Pharmacological Treatment of Perianal Gland Tumors in Male Dogs.

The presence of androgen (AR) and estrogen (ER) receptors has been demonstrated both in normal perianal (hepatoid) glands and in perianal tumors. The aim of this study was to demonstrate the relationship between the expression of AR and ER in perianal gland tumors and the effectiveness of antihormonal treatment. The study was performed on 41 male dogs with neoplastic lesions of the anal region. Histopathological evaluation of the lesions revealed 24 adenomas, 12 epitheliomas, and five carcinomas. Treatment was administered orally with tamoxifen at a dose of 1 mg/kg BW and cyproterone acetate at a dose of 5 mg/kg. Tumor diameters were measured regularly with calipers and recorded in millimeters starting with the measurement before treatment, and then after 1, 2, 3, 6, 12, 18, and 24 months of therapy. The results show that hepatoid adenomas that are characterized by high expression of AR and ER receptors respond positively to antihormonal therapy, resulting in complete tumor regression. For locally malignant hepatoid epitheliomas and carcinomas with low expression of AR and ER receptors, antihormonal therapy makes it possible to reduce the size of the tumor, but does not make it possible to cure it completely.

Comparative evaluation of cytokine and acute-phase protein concentrations in sera of dairy cows with subclinical and clinical ketosis as a different view of the causes of the disease.

BACKGROUND AND AIM: Ketosis is a common disease occurring during the first stage of lactation in highly productive dairy cows. The aim of the present study was the comparative assessment of selected pro-inflammatory cytokines (including tumor necrosis factor-α [TNF-α] and interleukin 6 [IL-6]), anti-inflammatory cytokines (including IL-10), and acute-phase proteins (APPs) (including haptoglobin [Hp] and serum amyloid A [SAA]), in the sera of cows with subclinical ketosis (SCK), in those with clinical ketosis (CK), and in healthy cows. MATERIALS AND METHODS: Thirty dairy cows of Holstein-Friesian breed were investigated. The cows were divided into three groups depending on the serum ß-hydroxybutyric acid (ßHBA) level. The control, SCK, and CK groups included healthy cows, cows with SCK, and cows with CK, respectively. BHBA concentration in blood serum was determined using colorimetric method. The blood serum was used for proper tests. Cytokine (TNF-α, IL-6, and IL-10) and APPs (SAA and Hp) concentrations in the investigated samples were determined by enzyme-linked immunosorbent assay method. RESULTS: The SCK group had significantly higher TNF-α, IL-6; IL-10, and SAA values than had the CK group (p<0.01). The SCK group had a lower Hp concentration than had the CK group (p<0.05). CONCLUSION: This study showed that the inflammation intensity is higher in the initial phase of the disease and decreases during the advancement, probably due to active anti-inflammatory mechanisms (an increase of IL-10 concentration), which protect animal organism from self-destruction. On the basis of our study, it can be assumed that ketosis development in dairy cows was preceded by the systemic inflammation that may influence the progress of this disease.

Androgen and Estrogen Receptor Expression in Different Types of Perianal Gland Tumors in Male Dogs.

Perianal gland tumors are modified sebaceous glands present in the skin of the perianal region in the dog. Hormonal stimulation may induce hyperplasia of the perianal glands or their neoplastic progression. The presence of androgen (AR) and estrogen (ER) receptors have been demonstrated both in normal perianal glands as well as in perianal tumors. The aim of the study was an immunohistochemical assessment of the expression of estrogen and androgen receptors in perianal gland tumors in dogs as an applicatory marker for antihormonal treatment. Biopsy samples of perianal masses were collected from 41 male dogs. A histopathological examination revealed 24 adenomas, 12 epitheliomas and five carcinomas. The immunohistochemical staining showed a mainly nuclear expression of AR and ER in the neoplastic cells. Both the androgen and estrogen receptors were expressed in adenoma, epithelioma and carcinoma cases; however, the highest expression of the receptors was stated in the adenoma and epithelioma. In the case of the carcinoma, the expression of sex hormone receptors was very weak. The differences of the number of cells expressing AR and ER as well as the observed differentiated intensity of staining in the studies demonstrated that the determination of the expression of the sex hormone receptors may be useful to elaborate a diagnostic and therapeutic algorithm.

