• Title/Summary/Keyword: LKB

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Overexpression and Selective Anticancer Efficacy of ENO3 in STK11 Mutant Lung Cancers

  • Park, Choa;Lee, Yejin;Je, Soyeon;Chang, Shengzhi;Kim, Nayoung;Jeong, Euna;Yoon, Sukjoon
    • Molecules and Cells
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    • v.42 no.11
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    • pp.804-809
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    • 2019
  • Oncogenic gain-of-function mutations are clinical biomarkers for most targeted therapies, as well as represent direct targets for drug treatment. Although loss-of-function mutations involving the tumor suppressor gene, STK11 (LKB1) are important in lung cancer progression, STK11 is not the direct target for anticancer agents. We attempted to identify cancer transcriptome signatures associated with STK11 loss-of-function mutations. Several new sensitive and specific gene expression markers (ENO3, TTC39C, LGALS3, and MAML2) were identified using two orthogonal measures, i.e., fold change and odds ratio analyses of transcriptome data from cell lines and tissue samples. Among the markers identified, the ENO3 gene over-expression was found to be the direct consequence of STK11 loss-of-function. Furthermore, the knockdown of ENO3 expression exhibited selective anticancer effect in STK11 mutant cells compared with STK11 wild type (or recovered) cells. These findings suggest that ENO3-based targeted therapy might be promising for patients with lung cancer harboring STK11 mutations.

Effect of 24 h Fasting on Gene Expression of AMPK, Appetite Regulation Peptides and Lipometabolism Related Factors in the Hypothalamus of Broiler Chicks

  • Lei, Liu;Lixian, Zhu
    • Asian-Australasian Journal of Animal Sciences
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    • v.25 no.9
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    • pp.1300-1308
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    • 2012
  • The 5'-adenosine monophosphate-activated protein kinase (AMPK) is a key part of a kinase-signaling cascade that acts to maintain energy homeostasis. The objective of this experiment was to investigate the possible effects of fasting and refeeding on the gene expression of hypothalamic AMPK, some appetitive regulating peptides and lipid metabolism related enzymes. Seven-day-old male broiler (Arbor Acres) chicks were allocated into three equal treatments: fed ad libitum (control); fasted for 24 h; fasted for 24 h and then refed for 24 h. Compared with the control, the hypothalamic gene expression of $AMPK{\alpha}2$, $AMPK{\beta}1$, $AMPK{\beta}2$, $AMPK{\gamma}1$, Ste20-related adaptor protein ${\beta}$ ($STRAD{\beta}$), mouse protein $25{\alpha}$ ($MO25{\alpha}$) and agouti-related peptide (AgRP) were increased after fasting for 24 h. No significant difference among treatments was observed in mRNA levels of $AMPK{\alpha}1$, $AMPK{\gamma}2$, LKB1 and neuropeptide Y (NPY). However, the expression of $MO25{\beta}$, pro-opiomelanocortin (POMC), corticotropin-releasing hormone (CRH), ghrelin, fatty acid synthase (FAS), acetyl-CoA carboxylase ${\alpha}$ ($ACC{\alpha}$), carnitine palmitoyltransferase 1 (CPT-1) and sterol regulatory element binding protein-1 (SREBP-1) were significantly decreased. The present results indicated that 24 h fasting altered gene expression of AMPK subunits, appetite regulation peptides and lipometabolism related factors in chick's hypothalamus; the hypothalamic FAS signaling pathway might be involved in the AMPK regulated energy homeostasis and/or appetite regulation in poultry.

Ultrastructural Changes in the Neuropil of the Anterior Thalamic Nucleus following the Lesion in the Mamillary Body (유두체 손상이 시상전핵 조직상의 미세구조에 미치는 영향)

