• The Plant Pathology Journal
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• 제25권4호
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• pp.322-327
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• 2009

A total of 23 isolates of Botrytis cinerea causing the grey mold were collected from infected ginseng in several fields of Korea. The sensitivity to carbendazim and the mixture of carbendazim plus diethofencarb was determined through a mycelial inhibition test on PDA amended with or without fungicides. B. cinerea isolates were classified as 3 phenotypes, which were the first phenotype resistant to both of carbendazim and the mixture ($Car^RMix^R$), the second one resistant to carbendazim and sensitive to the mixture ($Car^RMix^S$), and the last one sensitive to both of them ($Car^RMix^S$). Carbendazim resistance correlated with a single mutation $\beta$-tubulin gene of B. cinerea amplified with primer pair tubkjhL and tubkjhR causing a change of glutamate to alanine at amino acid position 198. Furthermore, the substitution of valine for glutamate led the resistance to carbendazim and the mixture at the same position of amino acid. PCR-restriction fragment length polymorphism (PCR-RFLP) analysis using the restriction endonuclease, Tsp451 and BstUI allowed differentiation of the PCR fragment of $\beta$-tubulin gene of $Car^SMix^S$ isolates from that of $Car^RMix^R$ and $Car^RMix^S$ isolates. This method will aid in a fast detection of resistance of carbendazim and the mixture of carbendazim plus diethofencarb in B. cinerea in ginseng field.

• Applied Biological Chemistry
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• 제45권4호
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• pp.190-194
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• 2002

Bacillus subtilis의 glutamyl-tRNA synthetase(GluRS)는 대장균에서 발현될 때 숙주세포의 $tRNA_1^{Gln}$에 glutamate를 잘못 아실화하여 독성을 나타내는 것으로 추정되고 있다. 이러한 B. subtilis GluRS를 대장균에서 과발현 시키기 위하여 B. subtilis 168 균주의 chromosomal DNA에서 GluRS의 유전자(gltX)를 PCR을 이용하여 증폭하고 T7 promoter에 의해 발현이 조절되는 pET11a expression vector에 클로닝하였다. 이 재조합된 pEBER plasmid DNA로 T7 RNA polymerase를 갖는 대장균 NovaBlue(DE3)에 형질전환하였다. 형질전환된 대장균에 IPTG를 처리하여 과량 생성된 GluRS 단백질은 ammonium sulfate 분별침전 후 EPLC를 이용한 Source Q column anion exchange chromatography, Superdex 200 column gel filtration, Mono Q column anion exchange chromatography로 정제하였다. 정제된 B. subtilis의 GluRS 분자량은 약 55 kDa이었으며 효소의 활성도는 조효소액에 비해 18배로 증가하였다.

• The Korean Journal of Physiology and Pharmacology
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• 제7권6호
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• pp.295-301
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• 2003

Striatum plays a crucial role in the movement control and habitual learning. It receives an information from wide area of cerebral cortex as well as an extensive serotonergic (5-hydroxytryptamine, 5-HT) input from raphe nuclei. In the present study, the effects of 5-HT to modulate synaptic transmission were studied in the rat corticostriatal brain slice using in vitro extracellular recording technique. Synaptic responses were evoked by stimulation of cortical glutamatergic inputs on the corpus callosum and recorded in the dorsal striatum. 5-HT reversibly inhibited coticostriatal glutamatergic synaptic transmission in a dose-dependent fashion (5, 10, 50, and $10{\mu}M$), maximally reducing in the corticostriatal population spike (PS) amplitude to $40.1{\pm}5.0$% at a concentration of $50{\mu}M$ 5-HT. PSs mediated by non-NMDA glutamate receptors, which were isolated by bath application of the NMDA receptor antagonist, d,l-2-amino-5-phospohonovaleric acid (AP-V), were decreased by application of $50{\mu}M$ 5-HT. However, PSs mediated by NMDA receptors, that were activated by application of zero $Mg^{2+}$ aCSF, were not significantly affected by $50{\mu}M$ 5-HT. To test whether the corticostriatal synaptic inhibitions by 5-HT might involve a change in the probability of neurotransmitter release from presynaptic nerve terminals, we measured the paired-pulse ratio (PPR) evoked by 2 identical pulses (50 ms interpulse interval), and found that PPR was increased ($33.4{\pm}5.2$%) by 5-HT, reflecting decreased neurotransmitter releasing probability. These results suggest that 5-HT may decrease neurotransmitter release probability of glutamatergic corticostriatal synapse and may be able to selectively decrease non-NMDA glutamate receptor-mediated synaptic transmission.

