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OBJECTIVE: To evaluate the use of laser Doppler flowmetry and spectrophotometry (LDFS) for large intestinal viability assessment in horses with naturally occurring large intestinal strangulations. METHODS: By use of LDFS, intestinal microperfusion was quantified as tissue oxygen saturation (tSo2), hemoglobin (tHB), and blood flow (tBF) in cases with large colon volvulus and small colon strangulations undergoing colic surgery (n = 17). Intestinal biopsies were taken from the pelvic flexure in all large colon cases and in small colon cases that underwent intraoperative euthanasia. Measurements were compared between survivors and nonsurvivors, and the correlation between LDFS and (immuno)histology was tested (P < .05). RESULTS: The tSo2 and tBF were clearly lower and tHB was higher than previously reported in healthy horses. Following correction of the lesion, pelvic flexure tBF was significantly lower than that of the left ventral colon. Prior to correction of the lesion, microperfusion did not differ between survivors and nonsurvivors, but following release of the strangulation the survivors had a significantly higher tSo2 and tBF compared to the nonsurvivors. There was a negative correlation between tBF and interstitium-to-crypt ratio and a positive correlation between tHB and the histological hemorrhage score. There were no significant correlations between LDFS measurements and inflammatory cell counts or hypoxia-inducible factor-1α immunoreactivity. CONCLUSIONS: Large intestinal microperfusion was decreased in nonsurvivors compared to survivors and was correlated with histological injury, suggesting that LDFS has the potential to predict tissue injury and postoperative survival. CLINICAL RELEVANCE: The use of LDFS as an ancillary diagnostic aid may improve intraoperative viability assessment during colic surgery.
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An adult, 362 g, male, intact inland bearded dragon (Pogona vitticeps) was admitted to a veterinary clinic due to a temporary cloacal prolapse and a two-week history of reduced overall condition and forage intake. Physical examination revealed an approximately 2 × 1 cm round-shaped, rigid intracoelomic tissue mass. Multiple sand deposits were present on the cloacal mucous membranes, though no signs of cloacal prolapse were present. The lizard was otherwise responsive but showed reduced body tension and movement behavior. Initial fecal examination revealed a high-grade oxyuriasis. A 2 × 1.5 cm sized intracoelomic, well-vascularized, round-shaped mass was subsequently visualized by ultrasonography. After a two-day stabilization therapy, the intracoelomic mass was removed by performing a standard ventral coeliotomy under general anesthesia. Histopathological examination of the excised mass revealed an acinar pancreatic adenocarcinoma with infiltration of the peritumorous connective soft tissue. The lizard remained at the clinic for a further seven days. Its postsurgical condition improved slowly. However, the lizard started regular forage intake 10 days after surgery, and general behavior enhanced constantly within the following three weeks. The animal was presented for a follow-up six weeks after surgery, showing bright and alert behavior with no signs of disease or illness. The lizard was re-examined 20 months after the initial presentation due to a reduced overall condition and reduced food intake. Blood chemistry evaluation revealed markedly decreased protein parameters, and moderate ascites was identified ultrasonographically. A distinct association with the preceding neoplastic disease could not be made, and the lizard returned to its regular condition under supportive therapy within three weeks. To the authors' knowledge, this is the first report of successful treatment of a pancreatic carcinoma in a bearded dragon.
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Influenza A virus (IAV) infections in swine are usually subclinical, but they can reach high morbidity rates. The mortality rate is normally low. In this study, six vaccinated, spontaneously deceased sows revealed IAV infection and enhanced neutrophilic bronchopneumonia with unexpectedly large numbers of infiltrating eosinophils. The purpose of this study was to characterize these lung lesions with special emphasis on the phenotypes of inflammatory cells, the presence of eosinophilic peroxidase (EPO), and neutrophil extracellular traps (NETs). The number of Sirius red-stained eosinophils was significantly higher in the lungs of IAV-infected sows compared to healthy pigs, indicating a migration of eosinophils from blood vessels into the lung tissue stimulated by IAV infection. The detection of intra- and extracellular EPO in the lungs suggests its contribution to pulmonary damage. The presence of CD3+ T lymphocytes, CD20+ B lymphocytes, and Iba-1+ macrophages indicates the involvement of cell-mediated immune responses in disease progression. Furthermore, high numbers of myeloperoxidase-positive cells were detected. However, DNA-histone-1 complexes were reduced in IAV-infected sows, leading to the hypothesis that NETs are not formed in the IAV-infected sows. In conclusion, our findings in the lungs of IAV-infected vaccinated sows suggest the presence of so far unreported field cases of vaccine-associated enhanced respiratory disease.
