Functional and morphological comparison of extracellular matrices derived from porcine small intestinal submucosa and porcine dermis to support rabbit uterus regeneration.

he aim of this study was to compare the ability of extracellular matrices (ECMs) of various structural origins to support uterus regeneration and maintain contractile function. The experiment was carried out in 18 New Zealand white female rabbits divided into a control group and two experimental groups. In the control group, bipolar electrodes were attached to the uterine horns to collect electromyography data. In the experimental groups, uterine horns were partially excised with immediate implantation of the extracellular matrix (ECM) derived from porcine small intestinal submucosa (SIS), DynaMatrix®, or from porcine dermis, Medeor® Matrix. Twelve weeks later, bipolar electrodes were attached to the regeneration areas in both experimental groups. The electromyography data in all groups were continuously recorded after administration of a single dose of estradiol valerate. Fourteen weeks after EMCs implantation, the rabbits were spayed, and the uterine wall was sampled for histological and immunohistochemical evaluation. The regenerated uterus retained the myoelectric activity pattern, and the mean amplitudes of maximum deflections did not differ between the experimental and control groups. The regeneration supported by the SIS-derived ECM was more advanced than that supported by the dermal-derived ECM. However, some pathological changes were noted in both experimental groups within the regeneration areas.

Intraocular Chondrosarcoma in a Rabbit.

Post-traumatic intraocular sarcomas are rarely reported in domestic animals and are most common in cats. An 8-year-old rabbit (Oryctolagus cuniculus) was referred to a veterinary clinic due to ocular discharge, uveitis and protein precipitate in the anterior chamber of the right eye. The eye was enucleated and histopathological examination revealed a poorly demarcated tumour within the ciliary body with invasion to adjacent eye structures. Neoplastic cells formed chaotic cartilage lacunae and were immunopositive for vimentin but immunonegative for pancytokeratin. On this basis, the neoplasm was diagnosed as a chondrosarcoma. To the best of our knowledge, this is the first report of intraocular chondrosarcoma in a rabbit. There was no history of previous ocular trauma but as there was serological evidence of Encephalitozoon cuniculi infection, inflammation could have been a predisposing factor to development of the neoplasm.

Canine and Feline Oral Cavity Tumours and Tumour-like Lesions: a Retrospective Study of 486 Cases (2015-2017).

Oral cavity tumours and tumour-like lesions are common in dogs and cats, and their diagnosis and classification requires histopathological examination. The aim of this study was to analyse retrospectively oral cavity lesions in dogs and cats in order to evaluate the distribution of inflammatory, hyperplastic and neoplastic lesions manifested as tumours. A total of 486 oral cavity tumours and tumour-like lesions (340 canine; 146 feline), diagnosed routinely from 2015 to 2017, were included. The lesions were classified as inflammatory, hyperplastic or neoplastic (benign and malignant). Histopathological diagnosis was based on haematoxylin and eosin staining and, when necessary, May-Grünwald-Giemsa (for mast cell tumours) or Masson's Fontana (for melanomas) stains or immunohistochemistry (for CD3, CD79α and S100 markers). For dogs, 29.11% (99/340) of the lesions were benign tumours, 24.12% (82/340) were hyperplastic lesions and 14.7% (50/340) were inflammatory lesions. For cats, 4.79% (7/146) were benign tumours, 15.07% (22/146) were hyperplastic lesions and 57.53% (84/146) were inflammatory lesions. Furthermore, 23.24% (79/340) of canine cases were diagnosed with gingival hyperplasia and 19.12% (65/340) were diagnosed with peripheral odontogenic fibroma, while 43.84% (64/146) of feline cases were diagnosed with chronic lymphoplasmacytic stomatitis. Malignant tumours in dogs and cats constituted 32.06% (109/340) and 21.91% (32/146) of the lesions, respectively, with high-grade melanoma in dogs and squamous cell carcinoma in cats being the most common.

Influence of simvastatin on hepatocytes - histopathological and immunohistochemical study.

