Impaired insulin secretion from pancreatic beta-cells in response to glucose is an important feature in the pathology of non-insulin-dependent diabetes mellitus (NIDDM). In the course of screening for useful insulin secretagogues, we have isolated and identified YHB-2017 (Genistein) as a insulin secretion potentiator from fermentation broths of our in-house microbial library. The insulinotropic activity of YHB-2017 in isolated rat pancreatic islets was exerted only at high concentration of glucose (8.3-16 mM) but not at low concentration of glucose (3.3-5.5 mM). Also, in perifusion study with isolated rat pancreatic islets, YHB-2017 stimulated insulin secretion in a time-dependent manner when YHB-2017 was added to KRB buffer containing 16 mM glucose. In the presence of $200\;{\mu}M$ diazoxide and 35 mM KCI, which stimulates maximum $Ca^{2+}$ influx independently of KATP channel, YHB-2017 enhanced KATP channel-independent insulin secretion at high concentration glucose (16 mM). To elucidate the mechanisms of the glucose-dependent potentiation effect of YHB-2017, pharmacologic inhibitors for protein kinase A, protein kinase C and calcium/calmodulin kinase II were pre-treated and then the potentiation effect of YHB-2017 on insulin secretion was investigated. Pre-treatment of H89 as a PKA inhibitor had a significant inhibitory effect on YHB-2017-induced potentiation effect. Furthermore, western immunoblotting analyses revealed that YHB-2017 increased phosphorylation of PKA substrates and cAMP response element-binding protein (CREB) under high concentration of glucose. These results demonstrated that the insulinotropic effect of YHB-2017 is mediated through PKA signal pathway and activated amplifying $K_{ATP}$ channel-independent insulin secretion pathway.
Journal of the Korean Society of Food Science and Nutrition
/
v.31
no.2
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pp.295-305
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2002
This paper, in which whose subjects were 43 cerebrovascular accident patients analysed the effects of flood habits and attitudes on the nutrient intake. In respect to energy intake, the subjects took 106% of RDA. The protein intake was on the average of 119.1 g, which was 187% of RDA. The fat intake by the subjects on the whole was 60.5 g. The fiber intake of the subjects was 9.6 g. Those who like sweets took in significantly less energy and carbohydrate and more fat than those who didn't like sweets. Those who liked salty flood took in 7890 mg of sodium while those who didn't like salty food took in 5579 mg of sodium. The former took in significantly more sodium than the latter (p < 0.05). The examination of the amount of nutrient intake in terms of meal pattern, showed that those who had two meals a day were significantly higher in the level of weight and BMI was significantly higher (p<0.05) and the level of energy, protein, calcium, iron, vitamin A, vitamin C and cholesterol was significantly higher. Those who thought they had heavy meals took significantly more energy, protein, calcium iron, vitamin A vitamin B$_1$and vitamin C than those who thought they had light meals. Rapid eaters took more nutrients than slow eaters. The multiple regression analysis has shown that the effect of the independent variables on the energy intake are in the order of eating speed, eating volume and eating frequency. They can explain 24.6% of the energy intake. As a result, the faster is eating speed, the heavier is eating volume, and the lower is eating frequency, the higher is the energy intake (p <0.01).