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Vírus da Influenza A , Vacinas contra Influenza , Pulmão , Infecções por Orthomyxoviridae , Doenças dos Suínos , Animais , Suínos , Pulmão/patologia , Pulmão/virologia , Pulmão/imunologia , Doenças dos Suínos/virologia , Doenças dos Suínos/imunologia , Infecções por Orthomyxoviridae/imunologia , Infecções por Orthomyxoviridae/veterinária , Feminino , Vacinas contra Influenza/imunologia , Vacinas contra Influenza/administração & dosagem , Vírus da Influenza A/imunologia , Surtos de Doenças/veterinária , Eosinófilos/imunologia , Armadilhas Extracelulares/imunologia , Vacinação/veterinária , Peroxidase de Eosinófilo/metabolismoRESUMO
BACKGROUND: Ancillary diagnostic methods to enhance the accuracy of viability assessment have not been established for use in clinical practice. OBJECTIVES: To assess intestinal microperfusion measured by Laser Doppler Flowmetry and Spectrophotometry (LDFS) in naturally occurring small intestinal strangulations of different origins and to compare this between viable and non-viable segments. STUDY DESIGN: Prospective clinical trial. METHODS: Forty horses undergoing colic surgery for naturally occurring small intestinal strangulations were included. Tissue oxygen saturation (tSO2), haemoglobin (tHB) and blood flow (tBF) were determined by LDFS before and after release of the strangulation. Intestinal biopsies were taken in cases that underwent intestinal resection or intraoperative euthanasia and assessed using a semi-quantitative mucosal injury score (MIS). The LDFS measurements were compared between the different categories of strangulation causes and histopathological injury using parametric and non-parametric tests (p < 0.05). RESULTS: Strangulations by pedunculated lipomas had lower tBF (13.9 ± 18 arbitrary units [AU]) than epiploic foramen entrapments (65.2 ± 61 AU; CI -1.697 to -0.2498; p = 0.005). Segments with MIS > 5 showed lower tBF during strangulation than segments with MIS < 4 (mean difference 61.1 AU; CI -1.119 to -0.07361; p = 0.03). This did not differ significantly following release of strangulation. Furthermore, there was a positive correlation between the inflammatory cell count and tBF during strangulation (r 0.34; CI 0.01 to 0.60; p = 0.04). The tSO2 and tHB did not differ between the different categories of lesions or injury. MAIN LIMITATIONS: No biopsies could be taken from the intestinal segments that did not undergo resection. The duration of strangulation could not reliably be ascertained. CONCLUSIONS: Blood flow measurements in naturally occurring strangulating lesions show a varying degree of ischaemia in different causes of strangulation. Intestinal blood flow measurements prior to release of the strangulation could potentially contribute to the identification of mucosal injury, yet a high individual variability and other contributing factors need to be considered.
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OBJECTIVE: To assess the histological injury and intestinal microperfusion measured by laser Doppler flowmetry and spectrophotometry (LDFS) of the small intestine orad to a strangulation during colic surgery. ANIMALS: Horses with naturally occurring small intestinal strangulations undergoing colic surgery were included. METHODS: In this prospective clinical trial, intestinal tissue oxygen saturation (tSO2) and tissue blood flow (tBF) were measured by LDFS orad to the strangulation following release of the strangulation (n = 18). The number of horses with postoperative reflux (POR) and the cases that survived until discharge were compared between groups using Fisher's exact test (P < .05). Intestinal biopsies were taken in cases that underwent intestinal resection or intraoperative euthanasia (n = 28). Measurements were compared between injured and noninjured segments with a Mann-Whitney U or t test. RESULTS: The tSO2 and tBF of the orad intestine were lower than previously reported in healthy horses. Horses with low tSO2 of < 35% were significantly more likely to suffer from POR (6/6 cases) compared to cases with tSO2 > 69% (1/6). The number of horses that survived were not statistically different between these groups (2/6 and 6/6). All horses with mucosal injury developed POR (6/6), which was significantly more likely compared to horses without mucosal injury (3/13). No significant difference in tSO2 or tBF could be found between the segments with and without histological injury. CLINICAL RELEVANCE: The results suggest that measuring tSO2 in the orad segment during colic surgery may aid in predicting postoperative issues.
