• The Korean Journal of Physiology
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• 제30권1호
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• pp.63-76
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• 1996

The aim of present study was to characterize phosphate uptake and to investigate the mechanism for the insulin and insulin-like growth factor(IGF) stimulation of phosphate uptake in primary cultured rabbit renal proximal tubule cells. Results were as follows : 1. The primary cultured proximal tubule cells had accumulated $6.68{\pm}0.70$ nmole phosphate/mg protein in the presence of 140 mM NaCl and $2.07{\pm}0.17$ nmole phosphate/mg protein in the presence of 140 mM KCl during a 60 minute uptake period. Raising the concentration of extracellular phosphate to 100 mM$(48.33{\pm}1.76\;pmole/mg\;protein/min)$ induced decrease in phosphate uptake compared with that in control cells maintained in 1 mM phosphate$(190.66{\pm}13.01\;pmole/mg\;protein/min)$. Optimal phosphate uptake was observed at pH 6.5 in the presence of 140 mM NaCl. Phosphate uptake at pH 7.2 and pH 7.9 decreased to $83.06{\pm}5.75%\;and\;74.61{\pm}3.29%$ of that of pH 6.5, respectively. 2. Phosphate uptake was inhibited by iodoacetic acid(IAA) or valinomycin treatment $(62.41{\pm}4.40%\;and\;12.80{\pm}1.64%\;of\;that\;of\;control,\;respectively)$. When IAA and valinomycin were added together, phosphate uptake was inhibited to $8.04{\pm}0.61%$ of that of control. Phosphate uptake by the primary proximal tubule cells was significantly reduced by ouabain treatment$(80.27{\pm}6.96%\;of\;that\;of\;control)$. Inhibition of protein and/or RNA synthesis by either cycloheximide or actinomycin D markedly attenuated phosphate uptake. 3. Extracellular CAMP and phorbol 12-myristate 13 acetate(PMA) decreased phosphate uptake in a dose-dependent manner in all experimental conditions. Treatment of cells with pertussis toxin or cholera toxin inhibited phosphate uptake. cAMP concentration between $10^{-6}\;M\;and\;10^{-4}\;M$ significantly inhibited phosphate uptake. Phosphate uptake was blocked to about 25% of that of control at 100 ng/ml PMA. 3-Isobutyl-1-methyl-xanthine(IBMX) inhibited phosphate uptake. However, in the presence of IBMX, the inhibitory effect of exogenous cAMP was not significantly potentiated. Forskolin decreased phosphate transport. Acetylsalicylic acid did not inhibit phosphate uptake. The 1,2-dioctanoyl-sn-glycorol(DAG) and 1-oleoyl-2-acetyl-sn- glycerol(OAG) showed a inhibitory effect. However, staurosporine had no effect on phosphate uptake. When PMA and staurosporine were treated together, inhibition of phosphate uptake was not observed. In conclusion, phosphate uptake is stimulated by high sodium and low phosphate and pH 6.5 in the culture medium. Membrane potential and intracellular energy levels are also an important factor fer phosphate transport. Insulin and IGF-I stimulate phosphate uptake through a mechanisms that involve do novo protein and/or RNA synthesis and decrease of intracellular cAMP level. Also protein kinase C(PKC) is may play a regulatory role in transducing the insulin and IGF-I signal for phosphate transport in primary cultured proximal tubule cells.

• Journal of Korean Vacuum Science & Technology
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• 제4권4호
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• pp.102-106
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• 2000

We report scanning force microscope (SFM) observations of enhanced calcite dissolution in aqueous solution due to mechanical stimulation induced by the SFM tip. Images and mechanical treatment were performed in saturated ($\geq$ 60 ${\mu}{\textrm}{m}$) CaCO$_3$ solution adjusted to pH~9. Small area scans of monolayer steps significantly increased the step velocity in the scanned area (in the direction corresponding to dissolution) when the applied contact force is above about 160 nN fer the tips employed. The step velocity could be increased at least an order of magnitude by scanning at even higher contact forces (e.g.,270nN). This enhancement is a function of step orientation relative to the calcite lattice. Indentations near preexisting steps also locally enhance the step velocity. We present evidence that the higher dissolution rates are caused by stress-induced increases in the rate of double-kink nucleation.

