• Molecules and Cells
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• 제25권1호
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• pp.55-63
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• 2008

Amyotrophic lateral sclerosis (ALS) is a progressive neurodegenerative disorder characterized by the selective death of motor neurons. Mutations in the SOD1 gene are responsible for a familial form of ALS (FALS). Although many studies suggest that mutant SOD1 proteins are cytotoxic, the mechanism is not fully understood. To investigate the role of mutant SOD1 in FALS, human SOD1 genes were fused with a PEP-1 peptide in a bacterial expression vector to produce in-frame PEP-1-SOD fusion proteins (wild type and mutants). The expressed and purified PEP-1-SOD fusion proteins were efficiently transduced into neuronal cells. Neurones harboring the A4V, G93A, G85R, and D90A mutants of PEP-1-SOD were more vulnerable to oxidative stress induced by paraquat than those harboring wild-type proteins. Moreover, neurones harboring the mutant SOD proteins had lower heat shock protein (Hsp) expression levels than those harboring wild-type SOD. The effects of the transduced SOD1 fusion proteins may provide an explanation for the association of SOD1 with FALS, and Hsps could be candidate agents for the treatment of ALS.

• Molecular & Cellular Toxicology
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• 제4권2호
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• pp.106-112
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• 2008

Parkinson's disease (PD) progresses severely by a gradual loss of dopaminergic neurons in the substantia nigra (SN). Epidemiological studies showed that the incidences of PD were reduced by smoking of which the major component, nicotine might be neuroprotective. But the function of nicotine, which might suppress the incidences of PD, is still unknown. Fortunately, recently it was reported that a glial reaction and inflammatory processes might participate in a selective loss of dopaminergic neurons in the SN. The levels of tumour necrosis factor (TNF)-${\alpha}$ synthesised by astrocytes and microglia are elevated in striatum and cerebrospinal fluid (CSF) in PD. TNF-${\alpha}$ kills the cultured dopaminergic neurons through the apoptosis mechanism. TNF-${\alpha}$ release from glial cells may mediate progression of nigral degeneration in PD. Nicotine pretreatment considerably decreases microglial activation with significant reduction of TNF-${\alpha}$ mRNA expression and TNF-${\alpha}$ release induced by lipopholysaccharide (LPS) stimulation. Thus, this study was intended to explore the role of nicotine pretreatment to inhibit the expressions of TNF-${\alpha}$ mRNA in human fetal astrocytes (HFA) stimulated with IL-$1{\beta}$. The results are as follows: HFA were pretreated with 0.1, 1, and $10{\mu}g/mL$ of nicotine and then stimulated with IL-$1{\beta}$ (100 pg/mL) for 2h. The inhibitory effect of nicotine on expressions of TNF-${\alpha}$ mRNA in HFA with pretreated $0.1{\mu}g/mL$ of nicotine was first noted at 8hr, and the inhibitory effect was maximal at 12 h. The inhibitory effect at $1{\mu}g/mL$ of nicotine was inhibited maximal at 24 h. Cytotoxic effects of nicotine were noted above $10{\mu}g/mL$ of nicotine. Moreover, Nicotine at 0.1, 1 and $10{\mu}g/mL$concentrations significantly inhibited IL-$1{\beta}$-induced TF-${\kappa}B$ activation. Collectively, these results indicate that in activated HFA, nicotine may inhibit the expression of TNF-${\alpha}$ mRNA through the pathway which suppresses the NF-${\kappa}B$ activation. This study suggests that nicotine might be neuroprotective to dopaminergic neurons in the SN and reduce the incidences of PD.

• 한국잡초학회지
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• 제11권3호
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• pp.187-194
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• 1991

온도조건(溫度條件)에 따른 수도(水稻)와 잡초간(雜草間) Naproanilide의 선택성(選擇性) 기작(機作)을 구명(究明)하고자 벼, 피 올미 및 너도방동사니에서 $^{14}C$-naproanilide의 흡수(吸收), 이행(移行) 및 대사(代謝) 실험(實驗)을 수행(遂行)하였다. 1. $^{14}C$-naproanilide 흡수(吸收)도 수도(水稻)와 피보다는 감수성(感受性) 초종(草種)인 너도방동사니와 올미에서 전반적으로 많은 경향(傾向)이었으며 특히 올미에서는 $32^{\circ}C$에서 48시간 처리(處理)한 경우 흡수량(吸收量)이 배(倍) 정도 증가되었다. 2. $^{14}C$-naproanilide 이행(移行)은 수도(水稻)와 피, 너도방동사니에서 매우 적으며 거의 온도(溫度)에 영향(影響)을 받지 않았으나, 올미는 $32^{\circ}C$에서 다른 초종(草種)에 비해 이행량(移行量)도 다소 많은 경향이었다. 3. Naproanilide에 내성(耐性)을 보인 벼와 피에서 보다 감수성(感受性)을 나타낸 올미와 너도방동사니에서는 제초활성(除草活性) 대사물질(代謝物質)일 NOP와 NOPM가 Naproanilide에 비하여 현저히 많았으며 특히 올미에서는 다른 초종(草種)에 비하여 $32^{\circ}C$에서 그 경향(順向)이 현저하였다. 따라서 초종간(草種間) Naproanilide 선택성(選擇性)은 주로 활성화(活性化) 대사작용(代謝作用) 속도(速度)의 차이(差異)에 기인(基因)되며 또한 흡수량(吸收量)도 관련(關聯)되는 것으로 사료(思料)된다.

