Extracardiac adult-type rhabdomyoma-Report of two cases in dogs.

This report describes the cytologic, histopathologic, and immunohistochemical features of adult-type rhabdomyoma located within the subcutaneous tissue in a 14-year-old female Border Collie (thigh) and a 13-year-old male Mongrel (flank). In both cases, fine-needle aspiration biopsy revealed cluster-forming, epithelial-like polygonal cells with abundant foamy cytoplasm, and moderate to marked anisocytosis and anisokaryosis; therefore, an epithelial tumor was suspected. After surgical excision, tumors underwent histopathologic examination with additional immunohistochemistry. Both tumors were well-demarcated and located within the subcutaneous tissue in the vicinity of the cutaneous muscle. The tumor mass consisted of densely packed round or polygonal cells with distinct vacuolation of the cytoplasm. Tumor cells expressed vimentin, desmin, and NSE and were cytokeratin and α-SMA negative. Based on histologic features and immunophenotyping, adult-type rhabdomyoma was diagnosed in both cases. This study highlights that the cytologic features of rhabdomyoma can be misleading and may suggest an epithelial tumor.

Influence of deoxynivalenol and zearalenone on the immunohistochemical expression of oestrogen receptors and liver enzyme genes in vivo in prepubertal gilts.

Deoxynivalenol (DON) and zearalenone (ZEN) are often detected in plant materials used to produce feed for pre-pubertal gilts. Daily exposure to small amounts of these mycotoxins causes subclinical conditions in pigs and affects various biological processes (e.g. mycotoxin biotransformation). The aim of this preclinical study was to evaluate the effect of low monotonic doses of DON and ZEN (12 µg/kg body weight-BW-and 40 µg/kg BW, respectively), administered alone or in combination to 36 prepubertal gilts for 42 days, on the degree of immunohistochemical expression of oestrogen receptors (ERs) in the liver and the mRNA expression of genes encoding selected liver enzymes during biotransformation processes. The level of expression of the analysed genes proves that the tested mycotoxins exhibit variable biological activity at different stages of biotransformation. The biological activity of low doses of mycotoxins determines their metabolic activity. Therefore, taking into account the impact of low doses of mycotoxins on energy-intensive processes and their endogenous metabolism, it seems that the observed situation may lead to the activation of adaptation mechanisms.

Tamoxifen decreases ovarian toxicity without compromising cancer treatment in a rat model of mammary cancer.

BACKGROUND: Premenopausal women diagnosed with breast cancer often face aggressive chemotherapy resulting in infertility. Tamoxifen (TAM) is a selective estrogen receptor modulator that was previously suggested as a protective agent against chemotherapy-induced ovarian failure. In the current study, we examined mechanisms of the protective action of TAM in the ovaries of tumor-bearing rats treated with the chemotherapy drug cyclophosphamide (CPA). RESULTS: TAM prevented CPA-induced loss of ovarian follicular reserves. The protective TAM effect in the rat ovary partially resulted from decreased apoptosis. In addition, transcriptomic and proteomic screening also implicated the importance of DNA repair pathways as well as cell adhesion and extracellular matrix remodeling in the protective ovarian actions of TAM. CONCLUSIONS: Tamoxifen shielded the ovary from the side effects of chemotherapy without lessening the tumoricidal actions of mammary cancer treatment.

Myofibroblastic tumour in the liver of a cat - A case report.

This study presents a case of a primary hepatic myofibroblastic tumour in a 15-year-old European Shorthair female cat. The cat showed a gradual increase in liver enzymes (alanine aminotransferase and aspartate aminotransferase), and an abdominal ultrasound revealed a tumour located within the left lateral lobe of the liver. The tumour was surgically excised and sent for histopathology. Histopathological examination showed that the tumour was composed of homogeneous fusiform cells with low mitotic count, crowded within the perisinusoidal, portal and interlobular spaces, and entrapment of hepatocytes and bile ducts. Immunohistochemistry revealed that the tumour cells expressed vimentin and α-SMA, and were negative to desmin and cytokeratins. Based on the histological and immunohistochemical features, as well as some similarities with analogous entities in humans and animals, the tumour was classified as a myofibroblastic neoplasm originating from the liver.

