Canine preclinical safety evaluation of a multimodal nanoparticle agent (Nanotrast-CF800) for image-guided cancer surgery.

Surgical oncology often requires the use of contrast-enhanced cross-sectional imaging preoperatively to characterize solitary tumours and identify sentinel lymph nodes. Intraoperative optical guidance can effectively aid tissue-sparing tumour excision and locate sentinel lymph nodes. Nanotrast-CF800 (CF800) is a novel dual-modality contrast agent, which co-encapsulates iohexol and indocyanine green (ICG) within a liposomal nanoparticle. It was developed for preoperative and intraoperative imaging of solitary tumours and sentinel lymph node mapping and its efficacy has been demonstrated in preclinical animal models (Zheng et al. 2015). The objective of this study is to evaluate the safety profile of CF800 following intravenous administration in healthy dogs. Six research dogs were randomized into two groups. Group 1 received a low dose (1 mL/kg) and group 2 received a high dose (5 mL/kg). Dogs were placed under general anesthesia and a continuous rate infusion of CF800 was administered based on group allocation. Physiologic parameters including heart rate, respiratory rate, direct arterial blood pressure, cardiac output, and temperature were measured at set time points. Plasma concentrations of iohexol, ICG, and histamine were measured at set time points. Dogs underwent whole body computed tomography scans pre-injection, 2-, and 7-days post-injection (p.i.). Contrast enhancement was measured in select organ systems and great vessels at each time point. There were no significant changes in physiologic parameters following IV infusion of CF800 in all dogs. Plasma iohexol and ICG concentrations peaked at 1 day p.i., while histamine concentrations peaked at 30 minutes p.i. Significant contrast enhancement was noted within the liver, heart, aorta, and caudal vena cava on day 2 p.i., which was significantly different compared to baseline. Prolonged contrast retention within the liver was identified. Intravenous administration of CF800 was safe to use in healthy dogs with no significant systemic adverse effects.

Comparative aspects of targeted sentinel lymph node mapping in veterinary and human medicine: opportunities for future research.

There is a significant overlap in the genetic, metabolic and epigenetic alterations between human and companion animal cancers, including those of the oral cavity, breast, bladder, skin, lungs and pancreas. In many cancer types, the identification and removal of affected lymph nodes are essential for accurate cancer management, including treatment and prognosis. Historically, lymphadenectomy and subsequent radical resection based on regional anatomy, palpation and lymph node aspirates were considered sufficient; however, modern approaches with sentinel lymph node mapping (SLN) mapping have increased the accuracy of surgical decision-making. Preoperative and intraoperative SLN mapping techniques in veterinary patients parallel those used in human medicine. While many of these techniques are highly successful, the main challenges with current methodologies are their sensitivity and specificity for the presence of cancer, which can be overcome via precision medicine and targeted SLN mapping agents. Given the large population of dogs and cats with cancer, the crossover of knowledge between species can help to deepen our understanding of many of these cancers and can be useful in evaluating new drugs and/or therapies. In this review, we discuss SLN mapping techniques in veterinary medicine and the concept of precision medicine as it relates to targeted SLN mapping imaging agents. The large number of companion animals affected by cancer is an underutilized resource to bridge the translational gap and we aim to provide a reference for the use of dogs and cats as a comparative model for human SLN mapping.

Reason for euthanasia in dogs with urothelial carcinoma treated with chemotherapy or radiation therapy or both: A retrospective observational study.

BACKGROUND: Clients want to know the ultimate cause of death in their pet after cancer treatment. The cause of euthanasia and investigation of urinary obstruction in treated dogs with urothelial carcinoma (UC) has not been specifically reported in veterinary literature. HYPOTHESIS/OBJECTIVES: Our hypothesis was that the majority of treated dogs with UC are euthanized secondary to primary tumor factors, such as urinary obstruction. ANIMALS: Fifty-nine client-owned dogs diagnosed with UC. METHODS: Retrospective observational study on clinical signs and disease at euthanasia of dogs with UC treated by radiation therapy or chemotherapy or both. RESULTS: The median overall survival time (OST) of all dogs was 339 days (range, 17-1996; 95% confidence interval [CI], 185-392; interquartile range [IQR], 112-505). Of dogs deemed to have been euthanized because of UC (50/59, 85%), the primary cause was considered to be local progression in 31/50 (62%), most often because of perceived complete or partial urinary obstruction (24/31, 77%). No variables were found to be predictive of urinary obstruction. The overall documented metastatic rate was 56%. In dogs euthanized because of UC, metastasis was deemed to be the cause in 19/50 (38%) dogs. CONCLUSIONS AND CLINICAL IMPORTANCE: Regardless of the type of treatment, UC in dogs has a poor prognosis and there is a continuing need to improve treatments that focus on local control of the primary tumor, given its high contribution to the decision for euthanasia. Proactive management to avoid the high frequency of urinary obstruction may be worthy of future investigation.

