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Biomedical subjects

E Sugimoto

Publications and source records attributed to E Sugimoto.

At least 19 recordsLinked to original sources

Molecular cloning and characterization of genes encoding rat pancreatic cholecystokinin (CCK)-releasing peptide (monitor peptide) and pancreatic secretory trypsin inhibitor (PSTI).

The genes encoding a rat pancreatic cholecystokinin (CCK)-releasing peptide (monitor peptide) and its structurally related peptide, rat pancreatic secretory trypsin inhibitor (PSTI), have been isolated and sequenced. The two genes share extremely high sequence similarity in the 5' flanking regions, suggesting that these regions may be responsible for the characteristic coordinate expression of the two peptides.

Amino Acid Sequence

Monitor peptide gene expression is increased by exogenous CCK in the rat pancreas and in a rat pancreatic acinar cell line (AR4-2J).

Monitor peptide (CCK-releasing peptide) mRNA increased on the administration of CCK in rat pancreas and the AR4-2J pancreatic cell line. Subcutaneous injection of CCK into rats at 8 h intervals increased the level of monitor peptide mRNA in the pancreas. Concomitant injection of CCK antagonist CR-1409 strongly decreased it. The monitor peptide mRNA was also increased by CCK in AR4-2J cells and was decreased by the antagonist. These findings suggest that the plasma CCK induced by prolonged intake of a high protein diet may be responsible for the adaptative increase in the monitor peptide as well as exocrine proteases in the pancreas.

Amylases

An extract of Gymnema sylvestre leaves and purified gymnemic acid inhibits glucose-stimulated gastric inhibitory peptide secretion in rats.

Gastric inhibitory peptide release into the portal vein in response to duodenal infusion of D-glucose was studied in the presence of a leaf extract of Gymnema sylvestre, purified gymnemic acid and inhibitors of some putative glucose sensors and carriers in the intestinal lumen. Intraduodenal infusion of D-glucose significantly increased the portal immunoreactive gastric inhibitory peptide concentration in a dose-dependent manner. The increase in the portal immunoreactive gastric inhibitory peptide induced by glucose was significantly depressed by concomitantly infused leaf extract of Gymnema sylvestre, purified gymnemic acid and phlorizin but not by cytochalasin B. Mannoheptulose, which inhibits glycolysis, and procaine and lidocaine, which inhibit the vagal glucoreceptor in the lumen, did not affect portal immunoreactive gastric inhibitory peptide concentrations. These results suggest that a glucose receptor, which interacts with the leaf extract of Gymnema sylvestre, purified gymnemic acid and phlorizin, exists for the release of immunoreactive gastric inhibitory peptide and that the glucose receptor for gastric inhibitory peptide release is not likely to be identical with a glucose transporter or a vagal glucoreceptor in the lumen.

3-O-Methylglucose

Effects of chronic hypoxia on the whole-body insulin action in rats.

The effect of chronic hypoxia on the whole-body insulin action in rats was investigated. Rats were kept in a hypobaric hypoxia chamber maintained at a simulated altitude of 4000 m for 10 weeks. At the end of the experimental period, the mean body weight of the hypoxic rats was significantly lower than that of the control rats. The muscle weight to body ratio of the quadriceps muscle in hypoxic rats was larger than that in control rats, but those of the gastrocnemius, soleus, and extensor digitorum longus muscles did not differ between the control and hypoxic rats. On the other hand, the epididymal fat pads of hypoxic rats were markedly smaller than those of the control rats. The results of a euglycemic clamp experiment with infusions of 14 and 3.6 mU insulin.kg-1.min-1 indicated that the steady-state glucose infusion rate was not statistically different between hypoxic and control rats. It is suggested that chronic hypoxia did not influence the whole-body insulin action on glucose transport activity.

Adipose Tissue

Lipoprotein lipase enzyme expression in 3T3-L1 adipocytes is posttranscriptionally down-regulated by retinoic acid.

