Canine and feline in situ mammary carcinoma: A comparative review.

Carcinoma in situ of the breast is a well-known entity in humans. In veterinary medicine, particularly in canine and feline mammary literature, there is no agreement whether the term in situ should be used to indicate a specific carcinoma histotype or the noninvasive status of a carcinoma of any histotype. Moreover, in the most recent histologic classification of mammary tumors published by the Davis-Thompson Foundation, it is suggested to abandon the term carcinoma in situ given the lack of standardized criteria defining this entity, replacing it with epitheliosis or ductal/lobular hyperplasia with severe atypia. This publication presents a critical review of the term in situ in human and veterinary medicine considering the evolution of the term over the years and its heterogeneous use by different authors, including variations in immunohistochemical markers for classification. This review aims to point out the lack of uniformity in the nomenclature and classification issues in veterinary medicine regarding the use of the term in situ, laying the ground for a process of standardization in future publications.

Bovine besnoitiosis in a cattle herd in Sicily: an isolated outbreak or the acknowledgment of an endemicity?

Bovine besnoitiosis is a debilitating infectious disease caused by Besnoitia besnoiti (Apicomplexa; Sarcocystidae). The disease is mainly characterized by cutaneous and systemic signs, infertility in bulls, and abortion in cows. The current study describes an autochthonous outbreak of bovine besnoitiosis in Sicily, Southern Italy, being the first report of B. besnoiti infection in the island so far. In a cattle farm located in Syracuse province, a 4-year-old Belgian blue bull born in Sicily displayed typical clinical signs of the sub-acute/chronic disease phase with thickening of the skin of the scrotum and testicular degeneration. Histological examination of scrotal biopsies revealed the presence of several tissue cysts of B. besnoiti. The serological analysis of the herd using a commercial ELISA revealed a high seroprevalence (45 out of 55; 82%) of antibodies against B. besnoiti. Few seropositive animals (5 out of 45; 11%) showed clinical signs, cysts in vestibulum vaginae (1 out of 31; 3.2%), and testicular degeneration (4 out of 14; 28.6%) assessed by ultrasonographic investigations. The paucity of clinical signs associated with the high seroprevalence in the farm led to hypothesize that bovine besnoitiosis is endemic in the area though further studies are needed. Local practitioners should be more aware of the disease to facilitate the early detection of cases, prevent the spread of infection, and avoid economic losses and animal health problem.

Reaction of Cornu aspersum Immune System against Different Aelurostrongylus abstrusus Developmental Stages.

Cornu aspersum, the land snail, is recognized as a suitable intermediate host of Aelurostrongylus abstrusus; however, there is little information both on larval development as well as on the intermediate host's immune system reaction to the parasite. The aim of the study was to assess the histological reaction of C. aspersum's immune system against A. abstrusus. Sixty-five snails were provided by a snail farm. Five of them were digested to assess the absence of natural parasitic infections. The remaining sixty were divided into five groups. Three groups of snails were infected with A. abstrusus using by-contact or injection methods; one group was injected only with saline solution and one group was left untreated as the control. The snails of group A were sacrificed and digested on study days 2, 10, and 18; snails of the other groups were collected and examined for histopathological analysis on study days 2, 10, and 18. On study day 2, in the infected snails, several free L1s were observed along with the absence of immune system reactions. On day 10, the L2s elicited an intense reaction in the internal layer of the muscular foot. On day 18, all L3s partially encapsulated by the snail's immune system were observed in the outermost part of the muscular foot, which is near and among the goblet cells. This last finding suggests that L3s could be shed with the snail's mucus and spread in the environment, representing an alternative route of transmission for this feline lungworm.

Hemosiderin-Laden Macrophages in Canine Mammary Carcinomas.