Metallothionein expression in feline injection site fibrosarcomas.

BACKGROUND: Feline injection site fibrosarcoma is an aggressive and infiltrative tumour arising in the background of chronic inflammation. The aim of this study was to evaluate the expression of metallothionein (I-II) in feline injection site fibrosarcomas and to assess its possible relationships with Ki67 index, inflammation score and tumour grade. The study included 40 feline fibrosarcomas, located in the common injection sites (i.e., interscapular area, thigh, flank), constituting archival diagnostic specimens collected between 2019-2020. Tumours were graded histologically according to the newly proposed soft-tissue sarcoma grading system in cats. Immunohistochemistry was performed to evaluate the expression of Ki67 and metallothionein in tumour cells. RESULTS: The cytoplasmic and sometimes nuclear expression of metallothionein was observed in all tumours grade I, 66.67% of tumours grade II and 55% of tumours grade III. The expression of metallothionein was negatively correlated with tumour grade and inflammation score, while the Ki67 index was positively correlated with tumour grade, inflammation score and necrosis score. CONCLUSION: The downregulation of MT expression in feline injection site fibrosarcomas seems to be connected with an increase in the inflammatory infiltration, hence tumour progression. This is the first study describing metallothionein expression in feline injection site fibrosarcomas.

Immunohistochemical Expression (IE) of Oestrogen Receptors in the Intestines of Prepubertal Gilts Exposed to Zearalenone.

This study was conducted to determine if a low monotonic dose of zearalenone (ZEN) affects the immunohistochemical expression (IE) of oestrogen receptor alpha (ERα) and oestrogen receptor beta (ERß) in the intestines of sexually immature gilts. Group C (control group; n = 18) gilts were given a placebo. Group E (experimental group; n = 18) gilts were dosed orally with 40 µg ZEN /kg body weight (BW), each day before morning feeding. Samples of intestinal tissue were collected post-mortem six times. The samples were stained to analyse the IE of ERα and Erß in the scanned slides. The strongest response was observed in ERα in the duodenum (90.387-average % of cells with ERα expression) and in ERß in the descending colon (84.329-average % of cells with ERß expression); the opposite response was recorded in the caecum (2.484-average % of cells with ERα expression) and the ascending colon (2.448-average % of cells with ERα expression); on the first two dates of exposure, the digestive tract had to adapt to ZEN in feed. The results of this study, supported by a mechanistic interpretation of previous research findings, suggest that ZEN performs numerous functions in the digestive tract.

Cutaneous and Subcutaneous Tumours of Small Pet Mammals-Retrospective Study of 256 Cases (2014-2021).

Since small mammals are gaining popularity as pets in Poland, the number of tumour samples submitted for histopathological examination is quite high. This study was a retrospective analysis of cutaneous and subcutaneous tumours in small pet mammals submitted for histopathology in 2014-2021. The analysis included 256 tumours sampled from 103 guinea pigs, 53 rats, 43 pet rabbits, 21 ferrets, 17 hamsters, 8 degus, 5 African pygmy hedgehogs, 3 Mongolian gerbils and 3 chinchillas. Tumours were diagnosed based on routine histopathology, with additional immunohistochemistry when necessary. The results of this study revealed that the vast majority of cutaneous tumours in guinea pigs were benign, with a predominance of lipoma. Adnexal tumours constituted a significant percentage of cutaneous tumours in guinea pigs (24.3%, with the most common being trichofolliculoma), pet rabbits (46.5%, with the most common being trichoblastoma), ferrets (33.3%, mostly derived from sebaceous glands), hamsters (52.9%, with the most common being trichoepithelioma) and gerbils (66.7%, scent gland epithelioma). Soft tissue sarcomas were a predominant group of tumours in rats (52.8%, with the most common being fibrosarcoma), African pygmy hedgehogs (100%), degus (87.5%) and chinchillas (66.7%). Melanocytic tumours were only sporadically seen in small mammal pets. Mast cell tumours were diagnosed only in ferrets, while epitheliotropic T-cell lymphoma was diagnosed only in a hamster and a degu. In summary, malignant tumours constitute a significant percentage of cutaneous tumours in many species of small mammal pets. Therefore, each cutaneous tumour should be sampled for further cytologic or histopathologic diagnosis.

