• The Korean Journal of Zoology
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• v.38 no.1
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• pp.78-86
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• 1995

Our previous report has suggested that the decrease of fibronectin level during mvogenesis is due to the decreased Bvailabilitv of receptor (matrix assembly receptor) for 29-kDa fragment of fibronectin. In the present study, we demonstrate that G protein and adenvlate cvclase system are involved in the regulation of fibronectin matrix assembly and that when fibronectin level in extracellular matrix decreases, the postmitotic fusion-capable cells emerge more frequently from the proliferative population. This proposal is based on the following observations. (1) Cholers toxin, which increases intracellular CAMP, caused a decrease in the ability of mvoblasts to incorporate fibronectin into extracellular matrix. (2) Cholera toxin decreased the proliferation of mvoblasts and Induced the precocious fusion. (3) decAMP, which was found to induce the precocious fusion and decrease the proliferation of myoblasts, decreased the fibronectin level in extracellular matrix and matrix assembly receptor for fibronectin- (4) RGOS, whlh inhibits the incorporation of fibronectin into extracellular matrix, induced the precocious fusion and reduced the proliferaton of mvoblasts. These results suggest that CAMP regulates the fibronectin levels in extracellular matrix and that the alteration of fibronectin level is involved in regulation of the proliferation and differentiation of chick embryonic mvoblasts.

• International Journal of Oral Biology
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• v.30 no.1
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• pp.9-15
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• 2005

Recently, we reported that high extracellular calcium increased receptor activator of nuclear factor-${\kappa}B$ ligand (RANKL) expression via p44/42 mitogen-activated protein kinase (p44/42 MAPK) activation in mouse osteoblasts. However, the mechanism for p44/42 MAPK activation by high extracellular calcium is unclear. In this study, we examined the role of intracellular calcium increase in high extracellular calcium-induced RANKL induction and p44/42 MAPK activation. Primary cultured mouse calvarial osteoblasts were used. RANKL expression was highly induced by 10 mM calcium treatment. Ionomycin, a calcium ionophore, also increased RANKL expression and activated p44/42 MAPK. U0126, an inhibitor of MEK1/2, an upstream activator of p44/42 MAPK, blocked the RANKL induction by both high extracellular calcium and ionomycin. High extracellular calcium increased the phosphorylation of proline-rich tyrosine kinase 2 (Pyk2), one of the known upstream regulators of p44/42 MAPK activation. Bisindolylmaleimide, an inhibitor of protein kinase C, did not block RANKL induction and p44/42 MAPK activation induced by high extracellular calcium. 2-Aminoethoxydiphenyl borate, an inhibitor of inositol 1,4,5-trisphosphate (IP3) receptor, blocked the RANKL induction by high extracellular calcium. It also partially suppressed the activation of Pyk2 and p44/42 MAPK. Cyclosporin A, an inhibitor of calcineurin, also inhibited high calcium-induced RANKL expression in dose dependent manner. However, cyclosporin A did not affect the activation of Pyk2 and p44/42 MAPK by high extracellular calcium treatment. These results suggest that 1) the increase in intracellular calcium via IP3-mediated calcium release is necessary for RANKL induction by high extracellular calcium treatment, 2) Pyk2 activation, but not protein kinase C, following the increase in intracellular calcium might be involved in p44/42 MAPK activation, and 3) calcineurin-NFAT activation by the increase in intracellular calcium is involved in RANKL induction by high extracellular calcium treatment.

• Korean Journal of Microbiology
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• v.27 no.4
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• pp.373-377
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• 1989

The effects of environmental factors on the extracellular release of organic carbon by Scenedesmus qudricauda were studied. The PER (percentage extracellular release) was greater at high temperature and at high concentration of nitrogen and phosphate. The PER variation according to the change of M/P ratio showed high values at each extreme N/P ratio. This result suggested that the limitation of nitrogen or phosphorous resulted in the accumulation of carbohydrates as photosynthetic products, and the products in high concentration were excreted through algal cell membrane.