Macrophages are among the main actors in cancer immunoediting, with several functions, including recycling iron and packaging it in hemosiderin. Even though TAMs are widely studied in breast cancer and canine mammary tumors, hemosiderin-laden macrophages (HLMs) have not received as much attention. Considering the growing interest in iron metabolism in cancer, this study aims to evaluate the presence of HLMs in canine mammary tumors. Fifty cases of canine mammary carcinomas presenting aggregates of pigmented macrophages were chosen. Prussian blue and Meguro staining were performed to assess the presence of iron. Immunohistochemistry was carried out to try to identify macrophagic phenotypes and hypothesize their role. Evaluation of the H&E sections showed that pigmented macrophages were variously localized in peritumoral and stromal areas. These pigmented cells were variably stained with Prussian blue and reacted strongly with DAB in the Meguro staining method, thus confirming the presence of iron within them. In their immunohistochemistry, the HLMs were negative for the MAC387 but positive for CD 204 and VEGF. Considering their positivity for CD 204, HLMs could be M2 macrophages that supply iron to both the neoplastic cells and the tumor inflammatory microenvironment, promoting angiogenesis and protecting cancer cells from hypoxia.

Prenatal Diagnosis of Canine and Feline Twins Using Ultrasound: A Retrospective Study.

Prenatal diagnosis comprises a set of investigations, both instrumental and laboratory-based, which aim to monitor the health of the foetus during pregnancy, from the early stages of embryonic development to the moments preceding delivery. A growing interest is emerging for the preterm ultrasound morphological screening of embryos and foetuses, aimed at assessing the integrity and viability of the conceptus, as well as the early diagnosis of anomalies which can cause complications. This study is a retrospective study of the ultrasonographic findings of twins in the authors' clinical activity from 2016 to 2022. Only seven cases of monochorionic twins were recorded, out of the whole number of evaluations performed on 3120 foetuses, with a prevalence of 0.6% and 0.2% in feline and canine foetuses. All the twins had their own amniotic sac and umbilical cord but presented a single placenta and a single allantoic sac. Unfortunately, the three feline cases were not more recognizable at term. In the four canine cases, three were of opposite sex and then necessarily dizygotic. Twins may have an impact on the success of a pregnancy due to the risk of dystocia, as observed in some of the reported cases. Prenatal ultrasound allows early recognition of twins in dogs and cats.

Beta-catenin expression in pilomatrix carcinoma with multiple visceral metastases in a dog.

Beta-catenin is a protein initially identified as a submembrane component of the E-cadherin-mediated cell-to-cell adhesion system. It plays a role as a transcriptional factor in the wingless/Wnt signalling pathway. Beta-catenin has been associated with oncogenic activity in human benign and malignant pilomatrix neoplasms where the immunohistochemical profile of ß-catenin expression displayed both nuclear and cytoplasmic staining in basaloid cells. In this study, an 8-year-old female Irish setter dog was examined because of the presence of skin nodules. Tissue biopsies from different nodules were obtained and histological examination suggested a diagnosis of pilomatrix carcinoma. The dog spontaneously died after 2 months and necropsy showed multiple metastases in the nasal cavity, lungs, heart, kidney, liver and colon. Routine histopathology of metastatic sites showed features consistent with the pattern of primary neoplastic nodules. Immunohistochemical detection of ß-catenin was performed in both the primary tumour and the metastases. Beta-catenin expression was located in the nuclei, cytoplasm and membrane of squamoid cells and in the cytoplasm of basaloid cells, while shadow cells were completely negative. To the best of our knowledge, these data represent the first report on the immunohistochemical expression profile of ß-catenin in canine pilomatrix carcinoma.

Does TLS Exist in Canine Mammary Gland Tumours? Preliminary Results in Simple Carcinomas.

Neoplastic progression is influenced by the expression of tumour antigens that activate an anti-tumour immune response. Human medical studies show that this body defence is carried out in secondary lymphoid organs (SLOs) but also directly in the tumour through organized cellular aggregates that are called tertiary lymphoid structures (TLSs). However, their occurrence has different meanings in different tumour types. For example, the presence of TLSs in breast cancer is associated with the most aggressive subtypes. This paper aimed to study TLSs in canine mammary simple carcinomas. A morphological assessment of the inflammatory infiltrate was performed on H&E sections of fifty cases. Immunohistochemistry was then carried out to typify the inflammatory cells in the tumour microenvironment. Results showed that, sometimes, inflammatory infiltrates were organized in follicles close to high-grade carcinomas, simulating a lymphoid organization, as in breast cancer. Therefore, we can assume that even in canine mammary tumours, TLSs exist and they are entities to consider due to their presence in the most aggressive histotypes or tumours with a high degree of malignancy.

Nontraumatic Paraureteral Urinoma in a Cat with Urolithiasis.

