• Title/Summary/Keyword: cytoplasmic effect

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Effect of Extracellular Potassium on Delayed Rectifier Potassium Channel Proteins of KCNQ3 and KCNQ5 in Familial Hypokalemic Periodic Paralysis (가족성 저칼륨성 주기성 마비에서 세포외 칼륨농도가 지연성 정류형 채널을 형성하는 KCNQ3와 KCNQ5 단백질에 미치는 효과)

  • Kim, Sung-Jo;Kim, Dong-Hyun;Kim, June-Bum
    • Journal of Life Science
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    • v.19 no.10
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    • pp.1484-1488
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    • 2009
  • Familial hypokalemic periodic paralysis (HOKPP) is an autosomal dominant muscle disorder characterized by episodic attacks of muscle weakness with concomitant hypokalemia. Mutations in either a calcium channel gene (CACNA1S) or a sodium channel gene (SCN4A) have been shown to be responsible for this disease. The combination of sarcolemmal depolarization and hypokalemia has been attributed to abnormalities of the potassium conductance governing the resting membrane potential. To understand the pathophysiology of this disorder, we examined both mRNA and protein levels of delayed rectifier potassium channel genes, KCNQ3 and KCNQ5, in skeletal muscle fibers biopsied from patients with HOKOur results showed an increase in the cytoplasmic level of KCNQ3 protein in patients' cells exposed to 50 mM external concentration of potassium. However, mRNA levels of both channel genes did not show significant change in the same condition. Our results suggest that long term exposure of skeletal muscle cells in HOKPP patients to high extracellular potassium alters the KCNQ3 localization, which could possibly hinder the normal function of this channel protein. These findings may provide an important clue to understanding the molecular mechanism of familial hypokalemic periodic paralysis.

Inheritance of Tetradifon Resistance in Two-spotted Spider Mite (Acari: Tetranychidae) and Its Cross Resistance (Tetradifon에 대한 점박이응애(Tetranychus urticae Koch) 저항성의 유전양식과 교차저항성)

  • 박정규;이상계;최병렬;유재기;이정운
    • Korean journal of applied entomology
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    • v.35 no.3
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    • pp.260-265
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    • 1996
  • A field colony of the two-spotted spider mite, Tetranychus urticae Koch, was selected with tetradifon for 4 mousing whole-plant residual method. This strain showed 371-fold resistance to tetradifon. The log dosehatchability lines of $F_{1}$ eggs(RS cross; Td5female$\times$Smale, and SR cross; Sfemale$\times$Td5male) were closer to the line of the resistant colony than to that of the susceptible. These differences could be due to cytoplasmic inheritance or maternal effect. The estimate of dominance index (D) for the $F_{1}$ eggs of RS cross was 0.998 and that for $F_{1}$ eggs of SR cross was 0.262. This indicates that tetradifon resistance is completely dominant in RS cross and incompletely dominant in SR cross. Td5 strain exhibited high levels of resistance to clofentezine, benzoximate, and chlorfencon, and no cross resistance to fenazaquin, pyridaben, flufenoxuron, tebufenpyrad, and fenothiocarb.

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Functional Defect of the Fas Mutants Detected in Gastric Cancers (위암에서 발견된 돌연변이형 Fas 단백의 기능적 결함)

  • Park Won Sang;Cho Young Gu;Kim Chang Jae;Park Cho Hyun;Kim Young Sil;Kim Su Young;Nam Suk Woo;Lee Sug Hyung;Yoo Nam Jin;Lee Jung Young
    • Journal of Gastric Cancer
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    • v.3 no.4
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    • pp.186-190
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    • 2003
  • Purpose: The balance between cell proliferation and apoptosis is crucial for homeostatic maintenance in a cell population. Decreased apoptosis or uncontrolled proliferation can lead to cancer. The Fas receptor signal through a cytoplasmic death domain is very important in the apoptotic pathway. To identify the effect of the death domain of the Fas gene in the development and/or progression of gastric cancer, we examined the apoptotic potential of five known Fas mutants detected in gastric cancers. Materials and Methods: A wild-type Fas gene was cloned with cDNA from normal liver tissue and full length Fas was sequenced. Mutants of the gene were generated with sitedirected mutagenesis by using the wild-type gene and specific primers. Wild- and mutant-type genes were transfected to HEK293 cells. Forty-eight hours after transfection the cells were stained with DAPI and cell death was counted under fluorescent microscopy. Results: In wild-type Fas-transfected cells, the percentage of apoptotic cells was $85.9\pm3.6\%$, and significant cell death and classic morphologic signs of apoptosis were observed. However, the percentages of apoptotic cells transfected with N239D, E240G, D244V, and R263H of tumor-derived mutant Fas were $29.5\pm2.08\%,\;28.5\pm3.34\%,\;25.225\pm2.06\%,\;and\;36.625\pm4.49\%$, respectively. Conclusion: These results suggest that inactivation of Fas caused by mutations in the death domain of the Fas gene may be one of the possible escape mechanisms against Fas-mediated apoptosis and that inactivating mutation of the Fas may contribute to the development or progression of gastric cancers.