Selected leukocyte subpopulations in peripheral blood and uterine washings in cows before and after intrauterine administration of cefapirin and methisoprinol.

The aim of the study was to evaluate the selected lymphocyte subpopulations TCD4, TCD8, BCD21, BCD25, CD18, CD11b, and MHC II in blood and uterine flush of cows with endometritis, before and after intrauterine (i.u.) administration of cefapirin and methisoprinol. The research was carried out on 28 cows with clinical endometritis. Animals were divided into four groups, each composed of seven cows, depending on the i.u. preparation used: Group A, cefapirin; Group B, methisoprinol; Group C, cefapirin and methisoprinol simultaneously; and a control group-without medication. The study was performed using flow cytometry method. Summarizing the results of the research, i.u. infusion of cefapirin caused a weakening of the effector phase of the local uterine immune response; however, it enhanced leukocyte chemotaxis and antigen presentation. After i.u. administration of methisoprinol, the stimulation of specific uterine immunity mechanisms was mainly observed. The use of both mentioned preparations showed the strengthening of specific uterine immunological mechanisms presumably caused by methisoprinol, despite the inhibitory effect of the antibiotic. Intrauterine use of immunostimulatory substances can improve the effectiveness of the endometritis treatment in cows by improving specific local mechanisms of uterine immunity. As a consequence, it may enhance the effector function of immune competent cells and finally eliminate inflammation.

DNA Adduct Assessment During Antihormonal Treatment of Perianal Gland Tumors With Tamoxifen in Male Dogs.

BACKGROUND/AIM: Determination of DNA adduct count was performed in mononuclear cells during antihormonal treatment of perianal gland tumors. MATERIALS AND METHODS: Eight- to fifteen-year-old dogs with carcinoma (CAR Group; N=5), epithelioma (EPI Group; N=16) or adenoma (ADE Group; N=24) were used. The control group suffered from perineal hernia or rectal diverticulum (CTR Group; N=25). Blood was collected at baseline, and at one and six months after the beginning of the anti-hormonal treatment with tamoxifen (1 mg/kg of body weight). DNA adduct count was determined using autoradiography. RESULTS: At baseline, DNA adduct count reached the highest value in the CTR Group, and the lowest in the EPI Group (p<0.05). Six-month-long therapy with tamoxifen resulted in a significant increase in the DNA adduct count by 78.7%, 221.5% and 198.3% in the ADE, EPI and CAR groups, respectively (p<0.05). CONCLUSION: Increased DNA adduct formation after long-term administration of tamoxifen shows its genotoxicity.

Phagocytic and oxidative burst activity of phagocytic cells in peripheral blood and uterine washings in cows with clinical endometritis before and after intrauterine use of cephapirin and methisoprinol.

The aim of the study was to evaluate phagocytic and killing activity of phagocytic cells in blood and uterine flush of cows with endometritis before and after intrauterine (i.u.) administration of cephapirin and methisoprinol. The research was carried out on 28 cows with clinical endometritis. Animals were divided into four groups, each composed of seven cows, depending on the i.u. treatment used: Group A-cephapirin; Group B-methisoprinol; Group C-cephapirin and methisoprinol at the same time; and a control group-without medication. Using flow cytometry technique, the phagocytic activity of granulocytes and monocytes was identified, as well as the oxidative burst activity of neutrophils in the peripheral blood and uterine washings. Summarizing the results of the research, i.u. infusion of cephapirin caused a reduction in the phagocytic and killing activity of phagocytes. The i.u. use of methisoprinol increased phagocytic and killing activity of phagocytes in the uterus. Administering both listed substances simultaneously showed a decrease in phagocytosis, presumably due to the dominating inhibitor effect of the antibiotic. However, also an increase of mean fluorescence intensity was observed, presumably caused by the methisoprinol. Intrauterine use of immunostimulatory substances, can improve the effectiveness of the treatment of endometritis in cows.

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.

Morphological, densitometric and mechanical properties of pelvic limb bones in 14-month-old female ostriches (Struthio camelus).

