Whole Mitochondrial Genome Sequencing Analysis of Canine Testicular Tumours.

Currently, the molecular background based on mitochondrial DNA (mtDNA) analysis of canine testicular tumours is underestimated. The available data mostly focus on histopathological evaluations, with a few reports of nuclear genome (nDNA) studies. Tumourigenesis represents a highly complex and diverse genetic disorder, which can also encompass defects in mtDNA. The aim of this study was to identify molecular changes in whole mitochondrial genome sequences obtained from dogs affected by testicular tumours. Samples of blood, tumour, and healthy tissue were collected from each animal, and mtDNA (ultimately 45 samples) was subsequently sequenced. Thereafter, protein analyses were performed to assess the impact of the identified molecular alterations on the amino acid level. The total number of observed changes included 722 SNPs, 12 mutations, 62 indels, 5 indel mutations, and 35 heteroplasmic sites. The highest number of mtDNA variants in protein-coding genes COX1, COX3, ATP6, ND1, ND4, and ND5 was observed. Interestingly, SNPs were found in 10 out of 22 tRNA genes. Most of the identified mtDNA defects were synonymous changes at the amino acid level. Also, polymorphisms and heteroplasmy were frequently observed in the variable number of tandem repeat (VNTR) regions, especially in its fragment spanning 16,138-16,358 bp. Based on the obtained results, it was possible to select 11 polymorphisms that occurred in all the tested samples (benign, malignant) and an additional five SNPs identified only in benign neoplasms. The comprehensive analysis of malignant testicular tumours demonstrated a significant diversity in their molecular profiles, with changes ranging from 17 to 101 per sample.

The diagnostic value of selected immune parameters in peripheral blood of dogs with malignant mammary tumours - a preliminary study.

Introduction: The main adaptive immune cells are T and B lymphocytes and they play key roles in the induction of immune responses against canine mammary tumours. Investigating these cell subpopulations may lead to more precise diagnosis of these malignancies. Material and Methods: The percentages of CD3+, CD4+ and CD8+ T cells and of CD21+ B cells in the peripheral blood of bitches with malignant mammary tumours were compared with those in the blood of healthy animals. The phenotypic features of peripheral blood leukocytes were evaluated by flow cytometry. Results: There was a significant difference in the mean percentages of CD3+ lymphocytes between healthy (66.7%) and metastatic dogs (46.1%), and between tumour-bearing non-metastatic (66.6%) and metastatic dogs. There was also a significant difference in CD4+ T helper cell percentages between healthy dogs (40.4%) and dogs with metastases (23.2%), and between the latter and dogs without them (35.5%). In the case of CD21+ lymphocyte subsets, a significant difference was noted between healthy animals (10.9%) and those with metastases (20.1%), and between the latter and patients without metastases (8.5%). There were also significant differences in CD3+/CD21+ ratios between the group with metastases (3.0), the healthy group (7.8), and the group without metastases (8.5). Similarly, a significant difference was noted in CD4+/CD8+ ratios between animals with metastases (1.4), bitches in the control group (2.2), and dogs without metastases (1.9). Conclusion: Peripheral blood leukocyte phenotypic characteristics are putative novel biomarkers. These findings may be useful in future studies improving mammary tumour diagnostic procedures, especially in metastasis detection.

Ehlers-Danlos syndrome (cutaneous asthenia) in a Campbell's dwarf hamster (Phodopus campbelli).

A case of cutaneous asthenia in a Campbell's dwarf hamster is described. The animal was found to have hyperextensible skin, glaucoma and lens dislocation. Histopathological examination revealed an irregular, haphazard arrangement of collagen fibres in the dermis. The animal underwent surgical reduction of the skin folds which provided only temporary relief.

Un cas d'asthénie cutanée chez un hamster nain de Campbell est décrit. L'animal présente une peau hyperextensible, un glaucome et une luxation du cristallin. L'examen histopathologique révèle une disposition irrégulière et désordonnée des fibres de collagène dans le derme. L'animal a subi une réduction chirurgicale des plis cutanés qui n'a apporté qu'un soulagement temporaire.

