• Microbiology and Biotechnology Letters
• /
• v.21 no.3
• /
• pp.276-280
• /
• 1993

Cell lytic enzyme, 5'-phosphodiesterase, and AMP-deaminase were used to produce yeast extract as a natural seasoning from beer yeast cells. Prior to the addition of cell lytic enzyme, heat treatment was performed to increase the cell wall degradation` the optimum condition of the cell lytic enzyme was 50C at pH 7.0. The production yields by the enzymatic method and conventional autolysis method were 42% and 35%, respectively. The total quantity of 5'-nucleotides, GMP and IMP, produced by enzymatic method was increased by 45% than that by the conventional method. Futhermore, the operation time of enzymatic method was only 6.5 hrs, significantly reduced from 24 hrs of the conventional method.

• Journal of Ginseng Research
• /
• v.39 no.4
• /
• pp.354-364
• /
• 2015

Background: Intracellular $Ca^{2+}$($[Ca^{2+}]_i$) is a platelet aggregation-inducing molecule. Therefore, understanding the inhibitory mechanism of $[Ca^{2+}]_i$mobilization is very important to evaluate the antiplatelet effect of a substance. This study was carried out to understand the $Ca^{2+}$-antagonistic effect of total saponin from Korean Red Ginseng (KRG-TS). Methods: We investigated the $Ca^{2+}$-antagonistic effect of KRG-TS on cyclic nucleotides-associated phosphorylation of inositol 1,4,5-trisphosphate receptor type I ($IP_3RI$) and cyclic adenosine monophosphate (cAMP)-dependent protein kinase (PKA) in thrombin (0.05 U/mL)-stimulated human platelet aggregation. Results: The inhibition of $[Ca^{2+}]_i$ mobilization by KRG-TS was increased by a PKA inhibitor (Rp-8-BrcAMPS), which was more stronger than the inhibition by a cyclic guanosine monophosphate (cGMP)- dependent protein kinase (PKG) inhibitor (Rp-8-Br-cGMPS). In addition, Rp-8-Br-cAMPS inhibited phosphorylation of PKA catalytic subunit (PKAc) ($Thr^{197}$) by KRG-TS. The phosphorylation of $IP_3RI$ ($Ser^{1756}$) by KRG-TS was very strongly inhibited by Rp-8-Br-cAMPS compared with that by Rp-8-BrcGMPS. These results suggest that the inhibitory effect of $[Ca^{2+}]_i$ mobilization by KRG-TS is more strongly dependent on a cAMP/PKA pathway than a cGMP/PKG pathway. KRG-TS also inhibited the release of adenosine triphosphate and serotonin. In addition, only G-Rg3 of protopanaxadiol in KRG-TS inhibited thrombin-induced platelet aggregation. Conclusion: These results strongly indicate that KRG-TS is a potent beneficial compound that inhibits $[Ca^{2+}]_i$ mobilization in thrombin-platelet interactions, which may result in the prevention of platelet aggregation-mediated thrombotic disease.

• Korean Journal of Food Science and Technology
• /
• v.27 no.2
• /
• pp.241-245
• /
• 1995

Focusing on quality problems of delayed chilling porcine muscle, the effects of delayed chilling and aging on the contents of ATP-related compounds and taste of pork were investigated. Twelve Landrace pigs were employed and bisected: left sides were delay-chilled(DC) at room temperature($20^{\circ}C$) for 3 hrs, whereas right sides were conventionally chilled(CC). ATP-related compounds tested were adenosine triphosphate(ATP) and its derivatives in pork muscle, inosine monophosphate(IMP), guanosine monophosphate(GMP) and L-glutamate in cooked broth. DC sides showed more rapid pH decline and degradation of nucleotides than did CC sides. The levels of ATP and adenosine monophosphate(AMP) were not changed significantly. However, adenosine diphosphate(ADP) and IMP showed the highest levels at the 1st and 5th day, respectively. Hypoxanthine(Hx) was gradually increased(p<0.05) during aging. During aging, the IMP contents cooked broth tended to decrease, while the GMP and L-glutamate contents increase. As a result of these, the taste score got better and finally the results of sensory evaluation became increased(p<0.05). However, compared to CC sides, DC sides did not seem to lower taste of pork.