Effect of volume and methylene blue on fluorescence intensity and transit of indocyanine green for sentinel lymph node mapping in a simulated feline tumor model.

OBJECTIVE: To compare the effect of volume and solution on transit time and fluorescence intensity (FI) using near-infrared fluorescence imaging (NIRF) in a simulated tumor model in cats. Secondarily, to describe SLN mapping with indocyanine green (ICG) NIRF and report any adverse effects of intradermally injected ICG in cats. ANIMALS: 7 healthy purpose-bred domestic shorthaired male cats. METHODS: Cats were randomly divided into 2 groups (ICG or ICG + methylene blue [MB]). Transit time and FI were determined for 1 or 2 mL solutions injected intradermally in 4 quadrants around a simulated tumor. Following massage, fluorescence intensity was quantified by calculating the corrected total ROI fluorescence using ImageJ software. Cats were monitored for adverse effects up to 4 weeks post-injection. RESULTS: A larger solution volume had decreased transit times to the SLN (P = .001). There was no significant difference in transit times between ICG and ICG-MB. ICG demonstrated a greater FI (P = .001) in the SLN compared to ICG-MB. Methylene blue did not percutaneously fluoresce under NIRF. The volume of the solution did not significantly affect FI. No adverse reactions were reported. CLINICAL RELEVANCE: Increased volume of ICG may aid in rapid percutaneous lymphatic tracking from tumor to SLN. Indocyanine green alone may be preferred over ICG-MB for greater visualization of the SLN. Intradermal injections of ICG and ICG-MB were well-tolerated in healthy cats with no significant complications. Clinical evaluation of this technique in an impaired lymphatic system, as seen in cancer patients, should be assessed in future research.

Evaluating the readability of recruitment materials in veterinary clinical research.

BACKGROUND: Owner comprehension is vital to recruitment and study success, but limited information exists regarding the readability of public-facing veterinary clinical trial descriptions. OBJECTIVES: The current study sought to evaluate the readability of public-facing online veterinary clinical trial descriptions from academic institutions and private referral practices. ANIMALS: None. METHODS: This prospective study assessed readability in a convenience sample of veterinary clinical trial study descriptions using 3 common methods: the Flesch-Kincaid Grade Level (F-K), Flesch Reading Ease Score (FRES), and online Automatic Readability Checker (ARC). Results were compared across specialties and between academic and private institutions. RESULTS: Grade level and readability consensus scores (RCSs) were obtained for 61 online clinical trial descriptions at universities (n = 49) and private practices (n = 12). Average grade-level RCS for study descriptions was 14.13 (range, 9-21). Using Microsoft Word, the FRES score was higher in descriptions from universities compared to private practices (P = .03), and F-K scores were lower in university compared to private practice descriptions (P = .03). FRES (P = .07), F-K (P = .12), and readability consensus (P = .17) scores obtained from ARC were not different between institution types. Forty-eight studies (79%) had RCSs over 12, equivalent to reading material at college or graduate school levels. CONCLUSIONS AND CLINICAL IMPORTANCE: Similar to other areas in veterinary communication, the evaluated veterinary clinical trial descriptions used for advertising and recruitment far exceeded the recommended 6th-grade reading level for medical information. Readability assessments are straightforward to conduct, and ensuring health literacy should be a customary best practice in veterinary medicine and clinical research.

Feasibility of in-home electroencephalographic and actigraphy recordings in dogs.

Introduction: Idiopathic epilepsy is a prevalent neurological disease in dogs. Dogs with epilepsy often present with behavioral comorbidities such as aggression, anxiety, and fear. These behaviors are consistent with pre, post, or interictal behaviors, prodromal changes, seizure-precipitating factors, or absence and focal seizures. The overlap in behavior presentations and lack of objective research methods for quantifying and classifying canine behavior makes determining the cause difficult. Behavioral comorbidities in addition to the task of caring for an epileptic animal have a significant negative impact on dog and caregiver quality of life. Methods: This pilot study aimed to assess the feasibility of a novel technology combination for behavior classification and epileptic seizure detection for a minimum 24-h recording in the dog's home environment. It was expected that combining electroencephalography (EEG), actigraphy, and questionnaires would be feasible in the majority of trials. A convenience sample of 10 community-owned dogs was instrumented with wireless video-EEG and actigraphy for up to 48 h of recording at their caregiver's home. Three questionnaires (maximum 137 questions) were completed over the recording period by caregivers to describe their dog's everyday behavior and habits. Results: Six of the 10 included dogs had combined EEG and actigraphy recordings for a minimum of 24 h. Discussion: This shows that in-home EEG and actigraphy recordings are possible in community-owned dogs and provides a basis for a prospective study examining the same technology combination in a larger sample size.

