• Applied Microscopy
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• v.24 no.2
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• pp.78-92
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• 1994

Lysozyme has been reported to be present in the secretory granules of the Paneth cell, and lysozyme immunoreactivity has been detected by immunogold method in Paneth cells of the intestine of human, mouse and rat. The present study was aimed at clarifying the intracellular distribution and changes of the lysozyme immunoreactivity in rabbit Paneth cell after common bile duct ligation of rabbit, using the electron microscope immunogold technique. Healthy adult rabbits weighing about 2kg body weight were divided into normal and bile duct ligated groups. Common bile duct ligation was performed aseptically under ether anesthesia. Experimental animals were sacrificed on the 1st, the 3rd, the 5th, the 7th and the 14th day after the operation. Mucosal specimens from the intestinal gland of ileum were fixed in 2.5% glutaraldehyde-1.5% paraformaldehyde, followed by 1% osmium tetroxide, embedded in araldite mixture, cut with LKB-V ultratome. Ultrathin sections were placed on parlodion coated nickel grids (200mesh). The section-bearing grids were floated upside down on the added substance in a moist chamber at room temperature except for the primary antibody step, which was at $4^{\circ}C$. Sections were etched with a saturated solution of sodium m-periodate for 60min. After etching, sections were pretreated with 0.02M tris buffered saline (TBS), pH 8.4, with 1% bovine serum albumin (BSA, Sigma) for 60min, then treated polyclonal rabbit anti-human lysozyme (Dakipatts) diluted 1 : 50 in TBS with 0.1% BSA for 20hr. Subsequently, grids were incubated 60min in biotinylated goat anti rabbit IgG (Amersham) diluted 1 : 100 in TBS with 0.1% BSA. After this, sections were incubated 60min on streptavidin gold G10 (Amersham) diluted 1 : 50 in TBS with 0.1% BSA. After each step, the grids were briefly rinsed with TBS with 0.1% BSA. After the strepavidin gold step, the sections were jet washed with distilled water. Counterstain of the sections performed by uranyl acetate and lead citrate, and observed with JEM 100 CX II electron microscope. Observed results were as follow; 1. Secretory granules of mouse Paneth cells have a lysozyme immunoreactivity and also eosinophil leucocyte of rabbit applied for the positive-control stain, are well labeld with gold particles. 2. Normal rabbit Paneth cells have a lysozyme immunoreactivity restricted on the secretory granules. 3. Amount lysosomes containing myelin figures in the Paneth cells were significantly increased from 5th day after the common bile duct ligation. 4. Immunoreactivity of Paneth cell secretory granules were more activated on the 3rd day after the common bile duct ligation as compared with those of the normal animal. But the lysozyme immunoreactivity were decreased from the 5th day after the common bile duct ligation. 5. Considering the above finding, lysozyme contained Paneth cell are affected following of common bile duct ligation, whereas lysosomes containing myelin-figure do not exhibit any immunoreactive relationship with those of secretory granules.

• The Korean Journal of Nuclear Medicine
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• v.32 no.1
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• pp.50-60
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• 1998

I-123 labelled fatty acids are suitable for investigation of regional myocardial metabolism, so they are on the clinical trial. However, the precise properties of these materials are not characterized yet. We have synthesized phenylpentadecanoic acid and labeled this compound with I-123. The purpose of this study was to examine the stability, biodistribution, metabolism and SPECT imaging of [I-123]15-(p-iodophenyl)pentadecanoic acid(I-123-IPPA) that we made. The stability test of I-123-IPPA in serum of rat, mouse and human showed no free I-123 after 1 hour. In biodistribution study in mice for various time intervals after injection(5, 10, 15, 30, 60 minutes), uptake in myocardium was 14.5%ID/g(5 min), and 1.9%ID/heart(5 min), while uptake in muscles was 2.6%ID/g(5 min). Myocardium to blood ratio and myocardium to lung ratio increased for 5 min after injection and then decreased rapidly. Chromatographic data of rat blood and urine showed that little PPA was found in blood and urine at 15-20 min after injection. The myocardial I-123-IPPA SPECT images of a dog with myocardial infarction showed defects similar to those of Tc-99m-MIBI and F-18-FDG. These data suggest that I-123-IPPA is quite stable in vitro and shows favorable biodistribution in mice. SPECT imaging with I-123-IPPA demonstrated infarct zone as photon defect in dog model of myocardial infarction. I-123-IPPA may be used for the evaluation of fatty acid metabolism in clinical trials in Korea.

