Recurrence analysis of intraoperative acridine orange-photodynamic therapy for dogs with intranasal tumors

Intraoperative acridine orange-photodynamic therapy (AO-PDT) and cribriform plate irradiation are used to treat canine intranasal tumors. The purpose of this study was to evaluate the effects of AO-PDT on intranasal tumors and the recurrence rate of tumors after this treatment. Treatments with AO-PDT were performed on 38 dogs through a narrow window of the dorsal nasal cavity. The median progression-free interval was 12 mo and recurrence was detected in 21 dogs. Based on computed tomography, recurrence in 16 dogs was biased to the following areas: lateral (n = 10), medial (n = 2), ventral (n = 0), rostral (n = 0), and caudal (n = 8). Side effects were mild and included subcutaneous emphysema and rhinitis. The median survival time was 24 mo. Although AO-PDT with cribriform irradiation is an effective treatment for intranasal tumors, AO-PDT techniques should be improved to treat the nasal cavity more uniformly and thoroughly.     

Surgical therapy of canine nasal tumors: A retrospective study (1982-1986)

The results of surgical therapy in 15 dogs with histologically confirmed nasal tumors were analyzed retrospectively and compared to previous reports. Median survival time for all dogs was seven months. When adjusted for nontumor-related deaths, median survival increased to nine months. These values are two to three times longer than previous reports. To determine possible prognostic indicators, tumor stage, location, and histological type were compared to survival time. Dogs with unilateral nasal tumors had a median survival of 11 months, as compared to three months for dogs with bilateral tumors (p = 0.005). Tumor stage and histological type were not significant factors in comparing survival times.     

High-dose-rate brachytherapy for intranasal tumours in dogs: results of a pilot study

This prospective study describes the feasibility and toxicity of ¹⁹²Iridium high‐dose‐rate (HDR) brachytherapy as an alternative strategy for the treatment of canine intranasal tumours. Fifteen dogs with malignant intranasal tumours were treated twice weekly using a hypofractionated protocol with eight fractions, 5 Gy per fraction, resulting in a total dose of 40 Gy. Acute and chronic adverse side‐effects appeared to be rare. Only 7% of the acute side‐effects and 5% of the chronic were classified as severe (grade 3). Eight dogs showed clinical complete remission, and five dogs had partial remission, with a resolution of tumour‐related symptoms. Magnetic resonance imaging showed a reduced tumour mass in 12 cases. Median survival time was 17 months (range 4–48 months), with four dogs (three without disease) still alive. Median time to recurrence of these dogs was 14 months. In nine dogs, progression or recurrence of the tumour was the cause of death. This study suggests that HDR brachytherapy is feasible and well tolerated.     

A comparison of two dose calculation algorithms—anisotropic analytical algorithm and Acuros XB—for radiation therapy planning of canine intranasal tumors

Although anisotropic analytical algorithm (AAA) and Acuros XB (AXB) are both radiation dose calculation algorithms that take into account the heterogeneity within the radiation field, Acuros XB is inherently more accurate. The purpose of this retrospective method comparison study was to compare them and evaluate the dose discrepancy within the planning target volume (PTV). Radiation therapy (RT) plans of 11 dogs with intranasal tumors treated by radiation therapy at the University of Georgia were evaluated. All dogs were planned for intensity‐modulated radiation therapy using nine coplanar X‐ray beams that were equally spaced, then dose calculated with anisotropic analytical algorithm. The same plan with the same monitor units was then recalculated using Acuros XB for comparisons. Each dog's planning target volume was separated into air, bone, and tissue and evaluated. The mean dose to the planning target volume estimated by Acuros XB was 1.3% lower. It was 1.4% higher for air, 3.7% lower for bone, and 0.9% lower for tissue. The volume of planning target volume covered by the prescribed dose decreased by 21% when Acuros XB was used due to increased dose heterogeneity within the planning target volume. Anisotropic analytical algorithm relatively underestimates the dose heterogeneity and relatively overestimates the dose to the bone and tissue within the planning target volume for the radiation therapy planning of canine intranasal tumors. This can be clinically significant especially if the tumor cells are present within the bone, because it may result in relative underdosing of the tumor.     

A feasibility study of low-dose total body irradiation for relapsed canine lymphoma

Seven client owned dogs with confirmed relapsed lymphoma were enrolled in a prospective feasibility study investigating the effects of low‐dose total body irradiation (LDTBI) delivered in a single 1 Gy fraction. LDTBI for relapsed lymphoma was safe and well tolerated. The only major side‐effect of LDTBI was asymptomatic thrombocytopenia in all dogs. The median platelet nadir was 17 000/μL (range 4000–89 000), which occurred a median of 10 days (range 8–30) post irradiation. Three dogs had short‐term partial responses, two stable disease and two progressive disease (PD). Six dogs were euthanatized for PD, and one dog died while in partial remission. No dogs had clinical complications. Survival analysis was not performed, because the study design did not allow for evaluation of survival time. Larger studies incorporating LDTBI in the induction/consolidation phase of treatment need to be performed to determine the therapeutic efficacy of LDTBI.     

