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Biomedical subjects

M Maeda

Publications and source records attributed to M Maeda.

At least 775 records · Page 43Linked to original sources

Escherichia coli H(+)-ATPase: role of the delta subunit in binding Fl to the Fo sector.

The roles of the Escherichia coli H(+)-ATPase (FoFl) delta subunit (177 amino acid residues) was studied by analyzing mutants. The membranes of nonsense (Gln-23----end, Gln-29----end, Gln-74----end) and missense (Gly-150----Asp) mutants had very low ATPase activities, indicating that the delta subunit is essential for the binding of the Fl portion to Fo. The Gln-176----end mutant had essentially the same membrane-bound activity as the wild type, whereas in the Val-174----end mutant most of the ATPase activity was in the cytoplasm. Thus Val-174 (and possibly Leu-175 also) was essential for maintaining the structure of the subunit, whereas the two carboxyl terminal residues Gln-176 and Ser-177 were dispensable. Substitutions were introduced at various residues (Thr-11, Glu-26, Asp-30, Glu-42, Glu-82, Arg-85, Asp-144, Arg-154, Asp-161, Ser-163), including apparently conserved hydrophilic ones. The resulting mutants had essentially the same phenotypes as the wild type, indicating that these residues do not have any significant functional role(s). Analysis of mutations (Gly-150----Asp, Pro, or Ala) indicated that Gly-150 itself was not essential, but that the mutations might affect the structure of the subunit. These results suggest that the overall structure of the delta subunit is necessary, but that individual residues may not have strict functional roles.

Amino Acid Sequence↗

Assembly of a hybrid from the alpha subunit of Na+/K(+)-ATPase and the beta subunit of H+/K(+)-ATPase.

Messenger RNA for the alpha subunit of Torpedo californica Na+/K(+)-ATPase was injected into Xenopus oocytes together with that of the beta subunit of rabbit H+/K(+)-ATPase. The Na+/K(+)-ATPase alpha subunit was assembled in the microsomal membranes with the H+/K(+)-ATPase beta subunit, and became resistant to trypsin. These results suggest that the H+/K(+)-ATPase beta subunit facilitates the stable assembly of the Na+/K(+)-ATPase alpha subunit in microsomes.

Adenosine Triphosphatases↗

Genetic linkage between copper accumulation and hepatitis/hepatoma development in LEC rats.

The concentration of copper in the livers of Long-Evans rats with cinnamon-like coat color (LEC), in which hepatitis and then hepatomas develop spontaneously, was recently found to be abnormally high. Therefore, we examined the copper concentrations in the livers of LEC F1 backcrosses (LEC F1 x LEC) to determine the linkage of copper accumulation with development of hepatitis. Consistent with a previously reported ratio of rats with hepatitis to rats without hepatitis of about 1:1, hepatitis developed in 14 of 30 F1 backcrosses. The copper concentrations in the livers of all LEC F1 backcrosses with hepatitis were abnormally high and comparable to those of LEC rats. In contrast, the concentrations in all backcrosses without hepatitis were similar to those in normal Long-Evans with agouti coat color or Brown-Norway rats. Copper accumulation was shown to be closely linked with the development of hepatitis in LEC rats and appeared to be a possible cause of hepatitis. The concentrations of copper in the livers of Fischer 344 rats after carbon tetrachloride treatment were in the range for normal liver, indicating that a high copper concentration in the liver is specific to LEC rats and not a specific characteristic of hepatitis. Furthermore, we found that the size and level of ceruloplasmin mRNA in the livers of LEC rats were the same as those in LEA rats and that the size and level of ceruloplasmin polypeptide in their livers and plasma were almost the same as those in LEA rats. Therefore, these results suggest that the copper accumulation is not due to alteration of expression or to gross alteration of the ceruloplasmin gene.

Animals↗

Continuous local intraarterial infusion after prolonged arterial stasis in the fingers and toes.

