Canine glioblastoma-derived extracellular vesicles as precise carriers for glioblastoma imaging: Targeting across the blood-brain barrier.

The treatment of glioblastoma (GBM) faces significant challenges due to the difficulty of delivering drugs through the blood-brain barrier (BBB). Extracellular vesicles (EVs) have emerged as potential carriers for targeted drug delivery to brain tumors. However, their use and distribution in the presence of an intact BBB and their ability to target GBM tissue are still under investigation. This study explored the use of EVs for GBM targeting across the BBB. Canine plasma EVs from healthy dogs and dogs with glioma were isolated, characterized, and loaded with diagnostic agents. Biodistribution studies were conducted in healthy murine models and a novel intranasal model that preserved BBB integrity while initiating early-stage GBM growth. This model assessed EVs' potential for delivering the contrast agent gadoteric acid to intracranial tumors. Imaging techniques, such as bioluminescence and MRI, confirmed EVs' targeting and delivery capabilities thus revealing a selective accumulation of canine glioma-derived EVs in brain tissue under physiological conditions. In the model of brain tumor, MRI experiments demonstrated the ability of EVs to accumulate gadoteric acid within GBM to enhance contrast of the tumoral mass, even when BBB integrity is maintained. This study underscores the potential of EVs derived from glioma for the targeted delivery of drugs to glioblastoma. EVs from dogs with glioma showed capacity to traverse the BBB and selectively accumulate within the brain tumor. Overall, this research represents a foundation for the application of autologous EVs to precision glioblastoma treatment, addressing the challenge of BBB penetration and targeting specificity in brain cancer therapy.

Comparison of the Trachea in Normocephalic versus Brachycephalic Cats on the Basis of CT-Derived Measurements.

Tracheal hypoplasia is a major concern in brachycephalic dogs, but there is no consensus for the trachea in brachycephalic cats. We aimed to compare tracheal length and diameter between normo- and brachycephalic cats using computed tomography (CT) image measurements and evaluate their usefulness in tracheostomy planning. A total of 15 normocephalic and 14 brachycephalic cats were included in the study. Tracheas of normocephalic cats were significantly longer compared with brachycephalic cats. No difference was detected in tracheal diameter between normocephalic and brachycephalic cats. Both groups had a lateral diameter significantly larger than the dorsoventral diameter at the level of the cranial end of the manubrium sterni and at the level of the second rib. Normocephalic and brachycephalic cats' tracheas have the same dorsoventral flattening at the level of the cranial end of the manubrium sterni and at the level of the second rib. The location between the 4th and 5th cervical vertebrae seems the best place to perform a tracheostomy in cats due to its round shape and easily accessible anatomical location. No sign of tracheal hypoplasia in brachycephalic cats was detected. Finally, 7 mm appears to be an adequate diameter for the tracheal tubes used to perform feline tracheostomies.

CT measurements of tracheal diameter and length in normocephalic cats.

OBJECTIVES: The aim of this study was to measure the tracheal dimensions of normocephalic cats using CT. METHODS: CT images of 15 client-owned normocephalic cats were retrospectively evaluated to measure the length of the feline trachea. Transverse and vertical inner diameters were measured in five different tracheal regions, and the cross-sectional area of the tracheal lumen was calculated for each point of measurement. Descriptive statistics were applied using a two-tailed t-test. RESULTS: The mean ± SD length of the trachea was 125.13 ± 14.41 mm. Male cats had significantly larger tracheas than female cats. The transverse diameter first increased by 0.94 mm between the most cranial point of measurement and the middle of the trachea. It then decreased by 1.38 mm between the middle of the trachea and the most caudal point of measurement. The vertical diameter decreased by 1.16 mm between the first point of measurement and the penultimate point, and then increased by 0.06 mm between the penultimate point of measurement and the end of the trachea. The two different diameters resulted in an elliptical trachea shape. CONCLUSIONS AND RELEVANCE: The feline trachea was circular only at its cranial and caudal ends, and elliptical with a dorsoventral flattening along the rest of its length. Vertical and transverse diameters varied along the entire length. Tracheal shape differences should be considered when performing permanent tracheostomy, tracheal anastomosis or stenting in cats.

Feline primary nonhematopoietic malignant liver tumours: A multicenter retrospective study (2000-2021).

