Impact of Adipose Tissue Depot Harvesting Site on the Multilineage Induction Capacity of Male Rat Adipose-Derived Mesenchymal Stem Cells: An In Vitro Study.

Recently, substantial attention has been paid toward adipose-derived mesenchymal stem cells (AdMSCs) as a potential therapy in tissue engineering and regenerative medicine applications. Rat AdMSCs (r-AdMSCs) are frequently utilized. However, the influence of the adipose depot site on the multilineage differentiation potential of the r-AdMSCs is still ambiguous. Hence, the main objective of this study was to explore the influence of the adipose tissue harvesting location on the ability of r-AdMSCs to express the stem-cell-related markers and pluripotency genes, as well as their differentiation capacity, for the first time. Herein, we have isolated r-AdMSCs from the inguinal, epididymal, peri-renal, and back subcutaneous fats. Cells were compared in terms of their phenotype, immunophenotype, and expression of pluripotency genes using RT-PCR. Additionally, we investigated their potential for multilineage (adipogenic, osteogenic, and chondrogenic) induction using special stains confirmed by the expression of the related genes using RT-qPCR. All cells could positively express stem cell marker CD 90 and CD 105 with no significant in-between differences. However, they did not express the hematopoietic markers as CD 34 and CD 45. All cells could be induced successfully. However, epididymal and inguinal cells presented the highest capacity for adipogenic and osteogenic differentiation (21.36-fold and 11.63-fold for OPN, 29.69-fold and 26.68-fold for BMP2, and 37.67-fold and 22.35-fold for BSP, respectively, in epididymal and inguinal cells (p < 0.0001)). On the contrary, the subcutaneous cells exhibited a superior potential for chondrogenesis over the other sites (8.9-fold for CHM1 and 5.93-fold for ACAN, (p < 0.0001)). In conclusion, the adipose tissue harvesting site could influence the differentiation capacity of the isolated AdMSCs. To enhance the results of their employment in various regenerative cell-based therapies, it is thus vital to take the collection site selection into consideration.

Diurnal rhythms in testicular blood flow, testicular morphometry and reproductive hormones in Shiba goats.

CONTEXT: Testicular blood flow (TBF) is crucial for testicular function. The pattern of TBF in Shiba goats indicates seasonal variations. AIMS: This study aimed to investigate the effect of diurnal variations on TBF, testis volume (TV), testicular echogenicity, and reproductive hormones in goats over a 24-h period. METHODS: In three trials that went for three consecutive days each, 12 bucks were scanned using Triplex ultrasonography to assess the TV, pixel intensity of testicular echotexture (PIX), and Doppler indices of TBF (resistive index: RI and pulsatility index: PI) in four-time points a day (at 6.00, 12.00, 18.00, and 00.00h). Concomitantly, the changes in circulating FSH, LH, inhibin, testosterone (T), estradiol (E2), cortisol, and melatonin were assessed. KEY RESULTS: Results revealed diurnal alterations in the calculated RI of TBF and the PIX of testicular parenchyma (P <0.05). Lower RI values of the TBF were observed at 6.00h compared to other time points. There were significant diurnal alterations in the levels of FSH (P <0.05), LH (P <0.05), T (P <0.0001), E2 (P <0.0001), cortisol (P <0.0001), and melatonin (P <0.0001). FSH attained a higher concentration at 18.00h compared to 12.00h. Concentrations of LH were significantly higher at 06.00h compared to those at 18.00h. Concentrations of T were significantly higher at 6.00 compared to other time points. E2 showed higher concentrations at 6.00h and 00.00h compared to 12.00h and 18.00h. On the contrary, concentrations of cortisol were significantly higher at 12.00h and 18.00h compared to 06.00h and 00.00h. The highest concentrations of melatonin were observed at 00.00h compared to other time points, while the lowest concentrations were at 12.00h. CONCLUSIONS: Diurnal rhythm induces significant changes in TBF, testicular PIX, and circulating FSH, LH, T, E2, cortisol, and melatonin over the 24-hday. IMPLICATIONS: The outcomes of the study are reflected in the advisability of monitoring the TBF at a fixed time a day to avoid the circadian rhythm effect.

Adjunct ambrisentan therapy had clinical benefits in 5 dogs with sildenafil-refractory pulmonary hypertension.

