Anti-tumor activity of a recombinant measles virus against canine lung cancer cells.

Canine primary lung cancer with metastasis has a poor prognosis with no effective treatment. We previously generated a recombinant measles virus (MV) that lost binding affinity to a principal receptor, SLAM, to eliminate its virulence as a new cancer treatment strategy. The virus, rMV-SLAMblind, targets nectin-4, recently listed as a tumor marker, and exerts antitumor activity against nectin-4-positive canine mammary cancer and urinary bladder transitional cell carcinoma cells. However, the effectivity of rMV-SLAMblind for other types of canine cancers is still unknown. Here we evaluated the antitumor effect of rMV-SLAMblind to canine lung cancer. Nectin-4 is expressed on three canine lung cancer cell lines (CLAC, AZACL1, AZACL2) and rMV-SLAMblind was able to infect these cell lines. CLAC cells showed reduced cell viability after virus infection. In the CLAC xenograft nude mouse model, intratumoral administration of rMV-SLAMblind significantly suppressed tumor growth. In rMV-SLAMblind-treated mice, natural killer cells were activated, and Cxcl10 and Il12a levels were significantly increased in comparison with levels in the control group. In addition, the depletion of NK cells reduced the anti-tumor effect. To understand difference in efficacy among canine lung cancer cell lines, we compared virus growth and gene expression pattern after virus treatment in the three canine lung cancer cell lines; virus growth was highest in CLAC cells compared with the other cell lines and the induction of interferon (IFN)-beta and IFN-stimulated genes was at lower levels in CLAC cells. These results suggested that rMV-SLAMblind exhibits oncolytic effect against some canine lung cancer cells and the cellular response after the virus infection may influence its efficacy.

Glucoregulatory factors in canine hepatocellular carcinoma and leiomyosarcoma with non-islet cell tumour hypoglycaemia.

INTRODUCTION: Hypoglycaemia caused by malignant tumours other than insulinoma is referred to as non-islet cell tumour hypoglycaemia (NICTH), which may be caused by hepatocellular carcinoma (HCC) and leiomyosarcoma (LMS) in veterinary medicine. However, the pathogenetic mechanism of NICTH remains unclear. Therefore, this study aimed to evaluate the gene-expression levels of glucoregulatory factors in canine HCC and LMS accompanied by hypoglycaemia. MATERIALS & METHODS: Four patients (three with HCC and one with LMS) exhibiting hypoglycemia were included in the hypoglycemic (H) group, whereas ten patients not exhibiting hypoglycemia were in the non-hypoglycaemia (NH) group. The preoperative and postoperative blood glucose and serum insulin-like growth factor-2 (IGF-2) levels, as well as the expression of genes involved regulating blood glucose levels were analysed. RESULTS: Compared with the NH group, the H group exhibited significantly decreased blood-glucose levels, which increased to normal values after surgery. Compared with the NH group, the H group exhibited significantly increased gene expression of insulin-like growth factor 1, IGF-2, and insulin-like growth factor binding protein 3 in the tumours. Conversely, expression of genes encoding glucoregulatory factors including insulin, gastric inhibitory polypeptide and glucagon was not observed. Serum IGF-2 levels were significantly higher in the H group compared with that in the control group (healthy dogs) and NH group. In two cases in the H group, serum IGF-2 levels decreased after tumour resection. CONCLUSION: These results suggest that NICTH development in dogs with HCC and LMS is mechanistically associated with IGF-2 overexpression and elevated serum IGF-2 levels.

Effect of different contrast medium injection protocols for analysis of hepatic computed tomography perfusion in healthy dogs.

OBJECTIVE: To evaluate the hepatic CT perfusion (CTP) for determining the appropriate protocol for the dual-input maximum-slope model in dogs. ANIMALS: 5 healthy dogs. PROCEDURES: Each dog underwent CTP with different contrast medium administration protocols. Combinations of three different injected doses of iohexol (450, 600, and 750 mg/kg) and injection durations (5, 10, and 15 seconds) were used. The CT values at the aorta, portal vein, and hepatic parenchyma were measured to create a time-density curve, and CTP parameters were measured simultaneously on each hepatic lobe using a 320-row multidetector CT scanner. RESULTS: The maximum peak enhancement at the aorta, portal vein, and hepatic parenchyma was greater with the 750-mg/kg dose than with the 450-mg/kg dose. With an injection duration of 15 seconds, the aortic enhancement peak was less, and the arrival time at the aortic enhancement peak was longer compared to that with a 5-second injection duration. The CTP parameters in the caudate process of the caudate lobe and left lateral lobe differed with different injection durations. The CTP parameters in the caudate process of the caudate lobe, left lateral lobe, and right lateral lobe differed with varying injected doses. CLINICAL RELEVANCE: Our study demonstrated that rapid administration of the contrast medium was required for quantitative analysis of hepatic CTP in healthy dogs. The CTP parameters differed with respect to the contrast medium administration protocol, and it was necessary to administer the contrast medium within a fixed duration and at a fixed dose to evaluate CTP correctly.