  • Lee, Byoung-Ho;Ko, Jeong-Sik;Ahn, E-Tay;Yang, Nam-Gil
    • Applied Microscopy
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    • v.18 no.2
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    • pp.177-186
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    • 1988
  • Degeneration of the axon terminals of mamillo-thalamic tract following the electrical coagulation of mamillary body is well known. In this study, the author investigated the ultrastructural alterations of neuropil components, initiated by terminal degenerations. Rats weighing approximately 250 gm were fixed on the stereotaxic instrument(David Kopf Inc., Heavy duty model), and NE 300 active electrode(Rhodes Med. Instr. Inc.) was introduced to the mamillary position of anterior 3.8 mm, lateral 0.5 mm, height 3.8 mm and lateral angle of $23^{\circ}$ according to De Groot's Atlas. Electric current of 20 mA was applied during 1 minute between active and inactive electrodes with Radio Frequency Lesion Generator(RFG 4, Radionics Inc.). Two hours, 2 days, 1 week and 2 weeks following the electrical coagulation of mamillary body, ipsilateral anterior thalamic nucleus was fixed in 1% glutaraldehyde-l% paraformaldehyde and 2% osmium tetroxide, embedded in Araldite mixture, cutted with LKB ultra tome V, stained with uranyl acetate-lead citrate and observed with JEOL 100 CX electron microscope. Observed results were as follows; 1. Degenerated mamillo-thalamic synapses were observed to form asymmetric axospinous or axo-dendritic types. 2. Terminal degeneration was not easily discernible at 2 hours interval after mamillary lesion, but following 2 days the terminal degeneration was apparent. 3. Postsynaptic spines, dendrites and even their cell bodies show edematic changes caused by the degeneration of postsynaptic counterpart. 4. Astrocytic territories, including perivascular processes forming glial limitans of blood-brain barrier, exhibit remarkable expansion. 5. Oligoglia and astroglia are actively engaged in the removal of degenerated elements. 6. Active forms of microglia were increased. 7. The observed results may represent typical ultrastructural alteration pattern within neuropil following the degeneration of certain input axon terminals.

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An Ultrastructural Study on the Duodenal Goblet Cells of Rabbit after Common Bile Duct Ligation (총담관 결찰이 집토끼 십이지장 점막내 배상세포의 미세구조에 미치는 영향)

  • Kim, In-Ho;Yang, Nam-Gil;Ahn, E-Tay;Ko, Jeong-Sik;Park, Kyung-Ho
    • Applied Microscopy
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    • v.20 no.1
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    • pp.51-64
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    • 1990
  • This experiment was performed to study the morphological changes of the goblet cells in the rabbit duodenal mucosa after common bile duct ligation. Healthy adult rabbits weighting about 2kg body weight were divided to normal and bile duct ligated groups. Common bile duct ligation was performed under ether anesthesia. Experimental animals were sacrificed on the 1st, 3rd, 5th, 7th and 14th day after the operation. Mucosal specimens from the upper part of duodenum were fixed in 2.5% glutaraldehyde-1.5% paraformaldehyde, followed by 1% osmium tetroxide, embedded in araldite mixture, cut with LKB-V ultratome, stained with uranyl acetate - lead citrate, and observed with JEM 100 CX II electron microscope. Observed results were as follow : 1. In the early stages(1st-5th day) of the experiments, the goblet cells showed apocrine and merocrine secretion. But those of the late stage(7th and 14th day) groups showed exocytotic merocrine secretion. 2. In the late stage of the experiments, there found than increase of newly formed goblet cells that contain electron lucent cytoplasms. 3. In the goblet cells of normal rabbit, mucous granules with higher or lower electron densities are found together in the cytoplasm, and electron lucent mucous granules occasionally fused together. But in the early stage of the common bile duct ligation, goblet cells contained granules of higher electron densities. 4. Considering the above findings, common bile duct ligation probably initiates the hypersecretion of mucous granules of goblet cells in the early stage, and may facilitate the differentiation of goblet cells in the later stage.

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A Study on the Fine Structure of Clonorchis sinensis, a Liver Fluke V. The Mature Spermatozoa (간흡충의 미세구조에 관한 연구 V. 정자의 미세구조)

  • 정계환;임한종
    • Parasites, Hosts and Diseases
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    • v.22 no.1
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    • pp.30-36
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    • 1984
  • An ultrastructural study on the mature spermatoBoa oi Cloptorchis sineitsis was carried out. For this study, the liver cukes were collected from the livers of rabbits and rats artificially infected with the metacercariae obtained from the fresh water fish, Pseudorasbora parve. Six-month old worms were used. The collected liver fiukes were washed with 0.85% saline solution and then immediately moved to cold 2% glutaraldehyde buffered with 0.1M Millonig's phosphate buffier (pH 7.4) . The materials were dissected into appropriate pieces in the fixative about 30 minutes after beginning of the fixation. Two hours later the materials containing the seminal receptacle were rinsed several times with the buffier and were secondarily fixed with cold, bugeyed 1% osmium tetroxide(OsO4) for 2 hours. The fully mixed tissue blocks Ivere dehydrated in a series of graded concentrations of acetone and were embedded in Epon 812 mixture. Thin sections obtained from LKB-5 ultramicrotome were stained with uranyl acetate and Reynold's lead citrate. Observations of the sections were carried out with JEM-100CX II electron microscope, In general, the mature sperm was long thread-like form with a sickle-shaped head. According to the longitudinal sectioned view of the sperm tail, the nucleus seemed to be spirally coiled and run a little far along the tail. The acrosome was not observed. The cytoplasm of the tail was biflagellated as usual in trematodes. Unlike other platyhelminth spermatozoa, the sperm tail of Clenorchis sinensis showed the ${\ulcorner}9+2{\lrcorner}$ in the microtubular arrangement. The mitochondria with poorly developed cristae were observed throughout the middle piece. The middle piece of the tail showed dull ladder or triangular shapes with the two flagella at the bottom. But, the principal piece of the tail was slightly flattened cylindrical shape with two aagella within the cytoplasm. The end piece was uniflagellated. It was not clearly identised whether the end piece was subdivided into two by aagellum or the lengths of the two aagella were different. The glycogen granules were rich in the cytoplasm throughout the lenght of the spermatozoa. These granules might be the energy source for the movement of the spermatosoa.

