Novel Zinc Finger Transcription Factor ZFP580 Facilitates All-Trans Retinoic Acid -Induced Vascular Smooth Muscle Cells Differentiation by Rarα-Mediated PI3K/Akt and ERK Signaling.

BACKGROUND/AIMS: Phenotypic switching of vascular smooth muscle cells (VSMC) plays a vital role in the development of vascular diseases. All-trans retinoic acid (ATRA) is known to regulate VSMC phenotypes. However, the underlying mechanisms remain completely unknown. Here, we have investigated the probable roles and underlying mechanisms of the novel C2H2 zinc finger transcription factor ZFP580 on ATRA-induced VSMC differentiation. METHODS: VSMCs were isolated, cultured, and identified. VSMCs were infected with an adenovirus encoding ZFP580 or Ad-siRNA to silence ZFP580. The expression levels of ZFP580, SMα-actin, SM22α, SMemb, RARα, RARß, and RARγ were assayed by Q-PCR and western blot. A rat carotid artery injury model and morphometric analysis of intimal thickening were also used in this study. RESULTS: ATRA caused a significant reduction of VSMC proliferation and migration in a doseand time-dependent manner. Moreover, it promoted VSMC differentiation by enhancing expression of differentiation markers and reducing expression of dedifferentiation markers. This ATRA activity was accompanied by up-regulation of ZFP580, with concomitant increases in RARα expression. In contrast, silencing of the RARα gene or inhibiting RARα with its antagonist Ro41-5253 abrogated the ATRA-induced ZFP580 expression. Furthermore, ATRA binding to RARα induced ZFP580 expression via the PI3K/Akt and ERK pathways. Adenovirusmediated overexpression of ZFP580 promoted VSMC differentiation by enhancing expression of SM22α and SMα-actin and reducing expression of SMemb. In contrast, silencing ZFP580 dramatically reduced the expression of differentiation markers and increased expression of dedifferentiation markers. The classic rat carotid artery balloon injury model demonstrated that ZFP580 inhibited proliferation and intimal hyperplasia in vivo. CONCLUSION: The novel zinc finger transcription factor ZFP580 facilitates ATRA-induced VSMC differentiation by the RARα-mediated PI3K/Akt and ERK signaling pathways. This might represent a novel mechanism of regulation of ZFP580 by ATRA and RARα, which is critical for understanding the biological functions of retinoids during VSMC phenotypic modulation.

Salpingopharyngeal fistula as a treatment for guttural pouch mycosis in seven horses.

BACKGROUND: Guttural pouch mycosis (GPM) is a cause of nasal discharge, dysphagia and fatal haemorrhage in the horse. OBJECTIVES: To report the complications and success of salpingopharyngeal fistulation in horses with GPM. We hypothesised that creating a direct static opening into the guttural pouch from the pharynx would cause a regression of fungal plaques due to a change in the guttural pouch environment and that this treatment would result in resolution of infection prior to secondary complications. STUDY DESIGN: Retrospective case series. METHODS: The medical records of all horses diagnosed with GPM that were presented to New Bolton Center between the years 2006 and 2017 were examined retrospectively. Seven cases of guttural pouch mycosis treated with salpingopharyngostomy without other surgical intervention were included. Information collected included signalment, presenting complaint, which pouch was affected, size and location of the plaques, laryngeal and pharyngeal function, concurrent medical therapy, location of the fistula, surgical time, time to resolution of clinical signs, time to full resolution of the mycotic plaque, and patency of the salpingopharyngostomy site. RESULTS: Nasal discharge resolved in 10-30 days post-operatively in all cases where nasal discharge was present. The mycotic plaques showed complete resolution at time points ranging from 1 to 6 months post-operatively. No case developed epistaxis or neurological deficits post-operatively that were not present at presentation. MAIN LIMITATIONS: There were differing adjunctive treatments between cases. This technique is not appropriate for horses that have had epistaxis or are currently bleeding. CONCLUSIONS: Salpingopharyngostomy can minimise cost of treatment, be performed on an outpatient basis and provide better exposure of the infected area with few complications. This case series documents seven cases treated with this method that resolved the infection without any further complications of the mycosis.

Retropharyngeal abscess with carotid involvement presenting as fever of undetermined origin in a dog.

A young dog was presented with lethargy and pyrexia of 2 days duration, not responding to empirical treatment. Thorough diagnostic investigation failed to determine the cause. A retropharyngeal abscess became apparent when it eroded into the carotid artery 2 days later. This case highlights the challenging nature of fever of undetermined origin (FUO) and the value of close monitoring for diagnosis and prompt intervention.

