• 제목/요약/키워드: cytochemical analysis

검색결과 13건 처리시간 0.025초

Effects of Wax Gourd Extracts on Adipocyte Differentiation and Uncoupling Protein Genes(Ucps) Expression in 3T3-Ll Preadipocytes

  • Kang, Keun-Jee;Kwon, So-Young
    • Nutritional Sciences
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    • 제6권3호
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    • pp.148-154
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    • 2003
  • Although various raw plant materials have been demonstrated to exert anti-obesity effects to a greater or lesser extent in both humans and animals when they are used to supplement the diet, it has not been shown extensively that they influence adipocyte cell differentiation involving lipid metabolic gene expressions. Using a well-established 3T3-L1 preadipocyte differentiation system, we decided to look into molecular and cellular event occurring during adipocyte differentiation when raw plant materials aye included in the process, in an effort to demonstrate the potential use of a screening system to define the functions of traditionally well-known materials. To these ends, the effects of ethanol (EtOH) or EtOH/distilled water (DW) extracts of Wax Gourd were examined using cytochemical and molecular analyses to determine whether components of the extracts modulate adipocyte differentiation of 3T3-Ll preadipocytes in vitro. The cytochemical results demonstrated that EtOH or EtOH/DW extracts did not affect lipid accumulation and cell proliferation, although the degree of lipid accumulation was influenced slightly depending on the extract. EtOH extract was highly effective in apoptotic induction during differentiation of 3T3-Ll preadipocytes (p<0.05). Reverse transcription-polymerase chain reaction (RT-PCR) analysis of lipoprotein lipase (LPL), Uncoupling protein (Ucp) 2, 3 and 4 also showed that while LPL expression was not influenced, Ucp2, 3 and 4 were up regulated in the EtOH extract-treated group and down regulated in the EtOH/DW extract-treated group. These changes in gene expressions suggest that the components in different fractions of Wax Gourd extracts may modulate lipid metabolism by either direct or indirect action. Taking these results together, it was concluded that molecular and cellular analyses of adipocyte differentiation involving lipid metabolic genes should facilitate understanding of cellular events occurring during adipocyte differentiation. Furthermore, the experimental scheme and analytical methods used in this study should provide a screening system for the functional study of raw plant materials in obesity research.

효모 세포의 Tripolyphosphatase와 Polyphosphatase 활성도 및 Volutin 과립의 세포학적 관찰 (Cytochemical Observation of Volutin Granules and Activities of Tripolyphosphatase and Polyphosphatase in Saccharomyces uvarum)

  • 이기성;최영길
    • 한국균학회지
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    • 제13권3호
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    • pp.141-148
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    • 1985
  • 이 연구는 효모 세포를 catabolic repression과 derepression시켰을 때, poly-P 합성 및 분해에 직접 관련된 tripoly-Pase와 poly-Pase의 활성도 동태를 조사하여 세포내 인산 대사 조절양상을 해석하고자 하였다. 또 TLC 방법을 통하여 무기 폴리인산을 분석하고, 배양시기 및 인산 첨가배양에 따른 무기폴리인산의 세포내 축적 상태, 세포내 위치, 전환과정을 조사하기 위해 염색을 통한 volutin 과립의 세포학적 관찰을 행하였다. Tripoly-Pase 활성도는 catabolic repression시킨 세포에서 derepression된 세포에 비하여 6배 이상 증가하였으며, poly-Pase 활성도는 산 불용성 무기폴리인산인 poly-P-"B"가 최대 축적량을 나타날 시기에 최대의 활성도를 보였다. Linear poly-P의 분석에서는 tripoly-P가 주로 검출되었고 n>8이상인 poly-P가 다량 검출되었다. catabolic derepression시킨 세포에서 volutin과립의 형성을 세포학적으로 관찰하였을 때, 세포벽에 존재하는 산 불용성 무기 폴리인산이 우선적으로 합성되었으며, 배양함에 따라 세포질, 핵 또는 액포로 전이되었다.

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비행운동 제한에 따른 비둘기 심근세포의 퇴행성 변화 (Degenerative Changes of Myocytes Induced by Restriction of Flight in Doves)