Descreve­se um caso de astenia cutânea em um hamster anão de Campbell. O animal apresentou hiperextensibilidade cutânea, glaucoma e deslocamento de lentes. Ao exame histopatológico, observou­se um arranjo irregular e aleatório das fibras colágenas na derme. O animal foi submetido à redução cirúrgica das dobras cutâneas que resultou em alívio apenas temporário.

Se describe un caso de astenia cutánea en un hámster enano Campbell. Se examinó al animal obsevando piel hiperextensible, glaucoma y luxación del cristalino. El examen histopatológico reveló una orientación aleatoria e irregular de las fibras de colágeno en la dermis. Se utilizó una técnica de reducción quirúrgica de los pliegues de la piel que solo indujo una mejora temporal.

Using Super-Resolution for Enhancing Visual Perception and Segmentation Performance in Veterinary Cytology.

The primary objective of this research was to enhance the quality of semantic segmentation in cytology images by incorporating super-resolution (SR) architectures. An additional contribution was the development of a novel dataset aimed at improving imaging quality in the presence of inaccurate focus. Our experimental results demonstrate that the integration of SR techniques into the segmentation pipeline can lead to a significant improvement of up to 25% in the mean average precision (mAP) metric. These findings suggest that leveraging SR architectures holds great promise for advancing the state-of-the-art in cytology image analysis.

Occurrence of mammary gland tumours in male dogs and its weak association with development of testicular tumours: a review.

Mammary gland tumours (MGTs) are commonly occurring neoplasms in female dogs. However, rare cases of MGTs in male dogs have been reported for years. Due to the low incidence of MGTs in male dogs in comparison to female dogs, veterinary oncology is mainly focused on mammary neoplasms diagnosed in female dogs and extensive research is conducted in this scientific area. Therefore, there are no sufficient epidemiological data on male dogs and the aetiology of their tumour development is still poorly understood.The aim of this literature review was to present cases of MGTs in male dogs for better understanding the scale of the problem over the years. The analyses of 74 affected male dogs with 92 tumours showed that the majority of MGTs in male dogs were benign tumours (54.3%), especially in form of adenomas, often developed in posterior canine mammary glands (58.1%).The increased number of canine MGTs in male dogs aged 7 -13 years with an age peak at 11 years was noted. The age of affected animals was not related to breed. Mammary gland neoplasms were diagnosed predominately in Crossbreeds (20.2%) followed by Cocker Spaniels (18.9%) and German Shepherds (10.8%).The association between MGT development in male dogs and co-occurrence of testicular tumours (TTs) has been discussed for years. Thus, cases of development of both tumours were included in this study. As a result, only in 12.7% cases of MGTs also history of TTs was described. Therefore, no general association between these tumours should be assumed.

Immunoreactivity of p21, MMP-1 and CB2 receptor proteins in cutaneous canine mast cell tumours: an association with the three-tier grading system.

Introduction: Mast cell tumours (MCTs) arise in the dermis and subcutaneous tissues in animals and humans and are one of the most common neoplasms of the skin in dogs. Cannabinoid type 2 receptor (CB2R), cyclin-dependent kinase inhibitor (p21) and matrix metalloproteinase 1 (MMP-1) are potential targets for novel anti-tumour therapeutic strategies. This study evaluated by immunohistochemical means the reactivity of p21, MMP-1 and CB2R proteins in association with a three-tier grading system in cutaneous canine MCTs. Material and Methods: Formalin-fixed, paraffin-embedded canine MCTs were processed for histochemical analysis and immunohistochemical staining using antibodies against p21, MMP-1 and CB2R. The results were analysed statistically. Results: The strongest p21 immunolabelling was detected in grade 3 MCTs, while grade 1 tumours showed mild or no detectable p21 immunoreactivity (P-value < 0.001). Strong immunolabelling of MMP-1 was the most common in grade 1 tumours (P-value < 0.001) and CB2R was significantly less frequent in grade 3 tumours than in grade 1 (P-value < 0.001) and grade 2 (P-value < 0.001). Conclusion: High immunoreactivity of MMP-1 can be a marker of grade 1 MCTs in dogs, whereas p21 protein overexpression can be a marker of grade 3 canine MCTs. Strong CB2R immunoreactivity with simultaneous underexpression of p21 and high immunoreactivity of MMP-1 proteins may indicate that the use of cannabinoids in grade 1 MCTs in dogs is practicable.

