RESUMEN
Anesthesia strongly influences laboratory animals, and it can also greatly affect the experimental data. Rats rank only second to mice in the number used in research fields, such as organ transplantation, regenerative medicine and imaging. Therefore, appropriate and effective anesthesia, including the protocol of the endotracheal intubation and inhalation anesthesia, is crucial. Hence, we evaluated these methods in this study. Twelve Wistar rats were intraperitoneally injected with M/M/B: 0.3/4/5, comprising of medetomidine, midazolam and butorphanol at a dose of 0.3 mg/kg + 4.0 mg/kg + 5.0 mg/kg body weight/rat, respectively. An endotracheal tube was then intubated into the trachea. After intubation, the rats were connected to the inhalation anesthesia circuit using isoflurane, and vital signs were measured until 30 min after connection. All intubations were successfully finished within 1 min, and the values of the vital signs were normal and stable. In addition, histopathological observation of the trachea and lungs showed no trauma. These results suggest that this visible endotracheal intubation method is simple, reliable, safe and favorable with regard to the rats' welfare.
Asunto(s)
Anestesia por Inhalación/instrumentación , Anestésicos por Inhalación/administración & dosificación , Intubación Intratraqueal/métodos , Isoflurano/administración & dosificación , Laringoscopios , Anestesia por Inhalación/métodos , Anestésicos por Inhalación/farmacología , Animales , Femenino , Frecuencia Cardíaca , Intubación Intratraqueal/efectos adversos , Isoflurano/farmacología , Masculino , Oxígeno/sangre , Ratas , Ratas Wistar , Frecuencia RespiratoriaRESUMEN
Appropriate and effective anesthesia is critical, because it has a strong influence on laboratory animals, and its affect greatly impacts the experimental data. Inhalational anesthesia by endotracheal intubation is currently prevailing in general anesthesia and is prefered over injection anesthesia, especially for large laboratory animals, because it is a safe and easy control agent. However, it is not common for small laboratory animals, because of the high degree of technical skills required. We assessed the capability of use for mice of the endotracheal intubation by using the endoscope system "TESALA AE-C1" and inhalational anesthesia using a ventilator. Endotracheal intubation was successfully performed on all 10 C57BL/6 mice injected with M/M/B: 0.3/4/5 comprised of medetomidine, midazoram and butorphanol, at a dose of 0.3 mg/kg + 4.0 mg/kg + 5.0 mg/kg body weight/mouse, respectively. After the intubated mice were connected with the inhalational anesthesia circuit and the ventilator, vital signs were measured until 15 min after the connection. The data with M/M/B: 0.3/4/5 showed stable and normal values, which indicated that this new endotracheal intubation method was simple, reliable and safe, which mean that this anesthesia is favorable in regard to the animal's welfare.
Asunto(s)
Anestesia por Inhalación/instrumentación , Anestesia por Inhalación/métodos , Intubación Intratraqueal/métodos , Animales , Endoscopía/métodos , Femenino , Masculino , RatonesRESUMEN
Immunodeficient animals are in demand in current biomedical research, and they contribute to medical progress. In Pneumocystis infections, a specific histological diagnostic tool may be immunochemistry (IC). However, it was recently reported that the antibody (3F6) was not suitable for detecting Pneumocystis in rats. We purchased another antibody [PNC007] from a commercial source for IC. We could detect positive signals at identical locations with IC and Toluidine blue O in lungs of infected rats. These results corresponded to the results obtained with PCR. We should study the relationship between unexpected positive signals seen in IC and trophic forms in lungs of infected rats. We could clinically diagnose pneumonia caused by Pneumocystis carinii with the diagnostic method we developed.
