• Title/Summary/Keyword: intracellular $Mg^{2+}$

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Immunopotentiating Effect of Polysaccharides Extracted from Agrocybe cylindracea (Agrocybe cylindracea로부터 추출한 다당류의 면역증강활성)

  • 김선희;이항우;배만종;이재성
    • Korean Journal of Microbiology
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    • v.36 no.1
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    • pp.64-68
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    • 2000
  • The immunopotentiating effects of the polysaccharides, both intracellular and extracellular, was examined by an animal feeding test. The results are summarized as follows. The oral administration of intracellular and extracellular polysaccharides of Agrocybe cylindracea for 10 days resulted in the enhanced phagocytic activity of peritoneal exudate cells(PEC), spleen cells(SC), and monolymphocytes(ML). In the experiment of PFC(plaque forming cell) and RFC(rosette forming cell), the results showed that all the polysaccharide fractions enhanced the immune related cells. The EAC II group(the extracellular polysaccharide of Agrocybe cylindracea 10 mg/0.2 ml distilled water/day/ mouse) increased the PFC and RFC by 46~50% and 43%, respectively, compared to the control group. On the other hand, the IAC I group(the intracellular polysaccharide of Agrocybe cylindracea 1 mg/0.2 ml distilled water/day/mouse) increased the PFC and RFC by 49~70% and 91%, respectively. In terms of the mitogenic activity, the extracellular polysaccharides of A. cylindracea showed a higher activity than the intracellular polysaccharides.

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Regulation of circulating Mg2+ in the rat by metabolic inhibition (흰쥐에서 대사작용 억제에 의한 혈중 Mg2+ 조절)

  • Kim, Jong-shick;Kim, Shang-jin;Kim, Jin-shang
    • Korean Journal of Veterinary Research
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    • v.39 no.1
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    • pp.70-76
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    • 1999
  • Magnesium ($Mg^{2+}$) plays an important role in the regulation of a range of intracellular processes. Regulation of extracellular $Mg^{2+}$ contents was studied in the anesthetized Sprague-Dawley (SD) rats. Animals were injected intraperitoneally with sodium nitrite ($NaNO_2$), and circulating $Mg^{2+}$($[Mg^{2+}]c$) was measured after the injection and then 10 and 20 minutes later. A dose-dependent increase in $[Mg^{2+}]c$ was observed in animals injected with $NaNO_2$ at a dose of 10mg/kg or higher. Pretreatment with methylene blue prevented the $NaNO_2$-induced increase in $[Mg^{2+}]c$. $[Mg^{2+}]c$ displayed an inverse linear correlation with hemoglobin and exponential correlation during $NaNO_2$ injection. Injection of KCN or rotenone also induced an increase in $[Mg^{2+}]c$. An increase in $[Mg^{2+}]c$ was observed when respiration rate was reduced from 100/min (140ml/min) to 10/min (14ml/min) during 30 min. These results indicate that changes in $[Mg^{2+}]c$ inversely reflect alteration of ATP in a model of metabolic inhibition.

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Differential Effects of Fumonisin $B_1$ on Cell Death in Cultured Cells: the Significance of the Elevated Sphinganine

  • Yu, Chang-Hun;Lee, Yong-Moon;Yun, Yeo-Pyo;Yoo, Hwan-Soo
    • Archives of Pharmacal Research
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    • v.24 no.2
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    • pp.136-143
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    • 2001
  • Fumonisins are specific inhibitors of ceramide synthase in sphingolipid metabolism. An alteration in sphingolipid metabolism as a result of fumonisin exposure is related to cell death (Yoo et al., 1992). The objective of this study was to investigate whether elevated free sphinganine levels are related to the sensitivity of cultured cells to fumonisin exposure. Fumonisin $B_1$ elevated the intracellular free sphinganine concentraions in both LLC-$PK_1$ and Chinese hamster ovary (CHO) cells. However, CHO cells are resistant to fumonisin cytotoxicity at 50${u}m$, while LLC-$PK_1$ cells are sensitive at concentrations greater than 357M. The intracellular concentration of free sphinganine in LLC-$PK_1$ cells treated at 50${u}m$ fumonisin $B_1$ for 72 h was approximately 1450 pmol/mg protein relative to the 37 pmol observed in the control culture. Under the same conditions, the population of apoptotic cells in the 50${u}m$ fumonisin $B_1$-treated culture was approximately 37% of the total compared to 12% in the control. The caspase III-like activity after 72 h in the 50${\mu}$M fumonisin $B_1$-exposed culture Increased to approximately 50 $pmol/mg$ protein/hr compared to 6 $pmol/mg$ protein/hr in the control. L-cycloserine, a serine palmitoyltransferase inhibitory reduced the fumonisin $B_1$-stimulated caspase III-like activity down to the control level. Under the same culture conditions, the intracellular concentration of free sphinganine after-cycloserine plus fumonisin $B_1$ treatment was 140 pmol/mg protein compared to 1450 $pmol/mg$ protein in fumonisin $B_1$ alone. The intracellular concentration of free sphinganine in CHO cells treated with 50${u}m$ fumonisin $B_1$ for 72 h was al)proximately 460 pmol/mg protein, indicating that the mass amount of elevated free sphinganine in the CHO cells was about 32% of that in LLC-$PK_1$ cells. Adding exogenous sphinganine to the CHO cells along with 50${u}m$ fumonisin $B_1$ treatment for 72 h caused both necrosis and apoptosis. In conclusion, the elevated endogenous sphinganine acts as a contributing factor to the fumonisin-induced cell death.