The present study was undertaken to highlight the influence of simvastatin administration on hepatocyte morphology, proliferation, and apoptosis. The study included 48 gilts aged 3 months (weighing ca. 30 kg) divided into groups I (control; n=24) and II, receiving 40 mg/animal simvastatin orally (simavastatin; n=24) for 29 days. The animals were euthanized on days subsequent to the experiment. The livers were sampled, fixed, and processed routinely for histopathology, histochemistry, and immunohistochemistry (for proliferating cell nuclear antigen, Bcl-2, and caspase-3). Apoptosis was visualized by terminal deoxynucleotidyl transferase dUTP nick-end labelling (TUNEL). Simvastatin administration caused acute hepatocyte swelling, glycogen depletion, hyperaemia, multifocal hepatocyte proliferation with occasional pseudoacinar formation, connective tissue hyperplasia, eosinophil infiltration, and interface hepatitis. The proliferating cell nuclear antigen index, mean diameter of argyrophilic nucleolar organizer regions, and Bcl-2 immunoexpression were lower compared to control, and mean caspase-3 immunoexpression was higher in group II compared to control. On day 25 and 29 single hepatocytes in the simvastatin- treated group were TUNEL-positive. Simvastatin caused morphological alteration which became more intense over time. The results from the present study suggest that simvastatin treatment may cause glycogen, lipid metabolism and cell membrane permeability distortion, fibrosis, interface hepatitis, reduction in hepatocyte proliferation and transcriptional activity, and enhanced vulnerability to apoptosis. Summing up the results, it can be concluded that simvastatin caused liver damage with similar morphological changes seen in autoimmune-like liver injury, which may indicate that simvastatin may induce autoimmune-like drug induced liver injury.

Effects of caponisation and age on the histology of the internal organs of Leghorn cockerels.

1. Poultry researchers seek the most efficient bird line to produce capons (castrated cockerels). The previous studies did not include results from histopathological analyses of internal organs after caponisation. No data have been published on whether adipose tissue accumulates in internal organs or if caponisation changes the structure of the lymphoid organs and the accumulation of lymphoid cells. 2. The aim of this study was to analyse the occurrence of histopathological lesions in internal organs from Leghorn (layer-type) capons and cockerels at different times of fattening. 3. Two hundred, one-day-old Leghorn cockerels were used in this experiment. At 8 weeks of age, the birds were randomly divided into a control group (100 uncastrated cockerels) and a second group consisting of 100 castrated birds. At 12, 16, 20, 24 and 28 weeks of age, six cockerels and six capons were slaughtered. Samples of all internal organs from six cockerels and six capons at each slaughter age were evaluated. 4. The main histopathological differences between capons and cockerels were related to the accumulation of adipose tissue, with higher values noted in capons, especially in the gizzard and the caecum submucosa and fatty degeneration in liver hepatocytes. 5. Additionally, differences were observed more often in cockerels than in capons with regard to depletion of lymphoid cells in lymphoid organs, including the thymus and bursa of Fabricius in 28-week-old birds and the spleen in 24-week-old birds. 6. These studies prove that caponisation causes the accumulation of fat in internal organs and changes the structure of lymphoid organs. 7. Age influences the occurrence of desirable lesions, such as the accumulation of adipose tissue within the examined organs, and older capons (24 and 28 weeks old) are better sources of high quality, potentially edible tissues.

Major histocompatibility complex class I in the horse (Equus caballus) placenta during pregnancy and parturition.

BACKGROUND: Major histocompatibility protein class I (MHC-I) is believed to be expressed in the horse allantochorion only in limited areas at limited times. However, its expression has only been investigated in early pregnancy with non-quantitative techniques that cannot reliably detect small amounts of protein. OBJECTIVE: To quantify the relative expression of MHC-I in the allantochorion and endometrium during days 90-240 of pregnancy (PREG), parturition with physiological delivery of fetal membranes (PHYS), and parturition with retention of these membranes (FMR). Also, to visualize protein expression and determine whether classical or non-classical MHC-I mRNA is expressed. ANIMALS: Heavy draft horses. SETTING: PREG horses (n = 12) were sampled postmortem at a slaughterhouse. PHYS (n = 6) and FMR (n = 5) horses were sampled at farms in the vicinity of Olsztyn, Poland. METHODS: For relative quantification of MHC-I, western blotting with densitometry was used. To visualize MHC-I, immunohistochemistry was used. For mRNA identification, RT-PCR was performed. RESULTS: Although the quantity of MHC-I was lower during PREG than parturition, it was present in the allantochorion and endometrium during PREG. During parturition, MHC-I expression was upregulated in the allantochorion (PHYS vs. PREG: 2.7-times higher, 95% confidence interval, 1.3- to 5.7-times higher; FMR vs. PREG: 3.2-times higher, 95% confidence interval, 1.5- to 6.7-times higher). At parturition, staining for MHC-I was detected in the microcotyledons. Classical and non-classical MHC-I were expressed in both tissues during PREG, PHYS, and FMR. CONCLUSION: MHC-I protein is present in the horse allantochorion and endometrium for at least the first two-thirds of pregnancy and at parturition.

Morphology and immunoreactivity of canine and feline extramedullary plasmacytomas.

