body fat mass was measured by dual-energy X-ray absorptiometry, whilst total adiponectin, fasting insulin, and high-sensitivity C-reactive protein (hsCRP) were measured using validated assays.ResultsSystolic blood pressure (P = 0.008), cholesterol (P = 0.003), triglyceride (P = 0.018), and fasting insulin (P < 0.001) all decreased after weight loss, whilst plasma total adiponectin increased (P = 0.001). However, hsCRP did not change with weight loss. Prior to weight loss, 7 dogs were defined as having ORMD, and there was no difference in total fat mass between these dogs and those who did not meet the criteria for ORMD. However, plasma adiponectin concentration was less (P = 0.031), and plasma insulin concentration was greater (P = 0.030) in ORMD dogs.ConclusionsIn this study, approximately 20% of obese dogs suffer from ORMD, and this is characterized by hypoadiponectinaemia and hyperinsulinaemia. These studies can form the basis of further investigations to determine path genetic mechanisms and the health significance for dogs, in terms of disease associations and outcomes of weight loss.

Serum paraoxonase 1 (PON1) measurement: an update

Paraoxonase 1 (PON1) is a widely studied enzyme based on its protective role against poisoning by organophosphate (OP) metabolites of specific OP insecticides and in vascular disease, as well as its use as biomarker of diseases involving oxidative stress, inflammation and liver diseases.This review provides an update about the current knowledge in the field of the analytical procedures that are used for PON1 measurements. It will be specially focused on: (a) characteristics of the different substrates used for measuring PON1, with emphasis in four aspects: toxicity, polymorphism influence, rate of hydrolysis and diagnostic performance. And (b) the technical aspects of PON1 assays, in which the reagents and reaction conditions, sources of variation, quality control systems, equipment and interferences with other esterases will be discussed.The information provided in this review can contribute to a more accurate and safe measurements of PON1 in laboratories and encourage researchers to explore the wide areas of PON1 in veterinary medicine that are still unknown.

Validation of spectrophotometric assays for serum paraoxonase type-1 measurement in dogs.

OBJECTIVE To evaluate and validate 3 spectrophotometric assays for measuring serum activity of paraoxonase type-1 (PON1), an enzyme associated with high-density lipoproteins, in dogs. ANIMALS 22 healthy adult dogs and 10 dogs with eccentrocytosis. PROCEDURES 2 methods were adapted for use in 96-well microplates with phenyl acetate and 5-thiobutyl butyrolactonase as substrates, and 1 was adapted for use in an automated analyzer with p-nitrophenyl acetate as substrate. Blood samples were collected from all dogs, serum was harvested, and serum PON1 activity was measured with each method. RESULTS Imprecision was low for all 3 methods, with the exception of interassay imprecision for 5-thiobutyl butyrolactonase, and results were linear across serial sample dilutions. The 3 methods were able to detect low PON1 activity when EDTA was used for blood sample collection, yielded lower PON1 values in sick dogs with eccentrocytosis than in healthy dogs, and yielded highly correlated results. CONCLUSIONS AND CLINICAL RELEVANCE The methods described here may allow a wider use of PON1 activity as a biomarker of oxidative stress in dogs in clinical and research settings. Results of each method were robust and precise (with the exception of the interassay values for the lactonase method), and the methods were easy to set up in a laboratory.

Effects of weight loss in obese cats on biochemical analytes related to inflammation and glucose homeostasis.