The Effect of 42-Day Exposure to a Low Deoxynivalenol Dose on the Immunohistochemical Expression of Intestinal ERs and the Activation of CYP1A1 and GSTP1 Genes in the Large Intestine of Pre-pubertal Gilts.

Deoxynivalenol (DON) is a mycotoxin that contaminates various plant materials. Exposure to DON can disrupt hormonal homeostasis, decrease body weight gains and modulate the immune system in pigs. It can also cause diarrhea, vomiting, leukocytosis, hemorrhaging or even death. Prolonged exposure to low doses of DON can have serious health implications in mammals. This is the first in vivo study to show that per os administration of low DON doses probably contributes to specific dysfunctions in steroidogenesis processes by inducing the immunohistochemical expression of estrogen receptors alpha (ERα) in the entire gastrointestinal tract in strongly stained cells (3 points) and estrogen receptors beta (ERß), but only in both investigated segments of the duodenum in pre-pubertal gilts. Therefore, the aim of this study was to determine whether a NOAEL dose of DON (12 µg DON/kg BW) administered per os over a period of 42 days induces changes in the immunohistochemical expression of ER in different intestinal segments and the transcriptional activation of CYP1A1 and GSTP1 genes in the large intestine of pre-pubertal gilts. This is the first report to demonstrate the expression of ER, in particular ERß, with the associated consequences. The expression of ER was accompanied by considerable variations in the activation of CYP1A1 and GSTP1 genes, but it supported the maintenance of a stable consensus between the degree of mycotoxin exposure and the detoxifying effect in pre-pubertal gilts.

Equine Sarcoids-Causes, Molecular Changes, and Clinicopathologic Features: A Review.

Equine sarcoid is the most common skin tumor of horses. Clinically, it occurs as a locally invasive, fibroblastic, wart-like lesion of equine skin, which has 6 clinical classes: occult, verrucose, nodular, fibroblastic, mixed, and malignant. Sarcoids may be single but multiple lesions are more frequent. The typical histological feature is increased density of dermal fibroblasts which form interlacing bundles and whorls within the dermis. Lesions are mostly persistent, resist therapy, and tend to recur following treatment. In general, sarcoids are not fatal but their location, size, and progression to the more aggressive form may lead to the withdrawal of a horse from use and serious infringement of their welfare leading to the loss of valuable animals. Bovine papillomavirus (BPV) type 1 and less commonly type 2 contribute to the development of equine sarcoid. The viral genome and proteins are detected in a high percentage of cases. Furthermore, viral oncoprotein activity leads to changes in the fibroblastic tissue similar to changes seen in other types of tumors. Equine sarcoids are characterized by a loss of tumor suppressor activity and changes allowing abnormal formation of the affected tissue, as well as y immune defense abnormalities that weaken the host's immune response. This impaired immune response to BPV infection appears to be crucial for the development of lesions that do not spontaneously regress, as occurs in BPV-infected cows.

Mutations of p53 Gene in Canine Sweat Gland Carcinomas Probably Associated with UV Radiation.

INTRODUCTION: Apocrine sweat gland carcinomas (ASGCs) are rare malignant skin tumours in dogs and humans. The literature published so far focuses mostly on the clinico-epidemiological aspect of these tumours, but little is known about their pathogenesis. In this study we aimed to determine whether the p53 gene is involved in the carcinogenesis of the apocrine sweat gland in dogs and whether ultraviolet radiation (UV) is related to it. MATERIAL AND METHODS: Forty canine ASGCs were submitted to laser capture microdissection to isolate neoplastic cells, from which DNA was subsequently extracted. PCR amplification and sequencing of p53 exons 2-8 was then performed, followed by computer analysis of the obtained sequences. RESULTS: Sixteen mutations within the p53 gene were found in 13 tumours. The mutations involved C → T, T → C, G → A, and CC → TT transitions, C → G transversion and adenine deletion, which are gene alteration types known to be related to UV radiation in the process of skin carcinogenesis in humans. Six of the thirteen tumour cases displayed the C → T transitions in the same location in exon 4 and three of the thirteen cases displayed T → C in the same location in exon 5. CONCLUSION: The results of the present study indicate both the participation of the p53 gene and the influence of UV radiation in the formation of ASGCs in dogs.

