• 제목/요약/키워드: membrane aggregation

검색결과 79건 처리시간 0.028초

유기인계 및 카바메이트계 농약 신속 검출을 위한 역 Y자 스트립의 개발 (Development of an Inverted Y-Shaped Strip for the Detection of Organophosphorus and Carbamate Residual Pesticides)

  • 이정은;김솔아;박현진;문효영;심원보
    • 한국식품위생안전성학회지
    • /
    • 제36권1호
    • /
    • pp.1-8
    • /
    • 2021
  • 본 연구에서는 농약의 음성시료에서는 acetylcholinesterase와 acetylthiocholine을 반응시켜 +전하와 -전하를 가지는 thiocholine으로 분해되어 금 나노입자를 응집시켜 역 Y자 스트립상에서 청자색의 반응선(띠)을 형성하고 양성 시료에서는 생성시키지 않는 원리를 이용한 신속 농약 검출법을 개발하였다. 개발한 분석법은 유기인계 농약 말라옥손과 카바메이트계 농약 카보퓨란을 각각 10 ng/mL 수준까지 검출이 가능한 것으로 확인되었으며, 2종의 유기인계와 카바메이트계 농약(EPN, dichlorvos)에 대해 추가적으로 검출 한계를 확인한 결과에서도 10 ng/mL 수준까지 모두 검출 가능함을 확인할 수 있었다. 그러나 3종의 트리아진 계열의 농약과 각 1종의 피레스로이드, 카복사마이드, 페닐아마이드 및 유기염소계열의 농약에 대해서는 반응성이 없는 것으로 확인되어 유기인계와 카바메이트계 농약 분석에 적용이 가능한 것으로 확인되었다. 마지막으로, 임의로 오염시킨 농산물 시료를 대상으로 분석법의 회수율을 확인한 결과, 말라옥손에 대해서 96.4에서100.7%, 카보퓨란은 81에서112.7%의 회수율이 확인되어 본 연구에서 개발한 역 Y자 스트립을 농약 검출법으로 이용한다면 농산물과 농업환경 중 존재하는 유기인계 및 카바메이트계 잔류농약을 신속하게 검출할 수 있을 것으로 판단된다.

Alcohol Fermentation at High Temperature and the Strain-specific Characteristics Required to Endow the Thermotolerance of Sacchromyces cerevisiae KNU5377

  • Paik, Sang-Kyoo;Park, In-Su;Kim, Il-Sup;Kang, Kyung-Hee;Yu, Choon-Bal;Rhee, In-Koo;Jin, In-Gnyol
    • 한국미생물생명공학회:학술대회논문집
    • /
    • 한국미생물생명공학회 2005년도 2005 Annual Meeting & International Symposium
    • /
    • pp.154-164
    • /
    • 2005
  • Saccharomyces cerevisiae KNU5377 is a thermotolerant strain, which can ferment ethanol from wasted papers and starch at 40$^{\circ}C$ with the almost same rate as at 30$^{\circ}C$. This strain showed alcohol fermentation ability to convert wasted papers 200 g (w/v) to ethanol 8.4% (v/v) at 40$^{\circ}C$, meaning that 8.4% ethanol is acceptable enough to ferment in the industrial economy. As well, all kinds of starch that are using in the industry were converted into ethanol at 40$^{\circ}C$ with the almost same rate as at 30$^{\circ}C$. Hyperthermic cell killing kinetics and differential scanning calorimetry (DSC) revealed that exponentially growing cells of this yeast strain KNU5377 were more thermotolerant than those of S. cerevisiae ATCC24858 used as a control. This intrinsic thermotolernace did not result from the stability of entire cellular components but possibly from that of a particular target. Heat shock induced similar results in whole cell DSC profiles of both strains and the accumulation of trehalose in the cells of both strains, but the trehalose contents in the strain KNU5377 were 2.6 fold higher than that in the control strain. On the contrary to the trehalose level, the neutral trehalase activity in the KNU5377 cells was not changed after the heat shock. This result made a conclusion that though the trehalose may stabilize cellular components, the surplus of trehalose in KNU5377 strain was not essential for stabilization of whole cellular components. A constitutively thermotolerant yeast, S. cerevisiae KNU5377, was compared with a relatively thermosensitive control, S. cerevisiae ATCC24858, by assaying the fluidity and proton ATPase on the plasma membrane. Anisotropic values (r) of both strains were slightly increased by elevating the incubation temperatures from 25$^{\circ}C$ to 37$^{\circ}C$ when they were aerobically cultured for 12 hours in the YPD media, implying the membrane fluidity was decreased. While the temperature was elevated up to 40$^{\circ}C$, the fluidity was not changed in the KNU5377 cell, but rather increased in the control. This result implies that the plasma membrane of the KNU5377 cell can be characterized into the more stabilized state than control. Besides, heat shock decreased the fluidity in the control strain, but not in the KNU5377 strain. This means also there's a stabilization of the plasma membrane in the KNU5377 cell. Furthermore, the proton ATPase assay indicated the KNU5377 cell kept a relatively more stabilized glucose metabolism at high temperature than the control cell. Therefore, the results were concluded that the stabilization of plasma membrane and growth at high temperature for the KNU5377 cell. Genome wide transcription analysis showed that the heat shock responses were very complex and combinatory in the KNU5377 cell. Induced by the heat shock, a number of genes were related with the ubiquitin mediated proteolysis, metallothionein (prevent ROS production from copper), hsp27 (88-fold induced remarkably, preventing the protein aggregation and denaturation), oxidative stress response (to remove the hydrogen peroxide), and etc.

