Background : Mucus hypersecretion in the patients with airway diseases represents poor prognosis as well as discomfort. However, there is no known therapy for its effective control. One important component of mucus is mucin, a glycosylated protein, which endows mucus with viscosity. We studied whether a proteinase has a role in mucin secretion and if so, which. Methods : (1) Inhibition of mucin secretion Group-specific proteinase inhibitors were tested to evaluate whether a proteinase belonging to a group of proteinases plays a role in mucin secretion. Phenylmethylsulfonyl fluoride(PMSF, a serine proteinase inhibitor), E-64(a cysteine proteinase inhibitor), Pepstatin(an aspartic proteinase inhibitor) and 1, 10-Phenanthroline(a metalloproteinase inhibitor) were treated into the Calu-3 cell line for 24 hours. The enzyme linked immunoabsorbant assay(ELISA) for MUC5AC was performed to evaluate the amount of mucin secretion and to compare with a control. (2) Stimulation of mucin secretion Matrix metalloproteinase-9(MMP-9), MMP-12 and TACE(TNF-alpha converting enzyme), which are known to be related with airway diseases, were used to be treated into Calu-3 for 24 hours. ELISA for MUC5AC was performed to evaluate the amount of mucin secretion and to compare with the controls. Results : (1) PMSF($10^{-4}M$), E-64($10^{-4}M$), Pepstatin($10^{-6}M$) and 1, 10-Phenanthroline($10^{-4}M$) reduced the MUC5AC secretion by $1{\pm}4.9%$(mean${\pm}$standard deviation; P=1.0 compared with the control), $-6{\pm}3.9%$(P=0.34), $-13{\pm}9.7%$(P=0.34) and $41{\pm}8.2%$(P=0.03), respectively. (2) The amounts of MUC5AC secretion stimulated by MMP-9(250ng/ml), MMP-12(100ng/ml) and TACE(200ng/ml) were $103{\pm}6%$(P=0.39), $102{\pm}8%$(P=1.0) and $107{\pm}13%$(P=0.39), respectively, compared with the controls. Conclusion : Metalloproteinase(s) is (are) suggested to play a role in mucin secretion. It appears that metalloproteinases, other than MMP-9, MMP-12 or TACE, affect the mucin secretion in this in vitro model.
Sox4, a transcription factor, consists of three functional domains: an HMG-box domain as a DNA binding domain, serine rich region as a transactivation domain and glycine rich region (GRR), an unknown functional domain. Although Sox4 is known to be functionally involved in heart, B-cell and reproductive system development, its physiological function remains to be elucidated. We used pGEX expression system to develop a simple and rapid method for purifying Sox4 protein in suitable forms for biochemical studies of their functions. Unexpectedly, we observed that full-length Sox4 appears to be protease-sensitive during expression and purification in E. coli. To map the protease-sensitive site in Sox4, we generated various constructs with each of functional domains of Sox4 and purified as the GST-Sox4 fusion proteins using glutathione beads. We found that the specific cleavage site for the proteolytic enzyme, which exists in E. coli, is localized within the novel GRR of Sox4. Our study suggest that the GRR of Sox4 may a target for the cellular protease action and this cleavage in the GRR may be involved in regulating physiological function of Sox4. Additionally, our study may provide a useful method for investigating the proteolytic cleavage of the target molecule in E. coli.
In order to develope a new kind of fermented milk, basic studies on several lactic acid bacteria and yeasts were conducted, 8 kinds of alcohol fermented milk were manufactured and sensory evaluation was undertaken. The results obtained are summarized as follows: 1. Four kinds of lactic acid bacteria were isolated, among which Y-2 strain was strongest in acid productivity and it was elucidated that acid productivity of all strains was stornger in synthetic medium than in milk medium. 2. The pH in milk medium inoculated with Y-2 strain and incubated at $30^{\circ}C$ for 24 hours was dropped from 5.8 to 3.8 and fluctuation in amino nitrogen content was found during incubation. 3. The pH in milk medium inoculated with K. fragilis and incubated at $30^{\circ}C$ for 7 days was dropped from 6.2 to 5.2 and amino nitrogen content was in the range of $0.12{\sim}0.27mg/ml$. Alcohol productivity of K. fragilis was stronger than E-2 and E-4 strain but no difference in alcohol productivity was found between milk medium and synthetic medium. 4. The repression in growth and acid productivity of lactic acid bacteria was recongnized if inoculated after inoculating yeast firstly. 5. Alcohol productivity was increased rapidly at the end of acid production of lactic acid bacteria if lactic acid bacteria if lactic acid bacteria and yeast were inoculated simultaneously. 6. Sensory evaluation showed that the product that alcohol content and acidity were 1% and 0.8% respectively had the best palatability(p<0.01). 7. Chemical composition of final product was similar to that of milk koumiss in ash, protein and moisture content.