• 대한치과의사협회지
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• 제11권1호
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• pp.45-52
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• 1973

Ketamine HC1(Phencyclidine derivative) is a white, crystalline substance with melting point 259℃. It is soluble in water to a 20% clear, colorless solution. a 10% aqueous solution has a pH 3.5. The chemical structure is 2-(O-chlorophenyl)-2-(methylamino) cyclohexanone hydrochloride. It is a rapid acting, nonhypnotic, nonbarbiturate drug with a wide safety margin. The author used the anesthetic ketamine hydrochloride for oral surgery procedure in 16 patients(2 to 33 years). Anesthesia was achieved with a single intramuscular injection by introduction of manufacture. Result obtained as follows : 1. It was easily administered. 2. Onset time was rapid and duration was short. 3. The mild cardiac stimulation resulted in moderate increase in the blood pressure and pulse. 4. There was no significant respiratory depression. 5. The airway can be maintained without artificial support or endotracheal intubation. 6. All protective reflex maintained, which was important in oral surgery. 7. The best results were achieved in pediatric patients than adults.

• 대한본초학회지
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• 제32권2호
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• pp.107-114
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• 2017

Objectives : Tribulus terrestris $Linn{\acute{e}}$ (Tribuli Fructus; TF) has been used to treat hypochondrium, agalactia, nebula, itching and vitiligo in traditional Korean medicine. In this study, we investigated the effects of TF 30% ethanol extract on inflammatory responses in IgE-stimulated RBL-2H3 mast cells. Methods : TF extract was prepared by 30% ethanol. RBL-2H3 cells, a rat mast cell line, were treated with TF extract at different concentrations for 1 hr and then stimulated with DNP-IgE/HSA for indicated times. Cell viability was measured by WST-1 assay. The expression of inflammatory cytokines (IL-4, IL-13 and $IFN-{\gamma}$) mRNA was determined by reverse transcriptase-PCR, and the phosphorylation of ERK1/2, p38 and JNK MAP kinases (MAPKs) was determined by Western blot. The nuclear expression of $NF-{\kappa}B$ p65 in the cells was detected by Western blot and immunocytochemistry, respectively. Results : The treatment of TF extract at 0.1 and $0.2mg/m{\ell}$ significantly decreased the expression of IL-4 and IL-13 mRNA in IgE-stimulated RBL-2H3 mast cells, while significantly increased the expression of $IFN-{\gamma}$ mRNA. TF extract treatment was also inhibited the phosphorylation of ERK1/2, p38 and JNK MAPKs in IgE-stimulated RBL-2H3 mast cells in a dose-dependent manner. In addition, TF extract significantly blocked the translocation of $NF-{\kappa}B$ p65 into the nuclear of cells after IgE stimulation. Conclusions : These results indicate that TF extract inhibits inflammatory response in IgE-stimulated mast cells through blocking MAPKs/$NF-{\kappa}B$ pathway. This suggests that TF extract has an anti-inflammatory activity in mast cell activation.

