Sympathetic control of substrates and thermogenesis during central cooling.
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Animals↗
Biomedical subjects
K Muramoto
Publications and source records attributed to K Muramoto.
Lectins in the hemolymph of a starfish, Asterina pectinifera: purification and characterization.
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Agglutination Tests↗
Photoaffinity labeling of an acorn barnacle lectin with a photoactivatable fluorescent reagent derivative of D-galactosamine.
A photoactivatable D-galactosamine derivative was prepared by reaction of the amino group of D-galactosamine with 1-azide-5-naphthalene sulfonyl chloride (ANS-Cl). The derivative (GalN-ANS) inhibited the agglutination activity of an acorn barnacle lectin against rabbit erythrocytes to the same extent as D-galactosamine. We used GalN-ANS for photoaffinity labeling of the lectin. The photolabeled lectin was digested with pronase and the digest was separated by reversed-phase high-performance liquid chromatography by monitoring fluorescence and uv absorption to isolate the peptide labeled with GalN-ANS. Amino acid analyses of the labeled peptides revealed that GalN-ANS preferentially covalently labeled two regions in the carbohydrate recognition domain of the lectin. One of them was the highly conserved amino-acid sequence region throughout all calcium-dependent animal lectins.
Affinity Labels↗
Purification and characterization of an agglutinin of the soft coral Sinularia species.
A D-galactose-specific agglutinin, named sinularian, has been isolated from the soft coral Sinularia sp. by affinity chromatography on acid-treated Sepharose 4B and by gel filtration on HPLC. Sinularian was a glycoprotein containing 11% sugar. It gave a single band corresponding to 78 kDa in SDS-PAGE, irrespective of a treatment with 2-mercaptoethanol. Sinularian agglutinated rabbit erythrocytes and murine leukemia cells but not sheep or human ABO erythrocytes. Its hemagglutinating activity was Ca(++)-independent. Sinularian promoted binding of macrophages to tumor cells.
Agglutinins↗
Basal ganglia neural activity during operant feeding behavior in the monkey: relation to sensory integration and motor execution.
The activity of single neurons in the caudate nucleus (CD), globus pallidus (GP), substantia nigra pars reticulata (SNr), and ventral tegmental area (VTA) was recorded during an operant feeding task in the monkey. The task had three phases: recognition of the food or nonfood stimulus (1st phase), bar pressing to obtain access to the stimulus (2nd phase), and ingestion (3rd phase). Data were collected from 351 neurons in CD, 344 in GP, 261 in SNr, and 275 in VTA. Neurons in the dorsolateral part of the CD, GP, and SNr responded primarily to motor events of feeding, i.e., extension/flexion of the arm, bar pressing, chewing, grasping or gazing. Neurons in the ventromedial part of the CD and rostroventral part of the GP exhibited differential responses to the presentation of food and nonfood during the recognition and bar pressing phases of the task. Neurons in the VTA increased their firing early in the bar pressing phase and then decreased their firing during ingestion. The data suggest that the dorsolateral part of the basal ganglia is involved mainly in motor function, while the ventromedial part may reflect the connection between motivation and motor output.
Animals↗
Galectin containing cells in the skin and mucosal tissues in Japanese conger eel, Conger myriaster: an immunohistochemical study.
Congerin is a beta-galactoside binding lectin (galectin) purified from the skin mucus of the Japanese conger, Conger myriaster. To clarify its tissue distribution and productive cells, several tissue samples including skin, buccal cavity wall, tang, pharynx, gills, esophagus, stomach, intestine, liver, kidney, spleen and ovary of conger were stained immunohistochemically using polyclonal rabbit anti-congerin serum. In the epidermis, a number of club cells were strongly stained. Because no agglutinating activity was detected in plasma, it appears evident that congerin is produced and secreted into mucus by those cells. In addition, congerin-positive club cells were distributed in the mucosal epithelium lining the digestive tract preceding the stomach and in the gills. These findings suggest that congerin participates in innate immunity on the intra- and the extra-body surface of the conger. The putative functions of club cells in fish and their contained lectin are discussed.
Animals↗
Purification and characterization of ostrich prothrombin.
The work focused on the penultimate enzyme, prothrombin, in the coagulation cascade. Prothrombin was purified and characterized from ostrich plasma. The results obtained contribute to a better understanding of blood coagulation in the ostrich and the evolution of prothrombin and the coagulation cascade. Prothrombin was purified from ostrich plasma by barium chloride precipitation, ammonium sulfate fractionation, and DEAE-cellulose and Cu(2+)-chelate Sepharose chromatography. Ostrich prothrombin exhibited a M(r) of 72,800 and a pI of 6.9 using SDS-PAGE and PAG-isoelectrofocusing, respectively. The N-terminal sequence of ostrich prothrombin showed 78 and 87% identity with human and bovine, respectively. The cDNA was isolated from ostrich liver and the predicted amino acid sequence compared with those from other species. Ostrich prothrombin shares sequence identity with chicken (84%), human (60%), bovine (59%), rat (60%), mouse (59%) and hagfish (50%) prothrombin, suggesting a common function of prothrombin in these vertebrates. Amino acid sequence identities indicate that the thrombin beta-chain (62%) and the propeptide-Gla (75%) domains are the regions most constrained for the common functions of vertebrate prothrombins. Ostrich prothrombin, therefore, shows similarity in structure to other vertebrate prothrombins.