• 한국수소및신에너지학회논문집
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• 제22권4호
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• pp.451-457
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• 2011

Purple non-sulfur (PNS) bacterium $Rhodobacter$ $sphaeroides$ KD131 was studied with the aim of achieving maximum hydrogen production using various carbon and nitrogen sources at different pH conditions. Cells grew well and produced hydrogen using $(NH_4){_2}SO_4$ or glutamate as a nitrogen source in combination with a carbon substrate, succinate or malate. During 48h of photo-heterotrophic fermentation under 110$W/m^2$ illumination using a halogen lamp at $30^{\circ}C$, 67% of 30mM succinate added was degraded and the hydrogen yield was estimated as 3.29mol $H^2$/mol-succinate. However, less than 30% of formate was consumed and hydrogen was not produced due to a lack of genes coding for the formate-hydrogen lyase complex of strain KD131. Initial cell concentrations of more than 0.6g dry cell weight/L-culture broth were not favorable for hydrogen evolution by cell aggregation, thus leading to substrate and light unavailability. In a modified Sistrom's medium containing 30mM succinate with a carbon to nitrogen ratio of 12.85 (w/w), glutamate produced 1.40-fold more hydrogen compared to ammonium sulfate during the first 48h. However, ammonium sulfate was 1.78-fold more effective for extended cultivation of 96h. An initial pH range from 6.0 to 9.0 influenced cell growth and hydrogen production, and maintenance of pH 7.5 during photofermentation led to the increased hydrogen yield.

• 대한임상검사과학회지
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• 제43권3호
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• pp.113-119
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• 2011

This study was designed to determine the effects of larval extracts from Protaetia brevitarsis seulensis shade dried larva from Korea and China on liver tissues of hepatic injury groups. The experimental groups are divided into five groups; Normal, carbon tetrachloride single injection ($CCl_4$), Pb-CHI ($CCl_4$ + P. brevitarsis shade dried larva from China), Pb-KOR ($CCl_4$ + P. brevitarsis shade dried larva from Korea) and SIL ($CCl_4$ + 0.35% silymarin) groups. Sprague Dawley rats were oral injected with $CCl_4$ at a dose l mg/kg (20% in corn oil) for induction of liver damage for 4 weeks. Each experimental group was fed with a dose 50 mg/kg of larval extracts based on medicinal preparations from 3 weeks to 4 weeks after $CCl_4$ treatment. At the end of 4 weeks, we evaluated the serum levels of glutamate-oxaloacetate transaminase (GOT), glutamate-pyruvate transferase (GPT), alkaline phosphatase (ALP) and blood urea nitrogen (BUN) in serum and the cytokine levels of tumor necrosis factor-${\alpha}$) (TNF-${\alpha}$) and transforming growth factor-${\beta}$ (TGF-${\beta}$) in the cells isolated from spleen and liver. The histological analysis was also conducted. The $CCl_4$ injection reduced body weight, induced congestion of middle lobe and hepatocytic degeneration, resulting in disintegration of hepatic cords, and increased biochemical markers of blood related to hepatic injury. On the other hand, the Pb-CHI and Pb-KOR group decreased the levels of biochemical markers in blood and cytokine levels in spleen and liver. Especially, the Pb-KOR group facilitated the recovery of biochemical values of blood related to hepatic injury, hepatic lesions and fibrosis. Taken together, larval extract from P. brevitarsis might prevent acute hepatotoxicity and enhance the recovery of liver fibrosis and cirrhosis induced by $CCl_4$, and the ingredients could be a promising candidate for the prevention and treatment of hepatic disorders.

• 생명과학회지
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• 제16권7호
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• pp.1123-1126
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• 2006

