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Biomedical subjects

K Miyazaki

Publications and source records attributed to K Miyazaki.

At least 739 records · Page 41Linked to original sources

Regulation by human chorionic gonadotropin of protein kinases in rabbit endometrium.

The effect of human chorionic gonadotropin (CG) on protein kinase levels has been studied in rabbit endometrium. A good correlation was observed between alterations of total protein kinase activity and DNA content in the tissue. The level of type I cyclic AMP-dependent protein kinase had slightly increased by 6 h after human CG treatment, and then decreased later. In contrast, the type II enzyme gradually increased after 3 days. The increase of the activity at 7 days was mainly due to that of type II enzyme, and the ratio of type II to type I enzyme increased. The activity pattern at the later stage closely resembled that of treatment with progesterone.

Animals↗

Purification and characterization of alkaline protease and neutral protease from chromatin of rats.

It was previously reported that, in addition to a known chymotrypsin-like protease capable of hydrolyzing histones with an optimum pH of 8 (neutral protease), another protease is bound to the chromatin of various rat tissues and in situ hydrolyzes casein more quickly than histones with an optimum pH of 10 (alkaline protease). In the present study, the alkaline protease was purified 14 000-fold to approx. 75% purity from the chromatin of Rhodamine sarcoma. This tumor contains both proteases at higher levels than normal tissues. For purification, affinity columns of Sepharose with bound soybean trypsin inhibitor, casein and histones were successively used. Also, the neutral protease was purified 920-fold to an apparently homogeneous state by affinity chromatography on a Sepharose column with bound soybean trypsin inhibitor under conditions, in which an excess amount of the enzyme was applied on the column so that part of the enzyme would pass through the column without adsorption and the enzyme thus adsorbed was then eluted. The purified alkaline and neutral proteases had molecular weights of approx. 18 000 and 27 000, respectively, and isoelectric points of approx. 11. The former enzyme hydrolyzed casein (100) in preference to histones (18) with an optimum pH of 9.5, whereas the latter enzyme preferred histones (100) to casein (32) with an optimum pH of 8. Their actions against other proteins and synthetic substrates were also studied.

Animals↗

Isolation and primary culture of adult human hepatocytes. Ultrastructural and functional studies.

Biopsy tissue of adult human liver was gently dissociated with collagenase followed by Dispase. By repeated low g centrifugation, a large number of almost pure, viable hepatocytes was obtained. This is the first report of a successful procedure for obtaining adult human hepatocytes for study in tissue culture. The isolated cells have the typical morphology of liver parenchyma, and these characteristics persist throughout the period of culturing. Evidence of their function is indicated by albumin synthesis. This procedure is now being used to study human hepatocyte functions in vitro and the effects of a variety of agents including carcinogens and viruses.

Adult↗

Peripheral and central vestibular disorders in alcoholics. A three-year follow-up study.

Vestibular disorders were found in all 33 Japanese men who had been drinking alcohol for over 20 years and who had abstained from alcohol for over 3 months before the tests were done. Six, 12, and 15 patients showed peripheral, central, and combined types of vestibular disorders, respectively. The peripheral disorder occurred in younger alcoholics (under 50 years old) with a drinking history of 20-30 years, and the combined disorder in older patients with a longer drinking history. The vestibular disorders, however, were not observed in any of 16 control non-alcoholics. A follow-up study on seven alcoholics with a 3-year abstinence from alcohol revealed an improvement of vestibular dysfunctions, in particular of peripheral disorders, with an abstinence of over 2 years.

Adult↗

Effect of furosemide in congestive heart failure.