Two different clinical forms of squamous cell carcinoma (SCC) in the perineum and vulva of two mares.

BACKGROUND: Genital malignant neoplasms in mares are relatively rare. The treatment involve surgical removal of the tumour masses, chemotherapy or both. CASE PRESENTATION: Two elderly warmblood mares, aged 16 and 20 were presented in University Clinic with the lumpy lesions at the region of perineum and left labia. Surgical removals of tumour masses were performed on standing animals. Removed tissues were subjected to histopathological examination which confirmed SCC. CONCLUSIONS: Clinical and ultrasound examination of reproductive organs in both mares showed no inflammatory or neoplastic changes. Both mares healed within 2 weeks after surgery and showed no signs of tumour recurrence for the following year despite no chemotherapy treatment.

Correlations between exposure to deoxynivalenol and zearalenone and the immunohistochemical expression of estrogen receptors in the intestinal epithelium and the mRNA expression of selected colonic enzymes in pre-pubertal gilts.

Plant-based materials used in the production of pig feed are very often contaminated with deoxynivalenol and zearalenone. Daily intake of small amounts of these mycotoxins with feed induces various subclinical states in gilts and influences different biological processes. The aim of this preclinical study was to determine the correlation between monotonic doses of zearalenone and deoxynivalenol (40 µg/kg body weight and 12 µg/kg body weight, respectively, administered over a period of 42 days) and the immunohistochemical expression of estrogen receptors in the intestinal tract and the mRNA expression of selected colonic enzymes. The immunohistochemical expression of estrogen receptor alpha was observed in the colon, but its intensity varied in different weeks of exposure. A minor increase in estrogen receptor beta expression was noted only in the colon, whereas the expression of cytochrome P450 1A1 enzyme mRNA and mRNA isoform of the glutathione S-transferase π gene decreased. The observed correlations suggest that the risk of loss of control over the biotransformation and biological activity of the parent compounds in distal intestinal mucosa is delayed.

Diagnostic immunohistochemistry for canine cutaneous round cell tumours - retrospective analysis of 60 cases.

INTRODUCTION: Canine cutaneous round cell tumours (CCRCTs) include various benign and malignant neoplastic processes. Due to their similar morphology, the diagnosis of CCRCTs based on histopathological examination alone can be challenging, often necessitating ancillary immunohistochemical (IHC) analysis. This study presents a retrospective analysis of CCRCTs. MATERIALS AND METHODS: This study includes 60 cases of CCRCTs, including 55 solitary and 5 multiple tumours, evaluated immunohistochemically using a basic antibody panel (MHCII, CD18, Iba1, CD3, CD79a, CD20 and mast cell tryptase) and, when appropriate, extended antibody panel (vimentin, desmin, a-SMA, S-100, melan-A and pan-keratin). Additionally, histochemical stainings (May-Grünwald-Giemsa and methyl green pyronine) were performed. RESULTS: IHC analysis using a basic antibody panel revealed 27 cases of histiocytoma, one case of histiocytic sarcoma, 18 cases of cutaneous lymphoma of either T-cell (CD3+) or B-cell (CD79a+) origin, 5 cases of plas-macytoma, and 4 cases of mast cell tumours. The extended antibody panel revealed 2 cases of alveolar rhabdo-myosarcoma, 2 cases of amelanotic melanoma, and one case of glomus tumour. CONCLUSIONS: Both canine cutaneous histiocytoma and cutaneous lymphoma should be considered at the beginning of differential diagnosis for CCRCTs. While most poorly differentiated CCRCTs can be diagnosed immunohis-tochemically using 1-4 basic antibodies, some require a broad antibody panel, including mesenchymal, epithelial, myogenic, and melanocytic markers. The expression of Iba1 is specific for canine cutaneous histiocytic tumours, and more sensitive than CD18. The utility of CD20 in the diagnosis of CCRCTs is limited.