Urinoma is an encapsulated collection of urine due to a disruption in the collection system of the urinary tract. This condition is rarely reported in veterinary medicine. The aim of this study is to describe the clinical and diagnostic findings of a well-encapsulated paraureteral urinoma associated with urinary tract stones in a critical feline patient. The ultrasound examination of the abdomen revealed a well-defined collection of anechoic fluid containing the left kidney in the retroperitoneal space, while the radiographic examination revealed a loss of soft tissue contrast in the lumbar region. Both techniques confirmed the presence of stones in the bladder. Following the ultrasound-guided drainage of the lesion, the nature of the fluid was also confirmed to be urine. Due to the worsening of the cat's health, to better define the extent and genesis of the lesion, a positive contrast radiographic study was performed by means of an ultrasound-guided percutaneous injection of a contrast medium into the lesion, which highlighted a wide and homogeneous radiopaque area in the left retroperitoneal space. These findings were confirmed with a postmortem examination, corroborating the diagnostic suspicion of urinoma. The percutaneous contrast inoculation performed in this critically ill patient plays a role in the diagnostic process to reach a final diagnosis in cases in which the patient's clinical condition does not allow for performing an excretory positive contrast study.

Reproducibility and Feasibility of Classification and National Guidelines for Histological Diagnosis of Canine Mammary Gland Tumours: A Multi-Institutional Ring Study.

Histological diagnosis of Canine Mammary Tumours (CMTs) provides the basis for proper treatment and follow-up. Nowadays, its accuracy is poorly understood and variable interpretation of histological criteria leads to a lack of standardisation and impossibility to compare studies. This study aimed to quantify the reproducibility of histological diagnosis and grading in CMTs. A blinded ring test on 36 CMTs was performed by 15 veterinary pathologists with different levels of education, after discussion of critical points on the Davis-Thompson Foundation Classification and providing consensus guidelines. Kappa statistics were used to compare the interobserver variability. The overall concordance rate of diagnostic interpretations of WP on identification of hyperplasia-dysplasia/benign/malignant lesions showed a substantial agreement (average k ranging from 0.66 to 0.82, with a k-combined of 0.76). Instead, outcomes on ICD-O-3.2 morphological code /diagnosis of histotype had only a moderate agreement (average k ranging from 0.44 and 0.64, with a k-combined of 0.54). The results demonstrated that standardised classification and consensus guidelines can produce moderate to substantial agreement; however, further efforts are needed to increase this agreement in distinguishing benign versus malignant lesions and in histological grading.

Penicilliosis in an African grey parrot (Psittacus erithacus).

Penicillium mycosis, or penicilliosis, is rare in animals. This report describes the clinical signs and pathological, immunohistochemical, and biomolecular findings in a case of avian penicilliosis in a Congo African grey parrot. On gross necropsy, pyogranulomatous changes were identified in the lung, liver, and kidney. Histopathology demonstrated periodic acid Schiff-(PAS) and Grocott-positive hyphae in these tissues. Immunohistochemistry confirmed the genus of the fungus and polymerase chain reaction (PCR) confirmed a high homology with Penicillium chrysogenum.

Clinical Significance of ROMs, OXY, SHp and HMGB-1 in Canine Leishmaniosis.

This study aimed to investigate the role of oxidative stress parameters (ROMs, OXY, SHp), the Oxidative Stress index (OSi), and High Mobility Group Box-1 protein (HMGB-1) in canine leishmaniosis (CanL). For this study, thirty dogs, naturally infected with Leishmania spp. (Leishmania Group, LEISH) and ten healthy adult dogs (control group, CTR) were included. The diagnosis of CanL was performed by a cytological examination of lymph nodes, real time polymerase chain reaction on biological tissues (lymph nodes and whole blood), and an immunofluorescence antibody test (IFAT) for the detection of anti-Leishmania antibodies associated with clinical signs such as dermatitis, lymphadenopathy, onychogryphosis, weight loss, cachexia, lameness, conjunctivitis, epistaxis, and hepatosplenomegaly. The HMGB-1 and oxidative stress parameters of the LEISH Group were compared with the values recorded in the CTR group (Mann Whitney Test, p < 0.05). Spearman rank correlation was applied to evaluate the correlation between the HMGB-1, oxidative stress biomarkers, hematological and biochemical parameters in the LEISH Group. Results showed statistically significant higher values of SHp in the LEISH Group. Specific correlation between the ROMs and the number of red blood cells, and between HGMB-1 and SHp were recorded. These preliminary data may suggest the potential role of oxidative stress in the pathogenesis of CanL. Further studies are undoubtedly required to evaluate the direct correlation between inflammation parameters with the different stages of CanL. Similarly, further research should investigate the role of ROMs in the onset of anemia.