Asunto(s)
Huésped Inmunocomprometido , Inmunohistoquímica/métodos , Pneumocystis carinii/aislamiento & purificación , Neumonía por Pneumocystis/diagnóstico , Animales , Anticuerpos Antifúngicos/inmunología , Femenino , Pulmón/inmunología , Pulmón/microbiología , Pulmón/patología , Pneumocystis carinii/inmunología , Neumonía por Pneumocystis/inmunología , Neumonía por Pneumocystis/microbiología , Ratas , Ratas Endogámicas F344 , Ratas WistarRESUMEN
The Toyobo-National Cardiovascular Center pneumatic ventricular assist device (Toyobo-NCVC VAD) is widely used in Japan; however, the current pneumatic drivers have some drawbacks, including their large size, heavy weight, and high power consumption. These issues cause difficulty with mobility and contribute to an unsatisfactory quality of life for patients. Because it is urgently necessary to improve patients' safety and quality of life, we have developed a compact, low-noise, portable VAD driver by utilizing an electrohydraulic actuator consisting of a brushless DC motor and a regenerative pump. This unit can be actuated for as long as 2 h with two rechargeable lightweight batteries as well as with external AC power. It is compact in size (33 x 25 x 43 cm) and light in weight (13 kg), and the unit is carried on a mobile wheeled cart. In vitro testing with a Toyobo-NCVC VAD demonstrated a sufficient pumping capacity of up to 8 l/min. We conclude that this newly-developed compact portable driver can provide a better quality of life and improved safety for patients using protracted pneumatic VAD support.
Asunto(s)
Suministros de Energía Eléctrica , Corazón Auxiliar , Electrónica Médica , Humanos , Miniaturización , Diseño de PrótesisRESUMEN
We developed a new coating material (Toyobo-National Cardiovascular Center coating) for medical devices that delivers high antithrombogenicity and long-term durability. We applied this coating to an extracorporeal membrane oxygenation (ECMO) system, including the circuit tube, cannulae, a seal-less centrifugal pump, and a diffusion membrane oxygenator, to realize prolonged cardiopulmonary support with trivial anticoagulant infusion. The oxygenator consisted of a hollow-fiber membrane made of polymethylpentene, which allows the transfer of gas by diffusion through the membrane. The centrifugal pump was free of seals and had a pivot bearing. We performed a venoarterial bypass in a goat using this ECMO system, and the system was driven for 151 days with trivial anticoagulant infusion. Plasma leakage from the oxygenator did not occur and sufficient gas-exchange performance was well maintained. In the oxygenator, thrombus formation was present around the top and the distributor of the inlet portion and was very slight in the outlet portion. In the centrifugal blood pump, there was some wear in the female pivot region and quite small amounts of thrombus formation on the edge of the shroud; the pivot wear seemed to be the cause of the hemolysis observed after 20 weeks of perfusion and which resulted in the termination of the perfusion. However, no significant amounts of thrombus were observed in other parts of the system. This ECMO system showed potential for long-term cardiopulmonary support with minimal use of systemic anticoagulants.
Asunto(s)
Anticoagulantes/administración & dosificación , Materiales Biocompatibles Revestidos , Oxigenación por Membrana Extracorpórea/instrumentación , Heparina/administración & dosificación , Perfusión/instrumentación , Animales , Centrifugación , Modelos Animales de Enfermedad , Diseño de Equipo , Cabras , Factores de TiempoRESUMEN
Aortic wall thinness was one of the most characteristic changes observed in experimental animals under prolonged continuous flow left heart bypass. The goal of this study was to determine the roles of smooth muscle cells in the vascular remodeling process in cases demonstrating aortic wall thinness under prolonged continuous flow left heart bypass. The aortic samples from three goats in which continuous flow left heart bypass was performed were subjected to histological and immunohistochemical analyses. After 4 weeks of observation, the pulse pressure in the goats under the continuous flow left heart bypass was clearly lower than that in the normal healthy goats. The aortic walls of these goats became thinner, an effect caused by the dilation of their internal diameter. These aortic smooth muscle cells maintained contractile formation due to the fact that they contained abundant alpha-smooth muscle actin (SMA) and smooth muscle myosin heavy chain (SMMS). These cells also synthesized redundant matrix metalloproteinase-2 and -9, and the ratio of the SMMS-positive to the SMA-positive area was significantly lower (0.76) than that observed in the control goat (1.00; P < 0.05). The smooth muscle cells demonstrated synthetic-dedifferentiated formation, which is one of the phenotypes of smooth muscle cell function. In conclusion, aortic wall thinness under prolonged continuous flow left heart bypass is caused by over-synthesis of matrix metalloproteinase in smooth muscle cells, and this refers the vascular remodeling process of the extracellular matrix.
