Ophthalmic findings and reference values for selected ocular parameters in whooping cranes (Grus americana) and Mississippi sandhill cranes (Grus canadensis pulla).

OBJECTIVE: To establish baseline ophthalmic parameters for an endangered, semi-wild population of healthy whooping cranes (Grus americana) (WHCR) and Mississippi sandhill cranes (Grus canadensis pulla) (SACR). ANIMALS STUDIED: Eighteen WHCR and 16 SACR. PROCEDURES: Ophthalmic examination was performed by a single observer, followed by conjunctival swab collection for aerobic bacterial culture and measurement of tear production (phenol red thread test, PRTT) and corneal diameter (CD) as tolerated. Measurement of the axial globe (AG) length, anterior chamber (AC) depth, lens thickness, vitreous chamber (VC) depth, and pecten length was performed via ocular ultrasound (OUS) as tolerated. RESULTS: Eyelid cicatrization (n = 1 WHCR), keratitis (n = 2 WHCR), incipient cataracts (n = 1 WHCR, n = 4 SACR), and uveal cysts (n = 1 SACR) were identified. Twenty-one bacterial species were cultured from SACR, while 18 bacterial species were cultured from WHCR. SACR under 6 months old had increased PRTT values compared to older SACR (p = .0432). AG length and VC depth of male WHCR were greater than in female WHCR (p = .0045 and p = .0008, respectively). WHCR less than 6 months old had greater AC depth and lens thickness than WHCR over 6 months (p < .001 and p = .0013, respectively). SACR less than 6 months old had greater AC depth and lens thickness than WHCR over 6 months (p < .0001 and p < .0001, respectively). CONCLUSIONS: WHCR and SACR are amenable to complete ophthalmic examination. Age-related differences in PRTT in SACR, sexual dimorphism in WHCR, and age-related differences in AC depth and lens thickness in WHCR and SACR were identified.

Comparison of fibroblast growth factor 19 concentrations between dogs with and without gallbladder mucoceles.

BACKGROUND: Fibroblast growth factor 19 (FGF19) is an enterohepatic hormone the synthesis of which is stimulated by bile acid activation of the nuclear farnesoid X receptor (FXR) in ileal enterocytes. Increased production of FGF19 downregulates hepatocyte bile acid synthesis and gluconeogenesis, while concurrently upregulating hepatocyte glycogenesis and gallbladder (GB) filling. The physiologic impact of this regulatory cycle is illustrated in cholecystectomized humans, in whom the disturbed meal-related flux of GB bile decreases serum FGF19 concentrations. OBJECTIVE: Determine if serum FGF19 concentrations are lower in dogs with clinical GB mucoceles (GBMs) than in control dogs. ANIMALS: Seven dogs with GBM diagnosed using abdominal ultrasonography, biochemical markers, and GB histopathology. Forty-two control dogs without gastrointestinal or hepatobiliary disorders also were evaluated. Health status of controls was assessed by physical examination and diagnostic hematologic and biochemical test results. METHODS: Prospective cross-sectional study to compare fasting plasma or serum FGF19 concentrations between groups. Concentrations of FGF19 were quantified by a commercially available FGF19 ELISA. RESULTS: Concentrations of FGF19 were significantly lower in dogs with clinical GBM (median, 14.0 pg/mL; range, 12.8-67.2) than in control dogs (median, 145.3 pg/mL; range, 36.5-285.1). CONCLUSIONS AND CLINICAL IMPORTANCE: In dogs, GBM is associated with significantly decreased serum FGF19 concentrations. We speculate that this finding reflects compromised GB contraction and decreased enterohepatic circulation of bile flow. Subnormal FGF19 concentrations may influence bile acid synthesis and hepatic metabolism.

Reference Interval Creation for Symmetric Dimethylarginine (SDMA) in Healthy Hispaniolan Amazon Parrots (Amazona ventralis) and Quaker Parrots (Myiopsitta monachus).