Although sildenafil is used in dogs with severe pulmonary hypertension, they sometimes become resistant and clinical signs deteriorate over time. The objective of this study was to determine the benefits of adjunct ambrisentan therapy in dogs with sildenafil-refractory pulmonary hypertension. In 5 dogs with severe pulmonary hypertension with deteriorating clinical signs despite ongoing sildenafil treatment, adding ambrisentan improved appetite, activity, and respiratory functions. Although peak tricuspid valve regurgitation velocity, as measured by Doppler echocardiography, did not necessarily decrease after ambrisentan administration, there was improved partial pressure of arterial oxygen and the alveolar-arterial oxygen gradient, with no apparent side effects. We concluded that ambrisentan has potential as an adjunct treatment in dogs with pulmonary hypertension that are refractory to sildenafil therapy. Key clinical message: Ambrisentan improved clinical signs in dogs with sildenafil-refractory pulmonary hypertension.

Le traitement d'appoint à l'ambrisentan a eu des avantages cliniques chez cinq chiens atteints d'hypertension pulmonaire réfractaire au sildénafil. Bien que le sildénafil soit utilisé chez les chiens souffrant d'hypertension pulmonaire sévère, ils deviennent parfois résistants et les signes cliniques s'aggravent avec le temps. L'objectif de cette étude était de déterminer les avantages d'un traitement d'appoint à l'ambrisentan chez les chiens souffrant d'hypertension pulmonaire réfractaire au sildénafil. Chez cinq chiens souffrant d'hypertension pulmonaire sévère avec détérioration des signes cliniques malgré un traitement continu au sildénafil, l'ajout d'ambrisentan a amélioré l'appétit, l'activité et les fonctions respiratoires. Bien que la vitesse maximale de régurgitation de la valve tricuspide, mesurée par échocardiographie Doppler, n'ait pas nécessairement diminué après l'administration d'ambrisentan, la pression partielle d'oxygène artériel et le gradient alvéolo-artériel d'oxygène ont été améliorés, sans effets secondaires apparents. Nous avons conclu que l'ambrisentan a un potentiel en tant que traitement d'appoint chez les chiens souffrant d'hypertension pulmonaire qui sont réfractaires au traitement par le sildénafil.Message clinique clé:L'ambrisentan a amélioré les signes cliniques chez les chiens souffrant d'hypertension pulmonaire réfractaire au sildénafil.(Traduit par Dr Serge Messier).

Successful ligation of the left and right gastric vein in a dog with congenital portosystemic shunts type Aii.

The type Aii shunt is a congenital extrahepatic portosystemic shunt (ePSS) involving the left and right gastric vein and the caudal vena cava (CVC). This report describes the case of a 6-month-old Italian greyhound diagnosed with a type Aii large-diameter ePSS. Staged surgeries were employed to completely ligate the 2 gastric veins and to avoid the risk of traumatizing the shunt vessel, CVC, and celiac artery. Clinical signs improved postoperatively, and after 3 years, ultrasonography demonstrated no evidence of reoccurrence. This procedure provides an alternative surgical option for correction of ePSS type Aii. Key clinical message: This case report demonstrates congenital PSS involving the left and right gastric vein and the caudal vena could be treated with both ligation of left and right gastric vein. This technique could decrease the risk of traumatizing the shunt vessel, CVC, and celiac artery.

Ligature réussie des veines gastriques gauche et droite chez un chien avec des shunts portosystémiques congénitaux de type Aii. Le shunt de type Aii est un shunt porto-systémique extrahépatique congénital (ePSS) impliquant la veine gastrique gauche et droite et la veine cave caudale (CVC). Ce rapport décrit le cas d'un lévrier italien de 6 mois diagnostiqué avec un ePSS de grand diamètre de type Aii. Des chirurgies par étapes ont été effectuées pour ligaturer complètement les deux veines gastriques et pour éviter le risque de traumatiser le vaisseau avec shunt, la CVC et l'artère coeliaque. Les signes cliniques se sont améliorés après l'opération et après trois ans, l'échographie n'a montré aucun signe de récidive. Cette procédure offre une option chirurgicale alternative pour la correction de l'ePSS de type Aii.Message clinique clé :Ce rapport de cas démontre un ePSS congénital impliquant la veine gastrique gauche et droite et la veine caudale pouvant être traité par la ligature de la veine gastrique gauche et droite. Cette technique pourrait diminuer le risque de traumatiser le vaisseau avec shunt, le CVC et l'artère coeliaque.(Traduit par Dr Serge Messier).