Nitric oxide(NO) has been reported to be one of the mediators relating to bone remodelling. Nitric oxide is synthesized from L-arguinine by nitric oxide synthetase(NOS), which is largely divided Into two groups. One group which is composed of $NOS_1\;and\;NOS_3$, is dependent of calcium or calmodulin. The other consisted of $NOS_2$, which is independent of calcium or calmodulin. NOS is thought to be a possible intermediate affecting in the course of tooth movement. This study was designed to evaluate the expression of nitrous oxide synthetase(NOS) in periodontal tissue during the experimental movement of rat incisors, by LSAB(labelled streptavidine biotin) immunohistochemical staining for $NOS_2\;and\;NOS_3$. Twenty seven Sprague-Dawley rats were divided into a control group(3 rats), and 6 experimental groups(24 rats), to which 75g of force was applied, with helical springs across the maxillary incisors. Rats of experimental groups were sacrificed at 12 hours, 1, 4, 7, 14 and 28 days after force application, respectively. After that, the tissues of the control group and experimental groups were studied immunohistochemically. The results were as follows: 1. In control group, the expression of $NOS_3$ was rare in gingiva, dentin, periodontal ligament and alveolar bone, and was mild in the capillaries of pulp and intermaxillary suture. And the expression of $NOS_2$ showed similar pattern to that of $NOS_3$. 2. There were no differences in the expression of $NOS_2\;or\;NOS_3$ in dentin, gingiva, cementum, cementoblast and odontoblast, between control and experimental groups, regardless of the duration of the force application. 3. The expression of $NOS_3$ began to increase at 4 days and showed to the highest degree at 7 days after force application, in the apical region of pressure side of periodontal ligament in experimental groups. 4. The expression of $NOS_3$ in alveolar bone was rare until 7 days, after which it increased to mild degree at 14 days through 28 days in experimental group. But there was no difference between pressure and tension side of periodontal ligament. 5. The expression of $NOS_2$ in periodontal ligament was mild from 7 days after force application, regardless of the side of periodontium, which was generally more evident than that of $NOS_3$. 6. The expression of $NOS_2$ in alveolar bone increased to mild degree at 14 days after force application, and it was evident in osteoblasts, osteoclasts and osteocytes. And the expression of $NOS_2$ was little more stronger in the tension side than that of pressure side of alveolar bone.
In order to evaluate soil factors affecting the growth of Italian poplar, 23 areas planted with Italian poplar were surveyed. These 23 areas were classified into 3 categories, river-side, fallow-land and hill-side. The growth performance and soil factors for each area were investigated. The growth of Italian poplar at river-side was shown to be superior to that of fallow-land and fill-side. The rates of growth for fallow-land and hill-side are decreased by 8% and 21% compared to those of river-side, respectively. This suggests that plantation of Italian poplar at hill-side would not be profitable. Soil conditions of high productive area appeared liquid phase 20%, porosity 45%, water holding capacity 35 - 40%, soil hardness $1kg/cm^3$. pH 6 and rich in organic matter and total nitrogen. The results of factor analysis for soil factors affecting to Italian poplar growth that showed eigenvalue over 1 and communality value over 70% explained factor 1 : liquid phase, porosity and water holding capacity, factor 2 : pH and calcium, and factor 3 : soil hardness. This suggests that physical characteristics of soil is more important than chemical characteristics for Italian poplar growth. Multiregerssion analysis was conducted between diameter growth and soil hardness, liquid phase and calcium. The t-values for each independent variables showed significance at 1 - 10% level, but water holding capacity and pH are not significant. It is supposed that sites suitable to Italian poplar were alluvial plain of sandy loam or part of banking soil, well-ventilating soil, lower soil hardness, apposite soil moisture absorbing with about 100cm of ground water level, plentiful organic matters and total nitrogen and little acidity soil.
Journal of the Korean Society of Food Science and Nutrition
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v.24
no.1
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pp.30-40
/
1995
The relationship between dietary intake and vitamin/mineral supplement usage was examined in 706 adolescent girls who were high school students. 43.8% of subjects used vitamin/mineral supplements during one year. The higher the family income and parents education level, the higher percentage of vitamin/mineral supplement usage was. But there was no significant difference between grades and scores. And vitamin/mineral supplement usage was higher int he thin and obese groups than the average weight groups. Nutrition knowledge and food habit did not affect vitamin/mineral supplement usage. Calorie intakes of vitamin/mineral supplement users and nonusers were similar. However, independent of the supplements, the diets of supplement nonusers contained significantly more dietary protein, vitamin A, vitamin $B_2$, niacin, vitamin C, and calcium than the diets of the users. A considerable portion of both the users and nonusers had dietary intakes of less than 2/3 of the Recommended Dietary Allowances for vitamin $B_1$, vitamin $B_2$, niacin, calcium, and iron. Vitamin/mineral supplement nonusers generally consumed a more vitamin, mineral from diet. Reasons for taking supplements were to take energy, advice and illness.
