• The Journal of Pediatrics of Korean Medicine
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• v.21 no.3
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• pp.33-55
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• 2007

Objectives In the present study, the author intended to investigate whether bojung-ikgitang-gamibang(BJGB) and seonbang-paedoktang(SBPT) significantly affect in vivo and in vitro mucin secretion from airway epithelial cells. Methods In vivo experiment, mice's mucin which is on a hypersecretion of airway mucin, mice's tracheal goblet cells in hyperplasia and mice's intraepithelial mucosubstances were exposed with SO2for3weeks. Effects of orally-administered BJGB and SBPT during 1 week on vivo mucin secretion and hyperplasia of tracheal goblet cells were assessed by using both enzyme-linked immunosorbent assay(ELISA) and staining goblet cells with alcian blue. In vitro experiment, confluent hamster tracheal surface epithelial(HTSE) cells were metabolically radiolabeled with 3H-glucosamine for 24hrs and chased for 30 min in the presence of each agent to figure out the effectiveness of 3H-mucin secretion. Total elution profiles of control spent media and treatment sample through Sepharose CL-4B column were analyzed. The effects of each agent on contractility of isolated tracheal smooth muscle and effects of each agent on MUC5AC gene expression in cultured HTSE cells were investigated. Also, possible cytotoxicities of each agent were assessed by measuring lactate dehydrogenase(LDH) release. Additionally, effects of BJGB and SBPT on both MUC5AC gene expression in cultured HTSE cells and TNF- or EGF-induced MUC5AC gene expression in human airway epithelial cells (NCI-H292) were investigated. Results (1) BJGB and SBPT inhibited hypersecretion of in vivo mucin. SBPT also inhibited the increase the number of goblet cells. However, BJGB did not affect the increase of number of goblet cells; (2) BJGB significantly increased mucin secretion from cultured HTSE cells, without significant cytotoxicity, and chiefly affected the 'mucin' secretion; (3) SBPT did not affect mucin secretion from cultured HTSE cells without significant cytotoxicity, and also did not affect the secretion of the other releseable glycoproteins; (4) BJGB and SBPT did not affect Ach-induced contraction of isolated tracheal smooth muscle; (5) SBPT significantly inhibit the expression levels of MUC5AC gene and BJGB significantly increased the expression levels of MUC5AC gene in both HTSE cells and NCI-H292 cells. Conclusions BJGB and SBPT can not only affect the secretion of mucin but also affect the expression of mucin gene. The author suggests that the effects BJGB and SBPT with their components should be further investigated and it is highly desirable to find from oriental medical prescriptions, novel agents which might regulate hypersecretion of mucin from airway epithelial cells.

• Molecular & Cellular Toxicology
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• v.2 no.3
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• pp.176-184
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• 2006

The controversy on genotoxicity of molinate, an herbicide, has been reported in bacterial system, and in vitro and in vivo mammalian systems. To clarify the genotoxicity of molinate, we performed bacterial gene mutation test, in vitro chromosome aberration and mouse lymphoma $tk^{+/-}$ gene assay, and in vivo micronucleus assay using bone marrow cells and peripheral reticulocytes of mice. In bacterial gene mutation assay, no mutagenicity of molinate ($12-185{\mu}g/plate$) was observed in Salmonella typhimurium TA 98, 100, 1535 and 1537 both in the absence and in the presence of S-9 metabolic activation system. The clastogenicity of molinate was observed in the presence ($102.1-408.2\;{\mu}g/mL$) of metabolic activation system in mammalian cell system using Chinese hamster lung fibroblast. However, no clastogenicity was observed in the absence ($13.6-54.3\;{\mu}g/mL$) of metabolic activation system. It is suggested that the genotoxicity of molinate was derived some metabolites by metabolic activation. Molinate was also subjected to mouse lymphoma L5178Y $tk^{+/-}$ cells using microtiter cloning technique. In the absence of S-9 mixture, mutation frequencies (MFs) were revealed $1.4-1.9{\times}10^{-4}$ with no statistical significance. However, MFs in the presence of metabolic activation system revealed $3.2-3.4{\times}10^{-4}$ with statistical significance (p<0.05). In vivo micronucleus (MN) assay using mouse bone marrow cells, molinate revealed genotoxic potential in the dose ranges of 100-398 mg/kg of molinate when administered orally. Molinate also subjected to acridine orange MN assay with mouse peripheral reticulocytes. The frequency of micronucleated reticulocytes (MNRETs) induced 48 hr after i.p. injection at a single dose of 91, 182 and 363 mg/kg of molinate was dose-dependently increased as $10.2{\pm}4.7,\;14.6{\pm}3.9\;and\;28.6{\pm}6.3\;(mean{\pm}SD\;of\;MNRETs/2,000\;reticulocytes)$ with statistical significance (p<0.05), respectively. Consequently, genotoxic potential of molinate was observed in in vitro mammalian mutagenicity systems only in the presence of metabolic activation system and in vivo MN assay using both bone marrow cells and peripheral reticulocytes in the dose ranges used in this experiment. These results suggest that metabolic activation plays a critical role to express the genotoxicity of molinate in in vitro and in vivo mammalian system.

