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Derivation of Neural Precursor Cells from Human Embryonic Stem Cells

  • Kim Sehee;Hong Ji Young;Joo So Yeon;Kim Jae Hwan;Moon Shin Yong;Yoon Hyun Soo;Kim Doo Han;Chung Hyung Min;Choi Seong-Jun
    • Reproductive and Developmental Biology
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    • v.28 no.4
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    • pp.247-252
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    • 2004
  • Human embryonic stem (ES) cells are derived from the inner cell mass of the preimplantation embryo. Human ES cells have the capacity to differentiate into various types of cells in the body. Human ES cells are indefinite source of cells for cell therapy in various degenerative disorders including neuronal disorders. Directed differentiation of human ES cells is a prerequisite for their clinical application. The objective of this study is to develop the culture condition for the derivation of neural precursor cells from human ES cells. Neural precursor cells were derived from human ES cells in a stepwise culture condition. Neural precursor cells in the form of neural rosette structures developed into neurospheres when cultured in suspension. Suspension culture of neurospheres has been maintained over 4 months. Expressions of nestin, soxl, sox2, pax3 and pax6 transcripts were upregulated during differentiation into neural precursor cells by RT-PCR analysis. In contrast, expression of oct4 was dramatically downregulated in neural precursor cells. Immunocytochemical analyses of neural precursor cells demonstrated expression of nestin and SOX1. When induced to differentiate on an adhesive substrate, neuro-spheres were able to differentiate into three lineages of neural systems, including neurons, astrocytes and oligo-dendrocytes. Transcripts of sox1 and pax6 were downregulated during differentiation of neural precursor cells into neurons. In contrast, expression of map2ab was elevated in the differentiated cells, relative to those in neural precursor cells. Neurons derived from neural precursor cells expressed NCAM, Tuj1, MAP2ab, NeuN and NF200 in immunocytochemical analyses. Presence of astrocytes was confirmed by expression of GFAP immuno-cytochemically. Oligodendrocytes were also observed by positive immuno-reactivities against oligodendrocyte marker O1. Results of this study demonstrate that a stepwise culture condition is developed for the derivation of neural precursor cells from human ES cells.

Electron-microscopic studies on fine structure and enzyme activity in the axenic and conventional strains of Entamoeba histolytica (이질아메바(Entamoeba histolytica)의 미세구조 및 효소활성에 관한 전자현미경적 연구)

  • Yong, Tae-Sun;Jeong, Pyeong-Rim;Lee, Geun-Tae
    • Parasites, Hosts and Diseases
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    • v.23 no.2
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    • pp.269-284
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    • 1985
  • The metabolism of Entamoeba histolytica would be affected by various environmental factors, and alteration of the environment was known to afEect the fine structure of 5. histolytica. The present study was designed electronmicroscopically to investigate the ultrastructure and enzyme activities in the aEonic and conventional strains of 5. histolytica. The trophozoites of axenically cultivated HK-9 strain and conventional YS-27 and YS-49 strains of 5. histolytica were collected and liKed with 4% paraformaldehyde/0.1M cacodylate buffier(pH 74), After washing them by centrifugation, 1% warm agar was added in the sediment. Solidified agar with the trophozoites was cut into $lmm^3$ cubes, and incubated in the various substrates to observe enzyme activities. Then, the specimen was post-fixed with 3% glutaraldehyde/0.1M cacodylate buffer (PH 7.4) and 1% osmium tetroBide/0.1M cacodylate buffier (pH 7.4) , dehydrated in ascending ethanol series and embedded in epoxy resin. These were sectioned on an ultramicrotome and observed with a transmission electronmicroscope. The procedures for the observation of the fine structure were same as the above, except for the incubation in the substrate. The sections were stained with uranyl acetate and lead citrate. For the observation of the surface of the amoebae, scanning-electronmicroscopy was carried out. The results obtained in the present study are summarized as follows: 1. The fuzzy coat around double-layered plasma membrane of 5. histolytica was more irregularly and densely distributed in the conventional strains (YS-27, YS-49 strains) than in the axonic strain (HK-9 strain). 2. The endosomes, button bodies and chromatin material were surrounded by a double-layered nuclear membrane having scattered nuclear fores. The paranuclear body, mono- or double-layered vacuoles, vacuolar membrane whorls, rosette-like cylindrical bodies, aggregation of cylindrical bodies and helical bodies were found in the cytoplasm of the amoebae. Helical bodies and glycogen granules were generally abundant, while a few smooth endoplasmic reticula were observed in the cytoplasm. 3. Alkaline phosphatase activity was mainly demonstrated in the plasma membrane, limiting membranes of vacuoles and smooth endoplasmic reticula. ATPase activity was observed in the nucleus, limiting membranes of vacuoles and vacuolar membrane whorls. 4. Acid phosphatase activity was commonly demonstrated in the limiting membranes an contents of vacuoles, Iysosome-like organelles, plasma membrane and the button bodies in the nucleus. The activity was more weakly demonstrated in the HK-9 strain than in the other conventional strains of 5. histolytica. No peroBidase activity was observed in the amoeba strains employed in the present study. 5. With a scanning electron-microscope, no distinct structural differences were observed between the amoeba strains. All the trophozoite forms of the amoebae showed crater-like depressions and rugged features on the outer surface.