• Journal of Sasang Constitutional Medicine
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• v.8 no.1
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• pp.295-317
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• 1996

It is researched to elucidate the effects of Taeumjowuitang(TE,太陰調胃湯), Sibimikwanjungtang(SE, 十二味寬中湯) and Yangkeogsanwhatang(SY,凉膈散火湯) on the obesity induced by gold thioglucose and the differentiation and growth of preadipocyte 3T3-L1 in the mouse. The result were as follows: 1. TE,SE and SY extracts improved the blood level of transaminase in the obese mouse induced by gold thioglucose. 2. TE,SE and SY extracts inhibited the increase of liver fat and body fat in the obese mouse induced by gold thioglucose. 3. TE,SE and SY extracts inhibited the increase of body weight in the obese mouse induced by gold thioglucose. 4. TE,SE and SY extracts inhibited the growth of undifferentiate preadipocyte 3T3-L1. 5. TE,SE and SY extracts showed inhibitory effect on the differentiation of preadipocyte 3T3-L1. The above results suggest that the TE,SE and SY extracts may be used on the obesity induced by the overgrowth and differentiation of adipocyte, and the accumulation of fat in liver and body.

• Journal of the Korean Society of Food Science and Nutrition
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• v.12 no.3
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• pp.273-283
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• 1983

In order to investigate the effect of the combined dietary animal and vegetable proteins on growing female and male rats and mice. 25 female and 25 male rats of 4 weeks old weighing approximately 43-65g and 47-60g, respectively, were subjected to feeding trials for 4 weeks and then subsequently to metabolic trials for 10 days. In case of mice, 25 female and 25 male mice of 4 weeks old weighing approximately 12-17g and 12-16g, respectively, were equally treated. The dietary energy level was set as 3600 kcal ME/kg, and protein and fat provided respectively 12 and 5% of the dietary energy. The rest 83% of the energy level was supplied with appropriate amounts of starch and glucose. The following results were obtained. The body weight gain of female and male rats were increased as the combined dietary animal protein level increased. Whereas that of male mice was the highest for the diet E(Ap 10+Vp 90). Food efficiencies both of female and male rats were improved as combined dietary of animal protein increased. Male rats were superior to that of female. In the mice, the food efficiency value of male mice was superior to that of female. And the values of the male showed the same fashion as that in the growth rate, although it was not for the female. Protein efficiencies, both of female and male rats, were improved as combined dietary of animal protein level increased. That of male mice was in accord with the body weight gain and food efficiencies. But that of female mice was not accordant. Nitrogen retention of the experimental diets in the same protein level was in proportion to nitrogen intake. That of male mice was improved as animal protein level increased, but, that of female mice was not. Body protein utilizability was found to be superior in mice to rats. Blood serum protein level was not found to be affected by the experimental diets in the same protein level. The best combination ratio of animal and vegetable protein seems to be variable depending on the sex and species of animals, and the best diets for female and male rats and female and male mice were found to be experimental diet A(Ap 10+Vp 90) or diet B(Ap 75+Vp 25), diet C(Ap 50+Vp 50), and diet E(Ap 10+Vp 90), respectively. From the above-mentioned results that there was no difference in growth rate of body weight gain from the weanling period to puberty, and that the protein requirements of rats and mice were different from each other.

• Journal of the Korean Society of Food Science and Nutrition
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• v.40 no.5
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• pp.682-688
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• 2011

The purpose of this study was to measure the blood glucose level and glycemic index (GI) in response to persimmon (Diospyros kaki Thunb) syrup extracted from persimmon and extract of persimmon by-products. Major component analyses of persimmon syrup I (PSI, 95:5 mixture of purified persimmon syrup and non-purified persimmon syrup) and persimmon syrup II (PSII, 50:50 mixture ratio of purified persimmon syrup and non-purified persimmon syrup) were $0.3{\pm}0.1$ and $0.6{\pm}0.2$ mg/g for total polyphenolic compounds and $70.6{\pm}0.6$ and $66.6{\pm}1.6%$ for total carbohydrates, respectively. Blood glucose responses of PSI and PSII were determined using both normal ICR mice and streptozotocin (STZ)-induced diabetic male Sprague-Dawley (SD) rats. Further, oral glucose tolerance test (OGTT) was performed on diabetic rats to assess the effects of the experimental diets. Blood glucose response and OGTT showed that blood glucose levels were significantly lower in mice and diabetic rats fed PSI and PSII compared to those fed diets of sugar, maple syrup, or honey. The GIs of healthy volunteers in response to PSI and PSII were calculated to be 51.9 and 35.7, respectively. On the contrary, the GIs of healthy volunteers fed diets including sugar, maple syrup, or honey were 52.6, 20.0, and 93.0, respectively. These results suggest that persimmon syrup can be used for both the treatment of diabetics and healthy people due to its beneficial effects on blood glucose level.

