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Biomedical subjects

S Hashimoto

Publications and source records attributed to S Hashimoto.

At least 1,045 records · Page 58Linked to original sources

An improved technique for the closure of ventricular septal defect in ventricular inversion.

An improved technique for the closure of ventricular septal defect associated with ventricular inversion through the anatomically left ventricle is described. The technique consists of placing a patch completely on the anatomically right ventricular side with modified horizontal mattress sutures all around the defect. This technique was applied successfully in 2 patients with no ensuing conduction disturbance. The details of the technique are reported.

Adolescent↗

Multiple forms of immunoreactive renin in human adrenocortical tumour tissue from patients with primary aldosteronism.

There is increasing evidence which suggests that the adrenal gland contains the renin-angiotensin cycle. The localization of renin has been reported to be mainly in the zona glomerulosa rather than the fasciculata medullary portion. In the present study we have investigated extracts from aldosteronomas (n = 3), which are believed to derive from the zona glomerulosa cells. In addition, we have attempted to characterize the biochemical properties of the adrenal renin. Sizable quantities of renin-like activity (32.0 +/- 7.7 ng of angiotensin I generated h-1 mg-1 of protein, mean +/- SEM) were detected in the extracts. This renin-like activity was inhibited by anti-renin antibody raised against pure renin (mean, 95% of the total renin-like activity), indicating that it was not due to the non-specific action of proteases such as cathepsin D. The optimum pH of the tissue renin-like enzyme was 6.0 for rat plasma substrate. Differences were found, however, in the molecular mass (36,000, 37,000, 44,000 and 48,000), binding to concanavalin A and isoelectric points (4.40, 4.68 and 5.00). These results confirm the existence of specific renin in aldosteronoma. Renin microheterogeneity could be evidence for local production of the enzyme.

Adrenal Cortex Neoplasms↗

Sorting of three secretory proteins to distinct secretory granules in acidophilic cells of cow anterior pituitary.

The distribution of three proteins discharged by regulated exocytosis--growth hormone (GH), prolactin (PRL), and secretogranin II (SgII)--was investigated by double immunolabeling of ultrathin frozen sections in the acidophilic cells of the bovine pituitary. In mammotrophs, heavy PRL labeling was observed over secretory granule matrices (including the immature matrices at the trans Golgi surface) and also over Golgi cisternae. In contrast, in somatotrophs heavy GH labeling was restricted to the granule matrices; vesicles and tubules at the trans Golgi region showed some and the Golgi cisternae only sparse labeling. All somatotrophs and mammotrophs were heavily positive for GH and PRL, respectively, and were found to contain small amounts of the other hormone as well, which, however, was almost completely absent from granules, and was more concentrated in the Golgi complex, admixed with the predominant hormone. Mixed somatomammotrophs (approximately 26% of the acidophilic cells) were heavily positive for both GH and PRL. Although admixed within Golgi cisternae, the two hormones were stored separately within distinct granule types. A third type of granule was found to contain SgII. Spillage of small amounts of each of the three secretory proteins into granules containing predominantly another protein was common, but true intermixing (i.e., coexistence within single granules of comparable amounts of two proteins) was very rare. It is concluded that in the regulated pathway of acidophilic pituitary, cell mechanisms exist that cause sorting of the three secretory proteins investigated. Such mechanisms operate beyond the Golgi cisternae, possibly at the sites where condensation of secretion products into granule matrices takes place.

Animals↗

Discrimination and quantitative analysis of wild type and point mutant early region 1B genes of adenovirus using oligodeoxyribonucleotide hybridization probes.

One of the cytocidal (cyt) mutants of adenovirus 2 (Ad2), cyt15, has been shown to have two point mutations separated by 36 bases in its early region 1B (E1B) gene that encodes the 19K (Mr, 19,000) protein. A part of the cyt15 E1B gene, including one of the point mutations, was probed with a 23-base long oligodeoxyribonucleotide (ME1B23). Another oligonucleotide probe of the same length was used for the corresponding region of the wild type (wt) E1B gene (E1B23). Over the 32P-end labeled oligonucleotide probe, a 25-fold molar excess of a competitor (1) (the unlabeled oligonucleotide of each counterpart) was added to the hybridization mixture. The E1B gene with or without a point mutation could easily be distinguished from its counterpart and quantitated in the presence of its counterpart under the hybridization conditions employed. Under the stringent wash conditions, the E1B gene of wt Ad2 could be completely discriminated from even a 100-fold molar excess of cyt15 E1B gene with the 32P-labeled E1B23 probe. The wt E1B gene could also be quantitated in the presence of a 100-fold molar excess of the mutant E1B gene.