• Journal of Embryo Transfer
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• v.24 no.3
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• pp.159-167
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• 2009

Ultrasonographic examination was performed to observe the ultrasonographic image of Korean native cows' normal uterus in condition of in vitro and in vivo. The experiment was done 28 slaughtered cows' uterus using immersed in water in vitro, and 41 healthy breeding cows taken rectal ultrasonography in vivo. Ultrasonographic examination of uterine was taken on the reference of cross section of intercornual ligaments' cranial. Each uterus on the experiments was compared by estrous cycle and ultrasonographic frequency. The uterine structure using ultrasonography was 5 layers of uterine horn in vivo as well as in vitro. Uterine horn was observed to be distinguished from inside to outside as endometrium to inner echogenic layer, circular muscle layer to slightly echogenic elliptical layer, stratum vasculare to central echogenic layer, longitudinal muscle layer to slightly echogenic arched layer, and perimetrium to outer echogenic layer, respectively. According to the observation of uterus related to estrous cycle and ultrasonographic examination, uterine endometrium in vitro was constantly founded irrespective of estrous cycle and ultrasonographic frequency. On the low frequency, endometrium and circular muscle layer in estrus were prone to distinguished than in diestrus. On the high frequency, endometrium and circular muscle layer were always distinguished regardless of estrous cycle. In vivo, uterine endometrium and circular muscle layer were observed regardless of estrus and ultrasonographic frequency. On the low frequency, stratum vasculare and longitudinal muscle layer were not likely to be distinguished in diestrus, but estrus. On the high frequency, stratum vasculare and longitudinal muscle layer were observed regardless of estrous cycle. Also, every uterine structure was easily distinguished on high frequency than low frequency owing to precision of distinction in layers. The difference of results followed by the experiments conditions between in vitro and in vivo was that uterine endometrium and circular muscle layer in diestrus in vitro were difficult to be distinguished and uterine lumen was observed during whole estrous cycle. In vivo, It was founded that the distinction of stratum vasculare and logitudinal muscle layer in diestrus was complicated and uterine lumen was observed during only estrus. In view of the result so far achieved, normal uterine structure divided in 5 layers on ultrasonography was accorded with microscopic organization, uterine structure was likely to be observed during estrus than diestrus, high frequency checkup than low frequency, and uterine endometrium, circular muscle, stratum vasculare was easily observed regardless of estrous cycle and ultrasonographic frequency.

• Asian-Australasian Journal of Animal Sciences
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• v.14 no.4
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• pp.496-500
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• 2001