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Effect of Magnesium Sulfate on Product Inhibition of Sisomicin Production

  • Shin, Chul-Soo;Han, Sang-Hun
    • Journal of Microbiology and Biotechnology
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    • v.5 no.2
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    • pp.96-99
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    • 1995
  • Addition of l00mM $MgSO_4$ to a cell culture after 54 hours resulted in a 2.4-fold increase in the sisomicin titre compared to a control to which no $MgSO_4$ was added, and a considerable amount of intracellular sisomicin was liberated outside the cells. The occurrence of product inhibition in fermentation was confirmed by a reduction in net sisomicin production with increasing amounts of added sisomicin without addition of $MgSO_4$. All added sisomicin was bound to sisomicin-free cells in the absence of $MgSO_4$, whereas approximately 40% of added sisomicin was bound with the addition of l00mM $MgSO_4$. Under conditions of no enzmye synthesis, maintained by adding chloramphenicol to exclude product repression, sisomicin was produced in the presence of 100 mM $MgSO_4$ but little sisomicin was produced in the absence of $MgSO_4$.

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Overproduction and Secretion of $\beta$-Glucosidase in Bacillus subtilis

  • Kim, Jeong-Hyun;Lee, Baek-Rak;Moo, young-Pack
    • Journal of Microbiology and Biotechnology
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    • v.8 no.2
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    • pp.141-145
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    • 1998
  • Overproduction of intracellular ${\beta}$-glucosidase was attempted by modifying the promoter region of a ${\beta}$-glucosidase gene cloned from Cellulomonas fimi and expressing it in Bacillus subtilis DB 104. A strong engineered promoter, BJ27UΔ88, was fused to the ${\beta}$-glucosidase gene after removing its native promoter. An effective Shine-Dalgamo sequence (genel0 of phage T7) was inserted between the promoter and the ${\beta}$-glucosidase structural gene. The modified gene was overexpressed in B. subtilis and produced 1121.5 units of ${\beta}$-glucosidase per mg protein which is about $12\%$ of total intracellular protein. Secretion of overproduced intracellular ${\beta}$-glucosidase was attempted by using the signal sequence of the Bacillus endoglucanase gene as well as an in-frame hybrid protein of endoglucanase. The hybrid protein was normally secreted into the culture medium and still retained ${\beta}$-glucosidase activity.

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Effect of Chlorination on Disinfection Byproducts Production and Release of Microcystins from Bloom-forming Algae (녹조현상 원인조류들의 염소처리에 의한 소독부산물 생성 및 microcystins 유출)

  • Park, Hae-Kyung;Seo, Yong-Chan;Cho, Il-Hyung;Park, Byung-Hwang
    • Journal of Korean Society on Water Environment
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    • v.22 no.3
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    • pp.513-520
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    • 2006
  • The effect of chlorination on disinfection byproducts (DBPs) production from bloom-forming freshwater algae including 7 strains of cyanobacteria and 6 strains of diatoms was investigated. The release and degradation of hepatotoxin (microcystins) by the chlorination on Microcystis under differential condition of the chlorination time and dose were also investigated. The disinfection byproducts formation potentials (DBPFP) of cyanobacterial species and diatoms were ranged from 0.017 to $0.070{\mu}mol\;DBPs/mg$ C and from 0.129 to $0.708{\mu}mol\;DBPs/mg$ C respectively. Among three major groups of DBPs, haloacetonitrils (HANs) was major product in most test strains except Aphanizomenon sp. and Oscillatoria sp. Haloacetic acids (HAAs) was less than 5 % of total DBPs. Chloroform and dichloroacetonitril (DCAN) were dominant compounds in trihalomethanes (THMs) and HANs respectively. After 4 hours chlorination of toxic Microcystis aeruginosa under the dose range of 0.5 to $10mg\;Cl_2/L$, the concentration of intracellular microcystins decreased, but dissolved dissolved microcystins concentration increased with the treatment of more than $3mg\;Cl_2/L$. However the total amount of microcystins was almost constant even at $10mg\;Cl_2/L$ of chlorination. To conclude, our results indicate that the chlorination causes algal cell lysis and release of intracellular microcystins in the intact form to surrounding waters.