Amino Acid Sequence↗
Hemodynamic changes with posture in calves with total artificial heart.
Hemodynamic changes with posture, sitting versus standing, were analyzed in five Holstein calves with the Cleveland Clinic-Nimbus TAH. This total artificial heart (TAH) has a left master alternate control mode that adjusts the pump rate and consequently pump flow proportional to the pulmonary venous return to the left pump (AUTO period). However, in this series of experiments, the pump reached its maximum beat rate within 1-5 days post operatively, after which pump flow could not increase (MAX period). Hemodynamic parameters (RAP, LAP, PAP, AoP, and pump flow) were obtained every 15-20 min throughout the experiments for as long as 120 days and averaged for each posture for each period. During the AUTO period, the flow while standing was significantly higher than that while sitting (standing: 8.7 +/- 0.2 L/min; sitting: 7.5 +/- 0.4 L/min; p < 0.05), and the systemic vascular resistance (SVR) was significantly lower (standing: 895 +/- 93 dyne.sec.m-5; sitting: 1,041 +/- 124 dyne.sec.m-5; p < 0.05). During the MAX period, the AoP and SVR standing were significantly lower than those sitting (AoP standing: 91 +/- 7 mmHg; AoP sitting: 98 +/- 7 mmHg; p < 0.05; SVR standing: 652 +/- 75 mmHg; SVR sitting: 730 +/- 96 mmHg; p < 0.05). The Cleveland Clinic-Nimbus TAH responded well to these changes in position, increasing pump flow and maintaining the AoP during the AUTO period.
Animals↗
The effects of radiation therapy on the tissue capsule of soft tissue implants.
A prosthesis has been developed for cosmesis after lumpectomy surgery for breast carcinoma. The device is saline filled and percutaneously adjustable in volume to permit an optimal cosmetic result after surgical wound healing. A series of 24 studies of 18 weeks' duration using the adult rabbit animal model were first used to study tissue capsule formation around textured versus smooth surface control implants and to evaluate the effects of volume adjustments on the tissue capsule. Single or multiple adjustments of implant volume had no effect on tissue capsule thickness or morphology. Because lumpectomy surgery is invariably followed by radiation therapy, a series of six studies was then conducted to determine the effects of a typical course of radiation therapy on tissue capsule formation. One week after device implantation, a 4 x 4 cm field including the implant was irradiated with 5,000 rad (200 rad/day x 5 days/week x 5 weeks). The animals were maintained for a 6 week period after radiation treatment. After sacrifice, the implants were removed, and the tissue capsules studied using conventional histologic techniques, including scanning and transmission electron microscopy. There was no statistically significant difference in tissue capsule thickness compared to nonirradiated controls. Tissue capsule morphology, however, differed markedly. Radiation therapy decreased angiogenesis, cellularity, and the inflammatory cell response to the implants. Qualitatively, radiation treatment seemingly improved rather than compromised the connective tissue response to the implants.
Animals↗
Adaptation of tissue to a chronic heat load.
Determination of the chronic effect of heat on tissue is one of the important issues facing mechanically actuated total artificial heart (TAH) development. In an effort to characterize this effect, implantations of heating devices producing constant heat fluxes of 0.04 watts/cm2, 0.06 W/cm2, and 0.08 W/cm2 were performed in 11 calves (H-series). Heated disks were implanted adjacent to lung and muscle tissue for a period of 7 weeks. Temperature sensors were placed at the surface as part of the heater assemblies. The results showed that initially, temperature elevations above body temperature (delta T) were 6.4 +/- 0.6 degrees C, 4.5 +/- 0.2 degrees C, and 1.8 +/- 0.5 degrees C at the muscle heater surface for 0.08, 0.06, and 0.04 W/cm2, respectively. At 2 weeks after implant delta T values changed to 5.5 +/- 0.6 degrees C, 3.4 +/- 0.2 degrees C, and 1.8 +/- 0.2 degrees C, respectively. Seven weeks after implant, delta T values decreased to 3.7 +/- 1.2 degrees C, 2.8 +/- 0.1 degrees C, and 0.8 degrees C for 0.08, 0.06, and 0.04 W/cm2, respectively. The authors think this change is attributable to an adaptive response of the tissue to increase heat dissipation through angiogenesis. Results from three TAH cases indicated that at two measured tissue interfaces, delta T decreased by 1 degrees C during a 15 day period. At the same time, the waste heat (volts x current in-flow x afterload to the blood) remained constant at 11.1 +/- 0.5 W during this period. This decrease in delta T corresponded to that observed for the H-series experiments at the higher heat fluxes. Thus, it appears that adaptation observed in the H-series experiments also is seen for tissues surrounding heat sources such as the TAH.
