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Establishment of a new canine inflammatory mammary carcinoma cell line and analysis of its cystine-glutamate transporter subunit expression Full text
2022
Itoh, Harumichi | Naruse, Ryo | Tani, Kenji | Sunahara, Hiroshi | Nemoto, Yuki | Nakaichi, Munekazu | Iseri, Toshie | Horikirizono, Hiro | Itamoto, Kazuhito
Inflammatory mammary carcinoma (IMC) is a rare disease with a poor prognosis and one affecting dogs. Inflammatory breast carcinoma (IBC) is a subtype of malignant breast cancer in humans with a high degree of malignancy and a similarly poor prognosis. Since the clinical symptoms and prognoses of both are similar, canine IMC has been considered as a model of human IBC. In this study, we newly established a stable IMC-derived cell line from a patient at the Yamaguchi University Animal Medical Center in Japan. The patient was a female toy poodle presenting with an inflamed mammary gland, which was diagnosed as IMC. The cell line was established from a tissue biopsy. Surface antigen marker (CD24 and CD44) expression was determined. Cystine/glutamate antiporter (xCT) expression was determined by Western blotting, flow cytometry and fluorescence immunostaining, and sulfasalazine was administered to ascertain if it suppressed xCT expression. Stem cell marker (Nanog, Sox2, Myc and Klf4) expression and aldehyde dehydrogenase (ALDH) activity were also investigated. The cultured cells showed xCT, and its suppression showed downregulation of stem cell markers and ALDH activity. Stable cell proliferation was verified. A new canine IMC-derived cell line was established. In the future, we aim to study the effect of xCT on the maintenance of cancer stem cell properties in canine tumours, and propose a new therapeutic method for the treatment of canine IMC by targeting xCT.
Show more [+] Less [-]Cyclooxygenase-2 as a biomarker with diagnostic, therapeutic, prognostic, and predictive relevance in small animal oncology Full text
2020
Szweda, Marta | Rychlik, Andrzej | Babińska, Izabella | Pomianowski, Andrzej
In canine and feline populations, the number of neoplasm cases continues to increase around the world. Attempts are being made in centres of research to identify new biomarkers that speed up and improve the quality of oncological diagnostics and therapy in human and animal tumour patients. Cyclooxygenase-2 (COX-2) is a promising biomarker with increasing relevance to human oncology, but as yet with less application in veterinary oncology. The expression of COX-2 increases significantly during pathological processes involving inflammation, pain or fever. It is also overexpressed in humans presenting various types of tumours and in selected types of tumours in animals, particularly in dogs. This article discusses the expression of COX-2 in canine and feline tumours, the importance of COX-2 as a biomarker with diagnostic, therapeutic, prognostic and predictive relevance in oncology, and the clinical significance of inhibiting COX-2 overexpression in tumours.
Show more [+] Less [-]Retrospective analysis of indications and complications related to implantation of permanent pacemaker: 25 years of experience in 31 dogs Full text
2019
Noszczyk-Nowak, Agnieszka | Michałek, Marcin | Kapturska, Karolina | Cepiel, Alicja | Janiszewski, Adrian | Pasławski, Robert | Skrzypczak, Piotr | Pasławska, Urszula
Introduction: Pacemaker implantation is the only effective symptomatic treatment for life-threatening bradyarrhythmias. Major complications observed after implantation of cardiac pacemakers include lead dislocation, loss of pulse generator function, and inadequate stimulation. The aim of this retrospective single-centre study was to analyse the indications for pacemaker implantation and the incidence and types of complications associated with this procedure in dogs treated for symptomatic bradyarrhythmia. Material and Methods: The retrospective analysis included 31 dogs with symptomatic bradyarrhythmia, implanted with permanent cardiac pacemakers in 1992–2017. The list of analysed variables included patient age, breed, sex, indication for pacemaker implantation, comorbidities, and the incidence of procedure-related complications along with the type thereof. Results: The most common indication for pacemaker implantation was 3ʳᵈ degree AVB, followed by SSS, advanced 2ⁿᵈ degree AVB, and PAS. Pacemaker implantation was associated with a 35% overall complication rate and 6.45% periprocedural mortality. There were no significant differences in terms of procedure-related complications with regard to age, sex, breed, indications for pacemaker implantation, or comorbidities. Conclusions: Cardiac pacing is the only effective treatment of symptomatic bradycardia, but as an invasive procedure, may pose a risk of various complications, including death.