Seven patients with trauma to eight digits and one toe went untreated for arterial stasis, with subsequent development of posttraumatic changes in skin coloration. In two patients involving two digits, a daily dose of 2,000 ml containing 240,000 U urokinase, 80 micrograms prostaglandin E1, and 10,000 U heparin in lactated Ringer's solution was administered by intravenous infusion for 10 consecutive days; one of the two digits became necrotic. In all subsequent patients, a daily dose of 80 ml containing 240,000 U urokinase, 40 micrograms prostaglandin E1, 10,000 U (maximum) heparin, and low-molecular-weight dextran was administered by continuous local intraarterial infusion for 10 consecutive days. These seven extremities survived, even in the case of two digits and one toe with over 50 hr of arterial stasis. We believe that revascularization of extremities following prolonged periods of arterial stasis may be possible by means of continuous local intraarterial infusion of antithrombotic agents.

Aged↗

Effectiveness of planar image and single photon emission tomography of thallium-201 compared with gallium-67 in patients with primary lung cancer.

A comparative study of planar images and single photon emission tomography (SPET) of thallium-201 chloride and gallium-67 citrate was performed in 38 patients with proven primary lung cancer to detect the primary lung tumour and to establish the presence of metastasis in the lung hilum and mediastinum. The findings of planar images and SPET were compared with the pathological findings after thoracotomy. It was shown that 201Tl studies were superior to 67Ga studies for evaluation of the primary lesion and lymph node metastases.

Aged↗

Intracerebral distribution of albumin after transient cerebral ischemia: light and electron microscopic immunocytochemical investigation.

The blood-brain barrier breaks down following cerebral ischemia, but the exact sequence of events for extravasation of serum proteins and their parenchymal distribution remain uncertain. We studied the distribution of serum albumin in the hippocampus of the gerbil brain using light and electron microscopic immunocytochemical techniques. With light microscopy, there was no reaction for albumin for the first 12 h after unilateral common carotid artery occlusion for 10 min and reperfusion. At 12 h, the reaction was weak and limited to the neuropil in the subiculum-CA1 region (between the subiculum and the medial CA1 region). After 24 h, the reaction became intense in the neuropil and neuronal perikarya in the subiculum-CA1 and medial CA1 regions. The electron microscopic immunocytochemical study of the subiculum-CA1 and medial CA1 regions revealed electron-dense immunoprecipitates in the extracellular space and the peripheral part of the apical dendrites as early as 30 min after reperfusion and in the astrocytic cytoplasm after reperfusion for 1 h. However, immunoprecipitates were not found in the neuronal perikarya until after reperfusion for 24 h. The present study demonstrated prompt appearance of albumin in the extracellular space of the brain parenchyma after re-establishment of cerebral circulation and prompt accumulation in the peripheral part of the dendrites with spreading to neuronal perikarya, likely in the process of degeneration and death.

Animals↗

Retrograde cerebral perfusion through a superior vena caval cannula protects the brain.