Pseudo-Placentational Endometrial Hyperplasia in the Bitch: Case Series.

Canine pseudo-placentational endometrial hyperplasia differs from the classical form of cystic endometrial hyperplasia for the well-organized tissue architecture resembling the canine placenta. After the discovery, it has been inconstantly reported. The present work reports the clinicopathological details of six spontaneous cases retrieved retrospectively from a large database. The lesion was found in young non-pregnant female dogs (median 2.0 years) at the end of dioestrus. It could be imaged by ultrasound and was always grossly detectable as single or multiple uterine enlargements of 2-3 cm in diameter with a villous whitish tissue growing on the mucosa and occluding the lumen. Histology confirmed the tissue architecture of the canine placenta with a basal glandular layer, a connective band, a spongy layer and a tortuous and compact labyrinth, often poorly recognizable. The pseudo-placentational hyperplasia is a non-inflammatory proliferative lesion although numerous mast cells inhabit the connective band, and a superimposed inflammatory infiltrate was seen in a case. Canine pseudo-placentational endometrial hyperplasia has very peculiar features, and it is a model for canine placentation and may help to better understand the cystic endometrial hyperplasia/pyometra complex.

Conservative treatments for feline fibroadenomatous changes of the mammary gland.

BACKGROUND: Fibroadenomatous changes (FAC) of the mammary gland in cats represents a benign, progesterone-associated fibroglandular proliferation of one or more mammary glands that may occur in intact female cats at the time of puberty, during pregnancy or pseudopregnancy, or in female and male cats of any age under progestin treatment. Nowadays, the elective treatment of FAC is based on the progesterone antagonist aglepristone. This study aimed to report the treatment of FAC with a combination of drugs designed to preserve mammary gland integrity, even in pregnant cats. CASE DESCRIPTION: Eight sexually intact female cats with FAC showed mammary glands that were symmetrically enlarged and inflamed in all cases, and the skin was ulcerated in six cats. Four cats were on days 25-32 of pregnancy at presentation. Non-pregnant cats were treated with aglepristone and with a dietary supplement containing maltodextrin and bromelain. The mammary glands were massaged daily with an Aloe vera emollient gel. If the gland was inflamed or ulcerated, broad-spectrum antimicrobial, and anti-inflammatory treatments were given, and the ulcers were treated topically with a hypericum and neem-based cream. Two of the four pregnant cats were treated with the same therapeutic schedule plus cloprostenol to facilitate uterine emptying. Two pregnant cats underwent the same protocol except for aglepristone, which was not administered to safeguard the litter and see if the adjuvant therapy worked independently. At term, they delivered four and three kittens, respectively, that were normally nursed and weaned after 40 days. In all the studied cases, the mammary gland reduced in size 2-3 weeks after the start of the treatment and completely remitted after 4-5 weeks. CONCLUSION: This case series encourages adjuvant therapy in the course of FAC to preserve mammary gland integrity and functionality. Exploiting the natural decline of progesterone at the end of pregnancy (or pseudopregnancy), the therapy may also be used without aglepristone, when its use has other limitations.

A case of neoplastic synchronism in a dog.

INTRODUCTION: Synchronous primary tumors are considered severe, comorbid conditions in people representing neoplasm that develop independently and concomitantly. A diagnosis of synchronous tumors was made in a dog, underlying the difficulties to reach it without the aid of multiple diagnostic techniques aimed to demonstrate the simultaneous coexistence of different tumor types. MATERIALS AND METHODS: A 7-year-old male Boxer dog presented several tumors located on the skin of the left hind limb and the scrotal region. Moreover, additional tumors in the testicles, after palpation and ultrasound examination, were detected. Following diagnostic results, the cutaneous tumor, scrotum, and testes were surgically removed. RESULTS: Pathological investigations revealed the presence of five different tumors: a cutaneous mast cell tumor; a scrotal melanocytoma; three testicular neoplasms (Sertoli Sustentacular cell tumor, seminoma, and interstitial Leydig cell tumor). CONCLUSIONS: The present report describes a neoplastic synchronism due to the presence of five different primary tumors in a dog and, for the first time the presence of a collision testicular tumor together with other non-testicular primary tumors. The occasional finding underlines the importance of the knowledge of such conditions in the process of decision-making and in carrying out all the proper diagnostic procedures for a correct diagnosis and clinical staging.

