• Journal of Animal Science and Technology
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• v.58 no.9
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• pp.33.1-33.7
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• 2016

Background: The continuing growth of the ethanol industry has generated large amounts of various distillers grains co-products. These are characterized by a wide variation in chemical composition and ruminal degradability. Therefore, their precise formulation in the ruminant diet requires the systematic evaluation of their degradation profiles in the rumen. Methods: Three distillers grains plus soluble co-products (DDGS) namely, corn DDGS, high-protein corn DDGS (HP-DDGS), and wheat DDGS, were subjected to an in situ trial to determine the degradation kinetics of the dry matter (DM) and crude protein (CP). Soybean meal (SBM), a feed with highly degradable protein in the rumen, was included as the fourth feed. The four feeds were incubated in duplicate at each time point in the rumen of three ruminally cannulated Hanwoo cattle for 1, 2, 4, 6, 8, 12, 24, and 48 h. Results: Wheat DDGS had the highest filterable and soluble A fraction of its DM (37.2 %), but the lowest degradable B (49.5 %; P < 0.001) and an undegradable C fraction (13.3 %; P < 0.001). The filterable and soluble A fraction of CP was greatest with wheat DDGS, intermediate with corn DDGS, and lowest with HP-DDGS and SBM; however, the undegradable C fraction of CP was the greatest with HP-DDGS (41.2 %), intermediate with corn DDGS (2.7 %), and lowest with wheat DDGS and SMB (average 4.3 %). The degradation rate of degradable B fraction ($%\;h^{-1}$) was ranked from highest to lowest as follows for 1) DM: SBM (13.3), wheat DDGS (9.1), and corn DDGS and HP-DDGS (average 5.2); 2) CP: SBM (17.6), wheat DDGS (11.6), and corn DDGS and HP-DDGS (average 4.4). The in situ effective degradability of CP, assuming a passage rate of $0.06h^{-1}$, was the highest (P < 0.001) for SBM (73.9 %) and wheat DDGS (71.2 %), intermediate for corn DDGS (42.5 %), and the lowest for HP-DDGS (28.6 %), which suggests that corn DDGS and HP-DDGS are a good source of undegraded intake protein for ruminants. Conclusions: This study provided a comparative estimate of ruminal DM and CP degradation characteristics for three DDGS co-products and SBM, which might be useful for their inclusion in the diet according to the ruminally undegraded to degraded intake protein ratio.

• Journal of dental hygiene science
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• v.15 no.2
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• pp.209-219
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• 2015

In order to explore an effect of interaction of Streptococcus gordonii, Fusobacterium nucleatum and Porphyromonas gingivalis that are bacteria relevant to periodontal disease on its growth, the bacteria were incubated in trypticase soy hemin menadione broth at $37^{\circ}C$ $CO_2$ incubator for 7 days through anaerobic jar by single and co-culture with heat treated dead bacteria under anaerobic gas pack. In order to confirm growth level, absorbance was measured and for confirming colony structure and form, it was observed with scanning electron microscope. In order to confirm an effect on pathogenicity of P. gingivalis, real time reverse transcriptase polymerase chain reaction was implemented for expression analysis for rgpA gene that produces HRgpA which is gingipain. As a result, the following conclusion was obtained. Colony formation of S. gordonii and P. gingivalis was increased by other dead bacteria and in case of F. nucleatum, its colony formation was showed an aspect of being increased by dead bacterium of P. gingivalis but decreased by dead bacterium of S. gordonii. Therefore, it is considered that the strains being used for this study would affect interactively through bacterial cell itself as well as their interaction factor at the time of colony formation.

• Journal of Periodontal and Implant Science
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• v.23 no.1
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• pp.77-96
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• 1993