Show more [+] Less [-]Cloning and expression of NS3 gene of Pakistani isolate type 2 dengue virus Full text
2018
Yasmin, Farkhanda | Yaqub, Tahir | Idrees, Muhammad | Shahzad, Wasim | Hashmi, Abu Saeed | Aqil, Kiran | Mukhtar, Nadia | Zahoor, Muhammad Yasir | Akhtar, Naeem | Umar, Sajid
Dengue is one of the major emerging viral diseases in the world, with dramatic increases in reported cases in the last few decades and annual worldwide occurrence of approximately 390 million infections. It is a highly important mosquito-vectored disease and is a problem in tropical and subtropical areas of the world. The major aim of this study was to clone and express the dengue NS3 gene, in service to its therapeutic importance for the development of stable cell lines. Blood samples from dengue fever (DF) patients were collected and subjected to PCR amplification of the NS3 gene of dengue virus serotype-2 (DENV-2). The NS3 gene was amplified using gene specific primers and cloned in the TA cloning vectors. The gene was successfully expressed in mammalian expression vector pcDNA3.1. The current finding was different from a previously reported DENV-2 strain replicon constructed in different cells, in which the whole genetic material of the virus was used instead of an active protease gene, and which gave a low yield of replicon expressing cells. Recombinant NS3 could be used to produce an antibody that is possibly helpful for developing a single step diagnostic assay to detect the dengue virus NS3 antigen in sera of dengue patients.
Show more [+] Less [-]Characterisation of classical enterotoxins, virulence activity, and antibiotic susceptibility of Staphylococcus aureus isolated from Thai fermented pork sausages, clinical samples, and healthy carriers in northeastern Thailand Full text
2020
Sankomkai, Wanwisa | Boonyanugomol, Wongwarut | Kraisriwattana, Kairin | Nutchanon, Julalak | Boonsam, Kraisorn | Kaewbutra, Sasalux | Wongboot, Warawan
Contamination by Staphylococcus aureus of food produced from animal sources may have diverse and multifactorial causes that depend on geographical distribution. The goal of this study was to isolate and characterise S. aureus strains from contaminated fermented pork sausage, which is a local food of northeastern Thailand. S. aureus strains were isolated from local pork sausage, and the presence of classical enterotoxins was determined by PCR and reversed passive latex agglutination. These results were compared with strains derived from hospitalised patients and healthy carriers. Additionally, production of extracellular enzymes and haemolysin, biofilm formation, and antibiotic susceptibility were assessed. S. aureus was identified in 36 sausage isolates (60%). The strains positive for staphylococcal enterotoxin A were more frequently found in isolates from sausage and healthy carriers than in those from patients. All tested S. aureus strains were positive for DNase, lipase, proteinase, haemolysin, and biofilm formation; notably, strains isolated from food and healthy carriers displayed similar values. Most isolates were resistant to penicillin and ampicillin, while none were to methicillin. Thai fermented pork sausages are associated with a high risk of staphylococcal food poisoning, which may be linked to contamination caused by carriers. Dissemination of knowledge regarding best practices in sanitation and hygiene is important in local communities.
