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Biomedical subjects

K Murakami

Publications and source records attributed to K Murakami.

At least 883 records · Page 49Linked to original sources

Effect of fatty-acyl-CoAs on the elongation of saturated fatty acid in porcine aorta microsomes.

The microsomal elongation system from porcine aorta for longchain fatty-acyl-CoAs was investigated. Palmitoleoyl-CoA (16:1-CoA), oleoyl-CoA (18:1-CoA), and eicosenoyl-CoA (20:1-CoA) remarkably depressed the elongation activity for 16:0-CoA in aorta microsomes by 44.8, 52.4, and 43.7% of the control activity, respectively. Saturated and polyunsaturated fatty-acyl-CoAs had little effect on the 16:0-CoA elongation activity. These results indicate that monounsaturated long-chain fatty acyl-CoAs can regulate the synthesis of saturated fatty acids in the vessel walls.

Acyl Coenzyme A↗

Dietary cis-fatty acids that increase protein F1 phosphorylation enhance spatial memory.

Activation of protein kinase C (PKC) facilitates long-term potentiation (LTP), a model of memory, and increases its substrate protein F1 (aka GAP43) phosphorylation in direct relation to synaptic enhancement. Unsaturated fatty acids (c-FAs) which activate purified PKC, when injected into hippocampus, enhance LTP. To determine if dietary c-FAs could alter memory itself as well as brain PKC substrate (F1) metabolism, rats were maintained for 10 weeks on fatty acid diets enriched in mono-unsaturated oleic acid (OA; 20% olive oil, w/w), or a mono- and di-unsaturated mixture of oleate/linoleate (O/L; 20% corn oil), or a saturated fatty acid diet of laurate/myristate (L/M; 20% hydrogenated coconut oil). The O/L diet group was superior to the OA and L/M groups in spatial memory performance after the first two weeks of acquisition and in later achievement of criterion performance. The O/L diet had a significantly higher hippocampal protein F1 in vitro phosphorylation than in both the OA and L/M in trained and non-trained animals. Significantly, animals that made fewer errors showed higher F1 phosphorylation (r = -0.70). Diet both increases brain PKC substrate phosphorylation and enhances maze learning, confirming the feasibility of enhancing learning and memory by dietary regimens derived from basic neurochemical studies of synaptic plasticity.

Animals↗

Expression of a deletion mutant of the prosegment of human prorenin in Chinese hamster ovary cells.

Expression plasmids encoding native human preporenin and a mutant deleted in its entire prosegment were transfected into Chinese hamster ovary cells. The cells transfected with the expression plasmid of native preporenin secreted exclusively inactive prorenin, while the cells transfected with the mutant secreted the active enzyme. The secreted amount of renin from the latter cells was much lower than that of prorenin from the former ones, although these two enzymes had little difference in specific activity after trypsin activation. These results suggest that the prosegment plays an important role in the secretory process of renin, although the fully active enzyme can be formed in its absence.

Amino Acid Sequence↗

Tissue-specific expression of the human renin gene in transgenic mice.

Transgenic mice carrying human renin gene were produced by microinjection of 15 kilobases (kb) DNA molecules with up to 3 kb of 5'-flanking sequence and 1.2 kb of 3'-flanking sequence. The transgenes have been shown to be stably transmitted to progeny. It was revealed by RNase protection assay that the human renin gene in a transgenic mouse is expressed preferentially in the kidney. The human renin RNA was also detected at a small level in a variety of tissues such as brain, heart, lung, pancreas, spleen, stomach, testis, and thymus. The direct radioimmunoassay using a monoclonal antibody specific for the active site of human renin demonstrated the synthesis of human active renin in the transgenic mouse kidney. These results suggest that the human renin gene in the transgenic mouse is regulated in a tissue-specific manner.

Animals↗

[Effectiveness of pharmacoangiography using nitroglycerin and angiotensin II].

An investigation of pharmacoangiography using a combination of nitroglycerin and angiotensin II demonstrated that it was useful due to the following reasons. First, good dilatation of the proximal portion of the arteries was demonstrated which enabled good clarity of the margin between the encasement and the normal portion of the arteries. Secondly, there was almost no change demonstrated in blood pressure immediately after administration. Thirdly, tumor vessels and intrapancreatic arteries were well recognized on the resulting images.