Autologous tunica vaginalis communis flap for repairing an abdominal wall hernia in a dog.

An 8-year-old, intact male miniature dachshund dog, weighing 8.6 kg, was presented with a soft swelling in the caudal abdominal region, including both sides of the groin area. Laparotomy revealed a severe caudal abdominal wall hernia with atrophy of the rectus abdominal muscle. The defect was repaired using a tunica vaginalis communis flap following a standard open prescrotal castration. There were no complications or recurrence of the hernia at 11 months after surgery. This surgical technique involves autogenous reconstruction, is easy to perform, and requires minimal dissection. The tunica vaginalis communis flap has potential clinical applications for repairing caudal abdominal wall hernias in male dogs.

Lambeau de tunique vaginale autologue pour la réparation d'une hernie de la paroi abdominale chez un chien. Un teckel miniature mâle intact de 8 ans, pesant 8,6 kg, a présenté une enflure molle dans la région abdominale caudale, y compris des deux côtés de la région de l'aine. La laparotomie a révélé une hernie sévère de la paroi abdominale caudale avec atrophie du muscle abdominal droit. Le défaut a été réparé à l'aide d'un lambeau de la tunique vaginale après une castration préscrotale ouverte standard. Il n'y a eu aucune complication ni récidive de la hernie 11 mois après la chirurgie. Cette technique chirurgicale implique une reconstruction autogène, est facile à réaliser et nécessite une dissection minimale. Le lambeau de tunique vaginale a des applications cliniques potentielles pour la réparation des hernies de la paroi abdominale caudale chez les chiens mâles.(Traduit par Dr Serge Messier).

Changes of inflammatory and oxidative stress biomarkers in dogs with different stages of heart failure.

BACKGROUND: Heart failure (HF) is associated with changes in inflammatory and oxidative stress biomarkers. This study aimed to evaluate the changes of a panel of inflammatory and oxidative stress biomarkers in dogs with different stages of HF and its relation with the severity of the disease and echocardiographic changes. A total of 29 dogs with HF as a result of myxomatous mitral valve degeneration or dilated cardiomyopathy were included and classified as stage-A (healthy), B (asymptomatic dogs), C (symptomatic dogs) and D (dogs with end-stage HF) according to the ACVIM staging system. In these dogs an ecnhocardiographic examination was performed and cytokines, and inflammatory and oxidative stress markers were evaluated in serum. RESULTS: KC-like was significantly increased in dogs of stage-C (P < 0.01) and -D (P < 0.05) compared with stage-A and -B. Stage-D dogs showed significantly higher serum CRP and Hp (P < 0.05) but lower serum antioxidant capacity (PON1, TEAC, CUPRAC, and thiol) compared to stage-A and -B (P < 0.05). After the treatment, serum levels of CRP, Hp and KC-like decreased and serum antioxidant levels increased compared to their pre-treatment values. Left ventricular dimension and LA/Ao ratio correlated positively with CRP, MCP-1, and KC-like but negatively with PON1, GM-CSF, IL-7 and antioxidant biomarkers (P < 0.01). CONCLUSION: Our results showed that dogs with advanced HF show increases in positive acute-phase proteins and selected inflammatory cytokines such as KC-like, and decreases in antioxidant biomarkers, indicating that inflammation and oxidative stress act as collaborative partners in the pathogenesis of HF. Some of these biomarkers of inflammation and oxidative stress could have the potential to be biomarkers to monitor the severity of the disease and the effect of treatment.

Platelet proteome changes in dogs with congestive heart failure.