In vitro and in vivo experiments were carried out to evaluate the nutritive value of three tropical browse species as assessed by DM and CP digestibility, and NDF and ADF degradability with incubation times (T) of 6, 12, 24 and 48h. During the in vivo digestibility experiment three male castrated sheep (age 16 - 25 months) with a mean liveweight of $11.5{\pm}0.9kg$ were placed in individual metabolism stalls and were allocated to one of the three browse species in a $3{\times}3$ Latin square design. The browse species were all leguminous and consisted of: Acacia sieberina (A. sieberina), Ficus polita (F. polita), and Ficus sycomorus (F. sycomorus). The mean DM and CP contents of F. polita were higher than for A. sieberina and F. sycomorus (p<0.05). In contrast the NDF and ADF contents of F. sycomorus were higher compared to the other species examined (p<0.05). The in vitro DM and CP digestibility, and NDF and ADF degradability observed at different stages of incubation were higher in F. polita followed by A. sieberina and F. sycomorus. The DM and CP digestibility at 48 h incubation were 72.92, 74.84 and 53.52% and 77.38, 77.68 and 63.64% for A. seiberina, F. polita and F. sycomorus, respectively. This shows that F. polita contains more soluble materials which ruminant can benefit from and hence has more feeding value. The fermentation of F. sycomorus was slower for all the nutrients evaluated due to the presence of more fibre. Similarly, higher in vivo digestibility coefficient of DM, CP, NDF, ADF and hemicellulose were observed for F. polita reflecting its higher values of CP, ether extract (EE) and hemicellulose associated with lower values of NDF and ADF. Higher DMI and daily gain were recorded in sheep during feeding of F. polita compared to the other species evaluated. The digestibility of all the nutrients examined were higher in the in vivo than in the in vitro trial except for CP and DM. Sheep showed no visual signs of toxicity throughout the study periods. These results showed A. seiberina, F. polita and F. sycomorus can sustain sheep on a maintenance diet and could as well be used as a supplementary feed to low producing animals during the tropical dry season. Further research is needed to ascertain the viability of using these browse species on a long-term basis.

• Journal of Pharmacopuncture
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• v.7 no.2
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• pp.43-56
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• 2004

Objectives : In order to investigate effects and immune improvement of distilled red-ginseng herbal Acupuncture, expression of Cox-1, Cox-2, and mRNA of Bcl-2 and Bax were analyzed in A549 cell in vivo. Survival time and expression of cytokine mRNA were measured for the mice with Sarcoma-180 induced abdominal cancer. Methods : Balb/c mouse was treated with distilled red-ginseng Herbal Acupuncture at Wisu($BL_{21}$) and Chung- wan($CV_{12}$) to investigate anti-cancer effects and immune response. Results : 1. For expression of mRNA of Cox-1 using RT-PCR, the control group and the experiment groups didn't show significant differences. For Cox-2, both experiment groups and the normal group showed significant differences. 2. For expression of mRNA of Bcl-2 using RT-PCR, experiment groups showed slight decrease compared to the control group. For Bax, no significant changes were shown between the control group and experiment groups. 3. For survival time, all of experiment groups showed 11.1% increase compared to the control group. 4. For IL-2 and IL-4 productivity using Flow cytometry, all of experiment groups didn't show any significance. 5. For IL-2 productivity using ELISA, all of experiment groups didn't show any significance. 6. For expression of cytokine mRNA using RT-PCR, significant increase of IL.-2 and IL-4 were witnessed in the experiment group II compared to the control group. Significant increase of IL-10 was shown in all off experiment groups compared to the control group. Conclusion : According to the results, we can expect that distilled red-ginseng Herbal Acupuncture may be further effccts in anti-cancer and immune improvement if increasing concentration.

• Asian-Australasian Journal of Animal Sciences
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• v.15 no.11
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• pp.1599-1605
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• 2002

In this study, three different experiments were conducted to evaluate the nutritional value of apple pomace produced in southern areas of the Republic of Korea (South Korea). The effects of combining apple pomace in different ratios with commercial concentrates and rice straw in the diets of Korean native goats (Capra hircus) were examined. In experiment I, in situ DM and CP disappearances from nylon bags incubated in the rumen of goats showed that greater amounts of DM and CP were released from apple pomace than those from concentrates at the later stages of incubation, but only after 48 h for DM and CP, respectively. This was reflected in the higher 'b' value of the slowly degradable fraction of the apple pomace compared to the concentrates. Prior to these times the trend was reversed. In experiment II, Korean native goats were fed a diet containing apple pomace with either rice straw or rice straw and concentrates, and the in vivo nutrient digestibilities compared to animals receiving an alfalfa hay. DM digestibility in the animals given apple pomace plus concentrates with rice straw (66.86%) were similar to the goats given alfalfa hay only (69.09%) but significantly greater than for a diet of rice straw plus concentrates. In experiment III, an in vivo study was conducted to investigate the inclusion of 30 to 60% apple pomace pre-mixed with rice straw, rice bran and concentrates on the nutritional value for Korean native goats. Apple pomace mixed diets had higher DM intakes, nutrient digestibility and nitrogen retention than diets without apple pomace, which may have been due to the higher non-structural carbohydrates (NSC) and less ADF and NDF than those in other treatments. Replacement of concentrates with apple pomace in rice straw based diets of Korean native goats fed either separately (experiment II) or by pre-mixing (experiment III) gave satisfactory feed intake, digestibility, pH of ruminal fluid and production of $NH_3$-N and VFA in the rumen of goats. The results of this study infer that apple pomace can be included at levels of up to 60% in the diets of goats without dramatic effect on the animal.