Serum vascular endothelial growth factor in dogs with various proliferative diseases.

Angiogenesis plays an important role in the proliferation and metastasis mechanisms of malignant tumors. Vascular endothelial growth factor (VEGF), a group of cytokines that contribute to angiogenesis and vasculogenesis. This study aimed to investigate the serum VEGF-A concentrations in dogs with various proliferative diseases. A total of 202 dogs that were histopathologically diagnosed with proliferative diseases were included in the study. Serum VEGF-A concentrations were measured using enzyme-linked immunosorbent assay. Median serum VEGF-A concentrations in dogs were as follows: healthy dogs, 4 pg/ml [0-21 pg/ml]; hepatocellular carcinoma, 30 pg/ml [0-158 pg/ml, P=<0.001]; hepatocellular adenoma, 32 pg/ml [0-49 pg/ml, P=0.003]; hepatic nodular hyperplasia, 18 pg/ml [0-51 pg/ml, P=0.595]; adrenal pheochromocytoma, 32 pg/ml [0-187 pg/ml, P=<0.001]; adrenocortical carcinoma, 32 pg/ml [3-161 pg/ml, P=0.002]; adrenocortical adenoma, 27 pg/ml [0-106 pg/ml, P=0.005]; colorectal adenocarcinoma, 36 pg/ml [0-75 pg/ml, P=0.002]; colorectal adenoma, 43 pg/ml [0-48 pg/ml, P=0.144]; inflammatory colorectal polyps, 37 pg/ml [0-111 pg/ml, P=<0.001]; pulmonary adenocarcinoma, 35 pg/ml [4-107 pg/ml, P=0.002]; pulmonary histiocytic sarcoma, 35 pg/ml [0-131 pg/ml, P=0.016]; and follicular thyroid carcinoma, 35 pg/ml [0-106 pg/ml, P=0.009]. The serum VEGF-A concentrations were significantly higher in dogs with neoplastic lesions compared to healthy dogs, except for colorectal adenoma. High serum VEGF-A concentrations were observed in dogs with proliferative diseases. The present study suggests that angiogenesis-inhibiting therapy, which targets VEGF-A, may be useful for canine neoplastic diseases.

Comparison of outcomes between medical and surgical treatment in dogs with prostatic adenocarcinoma: a retrospective study.

BACKGROUND: Prostatic cancer is uncommon in dogs. Dogs with prostatic carcinoma have been reported to have a poor prognosis. Information regarding prognosis with various surgery options as well as prognosis with surgical vs. medical treatment is lacking. This retrospective study compares the outcomes of medical management to surgical treatment in dogs with prostatic adenocarcinoma and assesses the surgical outcomes of patients who underwent total prostatectomy (TP) and prostatocystectomy (TPC). The medical records of 41 dogs with prostatic adenocarcinoma, between February 2008 and June 2019, were reviewed for information on signalment, clinical signs in the initial evaluation, preoperative diagnostic imaging findings, treatment type (non-surgical or surgical), surgery type, postoperative complications, adjunctive medical therapy, and survival time. The dogs were divided into non-surgical (n = 12) or surgical (n = 29) groups. The surgical group was subdivided into the TP (n = 20) and TPC (n = 9) subgroups. RESULTS: Age was not significantly different between the surgical (median 13.1 years [8.4-15.4] years) and the non-surgical groups (median 10.8 [7.7-15.3] years). Body weight (BW) was also not significantly different between the surgical (median 6.8 kg [2.4-34.5 kg]) and non-surgical groups (median 6.4 kg [3.7-9.12 kg]). The overall median survival time (MST) from the initial evaluation was significantly longer in the surgical than in the non-surgical group (337 vs. 90.5 days). The postoperative MST was significantly longer in the TP group than in the TPC subgroup (510 vs. 83 days). As TPC was performed in cases of tumor progression, its postoperative complications were severe, resulting in a shorter MST. Ten (50%) and 6 patients (30%) in the TP subgroup postoperatively showed mild and severe urinary incontinence, respectively, whereas all patients in TPC subgroup did show severe incontinence. CONCLUSION: Results of the study suggest that surgical treatment of prostatic carcinoma results in longer survival times over medical management alone. In particular, TP might be recommended for improving survival time and quality of life in canine prostatic adenocarcinoma that does not infiltrate the bladder. Early detection is key for a survival advantage with surgical treatment.