BACKGROUND: Platelets play a central role in the development of cardiovascular diseases and changes in their proteins are involved in the pathophysiology of heart diseases in humans. There is lack of knowledge about the possible role of platelets in congestive heart failure (CHF) in dogs. Thus, this study aimed to investigate the changes in global platelet proteomes in dogs with CHF, to clarify the possible role of platelets in the physiopathology of this disease. Healthy-dogs (n = 10) and dogs with acute CHF due to myxomatous mitral valve disease (MMVD, n = 10) were used. Acute CHF was defined based on the clinical (increased respiratory rate or difficulty breathing) and radiographic findings of pulmonary edema. Dogs Blood samples were collected into tubes with acid-citrate-dextrose, and platelet-pellets were obtained by centrifuge and washing steps. Platelet-proteomes were identified using LC-MS based label-free differential proteome expression analysis method and matched according to protein database for Canis lupus familiaris. RESULTS: Totally 104 different proteins were identified in the platelets of the dogs being 4 out of them were significantly up-regulated and 6 down-regulated in acute CHF dogs. Guanine-nucleotide-binding protein, apolipoproteins (A-II and C-III) and clusterin levels increased, but CXC-motif-chemokine-10, cytochrome-C-oxidase-subunit-2, cathepsin-D, serine/threonine-protein-phosphatase-PP1-gamma-catalytic-subunit, creatine-kinase-B-type and myotrophin levels decreased in acute CHF dogs. These proteins are associated with several molecular functions, biological processes, signaling systems and immune-inflammatory responses. CONCLUSION: This study describes by first time the changes in the protein composition in platelets of dogs with acute CHF due to MMVD. Our findings provide a resource for increase the knowledge about the proteome of canine platelets and their roles in CHF caused by MMVD and could be a tool for further investigations about the prevention and treatment of this disease.

Metabolic and chemical regulation of tRNA modification associated with taurine deficiency and human disease.

Modified uridine containing taurine, 5-taurinomethyluridine (τm5U), is found at the anticodon first position of mitochondrial (mt-)transfer RNAs (tRNAs). Previously, we reported that τm5U is absent in mt-tRNAs with pathogenic mutations associated with mitochondrial diseases. However, biogenesis and physiological role of τm5U remained elusive. Here, we elucidated τm5U biogenesis by confirming that 5,10-methylene-tetrahydrofolate and taurine are metabolic substrates for τm5U formation catalyzed by MTO1 and GTPBP3. GTPBP3-knockout cells exhibited respiratory defects and reduced mitochondrial translation. Very little τm5U34 was detected in patient's cells with the GTPBP3 mutation, demonstrating that lack of τm5U results in pathological consequences. Taurine starvation resulted in downregulation of τm5U frequency in cultured cells and animal tissues (cat liver and flatfish). Strikingly, 5-carboxymethylaminomethyluridine (cmnm5U), in which the taurine moiety of τm5U is replaced with glycine, was detected in mt-tRNAs from taurine-depleted cells. These results indicate that tRNA modifications are dynamically regulated via sensing of intracellular metabolites under physiological condition.

Left ventricular vortex and intraventricular pressure difference in dogs under various loading conditions.

Restrictions on the conventional evaluation of diastolic function have been recognized, especially under various loading conditions. Recently, new noninvasive ventricular vortex indexes have been introduced and are expected to reflect the cardiac function. Physiologically, there is a hypothesis that the intraventricular pressure difference (IVPD) is related to the formation of vortexes. IVPD and vortex indexes were simultaneously measured, and the relationship between the two was investigated. To verify the possibility of diastolic vorticity as an index of diastolic relaxation, a correlation between diastolic vorticity and the load dependency of vorticity [time constant (τ)] was examined. Six healthy dogs were studied using transthoracic echocardiography, pressure, and a conductance catheter. Vorticity was analyzed using vector flow mapping (VFM). IVPD was determined using Euler's equation with color M-mode Doppler images. Data were obtained at baseline, at balloon dilatation in the thoracic aorta to alter afterload, at hydroxyethyl starch infusion to alter preload, and at milrinone administration to alter ventricular relaxation. Peak vorticity at early diastole (E-Vor) and IVPD of the midventricle (MIVPD) decreased under pressure loading, were unchanged under volume loading, and increased during milrinone administration. In multivariate analysis, the independent predictors of τ were global longitudinal strain, strain rate at early diastole, and E-Vor. MIVPD was strongly correlated with E-Vor ( r = 0.84). VFM-derived peak E vorticity was strongly related to IVPD, especially MIVPD, under various loading conditions. Both of these novel indexes are promising as reliable indexes of ventricular relaxation, independent from preload. NEW & NOTEWORTHY We showed the close relationship of vortex and intraventricular pressure difference and showed that both of them can become new markers of the left ventricular relaxation property. Our present study creates a paradigm for future studies in the field of intraventircular flow physiology and clinical diastology.

