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We aimed to assess the usefulness of monitoring inter-alpha trypsin inhibitor heavy chain 4 (ITIH4) and haptoglobin (Hp) in peripheral and local blood in canine pyometra, and evaluation the relationships among acute phase proteins (APPs), systemic inflammatory response syndrome (SIRS) and the presence of bacteria. The material was collected from bitches with pyometra and from healthy ones. Blood was taken from the cephalic and uterine veins. APPs levels were quantified by ELISA. In the peripheral circulation, the Hp was higher in animals with open-cervix pyometra (OCP) than in the closed-cervix pyometra (CCP) and the control group. The Hp concentration was not correlated with age, with the presence of SIRS or with the type of bacteria (Gram-negative, Gram-positive or mixed flora). The ITIH4 concentrations in the peripheral blood did not differ significantly in the cases of pyometra. The Hp concentration in the local circulation increased in the OCP but not in the CCP groups, although the histopathological changes in the endometrium were similar. Peripheral Hp concentrations may be a useful tool in differentiating between the types of pyometra.

The main aim of this report was to investigate and compare the response of serum C-reactive protein (CRP) and ferritin, two positive acute phase proteins (APPs) which usually show an increase in inflammatory processes, in dogs with pyometra. For this purpose, two different studies were made. In the first one , both proteins were measured together in an APPs profile in 25 dogs with pyometra, 25 dogs with pancreatitis (as an example of a positive inflammatory control group), and in 25 healthy dogs. In the second study, to advance the knowledge of the changes and evolution of serum ferritin and CRP in dogs with pyometra after treatment, the concentrations of both APPs were analyzed in 30 dogs with pyometra at diagnosis and after ovariohysterectomy and in 10 clinically healthy female dogs before and after elective spaying. In both studies, bitches with pyometra showed significant increases in serum CRP, indicating an inflammatory condition, but not in serum ferritin despite being a moderate positive APP. This divergence between the dynamics of these APPs could be a useful tool for the suspicion of cases of canine pyometra., (© 2023. The Author(s).)

This study aimed to compare the activity of selected glycosidases (β-galactosidase, α-l-fucosidase, β-N-acetyl-hexosaminidase, and sialidase) in homogenates of uterine tissues obtained from female dogs with and without pyometra. In addition, it examined the availability of substrates for these glycosidases in the homogenates. The study was carried out on female dogs undergoing ovariohysterectomy for pyometra (n = 10) and clinically healthy dogs (n = 10) undergoing elective spaying. The activity of β-galactosidase, α-l-fucosidase, and β-N-acetyl-hexosaminidase was analyzed using a spectrofluorometer and that of sialidase using a colorimetric method. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis with Alcian Blue (AB) and Periodic Acid-Schiff (PAS) staining was performed to determine the presence of substrates for these glycosidases in the homogenates of uterine tissues. The results revealed that the activity of all the examined glycosidases was significantly higher (P < 0.05) in the uterine tissues isolated from dogs with pyometra in comparison to healthy dogs. The electrophoretic patterns of the selected samples showed several proteins, which contained different sugar moieties stained by AB and PAS and the profiles differed significantly between the pyometra group and the healthy group. Densitometric analysis of AB staining showed patterns between 233 and 148, 86 and 55, and 43 and 20 kDa, which differed markedly in sugar content between the examined groups of animals. Similarly, PAS staining analysis revealed patterns of different molecular weights, between 233 and 117 and between 55 and 32 kDa, which also differed in sugar content. These findings suggest that canine pyometra is accompanied by the increase in the activity of selected glycosidases in the uterus. This could potentially modify the glycan structures of uterine glycoproteins and in result their biological functions. Further studies are needed to elucidate the potential role of the increased activity of glycosidases in the pathogenesis of this disease., Competing Interests: Declaration of competing interest The authors declare that there is no conflict of interest regarding the publication of this article., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Measurement of acute phase proteins (APPs) as biomarkers in canine medicine is in increasing demand. In the present study, the development and validation of two ELISA methods for the quantification of canine inter-alpha-trypsin inhibitor heavy chain 4 (ITIH4) and haptoglobin (Hp) are shown. The adequate imprecision and accuracy and wide analytical range make the developed methods appropriate to quantify ITIH4 and Hp in serum samples. The inter- and intra-assay CVs were lower than 10 %, and the assays maintained linearity under dilution and showed analytical equivalence with the method of radial immunodiffusion. The measurement of CRP, Hp and ITIH4 in sera from bitches affected by pyometra allowed us to determine that ITIH4 behaves as a moderate APP in dogs. The group of bitches affected by pyometra showed very high levels of CRP (147 ± 91 mg/L), corresponding to a strong inflammatory process, which resulted in a moderate increase in the concentrations of Hp (7 times) and ITIH4 (3 times) compared to the control group., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Adiponectin is a 30 kDa protein hormone that has anti-atherogenic properties, being an insulin-sensitizing and anti-inflammatory molecule. Salivary adiponectin concentrations correlate positively with serum, thus, saliva was indicated as appropriate biofluid for its measurement in different clinical situations. However, inflammation was indicated as main confounding factor when evaluating the usefulness and the reliability of determination of salivary adiponectin. The aim of the present report was to evaluate the dynamics of salivary and serum adiponectin in systemic non-septic and septic inflammation using a dogs as a model. Forty bitches were enrolled. Seventeen dogs were healthy (group I, non-septic) and 23 bitches were diagnosed with pyometra (group II, septic). Ovariohysterectomy was performed for all animals. Saliva and blood samples were collected before (D0) and 3 (D3) and 10 (D10) days after ovariohysterectomy. At D0, Group I showed higher serum and salivary adiponectin than group II, although statistical significance was only detected in salivary adiponectin between the two groups at D0 (P = 0.001). In serum, adiponectin was higher on D0 than on D3 and tended to reach pre-surgery values on D10 in both groups. Salivary adiponectin showed similar behaviour to serum in Group I, while in group II salivary adiponectin concentrations were lowest on D0 and tended to increase on D3 and D10. The data obtained in present study describe for the first time the comparative behavior of salivary adiponectin in non-septic and septic inflammation. Salivary, and not serum, adiponectin seemed to mimic better the inflammatory and general health status., (Copyright © 2019 Elsevier B.V. All rights reserved.)