The diuretic effect of furosemide was studied in 18 patients with congestive heart failure. Subjects were divided into two groups, group I consisting of eight patients with moderate and group II of 10 patients with advanced congestive heart failure. Six hours after bolus injection of furosemide (40 mg), mean urinary sodium was 120.5 +/- 36.7 mEq in group I and 68.2 +/- 25.8 mEq in group II (p less than 0.01), mean urine volume was 1,100 +/- 281 and 764 +/- 257 ml (p less than 0.05), mean urinary furosemide excretion was 28.08 +/- 2.60 and 24.00 +/- 0.74 mg (p less than 0.05), and mean furosemide renal clearance was 73.4 +/- 16.6 and 42.3 +/- 11.5 ml/min (p less than 0.001). Diuretic effect and furosemide renal clearance, as well as urinary furosemide excretion, correlated positively. The diuretic effect of furosemide with and without hydralazine (0.2 mg/kg) was compared in eight patients in group II. Urinary sodium excretion 6 hr after furosemide rose from 77.2 +/- 31.0 to 122.8 +/- 42.5 mEq after furosemide with hydralazine (p less than 0.01). Urine volume rose from 854 +/- 278 to 1,279 +/- 359 ml (p less than 0.001), urinary furosemide excretion rose from 23.64 +/- 2.03 to 26.94 +/- 2.30 mg (p less than 0.01), and furosemide renal clearance rose from 46.3 +/- 12.2 to 62.5 +/- 18.6 ml/min (p less than 0.01).

Adult↗

Chemotaxis of Kupffer cells isolated from rodent liver.

Kupffer cells (KC) were isolated from the liver of guinea pigs, rats, and mice using enzymatic digestion with collagenase, followed by differential centrifugation and plastic adherence. Purity of the isolated KC was 96.0 +/- 2.2, 97.2 +/- 2.1, and 96.0 +/- 2.3 per cent in guinea pigs, rats, and mice respectively. These isolated KC were tested for migratory response to bacterial factor, which is one of the representative chemotactic factors for inflammatory macrophages, using a modified Boyden chamber technique. KC from the three animal species similarly migrated to the bacterial factor. The migratory response of the KC to the bacterial factor is due to chemotaxis but not chemokinesis. These results show the possibility that KC may recognize a chemoattractant and directionally migrate to it.

Animals↗

Cyclic peptides. X. Bitter taste and chymotryptic hydrolysis of cyclic depsidipeptides containing a tryptophan residue.

Four stereoisomers (I-LL, I-LD, I-DL and I-DD) of a cyclic depsidipeptide (I) containing a tryptophan and a 2-hydroxy-4-methylpentanoic acid residue were synthesized, and their taste and chymotryptic susceptibility were examined. Compound I-LL is a depsipeptide analog of a bitter principle BP-II, cyclo(-L-Trp-L-Leu-), obtained from casein hydrolyzate. All of the four stereoisomers of I are strongly bitter to taste. Another depsipeptide analog, L-aspartyl-L-2-hydroxy-3-phenylpropanoic acid methyl ester, of the sweet H-L-Asp-L-Phe-OMe showed bitter taste instead of sweet. Chymotrypsin hydrolyzed I-LL and I-LD in moderate rates, and I-DL and I-DD very slowly.

Chymotrypsin↗

The behavior of pentaerythritol tetranicotinate in rat gastrointestinal tract as a prodrug.

The gas chromatographic assay method for pentaerythritol tetranicotinate, a nicotinic acid prodrug, and its hydrolysates was developed. The behavior of the drug in gastrointestinal tract was investigated in rat by using the method. The disappearance and hydrolysis of the drug were not observed in the gastric loop until 30 min. The rate of disappearance from the intestinal loop was 36.7% at 30 min which was significantly smaller than that of nicotinic acid. Little hydrolysis of the drug was observed in the buffer solution, pH 7.4, at 37 degrees up to 2 hr. However, the consecutive hydrolysis was observed when the drug was incubated with everted intestine or plasma. As to the rate of hydrolysis of the drug and its esterform hydrolysates by scraped intestinal mucosa, the ester to which the larger number of nicotinic acid was bound was hydrolyzed more rapidly. These results indicate that the orally administered drug is enzymatically hydrolyzed in the intestinal mucosa by a consecutive reaction. Although the hydrolysis rate of pentaerythritol tetranicotinate is rapid, the rate of its ester-form hydrolysate becomes slower gradually as the nicotinic acid is released. The released nicotinic acid is rapidly absorbed. The behavior of the drug revealed in this study suggests that pentaerythritol tetranicotinate is useful as a prodrug of nicotinic acid.