The aim of the study was the evaluation of morphology and immunophenotype of canine (19 cases) and feline (7 cases) extramedullary plasmacytomas. Tumours, located in skin, oral cavity and spleen were surgically excised, fixed and processed for histopathology and immunohistochemistry (CD79α, CD18, proliferating cell nuclear antigen, metallothionein). Histologically, tumours were classified into mature, cleaved, asynchronous, polymorphous blastic, hyalin, or monomorphous blastic type. All evaluated tumours showed cytoplasmic expression of CD79α antigen. The expression of CD18 was observed in canine cutaneous and splenic tumours. In canine tumours expression of metallothionein was low to moderate, while in feline plasmacytomas - absent or low. In canine tumours, the mitotic index and proliferating cell nuclear antigen index were positively correlated with the expression of metallothionein. In feline tumours no correlation between mitotic index, proliferating cell nuclear antigen and metallothionein was found. This is the first study describing expression of metallothionein in canine and feline extramedullary plasmacytoma.

Safety of the long-term application of QuikClot Combat Gauze, ChitoGauze PRO and Celox Gauze in a femoral artery injury model in swine - a preliminary study.

The purpose of this study was to examine the safety of the long-term application of QuikClot Combat Gauze, ChitoGauze PRO and Celox Gauze using a swine model. The study was conducted on nine pigs weighing approximately 30 kg, which were randomly divided into three groups. Under deep anesthesia, the pigs underwent complete transverse cutting of the femoral artery in the groin region. Hemostatic dressings were left in the wound for 24 hours. The animals were euthanized 24 hours after dressing application. In each group, macroscopic and microscopic severe changes and shock symptoms were observed in the lungs, liver, kidneys and heart. Fibrino-gaseous embolic material was found in the pulmonary artery of each group and in the lung vessels of the animals from the ChitoGauze PRO and Celox Gauze groups. In conclusion, the long-term application of the evaluated hemostatic dressings has the risk of coagulopathy and reaching the progressive stage of shock. The residues from the hemostatic dressings can ingress into the systemic circulation, thereby increasing the risk of embolus formation. Because of these harmful effects, the evaluated hemostatic dressings are not appropriate for long-term use. Future studies are needed on the consequences of the long-term application of these hemostatic agents.

Histopathological studies of the heart in three lines of broiler chickens.

The aim of this study was to analyse the morphological lesion pattern of the heart of broiler chickens (Cobb 500, Hubbard F15 and Ross 308) during fattening with no clinical signs of disease and to determine the most susceptible period for the occurrence of morphological lesions. The most frequently diagnosed lesions in each genetic line were degeneration of the fibres with vacuolation, congestion of cardiac muscle, oedema and vacuolisation of the Purkinje cells. The highest numbers of morphological lesions were observed on d 38, 31 and 10 of life. The lesions were most numerous in the septum, followed by the left and right ventricles. Ischaemic cardiomyocytes were also most numerous on d 38 of life and in the left ventricle. Overload of cardiac muscle, prolonged hypoxia and increasing body weight on d 38 are the likely reasons for the largest number of lesions and ischaemic fibres, which may lead to heart failure.

Effect of low-energy laser irradiation and antioxidant supplementation on cell apoptosis during skeletal muscle post-injury regeneration in pigs.

The aim of this study was to evaluate the effect of low-energy laser irradiation, coenzyme Q10 and vitamin E supplementation on the apoptosis of macrophages and muscle precursor cells during skeletal muscle regeneration after bupivacaine-induced injury. The experiment was conducted on 75 gilts, divided into 5 experimental groups: I--control, II--low-energy laser irradiation, III--coenzyme Q10, IV--coenzyme Q10 and vitamin E, V--vitamin E. Muscle necrosis was induced by injection of 0.5% bupivacaine hydrochloride. The animals were euthanized on subsequent days after injury. Samples were formalin fixed and processed routinely for histopathology. Apoptosis was detected using the TUNEL method. The obtained results indicate that low-energy laser irradiation has a beneficial effect on macrophages and muscle precursor cell activity during muscle post-injury regeneration and protects these cells against apoptosis. Vitamin E has a slightly lower protective effect, limited mainly to the macrophages. Coenzyme Q10 co-supplemented with vitamin E increases the activity of macrophages and muscle precursor cells, myotube and young muscle formation. Importantly, muscle precursor cells seem to be more sensitive to apoptosis than macrophages in the environment of regenerating damaged muscle.

Differences in extracellular matrix remodeling in the placenta of mares that retain fetal membranes and mares that deliver fetal membranes physiologically.