Neutrophilic leucocytosis induced by granulocyte colony-stimulating factor and interleukin-6 in canine primary lung adenocarcinoma.

BACKGROUND: Neutrophilic leucocytosis as a paraneoplastic syndrome may occur in dogs with lymphoma, renal carcinoma, rectal polyps and metastatic fibrosarcoma. However, the information on canine lung adenocarcinoma with neutrophilic leucocytosis is lacking. OBJECTIVE: This study aimed to describe the clinical features and cytokine profiles of canine patients with primary lung adenocarcinoma and neutrophilic leucocytosis. METHODS: Two dogs (cases #1 and #2), each with a solitary lung adenocarcinoma, were included. Both cases had leucocytosis and underwent lung lobectomy. The resected tumours were analysed for the expression of granulocyte colony-stimulating factor (G-CSF), granulocyte macrophage colony-stimulating factor (GM-CSF) and interleukin-6 (IL6) by quantitative real-time PCR compared with normal lung tissues. RESULTS: At the initial examination, neither patient had any clinical signs or fever. White blood cell count (WBC) was 58,300/µl and 32,900/µl in cases #1 and #2, respectively. The gene expression of G-CSF increased 6.7-and 19.7-fold in cases #1 and #2, respectively. The gene expression of IL6 markedly increased (30-fold) in case #1, whereas it increased slightly (1.9-fold) in case #2. On the other hand, that of GM-CSF was slightly changed in both cases. The WBC count postoperatively decreased to within the normal range in both cases. The postoperative survival times were 347 and 118 days in cases #1 and #2, respectively. CONCLUSIONS: This study describes G-CSF and IL6 producing lung adenocarcinoma associated with neutrophilic leucocytosis in dogs. Canine patients with pulmonary adenocarcinomas that have elevated G-CSF and IL6 levels may have a guarded prognosis. Further investigations are needed to clarify the prognosis of canine cytokine-producing lung adenocarcinoma.

Antitumor activity of an oncolytic measles virus against canine urinary bladder transitional cell carcinoma cells.

The prognosis of canine transitional cell carcinoma (TCC) of urinary bladder is generally poor because it is difficult to diagnose at early stages and conventional therapies, such as surgical resection and/or chemotherapy, are often not curative treatments. Based on our previous report that recombinant measles virus (rMV-SLAMblind) therapy could be a new treatment for canine mammary tumor, the applicability of rMV-SLAMblind in canine urinary bladder TCC was examined in this study. A canine TCC cell line was established from a TCC patient dog by transplanting a piece of the tumor mass into an immunodeficient mouse and then isolating the primary TCC cells from the grown tumor mass. The primary cultured cells, named TCC-NU1, express nectin-4, a receptor for rMV-SLAMblind infection. The rMV-SLAMblind infected TCC-NU1 cells, and dose-dependently showed cell cytotoxicity. Moreover, intratumoral administration of rMV-SLAMblind in a xenograft model bearing TCC-NU1 cells significantly suppressed the tumor growth reducing the endpoint mass of tumors in treated mice compared to control mice. These results suggest that virotherapy with rMV-SLAMblind be a new candidate therapy for canine TCC.

Utility of contrast-enhanced ultrasound in differential diagnosis of adrenal tumors in dogs.

