• Molecules and Cells
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• 제45권3호
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• pp.134-147
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• 2022

The anti-oxidant enzyme heme oxygenase-1 (HO-1) is known to exert anti-inflammatory effects. From a library of pyrazolo[3,4-d]pyrimidines, we identified a novel compound KKC080096 that upregulated HO-1 at the mRNA and protein levels in microglial BV-2 cells. KKC080096 exhibited anti-inflammatory effects via suppressing nitric oxide, interleukin1β (IL-1β), and iNOS production in lipopolysaccharide (LPS)-challenged cells. It inhibited the phosphorylation of IKK and MAP kinases (p38, JNK, ERK), which trigger inflammatory signaling, and whose activities are inhibited by HO-1. Further, KKC080096 upregulated anti-inflammatory marker (Arg1, YM1, CD206, IL-10, transforming growth factor-β [TGF-β]) expression. In 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridinetreated mice, KKC080096 lowered microglial activation, protected the nigral dopaminergic neurons, and nigral damage-associated motor deficits. Next, we elucidated the mechanisms by which KKC080096 upregulated HO-1. KKC080096 induced the phosphorylation of AMPK and its known upstream kinases LKB1 and CaMKKbeta, and pharmacological inhibition of AMPK activity reduced the effects of KKC080096 on HO-1 expression and LPS-induced NO generation, suggesting that KKC080096-induced HO-1 upregulation involves LKB1/AMPK and CaMKKbeta/AMPK pathway activation. Further, KKC080096 caused an increase in cellular Nrf2 level, bound to Keap1 (Nrf2 inhibitor protein) with high affinity, and blocked Keap1-Nrf2 interaction. This Nrf2 activation resulted in concurrent induction of HO-1 and other Nrf2-targeted antioxidant enzymes in BV-2 and in dopaminergic CATH.a cells. These results indicate that KKC080096 is a potential therapeutic for oxidative stress-and inflammation-related neurodegenerative disorders such as Parkinson's disease.

• 한국동물학회지
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• 제26권4호
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• pp.271-282
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• 1983

동면기 양서류 피부 색소세포의 미세구조를 관찰하기 위하여 무미 양서류인 개구리 (Rana nigromaculata)의 피부조직을 2.5% glutaradehyde-paraformaldehye (pH 7.2)와 2% osmium tetroxide에 전후 고정한 후 ethanol과 acetone으로 탈수, Epon 812 mixture에 포매하여 LKB ultramicrotome으로 초박절 표본을 만들어 uranyl acetate와 lead citrate로 염색하여 Jeol-100B형 전자현미경으로 관찰하여 다음과 같은 결과를 얻었다. 동면기 개구리의 피부 색소세포는 대황세포, iridophore 및 흑색소 보유세포로 구성되었으며, 이들 세포의 특징은 다음과 같다. 1. 대황세포 A. 대황세포는 pterinosome과 carotinoid vesicles이 전세포질에 채워져 있었으며, 많은 ribosome과 소수의 mitochondria 및 glycogen particle이 pterinosome 사이에 분산되었다. B. Pterinosome은 크고 작은 원형 또는 타원형이며, 이 소낭속의 내부 구조물에 EK라 6가지 형 (제1형 pterinosome, 제2형 pterinosome, 제3형 pterinosome, 제4형 pterinosome, 제5형 pterinosome, 제6형 pterinosome)으로 구분되며, 특히 제1형 pterinosome, 제2형 pterinosome, 제3형 pterinosome이 발달되었따. C. Carotinoid vesicle은 작은 원형 또는 타원형이며, 대부분의 carotinoid vesicle은 핵 주변부에 덩어리 모양으로 모여있고, 일부분은 이 세포의 pterinosome 사이에 분산되어 나타났다. 2. Iridophore A. Iridophore는 볼록렌즈와 같이 나타나고 좁은 세포간 공간에 의하여, 대황세포 아래에 위치하였다. B. Iridophore는 장방형 EH는 방추형의 reflective platelet로 채워져 있었으며, 서로 평행하게 규칙적으로 배열되었다. 3. 흑색소 보유세포 A. 흑색소 보유세포는 대황세포와 iridophore와 평행하여 가장 밑부분에 위치하였다. B. 흑색소 과립은 원형 또는 타원형으로 전세포질에 채워져 있었으며, 세포 소기관은 잘 관찰되지 않았다. C. 흑색소 과립으로 채워진 흑색소 보유세포의 돌기는 iridophore의 양 측부로 \ulcorner어 올라가 대황세포와 iridophore의 사이에 위치하였다.