Renal disease is often identified as a cause of morbidity and mortality in avian patients. However, currently, early antemortem detection of renal disease in avian patients is difficult. Anatomical and physiological differences between mammals and birds mean the use of commonly employed diagnostic testing (ie, measurement of blood urea nitrogen [BUN] and serum creatinine, urinalysis, and ultrasonography) are either nondiagnostic or difficult to achieve. Symmetric dimethylarginine (SDMA) is considered a more sensitive marker for renal disease in humans, dogs, and cats. However, SDMA has not yet been assessed for diagnostic use in any psittacine species. In this study, we establish reference ranges for SDMA in both Hispaniolan Amazon parrots (Amazona ventralis, HAP) and Quaker parrots (Myiopsitta monachus, QP). Blood was collected from 23 Amazon parrots and 32 Quaker parrots maintained in research facilities. Measurement of SDMA through a commercially available immunoassay (IA-SDMA) as well as creatinine, BUN, uric acid, phosphorus, calcium, sodium, potassium, and chloride were determined through IDEXX Laboratories. Plasma SDMA concentrations ranged from 6 to 15 µg/dL and 3 to 15 µg/dL for the HAP and QP, respectively. Sex was a confounding factor for the QP population, but sex did not have a significant effect on SDMA for the HAP population. No significant correlations were identified between SDMA concentrations and other parameters in either psittacine species. Our results show proof of concept for the IA-SDMA and provide reference intervals for SDMA in HAP and QP. Further investigation is required to determine the validity of this assay and the predictive power of SDMA in the detection of renal impairment for parrots and other common companion birds.

Risk factors for corneal conjunctival grafting failure in dogs at four referral centers in the southern United States (203 patients): 2015-2021.

OBJECTIVE: To report risk factors associated with conjunctival graft failure in dogs at four referral specialty centers. PROCEDURES: Records of 203 dogs (229 eyes) undergoing conjunctival graft repair of ulcerative keratitis at four hospitals from 2015 to 2021 were reviewed. Success was defined as full graft integration with globe retention at the last postoperative evaluation; vision status was reported separately. Factors assessed included patient signalment, ophthalmic examination findings, surgical factors, and follow-up information. RESULTS: Conjunctival graft failure occurred in 11% (25/229) of eyes and was significantly associated with ulcer depth, with corneal perforations having increased odds of graft failure compared with descemetoceles (odds ratio [OR] = 3.22, 95% confidence interval [CI] 1.11-9.32; p = .03) and stromal ulcers (OR = 10.89 [95% CI 1.38-86.18], p = .02). Brachycephalic dogs were significantly more likely than non-brachycephalic dogs to experience graft failure (OR = 5.02 [95% CI 1.42-17.74], p < .01). Surgery on the opposite eye relative to surgeon handedness was significantly associated with an increased risk of graft failure (OR 4.28 [95% CI 1.53-11.94], p < .01). The use of 7-0 and 8-0 suture versus 9-0 (p = .03) and the use of a combined simple continuous and interrupted pattern (p = .03) were significantly associated with an increased risk of graft failure. At the last follow-up, (median 61.5 days), 87% of dogs were visual. CONCLUSIONS: Corneal perforations, surgery on the opposite eye relative to surgeon handedness, suture size and pattern, and brachycephalic conformation were significantly associated with an increased risk of graft failure in this study. These characteristics can be considered when determining prognosis for ulcerative keratitis requiring surgical intervention.

Fluoroscopically measured bronchial collapse in healthy dogs during cough exceeds 25%, and a cutoff of 60% bronchial collapse can be used to distinguish healthy from chronically coughing dogs.