• Proceedings of the Korean Society of Applied Pharmacology
• /
• 1993.04a
• /
• pp.155-155
• /
• 1993

항우울약인 rolipran(RP)등 phosphodiesterase-억제 약(PDE-1)들이 thrombin(TB: 0.25 U/ml)에 의한 혈소판 응집에 미치는 작용을 가토-혈소판에서 일차 검토하였다. 신 PDE-1인 KR-30075(KR)의 $IC_{50}$/은 sodim nitroprusside의 것보다 낮았고 PDE-1들은 혈소판내 cAMP와 cGMP를 증가시켰으며 특히 KR은 타 PDE-1와 달리 I $P_3$를 감소시켰다. 아울러 rolipram은 cGMP와 I $P_3$를 증가시켰으나, amitriptyline(AT), sertraline(57), chlorpromazine(CP) 및 spermine은 I $P_3$를 증가시켰다. 그러나 이들과 PDE-1들은 강도의 차이는 있으나 모두 TB에 의한 혈소판응집을 모두 억제하였다. 따라서, PDE-1 중 IBMX(2$\times$$10^{-5}$M), KR(5$\times$10$_－7/M), 및 rolipram(10$_{-3}$M) 그 외에 항우울약인 AT(1.5 $\times$10$_－4/M) 와 ST(10$_－4/M) 및 항정신병약인 CP(10$_－4/M)둥이 혈소판내 I $P_3$, [C $a^{＋＋}$], Tx $B_2$, 및 PG $I_2$ 함량과 단백-인산화의 TB에 의한 변동에 미치는 영향을 검토하였다. 그 결과 TB에 의한 혈소판내 I $P_3$, [C $a^{＋＋}$], Tx $B_2$, 및 PG $I_2$ 함량의 증가가 PDE-1들과 항우울약들에 의하여 억제되었다 단, 항우울성약들과 CP는 정상 혈소판 I $P_3$를 증가시켰다. 아울러 혈소판-단백인산학에서 TB는 41-43 kD와 20 kD의 인산화를 현저히 증가시키며 19 kD의 인산화는 감소시켰고, PKC의 기질인 41-43 kD와 20 kD의 단백인산화가 PDE-1들과 항우울약들 뿐 아니라 CP에 의하여 현저히 억제되었다. 단, 20 kD 인산화에 대한 AT의 억제작용은 미약하였고, cAMP와 PDE-1들은 22kD 인산화를 증가시켰다. AT, ST, 및 CP는 A23187에 의한 41-43 kD 인산화는 현저히 억제하나 20 kD 인산화에는 영향을 미치지 않았고, PMA(3.2$\times$$10^{－7}$ M)에 의한 단백인산화에 대하여는 더 미약한 억제-효과를 나타내었다. 이상의 결과는 PDE-1과 항우울약들의 항혈소판작용은 PKC-기질인 41-43 kD와 20 kD의 인산화를 억제함에 기인되는 것으로 사료된다.다. 것으로 사료된다.다.

• KSBB Journal
• /
• v.15 no.5
• /
• pp.452-457
• /
• 2000

Due to the advantage of RP-HPLC with a variety of compositions of mobile phases, experiments on water-soluble charged species were examined. The samples were mononucleotides (5-CMP, 5-UMP, 5-GMP, 5-IMP, 5-AMP), and the buffers used were sodium phosphate monobasic and acetic acid. The concentrations of buffers ranged from 0.01 to 10 mM, while that of the methanol, an additive to the mobile phase was 5 to 20 vol.%. To predict the retention factor of a sample in terms of its methanol composition (M, vol.%) and buffer(C(sub)B, mM), the following nonlinear equation is suggested, k= $\frac{a+b C_B}{(1+c C_B) M^d}($ where a, b, c, and d were experimentally determined constants. The regression coefficients were above 0.96, and the agreement between experimental and calculated retention factors were relatively good.