Contrasting Effects of Inhibition of Phosphodiesterase 3 and 5 on Cardiac Function and Interstitial Fibrosis in Rats With Isoproterenol-Induced Cardiac Dysfunction.

Myocardial relaxation and stiffness are influenced by fibrillar collagen content. Cyclic nucleotide signaling regulators have been investigated targeting more effective modulation of collagen deposition during myocardial healing process. To assess the effects of phosphodiesterase type 3 and phosphodiesterase type 5 inhibitors on cardiac function and left ventricular myocardial fibrosis in catecholamine-induced myocardial injury, sildenafil and pimobendan were administered to male Wistar rats 24 hours after isoproterenol injection. Echocardiography and electrocardiogram were performed to assess kinetic and rhythm changes during 45 days of drug administration. At the end of study, type I and type III collagen were measured through immunohistochemistry analysis, and left ventricular pressure was assessed through invasive method. Echocardiography assessment showed increased relative wall thickness at 45 days in pimobendan group with significant diastolic dysfunction and increased collagen I deposition compared with nontreated positive group (3.03 ± 0.31 vs. 2.73 ± 0.28%, P < 0.05). Diastolic pressure correlated positively with type I collagen (r = 0.54, P < 0.05). Type III collagen analysis did not demonstrate difference among the groups. Sildenafil administration attenuated type I collagen deposition (2.15 ± 0.51 vs. positive group, P < 0.05) and suggested to be related to arrhythmic events. Arrhythmic events were not related to the quantity of fibrillar collagen deposition. Although negative modulation of collagen synthesis through cyclic nucleotides signaling have shown promising results, in this study, pimobendan postconditioning resulted in increased collagen type I formation and severe diastolic dysfunction while sildenafil postconditioning reduced collagen type I deposition and attenuated diastolic dysfunction.

Inotropic and lusitropic effects of incremental doses of dobutamine in dogs with right ventricular apical pacing-induced cardiac dysfunction.

This study aimed to assess the effects of incremental doses of dobutamine on diastolic function in healthy and rapid ventricular apical pacing (RVAP)-induced cardiac dysfunction anesthetized dogs. Inotropic and lusitropic effects of dobutamine (2, 4, 8, and 12 µg kg-1  min-1 ) were assessed through left ventricle (LV) pressure-volume relation and Doppler echocardiography in six female dogs before and after 8 weeks of RVAP. Peak rate of LV pressure fall (-dP/dtmin ) improved with doses >4 µg kg-1  min-1 in healthy (4,490 ± 970 vs. 3,265 ± 471 mmHg/s, p < 0.05) and >8 µg kg-1  min-1 in RVAP dogs (3,385 ± 1,122 vs. 1,864 ± 849 mmHg/s, p < 0.05) while the time constant of relaxation (tau) reduced with doses >4 µg kg-1  min-1 in both groups (healthy: 24.0 ± 3.7 vs. 28.2 ± 4.9 ms; RVAP: 32.6 ± 8.5 vs. 37.5 ± 11.4 ms, p < 0.05) comparing with baseline. Indices of relaxation (-dP/dtmin and tau) suggested preserved lusitropic response in contrast with markedly reduced indices of contractility in the RVAP group compared with healthy group at same infusion rates. Doppler echocardiography showed significant reduction of elastic recoil in failing hearts. The results of this study demonstrated maximal positive lusitropic effects of dobutamine at a dose of 8 µg kg-1  min-1 in ventricular pacing-induced cardiac dysfunction without further impairment of ventricular filling.

Assessment of the subcutaneous degradation process of insoluble hyaluronic acid in rats.

