• Journal of Life Science
• /
• v.7 no.2
• /
• pp.79-87
• /
• 1997

To determine the cause of Vibrio septicemia by understanding the characteristics endotoxin from Vibrio vulnificus, lethal dose, heat resistance and vascular permeability enhancing activity were svaluated using vegestative cell and cell homogenate and the result is as follows: 1. Vibrio vulnificus CDC B3547 of patient origin did not exihibit any significant difference in toxicity compared to Vibrio vulnificus B57 of enviroment origin. 2. Strong toxicity was observed when viable cell count of Vibrio vulnificus CDC B3547 was more than 10$^{7}$/ml. 3. Toxicity of cell homogenate was completely inactivited upon geating at 80$^{\circ}$C for 20min. 4. Cell homogenate did not show hemolyic activity but was acknowleged to have cytotoxicity. 5. Major lethal toxin against mouse was existed in Vibrio vulnificus CDC B3547; however, separation of LPS and LPS-protein complex was not successful using the current technique.

• 한국생물공학회:학술대회논문집
• /
• 2005.04a
• /
• pp.515-515
• /
• 2005

• Journal of Physiology & Pathology in Korean Medicine
• /
• v.19 no.6
• /
• pp.1666-1672
• /
• 2005

Oriental medicinal drugs have a broad spectrum of clinical use for the cure of nervous system diseases including brain ischemic damages or neuropathies. Yet, specific drugs or drug components used in the oriental medicine in relation to none fiber regeneration are not known. In the present study, possible growth promoting effects of oriental medicinal drugs were investigated in the injured sciatic nerve system in the rat. By immunofluorescence staining, we found that Jahageo (JHG, Hominis placenta) increased Induction levels of axonal growth associated protein GAP-43 in the rat sciatic none. Small growth promoting activity was found in Golsebo (GSB, Drynariae rhizoma) and Baikhasuo (BHSO, Polygoni multiflori radix) drugs. JHG also increased cell cycle protein Cdc2 levels in the injured area of the sciatic nerves. Immunofluorescence staining indicated that induced Cdc2 protein was mostly localized in the Schwann cells in the injury area, implying that JHG activity might be related to increased Schwann cell proliferation during axonal regeneration. Moreover, levels of phospho-extracellular signal-regulated (ERK) pathway in the injured neNes were elevated by JHG treatment while levels of total ERK were unaltered. In vivo measurement of axonal regeneration using retrograde tracer showed that JHG, GSB and BHSO significantly enhanced Dil-labeled regenerating motor neurons compared with saline control. The present data suggest that oriental medicinal drugs such as JHG, GSB, and BHSO may be a useful target for developing specific drugs of axonal regeneration.

• The Journal of Korean Medicine
• /
• v.29 no.5
• /
• pp.14-28
• /
• 2008

Objective : This study was carried out to understand effects of ginseng(hearinafter ; GS, Panax Ginseng) extract on regeneration responses on injured sciatic nerves in rats. Methods :Using white mouse, we damaged sciatic nerve & central nerve, and then applied GS to the lesion. Then we observed regeneration of axon and non-neuron. Results : 1. NF-200 protein immunostaining for the visualization of axons showed more distal elongation of sciatic nerve axons in GS-treated group than saline-treated control 3 and 7 days after crush injury. 2. GAP-43 protein was increased in the injured sciatic nerve and further increased by GS treatment. Enhanced GAP-43 protein signals were also observed in DRG prepared from the rats given nerve injury and GS treatment. 3. GS treatment in vivo induced enhanced neurite outgrowth in preconditioned DRG sensory neurons. In vitro treatment of GS on sensory neurons from intact DRG also caused increased neurite outgrowth. 4. Phospho-Erk1/2 protein levels were higher in the injured nerve treated with GS than saline. Phospho-Erk1/2 protein signals were mostly found in the axons in the injured nerve. 5. NGF and Cdc2 protein levels showed slight increases in the injured nerves of GS-treated group compared to saline-treated group. 6. The number of Schwann cell population was significantly increased by GS treatment in the injured sciatic nerve. GS treatment with cultured Schwann cells increased proliferation and Cdc2 protein signals. 7. GS pretreatment into the injured spinal cord generated increased astrocyte proliferation and oligodendrocytes in culture. In vitro treatment of GS resulted in more differentiated pericytoplasmic processes compared with saline treatment. 8. More arborization around the injury cavity and the occurrence at the caudal region of CST axons were observed in GS-treated group than in saline-treated group. Conclusion :GS extract may have the growth-promoting activity on regenerating axons in both peripheral and central nervous systems.

