Pathomorphological and immunohistochemical study of selected markers of tumour cell proliferation in equine sarcoids

The purpose of the study was a pathomorphological and immunohistochemical analysis of tumour cells and connective tissue in equine sarcoids. Investigations were performed using histopathological, ultrastructural, immunohistochemical (PCNA, p53, cytokeratin, vimentin) and histochemical (Ag-NORs) methods. The study was conducted on 50 sarcoids originating from 36 horses and classified as occult, verrucous, fibroblastic and a mixed type of sarcoid based on their clinical appearance. Most of the tumours were located on the girth (30%), neck (24%), head (12%), and legs (12%). The average age of the horses at the first clinical examination was 5.7 years. The sarcoids occurred on the skin of mares (61%), geldings (31%) and stallions (8%), the predominant was Wielkopolska breed (41%) and mixed breeds with Wielkopolska breed (41%). The predominant colour was bay (80%). The data showed that the presence of characteristic, microscopic features was variable but it was not consistent enough to allow differentiation of the clinical types based on histopathology. PCNA expression was not characteristic for the clinical type of sarcoid but it appeared to be a useful tool for the determination of the biological activity of the tumour and the probability of its recurrence. No relationship was found between AgNORs and cell proliferation. The study demonstrated the presence of p53 positive cells in the epidermal and fibroblastic portions. Numerous p53-positive cells were observed in the sarcoids and tended to recurrence. The staining for cytokeratin and vimentin makes the diagnosis of tumour easier. The immunohistochemical studies of PCNA, and p53 are of great significance to the prognosis.     

Contents of selected heavy metals in the liver, kidneys and abdominal muscle of the brown hare (Lepus europaeus Pallas, 1778) in Central Pomerania, Poland

The content of iron, zinc, copper, manganese, lead, and cadmium was determined in the kidney, liver, and abdominal muscle of the brown hare (Lepus europaeus), a species known for its bioindicative potential in environmental quality assessment. The animals assayed were divided into immature (in their first year of life; n=25) and adult (n=39). The hares were acquired in an area situated far away from major cities and direct impacts of industrial and transport pollution. The zinc content found in the assayed hare's muscles (25.9 microg/g w.w.) and that of manganese in the kidneys, liver, and muscles (2, 2.51 i 0.85 microg/g w.w., respectively) tended to be higher than those reported from the brown hare elsewhere in Europe. The content of cadmium (particularly in the kidneys) and lead proved substantial and close to those typical of the hare inhabiting industry-affected areas. Analysis of the data pooled for all the individuals showed Zn and Cd contents to increase with age in the kidneys, the liver Cd content increasing with age as well; on the other hand, the age-Cu content was negative. In addition, a number of significant correlations between the metals themselves were revealed, particularly with respect to Zn-Cd correlation in kidneys (r(s)=0.47), Fe-Cd, Zn-Cu, Mn-Zn, Mn-Cu, and Mn-Pb in the liver (r(s) of 0.42, 0.86, 0.72, 0.79, and 0.42, respectively), and Zn-Cu in the muscle (r(s)=0.56). The kidney cadmium content was higher by 81% in the adult than in the immature hare, the adult hare muscle copper content being lower by 15.5%.     

Electrolyte level and blood pH in dogs infected by various 18S RNA strains of Babaesia canis canis on the early stage of babesiosis

The purpose of the studies was to determine electrolyte disturbances and blood pH changes in dogs with babesiosis and possibly show a connection between the Babesia (B.) canis strain causing the infection and the intensity of these irregularities. 40 animals (group 1) with early babesiosis and 40 healthy dogs (group 2) were studied and their blood pH and blood levels of potassium, chlorides; calcium and sodium were determined. At the same time, molecular typing of parasites was carried out to detect which B.canis strain (18S RNA-A or 185 RNA-B) had caused the disease in dogs of group 1. In group 1, four dogs were acidaemic, twelve had normal blood pH, and 24 were alkalaemic. Potassium concentration was below normal in 16 out of 40 dogs (40%) and normal in 24 dogs. Hypochloremia was present in 36 out of 40 dogs; chloride was normal in the remaining four animals. Serum sodium concentration was low in 16 of 40 dogs, normal in 20 of 40 dogs and high in four dogs. Calcium concentration was normal in all 40 dogs. In dogs of group 2 no abnormalities of haematological or blood biochemical parameters were observed. 29 out of the 40 dogs of group 1 were infected with the 18S RNA-A strain and eleven with the 18S RNA-B strain of Babesia canis canis. We did not observe any correlation between the type of strain causing the infection and the electrolyte disturbances in the serum of sick dogs. Hypocalaemia was observed in ten specimen infected with 18S RNA-A and six infected with 18S RNA-B. Additionally, in dogs infected with 18S RNA-A, hypochloraemia (28), hyponatraemia (10), hypernatraemia (2) were observed, as well as blood pH drop (4) or increase (14). The 18S RNA-B-infected dogs suffered from hypochloraemia (8), hyponatraemia (6), hypernatraemia (2) and increase in blood pH (10).The studies conducted did not answer the question of whether the type of electrolyte disturbances in dogs with babesiosis can be connected with the strain of the parasite that induced the disease, as happens in the case of other clinical parameters (Adaszek et al., 2009). Further studies in this respect, conducted on a larger group of animals, are necessary.     