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Cólica , Doenças dos Cavalos , Complicações Pós-Operatórias , Animais , Cavalos , Doenças dos Cavalos/cirurgia , Doenças dos Cavalos/metabolismo , Cólica/veterinária , Cólica/cirurgia , Complicações Pós-Operatórias/veterinária , Feminino , Masculino , Estudos Prospectivos , Íleus/veterinária , Saturação de Oxigênio , Fluxometria por Laser-Doppler/veterinária , Oxigênio/metabolismo , Intestino Delgado/cirurgia , Espectrofotometria/veterináriaRESUMO
An adult male Bell's hinge-back tortoise (Kinixys belliana) was admitted to a veterinary clinic due to a swelling in the oral cavity. Physical examination revealed an approximately 2.5 × 1.5 cm sized, irregularly shaped tissue mass with villiform projections extending from its surface located in the oropharyngeal cavity. An initial biopsy was performed, and the lesion was diagnosed as squamous papilloma. Swabs taken for virological examination tested negative with specific PCRs for papillomavirus and herpesvirus. Further analysis of the oropharyngeal mass via metagenomic sequencing revealed sequence reads corresponding to a member of the family Adintoviridae. The tissue mass was removed one week after the initial examination. The oral cavity remained unsuspicious in follow-up examinations performed after one, five and twenty weeks. However, a regrowth of the tissue was determined 23 months after the initial presentation. The resampled biopsy tested negative for sequence reads of Adintoviridae. Conclusively, this report presents the diagnostic testing and therapy of an oral cavity lesion of unknown origin. The significance of concurrent metagenomic determination of adintovirus sequence reads within the tissue lesion is discussed.
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The presence of bronchus-associated lymphoid tissue (BALT) and its structural components has been described in different healthy animal species and in animals with diseases of the respiratory tract. In contrast to normal mammals, BALT is absent in healthy human adult lungs, but has been found in the lungs of children. The histological characteristics of organized mucosa-associated lymphoid tissue (MALT), its subsets of immune cells and their in situ distribution in the lung of healthy subadult and adult cattle shows close similarities with BALT in humans and other animal species such as sheep, horses and pigs. This study clearly demonstrates that organized MALT also occurs in the tracheal mucosa of cattle. The absence of tracheal MALT and BALT in calves suggest that these structures are not constitutive. In the mucosa of bovine trachea, bronchi and bronchioli, MHC II+ and CD11c+ dendritic cells (DCs) are located in the epithelium and in the lamina propria mucosae. These DCs are already present in calves soon after birth. Examination of tangential epithelial sheets shows that in the bovine tracheal epithelium, like in man and rat, a dense network of MHC II+ and CD11c+ DCs exists and that their number is considerably higher than in conventional transverse sections. In the bovine tracheal and bronchial epithelium, MHC II+ DCs are extending their dendrites towards the lumen indicating that these DCs possibly are involved in sampling of luminal antigens. The presence of significantly higher numbers of MHC II+ DCs in the tracheal and bronchial/bronchiolar mucosa of older cattle in than in calves possibly results from local stimulation with exogenous antigens during postnatal life. Detection of DCs expressing the costimulatory molecules CD80 and CD86 in calves and cattle suggests maturation of DCs, which is most likely induced by stimulation with exogenous antigens.
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Tecido Linfoide , Traqueia , Bovinos , Animais , Humanos , Ratos , Suínos , Cavalos , Ovinos , Pulmão , Brônquios , Células Apresentadoras de Antígenos , MamíferosRESUMO
Introduction: Hypoxia inducible factors (HIF) are widely researched in human medicine for their role in different disease processes. The aim of this study was to investigate the expression and distribution of HIF in experimental small intestinal ischemia in the horse. Methods: In 14 horses under general anesthesia, segmental jejunal ischemia with 90% reduction in blood flow was induced. The horses were randomly divided into two groups of seven horses, one subjected to ischemic postconditioning (IPoC) by delayed reperfusion, and a control group (group C) undergoing undelayed reperfusion. Intestinal samples were taken pre-ischemia, after ischemia and after reperfusion. Following immunohistochemical staining for HIF1α and -2α, the immunoreactivity pattern in the small intestine was evaluated by light microscopy, and the mucosal enterocyte and muscularis staining were semi-quantitatively scored. Additionally, mucosal HIF1α protein levels were determined by an Enzyme Linked Immunosorbent Assay (ELISA), and mRNA levels of HIF1α and its target genes by a two-step real-time Reverse Transcriptase Polymerase Chain Reaction. Statistical comparison was performed between the groups and time points using parametric and non-parametric tests (p < 0.05). Results: All cell types exhibited cytoplasmic and nuclear immunoreactivity for HIF1α. After reperfusion, the cytoplasmic staining of the crypt and villus enterocytes as well as the villus nuclear staining significantly increased, whereas the perinuclear granules in the crypts decreased. The protein levels showed a significant decrease in group C at reperfusion, with lower HIF1α levels in group C compared to group IPoC during ischemia and reperfusion. No other group differences could be detected. In the HIF2α stained slides, mild to moderate cytoplasmic staining yet no nuclear immunoreactivity of the enterocytes was observed, and no significant changes over time were noted. Discussion: the changes in HIF1α immunoreactivity pattern and expression over time suggest that this transcription factor plays a role in the intestinal response to ischemia in horses. However, the current study could not identify an effect of IPoC on HIF distribution or expression.