Jang, Su Kil;Ahn, Jeong Won;Jo, Boram;Kim, Hyun Soo;Kim, Seo Jin;Sung, Eun Ah;Lee, Do Ik;Park, Hee Yong;Jin, Duk Hee;Joo, Seong Soo
Korean Journal of Food Science and Technology
/
v.51
no.2
/
pp.160-168
/
2019
This study aimed to determine whether double-processed ginseng berry extract (PGBC) could improve learning and memory in an $A\hat{a}42$-induced Alzheimer's mouse model. Passive avoidance test (PAT) and Morris water-maze test (MWMT) were performed after mice were treated with PGBC, followed by acetylcholine (ACh) measurement and glial fibrillary acidic protein (GFAP) detection for brain damage. Furthermore, acetylcholinesterase (AChE) activity and choline acetyltransferase (ChAT) expression were analyzed using Ellman's and qPCR assays, respectively. Results demonstrated that PGBC contained a high amount of ginsenosides (Re, Rd, and Rg3), which are responsible for the clearance of $A{\hat{a}} 42$. They also helped to significantly improve PAT and MWMT performance in the $A{\hat{a}} 42-induced$ Alzheimer's mouse model when compared to the normal group. Interestingly, ACh and ChAT were remarkably upregulated and AChE activities were significantly inhibited, suggesting PGBC to be a palliative adjuvant for treating Alzheimer's disease. Altogether, PGBC was found to play a positive role in improving cognitive abilities. Thus, it could be a new alternative solution for alleviating Alzheimer's disease symptoms.
Hwa Sin Lee;Bo Bae Park;Sun Nyoung Yu;Min Ji Kim;Yun Jin Bae;Yi Rooney Lee;Ye Eun Lee;Si Yoon Kim;Yun Ho Shim;Soon Cheol Ahn
Journal of Life Science
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v.33
no.10
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pp.783-790
/
2023
Modern people have an increased incidence of metabolic diseases due to changed eating habits, and diabetes is considered the most significant metabolic disease. Given that existing diabetes treatments are accompanied by side effects, the aim of this study was to identify traditional natural products that have anti-diabetic activity. The potential anti-diabetic and antioxidant activities of natural products were examined using 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging assay, α-glucosidase assay, and protein tyrosine phosphatase 1B (PTP1B) inhibition assay. Methanol extracts of Ulmus davidiana var. japonica, Acer tegmentosum branches, Nelumbo nucifera seeds, and Carthamus tinctorius seeds were found to have high anti-diabetic activity and further fractionated with solvents using ethyl acetate and butanol. Consequently, the ethyl acetate fraction of C. tinctorius seeds (MG-11-E) with high α-glucosidase and PTP1B inhibitory activity was selected. MG-11-E was subjected to preparative thin layer chromatography, and fraction #6 showed high α-glucosidase and PTP1B inhibitory activity. Fraction #6 was analyzed and fractionated via high performance liquid chromatography with 50% methanol as the mobile phase, and anti-diabetic activity was observed in the sample that eluted after 4 min as a single peak. The α-glucosidase inhibitory activity exhibited by this sample seemed to be greater than the PTP1B inhibitory activity; thus, it was concluded that a greater anti-diabetic therapeutic effect may be achieved by combining this agent with natural products that inhibit PTP1B activity.
Kim, So Ri;Choe, Yeong Hun;Lee, Ka Young;Min, Kyung Hoon;Park, Seoung Ju;Lee, Heung Bum;Lee, Yong Chul;Rhee, Yang Keun
Tuberculosis and Respiratory Diseases
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v.64
no.2
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pp.125-132
/
2008
Background: In chronic obstructive pulmonary disease (COPD) patients, the serum levels of C-reactive protein (CRP) are elevated and an increase of CRP is more exaggerated in the acute exacerbation form of COPD (AECOPD) than in stable COPD. Pulmonary arterial hypertension is a common complication of COPD. An increased level of CRP is known to be associated with the risk of systemic cardio-vascular disorders. However, few findings are available on the potential role of CRP in pulmonary arterial hypertension due to COPD. Methods: This study was performed prospectively and the study population was composed of 72 patients that were hospitalized due to AECOPD. After receiving acute management for AECOPD, serum CRP levels were evaluated, arterial oxygen pressure ($PaO_2$), was measured, and the existence of pulmonary arterial hypertension under room air inhalation was determined in the patients. Results: The number of patients with pulmonary arterial hypertension was 47 (65.3%)., There was an increased prevalence of pulmonary arterial hypertension and an increase of serum CRP levels in patients with the higher stages of COPD (e.g., patients with stage 3 and stage 4 disease; P<0.05). The mean serum CRP levels of patients with pulmonary arterial hypertension and without pulmonary arterial hypertension were $37.6{\pm}7.4mg/L$ and $19.9{\pm}6.6mg/L$, respectively (P<0.05). However, there was no significant difference of the mean values of $PaO_2$ between patients with pulmonary arterial hypertension and without pulmonary arterial hypertension statistically ($77.8{\pm}3.6mmHg$ versus $87.2{\pm}6.0mmHg). Conclusion: We conclude that higher serum levels of CRP can be a sign for pulmonary arterial hypertension in AECOPD patients.