Apoptosis of peripheral blood leukocytes from rabbits infected with non-haemagglutinating strains of rabbit haemorrhagic disease virus (RHDV)

The report demonstrates that the induction of apoptosis in peripheral blood granulocytes and lymphocytes of rabbits infected with three non-haemagglutinating RHDV strains (English Rainham, German Frankfurt, and Spanish Asturias) is a crucial determinant of the pathogenesis of rabbit haemorrhagic disease. Apoptosis was measured by flow cytometric detection of caspase activity. These studies demonstrated that the investigated RHDV (rabbit haemorrhagic disease virus) viral strains affected leukocyte apoptosis to varying degrees. Enhanced leukocyte apoptosis was detected between 4 and 36h after infection and was more pronounced in lymphocytes than in granulocytes. The data presented here thus provide a preliminary understanding of the kinetics of apoptosis in leukocytes of rabbits infected with RHDV.     

Upregulation of LENK and VIP in paracervical ganglion neurons supplying the urinary bladder of tetrodotoxin- and resiniferatoxin-treated female pigs

Both resiniferatoxin (RTX) and tetrodotoxin (TTX) have been reported to be effective in several clinical trials aiming to cure urinary bladder dysfunction. The goal of this experiment was to study the effect of intravesical administration of RTX and TTX on the chemical coding of paracervical ganglion (PCG) neurons that supply the urinary bladder in pigs. The vasoactive intestinal peptide (VIP) and the opioid family member Leu5-enkephalin (LENK) are both known for their regulatory effects in the function of the porcine genitourinary tract. The PCG neurons innervating the urinary bladder were identified by application of the retrograde tracer Fast Blue (FB), injected into the bladder wall prior to intravesical RTX or TTX administration. Immunocytochemical detection of LENK and VIP expression in the FB-labelled perikarya revealed that in the control group 25.15% of the FB-positive PCG neurons contained LENK, and 9.22% of them expressed VIP. Intravesical infusion of RTX resulted in an increase in the number of LENKIR neurons to 48.19% and VIP-IR perikarya to 11.25%. Optional treatment with TTX induced increase of LENK-IR neurons up to 81.67% and VIP-IR population to 16.46% of the FB-positive PCG cells. The present results show that both neurotoxins affect the chemical coding of PCG nervous cells supplying the porcine urinary bladder and that they stimulate both LENK and VIP expression. Furthermore, the results indicate a possible involvement of LENK and VIP neurons in the mechanisms of action of RTX and TTX in the therapy of overactive bladder disorder.     

Genomic and genetic aspects of heart failure in dogs - a review

The most common causes of heart failure in dogs are valvular disease, predominantly endocardiosis, and myocardial disease, predominantly dilated cardiomyopathy. They are related to changes in the expression of several genes in the heart muscle and in peripheral blood nuclear cells which could be considered as prognostic or diagnostic markers of heart disease in dogs. Since many human genetic markers of heart failure have turned out to be useless in dogs, the screening for genomic markers of canine heart failure could give more insight into the molecular pathology of these diseases and aid the development of new treatment strategies.     

