Iwona Otrocka-Domagała

IκB kinase β inhibitor, IMD-0354, prevents allergic asthma in a mouse model through inhibition of CD4(+) effector T cell responses in the lung-draining mediastinal lymph nodes.

IκB kinase (IKK) is important for nuclear factor (NF)-κB activation under inflammatory conditions. It has been demonstrated that IMD-0354, i.e. a selective inhibitor of IKKβ, inhibited allergic inflammation in a mouse model of ovalbumin (OVA)-induced asthma. The present study attempts to shed light on the involvement of CD4(+) effector (Teff) and regulatory (Treg) T cells in the anti-asthmatic action of IMD-0354. The animals were divided into three groups: vehicle treated, PBS-sensitized/challenged mice (PBS group); vehicle treated, OVA-sensitized/challenged mice (OVA group); and IMD-0354-treated, OVA-sensitized/challenged mice. The analyzed parameters included the absolute counts of Treg cells (Foxp3(+)CD25(+)CD4(+)), activated Teff cells (Foxp3(-)CD25(+)CD4(+)) and resting T cells (CD25(-)CD4(+)) in the mediastinal lymph nodes (MLNs), lungs and peripheral blood. Moreover, lung histopathology was performed to evaluate lung inflammation. It was found that the absolute number of cells in all studied subsets was considerably increased in the MLNs and lungs of mice from OVA group as compared to PBS group. All of these effects were fully prevented by treatment with IMD-0354. Histopathological examination showed that treatment with IMD-0354 protected the lungs from OVA-induced allergic airway inflammation. Our results indicate that IMD-0354 exerts anti-asthmatic action, at least partially, by blocking the activation and clonal expansion of CD4(+) Teff cells in the MLNs, which, consequently, prevents infiltration of the lungs with activated CD4(+) Teff cells. The beneficial effects of IMD-0354 in a mouse model of asthma are not mediated through increased recruitment of Treg cells into the MLNs and lungs and/or local generation of inducible Treg cells.

Morphology and immunophenotype of canine cutaneous histiocytic tumours with particular emphasis on diagnostic application

This study evaluated the morphology and immunohistochemistry of 85 canine cutaneous histiocytic tumours. The tumours were classified morphologically as either canine cutaneous histiocytomas (71 tumours) or canine cutaneous histiocytic sarcomas (14 tumours). The immunohistochemical analysis was conducted on paraffin sections using an antibody panel (against MHCII, CD18, CD79αcy, CD3 and E-cadherin). Histochemical staining with toluidine blue and Gomori silver impregnation was also performed. A follow-up was conducted via surveys. The histiocytic origin of the tumour cells was confirmed in 65 of the canine cutaneous histiocytomas and in 4 of the canine cutaneous histiocytic sarcomas. The tumours that had been misdiagnosed as canine cutaneous histiocytomas included plasmacytomas, epitheliotropic T-cell lymphomas and undetermined entities. The tumours misdiagnosed as canine cutaneous histiocytic sarcomas included plasmacytomas and non-epitheliotropic T-cell lymphomas, but the majority of them remained undetermined. The canine cutaneous histiocytomas showed MHCII, CD18 and E-cadherin expression, but in several of the tumours, the expression of CD18 or E-cadherin was confirmed in only a small percentage of the tumour cells. The regressing canine cutaneous histiocytomas showed increased T- and B-lymphocyte infiltration, a decreased mitotic index, transport of the MHCII molecules from the cytoplasm to the cell membrane and loss of E-cadherin expression in the tumour cells. The canine cutaneous histiocytic sarcomas showed both high morphological diversity and expression of MHCII and CD18. Two of the evaluated histiocytic sarcomas also showed expression of E-cadherin. In conclusion, immunohistochemistry, including analysis of MHCII, CD18 and the lymphocytic markers CD3 and CD79, should be performed for the diagnosis of canine cutaneous histiocytic tumours. The expression of E-cadherin in canine cutaneous histiocytic sarcomas suggests an origin of the tumour cells among Langerhans cells.

Pathomorphological changes in the alimentary system of Japanese quails naturally infected with Eimeria tsunodai

Abstract The aim of this study was to investigate pathomorphological changes in the duodenum, jejunum, ileum, and caecum of Japanese quails naturally infected with Eimeria tsunodai. Samples of the intestines were collected from 6-week-old cockerels and hens before laying and from laying quails aged 12, 24, and 48 weeks. The tissue sections were stained with haematoxylin and eosin, PAS method according to McManus, and Ziehl-Neelsen method. Morphometric and morphological analyses of coccidia revealed that Eimeria tsunodai was responsible for the infection. Different developmental stages of the coccidia were observed almost exclusively in the caecum, and they led to the complete damage of caecal mucosa. The main changes observed in the caecum involved mucosal damage, atrophy of the folds and crypts, and mucosal desquamation. No significant inflammation changes were detected. The degree of damage to caecal mucosa increased with age and the greatest damage was noted at the age of 48 weeks. Eimeria tsunodai infections occur in Japanese quails during the entire laying period. Different developmental stages of coccidia are responsible for total damage to caecal mucosa.