Limited information on physiological characteristic of bones in ostrich skeleton are available, even though bone weaknesses in ostriches are commonplace. Thus, the aim of this study was to evaluate morphological, densitometric, and mechanical properties of pelvic limb long bones (femur, tibia, and tarsometatarsus) in 14-month-old female ostriches (Struthio camelus var. domesticus). After the slaughter procedure, all bones were isolated, cleaned of soft tissues, and bone weight and length were determined. Using dual energy X-ray absorptiometry (DEXA) bone mineral density (BMD) and bone mineral content (BMC) were determined. Volumetric bone mineral density (vBMD) and geometrical parameters of the bones were determined using the quantitative computed tomography (QCT) method. Maximum elastic strength and ultimate strength of the bones were determined using three-point bending test. Significant differences were revealed between morphological, densitometric, and mechanical properties of femur, tibia, and tarsometatarsus in female ostriches at the slaughter age of 14 months. Elaborated experimental model and determination of morphological, densitometric, and mechanical properties of femur, tibia, and tarsometatarsus in female ostriches may serve for further studies on metabolic regulation of skeletal system properties with environmental, physiological, dietary, pharmacological, and toxicological factors.

Diagnostic and prognostic value of cellular proliferation assessment with Ki-67 protein in dogs suffering from benign and malignant perianal tumors.

In the perianal region of carnivores, skin consists of modified sebaceous glands called perianal glands. Tumors originating from perianal glands are the third most frequent type of neoplasm in male dogs after neoplastic diseases of testes and skin. Ki-67 is a nuclear non-histone protein considered a proliferation marker in normal and neoplastic proliferating cells. Previous investigations revealed that Ki-67 expression may be used as a prognostic factor for breast cancer in humans. Thus, the aim of this study was to estimate the diagnostic and prognostic value of Ki-67 evaluation in dogs suffering from benign and malignant perianal tumors. The highest value of the Ki-67 index was obtained in the carcinoma group (18.50% ± 2.68), significantly higher compared to the values obtained in the control tissue (7.63% ± 2.12) and adenoma (7.33% ± 1.06; all P < 0.05). Statistically significant differences in the Ki-67 index were not found between the epithelioma group (11.95% ± 1.96) and all other groups (P < 0.05). This investigation on dogs with perianal gland tumors has shown significantly increased expression of Ki-67 antigen in carcinoma cells, while the expression of this protein was similar in the case of control tissues, adenoma and epithelioma. Thus, it may be postulated that Ki-67 evaluation in perianal gland tumors in dogs may serve as a useful marker possessing high diagnostic and prognostic value and enabling differentiation of malignant and benign tumors.

VEGF and 17-ß-estradiol levels after tamoxifen administration in canine hepatoid gland adenomas and hepatoid gland epitheliomas.

The aim of the present study was to determine the serum levels of vascular endothelial growth factor in animals in the course of pharmacological treatment against perianal gland neoplasms. Research material comprised of tumor tissue samples obtained from 30 dogs and blood drawn from dogs with tumors and control group animals. The neoplasm type was determined in accordance with the relevant WHO classification. Immunoenzimatic determination of VEGF levels in the blood sera was performed. In all studied animals suffering from tumors, pharmacological tamoxifen treatment was administered, at a dosage of 2 mg/kg bodyweight. The medication was administered for one month. In order to monitor the serum levels of 17-ß-estradiol and VEGF, blood was drawn from sick animals three times (on the day of the diagnosis, as well as at one and six months after treatment). The VEGF determination assay was performed in accordance with the manufacturer's guidelines for the ELISA. In the studied group, 12 animals were diagnosed with hepatoid gland adenomas and 18 with hepatoid gland epitheliomas. Elevated VEGF levels were observed in the group of dogs with hepatoid gland ephithelioma in comparison with the control group. In the studied groups, a decrease in serum VEGF level and a complete remission of neoplastic lesions was observed one month after administering tamoxifen. The VEGF levels in dogs with hepatoid gland adenoma continued to decline with time. In the case of dogs with hepatoid gland epithelioma, after the initial drop one month after treatment, a rapid increase of the growth factor level was observed, which was significantly higher in animals suffering a relapse of the neoplastic disease (50% of dogs). A significant correlation was observed between 17-ß-estradiol and VEGF levels in dogs with hepatoid gland epithelioma on the day of diagnosis (Rxy=0.64, p<0.05) and six months after treatment (Rxy=0.54, p<0.05). Conclusion: VEGF overexpresion observed six months after tamoxifen treatment may constitute a prognostic factor in terms of the progression of the neoplastic process. The level of VEGF correlates with the level of 17-ß-estradiol in serum. Apart from anti-estrogen effects, tamoxifen also demonstrates anti-angiogenic activity.

