• Asian-Australasian Journal of Animal Sciences
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• 제19권11호
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• pp.1574-1579
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• 2006

This study was performed to determine the developmental potentials of nuclear transfer (NT) embryos using life-span extended cells transfected with a foreign gene as donor cells. A life-span extended bovine embryonic fibroblast cell line was transfected with an expression vector in which the human type II collagen (BOMAR) and ear fibroblasts were used as a donor cell. Cytogenetic analysis was performed to analyze the chromosomal abnormality of donor cells. The fusion rate of 1.8 kV/cm for $15{\mu}sec$ given twice was significantly higher than that of other groups (p<0.05) and the embryos lysed were significantly higher after 1.8 kV/cm for $20{\mu}sec$ given once compared to other groups (p<0.01). The blastocyst development in the ear cell group was statistically significant compared to both BOMAR groups (p<0.01). Both BOMAR groups cultured more than 40 passages (>40 passages) had a lower number of chromosomes; however, fresh granulosa cell (GC) and BOMAR groups cultured less than 20 passages had normal chromosome numbers. Both >40 passages BOMAR groups had numerous obscure debris in metaphase spreads. The transfected foreign gene was expressed in all BOMAR groups, but not in the GC group. Based on these results, the lower developmental potential of NT embryos using life-span extended donor cells transfected with a foreign gene might be a cause of chromosomal abnormality in donor cells.

• 약학회지
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• 제38권3호
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• pp.311-321
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• 1994

Anticancer effects of Aloe on sarcoma 180 in ICR mouse or human cancer cells were determined. Sarcoma 180 cells were inoculated subcutaneously into male ICR mouse to determine effect of Aloe on tumor gowth, or inoculated intraperitoneally into male ICR mouse to determine effect of Aloe on life span prolongation, followed by oral administration of Aloe vera(10 mg/kg/day, 50 mg/kg/day) or Aloe arborescens(10 mg/kg/day, 100 mg/kg/day) once a day for 14 days. The administration of Aloe vera or Aloe arborescens did not suppress tumor growh. However the life span of ICR mouse was prolonged to 19%(p<0.05), 22%(p<0.05) and 32%(p<0.05) by administration of Aloe vera 10 mg/kg/day, Aloe vera 50 mg/kg/day, and Aloe arborescens 100 mg/kg/day, respectively. To determine anticancer effect of Aloe in vitro, Aloe extract was added to the culture of human gastric cancer cells(SNU-1) and colorectal cancer cells(SNU-C2A), and concentration of Aloe to inhibit cancer cell growth was determined using MTT(3-[ 4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) cytotoxicity assay. High $ID_{50}$ values of Aloe vera and Aloe arborescens against gastric cancer cell line(SNU-1) and colorectal cancer cell line(SNU-C2A) suggest that Aloe gel does not have anticancer effect on these specific human cancer cells although high concentration of Aloe inhibited growth of human cancer cells significantly.

• The Korean Journal of Physiology and Pharmacology
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• 제19권3호
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• pp.241-247
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• 2015

Ultraviolet (UV) radiation-induced loss of dermal extracellular matrix is associated with skin photoaging. Recent studies demonstrated that keratinocyte-releasable stratifin (SFN) plays a critical role in skin collagen metabolism by inducing matrix metalloproteinase 1 (MMP1) expression in target fibroblasts. In the present study, we examined whether SFN released from UVB-irradiated epidermal keratinocytes increases MMP1 release from dermal fibroblasts, and whether these events are affected by p-coumaric acid (p-CA), a natural phenolic compound with UVB-shielding and antioxidant properties. HaCaT cells were exposed to UVB in the absence and presence of p-CA, and the conditioned medium was used to stimulate fibroblasts in medium transfer experiments. The cells and media were analyzed to determine the expressions/releases of SFN and MMP1. UVB exposure increased SFN release from keratinocytes into the medium. The conditioned medium of UVB-irradiated keratinocytes increased MMP1 release from fibroblasts. The depletion of SFN using a siRNA rendered the conditioned medium of UVB-irradiated keratinocytes ineffective at stimulating fibroblasts to release MMP1. p-CA mitigated UVB-induced SFN expression in keratinocytes, and attenuated the MMP1 release by fibroblasts in medium transfer experiments. In conclusion, the present study demonstrated that the use of UV absorbers such as p-CA would reduce UV-induced SFN-centered signaling events involved in skin photoaging.