• The korean journal of orthodontics
• /
• v.14 no.2
• /
• pp.187-202
• /
• 1984

There are evidences that exogenous electric currents are capable of enhancing bone formation and resolution, and that the conversion of the bioelectric response to biochemical activity provides the directional component of orthodontic tooth movement. In addition, evidence has implicated cyclic nucleotides in alveolar bone cellular activation mechanism during orthodontic tooth movement. In view of these evidences, this study was performed to investigate the effects of exogenous electric currents on cyclic nuclotide levels in feline alveolar bone and the possible clinical application of electric currents as an additional orthodontic tool. In the first study, three groups of three adult cats were subjected to application of a constant direct current of $10{\pm}2$ microamperes to gingival tissue near maxillary canine noninvasively for 1, 3, and 7 days respectively. In the second study, three groups of three adult cats each were treated by an electric-orthodontic procedure for 1, 3, and 7 days respectively. The left maxillary (control) canine received an orthodontic force of 80gm alone at time of initiation, while the right maxillary (experimental) canine received combined force-electric stimulation (80gm of force and $10{\pm}2$ microamperes of a constant D.C. currents). Alveola, bone samples were obtain from the mesial (tension and/or cathode) and the distal (compression and/or anode) sites surrounding maxillary canines as well as from contralateral control sites. The samples were extracted, boiled, homogenized, and the supernatants were assayed for cyclic nucleotides (cAMP, cGMP) by a radioimmunoassay method. And also the amount of tooth movement was measured in the second study. On the basis of this study, the following conclusions can be drawn: 1. The fluctuation pattern of cyclic nucleotide levels in alveolar bone treated by exogenous electric currents was similar to that treated by orthodontic force. 2. The cAMP levels in alveolar bone of electrically treated teeth significantly elevated above the control values. And of electrically treated teeth, the values of the anode sites were higher than those of the cathode sites. 9. The cGMP levels in alveolar bone of electrically treated teeth elevated above the control values at the initiation phase of treatment, but dropped below the control values at time of termination. And of electrically treated teeth, the values of the cathode sites were higher than those of the anode sites. 4. The rate of tooth movement in teeth . treated by force-electric combination increased with the length of treatment as compared to that treated by mechanical force alone.

• Korean Journal of Veterinary Research
• /
• v.37 no.1
• /
• pp.119-128
• /
• 1997

The relaxation of gastric fundus smooth muscles is the primary physiological event which induces the receptive relaxation of monogastric animals. L-arginine/Nitric oxide(L-arg/NO) system is known to mediate the inhibitory non-adrenergic non-cholinergic(NANC) neurotransmission in various tissues including gastrointestinal smooth muscles. The longitudinal smooth muscles of porcine gastric fundus showed fast relaxation during electrical field stimulation(EFS) and rebound contraction after EFS in NANC condition. So, the purpose of present study was elucidation of the neurotrasmitters related to the NANC relaxation and explanation of the relation between NANC relaxation and L-arg/NO system. The longitdinal smooth muscles of porcine gastric fundus were hung in the organ bath and under the presence of guanethidine($5{\times}10^{-5}M$), precontraction was induced by carbachol($1{\times}10^{-6}M$). The muscle responses to EFS and drugs were isomerically recorded. The rusults were summarized as follows. 1. The longtudinal muscles of porcine gastric fundus showed frequency-dependent relaxation and rebound contraction to electrical field stimulaton(1ms, 8V, 1~16Hz, 20sec, EFS). These responses were blocked by tetrodotoxin($1{\times}10^{-6}M$). 2. The relaxation and rebound contraction of the longitudinal muscles of porcine gastric fundus to EFS were inhibited by L-NAME($2{\times}10^{-5}M$). The inhibitory effect of L-NAME was antagonized by L-arginine($1{\times}10^{-3}M$), but not by D-arginine($1{\times}10^{-3}M$). 3. Exogenous NO($NaNO_2$, $1{\times}10^{-5}{\sim}1{\times}10^{-4}M$, pH=2.0) caused concentration-dependent relaxation as EFS did. 4. Methylene Blue($2{\times}10^{-5}M$), a soluble guanylate cyclase inhibitor, inhibited the relaxation and rebound contraction of the longitudinal muscles of porcine gastric fundus induced by EFS, but N-ethlmaleimide, a adenylate cyclase inhibitor, did not. 5. 8-Br-cGMP($1{\times}10^{-6}{\sim}3{\times}10^{-6}M$), permeable cGMP analogue, induced dose-dependent relaxation. but 8-Br-cAMP($1{\times}10^{-6}{\sim}3{\times}10^{-6}M$), permeable cAMP analogue, did not. Both did not evoked rebound contraction. 6. ${\alpha}$-chymotrypsin did not affect the relaxation of the longitudinal muscles of porcine gastric fundus. 7. Reactive blue 2($1{\times}10^{-4}M$, 40min) siginificantly inhibited the rebound contraction induced by EFS and inhibited contraction caused by exogenous ATP($1{\times}10^{-4}{\sim}1{\times}10^{-3}M$). These results suggests that NANC relaxation of the longitudinal muscles of porcine gastric fundus mainly mediated by NO and the rebound contraction is related to NO and other neurotransmitters.

