Evaluation of a modified radiographic union scale for tibial fractures scoring system in staged bilateral tibial plateau leveling osteotomy procedures and comparison of first and second side radiographic bone healing.

OBJECTIVE: To evaluate the inter- and intra-rater reliability of the tibial plateau leveling osteotomy (TPLO) modified radiographic union scale for tibial fractures (mRUST), a semiquantitative scoring system, as compared with the subjective evaluation of radiographic union for staged TPLOs. STUDY DESIGN: Retrospective study. ANIMALS: Forty-eight dogs, 96 stifles. METHODS: Medical records were reviewed for dogs with bilateral cranial cruciate ligament injuries diagnosed at presentation that underwent staged bilateral TPLOs within 6 months and had both immediate and recheck postoperative radiographs. Radiographs were anonymized, reviewers were blinded, radiographic union was evaluated subjectively, and TPLO mRUST scores were assigned. RESULTS: The subjective evaluation's intra-rater reliability was 0.72 (Kappa 95% CI 0.65-0.79) and inter-rater reliability was 0.33 (Kappa 95% CI 0.28-0.39). The TPLO mRUST scoring system intra-rater reliability was 0.73 (95% CI 0.68-0.78) and inter-rater reliability was 0.56 (95% CI 0.41-0.68). There was no difference in the degree of bone healing quantified by the TPLO mRUST scoring system (95% CI - 0.1-1.2, P = .09) or subjective evaluation (P = .48) between the first and second side TPLOs. The TPLO mRUST scores were positively correlated with subjective healing (r = 0.94, 95% CI 0.92-0.96, P < .0001, and for scores ≥10/12, 99%, 244/246) were subjectively assigned as radiographically healed. CONCLUSION: The TPLO mRUST scoring system improved inter-rater reliability compared to subjective evaluation of radiographic union. CLINICAL SIGNIFICANCE: The TPLO mRUST scoring system should be considered as a semiquantitative supplemental tool for evaluating radiographic union.

Staging canine patients with appendicular osteosarcoma utilizing fluorine-18 fluorodeoxyglucose positron emission tomography/computed tomography compared to whole body computed tomography.

Few studies have investigated the diagnostic performance of fluorine-18-fluorodeoxyglucose (18 F-FDG) positron emission tomography/computed tomography (PET/CT) for staging veterinary patients with appendicular osteosarcoma. The purpose of this study was to evaluate the efficacy of 18 F-FDG-PET/CT compared to whole-body CT (WBCT) for staging canine patients with appendicular osteosarcoma. The 18 F-FDG-PET/CT imaging studies of 66 dogs with appendicular osteosarcoma were anonymized and separated into two detached studies (one with whole body pre- and post-contrast CT images and the other with the whole body pre- and post-contrast CT images with the associated 18 F-FDG-PET overlay). Image assessment was performed retrospectively by five board-certified veterinary radiologists. The radiologists were instructed to assign a predefined categorical score (1-4) to each pre-designated anatomic region based on a devised lesional scoring system. A score of 1 was normal, 2 abnormal but not neoplastic, 3 abnormal and concerning for neoplasia, and 4 abnormal, most likely neoplastic. Overall, the likelihood of detection of '3 or 4' was found to be significantly higher with 18 F-FDG PET/CT when compared to WBCT after adjusting for the effect of evaluator and the subject. Most significantly, 13 osseous lesions concerning for metastasis (scored 3-4) were identified in 10/66 dogs by at least one reviewer on 18 F-FDG PET/CT, which were not identified by any reviewer on WBCT. Additionally, four comorbid neoplastic lesions were identified with 18 F-FDG PET/CT and not with WBCT. The results of this study suggest that 18 F-FDG PET/CT is more efficacious in detecting metastatic and comorbid neoplastic lesions compared to WBCT in dogs with appendicular osteosarcoma.

Characterization of Paeniclostridium sordellii Metalloproteinase-1 in vitro and in an experimental model of infection.

OBJECTIVE: Paeniclostridium sordellii is a pathogen that causes rapidly fatal infections characterized by severe edema, extreme leukemoid reaction and lack of an innate immune response. We recently identified a metalloproteinase of P. sordellii-1 (Mcs1) that cleaves human vascular cell adhesion molecule 1, an adhesion molecule important to hematopoietic precursor retention and leukocyte diapedesis. In the current study, we further characterize Mcs1 activity and investigate its role in pathogenesis. METHODS: Mcs1 peptide cleavage sequence and activity conditions were identified using a semi-quantitative fluorescence-based reporter assay. Additional host targets for Mcs1 protease activity were tested and confirmed by gel electrophoreses and western blots. Finally, Mcs1 knock out (ΔMcs1) and complemented (cMcs1) strains were developed for assessment in our animal model of myonecrosis. RESULTS: Data show that Mcs1 prefers aliphatic amino acid residues, I or L, especially when adjacent to negatively charged or noncharged-polar residues. In vitro, Mcs1 cleaved or partially cleaved human cell adhesion molecules, E-selectin and intracellular adhesion molecule-1 (ICAM-1), and mediators of innate immune infection defense, complement protein-3 and antimicrobial peptide LL-37. In vivo, infection with the ΔMcs1 P. sordellii strain had little effect on animal survival, tissue destruction or circulating white blood cell counts compared to wild type and cMcs1 strains. CONCLUSIONS: Similar to proteolytic virulence factors from other pathogens, Mcs1 is a promiscuous protease that cleaves multiple human-host factors. Despite minimal impact of Mcs1 on the murine model of P. sordellii infection, it is worth considering its role in humans and other animal models.