• Nuclear Engineering and Technology
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• v.56 no.5
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• pp.1942-1942
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• 2024

• Journal of the Korean Society of Food Science and Nutrition
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• v.29 no.1
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• pp.35-40
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• 2000

The effect of levanheptaose produced by levanase from Streptomyces sp. 366L on principle intestinal microflora was investigated. The reaction product, levanheptaose, was used as a carbon source for various intestinal microflora. As a results, Bifidobacterium adolescentis, Lactobacillus acidophilus, and Eubacterium limosum grew effectively in the in vitro experiment, whereas Clostridium perfringens, E. coli, and Staphylococcus aureus did not. Therefore levanheptaose seems to promote selectively the growth of B. adolescentis and L. acidophilus. In the in vivo experiment, the effect of levanheptaose on the growth of intestinal microflora, $\beta$-fructosidase activity, pH, and butyrate concentration were examined in rats. Apparently, the number of fecal Bifidobacteria, the amount of butyrate, and $\beta$-fructosidase activity were increased, whereas total aerobes and pH were reduced in rats fed by levanheptaose diets, compared with those of control diets. We concluded that those effects may be beneficial in improving gastrointestinal health.

• Journal of Animal Science and Technology
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• v.50 no.2
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• pp.199-208
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• 2008

This study was undertaken to investigate the effects of ruminally protected amino acids (Methionine and Lysine) on in vitro ruminal parameters, and in vivo milk yield and milk composition in mid-lactating cows. In the first in vitro experiment, there were no statistical significances between treatments in ruminal pH and dry matter digestibility during various incubation times. In the second in vivo experiment, milk yield decreased by 11.92% in control and 5.68% in the treatment respectively, but decrease rate of milk yield in the treatment was lower than control. Milk yields naturally decreased as time goes by since the DIMs(Days in milk) of the cows in experiment were in mid-lactation period. 4% FCM(Fat corrected milk) and milk protein yields also, respectively, decreased by 11.25% and 11.09% in control and 6.16% and 5.47% in the treatment as compared with the intial. Milk protein and milk fat production were higher in the treatment(0.90kg, 1.10kg) than those of control(0.66kg, 0.79kg). Milk fat content significantly increased with supplementing protected amino acids as compared to control(P<0.05). From the above results, protected amino acids were positively utilized in the performances of mid-lactating cows without inhibiting rumen fermentation. Further investigation is suggested for essential amino acid composition and intestinal digestion rate out of rumen bypass protein in dietary protein to be estimated.

• Environmental Mutagens and Carcinogens
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• v.16 no.1
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• pp.6-12
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• 1996

To investigate the clastogenicity of taxol and its precursor, 10-aleacetyl baccatin III, we performed chromosomal aberration assay with chinese hamster lung cells in vitro. The IC$_{50}$ values of taxol and 10-deacetyl baccatin III were determined as $1/16 \times 10^{-4}$ M (5.34 $\mu$g/ml) and $1 \times 10^{-2}$ M (560 $\mu$g/ml) in MTT assay, respectively. It means that the cytotoxicity of taxol revealed 100 times more cytotoxic than 10-deacetyl baccatin III in chinese hamster lung cell line. Nevertheless the strong positive genetic toxicity of taxol in the bone marrow micronucleus assay in vivo which was recently reported, we observed weak positive clastogenicity of taxoi only in the absence of metabolic activation system in the concentration ranges used in this experiment. Moreover, to clarify the involvement of metabolic fate of taxol because of its strong positive result in vivo, 10-deacetyl baccatin III which is a precursor in taxol synthesis, also subjected in chromosomal aberration assay in vitro. However, we observed no clastogenicity of 10-deacetyl baccatin III in this experiment. From above results, it was suggested that the esterification at C-13 appears to be relative for its genetic toxicity in chromosome aberration using chinese hamster lung cell in vitro.