This study was conducted to evaluate the relative bioavailability (RBV) of 25-hydroxycholecalciferol (25-OH-$D_3$) to cholecalciferol (vitamin $D_3$) in 1- to 21-d-old broiler chickens fed with calcium (Ca)- and phosphorus (P)-deficient diets. On the day of hatch, 450 female Ross 308 broiler chickens were assigned to nine treatments, with five replicates of ten birds each. The basal diet contained 0.50% Ca and 0.25% non-phytate phosphorus (NPP) and was not supplemented with vitamin D. Vitamin $D_3$ was fed at 0, 2.5, 5.0, 10.0, and $20.0{\mu}g/kg$, and 25-OH-$D_3$ was fed at 1.25, 2.5, 5.0, and $10.0{\mu}g/kg$. The RBV of 25-OH-$D_3$ was determined using vitamin $D_3$ as the standard source by the slope ratio method. Vitamin $D_3$ and 25-OH-$D_3$ intake was used as the independent variable for regression analysis. The linear relationships between the level of vitamin $D_3$ or 25-OH-$D_3$ and body weight gain (BWG) and the weight, length, ash weight, and the percentage of ash, Ca, and P in femur, tibia, and metatarsus of broiler chickens were observed. Using BWG as the criterion, the RBV value of 25-OH-$D_3$ to vitamin $D_3$ was 1.85. Using the mineralization of the femur, tibia, and metatarsus as criteria, the RBV of 25-OH-$D_3$ to vitamin $D_3$ ranged from 1.82 to 2.45, 1.86 to 2.52, and 1.65 to 2.05, respectively. These data indicate that 25-OH-$D_3$ is approximately 2.03 times as active as vitamin $D_3$ in promoting growth performance and bone mineralization in broiler chicken diets.
Kim, Nam-Su;Kim, Jin-Ho;Kim, Hwa-Seong;Kim, Hui-Seon;Lee, Seong-Su;Todd, Andrew C.;Lee, Byeong-Guk
Journal of Korean Society of Occupational and Environmental Hygiene
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v.16
no.4
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pp.324-333
/
2006
This study was designed to investigate the effect of bone demineralization and tibia lead on blood lead in retired lead workers. Two hundred thirty five(126 females and 109 males) retired lead workers who worked in 4 different lead factories and 101 non-occupationally lead exposed subjects(51 females and 51 males) were recruited from March 2004 to October 2004. Bone mineral density(BMD) was measured at left calcaneous bone area by broadband ultrasound attenuation(BUA) method with QUS-2(Metra Biosystems Inc, USA). The BUA value transformed into T-score by WHO standard conversion criteria. Tibia bone lead was measured for skeletal bone lead with K-xray fluorescence(K-XRF) and blood lead was analyzed with flameless atomic spectrophotometer. Hemoglobin, hematocrit, serum calcium and iron were also analyzed. In addition, information for smoking and drinking status and basic personal data such as age, gender and lead exposure were also collected using questionnaire inquiry. Blood lead was correlated with tibia lead (r=0.664) and these two variables were negatively correlated with BMD in bivariate analysis. BMD showed significant main effect on the change of blood lead independent to tibia lead without any effect modification of age or gender; the one T-score unit decrease of mineral bone density made $0.43{\mu}g/dl$ increase of blood lead. On the other hand, tibia lead showed effect modification with gender on blood lead; the slope of tibia lead on blood lead in male was steeper than in female and crossed at around zero of tibia lead. In the multiple regression analysis of blood lead and tibia lead on BMD after adjustment of related covariates, only blood lead showed statistically significant effect on BMD. This study confirmed that BMD and blood lead were significantly associated. To verify the causal association of BMD on blood lead and vice versa, further longitudinal studies are needed.