Bronchomalacia, the weakening of bronchial cartilage, is a common cause of cough in dogs. The gold standard for diagnosis of bronchomalacia is bronchoscopy; however, fluoroscopy is often used as a preliminary diagnostic test due to its noninvasiveness. The normal amount of bronchial collapse in healthy dogs using fluoroscopy has not been previously established, with some authors citing <25%. The aims of this prospective and retrospective analytical observational study were to determine the normal amount of airway collapse in healthy animals, the amount of airway collapse in dogs with chronic cough and presumed bronchomalacia, and cut-off values of airway collapse during expiration and cough to distinguish between the two groups. Thoracic fluoroscopy was performed in right and left lateral recumbency during inspiration, expiration, and cough to evaluate the percentage collapse of the trachea and lobar bronchi in 45 healthy and 76 chronically coughing dogs. The bronchi of chronically coughing dogs exhibited a significantly greater amount of collapse during both expiration and cough compared with healthy dogs. Chronically coughing dogs exhibited up to 31.7 ± 26.6% collapse in expiration and 87.5 ± 23.7% collapse during cough, whereas healthy dogs exhibited 14.9 ± 6.1% collapse in expiration and 35.0 ± 13.7% collapse during cough. Based on ROC curves, the authors suggest using cut-off values of 30% during expiration and 60% during cough to distinguish between healthy dogs and those with presumed bronchomalacia. Findings suggest that though healthy dogs can exhibit a greater degree of airway collapse than previously thought, fluoroscopy can be used to distinguish between healthy and presumed bronchomalacic dogs.

The influence of a tension band fixation as an adjunct for arthrodesis of the metacarpophalangeal joint in the horse.

OBJECTIVES: To determine the influence of a stainless-steel cable (SSC) tension band fixation as an adjunct to a locking compression plate (LCP) for arthrodesis of the equine metacarpophalangeal (MCP) joint. STUDY DESIGN: Experimental. An ex vivo biomechanical paired equine cadaver limb study. SAMPLE POPULATION: Five MCP joint pairs were collected from adult Thoroughbred horses, euthanized for reasons unrelated to orthopedic disease. METHODS: Each pair of MCP joints were randomly implanted with either a dorsally placed 5.5 mm LCP and a palmarly placed 2.0 mm SSC or a dorsally placed 5.5 mm LCP alone. Each construct was tested in cyclic loading followed by single cycle to failure in axial compression. Displacement at a target load of 1 kN over 3600 cycles at 1 Hz was recorded prior to single cycle to failure testing. RESULTS: In cyclic testing, displacement was not significantly different between the first and last 5% of testing cycles regardless of construct. Maximum displacement of each construct during cyclic testing was <1.1 mm. In single cycle testing, the observed yield point did not reveal any difference between LCP and LCP-SSC (p = .440). The maximum load at failure was significantly higher in LCP-SSC compared to constructs with the LCP alone (p = .046). CONCLUSION: The addition of the SSC to the LCP did not statistically affect construct displacement during cyclic loading or construct yield load during subsequent single cycle to failure. CLINICAL SIGNIFICANCE: This study provided much needed information regarding the necessity of a tension band SSC application in the arthrodesis of the MCP/MTP joint in horses.

Deletion of PGAM5 Downregulates FABP1 and Attenuates Long-Chain Fatty Acid Uptake in Hepatocellular Carcinoma.

Phosphoglycerate mutase 5 (PGAM5) is a Ser/His/Thr phosphatase responsible for regulating mitochondrial homeostasis. Overexpression of PGAM5 is correlated with a poor prognosis in hepatocellular carcinoma, colon cancer, and melanoma. In hepatocellular carcinoma, silencing of PGAM5 reduces growth, which has been attributed to decreased mitophagy and enhanced apoptosis. Yet in colon cancer, PGAM5's pro-tumor survival effect is correlated to lipid metabolism. We sought to identify whether deletion of PGAM5 modulated lipid droplet accrual in hepatocellular carcinoma. HepG2 and Huh7 PGAM5 knockout cell lines generated using CRISPR/Cas9 technology were used to measure cell growth, cellular ATP, and long-chain fatty acid uptake. Expression of hepatocellular fatty acid transporters, cluster of differentiation 36 (CD36), solute carrier family 27 member 2 (SLC27A2), solute carrier family 27 member 5 (SLC27A5), and fatty acid binding protein 1 (FABP1) was measured by quantitative PCR and Western blot. We found that deletion of PGAM5 attenuates hepatocellular carcinoma cell growth and ATP production. Further, PGAM5 knockout ameliorates palmitate-induced steatosis and reduces expression of FABP1 in HepG2 and Huh7 cell lines. PGAM5's role in hepatocellular carcinoma includes regulation of fatty acid metabolism, which may be related to expression of the fatty acid transporter, FABP1.

