A Pilot Study of the Role of Selected Biomarkers of Kidney Injury in Dogs with Dilated Cardiomyopathy.

Heart and kidney diseases are among the most frequent medical conditions diagnosed in small animals. Due to the functional interconnection between these organs, the concept of the cardio-renal axis has been developed. In this context, renal disease or dysfunction often occurs secondary to heart diseases, such as dilated cardiomyopathy (DCM). DCM is the most common cardiomyopathy and a leading cause of mortality in large-breed dogs. Traditional biomarkers like creatinine or symmetric dimethylarginine concentration are not always effective, especially in the early stages of the disease, underscoring the need for more sensitive markers of renal impairment during heart failure (HF). This study aimed to evaluate the efficacy of selected biomarkers as indicators for early kidney damage in dogs with stage B2 DCM. We measured serum concentrations of cystatin C, KIM-1 (kidney injury molecule-1), and NGAL (neutrophil gelatinase-associated lipocalin) and their ratios to creatinine, analyzing their diagnostic values. Cystatin C was quantified using a sandwich enzyme immunoassay, while KIM-1 and NGAL were measured with enzyme-linked immunosorbent assay kits designed for canine diagnostics. The concentrations were indexed against serum creatinine. The study included 26 dogs: 9 with HF and 17 healthy controls. The mean ± standard deviation for healthy dogs for cystatin C, cystatin C/creatinine ratio, KIM-1, KIM-1/creatinine ratio, NGAL, and NGAL/creatinine ratio were 0.24 ± 0.04, 0.26 ± 0.07, 0.61 ± 0.07, 0.67 ± 0.13, 2.76 ± 1.8, and 2.79 ± 1.81, respectively. For DCM dogs, these values were 0.27 ± 0.1, 0.32 ± 0.12, 0.61 ± 0.08, 0.69 ± 0.17, 6.46 ± 5.22 (p = 0.02), and 7.99 ± 6.53 (p = 0.04). This study's findings suggest that during the asymptomatic phase of DCM, only NGAL concentration and the NGAL/creatinine ratio may serve as diagnostic markers for early-stage kidney injury.

The diagnostic value of selected immune parameters in peripheral blood of dogs with malignant mammary tumours - a preliminary study.

Introduction: The main adaptive immune cells are T and B lymphocytes and they play key roles in the induction of immune responses against canine mammary tumours. Investigating these cell subpopulations may lead to more precise diagnosis of these malignancies. Material and Methods: The percentages of CD3+, CD4+ and CD8+ T cells and of CD21+ B cells in the peripheral blood of bitches with malignant mammary tumours were compared with those in the blood of healthy animals. The phenotypic features of peripheral blood leukocytes were evaluated by flow cytometry. Results: There was a significant difference in the mean percentages of CD3+ lymphocytes between healthy (66.7%) and metastatic dogs (46.1%), and between tumour-bearing non-metastatic (66.6%) and metastatic dogs. There was also a significant difference in CD4+ T helper cell percentages between healthy dogs (40.4%) and dogs with metastases (23.2%), and between the latter and dogs without them (35.5%). In the case of CD21+ lymphocyte subsets, a significant difference was noted between healthy animals (10.9%) and those with metastases (20.1%), and between the latter and patients without metastases (8.5%). There were also significant differences in CD3+/CD21+ ratios between the group with metastases (3.0), the healthy group (7.8), and the group without metastases (8.5). Similarly, a significant difference was noted in CD4+/CD8+ ratios between animals with metastases (1.4), bitches in the control group (2.2), and dogs without metastases (1.9). Conclusion: Peripheral blood leukocyte phenotypic characteristics are putative novel biomarkers. These findings may be useful in future studies improving mammary tumour diagnostic procedures, especially in metastasis detection.

Variation of Physiological and Behavioural Parameters during the Oestrous Cycle in Mares.

The behaviour of oestrous mares is well-known in horse breeding. However, alterations in their physiological and behavioural indices during the whole oestrous cycle are scarcely known. The objective of the study was to analyse changes in cardiac activity variables, rectal and superficial temperatures, behaviour towards humans and conspecifics, and the time of standing and locomotor activity in mares during their oestrous cycle. Fifteen adult mares in oestrus were examined in the morning and evening (six successive days) and in dioestrus (five days-once every third day). The oscillation of physiological and behavioural variables accompanies changes in mares' sexual behaviour. Most physiological variables studied in oestrus indicate the elevated activity of the adrenergic nervous system and, opposite to that, both behaviour towards humans and conspecifics and the time of standing relate to a relaxed state. The end of oestrus, manifested by a rapid decrease in most of the physiological variables studied, is followed by changes of behavioural variables at the beginning of dioestrus. The time of locomotor activity arises at the end of oestrus. The outcomes may contribute to the knowledge of, among others, mare owners who evaluate the oestrus by mares' sexual behaviours without regarding other rhythmically changing variables.