The effects of all-trans retinoic acid (RA) on the lipoprotein lipase (LPL) activity, synthesis and mRNA content in 3T3-L1 adipocytes were studied. When fully differentiated 3T3-L1 adipocytes were exposed to RA, dose-dependent suppression of LPL activity was observed. The loss of activity reached a maximum of 60% of the control level and appeared to be due to an effect on synthesis of the enzyme, as judged from the decreased incorporation of [35S] methionine and [35S] cysteine into immunoprecipitable LPL. The LPL mRNA level remained unchanged under the same conditions. In contrast, no significant reduction in glycerol-3-phosphate dehydrogenase activity or change in the morphological signature occurred on 24 hr exposure of 3T3-L1 adipocytes to RA. These results suggest that RA can specifically down-regulate LPL enzyme expression in adipocytes at the posttranscriptional level.

Adipose Tissue

Effect of a high-protein diet on the gene expression of a trypsin-sensitive, cholecystokinin-releasing peptide (monitor peptide) in the pancreas.

The adaptation to a high protein diet of the concentration and mRNA level of a trypsin-sensitive, cholecystokinin-releasing peptide (monitor peptide), which was proposed to be the mediator of the cholecystokinin release in response to protein intake, was investigated in the rat pancreas. Adult rats were placed on one of two isocaloric diets. One group was fed a 22% casein diet (control diet) and the other a 64% casein diet (high-protein diet) for 14 days. In order to quantify the monitor peptide separately from pancreatic secretory trypsin inhibitor (PSTI-II), which is highly similar in its amino acid and mRNA nucleotide sequences to the monitor peptide but has less cholecystokinin-releasing activity, we used specific assay methods: HPLC was used for determining the monitor peptide concentration in zymogen granules and a synthetic oligonucleotide probe for determining the mRNA of the monitor peptide in the pancreas. The concentrations in the zymogen granules and the mRNA levels in the pancreas of the two peptides increased in parallel during the adaptation to the high protein diet, indicating that these two peptides were under the same control during the adaptation. The concentration and mRNA level of the monitor peptide, which were measured after 0, 3, and 14 days, increased throughout the experiment period, as did the concentration of trypsin. This suggested that the monitor peptide and trypsin may respond to similar signals during the adaptation to a high protein diet and that this apparent coordination may facilitate the adaptation of the pancreas to the diet.

Amino Acid Sequence

Decrease in muscle glucose transporter number in chronic physical inactivity in rats.

In this study, whole body insulin action on glucose uptake and muscle glucose transporter number of rats subjected to 14 days of physical inactivity conditions was examined. Unlike other suspension and denervation models of muscle disuse, this physical inactivity model allows voluntary contractile activity with minimal stress. Minimal depression of body weight gain and significant depression of gastrocnemius muscle growth were observed compared with that of control rats after 14 days of physical inactivity. The whole body insulin sensitivity and responsiveness were determined by the euglycemic clamp technique, with 1.4, 3.6, and 14 mU insulin.kg-1.min-1 perfusion and 2-deoxy-D-[3H]glucose incorporation. The rates of glucose disposal were the same in the restrained rats as in the controls with the 1.4 and 3.6 mU insulin perfusion; however, glucose disposal significantly decreased with 14 mU insulin perfusion. 2-Deoxy-D-[3H]glucose uptake into the gastrocnemius muscle was higher in the control rats than in the physically inactive rats. Glucose transporters in the gastrocnemius and quadriceps muscles, measured by means of the D-glucose-inhibitable cytochalasin B binding assay, were significantly decreased in number in the physically inactive rats. These findings suggest that the decrease in whole body glucose uptake might in part be explained by the decreases in the total glucose transporter number in muscles.

Adrenocorticotropic Hormone

Protein factor obtained from rat adipose tissue specifically permits the proliferation of the 3T3-L1 and Ob1771 cell lines.