Retrograde cerebral perfusion through a superior vena caval cannula is a new technique for protecting the brain during aortic arch operations. In mongrel dogs (n = 10; 13 to 15 kg) we have performed retrograde cerebral perfusion (300 mL/min) by infusing blood through a superior vena caval cannula with aortic and inferior vena caval drainage. We have measured the cerebral tissue blood flow, oxygen consumption, and carbon dioxide exudation during retrograde cerebral perfusion at normothermia (NT, 37 degrees C) and hypothermia (HT, 20 degrees C) and have compared these values with values obtained in dogs during cardiopulmonary bypass (1,200 mL/min). Cerebral tissue blood flow was measured by the hydrogen clearance method. During retrograde cerebral perfusion about 20% of the superior vena caval perfusate was returned through the aorta and the rest drained from the inferior vena cava. Cerebral vascular resistance during retrograde cerebral perfusion was lower than that during cardiopulmonary bypass (NT, 63.8 +/- 52.5 versus 126.9 +/- 58.4; HT, 28.4 +/- 32.8 versus 69.5 +/- 28.7 x 10(3) dynes.s.cm(-5). Retrograde cerebral perfusion provided half the cerebral tissue blood flow of cardiopulmonary bypass (NT, 14.7 +/- 6.4 versus 34.3 +/- 7.8; HT, 17.6 +/- 5.6 versus 37.2 +/- 10.6 mL/min). Retrograde cerebral perfusion also provided a third of the oxygen (NT, 4.4 +/- 2.1 versus 12.3 +/- 7.1; HT, 1.4 +/- 0.8 versus 4.2 +/- 1.3 mL/min) and discharged 20% of the carbon dioxide (NT, 0.24 +/- 0.08 versus 1.19 +/- 0.58; HT, 0.15 +/- 0.06 versus 0.51 +/- 0.17 mmol/min) when compared with cardiopulmonary bypass. Retrograde cerebral perfusion may reduce ischemic damage during interruption of cerebral blood flow.

Animals↗

A study on optimal temperature for isolated lung preservation.

Fifty-two heart-lung blocks (grafts) of New Zealand white rabbits were used for determining optimal temperature in lung preservation. Grafts were inflated with room air and preserved by simple surface cooling at arbitrarily determined temperatures for 18 hours. Graft function was assessed by nonrecirculated perfusion with autologous blood. Segmented regression models between functional parameters and preservation temperature (T) were applied for determining optimal temperature. Graft ability was also assessed from the point of view of pulmonary circulation by indocyanine green dilution rate of effluent and histological distribution of carbon particles. Significant segmented regression curves and lines between parameters of effluent oxygen tension (PO2) and wet-dry weight ratio (W/D), and T were obtained as follows: PO2 = 150/(1 + 3208.1[e-1.17T]), p less than 0.01; PO2 = -13.8T + 222.6, p less than 0.05; W/D = 5.0 + 1.5/(1 + 0.0028[e0.94T]), p less than 0.01; W/D = 0.075T + 4.52, p less than 0.05. Optimal temperature for lung preservation by topical cooling was calculated as 8 degrees to 9 degrees C from each intersecting point of regression equations. Analysis of regression curves suggested that the most common hypothermic ischemic injury during preservation by topical cooling is pulmonary vascular obstruction, which might be induced at temperatures lower than the critical temperature of 6 degrees to 7 degrees C. Indocyanine green dilution rate and histological findings supported the results of graft functional parameters.

Animals↗

Role of the amino terminal region of the epsilon subunit of Escherichia coli H(+)-ATPase (F0F1).

Escherichia coli strain KF148(SD-) defective in translation of the uncC gene for the epsilon subunit of H(+)-ATPase could not support growth by oxidative phosphorylation due to lack of F1 binding to Fo (M. Kuki, T. Noumi, M. Maeda, A. Amemura, and M. Futai, 1988, J. Biol. Chem. 263, 17, 437-17, 442). Mutant uncC genes for epsilon subunits lacking different lengths from the amino terminus were constructed and introduced into strain KF148(SD-). F1 with an epsilon subunit lacking the 15 amino-terminal residues could bind to F0 in a functionally competent manner, indicating that these amino acid residues are not absolutely necessary for formation of a functional enzyme. However, mutant F1 in which the epsilon subunit lacked 16 amino-terminal residues showed defective coupling between ATP hydrolysis (synthesis) and H(+)-translocation, although the mutant F1 showed partial binding to Fo. These findings suggest that the epsilon subunit is essential for binding of F1 to F0 and for normal H(+)-translocation. Previously, Kuki et al. (cited above) reported that 60 residues were not necessary for a functional enzyme. However, the mutant with an epsilon subunit lacking 15 residues from the amino terminus and 4 residues from the carboxyl terminus was defective in oxidative phosphorylation, suggesting that both terminal regions affect the conformation of the region essential for a functional enzyme.

