• Tuberculosis and Respiratory Diseases
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• v.59 no.2
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• pp.186-192
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• 2005

Background : Diagnosis of tuberculous pleurisy is sometimes difficult using conventional diagnostic methods. We have investigated the utility of pleural fluid cell $IFN-{\gamma}$ production assay in the diagnosis of tuberculous pleurisy. Methods : We prospectively performed pleural fluid cell $IFN-{\gamma}$ production assay in 39 patients with tuberculous pleural effusions (TPE) and in 26 patients with nontuberculous pleural effusions (NTPE) (13 malignant pleural effusions and 13 parapneumonic effusions). Pleural fluid cells were cultured in DMEM media and stimulated with purified protein derivatives (PPD), and phytohemagglutinin (PHA) for 24 hr. The amount of $IFN-{\gamma}$ released in the culture supernatant was quantitated by $IFN-{\gamma}$ ELISA assay. We have also measured adenosine deaminase (ADA) activities and $IFN-{\gamma}$ concentrations in the pleural fluid. Results : 1) The pleural fluid levels of ADA activity and $IFN-{\gamma}$ concentrations were significantly higher in TPE than NTPE (p<0.01). 2) $IFN-{\gamma}$ production in TPE cells stimulated by PPD ($755,266{\pm}886,636pg/ml$) was significantly higher than NTPE cells ($3,509{\pm}6,980pg/ml$) (p<0.01). By considering the fact that $IFN-{\gamma}$ concentrations over 10,000 pg/ml is a criteria for the diagnosis of TBE, sensitivity and specificity of the test were 97.4 and 92.3%, respectively. 3) The ratios of $IFN-{\gamma}$ production by the stimulation with PPD and PHA (PPD/PHA) were significantly higher in TPE cells ($59{\pm}85$) than NTPE cells ($5{\pm}18$)(p<0.01). Considering the criteria for the diagnosis of TBE as PPD/PHA ratio over 5, sensitivity and specificity of the test were 76.9 and 92.3%, respectively. Conclusion : Pleural fluid cell $IFN-{\gamma}$ production assay may be useful for the diagnosis of tuberculous pleurisy.

• The Korean Journal of Pharmacology
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• v.9 no.1
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• pp.1-21
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• 1973

During the last decade extensive studios on catecholamines have evolved new knowledge in the physiology and biochemistry of adrenergic mechanism. Cardiac muscle, receiving adrenergic fibres from the stellate, cervical and thoracic ganglia, has been repeatedly shown to have a specific capacity to uptake and to store catecholamines. The catecholamine stores in cardiac muscle have also been shown to be important sites for the action of numerous drugs. Under normal condition, a certain level of catecholamines is maintained in the stores and serves as the basis for studying the changes in the catecholamine content of the heart. Because myocardial catecholamines play such important role in the patho-physiology of the heart, it would be interesting to compare the normal level of myocardial catecholamines among various species of animals. An occasional study has dealt with myocardial catecholamines of several species add ages of animals but these have been insufficiently comprehensive to afford a basis for an understanding of the importance of these amines as related to species and ages. The present investigation was undertaken to determine whether or not there is any significance of myocardial catecholamines in the course of the evolution and development of animals. Seasonal changes, sex difference and regional and subcellular distribution of myocardial catecholamines were also examined. The concentration of cardiac catecholamines was determined by the spectrophotofluorometric procedure described by Shore and Olin. The results obtained were summarized as follows: 1. As animals phylogenetically progressed larger amounts of catecholamines were resent in their hearts. A negligibly small amount of catecholamine was present in the hearts of the clam, a non-vertebrate. Among the vertebrates, cold-blooded animals (snake, turtle, frog, eel and fish) had less myocardial catecholamines than warm-blooded animals, of which aves (fowl and duck) had less than mammalia (cat, dog, rabbit, rat, cow and pig). The ratio of norepinephrine to epinephrine also was greater as the animals progress phylogenetically. 2. Examination of the regional distribution of cardiac catecholamines in warm-blooded animals showed that the content of the auricle was generally higher than that of the septum and considerably than that of the ventricle, but the differences of contents among these regions were not so marked. 3. In the embryonic chick, cardiac catecholamines were firstly detected on the 4th day of incubation, the time before the cardiac innervation of sympathetic nerves. The concentrations of these catecholamines increased but not markedly on the 6th day of incubation, soon after the innervation of sympathetic nerves to the heart. The level of the cardiac catecholamines fluctuated throughout the remainder of embryonic development. 4. In newborn rat hearts, a considerable amount of catecholamines was present. With the development of the rats, the concentrations of myocardial catecholamines increased. The ratio of epinephrine and norepinephrine fluctuated within the range of 40 to 60 pervent. However, as development progressed, the percentage of norepinephrine continued to rise, attaining the adult value of $80{\sim}90%$ after $45{\sim}60$ days. In contrast, the total amount of epinephrine remained fairly constant throughout the animal's development. 5. No significant sexual differences were observed in the concentration of myocardial catecholamines in the developing rat. 6. The catecholamines in the rabbit hearts increased during the summer season (from May to August) and maintained a fairly constant level in the other seasons of the year. 7. The subcellular distribution of cardiac catecholamines was examined by differential centrifugation of homogenates of cardiac muscles in rabbits, cats and rats. The catecholamines were found to be present approximately 20% in particles of mitochondrial fraction, 45% in particles of microsomal fraction and 35% in soluble supernatant fraction. The particle containing catecholamines in cardiac muscle appears to be two different sizes.