Insoluble hyaluronic acid (IHA) may prevent adhesions by forming a physical barrier during the period when postoperative adhesions form. This study was performed to verify the changes that a solid IHA membrane undergoes as it is degraded in vivo, and to ascertain the swelling rate of IHA required for it to function as a physical barrier during the postoperative adhesion formation period. Nine female WI rats weighing 300-400 g were used. Discs 8 mm in diameter were cut out of dry IHA membranes made of IHA with a swelling rate (wet weight/dry weight) of either 2.47 (high-swelling IHA) or 1.94 (low-swelling IHA). They were placed in saline to swell and then washed with saline before subcutaneous implantation in four pockets in each rat. The high-swelling IHA started to degrade more rapidly than the low-swelling IHA. There was no evidence of degradation of the low-swelling IHA until day 7, but once it had started, the speed of degradation tended to be similar to that of the high-swelling IHA. The present results showed that, when IHA is implanted subcutaneously in rats, it is degraded over time in a phased process. The swelling rate required for the use of IHA as a postoperative adhesion barrier was also suggested.

Development of an Anti-Adhesive Membrane for Use in Video-Assisted Thoracic Surgery.

Background: The need to prevent postoperative adhesions after surgery has been considered a significant challenge in thoracic surgery, especially with the advent of video-assisted thoracic surgery (VATS). While preventive materials for postoperative adhesions have been studied for many years, they are all still in the development phases. Methods: In this animal study, an insoluble hyaluronic acid membrane was used in VATS for wedge resection to test its operability and to examine the body's response to the membrane. Ten beagles were divided into two groups, an experimental group and a negative control group. In the experimental group, an insoluble hyaluronic acid membrane containing glycerol was used as the test membrane (10 x 10 x 0.1 cm3). The test membrane was implanted in the left thoracic cavity of the animal under VATS following wedge resection. The animals were observed for two weeks and then euthanized for examination. Results: Macroscopically, the median adhesion score was lower in the experimental group (0) than in the control group (2.5). On histopathological examination, the test membrane elicited only a minor inflammatory response and foreign body reaction. Conclusion: The test membrane showed satisfactory operability and appears to be a practical material to prevent postoperative adhesions after thoracic surgery in VATS.

The relationship between systolic vector flow mapping parameters and left ventricular cardiac function in healthy dogs.

Vector flow mapping (VFM) is a novel echocardiographic technology that shows blood flow vectors and vortexes, enabled the hydrokinetic evaluation of hemodynamics within the left ventricle. VFM provides several unique parameters: circulation, vorticity, vortex area, and energy loss. The present study aims to reveal a relationship between VFM parameters and cardiac function. Five healthy Beagle dogs were anesthetized and administered with dobutamine (0, 2, 4, 8, 12 µg/kg/min). Pressure-volume diagrams were acquired to assess cardiac function using pressure-volume conductance catheter. Systolic maximum circulation, vorticity, vortex area, and energy loss were measured using VFM. The systolic maximum circulation, systolic vorticity, systolic vortex area, and systolic energy loss were increased by dobutamine administration. There was a strongly significant correlation between the systolic maximum circulation and ejection fraction (r = 0.76), maximal positive left ventricular (LV) pressure derivatives (dP/dt max) (r = 0.80), and end-systolic LV elastance (r = 0.73). Systolic vorticity and systolic vortex area were strongly correlated with ejection fraction (r = 0.76, 0.68) and dP/dt max (r = 0.76, 0.69), and end-systolic LV elastance (r = 0.62, 0.74), respectively. Systolic energy loss was strongly correlated with dP/dt max (r = 0.78), systolic maximum circulation (r = 0.81), and systolic vorticity (r = 0.82). The present study revealed that systolic VFM parameters are associated with the LV contractility. Furthermore, systolic energy loss was susceptible to the systolic vortex parameters such as systolic vorticity and systolic maximum circulation. Systolic VFM parameters are new hydrokinetic indices reflecting LV contractility.

Utility of Lung Ultrasonography for Detection of Pleural Adhesions in Dogs.