Extragonadal teratoma in a domestic turkey (Meleagris gallopavo domestica)

This is the first report of a primary, spontaneous and, most probably, congenital teratoma in a domestic turkey, localized in front of the left eyeball. The unique localization allowed surgical excision of the tumour. The histopathological examination revealed that the tumour included structures derived from all three germ cell layers: ectoderm, mesoderm and endoderm (e.g. cartilaginous, osseous, haematopoietic, fibrous, nervous, glandular, squamous epithelial and smooth muscle tissues). The presence of epithelial cells as well as smooth muscle cells was confirmed using anti-cytokeratin and anti-desmin antibodies, respectively. The proliferative activity of the tumour cells was confirmed using proliferating cell nuclear antigen immunostaining. The other cases of teratoma in wild and domestic birds are reviewed briefly.

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.

Prostaglandin E2 exerts the proapoptotic and antiproliferative effects on bovine NK cells

The aim of this research was to determine whether prostaglandin E2 (PGE2) affects bovine NK cells in respect of their counts, apoptosis and proliferation, and if it does, then which of the PGE2 receptor (EP) subtype(s) mediate(s) these effects. We here report that long-term, but not short-term, exposure of bovine peripheral blood mononuclear cells to PGE2 at 10(-5)M, 10(-6)M and 10(-7)M, but not at 10(-8)M, caused a significant increase in the percentage of early apoptotic cells among NK cell subset. Moreover, PGE2 at 10(-5)M and 10(-6)M, but not at 10(-7)M and 10(-8)M, induced a considerable decrease in the absolute count of NK cells. The magnitude of these effects increased with an increasing concentration of PGE2. The blockade of EP1, EP2, EP3 and EP4 receptors did not prevent the PGE2-induced apoptosis and depletion of NK cells. The results suggest that the proapoptotic effect of PGE2 is secondary in character and the induction of this effect is not mediated through EP receptors. Furthermore, the studies demonstrated that PGE2 at 10(-5)M and 10(-6)M, but not at 10(-7)M and 10(-8)M, highly significantly reduced the percentage of proliferating NK cells. The EP1, EP1/2 and EP3 receptor antagonists were unable to block this effect significantly, whereas the selective blockade of EP4 receptors prevented the PGE2-induced inhibition of NK cells proliferation. These results indicate that PGE2 at certain concentrations may impair the proliferation of NK cells and this effect is mediated via the EP4 receptor.

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.

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.

Effects of caponization and age on the histology, lipid localization, and fiber diameter in muscles from Leghorn cockerels

ABSTRACT Poultry researchers seek general and native lines of birds for capon production. The primary aim is to find high‐quality meat with valuable sensory attributes. Castration decreases androgen levels and changes lipid metabolism, resulting in the accumulation of abdominal, subcutaneous, and intramuscular fat, which changes the flavor, texture, and tenderness of the meat. Histological analysis of selected skeletal muscles from Leghorn capons and cockerels was the main aim of the present study, along with examination of lipid localization in muscle tissue. We also assessed fiber type and diameter in the pectoral muscles (pectoralis major). The experiment was performed on 200 Leghorn cockerels; testes were removed at 8 wk of age. At 12, 16, 20, 24, and 28 wk of age, 6 cockerels and 6 capons were slaughtered, and samples from the pectoral and thigh muscles were evaluated. Differences in the focal necrosis of the fibers with infiltration of lymphoid cells (P < 0.05) were observed in thigh muscles, with higher numbers in cockerels than in capons. All examined locations in the pectoral and thigh muscles of capons (around the blood vessels, in the perimysium, in the endomysium, and in the sarcoplasm) showed higher concentrations of lipids compared to levels in cockerels. The analysis of fiber type in pectoralis major muscles revealed that all fibers were type IIB. The diameters of the fibers of the pectoral muscles were different (P < 0.05) at 28 wk of age, and the diameters of the giant fibers were different (P < 0.05) at 16 wk of age. The high concentration of lipids in the skeletal muscles of Leghorn capons is remarkable. Thus, ethical reasons as well as economic reasons should be considered when one‐day‐old cockerels are eliminated.

Influence of long-term, high-dose dexamethasone administration on proliferation and apoptosis in porcine hepatocytes

The aim of this study was to examine the influence of long-term, high-dose dexamethasone administration on the liver, with particular emphasis on hepatocyte proliferation and apoptosis, using a swine model. The study included 48 large, female Polish breed pigs aged 3months (weighing ca. 30kg) divided into groups I (control; n=24) and II (dexamethasone; n=24) that receiving intra-muscular injections of monosodium phosphate dexamethasone for 29days. The pigs were euthanized on days subsequent to the experiment. Immediately after the euthanasia, the pig 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). Dexamethasone administration gradually caused hepatocyte glycogen degeneration and finally lipid degeneration, accompanied by sinusoid and central vein dilatation and nuclear chromatin condensation. The proliferating cell nuclear antigen index, mean number of argyrophilic nucleolar organizer regions and proliferation index of argyrophilic nucleolar organizer regions were lower, while Bcl-2 expression was higher in group II compared with group I. The results from this study suggest that safe high-dose dexamethasone administration time is difficult to establish. Long-term, high-dose dexamethasone administration can cause pronounced morphological changes in hepatocytes by diminishing their transcriptional and proliferation activity but also protects them from apoptosis by potentially affecting Bcl-2 expression.