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The Effects of Gobonyangjeonbang Administration on Antioxidative of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) induced Rats. (고본양정방 투여가 TCDD 유도 랫드의 항산화 변화에 미치는 영향)

  • Yang, Dong Hyun;OH, Ji Hye;Park, Un kyu;Cho, Chung Sik;Hwang, Seock Yeon
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.21 no.10
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    • pp.231-239
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    • 2020
  • The purpose of this study was to evaluate the effects of gobonyangjeonbang (GYB) on the endocrine function and the antioxidant efficacy of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD)-induced oxidative stress in rats. In 2017, to evaluate the efficacy of GYB on oxidative stress, 35 male SD rats were divided into five groups and tested. The normal control group was administered saline as a vehicle, while the TCDD-alone group was administered TCDD (2 ㎍/kg per week) intraperitoneally and with physiological saline, and the test group was administered GYB orally by dividing it into three concentrations (75, 150, and 300 mg/kg) for six weeks. Bodyweight decreased significantly after six weeks of TCDD exposure, when compared to rats in the NC group (p<0.001). However, weight loss from TCDD was significantly protected by administration of GYB at 300 mg/kg (p<0.01). The rat liver induced by TCDD showed cytoplasmic vacuole degeneration, and the hepatic sinusoid and weight increased. As a result of measuring MDA and SOD, both items tended to decrease under TCDD administration. On the other hand, there was no change due to GYB administration, and significance was observed in the GYB 300 mg/kg group compared to the NC group in the SOD result (p<0.05). These findings demonstrated that GYB may have a protective effect against TCDD-induced liver toxicity in rats.

Ultrastructural Study on the Cerebellar Purkinje Cell of the Head-Irradiated Rat (과량의 방사선 국소조사가 흰쥐 소뇌 Purkinje세포의 미세구조에 미치는 영향)

  • Ahn, E-Tay;Yoon, Kyoo-Tae;Yang, Nam-Gil;Ko, Jeong-Sik;Park, Kyung-Ho;Kim, Jin-Gook
    • Applied Microscopy
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    • v.24 no.2
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    • pp.48-62
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    • 1994
  • The acute irradiation effect on rat Purkinje cell was carried out. Anesthetized rats, weighing 200-250g each, were exposed their heads to the linear accelerator (ML-4MV) with the doses of 3,000 rads or 6,000 rads respectively. Irradiated rats were sacrificed by perfusion fixation under anesthesia, six hours, two days and six days following the irradiations. Rats were perfused with the fixative of 1% glutaraldehyde-1% paraformaldehyde solution (pH 7.4). Small pieces of cerebellar cortices were taken out. Tissue blocks were washed out, and were refixed in the 2% osmium tetroxide solution. After dehydration, tissues were embedded in the araldite mixture. Ultrathin sections stained with uranyl acetate-lead citrate solution, were examined with an electron microscope. The results observed were as follow; 1. Many dark Purkinje cells exhibited most severe cellular alterations on 6 hours. But after the 2 or 6 days, the cells exhibited only some alterations of cytoplasmic organelles. 2. Many granular and agranular endoplasmic reticula exhibited the fusion of cisterns. These reticular alterations were most severe on 6 hours following irradiation. But the alterations were hardly found on 6 days. 3. In the Golgi region, alterations including the adhesion of lamelliform cisterns, enlarged saccules, and increased number of vesicles, etc, were seen on 6 hours. But the Golgi complexes were almost recovered on 6 days. 4. Lysosomes were abundant on 6 hours or 2 days, but some residual bodies were found on 6 days. 5. Mitochondrial changes were also most severe at on hours, and they were recovered thereafter. From the results, it was concluded that the cerebellar Purkinje cells reacted to the high doses of irradiation by hyperactive protein synthesis, autolytic activities and energy metabolism. The reaction was most active in the early stage. It implies that motor-control function of Purkinje cells are severely disturbed in the early stage of irradiation.

