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Protic ruthenium complexes using the dihydroxybipyridine (dhbp) ligand combined with a spectator ligand (N,N = bpy, phen, dop, Bphen) have been studied for their potential activity vs. cancer cells and their photophysical luminescent properties. These complexes vary in the extent of π expansion and the use of proximal (6,6′-dhbp) or distal (4,4′-dhbp) hydroxy groups. Eight complexes are studied herein as the acidic (OH bearing) form, [(N,N)2Ru(n,n′-dhbp)]Cl2, or as the doubly deprotonated (O− bearing) form. Thus, the presence of these two protonation states gives 16 complexes that have been isolated and studied. Complex 7A, [(dop)2Ru(4,4′-dhbp)]Cl2, has been recently synthesized and characterized spectroscopically and by X-ray crystallography. The deprotonated forms of three complexes are also reported herein for the first time. The other complexes studied have been synthesized previously. Three complexes are light-activated and exhibit photocytotoxicity. The log(Do/w) values of the complexes are used herein to correlate photocytotoxicity with improved cellular uptake. For Ru complexes 1–4 bearing the 6,6′-dhbp ligand, photoluminescence studies (all in deaerated acetonitrile) have revealed that steric strain leads to photodissociation which tends to reduce photoluminescent lifetimes and quantum yields in both protonation states. For Ru complexes 5–8 bearing the 4,4′-dhbp ligand, the deprotonated Ru complexes (5B–8B) have low photoluminescent lifetimes and quantum yields due to quenching that is proposed to involve the 3LLCT excited state and charge transfer from the [O2-bpy]2− ligand to the N,N spectator ligand. The protonated OH bearing 4,4′-dhbp Ru complexes (5A–8A) have long luminescence lifetimes which increase with increasing π expansion on the N,N spectator ligand. The Bphen complex, 8A, has the longest lifetime of the series at 3.45 μs and a photoluminescence quantum yield of 18.7%. This Ru complex also exhibits the best photocytotoxicity of the series. A long luminescence lifetime is correlated with greater singlet oxygen quantum yields because the triplet excited state is presumably long-lived enough to interact with 3O2 to yield 1O2. [ABSTRACT FROM AUTHOR]

Splenic hemangiosarcoma has morphological similarities to benign nodular hyperplasia. Computed tomography (CT) texture analysis can analyze the texture of images that the naive human eye cannot detect. Recently, there have been attempts to incorporate CT texture analysis with artificial intelligence in human medicine. This retrospective, analytical design study aimed to assess the feasibility of CT texture analysis in splenic masses and investigate predictive biomarkers of splenic hemangiosarcoma in dogs. Parameters for dogs with hemangiosarcoma and nodular hyperplasia were compared, and an independent parameter that could differentiate between them was selected. Discriminant analysis was performed to assess the ability to discriminate the two splenic masses and compare the relative importance of the parameters. A total of 23 dogs were sampled, including 16 splenic nodular hyperplasia and seven hemangiosarcoma. In each dog, total 38 radiomic parameters were extracted from first‐, second‐, and higher‐order matrices. Thirteen parameters had significant differences between hemangiosarcoma and nodular hyperplasia. Skewness in the first‐order matrix and GLRLM_LGRE and GLZLM_ZLNU in the second, higher‐order matrix were determined as independent parameters. A discriminant equation consisting of skewness, GLZLM_LGZE, and GLZLM_ZLNU was derived, and the cross‐validation verification result showed an accuracy of 95.7%. Skewness was the most influential parameter for the discrimination of the two masses. The study results supported using CT texture analysis to help differentiate hemangiosarcoma from nodular hyperplasia in dogs. This new diagnostic approach can be used for developing future machine learning‐based texture analysis tools. [ABSTRACT FROM AUTHOR]

Glioblastoma (GBM) is the most aggressive type of brain tumor that originates from glioblastoma stem cells (GSCs). In the brain, GSCs are supported by a tumor microenvironment (TME) residing in the perivascular niche and the hypoxic niche. The GBM TME is highly heterogenous and exhibits complex cell‐to‐cell interactions. Three‐dimensional tumorspheres cultured in stem cell‐enriching media is often used as an in vitro model. The GBM tumorspheres retain some of the transcriptional and translational GSC features but often fails to recapitulate intertumor heterogeneity. Here, we developed a simple, matrix‐free, and in vivo‐like GBM organoids (GBOs) using patient‐derived xenograft GBM lines in small‐scale bioreactors. Shear stress was optimized to produce highly reproducible GBOs over 1 mm diameter within 4–5 weeks. GBOs exhibited high stemness and strong cell‐to‐cell interactions compared to conventional tumorsphere cultures. They displayed spatial gradients of hypoxia‐inducible factor 1α positive hypoxic cores where CD133‐positive cells resided and spatially heterogeneous expression of NOTCH and its ligands. We also observed a self‐established, hierarchically organized, and heterogeneous TME by GBM transdifferentiation into endothelial cells, pericytes, and astrocytes. Collectively, we demonstrate the ability to biomanufacture uniformly sized GBOs that recapitulate in vivo GBM TME features that can serve as an improved GBM in vitro model. [ABSTRACT FROM AUTHOR]