  • PDF

Laminin-1 Phosphorylation by Protein Kinase A: Effect on self assembly and heparin binding

  • Koliakos, George;Kouzi-Koliakos, Kokkona;Triantos, Athanasios;Trachana, Varvara;Kavoukopoulos, Evaggelos;Gaitatzi, Mary;Dimitriadou, Aphrodite
    • BMB Reports
    • /
    • 제33권5호
    • /
    • pp.370-378
    • /
    • 2000
  • Incubation of purified laminin1-nidogen1 complexes with $[{\gamma}-^{32}P]-ATP$ in the presence of the catalytic subunit of the protein kinase A (cAMP-dependent protein kinase) resulted in the phosphorylation of the alpha chain of laminin-1 and of the nidogen-1 molecule. Aminoacid electrophoresis indicated that phosphate was incorporated on serine residues. The phosphorylation effect of laminin-1 on the process of self assembly was studied by turbidometry. In these experiments, the phosphorylated laminin-1 showed a reduced maximal aggregation capacity in comparison to the non-phosphorylated molecule. Examination of the laminin-1 network under the electron microscope showed that the phosphorylated sample formed mainly linear extended oligomers, in contrast to controls that formed large and dense multimeric aggregates. Heparin binding on phosphorylated laminin-1 in comparison to controls was also tested using solid-phase binding assays. The results indicated an enhanced heparin binding to the phosphorylated protein. The results of this study indicate that laminin1-nidogen1 is a substrate for protein kinase A in vitro. This phosphorylation had an obvious influence on the lamininl-nidogen1 network formation and the heparin binding capacity of this molecule. However, further studies are needed to investigate whether or not this phenomenon could play a role in the formation of the structure of basement membranes in vivo.

  • PDF

수종(數種) 항균제(抗菌劑) 처리(處理)에 의(依)한 Escherichia coli의 미세구조(微細構造) 변화(變化) (Cytological Changes Associated with the Exposure of Escherichia coli to Several Disinfectants: An Ultrastructural Study)