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Inhibitory Effects of Self-Fermented Pine Needle Extract on Catecholamine Release in the Rat Adrenal Medulla

  • Choi, Mee-Sung;Seo, Young-Hwan;Cheong, Hyeon-Sook;Lim, Dong-Yoon
    • Natural Product Sciences
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    • v.19 no.1
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    • pp.36-48
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    • 2013
  • The aim of the present study was to investigate the effects of several fractions obtained from methylene chloride ($CH_2Cl_2$) extract of self-fermented pine needle (SFPNE) on the acetylcholine (ACh)-evoked CA release from the isolated perfused model of the rat adrenal medulla and to establish the mechanism of the most active fraction (Fr.)-induced inhibitory action on the CA release. We obtained 6 fractions from $CH_2Cl_2$ extract of self-fermented pine needle. For the ACh (5.32 mM)-evoked CA release, the following rank order of inhibitory potency was obtained: Fr.4-5 > Fr.8-11 ${\gg}$ Fr.3 > Fr.6 = Fr.7 > Fr.1-2. Fr. 4 - 5 (60 ${\mu}g/mL$) perfused into an adrenal vein for 90 min produced relatively time-dependent inhibition of the CA secretory responses to ACh (5.32 mM), DMPP (100 ${\mu}M$), McN-A-343 (100 ${\mu}M$) and high $K^+$ (56 mM). Fr. 4 - 5 itself did not affect basal CA secretion. Also, in the presence of Fr. 4 - 5 (60 ${\mu}g/mL$), the CA secretory responses to angiotensin II (AngII, 0.1 ${\mu}M$), veratridine (50 ${\mu}M$), Bay-K-8644 (10 ${\mu}M$), and cyclopiazonic acid (10 ${\mu}M$) were significantly reduced, respectively. In the simultaneous presence of Fr. 4 - 5 (60 ${\mu}g/mL$) and L-NAME (30 ${\mu}M$), the inhibitory responses of Fr. 4 - 5 on the CA secretion evoked by ACh, DMPP, high $K^+$, AngII, Bay-K-8644 and veratridine were considerably recovered to the extent of the corresponding control secretion compared with that of Fr. 4 - 5-treatment alone. The level of NO released from adrenal medulla after the treatment of Fr. 4 - 5 (60 ${\mu}g/mL$) was greatly elevated compared with the basal level. Taken together, these results demonstrate that Fr. 4 - 5 inhibits the CA secretion from the isolated perfused rat adrenal medulla evoked by stimulation of cholinergic receptors as well as by direct membrane-depolarization. It seems that this inhibitory effect of Fr. 4 - 5 is mediated by blocking the influx of $Ca^{2+}$ and $Na^+$ into the adrenomedullary chromaffin cells as well as by inhibition of $Ca^{2+}$ release from the cytoplasmic calcium store, which is evoked at least partly through the increased NO production due to the activation of NO synthase. Based on these results, it is also thought that Fr. 4 - 5 isolated from $CH_2Cl_2$ extract of pine needle may contain beneficial antihypertensive components to prevent or treat hypertension.

Growth Inhibition of Helicobacter pylorio by Reynoutria elliptica Migo. (호장근에 의한 Helicobacter pylori의 생육 저해)

  • Lee, In-Seon;Im, Hyo-Gwon;Lee, Syng-Ook
    • Korean Journal of Food Science and Technology
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    • v.35 no.6
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    • pp.1182-1187
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    • 2003
  • This study was performed to evaluate the potentiality of Reynoutria elliptica Migo., being used as a folk remedy and a herb medicine for urethritis, cystitis, etc., on growth inhibition of Helicobacter pylori which is known as the ulcerogenic pathogen. The minimum inhibitory concentration (MIC) value of methanol extract from Reynoutria elliptica Migo, was determined to be 120 ppm for H. pylori and urease activity derived from H. pylori was inhibited over 80% by the extract at 2 mg/mL in urea broth. Among various solvent fraction of the methanol extract, the hexane fraction showed a significant inhibitory effect on the growth of H. pylori reducing both its growth and urease activity. Scanning and transmission electron micrographs of H. pylori treated with the methanol extract at 2 mg/mL for 3 hr showed that the cell walls and membranes were disrupted so that the cytoplasmic components were leaked from the body. These results suggest that Reynoutria elliptica Migo. possesses a therapeutic potential on the gastric disease caused by H. pylori.