OBJECTIVES: To assess lung respiratory movement ("lung sliding") in dogs using B-mode ultrasonography (US) and to develop a method that assesses adhesions between the parietal pleura and the lung. METHODS: Seventeen male beagles were anesthetized, and respiratory management was performed with intermittent positive pressure ventilation. Lung-sliding assessments and adhesion examinations were performed with lung US under general anesthesia before and 2 weeks after thoracotomy. Lung sliding was scored on a 4-level scale based on the percentage of the area that showed lung sliding (3, an area of roughly ≥80% of the intercostal space; 2, about 50% of the area of the intercostal space; 1, a small area of the intercostal space; or 0, movement absent); scores of 0, 1, and 2 indicated adhesions, whereas a score of 3 indicated no adhesions. The animals were then euthanized, and necropsy was performed to examine pleural adhesions. RESULTS: Lung US and necropsy findings were compared. The median lung-sliding score for the 12 sites with pleural adhesions on necropsy was 1.5, whereas it was 3.0 for the 532 sites without pleural adhesions. The lung-sliding score was significantly lower in the group with adhesions (P < .0001). Adhesion sites detected on necropsy were in accordance with the sites that had decreased lung-sliding scores. Lung US could detect pleural adhesions with sensitivity of 100.0% and specificity of 87.8%. CONCLUSIONS: Examination of lung sliding by thoracic US has high diagnostic value for detecting canine pleural adhesions and is useful in predicting adhesion sites before thoracic surgery in healthy dogs.

Development of a new surgical sheet containing both silk fibroin and thermoplastic polyurethane for cardiovascular surgery.

PURPOSE: The surgical sheets that are currently used for congenital cardiovascular surgery have several drawbacks, including material deterioration, calcification, and pseudo-intimal proliferation resulting in hemodynamic disturbance. The aim of this study was to evaluate a newly developed sheet made from a combination of silk fibroin (SF) and a synthetic polymer, thermoplastic polyurethane (TPU), for surgical use. METHODS: The hybrid SF/TPU sheet was a non-woven fabric with nanofibers that was made using the electrospinning method. The mechanical properties of the SF/TPU sheet were characterized. To determine its biocompatibility, part of the wall of the canine descending aorta was replaced with a SF/TPU sheet as a patch. The patches were removed after 3 months and a histological examination was performed. RESULTS: The flexibility, water permeability, and suture retention strength of the SF/TPU sheet were excellent and equivalent to those of existing sheets. The SF/TPU sheet had excellent handling properties and fit well into the vascular wall without needle hole bleeding. The histological examination revealed that the intimal tissue was restored well over the intraluminal surface of the explanted SF/TPU sheet, the absence of calcium deposition, and minimal inflammatory reaction, without signs of degradation. CONCLUSION: The SF/TPU sheet had excellent mechanical properties and tissue biocompatibility. These favorable features and possible biodegradability of the SF portion warrant a long-term follow-up study.

Dose-dependent effects of isoflurane and dobutamine on cardiovascular function in dogs with experimental mitral regurgitation.

OBJECTIVE: To investigate the dose-dependent effects of isoflurane and dobutamine on haemodynamics in dogs with experimentally induced mitral valve insufficiency (MI). STUDY DESIGN: Experimental, dose-response study. ANIMALS: Six healthy Beagle dogs. METHODS: Dogs with surgically induced MI were anaesthetized once. First, anaesthesia was maintained at an end-tidal isoflurane concentration (Fe'Iso) 1.0% (ISO1.0) for 20 minutes. Then, dobutamine was infused successively at 2, 4, 8 and 12 µg kg-1 minute-1 (DOB2-12) for 10 minutes at each dose rate. Measurements were recorded at each stage. Dobutamine was discontinued and Fe'Iso was increased to 1.5% (ISO1.5) for 20 minutes. Dobutamine was administered similarly to ISO1.0, and cardiovascular variables were recorded. The same sequence was repeated for Fe'Iso 2.0% (ISO2.0). Aortic pressure (AoP) and left atrial pressure (LAP) were recorded by radiotelemetry. The combination method of the pressure-volume loop analysis and transoesophageal echocardiography was used to measure cardiovascular variables: end-systolic elastance (Ees), effective arterial elastance (Ea), Ea/Ees, forward stroke volume (FSV), heart rate (HR), and cardiac output (CO). RESULTS: High isoflurane concentration resulted in reduced Ees and increased Ea/Ees, which indicated low arterial pressure. High-dose dobutamine administration resulted in increased Ees and FSV at all isoflurane concentrations. In ISO1.5 and ISO2.0, HR was lower at DOB4 than baseline (BL) but increased at DOB12 compared with DOB4. CO increased at ≥ DOB8 compared with BL. In ISO1.5 and ISO2.0, systolic and mean AoP increased at ≥ DOB4 and ≥ DOB8, respectively. LAP did not change under all conditions. CONCLUSIONS AND CLINICAL RELEVANCE: The dose-dependent hypotensive effect of isoflurane in MI dogs was mainly derived from the decrease in contractility. Dobutamine increased AoP without increasing LAP by increasing the contractility attenuated by isoflurane. Our findings may improve the cardiovascular management of dogs with MI undergoing general anaesthesia with isoflurane.