• Journal of Ginseng Research
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• 제30권2호
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• pp.88-94
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• 2006

인삼과 유산균은 각각 면역 활성, 생리기능조절 등 생체 효용성을 인정받고 있는 가능성 식품소재이다. 따라서 유산균을 이용한 인삼 발효물은 두 소재의 장점을 모두 이용할 수 있는 가능성이 있으므로 본 연구는 여러 가지 유산균을 이용하여 인삼 발효물을 제조하고 생균수, pH 및 산도 등을 조사하여 인삼 발효물 제조에 적합한 최적의 유산균주를 선정하고 인삼의 발효공정을 확립하였다. 27종의 유산균 중 인삼농도가 증가할수록 생육이 촉진되는 유산균은 Lac.plantarum MG208, Lac.casei Mg311, Lac.rhamnosus, MG315, Lac.acidophilus MG501, Lac.acidophilus MG501C, Lac.reuteri MG505, Lac.fermentum MG590, Bif.longum MG723 등 8종이었다. 특히, Lac.plantarum MG208이 5% 홍삼배지에 배양했을 때 가장 생육이 촉진되는 균주였다. 발효 전 배지 pH를 6.8로 동일하게 보정하였을 때 최종 인삼발효물의 pH는 유의하게 낮아졌고($pH3.45{\sim}4.83$), 특히, 홍삼농도가 높을수록 pH는 더 낮았다($pH3.42{\sim}4.30$). 이와 유사하게 백삼 및 홍삼 발효제조물의 적정산도는 약 10배정도 증가했고, 적정산도 증가율이 가장 높은 균종은 Lac.plantarum MG208, Lac.acidophilus MG501, 및 Bif.longum MG723이었으며, 발효 후 적정산도는 백삼보다 홍삼을 첨가한 경우 좀 더 높았다. 이와같이 여러 가지 유산균주를 이용하여 인삼 발효물을 제조한 결과 생균수와 유산균 발효제품에 적용할 수 있는 pH 및 적정산도 유도면에서 발효인삼 제조에 가장 적합한 유산균주는 Lac.plantarum MG208이며 인삼첨가농도는 1-5%, 배양시간은 48시간이 적합한 것으로 생각된다. 인삼의 유산균 발효물의 생리활성 효능은 앞으로 추가연구를 통하여 검증되어야 하며 그 유용성에 대한 고찰 및 검토가 필요하다.

• Journal of Korean Neurosurgical Society
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• 제55권5호
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• pp.237-243
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• 2014

Objective : Symptomatic disc degeneration develops from inflammatory reactions in the annulus fibrosus (AF). Although inflammatory mediators during annular inflammation have been studied, the roles of matrix metalloproteinases (MMPs) and their inhibitors have not been fully elucidated. In this study, we evaluated the production of MMPs and tissue inhibitors of metalloproteinase (TIMPs) during annular inflammation using an in vitro co-culture system. We also examined the effect of notochordal cells on annular inflammation. Methods : Human AF (hAF) pellet was co-cultured for 48 hours with phorbol myristate acetate-stimulated macrophage-like THP-1 cells. hAF pellet and conditioned media (CM) from co-cultured cells were assayed for MMPs, TIMPs, and insulin-like growth factor (IGF)-1 levels using real-time reverse-transcriptase polymerase chain reaction and enzyem-linked immunosorbent assay. To evaluate whether notochordal cells affected MMPs or TIMPs production on annular inflammation, hAF co-cultured with notochordal cells from adult New Zealand White rabbits, were assayed. Results : MMP-1, -3, -9; and TIMP-1 levels were significantly increased in CM of hAF co-cultured with macrophage-like cells compared with hAF alone, whereas TIMP-2 and IGF-1 levels were significantly decreased (p<0.05). After macrophage exposure, hAF produced significantly more MMP-1 and -3 and less TIMP-1 and -2. Interleukin-$1{\beta}$ stimulation enhanced MMP-1 and -3 levels, and significantly diminished TIMP-2 levels. Co-culturing with rabbit notochordal cells did not significantly influence MMPs and TIMPs production or COL1A2 gene expression. Conclusion : Our results indicate that macrophage-like cells evoke annular degeneration through the regulation of major degradative enzymes and their inhibitors, produced by hAF, suggesting that the selective regulation of these enzymes provides future targets for symptomatic disc degeneration therapy.