Effects nit Mineral Salts on the Improvements of Sisomicin field (무기질 염이 Sisomicin 발효 수율의 증가에 미치는 영향)

  • Shin, Chul-S;Sang H. Han;Lee, Sang H.
    • Microbiology and Biotechnology Letters
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    • v.17 no.3
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    • pp.247-251
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    • 1989
  • Effects of mineral salts on sisomicin fermentation were investigated. The optimal concentration of CoCl$_2$for accomplishing a high antibiotic yield was found to be 16.8 $\mu$M at which it could function as a cofactor. At this level the other mineral salts tested had no effect. On the other hand, at much higher concentration levels (above 1 mM), four mineral salts such as ZnSO$_4$, KH$_2$PO$_4$, FeSO$_4$and MgSO$_4$were used in order to liberate the intracellular sisomicin out-side the cells, because the sisomicin accumulated mostly in cells and it was supposed to limit the improvement of antibiotic yield. ZnSO$_4$and KH$_2$PO$_4$had no effect at all, and FeSO$_4$brought about some improvement. However, by keeping the concentration of MgSO$_4$to be 25 mM or higher in culture broths, the antibiotic yield could be improved by more than 100%, partially due to the enhanced liberation of the intracellular antibiotic.

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Pharmacological Effect of Hawangyeonhaedoktang on Experimental Triglyceride Accumulated HepG2 Cells (실험적 중성지질 축적 HepG2세포에 미치는 황련해독탕의 약리적 효과)

  • 차재영;김대진;김석환;김영길;조영수
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.32 no.4
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    • pp.586-590
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    • 2003
  • The pharmacological effect of Korean-Chinese traditional herbal medicine, Hawangyeonhaedoktang (HT) on experimentally induced-triglyceride accumulation in cultured human hepatocyte HepG2 cells was studied. HePG2 cells were cultured in the Dulbecco's modified Eagle's (DME) medium without (Control medium) or with HT (0.5 mg/mL and 5.0 mg/mL) containing 1 mM oleate, 0.2% bovine serum albumin (BSA), and glucose 4.5 mg/mL for 6 and 24 hours in experiment I and 2 mM oleate, 0.5% BSA, and glucose 4.5 mg/mL for 6, 24 and in hours in Experiment II or 1 and 3 hours in Experiment III. Oleate [$^{14}$ C](0.5 $\mu$Ci/mL medium) added as a radioactive lipid precursor in the experiment I. In the experiment I, the intracellular triglyceride concentration was decreased remarkably during incubation for 6 and 24 hours, in a dose-dependent manner. At the same time, HT caused a decrease in the incorporation of [$^{14}$ C] oleate into intracellular triglyceride fraction and the secretion of triglyceride labeled with [$^{14}$ C] oleate into medium. In the experiment II and III compared to experiment I, the triglyceride accumulation in HepG2 cells was occurred, and HT prevented the accumulation of triglyceride during incubation for 24 and 48 hours. This result suggest that HT prevent the triglyceride accumulation in human hepatocytes by its inhibiting action on the intercellular triglyceride biosynthesis.

Inhibitory Effects of Ginsenosides on Glutamate-Induced Swelling of Cultured Astrocytes

  • Seong, Yeon-Hee;Koh, Sang-Bum;Kim, Hack-Seang
    • Journal of Ginseng Research
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    • v.24 no.3
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    • pp.138-142
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    • 2000
  • Effects of ginsenosides (Rb$_1$, Rb$_2$, Rc, Re, Rg$_1$, Rf) on L-glutamate (glutamate)-induced swelling of cultured astrocytes from rat brain cerebral cortex were studied. Following the exposure to 0.5mM glutamate for 1 hr, the intracellular water space (as measured by [$^3$H]O-methyl-D-glucose uptake) of astrocytes increased by about two-fold. Simultaneous addition of ginsenosides Rb$_2$ and Rc with glutamate reduced the astrocytic swelling in a dose-dependent manner. These ginsenosides at 0.5 mg/ml did not affect the viability of astrocytes for up to 24 hr which was determined by a colorimetric assay (MTT assay) for cellular growth and survival. These ginsenosides at 0.3 mg/ml inhibited the increase of intracellular Ca$\^$2+/ concentration ([Ca$\^$2+/]$\_$i/) induced by glutamate. These data suggest ginsenosides Rb$_2$ and Rc prevent the cell swelling of astrocytes induced by glutamate, maybe via inhibition of Ca$\^$2+/ influx.

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Effects of Carthami Tinctorius on Rat Calvarial Cell Activity and MG63 Cell mRNA Synthesis (랫드의 두개골세포 활성 및 mRNA 발현에 미치는 홍화자의 영향)

  • Kim, Tae-Woong;Hwang, Gwi-Seo
    • Journal of Society of Preventive Korean Medicine
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    • v.11 no.1
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    • pp.9-21
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    • 2007
  • This study was performed to evaluate the effect of Carthami tinctorius(HH) on osteoblast function and gene expression. The osteoblasts separated from the rat calvariae were cultivated to evaluate the cell function, and MG-63 cell was also cultivated for the test of mRNA synthesis. In this experiments, cell proliferation of rat calvarial cells was increased by HH. PKC activity, intracellular free calcium level and collgen synthesis from calvarial cells were increased by HH, but not PKA activity. And the mRNA of $PLA_2$, COX-2, and $PGE_2$ synthase from MG-63 were decreased by HH, but the mRNA of prostacyclin synthase was increased. It is concluded that HH might increase the proliferation of calvarial cell resulted from augumentation of osteoblast activity and its mRNA synthesis.

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