• 한국동물학회지
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• 제22권3호
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• pp.103-114
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• 1979

양서류 피부과립선의 미세구조를 관찰하기 위하여 무미양서류인 무당개구리(Bombina orientalis), 두꺼비 (Bufo bufo gargarizans), 개구리 (Rana nigromaculata) 및 옴개구리 (Rana rugosa)의 피부조직은 2.5% glutaraldehyde-paraformaldehyde (pH 7.2)와 1%osmium tetroxide에 전후 고정한 후 ethanol과 acetone으로 탈수, Epon 812 mixture에 포매하여 LKB ultramicrotome으로 초박절표본을 만들어 uranyl acetate와 lead citrate으로 염색하여 JEOL-100B형 전자현미경으로 관찰하여 다음과 같은 결과를 얻었다. 1. 양서류 피부과립선은 선상피세포와 근상피세포로 이루어졌다. 2. 양서류 피부과립선의 선상피세포는 암세포들로 구성되었으나 무당개구리에서는 명세포도 관찰되었다. 3. 양서류 피부과립선은 전분지를 하였다. 4. 양서류 피부과립선 상피세포의 핵은 원형 내지 타원형으로 크고 작은 핵경함요를 보였고, heterochromatin은 주로 핵경 인접부에 많았다. 세포질에는 mitochondria가 핵주변부에 비교적 많이 산재하였고, rough-surfaced endoplasmic reticulum은 핵 주변부에 발달하였지만 smooth-surfaced endoplasmic reticulum은 미약하였다. 5. 분비과립들은 구형 또는 난형으로 높은 전자밀도를 보였으며 다소 약한 전자밀도를 보이는 과립도 관찰되었다. 6. 양서류 과립선내 분비과립들이 같은 세포내에서 다소 전자밀도의 차이를 보이는 것은 분비과립의 형성단계에 따른 농도 차이에 기인하며 그 화학적 조성은 유사하다고 생각된다.

• Journal of Oral Medicine and Pain
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• 제8권1호
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• pp.33-41
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• 1983

The authors observed the ultrastructure of oral leukoplakia simplex of gingiva, buccal mucosa, tongue and alveolar ridge. For the purpose of clearly defining the lesions under investigation in this study, leukoplakias were cinsidered to be any white patches on the oral mucous membranes that could not be removed by rubbing and could not be classified clinically or microscopically as another diagnosable disease. The tissue to be examined were embedded in paraffin for light microscopic study. The tissue to be examined under the electronomicroscope were fixed in 2.5% glutaraldehyde in 0.1M cacodylate buffer and 1% osmic acid in 0.1M cacodylate buffer, dehydrated with guaded alchol, and treated with propylene oxide, and embedded in Epon.Ultrathin sections were obtained by LKB III ultrotome, stained with uranyl acetate/lead citrate, and examined with Corinth 500EM. The results were as follows : 1. Epithelium of leukoplakia consisted of stratum basale, stratum spinosum, stratum granulosum and stratum corneum. 2. There was hyperorthokerotosis or hyperparakeratosis. 3. Granular cells contained a lot number of membrane coating granule showing lamellar structure, clearing ot codensation, and a lot of keratohyaline granule varied in size. 4. An increased concentration of tonofilaments and an increased number of desmosomes were found in the stratum spinosum. 5. Basal lamina generally showed its continuity, but in some locatoins, its interreption and multiplication appeared.

• 한국식품영양과학회지
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• 제28권1호
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• pp.9-15
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• 1999

Taurine contents in alcoholic and non alcoholic beverages, milk and milk products, sugars and condiments were determined for 83 food items commonly used by Koreans. Taurine concentrations of food samples were analyzed by using an automated amino acid analyzer(Biochrom 20, Pharmacia LKB) based on ion exchange chromatography. Taurine was not detected in most carbonated beverages, but found in a variety of fruit and vegetable beverages(canned) in the range of 0.13 ~1.43mg taurine/ 100g wt, and in instant coffee or tea beverages(canned) in the range of 0.10~0.41mg taurine/100g wt. The traditional Korean turbid rice wine(takju) sample contained the highest level of taurine (2.29mg/100g wt) among alcoholic beverages tested, which was followed by wine(0.59~0.88mg tau rine/100g wt) and beer(0.53~0.73mg taurine/100g wt). Ordinary milk samples contained 1.05~1.40 mg taurine/100g wt, and a variety of taurine supplemented infant formulas manufactured by Korean companies included 17.3~25.4mg taurine/100g wt. Taurine was not detected in most sugars and sweetners except a couple of chocolate samples(2.13~2.18mg taurine/100g wt). Exceptionally high level of taurine was found in a curry powder sample(16.9mg taurine/100g wt), while the rest of commonly used condiments contained less than 2.90mg taurine/100g wt.