Amino Acid Sequence↗

The effect of mixed live vaccines of Newcastle disease and infectious bronchitis on the chicken respiratory tract.

The effect of mixed live vaccines of Newcastle disease (ND) and infectious bronchitis (IB) on specific pathogen-free chickens aged 7 days was investigated. The chickens were inoculated intranasally with mixed live vaccines with and without Escherichia coli, or with E. coli alone. "Vaccine 1" consisted of ND virus strain B1 and IB virus strain ON; "vaccine 2" consisted of ND virus strain B1 and IB virus strain H120. The tracheas of chickens inoculated with the vaccines and E. coli and with the vaccines alone showed multiplication of E. coli and histological lesions (loss of cilia, degeneration and hyperplasia in epithelial cells and cellular infiltration of subepithelial tissues). In the tracheas of chickens inoculated with vaccines and E. coli, multiplication of E. coli was greater than in chickens given vaccine alone. There were no histological lesions, and only mild, transient multiplication of E. coli in chickens inoculated with E. coli alone. The results suggest that these mixed live vaccines, especially vaccine 2, play a role in inducing or enhancing colibacillosis in the chicken.

Animals↗

Data processing for multi-channel optical recording: action potential detection by neural network.

Using a neural network, we have developed a program for fast and precise detection of action potentials (AP) in raw multi-channel optical recording data. The AP detection was performed in two steps: first, peaks were detected in raw optical data, and, second, the peaks were classified by the neural network into APs, noise and undecided peaks. The network was optimized and trained by the backpropagation learning algorithm, employing some thousands of manually classified peaks. The performance of the optimized network was found to be not completely satisfactory, although it was better than the classification by template matching and nearest-neighbor rules. The addition of a signal-to-noise ratio (SNR) of a peak to the network classification improved the classification performance: in comparison with the manual classification results, 96% of manually classified APs were detected. The causes of classification errors were discussed. In spite of the fact that the program required a slight amount of human intervention for undecided peaks, the program could allow mostly automatic AP detection.

Action Potentials↗

Sympathetic and metabolic mechanisms of the cerebrovasomotor function of the caudal ventrolateral medulla in rats.

We attempted to elucidate the cerebrovasomotor function of the caudal ventrolateral medulla. Sixty-one rats were anaesthetized, paralysed and artificially ventilated. The microsphere method was employed for the measurement of blood flow. Microinjection of an antagonist of excitatory amino acids, kynurenate (2 nmol), into functionally identified depressor sites within the caudal ventrolateral medulla produced arterial hypertension of about 140 mmHg. We found that the cerebral blood flow was substantially increased, but was maintained at the same level (17 rats) as that observed under phenylephrine-induced hypertension (26 rats). Bilateral severing of the cervical sympathetic trunks resulted in a further increase in blood flow in all brain regions studied (18 rats). The response was most significant in the cerebral parasagittal cortex (164 +/- 31% of baseline without, and 211 +/- 43% with sympathectomy; mean +/- S.D.; P < 0.001). The contributions of the cerebral metabolic mechanism to this flow increase under denervation was minimal, as evidenced by the observation of disproportionately smaller changes in cerebral metabolic rate for oxygen during any type of hypertension. We conclude that the cerebrovasomotor functions of the caudal ventrolateral medulla may operate to keep an equilibrium between simultaneously working tonic inhibitions against sympathetic vasoconstriction as well as against vasodilatation. This dual effect is mediated by excitatory amino acid receptors located within this particular brain area. The vasodilator mechanism may be of neurogenic origin. When the function of the brain area is suppressed, the subsequently disinhibited vasodilator mechanism dominates the cerebrovascular autoregulatory function.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Synthesis and biodistribution of a fluorine-18 labeled analogue of D-talose: 2-deoxy-2-[18F]fluoro-D-talose.