Adenoviridae↗

Identification of the principal promoter sequence of the c-H-ras transforming oncogene: deletion analysis of the 5'-flanking region by focus formation assay.

A number of deletion mutants were isolated, including 5', 3', and internal deletions in the 5'-flanking region of the human cellular oncogene related to the Harvey sarcoma virus (c-H-ras), and their transforming activities were examined in NIH 3T3 cells. DNA sequences which could not be detected without losing transforming activity were localized to a relatively short stretch upstream of the region which showed homology to the 5'-flanking region of v-H-ras oncogene. S1 nuclease analysis indicated that there were two clusters of mRNA start sites at positions that were about 1,371 and 1,298 base pairs upstream of the first coding ATG. The minimum region required for promoter function was estimated to be a 51-base-pair-long (or less) DNA segment. The promoter was GC rich (78%) and did not contain the consensus sequences that are usually observed in PolII-directed promoters but contained a GC box within which one of the mRNA start sites was included. In addition, two sets of positive and negative elements seemed to be located between the promoter and the protein-coding region, which appeared to influence positively and negatively, respectively, the efficiency of transformation with the c-H-ras oncogene.

Animals↗

Measurement of aortic blood flow with MR imaging: comparative study with Doppler US.

An innovative magnetic resonance imaging technique was applied to the measurement of blood flow in the abdominal aorta. The technique combines selective excitation and visualization from an orthogonal view. The distance that fluid has moved is directly visualized. The blood flow velocity at every 50 msec throughout the cardiac cycle was measured in a short time (about 4 minutes) using electrocardiographic gating and repeated excitations in each cycle. Measurements were compared with those obtained by Doppler ultrasound (US) as a reference. The pulsatile change of flow velocity in the cycle correlated well with the Doppler US recording. Two flow velocity indexes, peak flow velocity and the velocity integral, also showed good correlation (r = .98 for both). This method is applicable for clinical use and is useful for measurement of high flow rates, as found in arteries.

Adult↗

Subcellular redistribution of lysosomal enzymes during caerulein-induced pancreatitis.

The subcellular distribution of the lysosomal enzymes cathepsin B and D in the pancreas was evaluated in rats infused with saline (control) or a maximal (0.25 microgram . kg-1 . h-1) or a supramaximally stimulating dose (5 micrograms . kg-1 . h-1) of the secretagogue caerulein. The latter results in acute edematous pancreatitis, inhibition of digestive enzyme secretion, and the localization of digestive zymogens in organelles whose fragility has been increased by caerulein infusion [A. Saluja et al. Am. J. Physiol. 249 (gastrointest. Liver Physiol. 12): G702-G710, 1985]. Samples from control animals were found to have 29.9 +/- 1.8% of the cathepsin B activity in the pellet centrifuged at 1,300 g for 15 min (containing primarily zymogen granules) and 54.7 +/- 2.5% in the pellet centrifuged at 12,000 g for 12 min (containing primarily lysosomes and mitochondria). After supramaximal stimulation with caerulein for 3.5 h the pellet centrifuged at 1,300 g for 15 min had 55.1 +/- 2.5%, and the pellet centrifuged at 12,000 g for 12 min had 30.6 +/- 2.0% of cathepsin B activity. This redistribution was time dependent, noted within 1 h of starting caerulein infusion, and maximal after 2.5 h of infusion. Electron microscopic immunolabeling studies revealed localization of cathepsin D in discrete organelles that, in the samples from animals infused with a supramaximally stimulating dose of caerulein, were larger, more abundant, and more concentrated in the pellet centrifuged at 1,300 g for 15 min than in the controls. During infusion with supramaximal doses of caerulein, the cathepsin B-containing organelles were found to become progressively more fragile.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Intracellular transport of pancreatic zymogens during caerulein supramaximal stimulation.