Shear wave velocity values measured by 2D-shear wave elastography are not different between awake and anesthetized cats without clinically significant hepatic fibrosis.

Two-dimensional shear wave elastography (2D-SWE) is widely used as a noninvasive method to quantify liver stiffness. In humans, liver stiffness approximates histologic hepatic fibrosis. While histology is the gold standard for diagnosing liver disease, 2D-SWE may be a minimally invasive alternative to biopsy in feline patients. The objectives of this prospective, observational, crossover study were trifold: (1) to assess the feasibility of performing 2D-SWE in awake cats, (2) to determine whether anesthesia altered shear wave velocity (SWV) measurements, and (3) to correlate hepatic stiffness with histologically quantified hepatic fibrosis. Eleven healthy, purpose-bred cats underwent 2D-SWE in awake and anesthetized states. SWV measurements were compared with histologic fibrosis measurements obtained from liver biopsies during the anesthetic period. The mean velocities were not significantly different between awake (1.47 ± 0.18 m/s) and anesthetized (1.47 ± 0.24 m/s) cats. Premedication and anesthetic drugs did not impact mean SWV. There was a higher variability in the SWV values in the awake group. The data points were reliably replicated, with an interquartile range of 0.24 and 0.32 in anesthetized and awake groups, respectively. There was moderate agreement between observers (intraclass correlation coefficient = 0.66). All cats had clinically insignificant fibrosis. There was no correlation between the SWV measurements and the histological fibrosis values. This study demonstrates that 2D-SWE is feasible in awake cats and that the anesthetic protocol employed did not significantly alter mean SWV. This work is the first to histologically validate normal SWV values in cats and show that 2D-SWE cannot differentiate minimal differences in feline hepatic fibrosis.

Relative and Quantitative Characterization of the Bovine Bacterial Ocular Surface Microbiome in the Context of Suspected Ocular Squamous Cell Carcinoma.

The ocular surface microbiome is altered in certain disease states. The aim of this study was to characterize the bovine bacterial ocular surface microbiome (BBOSM) in the context of ocular squamous cell carcinoma (OSCC). The conjunctiva of normal (n = 28) and OSCC (n = 10) eyes of cows aged 2 to 13 years from two farms in Louisiana and Wyoming were sampled using individual sterile swabs. DNA extraction followed by 16S ribosomal ribonucleic acid (rRNA) gene sequencing and real-time polymerase chain reaction (RT-PCR) were performed to, respectively, assess the relative and absolute BBOSM. Discriminant analysis (DA) was performed using RT-PCR data, and relative abundance analysis was performed using 16S rRNA gene sequencing data. The 11 most abundant phyla in both normal and OSCC-affected cows were identified using 16S rRNA gene sequencing analysis. The relative abundance of Euryarchaeota was found to be significantly lower (p = 0.0372) in OSCC eyes compared to normal eyes. Relative abundance differences within and between geographic locations were also identified. Quadratic DA categorized samples as OSCC or normal with 100% sensitivity and 83.3-100% specificity. Relative abundance analysis identified relative BBOSM phylum alterations in OSCC. Quadratic DA can be used to accurately categorize BBOSM from normal and OSCC ocular surface samples.