Asunto(s)
Puente Cardíaco Izquierdo , Metaloproteinasas de la Matriz/biosíntesis , Músculo Liso Vascular/citología , Animales , Aorta/citología , Aorta/patología , Cabras , InmunohistoquímicaRESUMEN
The integrated heart lung assist device (IHLAD) has been developed to overcome the problems of currently available extracorporeal membrane oxygenation devices. The integrated structure of a centrifugal blood pump and cylindrical bundle of polyolefin hollow-fibers has allowed a remarkably compact size for the device. This study deals with the design change of the IHLAD that added to the vaned diffuser between the impeller of the centrifugal pump and the hollow-fiber bundle with a view to enhancing the gas-transfer performance. Ex vivo gas-transfer performance tests were carried out, as well as hydrodynamic characteristics and hemolysis test using fresh goat blood. The oxygen transfer rate was generally improved, and the carbon dioxide removal rate was slightly improved. Intolerable amount of hemolysis (index of hemolysis= 0.177) was caused by the IHLAD, which must be resolved by improving the design in the future.
Asunto(s)
Corazón Auxiliar , Máquina Corazón-Pulmón , Animales , Dióxido de Carbono/sangre , Diseño de Equipo , Cabras , Ensayo de Materiales , Membranas Artificiales , Oxígeno , Oxigenadores de MembranaRESUMEN
Electrohydraulic total artificial heart (EHTAH) and electrohydraulic ventricular assist device (EHVAD) systems have been developed in our institute. The EHTAH system comprises a pumping unit consisting of blood pumps and an actuator, as well as an electronic unit consisting of an internal controller, internal and external batteries, and transcutaneous energy transfer (TET) and optical telemetry (TOT) subunits. The actuator, placed outside the pericardial space, reciprocates and delivers hydraulic silicone oil to the alternate blood pumps through a pair of flexible oil conduits. The pumping unit with an external controller was implanted in 10 calves as small as 55 kg. Two animals survived for more than 12 weeks in a good general condition. The assumed cardiac output ranged between 6 and 10 L/min, the power consumption was 12-18 W, and the energy efficiency was estimated to be 9-11%. Initial implantation of subtotal system including electronic units was further conducted in another calf weighing 73 kg. It survived for 3 days with a completely tether free system. The EHVAD system is developed by using the left blood pump and the actuator of the EHTAH, which were packaged in a compact metal casing with a compliance chamber. In vitro testing demonstrated maximum output more than 9 L/min and more than 13% maximum efficiency. The initial animal testing lasted for 25 days. These results indicate that our EHTAH and EHVAD have the potential to be totally implantable systems.
Asunto(s)
Corazón Artificial , Animales , Gasto Cardíaco/fisiología , Bovinos , Diseño de Equipo , Corazón Artificial/efectos adversos , Tromboembolia/etiologíaRESUMEN
In the mechanism of damage to red blood cells (RBCs) caused by a centrifugal pump, the prolonged effects to the RBC membrane caused by exposure to shear stress remain unclear. We focused on the band 3 protein (B3), one of the major proteins in the membrane skeleton, and investigated the ultrastructural alterations of the RBC membrane with loaded shear stress. Using flow cytometry, the relative amount of B3 was examined in relation to RBC deformability. The results, with continuous exposure to low shear stress, showed cell downsizing, an increase in B3 density, and a decrease in the deformability of the RBC membrane. Exposure to high shear stress does not appear to exert any influence on the membrane skeleton of the RBC. Therefore, in addition to conventional processes including the instantaneous destruction of a cell due to intense shear stresses, the results of the present study indicate the presence of another process based on changes in membrane proteins leading to cell fragmentation. Under low shear stress, the RBC membrane skeleton shows delayed destruction, which is exhibited as a disorder of B3 distribution, and the related membrane dysfunction includes decreases in RBC deformability and stability.