Masitinib for the treatment of canine atopic dermatitis: a pilot study

There is an on-going need to identify medications suitable for the long-term treatment of canine atopic dermatitis (CAD). Masitinib mesilate is a potent and selective tyrosine kinase inhibitor of the c-KIT receptor. A strong relationship exists between the SCF/c-KIT pathway and pathogenesis of CAD, suggesting that masitinib may potentially fulfil the above role. This study reports on an uncontrolled pilot study of masitinib in CAD. Masitinib was administered orally to 11 dogs at a mean dose of 11.0 ± 1.83 mg/kg/day (free base) for 28 days. Treatment response was assessed by evolution of clinical appearance according to a modified version of the Canine Atopic Dermatitis Extent and Severity Index (mCADESI), pruritus scale and surface area of lesions. Masitinib improved CAD with a mean reduction in mCADESI of 50.7 ± 29.8% (95% C.I. = 29.4–72.0; p = 0.0004) at day 28 relative to baseline, with 8/10, 8/10 and 4/10 dogs showing improvement of ≥33%, ≥40% and ≥50%, respectively. Improvement was further evidenced by a decrease in pruritus score and the surface area of lesions. No serious or severe adverse events occurred during this trial, although 6/11 dogs presented with mild to moderate treatment related adverse events. There is sufficient compelling evidence to warrant further investigation.     

Photodynamic therapy for the treatment of intranasal tumors in 3 dogs and 1 cat

Three dogs and 1 cat with intranasal tumors were treated with pyropheophorbide-a-hexyl ether-based photodynamic therapy (PDT). PDT was well tolerated by all the animals, and no adverse effects from photosensitizer injection, such as cutaneous photosensitization, were observed. Facial swelling was observed in all animals after each PDT treatment but resolved spontaneously within 72 hours after treatment. All animals had a decrease in severity of epistaxis, frequency of sneezing, and amount of nasal discharge after PDT. Clinical signs were controlled for variable time, although long-term responses were comparable with radiation therapy in 2 animals. This small case series demonstrates another application for PDT in veterinary medicine. On the basis of these findings. further studies are warranted to define the role of PDT in the management of intranasal tumors in dogs and cats.     

Vinblastine and prednisolone as adjunctive therapy for canine cutaneous mast cell tumors

Twenty-seven dogs with inadequately excised, cutaneous mast cell tumors (MCT; 20 residual microscopic disease, seven marginal excision) were treated with a vinblastine and prednisolone chemotherapeutic protocol. Twenty dogs were available for follow-up examination after 12 months. One dog suffered local recurrence of the tumor, four dogs developed new cutaneous tumors, and one dog had both events. Fourteen dogs were free of MCT. There was no confirmed tumor-related mortality. Although toxicity from the chemotherapy was generally mild, one dog died of sepsis during treatment.     

Ex vivo viability of canine and feline sarcomas: a pilot study

Assay-based chemotherapeutic protocols are common in human gynecologic oncology, most notably for patients with ovarian or breast cancer. The current study examines ex vivo incubation conditions necessary for the assessment of sarcomatous tumor response to potential chemotherapeutic drugs. Slices of sarcomatous tumors were incubated in one of two culture media. Viability indices were measured and compared across time and between media. Neither medium was sufficient to support the growth of sarcomatous tumor tissue slices based on the indices studied. It is likely that sarcomatous tumors require a different approach for ex vivo assessment than their epithelial counterparts. Our long-term goal is to incubate tumor slices with chemotherapeutic agents to predict the in vivo tumor response based on the maintenance or loss of slice viability within this system.     

Cisplatin and doxorubicin combination chemotherapy for the treatment of canine osteosarcoma: a pilot study

Sixteen dogs with histologically confirmed appendicular osteosarcoma were treated by amputation followed by cisplatin and doxorubicin chemotherapy. All dogs began chemotherapy within 24 hours of surgery. Cisplatin was administered at 50 mg/m2 intravenously (IV) concurrent with saline-induced diuresis. Doxorubicin was administered 24 hours later at 15 mg/m2 as a slow IV bolus. This protocol was given on a 21-day cycle for 4 cycles. No dose delays were required, but dose reduction of doxorubicin was required in 2 dogs because of neutropenia. Thoracic radiography was performed every 2 months after completion of therapy to monitor for metastatic disease. Two dogs were still alive and free from disease at the time of last contact (24 and 75 months, respectively). Postmortem examinations were performed on 13 of the 14 dogs that died. Eight of these dogs were euthanized because of metastatic osteosarcoma. Of the remaining 5 dogs, euthanasia was performed because of complications of idiopathic megaesophagus (n = 1), arthritis (n = 2), and hemangiosarcoma (n = 2). The median disease-free interval and survival times were 15.7 and 18 months, respectively. When compared to a historical group of 36 dogs with appendicular osteosarcoma treated with surgery and 4 doses of cisplatin. both disease-free interval and overall survival were significantly longer in the study population (P < .015 and P < .007, respectively).     