Rats infused with a dose of the secretagogue caerulein that is in excess of that which stimulates a maximal rate of pancreatic digestive enzyme secretion develop acute edematous pancreatitis. We have previously noted that infusion of this dose of caerulein (5 micrograms . kg-1 . h-1) induces the appearance of large heterogeneous vacuoles in acinar cell, blockade of exocytosis, and intracellular accumulation of digestive zymogens [O. Watanabe et al. Am. J. Physiol. 246 (Gastrointest. Liver Physiol. 9): G457-G467, 1984 and A. Saluja et al. Am. J. Physiol. 249 (Gastrointest. Liver Physiol. 12): G702-G710, 1985]. The current studies were performed to further elucidate these phenomena at the electron microscopic level of resolution and employed the techniques of pulse labeling, radioautography, and immunolocalization. Rats were infused with caerulein (5 micrograms . kg-1 . h-1) for 1 h, given a pulse of [3H]phenylalanine, and killed at selected times during the subsequent 5- to 180-min postpulse period during which caerulein infusion was continued. Transport from the endoplasmic reticulum to the Golgi cisternae was not altered by supramaximal stimulation, but transport through post-Golgi elements was altered. In particular, the maturation of condensing vacuoles into zymogen granules was found to be impaired. This led to the accumulation of partially condensed vacuoles and to the development of the large vacuoles containing newly synthesized digestive zymogens as well as the lysosomal hydrolase cathepsin D. The source of the latter could be impaired sorting of lysosomal and digestive enzymes and/or fusion of vacuoles with lysosomes. At the later times after pulse labeling, mature zymogen granules were also found to fuse with these large cathepsin D-containing vacuoles by a process analogous to crinophagy. Thus these studies indicate that the large heterogeneous vacuoles that appear during supramaximal secretagogue stimulation and that contain admixed digestive zymogens and lysosomal hydrolases arise by at least two mechanisms, impaired condensing vacuole maturation and crinophagy.

Animals↗

Biochemical characterization of angiotensin-converting enzyme in human neuroblastoma tissue.

High activity of angiotensin-converting enzyme was demonstrated in human neuroblastoma tissue. This activity required the presence of chloride ion and was almost completely inhibited by a specific converting enzyme inhibitor captopril (10 nM), indicating that the activity measured is indeed angiotensin-converting enzyme. Furthermore, the biochemical features of the enzyme were closely similar to the well-known properties of human lung converting enzyme, such as molecular weight (290,000), optimum pH (8.0-8.5), the presence of glycoprotein residues, and dependence on chloride ion concentration. These results provide definitive evidence for the presence of true angiotensin-converting enzyme in human neuroblastoma tissue.

Angiotensin-Converting Enzyme Inhibitors↗

[Experimental considerations of myocardial single photon emission CT image with a cardiac phantom].

We studied the quantity of thallium-201 myocardial single photon emission CT (SPECT) image with a cardiac phantom. The myocardial SPECT image is influenced by several causes, i.e., the absorption of gamma rays, collimator aperture, statistical noise and limited angle of rotation in data acquisition. The attenuation correction under the assumption of uniform distribution is not sufficient, so we have to consider the actual distribution of absorption. The effects of the nonuniform attenuation are most prominently appeared when we evaluate the quantity by the integral method. The aperture characteristics of a collimator are decided by the distance between the rotational center of the gamma camera and the collimator surface. Simultaneously scattered gamma rays affect the reconstructed image in the low frequency component, so the effective attenuation coefficient varies gradually. The statistical noise intermixed in the projection data generate artifacts like as lump-shaped pattern. The noise are clearly appeared in the spatial frequency upper than 0.25 cycle, so the low-pass filter are required with that of cut-off frequency. Moreover to enhance the details of the radionuclide distribution of cardiac muscle, the Wiener filter should be applied. The scan area also affects the appearance of the artifact in the myocardial SPECT image. The 180-degree scan usually generates false defects just nearby the center of reconstructed cross section. And the positions are changed with the conditions of gamma ray absorber and its relative position to the myocardial area. The 360-degree scan should be used from the standpoint of reconstructing more quantitative image.

Gamma Rays↗

Immunohistochemical change of actin in experimental myocardial ischemia. Its usefulness to detect very early myocardial damages.

Pathomorphological diagnosis of acute myocardial infarction has many problems in human autopsy materials less than 4 to 6 hours after clinical onset and in rats 2 to 3 hours after experimental coronary occlusion. Since immunohistochemical reaction depends on the antigen determinant site of the material, changes in the reaction may reflect alterations at the molecular level in myocardial fibers. With this consideration in mind, the effectiveness of diagnosing infarction at the earliest (possible) stage, and the changes of actin filaments were investigated through experiments, using immunohistochemical methods involving anti-actin antibodies produced from chicken gizzards in our laboratory. The left coronary arteries of rats were ligated to produce ischemia. Dehydrogenases were shown to be still present by triphenyltetrazolium chloride (TTC) reaction, but the anti-actin antibody reaction had disappeared in areas corresponding to the ischemic sites. However, on electron microscopic examination of these sites, actin fibers were clearly revealed. In the case of ischemia lasting for more than 6 hours, the anti-actin antibody reaction had disappeared, corresponding to the disappearance of the TTC reaction. At this stage, myocardial actin fibers were revealed by electron microscopic examination. These results indicate that ischemia induces some type of biochemical degeneration at the molecular level of myocardial actin, most likely the change of actin polymerization. Moreover, they show that the anti-actin antibody technique is capable of detecting such very early degenerative and ischemic changes proving itself to be better suited for determining the range and degree of early infarction.

Actins↗