Asunto(s)
Deformación Eritrocítica/fisiología , Membrana Eritrocítica/patología , Membrana Eritrocítica/fisiología , Animales , Viscosidad Sanguínea , Citoesqueleto/fisiología , Membrana Eritrocítica/ultraestructura , Citometría de Flujo , Cabras , Hemólisis , Membrana Dobles de Lípidos , Estrés MecánicoRESUMEN
We investigated morphological changes of the arterial systems in the kidneys under prolonged continuous flow left heart bypass. Twelve goats were subjected to 2 weeks of pulsatile left heart bypass followed by 4 weeks of continuous flow left heart bypass (group CF). After autopsy, the kidneys underwent pathological evaluation. Six normal healthy goats were used as controls. The media of the afferent arterioles of group CF were frequently thickened by an increase in the number of the mature smooth muscle cells (SMCs). The juxtaglomerular areas (JGA) were expanded because of an increase in the number and size of SMCs and/or SMC-like cells. Furthermore, the percentage of anti-proliferating cell nuclear antigen antibody-positive cells in the JGA of group CF (9.9 +/- 1.9%) was significantly higher (p = 0.025) than that of the control group (4.6 +/- 3.4%), indicating active proliferation in group CF. We concluded that prolonged continuous flow left heart bypass causes proliferation of SMCs and/or SMC-like cells in the afferent arterioles and their perivascular tissue.
Asunto(s)
Enfermedades Cardiovasculares/fisiopatología , Enfermedades Cardiovasculares/terapia , Puente Cardíaco Izquierdo , Arteria Renal/patología , Animales , Modelos Animales de Enfermedad , Cabras , Hemodinámica/fisiología , Aparato Yuxtaglomerular/patología , Aparato Yuxtaglomerular/fisiopatología , Flujo Pulsátil/fisiología , Arteria Renal/fisiopatología , Circulación Renal/fisiología , Factores de Tiempo , Túnica Media/patología , Túnica Media/fisiopatologíaRESUMEN
Aortic pressure declines during cardiopulmonary bypass (CPB), particularly at normothermia. It has been reported that administering vasoconstrictors during normothermic CPB (NCPB) to restore perfusion pressure might induce hypoperfusion of splanchnic organs. We have reported that prostaglandin (PG), metabolized in the lung but increased during CPB, might have played a substantial role in hypotension, and that a PG synthesis inhibitor (PGSI) could improve hypotension during CPB. This study was designed to examine whether regional perfusion of splanchnic organs was reduced when PGSI restored systemic perfusion pressure during NCPB. NCPB was performed in eight adult goats for 60 minutes (body weight 57.0 +/- 5.9 kg). PGSI was administered in group P (n = 4), while norepinephrine was administered in group C (n = 4), to keep aortic pressure in the range of 50 to 80 mm Hg. The total systemic flow was maintained at approximately 70 ml/kg/min. Tissue blood flow was measured by means of the colored microsphere method before and 30 and 60 minutes after the start of CPB. In group P, gut blood flows after the start of CPB were higher than those before CPB, significantly in the stomach and jejunum at 30 minutes (p < 0.05), whereas gut blood flows in group C were decreased or not changed. In conclusion, PGSI prevents hypotension without impairing gut perfusion during NCPB.
Asunto(s)
Puente Cardiopulmonar/métodos , Inhibidores de la Ciclooxigenasa/farmacología , Flurbiprofeno/farmacología , Hipotensión/prevención & control , Prostaglandinas/biosíntesis , Circulación Esplácnica , Animales , Presión Sanguínea , Temperatura Corporal , Cabras , Yeyuno/irrigación sanguínea , Pulmón/metabolismo , Microesferas , Norepinefrina/farmacología , Oxígeno/metabolismo , Estómago/irrigación sanguínea , Vasoconstrictores/farmacologíaRESUMEN
We developed an extracorporeal membrane oxygenation (ECMO) system with high antithrombogenicity and durability characteristics for prolonged continuous cardiopulmonary support. The oxygenator consists of a special hollow-fiber-type polyolefin gas-exchange membrane, which has an ultrathin dense layer in contact with the blood, in order to prevent plasma leakage during protracted use (Platinum Cube NCVC). The centrifugal pump (RotaFlow) is free of seals. The entire blood-contacting surface of the system is coated with a newly developed heparin material (Toyobo-NCVC coating). We performed a venoarterial bypass in a goat, and the ECMO system was driven for 34 days without systemic anticoagulants. Plasma leakage from the oxygenator did not occur, and sufficient gas exchange performance was maintained. Thrombus formation was hardly observed in the ECMO system except in the casing margins of the oxygenator. This ECMO system showed potential for long-term cardiopulmonary support with minimal or no use of systemic anticoagulants.