• 대한본초학회지
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• 제37권6호
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• pp.19-28
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• 2022

Objectives : To develop natural ingredients that help prevent or treat anti-inflammatory-related diseases and use themas basic data, we investigated anti-inflammatory activity of Cynanchi Atrati Radix Et Rhizoma water extracts(CWE) in lipopolysaccharide(LPS)-induced murine macrophage cell line, RAW 264.7 cells. Methods : The cell viabilities were evaluated with RAW 264.7 cells. The production of nitric oxide(NO), prostaglandin E2(PGE2), pro-inflammatory cytokines such tumor necrotic factor(TNF)-α and interleukin(IL)-6 were assessed in LPS-induced RAW 264.7 cell treated with CWE. Furthermore, the protein expression of inducible nitric oxide synthase (iNOS), cyclooxygenase-2(COX-2), and mitogen-activated protein kinase(MAPK) were assessed by western blotting. Results : In RAW 264.7 cell, the cell viability by CWE treatment was more than 98.4% at a concentration of 100-400 ㎍/mL. At a concentration of 800 ug/ml of CWE, the cell viability was as low as 86%. At doses of 100, 200 and 400 ㎍/mL, CWE inhibited the production of NO, PGE2, TNF-𝛼 and IL-6 in a dose-dependent manner and also decreased the expression of iNOS and COX-2 from LPS-induced RAW 264.7 cells. In addition, CWE significantly inhibited the MAPK pathway including decreased the phosphorylation of the p38, c-Jun N-terminal kinase(JNK) and extracellular signal-regulated kinase(ERK1/2). Conclusions : Our study provides evidence that CWE inhibits the production of main pro-inflammatory molecules in LPS-induced RAW 264.7 cells via expression of p38, JNK, and ERK1/2 MAPK signaling pathways. Therefore, CWE is expected to be widely used as a natural ingredient for anti-inflammatory functional foods or pharmaceuticals in the future.

• 한국축산식품학회지
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• 제43권4호
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• pp.563-579
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• 2023

Myogenesis and adipogenesis are the important processes determining the muscle growth and fat accumulation livestock, which ultimately affecting their meat quality. Hanwoo is a popular breed and its meat has been exported to other countries. The objective of this study was to compare the myogenesis and adipogenesis properties in satellite cells, and meat quality between Hanwoo and Vietnamese yellow cattle (VYC). Same 28-months old Hanwoo (body weight: 728±45 kg) and VYC (body weight: 285±36 kg) steers (n=10 per breed) were used. Immediately after slaughter, tissue samples were collected from longissimus lumborum (LL) muscles for satellite cells isolation and assays. After 24 h post-mortem, LL muscles from left carcass sides were collected for meat quality analysis. Under the same in vitro culture condition, the proliferation rate was higher in Hanwoo compared to VYC (p<0.05). Fusion index was almost 3 times greater in Hanwoo (42.17%), compared with VYC (14.93%; p<0.05). The expressions of myogenesis (myogenic factor 5, myogenic differentiation 1, myogenin, and myogenic factor 6)- and adipogenesis (peroxisome proliferator-activated receptor gamma)-regulating genes, and triglyceride content were higher in Hanwoo, compared with VYC (p<0.05). Hanwoo beef had a higher intramuscular fat and total monounsaturated fatty acids contents than VYC beef (p<0.05). Whilst, VYC meat had a higher CIE a* and total polyunsaturated fatty acids content (p<0.05). Overall, there was a significant difference in the in vitro culture characteristics and genes expression of satellite cells, and meat quality between the Hanwoo and VYC.