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Arachnoid cysts are cystic lesions that occur in spinal or intracranial locations in the leptomeningeal space. Four intracranial cases have been described in cats, three of which were diagnosed by imaging techniques alone. We now report the clinical, gross and histopathological findings in a 5-year-old, male-neutered European Shorthair cat that presented with chronic, asymmetrical encephalopathy. Using magnetic resonance imaging, a focal, fluid-filled cavity that did not show contrast enhancement was identified in the left temporal and piriform lobes. Necropsy confirmed the presence of a cystic, meningeal cavity filled with clear, serous fluid. Histologically, the cyst had an irregular, hypereosinophilic surface and single psammoma bodies with moderate perivascular oedema in the adjacent neuroparenchyma. Immunohistochemical evidence of meningeal tissue surrounding the cyst confirmed the diagnosis of an arachnoid cyst, which should be considered as a differential diagnosis of intracranial, fluid-filled cavities.
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Cistos Aracnóideos , Encefalopatias , Doenças do Gato , Animais , Gatos , Masculino , Cistos Aracnóideos/complicações , Cistos Aracnóideos/diagnóstico , Cistos Aracnóideos/veterinária , Doenças do Gato/diagnóstico por imagem , Imageamento por Ressonância Magnética/veterinária , Encefalopatias/diagnóstico por imagem , Encefalopatias/etiologia , Encefalopatias/veterináriaRESUMO
Mutations within the ectodysplasin A (EDA) gene have been associated with congenital hypotrichosis and anodontia (HAD/XHED) in humans, mice, dogs and cattle. We identified a three-generation family of Fleckvieh cattle with male calves exhibiting clinical and histopathological signs consistent with an X-linked recessive HAD (XHED). Whole genome and Sanger sequencing of cDNA showed a perfect association of the missense mutation g.85716041G>A (ss2019497443, rs1114816375) within the EDA gene with all three cases following an X-linked recessive inheritance, but normal EDAR and EDARADD. This mutation causes an exchange of glycine (G) with arginine (R) at amino acid position 227 (p.227G>R) in the second collagen triple helix repeat domain of EDA. The EDA variant was associated with a significant reduction and underdevelopment of hair follicles along with a reduced outgrowth of hairs, a complete loss of seromucous nasolabial and mucous tracheal and bronchial glands and a malformation of and reduction in number of teeth. Thermostability of EDA G227R was reduced, consistent with a relatively mild hair and tooth phenotype. However, incisors and canines were more severely affected in one of the calves, which correlated with the presence of a homozygous missense mutation of RNF111 (g.51306765T>G), a putative candidate gene possibly associated with tooth number in EDA-deficient Fleckvieh calves.