To evaluate the effect of kernel and window settings on the assessment of small and complicated vasculature in CT angiographic (CTA) images of kidneys, jejunum with mesentery, and tumors in dogs. ANIMALS 20 healthy dogs and 20 dogs with tumors. PROCEDURES Images from CTA performed previously in dogs were reconstructed with 3 different combinations of kernel and window settings (soft kernel with soft tissue window, soft kernel with bone window, and sharp kernel with bone window), and reconstructed images of the left kidney and the jejunum with the mesentery in healthy dogs and tumors in affected dogs were evaluated by reviewers blinded to the settings. RESULTS For images of kidney and jejunum with mesentery, reviewers' scores for the conspicuity of vascularity in the arterial phase and the differentiation of the organs from the adjacent structures were significantly higher when viewed in bone window (vs soft tissue window) regardless of kernel setting. For images of head and gastrointestinal tumors, reviewers' scores for differentiation of intratumoral vasculature were higher when viewed in sharp kernel with bone window versus other setting combinations. However, the conspicuity of gastrointestinal, hepatic, or splenic tumoral vessels from the adjacent structures had higher reviewer scores for images in soft kernel with soft tissue window, compared with other setting combinations. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that reconstruction of CTA images with sharp kernel combined with bone window settings might have clinical utility in evaluating and planning treatments for dogs with various tumors; however, additional research is warranted to further identify effects of various kernel and window setting combinations on assessments of small and complicated vasculature in larger and more diverse populations of dogs with and without tumors. [ABSTRACT FROM AUTHOR]

Ultrasmall iron oxide nanoparticles (USIONPs) have been recently developed as labeling probes for T2 magnetic resonance imaging contrast agents. However, their use in stem cell tracking has been limited, especially as T1 contrast agents. In this study, we studied the effects of USIONP surface coatings on proliferation, cellular uptake, and multipontency of established and primary neural stem cells (NSCs). USIONPs were functionalized with gluconic acid (GA), tannic acid (TA), and hyaluronic acid (HA) to label the NSCs. All functionalized USIONPs were characterized as T1 contrast agents via relaxivity measurements. Direct functionalization with TA and HA coating promoted NSC proliferation and enhanced cellular uptake in a dose-dependent manner compared to those with GA. Furthermore, HA coating showed enhanced cell proliferation and cellular uptake in primary NSCs depending on the HA molecular weight. Stem cell characteristics were well-maintained, verified by neurosphere formation and gene expression of stemness and differentiation markers. Collectively, we demonstrated that NSC proliferation, cellular uptake, and multipotency can be enhanced using various surface coating strategies of USIONPs. [ABSTRACT FROM AUTHOR]

Objective: This study was performed to evaluate ocular structures using contrast‐enhanced ultrasonography (CEUS) in dogs to assess the feasibility of CEUS for investigating the blood perfusion of canine eyes. Animal studied: Eight purpose‐bred beagles were used. Procedures: Blood perfusion and vascularity of the right eye were evaluated using color Doppler, power Doppler, and CEUS with Sonazoid®. Vascular changes were quantitatively evaluated by measuring peak intensity, time to initial upslope, and time to peak from the ciliary body, iris, choroid, retina, and the retrobulbar region by CEUS. Results: On CEUS images, all parts of the examined ocular structures were markedly enhanced and clearly identified from the adjacent region. After injection, the contrast agent initially flowed to the choroid and retina at 14.2 seconds, then to the ciliary body and iris at 20 seconds. The blood signal reached its peak intensity in the ciliary body at 27.2 seconds (47.4 ± 10.63), in the iris at 31.6 seconds (74.00 ± 41.85), and in the retrobulbar region at 23.4 seconds (149 ± 24.59). The optic nerve was clearly distinguished from the retrobulbar region over 5 minutes after the initiation of CEUS. Significantly, more vascular signals were detected in the ciliary body and iris by CEUS than by color and power Doppler. Conclusion: Blood perfusion of the intraocular structures and the retrobulbar region can be quantitatively and qualitatively analyzed by CEUS. CEUS may be a useful, noninvasive, and sensitive tool for the evaluation of blood perfusion in ocular diseases. [ABSTRACT FROM AUTHOR]

Ruthenium complexes containing a sterically congested metal center can serve as light activated prodrugs through photo-activated chemotherapy (PACT). In this work, we modified PACT agents containing 6,6′-dihydroxybipyridine (6,6′-dhbp) (Papish et al., Inorg. Chem., 2017, 56, 7519) by replacing it with a sterically bulky isoelectronic ligand, 6,6′-dimethoxybipyridine (6,6′-dmbp). The resulting complexes, [(phen)2Ru(6,6′-dmbp)]Cl2 (2OMe, phen = 1,10-phenanthroline) and [(dop)2Ru(6,6′-dmbp)]Cl2 (3OMe, dop = 2,3-dihydro-[1,4]dioxino[2,3-f][1,10]phenanthroline), have been fully characterized and display enhanced quantum yields for blue light triggered photodissociation of 0.024(6) and 0.0030(2), respectively. We have also synthesized 4OH = [(dmphen)2Ru(4,4′-dhbp)]Cl2 wherein dmphen = 2,9-dimethyl-1,10-phenanthroline and 4,4′-dhbp = 4,4′-dihydroxybipyridine. These ligands enhance steric bulk near the metal center and move the hydroxy groups further from the metal center, respectively. Complex 4OH displays a relatively low quantum yield of 0.0014(2). All of the new complexes (2OMe, 3OMe, 4OH) were tested in breast cancer cells (MDA-MB-231) and were non-toxic (IC50 ＞ 100 μM). This has been interpreted in terms of unfavorable log(Do/w) values and furthermore photodissociation alone is insufficient for cytotoxicity. We also report the crystal structures of 4OH and 2OMe, the thermodynamic acidity of complex 4OH, and the redox potentials for all new complexes. [ABSTRACT FROM AUTHOR]

Aim: To investigate whether p38 small-interfering RNA-loaded nanoparticles (p38 siRNA NPs) attenuate spinal nerve ligation (SNL)-induced neuropathic pain in rats by suppressing spinal microglia activation via p38 targeting. Materials & methods: After synthesizing p38 siRNA NPs with sonication, physical characteristics were measured for size and zeta potential. p38 siRNA NPs were then administrated intrathecally into SNL rats if they could reduce pain behavior excellently. Results: p38 siRNA NPs significantly reduced mechanical allodynia as well as microgliosis in the spinal dorsal horns of SNL rats, consistent with a downregulation of p38-related proinflammatory mediators. Conclusion: As p38 in the spinal microglia plays a critical role in neuropathic pain, we expect that p38 siRNA NPs could be a promising tool for the treatment of neuropathic pain. [ABSTRACT FROM AUTHOR]