• Microbiology and Biotechnology Letters
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• v.27 no.3
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• pp.203-207
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• 1999

V. alginolyticus 138-2, a marine halophilic bacterium, produced an extracellular amylase with a molecular weight of ca. 56,000. The analysis of the digestion products of soluble starch by thin layer chromatography(TLC) revealed that the extracellular amylase of V. alginolyticus 138-2 is a saccharifying-type alpha-amylase. The alpha-amylase activity of the culture supernatant of soluble starch was optimal at pH 6.0 and 45$^{\circ}C$. Ca2+ slightly increased the alpha-amylase activity, whereas Hg2+, An2+, Cu2+, Ni2+, Fe2+, and Mn2+inhibited the enzymatic activity. Alkylating thiol group agent, iodoacetic acid did not affect the alpha-amylase activity, but reduced thiol reagents such as dithiothreitol, cysteine, and beta-mercaptoethanol stimulated theenzymatic activity. On the other hand, even if V. alginolyticus 138-2 is a marine halophilic bacterium, its alpha-amylase activity was significantly inhibited by NaCl.

• Biotechnology and Bioprocess Engineering:BBE
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• v.10 no.6
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• pp.546-551
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• 2005

[ ${\beta}-1,3-Glucan$ ] (curdlan) is a water-insoluble polysaccharide composed exclusively of ${\beta}-1,3\;linked$ glucose residues. Extracellular curdlan was mostly synthesized by Agrobacterium species and Alcaligenes faecalis under nitrogen-limiting conditions. In this study, we screened the microorganisms capable of producing extracellular curdlan from soil samples. For the first time, we reported Gram-positive bacterium Bacillus sp. SNC 107 capable of producing extracellular curdlan in appreciable amounts. The effect of different carbon sources on curdlan production was studied and found that the yield of curdlan was more when glucose was used as carbon source. It was also found that maximum production was achieved when the initial concentration of ammonium and phosphate in the medium was 0.5 and 1.9 g/L respectively. In this study the curdlan production was increased from 3 to 7g/L in shake flask cultures.

• Biotechnology and Bioprocess Engineering:BBE
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• v.3 no.2
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• pp.109-111
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• 1998

The effect of dissolved oxygen concentration on the metabolism of glucose in Pseudomonas putida BM014 was investigated. Glucose was completely converted to 2-ketogluconate via extracellular oxidative pathway and then taken up for cell growth under the condition of sufficient dissolved oxygen concentration. On the other hand, oxygen limitation below dissolved oxygen tension (DOT) value of 20% of air saturation caused the shift of glucose metabolism from the extracellular oxidative pathway to the intracellular phosphorylative pathway. Specific activities of hexokinase and gluconate kinase in intracellular phosphorylation pathway decreased as the DOT increased, while 2-ketogluconokinase activity in extracellular oxidative pathway increased under the same condition. This result can be usefully applied to microbial transformation of glucose to 2-ketogluconate, the synthetic precursor for iso-vitamine C, with almost 100% yield via extracellular oxidation by simple DOT control.

• Molecules and Cells
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• v.19 no.2
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• pp.232-238
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• 2005

Corneal endothelial cells play an important role in maintaining the transparency and ionic balance of the cornea. Inflammation causes many changes in the intracellular and extracellular environment of the cornea, including acidosis. We examined the relationship between changes in extracellular pH and expression of cyclooxygenase-2 in cultured bovine corneal endothelial cells. When extracellular pH ($[pH]_o$) was reduced to pH 6.4, COX-2 mRNA increased, with a peak at 2 h. This was blocked by pretreatment with actinomycin D and incubation with spermine NONOate (SPER/NO, a nitric oxide donor). Exposure to the $H^+$ ionophore, carbonyl cyanide m-chlorophenylhydrazone (CCCP), also raised COX-2 mRNA levels. CCCP-induced COX-2 mRNA expression was also reduced by SPER/NO. These results were confirmed immuno-cytochemically. These data demonstrate that COX-2 expression is stimulated by the lowering of extracellular pH that could result from bacterial infection, and that this is countered by over-production of nitric oxide, which could also result from bacterial infection.