The main aim of this study was to investigate the effect of pyometra on glycosylation of proteins in the uterine tissues from female dogs, using western blotting with selected lectins (Sambucus nigra agglutinin - SNA and Maackia amurensis agglutinin - MAL II). In addition protein pattern of examined tissues was also evaluated. The study was performed on 10 female dogs undergoing ovariohysterectomy because of pyometra and 10 clinically healthy female dogs, undergoing elective spaying (ovariohysterectomy). Uterine tissue samples of 1 cm 2 were taken from the middle region of each uterine horn in both group of animals immediately after ovariohysterectomy. Tissue samples were homogenized and analysed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and western blotting with SNA and MAL II. SDS-PAGE analysis showed differences between pyometra samples and controls in the amount of obtained protein fractions and the protein content in the individual fractions. Five protein (with a molecular weight of 193.78 kDa, 103.18 kDa, 77.67 kDa, 70.39 kDa, and 53.00 kDa) were found only in the pyometra samples. The remaining fractions differed in intensity of staining, which indicated differ abundance of a given protein. The results of western blotting with SNA and MAL II demonstrated that the pattern obtained from densitometric analysis differs between adequate healthy and pyometra samples with regard to the amount of protein fraction obtained as well as the intensity of staining of particular fraction. The pyometra tissues contained seven SNA-binding proteins (with a molecular weight 189.94 kDa, 165.51 kDa, 100.94 kDa, 59.42 KDa, 41.32 kDa, 35.16 kDa, and 32.6 kDa) that were not in the healthy tissues. Of the nine remaining fractions, six showed significantly higher (P < 0.05) intensity of staining in the healthy uterine tissues. In turn, the MAL II-binding protein with a molecular weight 75.85 kDa, 51.12 kDa, and 49.98 kDa were found only in the pyometra samples. Of the 28 remaining fractions, ten demonstrated significantly higher (P < 0.05), and five fractions had significantly lower (P < 0.05) intensity of staining in the pyometra tissues. The results obtained indicate that proteins in uterine tissues from female dogs with pyometra are differently glycosylated compared to normal uterine tissues. These findings provide the basis for further studies of the possible role of glycosylation in the pathogenesis of canine pyometra., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Background: Pyometra is a serious bacterial infection of the uterus affecting female dogs and manifests as an accumulation of pus in the uterine lumen. The aim of the study was to assess changes in serum interleukin (IL)-6 and IL-10 concentrations in bitches with pyometra undergoing ovariohysterectomy., Findings: Blood samples were collected from healthy bitches (controls) and bitches with pyometra before surgery, and 3 and 10 days after ovariohysterectomy. Before surgery, bitches with pyometra had significantly higher serum concentrations of IL-6 and IL-10 than the controls. After surgery, the serum concentration of IL-6 and IL-10 decreased significantly. In healthy dogs, the concentration of IL-6 and IL-10 showed a significant increase 3 days after surgery followed by a decrease on day 10., Conclusion: An increase in serum concentrations of IL-6 and IL-10 was present before surgery in bitches with pyometra and 3 days after ovariohysterectomy in healthy controls. Concentrations decreased after ovariohysterectomy and/or proper healing, suggesting that these cytokines can be useful for assessment of the postoperative period in bitches.