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Longitudinal Study on Taurine Intake of Breast-Fed Infants from Korean Non-Vegetarian and Lacto-Ovo-Vegetarian (모유 영양아의 수류기간별 Taurine 섭취량에 관한 연구)

  • 김을상;이종숙;최경순;조금호;설민영;박미아;이규한;이영남;노희경
    • Journal of Nutrition and Health
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    • v.26 no.8
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    • pp.967-973
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    • 1993
  • The longitudianl study on taurine intake of breast-fed infants from Korean lacto-ovo-vegetarian (n=23) and non-vegetarian(n=22) at 3-5, 15, 30, 60, 90, 120, 150 days postpartum was performed. The taurine content of human milk was analyzed by amino acid autoanalyzer (LKB-Alpha plus) and the human milk intak of infants was measured by test weighing method. The mean taurine content of human milk at each postpartum period was 434$\pm$128.362$\pm$, 304$\pm$90, 306$\pm$90, 302$\pm$77, 310$\pm$108, 248$\pm$53 nmol per ml in non-vegetarian and 418$\pm$112, 380$\pm$97, 314$\pm$97, 262$\pm$83, 206$\pm$79, 176$\pm$55, 153$\pm$31 nmol per ml in lacto-ovo-vegetarian, respectively. The content decreased significantly during lactation(p<0.05). The mean tauring intake at 15, 30, 60, 90, 120, 150 days postpartum was 195$\pm$94, 202$\pm$74, 212$\pm$54, 177$\pm$43, 137$\pm$47, 132$\pm$35, 106$\pm$15 umol per day in infants of lacto-ovo-vegetarian. Thu taurine content in human milk and the taurine intake of infants were different during lactation between non-vegetarian and lacto-ovo-vegetarian, and the intake per kg body weight during lactaion decreased.

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Studies on antibiotics resistance gene in Staphylococcus aureun Plasmid: Cloning of chloramphenicol resistance determinant (Staphylococcus aureus에서 분리된 plasmid상의 항생물질 저항성 인자에 관한 연구 : Chloramphenicol 저항성 인자의 클로닝)

  • 권동현;김영선;변우현
    • Korean Journal of Microbiology
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    • v.24 no.4
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    • pp.341-351
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    • 1986
  • R-plasmid(pSBK203, 2.5Mdal) conferring chloramphenicol resistance was isolated from mutiple antibiotic resistant Staphylococcus aureus D-H-1. Bacillus subtilis BD170 was transformed by this plasmid and restriction enzyme clevage sites of this plasmid were mapped for the cloning of chloramphenicol resistance gene. Taq I partial digested fragment of pSBK203(1.3kb) inserted into Cla I site of pBD9 appears to have both regulatory region for induction and structural gene for chloramphenicol resistance whereas Rsa I fragment (1.3kb, both ends are staggered away 0.1Kb from those of Taq I fragment) inserted into Sca I site of pBR322 showed constitutive expression in E. coli. Hinf I, Taq I, and Bgl II restriction enzyme recognition sites are found in both Rsa I fragment and Taq I fragment. Among these, Bgl II recognition site was associated with chloramphenicol resistance.