Antibacterial activity from medicinal plant extracts on the Staphylococcus aureus (수종 한약재 추출물의 Staphylococcus aureus에 대한 항균활성)

  • Park, Chun-Geon;Bang, Kyong-Hwan;Lee, Seung-Eun;Cha, Moon-Seok;Sung, Jung-Sook;Park, Hee-Woon;Seong, Nak-Sul
    • Korean Journal of Medicinal Crop Science
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    • v.9 no.4
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    • pp.251-258
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    • 2001
  • Medicinal plant extracts including Rubus coreanus, Sanguisorba officinalis, Eriobotrya japonica, Prunus mume, Crataegus pinnatifida, Rosa leavaigate Prunus persica, Prunus japonica var. nakaii and Spiraea blumei were prepared for the test of antibacterial activity. Tryptic soy broth (TSB) containing $0{\sim}10mg/ml$ of medicinal plant extracts was inoculated with $10^6$ cells/ml of Staphylococcus aureus and incubated at $35^{\circ}C$ for 24 hours. The plate counting method and clear zone test were used to test inhibitory effect of the extracts. Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) was derived from the survival curves of S. aureus. The order of antibacterial activities of medicinal plant extracts on the S. aureus was Rubus coreanus > Sanguisorba officinalis > Eriobotrya japonica > Prunus mume > Crataegus pinnatfida. Minimum inhibitory concentration of Sanguisorba ofEcmalis on the Staphylococcus aureus was 2.5mg/ml and minimum bactericidal concentration of Rubus coreanus was 1.0%. Inhibition zone of Rubus coreanus, Sanguisorba officinalis, Eriobotrya japonica, Prunus mume, and Crataegus pinnatifida was 16.5mm, 14.3mm 11.0mm, 14.0mm and 12.7mm, respectively. The morphology of S. aureus cells treated with medicinal plant extracts showed damage of cell wall and cytoplasmic membrane. Severely damaged cells of S. aureus lost electron dense material and cytoplasm. This result suggests that medicinal plant extracts can be used as an effective natural antibacterial agent in food.

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Effect of Garlic Oil (diallyl disulfide)/ Vitamin A( retinol acetate on Heat Shock Protein Induction in Cadmium Treated Rats. (마늘유(diallyl disulfide)와 비타민 A(retinol acetate)가 카드뮴 투여 랫드에 미치는영향)