Conjunctival Inverted Papilloma Progressing to Carcinoma. First Report in Horse.

A five-year-old, entire female Arabian horse with a 6-month history of a non-painful nodule on the conjunctiva of the right eye was evaluated. Ophthalmological examination showed a firm, smooth and fleshy conjunctival mass that raised the suspicion of a conjunctival neoplasm. Histological evaluations showed that the mass was composed of an endophytic growth consisting of numerous long papillary projections of hyperplastic stratified squamous epithelium supported by thin fibrovascular stalks. Typical features of squamous cell carcinoma with disorganized cell growth and infiltration of surrounding tissues were detectable within the mass. Inverted papilloma progressing to carcinoma was diagnosed. Follow-up examination showed that no local recurrence was present during the 12-month follow-up period. To the authors' knowledge, this is the first report describing the inverted papilloma in the horse and, due to its progression to squamous cell carcinoma, warns about the inclusion of the inverted papilloma in the differential diagnosis of conjunctival neoplasm and driven treatments.

Immune Cells and Immunoglobulin Expression in the Mammary Gland Tumors of Dog.

Inflammatory cells have a role in tumor progression and have prognostic and therapeutic potential. The immunohistochemical expression for Mast Cell Tryptase, Macrophage Marker, CD79a, IgA, IgM and IgG on 43 cases of canine mammary gland lesions was analyzed. In hyperplasia, a few B cells (BCs) and Tumor-Associated Macrophages (TAMs) were observed, while the number of Tumor-Associated Mast Cells (TAMCs) was the highest. In the peritumoral stroma of malignant lesions, low number of TAMCs and a high number of TAMAs and BCs were present. Immune cells of each type were always lower in the intratumoral than peritumoral stroma. Positivity to CD79a was also detected in the epithelial cells of simple and micropapillay carcinomas. Immunoglobulin reactivity was mainly located in the epithelial cells where an intense positivity to IgA and IgG and a weak positivity for IgM were detectable. On the basis of our preliminary results and literature data, we suggest that such cells and molecules could be directly involved in the biology of canine mammary gland tumors. In breast cancer, stromal inflammatory cells and cancer derived immunoglobulins have been correlated with the progression, malignancy and poor prognosis of the tumor. The results herein reported show that the dog's mammary gland epithelium also expresses immunoglobulins, and they mostly show a direct relationship with the infiltration of macrophages. In addition, this study shows that the infiltration of mast cells, B-cells and macrophages varies depending on the degree of malignancy of neoplasia.

Role of Erythropoietin in Cerebral Glioma: An Innovative Target in Neuro-Oncology.

BACKGROUND: Erythropoietin (EPO) is a cytokine primarily involved in the regulation of erythropoiesis. In response to hypoxia-ischemia, hypoxia-inducible factor 1 induces EPO production, which, in turn, inhibits apoptosis of erythroid progenitor cells. By the same mechanism and acting through other signaling pathways, EPO exerts neuroprotective effects. Increased resistance to hypoxia and decreased apoptosis are thought to be important mechanisms for tumor progression, including malignant glioma. Because recent studies have demonstrated that EPO and its receptor (EPOR) are expressed in several tumors and can promote tumor growth, in the present study, we investigated EPO and EPOR expression in human glioma and the effect of EPO administration in a rat model of glioma implantation. METHODS: Using Western blotting and immunohistochemical analysis, we examined the expression of EPO, EPOR, platelet endothelial cell adhesion molecule, and Ki-67 in human glioma specimens and experimentally induced glioma in rats. In the experimental setting, a daily dose of recombinant human EPO (rHuEPO) or saline solution were administered for 21 days in Fischer rats subjected to 9L cell line implantation. RESULTS: In both human and animal specimens, we found an increase in EPOR expression as long as the lesion presented with an increasing malignant pattern. A significant direct correlation was found between the expression of EPOR and Ki-67 and EPOR and platelet endothelial cell adhesion molecule in low- and high-grade gliomas. The rats treated with rHuEPO presented with significantly larger tumor spread compared with the saline-treated rats. CONCLUSIONS: The results of our study have shown that the EPO/EPOR complex might play a significant role in the aggressive behavior of high-grade gliomas. The larger tumor spread in rHuEPO-treated rats suggests a feasible role for EPO in the aggressiveness and progression of malignant glioma.