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Purification of Materials Produced by Amylocolatosis sp. and Anticancer Effect in Oral Cancer Model (Amylocolatosis sp.가 생산하는 항암물질의 정제 및 구강암 모델에 미치는 항암 효과)

  • Kim, Jung;Park, Young-Min;Lim, Do-Seon
    • Journal of dental hygiene science
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    • v.3 no.1
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    • pp.11-14
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    • 2003
  • A methylotrophic Actinomycetes strain, which produce the anti-oral cancer activity compound, was isolated from soil and estimated as Amylocolatosis sp. based on taxonomic studies. A methanol didn't have influence on the production of the anticancer compounds. These compound were isolated by ethylacetate extract, silica gel column chromatography, sephadex LH-20 column and reverse phase HPLC. The compounds were very stable under heat ($121^{\circ}C$), acid(pH 2.0) and alkali(pH 11.0) treatment. The cytotoxic effect of isolated anticancer compounds on various cancer cell lines such as A549, SNU-1, KB, L1210, and Sarcoma 180 was investigated by MTT assay method. And these produced compounds also showed the broad antimicrobial spectrum to test strains such as bacteria and yeast.

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Effect of Prunellae Spica on Oxidative Stress and Mitochondrial Dysfunction in the Hepatocyte (하고초(夏枯草)의 간세포에서 항산화 및 미토콘드리아 보호효과)

  • Jang, Mihee;Seo, Hye Lim;Kim, Sang Chan;Kim, Young Woo
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.30 no.1
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    • pp.20-26
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    • 2016
  • Prunellae Spica, the herbaceous plant in the genus Prunella, is a traditional herbal medicine and has been reported to have diuretic, anti-bacterial and anti-oxidant effects. However, the mechanism of its action was not clearly identified. In the present study, we investigated the hepatoprotective effect of Prunellae Spica extract (PSE) against the damage of mitochondria and death in hepatocyte induced by oxidative stress. Treatment of arachidonic acid (AA)+iron significantly induced oxidative stress and apoptosis in the hepatocytes. However, PSE protected cells and inhibited apoptosis by altering the protein levels such as poly(ADP-ribose) polymerase and pro-caspase 3. Moreover, AA+iron induced reactive oxygen species production and mitochondrial dysfunction, and Both of them were inhibited by PSE treatment. PSE markedly activated AMP-activated protein kinase (AMPK), an important regulator in cell survival. Furthermore, this activation by PSE was mediated with liver kinase B1, a major upstream kinase that phosphorylates Thr 172 of AMPKα, and this activation was associated with its cell protection, as assessed by an experiment of a chemical inhibitor. In conclusion, this study demonstrate that PSE protects hepatocytes against oxidative stress as mediated with activation of LKB1-dependent AMPK pathway.

Ultrastructural Influence of Nephrotoxic Agents on the Juxtaglomerular Cells of Mice (신장 상해물질이 생쥐 사구체곁세포의 미세구조에 미치는 영향)

  • Park, Kyung-Ho;Kim, Sang-Chul;Ahn, E-Tay;Ko, Jeong-Sik;Yang, Nam-Gil
    • Applied Microscopy
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    • v.26 no.4
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    • pp.431-446
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    • 1996
  • This experiment was performed to study the ultrastructural changes of the juxtaglomerular cell of mice following subcutaneous injection of heavy metallic agents. Male mice were divided into normal and experimental groups. The mice were subcutaneouly injected with $HgCl_2$ (2mg, 5mg or 10 mg/Kg/BW) or with $K_{2}Cr_{2}O_7$(5 mg, 10 mg or 20 mg/Kg/BW). Mice were sacrificed on 6 hours, 3 days and 14 days after the injection. Kidneys were fixed in the 2.5% glutaraldehyde-1.5% paraformaldehyde solution, followed by refixation in the 1% osmium tetroxide solution. Dehydrated blocks were embedded in araldite mixture. The sections were cut on a LKB-V ultratome, and ultrathin sections stained with uranyl acetate and lead citrate were observed with JEM 100CX II electron microscope. The results were as follow: 1. Juxtaglomerular cell of the experimental groups showed some alterations, especially in the structures of protein synthesis including dilations and degradations of granular endoplasmic reticula, atrophy of Golgi complex, and numerous free ribosomes in the cytoplasm. 2. Juxtaglomerular cells treated groups showed a number of vacuoles, protogranules and some myelin figures in the cytoplasm, especially in the earlier groups. 3. Juxtaglomerular cells of treated groups, contained a large number of secretory granules showing variable electron densities and pleomorphism in later groups (2 weeks). From the above results, it was concluded that, the mercuric chloride or potassium bichromate induces acute renin release from juxtaglomerular cells of the mice, but many juxtaglomerular cells may secrete prematured secretory granules, or the synthetic system of the cell can not perform normal function.

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