• Applied Biological Chemistry
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• v.10
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• pp.15-21
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• 1968

In order to fractionate the rice protein employing paper electrophoresis, 9 subjects of Korean rice and one Indica type, Pin Galw 50 were examined, the results were as follows. 1. Polished rice protein was separated into albumin, globulin, prolamine, and oryzenin. The amount of these fractions was determined by Kjeldahl method showing respectively 0.26%, 0.65%, 0.41%, and 5.01% in average. Albumin was extracted with deionized water, globulin with 10% NaCl, prolamine with 70% ethanol, and oryzenin with 0.05N-NaOH. 2. Albumin was extracted with deionized water and dialyzed by a cellophan tube. The supernatant was submitted to paper electrophoresis using phosphate buffer (pH 7.6, ${\mu}$ 0.18). Albumin was identified as monocomponent in all of 10 varieties under study. Globulin was extracted and dialyzed to remove the albumin. The precipitates were resolved in 10% saline solution and examined by paper electrophoresis. The globulin consists of two components in phosphate buffer(pH 7.6, ${\mu}$ 0.18) Any subject, regardless the origin, appears to contain globulin I and globulin II. Prolamine was extracted with 70% ethanol, dialayzed against deionized water, resolved with ethanol, and analyzed by Paper electrophoresis. It was proved as one component in the 70% alcoholic buffer(pH 9.0, ${\mu}$=0.0095). On the contrary, paper electrophoresis with oryzenin demonstrated two or three components in Sorensen's buffer(pH 13.0, ${\mu}$ 0.11). Yookoo 132, Dungpan 5, Kwansan, and Jaekun contain oryzenin I, oryzenin II, and oryzenin III. On the other hand, Paldal, Jinheung, Sukwang, Eunbangzu, Damakum, and Pin Galw 56 contain only oryzenin II, and oryzenin III. On the basis of these analyses a discussion of the differences between the protein fractions of 10 varieties was presented. 3. Globulin I varied from 0.22% to 0.46% (aver. 0.35%) in the amount, globulin II from 0.21 to 0.44%(aver. 0.32%), oryzenin I from 0.17% to 0.44%(aver. 0.3%), oryzenin II from 1.59% to 2.88%(aver. 2.23%), and oryzenin III from 2.02% to 3.57%(aver. 2.66%).