• 한국잡초학회지
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• 제17권2호
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• pp.192-198
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• 1997

벼와 피간(間)의 속간(屬間) 선택성(選擇性)을 구명(究明)하기 위하여 제초제(除草劑) cyhalofop-BE에 대(對)한 Acetyl-CoA carboxylase의 효소활성(酵素活性), 지방산(脂肪酸) 및 생합성(生合成) 단백질(蛋白質) 패턴에 미치는 영향(影響)을 시험(試驗)한 결과(結果)를 요약(要約)하면 다음과 같다. 1. ACCase을 저해하는 농도는 피가 1-2ppm 농도(濃度)의 낮은 수준(水準)이었고 벼는 10배(倍) 이상(以上)의 높은 농도(濃度)에서 저해(沮害)되었다. 2 벼 및 피의 지방산(脂肪酸) 생합성(生合成)에 관여(關與)하는 ACCase의 효소(酵素)가 저해(沮害)되어 피는 포화지방산(飽和脂肪酸)인 palmitic acid가 무처리(無處理)에 비(比)해 61%, 불포화지방산(不飽和脂肪酸)인 linoleic acid가 54%, linolenic acid는 41%가 정도(程度) 억제(抑制)되었으나 벼는 무처리(無處理)와 큰 차이(差異)를 보이지 않았다. 3. 피는 29KD와 36KD사이 및 36KD 와 45KD사이의 spot가 소멸(消滅)되어 단백질(蛋白質)패턴에 영향(影響)이 있었다.

• 한국가금학회지
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• 제16권4호
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• pp.219-232
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• 1989

본 연구는 닭에 있어서 사료성분에 대한 췌장소화효소(amylase. trypsinogen 및 chymotrypsinogen) 분비기구에 대해서 검토했다. 먼저, 췌효소분비의 단기응답실험에 유용한 새로운 췌액채취법을 개발했다. 이 방법을 이용해서 아미노산 및 glucose를 날개정맥으로 투여한 결과 phenylalanine만이 trypsinogen 및 Chymotrypsinogen이 증가되었지만 그 외의 아미노산 및 glucose에 의해서는 분비증가 효과가 없었다. Cholecystokinin(CCK)투여 에 의해 췌효소분필는 즉각적으로 높은 분비반응을 보였으며, 이 반응은 또한 농도의존성을 나타냈다. CCK투여는 chymotrypsinogen의 쪽이 amylase 및 trypsinogen보다 높은 비율로 분비되는 선택적인 분비반응을 나타냈다. 아미노산과 CCK을 공동투여하면 첨가한 아미노산의 종류에 따라 췌효소분비반응은 여러 가지 형태로 증가되었지만 glucose와의 공동투여에서는 CCK 단독투여와 비교해서 차가 없었다. Valine과 arginine을 여러 가지 농도로 CCK와 공동 투여한 결과, valnine에서는 0.5mM일때, arginin에서는 5mM일때 가장 높은 분비반응을 보였다. 위의 결과로부터 아미노산의 조합에 의한 췌효소분비반응에 대해서 검토했다. 즉, 아미노산 mixture, threonine＋phenylalanine＋isoleucine, Threonine＋phenylalanine, threonine＋isoleucine 및 phenylalanine＋isoleucine과 CCK를 공동투여 했다. 각 물질을 투여한 후 50분간 분비한 효소를 비교하면, threonine＋phenylalanine에 의한 췌효소분비반응은 아미노산 mixture에 의한 분비반응과 동일하게 높은 반응을 보였다. 이상의 결과로부터 닭에 있어서 췌장소화효소분비는 CCK와 아미노산의 사이에 협동작용이 있으며, 그 협동작용은 아미노산의 종류에 따라 선택적인 분비반응을 함으로써 장내소화가 진행된다고 본다.