A fluorine-18 labeled analogue of D-talose, 2-deoxy-2-[18F]fluoro-D-talose ([18F]FDT), was synthesized via nucleophilic fluorination with [18F]fluoride ion and its biodistributions in animals were examined. Radiofluorination of benzyl 3,5,6-tri-O-benzyl-2-O-(trifluoromethanesulfonyl)-alpha-D-galac tof uranoside (5) with aminopolyether supported potassium [18F]fluoride (K18F/Kry222) in acetonitrile followed by deprotection of the [18F]fluorinated intermediate (6) with boron tribromide in CH2Cl2 gave [18F]FDT in an average radiochemical yield of 29% with a radiochemical purity greater than 98%. Biodistribution studies of [18F]FDT in mice bearing fibrosarcoma showed the highest uptake of radioactivity in the liver (34.9% dose/g), followed by the kidney (15.9%dose/g), the small intestine (12.9%dose/g) and fibrosarcoma (5.7%dose/g), at 30 min after i.v. administration. Although the radioactivity in the kidney and small intestine decreased with time, the uptake in the liver and the tumor slightly increased until 120 min. The high liver uptake of [18F]FDT was also observed in normal rats and this uptake was strongly inhibited by co-administration of D-galactose. These preliminary results suggest that [18F]FDT might be metabolized through the galactose metabolic pathway as analogously observed with 2-deoxy-2-[18F]fluoro-D-galactose which is an isomer with respect to carbon-2 of [18F]FDT, and that it may be another candidate for studying liver function by positron emission tomography.

Animals↗

Positron labeled antioxidants: synthesis and tissue biodistribution of 6-deoxy-6-[18F]fluoro-L-ascorbic acid.

A one-pot synthesis of 6-deoxy-6-[18F]fluoro-L-ascorbic acid (18F-DFA) has been developed via nucleophilic displacement of a cyclic sulfate with no-carrier-added [18F]fluoride ion. Isolated radiochemical yields of around 15% were obtained with radiochemical purity of over 99% after overall synthesis time of 90 min. Tissue distribution studies with 18F-DFA in rats showed high uptake of radioactivity in the adrenals, kidneys, liver and small intestine--organs known to have high concentrations of L-ascorbic acid. The slow and low uptake of radioactivity in the brain was observed between 10 and 120 min after i.v. injection. In vivo behavior of 18F-DFA in mice bearing 3-methylcholanthrene-induced fibrosarcoma demonstrated its ability to accumulate in the tumor.

Animals↗

In vitro comparison of the spiral Z stent and the Gianturco Z stent.

A new spiral Z stent is described, and its characteristics are compared in vitro with those of the modified Gianturco Z stent. The spiral Z stent has a more uniform expansile force throughout its effective length; is more compressible, thus allowing for use of a smaller introductory catheter; and is more stable than the modified Z stent. Flexibility of both stent types is similar. Advantages of the spiral Z stent promise to be beneficial for clinical use.

Equipment Design↗

Silicone-covered expandable metallic stents in the esophagus: an experimental study.

Modified silicone-covered Gianturco expandable metallic stents were placed in the normal esophagus of six young pigs. Following endoscopic examination, the stents were placed using endoscopic and fluoroscopic control. The animals were observed for eating behavior and weight gain. Stents appeared to be tolerated well based on these parameters. Three stents remained in position for the full study period, and three stents migrated into the stomach during the study. The pigs were sacrificed at four weeks and postmortem examination performed. Esophageal wall thickening and nodular inflammation were noted at the sites where the wire skirts penetrated the mucosa. Injury was limited to the region of the wire skirts and there was no injury due to the radial force of the stent body. There was no free perforation. This preliminary study suggests that endoscopic and fluoroscopic placement of modified silicone-covered Gianturco stents in the esophagus is feasible and safe. More extensive animal studies, followed by clinical investigation for palliation of malignant strictures, are warranted.

Animals↗