Abcès rétro-pharyngien avec une atteinte de la carotide et une fièvre d'origine indéterminée chez un chien. Un jeune chien a été présenté pour un abattement et une pyrexie d'une durée de 2 jours et il ne répondait pas au traitement empirique. Une enquête diagnostique complète n'a pas réussi à déterminer la cause. Un abcès rétro-pharyngien est devenu apparent lorsqu'il s'est érodé dans la carotide 2 jours plus tard. Ce cas souligne la nature difficile d'une fièvre d'origine indéterminée et la valeur d'une étroite surveillance pour le diagnostic et une intervention rapide.(Traduit par Isabelle Vallières).

Trans-arterial coil embolization of the internal carotid artery in standing horses.

OBJECTIVES: To develop transarterial coil embolization (TACE) for occlusion of the internal carotid artery (ICA), in normal standing horses, and to evaluate it use for prevention of hemorrhage in horses with guttural pouch mycosis (GPM). STUDY DESIGN: Prospective study. ANIMALS: Normal horses (n = 8) and 5 with GPM. METHODS: Horses had TACE of the ICA in standing position under fluoroscopic guidance. Four normal horses were euthanatized 2 weeks after TACE for morphologic assessment and 4 were followed for 6 months. The 5 clinically affected horses were evaluated for long-term (10-12 months) success rate and complications. RESULTS: No complications related to the TACE were noted. Up to 30 mL warmed meglumine ioxithalamate was injected and well tolerated. Standing angiography confirmed complete occlusion of all vessels, and coils were positioned as intended; the procedure did not alter local hemodynamics. At 2 weeks, maturing to mature continuous thrombi was seen at the site of the coils. Two clinically affected horses died at day 3 and 12 after surgery from other problems. In the 3 surviving horses, mycotic lesions completely resolved without additional treatment. CONCLUSIONS: TACE under fluoroscopic guidance in standing horses provided a safe, minimally invasive, and effective method for ICA occlusion and should be recommended for individuals at risk of general anesthesia. Residual neurologic deficits are a common sequela, but they do not reflect a treatment failure.

Efecto vasodilatador in vitro de la cafeína en aorta de conejos ateroscleróticos / In-vitro vasodilator effect of caffeine in atherosclerotic aorta rabbits

Introducción. Se desconocen los efectos directos de la cafeína en arterias con disfunción endotelial y aterosclerosis. Objetivo. Evaluar los efectos vasoactivos in vitro de la cafeína en anillos de aorta de conejos ateroscleróticos. Metodología. La aterosclerosis fue inducida en conejos (n = 10) alimentados con dieta aterogénica (1% colesterol, durante 16 semanas) (grupo 1). El grupo control (n = 10) recibió una dieta estándar libre de colesterol (grupo 2). Al final de las 16 semanas, se sacrificó a los animales y se determinó el colesterol sérico. Los estudios de reactividad vascular y análisis morfométrico se llevaron a cabo en anillos de aorta torácica. Se estudió la reactividad vascular en respuesta a la acetilcolina, la nitroglicerina y la cafeína a 3 dosis correspondientes al contenido de esta sustancia de 1, 2 y 3 triples cafés expresso. Resultados. La relajación máxima a la acetilcolina en arterias de conejos sanos (22,5 ± 16,8%) fue mayor que en arterias de conejos ateroscleróticos (3,6 ± 3,7%; p = 0,006). Aunque el efecto vasodilatador de la cafeína fue dependiente de la concentración (p < 0,001), no se encontraron diferencias entre las arterias provenientes de conejos sanos (75,73 ± 11,20%) y las de conejos ateroscleróticos (68,19 ± 15,07%; p = 0,238). La nitroglicerina produjo una relajación menor que la cafeína, tanto en arterias de conejos sanos (32,78 ± 12,30%), como en las de conejos enfermos (73,48 ± 18,93%; p < 0,001). El EC50 (concentración del agente vasodilatador que causa un 50% de relajación) fue similar para los 2 vasodilatadores (p = 0,178). Las lesiones aórticas en el grupo 1 consistieron en placas tempranas. El recubrimiento endotelial (CD31) fue del 92,2 ± 5,6% y el 92 ± 4,8%, respectivamente (p = 0,927). Conclusiones. La cafeína ejerce un potente efecto vasodilatador arterial in vitro, independiente de la presencia o la ausencia de aterosclerosis (AU)