Flourine-18 fluorodeoxyglucose positron emission tomography imaging exhibits increased SUVmax at the level of the spinal intumescence in normal dogs.

Positron emission tomography (PET) imaging utilizing fluorine-18 labeled fluorodeoxyglucose is a relatively new imaging modality in veterinary medicine that is becoming more common for oncological staging and for musculoskeletal imaging. Thus, it is important to identify the normal variations on PET imaging that may be mistaken for pathology. Variation in standardized uptake values (SUVmax) have been anecdotally identified in the spinal cord of dogs undergoing fluorodeoxyglucose (FDG) PET-CT examinations for oncological staging, with notable increase in SUVmax values identified in the region of the cervical and lumbar spinal intumescences. The aim of this retrospective, analytical study was to compare the SUVmax values at four different locations throughout the spinal cord (C3, C5-T1, T13, and L3-S1) of a group of dogs with no evidence of neurologic disease and compare those findings to histologic specimens from dogs euthanized for unrelated disease. SUVmax values were significantly higher at the cervical and lumbar intumescences in comparison to the control regions (P < .0001 and P < .0001, respectively). Neuronal count and spinal cord gray matter area were also significantly greater at the cervical and lumbar intumescences (neuronal count P = .0025 and P = .0001; area P = .0004 and P = .0009, respectively) while overall neuronal density was lower (P = .003 and P = .028, respectively). We presume the increased SUVmax values at the spinal cord intumescences are the result of overall increased neuron count, increased proportion of gray matter, and increased spinal cord gray matter area. These findings will aid in the interpretation of future PET-CT studies and hopefully prevent the misdiagnosis of spinal cord disease in normal canines.

CT angiographic changes in dogs with acute pancreatitis: A prospective longitudinal study.

Computed tomographic angiography (CTA) has recently been shown to be a useful tool in the diagnosis of acute canine pancreatitis, the identification of pancreatic necrosis, and the detection of sequelae. Evidence of pancreatic necrosis on CTA has been shown to be correlated with a poorer outcome in both humans and dogs and early diagnosis and intervention may improve outcomes. In humans, pancreatic necrosis is typically evident on CTA within 48 h of clinical signs, thus, repeat CTA examinations are often performed to identify pancreatic necrosis that may not have been evident on CTA examinations performed early in the course of disease. Published information investigating the timing of CTA examinations and the use of serial CTA in dogs with acute pancreatitis is lacking. In this prospective, longitudinal study, CTA examinations were performed at the time of hospitalization and repeated 3-5 days later in 11 dogs suffering from acute canine pancreatitis to determine if pancreatic necrosis or sequelae are under diagnosed on examinations performed at the time of hospitalization. Computed tomographic angiography studies were evaluated for changes in pancreatic size, pancreatic contrast enhancement, and peri-pancreatic tissues and vessels. The only statistically significant difference between the initial and repeat CTA examinations was the improvement of fat stranding on the repeat CTA examinations (P < .045). Based on these results, CTA performed at the time of admission is likely adequate in the diagnosis and evaluation of dogs with acute pancreatitis. Repeat CTA examinations are unlikely to add additional information in the absence of worsening clinical signs.

Computed tomographic angiography and ultrasonography in the diagnosis and evaluation of acute pancreatitis in dogs.