Angiography↗

Two different forms of endothelin receptors in rat lung.

We provided evidence for the presence of two distinct types of rat endothelin (ET) receptors having different molecular masses by affinity labelling of rat lung membranes with 125I-ET-1, ET-2 and ET-3 and SDS-PAGE followed by autoradiography. One type, which is 44 kDa has a higher affinity for ET-1 and -2 than ET-3, whereas the other type, which is 32 kDa, preferably interacts with ET-3 rather than ET-1 and -2.

Affinity Labels↗

A cytocidal tissue kallikrein isolated from mouse submandibular glands.

A cytocidal factor against mouse thymocytes was purified from the submandibular glands of female BALB/c mice using Sephadex G-50 gel filtration chromatography and reverse-phase HPLC. SDS-PAGE and amino acid sequence analysis revealed that the cytocidal factor was mouse glandular kallikrein (mGK)-6. mGK-6 showed an optimal enzyme activity at pH 10 and a cytocidal activity against thymocytes in a dose-dependent manner.

Amino Acid Sequence↗

Prorenin is sorted into the regulated secretory pathway independent of its processing to renin in mouse pituitary AtT-20 cells.

A native human prorenin and a mutant of its processing site (Arg-43 to Gln) were expressed in mouse pituitary AtT-20 cells which process prorenin to renin and have both regulated and constitutive secretory pathways. The native prorenin was processed to renin and secreted in a regulated manner. Although the mutant precursor was not processed, it was also secreted in a regulated manner. These results suggest that prorenin is sorted into the regulated pathway, stored in secretory granules and released by stimulus whether it is processed to renin or not.

Amino Acid Sequence↗

Proteinic prorenin-releasing-stimulator (PRS) in the rat submandibular gland.

The release of prorenin as well as renin from rat renal slices was confirmed by a rat prorenin-prosegment ELISA system and an assay system for determining the renin activity. A significant increase of the prorenin release was found by adding rat submandibular gland extract to the slice medium, indicating the existence of a prorenin-releasing stimulator (PRS) in the extract. The pI and molecular mass of PRS were 8.5-8.7 and 28-30 kDa, respectively. The PRS was completely inactivated by boiling or a proteinase treatment.

Animals↗

Two distinct types of endothelin receptors are present on chick cardiac membranes.

Competitive displacement experiments of 125I-endothelin (ET)-1, -2, or -3 binding to chick cardiac membranes were performed with unlabeled ET-1, -2, -3, and sarafotoxin S6b (STX) as competitors. 125I-ET-1 and -2 binding was competitively inhibited by increasing concentrations of these unlabeled peptides in the same order; i.e. ET-2 greater than or equal to ET-1 greater than ET-3 greater than STX. In contrast, the order of potency in displacing 125I-ET-3 binding was ET-3 greater than ET-2 greater than or equal to ET-1 greater than STX. Affinity labeling of the membranes by cross-linking with 125I-ET-1 and -2 via disuccinimidyl tartarate yielded one major specific band with an apparent Mr = 53,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by autoradiography. On the other hand, affinity labeling with 125I-ET-3 showed that two major and one minor bands of Mr = 34,000, 46,000, and 53,000, respectively, were specifically labeled. These results indicate the presence of two distinct types of ET receptors, one of which has higher affinity for ET-1 and -2 than ET-3 and the other is conversely ET-3-preferring.

Animals↗

Renin expression in the kidney and brain is reciprocally controlled by captopril.

The tissue distribution of rat renin mRNA was examined. Sensitive RNase protection analyses demonstrated that renin mRNA are produced by the extra-renal tissues such as adrenal, brain, liver, lung, pituitary and testis. In response to sodium depletion and captopril treatment, the expression of mRNAs encoding rat renin were in a tissue-specific manner. The level of kidney renin mRNA remarkably increased in sodium-depleted rats treated with captopril, whereas that of brain renin mRNA definitely decreased. No significant change in the level of liver renin mRNA was observed after the same treatment. These results suggest that the expression of cerebral renin is regulated by physiological stimuli independent of its extra-cerebral expression.

Animals↗

Decrease of the inhibition of lipid peroxidation by glutathione-dependent system in erythrocytes of non-insulin dependent diabetics.