• Tuberculosis and Respiratory Diseases
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• 제49권3호
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• pp.332-342
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• 2000

연구배경 : 염증매개 사이토카인은 염증성 폐질환의 중요한 매개물질이다. 폐 상피세포는 염증세포에서 분비되는 사이토카인에 의해 interleukin, chemokines, colony stimulating factors와 growth factor등을 생산 및 분비함으로써 국소 염증 부위에서의 사이토카인 network에 중요한 역할을 한다. 따라서 폐 상피세포에서 염증매개 사이토카인의 발현 기전에 대한 이해는 염증성 폐질환의 기전규명과 이에 기초한 새로운 치료법의 개발에 생각된다. 대부분의 사이토카인은 NF-${\kappa}B$전사인자에 의해 발현되는데 폐 상피세포에서 염증매개 사이토키인의 발현과 NF-${\kappa}B/I{\kappa}B$ 경로와의 관련성에 관한 연구는 부족한 실정이다. 방법 : BEAS-2B, A549, NCI-H157, NCI-H719 세포에서 IL-1$\beta$와 TNF-$\alpha$ 자극에 의한 IL-8과 TNF-$\alpha$ mRNA의 발현 양상을 평가하였고 이들의 발현과 관찰하였고 NF-${\kappa}B/I{\kappa}B$ 경로와의 관련성을 평가하기 위하여 IL-l$\beta$와 TNF-$\alpha$ 자극에 의한 NF-${\kappa}B$의 활성화 및 $I{\kappa}B{\alpha}$와 $I{\kappa}B{\beta}$의 분해 양상을 관찰하였다. 폐 상피세포의 종류에 따른 NF-${\kappa}B/I{\kappa}B$ 경로 조절의 기전을 규명하고자 IL-1$\beta$와 TNF-$\alpha$ 자극에 의한 $I{\kappa}B{\alpha}$의 인산화와 기저상태에서 IKK$\alpha$의 발현을 평가하였다. 결과 : BEAS-2B, A549, NCI-H157 세포에서는 IL-1$\beta$와 TNF-$\alpha$ 자극으로 $I{\kappa}B{\alpha}$와 $I{\kappa}B{\beta}$가 분해되었고 NF-${\kappa}B$의 활성화가 관찰되었으며 IL-8과 TNF-$\alpha$mRNA의 발현이 유도되었다. NCI-H719 세포에서는 IL-1$\beta$와 TNF-$\alpha$ 자극으로 $I{\kappa}B$ 분해에 의한 NF-${\kappa}B$의 활성화 및 염증매개 사이토카인의 발현이 관찰되지 않았다. BEAS-2B, A549, NCI-H157 세포에서는 IL-1$\beta$와 TNF-$\alpha$ 자극으로 ${\kappa}B$의 인산화가 관찰되었지만 NCI-H719 세포에서는 관찰되지 않았다. 기저상태의 IKK$\alpha$ 발현은 세포간에 차이가 관찰되지 않았다. 결론 : 폐 상피세포에서 NF-${\kappa}B/I{\kappa}B$ 경로는 염증매개 사이토카인 발현에 매우 중요한 역할을 하고, 일부 세포에서는 NF-${\kappa}B/I{\kappa}B$ 경로 조절의 차이를 보이는데 이는 IKK보다 상위 단계의 세포내 신호전달체계의 이상에 기인한 것으로 생각된다.

• 대한의생명과학회지
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• 제19권4호
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• pp.344-347
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• 2013

The S100A8 and S100A9 proteins play important roles in inflammatory diseases. The house dust mite acts as a major allergen that induces allergic diseases. We investigated the effect of the house dust mite on S100A8 and S100A9 protein expression in monocytes. We also examined the effect of CCL2, a powerful monocyte chemoattractant, on the expression of both proteins. Extract of Dermatophagoides pteronissinus (DP), recombinant Der p 1 and Der p 2, or CCL2 had no effect on S100A8 and S100A9 expression in human monocytic THP-1 cells. Monocytes were isolated from healthy donors and treated with DP, Der p 1, and Der p 2. S100A8 expression in monocytes increased after a 24 h stimulation with DP, Der p 1, and Der p 2, and CCL2 also increased S100A8 production. However, S100A9 expression in monocytes was not altered by DP, Der p 1, Der p 2, or CCL2. These results indicate that house dust mite and CCL2 may trigger an inflammatory response by altering S100A8 expression.