This study compares serum concentrations of tryptophan (TRP), kynurenine (KYN), kynurenic acid (KYNA), and indoleamine 2,3-dioxygenase (IDO) activity in healthy bitches and bitches with bacterial uterine infection (pyometra). The effects of surgery were also assessed by measuring these variables in both groups of dogs before and after ovariohysterectomy. Presurgery, mean (±standard deviation) TRP, KYN, and KYNA concentrations and IDO activity were 68.44 ± 21.77, 2.00 ± 0.33, 112.11 ± 111.91 μmol/L, and 29.22 ± 10.10, respectively, in the healthy dogs; and 40.16 ± 12.11, 8.27 ± 3.94, 411.11 ± 199.60 μmol/L, and 205.92 ± 154.20, respectively, in the dogs with pyometra. Tryptophan and KYN levels had normalized on suture removal (10 days after surgery) though IDO activity and KYNA concentrations remained elevated during the postoperative period compared with presurgery values in both study groups. Our results suggest that KYNA concentrations and IDO activity could be useful indicators of the inflammation induced by pyometra and could be also used to monitor recovery following ovariohysterectomy in both healthy dogs and dogs with pyometra., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Pyometra is one of the most common diseases of the reproductive system in bitches. The inflammatory processes occurring in the uterus result in changes in concentrations of numerous serum biomarkers, which are used for monitoring the postoperative period. The aim of the present report was to study the evolution of serum levels of insulin-like growth factor-1 (IGF-1) and C-reactive protein (CRP) in bitches suffered from pyometra and after ovariohysterectomy in comparison with the control group of healthy dogs submitted to the surgery. Our results indicate that the serum level of IGF-1 is decreased (101.6 ng/mL [73.8-177.8 ng/mL]), whereas CRP is increased (114.6 μg/mL [95.3-131.6 μg/mL]) during severe inflammation. When inflammation caused by pyometra and surgery is resolved, the level of IGF-1 is increased (186.0 ng/mL [94.6-344.3 ng/mL], P < 0.05) and CRP decreased (23.3 μg/mL [9.9-77.0 μg/mL], P < 0.01), suggesting that these markers could be useful for assessment of the postoperative period in bitches., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Pyometra is a life-threatening reproductive disorder that affects the uterus of female dogs. This study was designed to identify the possible indicators of uterine inflammation by comparing C-reactive protein (CRP), serum amyloid A (SAA), and haptoglobin (Hp) concentrations in uterine arterial and peripheral venous blood in bitches with open- and closed-cervix pyometra. CRP, SAA, and Hp concentrations were higher in bitches with closed-cervix pyometra irrespective of the site of blood collection. Higher acute-phase protein concentrations were observed in peripheral compared with uterine arterial blood in bitches with closed-cervix pyometra, whereas the levels were comparable in dogs with open-cervix pyometra. Our results indicate that mean acute-phase protein concentrations differ according to pyometra type/severity and blood source and suggest the possible use of peripheral blood levels of CRP, SAA, and Hp to monitor inflammation during the course of pyometra., (Copyright © 2013 Elsevier Inc. All rights reserved.)

Kynurenic acid (KYNA) is produced enzymatically in humans and animals from kynurenine. Reports concerning changes of kynurenine metabolism during inflammation are available in the literature. Pyometra is a pathological condition characterized by the accumulation of pus in the uterine lumen and bacterial infection. The objective of the study was to compare the serum and endometrial KYNA concentrations in healthy bitches and those with pyometra. KYNA was determined by means of high-performance liquid chromatography with fluorometric detection. The serum content of KYNA in bitches with pyometra was significantly higher than in healthy bitches. The KYNA content in the endometrium of bitches with pyometra was higher, yet the difference was not statistically significant. Our result indicates that determination of KYNA might be a marker of pyometra in bitches.

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Abstract: The aim of the study was to assess changes in serum C-reactive protein (CRP), serum amyloid A component (SAA) and haptoglobin (Hp) concentrations in healthy bitches and in those with pyometra undergoing ovariohysterectomy, and to establish the usefulness of such determinations for monitoring the postoperative period. Our results indicate that CRP and SAA determinations serve to evaluate the severity of the inflammatory process in pyometra since the concentrations of these acute phase proteins were increased immediately after surgery and diminished thereafter. The CRP and SAA response was rapidly produced while Hp concentrations increased in a more gradual manner. Thus, postoperative concentrations of CRP and SAA provide valuable information on the subsidence of the inflammatory response during the uneventful postoperative period. Our findings also suggest that acute phase proteins might be useful diagnostic markers of early postoperative complications. [Copyright &y& Elsevier]