Radiographic findings of cardiopulmonary structures can predict hypertrophic cardiomyopathy and congestive heart failure in cats.

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.

Computed tomographic features of focal lipogranulomatous lymphangitis for differentiating from malignant intestinal lesions in a dog.

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.

Ruthenium Complexes with Protic Ligands: Influence of the Position of OH Groups and π Expansion on Luminescence and Photocytotoxicity.

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.

Can CT texture analysis parameters be used as imaging biomarkers for prediction of malignancy in canine splenic tumors?

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.

Computed tomography can differentiate vaginal-origin from uterine-origin lesions in bitches.

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.

Biomanufacturing of glioblastoma organoids exhibiting hierarchical and spatially organized tumor microenvironment via transdifferentiation.

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.

Efficacy of a phospholipid-stabilized sulfur hexafluoride microsphere contrast agent and water for hydrosonography of the upper portion of the gastrointestinal tract in dogs.

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.

Comparison of sulfur hexafluoride microbubbles and perfluorobutane for performance of contrast-enhanced ultrasonography of the pancreas in dogs.

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.

Singlet Oxygen Formation vs Photodissociation for Light-Responsive Protic Ruthenium Anticancer Compounds: The Oxygenated Substituent Determines Which Pathway Dominates.

Ruthenium complexes bearing protic diimine ligands are cytotoxic to certain cancer cells upon irradiation with blue light. Previously reported complexes of the type [(N,N)2Ru(6,6'-dhbp)]Cl2 with 6,6'-dhbp = 6,6'-dihydroxybipyridine and N,N = 2,2'-bipyridine (bipy) (1A), 1,10-phenanthroline (phen) (2A), and 2,3-dihydro-[1,4]dioxino[2,3-f][1,10]phenanthroline (dop) (3A) show EC50 values as low as 4 µM (for 3A) vs breast cancer cells upon blue light irradiation ( Inorg. Chem. 2017, 56, 7519). Herein, subscript A denotes the acidic form of the complex bearing OH groups, and B denotes the basic form bearing O- groups. This photocytotoxicity was originally attributed to photodissociation, but recent results suggest that singlet oxygen formation is a more plausible cause of photocytotoxicity. In particular, bulky methoxy substituents enhance photodissociation but these complexes are nontoxic ( Dalton Trans 2018, 47, 15685). Cellular studies are presented herein that show the formation of reactive oxygen species (ROS) and apoptosis indicators upon treatment of cells with complex 3A and blue light. Singlet oxygen sensor green (SOSG) shows the formation of 1O2 in cell culture for cells treated with 3A and blue light. At physiological pH, complexes 1A-3A are deprotonated to form 1B-3B in situ. Quantum yields for 1O2 (ϕΔ) are 0.87 and 0.48 for 2B and 3B, respectively, and these are an order of magnitude higher than the quantum yields for 2A and 3A. The values for Ï•Δ show an increase with 6,6'-dhbp derived substituents as follows: OMe < OH < O-. TD-DFT studies show that the presence of a low lying triplet metal-centered (3MC) state favors photodissociation and disfavors 1O2 formation for 2A and 3A (OH groups). However, upon deprotonation (O- groups), the 3MLCT state is accessible and can readily lead to 1O2 formation, but the dissociative 3MC state is energetically inaccessible. The changes to the energy of the 3MLCT state upon deprotonation have been confirmed by steady state luminescence experiments on 1A-3A and their basic analogs, 1B-3B. This energy landscape favors 1O2 formation for 2B and 3B and leads to enhanced toxicity for these complexes under physiological conditions. The ability to convert readily from OH to O- groups allowed us to investigate an electronic change that is not accompanied by steric changes in this fundamental study.

Effects of kernel and window setting combinations on assessments of small and complicated vasculature in computed tomography angiographic images of dogs with and without tumors.

OBJECTIVE: 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.

Effect of slice thickness on computed tomographic perfusion analysis of the pancreas in healthy dogs.

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.

Assessment of neovascularization during bone healing using contrast-enhanced ultrasonography in a canine tibial osteotomy model: a preliminary study.

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.

Cellular uptake of protic ruthenium complexes is influenced by pH dependent passive diffusion and energy dependent efflux.

The lipophilic vs. hydrophilic properties of three protic ruthenium compounds were studied as a function of pH. Specifically, we measured Log(Do/w) values for [(N,N)2Ru(6,6'-dhbp)]2+ complexes (where N,N = 2,2'-bipyridine (1A), 1,10-phenanthroline (2A), 2,3-dihydro-[1,4]dioxino[2,3-f][1,10]phenanthroline (3A) and 6,6'-dhbp is the diprotic 6,6'-dihydroxy-2,2'-bipyridine ligand) from pH 4.0 to 8.0. This study allowed us to demonstrate that as the ligand is deprotonated at higher pH values the resulting neutral charge on the complex improves its lipophilic properties. Thus, improved uptake by passive diffusion is expected with protic ligands on Ru(II). Furthermore, cellular studies have demonstrated that passive diffusion is the dominant pathway for cellular uptake. However, metabolic inhibition has also shown that energy dependent efflux reduces the amount of the ruthenium complex (as measured by mean fluorescence intensity) in the cells. These compounds have been shown by fluorescence microscopy to accumulate in the nuclei of cancer cells (MCF7, MDA-MB-231, and HeLa). Taken together, this data shows that uptake is required for toxicity but uptake alone is not sufficient. The greatest light activated toxicity appears to occur in breast cancer cell lines with relatively moderate uptake (MCF7 and MDA-MB-231) rather than the cell line with the greatest uptake of complex 3A (normal breast cell line MCF-10A).

Electromagnetic tracking-based ultrasound/computed tomography fusion imaging in dogs: preliminary application to ocular and periocular regions.

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.

Canine adrenocorticotropic hormone-producing sinusoidal neuroendocrine tumor associated with Cushing's disease.

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.

Ultrasound-guided transhepatic computed tomography cholecystography in beagle dogs.

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.

Targeting Hyaluronan Interactions for Glioblastoma Stem Cell Therapy.

Even with rigorous treatments, glioblastoma multiforme (GBM) has an abysmal median survival rate, greatly due to the drug-resistant glioblastoma stem cell (GSC) population. GSCs are known to remodel their microenvironment, but the precise role of extracellular matrix components hyaluronic acid (HA) and hyaluronidases (HAases) on the GSC population is still largely unknown. Our objective was to determine how HAase can sensitize GSCs to chemotherapy drugs by disrupting the HA-CD44 signaling. GBM cell line U87-MG and patient-derived D456 cells were grown in GSC-enriching media and treated with HA or HAase. Expressions of GSC markers, HA-related genes, and drug resistance genes were measured via flow cytometry, confocal microscopy, and qRT-PCR. Proliferation after combined HAase and temozolomide (TMZ) treatment was measured via WST-8. HA supplementation promoted the expression of GSC markers and CD44 in GBM cells cultured in serum-free media. Conversely, HAase addition inhibited GSC gene expression while promoting CD44 expression. Finally, HAase sensitized GBM cells to TMZ. We propose a combined treatment of HAase and chemotherapy drugs by disrupting the stemness-promoting HA to target GSCs. This combination therapy shows promise even when temozolomide treatment alone causes resistance.

Contrast-enhanced ultrasonography for evaluation of blood perfusion in normal canine eyes.

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.

Sterically demanding methoxy and methyl groups in ruthenium complexes lead to enhanced quantum yields for blue light triggered photodissociation.

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.

Effect of radiographic technique on assessment of liver size in Beagles.

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.