  • 문혜정;이용덕;박원학
    • 대한의생명과학회지
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    • 제6권4호
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    • pp.269-279
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    • 2000
  • 격렬한 운동유형으로 분류되는 비행이 습성화 되어 있는 조류를 대상으로, 인위적으로 비행을 제한시켰을 떼 나타나는 심근의 퇴행성 변화를 살펴봄으로써, 운동량 감소가 심근에 미치는 영향에 대하여 검토하고자 하였다. 서식환경의 제한을 위해 자연상태에서 서식하는 비둘기를 1 $m^3$ 공간에 2달 동안 서식시킨 군 (confined type)과 자연상태의 군 (wild type)으로 나누어 심근의 변화양상을 미세구조적, 세포화학적 방법 및 입체해석학적 방법으로 분석하였다. 체중은 confined type에서 증가되어 상대적으로 심장의 무게는 감소되었고, 흉근의 무게는 6.5% 감소 (p<0.05) 되었다. Confined type은 wild type과 비교해 볼 때, 미세구조적으로 더 적은 빈도의 심근의 파동형 변화와 지방소적의 침적이 관찰되었고, 근절의 길이는 길게 관찰되었다. Cytochrome oxidase의 활성을 알아보기 위해 실시한 세포 화학적 반응 결과, confined type에서 활성이 15% 감소되었다. 또한 입체해석학적 분석 결과, confined type의 관원섬유 체적밀도는 높게 나타났으나, 근형질내세망의 체적밀도와 사립체 내막의 표면밀도는 각각 23%, 25% 낮게 (p<0.05, p<0.01) 나타났다. 사립체의 체적밀도는 confined type에서 낮게 나타난 반면, 수적 밀도는 wild type과 비교하여 17% 높게 (p<0.05) 나타났다. 이러한 결과를 종합해보면 비행이라는 고에너지를 요구하는 격렬한 운동인 비행이 습성화 되어 있는 비둘기에서 비행을 제한시키면, 심근은 운동량의 감소에 따른 퇴행성 변화를 겪게 되므로, 운동의 지속성 여부는 심근기능유지에 매우 중요한 요소로 판단된다.

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Aspergillus nidulans 색소결핍 억제돌연변이주의 세포벽 미세구조 (Ultrastructure of Cell Wall in the Suppressor Mutant of Null Pigmentation (SU-NPG) of Aspergillus nidulans)

  • 정윤신
    • 생명과학회지
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    • 제14권1호
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    • pp.45-50
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    • 2004
  • A. nidulans의 색소결핍 억제돌연변이주(Suppressor mutant of the null pigmentation; SU-NPG)의 세포벽 구조가 색소형성 및 균사분지에 미치는 영향을 조사하여 다음의 결과를 얻었다. SEM으로 세포 표면구조를 관찰한 결과, 완전배지에서 배양된 SU-NPG의 분생포자벽은 최외각 층이 6일 째부터 박리되었으며, 균사생장이 저조하였다 TEM으로 관찰한 결과, 균사 세포벽은 매우 두꺼워졌다. 이러한 전자현미경관찰 결과는 이들 돌연변이주의 세포벽 구조가 NPG 및 야생형의 그것과는 다르다는 것을 시사한다. 균사에 탄수화물 염색을 한 후, 세포화학적 미세구조를 관찰한 결과, 분생포자벽은 C1, C2, C3와 C4 층으로 되어 있었고, 균사 세포벽은 H1, H2, H3와 H4층으로 되어 있었다. 이러한 C3층과 H3층은 세포분화 시 색소형성 및 균사 분지형성에 필수적인 세포벽 구조임 을 시사한다. SU-NPG는 FGSC4에서와는 다르게 비정상적인 균사생장을 하였지만, 색소결핍 돌연변이를 억제하는데, 이는 SU-NPG의 균사 세포벽에는 H3층이 형성되었으며, 세포벽 형성과정은 정상적으로 진행되기 때문으로 사료된다.

Isolated Spinal Neurocysticercosis : Unusual Ocular Presentation Mimicking Pseudotumor Cerebri

  • Seo, Jong-Hun;Seo, Hong-Ju;Kim, Seok-Won;Shin, Ho
    • Journal of Korean Neurosurgical Society
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    • 제49권5호
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    • pp.296-298
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    • 2011
  • Spinal intradural cysticercosis is a rare manifestation of neurocysticercosis. We report a unique patient who showed visual symptoms and normal imaging of the brain caused by isolated spinal neurocysticercosis. A 59-year-old male patient was admitted to the emergency unit with a history of severe headache and progressive blurred vision. Brain computed tomographic scanning and magnetic resonance imaging showed normal cerebral anatomy without hydrocephalus. The fundoscopic evaluation by an ophthalmologist showed bilateral papilledema. Perimetry studies revealed visual field defects in both eyes. With the diagnosis of pseudotumor cerebri, a lumbar tap was attempted; however, we could not drain the cerebrospinal fluid in spite of appropriate attempts. Lumbar magnetic resonance imaging revealed multilevel intraspinal lesions that were confirmed histologically to be neurocysticercosis. An intraoperative lumbar puncture revealed an increased opening pressure and cytochemical analysis showed elevated cerebrospinal fluid protein level. The headache resolved immediately after surgery. However, the visual symptoms remained and recovered only marginally despite antihelminthic medications after six months of operation.