The inhibition of lipid peroxidation of erythrocyte membranes by glutathione-dependent protection was studied in patients with non-insulin dependent diabetes mellitus. Incubation of red cells from diabetics with 1.5 mM t-butyl hydroperoxide resulted in a lipid peroxidation increase greater than that of normal controls. Glutathione-dependent and glutathione-independent protection against oxidative damage was examined using an artificial system, in which erythrocyte ghosts were incubated with t-butyl peroxide and dialysed hemolysate in the presence or the absence of 2 mM glutathione. The glutathione-dependent protection of hemolysate from diabetics was approximately 70% of that from normal controls. The results suggest that decrease in glutathione-dependent protection against lipid peroxidation, along with decrease in glutathione levels, increases oxidative damage in erythrocyte membranes taken from diabetic patients.

Adult↗

[Quantitative evaluation of bone mineral contents of hand bone using CR technique].

We evaluated the clinical applicability of a matrix compression technique on computed radiography (CR) data for quantitative measurements of bone mineral content of hand bones. Similar results were obtained compared to those obtained by the conventional microdensitometry (MD) technique. This CR technique which uses imaging plates should take the place of the normal MD technique due to the fact that it provides for the reduction of exposure dosage.

Absorptiometry, Photon↗

Tissue distribution and characterization of prorenin-converting enzyme in mouse.

Renin is produced from an inactive precursor, prorenin, through endoproteolytic cleavage at paired basic amino-acid residues. Using (35S)methionine-labeled prorenin, that was synthesized with Xenopus oocyte expression system, as a substrate, we have determined the tissue distribution and the nature of prorenin-converting activity in mouse. The highest activity was found in the submandibular gland of male ICR mouse. The activity of the enzyme seemed to be parallel to that of renin. This enzyme activity, with an optimal pH 8.0-8.5, was inhibited by leupeptin, antipain and benzamidine.

Animals↗

[Improved contrast enhancement with a positive pressure drip infusion method and 350 mg I/ml high concentration contrast media in routine contrast enhanced CT].

Sixty cases of routine contrast enhanced CT were analyzed about the utilities of positive pressure drip infusion method using high concentration contrast media 350 mgI/ml, 100 ml. They were divided into three groups and each twenty cases were received positive pressure drip infusion method using Iohexol 350 mg/ml, 100 ml (350PP), positive pressure drip infusion method using Iohexol 300 mgI/ml, 100 ml (300PP) or conventional drip infusion method using Iohexol 300 mgI/ml, 100 ml (300DI). Enhancement effects of the liver parenchyma, the aorta and the spleen at upper, middle and lower levels of the liver were evaluated with increased attenuation on pre- and postcontrast CT. And intrahepatic contrast was evaluated with attenuation difference between liver parenchyma and intrahepatic vessels on postcontrast CT. In result, 300PP kept better enhancement effects and obtained better intrahepatic contrast than 300DI at the each level of the liver, and 350PP was still better than 300PP. The analysis of relationship between intrahepatic contrast and body weight suggested that 300PP was the optimal choice for the cases with 40-50 kg body weight and 350PP was for the ones with 50-60 kg as the infusion method of routine contrast enhanced CT. And the dose of contrast media seemed to be more needed for the cases over 60 kg body weight. 350PP and 300PP were concluded as useful, safe and simple methods for routine contrast enhanced CT of the liver.

Body Weight↗

A full length cDNA clone for the alkaline protease from Aspergillus oryzae: structural analysis and expression in Saccharomyces cerevisiae.

We have cloned and determined the nucleotide sequence of a cDNA fragment for the entire coding region of the alkaline protease (Alp) from a filamentous ascomycete Aspergillus oryzae. According to the deduced amino acid sequence, Alp has a putative prepro region of 121 amino acids preceding the mature region, which consists of 282 amino acids. A consensus sequence of a signal peptide consisting of 21 amino acids is found at the N-terminus of the prepro region. The primary structure of the mature region shares extensive homology (29%-44%) with those of subtilisin families, and the three residues (Asp 32, His 64 and Ser 221 in subtilisin BPN') composing the active site are preserved. The entire cDNA, coding for prepro Alp, when introduced into the yeast Saccharomyces cerevisiae, directed the secretion of enzymatically active Alp into the culture medium, with its N-terminus and specific activity identical to native Aspergillus Alp.

Amino Acid Sequence↗