Show more [+] Less [-]Elastographic and morphological testicular changes in hypothyroidism – an experimental study Full text
2018
Gołyński, Marcin | Dębiak, Piotr | Gołyńska, Magdalena | Myśliwiec, Ewa | Szkodziak, Piotr | Kalisz, Grzegorz | Smiech, Anna | Lutnicki, Krzysztof | Szczepanik, Marcin
Methimazole-induced hypothyroidism is a clinical problem in the treatment of hyperthyroidism in people and animals and is an example of metabolic disease that can lead to fertility disorders and can give elastographic testicular changes. Ultrasound elastography using the Esaote MyLab Twice ultrasound system and a morphological examination of testes were performed in seven methimazole-administered (group E) and seven healthy rats (group C). The elasticity ratio of strains in the scrotal wall of the near-field test area to testicular tissue (ELX-T-RAT) and hardness percentage of strained tissues in the defined area of a testicle (ELX-T%HRD) in group E were statistically significantly lower than in group C. The degree of spermatogenesis was statistically significantly higher in group E than in group C and similarly seminiferous tubule diameters in group E were statistically significantly higher than in group C. Body weight and testicular weight in group E were statistically significantly lower than in group C. Changes in the elastographical parameters of testes may result from disorders secondary to hypothyroidism. The usefulness of elastography is noteworthy in the case of evaluation of testis function in patients with some metabolic disorders.
Show more [+] Less [-]C-kit, flt-3, PDGFR-β, and VEGFR2 expression in canine adrenal tumors and correlation with outcome following adrenalectomy Full text
2021
Harding, Kayla | De Mello Souza, Carlos H. | Shiomitsu, Keijiro | Maxwell, Elizabeth | Bertran, Judit
The objective of this retrospective study was to evaluate the expression of receptor tyrosine kinases (RTKs) in canine adrenal tumors and correlate this expression with features of tumor aggressiveness and survival in dogs undergoing adrenalectomy. Forty-three canine adrenal tumors were evaluated for expression of c-kit, fms-like tyrosine kinase 3 (flt-3), platelet-derived growth factor receptor-β (PDGFR-β), and vascular endothelial growth factor receptor 2 (VEGFR2) using immunohistochemistry. Tumor RTK staining characteristics were compared to normal adrenals. Medical records were reviewed for data regarding patient outcome and tumor characteristics. Expression of c-kit, flt-3, PDGFR-β, and VEGFR2 was detected in 26.9%, 92.3%, 96.2%, and 61.5% of cortical tumors and 0%, 63.2%, 47.4%, and 15.8% of pheochromocytomas, respectively. Expression of RTKs was not significantly increased when compared to normal adrenals and did not correlate with survival after adrenalectomy. Receptor tyrosine kinases are not overexpressed in canine adrenal tumors compared to normal adrenal tissue. Therapeutic inhibition of these receptors may still represent an effective approach in cases where receptor activation is present.
Show more [+] Less [-]Computed tomographic evaluation of pancreatic perfusion in healthy dogs Full text
2020
Kloer, Timothy B. | Rao, Sangeeta | Twedt, David C. | Marolf, Angela J.
OBJECTIVE To evaluate the feasibility of contrast-enhanced CT for assessment of pancreatic perfusion in healthy dogs. ANIMALS 6 healthy purpose-bred female Treeing Walker Coonhounds. PROCEDURES Contrast-enhanced CT of the cranial part of the abdomen was performed with 3-mm slice thickness. Postprocessing computer software designed for evaluation of human patients was used to calculate perfusion data for the pancreas and liver by use of 3-mm and reformatted 6-mm slices. Differences in perfusion variables between the pancreas and liver and differences in liver-specific data of interest were evaluated with the Friedman test. RESULTS Multiple pancreatic perfusion variables were determined, including perfusion, peak enhancement index, time to peak enhancement, and blood volume. The same variables as well as arterial, portal, and total perfusion and hepatic perfusion index were determined for the liver. Values for 6-mm slices appeared similar to those for 3-mm slices. The liver had significantly greater median perfusion and peak enhancement index, compared with the pancreas. CONCLUSIONS AND CLINICAL RELEVANCE Measurement of pancreatic perfusion with contrast-enhanced CT was feasible in this group of dogs. Hepatic arterial and pancreatic perfusion values were similar to previously published findings for dogs, but hepatic portal and hepatic total perfusion measurements were not. These discrepancies might have been attributable to physiologic differences between dogs and people and related limitations of the CT software intended for evaluation of human patients. Further research is warranted to assess reliability of perfusion variables and applicability of the method for assessment of canine patients with pancreatic abnormalities.