Phenolic compounds affect intracellular free $Ca^{2+}$ concentration ($[Ca^{2+}]_i$) signaling. The study examined whether the simple phenolic compound octyl gallate affects ATP-induced $Ca^{2+}$ signaling in PC12 cells using fura-2-based digital $Ca^{2+}$ imaging and whole-cell patch clamping. Treatment with ATP ($100\;{\mu}M$) for 90 s induced increases in $[Ca^{2+}]_i$ in PC12 cells. Pretreatment with octyl gallate (100 nM to $20\;{\mu}M$) for 10 min inhibited the ATP-induced $[Ca^{2+}]_i$ response in a concentration-dependent manner ($IC_{50}=2.84\;{\mu}M$). Treatment with octyl gallate ($3\;{\mu}M$) for 10 min significantly inhibited the ATP-induced response following the removal of extracellular $Ca^{2+}$ with nominally $Ca^{2+}$-free HEPES HBSS or depletion of intracellular $Ca^{2+}$ stores with thapsigargin ($1\;{\mu}M$). Treatment for 10 min with the L-type $Ca^{2+}$ channel antagonist nimodipine ($1\;{\mu}M$) significantly inhibited the ATP-induced $[Ca^{2+}]_i$ increase, and treatment with octyl gallate further inhibited the ATP-induced response. Treatment with octyl gallate significantly inhibited the $[Ca^{2+}]_i$ increase induced by 50 mM KCI. Pretreatment with protein kinase C inhibitors staurosporin (100 nM) and GF109203X (300 nM), or the tyrosine kinase inhibitor genistein ($50\;{\mu}M$) did not significantly affect the inhibitory effects of octyl gallate on the ATP-induced response. Treatment with octyl gallate markedly inhibited the ATP-induced currents. Therefore, we conclude that octyl gallate inhibits ATP-induced $[Ca^{2+}]_i$ increase in PC12 cells by inhibiting both non-selective P2X receptor-mediated influx of $Ca^{2+}$ from extracellular space and P2Y receptor-induced release of $Ca^{2+}$ from intracellular stores in protein kinase-independent manner. In addition, octyl gallate inhibits the ATP-induced $Ca^{2+}$ responses by inhibiting the secondary activation of voltage-gated $Ca^{2+}$ channels.
A gene encoding a putative phospholipase D was isolated from Bacillus licheniformis and cloned into pGEM-T easy vector. The gene was expressed in E. coli BL21 (DE3) using a pET-21(a) vector containing His6 tag. Affinity purification of the recombinant phospholipase D with nickel-nitrilotriacetic acid (Ni-NTA) resin resulted major one-band by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis. The purified enzyme showed a molecular weight of 44 kDa. The optimum activity of enzyme was around pH 7.0 and the enzyme was also the most stable around this condition. The optimum temperature was about $40-45^{\circ}C$ and the enzyme still showed considerable activities at wide range of temperature. Among various detergents, Triton X-100 significantly increased the enzyme activity, resulting in 181% activity of control at 0.6 mM of the detergent. Calcium ion did not significantly affect the enzyme activity, suggesting that the enzyme might be classified into $Ca^{2+}$-independent PLD.
Park, Su-Jung;Choi, Won-Woo;Kwon, Oh-Sang;Chung, Jin-Ho;Eun, Hee-Chul;Earm, Young-E;Kim, Sung-Joon
The Korean Journal of Physiology and Pharmacology
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v.12
no.4
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pp.177-183
/
2008
The layers of keratinocytes form an acid mantle on the surface of the skin. Herein, we investigated the effects of acidic pH on the membrane current and $[Ca^{2+}]_c$ of human primary keratinocytes from foreskins and human keratinocyte cell line (HaCaT). Acidic extracellular pH ($pH_e{\leq}5.5$) activated outwardly rectifying $Cl^-$ current ($I_{Cl,pH}$) with slow kinetics of voltage-dependent activation. $I_{Cl,pH}$ was potently inhibited by an anion channel blocker 4,4'-diisothiocyanostilbene-2,2'-disulphonic acid (DIDS, 73.5% inhibition at 1${\mu}$M). $I_{Cl,pH}$ became more sensitive to $pH_e$ by raising temperature from $24^{circ}C$ to $37^{circ}C$. HaCaT cells also expressed $Ca^{2+}$-activated $Cl^-$ current ($I_{Cl,Ca}$), and the amplitude of $I_{Cl,Ca}$ was increased by relatively weak acidic $pH_e$ (7.0 and 6.8). Interestingly, the acidic $pH_e$ (5.0) also induced a sharp increase in the intracellular [$Ca^{2+}$] (${\triangle}[Ca^{2+}]_{acid}$) of HaCaT cells. The ${\triangle}[Ca^{2+}]_{acid}$ was independent of extracellular $Ca^{2+}$, and was abolished by the pretreatment with PLC inhibitor, U73122. In primary human keratinocytes, 5 out of 28 tested cells showed ${\triangle}[Ca^{2+}]_{acid}$. In summary, we found $I_{Cl,pH}$ and ${\triangle}[Ca^{2+}]_{acid}$ in human keratinocytes, and these ionic signals might have implication in pathophysiological responses and differentiation of epidermal keratinocytes.
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