Objective: This study aimed to find the radiographic characteristics for the detection of hypertrophic cardiomyopathy (HCM) and congestive heart failure (CHF) in cats., Animals: Healthy cats (n = 35) and HCM cats with (21) and without (22) CHF., Methods: On radiography, the cardiac size using vertebral heart score, left atrium enlargement (LAE), and dilation of the pulmonary vessels were assessed. The sensitivity and specificity of the radiographic characteristics regarding LAE were evaluated with the echocardiographic left atrium to aortic root ratio as a reference., Results: In HCM cats, cardiomegaly, LAE, and dilation of the caudal pulmonary artery were found compared with those in healthy cats. The LAE could be predicted using the elevation of the carina with 94.12% specificity but 17.5% sensitivity. When CHF developed, LAE and dilation of the caudal pulmonary vein (PV) were significantly different compared with those in HCM cats without CHF. The distal side of the summated shadow made by the right caudal PV with the ninth rib in HCM cats with CHF was significantly larger than that in HCM cats without CHF and a cut-off value of 5.35 mm was drawn with 75% sensitivity and 100% specificity., Clinical Relevance: Although there was an overlapping of radiographic findings between healthy and HCM cats, radiographic assessment of LAE can be useful for predicting HCM and the distal side of the summated shadow made by the right caudal PV with the ninth rib can predict CHF in HCM cats.

An eight-year-old Maltese dog presented with diarrhea and anorexia. Ultrasonography revealed marked focal wall thickening with loss of layering in the distal ileum. Contrast-enhanced computed tomography (CT) revealed a preserved wall layer with hypoattenuating middle wall thickening. In some segments of the lesion, small nodules protruding toward the mesentery from the outer layer were observed. Histopathology revealed focal lipogranulomatous lymphangitis (FLL) with lymphangiectasia. This is the first report to describe the CT features of FLL in a dog. CT features of preserved wall layers with hypoattenuating middle wall thickening and small nodules can assist in diagnosing FLL in dogs., Competing Interests: The authors declare no conflicts of interest., (© 2023 The Korean Society of Veterinary Science.)

Objective: To investigate typical computed tomography (CT) features for the differentiation of vaginal from the uterine origin in dogs., Animals: 7 healthy Beagles in the prospective study and 5 bitches in the retrospective study., Procedures: In the prospective study, dual-phase CT images were obtained from sexually intact female Beagles (n = 7) during anestrus and estrus. On the CT images, the vagina and uterine horns, body, and cervix were assessed for diameter, attenuation, and contrast enhancement pattern. In the retrospective study, CT features of large vaginal lesions (leiomyoma, leiomyosarcoma, adenocarcinoma, hematocolpos, and Gartner's duct cyst) were assessed in 5 bitches., Results: In normal bitches, the cervix was thicker with strong central enhancement compared to the uterus and vagina. The uterine artery, which enters the mesometrium at the level of the cervix, was clearly visualized and assisted in identifying the cervix. In bitches with large vaginal masses, uterine arteries were displaced by the lesions and could not be used to locate the cervix. In 4/5 dogs with vaginal masses, identification of the cervix allowed the determination of the organ of origin. In 1 dog with adenocarcinoma, CT vaginography was additionally required for determining the origin of mass., Clinical Relevance: Results from this study indicate that the cervix, recognized in CT as focal thickening of the uterus with central enhancement, may be used as a landmark for the differentiation between the uterus and vagina in bitches.

Objective: To investigate the efficacy of a phospholipid-stabilized sulfur hexafluoride microsphere (SHM) contrast agent and water for hydrosonography of the upper portion of the gastrointestinal tract of dogs., Animals: 12 healthy adult Beagles., Procedures: In a crossover study, each dog was anesthetized and underwent noncontrast ultrasonography then hydrosonography following administration of tap water (30 mL/kg) without (water method) or with SHM (0.1 mL; SHM method) via an orogastric tube. There were at least 3 days between hydrosonographic procedures. Wall thickness, wall layer definition, conspicuity of the mucosal-luminal interface, and image quality were evaluated separately in the near and far fields for the gastric cardia, body, and pylorus and descending duodenum and compared among the 3 scanning methods., Results: Mean wall thickness measurements did not differ significantly between the water and SHM methods at any location except the far-field gastric cardia where the mean wall thickness for the SHM method was less than that for the water method. In general, the SHM method improved wall layer definition and conspicuity of the mucosal-luminal interface of structures in the near field, compared with noncontrast method. The water and SHM methods both resulted in superior image quality relative to the noncontrast method for the near-field gastric cardia, far-field gastric cardia, and far-field duodenum., Conclusions and Clinical Relevance: Results indicated that, for dogs, gastrointestinal hydrosonography by use of the SHM method improved wall layer definition and mucosal conspicuity, particularly in near-field images of the upper portion of the gastrointestinal tract.

Objective: To compare qualitative features and quantitative parameters of 2 contrast agents (sulfur hexafluoride microbubbles [SHM; SonoVue] and perfluorobutane [PFB; Sonazoid]) for performance of contrast-enhanced ultrasonography (CEUS) of the pancreas in dogs., Animals: 8 healthy Beagles., Procedures: In a crossover study design, CEUS of the pancreas was performed twice in each dog, once with SHM and once with PFB, in random order with at least 3 days between examinations. The recorded cine images were qualitatively assessed for homogeneity of pancreatic enhancement and conspicuity of the pancreatic signal relative to the background. For the quantitative assessment, circular regions of interest were placed over the pancreatic body, and a time-intensity curve was obtained. For each region of interest, CEUS parameters including peak intensity (PI), time to peak pancreatic enhancement, area under the curve (AUC), and wash-in rate were obtained., Results: The homogeneity of the pancreatic parenchyma was not significantly different between contrast agents. The signal conspicuity relative to background noise was significantly higher with PFB than with SHM. Mean values of PI, wash-in rate, and AUC were significantly higher with PFB than with SHM. Time to peak enhancement was not significantly different between contrast agents., Conclusions and Clinical Relevance: Pancreatic CEUS with SHM and PFB produced similar homogeneity scores, but only PFB provided excellent signal conspicuity. Perfluorobutane produced higher values of PI, wash-in rate, and AUC. Findings indicated that PFB can provide homogeneous and strong enhancement of the pancreas during CEUS in healthy dogs and that pancreatic CEUS parameter values differ with the contrast agent used.