Relationship between the nucleolar cycle and chromatoid body formation in the spermatogenesis of $Phrynops$ $geoffroanus$ (Reptilia Testudines)

  • Peruquetti, Rita L.;Taboga, Sebastiao R.;Cabral, Silvia R.;De Oliveira, Classius;Azeredo-Oliveira, Maria T.
    • Animal cells and systems
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    • 제16권2호
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    • pp.104-113
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    • 2012
  • The nucleolus is a distinct nuclear territory involved in the compartmentalization of nuclear functions. There is some evidence of a relationship between nuclear fragmentation during spermatogenesis and chromatoid body (CB) formation. The CB is a typical cytoplasmic organelle of haploid germ cells, and is involved in RNA and protein accumulation for later germ-cell differentiation. The goal of this study was to qualitatively and quantitatively describe the nucleolar cycle during the spermatogenesis of $Phrynops$ $geoffroanus$ (Reptilia Testudines), and compare this nucleolar fragmentation with CB formation in this species through the use of cytochemical and ultrastructural analysis. Qualitative analysis showed a fragmentation of the nuclear material after pachytene of the first meiotic division in the primary spermatocytes. Quantitative analysis of the nucleolar cycle revealed a significant difference in the number of nucleoli and in the size of the nucleolus between spermatogonia and early spermatids. Using ultrastructural analysis, we recorded the beginning of the CB formation process in the cytoplasm of primary spermatocytes at the same time as when nuclear fragmentation occurs. In the cytoplasm of primary spermatocytes, the CB was observed in association with mitochondrial aggregates and the Golgi complex. In the cytoplasm of early spermatids, the CB was observed in association with lipid droplets. In conclusion, our data show that the nucleolus plays a role in the CB formation process. During spermatogenesis of $P.$ $geoffroanus$, the CB is involved in some important biological processes, including acrosome formation and mitochondrial migration to the spermatozoon tail and middle piece region.

한국관박쥐 망막에서 글루타메이트 수용체의 분포 양상 (Distribution of Glutamate Receptors in the Retina of the Greater Horseshoe Bat (Rhinolophus ferrumequinum))

  • 권오주;전창진
    • 한국안광학회지
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    • 제19권3호
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    • pp.413-418
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    • 2014
  • 목적: 한국관박쥐 망막의 기능을 알아보기 위해서 흥분성 신경전달물질인 글루타메이트 수용체의 분포도를 분석하였다. 방법: 성체 한국관 박쥐의 망막을 $40{\mu}m$ 수직 절편 한 후 표준면역세포화학법을 이용하였다. 면역형광이미지는 Bio-Rad MRC 1024 공초점 현미경을 사용하여 얻었다. 결과: AMPA (GluR1-4), Kainate (GluR5-7, KA1-2), NMDA (1, 2A, 2B)는 내망상층과 외망상층에 주로 분포되어 있었다. KA1은 신경절세포층에도 많은 수의 수용체가 존재하였다. 결론: 한국관박쥐는 포유류망막에 있는 신경세포와 신경전달물질을 동일하게 가지고 있었다. 한국관박쥐도 기능적 망막을 가지고 있음을 제시한다.

Ultrastructure of the Cell Wall of a Null Pigmentation Mutant, npgA1, in Aspergillus nidulans

  • Chung, Yun-Shin;Kim, Jung-Mi;Han, Dong-Min;Chae, Keon-Sang;Jahng, Kwang-Yeop
    • Journal of Microbiology
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    • 제41권3호
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    • pp.224-231
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    • 2003
  • The null pigmentation mutant (npgA1) of Aspergillus nidulans was previously characterized by its production of no pigment at any stage of its life cycle, its reduction in hyphal branching, and its delay in the asexual spore development. The chemical composition of the cell wall was also altered in npgA1 mutants that became more sensitive to Novozyme 234$\^$TM/, which is possibly due to a structural defect in the cell wall. To investigate the effects of the cell wall structure on these pleiomorphic phenomena, we examined the ultrastructure of the cell wall in the npgA1 mutant (WX17). Scanning electron micrographs (SEM) showed that after being cultured for six days, the outermost layer of the conidial wall of WX17 peeled off. Although this phenotype suggested that the cell wall structure in WX17 may be modified, examination using TEM of the fine structure of cross-sectioned hyphal wall of WX17 did not show any differences from that of FGSC4. However, staining for carbohydrates of wall layers showed that the electron-translucent layer of the cell wall was missing in WX17. In addition, the outermost layer H1 of the hyphal wall was also absent in WX17. The ultrastructural observation and cytochemical analysis of cell walls suggested that the pigmentation defect in WX17 may be attributed to the lack of a layer in the cell wall.