Incidence and the risk of occurrence of benign and malignant canine skin tumours in Poland - a five-year retrospective study.

Introduction: The aim of the study was to compile data on the frequency and distribution of canine skin tumours and determine the risk of these being malignant as opposed to benign. This determination proceeded from tumour histogenesis and gave consideration to the dog's breed, sex, age and the anatomical location of tumours. Material and Methods: This retrospective five-year epidemiological study included 3,139 canine skin tumours collected in Poland. A univariable logistic regression analysis was performed to determine the odds ratios (ORs) with 95% confidence intervals (CIs). Results: Microscopic analysis showed a significant predominance of benign tumours (65.02%) as well as mesenchymal and melanocytic tumours (59.57%). The most frequently diagnosed were mast cell tumours, accounting for 13.79% of all skin tumours, and other common tumour types were lipomas (6.40%), haemangiopericytomas (5.96%) and malignant melanomas (4.65%). The risk of malignant versus benign tumours was 1.212 times higher in the female than in the male dogs. A higher risk of development of malignant epithelial tumours was found in boxers (OR 4.091), German shepherds (OR 4.085) and flat-coated retrievers (OR 43.596). A higher risk of development of malignant mesenchymal tumours was found in golden retrievers (OR 4.693), boxers (OR 2.342), bulldogs (OR 3.469) and Maltese (OR 2.757). Conclusion: The results may serve as a reference point for further studies of the complex biology of canine skin tumours.

Topoisomerase IIα immunoexpression in feline mammary carcinomas: A correlation with Ki67 immunoexpression and the mitotic count.

The aim of the study was to compare the immunohistochemical expression of topoisomerase IIα protein (Topo IIα) with Ki67 expression and mitotic count in feline mammary carcinomas (FMCs). Topo IIα is considered as a proliferation indicator as well as a molecular target of anthracycline chemotherapy. The studied material included 70 FMCs from female cats treated with mastectomy. Primary mouse monoclonal antibodies directed against Topo IIα and Ki67 were used in immunohistochemical reactions. The number of mitotic figures was counted at 400× magnification in a field of 2.37 mm2. Immunohistochemical reaction for Topo IIα occurred in cell nuclei. The Topo IIα index ranged from 6.12% to 54.60% and was positively correlated with the values of the Ki67 index (r = 0.7193) and the mitotic count (r = 0. 2858). This indicates the potential possibility of use of the immunohistochemical expression of Topo IIα to assess the rate of proliferation in FMCs. The wide range of expression of Topo IIα in individual tumorus found in the conducted studies allows us to hypothesize that its assessment could be used as a predictive marker in chemotherapy of FMCs with the use of anthracyclines. However, this requires confirmation in clinical trials.

The use of optical coherence tomography for skin evaluation in healthy rats.

BACKGROUND: Noninvasive diagnostic techniques allow for morphological and morphometric in vivo evaluation of the skin. Optical coherence tomography (OCT) is a method that allows visualization of dermal structures up to several 100 µm with a resolution of 3-7.5 µm. OBJECTIVES: The aim of the study was the morphological and morphometric assessment of rats' skin using SD-OCT. ANIMALS: Fifteen male Wistar rats, aged 3 and 8 months, weighing 350-450 g. MATERIALS AND METHODS: The skin of the plantar metatarsal area of the right pelvic limb was assessed. The measurements were performed using Spectral Domain Optical Coherence Tomography (SD-OCT) scans and histological images. RESULTS: Spectral Domain Optical Coherence Tomography could determine the boundary between the epidermis and the dermis. Apart from the stratum corneum (SC), it did not allow for the differentiation of the individual epidermal layers. In the SD-OCT and in the histological examination, the mean thicknesses of the layers (µm ± SD) were (respectively): SC 33.053 ± µm(SD 5.85, 29.675 ± 5.54; epidermis 88.2 ± 7.97, 65.126 ± 13.23; dermis 259.86 ± 18.29, 166.05 ± 31.88 µm. There was a correlation between the total epidermal SD-OCT and histological measurements (r = 0.43, p = 0.05). Bland-Altman plots revealed a bias of -19.18 (95% confidence interval) -39.21 to 0.84 µm) in the case of live epidermis (stratum granulosum, stratum spinous, stratum basale). CONCLUSIONS AND CLINICAL RELEVANCE: Spectral Domain Optical Coherence Tomography can be used to evaluate rat epidermis and dermis. The method enables the differentiation of the SC, as well as the epidermis and the dermis. SD-OCT and histological thickness dimensions of the epidermis and skin differ.