Hybrid surgical treatment for 2 feline cases of intrahepatic shunt.

Intrahepatic portosystemic shunt was encountered in 2 cats (10 and 5 months old) exhibiting neurological symptoms and general deterioration. Both cats were treated with coil embolization using a hybrid surgical technique combining conventional open surgery and interventional radiology techniques, achieving good postoperative outcomes (follow-up: 22 and 10 months, respectively).

Traitement chirurgical hybride pour deux cas félins de shunt intrahépatique. Un shunt portosystémique intrahépatique a été observé chez deux chats (âgés de 10 et de 5 mois) manifestant des symptômes neurologiques et une détérioration générale. Les deux chats ont été traités par une embolisation utilisant des spires à l'aide d'une technique chirurgicale hybride combinant une chirurgie ouverte conventionnelle et des techniques de radiologie d'intervention afin d'obtenir de bons résultats postopératoires (suivi : 22 et 10 mois, respectivement).(Traduit par Isabelle Vallières).

Evaluation of endothelialization in the center part of graft using 3 cm vascular grafts implanted in the abdominal aortae of the rat.

In order to develop small-diameter vascular grafts, it is necessary to evaluate endothelialization, especially, in the center part at early stage. For implantation of vascular grafts of 1 cm in length to abdominal aortae of rat, endothelial cells can be formed easily by stretching anastomosis. We evaluated the endothelialization in the center part of vascular grafts by implanting vascular grafts using transgenic (TG) silk fibroin (SF) of 3 cm in length. Vascular grafts were prepared 1.5 mm in diameter and 1 and 3 cm in length using wild type (WT) SF and TG SF by braiding structure, respectively. The grafts were removed after 2 weeks or 3 months and evaluated pathologically. Endothelialization was not confirmed totally after 3 months of implantation. However, endothelialization in the center part of grafts was significantly higher in TG SF than in WT SF. No significant difference was found regarding tissue infiltration and internal diameter. The TG SF revealed migration of the endothelial cells into the center part of the vessels at the early stage. Also, tissue infiltration and remodeling is expected using SF. The 3 cm length vascular grafts can be evaluated as a new experimental system.

Silk fibroin-Pellethane® cardiovascular patches: Effect of silk fibroin concentration on vascular remodeling in rat model.

Life-threatening cardiovascular anomalies require surgery for structural repair with cardiovascular patches. The biomaterial patch, derived from Bombyx mori silk fibroin (SF), is used as an alternative material due to its excellent tissue affinity and biocompatibility. However, SF lacks the elastomeric characteristics required for a cardiovascular patch. In order to overcome this shortcoming, we combined the thermoplastic polyurethane, Pellethane® (PU) with SF to develop an elastic biocompatible patch. Therefore, the purpose of this study was to investigate the feasibility of the blended SF/PU patch in a vascular model. Additionally, we focused on the effects of different SF concentrations in the SF/PU patch on its biological and physical properties. Three patches of different compositions (SF, SF7PU3 and SF4PU6) were created using an electrospinning method. Each patch type (n = 18) was implanted into rat abdominal aorta and histopathology was assessed at 1, 3, and 6 months post-implantation. The results showed that with increasing SF content the tensile strength and elasticity decreased. Histological evaluation revealed that inflammation gradually decreased in the SF7PU3 and SF patches throughout the study period. At 6 months post-implantation, the SF7PU3 patch demonstrated progressive remodeling, including significantly higher tissue infiltration, elastogenesis and endothelialization compared with SF4PU6. In conclusion, an increase of SF concentration in the SF/PU patch had effects on vascular remodeling and physical properties. Moreover, our blended patch might be an attractive alternative material that could induce the growth of a neo-artery composed of tissue present in native artery.