Outcome of dogs with bone marrow suppression secondary to Sertoli cell tumour.

Sertoli cell tumours are one of the most common canine testicular neoplasia. These tumours are significantly more likely to arise in cryptorchid dogs and are often functional, oestrogen-secreting tumours which can lead to fatal myelotoxicity. The goal of this study was to describe the outcome of dogs with oestrogen-induced bone marrow suppression secondary to Sertoli cell tumours in seven client-owned dogs. Medical records from April 1, 2011 through April 1, 2021 were reviewed to identify dogs that underwent surgical management of a Sertoli cell tumour with documented bone marrow suppression. Overall, 5/7 dogs required transfusion of blood products peri-operatively. Cases 1 and 6 received a transfusion of packed red blood cells (RBC) prior to surgery and case 5 required a transfusion of whole blood. Case 1 also required a transfusion of platelets before surgery. Post-operatively, cases 1 and 2 received packed RBC's and case 6 received two transfusions of whole blood. Case 3 required transfusions of both fresh frozen plasma and platelets post-operatively. All dogs survived to discharge and 6/7 dogs had documented improvement in haematopoietic values. Two dogs remained chronically thrombocytopenic. The median hospital stay was 4 days. One dog died within 4 weeks of surgery from worsening pancytopenia. Survival for greater than 1 year was documented in 4/7 dogs, and one dog was lost to follow-up 4 months post-operatively. One dog remained severely pancytopenic 4 weeks post-operatively and received oral lithium treatment. Improvements in all blood cell lines were observed within the 4 weeks and resolution of pancytopenia within 6 weeks. Historically, the prognosis for dogs with bone marrow suppression secondary to Sertoli cell tumours was guarded to poor. This report documented improved outcomes for dogs that underwent surgery, including one dog that received lithium chloride as treatment for Sertoli cell tumour-induced bone marrow suppression.

Evaluating the Feasibility and Efficacy of a Dual-Modality Nanoparticle Contrast Agent (Nanotrast-CF800) for Image-Guided Sentinel Lymph Node Mapping in the Oral Cavity of Healthy Dogs.

A combination of pre and intraoperative sentinel lymph node (SLN) mapping techniques have been suggested to optimize SLN detection. A novel liposomal nanoparticle, Nanotrast-CF800 (CF800), utilizes computed tomography lymphography (CTL) and near infrared fluorescence imaging (NIRF) for image-guided surgery and SLN mapping. This novel tracer agent has not been evaluated in companion animals. The objective of this study was to evaluate the feasibility and efficacy of CF800 for SLN mapping in the oral cavity of healthy dogs and to report any local adverse effects. Six healthy adult purpose-bred research dogs randomly received either 1 mL (group 1) or 2 mL (group 2) of CF800 injected into the submucosa at the level of the right canine maxillary tooth. CTL and percutaneous NIRF were performed at 1, 3, and 10 min, then 1, 2, 4, 7, and 10 days post-injection (p.i). Overall, both CTL and NIRF identified SLNs in all dogs. The overall peak mean contrast enhancement of the SLNs was 73.98 HU (range 63.45-86.27 HU) at 2 days p.i. Peak fluorescence of the SLN occurred at 1 day p.i. The agent was retained within the SLN for at least 7 days for CTL and 4 days for percutaneous NIRF. No adverse effects were observed. Local administration of CF800 was simple and feasible for the detection of SLNs using CTL+NIRF in the head and neck of healthy dogs and was not associated with significant local adverse events.

DNA mini-barcodes.

Conventional DNA barcoding uses an approximately 650 bp DNA barcode of the mitochondrial gene COI for species identification in animal groups. Similar size fragments from chloroplast genes have been proposed as barcode markers for plants. While PCR amplification and sequencing of a 650 bp fragment is consistent in freshly collected and well-preserved specimens, it is difficult to obtain a full-length barcode in older museum specimens and samples which have been preserved in formalin or similar DNA-unfriendly preservatives. A comparable issue may prevent effective DNA-based authentication and testing in processed biological materials, such as food products, pharmaceuticals, and nutraceuticals. In these cases, shorter DNA sequences-mini-barcodes-have been robustly recovered and shown to be effective in identifying majority of specimens to a species level. Furthermore, short DNA regions can be utilized via high-throughput sequencing platforms providing an inexpensive and comprehensive means of large-scale species identification. These properties of mini-barcodes, coupled with the availability of standardized and universal primers make mini-barcodes a feasible option for DNA barcode analysis in museum samples and applied diagnostic and environmental biodiversity analysis.