Journal of the Korean Society of Food Science and Nutrition
/
v.40
no.6
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pp.767-774
/
2011
Defatted green tea seed was extracted with 100% ethanol for 4 hr and then fractionated with petroleum ether, ethyl acetate and butanol. The ethanol and butanol extracts showed greater increases in antiproliferation potential against liver cancer cells than petroleum ether, ethyl acetate, $H_2O$, and hot water extracts did. Thus, this study was carried out to investigate the anti-proliferative actions of defatted green tea seed ethanol extract (DGTSE) in HepG2 cancer cells. The DGTSE contained catechins including EGC ($1039.1{\pm}15.2\;g/g$), tannic acid ($683.5{\pm}17.61\;{\mu}g/g$), EC ($62.4{\pm}5.00\;{\mu}g/g$), ECG ($24.4{\pm}7.81\;{\mu}g/g$), EGCG ($20.9{\pm}0.96\;{\mu}g/g$) and gallic acid ($2.4{\pm}0.68\;{\mu}g/g$), but caffeic acid was not detected when analyzed by HPLC. The anti-proliferation effect of DGTSE toward HepG2 cells was 83.13% when treated at $10\;{\mu}g$/mL, of DGTSE, offering an $IC_{50}$ of $6.58\;{\mu}g$/mL. DGTSE decreased CYP1A1 and CYP1A2 protein expressions in a dose-dependent manner. Quinone reductase and antioxidant response element (ARE)-luciferase activities were increased about 2.6 and 1.94-fold at a concentration of $20\;{\mu}g$/mL compared to a control group, respectively. Enhancement of phase II enzyme activity by DGTSE was shown to be mediated via interaction with ARE sequences in genes encoding the phase enzymes. DGTSE significantly (p<0.05) suppressed prostaglandin $E_2$ level, tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) protein expressions, and NF${\kappa}$B translocation, but did not affected nitric oxide production. From the above results, it is concluded that DGTSE may ameliorate tumor and inflammatory reactions through the elevation of phase II enzyme activities and suppression of NF${\kappa}$B translocation and TNF-${\alpha}$ protein expressions, which support the cancer cell anti-proliferative effects of DGTSE in HepG2 cells.
Purpose : We evaluated clinical manifestations and laboratory findings in patients with measles according to C-reactive protein(CRP) concentration. Methods : A retrospective analysis was performed using the medical records of patients with measles at The Catholic University of Korea, Daejeon St. Mary's Hospital from October 1999 to May 2000. We divided the patients with measles into four groups according to CRP level, i.e., those with below 5 mg/L(134 patients, negative group), those with 6-19 mg/L(85 patients), those with 20-49 mg/L(27 patients), and those over 50 mg/L(7 patients). We compared clinical and laboratory characteristics among these four groups. Results : The mean CRP level of all patients was $11.1{\pm}7.5mg/L$. No statistical differences were present between the negative group and the 6-19 mg/L group or the 20-49 mg/L group in the duration of fever, hospitalization days, complications determined with longer hospitalization for more than eight days, white blood cell count, and incidence of hepatitis. Compared with the negative group, the over 50 mg/L group showed a longer duration of fever($4.7{\pm}1.7$ vs $7.2{\pm}3.9days$), duration of hospitalization($5.4{\pm}1.4$ vs $9.4{\pm}4.7days$), incidence of complications(5.2% vs 42.9%) and a higher mean level of WBC count($5,900{\pm}2,700/mm^3$ vs $12,700{\pm}6,700/mm^3$). With an increasing CRP level, there was a tendency for the duration of fever, complications and WBC count to increase. However the levels of liver enzymes(AST/ALT) were not associated with CRP level. Conclusion : A CRP level of over 50 mg/L in measles is associated with severity and complications.