BACKGROUND: Acute pancreatitis in dogs is an under-diagnosed disease. Current diagnostic methods are insufficient at identifying sequelae and lack prognostic capability. Computed tomographic angiography (CTA) is accurate for diagnosis and prognostication of pancreatitis in humans. OBJECTIVES: In comparison to ultrasound (US), CTA will (1) better diagnose more severe pancreatitis and sequelae and (2) provide assessment of patient outcome by identification of pancreatic contrast enhancement patterns. ANIMALS: Twenty-six client-owned dogs suspected to have acute pancreatitis. METHODS: US and CTA examinations performed at time of admission were compared to determine the detection of pancreatic changes and sequelae. CTA findings also were compared to outcome indicators for prognosis of dogs with acute pancreatitis. Specific canine pancreatic lipase (cPL) samples were obtained and compared with CTA findings. RESULTS: Ten of 26 dogs had heterogeneous contrast enhancement of the pancreas. Compared to US, CTA better identified portal vein thrombosis (P = .003). Patients with heterogeneous contrast enhancement had longer hospitalization (P = .01), including hospital stays for >5 days (P = .02), had more relapses, and were more likely to have portal vein thrombosis (P = .002). Patients with heterogeneous contrast enhancement had increased spec cPL (P = .006). CONCLUSIONS AND CLINICAL IMPORTANCE: In comparison to US, CTA better identified dogs with more severe acute pancreatitis and those with portal vein thrombosis, factors that may predict longer hospitalization and increased risk of relapse. The presence of heterogeneous contrast enhancement and portal vein thrombosis may change therapy for patients with acute pancreatitis.

Response of phase I and II detoxification enzymes, glutathione, metallothionein and acetylcholine esterase to mercury and dimethoate in signal crayfish (Pacifastacus leniusculus).

Metals and pesticides are common pollutants and the modulation of biomarkers can indicate sub-lethal influences on the physiology of organisms inhabiting impacted aquatic systems. We examined the effects of mercury and the organophosphate pesticide dimethoate on EROD, MROD, glutathione S-transferase (GST), acetylcholine esterase (AChE), metallothionein (MT) and glutathione (GSH) in the signal crayfish (Pacifastacus leniusculus). Crayfish were injected with mercury chloride or dimethoate (0.3, 0.6, 0.9 µg kg-1) and dissected after 72 h. EROD activity in the hepatopancreas did not change in response to mercury chloride treatment but exhibited a dose dependent decrease at all concentrations of dimethoate tested. MROD (hepatopancreas) exhibited a significant decrease at the 0.9 µg kg-1 treatment for both chemicals. GST (hepatopancreas) demonstrated a significant dose dependent decrease at all concentrations of both mercury chloride and dimethoate. AChE (tail muscle) decreased at the 0.6 and 0.9 µg kg-1 concentrations of dimethoate and 0.9 µg kg-1 mercury chloride. In gill tissue, MT increased in response to 0.3 and 0.6 µg kg-1 of mercury chloride but no effect was observed at the 0.9 µg kg-1 concentration of mercury chloride or any concentrations of dimethoate tested. MT did not change in response to mercury or dimethoate in tail tissue. Furthermore, neither chemical modulated GSH concentrations. Our results indicate that, apart from GSH, these markers are sensitive to the pollutants tested and that animals exposed in the wild are potentially compromised in their ability to detoxify environmental contaminants and carry out normal cellular processes.

Experimental identification and computational characterization of a novel extracellular metalloproteinase produced by Clostridium sordellii.

Clostridium sordellii is a lethal pathogen for both animals and humans. Severe capillary leakage, toxic shock syndrome, and an extreme leukemoid reaction (LR), are hallmark features of C. sordellii infections and contribute to its high mortality rate. Here we report the discovery of a previously unknown and uncharacterized metalloproteinase of C. sordellii (referred as Mcs1) that cleaves human vascular cell adhesion molecule (VCAM)-1 in vitro, an adhesion molecule critical to hematopoietic precursor retention and leukocyte diapedesis. We successfully identified the open reading frame encoding Mcs1 within the ATCC 9714 genome and developed an Δmcs1 mutant strain using the ClosTron mutagenesis technology. No VCAM-1 proteolysis was observed from exotoxins collected from mutant strain cultures. Using advanced protein structural modeling and molecular dynamics simulation techniques, the 3D molecular structure and conformational features of Mcs1 were also characterized. Our data demonstrates that Mcs1 proteolytic activity is controlled by the electrostatic interactions between Glu113 and Arg227 residues and the gating motions within its cleft region. This pilot interdisciplinary investigation provided crucial experimental evidence of the existence of Mcs1 in C. sordellii and molecular insights into its 3D structure and proteolytic activity. These findings have the potential to help advance new therapeutics and diagnostics against deadly C. sordellii infections. Follow-up in vitro and in vivo work is under way to further characterize Mcs1 enzymatic kinetics and its role in C. sordellii pathogenesis.

Quantitative Determination of Vinpocetine in Dietary Supplements.

Current United States regulatory policies allow for the addition of pharmacologically active substances in dietary supplements if derived from a botanical source. The inclusion of certain nootropic drugs, such as vinpocetine, in dietary supplements has recently come under scrutiny due to the lack of defined dosage parameters and yet unproven short- and long-term benefits and risks to human health. This study quantified the concentration of vinpocetine in several commercially available dietary supplements and found that a highly variable range of 0.6-5.1 mg/serving was present across the tested products, with most products providing no specification of vinpocetine concentrations.