This in vitro study was undertaken to observe whether citric acid application aids the attachment and proliferation of human periodontal ligament cells to the root surfaces of periodontally diseased teeth. The roots were prepared so that the comparison could be made among the control healthy root surface, citric acid demineralized and non-demineralized root planted surfaces. Prior to the cell attachment experiment, each groups were prepared for scanning electron microscopic (SEM) examinations of root surface morphology, All specimens were fixed with phosphate buffered glutaraldehydes, postfixed with phosphate buffered osmium tetraoxide and stained with phosphate buffered tannic acid. dehydrated in ethanol, critical point dried, sputter coated with gold and examined under the SEM. In the cell attachement experiment, human cultured periodontal ligament cells at concentration to $4.5{\times}\;10^4\;cells/ml$ were seeded in each culture well which contained prepared roots and incubated for 30min 1, 2, 6, 12 and 24 hours at 37, 5% $CO_2$air incubator. Than the specimens were prepared for SEM examination using, the same methods as described above. In the cell proliferation experiment, $5{\times}\;10^4\;cells/ml$ cells were seeded incubated with the specimens for 6 hours. Then, all of the specimens were moved into fresh culture well and incubated for 24, 48, and 72 hours. The cell counting was done after trypsinization, under light microscope. The results were as follows. When viewed the surface morphology prior to the cell attachment, the non acid treated root planed surface displayed scaling striation and occasional bacteria and calculus. The citric acid treated specimens displayed little debris on the surface and funnel shaped orifices of dentinal tubules. There were no apparent differences in the morphology of cells attached to the control and experiment groups. However, in initial attachement, there was a slight more enhanced appearance in attachment in citric acid treated groups than other root surfaces. After 6 hours of incubation, most of the cells initiated the alteration of cell morphology from ovoid to spindle shapes. After 24 hours of incubation, most of the cells displayed proliferated appearance and connected with each other via numerous processes. In the cell proliferation experiments, there were statistically significant increased number of cells in citic acid treated groups than other groups.

• Journal of Periodontal and Implant Science
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• v.25 no.3
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• pp.587-602
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• 1995

The purpose of this study was to compare the effects of citric acid and tetracycline HCI application to the root surfaces of periodontally diseased teeth on the proliferation and spreading of human periodontal ligament cells. The roots were prepared so that the comparison could be made among root planed, citric acid treated and tetracycline HCI treated surfaces. In the cell proliferation experiment, human periodontal ligament cells at a concentration of $1{\times}10^5$ cells/ml were seeded in each culture well with specimens and incubated for 6 hours. Then, the specimens were transferred to a fresh culture well and incubated for 24, 48, 72 hours respectively. The cell counting was done after trypsinization. In the cell spreading experiment, $1{\times}10^4$ cells/ml were seeded in each culture well and incubated for 30min, 6 hours and 24 hours at 37.5$^{\circ}C$ in a $CO_2$ incubator. Then, all specimens were fixed with phosphate buffered glutaraldehydes, postfixed with phosphate buffered osmium tetraoxide, stained with phosphate buffered tannic acid, dehydrated in ethanol, dried at a critical point, coated with gold and examined under a scanning electron microscope. The results were as follows:In the cell proliferation experiments, the number of attached cells increased more in the tetracycline treated group than in the other groups. In the initial attachment, the appearance of the tetracycline treated the groups was slightly more spread out than in the other groups. After 6 hours of incubation, it was observed in most of the cells that cell morphologic alteration went from ovoid shapes sto spindle shapes. After 24 hours of incubation, the cells of all groups had a fusiform appearance and were connected to each other by numerous cytoplasmic processes. The tetracycline and citric acid treated groups had a similar spreading appearance of periodontal ligament cells, but the tetracycline treated group was more effective in the cell proliferation than the citric acid group.

• The korean journal of orthodontics
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• v.27 no.3 s.62
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• pp.503-513
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• 1997

This study was undertaken to investigate the effect of oxygen tension on the activity and function of the cells derived from human periodontal ligament by measuring cell activity, total protein synthesis, collagen synthesis, $IL-1{\beta},\;IL-6,\;TNF-{\alpha}$ Human periodontal ligament fibroblasts were collected from premolars extracted for orthodontic treatment and incubated in the environment of $37^{\circ}C,\;5\%\;CO_2,\100\%$ humidity. After the fifth to sixth passage they were used for the experiment. Gaspack system to which $0.2{\mu}m$ Millipore filter was attached was connected to mixed-gas tanks. The mixed gases were composed of $10\%\;O_2,\;5\%\;CO_2,\;85\%\;N_2$ in hyoxic group or $90\%\;O_2,\;5\%\;CO_2,\;5\%\;N_2$ in hyperoxic group and $5\%\;CO_2,\;95\%$ air for control. After incubation in $37^{\circ}C$ for 2, 4, 6 days, cell activity was determined by tetrazolium(MTT) assay and total protein synthesis was assayed using sulforhodamine B(SRB). And measurement of 4-hydroxyproline was performed to assess collagen synthesis md $IL-1{\beta},\;IL-6,\;and\;TNF-{\alpha}$ were measured by enzymeimmunoassay. The results were as follows. 1. The cell activity and total protein synthesis in hypoxic group were a little higher than or almost the same with those in control group. 2. In hyperoxic group, the cell activity was lower than that in control group and total protein synthesis was decreased. 3. Collagen synthesis was significantly decreased initially in both hypoxic and hyperoxic group and increased nearly to the level of control group as the duration of cell incubation was longer 4. As a result of enzymeimmunoassay, the amount of cytokines was $IL-6,\;TNF-{\alpha}\;and\;IL-1{\beta}$ in order. 5. $IL-6,\;TNF-{\alpha}\;and\;IL-1{\beta}$ were increased more rapidly in both hypoxic and hyperoxic group than in control group as the duration of cell incubation was longer. 6. There were more $IL-6\;and\;TNF-{\alpha}$ in hyperoxic group than in control group after 6 days, and there were more $IL-6\;and\;TNF-{\alpha}$ after 6 days than after 2 or 4 days in hyperoxic group. These results suggested that oxygen tension might modulate the production of extracellular matrix and cytokines in the cells derived from human periodontal ligament.