Conjugated Linoleic Acid 이성체가 3T3-L1 지방전구세포 분화중 세포증식, 세포사멸 및 Ucp 유전자 발현에 미치는 영향 (The Effect of Conjugated Linoleic Acid Isomers on the Cell Proliferation, Apotosis and Expressions of Uncoupling Protein (Ucp) Genes during Differentiation of 3T3-L1 Preadipocytes)

  • 권소영;강금지
    • Journal of Nutrition and Health
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    • 제37권7호
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    • pp.533-539
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    • 2004
  • It has been reported that CLA decreases fat deposition in vivo and in vitro experiments. Among CLA isomers, c9t11 and t10c12 have been shown to exert active biological activities. For example, t10c12 reduces body weight and increases lean body mass, whereas, c9t11 has little effect on body fattness. However, the underlying molecular mechanism for the anti-obesity action of CLA isomers are not well understood. The purpose of this study was to examine the effects of t10c12 and c9t11 on lipid accumulation, cell proliferation, cell death and the expression levels of Ucp genes which are proposed as targets for anti-obesity in 3T3-L1 preadipocytes. Isomers of CLA at 50$\mu$M were added into preadipocyte differentiation medium for 3, 6 and 9days. Control cells received only the vehicle in the differentiation medium. Cytochemical analyses for lipid accumulation, cell proliferation and apotosis were carried out to compare lipidogenesis and cellular activity. RT-PCR analysis of GAPDH, Ucp 2,3 and 4 were also performed to find any modulatory effects of CLA isomers on the metabolic genes. Lipid accumulation indicated by Oil Red-O staining was inhibited in CLA isomers as compared to the control. T10c12 isomer showed less lipidogenesis than c9t11 did. A decrease occurred in CLA isomers as shown by BrdU incorporation. Apotosis has occured at higher level in t10c12 when compared to that of t9c11. Ucp 2, 3 and 4 genes were also upregulated in CLA isomers. T10c12 showed higher level of Ucp gene expressions than the c9t11 did. The biological activities of CLA isomers were also found to be different during differentiation of 3T3-L1 preadipocytes, suggesting that different isomers may be active in certain stage of lipidogenesis. The results indicate that both c9t11 and t10c12 CLA isomers decrease lipidogenesis, inhibit cell proliferation, increase cell death and upregulate in Ucp gene expressions during 3T3-L1 preadipocyte differentiation. T10c12 isomer was more effective than c9t11 in overall anti-obesity activity.

흰쥐 골격근의 노화성 위축에 대한 운동훈련의 영향 - II. 장기간에 걸친 체중부하 훈련의 영향 - (Effect of Exercise Training on Aging Atrophy in Rat Skeletal Muscle II. Effect of Long Term Weight-Training)

  • 박승한;박원학;이용덕;김정기
    • Applied Microscopy
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    • 제25권4호
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    • pp.26-51
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    • 1995
  • The present study was designed to examine effect of long term weight-training on aging atrophy in the rat skeletal muscle. Male rats of 8, 15, and 24 month old were used. Each age groups included control and weight-training for 5 months by using body press apparatus. The histo- and cytochemical, ultrastructural and stereological changes in aging skeletal muscles of the rat were observed in the present study. During the training period the body weight and muscular weight in all groups except the rectus femoris and the gastrocnemius in young age groups remained constant, but muscular weights were increased in the rectus femoris and the gastrocnemius muscles in young age groups. In trained rat, the volume density of muscle fiber type IIA and IIB were increased, but those of type IIC was decreased. Type I remained constant in 8 and 15 month old age groups, but reduced in the tibialis anterior and the gastrocnemius muscles in the 24 month old groups. Some histotological and ultrastructural changes associated with age were found: numerical increase of cytiplasmic vacuoles, lysosomes, lipofuscins, and irregularity of myofibrils. At 24 month old groups some unusual formation of contraction band and muscle splitting were observed. After weight-training, ultrastructural degenerative changes occured in the type I muscle fiber, such as splitting of muscle fiber, disorganization of myofilaments, swelling of mitochondria, accumulation of many lipid droplets, appearance of many lysosomes and residual bodies and necrotic fibers, in the old age groups. But, in the type II muscle fibers hypertrophy of muscle fiber appeared without any noticible damage as the type I. The activities of $Mg^{++}$ -ATPase decreased with age and this enzyme activities in the trained rat were significantly decreased with age. Activities of the acid phosphatase were increased with age and significantly in the trained rat. In stereological analysis, volume density of the myofibrils and the tubular system were increased, on the other hand there mitochondrial capacity was decreased. These experimental results suggested that old rats are not susceptible to be affected by weight-training as young rats, and that physical capacity of the rats must be considered when old rats are exercised for training.

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