• Clinical and Experimental Pediatrics
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• v.48 no.6
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• pp.580-587
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• 2005

New evidence is emerging that airway smooth muscle(ASM) may act as an immunomodulatory cell by providing pro-inflammatory cytokines and chemokines, polypeptide growth factors, extracellular matrix proteins, cell adhesion receptors and co-stimulatory molecules. ASM can promote the formation of the interstitial extracellular matrix, and potentially contribute to the alterations within the extracellular matrix in asthma. In addition, extracellular matrix components can alter the proliferative, survival, and cytoskeletal synthetic function of ASM cells through integrin-directed signaling. Increased ASM mass is one of the most important features of the airway wall remodeling process in asthma. Three different mechanisms may contribute to the increased ASM mass : cell proliferation, increased migration and decreased rate of apoptosis. The major signaling pathways of cell proliferation activated by ASM mitogens are those dependent on extracellular signal-regulated kinase and phosphoinositide 3'-kinase. The key signaling mechanisms of cell migration have been identified as the p38 mitogen-activated protein kinase and the p21-activated kinase 1 pathways. ASM cells contain ${\beta}2$-adrenergic receptors and glucocorticoid receptors. They may represent a key target for ${\beta}2$-adrenergic receptor agonist/corticosteroid interactions which have antiproliferative activity against a broad spectrum of mitogens.

• Applied Biological Chemistry
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• v.31 no.4
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• pp.382-386
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• 1988

Extracellular protein produced by Bacillus sp. $T_2-3$ was characterized for its patterns of sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and sephadex G-100 filtration, spectrum of maximum absorption, composition of amino acid and solubility to solvents. The extracellular protein was composed of two kinds of protein which was little difference in molecular weight about 49,000. Maximum absorbance of the extracellular protein was showed at 230nm and main amino acids of the extracellular protein were aspartic acid, glutamic acid and alanine. Solubility of the extracellular protein was 55.8% in $H_2O$ and 28.4% in 0.4% NaOH.

• The Journal of Korean Medicine
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• v.25 no.4
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• pp.1-7
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• 2004

Objective : This study was designed to examine the effects of orally administered Prunus persica water extract (PPE), which is used as herbal medicine, for treatment of Yu Xue (stasis of blood) and tacrine on the basal concentration of extracellular acetylcholine in the hippocampus of rats. Methods: To investigate the effects of PPE and tacrine on concentration of extracellular acetylcholine in the hippocampus of rats, the microdialysis technique, under the same experimental conditions, was used. And we used male Wistar rats which were 7 weeks of age and 210-290 g. PPE was extracted with boiling water, and the rats were anesthetized with pentobarbital Na. Their skulls were exposed and a hole was drilled for implantation of a microdialysis probe. In order to increase the recovery of acetylcholine, a probe with a long membrane was used. One day after surgery, the microdialysis probe was perfused with Ringer's solution at a flow rate of 1.5 l/min. The acetylcholine concentration in dialysis samples was measured by high-performance liquid chromatography (HPLC) with electrochemical detection. AChE activity was measured using the radiometric method, as described by Sherman. Results : The comparative effects of PPE and tacrine on hippocampal extracellular acetylcholine concentration was that these cholinesterase inhibitors produced dose-dependent increases in the extracellular acetylcholine concentration. And the effect of PPE and tacrine on rat brain AChE activity was that PPE produced maximal inhibition at 1 h after administration, when AChE activity was 44% of the intact level. AChE activity gradually recovered thereafter, and reached 78% of the intact level at 12 h after administration. Conclusion : In this study, PPE has a potent activity and a long-lasting effect on the central cholinergic system, in terms of the basal concentration of extracellular acetylcholine in the hippocampus and the AChE activity in the brain of rats. And oral administration of PPE increased dose-dependently the basal concentration of extracellular acetylcholine in the hippocampus of rats. PPE may be one of the more useful cholinesterase inhibitors for the treatment of Alzheimer's disease.