• Tuberculosis and Respiratory Diseases
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• v.48 no.2
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• pp.223-235
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• 2000

Background: Liquid ventilation is associated with decreased inflammatory response in an injured lung. This study was performed to investigate if whether perfluorocarbon(PFC) can decrease chemokine expression in airway epithelial cells. Methods: A549 cells were used for airway epithelial cells and perfluorodecalin for PFC. To expose cells to PFC, lower chamber of Transwell$^{(R)}$plate was used. This study was performed in two parts. In the first part, we examined whether PFC could decrease chemokine expression in airway epithelial cells through inhibition of other inflammatory cells. Peripheral blood mononuclear cells(PBMC's) were isolated and stimulated with lipopolysaccharide(LPS, 10 ${\mu}g/mL$) for 24 hours with or without exposure to PFC. Then A549 cells were stimulated with conditioned media(CM) containing the culture supernatants of PBMC. After 24 hours, the expressions of interleukin-8(IL-8) and RANTES were measured. In the second part of the study, we studied whether PFC could directly suppress chemokine expression in airway epithelial cells. A549 cells were stimulated for 24 hours with interleukin-l$\beta$ and/or tumor necrosis factor-$\alpha$ with or without exposure to PFC, and then the chemokine expression was measured. Northern analysis was used to measure the mRNA expression, and ELISA was used for immunoreactive protein measurements in culture supernatant. Results: 1. IL-8 and RANTES mRNA expression and immunoreactive protein production were increased significantly by CM from LPS-stimulated PBMC in A459 cells compared to with CM from unstimulated PBCM (p<0.05), but exposure of PFC had no significant effect on either mRNA expression or immunoreactive protein expression. 2. IL-8 and RANTES mRNA expression and immunoreactive protein production were increased significantly by IL-1$\beta$ and TNF-$\alpha$ in A549 cells(p<0.05), but exposure of PFC had no significant effect on neither either mRNA expression nor immunoreactive protein production. Conclusion : Decreased chemokine expression of airway epithelial cells may not be involved in decreased inflammatory response observed in liquid ventilation. Further studies on possible mechanisms of decreased inflammatory response are warranted.

• Journal of Life Science
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• v.26 no.5
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• pp.509-518
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• 2016

Asparagus cochinchinensis is a medical plant that has long been used to treat fever, cough, kidney disease, breast cancer, inflammatory disease and brain disease in northeast Asian countries. Although several studies have been conducted on the anti-neuroinflammatory effects of A. cochinchinensis, the correlation between these effects and nerve growth factor (NGF) has not yet been examined. In this study, we investigated the effects of an aqueous extract of A. cochinchinensis (AEAC) on the secretion and action mechanism of NGF in neuronal cells. The concentration of the NGF protein in the supernatant collected from cultured cells increased significantly in B35 cells treated with AEAC in comparison with the vehicle-treated group without any specific cytotoxicity. Furthermore, the mRNA expression of NGF showed a very similar pattern to its protein concentration. To examine the bioactivity of NGF secreted from B35 cells, undifferentiated PC12 cells were cultured in an AEAC-conditioned medium and neuritic outgrowth was observed. The dendrite length of PC12 cells in the AEAC-treated group was significantly higher than that in the vehicle-treated group. Moreover, the level of the downstream effectors p-TrkA and p-ERK of the high-affinity NGF receptor was significantly higher in the AEAC-treated group, while the expression of the downstream effectors of the low-affinity NGF receptor was significantly lower in the same group. These results suggest that AEAC may contribute to the regulation of NGF expression and secretion in neuronal cells; it is therefore an excellent candidate for further investigation as a therapeutic drug for neurodegenerative diseases.