Adaptation, Physiological↗
Development of a magnetically operated artificial urethral sphincter. Chronic effects of compression on the skin structure and blood flow.
The artificial urethral sphincter (AUS) has been in clinical use for more than 20 years. Currently available AUS devices, however, are difficult to use and not entirely reliable. A magnetically operated AUS is currently under the development. Although the skin between the magnets will be compressed all day long, little information exists on the effects of chronic pressure on the skin structure and blood flow. In five miniature pigs, two internal magnets and one control metal disk were implanted subcutaneously at three different positions, and external magnets with differing magnetic forces were applied to the skin overlying the internal magnets for six weeks. In four pigs, the skin blood flow was measured by a laser Doppler flow meter applying different pressures. Compression of 10 mmHg preserved normal skin morphology in all but one animal where blood flow had not recovered 2 weeks after surgery. Compression of 20 mmHg for 6 weeks, however, produced pressure ulcers in all five cases (p < 0.05 vs. 10 mmHg group). The skin blood flow declined for pressures exceeding 20 mmHg (0 mmHg: 4.3 +/- 1.2, 10 mmHg: 4.3 +/- 3.3, 20 mmHg: 2.6 +/- 2.7 ml/min/100 g). We concluded that the magnetically operated AUS should use a magnetic coupling with a pressure less than 10 mmHg exerted on the interposing skin.
Animals↗
Effects of a total artificial heart right stroke volume limiter on left-right hemodynamic balance.
In a completely implantable total artificial heart (TAH), the left-right flow difference attributable to higher volumetric efficiency of the right pump and bronchial artery shunting has always been a significant problem. The automatic control of the Cleveland Clinic-Nimbus TAH accommodates for the left-right flow difference when the beat rate is below maximum (AUTO range). However, at its maximum beat rate (MAX), high left atrial pressure (LAP) (greater than 25 mmHg) with relatively low right atrial pressure (RAP) (less than 8 mmHg) were observed both in vitro and in vivo, suggesting the need for a stroke volume limiter (SVL) of the right pump. In vitro volume loading tests showed 10%, 15%, and 20% SVLs prevented a disproportionate increase of LAP. In vivo studies in five Holstein calves also showed a balanced LAP-RAP relationship with 10% and 15% SVLs at MAX. The left pump flow was not affected by this range of SVLs either in vitro or in vivo. Pulmonary function was maintained with either size SVL, with autopsies revealing absence of pulmonary congestion and minimal pleural effusions in two calves surviving for more than 1 month. Although additional studies are needed to determine the appropriate size of the SVL, both 10% and 15% SVLs were effective in maintaining left-right hemodynamic balance in this TAH.
Animals↗
Progress in Cleveland Clinic-Nimbus total artificial heart development.
A totally implantable, Cleveland Clinic-Nimbus total artificial heart (TAH) uses electrohydraulic energy conversion and an automatic left master-alternate mode control scheme, with a filling sensitivity of 1.0 l/min/mmHg and a maximum output of 9.5 l/min. The TAHs were tested in 12 calves for 1-120 days with normal major organ and blood cell function. Post-operative suppression of platelet aggregation recovered by the second post-operative week. The gelatin-coated pump surface generally was clean without any anticoagulants and free from infection. Embolism, which occurred in two cases, was caused by complications attributable to fungal infection in a Dacron graft and by thrombus formed around a jugular vein catheter. A system with a hybridized microcircuit controller in the interventricular space has been tested successfully in the three most recent cases, with a peak device surface temperature elevation of 6.5 degrees C. Heat effects were confined to the tissues immediately adjacent to the hottest spots. The carbon fiber-reinforced epoxy housing and 60 ml butyl rubber compliance chamber showed good tissue compatibility with a thin, fibrous tissue capsule. The transcutaneous energy transmission system and the internal battery functioned well as designed in the most recent animal implant.
Animals↗
Analysis of subcutaneous fluid in rats.
Subcutaneous implantation in rats is a commonly used model for biomaterial calcification studies. Although this model is frequently used, its components have not been characterized with respect to calcification. Exudate from the subcutaneous spaces of 18 young rats was collected using diffusion chambers. These chambers consisted of polymethylmethacrylate tubes with 0.22 micron pore filters covering each end allowing fluid, but not cells, to enter the chambers. Glutaraldehyde treated bovine pericardial strips were implanted subcutaneously, inside the chambers and outside the chambers, to test the calcification inducing abilities of the various environments. The animals were killed on postoperative day 10, and the exudate and materials were collected. The exudate was analyzed for ionic calcium, total calcium, inorganic phosphorus, and albumin, and for cells by a differentiated cell smear. The materials were analyzed for calcification by radiography, histology, and atomic absorption. Calcification was present in the materials inside the chambers where no cells were present and in the materials that were not in chambers. The distinct features of the exudate were elevated ionic calcium, a high Ca x P product, and elevated phosphorus.
Adult↗