소나무 잔나비버섯은 약용으로 이용하고자 구성아미노산, 유리아미노산, 비타민, 무기물의 성을 분석 검토한 결과 건조버섯 100 g당 총아미노산은 2901.5 mg, 유리아미노산 68.67 mg, 비타민 276.91 mg, 무기물은 347.37 mg이었다. 총 아미노산의 구성아미노산은 18종의 아미노산으로 구성되었으며, glutamic acid가 457 mg로 가장 많이 함유되었고, threonine, glycine, aspartic acid, valine등의 순으로 각각 279 mg, 241 mg, 233 mg, 233 mg의 함량이었다. 유리아미노산 중에서는 glutamine이 17.61 mg으로 가장 많이 함유되어 있고, arginine, tryptophan, threonine, glutamic acid, isoleucine, phenylalanine, tyrosine, valine이 각각 6.51 mg, 5.65 mg, 4.38 mg, 4.02 mg, 3.89 mg, 2.69 mg, 1.51 mg, 1.87 mg등의 순으로 구성되었다. 소나무잔나비 버섯의 무기물 구성 중, 성분 P가 77.57 mg로 가장 많이 함유되었고 다음으로 Mg, Fe, Ca, Na, Zn,등의 순이었다. 풍부한 아미노산 및 무기물 함량과 더불어 소나무잔나비 버섯의 비타민 성분 중, vitamin E는 275.76 mg으로 노화방지에 효과가 높을 것으로 사료되고, 추후 기능성 식품으로 이용적 가치가 높을 것으로 판단된다.

• 한국어병학회지
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• 제23권3호
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• pp.409-420
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• 2010

Effects of various concentrations of garlic powder and garlic extract in the diets on growth, serum chemistry and immune response of olive flounder were determined. Thirty-five juvenile fish averaging 5.1 g were randomly distributed into 21 of 180 L flow-through tanks. Seven experimental diets with various concentrations of garlic powder (GP) and garlic extract (GE) were prepared in triplicate: GP-0 without garlic supplementation, GP-0.5, GP-1, GP-2, GP-3 and GP-5 diets containing garlic powder at the concentrations of 0.5, 1, 2, 3 and 5%, respectively at the expense of wheat flour and finally, GE-0.4 diet containing 0.4% garlic extract were prepared. At the end of the 8-week feeding trial, serum chemistry of fish was measured. In addition, twenty fish from each tank were artificially infected with E. tarda for the following 96 h to monitor cumulative mortality. Weight gain of fish fed GP-0 diet was higher than that of fish fed GP-1, GP-2, GP-3 and GP-5 diets. No difference in serum criteria (total protein, glucose, glutamate oxaloacetate transaminase, cholesterol and triglyceride levels) of olive flounder was found among the experimental diets except for glutamate pyruvate transaminase. Lysozyme activity of fish fed GP-0, GP-1, GP-3 and GE-0.4 diets was higher than that of fish fed GP-5 diet. The highest cumulative mortality was 93.3% in fish fed GP-0 diet at 96 h after E. tarda infection, followed by GP-3, GP-1, GP-5, GP-2, GP-0.5 and GE-0.4 diets. In considering these results, dietary inclusion of garlic powder and garlic extract has no distinctive positive effect on improvement in growth, serum chemistry and immune response of olive flounder in this experimental conditions, therefore, its application should be carefully considered.

• Journal of Korean Neurosurgical Society
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• 제29권8호
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• pp.1008-1018
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• 2000

Objective : Glutamate induced excitotoxicity is one of the leading causes of cell death under pathologic condition. However, there is controversy whether excitotoxicity may also participate in the neuronal death under low intensity insult such as simple hypoxia or hypoglycemia. To investigate the role of NMDA receptor in low intensity insult, we chose anoxia as the method of injury and used organotypically cultured hippocampal slice as the material of experiment. Materials & Methods : The hippocampal slices cultured for 2-3 weeks were exposed to 60 minutes of complete oxygen deprivation(anoxia). Neuronal death was assessed with Sytox stain. Corrected optical density of fluorescence in gray scale, used as cellular death indicator, was obtained from pictures taken at 24 and 48 hours following the insult. The well-known in vivo phenomenon of regional difference in susceptibility of hippocampal sub-fields to ischemic insult was reproduced in HOSC(hippocampal organotypic slice culture) by complete oxygen deprivation injury. Results : $CA_1$ was the most vulnerable to complete oxygen deprivation in hippocampus while $CA_3$ was resistant. Oxygen deprivation for 10 and 20 minutes with glucose(6.5g/l) present was insufficient to induce neuronal death in the cultured hippocampal slice. However, after 30 minutes exposure under anoxic condition, neuronal death was able to be detected in the center of $CA_1$ area. The intensity and area of fluorescence indicating cell death correlated with the duration of oxygen deprivation. NMDA receptor and non-NMDA receptor blocking with MK-801(30 & $60{\mu}M$) and CNQX($100{\mu}M$) did not provide cellular protection to HOSC against damage induced by oxygen deprivation, but increased intracellular calcium buffering capacity with BAPTA-AM($10{\mu}M$) was effective in preventing neuronal death (p=0.01, Student's t-test). Cycloheximide($1{\mu}g/ml$, $10{\mu}g/ml$) provided no protection to HOSC against insult of complete oxygen deprivation for 60 minutes and combined therapy of MK-801(30 & $60{\mu}M$) and cycloheximide(1 & $10{\mu}g/ml$) was also ineffective in preventing neuronal death. Conclusion : The results of this study show that the another mechanism not associated with glutamate receptor(NMDA & non NMDA) may play major role in cell death mechanisms induced by complete oxygen deprivation and increased intracellular calcium during anoxia may participate in the neuronal death mechanism of oxygen deprivation. Further investigation of the calcium entry channel activated during oxygen deprivation is necessary to understand the neuronal death of anoxia.