There is scant literature on primary nonhematopoietic malignant liver tumours (PMLT) in cats. In this retrospective study, medical data of 40 cats diagnosed with PMLT were reviewed over a period of 22 years (2000-2021). The most frequent epithelial tumours were hepatocellular (42.5%) and bile duct carcinomas (32.5%), only six (15%) cats had mesenchymal tumours. The median age was 13 years and clinical signs commonly included ano-/hyporexia (62.5%), apathy/lethargy (52.5%), weight loss (42.5%) and vomiting (35%). At initial diagnosis, metastases were confirmed in 1 (2.5%) and suspected in three (7.5%) cats. Massive was the most frequent morphology (75%). Most intrahepatic tumours were left-sided (54.2%) with the left medial lobe being primarily affected (25%). Extrahepatic tumours were rare (5%). In 34 (85%) cats, liver lobectomy was performed (surgery group), four (10%) were treated palliatively (non-surgery group), and two (5%) received no treatment. Intraoperative complications occurred in 11.8% with four (15.4%) postoperative deaths. Recurrence was detected in 28.6% at a median of 151 days (range, 79-684 days), while postoperative metastases were suspected in 21.4% at a median of 186 days (range, 79-479 days). The median survival time (MST) was significantly longer in cats of the surgery group (375 days) than in the non-surgery group (16 days) (p = .002). MST was 868 days for hepatocellular compared to 270 days for bile duct carcinomas (p = .06). In summary, liver lobectomy is associated with prolonged survival times and good prognosis in cats with hepatocellular, and an acceptable prognosis in cats with bile duct carcinoma.

Increased Sensitivity of Computed Tomography Scan for Neoplastic Tissues Using the Extracellular Vesicle Formulation of the Contrast Agent Iohexol.

Computed tomography (CT) is a diagnostic medical imaging modality commonly used to detect disease and injury. Contrast agents containing iodine, such as iohexol, are frequently used in CT examinations to more clearly differentiate anatomic structures and to detect and characterize abnormalities, including tumors. However, these contrast agents do not have a specific tropism for cancer cells, so the ability to detect tumors is severely limited by the degree of vascularization of the tumor itself. Identifying delivery systems allowing enrichment of contrast agents at the tumor site would increase the sensitivity of detection of tumors and metastases, potentially in organs that are normally inaccessible to contrast agents, such as the CNS. Recent work from our laboratory has identified cancer patient-derived extracellular vesicles (PDEVs) as effective delivery vehicles for targeting diagnostic drugs to patients' tumors. Based on this premise, we explored the possibility of introducing iohexol into PDEVs for targeted delivery to neoplastic tissue. Here, we provide preclinical proof-of-principle for the tumor-targeting ability of iohexol-loaded PDEVs, which resulted in an impressive accumulation of the contrast agent selectively into the neoplastic tissue, significantly improving the ability of the contrast agent to delineate tumor boundaries.

Preliminary Studies on the Intrahepatic Anatomy of the Venous Vasculature in Cats.

Hepatic surgeries are often performed in cats to obtain a disease diagnosis, for the removal of masses, or for the treatment of shunts. Whereas the vascular anatomy of the liver has been studied in dogs, such evidence is lacking in cats. The current study used corrosion casts of portal and hepatic veins and computed tomography (CT) analysis of the casts to identify and describe the intrahepatic anatomy in healthy cat livers (n = 7). The results showed that feline livers had a consistent intrahepatic portal and venous anatomy, with only minor disparities in the numbers of secondary and tertiary branches. The feline portal vein consistently divided into two major branches and not three, as previously described in the literature for cats. The finding of a portal vein originating from the right medial lobe branch leading to the quadrate lobe in 4/7 specimens is a novelty of the feline anatomy that was not previously described in dogs. Partial to complete fusion of the caudate process of the caudate and the right lateral lobe, with a lack of clear venous separation between the lobes, was present in two specimens. These findings allowed a detailed description of the most common intrahepatic venous patterns in cats. Further anatomical studies should be encouraged to confirm the present findings and to investigate the utility of this information in surgical settings.

Feline gastrointestinal eosinophilic sclerosing fibroplasia presenting as a rectal mass.