We have found the presence of protein factor in rat adipose tissue which permits the proliferation of 3T3-L1 and Ob1771 preadipocytes cultured in a completely defined serum-free medium containing only progression factors [epidermal growth factor (EGF) and insulin] as growth factors. This mitogenic activity of the protein factor was not detected in various other cell lines, in particular, Swiss 3T3 cells which could proliferate in response to a competent factor [platelet-derived growth factor (PDGF) or fibroblast growth factor (FGF)] in the same serum-free medium. This activity of the factor was heat- and pronase-unstable, and reductant-stable, and the apparent molecular weight of the factor was about 20,000. These results strongly suggest that the protein factor is different from PDGF or FGF and contributes to the formation of new adipocytes by specifically stimulating the proliferation of preadipocytes, acting like competent factor.

Adipose Tissue

Comparative investigation of vitamins and their analogues on terminal differentiation, from preadipocytes to adipocytes, of 3T3-L1 cells.

1. The effects of 20 kinds of vitamins or their analogues on the growth rate of preadipocytes and the terminal differentiation of preadipocytes to adipocytes was systematically compared in 3T3-L1 cells. 2. The addition of vitamin C markedly increased the growth rate of preadipocytes at over 50 microM. 3. The addition of vitamin K3 slowed down the growth rate at over 0.1 microM. 4. In water soluble vitamins and their analogues tested, the vitamin B6 group and vitamin C significantly stimulated the differentiation, and consequently increased the glycerophosphate dehydrogenase activity and triglyceride accumulation, to a concentration of over 10 microM. 5. Many fat soluble vitamins and their analogues (the vitamin A group, including beta-carotene, the vitamin D group, vitamin E and the vitamin K group) strongly inhibited the adipose conversion of 3T3-L1 cells at microM level.

Adipose Tissue

Proliferation of 3T3-L1 preadipocytes in a completely defined serum-free medium.

We have developed a completely defined serum-free medium that supports the growth of Swiss 3T3-L1 fibroblasts to nearly the same extent as Dulbecco's modified Eagle's medium supplemented with 10% fetal calf serum. With ASF301 medium [former name, RITC 80-7; Yamane et al. Exp. Cell Res. 134, 470 (1981)], most of the 3T3-L1 cells survived for at least 10 days, but did not grow. ASF301 medium contains insulin and mouse-epidermal growth factor as growth factors, which are termed "progression factors". So we examined the effects of "competent factors" on the proliferation of 3T3-L1 preadipocytes. Growth in the medium supplemented with competent factors reached a confluent monolayer in 6-7 days. Furthermore, it was confirmed that 3T3-L1 cells grown in the serum-free medium retained the properties of differentiation into adipocytes. Our serum-free medium should be a useful tool for research on the growth and differentiation of 3T3-L1 preadipocytes.

Adipose Tissue

Luminal CCK-releasing factors in the isolated vascularly perfused rat duodenojejunum.

The factors operating at the apical side of the endocrine cell releasing cholecystokinin (CCK) were investigated using the isolated vascularly perfused rat duodenojejunum. In the protease-free intestinal segment, a 30-min infusion of glucose (280 mM), oleic acid (100 mM), or triglycerides containing short- or long-chain fatty acids did not alter significantly the basal level of portal CCK-like immunoreactivity (CCK-LI), while octanoic acid (100 mM) produced a transient rise of plasma CCK-LI to approximately 250% of basal. Infusion of proteins (5% solutions of ovalbumin or casein) or of a mixture of all amino acids brought about a modest CCK secretion. In contrast, isocaloric amounts of an ovalbumin hydrolysate produced a sharp rise of portal CCK-LI to 530% of basal followed by a well-sustained plateau secretion (420% of basal) until the end of the infusion. An acid casein hydrolysate induced a slightly less pronounced CCK-LI release and was followed in decreasing order by meat, casein, and soybean peptones. Simultaneous infusion of trypsin with ovalbumin or casein hydrolysate reduced by approximately 60% the CCK release induced by peptone alone. This effect was reversed by coinfusion of soybean trypsin inhibitor (SBTI) with the trypsin-peptone mixture. Arterial infusion of tetrodotoxin (10(-6) M) or atropine (10(-5) M) had no significant effect on the trypsin-induced inhibition of peptone-mediated CCK-LI release. Administration of SBTI or camostate alone or in combination with trypsin did not alter basal CCK. Monitor peptide produced a dose-dependent transient rise of portal CCK-LI over the range from 2 to 12 micrograms.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acids