  • 등영건;고춘명;김성광
    • 대한미생물학회지
    • /
    • 제11권1호
    • /
    • pp.1-11
    • /
    • 1976
  • Escherichia coli(ATCC 11115)에 실험실등에서 상용하는 여러가지 항균제를 시간별로 처리, 그 변화양상을 전자현미경으로 관찰한 바 그 결과를 요약하면 다음과 같다. 1) 대조군은 3층의 단일막으로 형성된 세포벽에 둘러쌓여 있으며 세포질은 전자밀도가 낮은 nucleoid와 ribosme들이 산재하여 있음을 관찰할 수 있었다. 2) 70% ethanol용액 처리군은 핵물질을 관찰할 수 없었고 세포질은 세포 중앙부로 응집되어 있었으며 세포벽의 외부에서는 bleb 들을 관찰 할 수 있었다. 3) 3% $H_2O_2$ 용액 처리군은 세포내용물의 변화는 70% ethanol 처리군과 대동소이(大同小異)하였으나 세포벽에서는 심한 굴곡현상이 관찰되었다. 4) 5% lysol 용액처리군은 세포질 및 핵물질 부위가 완전히 구분되어 나타났으며 세포질내의 ribosome과립들은 시간이 경과할수록 그 응집현상이 심하였고 세포 외부에는 ribosome 양 과립들이 부착하고 있음이 관찰되었다. 5) 1% DDEGH 용액 처리군은 세포질의 응집 및 세포막과 세포벽이 뚜렷이 관찰되지 않았으며 세포외부에 세포내용물과 동일한 물질로 생각되는 물질이 부착되어 있음을 관찰할 수 있었다. 6) 고압멸균 처리군은 세포막 및 세포벽의 파괴, 탈락 및 세포내용물의 유출현상이 관찰되었다.

  • PDF

폐렴구균 알코올탈수소효소의 세포 특이성 및 세포내 분포 (Immunological Characterization and Localization of the Alcohol-dehydrogenase in Streptococcus pneumoniae)

  • 권혁영;박연진;표석능;이동권
    • 미생물학회지
    • /
    • 제37권3호
    • /
    • pp.221-227
    • /
    • 2001
  • 열충격 단백질(heat shock protein: HSP)은 변성된 단백질의 응집을 방지하여 가혹한환경에서 병원균의 생존을 증가시킨다. 세균에 알코을 stress를 가하면 다량의 DnaK와 GronEL이 유도되지만 폐렴구균에서는 DnaK와 GroEL이 전혀 유도되지 않는 대신 알코올탈수소효소(alcohol dehydrogenase : ADH)가 유도되었다. 이런 특성은 폐렴구균 ADH가 HSP처럼 chaperone 기능을 수행라고 있을 가능성을 제시하고 있으므로 본 연구에서는 일차적으로 ADH 유전자를 확인하고 ADH 의 면역특성 및 세포내 분포를 측정하였다. 폐렴구균 ADH는 이질아메바 ADH2 및 대장균 ADH 와 높은 유사성을 나타냈으며 883 개의 아미노산으로 구성된 등전점 6.09의 단백질로 추정된다. 그러나 폐렴구균 ADH와 유사성이 높은 대장균, 유산균 및 황색포도상구균의 용해액을 폐렴구균 ADH 항체와 immunoblot을 실시하였을 때 전혀 반응하지 않았다. 또한 세포질, membrane, periplasm에 있는 단백질 분획 및 폐렴구균 배양 상등액을 ADH 항체와 immune blot을 실시하였을 때 ADH 는 열충격에 관계없이 세포 밖으로 분비되는 단백질임을 확인하였다. 이런 결과는 폐렴구균 ADH가 진단용항원 및 백신으로 개발될 수 있는 가능성을 제시하고 있다.

  • PDF

성견에서 조직유도재생술과 구연산, 테트라싸이크린으로 치근면 처치한 경우 치주조직의 치유에 미치는 영향 (THE EFFECTS OF ROOT TREATMENT WITH CITRIC ACID AND TETRACYCLINE AND GUIDED TISSUE REGENERATION ON THE PERIODONTAL HEALING RESPONSES IN DOGS)