• Archives of Pharmacal Research
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• v.25 no.6
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• pp.903-909
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• 2002

Research in the cytokine field has grown exponentially in recent years, and the validity of such studies relies heavily on the appropriate measurement of levels of cytokines in various biological samples. Transforming growth factor (TGF)-$\beta$, a hormonally active polypeptide found in normal and transformed tissue, is a potent regulator of cell growth and differentiation. The most widely used bioassay for TGF-$\beta$ is the inhibition of the proliferation of mink lung epithelial cells. Though detection of [$^3$H]thymidine incorporation is more sensitive than the MTT assay, it presents some disadvantages due to the safety and disposal problems associated with radioisotopes. In this study, we attempted to ascertain the experimental conditions which could be used for measuring the in vitro biological activity of TGF-$\beta$ in a safer and more sensitive way compared with the currently available methods. We compared the commonly used method, the MTT assay, to the XTT assay using different parameters including cell number, incubation time and the wave length used for detecting the product. We examined the anti-proliferative activities of TGF-$\beta$ in three different cell lines: Mv-1-Lu mink lung epithelial cells, MCF10A human breast epithelial cells and H-ras-transformed MCF10A cells. Herein, we present an experimental protocol which provides the most sensitive method of quantifying the biological activity of TGF-$\beta$, with a detection limit of as low as 10 pg/ml: Mv-1-Lu or H-ras MCF10A cells ($1{\times}10^5/well$) were incubated with TGF-$\beta$ at $37^{\circ}C$ in a humidified $CO_2$ incubator for 24 hr followed by XTT treatment and determination of absorbance at 450 or 490 nm. Our results may contribute to the establishment of an in vitro bioassay system, which could be used for the satisfactory quantitation of TGF-$\beta$.

• Korean Journal of Medicinal Crop Science
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• v.22 no.3
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• pp.223-230
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• 2014

This study was performed to enhance contents of low molecular ginsenoside using steaming and fermentation process in low quality fresh ginseng. For increase in contents of Rg2, Rg3, Rh2 and CK in low quality fresh ginseng, a steaming process was applied at $90^{\circ}C$ for 12 hr which was followed by fermentation process at Lactobacillus rhamnosus HK-9 incubated at $36^{\circ}C$ for 72 h. The contents of ginsenoside Rg1, Rb1, Rc, Re and Rd were decreased with the steaming associated with fermentation process but ginsenoside Rg2, Rg3, Rh2 and CK increased after process. It was found that under the steaming associated with fermentation process, low molecule ginsenosides such as Rg2, Rg3, Rh2 and CK were increased as 3.231 mg/g, 2.585 mg/g and 1.955 m/g and 2.478 mg/g, respectively. In addition, concentration of benzo[${\alpha}$]pyrene in extracts of the low quality fresh ginseng treated by the complex process was 0.11 ppm but it was 0.22 ppm when it was treated with the steaming process. This result could be caused by that the most efficiently breakdown of 1,2-glucoside and 1,4-glucoside linkage to backbone of ginsenosides by steaming associated with fermentation process. This results indicate that steaming process and fermenration process can increase in contents of Rg2, Rg3, Rh2 and CK in low quality fresh ginseng.