Case summary: A 9-year-old neutered male cat was referred owing to dyschezia and weight loss. Abdominal CT revealed a heterogeneous mass in the rectum and thickening of one caudal mesenteric lymph node. The mass induced a focal rectal obstruction. Cytological evaluation of fine-needle aspirates showed signs of mixed inflammation for the rectal mass and a reactive lymph node. Because a definite diagnosis was not achieved, complete resection of the mass via a dorsal approach to the rectum was attempted. Histopathology confirmed complete removal and diagnosed feline gastrointestinal eosinophilic sclerosing fibroplasia (FGESF). The cat was treated with psyllium husks and lactulose after surgery. In the postoperative year, the owner reported normal behaviour, food intake and defecation of the patient. Dyschezia reoccurred 14 months after surgery. Imaging revealed recurrence of a rectal mass. Owing to clinical deterioration, the owner elected for euthanasia. Relevance and novel information: This is the first report of rectal FGESF with dyschezia and weight loss as the main clinical signs. The case demonstrates an acceptable outcome for more than 1 year without additional immunosuppressive therapy, and emphasises that FGESF must be considered as a differential diagnosis for rectal masses in cats.

Anaesthetic Management of a Labrador Retriever Undergoing Adrenalectomy for Phaeochromocytoma Excision, a Case Report.

Perioperative management of cases undergoing phaeochromocytoma removal should aim at normalising blood pressure and heart rate, restoring volume depletion, and preventing catecholamine release induced by surgical manipulation. In this case report, a novel pharmacological approach in a dog undergoing surgical tumour excision is described. A 7-year-old 25-kg spayed female Labrador Retriever presented for repeated episodes of generalised weakness, pale mucous membranes, tachycardia, tremor, panting, vomiting, and hypertension over the last month was referred for surgical treatment of a left-sided adrenal tumour with invasion of the caudal vena cava. Severe hypertensive episodes occurred repeatedly, starting early during the anaesthetic period, while clipping and cleaning the abdominal area, and continued intraoperatively when the tumour was handled. Moderate hypotension occurred once the tumour was isolated and worsened during temporary caudal vena cava flow interruption and cavotomy. The patient was treated preoperatively with phenoxybenzamine to prevent hypertensive crises. Intraoperatively, magnesium sulphate and urapidil were used to control blood pressure. This treatment was effective in reducing the magnitude of blood pressure spikes but not sufficient to prevent hypertensive peaks, especially during tumour manipulation. Hypotension was treated with synthetic colloid and crystalloid boli, and noradrenaline continuous infusion. Blood transfusion was performed in response to acute bleeding during cavotomy. The dog recovered successfully from anaesthesia and its quality of life was deemed excellent by the owner at the last follow up, 22 months after surgery. The histopathology confirmed the diagnosis of phaeochromocytoma with an invasion of the phrenicoabdominal vein. In the present case, we obtained a successful outcome but failed to provide haemodynamic stability throughout the procedure.

Total laryngectomy in a cat with a laryngeal peripheral nerve sheath tumor.

OBJECTIVE: To report the surgical technique and outcome of total laryngectomy in a single clinical case. STUDY DESIGN: Case report. ANIMAL: A 5-year-old female spayed domestic shorthair cat. METHODS: A cat presented for acute, severe respiratory distress caused by an invasive laryngeal mass. Incisional biopsy was indicative of sarcoma. Computed tomography of head, neck, and thorax was performed revealing no evidence of metastasis. A total laryngectomy and permanent tracheostomy were performed, and the cat could breathe without difficulties immediately postoperatively. Histopathology confirmed a laryngeal low-grade peripheral nerve sheath tumor (PNST). RESULTS: Surgical margins were free of tumor cells. Surgical revision of the tracheostomy stoma due to obstructive granulation tissue was necessary 24 days after the initial surgery. Nine days after revision surgery, the cat was discharged from the hospital. No evidence of local recurrence or metastasis was detected on repeat computed tomography of the head, neck, and thorax at 6 months, nor on chest radiographs at 12 months postoperatively. At the time of writing (13 months postoperatively), the cat is still alive with a good quality of life. CONCLUSION: Total laryngectomy with permanent tracheostomy allowed the complete removal of an obstructive laryngeal PNST and provided a good quality of life in a cat. CLINICAL SIGNIFICANCE: To the authors' knowledge, this case report represents the first detailed description of the surgical procedure and clinical outcome for a total laryngectomy in a cat.

Characterization of canine epidermal organoid cultures by immunohistochemical analysis and quantitative PCR.