Lymphopenia predicts reduced survival in canine hepatocellular carcinoma.

Platelet to lymphocyte ratio (PLR) is a prognostic marker in human hepatocellular carcinoma (HCC) however, its utility in canine HCC has not been explored. The aim of the study was to determine if PLR could predict survival outcomes in 42 dogs with HCC. PLR was not a significant predictive factor (p = 0.15) but lymphopenia alone was significantly correlated with a reduced probability of survival (p = 0.024). Further studies are needed to evaluate if peripheral lymphocyte count mirrors that of the tumor microenvironment in canine HCC.

Efficacy of perioperative 2% dorzolamide ophthalmic solution for reduction of postoperative ocular hypertension in dogs following routine phacoemulsification surgery: A prospective clinical trial.

OBJECTIVE: To evaluate the efficacy of 2% dorzolamide ophthalmic solution for reduction of postoperative ocular hypertension (POH) following routine phacoemulsification surgery in dogs. ANIMALS STUDIED: Thirty one dogs (53 eyes) with naturally occurring cataracts undergoing routine phacoemulsification surgery. PROCEDURE(S): A prospective, double-masked, randomized, placebo-controlled study design was utilized. Dogs received 2% dorzolamide ophthalmic solution or saline 1 h prior to surgery then three times daily for 21 days postoperatively in the operated eye(s). Intraocular pressure (IOP) was recorded 1 h prior to surgery and 3 h, 7 h, 22 h, 1 week and 3 weeks postoperatively. Statistical analyses were performed using chi-squared and Mann-Whitney U test with a significance level of p < .05. RESULTS: Postoperative ocular hypertension (IOP ≥25 mmHg, <24 h after surgery) occurred in 28/53 (52.8%) eyes. There was significant reduction in the incidence of POH for eyes receiving dorzolamide (10/26 (38.4%) eyes) versus eyes receiving placebo (18/27 (66.7%) eyes) (p = .0384). Animals were followed for a median of 163 days after surgery. Thirty-seven (37/53 (69.8%)) eyes were visual at final examination and 3/53 (5.7%) globes were enucleated postoperatively. At last follow-up, there was no difference in visual status (p = .9280), need for topical IOP lowering medication (p = .8319) or incidence of glaucoma (p = .5880) based on treatment group. CONCLUSIONS: Perioperative administration of topical 2% dorzolamide reduced the incidence of POH after phacoemulsification in the dogs studied. However, this was not associated with differences in visual outcome, incidence of glaucoma or need for IOP-lowering medications.

Effect of stimulator of interferon genes (STING) signaling on radiation-induced chemokine expression in human osteosarcoma cells.

Cancer cell-intrinsic mechanisms affecting radiation immunomodulation could be exploited to optimize systemic effects of localized radiation. Radiation-induced DNA damage is sensed by cyclic GMP-AMP synthase (cGAS), which ultimately activates stimulator of interferon (IFN) genes (STING). Resultant expression of soluble mediators such as CCL5 and CXCL10 can facilitate recruitment of dendritic cells and immune effector cells into the tumor. The primary objectives of this study were to determine the baseline expression levels of cGAS and STING in OSA cells and evaluate the dependence of OSA cells on STING signaling for eliciting radiation-induced expression of CCL5 and CXCL10. cGAS and STING expression, and CCL5/CXCL10 expression in control cells, STING-agonist treated cells, and cells treated with 5 Gy ionizing radiation were assessed utilizing RTqPCR, Western blot, and ELISA. U2OS and SAOS-2 OSA cells were deficient in STING relative to human osteoblasts (hObs), while SAOS-2-LM6 and MG63 OSA cells expressed equivalent amounts of STING compared to hObs. A dependence on baseline or induced STING expression was observed for STING-agonist, and radiation-induced, expression of CCL5 and CXCL10. This finding was confirmed by performing siRNA knockdown of STING in MG63 cells. These results show that STING signaling is necessary for radiation-induced expression of CCL5 and CXCL10 in OSA cells. Additional studies are necessary to determine whether STING expression in OSA cells in vivo alters immune cell infiltrates after radiation exposure. These data may also have implications for other potentially STING-dependent characteristics such as resistance to oncolytic virus cytotoxicity.