• Journal of Food Hygiene and Safety
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• v.37 no.3
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• pp.189-197
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• 2022

Asthma is a chronic inflammatory disease characterized by recurring symptoms, airflow obstruction, and bronchial hyper-responsiveness. The onset of asthma for most patients begins early in life, and current asthma treatment with anti-inflammatory agents can have adverse effects, eventually leading to impaired quality of life. In the pathogenesis of asthma, macrophages and basophils play a vital role during progression. Macrophages not only induce inflammation by secreting inflammatory cytokines but also promote DNA damage and mucus production through nitric oxide (NO) production. Basophils enhance eosinophil recruitment and aggravate asthma through the FcεRIα receptor with high affinity for histamine and IgE. Therefore, in this study, we investigated whether the activation of macrophages and basophils is suppressed by the individual extracts of 28 natural products. RAW 264.7 cells (mouse macrophages) were treated with the natural products in LPS, and 4 natural product extracts resulted in decreased NO production. In β-hexosaminidase assay using RBL-2H3 cells (rat basophils), 19 natural product extracts decreased β-hexosaminidase production. In NO production and β-hexosaminidase assay using macrophages and basophils, 3 natural product extracts (Plantago asiatica, Centella asiatica, and Perilla frutescens var. japonica) significantly inhibited NO production and β-hexosaminidase release. Overall, we examined the inhibitory effects of 28 natural product extracts on macrophage and basophil activity, and the findings demonstrated the potential of natural product extracts for treating asthma and macrophage- and basophil-related diseases.

• Korean Journal of Food Science and Technology
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• v.35 no.5
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• pp.980-987
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• 2003

In this report, we investigated the detoxication effects of Saururus chinenis, Geranium nepalense, Lonicera japonica, Cassia obtusifolia, Glycyrrhiza uralensis, or their mixtures by employing acute toxicity tests for nicotine and dioxin. When fatal doses $(LD_{100}\;=\;42\;mg/kg)$ of nicotine were injected into the abdominal cavities of ICR mice, those treated with OHEM showed delayed paralysis, half the duration of hyperactivity, and a 73 % survival rate. The results revealed the strong detoxicating effects of the mixtures. We also measured the amount of the degradation product of nicotine and cotinine in humans. Consumption of OHEM promoted (he more specific) the metabolic pathways of nicotine, increasing continine excretion by 1.5 times. As a result the amount of cotinine in urine was reduced to less than 5% after treatment with OHEM. In order to test the toxicity of dioxin, we used TcnN(SD)BR rats exposed to TCDD. While TCDD treatment reduced the blood levels of hemoglobin and platelet, OHEM consumption relieved these effects and, furthermore, helped to recover the number of platelet to the normal level (p<0.05). Moreover, neutrophils (%) and monocytes (%), which were reduced by the injection of TCDD, recovered to normal levels upon treatment with OHEM. The amount albumin reduced by TCDD (p<0.05) normalized, while the activities of GOT and GTP increased by TCDD were reduced. Increases in total cholesterol and neutral fatty acids induced by TCDD were also reduced by OHEM injection (p<0.05). In the kidney, TCDD-induced rises in creatinine were suppressed by OHEM treatment, while decreases in iron levels from TCDD were raised to normal. The treatment of TCDD had more toxic effects in the blood and pancreas than on the liver, kidney and heart. On the other hand, the detoxication of OHEM had significant effects on the liver and pancreas. The normalization by OHEM of various clinical abnormalities induced by TCDD demonstrates the detoxicating effect of OHEM that ameliorates systemic metabolism not properly functioning.

• Applied Microscopy
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• v.39 no.2
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• pp.167-173
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• 2009

Capillaria hepatica is a parasitic nematode which causes hepatic capillariasis in rodents and other mammals, including man. Rat species of the genus Rattus are main primary host and rates of genus Rattus of up to 100% have been reported. Infection to reservoir and other mammalian hosts occur incidentally due to ingestion of water or food contaminated with C. hepatica embryonated eggs. The worms mature exclusively inside the liver, but they die and disassemble soon after egg spawning in rats. Dead worms and their eggs cause immune response of focal necrosis and inflammation within the liver. C. hepatica adult with a thin and long body is similar to capillary. The members of Order Trichurida are characterized by having a stichosome and the bacillary bands in front of the body. As already mentioned, the adult C. hepatica residesin the liver, where it deposits groups of eggs, and finally die in the encapsulated tissue of the liver. They produce eggs that elicit a marked granulomatous reaction that eventually destroy the worms. And the adult worms were mixed with eggs. So the complete isolation of the worm and observation of intact ultrastructure is very difficult. In this study, integument structure of C. hepatica isolated from the liver of mouse at 7 weeks after inoculation of embryonated eggs were observed with scanning and transmission electron microscopy. As a results, body length of isolated C. hepatica was about 99 mm. Cuticle, bacillary band and bacillary pore were obtained in the integument of worm. Bacillary pore across cuticular surface of the worm were observed. According to the existence of cap material, external forms of bacillary pore can be divided into three types such as flat, ingression, and ingression with the cap material type. The complete isolation of the worm and observation of ultrastructure of integument will provide the fundamental data which is important in the nematode research including C. hepatica.