BACKGROUND: Keratinocyte organoids can be used as a tool to evaluate epidermal structure, function and dysfunction. OBJECTIVES: To optimize the canine keratinocyte organoid system and produce organoids that are structurally equivalent to in vivo canine epidermis, in order to enable studies that focus on epidermal diseases and diseases resulting from an impaired epidermal barrier. ANIMALS: Skin biopsies were obtained from five recently euthanized dogs of different breeds with no skin abnormalities. METHODS AND MATERIALS: Cells derived from microdissected interfollicular epidermis were seeded in basement membrane extract and epidermal organoids were grown under different media conditions. Organoids were characterized to assess cell morphology and architecture in haematoxylin and eosin-stained slides and expression of selected epidermal markers (keratin 5, keratin 10, loricrin and filaggrin) by immunohistochemical analysis and quantitative reverse transcription PCR. RESULTS: The selected epidermal markers were expressed in the same epidermal layers in the organoids cultured in expansion medium and differentiation medium as in normal interfollicular epidermis, yet restriction to the distinct layers was best achieved with expansion medium. Comparison of the mRNA expression levels of these markers revealed that relative expression is similar in organoids cultured in expansion medium and normal canine epidermis, while it differs in organoids cultured in differentiation medium. CONCLUSION AND CLINICAL IMPORTANCE: Organoids cultured in expansion medium have an equivalent structure to the interfollicular epidermis and express key marker proteins in similar proportions. Epidermal organoids are therefore a promising in vitro model to study epidermal structure, function and dysfunction.

A complex histopathological challenge: suspicion of an osteoblastoma-like osteosarcoma arising from the second thoracic vertebra in a cat.

BACKGROUND: Reports of osteoblastic tumours are limited to a few case reports in veterinary medicine. Osteoblastoma-like osteosarcoma has been accepted by the World Health Organization as an intermediate form between an osteosarcoma and osteoblastoma. This type of tumour indicates an osteosarcoma, that may resemble osteoblastoma clinically, histologically, and radiologically and have the capability for metastasis. Osteoblastoma-like osteosarcoma has not been described in veterinary medicine so far. CASE PRESENTATION: An eight-year old cat was presented due to progressive ataxia and paraparesis of the pelvic limbs. Imaging confirmed a well-defined, extradural mass originating from the spinous process of the second thoracic vertebra (T2) leading to severe compression of the spinal cord. Decompressive cytoreduction was achieved by removal of the mass after dorsal laminectomy of T1. After recovering from an acute worsening 3.5 weeks after surgery, the cat had an improved neurological status and the dorsal compression was resolved at follow-up 8 months later. A focal contrast enhancing lesion was still evident at the base of T2 spinous process and lung metastasis was additionally suspected. Based on histopathological, radiographic, and clinical features, an "osteoblastoma-like osteosarcoma" was suspected. CONCLUSIONS: To the best of our knowledge, this is the first description of this tumour in veterinary medicine. In addition, this case report highlights the difficulty in the diagnosis and definition of osseous neoplasia in cats and provides a literature review.

An anatomical study of plate-rod fixation in feline tibiae.

OBJECTIVE: To investigate the feasibility of placing bi-cortical cortex (B-cort) or mono-cortical locking screws (M-lock) in a plate-rod construct applied to the feline tibia in combination with different intramedullary (IM) pins. METHODS: Twenty-four feline tibiae of cats were divided into 4 groups, corresponding to IM pin sizes filling approximately 30% (1.0 mm), 40% (1.4 mm), 45% (1.6 mm), and 50% (1.8 mm) of the medullary canal. Computed tomography (CT) was performed to trace potential screw trajectories in each group. A 12-hole, 2.4 mm locking compression plate was then applied on the medial aspect of the tibia. M-lock and B-cort screws were inserted subsequently in each plate hole. Success rates of screw insertion based on CT analysis and cadaveric simulation were compared with screw type, IM pin diameter, and anatomic location as variables. RESULTS: Screw insertion rates were underestimated on CT compared to cadaveric specimens. During cadaveric simulation, B-cort screws could be inserted in all specimens in the 3 most proximal plate holes and in at least 1 of the 3 distal plate holes. The smallest pin size (30%) allowed placement of a greater number of B-cort screws (P < .05) compared to other pins. Fewer B-cort screws could be inserted in the distal diaphyseal region (P < .05) compared to other regions. A total of 99.3% of M-lock screws could be inserted regardless of IMP size. CLINICAL SIGNIFICANCE: Plate rod constructs can include bicortical screws in the proximal and distal metaphysis, and monocortical screws in the diaphysis, combined with an IM pin filling up to 50% of the medullary canal.