Evaluation of Prothrombin Time and Activated Partial Thromboplastin Time in Native and Citrated Whole Blood in Hispaniolan Amazon Parrots (Amazona ventralis) With a Handheld Point-of-Care Analyzer.

Objective assessment of coagulation in birds is difficult, and traditional methods of measuring prothrombin time (PT) and activated partial thromboplastin time (aPTT) with the use of mammalian reagents have not been validated in birds. Avian-specific reagents must be prepared from brain extract and are not practical for clinical use. The objective of this investigation was to determine whether the InSight qLabs point-of-care analyzer (Micropoint Biotechnologies Inc, Guangdong, China) could measure PT and aPTT in Hispaniolan Amazon parrots (Amazona ventralis) in native and citrated whole blood, and whether the values obtained correlated with clinical appearance and basic hematologic and biochemical parameters from the bird. The qLabs analyzer was able to measure aPTT reliably, but not PT. Activated partial thromboplastin time of citrated blood was significantly different from the aPTT measured from native whole blood (P < 0.001). On the basis of this study, the qLabs machine may be used to measure aPTT, but clinical application between avian species requires further research.

Neoplastic signatures: Comparative proteomics of canine hepatobiliary neuroendocrine tumors to normal niche tissue.

Hepatobiliary neuroendocrine neoplasms are rare cancers in humans and dogs. To date, no large-scale primary hepatobiliary neoplasm omics analyses exist in any species. This limits the development of diagnostic biomarkers and targeted therapeutics. Neuroendocrine cancers are a heterogenous group of neoplasms categorized by their tissue-of-origin. Because the anatomic niche of neuroendocrine neoplasms shapes tumor phenotype, we sought to compare the proteomes of 3 canine hepatobiliary neoplasms to normal hepatobiliary tissue and adrenal glands with the objective of identifying unique protein signatures. Protein was extracted from formalin-fixed paraffin-embedded samples and submitted for tandem mass spectroscopy. Thirty-two upregulated and 126 downregulated differentially expressed proteins were identified. Remarkably, 6 (19%) of the upregulated proteins are correlated to non-hepatobiliary neuroendocrine neoplasia and 16 (50%) are functionally annotated within the exosome cellular compartment key to neuroendocrine signaling. Twenty-six (21%) downregulated proteins are enriched in metabolic pathways consistent with alterations in cancer. These results suggests that characteristic neoplastic protein signatures can be gleaned from small data sets using a comparative proteomics approach.

Buprenorphine has a concentration-dependent cytotoxic effect on equine chondrocytes in vitro.

OBJECTIVE: To investigate the cytotoxic effects of 2 different concentrations of buprenorphine and compare them with bupivacaine and morphine on healthy equine chondrocytes in vitro. SAMPLE: Primary cultured equine articular chondrocytes from 3 healthy adult horses. PROCEDURES: Chondrocytes were exposed for 0 and 2 hours to the following treatments: media (CON; negative control); bupivacaine at 2.2 mg/mL (BUPI; positive control); morphine at 2.85 mg/mL (MOR); buprenorphine at 0.12 mg/mL (HBUPRE); or buprenorphine at 0.05 mg/mL (LBUPRE). Chondrocyte viability was assessed using live/dead staining, water-soluble tetrazolium salt-8 (WST-8) cytotoxic assay, LDH assay, and flow cytometry. All continuous variables were evaluated with a mixed ANOVA with treatment, time, and their interactions as the fixed effects and each horse as the random effect. RESULTS: Buprenorphine showed a concentration-dependent chondrotoxic effect. The viability of chondrocytes was significantly decreased with exposure to HBUPRE and BUPI compared to CON, MOR, and LBUPRE. CLINICAL RELEVANCE: Negligible chondrotoxic effects were observed in healthy cultured equine chondrocytes exposed to 0.05 mg/mL of buprenorphine, whereas higher concentrations (0.12 mg/mL) showed a marked cytotoxic effect. Based on these results, low concentrations of buprenorphine appear to be safe for intra-articular administration. Further evaluation of this dose in vivo is needed before recommending its clinical use.