• 대한치과의사협회지
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• 제10권5호
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• pp.307-316
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• 1972

신생 Ball/c Strain 종 웅성백서 28두를 사용하고, 28두중 14두는 실험군에, 나머지 14두는 대조군에 각각 배당하였다. 실험군과 대조군은 동물희생시간(30분, 1시간, 4시간, 8시간, 12시간, 24시간 48시간)에 따라서 7군으로 구분하고, 각군에 2두식 배정하였다. 실험군은 백서를 Bell Jar내에 넣고, 여기에 창소를 서서히 넣어 Westinghouse Oxygen Analyzer로 30내지 40ppm이 되도록 하고 15분 동안 방치한 후, 이를 밖으로 제거하여 신선한 공기를 5분동안 마시게 하였다. 이런 과정을 반복하여 결국 55분 동안에 45분간 무 산소 상태에 있게 하였다. 악하선의 소편을 2% Paraformaldehyde(0.1M Cacodylate fuffer) 용액에 4시간동안 고정하였고, 탈수후 전현 표본포리법에 따라 Epoxy유지를 혼합하여 이몰하였다. LKB Ultratome으로 1μ의 연속절편을 만들었고 야기시킨후 신생백서의 악하선세포에 세포학적변화를 보고저하였고, 기결과를 요약하면 다음과 같다. 1. 신생 백서에 산소결핍을 야기시켰을때에 나타나는 초기의 세포학적변화는 이를 이르킨 후 1시간과 4시간사이가 절정에 달하였다. 이실험에서 유해적 영향은 공포화하고 점액성 및 장액성세포의 핵이 농축변성하며 장액성분필과기의 방출이 지연되어 세포질첨단에 축적되었다. 이들의 회복은 적어도 4시후에 시작되과, 12시간에는 세포질의 대부분, mrgl 핵의 용태가 정상으로 되돌아간다. 24시간에는 실험군과 대조군간에 있어서 야간의 차이는 인정되나 선소엽은 아직 초기에 볼수있었든 퇴행성변화의 잔여물을 함유하고 있었다. 2. 위에 요약한 본연구의 결과는 노인들이 보고한 산소결핍증이 악하선에 있어서도 단백질합성의 억제현상을 나타냈음을 세포학적으로 확증한 것이라하겠다.

• Applied Microscopy
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• 제18권2호
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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.

• Parasites, Hosts and Diseases
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• 제22권1호
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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.

• 치위생과학회지
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• 제3권1호
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• pp.11-14
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• 2003

각종 토양시료로부터 M배지와 AMP배지에서 생육하는 500여종의 방선균을 분리하였으며, 그 중 전배양배지(GMY 배지)에서 가장 뛰어난 항암활성능을 소유한 균주를 불리하고 배양하여 세포외 항암성물질을 분리하여 이 물질을 완전정제하고 MTT 정량분석을 실시하여 암세포에 대한 세포독성검사를 실시하였다. 1. 정제는 배양한 균체를 완전히 제거하고 동량의 ethylacetate를 처리하여 배양액중의 항암성분을 추출하고 무수 magnesium sulfate로 건조 후 농축, ethanol로 용해, 10배량의 water을 첨가하여 $4^{\circ}C$에서 overnight 시킨 후 추출액을 원심분리(12,000 rpm, 30분)하여 methylene chloride로 용해시켜 silica gel 60 column(${\phi}35{\times}600mm$, Merck Co.), methylene chloride-ethanol(96:4) 용매로 용출하고 sephadex LH-20 column(${\phi}15{\times}300mm$, Pharmacia LKB)에서 100% methanol로 용출하여 HPLC(Waters, ODS)를 이용하여 최종적으로 완전 정제하였다. 2. Compound는 Gram(+) 세균(6균주), Gram(-) 세균(11균주), 효모(2균주), 곰팡이(1균주)에 대하여 항균 효과를 나타내었다. 3. 완전 정제된 물질에 대한 항암효과를 측정한 결과, 모든 실험 암세포에 대해서 뛰어난 항암효과를 나타내었으며 protein성 물질은 아닌 것으로 추정되었다.

• Applied Microscopy
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• 제20권1호
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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.