• Tuberculosis and Respiratory Diseases
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• v.53 no.6
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• pp.619-634
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• 2002

Background : Many inflammatory mediators and collagenases are involved in the pathogenesis of acute lung injury (ALI) and acute respiratory distress syndrome (ARDS). The increase of matrix metalloproteinase-9 (MMP-9, gelatinase-B) produced mainly by inflammatory cells was reported in many ALI models and connective tissue cells. In this study, the expression of MMP-9 in ventilator-induced lung injury (VILI) model and the effects of matrix metalloproteinase inhibitor (MMPI) on VILI were investigated. Methods : Eighteen Sprague-Dawley rats were divided into three groups: low tidal Volume (LVT, 7mL/Kg tidal volume, 3 $cmH_2O$ PEEP, 40/min), high tidal volume (HVT, 30mL/Kg tidal volume, no PEEP, 40/min) and high tidal volume with MMPI (HVT+MMPI) groups. Mechanical ventilation was performed in room air for 2 hours. The 20 mg/Kg of CMT-3 (chemically modified tetracycline-3, 6-demethyl 6-deoxy 4-dedimethylamino tetracycline) was gavaged as MMPI from three days before mechanical ventilation. The degree of lung injury was measured with wet-to-dry weight ratio and acute lung injury score. Expression of MMP-9 was studied by immunohistochemical stain with a mouse monoclonal anti-rat MMP-9 $IgG_1$. Results : In the LVT, HVT and HVT+MMPI groups, the wet-to-dry weight ratio was $4.70{\pm}0.14$, $6.82{\pm}1.28$ and $4.92{\pm}0.98$, respectively. In the HVT group, the ratio was significantly higher than other groups (p<0.05). Acute lung injury score measured by five-point scale was $3.25{\pm}1.17$, $12.83{\pm}1.17$ and $4.67{\pm}0.52$, respectively. The HVT group was significantly damaged by VILI and MMPI protects injuries by mechanical ventilation (p<0.05). Expression of MMP-9 measured by four-point scale was $3.33{\pm}2.07$, $12.17{\pm}2.79$ and $3.60{\pm}1.95$, respectively, which were significantly higher in the HVT group (p<0.05). Conclusion : VILI increases significantly the expression of MMP-9 and MMPI prevents lung injury induced by mechanical ventilation through the inhibition of MMP-9.

• Journal of Life Science
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• v.24 no.4
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• pp.343-351
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• 2014

Phosphoinositide 3-kinase (PI3K) plays a central role in cell signaling and leads to cell proliferation, survival, motility, exocytosis, and cytoskeletal rearrangements, as well as specialized cell responses, superoxide production, and cardiac myocyte growth. PI3K is divided into three classes; type I PI3K is preferentially expressed in leukocytes and activated by ${\beta}{\gamma}$ subunits of heterotrimeric G-proteins. In this study, the cDNAs encoding the $PI3K{\gamma}$ gene were isolated from a brain cDNA library constructed using the flounder (Paralichthys olivaceus). The sequence of the isolated $PI3K{\gamma}$ was 1341 bp, encoding 447 amino acids. The nucleotide sequence of the $PI3K{\gamma}$ gene was analyzed with that of other species, including Oreochromis niloticus and Danio rerio, and it turned out to be well conserved during evolution. The $PI3K{\gamma}$ gene was subcloned into the expression vector pET-44a(+), and expressed in the E. coli BL21 (DE3) codon plus cell. The resulting protein was expressed as a fusion protein of approximately 49 kDa containing a C-terminal six-histidine extension that was derived from the expression vector. The expressed protein was purified to homogeneity by His-tag affinity chromatography and showed enzymatic activity corresponding to $PI3K{\gamma}$. The binding of wortmannin to $PI3K{\gamma}$, as detected by anti-wortmannin antisera, closely followed the inhibition of the kinase activities. The results obtained from this study will provide a wider base of knowledge on the primary structure and characterization of the $PI3K{\gamma}$ at the molecular level.