The aim of the current study was to measure circulating metabolic and inflammation-related biochemical analytes in obese cats before and after weight loss. Thirty-seven overweight neutered cats were studied, median body weight 6.85 kg (range, 4.70 to 10.30 kg), representing a range of ages and both sexes. An individualized weight-loss program was devised for each cat and monitored until completion. Body fat mass was determined by dual-energy x-ray absorptiometry, whereas plasma concentrations of acute-phase proteins (APPs; eg, haptoglobin and serum amyloid A), hormones (eg, insulin, IGF-1, and adiponectin), and enzymes (eg, butyrylcholinesterase and paraoxonase type 1 [PON-1]) associated with inflammation and metabolic compounds (eg, glucose) were also measured. No significant changes were found in APPs after weight loss (P > 0.3), but significant increases in plasma adiponectin (P = 0.021) and IGF-1 (P = 0.036) were seen, whereas insulin (P < 0.001) and homeostasis model assessment (P = 0.005) decreased significantly. Plasma concentrations before weight loss of PON-1 (P = 0.004), adiponectin (P = 0.02), and IGF-1 (P = 0.048) were less in cats that failed to complete weight loss than cats that were successful, whereas glucose concentration was greater. Finally, multivariable linear regression analysis showed that lean tissue loss during weight management was associated with percentage weight loss (greater weight loss, greater lean tissue loss; R = 0.71, P < 0.001) and plasma adiponectin concentration before weight loss (lesser adiponectin, more lean tissue loss; R = -0.52, P = 0.023). In conclusion, various metabolic abnormalities occur in feline obesity, and these can be linked to outcomes of weight-loss programs. The changes that occur with weight loss suggest an improved metabolic status.

Serum acute phase proteins as clinical phase indicators and outcome predictors in naturally occurring canine monocytic ehrlichiosis.

BACKGROUND Canine monocytic ehrlichiosis (CME), caused by Ehrlichia canis, is an important tick-borne disease of global importance. Currently, limited information is available on the diagnostic and prognostic value of acute phase proteins (APPs) in dogs naturally infected with E. canis. HYPOTHESIS APPs may be useful indicators of the clinical phase of CME and predictive of the clinical outcome (death or survival). ANIMALS Fifty-six dogs naturally infected with E. canis and 7 clinically healthy control dogs. METHODS C-reactive protein (CRP), serum amyloid A (SAA), haptoglobin (Hp), and albumin concentrations determined on admission were retrospectively compared among 27 dogs with nonmyelosuppressive CME, 29 dogs with myelosuppressive CME and 7 healthy dogs. Diagnosis of CME was based on clinical and clinicopathological findings, seropositivity to E. canis, polymerase chain reaction amplification of E. canis-specific 16S rDNA, microscopic observation of Ehrlichia sp. morulae in blood monocytes or some combination of these. RESULTS Mean concentrations of CRP, SAA, and Hp were significantly higher in the myelosuppressed dogs compared with the other groups, but no significant differences were found in the concentration of albumin. Survival analysis of the affected animals indicated that APP concentrations were not associated with clinical outcome; the latter was strongly associated with pancytopenia (odds ratio for death 22.7) and neutropenia (odds ratio for death 7.7). CONCLUSIONS AND CLINICAL IMPORTANCE CRP, SAA, and Hp serum concentrations on admission are useful indicators of the clinical phase of CME, but are not useful predictors of clinical outcome.

Prognostic value of serum acute‐phase proteins in dogs with parvoviral enteritis

OBJECTIVE To evaluate the acute-phase protein response in dogs with parvoviral enteritis as predictor of the clinical outcome. METHODS Canine parvovirus infection was diagnosed based on the compatible clinical findings and confirmed by the canine parvovirus antigen test in 43 dogs of less than six months of age. Blood samples for complete blood cell count and acute-phase proteins (C-reactive protein, haptoglobin, ceruloplasmin and albumin) were collected before treatment. Twenty-three dogs died during or after treatment (non-survival) and the rest recovered (survival). Five healthy dogs were enrolled as control. RESULTS Serum C-reactive protein, ceruloplasmin and haptoglobin levels in dogs with parvoviral enteritis were higher (P<0·001, P<0·01 and P<0·001, respectively), but serum albumin was lower (P<0·001) than those in controls. Mean C-reactive protein and ceruloplasmin values in non-survival were higher (P<0·01) than those for survival dogs. C-reactive protein was found to be superior to ceruloplasmin, haptoglobin and albumin for distinguishing survival from non-survival dogs. Values higher than 92·4 mg/l for C-reactive protein had a sensitivity of 91% to predict mortality. CLINICAL SIGNIFICANCE The magnitude of the increase in serum acute-phase proteins in dogs with parvoviral enteritis could be a useful indicator of the prognosis of the disease. In acute-phase proteins, C-reactive protein is a potent predictor of mortality in dogs with parvoviral enteritis.