• Microbiology and Biotechnology Letters
• /
• v.42 no.1
• /
• pp.73-81
• /
• 2014

Salsola collina, also known as Russian thistle, is widely distributed in and around waste facilities, roadsides, and drought and semi-drought areas, and is used as a traditional folk remedy in Chinese medicine for the treatment of hypertension. In this study, we have evaluated the anti-oxidative and anti-cancer activities of the ethanol extract of S. collina Pall. (EESC), and the molecular mechanisms of its anti-cancer effects on human colon carcinoma HT29 cells. EESC exhibited anti-oxidative activity through DPPH radical scavenging capacity and showed cytotoxic activity in a dose-dependent manner in HT29 cells. After EESC treatment, HT29 cells altered their morphology, becoming smaller and irregular in shape. EESC also induced cell accumulation in the G2/M phase in a dose-dependent manner, accompanied by a decrease of cell population in the G1 phase. The G2/M arrest by EESC was associated with the increased expression of cyclin-dependent kinase (CDK) inhibitor p21 and Wee1 kinase, which phosphorylates, or inactivates, Cdc2. EESC treatment induced the phosphorylation of Cdc2 and Cdc25C, and inhibited cyclin A and Cdc25C protein expression. In addition, S arrest was induced by the highest concentration of EESC treatment, associated with a decrease of cyclin A and Cdk2 expression. These findings suggest that EESC may possess remarkable anti-oxidative activity and exert an anti-cancer effect in HT29 cells by cell cycle regulation.

• Proceedings of the Korean Society of Toxicology Conference
• /
• 2006.11a
• /
• pp.46-54
• /
• 2006

The methylmercury (MeHg) is a toxic environmental pollutant, causing serious neurological and developmental effects in humans. Recent epidemiological studies have indicated that ingestion of MeHg in fish during pregnancy can result in neuroethological effects in the offspring. However, the mechanism underlying the MeHg-toxicity is not fully understood. To elucidate the mechanisms of toxicity of MeHg and of defense against MeHg, we searched for factors that determine the sensitivity of yeast cells to MeHg, and found that overexpression of Cdc34, a ubiquitin-conjugating enzyme (E2) that is a component of the ubiquitin-proteasome (UP) system, induces a resistance to MeHg toxicity in both yeast and human cells. The UP system is involved in the intracellular degradation of proteins. When Cdc34 is overexpressed in cells, ubiquitination reactions are activated and the degradation of certain proteins by the UP system is enhanced. Therefore, it seems likely that certain as-yet-unidentified proteins that increase MeHg toxicity might exist in cons and that toxicity might be reduced by the enhanced degradation of such proteins, mediated by the UP system, when Cdc34 is overexpressed. SCF ubiquitin-ligase is a component of UP system and consists of Skpl, the scaffold protein Cdc53, the RING-finger protein Hrt1, and one member of the family of F-box proteins. The F-box proteins directly bind to the substrates and are the determinants of substrate specificity of SCF. Therefore, we searched for the f-box protein that cofers resistance to MeHg, and found that overexpression of Hrt3 or Yi1224w induced resistance to MeHg toxicity in yeast cells. Since the protein(5) that enhance toxicity of MeHg might plausibly be induced in substrates of both f-box proteins, we next searched for substrate proteins that are recognized by Hrt3 or Y1r224w using two-hybrid screen. We found that Did3 or Crsl interacts with Hrt3; and Eno2 interacts with Yir224w. The yeast cells that overexpressed each those proteins showed hypersensitivity to MeHg, respectively, indicating that those proteins enhance the MeHg toxicity. Both Dld3 and Eno2 are proteins involved in the synthesis of pyruvate, and overexpression of both proteins might induce increase in interacellular levels of pyruvate. Deletion of Yi1006w that transports pyruvate into the mitochondria induced aresistance to MeHg. These results suggest that the promotion of the pyruvate irdlowinto the mitochondria might enhance MeHg toxicity. This study providesimportant keyfor the elucidauon of the molecular mechanism of MeHg toxicity.