Objective: To evaluate the effect of slice thickness on CT perfusion analysis of the pancreas in healthy dogs., Animals: 12 healthy Beagles., Procedures: After precontrast CT scans, CT perfusion scans of the pancreatic body were performed every second for 30 seconds by sequential CT scanning after injection of contrast medium (iohexol; 300 mg of 1/kg) at a rate of 3 mL/s. Each dog underwent CT perfusion scans twice in a crossover-design study with 2 different slice thicknesses (2.4 and 4.8 mm). Computed tomographic pancreatic perfusion variables, including blood flow, blood volume determined with the maximum slope model, times to the start of enhancement and peak enhancement, permeability, and blood volume determined by Patlak plot analysis, were measured independently by 2 reviewers. The CT perfusion variables were compared between slice thicknesses. Interoperator reproducibility was determined by ICC calculation., Results: Interoperator reproducibility of CT perfusion variable measurements was excellent on 2.4-mm (mean ± SD ICC, 0.81 ± 0.17) and 4.8-mm (0.90 ± 0.07) slice thicknesses, except for time to peak pancreatic enhancement on 2.4-mm-thick slices, which had moderate reproducibility (intraclass correlation coefficient, 0.473). There was no significant difference in measurements of blood flow, blood volume by either method, times to the start and peak of pancreatic enhancement, or permeability between slice thicknesses., Conclusions and Clinical Relevance: Results supported that a thin slice thickness of 2.4 mm can be used for assessment of pancreatic perfusion variables in healthy dogs.

Blood perfusion of skeletal muscle and callus was evaluated using contrast-enhanced ultrasonography (CEUS) in a canine osteotomy model to determine the applicability of CEUS in the assessment of neovascularization during fracture healing and to compare the vascular signals on CEUS between external skeletal fixation and cast-applied dogs. In 6 Beagle dogs, a simple transverse osteotomy was performed at the left tibial shaft and external skeletal fixation (n = 3) or a cast (n = 3) was applied. Radiography, power Doppler ultrasonography (power Doppler), and CEUS were performed until complete union was achieved. On CEUS, vascular changes were quantitatively evaluated by measuring peak intensity (PI) and time to PI in the soft tissue and callus and by counting the vascular signals. Vascular signals from the soft tissue were detected on power Doppler and CEUS on day 2. Significantly more vascular signals were detected by CEUS than by power Doppler. On CEUS, PI in the surrounding soft tissue was markedly increased after the fracture line appeared indistinctively changed on radiography in all dogs. In the cast-applied dogs, vascular signals from the periosteal and endosteal callus were detected on CEUS before mineralized callus was observed on radiography. CEUS was useful in assessing the vascularity of soft tissue and callus, particularly in indirect fracture healing, and provided indications of a normally healing fracture., Competing Interests: The authors declare no conflicts of interest., (© 2020 The Korean Society of Veterinary Science.)

An 18-year-old male Yorkshire Terrier was admitted with a history of neurological signs including dullness and progressive tetraparesis. Physical examination revealed bilaterally symmetrical alopecia and pot-bellied abdomen. Computed tomography and necropsy examination showed a mass across the frontal sinus and cerebral frontal lobe, bilateral adrenocortical hyperplasia, and hepatomegaly. Histopathologically, the tumor lesions consisted of sheets, nests, or cords of small- to medium-sized round-to-polyhedral cells. Adrenal cortex showed bilateral diffuse cellular proliferation, and some hepatocytes showed intracytoplasmic glycogen accumulation. Immunohistochemically, the tumor cells were positive for pancytokeratin, chromogranin-A, neuron-specific enolase, S100, synaptophysin, and thyroid transcription factor-1 but negative for microtubule-associated proein-2 and neurofilament, leading to the diagnosis of neuroendocrine tumor. These tumor cells were also positive for adrenocorticotropic hormone.

In ultrasound/computed tomography (CT) fusion images, ultrasound allows visualization of the target in real time. CT provides a navigation for ultrasound scanning and improves the overview in areas of limited visualization with ultrasound. This study was performed to investigate the feasibility of ultrasound/CT fusion based on an electromagnetic tracking technique using external fiducial markers for canine ocular and periocular regions. In 7 Beagle dogs, contrast-enhanced CT images of the head were obtained with placing external fiducial markers over the frontal region and both sides of the forepaws of the dog. Ultrasonography was performed under a magnetic field by installing a position sensor in the linear probe, without changing the dog's position. The positions of the external fiducial markers were adjusted and matched, based on the CT images. The execution time of co-registration and the distance between the regions of interest and the co-registration points, the frontal bone, cornea, retina, and optic nerve, were estimated. Approximately 60% of external fiducial markers were properly recognized in all dogs. After adjustment, all external fiducial markers were precisely matched. The co-registration execution time was less than 1 min. The distances between the regions of interest and co-registration points were less than 3 mm in all dogs. The electromagnetic tracking technique using external fiducial markers was a simple and applicable method for fusion imaging of a canine head using real-time ultrasonography and CT. This technique can be useful for interventional procedures of retrobulbar and periorbital lesions., Competing Interests: The authors declare no conflicts of interest, (© 2019 The Korean Society of Veterinary Science.)