Show more [+] Less [-]Correlation of Activity Data in Normal Dogs to Distance Traveled Full text
2020
Eskander, Bishoy S. | Barbar, Megan | Evans, Richard B. | Enomoto, Mastaka | Lascelles, B. Duncan X. | Conzemius, Michael G.
The objective of this study was to explore the mathematical relationships between independent variables (patient morphometrics and treadmill speed) and dependent variables (accelerometer or pedometer output) when evaluating data from accelerometers and pedometers in dogs. Twenty dogs took part in 3 randomized activities, consisting of exercise on a treadmill at 1.0, 1.5, and 2.0 m/s for a total distance of 1 km at each speed. Dogs simultaneously wore both an accelerometer and a pedometer. Statistical analysis used multiple regression models to discover the relationships between independent and dependent variables. A formula was developed to predict the distance traveled by a dog based on its morphometrics and activity monitor output. Shoulder height had stronger correlations to accelerometer and pedometer outputs than other morphometric variables. As shoulder height increased, all accelerometer and pedometer outputs decreased. As treadmill speed increased, both accelerometer and pedometer step counts decreased, while accelerometer activity counts increased. According to a formula derived to predict the total distance traveled using patient shoulder height and accelerometer or pedometer output, pedometer steps were the most accurate predictor of distance traveled. Accelerometer steps were less accurate when using the same model. Accelerometer activity counts did not reveal a meaningful predictive formula. The results of this study indicate that patient morphometrics and treadmill speed (as a measure of intensity) influenced accelerometer and pedometer data. The pedometer data more precisely and accurately estimated the distance traveled based on step counts and patient shoulder height. In normal dogs, accelerometer and pedometer steps may reasonably estimate distance traveled.
Show more [+] Less [-]Use of three-dimensional time-of-flight magnetic resonance angiography at 1.5 Tesla to evaluate the intracranial arteries of 39 dogs with idiopathic epilepsy Full text
2019
Ishikawa, Chieko | Ito, Daisuke | Tanaka, Natsumi | Kitagawa, Masato
OBJECTIVE To assess visualization of the intracranial arteries and internal carotid artery (ICA) on 3-D time-of-flight (TOF) magnetic resonance angiography (MRA) images obtained at 1.5 T and to investigate factors that affect the image quality of those arteries in dogs. ANIMALS 39 dogs with idiopathic epilepsy. PROCEDURES Each dog underwent 3-D TOF MRA, and 5 pairs of intracranial arteries, the basilar artery, and both ICAs were evaluated. Each artery was assigned an image-quality score on a scale of 0 to 3, where 0 = poor and 3 = excellent. Multivariable regression analysis was used to assess whether age, body weight (BW), serum total cholesterol concentration, intracranial volume (ICV), and mean arterial pressure were significantly associated with the image quality of each vessel. RESULTS In all dogs, the image-quality score was 2 or 3 for the proximal middle cerebral arteries, basilar artery, and caudal aspect of the caudal communicating arteries. In some dogs, the rostral cerebellar arteries, rostral aspect of the caudal communicating arteries, and middle and rostral aspects of the ICA were poorly visualized. For various arteries, image quality was negatively associated with age and positively associated with BW and ICV. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that 3-D TOF MRA images obtained at 1.5 T did not consistently and clearly delineate the ICA and narrow or peripheral intracranial arteries of dogs; therefore, careful attention is required when such images are assessed. Patient age, BW, and ICV can also affect the image quality of some intracranial arteries on 3-D TOF MRA images.
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