Cisplatin-induced peripheral neuropathy: neuroprotection by erythropoietin without affecting tumour growth.

This study examined the dose-dependent efficacy of erythropoietin (EPO) for preventing and/or treating cisplatin (CDDP) induced peripheral neurotoxicity (CINP), and its influence on tumour treatment and growth. Rats received eight intraperitoneal (ip) injections of 2 mg/kg CDDP twice weekly. EPO co-administered (50 or 10 microg/kg ip, three times/week) had a dose-dependent effect, partially preventing CINP, but 0.5 microg/kg ip was not effective. The neuroprotective effect lasted at least 5 weeks after the last dose of EPO and CDDP. In addition, EPO (50 microg/kg ip three times/week) after the last injection of CDDP still induced a significant recovery of CINP. In a separate experiment in rats bearing mammary carcinoma EPO treatment (50 microg/kg ip) given concurrently with CDDP (1.0 and 1.5 mg/kg twice a week for four weeks) was neuroprotective without influencing the effectiveness of the treatment or tumour growth. EPO thus appears to be an effective neuroprotectant that does not interfere with tumour treatment.

The role of erythropoietin in neuroprotection: therapeutic perspectives.

Nervous system diseases are very complex conditions comprising a large variety of local and systemic responses. Several therapeutic agents interfering with all or in part the biochemical steps that ultimately cause neuronal death have been demonstrated to be neuroprotective in preclinical models. However, all the agents so far investigated have inexorably failed in the phase III trials carried out. A large body of evidence suggests that the hormone erythropoietin (EPO), besides its well-known hematopoietic action, exerts beneficial effects in the central nervous system. EPO's effect has been assessed in several experimental models of brain and spinal cord injury thus becoming a serious candidate for neuroprotection. The use of EPO as neuroprotectant raises several questions. Besides dosage and therapeutic time window, the safety of recombinant EPO administration in the setting of nervous system diseases takes priority over all other questions. Although recombinant EPO seems to be potentially safe at the neuroprotective proved doses, cardiovascular or cerebrovascular events can occur as a result of its bone marrow stimulating activities. The successful trial using EPO in patients with ischemic stroke and the large body of experimental evidence encourages intensive evaluation of this cytokine to support safe and larger clinical trials in the near future.

Neuroprotection by erythropoietin administration after experimental traumatic brain injury.

A large body of evidence indicates that the hormone erythropoietin (EPO) exerts beneficial effects in the central nervous system (CNS). To date, EPO's effect has been assessed in several experimental models of brain and spinal cord injury. This study was conducted to validate whether treatment with recombinant human EPO (rHuEPO) would limit the extent of injury following experimental TBI. Experimental TBI was induced in rats by a cryogenic injury model. rHuEPO or placebo was injected intraperitoneally immediately after the injury and then every 8 h until 2 or 14 days. Forty-eight hours after injury brain water content, an indicator of brain edema, was measured with the wet-dry method and blood-brain barrier (BBB) breakdown was evaluated by assay of Evans blue extravasation. Furthermore, extent of cerebral damage was assessed. Administration of rHuEPO markedly improved recovery from motor dysfunction compared with placebo group (P<0.05). Brain edema was significantly reduced in the cortex of the EPO-treated group relative to that in the placebo-treated group (80.6+/-0.3% versus 91.8%+/-0.8% respectively, P<0.05). BBB breakdown was significantly lower in EPO-treated group than in the placebo-treated group (66.2+/-18.7 mug/g versus 181.3+/-21 mug/g, respectively, P<0.05). EPO treatment reduced injury volume significantly compared with placebo group (17.4+/-5.4 mm3 versus 37.1+/-5.3 mm3, P<0.05). EPO, administered in its recombinant form, affords significant neuroprotection in experimental TBI model and may hold promise for future clinical applications.