Contexte - Les techniques de diagnostic non invasives permettent une évaluation morphologique et morphométrique in vivo de la peau. La tomographie par cohérence optique (OCT) est une méthode qui permet de visualiser les structures dermiques jusqu'à plusieurs centaines de micromètres avec une résolution de 3 à 7,5 µm. Objectifs - Le but de l'étude était l'évaluation morphologique et morphométrique de la peau des rats par SD-OCT. Animaux - Quinze rats Wistar mâles, âgés de 3 et 8 mois, pesant 350-450 g. Matériels et méthodes - La peau de la zone métatarsienne plantaire du membre pelvien droit a été évaluée. Les mesures ont été réalisées à l'aide de scaners SD-OCT (Spectral Domain Optical Coherence Tomography) et d'images histologiques Résultats - Le SD-OCT a pu déterminer la frontière entre l'épiderme et le derme. En dehors de la couche cornée (SC), il n'a pas permis la différenciation des couches épidermiques individuelles. Au SD-OCT et à l'examen histologique, les épaisseurs moyennes des couches (µm ± SD) étaient (respectivement) : SC 33,053 ± µm (SD 5,85, 29,675 ± 5,54 ; épiderme 88,2 ± 7,97, 65,126 ± 13,23 ; derme 259,86 ± 18,29, 166,05 ± 31,88 µm). Il y avait une corrélation entre le SD-OCT épidermique total et les mesures histologiques (r = 0,43, P = 0,05) Les tracés de Bland-Altman ont révélé un biais de -19,18 (intervalle de confiance à 95 %) -39,21 à 0,84 µm) dans le cas d'épiderme vivant (stratum granulosum, stratum spinous, stratum basale). Conclusions et pertinence clinique - Le SD-OCT peut être utilisé pour évaluer l'épiderme et le derme de rat. La méthode permet de différencier le SC, ainsi que l'épiderme et le derme. Le SD-OCT et les dimensions d'épaisseur histologiques de l'épiderme et de la peau diffèrent.

Introducción - las técnicas de diagnóstico no invasivas permiten la evaluación morfológica y morfométrica in vivo de la piel. La tomografía de coherencia óptica (OCT) es un método que permite la visualización de estructuras dérmicas de hasta varios cientos de micrómetros con una resolución de 3 a 7,5 µm. Objetivos - El objetivo del estudio fue la evaluación morfológica y morfométrica de la piel de ratas mediante SD-OCT. Animales - Quince ratas Wistar macho, de 3 y 8 meses de edad, con un peso de 350 a 450 g. Materiales y métodos - Se evaluó la piel de la zona metatarsiana plantar del miembro pélvico derecho. Las mediciones se realizaron utilizando escaneos SD-OCT (Tomografía de Coherencia Óptica de Dominio Espectral) e imágenes histológicas. Resultados - SD-OCT pudo determinar el límite entre la epidermis y la dermis. Aparte del estrato córneo (SC), no permitía la diferenciación de las capas epidérmicas individuales. En la SD-OCT y en el examen histológico, los espesores medios de las capas (µm ± SD) fueron (respectivamente): SC 33.053 ± µm (SD 5.85, 29.675 ± 5.54; epidermis 88.2 ± 7.97, 65.126 ± 13.23; dermis 259.86 ± 18,29, 166,05 ± 31,88 µm. Hubo una correlación entre el SD-OCT epidérmico total y las mediciones histológicas (r = 0,43, P = 0,05). Las gráficas de Bland-Altman revelaron un sesgo de -19,18 (intervalo de confianza del 95 %) -39,21 a 0,84 µm) en el caso de epidermis viva (estrato granuloso, estrato espinoso, estrato basal). Conclusiones y relevancia clínica- SD-OCT se puede utilizar para evaluar la epidermis y la dermis de rata. El método permite diferenciar el SC, así como la epidermis y la dermis. Las dimensiones de grosor de la epidermis y la piel difieren entre SD-OCT y la histología.