Introduction. The effects of caffeine on arteries with endothelial dysfunction and atherosclerosis are unknown. Objective. To evaluate the in-vitro vasoactive effects of caffeine on aortic rings from atherosclerotic rabbits. Methodology. Atherosclerosis was induced in rabbits (n = 10) fed an atherogenic diet (1% cholesterol) (group 1). The control group (n = 10) received a cholesterol-free diet (group 2). At 16 weeks we evaluated serum cholesterol, and all the animals were sacrificed. Thoracic aorta rings were obtained for vasoreactivity studies and morphometric analyses. Agonists were nitroglycerine, acetylcholine, and caffeine at 3 doses corresponding to one, 2 and 3 triple espressos. Results. Arterial relaxation with acetylcholine in arteries from healthy rabbits (22.5 ± 16.8%) was greater than in arteries from atherosclerotic rabbits (3.6 ± 3.7%; p = 0.006). Although the vasodilator effect of caffeine was dependent on the concentration (p < 0.001), no differences were found between arteries from healthy rabbits (75.73 ± 11.20%) and those from diseased rabbits (68.19 ± 15.07%; p = 0.238). Nitroglycerine generated less relaxation than caffeine, both in arteries from healthy rabbits (32.78 ± 12.30%) and from diseased rabbits (73.48 ± 18.93%; p < 0.001). The EC50 (half maximal effective concentration) was similar for both vasodilators (p = 0.178). The aortic lesions in group 1 consisted of early plaques. The endothelial covering (CD31) was 92.2 ± 5.6% and 92 ± 4.8% respectively (p = 0.927). Conclusions. Caffeine exerts a potent arterial vasodilator effect in-vitro regardless of the presence or absence of aterosclerosis (AU)

Tenascin-C-coated platinum coils for acceleration of organization of cavities and reduction of lumen size in a rat aneurysm model.

OBJECT: Detachable platinum coils are widely used in the endovascular treatment of intracranial aneurysms. The use of coil placement produces a higher incidence of aneurysm recurrence compared with surgical clipping. To reduce the incidence of recurrence by promoting clot organization, the authors designed a platinum coil coated with tenascin-C (TNC), an extracellular matrix glycoprotein, and then histologically examined tissue responses. METHODS: Platinum coils were prepared by successive coatings with cationic polyethyleneimine and anionic heparin and then TNC or basic fibroblast growth factor (bFGF) was immobilized by affinity binding to the heparin. Six unmodified, six heparin-coated, six bFGF-coated, or eight TNC-coated platinum coils were inserted into ligated common carotid arteries (CCAs) of adult male rats, and CCA segments were harvested after 14 or 28 days. The percentages of organized areas occupying the luminal cavity in unmodified, heparin-coated, bFGF-coated, and TNC-coated groups were 4.8 +/- 4.6, 1.6 +/- 1.1, 17.9 +/- 10.7, and 93.4 +/- 6.9%, respectively. In addition, the mean lumen size in the TNC-coated group (0.35 +/- 0.23 mm2) was reduced to less than half that of the unmodified group (0.72 +/- 0.21 mm2). Immunohistochemical analysis revealed that alpha-smooth muscle actin-positive cells were a major cellular component of the organized tissue within the TNC-coated coils but not in the bFGF group. Collagen fibrils in the organized areas were also much thicker and denser with TNC-coated coils than with bFGF-coated coils. CONCLUSIONS: Placement of TNC-coated coils can remarkably accelerate organization of luminal cavities and reduce their volume, providing improved efficacy of these coils for endovascular embolization.

Treatment of mechanically-induced vasospasm of the carotid artery in a primate using intra-arterial verapamil: a technical case report.

BACKGROUND: Despite improvements in the safety and efficacy of endovascular procedures, considerable morbidity may still be attributed to vasospasm. Vasospasm has proven amenable to pharmacological intervention such as nitrates, intravenous calcium channel blockers (CCBs), and intra-arterial papaverine, particularly in small vessels. However, few studies have focused on medium to large vessel spasm. Here we report the use of an intra-arterial CCB, verapamil, to treat flow-limiting mechanically-induced spasm of the common carotid artery (CCA) in a primate. We believe this to be the first such report of its kind. CASE PRESENTATION: As part of a study assessing the placement feasibility and safety of a catheter capable of delivering intra-arterial cerebroprotective therapy, a female 16 kg baboon prophylaxed with intravenous nitroglycerin underwent transfemoral CCA catheterization with a metallic 6-Fr catheter without signs of acute spasm. The protocol dictated that the catheter remain in the CCA for 12 hours. Upon completion of the protocol, arteriography revealed a marked decrease in CCA size (mean cross-sectional area reduction = 31.6 +/- 1.9%) localized along the catheter length. Intra-arterial verapamil (2 mg/2cc) was injected and arteriography was performed 10 minutes later. Image analysis at 6 points along the CCA revealed a 21.0 +/- 1.7% mean increase in vessel diameter along the length of the catheter corresponding to a 46.7 +/- 4.0% mean increase in cross-sectional area. Mean systemic blood pressure did not deviate more than 10 mm Hg during the procedure. CONCLUSIONS: Intraluminal CCBs like verapamil may constitute an effective endovascular treatment for mechanically-induced vasospasm in medium to large-sized vessels such as the CCA.