Sentinel lymph node mapping of the canine anal sac using lymphoscintigraphy: A pilot study

Sentinel lymph node mapping and biopsy are important parts of oncologic staging in human medicine. Sentinel lymph node mapping enables identification of the first lymph node to receive lymphatic drainage while avoiding unnecessary lymph node dissection. Anal sac adenocarcinoma is the most common malignant neoplasm of the canine perineal area. For dogs with anal sac adenocarcinoma, lympadenectomy and metastasis to the iliosacral lymphocentrum are negative prognostics indicators. The objectives of this prospective, two by two, crossover pilot study were to establish the feasibility of lymphoscintigraphy using Technetium‐99 sulfur colloid of the canine anal sac of healthy dogs, compare two injection techniques, and the time for identification of sentinel lymph nodes using each technique. We hypothesized that both intramural and perimural injections of the canine anal sac would identify similar sentinel lymph node drainage. The sentinel lymph node was identified in all dogs using either technique. Intramural injection of the canine anal sac showed radiopharmaceutical uptake faster than perimural injection technique (P = 0.040). There was concordance between intramual and perimural techniques for the sentinel lymph node identified in 50% of cases. A sacral lymph node was identified as sentinel in three of eight dogs (37.5%). Lymphoscintigraphy of the canine anal sac is safe and feasible in normal dogs; however, the method of injection technique seems to have a significant effect on the sentinel lymph node identified.     

Hypotonic water as adjuvant therapy for incompletely resected canine mast cell tumors: a randomized, double-blind, placebo-controlled study

Objective— To evaluate efficacy of hypotonic water as adjuvant therapy after marginal resection of canine mast cell tumors (MCT).
Study Design— Double-blinded, placebo-controlled, prospective, randomized study.
Animals— Dogs (n=30) with spontaneous, cutaneous, solitary MCT.
Methods— The wound bed of MCT, resected with margins <0.5 cm, was injected with either hypotonic or isotonic water according to a standardized protocol. Follow-up was obtained by clinical examination at 1, 2, 3, 6, and 12 months and annual telephone contact with the owner.
Results— Eighteen dogs were treated with isotonic lactated Ringer's solution and 12 dogs with hypotonic distilled water. All MCT were stage 0 tumors and most grade II. Six tumors (4 isotonic, 2 hypotonic) recurred locally, 3 of these dogs died from disease-related reasons within 4 months. The surviving 3 dogs were alive with a median survival time (ST) of 1092 days. The calculated 2-year recurrence-free rate was 92.7%; the 2-year disease-free rate 79.1%; and the 2-year survival rate 89.5%. No significant differences in local recurrence and ST were observed between treatment groups. Histologic grading was the only significant prognosticator for ST and recurrence-free periods.
Conclusion— No significant differences in local recurrence and ST were observed between adjunctive hypotonic water and placebo treatment after marginal resection of solitary MCT.
Clinical Relevance— Hypotonic water does not decrease the rate of local recurrence in dogs with solitary MCT after marginal surgical excision.     

A pilot study examining a proprietary herbal blend for the treatment of canine osteoarthritis pain

Osteoarthritis is the most common joint disease in dogs. Despite the use of nonsteroidal anti-inflammatory drugs (NSAIDs), many owners seek natural therapies; either to augment the response to NSAIDs, or as a replacement. Substantial research has been directed to investigation of novel therapies. A randomized, double-blinded, controlled study was conducted to assess the efficacy of a herbal remedy for treatment of canine osteoarthritis pain. Client-owned dogs (N = 24) with osteoarthritis were enrolled between 2 veterinary hospitals. Each dog underwent veterinary and owner assessment at 0, 4, and 8 weeks, using the Canine Brief Pain Inventory and Hudson activity scale. Blood was collected for a complete blood (cell) count (CBC) and serum chemistry analysis. The product was deemed to be safe and well-tolerated at the manufacturer recommended dosage, with no significant changes seen in the CBC or serum biochemical analyses. Aside from1 dog that developed gastrointestinal upset, all other dogs tolerated the supplement without complication. The supplement did not statistically improve clinical signs in dogs based on veterinary or owner assessments of lameness. There was a treatment/time effect when assessing veterinary pain scores; however, post-hoc analysis suggests no observable benefit of treatment compared with the placebo group at any time point.     