Effect of Weight Loss in Obese Dogs on Indicators of Renal Function or Disease

BACKGROUND Obesity is a common medical disorder in dogs, and can predispose to a number of diseases. Human obesity is a risk factor for the development and progression of chronic kidney disease. OBJECTIVES To investigate the possible association of weight loss on plasma and renal biomarkers of kidney health. ANIMALS Thirty-seven obese dogs that lost weight were included in the study. METHODS Prospective observational study. Three novel biomarkers of renal functional impairment, disease, or both (homocysteine, cystatin C, and clusterin), in addition to traditional markers of chronic renal failure (serum urea and creatinine, urine specific gravity [USG], urine protein-creatinine ratio [UPCR], and urine albumin corrected by creatinine [UAC]) before and after weight loss in dogs with naturally occurring obesity were investigated. RESULTS Urea (P = .043) and USG (P = .012) were both greater after weight loss than before loss, whilst UPCR, UAC, and creatinine were less after weight loss (P = .032, P = .006, and P = .026, respectively). Homocysteine (P < .001), cystatin C (P < .001) and clusterin (P < .001) all decreased upon weight loss. Multiple linear regression analysis revealed associations between percentage weight loss (greater weight loss, more lean tissue loss; r = -0.67, r(2) = 0.45, P < .001) and before-loss plasma clusterin concentration (greater clusterin, more lean tissue loss; r = 0.48, r(2) = 0.23, P = .003). CONCLUSION AND CLINICAL IMPORTANCE These results suggest possible subclinical alterations in renal function in canine obesity, which improve with weight loss. Further work is required to determine the nature of these alterations and, most notably, the reason for the association between before loss plasma clusterin and subsequent lean tissue loss during weight management.

Serum acute phase protein concentrations in female dogs with mammary tumors.

Acute phase proteins (APPs) are proteins whose concentrations in serum change after any inflammatory stimulus or tissue damage. The aim of the current study was to evaluate 3 positive APPs (C-reactive protein, serum amyloid A, and haptoglobin) and 1 negative APP (albumin) in female dogs with mammary neoplasia. Acute phase proteins were studied in 70 female dogs aged 8–12 years in the following groups: healthy (n = 10); mammary tumors in stages I (n = 19), II (n = 5), III (n = 6), IV (n = 5), and V (n = 7); and with mammary neoplasia plus a concomitant disease (n = 18). In animals with mammary neoplasia, significant increases of positive APPs were only detected in those that had metastasis or a neoplasm with a diameter greater than 5 cm and ulceration. Dogs with mammary neoplasia and a concomitant disease also had high C-reactive protein concentrations. Albumin concentration was decreased in animals with metastasis and with a concomitant disease. The results of the present study indicate that the acute phase response could be stimulated in female dogs with mammary gland tumors because of different factors, such as metastasis, large size of the primary mass, and ulceration or secondary inflammation of the neoplasm.

Effect of weight loss on inflammatory biomarkers in obese dogs

The objective of this study was to evaluate the effects of weight loss on selected serum inflammatory biomarkers in obese dogs. An experimentally induced bodyweight reduction of approximately 2.5%/week was accompanied by significant decreases in metabolic markers of obesity (lipidic profile, fructosamine, and insulin-like growth factor-1). The concentrations of acute phase proteins and of selected cytokines remained within reference ranges in obese dogs during weight loss, suggesting that significant inflammation was not a major component of this experimental model. However, adiponectin concentrations increased following the period of weight loss suggesting reduced susceptibility of these animals to obesity-related inflammation.