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Displasia Ectodérmica Anidrótica Tipo 1 , Displasia Ectodérmica , Hipotricose , Deformidades Congênitas dos Membros , Animais , Bovinos , Masculino , Camundongos , Displasia Ectodérmica/genética , Displasia Ectodérmica Anidrótica Tipo 1/genética , Mutação , Mutação de Sentido IncorretoRESUMO
The molecular heterogeneity of feline mammary carcinomas (FMCs) represents a prognostic and therapeutic challenge. RNA-Seq-based comparative transcriptomic profiling serves to identify recurrent and exclusive differentially expressed genes (DEGs) across sample types and molecular subtypes. Using mass-parallel RNA-Seq, we identified DEGs and performed comparative function-based analysis across 15 tumours (four basal-like triple-negative [TN], eight normal-like TN, and three luminal B fHER2 negative [LB fHER2-]), two cell lines (CL, TiHo-0906, and TiHo-1403) isolated from the primary tumours (LB fHER2-) of two cats included in this study, and 13 healthy mammary tissue controls. DEGs in tumours were predominantly upregulated; dysregulation of CLs transcriptome was more extensive, including mostly downregulated genes. Cell-cycle and metabolic-related DEGs were upregulated in both tumours and CLs, including therapeutically-targetable cell cycle regulators (e.g. CCNB1, CCNB2, CDK1, CDK4, GTSE1, MCM4, and MCM5), metabolic-related genes (e.g. FADS2 and SLC16A3), heat-shock proteins (e.g. HSPH1, HSP90B1, and HSPA5), genes controlling centrosome disjunction (e.g. RACGAP1 and NEK2), and collagen molecules (e.g. COL2A1). DEGs specifically upregulated in basal-like TN tumours were involved in antigen processing and presentation, in normal-like TN tumours encoded G protein-coupled receptors (GPCRs), and in LB fHER2- tumours were associated with lysosomes, phagosomes, and endosomes formation. Downregulated DEGs in CLs were associated with structural and signalling cell surface components. Hence, our results suggest that upregulation of genes enhancing proliferation and metabolism is a common feature among FMCs and derived CLs. In contrast, the dissimilarities observed in dysregulation of membrane components highlight CLs' disconnection with the tumour microenvironment. Furthermore, recurrent and exclusive DEGs associated with dysregulated pathways might be useful for the development of prognostically and therapeutically-relevant targeted panels.
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Carcinoma , Perfilação da Expressão Gênica , Animais , Biomarcadores , Gatos , Ciclo Celular/genética , Linhagem Celular , Proteínas de Choque Térmico/genética , Transcriptoma , Microambiente TumoralRESUMO
Tumors originating from eccrine glands are rare findings in dogs and cats. In most cases, the tumors are malignant, while adenomas are only reported anecdotally. In the present case, a one-year-old female, spayed cat was presented with a swelling of the footpad of the right forelimb. Initially, the mass possessed a diameter of 2 cm which progressed to 4 cm within the following two months. At the latter time point the tumor was ulcerated. After surgical removal, histological and immunohistochemical analyses were performed. Histologically, a well demarcated, nodular, multilobular mass was present. The cuboidal to columnar neoplastic cells were arranged in tubular and acinar structures. Tumor cells possessed large, round to oval nuclei with moderately distinct nucleoli. Mitotic figures averaged 0-1 per high power field. Additionally, large areas of chondroid metaplasia were evident. Immunohistochemically, neoplastic cells were positive for pan-cytokeratin AE1/AE3 whereas thyroid transcription factor 1 (TTF1) was not expressed. Based on the histological and immunohistochemical findings an adenoma of the eccrine glands was diagnosed.
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Adenoma , Doenças do Gato , Doenças do Cão , Adenoma/diagnóstico , Adenoma/cirurgia , Adenoma/veterinária , Animais , Doenças do Gato/diagnóstico , Doenças do Gato/cirurgia , Gatos , Doenças do Cão/diagnóstico , Doenças do Cão/cirurgia , Cães , FemininoRESUMO
Canine prostate cancer is classified into adenocarcinoma, transitional cell carcinoma with prostatic involvement, and mixed forms. Early metastatic spread leads to poor prognosis and limited treatment options. Masitinib is approved for the treatment of canine mast cell tumours and inhibits tyrosine kinase c-Kit, tyrosine-protein kinase Lyn (Lyn), and platelet-derived growth factor receptors alpha and beta (PDGFR-α, PDGFR-ß), which are known to be expressed in canine prostate cancer. The aim of this study was to evaluate masitinib in an in vitro model consisting of cell lines from primary prostate adenocarcinoma, the associated lymph node metastasis of the same patient, and transitional cell carcinoma. To assess the suitability of the model system, the targets of masitinib were investigated by immunocytochemistry in the cell lines and by immunohistochemistry in the respective formalin-fixed, paraffin-embedded (FFPE) original neoplastic tissue. After exposure to masitinib, cell viability, cell count, apoptosis induction, and protein expression of c-Kit, Lyn, PDGFR-α, and PDGFR-ß were assessed. To hedge the efficacy, two application protocols of masitinib (single application or 12-h double-dose regimen) were compared. Immunocytochemical and immunohistochemical analysis revealed increased Lyn, PDGFR-α, and PDGFR-ß expression in cell lines and FFPE original neoplastic tissue compared to healthy prostate tissue. Masitinib exposure increased apoptosis, while the cell counts and cell viability decreased in a dose- and application interval-dependent manner, with increased impact in the 12-h double-dose regimen. These in vitro effects of masitinib in canine prostate cancer and associated metastasis support further in vivo research and modifications of the clinical treatment protocol in future studies.