In vivo acridine orange human spermatozoa staining—A new perspective for RNA detection and spermatozoa morphology evaluation

Significant increases in male infertility and the still unresolved questions on the compatibility and interpretation of current methods in infertility diagnostics call for new protocols. Morphology, genome damage, RNA content and quantity are currently in practice as the major parameters in evaluation of sperm quality. However, results of various methods are not always in mutual concordance. In this study, in vivo acridine orange (AO) staining, which is presently in application in the estimation of genome damage in reticulocytes, was adjusted for spermatozoa staining. Ten men suffering from oligoasthenoteratozoospermia (OAT) and 10 healthy fertile men were analysed using in vivo AO staining. Microscopic analysis was performed by fluorescent and confocal fluorescent microscopy. Our results show that this method preserves spermatozoa membranes, which enables new insight into spermatozoa genome damage, RNA content in residual cytoplasm, damage of neck area with mitochondrion and tail pathology. The introduced method explains the difference between results of sperm DNA fragmentation assay and the globally used AO staining and opens new options for the development of automated systems. In conclusion, the results of our study offer (a) an innovative approach to the analysis of spermatozoa pathology, (b) enable localization and quantification of RNA in residual cytoplasm, (c) a significant contribution to research of aetiology of infertility in men, (d) open new perspectives for the automatization of sperm quality estimation and (e) improve the personalized approach in the selection of in vitro fertilization protocols.     

Evaluation of serum interleukin 2 receptor and beta-2-microglobulin as prognostic factors for canine lymphoma: A pilot study

Interleukin 2 receptor (IL-2R) is released from activated T cell lymphocytes and related to proliferation of B cells and T cells. Beta-2-microglobulin (B2M) is synthesized from all nucleated cells and constitutes a major histocompatibility complex class I antigen. In human medicine, high concentrations of these two factors have been found to be related to prognosis in aggressive non-Hodgkin's lymphoma. In this pilot study, we aimed to assess the correlation between the serum concentration of IL-2R and B2M and the diagnosis and prognosis of canine lymphoma. This study included 8 healthy dogs and 17 dogs with lymphoma. To measure the serum concentration of IL-2R and B2M, a commercial enzyme-linked immunosorbent assay was used. In dogs with lymphoma, IL-2R concentrations were significantly high at the time of diagnosis, but B2M concentrations were not. In relapsed dogs, both IL-2R and B2M concentrations were significantly higher than those in the control and chemotherapy response groups. When the serum concentrations of IL-2R and B2M during chemotherapy were monitored in four relapsed dogs, B2M levels were more closely related with relapse. This study demonstrated that serum IL-2R and B2M concentration can be a diagnostic or prognostic tool for canine lymphoma. Monitoring of serum B2M concentration seems to be useful for predicting relapse.     

Salivary cortisol and behavior in therapy dogs during animal-assisted interventions: A pilot study

Animal-assisted interventions (AAIs) have been associated with positive effects on human psychological and physiological health. Although the perception of quality standards in AAIs is high, only few investigations have focused on potential welfare implications for therapy dogs linked to their performance in AAIs. The standardized program “multiprofessional animal-assisted intervention (MTI)” has been carried out in adult mental health care, significantly improving patients' prosocial behaviors. In the present study, we monitored salivary cortisol and behavioral measures in therapy dogs that participated in MTI group therapy sessions in an in-patient substance abuse treatment facility. Work-related activity (lay, sit, stand, walk, and run), behavior (lip licking, yawning, paw lifting, body shake, tail wagging, and panting), response to human action (taking food treats and obeying commands), and salivary cortisol levels were analyzed over the course of 5 subsequent MTI working sessions in experienced therapy dogs (N = 5), aged 5.4 ± 2.8 years (mean ± standard deviation). Salivary cortisol levels decreased from presession to postsession in sessions 1, 2, and 3. However, only in session 4 and 5, postsession cortisol levels were significantly lower than presession levels (P = 0.043). There was no difference between salivary cortisol levels sampled on a nonworking day at home and work-related levels sampled at the therapy site. None of the behavioral parameters varied significantly over the course of the 5 MTI sessions. Both lip licking (P = 0.038) and body shake (P = 0.021) were positively correlated with the decline in cortisol during session 5. The study results suggest that trained dogs are not being stressed by repeated participation in in-patient substance abuse therapy sessions. Further investigation into the effects of animal-assisted therapy on dogs' physiological markers and behavior is warranted.