  • 김주환;김종여;정진형
    • Journal of Periodontal and Implant Science
    • /
    • 제24권1호
    • /
    • pp.87-97
    • /
    • 1994
  • The ultimate goal of periodontal therapy is to fully reconstruct the periodontal attachment apparatus. Commonly used techniques for treatment of infrabony defects include a combination of root planing, curettage and root treatment. To prevent the apical migration of epithelial cells, the technique of guided tissue regeneration is used. The aim of this study is to compare the effects of root treatment with Citric acid & Tetracycline and Guided tissue regeneration in dogs. Experimental periodontitis was induced by the ligation of orthodontic elastic threads in the upper right and left premolars 3, 4 of five adult dogs for 6 weeks. 4 types of procedures were performed as follows; 1) Control graup : Mucoperiosteal flap 2) Experinental I : GTR used Gore-tex(R) membrane 3) Experinental II : Root treatment with citric acid (PHl) 4) Experinental III : Root treatment with tetracycline HCl (50mg/ml) There after, dogs were serially sacrificed at the 1, 2, 4, 5, 8 weeks, and the specimens were prepared, and stained with hematoxylin-eosin for the light microscopic evaluation. The results of this study were as follows; 1. Junctional epithelium reached to the notch through the furcation area in control group at 8 weeks. 2. In the aspects of the inflammatory cell infiltration, control group showed severe aggregation than experimental group I, II, III through the experimental period 3. New cementum was observed over the notch from 5 weeks in experimental group II 4. In the aspects of the amount of new bone formation, experimental group was better than control group, but there was not significant differences among the experimental group, I, II, III

  • PDF

Inhibition of Melanosome Transport by Inducing Exon Skipping in Melanophilin

  • Jin Young Kim;Seon-Young Han;Kiho Sung;Jeong Yeon Seo;Cheol Hwan Myung;Chan Song Jo;Jee Hoe Yoon;Ji Yun Park;Jae Sung Hwang
    • Biomolecules & Therapeutics
    • /
    • 제31권4호
    • /
    • pp.466-472
    • /
    • 2023
  • Exon skipping is an efficient technique to inhibit specific gene expression induced by a short-sequence peptide nucleic acid (PNA). To date, there has been no study on the effects of PNA on skin pigmentation. In melanocytes, the tripartite complex is responsible for the transport of mature melanosomes from the nucleus to the dendrites. The tripartite complex is composed of Rab27a, Mlph (Melanophilin), and Myosin Va. Defects in the protein Mlph, a melanosome transport-related protein, are known to cause hypopigmentation. Our study shows that Olipass peptide nucleic acid (OPNA), a cell membrane-permeable PNA, targets exon skipping in the Mlph SHD domain, which is involved in Rab27a binding. Our findings demonstrate that OPNA induced exon skipping in melan-a cells, resulting in shortened Mlph mRNA, reduced Mlph protein levels, and melanosome aggregation, as observed by microscopy. Therefore, OPNA inhibits the expression of Mlph by inducing exon skipping within the gene. These results suggest that OPNA, which targets Mlph, may be a potential new whitening agent to inhibit melanosome movement.

파동형 Nd:YAG 레이저조사가 인체 치은조직의 미세구조에 미치는 영향 (The effect of a pulsed-Nd:YAG laser irradiation on microstructure of human gingiva)

  • 한경윤;신광용;김천석;김형수;염창엽;김병옥
    • Journal of Periodontal and Implant Science
    • /
    • 제27권2호
    • /
    • pp.317-328
    • /
    • 1997
  • Since laser therapy has been applied to dentistry, many dental practitioners are very interested in laser therapy on various intraoral soft tissue lesions including gingival hyperplasia and aphthous ulcer. The purpose of the present study was to determine the therapeutic effect and the harmful effect of a pulsed-Nd:YAG laser irradiation on human gingival tissue. In twenty periodontal patients with gingival enlargement, the facial gingival surface of maxillary anterior teeth was randomly irradiated at various power of 1.0W(100mJ, 10Hz), 3.0W(100mJ, 30Hz) and 6.0W(l50mJ, 40Hz) for 60 seconds by contact delivery of a pulsed-Nd:YAG laser(EN.EL.EN060, Italy). Immediately after laser irradiation, the gingival tissues were surgically excised and prepared in size of 1mm3. Subsequently the specimens were processed for prefixation and postfixation, embedded with epon mixture, sectioned in $1{\mu}$ thickness, stained with uranyl acetate and lead citrate, and observed under transmission electron microscope(JEM 100 CXII). Following findings were observed; l. In the gingival specimens irradiated with l.OW power, widening of intercelluar space and minute vesicle formation along the widened intercellular space were noted at the epithelial cells adjacent to irradiated area. 2. In the gingival specimens irradiated with 3.0W power, the disruption of cellular membrane, aggregation of cytoplasm, and loss of intercellular space were observed at the epithelial cells adjacent to irradiated area. 3. In the gingival specimens irradiated with 6.0W power, the disruption of nuclear and cellular membrane was observed at the epithelial cells adjacent to irradiated area. The ultrastructural findings of this study suggest that surgical application of a pulsed-Nd:YAG laser on human gingival tissue may lead somewhat delayed wound healing due to damage of epithelial cells adjacent to irradiated area.