• 지질공학
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• 제20권2호
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• pp.183-189
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• 2010

제3기 분지의 경계단층을 따라 발달하는 단층핵에서 단층비지 시료를 채취하여 현미경하에서 잔류입자들의 미구조 관찰과 잔류입자들을 이용한 프랙탈 차원 분석을 수행하였다. 타원형의 잔류입자들은 기질의 점토엽리(P엽리)에 평행 내지 아평행하게 형태선택배향을 이루는데 이는 풍부한 기질의 미세 입자 속에서 지속적인 단층 슬립 동안 회전에 의해 이루어진 것으로 생각된다. 잔류입자 크기 분포는 2.40-3.02 범위의 프랙탈 차원(D)을 갖는 멱 법칙을 따른다. 여기서 한 개의 시료를 제외한 모든 시료들은 Sammis et al. (1987)의 자기유사 파쇄 과정을 예측한 구속 분쇄 모델의 특정 차원 값 2.58보다 높은 값을 보이며 큰 단층 슬립과 다중단층작용을 지시한다. 아마도 비지대의 높은 차원 값은 구속 분쇄모델의 적용 범주를 지난 후부터는 파쇄 기구가 바뀌어 입자마모와 이에 부수적으로 조립입자의 선택적 파쇄가 일어남에 기인한 단층암의 비 자기유사 진화를 지시한다. 단층핵의 파쇄 진화를 통하여 후기의 입자마모 동안에 입자 파쇄가 부수적으로 일어날 수 있는 것과 마찬가지로 조기의 대량 파쇄 동안에도 부분적으로 입자마모가 수반될 수 있을 것으로 생각된다.

• Journal of Microbiology and Biotechnology
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• 제21권12호
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• pp.1345-1351
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• 2011

Vancomycin therapy failure due to the emergence of tolerance in pneumococci is increasing. The molecular mechanism of tolerance is not clear, but lytA and $pep^{27}$ are known to be involved. Our aim was to evaluate the expression of both genes in vancomycin-tolerant Streptococcus pneumoniae (VTSP) strains. Eleven VTSP strains from a total of 309 clinical isolates of S. pneumoniae from 1997 to 2006 were classified according to the criteria of Liu and Tomasz. All VTSP strains were evaluated for susceptibility according to CLSI criteria, serotype by the Quellung test, and clonality by PFGE. The expressions of lytA and $pep^{27}$ were analyzed in different growth phases by RT-PCR with and without vancomycin. Eighty-two percent of VTSP strains showed resistance to penicillin, and 100% were sensitive to vancomycin and cefotaxime. The most frequent serotypes of VTSP strains were 23F (4/11) and 6B (3/11). Clonal relationship was observed in only two strains. No significant changes were observed in $pep^{27}$ expression in the three phases of growth in VTSP strains with and without vancomycin. Interestingly, $pep^{27}$ expression in the stationary phase in the non-tolerant reference strain R6 was significantly higher. However, no significant differences in lytA expression were observed between VTSP and R6 strains during the phases of growth analyzed. The absence of changes in $pep^{27}$ expression in VTSP strains in the stationary phase may be related to their ability to tolerate high antibiotic concentrations, and thus, they survive and remain in the host under the antibiotic selective pressure reflected in therapeutic failure.

• The Korean Journal of Physiology and Pharmacology
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• 제21권3호
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• pp.309-316
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• 2017

Transient receptor potential vanilloid 3 (TRPV3) is a non-selective cation channel with modest permeability to calcium ions. It is involved in intracellular calcium signaling and is therefore important in processes such as thermal sensation, skin barrier formation, and wound healing. TRPV3 was initially proposed as a warm temperature sensor. It is activated by synthetic small-molecule chemicals and plant-derived natural compounds such as camphor and eugenol. Schisandra chinensis (Turcz.) Baill (SC) has diverse pharmacological properties including antiallergic, anti-inflammatory, and wound healing activities. It is extensively used as an oriental herbal medicine for the treatment of various diseases. In this study, we investigated whether SC fruit extracts and seed oil, as well as four compounds isolated from the fruit can activate the TRPV3 channel. By performing whole-cell patch clamp recording in HEK293T cells overexpressing TRPV3, we found that the methanolic extract of SC fruit has an agonistic effect on the TRPV3 channel. Furthermore, electrophysiological analysis revealed that ${\gamma}$-schisandrin, one of the isolated compounds, activated TRPV3 at a concentration of $30{\mu}M$. In addition, ${\gamma}$-schisandrin (${\sim}100{\mu}M$) increased cytoplasmic $Ca^{2+}$ concentrations by approximately 20% in response to TRPV3 activation. This is the first report to indicate that SC extract and ${\gamma}$-schisandrin can modulate the TRPV3 channel. This report also suggests a mechanism by which ${\gamma}$-schisandrin acts as a therapeutic agent against TRPV3-related diseases.