Fluorangiographic alterations in a dog with suspected iatrogenic partial carotid artery occlusion.

During routine fluorescein angiography, a healthy female dog demonstrated abnormal delayed filling of retinal and choroidal vessels. Fluorangiographic times were increased four- to fivefold with respect to reference values. Fluorescein angiography, using the same protocol, was repeated two weeks later and a fluorangiogram within reference values was obtained. The abnormal results in the first procedure were suspected to have resulted from partial occlusion of the carotid artery secondary to the restraining method used.

Dissecting aneurysm of the carotid artery as a cause of respiratory distress in adult cattle.

A two-and-a-half-year-old Friesian cow and a five-year-old Charolais cow developed severe respiratory distress and palpable swellings to the left of the larynx as a result of a dissecting aneurysm of the common carotid artery. Neither cow responded to medical treatment. The underlying pathogenesis of the condition was uncertain, but direct trauma to the carotid artery was a possible contributory factor. Aneurysms of the common carotid artery should be considered when swelling occurs in the region of the larynx or when respiratory distress is due to laryngeal compression.

Histologic features of the carotid artery trifurcation in thoroughbreds.

The common, external, and internal carotid and occipital arteries were examined histologically at the trifurcation of the common carotid arteries in 13 Thoroughbred foals (0 to 30 days old) and 64 Thoroughbred adults (2 to 4 years old). Calcification in the media of the common carotid and external carotid arteries was observed in 3 of the 13 foals and in 30 of the 64 adult horses. Calcification resembled that seen in Mönckeberg's arteriosclerosis in human beings, the cause of which is unknown.

Aneurysms in a large colony of squirrel monkeys (Saimiri sciureus).

This communication reports the findings of a retrospective study of extracranial aneurysms found at necropsy in a large colony of squirrel monkeys (Saimiri sciureus). Eleven (1.5%) of 730 cases had dissecting, saccular, or fusiform aneurysms of the carotid arteries or aorta. Saccular and fusiform aneurysms were found only in animals that had been fed atherogenic diets, whereas dissecting aneurysms occurred in both normo- and hypercholesterolemic monkeys. Neither the type or location of aneurysms, however, could be predicted by the length of time an animal consumed an atherogenic diet, nor by the total mean serum cholesterol concentration. The anatomical characteristics, location, and incidence of aneurysms found in squirrel monkeys resembled closely those observed in human autopsy cases.

Occlusion of internal carotid artery in the horse by means of a balloon-tipped catheter: evaluation of a method designed to prevent epistaxis caused by guttural pouch mycosis.

A procedure developed to occlude the internal carotid artery of horses with guttural pouch mycosis was modified was modified and evaluated in nine clinically normal horses. The left internal carotid artery was ligated at its origin and occluded distally with an intraluminal balloon catheter. In four horses, the balloon-tipped catheters were left in place until these horses were euthanatized at 4 to 244 days after surgery. In the remaining horses, the catheters were removed after 10 to 14 days by a cutdown procedure, and these horses were euthanatized 1 to 60 days after catheter removal. In both groups, gross and histologic examinations revealed that occlusive thrombosis had developed in sections of artery distal to the balloon. Postoperative complications were not encountered.

Occlusion of internal carotid artery in the horse by means of a balloon-tipped catheter: clinical use of a method to prevent epistaxis caused by guttural pouch mycosis.

An intravascular procedure was used to occlude the internal carotid artery of two horses with epistaxis caused by guttural pouch mycosis. In each horse, the affected internal carotid artery was ligated close to its origin. A balloon-tipped catheter was introduced into the artery distal to the ligature, and its tip was advanced beyond the site of infection. The balloon was then inflated so that the infected segment of artery was isolated from the cerebral vascular system. The catheters were removed at 14 and 51 days, and both horses were returned to training and racing. Neither horse had further episodes of epistaxis.