The nutritional status is strictly related with flood production, flood processing, and distribution along with habits, education and technological achievement, adapted and adjusted to socio-economic conditions. All these factors are independently affecting the nutritional status of populations. In addition to the above mentioned factors launch of it is useful to consider two points ; unification of the South and the North Korea and WTO. The present study gives and overview of basic knowledge about nutritional status of Korean by using availab1e data in relation to nutrition. The basic characteristics of Korean diet with proportions high in carbohydrate and low in fat, have been relatively constant for the past two decades. The average daily flood intake in terms of weight of flood per person is relatively constant throughout the years. Although the proportion of animal food intake tends to increase recently, the Korean diet is still insufficient in meat, eggs, milk, and fish. Moreover because milk has not been accustomed flood with the general population and not much used in traditional flood preparation in Korea, milk consumption was especially low in comparison with western countries. The total energy intake was relatively constant throughout the years from 1969 to 1993. However, changes in the composition have occurred in the past two decades. The amount and proportions of fat have been s1ightly increasing while the total amount of carbohydrate has been decreasing. The nutrition preblems of Korea have changed over the past severa1 decades. The general adequacy of protein and energy existing after Korean War(1950) was resolved now. Since then the average diet appears to be nearly desirable in terms of rapid rates of growth during childhood and attainment of progressively stature and body weight at maturity. The dietary habits of some young people in these days seem to be taking a more western style diets. This trends if established by habit may lead to a marked change in the traditional diet and health. I think Korean nutritional experiences have potential values for tole other countries in Asia and in western countries. Korean diet illustrates a high level of nutritional status and health attainable will a largely vegetable diet : high in complex carbohydrates, and dietary fibre, and low in tat, and reasonable amount of total protein. This is significant for developing and developed countries that must select specific goals fir adequate nutrition for the people. Compared to the western countries, Koreas different incidence of coronary heart disease and malignancy demonstrates the significance of environment and probably the prominent role of diet in the development of these diseases. The changes occurring in the Korean diet of fir the unusual opportunity to assess the effect of diet upon chronic degenerative disease. In the future, the Korean diet might be continue to change significantly These changes are being influenced by socioeconomic factors that have been emerging and growing stronger since mid-l980 and that probably continue to be potent. The expanded purchasing power of the consumer results in increased discretionary purchases. In the case of foodstuffs, the consumer demands appear to be directed toward items of higher protein content, which, being primarily animal products, are inevitably accompanied by an increased consumption of fat. The continued availability of these more expensive flood items depends upon the balance of foreign trade favouring their importation and domestic production. A regression of foreign trade could result in a decreased supply for the consumer, whereas continued growth of trade iou]d provide freedom for increasing availability to the consumer. In this latter situation the exact choice of foodstuffs is depending upon comsumer tastes and the pressures that may influence it.
Park, Sang-Dong;Kim, Min-Jeong;Lee, A-Ram;Jang, Jun-Hyouk;Kim, Kyung-Ho
Journal of Acupuncture Research
/
v.19
no.2
/
pp.51-64
/
2002
We have compared(using the same series of experimental tissue samples) the levels of proteolytic enzyme activities and free radical-induced protein damage in synovial fluid from RA and CPH cases. Many protease types showed significantly increased (typically by a factor of approximately 2-3-fold) activity in RA, compared to normal rats. However, CPH significantly reduced the cytoplasmic enzyme activities of arginyl aminopeptidase, leucyl aminopeptidase, pyroglutamyl aminopeptidase, tripeptidyl aminopeptidase, and proline endopeptidase to almost about 1/10 each. For the Iysosomal proteases, synovial fluid samples from RA rats, CPH significantly reduced the enzyme activities of cathepsin B, dipeptidyl aminopeptidase I and dipeptidyl aminopeptidase II. In extracellular matrix degrading(collagenase, tissue elastase) and leukocyte as sociated proteases (leukocyte elastase, cathepsin G), CPH decreased these enzyme activities of collagenase, tissue elastase and leukocyte associated elastase in RA. In cytoplasmic and lysosomal protease activities in plasma from RA. CPH and normal plasma samples were not significantly different, suggesting that altered activity of plasma proteases (particularly those enzymes putatively involved in the immune response) is not a contributory factor in the pathogenesis of RA. In addition, the level of free radical induced damage to synovial fluid proteins was approximately twice that in RA, compared with CPH. CPH significantly decreased the level of ROS induced oxidative damage to synovial fluid proteins (quantified as protein carbonyl derivative). Therefore we conclude that both proteolytic enzymes and free radicals are likely to be of equal potential importance as damaging agents in the pathogenesis of inflammatory joint disease, and that the design of novel therapeutic strategies for patients with the latter disorder should include both protease inhibitory and free radical scavenging elements. In addition, the protease inhibitory element should be designed to inhibit the action of a broad range of protease mechanistic types (i.e. cysteine-, metallo- and serine- proteinases and peptidases). However, increased protein damage induced by ROS could not be rationalised in terms of compromised antioxidant total capacity, since the latter was not significantly altered in RA synovial fluid or plasma compared with CPH.
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