This prospective case study aimed to clarify the clinical significance of contrast-enhanced ultrasound (CEUS) for the differential diagnosis of canine adrenal tumors. Forty-three client-owned dogs with adrenal tumors were included. All dogs underwent CEUS, which was evaluated qualitatively and quantitatively. The peak signal intensity (PI), time to peak signal intensity (TPI), mean transit time (MTT), upslope, and downslope were calculated for each time-intensity curve. The histopathological diagnosis of each resected mass was compared with the CEUS findings and parameters. Enhancement distribution, vascularity, tortuous nourishing vessels, enhancement pattern, and late-phase enhancement did not differ significantly between adrenal cortical adenoma (CA), adenocarcinoma (CAC), and pheochromocytoma (PHEO) in qualitative assessment. In PHEO, the TPI was significantly more rapid compared with that in CA (P=0.0287) and CAC (P=0.0404). The MTT in PHEO was significantly shorter than that in CA (P=0.0016) and CAC (P=0.0003). Upslope in PHEO was larger than that in CAC (P=0.0406). Downslope in PHEO was significantly larger than that in CA (P=0.0048) and CAC (P=0.0018). A receiver operating characteristic curve analysis demonstrated that the area under the MTT curve yielded 0.91 for distinguishing PHEO from adrenocortical tumors in dogs; an MTT cut-off value less than 6,225 msec yielded a sensitivity of 69%, specificity of 94%, and likelihood ratio of 12.46. CEUS appears to be clinically applicable for the differential diagnosis between cortical and medullary origins of primary adrenal tumors in dogs.

Computed tomographic-derived measurements of shunt fraction and hepatic perfusion in dogs with a single extrahepatic portosystemic shunt in a clinical setting.

OBJECTIVE: To evaluate the clinical feasibility and usefulness of measuring shunt fraction (SF) and hepatic perfusion with CT in dogs with a single extrahepatic portosystemic shunt (EPSS). ANIMALS: 36 client-owned dogs with EPSS. PROCEDURES: Dogs with EPSS referred for treatment between February 2016 and May 2017 were eligible for the clinical trial. Shunt type, SF, and hepatic perfusion were determined in each dog with a 320-row multidetector CT scanner, and surgical treatment was performed by a single veterinary surgeon. Differences in results between dogs grouped according to age (< 3 years vs ≥ 3 years), shunt type, and subgroups (eg, clinical signs and surgical procedure) were analyzed, and correlations between the SF and hepatic perfusion variables were evaluated. RESULTS: The median SF was higher in dogs < 3 years old (74.6%; n = 18) versus dogs ≥ 3 years old (35.1%; 18). Correlations were identified between SF and hepatic perfusion variables, and differences in results for SF and hepatic perfusion variables were detected between dogs grouped according to shunt type. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that CT-derived measurements of SF and hepatic perfusion variables in dogs with EPSS were feasible and could be useful (eg, estimating EPSS condition status and planning treatment) in clinical settings. In addition, our findings suggested that perfusion CT could be useful for distinguishing hemodynamic characteristics among different types of portosystemic shunts in dogs.

Inhibition of growth of canine-derived vascular endothelial cells by non-steroidal anti-inflammatory drugs and atrial natriuretic peptide.

This study investigated the direct effects of non-steroidal anti-inflammatory drugs (NSAIDs) and atrial natriuretic peptide (ANP) on canine-derived vascular endothelial cells (VECs). VECs were isolated and cultured from canine arteries and veins. The mRNA expressions of vascular endothelial growth factor receptor 2, cyclooxygenase-2, and natriuretic peptide receptor 1 were detected in the cultured VECs. The viability of the cultured VECs was reduced in a dose-dependent manner by meloxicam, carprofen, and robenacoxib. By contrast, dose escalations of ANP had only marginal influence on the viability of cultured VECs. NSAIDs may potentially serve as not only analgesic agents against cancerous and perioperative pain but also as adjuvant anti-angiogenic drugs in dogs with malignant tumors.

Surface imaging using a noncontact divergence-ratio axi-vision camera profilometer for laparoscopic and endoscopic applications.

Images from ordinary laparoscopes and endoscopes are two dimensional (2D), meaning the surgeon's depth perception is hindered. The proposed method supplements the 2D image with an image of the depth profile of the surface. The depth profile is obtained in real time without surface contact. The profilometer uses the same principle for acquiring distance information as the divergence-ratio axi-vision camera. The profilometer was added to an ordinary laparoscope with minimal increase in the weight and diameter of the shaft. With the profilometer added, there was a significant improvement in the ability to detect minute protrusions.

Omnifocus laparoscope.