• Asian-Australasian Journal of Animal Sciences
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• v.18 no.2
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• pp.158-164
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• 2005

A practical approach was proposed to produce transgenic chimeric chickens using blastodermal cells (BCs). The chicken BCs were mechanically dissociated and transferred into the recipient eggs that had been exposed to 500 rads irradiation of$^{60}Co$ and windowed on the equatorial plane. Chimeric chickens were generated using two models: the crosses (MXL) from Black Minors (ii,EE,b/b) ♂${\times}$Barred Leghorns (ii,ee,B/-) ♀ as donors and White Leghorns (WL, II) as acceptors (Model 1), or the Black Heifengs (BH, ii,EE,bb) as donors and Hua-xing white (HW, II) as recipients (Model 2). The treated eggs were incubated in their original shells in normal conditions until hatching. Green fluorescent protein (GFP) gene was transferred into the BCs derived from MXL and BH via lipofectamine and the pEGFP-C1, and transfection efficiency into the BCs was examined under a fluorescent microscope. Potential transgenic chimeras were selected based on the proposed methods in this study. Using the fresh BCs, the best rate of phenotypic chimeras was 6.7% and 26.0% in model-1 groups, and model-2 groups, respectively. We also described the optimized conditions for transfection. Although 30% of the BCs transfected in vitro emitted green light under an inverted fluorescent microscope, no embryos injected with the transfected BCs expressed foreign GFP gene at 3-4 days.

• The Journal of Internal Korean Medicine
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• v.29 no.4
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• pp.846-855
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• 2008

Objects : The aim of this study was to investigate the influence of five herbal medicines on cytochrome P450 3A4 drug-metabolizing enzymes in human liver microsomes. Methods : To use human liver microsomes, an extract of five herbal medicines, which are Artemisia princeps Pampan, Sophora jeponica Linne, Panax notoginseng F. H. Chen, Lithospermum Erythrorhizon Sieb., and Cirsium maackii Maxim, which together are called Jihyulyak(止血藥, drugs for arresting bleeding, hemostatics), was co-incubated and measured for relative enzyme activity in incubation condition compared to ketoconazole, a representative inhibitor of CYP 3A4. Results : We showed that all five of the traditional herbal medicines had no inhibition effect of CYP 3A4 at 10, 20, 30, 40, and $50{\mu}g/ml$ doses in human liver microsomes, although Sophora japonica Linne(SJL) showed a little inhibition at about 81% inhibition rate of control. However, this result is not enough to prove that SJL has a CYP 3A4 inhibition effect. Moreover, we can't make sure that those rates had significant induction effect on CYP 3A4. Conclusions : The result of this study could support that those herbal medicines are safer than chemical drugs, even if this is the basic step to prove that result.

• Journal of Ecology and Environment
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• v.35 no.1
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• pp.1-7
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• 2012

To calculate and predict soil carbon budget and cycle, it is important to understand the complex interrelationships involved in soil respiration rate (Rs). We attempted to reveal relationships between Rs and key environmental factors, such as soil temperature, using a laboratory incubation method. Soil samples were collected from mature deciduous (MD), mature coniferous (MC), immature deciduous (ID), and immature coniferous (IC) forests. Prior to measure, soils were pre-incubated for 3 days at $25^{\circ}C$ and 60% of maximum water holding capacity (WHC). Samples of gasses were collected with 0, 2, and 4 h interval after the beginning of the measurement at soil temperatures of 5, 15, 25, and $35^{\circ}C$ (at 60% WHC). Air samples were collected using a syringe attached to the cap of closed bottles that contained the soil samples. The $CO_2$ concentration of each gas sample was measured by gas chromatography. Rs was strongly correlated with soil temperature (r, 0.93 to 0.96; P < 0.001). For MD, MC, ID, and IC soils taken from 0-5 cm below the surface, exponential functions explained 90%, 82%, 92%, and 86% of the respective data plots. The temperature and Rs data for soil taken from 5-10 cm beneath the surface at MD, MC, ID, and IC sites also closely fit exponential functions, with 83%, 95%, 87%, and 89% of the data points, respectively, fitting an exponential curve. The soil organic content in mature forests was significantly higher than in soils from immature forests (P < 0.001 at 0-5 cm and P < 0.005 at 5-10 cm) and surface layer (P = 0.04 at 0-5 cm and P = 0.12). High soil organic matter content is clearly associated with high Rs, especially in the surface layer. We determined that the incubation method used in this study have the possibility for comprehending complex characteristic of Rs.