Structure and Pathologies of Articular Cartilage.

The aim of the review was to describe a complex microstructure and biomechanical properties of the articular cartilage as well as a current review of its pathologies encountered in veterinary practice. The articular cartilage with its unique features: complex microarchitecture, significant mechanical durability and elasticity, lacking blood, lymphatic vessels, and innervation, seems to stand in contradiction to the laws of biology. It can be involved in a vast majority of diseases, from osteoarthrosis as a result of natural aging process to more complex in nature like osteochondromatosis. The primary role of articular cartilage is to provide the surface for movement in any single joint in the body. Therefore, its diseases lead to physical impairment and deterioration of the quality of life. Treatment of articular cartilage poses a formidable challenge in both modern human and animal medicine.

Effects of Topical Treatment of Foot Rot in Sheep Using Ozonated Olive Ointment.

INTRODUCTION: Foot rot in small ruminants is highly contagious, causes severe lameness, and impairs fertility and wool and meat production. It is usually treated with parenteral antibiotics, with attendant antibiotic resistance risk, and with bactericidal footbaths, potentially harmful to humans and the environment. An alternative treatment in sheep is proposed based on repeated topical ozonated ointment application. Its effectiveness and safety were evaluated by estimation of acute-phase response, biochemical indicators of organic damage, and antioxidant/oxidant balance (AOB). MATERIAL AND METHODS: The study was conducted on ten sheep with Egerton scale 2-3 lesions. Ozone application was repeated every day for seven days. Blood was drawn first (T0) after foot cleaning and before ozonation, then (T1) seven days after the first ozone application, and finally (T2) four days after the last application. RESULTS: High clinical effectiveness was observed, with total recovery by 28 days from the start of treatment. A significant increase in antiradical activity was noted on the basis of a 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) assay from 1.16 ± 0.04 µmolTe/mL at T0 to 1.23 ± 0.03 µmolTe/mL at T1, with a slight decrease in oxidative stress. Calculated on the basis of antiradical capacity, AOB was higher at T1 (130 ± 19%) and decreased to 110 ± 16% at T2. Calculated on the basis of reducing power, it was 169 ± 22% at T1 and 131 ± 17% at T2. CONCLUSION: These results indicated that the AOB is efficient enough to prevent oxidative organ injury and the applied doses of ozone are safe for animals.

The Use of Allogenic Stromal Vascular Fraction (SVF) Cells in Degenerative Joint Disease of the Spine in Dogs.

BACKGROUND/AIM: Stem cells are widely used in regenerative medicine and in clinical practice for the treatment of damaged nerve tissue, myocytes, tendons, and ligaments. The aim of the study was to monitor VEGF levels after the administration of allogenic cellular material (SVF) in the course of treatment of dogs suffering from degenerative joint disease in the spinal region. MATERIALS AND METHODS: The study was conducted on 10 dogs of both genders, aged between 6 and 13 years in which allogenic stromal vascular fraction of stem cells (SVF) was administered intravenously. The control group was composed of 10 clinically healthy dogs. Before treatment and after 2- and 8-week intervals blood samples were obtained from the study group dogs in order to determine VEGF levels via immunoenzymatic test. RESULTS: in a few days after the therapy, alleviation of pain symptoms and reduction of lameness were noticed. The VEGF level in 2 weeks after the therapy was significantly elevated (median: 38.77 pg/ml), while in 8 weeks a decrease was observed (median: 18.37 pg/ml). Conlusion: Administration of allogenic stem cells has a positive influence on elevation of the VEGF levels in the blood serum of affected animals as well as their regeneration capacity.

Elevated EGF Levels in the Blood Serum of Dogs with Periodontal Diseases and Oral Tumours.

BACKGROUND/AIM: Paradontopathy and neoplasms of the oral cavity represent one of the greatest challenges in human and animal dentistry. EGF plays a key role in maintaining the integrity and proper rate of cell proliferation in normal oral epithelium. The aim of the present study was to study serum levels of EGF in dogs diagnosed with periodontal diseases and oral cavity tumours. MATERIALS AND METHODS: The samples comprised of cancerous tissue sections and serum obtained from dogs of various breeds, aged between 5-13 years. Serum EGF concentrations were measured by an immunoenzymatic method. RESULTS: The median for EGF concentration in serum of dogs suffered from severe periodontal diseases was greater when compared to the control group. EGF concentration in dogs with malignant tumours was significantly higher than in those with non-malignant growths. A positive correlation between EGF concentration and tumour size was also observed. EGF level in dogs diagnosed with benign tumours was comparable to the control group. CONCLUSION: The blood serum level of EGF increases significantly in patients with malignant oral tumours and advanced periodontal disease. In malignant tumours, the high level of EGF correlates with the size and invasiveness of the neoplasm.

EGF Level in Hepatoid Gland Adenomas and Hepatoid Gland Epitheliomas in Dogs After Administering Tamoxifen.