This study was performed to evaluate the feasibility of ultrasound-guided computed tomography (CT) cholecystography and to establish an optimal protocol. In 8 healthy beagles, CT cholecystography was conducted using four contrast formulas; two dilution ratios (1:1 vs. 1:3) and two total volumes (8 mL vs. 16 mL) of 300 mgI/kg iohexol after ultrasound-guided percutaneous contrast injection into the gallbladder. CT images were obtained at 3, 10, and 30 min after injection and assessed qualitatively and quantitatively. For all contrast formulas, CT cholecystography showed the gallbladder and the intra- and extrahepatic bile ducts. The volume of the gallbladder and size of bile duct were significantly larger when using a volume of 16 mL iohexol than an 8 mL volume regardless of the dilution ratio. The distinction between the common bile duct and duodenum, the filling of the gallbladder, and the patency of bile duct were effectively assessed using a 16 mL volume of contrast agent with either dilution ratio. Beam-hardening artifacts deteriorated CT image quality for visualizing the biliary system when using the dilution ratio of 1:1. Patency of the bile tract could be easily evaluated using a curvilinear planar reconstruction. There was no significant difference in CT scan time among the different conditions. Minor leakage of contrast agent temporarily occurred after contrast injection in 30% of 32 sets of CT cholecystography. Ultrasound-guided percutaneous cholecystography can visualize both gallbladder and biliary tract with minimal artifacts using a contrast agent volume of 16 mL with a 1:3 dilution ratio., Competing Interests: The authors declare no conflicts of interest., (© 2019 The Korean Society of Veterinary Science.)

OBJECTIVE To investigate the effects of respiratory phase, body position, beam center location, and gastric distention on radiographic assessment of liver size in dogs. ANIMALS 12 Beagles. PROCEDURES Liver length and the ratio of liver length to T11 length were determined on lateral radiographic views obtained with various techniques. Images were acquired at maximal expiration or maximal inspiration, with dogs in right or left recumbency, with the beam centered on the caudal border of the scapula or the 13th rib, and after food was withheld or with gastric distention. Effects on organs adjacent to the liver were assessed with CT. Changes of the thoracic cavity during the respiratory cycle were investigated with fluoroscopy. RESULTS Liver length was significantly greater on radiographs obtained at maximal expiration than at maximal inspiration, but there was no increase in the ratio of liver length to T11 length. Body position, beam center location, and gastric distention did not significantly affect liver size. For CT, location of the spleen and stomach and location or size of the liver did not change markedly between right and left recumbency. Fluoroscopy revealed that thoracic width was less at maximal expiration than maximal inspiration. CONCLUSIONS AND CLINICAL RELEVANCE Liver length was greater at maximal expiration than at maximal inspiration because of a smaller thoracic width. Body position, beam center location, and gastric distention did not affect liver length. The ratio of liver length to T11 length was not significantly affected by any of the factors investigated.

OBJECTIVE To evaluate acute changes of the liver by use of shear wave elastography (SWE) and CT perfusion after radiofrequency ablation (RFA). ANIMALS 7 healthy Beagles. PROCEDURES RFA was performed on the liver (day 0). Stiffness of the ablation lesion, transitional zone, and normal parenchyma were evaluated by use of SWE, and blood flow, blood volume, and arterial liver perfusion of those regions were evaluated by use of CT perfusion on days 0 and 4. All RFA lesions were histologically examined on day 4. RESULTS Examination of the SWE color-coded map distinctly revealed stiffness of the liver tissue, which increased from the normal parenchyma to the transitional zone and then to the ablation zone. For CT perfusion, blood flow, blood volume, and arterial liver perfusion decreased from the transitional zone to the normal parenchyma and then to the ablation zone. Tissue stiffness and CT perfusion variables did not differ significantly between days 0 and 4. Histologic examination revealed central diffuse necrosis and peripheral hyperemia with infiltration of lymphoid cells and macrophages. CONCLUSIONS AND CLINICAL RELEVANCE Coagulation necrosis induced a loss of blood perfusion and caused tissue hardening (stiffness) in the ablation zone. Hyperemic and inflammatory changes of the transitional zone resulted in increased blood perfusion. Acute changes in stiffness and perfusion of liver tissue after RFA could be determined by use of SWE and CT perfusion. These results can be used to predict the clinical efficacy of RFA and to support further studies, including those involving hepatic neoplasia.

Focal vertebral bone density changes were assessed in vertebral computed tomography (CT) images obtained from clinically healthy dogs without diseases that affect bone density. The number, location, and density of lesions were determined. A total of 429 vertebral CT images from 20 dogs were reviewed, and 99 focal vertebral changes were identified in 14 dogs. Focal vertebral bone density changes were mainly found in thoracic vertebrae (29.6%) as hyperattenuating (86.9%) lesions. All focal vertebral changes were observed at the vertebral body, except for a single hyperattenuating change in one thoracic transverse process. Among the hyperattenuating changes, multifocal changes (53.5%) were more common than single changes (46.5%). Most of the hypoattenuating changes were single (92.3%). Eight dogs, 40% of the 20 dogs in the study and 61.6% of the 13 dogs showing focal vertebral changes in the thoracic vertebra, had hyperattenuating changes at the 7th or 8th thoracic vertebra. Our results indicate that focal changes in vertebral bone density are commonly identified on vertebral CT images in healthy dogs, and these changes should be taken into consideration on interpretation of CT images.