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Adenocarcinoma , Carcinoma de Células de Transição , Doenças do Cão , Neoplasias da Próstata , Adenocarcinoma/tratamento farmacológico , Adenocarcinoma/veterinária , Animais , Benzamidas , Carcinoma de Células de Transição/veterinária , Linhagem Celular , Doenças do Cão/tratamento farmacológico , Cães , Masculino , Piperidinas , Neoplasias da Próstata/tratamento farmacológico , Neoplasias da Próstata/veterinária , Proteínas Proto-Oncogênicas c-kit/metabolismo , Piridinas , Receptor alfa de Fator de Crescimento Derivado de Plaquetas , Receptor beta de Fator de Crescimento Derivado de Plaquetas , TiazóisRESUMO
Self-renewal of the intestinal epithelium originates from stem cells located at the crypt base. Upregulation of various stem cell markers in intestinal epithelial neoplasms indicates a potential role of stem cells in tumorigenesis. In this study, the immunoreactivity of potential intestinal stem cell markers (Sry box transcription factor 9 [Sox9], homeodomain-only protein [Hopx], survivin) and tuft cell marker doublecortin-like kinase 1 (DCLK1) in normal canine intestine and intestinal epithelial neoplasms was investigated. Formalin-fixed paraffin-embedded (FFPE) small and large intestine as well as intestinal neoplasms (55 colorectal adenomas [CRAs], 17 small intestinal adenocarcinomas [SICs], and 12 colorectal adenocarcinomas [CRCs]) were analyzed immunohistologically. Potential stem cell markers Sox9, Hopx, and survivin were detected in the crypts of normal canine small and large intestine. DCLK1+ tuft cells were present in decreasing numbers along the crypt-villus axis of the jejunum and rarely detectable in large intestine. In canine intestinal epithelial tumors, nuclear Sox9 immunoreactivity was detectable in 84.9% (CRA), 80% (CRC), and 77% of epithelial neoplastic cells (SIC). Hopx and survivin were expressed within cytoplasm and nuclei of neoplastic cells in benign and malignant tumors. DCLK1 showed a cytoplasmic reaction within neoplastic cells. The combined score of Hopx, DCLK1, and survivin varied among the examined cases. Overall, malignant tumors showed lower DCLK1 scores but higher Hopx scores in comparison with benign tumors. For survivin, no differences were detectable. In conclusion, stem cell markers Sox9, Hopx, and survivin were detectable at the crypt base and the immunoreactivity of Sox9, DCLK1, survivin, and Hopx was increased in canine intestinal adenomas and adenocarcinomas compared with normal mucosa.
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Adenocarcinoma , Adenoma , Neoplasias Colorretais , Doenças do Cão , Adenocarcinoma/patologia , Adenocarcinoma/veterinária , Adenoma/metabolismo , Adenoma/veterinária , Animais , Neoplasias Colorretais/metabolismo , Neoplasias Colorretais/veterinária , Doenças do Cão/metabolismo , Cães , Quinases Semelhantes a Duplacortina , Mucosa Intestinal/patologia , Intestinos/patologia , Proteínas Serina-Treonina Quinases , Survivina/metabolismoRESUMO
BACKGROUND: Canine prostate adenocarcinoma (PAC) and transitional cell carcinoma (TCC) are typically characterized by metastasis and chemoresistance. Cell lines are important model systems for developing new therapeutic strategies. However, as they adapt to culturing conditions and undergo clonal selection, they can diverge from the tissue from which they were originally derived. Therefore, a comprehensive characterization of cell lines and their original tissues is paramount. METHODS: This study compared the transcriptomes of nine canine cell lines derived from PAC, PAC metastasis and TCC to their respective original primary tumor or metastasis tissues. Special interests were laid on cell culture-related differences, epithelial to mesenchymal transition (EMT), the prostate and bladder cancer pathways, therapeutic targets in the PI3K-AKT signaling pathway and genes correlated with chemoresistance towards doxorubicin and carboplatin. RESULTS: Independent analyses for PAC, PAC metastasis and TCC revealed 1743, 3941 and 463 genes, respectively, differentially expressed in the cell lines relative to their original tissues (DEGs). While genes associated with tumor microenvironment were mostly downregulated in the cell lines, patient-specific EMT features were conserved. Furthermore, examination of the prostate and bladder cancer pathways revealed extensive concordance between cell lines and tissues. Interestingly, all cell lines preserved downstream PI3K-AKT signaling, but each featured a unique therapeutic target signature. Additionally, resistance towards doxorubicin was associated with G2/M cell cycle transition and cell membrane biosynthesis, while carboplatin resistance correlated with histone, m- and tRNA processing. CONCLUSION: Comparative whole-transcriptome profiling of cell lines and their original tissues identifies models with conserved therapeutic target expression. Moreover, it is useful for selecting suitable negative controls, i.e., cell lines lacking therapeutic target expression, increasing the transfer efficiency from in vitro to primary neoplasias for new therapeutic protocols. In summary, the dataset presented here constitutes a rich resource for canine prostate and bladder cancer research.