Dirofilaria repens-An etiological factor or an incidental finding in cytologic and histopathologic biopsies from dogs.

Dirofilaria repens is an endemic, zoonotic parasite of carnivores, causing subcutaneous dirofilariasis, which is mostly asymptomatic. The aim of this study was to describe 22 cases of canine subcutaneous dirofilariasis. The cytologic and histopathologic samples were collected from dogs, which presented with various clinical signs such as cutaneous/subcutaneous nodules, hydropericardium, ascites, and lymphadenomegaly. The cytologic or histopathologic examination revealed purulent, pyogranulomatous, granulomatous or eosinophilic dermatitis/panniculitis, and the presence of D repens microfilariae or adults. The microfilariae or adults were also found incidentally in neoplastic cutaneous or subcutaneous tumors and in a sialocele. For the first time, microfilariae were also detected and described in pericardial and abdominal effusions and in enlarged reactive lymph nodes. Although it is well known that D repens can cause dermatitis and panniculitis in dogs, no previous reports of D repens microfilariae in body cavity fluids were found. The role of this parasite in the accumulation of body cavity fluid or in reactive lymphadenomegaly requires further investigation. Due to its zoonotic potential, and apparently underestimated pathogenicity, each case of canine subcutaneous dirofilariasis should be treated.

Cutaneous Leiomyosarcoma with Multiple Visceral Metastases in a Domestic Pigeon.

A clinical examination of a 4-yr-old fantail pigeon revealed a large skin tumor in the area of the left wing. The tumor had solid consistency and a single cavity filled with fluid. During necropsy, multiple tumors of varied size and diameter were found in the liver, spleen, pancreas, and kidneys. A histopathological examination of the skin tumor revealed extensive multifocal neoplasia with a tendency to infiltrate, composed of polygonal, round, oval, and elongated cells forming a solid system with cellular bands and obliteration of intercellular boundaries. Multifocal necrosis was detected in the foci of neoplastic infiltration. Similar foci were present in the liver, pancreas, and lungs. In an immunohistochemical examination, tumor cells tested positive for smooth-muscle actin and vimentin. The diagnosis was cutaneous leiomyosarcoma with an uncommon feature of visceral metastases.

Epidemiological and pathological features of primary cardiac tumours in dogs from Poland in 1970-2014.

Primary heart tumours affect less than 1% of dogs. Due to their rare incidence, every research showing the frequency of cardiac tumours is valuable. Routine diagnostics is often complemented with immunohistochemical analysis. This study was conducted on 110 patient records from all veterinary faculties in Poland from dogs diagnosed with heart tumours between 1970 and 2014. The dogs' age, breed and sex with tumour localisation and histopathological diagnosis were analysed. Because of its most common incidence, samples of haemangiosarcoma underwent further examination with assessment of the expression of cell markers that have not been evaluated earlier (i.e. minichromosome maintenance proteins and beta-catenin). We noted 111 tumours including 88.3% malignant and 10.8% benign ones. Haemangiosarcoma and aortic body tumour were the most frequent cardiac neoplasms in the dogs examined (45.9% and 27.9% of all tumours, respectively). Immunohistochemical analysis of haemangiosarcoma showed a positive expression of all markers examined. CD31, vimentin, and beta-catenin showed a positive reaction in all 11 samples examined. At least one proliferative marker (Ki-67, MCM-3 or MCM-7) showed a positive reaction in each sample. MCM-3 showed a higher expression than the two other proliferative markers (P = 0.006), but only Ki-67 showed a positive correlation with the mitotic index (P > 0.05, r = 0.89). Although beta-catenin, MCM-3 and MCM-7 showed a positive reaction in the haemangiosarcomas examined, their usefulness as diagnostic and prognostic factors should be a topic of further research.

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.

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.