OBJECTIVE To investigate systemic changes following low-dosage prednisolone administration in dogs. ANIMALS 4 healthy purpose-bred adult male Beagles. PROCEDURES Dogs were administered prednisolone PO at a dosage of 2 mg/kg/d for 2 weeks, 1 mg/kg/d for 4 weeks, and 0.5 mg/kg/d for 3 weeks. Body weight, blood pressure, hepatic size and echogenicity, percentage of vacuolated hepatocytes, serum hepatic enzyme activities and glucose concentration, adrenal gland size, and pancreatic echogenicity were evaluated weekly for 9 weeks. RESULTS The only significant change identified was an increase in hepatic echogenicity, assessed by measuring liver-kidney contrast on ultrasonographic images. Increases in hepatic size and percentage of vacuolated hepatocytes were identified, but values did not differ from baseline values. Similarly, serum hepatic enzyme activities increased, but changes were mild and not significantly different from baseline values. Body weight, pancreatic echogenicity, and serum glucose concentration did not show noticeable changes. Mild systemic hypertension was seen, but blood pressure was not significantly different from the baseline value. Similarly, adrenal gland size steadily decreased during the first 6 weeks and increased again after the prednisolone dosage was decreased to 0.5 mg/kg/d. However, mean adrenal gland size was not significantly different from the baseline value at any time. CONCLUSIONS AND CLINICAL RELEVANCE Results suggested that in dogs, administration of prednisolone at a low dosage was associated with minimal systemic effects.

OBJECTIVE To evaluate dynamic movement of the diaphragm of clinically normal dogs by use of fluoroscopy and to obtain quantitative data of diaphragmatic excursion during spontaneous breathing. ANIMALS 8 healthy male Beagles with no history of respiratory tract disease. PROCEDURES Fluoroscopy was performed during stabilized respiratory conditions. The beam center was located at the level of the diaphragm, and diaphragmatic motion was recorded during 3 respiratory cycles in dogs positioned in left lateral, right lateral, and dorsal recumbency. Extent of excursion of the diaphragmatic cupula and both crura, difference in excursion between the left and right crura, and ratios of the excursions of the diaphragmatic cupula and left and right crura to the length of the eighth thoracic vertebra were determined. RESULTS Diaphragmatic crural excursion was symmetric for dogs in right lateral recumbency, and the crural excursion was approximately three-quarters of the vertebral length; however, crural excursion appeared to be asymmetric for dogs in left lateral recumbency. Mean ± SD difference in excursion between the right and left crura was 22.68 ± 8.68% for left lateral recumbency, 16.63 ± 9.22% for right lateral recumbency, and 18.11 ± 12.96% for dorsal recumbency. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that the fluoroscopic view of a dog positioned in right lateral recumbency may allow better evaluation of the symmetry of diaphragmatic excursion, compared with results for other recumbency positions. This study provided quantitative data on the excursion of diaphragmatic movement observed by use of fluoroscopy in clinically normal Beagles.

Metallo prodrugs that take advantage of the inherent acidity surrounding cancer cells have yet to be developed. We report a new class of pH-activated metallo prodrugs (pHAMPs) that are activated by light- and pH-triggered ligand dissociation. These ruthenium complexes take advantage of a key characteristic of cancer cells and hypoxic solid tumors (acidity) that can be exploited to lessen the side effects of chemotherapy. Five ruthenium complexes of the type [(N,N) 2 Ru(PL)] 2+ were synthesized, fully characterized, and tested for cytotoxicity in cell culture (1 A : N,N = 2,2'-bipyridine (bipy) and PL, the photolabile ligand, = 6,6'-dihydroxybipyridine (6,6'-dhbp); 2 A : N,N = 1,10-phenanthroline (phen) and PL = 6,6'-dhbp; 3 A : N,N = 2,3-dihydro-[1,4]dioxino[2,3-f][1,10]phenanthroline (dop) and PL = 6,6'-dhbp; 4 A : N,N = bipy and PL = 4,4'-dimethyl-6,6'-dihydroxybipyridine (dmdhbp); 5 A : N,N = 1,10-phenanthroline (phen) and PL = 4,4'-dihydroxybipyridine (4,4'-dhbp). The thermodynamic acidity of these complexes was measured in terms of two pK a values for conversion from the acidic form (X A ) to the basic form (X B ) by removal of two protons. Single-crystal X-ray diffraction data is discussed for 2 A , 2 B , 3 A , 4 B , and 5 A . All complexes except 5 A showed measurable photodissociation with blue light (λ = 450 nm). For complexes 1 A -4 A and their deprotonated analogues (1 B -4 B ), the protonated form (at pH 5) consistently gave faster rates of photodissociation and larger quantum yields for the photoproduct, [(N,N) 2 Ru(H 2 O) 2 ] 2+ . This shows that low pH can lead to greater rates of photodissociation. Cytotoxicity studies with 1 A -5 A showed that complex 3 A is the most cytotoxic complex of this series with IC 50 values as low as 4 μM (with blue light) versus two breast cancer cell lines. Complex 3 A is also selectively cytotoxic, with sevenfold higher toxicity toward cancerous versus normal breast cells. Phototoxicity indices with 3 A were as high as 120, which shows that dark toxicity is avoided. The key difference between complex 3 A and the other complexes tested appears to be higher uptake of the complex as measured by inductively coupled plasma mass spectrometry, and a more hydrophobic complex as compared to 1 A , which may enhance uptake. These complexes demonstrate proof of concept for dual activation by both low pH and blue light, thus establishing that a pHAMP approach can be used for selective targeting of cancer cells.

Metastasis is the leading cause of cancer deaths due to the spread of cancer cells through the blood vessels and the subsequent formation of secondary tumors. Metastasizing cancer cells in the human vasculature are called circulating tumor cells (CTCs) and are characterized to express the epithelial cell adhesion molecule (EpCAM). They are further known to survive physiological fluid shear stress (FSS) conditions. However, the effect of FSS on CTC molecular phenotype, such as the epithelial to mesenchymal transition (EMT) and cancer stem cell (CSC) expression, has not been extensively studied. Here, CTCs in FSS are evaluated in an in vitro model system. MCF7 and MDA-MB-231 breast cancer cell lines were grown in adherent and suspension culture media. The cell lines were tested for EMT and CSC genetic and protein markers using qRT-PCR and flow cytometry, respectively. Suspension cells showed a significantly increased EMT signature compared to adherent cells (p<0.05), suggesting that they model cells detaching from primary tumors in vivo. Upon application of FSS, MCF7 and MDA-MB-231 cells did not show a significant change in EMT expression (p>0.05), but there was a statistically significant increase of the CSC population in MCF7 suspension cultures (p<0.05). These results with MCF7 suggest that CTCs can be modeled in vitro as non-adherent cancer cells in FSS and that they show an increased CSC-like signature during circulation, providing new insights to the importance of CSC-targeting strategies when treating metastatic patients.