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BACKGROUND: Ischaemic postconditioning (IPoC) has been shown to ameliorate ischaemia reperfusion injury in different species and tissues. OBJECTIVES: To assess the feasibility of IPoC in equine small intestinal ischaemia and to assess its effect on histomorphology, electrophysiology and paracellular permeability. STUDY DESIGN: Randomised in vivo experiment. METHODS: Experimental jejunal ischaemia was induced for 90 min in horses under general anaesthesia. In the control group (C; n = 7), the jejunum was reperfused without further intervention. In the postconditioning group (IPoC; n = 7), reocclusion was implemented following release of ischaemia by clamping the mesenteric vessels in three cycles of 30 seconds. This was followed by 120 minutes of reperfusion in both groups. Intestinal microperfusion and oxygenation was measured during IPoC using spectrophotometry and Doppler flowmetry. Histomorphology and histomorphometry of the intestinal mucosa were assessed. Furthermore, electrophysiological variables and unidirectional flux rates of 3 H-mannitol were determined in Ussing chambers. Western blot analysis was performed to determine the tight junction protein levels of claudin-1, claudin-2 and occludin in the intestinal mucosa. Comparisons between the groups and time points were performed using a two-way repeated measures analysis of variance (ANOVA) or non-parametric statistical tests for the ordinal and not normally distributed data (significance P < .05). RESULTS: IPoC significantly reduced intestinal microperfusion during all clamping cycles yet affected oxygen saturation only during the first cycle. After reperfusion, Group IPoC showed significantly less mucosal villus denudation (mean difference 21.5%, P = .02) and decreased mucosal-to-serosal flux rates (mean difference 15.2 nM/cm2 /h, P = .007) compared to Group C. There were no significant differences between the groups for the other tested variables. MAIN LIMITATIONS: Small sample size, long-term effects were not investigated. CONCLUSIONS: Following IPoC, the intestinal mucosa demonstrated significantly less villus denudation and paracellular permeability compared to the untreated control group, possibly indicating a protective effect of IPoC on ischaemia reperfusion injury.
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Doenças dos Cavalos , Pós-Condicionamento Isquêmico , Traumatismo por Reperfusão , Animais , Doenças dos Cavalos/prevenção & controle , Cavalos , Intestino Delgado , Isquemia/veterinária , Pós-Condicionamento Isquêmico/veterinária , Jejuno , Traumatismo por Reperfusão/prevenção & controle , Traumatismo por Reperfusão/veterináriaRESUMO
We investigated a highly inbred family of British Shorthair cats in which two offspring were affected by deteriorating paraparesis due to complex skeletal malformations. Radiographs of both affected kittens revealed vertebral deformations with marked stenosis of the vertebral canal from T11 to L3. Additionally, compression of the spinal cord, cerebellar herniation, coprostasis and hypogangliosis were found. The pedigree suggested monogenic autosomal recessive inheritance of the trait. We sequenced the genome of an affected kitten and compared the data to 62 control genomes. This search yielded 55 private protein-changing variants of which only one was located in a likely functional candidate gene, LTBP3, encoding latent transforming growth factor ß binding protein 3. This variant, c.158delG or p.(Gly53Alafs*16), represents a 1 bp frameshift deletion predicted to truncate 95% of the open reading frame. LTBP3 is a known key regulator of transforming growth factor ß (TGF-ß) and is involved in bone morphogenesis and remodeling. Genotypes at the LTBP3:c.158delG variant perfectly co-segregated with the phenotype in the investigated family. The available experimental data together with current knowledge on LTBP3 variants and their functional impact in human patients and mice suggest LTBP3:c.158delG as a candidate causative variant for the observed skeletal malformations in British Shorthair cats. To the best of our knowledge, this study represents the first report of LTBP3-related complex skeletal dysplasia in domestic animals.