  • 김판기
    • Journal of Food Hygiene and Safety
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    • v.13 no.2
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    • pp.171-187
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    • 1998
  • Garlic occupies a special position among the many foods of vegetable origin because it is the sole food for Koreans during the their lives. And vitamin A has been ingested by forms of food or additives. Cadmium has been described as one of the most dangerous trace elements in the food and environment of man and livestocks. Since the de novo synthesis of stress proteins can be detected early after exposure to some agents, analysis of cadmium-induced changes in gene expression , ie. alterations in patterns of protein synthesis, may be useful to develop as biomarkers of exposure and damage for food hygiene. He acute and chronic combine effects of cadmium (Cd, CdCl2 20mg/kg), garlic oil(Dds: diallyl disulfide 50mg/kg, 3 times a week) and vitamin A(Ra: retinol acetate 50,000 IU/kg, 3 times a week) on Wistar male rats were evaluated concerning cadmium contents, tissues enzyme activity, HSP expression histopathological and electron microscopical examinations. The results of the study are as follows ; 1. Less cadmium was absorbed through the digestive tracts, but the ratio of contents in tissue were not changed by the simultaneous adminstration of diallyl disufide or retinol acetate. 2. ALT(alanine aminotransferase) , AST(aspartate aminotransferase), glucose, BUN (blood urea nitrogen), creatinine, the key indices of the clinical changes in hepatic and renal function were significantly hanged by the cadmium treatment after 1 week in liver, after 4 weeks in kidney. 3. Histopathological changes in cadmium treated rats were appeared at 8 weeks age treatment in kidneys. Homogenous eosinophilic material was accumulated in cortical and collecting tubular lumens at 16 weeks. Degenerated or necrotized tubular cells were observed in cortex and medulla. Degenerated seminiferous tubules and homogeneous eosinophilic material was seen in interstitial tissue of rat treated with cadmium for 16 weeks. Calcium deposits were seen in degenerated seminiferous tubules and the tubules showed severe calcification of rat treated with cadmium for 16 weeks. Electron microscope changes in kidney were observed in rats treated with CdCl2 20 mg/kg. Proximal convoluted tubule cells showed selling of cytoplasm and narrow lumen. Capillary endothelial cells showed cytoplasmic vacuoles and swelling. Degenerated epithelial cells were accumulated in tubular lumen of kidney. 4. Enhanced synthesis of 70 KDa relateve molecular mass proteins were detected in 2 hours after cadmium, exposure, with maximum activity occurring at 8~48 hours. Induction of HSP 70 was evident at proximal tubules and glomeruli in kidney. Testicular cells produced enough HSP to be detected normally. From the above results, it could be concluded that HSP70 induction by the cadmium treatment was a rapid reaction to indicated the exposure of xenobiotics, and retinol acetate reduced the cadmium induced nephrotoxicity.

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Pituitary Adenylate Cyclase-activating Polypeptide (PACAP) Treatment during Pre-maturation Increases the Maturation of Porcine Oocytes Derived from Small Follicles

  • Park, Kyu-Mi;So, Kyoung-Ha;Hyun, Sang-Hwan
    • Journal of Embryo Transfer
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    • v.33 no.1
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    • pp.1-11
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    • 2018
  • Cellular cyclic adenosine-3' 5'-monophosphate (cAMP) modulator is known as meiotic inhibitor and can delays spontaneous maturation in IVM experiment. Among many cAMP modulators, the role of Pituitary adenylate cyclase activating polypeptide (PACAP) on IVM isn't known. The purpose of this study is to improve the maturation of oocytes derived from follicles ${\leq}3mm$ in diameter through PACAP as meiotic inhibitor during pre-in vitro maturation (pre-IVM). First, we checked PACAP and its receptors in cumulus cells and, to establish the optimal phase and concentration of PACAP for pre-IVM, we conducted chromatin configuration assessments. As a result, the rate of GV (Germinal Vesicle) according to duration of pre-IVM was significantly decreased 12 h and 18 h after IVM (87.1 and 84.1%, respectively) compared to 0 h (99.4%). When COC was cultured for 18 h, the GV rate in the $1{\mu}M$ of PACAP treatment group (82.1%) was significantly higher than any other PACAP treatment groups (60.5, 64.1, 74.4 and 69.9 %, respectively). So, we divided into four groups as follows; MF (the conventional IVM group, obtained from follicle from 3 to 6 mm in diameter), SF (the conventional IVM group, obtained from follicle ${\leq}3mm$ in diameter), Pre-SF(-)PACAP (IVM group including 18 h pre-IVM without $1{\mu}M$ of PACAP, obtained from follicle ${\leq}3mm$ in diameter) and Pre-SF(+)PACAP (IVM group including 18 h pre-IVM with $1{\mu}M$ of PACAP, obtained from follicle ${\leq}3mm$ in diameter). To examine the effect of PACAP during pre-IVM, we investigated analysis of nuclear maturation, intracellular glutathione (GSH) and reactive oxygen species (ROS) levels. In cumulus cells, PACAP receptors, ADCYAP1R1 and VIPR1 were detected but were not detected in oocytes. After IVM, the Pre-SF(+)PACAP had the highest Metaphase II rate (91.7%) among all groups (P<0.05). The GSH levels in the MF and Pre-SF(+)PACAP were significantly higher than in the other groups (P<0.05) and ROS levels was no significant difference among all groups. In conclusion, these results indicated that even though the oocytes were derived from SF, pre-IVM application of PACAP improved meiotic and cytoplasmic maturation by regulating intracellular oxidative stress.