Comparison of Two Stifle Exploratory Methods Using Mini-Arthrotomy for Diagnosis of Canine Medial Meniscal Pathology: An Ex Vivo Study.

OBJECTIVE: The main aim of this study was to compare the accuracy of stifle exploratory using either a stifle distractor (SD method) or a combination of Hohmann and Senn retractors (HS method) for diagnosing canine medial meniscal tears in cranial cruciate ligament-deficient stifles. STUDY DESIGN: Fifteen pairs of canine cadaveric pelvic limbs were used and cranial cruciate ligament were transected in all stifles. Paired limbs were then randomly assigned to one of five groups based on the tears created in the caudal pole of the medial meniscus: no tear, peripheral detachment, or a variation in three vertical longitudinal tears. A craniomedial mini-arthrotomy was performed by two observers and diagnosis of the medial meniscal status was made utilizing the HS and SD methods. Correct diagnosis of the meniscal tear was compared for both methods and observers. RESULTS: Correct diagnoses were made using the HS and SD methods in 24/30 and 24/30 cases for observer 1 respectively; and in 17/30 and 19/30 cases for observer 2 respectively. There was no significant difference in the correct diagnosis of meniscal tears within each observer between the two methods. CONCLUSION: Both HS and SD methods have equal accuracy for the diagnosis of canine medial meniscal pathology for a board-certified surgeon. Unassisted surgeons using the SD method for the evaluation of the medial meniscus are at no diagnostic disadvantage compared with assisted surgeons utilizing the HS method.

Exploring the association of intratumoral immune cell infiltrates with histopathologic grade in canine mast cell tumors.

Cutaneous canine mast cell tumors (ccMCTs) vary in their biological behavior, treatment, and prognosis, based on their grade. Immune cell infiltration has been associated with prognosis and response to treatments in some human cancers, and immune-targeting therapeutics are increasingly being explored in veterinary oncology. However, currently little is known about the tumor microenvironment (TME) in ccMCTs. Therefore, the objective of this study was to determine the prevalence of T lymphocytes, T regulatory lymphocytes, PD-1+ cells and macrophages in low- and high-grade ccMCTs. Thirty low-grade and 20 high-grade formalin-fixed paraffin-embedded ccMCT samples were included. Immunohistochemistry (IHC) was performed to detect CD3, FOXP3, Iba1, and PD-1 on sequential sections. Three 400x fields with the highest numbers of CD3+ cells were identified for each tumor. The percentage of CD3+, FOXP3+, and Iba1+ cells, and the number of PD-1+ cells, was quantified in each of these three "hot-spot" fields using ImageJ software. Iba1 expression was significantly greater in high-grade compared to low-grade ccMCTs (mean = 12.5% vs. 9.6%, p = 0.043). PD-1 expression was low overall, but a significantly higher number of PD-1-expressing cells was observed in high-grade ccMCTs (median 1 vs. 0, p = 0.001). No significant difference was noted in CD3 and FOXP3 expression between ccMCT grades. Macrophages and PD-1+ cells were more frequent in high-grade, compared to low-grade ccMCTs. Further studies are needed to define the role of macrophages and rare PD-1+ cells in high-grade ccMCTs.

The use of 'trend' statements in veterinary oncology literature.