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Doenças do Gato/genética , Proteínas de Ligação a TGF-beta Latente/genética , Osteocondrodisplasias/veterinária , Animais , Doenças do Gato/diagnóstico por imagem , Gatos , Feminino , Mutação da Fase de Leitura , Genótipo , Endogamia , Masculino , Osteocondrodisplasias/diagnóstico por imagem , Osteocondrodisplasias/genética , Linhagem , Fenótipo , Radiografia/veterináriaRESUMO
Prostate cancer (PCa) in dogs is a highly malignant disease akin to its human counterpart. In contrast to the situation in humans, multi-gene approaches facilitating risk stratification of canine PCa are barely established. The aims of this study were the characterization of the transcriptional landscape of canine PCa and the identification of diagnostic, prognostic and/or therapeutic biomarkers through a multi-step screening approach. RNA-Sequencing of ten malignant tissues and fine-needle aspirations (FNA), and 14 nonmalignant tissues and FNAs was performed to find differentially expressed genes (DEGs) and deregulated pathways. The 4098 observed DEGs were involved in 49 pathways. These 49 pathways could be grouped into five superpathways summarizing the hallmarks of canine PCa: (i) inflammatory response and cytokines; (ii) regulation of the immune system and cell death; (iii) cell surface and PI3K signaling; (iv) cell cycle; and (v) phagosome and autophagy. Among the highly deregulated, moderately to strongly expressed DEGs that were members of one or more superpathways, 169 DEGs were listed in relevant databases and/or the literature and included members of the PCa pathway, oncogenes, prostate-specific genes, and druggable genes. These genes are novel and promising candidate diagnostic, prognostic and/or therapeutic canine PCa biomarkers.
Assuntos
Biomarcadores Tumorais/genética , Biologia Computacional/métodos , Regulação Neoplásica da Expressão Gênica , Redes Reguladoras de Genes , Neoplasias da Próstata/patologia , RNA-Seq/métodos , Transcriptoma , Animais , Cães , Perfilação da Expressão Gênica , Masculino , Neoplasias da Próstata/genéticaRESUMO
A 4-year-old, neutered male Husky-mix dog weighing 29.4â kg that reportedly ingested a mushroom most likely of the genus Amanita one day prior to presentation exhibited signs of diarrhea, vomitus, inappetence and progressively worsening lethargy. Clinical chemistry revealed hypoglycemia, hyperbilirubinemia, decreased prothrombin and thromboplastin time, as well as increased liver enzyme activities. Despite hospitalization and supportive therapy over a period of 3â days the dog's general condition worsened leading to euthanasia. The pathomorphological findings were characterized by hemorrhage in several organs, hemorrhagic ingesta, icterus, and marked hepatic cellular necrosis.
Assuntos
Doenças do Cão , Falência Hepática Aguda , Intoxicação Alimentar por Cogumelos , Amanita , Animais , Diarreia/veterinária , Doenças do Cão/diagnóstico , Doenças do Cão/etiologia , Cães , Falência Hepática Aguda/diagnóstico , Falência Hepática Aguda/etiologia , Falência Hepática Aguda/terapia , Falência Hepática Aguda/veterinária , Masculino , Intoxicação Alimentar por Cogumelos/complicações , Intoxicação Alimentar por Cogumelos/diagnóstico , Intoxicação Alimentar por Cogumelos/terapia , Intoxicação Alimentar por Cogumelos/veterináriaRESUMO
Campylobacter (C.) is the most common food-borne zoonosis in humans, which mainly manifests with watery to bloody diarrhoea. While C. jejuni is responsible for most cases of infection, C. coli is less frequently encountered. The object of the study was to prove the clinical impact of mono- and co-colonisation of C. coli and C. jejuni on weaned piglets in an infection model and to investigate the impact on transepithelial transport processes in the jejunum and caecum. At an age of eight weeks, eight pigs were infected with C. coli (ST-5777), 10 pigs with C. jejuni (ST-122), eight pigs with both strains, and 11 piglets served as control. During the four-week observation period, no clinical signs were observed. During dissection, both strains could be isolated from the jejunum and the caecum, but no alteration of the tissue could be determined histopathologically. Mono-infection with C. jejuni showed an impact on transepithelial ion transport processes of the caecum. An increase in the short circuit current (Isc) was observed in the Ussing chamber resulting from carbachol- and forskolin-mediated Cl- secretion. Therefore, we speculate that caecal colonisation of C. jejuni might affect the transport mechanisms of the intestinal mucosa without detectable inflammatory reaction.