Contexto - Técnicas diagnósticas não invasivas permitem a avaliação morfológica e morfométrica in vivo da pele. A tomografia de coerência óptica (OCT) é um método que possibilita a visualização de estruturas dérmicas de até centenas de micrômetros com uma resolução de 3-7,5 µm. Objetivos - O objetivo do estudo foi a avaliação morfométrica e morfológica da pele de ratos utilizando SD-OCT. Animais - Quinze ratos Wistar machos, de três a oito meses de idade, pesando de 350 a 450 g. Materiais e métodos - A pele da região metatársica plantar do membro pélvico direito foi avaliada. As mensurações foram realizadas utilizando o SD-OCT (Spectral Domain Optical Coherence Tomography) e imagens histológicas. Resultados - A SD-OCT foi capaz de determinar o limite entre derme e epiderme. A exceção do estrato córneo (SC), não foi possível a diferenciação das camadas da epiderme individualmente. Na SD-OCT e na avaliação histológica, as espessuras médias das camadas fora, (µm ± DP), respectivamente: SC 33,053 ± 5,85; 29,675 ± 5,54; epiderme 88,2 ± 7,97; 65,126 ± 13,23; derme 259,86 ± 18,29; 166,05 ± 31,88 µm. Houve correlação entre SD-OCT e avaliação histológica nas mensurações da epiderme total (r = 0,43, P = 0,05). Os gráficos de Bland-Altman revelaram um viés de -19,8% (95% de intervalo de confiança) - 39,21 a 0,84 µm) no caso da epiderme viva (estrato espinhoso, estrato granuloso e estrato basal). Conclusões e relevância clínica - SD-OCT pode ser utilizada para avaliar a derme e a epiderme de ratos. Este método permite a diferenciação do SC, bem como da derme e epiderme. Pode haver discrepância na espessura da epiderme e da pele na SD-OCT e análise histológica.

Comparative Expression Analysis of Innate Immune Markers and Phagocytic Activity in Peripheral Blood of Dogs with Mammary Tumors.

Canine innate immune system role in cancer prevention and progression remains poorly understood. It has been revealed that innate immune cells could play a dual role in cancer immunology promoting or inhibiting tumor development and growth. Current immunotherapies target mainly the adaptive anti-tumor response and that may be a reason why they remain ineffective in a majority of patients. It is important to acquire detailed knowledge about innate immune mechanisms to broaden the diagnostic and therapeutic options and employ innate immune cells in anti-cancer therapies. In the present study, 21 female dogs of different breeds and types of spontaneous mammary tumors were investigated. The study aimed to find simple and cheap markers that can be used for preliminary diagnosis, prior to the surgical resection of the tumor. The differences in innate immune cell quantity and function were investigated between female dogs with malignant mammary tumors of epithelial and mesenchymal origin. Flow cytometry was used to evaluate the percentages of CD5+ lymphocytes including CD5low lymphocytes, CD11b integrin expression on leukocytes, phagocytosis, and oxidative burst. The number of CD11b lymphocytes was increased in tumors with epithelial origin compared to the control group. No significant differences were found between the percentages of phagocytic cells neither for granulocytes nor for monocytes. However, the phagocytes of canine patients with tumors of epithelial origin showed increased phagocytosis compared to the control group. The percentages of granulocytes that produced reactive oxygen species (ROS) in response to E.coli and PMA were not altered in patients with malignant tumors compared to control. A statistically significant difference between the number of ROS produced by the single granulocyte was demonstrated only between the group of bitches with epithelial tumors and the control group in case of E. coli stimulation. The obtained results suggest that some innate immune cells may be involved in anti-tumor immune mechanisms and have the potential to be supportive diagnostic markers in canine mammary tumors.