INTRODUCTION: In mammals, placenta separation at term may involve degradation of the extracellular matrix by matrix metalloproteinases (MMPs). The activity of MMPs is modulated by TIMPs. We hypothesized that the placentas of mares that deliver fetal membranes physiologically and those that retain fetal membranes (FMR) differ in terms of histology; mRNA expression of MMP-2 and MMP-9; protein expression of MMP-2, MMP-9, and TIMP-2; and the potential activity of both MMPs. METHODS: Placenta biopsies were taken from mares (n = 9; 4 FMR, 5 controls) immediately after foal expulsion. Retention was defined as failure to expel all fetal membranes within 3 h of expulsion. All mares were monitored for time of expulsion. The degree of allantochorial/endometrial adhesion was determined in FMR mares, and biopsies from all mares were histologically examined. mRNA expression, protein immunolocalization, protein amount and potential enzyme activity were determined with RT-PCR, immunohistochemistry, Western Blotting and zymography, respectively. RESULTS: FMR mares had strong to extremely strong allantochorial/endometrial adhesion, and significantly more connective tissue in the allantochorial villi than controls. The range of MMP-2 mRNA expression levels was more than 13 times greater in FMR mares than in controls. Protein content of both MMPs and TIMP-2 differed significantly between groups. The range of potential MMP-2 and MMP-9 activity was larger in FMR mares, and MMP-2 potential activity was 1.4 times higher in controls (P = 0.02). DISCUSSION: These results indicate differences in extracellular matrix remodeling in FMR mares and controls, and suggest dysregulation of MMP expression and activation in FMR mares.

Aggressive, solid variant of alveolar rhabdomyosarcoma with cutaneous involvement in a juvenile labrador retriever.

An 8.5-month-old male Labrador retriever presented with a cutaneous mass in the right maxillofacial region and swelling of the gingiva. The dog received antibiotic and anti-inflammatory treatment. After 3 weeks the dog returned, presenting with disseminated cutaneous tumours on the neck, trunk and groin. One of the nodules was resected and a cutaneous round cell tumour was diagnosed on microscopical examination. The dog was humanely destroyed. Necropsy examination revealed disseminated tumours in the skin, internal organs and skeletal muscles. Microscopically, all of the tumours were composed of small round cells, arranged in nests. Immunohistochemically, the neoplastic cells expressed vimentin, desmin, MyoD1, myogenin and smooth muscle actin, but were negative for CD3, CD18, CD79αcy, cytokeratin AE1/AE3, chromogranin A, class II molecules of the major histocompatibility complex, neuron-specific enolase and S100. The average Ki67 index was 89.5%. The final diagnosis was a solid variant of alveolar rhabdomyosarcoma (ARMS). This is the first report of the cutaneous multifocal form of ARMS in veterinary oncology.

Cytomorphometry of canine cutaneous histiocytoma.

A morphometric analysis of tumoral Langerhans cells and activated macrophages was conducted using canine cutaneous tumors (65 cases of canine cutaneous histiocytoma and 7 cases of pyogranuloma). The histiocytic origin of the tumor cells was confirmed using immunohistochemistry. The parameters of the morphometric analysis included cellular and nuclear size and shape and the nuclear:cytoplasmic ratio; the variability of these features was calculated separately for each tumor. The canine cutaneous histiocytoma group was divided into four stages of regression depending on the intensity of the lymphocytic infiltration. Statistical analysis revealed that the anisocytosis, anisokaryosis and cellular pleomorphism of tumoral Langerhans cells increased, while the cellular circularity and nuclear:cytoplasmic ratio decreased with tumor regression. Activated macrophages of the pyogranuloma were significantly larger, and had larger nuclei, than tumoral Langerhans cells. Furthermore, these activated macrophages showed greater anisocytosis and anisokaryosis and a lower nuclear:cytoplasmic ratio than tumoral Langerhans cells in the first stages of tumor regression. These results indicate that tumoral Langerhans cells undergo morphologic changes during the regression of canine cutaneous histiocytoma, reflecting their maturation and differentiation. Morphometry can be a useful method for distinguishing activated macrophages from tumoral Langerhans cells.

Immunohistochemical evaluation of superoxide dismutase (Cu/Zn SOD) concentrations in erythrocytes of dairy cattle and farm-raised deer by a computer-assisted analysis of microscopic images.

The effectiveness of the immunohistochemical method in determining Cu/Zn SOD concentrations in red blood cells of dairy cattle and farm-raised deer was evaluated by a computer-assisted analysis of microscopic images and scanning technique. Superoxide dismutase (Cu/Zn SOD) concentrations in erythrocytes were determined in smears of whole blood samples collected from 16 Polish Holstein-Friesian cows and 22 farm-raised deer in spring. Mouse anti-bovine SOD (Cu-Zn) monoclonal antibodies (2F5, Serotec) were used in 1:50 dilution. The degree of immunostaining for SOD in red blood cells was determined with the use of the MIDI 3DHistech Panoramic Scanner (Hungary) and 3DHistech Panoramic Viewer, NuclearQuant and MembraneQuant software. Our findings indicate that the immunohistochemical method is a useful technique for evaluating Cu/Zn SOD concentrations in red blood cells of cattle and deer.