Effects of β-Hydroxy-β-Methylbutyrate and γ-Oryzanol on Blood Biochemical Markers in Exercising Thoroughbred Race Horses

In both the horse and the man, nutritional ergogenic aids have been used to improve physical ability in conjunction with an appropriate training regimen. Although training increases physical condition, the ease of taking a nutritional additive to improve training results explains the demand for supplementation, which may increase mechanical energy of work, delay onset of fatigue, or improve neuromuscular coordination. The purpose of this study was to determine the effects of oral supplementation of β-hydroxy-β-methylbutyrate (HMB) and γ-oryzanol (GO) on indices of exercise-induced muscle damage in Thoroughbred race horses. In this 32-week study, the horses were assigned to either a placebo, GO (3.0 g/d), HMB (15 g/d), or GO and HMB treatment groups. The supplements were administered for the first 16 weeks of the study during the training period before the racing season began. Blood samples were taken at baseline, and then during training, before exercise, immediately after exercise, and 30 minutes after exercise. Heart rate and speed were monitored in each exercise session. Hematocrit, glucose, lactate (LA), creatine phosphokinase, and aspartate aminotransferase were measured before and after each exercise session. Analysis of variance showed a significantly greater increase in postexercise creatine kinase activity in placebo-supplemented group than in the other treatment groups, both in the training period and during the racing seasons (P < .05). Blood LA was higher immediately after exercise in the placebo group compared with the supplemented groups. In conclusion, supplementation with HMB and GO resulted in decreased creatine kinase and LA after exercise. These findings support the hypothesis that HMB and GO supplementation helps to prevent exercise-induced muscle damage.     

Effects of the commercial extract of aronia on oxidative stress in blood platelets isolated from breast cancer patients after the surgery and various phases of the chemotherapy

Since the extract from berries of Aronia melanocarpa presents antioxidative properties in plasma and in blood platelets, not only from healthy group, but also from patients with benign breast diseases and in patients with invasive breast cancer before surgery, the aim of our present study was to evaluate the oxidative stress by measuring the level of various biomarkers of this process such as the generation of superoxide anion radicals (O(2)(-·)), the amount of carbonyl groups and 3-nitrotyrosine in proteins or the amount of glutathione in blood platelets isolated from breast cancer patients after the surgery and after various phases of the chemotherapy in the presence of A. melanocarpa extract (Aronox) in vitro. We demonstrated in platelet proteins from patients with invasive breast cancer (after the surgery and after various phases of the chemotherapy) higher level of carbonyl groups than in control healthy group. The level of 3-nitrotyrosine in platelet proteins from patients with invasive breast cancer was also significantly higher than in healthy subject group. We observed an increase of other biomarkers of oxidative stress such as O(2)(-·) and a decrease of GSH in platelets from patients with breast cancer (after the surgery and after various phases of the chemotherapy) compared to the healthy group. In model system in vitro our results showed that the commercial extract from berries of A. melanocarpa due to antioxidant action, significantly reduced the oxidative/nitrative stress in platelets from patients with invasive breast cancer caused by the surgery and various phases of the chemotherapy.     

Phagocytosis of neutrophils in rabbits infected with antigenic variants of RHD (rabbit haemorrhagic disease) virus

The present study was aimed at determining changes in chosen elements of phagocytosis in rabbits infected with 3 antigenic variants of RHD - Hartmannsdorf, Pv97 and 9905, which differed in haemagglutination ability. The animals were tested for phagocytosis parameters, and the results revealed that the examined strains showed the differences. These variations regarded mainly Pv97 strain, as the intensity of the changes were 5 times stronger in comparison to strain Hartmannsdorf and 9905. As all of the strains examined are signified as antigenic variants, we have stated that this feature does not determine their immunological picture. The results suggest the existence of immunological dissimilarities among strains of the RHD virus, which was revealed for the first time in antigenic variants.     

Assessment of treatment of Osteochondrosis dissecans (OCD) of shoulder joint in dogs--the results of two years of experience

Osteochondrosis is a common and clinically important joint disorder that occurs in human and many species of animals such as pigs, horses and dogs. The aim of this article is to present the results of conservative and surgical treatment of Osteochondrosis dissecans (OCD) and to compare the recovery time in relation to race, age and extent of cartilage damage. The study was performed on 36 dogs of both sexes, aged from 4 to 24 months, weighing 12-70 kg, of different breeds (predisposed and non-specific for this disease). All patients underwent clinical, orthopedic and radiological examination through which the location of OCD changes in the shoulder joint and the degree of this problem were established. Based on the examinations the patients were qualified for particular treatment procedures. Control tests were performed after 14, 30 and 60 days, which allowed for a precise evaluation of the progress of therapy. Achieved results indicate that dogs age and the lesions extent in the shoulder joint have an impact on the outcome of treatment. Studies showed that the best therapeutic effects were obtained in dogs aged 6-10 months, which underwent surgery. The study proved that the early diagnosis of OCD strongly influence the recovery rate and may inhibit the development of degenerative changes in the joint (DJD).