Differentiation of intestinal epithelial cell line (IEC-18) by an acid extract of rat small intestine.

A factor which may induce differentiation of intestinal epithelial cell lines in vitro was found in an acid extract of adult rat small intestine. The addition of a partially purified acetic acid extract of rat small intestine to IEC-18 cell culture dishes increased sucrase activity within 48 h. Thymidine incorporation markedly decreased within 24 h. Significant development of microvilli-like structures was observed on the acid extract-treated IEC-18 cells, compared with controls. This activity of rat acid extract was heat-stable and the apparent molecular weight of the factor was 400-800. These findings suggested that the factor may be related to the epithelial differentiation of rat small intestinal crypt cells.

Animals

Effect of ANP on circulating blood volume.

The effects of rat atrial natriuretic peptide (rANP) on blood volume (BV) were determined by the continuous measurement of BV, mean arterial pressure (MAP), and central venous pressure (CVP). Immediately after a single-bolus injection of rANP-(1-28), 1 nmol/100 g body wt, in conscious rats, BV began to decrease. Peak reduction of -0.22 +/- 0.03 ml/100 g body wt was reached 14.5 min after the injection. Thereafter, BV levels returned gradually to -0.08 +/- 0.03 ml/100 g body wt compared with the control value. In volume expansion experiment, the nephrectomized, anesthetized rats were divided into two groups: the control group, with only a saline infusion, and the ANP group, with an infusion of saline with rANP (1 nmol/100 g body wt). In the ANP group, increases in BV were not as great, and recovery was threefold faster than that of the control group. In the ANP group, the recovery time of BV to the starting control levels was 8.5 min, and the time constant of recovery was 3.6 +/- 0.3 min-1. The control group times were 25 min and 11.5 +/- 0.8 min-1, respectively. The effective vascular compliances were approximately 2.8 ml.mmHg-1.kg body wt-1 in both groups, and the capillary filtration coefficient was 0.47 ml.mmHg-1.min-1.kg body wt-1 in the ANP group and 0.33 ml.mmHg-1.min-1.kg body wt-1 in the control group. Thus the whole body capillary filtration coefficient was 1.5-fold higher in the ANP group than in the control group. This suggests that ANP may increase the permeability of capillaries.

Animals

Changes in blood volume and vascular compliance during body heating in rats.

The effects of hyperthermia on blood volume and effective vascular compliance were studied in control and heat acclimated rats (three weeks at 32 degrees C and 50% R.H.). Experiments were performed on conscious rats whose abdominal aorta and both jugular veins were cannulated. Continuous changes in blood volume (BV) were monitored by measuring 51Cr tagged erythrocyte dilution, using an arterio-venous extracorporeal shunt passing through a gamma counter. Total vascular compliance was calculated from the relation between changes in BV and central venous pressure during 10 min of infusion of saline at a rate of 1.6% body wt/10 min. Hyperthermia induced a significant blood volume expansion. This expansion was more pronounced in non-acclimated rats. Effective vascular compliance was similar in the normothermic, both non-acclimated and acclimated rats. However, while hyperthermia did not affect the vascular compliance of non-acclimated rats, it was decreased significantly in the acclimated hyperthermic rats. The data suggest that changes in vascular compliance play a role when rapid blood volume changes take place, especially in acclimated hyperthermic animals. The relations between changes in vascular compliance and heat induced redistribution of cardiac output are discussed.