OBJECTIVE To evaluate effects of position, time in that position, and positive end-expiratory pressure on ground-glass opacity caused by physiologic atelectasis on lung CT images and to determine effects of recumbency position before CT. ANIMALS 6 healthy Beagles. PROCEDURES In a crossover study, dogs were placed in 4 positions (sternal, dorsal, right lateral, and left lateral recumbency) for 2 holding times (30 and 60 minutes). Dogs were then repositioned in sternal recumbency, and CT was performed at 2 positive end-expiratory pressures (0 and 15 mm Hg). Location, distribution, and degree of ground-glass opacities were evaluated on lung CT images. Volume and mean density of the lungs and ground-glass opacities as well as maximum density of ground-glass opacities were evaluated. RESULTS Ground-glass opacities were mainly observed in parts of the lungs that were dependent during the various positions before CT, except for sternal recumbency. Opacities were reversible and decreased or disappeared after lung inflation. Ground-glass opacities were observed most frequently and had greatest severity when dogs were positioned in left lateral recumbency before CT. Ground-glass opacities were negligible for dogs positioned in sternal recumbency before CT. CONCLUSIONS AND CLINICAL RELEVANCE Location and reversibility of ground-glass opacities may help clinicians distinguish whether they are attributable to atelectasis or a result of pathological changes. Dogs should be positioned in sternal recumbency to minimize the occurrence of ground-glass opacities, particularly when several procedures are performed before CT, which increases the time that a dog will remain in the same position.

OBJECTIVE To assess by use of various diagnostic imaging modalities acute changes in livers of healthy dogs after radiofrequency ablation (RFA) and determine the capability of each imaging modality to monitor ablation lesion changes. ANIMALS 6 healthy Beagles. PROCEDURES 12 ablation lesions were created in the liver of the dogs (2 lesions/dog). Ablation lesions were evaluated by use of conventional ultrasonography, strain elastography, and contrast-enhanced ultrasonography immediately after (time 0), 30 to 60 minutes after, and 3 days after RFA, and by use of CT 30 minutes and 3 days after RFA. Three dogs were euthanized shortly after RFA, and the other 3 dogs were euthanized on day 3. Lesion size measured by each imaging modality was compared with necropsy findings. RESULTS Immediately after RFA, clear margins were more visible with elastography and contrast-enhanced ultrasonography than with conventional ultrasonography, which had acoustic shadowing. On triphasic contrast CT, the ablation zone, which indicated necrosis and hemorrhage, was not enhanced and could be measured. Marked enhancement of the periablation rim was observed during the venous phase and was identified as granulation tissue. Size of the ablation area measured on enhanced CT images was strongly correlated with actual lesion size. CONCLUSIONS AND CLINICAL RELEVANCE For dogs of this study, CT was the most reliable method for lesion size determination. Although ultrasonographic imaging measurements underestimated lesion size, all modalities could be used to provide additional real-time guidance for RFA procedures of the liver as well as for other RFA procedures.

OBJECTIVE To evaluate contrast agents for their ability to improve visualization of the colon wall and lumen during CT and ultrasonography. ANIMALS 10 healthy adult Beagles. PROCEDURES Food was withheld from dogs for 36 hours, after which dogs consumed 250 mL of polyethylene glycol solution. Dogs were then anesthetized, a contrast agent (tap water, diluted barium, or air; order randomly assigned) was administered rectally, iodine contrast medium (880 mg of I/kg) was administered IV, and CT and ultrasonography of the colon were performed. After a 1-week washout period, this process was repeated with a different contrast agent until all agents had been evaluated. Two investigators reviewed the CT and ultrasonographic images for colon wall thickness, conspicuity, artifacts, wall layering, and degree of lumen dilation at 4 sites. RESULTS Thickness of the colon wall was greatest in CT and ultrasonographic images with water used as contrast agent, followed by barium and then air. The CT images obtained after water administration had a smooth appearance that outlined the colonic mucosa and had the highest score of the 3 contrast agents for wall conspicuity. Although no substantial artifacts related to any of the contrast agents were identified on CT images, barium- and gas-induced shadowing and reverberation artifacts hindered wall evaluation during ultrasonography. For ultrasonography, the degree of conspicuity was highest with barium in the near-field wall and with water in the far-field wall. In contrast to CT, ultrasonography could be used to distinguish wall layering, and the mucosal and muscular layers were distinct with all contrast agents. CONCLUSIONS AND CLINICAL RELEVANCE Use of water as a contrast agent for both CT and ultrasonography of the colon in dogs compensated for each imaging modality's disadvantages and could be beneficial in the diagnosis of colon disease.

This report describes the usefulness of positron emission tomography-computed tomography (PET-CT) for evaluating recurrent or residual tumors following surgery. CT and 18 F-fluorodeoxyglucose PET-CT were pre- and post-operatively applied to multiple masses in a dog with hemangiosarcoma. The distinction between the left subcutaneous mass and the peritoneum was clarified on pre-operative CT examination, and malignancy was suspected based on PET-CT. A recurrent or residual tumor in the left subcutaneous region was suspected on post-operative PET-CT, and confirmed through histopathologic examination.