  • PDF

생쥐 배아 동결시 액체질소의 분사속도가 해빙후 배아의 발달, 미세섬유, 미토콘드리아 및 세포자연사에 미치는 영향 (Effect of Different Infusion Frequency of Liquid Nitrogen on Actin Filament, Mitochondria, Apoptosis and Development in Mouse 2-Cell Embryo after Freezing and Thawing)

  • 손인표;안학준;계명찬;최규완;민철기;강희규;이호준;권혁찬
    • 한국발생생물학회지:발생과생식
    • /
    • 제4권2호
    • /
    • pp.161-173
    • /
    • 2000
  • 컴퓨터 세포동결기를 이용하여 생쥐 배아를 동결할 때 액체질소 (L$N_2$)의 분사속도가 해빙 후 배아의 미세구조, 기능 및 발달에 미치는 영향을 알아보고자 하였다. 이를 위해 배아는 동결을 하지 않은 대조군 (control) 및 동결군에서 L$N_2$의 분사속도에 따라 고속분사군 (120 infusion/min group 1), 저속분사군 (50 infusion/min; group 2)으로 나누었다. ICR 계열의 생쥐의 2 세포기 배아를 사용하였으며, 동결 및 해빙은 저속동결-급속해빙 방법을 사용하였다. 각 군에 따라 해빙 후 배아의 생존율과 세포질이 양호하고 분절화가 없는 2세포기 배아를 대상으로 포배 발달율 및 할구수를 측정하였다. 공초점 현미경을 이용하여 배아 내에서의 $H_2O$$_2$, 활성 미토콘드리아의 분포, 막전위차 및 actin filament를 측정하였으며, TUNEL 방법을 이용하여 DNA 분절화를 확인하였다. 동결-해빙 후 건강한 2 세포기 배아의 회수율은 group 1 (50.7%)에 비해 group 2 (34.6%)에서 현저히 감소했다 (p<0.05). 포배기 배아의 발생율 (86.7%, 76.7% vs. 44.0%)과 할구수 (79.5$\pm$12.9, 71.6$\pm$8.0 vs. 62.5$\pm$4.7)는 대조군 혹은 group 1에 비해 group 2에서 유의한 차이를 보였다 (p<0.05). H$_2$0$_2$의 상대적 강도는 group 2에서 유의하게 증가하였다 (15.3$\pm$3.0, 16.6$\pm$1.6 vs. 23.4$\pm$1.8, p<0.05). 활성 미토콘드리아의 분포는 정상적인 배아에서는 균등하게 분포하는 반면 배발달이 정지된 배아에서는 원형질막 주위에 몰리고 응집된 양상을 보였다. 그러나 대조군, group 1, group 2에서는 모두 균등하게 분포하여 각 군간에 차이가 없었다. 미토콘드리아의 JC-1 염색 결과는 대조군과 group 1의 경우 590 nm의 파장으로 발산되는 미토콘드리아가 group 2에 비하여 유의하게 증가하였다 (17.2$\pm$3.8, 17.4$\pm$1.3 vs. 13.2$\pm$2.0, p<0.05). 2세포기 배아내 미세섬유 (actin filament)는 대조군 및 group 1의 경우 균일하게 분포하는 반면, group 2에서는 부분적인 결손과 응집현상이 관찰되었다. DNA 분절율 (30.8%, 36.0% vs. 65.6%; p<0.05)은 group 2에서 유의하게 증가하였다. 동결시 액체질소의 분사속도는 해빙 후 배아 발달에 매우 중요한 요인으로 작용하며, L$N_2$의 분사속도의 증가는 동결과 정에서 하강 온도의 미세한 변화를 감소시켜 세포내 골격구조와 미토콘드리아의 상해를 감소시켜 $H_2O$$_2$의 발생과 DNA 분절화를 감소시켜 배아 발생을 호전시키는 것으로 사료된다.