• The Korean Journal of Physiology and Pharmacology
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• 제4권3호
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• pp.185-195
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• 2000

Vascular diseases are significant complications of diabetes mellitus (DM), and the endothelial cells may play a pivotal role in the development of vascular disease in DM. Endothelin-1 (ET-1) released from endothelium is a potent vasoconstrictor peptide and circulating level of ET-1 is increased in a variety of disease states. The purpose of this study was to determine the changes of responsiveness to ET-1 in DM, and we experimented on the changes in the ET-1-induced contraction, levels of nitrite and lipid peroxidation, and ET-1 immunoreactivity in aorta from streptozotocin-induced DM rats. DM was induced by single injection of streptozotocin (55 mg/kg, i.p.). The immunoreactive ET-1 levels in endothelial layer of thoracic aorta were much higher in DM rats than control rats. Nitrite in tissue homogenate was decreased and plasma nitrite was increased in DM rats. Malondialdehyde (MDA) was significantly increased in DM rats and cGMP was not significantly different between control and DM rats. ET-1 produced concentration- dependent contractile responses that are significantly attenuated in DM rats compared to controls. In the presence of selective $ET_A$ receptor antagonist BQ610, the maximum contraction was decreased and the concentration ratios for BQ610 yielded $pA_2$ values of 7.3 (slope, 0.65) in control rats, whereas BQ610 had no antagonistic effect on ET-1-induced contraction in DM rats. However, pretreatment with BQ788, an $ET_B$ receptor antagonist, maximum response was decreased and the dose-response curves for ET-1 were shifted to the right in both groups and $pA_2$ values were 7.9 and 7.7 (slope, 1.05 in control and DM rats), respectively. IRL 1620 and sarafotoxin S6c, $ET_B$ agonists, induced relaxation in control rats but not in DM rats. These results indicate that endothelial cell dysfunction and enhanced immunoreactivity of ET-1 have been found in DM rat and ET-1-induced contraction was attenuated in DM rat. These attenuated responses might be at least in part caused by the alteration of $ET_A$ receptor properties (e.g. desensitization), and partly related with an alteration in intracellular mechanism for contraction to ET-1.

• The Korean Journal of Physiology and Pharmacology
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• 제13권6호
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• pp.517-526
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• 2009

The present study was attempted to investigate whether polyphenolic compounds isolated from wine, which is brewed from Rubus coreanum Miquel (PCRC), may affect the release of catecholamines (CA) from the isolated perfused adrenal medulla of the spontaneously hypertensive rats (SHRs), and to establish its mechanism of action. PCRC $(20\sim180\;{\mu}g/ml)$ perfused into an adrenal vein for 90 min relatively dose-dependently inhibited the CA secretory responses to ACh (5.32 mM), high $K^+$ (56 mM), DMPP $(100\;{\mu}M)$ and McN-A-343 $(100\;{\mu}M)$. PCRC itself did not affect basal CA secretion (data not shown). Also, in the presence of PCRC $(60\;{\mu}g/ml)$, the CA secretory responses to veratridine (a selective $Na^+$ channel activator $(10\;{\mu}M)$, Bay-K-8644 (a L-type dihydropyridine $Ca^{2+}$ channel activator, $10\;{\mu}M$), and cyclopiazonic acid (a cytoplasmic $Ca^{2+}$-ATPase inhibitor, $10\;{\mu}M$) were significantly reduced, respectively. In the simultaneous presence of PCRC $(60\;{\mu}g/ml)$ and L-NAME (an inhibitor of NO synthase, $30\;{\mu}M$), the inhibitory responses of PCRC on the CA secretion evoked by ACh, high $K^+$, DMPP, and Bay-K-8644 were considerably recovered to the extent of the corresponding control secretion compared with that of PCRC-treatment alone. The level of NO released from adrenal medulla after the treatment of PCRC $(60\;{\mu}g/ml)$ was greatly elevated compared with the corresponding basal level. Taken together, these results demonstrate that PCRC inhibits the CA secretion from the isolated perfused adrenal medulla of the SHRs evoked by stimulation of cholinergic receptors as well as by direct membrane-depolarization. It seems that this inhibitory effect of PCRC is mediated by blocking the influx of calcium and sodium into the adrenal medullary chromaffin cells of the SHRs as well as by inhibition of $Ca^{2+}$ release from the cytoplasmic calcium store at least partly through the increased NO production due to the activation of NO synthase.