BACKGROUND/AIM: Neoplastic lesions of perianal glands account for approximately 10% of all skin cancer cases in dogs. They occur in many dog breeds, usually in male animals aged over 6 years. Due to their hormone-dependency, tamoxifen can be used in antineoplastic treatment. The aim of the study was to measure epidermal growth factor (EGF) levels in the serum of dogs with perianal tumours after tamoxifen treatment and to use it as a prognostic factor for further treatment. MATERIALS AND METHODS: The study was performed on 19 male dogs aged between 6 and 14 years, diagnosed with neoplastic hyperplasia in the perianal region. The control group comprised 10 healthy dogs brought in for routine castration. The research material comprised blood drawn from the animals and tumour specimens for histopathology. The study group received 1-month treatment with tamoxifen. Blood serum was then tested for 17-ß oestradiol level, and for EGF level on the first day of the therapy and 6 months after treatment completion. RESULTS: Hepatoid gland adenomas were diagnosed in 10 cases, and hepatoid gland epitheliomas in nine cases. Elevated 17-ß oestradiol levels were observed in all dogs. On the first day of treatment with tamoxifen, the serum EGF levels in all study groups were higher than in the control group. At the 6-month follow-up, the EGF levels were significantly reduced in hepatoid gland adenoma cases compared to those taken on the first day of treatment of tamoxifen, while in animals with hepatoid gland epithelioma, it was greatly increased and was correlated with relapse. CONCLUSION: Perianal gland tumours are characterised by EGF overexpression, which can be helpful in early-stage prognosis and treatment. An increase in EGF levels 6 months after tamoxifen therapy correlates with disease progression and may be a useful prognostic factor.

Treatment of Articular Cartilage Defects: Focus on Tissue Engineering.

The treatment of articular cartilage defects seems to be one of the greatest challenges in modern orthopaedics. From a medical point of view there are 3 main goals to achieve for cartilage trauma treatment: restoration of the joints motion, pain relief and elimination/delay of the onset of osteoarthritis. Treatment can be divided into conservative (including pharmacotherapy, arthrocentesis and physiotherapy) and surgical. The last comprises reparative techniques, regenerative methods and symptomatic treatment. While both are focused on reconstruction of the damaged cartilage, the difference lies in the type of filling tissue. Reparative techniques include: drilling of the subchondral bone, spongiolisation, abrasion, mictrofracture, and autologous matrix induced chondrogenesis (AMIC). Regenerative methods contain: periosteal and perichondral grafts, mosaicplasty, autologous chondrocyte implantation and matrix-induced autologous chondrocyte implantation (MACI). Nowadays tissue engineering, including gene therapy, is emerging as one of the key approaches to cartilage treatment and holds promises for new achievements and better outcomes of many cartilage diseases and traumas.

Application of Platelet-rich Plasma and Tricalcium Phosphate in the Treatment of Comminuted Fractures in Animals.

AIM: To assess the applicability of ß-tri-calcium phosphate (TCP) and platelet-rich plasma (PRP) in the treatment of comminuted fractures in small animals. MATERIAL AND METHODS: The experimental study was carried out on 16 New Zealand White rabbits. After creating the bone defect and performing tibial osteotomy, TCP implants containing activated PRP were introduced into the fracture and the defect. The fracture was stabilised using external fixators or intramedullary nails. After 12 weeks, the animals were euthanised, and radiological, histological, scanning electron microscopy and peripheral quantitative computed tomography examinations were performed. The analysis also covered the results of fracture treatment in 37 small animals (cats and dogs) in which treatment with TCP containing PRP was used as an alternative to cancellous bone implantation. RESULTS: Correct bone union was observed in the experimental groups, TCP remained visible at the site of the fracture after 12 weeks. In the clinical application in small animals, bone union was observed in over 91% of treated animals. CONCLUSION: ß-TCP and activated PRP may be an effective method of bone union enhancement in the treatment of comminuted fractures in small animals.

Osteochondral Repair Using Porous Three-dimensional Nanocomposite Scaffolds in a Rabbit Model.

AIM: To evaluate the utility of a novel nanocomposite biomaterial consisting of poly-L/D-lactide, and hydroxyapatite bioceramics, enriched with sodium alginate in articular cartilage defect treatment. MATERIALS AND METHODS: The biomaterial was prepared using the method of solvent casting and particle leaching. The study was conducted on 20 New Zealand White rabbits. Experimental osteochondral defects were created in the femoral trochlear grooves and filled with biomaterials. In control groups, the defects were left to spontaneously heal. The quality of newly-formed tissue was evaluated on the basis of macroscopic and histological assessment. Additionally the level of osteogenic and cartilage degradation markers were measured. RESULTS: The majority of the defects from the treatment group were covered with tissue similar in structure and colour to healthy cartilage, whereas in the control group, tissue was uneven, and not integrated into the surrounding cartilage. CONCLUSION: The results obtained validate the choice of biomaterial used in this study as well as the method of its application.