Serum concentration of magnesium in dogs suffering from tumors of the perianal glands.

The aim of this study was to determine serum magnesium concentrations in dogs with benign and malignant tumors of the perianal glands. The magnesium concentration was determined in samples taken from 45 tumor-affected and 17 healthy males. Twenty one dogs suffered from malignant tumors (Malignant group), while benign changes were diagnosed in 24 cases (Non-malignant group). All dogs with neoplastic disease were treated with antiandrogens and antiestrogens. Determination of magnesium concentrations in serum obtained from dogs before the start of treatment (baseline), and one and six months later was performed using atomic absorption spectrophotometry. One-way ANOVA and the post-hoc Tukey's HSD test were used to compare differences between the groups investigated and time-related changes. Significantly higher serum magnesium concentrations were found in the group with malignant tumors when compared to the control and non-malignant groups (P<0.001). Serum magnesium concentrations in dogs with non-malignant neoplastic changes had increased gradually by 17% and 41% at one and six months when compared to the baseline values (P<0.05). In conclusion, the malignant neoplastic process in dogs was associated with a higher baseline serum magnesium concentration. Thus, determination of the serum magnesium concentration might be helpful for diagnostic differentiation between malignant and benign perianal tumors in dogs.

Ornithine alpha-ketoglutarate increases mineralization and mechanical properties of tibia in turkeys.

Skeletal disorders in rapidly growing poultry are commonplace. This study was performed to investigate the effect of ornithine alpha-ketoglutarate (OKG) administration during the last 7 weeks of life on structural properties, mineralization, and mechanical endurance of skeleton in turkeys at slaughter. Healthy HB Medium Bronze female turkeys were randomly assigned to two weight-matched groups at the age of 12 weeks. OKG was administered orally to the experimental group (N=17) at the dose of 0.4 g/kg body weight per day, while the control group (N=16) received an equal dose of the vehicle. The turkeys were slaughtered at the age of 19 weeks and the tibiae were isolated for analysis. The effect of OKG on skeletal system development in turkeys was evaluated in relation to both geometrical and mechanical properties as well as quantitative computed tomography (QCT). Free amino acids concentrations were assessed with the use of ion-exchange chromatography. Significantly increased bone mineral density of the trabecular and the cortical bone of tibia in the turkeys given OKG for the last 7 weeks of production cycle were observed (P<0.05). OKG administration improved mechanical endurance of the tibia estimated by the three-point bending test (P<0.01). Plasma amino acid analyses showed increased level of aspartate, proline, alanine, valine, isoleucine, leucine, and ornithine (all P<0.05) after OKG treatment, whereas cystathionine concentration was decreased (P=0.03). Obtained results indicate that oral OKG administration has beneficial effects on skeletal development in fast growing turkeys and this effect is connected with increased amino acid synthesis. These observations may serve to improve skeletal properties in birds, especially when considering that skeletal disorders often affect the tibia and the proper function of the skeletal system plays an essential role in animal welfare and poultry production.

Magnesium concentrations in mammary tumours in dogs.

The neoplasmic process causes the disturbance of magnesium metabolism in the organism. The studies were conducted in order to determine magnesium concentration in the tissues and serum of dogs with malignant and non-malignant mammary tumours in comparison to its concentration in the tissue and serum of healthy dogs. From the studies presented in the paper it turns out that magnesium concentration in the mammary neoplasmic tissue increases with the increase of tumour malignancy and in comparison to its concentration in the control tissue. Magnesium concentration in the blood serum decreases with the increase of tumour malignancy and in comparison to the control tissue. Therefore it was shown that there is a connection between the degree of tumour malignancy and magnesium concentration in the tissue and serum.