• The Korean Journal of Nuclear Medicine
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• v.37 no.6
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• pp.402-417
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• 2003

Purpose: To evaluate the use of monoclonal antibody (MoAb) as a carrier of the receptor-binding ligand the receptor mediated uptake into liver and subsequent metabolism of $^{111}In-labeled$ galactosylated MoAb-chelator conjugates were investigated and compared with those of $^{111}In$ labeled MoAb. Materials and Methods : T101 MoAb, $IgG_2$ against human lymphocytic leukemic cell, conjugated with cyclic DTPA dianhydride (DTPA) or 2-p-isothiocyanatobenzyl-6-methyl-DTPA (1B4M) was galactosylated with 2-imino-2-methoxyethyl-1-thio-${\beta}$-D-galactose and then radiolabeled with $^{111}In$. Biodistribution and metabolism study was peformed with two $^{111}In-conjugates$ in mice and rats. Results: $^{111}In-labeled$ T101 and its galactosylated conjugates were taken to the liver by the time, mostly within 10 min. However DTPA conjugate was retained longer in the liver than the 1B4M conjugate (55% vs 20% of injected dose at 44 hr). During this time, the radiornetabolite of DTPA conjugate was excreted similarly into urine (24%) and feces (17%). The radiometabolite of 1B4M was excreted primarily into feces (68%) rather than urine (8%). Size exclusion HPLC analysis of the bile and supernatant of liver homogenate showed two peaks the first (35%) with the retention time (Rt) identical to IgG and the second (65%) with Rt similar to free $^{111}In$ at 3 hr post-injection for the 1B4M conjugate, indicating that the metabolite is rapidly excreted through the biliary system. in contrast to DTPA conjugate, the small $^{111}In-DTPA-like$ metabolite was the major radioindium component (90%) in the liver homogenate as early as 3 hour post-injection, but the cumulative radioindium activity in feces was only 17% at 44 hour, indicating that the metabolite from DTPA conjugate does not clear readily through the biliary tract. Conclusion: The galactosylation of the MoAb conjugates resulted in higher hepatocyte uptake and enhanced metabolism, compared to those without galactosylation. Metabolism of the MoAb-conjugates is different between compounds radiolabled with different chelators due to different characteristics of radiometabolites generated in the liver.

• Journal of the Korean Society of Food Science and Nutrition
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• v.39 no.1
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• pp.31-35
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• 2010

This study was designed to evaluate anti-diabetes effect of Helianthus tuberosus extract (HT) in HIT-T15 cells. There were 5 experimental groups according to treatment NC (0 ${\muL/mL$), HT2 (1.1 ${\muL/mL$), HT3 (1.5 ${\muL/mL$), IN2 (1.8 ${\muL/mL$), IN3 (2.5 ${\muL/mL$). Inulin (IN) was used as a positive control for the Helianthus tuberosus extract groups. Cell viability was significantly increased in the HT3 (1.5 ${\muL/mL$), IN2 (1.8 ${\muL/mL$), IN3 (2.5 ${\muL/mL$) groups, compared with the NC group. There was no significant difference in cytotoxicity among all groups. Cell survival by MTT assay with alloxan was significantly increased in the HT2 (1.1 ${\muL/mL$), HT3 (1.5 ${\muL/mL$) groups, compared with the NC group. Insulin secretion and NAD+/NADH ratio were significantly increased in the HT3 group, compared with the NC group. We found that Helianthus tuberosus extract increased cell viability, had a protective effect on $\beta$-cells, and increased insulin secretion level and $NAD^+$/NADH ratio in HIT-T15 cells. These results suggest that Helianthus tuberosus extract improves the diabetes-related factors.

• Clinical and Experimental Pediatrics
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• v.51 no.10
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• pp.1096-1101
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• 2008

Purpose : Patients with chronic granulomatous disease (CGD) have genetic mutations in a component of the NADPH oxidase enzyme that is necessary for the generation of the superoxide anion. The profound defect in innate immunity is reflected by the patients susceptibility to catalase-positive bacteria and fungi. In addition, CGD patients display signs of persistent inflammation, which is not associated only with deficient superoxide anion production. The aim of this study was to elucidate the cytokine responses in CGD patients after $TNF-{\alpha}$ stimulation. Methods : Heparinized blood samples were collected from 8 CGD patients and 10 healthy volunteers. Monocytes ($1{\times}10^6cell/well$) isolated by the magnet cell isolation system were incubated with a constant amount of $TNF-{\alpha}$ (10 ng/mL) at $37^{\circ}C$ for 6 h. Incubated cells were harvested at 60-min intervals for IL-8 and IL-10 mRNA analysis, and the supernatant was collected at the same intervals to determine IL-8 and IL-10 expression. Monocytes from healthy volunteers were also incubated with antioxidants followed by $TNF-{\alpha}$ stimulation for IL-8 and IL-10 expression. Results : In CGD patients, a high expression of IL-8 together with a significantly higher IL-10 expression than in the healthy controls was seen after $TNF-{\alpha}$ stimulation. Moreover, normal monocytes treated with antioxidants exhibited increased IL-8 responses. Conclusion : The absence of phagocyte-derived reactive oxidants in CGD might be associated with a dysregulated production of pro- and antiinflammatory cytokines. Additional research related to reactive oxidants is needed to clarify the role of cytokines in CGD patients.