• 생명과학회지
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• 제19권12호
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• pp.1690-1697
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• 2009

CD11c와 CD80 및 CD86과 같은 보조 수용체는 주로 수지상 세포에서 발현되는 세포 표지 인자이다. 본 연구에서는 KG-1, HL-60, NB4 및 THP-1 세포와 같은 여러 종류의 백혈병 세포를 이용하여 이들 세포에 재조합 Flt-3 리간드를 처리하였을 때 수지상 세포의 표면 인자인 CD11c의 발현에 어떠한 변화가 있는가를 조사하였다. KG-1 세포뿐만 아니라 NB4세포와 HL-60 세포에서도 Flt-3 수용체가 발현됨을 확인하였으나 THP-1 세포에서는 이들 수용체가 발현되지 않았다. KG-1 세포를 Flt-3 리간드나 granulocyte macrophage-colony stimulating factor (GM-CSF)와 tumor necrosis factor (TNF)-$\alpha$를 섞은 배양액에서 배양하였을 때 세포 증식은 억제되었으며 CD11c 발현은 현저히 증가되었다. 그러나 Flt-3 리간드를 처리한 KG-1세포에서는 GM-CSF와 TNF-$\alpha$를 처리한 세포에서와는 다르게 major histocompatibility complex (MHC)-I 및 MHC-II의 발현은 증가되지 않았다. Flt-3 리간드는 HL-60 세포와 NB4 세포의 CD11c 발현도 증가시켰으나 THP-1 세포에서는 아무런 영향이 없었다. CD11c의 발현과 비교하여 CD11b의 발현은 Flt-3 리간드에 의하여 KG-1 세포에서는 약하게 증가하였으나 NB4 세포와 HL-60 세포에서는 증가되지 않았다. KG-1 세포를 Flt-3 리간드로 처리하였을 때 extracellular signal-regulated kinase-1/2 (ERK-1/2)와 p38-mitogen-activated protein kinase (p38-MAPK)의 단백질 인산화가 증가되었으며 Flt-3 리간드에 의한 CD11c 발현의 증가는 MEK의 억제제인PD98059에 의하여 사라짐을 확인하였다. 본 연구 결과는 Flt-3 수용체 리간드의 처리에 의하여 $CD34^+$ myelomonocyte분화 단계인 KG-1 세포와 promyelocyte 분화 단계의 백혈병 세포에서 수지상 세포와 유사한 세포 형으로 분화된다는 것을 보였고 Flt-3 수용체 리간드에 의한 이들 백혈병 세포의 수지상 세포유사 세포로의 분화는 ERK-1/2의 활성화에 의하여 일어날 수 있음을 보여 준다.

• Animal cells and systems
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• 제9권1호
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• pp.19-25
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• 2005

This study was conducted to determine the effect of dietary salt on the synthesis of glucocorticoids in the adrenal cortex of mice. Mice had ad libitum access to 3% sodium chloride as the only drinking fluid (high salt diet) for either 4 days or 4 weeks. Adrenocortical expression of cytochrome P450 11beta-hydroxylase, a major regulatory enzyme in the biosynthesis of glucocorticoids, was examined by immunohistochemistry and western blot analysis. Ultrastructure of adrenocortical cell and plasma level of corticosterone were analyzed as well. Size and density of lipid droplets in the cortical cell were increased by high salt diet. Four days of high salt diet decreased P450 11beta-hydroxylase in the adrenal cortex, but 4 weeks increased it. Plasma level of corticosterone changed in parallel with the Cortical level of P450 11 beta-hydroxylase. These results suggest that high salt diet may modulate the biosynthesis of glucocorticoids, at least partly, via regulating the expression of P450 11beta-hydroxylase in adrenocortical cells.