  • PDF

Telluric Acid가 흰쥐 간조직의 미세구조에 미치는 영향 (Ultrastructural Changes Induced by Telluric Acid in the Rat Liver)

  • 손석주;정영길;조승묵;백태경;최창도;최월봉
    • Applied Microscopy
    • /
    • 제25권4호
    • /
    • pp.83-103
    • /
    • 1995
  • This experiment was carried out to investigate the effects of telluric acid on the histological and fine structural changes in the rat liver. Fischer 344 rats($150{\sim}200gm$) were used in this study as control and experimental groups. Telluric acid(5 mg/100 gm of body weight) suspensed in olive oil was given intraperitoneally to the animals of the experimental group and only olive oil to those of the control group. At the intervals of 3, 6 and 12 hours, 1, 2, 3, 5, 10, 20, 30 and 60 days after administration, the animals were sacrificed, and livers were obtained from the rats. For light microscopic examination of the liver, sections($5{\mu}m$) were stained with hematoxylineosin(H-E). For electron microscopic examination of the liver, sections were stained with uranyl acetate and lead citrate, finally examined with Zeiss EM 109 electron microscopes. The results obtained were as follows. 1. In the control group, round nucleus. well developed mitochondria, Golgi apparatus, rough endoplasmic reticulum(RER) and numerous glycogen particles were observed in the cytoplasm of the hepatocyte. In the cytoplasmic membranes of the hepatocyte, sinusoidal surface had numerous microvilli and cellular surface is combinated adjacent hepatocyte with desmosomes. The RER cisterns were dilated and zymogen granules were fewer than those of the dark cells. Kupffer cells with irregular nuclear membrane were observed. Fat storing cell and collagenous fiber bundle were observed in the Disse space. 2. Kupffer cell, inflammatory cells in the connective tissue of hepatic triad and lysosome were increased in the 3, 6, and 12 hour experimental group comparing with that of the control group. 3. In the 1 day experimental group, infiltration of inflammatory cells in interlobular connective tissue, dilatation of sinusoidal capillary and increasing of Kupffer cell were observed. Atropic change of hepatocyte and aggregation of glycogen particles in the cytoplasm of hepatocyte were observed. In this group, desmosome near bile canaliculi and collagenous fiber bundle in the Disse space were increased comparing with that of the 12 hours experimental group. In the 2 days experimental group, desmosome, lysosome, peroxisome and collagenous fiber bundle were increased comparing with that of the 1 day experimental group. Furthermore, lamellated bodies were also seen in the cytoplasm of the hepatocyte. 4. In 3 and 5 days experimental groups, transformations of hepatic cell cord and degeneration of the hepatocyte were markedly inclosed comparing with the all experimental groups. And damaged RER and mitochondria. collagenous fiber bundle were also inclosed comparing with that of the 2 days experimental group. Autophagosome and fat storing cells with large lipid droplets were also observed comparing with that of the 2 days experimental group. Tight junction and desmosome between the hepatocytes were separated. These degenerating changes were severe through the all experimental groups. 5. In the 10 and 20 days experimental groups, arrangement of hepatic cell cords and cell organelles of hepatocytes were similar to those of the control group. However, aggregation of glycogen particles, dilatation of sinusoidal capillary and infiltration of inflammatory cells remained. 6. In the 30 days experimental group, the tissue findings were similar to those of the control grout. But lamellated bodies in some hepatocytes and lysosome were remained in the cytoplasms of the Kupffer cells. In the 60 days experimental group, these all changes were recovered as the control group. In conclusion, telluric acid would directly induce the degenerative and necrotic changes on the hepatic tissue. However, these changes were perfectly recoverd in the 60 days experimental group as the control group.

  • PDF