• Journal of Microbiology
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• 제42권2호
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• pp.117-125
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• 2004

Propionibacterium acnes (P. acnes) plays an important role in the disease pathogenesis of acne vulgaris, a disorder of pilosebaceous follicles, seen primarily in the adolescent age group. In the present study, the presence of antibodies against P. acnes (MTCC1951) were detected in acne patient (n=50) and disease free controls (n=25) using dot-ELISA and Western blot assay. The ability of P. acnes to induce pro-inflammatory cytokines by human peripheral blood mononuclear cells (PBMCs), obtained from acne patients and healthy subjects, were also analysed. The patients (n=26) who were culture positive for skin swab culture, were found to have a more advanced disease and higher antibody titres (1:4000 to >1:16000) compared to the P. acnes negative patients (n=24) and normal controls (n=25). An analysis of patients' sera by western blot assay recognized a number of antigenic components of P. acnes, rang-ing from 29 to 205 kDa. The major reactive component was an approximately 96 kDa polypeptide, which was recognised in 92％ (24 of 26) of the patients sera. Further, the P. acnes culture supernatant, crude cell lysate and heat killed P. acnes whole cells, obtained from 72-h incubation culture, were observed to be able to induce significant amounts of IL-8 and tumor necrosis factor alpha (TNF-${\alpha}$) by the PBMCs in both the healthy subjects and patients, as analysed by cytokine-ELISA. The levels of cytokines were significantly higher in the patients than the healthy subjects. A major 96 kDa polypep-tide reactant was eluted from the gel and was found to cause dose dependent stimulation of the pro-ductions of IL-8 and TNF-${\alpha}$. Thus, the above results suggest that both humoral and pro-inflammatory responses play major roles in the pathogenesis of acne.

• Biomolecules & Therapeutics
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• 제13권2호
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• pp.101-106
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• 2005

The generation of secretory IgA antibodies (Abs) for specific immune protection of mucosal surfaces depends on stimulation of the mucosal immune system, but this is not effectively achieved by parenteral or even oral administration of most soluble antigens. Thus, to produce a possible vaccine antigen against urinary tract infections, the uropathogenic E. coli (UPEC) adhesin was genetically coupled to the heat-labile Escherichia coli enterotoxin A2B (ltxa2b) gene and cloned into a pMAL-p2E expression vector. The chimeric construction of pMALfimH/ltxa2b was then transformed into E. coli K-12 TB1 and its nucleotide sequence was verified. The chimeric protein was then purified by applying the affinity chromatography. The purified chimeric protein was confirmed by SDS-PAGE and westem blotting using antibodies to the maltose binding protein (MBP) or the heat labile E. coli subunit B (LTXB), plus the N-terminal amino acid sequence was analyzedd. The orderly-assembled chimeric protein was confirmed by a modified $G_{M1}$-ganglioside ELISA using antibodies to adhesin. The results indicate that the purified chimeric protein was an Adhesin/LTXA2B protein containing UPEC adhesin and the $G_{M1}$-ganglioside binding activity of LTXB. thisstudy also demonstrate that peroral administration of this chimeric immunogen in mice elicited high level of secretory IgA (sIgA) and serum IgG Abs to the UPEC adhesin. The results suggest that the genetically linked LTXA2B acts as a useful mucosal adjuvant, and that adhesin/LTXA2A chimeric protein might be a potential antigen for oral immunization against UPEC.