• Asian-Australasian Journal of Animal Sciences
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• 제20권5호
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• pp.733-741
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• 2007

The effects of intra-duodenal infusion of methionine (Met), lysine (Lys) and leucine (Leu) on dry matter intake (DMI), the concentrations of insulin-like growth factor I (IGF-I), growth hormone (GH) and insulin in plasma, and liver IGF-I mRNA level were investigated in two experiments for Liuyang Black growing wether goats. In Experiment 1, three goats ($10.0{\pm}0.1$ kg) were fitted with ruminal, proximal duodenal and terminal ileal fistulaes to determine the infusion amounts of Met, Lys and Leu at the duodenum according to essential amino acid flows into the duodenum and their apparent digestibility. The infusion amounts were 0.77 g/d, 0.91 g/d and 0.58 g/d respectively. In Experiment 2, 4 groups of goats (($10.0{\pm}0.2$ kg) for each group, were cannulated at the duodenum, and were infused with a mixture of Met, Lys and Leu (Control), or mixtures with 21% Met, Lys or Leu replaced with glutamate respectively on a nitrogenous basis. The replacement of 21% Met, Lys or Leu with glutamate did not affect intakes of maize stover, concentrate or both (p>0.05) when compared with the control. The replacement of 21% Met or Lys significantly (p<0.05) reduced plasma GH, insulin and IGF-I concentrations and liver IGF-I mRNA level. The replacement of 21% Leu with glutamate reduced (p<0.05) plasma IGF-I concentration only, but not plasma insulin and GH, as well as liver IGF-I mRNA level (p>0.05). The close relationships between supplying Met and Lys in the lumen of the duodenum and plasma IGF-I, GH and insulin concentrations, as well as liver IGF-I mRNA level in this study indicate that the effects of the limiting amino acids on nutrition of animals are likely intermediated via their effects on these hormones, and these hormone profiles could be used as intermediate markers for the limiting order of amino acids.

• The Korean Journal of Physiology and Pharmacology
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• 제1권6호
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• pp.673-679
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• 1997

It has been generally accepted that glutamate mediates the ischemic brain damage, excitotoxicity, and induces release of neurotransmitters, including norepinephrine(NE), in ischemic milieu. In the present study, the role of nitric oxide(NO) in the ischemia-induced $[^3H]norepinephrine([^3H]NE)$ release from cortex slices of the rat was examined. Ischemia, deprivation of oxygen and glucose from $Mg^{2+}-free$ artificial cerebrospinal fluid, induced significant release of $[^3H]NE$ from cortex slices. This ischemia-induced $[^3H]NE$ release was significantly attenuated by glutamatergic neurotransmission modifiers. $N^G-nitro-L-arginine$ methyl ester(L-NAME), $N^G-monomethyl-L-arginine$ (L-NMMA) or 7-nitroindazole, nitric oxide synthase inhibitors attenuated the ischemia-evoked $[^3H]NE$ release. Hemoglobin, a NO chelator, and 5, 5- dimethyl-L-pyrroline-N-oxide(DMPO), an electron spin trap, inhibited $[^3H]NE$ release dose-dependently. Ischemia-evoked $[^3H]NE$ release was inhibited by methylene blue, a soluble guanylate cyclase inhibitor, and potentiated by 8-bromo-cGMP, a cell permeable cGMP analog, zaprinast, a cGMP phosphodiesterase inhibitor, and S-nitroso-N-acetylpenicillamine (SNAP), a nitric oxide generator. These results suggest that the ischemia-evoked $[^3H]NE$ release is mediated by NMDA receptors, and activation of NO system is involved.