• 환경생물
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• 제23권3호
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• pp.221-227
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• 2005

The effects of light and $CO_2$ on the electrophysiological characteristics of guard cells in the intact leaf and isolated epidermis have been investigated. Fast hyperpolarization of guard cell apoplastic PD in the intact leaf was recorded reaching up to around 7 mV and 20 mV in response to light and $CO_2$. Whenever the experiments were attempted with isolated epidermis, there was no response to light and $CO_2$. In order to determine the influence of the mesophyll cells, the apoplastic PD of guard cells in isolated epidermis was measured in the presence of the mesophyll supernatant or the control medium. The apoplastic PD in isolated epidermis was hyperpolarized to -7mV, changing from -22mV to -29mV at 40 min. But, when isolated epidermis was incubated with the supernatant from mesophyll cells incubated in the light, the apoplastic PD in isolated epidermis was hyperpolarized to -19 mV, changing from -22 mV to -40.5 mV. $CO_2$ also caused a change of 0.1 to 0.3 pH unit in the intact leaf. However, this change was absent in isolated epidermis. A vibrating probe was used to detect the change in electrical currents at the surface of excised intact leaves and isolated epidermis. The reading of excised intact leaves in the dark was $0.5\muA\;cm^{-2},$ remaining steady until illuminated. Light increased the current on the surface of excised leaves to about $0.8\muA\;cm^{-2},$. However, light had no effect in the current on the surface of isolated epidermis. Apoplastic pH changes across the stomatal complex in response to light and dark were measured both in the intact leaves and isolated epidermis over the same time period using pH micro-electrodes. The guard cell wall of intact leaf was acidified to 2.5 pH unit, falling from pH 7.5 to pH 5.0 in the first 10 min. in the light. At the same time the guard cell wall pH of isolated epidermis fell from pH 7.5 to pH 7.0 at 10 min. The guard cell wall pH of isolated epidermis incubated in the mesophyll supernatant fell from pH 7.6 to pH 6.7 at 10 min. Likewise, It could be imagined that an electrical signal, chemicals and hormones propagated from the mesophyll in response to light and $CO_2$ could control a fast stomatal response.

• Journal of Microbiology and Biotechnology
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• 제19권10호
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• pp.1103-1108
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• 2009

The human immunodeficiency virus (HIV)-l protein Tat acts as a transcription transactivator that stimulates expression of the infected viral genome. It is released from infected cells and can similarly affect neighboring cells. The nucleocapsid is an important protein that has a related significant role in early mRNA expression, and which contributes to the rapid viral replication that occurs during HIV-1 infection. To investigate the interaction between the Tat and nucleocapsid proteins, we utilized cDNA micro arrays using pTat and flag NC cotransfection in HEK 293T cells and reverse transcription-polymerase chain reaction to validate the micro array data. Four upregulated genes and nine downregulated genes were selected as candidate genes. Gene ontology analysis was conducted to define the biological process of the input genes. A proteomic approach using PathwayStudio determined the relationship between Tat and nucleocapsid; two automatically built pathways represented the interactions between the upregulated and downregulated genes. The results indicate that the up- and downregulated genes regulate HIV-1 replication and proliferation, and viral entry.

• 한국미생물·생명공학회지
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• 제19권1호
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• pp.70-75
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• 1991

Continuous and simultaneous production of gluconic acid and sorbitol from glucose and fructose was carried out by using glucose-fructose oxidoreductase and glucanolactonase of Zymomonas mobilis. In order to utilize the enzymes without purification, Zymomonas mobilis was permeabilized with toluene. Optimum conditions for permeabilization and reaction kinetics of permeabilized Zymomonas mobilis were studied. In batch operation with the permeabilized cells immobilized in alginate beads, about 90% conversion was obtained within 35 h reaction. Continuous production of gluconic acid and sorbitol using the immobilized permeabilized cells was carried out. Optimum conditions for continuous operation with the imn~obilized cells were; pH 6.2 and temperature $40^{\circ}C$. Maximum productivities for gluconic acid and sorbitol were about 14.5 g/l/h and 14.8 g/l/h respectively at the dilution rate of 0.075 $h^{-1}$ when 300 g/l each of substrates was fed.