Objective: To evaluate whether a low-dosage regimen of prednisolone induces bone loss and whether administration of alendronate sodium prevents glucocorticoid-induced osteopenia in dogs by measuring trabecular bone mineral density (BMD) with quantitative CT., Animals: 8 healthy Beagles., Procedures: In 4 dogs, prednisolone was administered PO at a dosage of 2 mg/kg once daily for 2 weeks, 1 mg/kg once daily for 4 weeks, and 0.5 mg/kg once daily for 3 weeks. In the other 4 dogs, alendronate sodium (2 mg/kg, PO, q 24 h) was whether administered for 9 weeks in addition to the same dosage of prednisolone used in the prednisolone-treated dogs. Before (day 0 [baseline]) and 21, 42, 63, and 150 days after the start of treatment, BMD of the lumbar vertebrae was measured by quantitative CT., Results: BMD in the prednisolone treatment group decreased to 84.7% of the baseline value on day 42, increased to 87.9% on day 63, and recovered to 91.6% on day 150. In the prednisolone-alendronate treatment group, BMD decreased to 91% of the baseline value on day 21, increased to 93.8% on day 63, and then recovered to 96.7% on day 150. Bone mineral density in the prednisolone treatment group was generally lower, albeit not significantly, than that of the prednisolone-alendronate treatment group on each examination day., Conclusions and Clinical Relevance: BMD temporarily decreased after low-dosage prednisolone administration; however, it gradually improved during tapering of the prednisolone dosage. These results have suggested that a low dosage of prednisolone can be used with little concern for development of osteopenia in dogs.

Changes in radial and longitudinal left ventricular (LV) function were investigated in beagles with iatrogenic hypercortisolism. A total of 11 normal dogs were used, and 2 mg/kg prednisone was administered per oral q12 h for 28 days to 7 out of 11 dogs to induce iatrogenic hypercortisolism. Body weight, blood pressure, conventional echocardiography and tissue Doppler imaging (TDI) of normal and iatrogenic hypercortisolism groups were conducted. The myocardial wall velocity of the LV was measured using color TDI and myocardial deformation was determined by the strain and strain rate. Conventional echocardiography revealed that the diastolic LV free wall and interventricular septum in the hypercortisolism group were thickened relative to those in the normal group. The peak early diastolic myocardial velocity and early to late diastolic myocardial velocity ratio of TDI in the hypercortisolism group were significantly lower than those in the normal group. The strain values in the hypercortisolism group were significantly lower than those in the normal group, particularly for longitudinal wall motion. The lower values of myocardium from TDI and strain imaging could be used to investigate subclinical LV systolic and diastolic dysfunction in dogs with the iatrogenic hypercortisolism.

Objective: To determine the effect of region of interest (ROI) setting and slice thickness on trabecular bone mineral density (BMD) measured with quantitative CT in dogs., Animals: 14 healthy Beagles., Procedures: CT of the lumbar vertebrae and a quantitative CT phantom was performed. The BMD of trabecular bone was measured from L1 to L7 in 2 ways in all dogs. First, sequential 9.6-mm-thick CT images were acquired and then CT images were reconstructed into transverse CT images with slice thicknesses of 2.4, 4.8, and 9.6 mm. The obtained images were analyzed by circular ROI and trace ROI methods. Second, lumbar vertebrae were scanned with the installed quantitative CT protocol with a slice thickness of 10 mm and then the CT images were analyzed by installed automatic BMD software., Results: Interclass correlation coefficients of the automatic software (0.975 to 1.0) and the circular method (0.871 to 0.996) were high, compared with those of the trace method (0.582 to 0.996). The BMD measured with the automatic software was not significantly different from that measured with circular ROI and a slice thickness of 9.6 mm. The BMD measured by use of the circular method was not different according to slice thickness., Conclusions and Clinical Relevance: Results obtained by use of automatic software were similar to those obtained by use of more manual methods. The CT images with thinner slice thickness (2.4 and 4.8 mm) could be used in dogs of toy and small breeds to measure lumbar vertebrae BMD to reduce the limitations of the standard 10-mm slice thickness.

A 10-year-old, intact female Yorkshire terrier had multiple pulmonary nodules on thoracic radiography and ultrasonography with no lesions elsewhere. Computed tomography (CT) and positron emission tomography and computed tomography (PET-CT) using 18F-fluorodeoxyglucose (FDG) were performed to identify metastasis and undetected primary tumors. On CT examination, pulmonary nodules had a hypoattenuating center with thin peripheral enhancement, suggesting ischemic or necrotizing lesion. In PET-CT at 47 min after intravenous injection of 11.1 MBq/kg of FDG, the maximum standardized uptake value of each pulmonary nodule was about from 3.8 to 6.4. There were no abnormal lesions except for four pulmonary nodules on the CT and PET-CT. Primary lung tumor was tentatively diagnosed, and palliative therapy using 2 mg/kg tramadol and 2.2 mg/kg carprofen twice per day was applied. After the dog's euthanasia due to deteriorated clinical signs and poor prognosis, undifferentiated pulmonary adenocarcinoma was diagnosed through histopathologic and immunochemistry examination. To the best of the authors' knowledge, this is the first study of CT and PET-CT features of canine pulmonary adenocarcinoma. In this case, multiple pulmonary adenocarcinoma could be determined on the basis of FDG PET-CT through screening the obvious distant metastasis and/or lymph node invasions and excluding unknown primary tumors.

A 5-year-old Shih Tzu was presented with intermittent vomiting and anorexia. Microhepatica and reversed position of the abdominal organs were observed on radiography. Ultrasonographically, portosystemic shunt (PSS) was tentatively diagnosed. Computed tomography (CT) revealed that the distended portal vein drained into the left hepatic vein. The caudal vena cava (CdVC) split postrenally and converged at the renal level. Cranial to this, the azygos continuation of the CdVC was confirmed. In the thorax, a persistent left cranial vena cava (CrVC) was found along with right CrVC. This is the first report of a dog with persistent left CrVC and multiple abdominal malformations. CT angiography was useful in evaluating the characteristics of each vascular anomaly and determining the required surgical correction in this complex case.