• Proceedings of the Ginseng society Conference
• /
• 1998.06a
• /
• pp.216-223
• /
• 1998

Cyclic nucleotide phosphodiesterases (PDEs) represent the unique enzymatic system degrddinf cAMP and cGMP which play a major role in the regulation of cell physiology. To investigate a possible molecular mechanism of ginsenosides, their activities were evaluated on PDEs which are recently described is new therapeutic targets. PDEs are classified into 7 families according to their genes (PDEI to PDE7) and are differently distributed in tissues. The IC50 values of ginsenosides were determined on PDEI to PDE 5 chromatographically isolatetl from bovine aorta. The results show that total ginseng saponin extract preferentially inhibits PDE 1 and PDE4 at concentrations nearby 200 ug/ml. Protopanaxadiol (PPD) fraction acts preferentially on PDE4 with and IC50 value of 100 nlml and inhibits also PDEI and PDE5 at 14 to 2 fold higher concentrations, respectively. Protopanaxatriol (PPT) fraction preferentially inhibits PDE 1 with and IC50 value of 170 ug/ml. Compound Rgl, originated from PPT fraction, and RC3 (5) represent the most active compounds towards PDE 1 with IC50 values around 80 UM. However Rg3 (R), epimer of Rgl (5) has no effect on the various PDEs tested, excepted on PDE3 rich is sligthly sensitive Compound Rbl, originated from PPD, acts on both PDEI and PDE4. It if two fold less active than Rgl and Rg3 (5) on PDEI. Taken together, these results mainly suggest that PDEI and PDE4 inhibitions could be a molecular mechanism which would participate in ginsenoside mechanisms, especially the effect of PPD on blood vessel and on CNS.

• BMB Reports
• /
• v.52 no.5
• /
• pp.336-341
• /
• 2019

The cGAS-STING pathway plays an important role in pathogen-induced activation of the innate immune response. The 29-kDa amino-terminal fibronectin fragment (29-kDa FN-f) found predominantly in the synovial fluid of osteoarthritis (OA) patients increases the expression of catabolic factors via the toll-like receptor-2 (TLR-2) signaling pathway. In this study, we investigated whether 29-kDa FN-f induces inflammatory responses via the cyclic GMP-AMP synthase (cGAS)/stimulator of interferon gene (STING) pathway in human primary chondrocytes. The levels of cGAS and STING were elevated in OA cartilage compared with normal cartilage. Long-term treatment of chondrocytes with 29-kDa FN-f activated the cGAS/STING pathway together with the increased level of gamma-H2AX, a marker of DNA breaks. In addition, the expression of pro-inflammatory cytokines, including granulocyte-macrophage colony-stimulating factor (GM-CSF/CSF-2), granulocyte colony-stimulating factor (G-CSF/CSF-3), and type I interferon ($IFN-{\alpha}$), was increased more than 100-fold in 29-kDa FN-f-treated chondrocytes. However, knockdown of cGAS and STING suppressed 29-kDa FN-f-induced expression of GM-CSF, G-CSF, and $IFN-{\alpha}$ together with the decreased activation of TANK-binding kinase 1 (TBK1), interferon regulatory factor 3 (IRF3), and inhibitor protein ${\kappa}B{\alpha}$ ($I{\kappa}B{\alpha}$). Furthermore, NOD2 or TLR-2 knockdown suppressed the expression of GM-CSF, G-CSF, and $IFN-{\alpha}$ as well as decreased the activation of the cGAS/STING pathway in 29-kDa FN-f-treated chondrocytes. These data demonstrate that the cGAS/STING/TBK1/IRF3 pathway plays a critical role in 29-kDa FN-f-induced expression of pro-inflammatory cytokines.

• The Korean Journal of Physiology
• /
• v.28 no.2
• /
• pp.143-150
• /
• 1994

The whole-cell mode of the patch clamp technique was used to study $Ca^{2+}-activated\;Cl^-\;current$ $(I_{Cl_{Ca}})$ in gastric antral myocytes. Extracellular application of caffeine evoked $Ca^{2+}-activated\;current$. In order to isolate the chloride current from background current, all known systems were blocked with specific blockers. The current-voltage relationship of caffeine-induced current showed outward rectification and it reversed at around $E_{Cl^-}$. The shift of reversal potential upon the alteration of external and internal chloride concentrations was well fitted with results which were calculated by the Nernst equation. Extracellular addition of N-phenylanthranilic acid and niflumic acid which are known anion channel blockers abolished the caffeine induced current. Intracellular application of a high concentration of EGTA also abolished this current. Application of c-AMP, c-GMP, heparin, or $AIF^-_4$ made no remarkable changes to this current. Sodium replacement with the impermeable cation N-methylglucamine or with $Cd^{2+}$ rarely affected this current. From the above results it is suggested that the caffeine induced current was a $Cl^-$ current and it was activated by intracellular $Ca^{2+}$.