Statements such as 'trend towards' and 'tended to' in regards to 'statistical significance' are ambiguous and reflect the continued focus on proximity of p-values to .05. By themselves, p-values do not provide a measure of effect size or other information needed to judge clinical importance. The goal of this study was to examine original research articles in the journal Veterinary and Comparative Oncology over 2 years, and describe the use of 'trend' statements. Articles were reviewed for 'trend' and 'tended to' statements, the number of statements made per paper, whether a p-value and/or 95% confidence interval was provided to accompany these statements, and what the p-value was (if provided). We noted 15.8% of articles included at least one 'trend' or 'tended to' statement to describe differences between groups. Specifically, 10.5% of articles used these statements to describe differences without providing the associated p-value, or where p > .05. Of the total 36 'trend' statements noted, six were not accompanied by p-values and eight were associated with p-values >.10. Similarly, four of the 16 'tended to' statements were not accompanied by p-values, and one was associated with a p-value >.10. These data reveal a similar emphasis on p-value proximity in veterinary oncology literature compared to that of human oncology literature. Furthermore, these findings highlight the ambiguity of 'trend' statements and support the development of additional guidelines (e.g., the de-emphasis of p-value proximity to an arbitrary threshold, and effect size reporting) for interpreting results in veterinary oncology research.

Hepatocyte ploidy in cats with and without hepatocellular carcinoma.

BACKGROUND: Domestic cats rarely develop hepatocellular carcinoma. The reason for the low prevalence is unknown. Reductions in hepatocellular ploidy have been associated with hepatic carcinogenesis. Recent work in mice has shown that livers with more polyploid hepatocytes are protected against the development of hepatocellular carcinoma. Hepatocyte ploidy in the domestic cat has not been evaluated. We hypothesized that ploidy would be reduced in peri-tumoral and neoplastic hepatocytes compared to normal feline hepatocytes. Using integrated fluorescence microscopy, we quantified the spectra of ploidy in hepatocellular carcinoma and healthy control tissue from paraffin embedded tissue sections. RESULTS: Feline hepatocytes are predominantly mononuclear and the number of nuclei per hepatocyte did not differ significantly between groups. Normal cats have a greater number of tetraploid hepatocytes than cats with hepatocellular carcinoma. CONCLUSIONS: Total hepatocellular polyploidy in normal cat liver is consistent with values reported in humans, yet cellular ploidy (nuclei per cell) is greater in humans than in cats. Tetraploid cat hepatocytes are predominantly mononuclear.

Influence of repeat abdominal radiographs on the resolution of mechanical obstruction and gastrointestinal foreign material in dogs and cats.

Mechanical obstruction is a common cause of acute vomiting among small animals and radiography is the first line diagnostic tool. Due to many circumstances, surgery may not be an immediate treatment option. The purpose of this retrospective, cross-sectional design study is to determine the incidence of radiographic resolution of mechanical obstruction or gastrointestinal foreign material with medical management. Inclusion criteria included: clinical signs of vomiting, radiographic diagnosis of mechanical obstruction, suspect mechanical obstruction, foreign material suspected or definitively identified on the first set of radiographs and a second set of repeat radiographs within 36 h. Radiographic resolution was dependent on the location of dilation; 18/48 (37.5%) resolved were gastric, 6/35 (17.1%) were small intestinal, and 4/35 (11.4%) had concurrent gastric and small intestinal obstruction. Gastric dilation was more likely to resolve than SI dilation or both gastric and small intestinal dilation (P = .0119). Forty-nine animals without resolution went to surgery, with two developing pneumoperitoneum, and seven requiring intestinal resection, three of which had linear foreign bodies and three had intestinal masses/strictures. In conclusion, only 17.1% of animals with small intestinal dilation and a radiographic diagnosis of mechanical ileus resolved on repeat radiographs, and only 11.4% of animals with both small intestinal and gastric dilation resolved. Gastric dilation alone was more likely to resolve than small intestinal dilation or a combination of both. This may help to guide practitioners in recommendations for medical management of mechanical ileus in the future.