The depth of focus of the latest-model "Chip-on-the-Tip" laparoscopes is limited to 10 cm. The proposed omnifocus laparoscope stretches this depth of focus to 160 cm. The proposed laparoscope is omnifocus, which means that all spots in the picture are in focus, not just certain designated spots as in autofocus devices. This is important because the entire scene needs to be focused during surgery. The omnifocus laparoscope is equipped with an array of color video cameras, each focused at a different distance. The distance information from the laparoscopic profilometer is used to generate a single omnifocused image.

Super deep 3D images from a 3D omnifocus video camera.

When using stereographic image pairs to create three-dimensional (3D) images, a deep depth of field in the original scene enhances the depth perception in the 3D image. The omnifocus video camera has no depth of field limitations and produces images that are in focus throughout. By installing an attachment on the omnifocus video camera, real-time super deep stereoscopic pairs of video images were obtained. The deeper depth of field creates a larger perspective image shift, which makes greater demands on the binocular fusion of human vision. A means of reducing the perspective shift without harming the depth of field was found.

Omnifocus video camera.

The omnifocus video camera takes videos, in which objects at different distances are all in focus in a single video display. The omnifocus video camera consists of an array of color video cameras combined with a unique distance mapping camera called the Divcam. The color video cameras are all aimed at the same scene, but each is focused at a different distance. The Divcam provides real-time distance information for every pixel in the scene. A pixel selection utility uses the distance information to select individual pixels from the multiple video outputs focused at different distances, in order to generate the final single video display that is everywhere in focus. This paper presents principle of operation, design consideration, detailed construction, and over all performance of the omnifocus video camera. The major emphasis of the paper is the proof of concept, but the prototype has been developed enough to demonstrate the superiority of this video camera over a conventional video camera. The resolution of the prototype is high, capturing even fine details such as fingerprints in the image. Just as the movie camera was a significant advance over the still camera, the omnifocus video camera represents a significant advance over all-focus cameras for still images.

Three-dimensional laparoscope based on the manipulation of polarized light by a cellophane half-wave plate.

A three-dimensional laparoscope that can capture three-dimensional images during surgery is reported. The principle is solely based on the manipulation of polarized light by a cellophane half-wave plate rather than computer processing; hence there is no time delay (it operates in real time) and lesions are viewed in true color, which is important for diagnostics. Three-dimensional images are obtained with a single laparoscope. A unique feature of this three-dimensional laparoscope is that it includes a virtual ruler to measure distances without physically touching the affected areas. The structure is simple, sturdy, lightweight, and its diameter is no bigger than a standard 10 mm diam laparoscope.

Three-dimensional camera phone.

An inexpensive technique for realizing a three-dimensional (3D) camera phone display is presented. Light from the liquid-crystal screen of a camera phone is linearly polarized, and its direction of polarization is easily manipulated by a cellophane sheet used as a half-waveplate. The novel 3D camera phone display is made possible solely by optical components without resorting to computation, so that the 3D image is displayed in real time. Quality of the original image is not sacrificed in the process of converting it into a 3D image.

High-definition real-time depth-mapping TV camera: HDTV Axi-Vision Camera.

We have developed a field-worthy, high-definition, real-time depth-mapping television camera called the HDTV Axi-Vision Camera. The camera can simultaneously capture both an ordinary HDTV color image and a depth image of objects on more than 1280 x 720 pixels at a frame rate of 29.97 Hz, or on 853 x 480 pixels at a frame rate of 59.94 Hz. The number of detectable pixels per unit time was increased by about 5 times that of the prototype camera by improving the sensitivity and resolution of the depthmapping camera. Short video clips demonstrate how depth information from the camera can be used to create a virtual image in actual television program production.

Gain-modulated Axi-Vision Camera (high speed high-accuracy depth-mapping camera).

We describe a novel real-time depth-mapping camera, the Gainmodulated Axi-Vision Camera, where pulsed laser light is combined with a gain-modulated camera. Depth resolution of 2.4 mm was obtained, which is higher than the resolution of the previously reported depth-mapping Axi- Vision Camera. Pixel-by-pixel depth information of 768 x 493 pixels is obtainable at one half of the video frame rate (15 Hz). A short movie clip is attached that illustrates the depth measurement operation. The merits of the Gain-modulated Axi-Vision Camera are high-resolution, real-time operation, and a relatively simple optical system. These merits primarily arise from the ultra-fast exposure time using a pulsed laser diode.