Acclimatization

Analysis of salt and water intake by continuous determination of blood volume and plasma sodium concentration.

Seven rats were exposed to a hot environment (36 degrees C) for about 5 h to induce thermal dehydration. They were allowed to recover from their fluid loss; both tap water and 1.8% NaCl solution were provided simultaneously as drinking fluids. In the recovery stage, these animals initially consumed low NaCl solution (about 41 mEq/l) for the first 90 min and then shifted to drinking slightly hypotonic NaCl solution (about 122 mEq/l) on the average. To analyze the mechanism driving the above shift, changes in blood volume (BV), plasma Na concentration ([Na]), and the amount of water and 1.8% NaCl solution consumed were measured continuously. After 4 h of recovery, BV increased by +1.1 ml/100 g body wt. and reached 5.7 ml/100 g body wt. (set as 100%) while plasma [Na] reduced by -5.5 mEq/l or to 141.5 mEq/l (mean values of 7 rats). The shift of [Na] of drinking water was observed when plasma [Na] reduced to -3.4 mEq/l or 62.3% of the 4th-hour level, while BV recovery remained only +0.5 ml or 46.3%. A response to urinary output started later when plasma [Na] reached 142.6 mEq/l or 83.1%, while BV recovery remained +5.4 ml or 76%. These results indicate that the regulation of plasma [Na] takes precedence over the regulation of BV. This conclusion is in agreement with the view that the blood osmolality change directly modifies the cell volume whereas circulatory function under decreased blood volume can be regulated by change of the vascular compliance.

Animals

[Radiologic detection of metastatic bone tumors].

Radiologic strategy for early detection of bone metastasis is discussed. Incidence, pathway, and radiologic manifestation of bone metastasis are briefly discussed. Bone scintigraphy is a mainstay for early detection of bone metastases, although its role as a part of staging procedure is debatable. Plain radiograph and CT scan are to be used to increase specificity of scintigraphic findings. For detection of spinal metastases CT scan is particularly useful and should be used whenever spinal metastases are suspected. MRI has become an important modality to see an extraosseous extension and marrow invasion of metastatic bone tumor.

Bone Neoplasms

Lithium ion reversibly inhibits inducer-stimulated adipose conversion of 3T3-L1 cells.

Adipose conversion of 3T3-L1 cells by inducers (dexamethasone, 1-methyl-3-isobutylxanthine and insulin) was inhibited by LiCl at concentrations from 2 to 20 mM. The effect of LiCl was reversible and the inhibited cells were converted to adipocytes when stimulated after the removal of LiCl. Inhibition by LiCl of adipose conversion was accompanied with a blockage of the enhanced [3H]thymidine incorporation and cellular proliferation that occurred before the adipocyte phenotype was expressed. Of the cations tested, only Li+ had these effects.

1-Methyl-3-isobutylxanthine

Iodine-123 IMP SPECT before and after bypass surgery in a patient with occlusion of left anterior and middle cerebral arteries with basal abnormal telangiectasis (unilateral Moyamoya disease).

A case of left anterior and middle cerebral arterial occlusion with angiographic features similar to Moyamoya disease was reported. IMP SPECT of the patient revealed the success of bypass surgery clearly. The patient complained of transient right hemiparesis with aphasia 4 times. The cerebral arteriography disclosed occlusions of left anterior and middle cerebral arteries at their proximal portions. Right internal carotid and its branches were normal. I-123 IMP SPECT study showed hypoperfusion in left temporal lobe, basal ganglia with incomplete reperfusion on the delayed (4 hours after injection) SPECT images. After the superficial temporal-middle cerebral artery anastomosis, I-123 IMP SPECT showed improvement of the brain blood flow. I-123 IMP SPECT was very useful in detecting the ischemic areas and evaluating the revascularizing surgery in this case.

Adult