• Microbiology and Biotechnology Letters
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• v.37 no.1
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• pp.33-41
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• 2009

The crushed fruiting body of Alaskan Porodaedalea pini (Brot.) Murrill (syn. Phellinus pini) was extracted in boiling water for 4 h with the yield of 20.5% in dry mass. This hot-water extract showed significant antiviral activity by inhibiting the plaque formation in HeLa cells by coxackievirus B3 (CVB3) and also showed highest inhibitory effect against neuraminidase activity among water extracts of various mushrooms. From the water extract, the ethanol precipitate (EP) and supernatant fraction (ES) were obtained through 75% ethanol precipitation with the yield of 43.3% and 28.3% in dry mass, respectively. Whereas ES did not show any detectable level of antiviral activity, EP showed significant dose-dependent inhibition of plaque formation by CVB3 in HeLa cells with an $EC_{50}$ (50% effective concentration) of 0.45 mg/mL. The cytotoxicity on HeLa cells by EP was relatively low with the $CC_{50}$ (50% cytotoxic concentration) of 2.25 mg/mL. EP also effectively inhibited neuraminidase activity in a dose-dependent manner showing up to 75% inhibition at 1.7 mg/mL. These results suggest that the hot-water extract and its EP of P. pini fruiting body can be a candidate for the development of a potent broad-range antiviral agent against influenza virus(Flu) as well as CVB3. The major active component of EP was shown to be a heteropolysaccharide-protein complex containing glucose as the main sugar residue with mole percentage of 79.8% and other sugars like galactose (19.2%), xylose (17%), mannose (5.8%), and fucose (4.6%) and a small portion (12.7%, in mass) of protein.

• Journal of Animal Science and Technology
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• v.48 no.6
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• pp.907-920
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• 2006

This study was conducted to investigate the inhibitory activity of Lactobacillus spp., Bacillus ssp., and calf fecal isolates against pathogenic Salmonella typhimurium, E. coli, Listeria monocytogenes, and Staphylococcus aureus. Among thirteen strains of Lactobacillus ssp. tested, Lactobacillus helveticus CU631 showed the highest inhibition against three pathogens, whereas Bacillus spp. showed a weak inhibitory activity. Four calf fecal isolates were identified as Lactobacillus pentosus CU13, CU05, Pediococcus pentosaceus CUR02, and Lactobacillus lactis ssp. lactis CUM14. The whole cell and cell wall components of L. rhamnosus CU02 and L. pentosus CU13 were active in the inhibition of L. monocytogenes. The medium components and levels, which affect on the inhibitory activity, were revealed as Tween 80 1.0%, peptone 3.0%, yeast extract 3.0%, glucose 3.0%, beef extract 3.0%, and NaCl 1.0～3.0%, respectively. Inhibitory activity of the supernatant culture medium was not affected by catalase and proteinase K treatment but affected by heat treatment at 80℃ and netralization, which implies that the inhibitory activity is due to the production of organic acids during the growth. L. pentosus CU13 and L. rhamnosus CU02 exhibited broad inhibition spectrum against 16 out of 21 strains including some pathogens. Oral administration of L. rhamnosus CU02 to the mice infected with E. coli O157:H7 was proven to be effective to recover their body weight during the experimental period.