• The Journal of Korean Medicine
• /
• v.31 no.1
• /
• pp.93-111
• /
• 2010

Objectives: The present study was performed to evaluate the potential effects of Bupleuri radix (SH) on regenerative activities in the peripheral sciatic nerve after crushing injury in rats. Methods: Axonal regeneration after crush injury in rats was analyzed by immunofluorescence staining using anti-NF-200 antibody and retrograde tracing of DiI-axons. Changes in protein levels in the sciatic nerve axons and DRG tissue were analyzed by Western blot analysis and immunofluorescence staining. Effects of SH extract treatment on neurite outgrowth was examined by immunofluorescence staining for cultured DRG neurons. Results: Major findings on the effects of SH extract treatment on axonal regeneration are summarized as follows. 1. SH-mediated enhancement in axonal regeneration was identified by immuno- fluorescence straining of NF-200 protein and retrograde tracing of DiI-labeled axons. 2. Axonal GAP-43 protein levels were upregulated by SH not only in the injured axons but also in the DRG sensory neurons corresponding to sciatic sensory axons. 3. Phospho-Erk1/2 protein levels were increased in both injured axonal area and DRG sensory neurons by SH. Phospho-Erk1/2 was also found in non-neuronal cells in the injured axons. 4. SH elevated levels of Cdc2 protein produced in Schwann cells in the distal portions of injured sciatic nerves. 5. The neurite outgrowth of DRG sensory neurons in culture was augmented by SH, and these changes were positively associated with GAP-43 production levels in the DRG neurons. Conclusions: These data suggest that SH extract improves the regenerative responses of injured peripheral neurons, and thus may be useful for understanding molecular basis for the development of therapeutic strategies.

• The Microorganisms and Industry
• /
• v.19 no.2
• /
• pp.2-10
• /
• 1993

이 총론에서는 cell cycle의 조절에 관계하는 p34cdc kinase의 특성과 기질 그리고 cell cycle에서의 역할을 살펴보고, 또 cell cycle에서와 여러가지 세포내의 현상에 중요한 역할을 하는 것으로 알려진 casein kinase II의 특성과 기질 그리고 cell cycle에서의 역할을 살펴보고자 한다. 그리고 이런 효소들을 연구하는 데 필수적인 방법이나 시약들로 소개하고자 한다.

• Journal of Physiology & Pathology in Korean Medicine
• /
• v.24 no.6
• /
• pp.1042-1052
• /
• 2010

This study aimed to evaluate the protective effects of Mundongcheongpye-eum (MCE) on elastase-induced lung injury. The extract of MCE was treated to A549 cells and elastase-induced lung injury mice model. Then, various parameters such as cell-based cyto-protective activity and histopathological finding were analyzed. MCE showed a protective effect on elastase-induced cytotoxicity in A549 cells. This effect was correlated with analysis for caspase 3 levels, collagen and elastin contents, protein level of cyclin B1, Cdc2, and Erk1/2, and gene expression of TNF-${\alpha}$ and IL-$1{\beta}$ in A549 cells. MCE treatment also revealed the protective effect on elastase-induced lung injury in mice model. This effect was evidenced via histopathological finding including immunofluence stains against elastin, collagen, caspase 3, and protein level of cyclin B1, Cdc2, and Erk1/2 in lung tissue. These data suggest that MCE has a pharmaceutical properties on lung injury. This study would provide an scientific evidence for the efficacy of MCE for clinical application to patients with chronic obstructive pulmonary disease.

• Molecules and Cells
• /
• v.21 no.2
• /
• pp.251-260
• /
• 2006

During mitosis, genomic integrity is maintained by the proper coordination of anaphase entry and mitotic exit via mitotic checkpoints. In budding yeast, mitotic exit is controlled by a regulatory cascade called the mitotic exit network (MEN). The MEN is regulated by a small GTPase, Tem1p, which in turn is controlled by a two-component GAP, Bfa1p-Bub2p. Recent results suggested that phosphorylation of Bfa1p by the polorelated kinase Cdc5p is also required for triggering mitotic exit, since it decreases the GAP activity of Bfa1p-Bub2p. However, the dispensability of GEF Lte1p for mitotic exit has raised questions about regulation of the MEN by the GTPase activity of Tem1p. We isolated a Bfa1p mutant, $Bfa1p^{E438K}$, whose overexpression only partially induced anaphase arrest. The molecular and biochemical functions of $Bfa1p^{E438K}$ are similar to those of wild type Bfa1p, except for decreased GAP activity. Interestingly, in $BFA1^{E438K}$ cells, the MEN could be regulated with nearly wild type kinetics at physiological temperature, as well as in response to various checkpoint-activating signals, but the cells were more sensitive to spindle damage than wild type. These results suggest that the GAP activity of Bfa1p-Bub2p is responsible for the mitotic arrest caused by spindle damage and Bfa1p overproduction. In addition, the viability of cdc5-2 ${\Delta}bfa1 $ cells was not reduced by $BFA1^{E438K}$, suggesting that Cdc5p also regulates Bfa1p to activate mitotic exit by other mechanism(s), besides phosphorylation.