Animals↗

Survival of Salmonella typhi in oysters.

Shell oysters were reared in experimental conditions to elucidate the survival rate of Salmonella typhi in the body of cultured oysters and the fate of S. typhi in the shucked oysters. Sixty to 80 cultured oysters were reared for 4 h at 15 degrees C in 40 l of sea water than contained 1 x 10(4) -- 2 x 10(4) cells/ml of S. typhi (V-form) added, then these contaminated oysters were transferred to a tank that contained 40 l of clean sea water circulated through a ultraviolet disinfectant apparatus with a filtration system, and oysters were reared therein for 7-9 d at 15 degrees C. During the time the oysters were in the clean sea water, 5 oysters were periodically sampled, shucked, and the number of viable cells of S. typhi in the oysters was determined. At the beginning of the rearing in the clean sea water, the number was 4 x 10(5) -- 8 x 10(5) cells/100 g (MPN) in the shucked oysters. The MPN gradually lowered thereafter during the first 4 h in the clean sea water, but 24 h later, the MPN was reduced as much as 4 log. Thereafter, 20-80 cells/100 g (MPN) or 2-12 cells/oyster of S. typhi were enumerated up to the end of the experiment and a complete elimination was not observed. When the contaminated oysters were preserved at 10 degrees C, 5 degrees C or -20 degrees C, as shucked oysters, he number of viable cells of S. typhi did not change beyond the range of 1 log which may be caused by individuality at all temperatures tested during the first 10 days, i.e. the value did not tend to be reduced during this period. When the oysters were stored by -20 degrees C, the viability of S. typhi was apparently unaffected up to 140 days when the experiment was terminated. All these experiments were highly reproducible.

Animals↗

Tumor-associated serum proteins in rats as detected by two-dimensional gel electrophoresis.

The sera from normal and tumor-bearing rats were separated by polyacrylamide gel disc isoelectric electrophoresis and then in a perpendicular dimension by polyacrylamide gel slab electrophoresis with a gradient from 4% to 20%. When the gel was stained with Coomassie brilliant blue, 100-150 components were separately observable with normal serum. With the sera from Rhodamine sarcoma-bearing rats, three more components were detected with a high reproducibility. They were of pI (isoelectric point) 4.5 and Rm (relative mobility) 0.63, of pI4.8 and Rm 0.63 and of pI 4.9 and Rm 0.75. The 4.5/0.63 (pI/Rm) and the 4.8/0.63 components appeared soon after transplanting the tumor, whereas the 4.9/0.75 component appeared at the late stage of its growth. Upon surgical removal of the grown tumor, the former two components disappeared rapidly, while the latter component disappeared gradually. Similar results were obtained with Yoshida sarcoma, and ascites hepatoma AH66 and AH130. A 3rd polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate revealed that the 4.5/0.63 and the 4.8/0.63 components were each composed of a single polypeptide of approximately 64,000 daltons, and that the 4.9/0.75 component was composed of two different polypeptides of approximately 33,000 daltons and approximately 10,000 daltons. It was confirmed by an immunodiffusion experiment that none of these three proteins was alpha-fetoprotein.

Animals↗

[Auditory masking patterns in forward, backward, and simultaneous-masking situations (author's transl)].

To investigate the temporal course of the frequency characteristics of the response elicited in the auditory system, pure-tone masking patterns were obtained at several time positions relative to the onset or the offset of the masker. The simultaneous masking patterns obtained suggest that the shape of the response distribution remained almost unchanged during the masker, although they are less informative compared to the other conditions because of interaction effects between the masker and the probe. The forward masking data illustrate the decay course of the distribution and are believed to give more reliable informations as to the frequency selectivity of the auditory system. The backward masking data, however, fail to give any tenable suggestions. The phenomenon of the shift in the maximum-masking-frequency was observed in all conditions and its underlying mechanisms were discussed.

Adult↗