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Arabidopsis thaliana의 Ethylene Triple Response Mutant에서 에틸렌 생합성 과정의 생리 생화학적 특성

  • 이준승
    • Journal of Plant Biology
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    • v.39 no.1
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    • pp.31-40
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    • 1996
  • The physiological and biochemical characterizations of the ethylene-related mutants in Arabidopsis thaliana - ethylene overproducing mutant (eto1-l) and ethylene insensitive mutants (etrl-3, ein2-l) - were detailed in this studies. Two or three week.old mature rosette leaves (before bolting) were used as the plant materials. Ethylene productions of eto1-l, etrl-3, and ein2-l mutants were about 200%, 400%, and 450% compared to that of wild type, respectively. ACC synthase and ACC oxidase activities of eto1-l mutant were similar to those of wild type. ACC content and ACC N-malonyltransferase activity, however, were 4.5 times and 3 times higher than those of wild type, respectively. SAM synthetase activity increased by 50% in eto1-l mutant plant. These results indicated that the alteration in the eto1-l mutant occured before the step of the conversion of SAM to ACe. In etrl-3 and ein2-l mutants, ACC synthase activities increased, but ACC oxidase activities decreased. ACC content and ACC N-malonyltransfcrase activity were 2 times higher than those of wild type. SAM synthetase activity in etrl-3 is similar to those of wild type, while it increased by 73% in ein2-l. These results showed that the block in ethylene action affected the autoregulation of ethylene biosynthesis, so that ACC synthase activity was not autoinhibited and ACC oxidase activity was not auto stimulated by ethylene. When the leaf tissues were used for in vitro kinase assay, a cytosolic protein (approximately 36 kDa) was phosphorylated only in eto1-l and ein2-l mutants.utants.

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The Experimental Study of the Effects of Sungyangikgibuja'ang and Kwangyebujalijungt' and of Soŭm-In on Yang-Insufficient Syndrome (소음인(少陰人) 승양익기부자탕(升陽益氣附子湯)과 관계부자리중탕(官桂附子理中湯)이 양허증(陽虛證)에 미치는 영향(影響)에 관(關)한 실험적(實驗的) 연구(硏究))

  • Jeon, Jin Sang
    • Journal of Sasang Constitutional Medicine
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    • v.1 no.1
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    • pp.87-112
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    • 1989
  • In order to investigate experimentally the effects of Sung-yangikgibujat'ang (SIT) and Kwangyebujalijungt'ang (KBT) on Yang-insufficient syndrome (陽虛證) induced by Hydrocortisone acetate (H.A.) in experimental animals (Mice and Rat), the author experimented various activities. Delayed type hypersensitivity (DTH), Rosette forming cells (RFC), hemagglutinin (HA) titers, Hemolysin (HL) titers, Body weight, Whole blood viscosity, Plasma Viscosity, Hematocrit, RBC, Albumin, Total protein, Triglyceride, cholesterol and Glucose were measured. The results summerized as follows; 1. In DTH and RFC all the experimental groups were increased significantly in comparison to the H.A.-treated group. 2. In HA titers SIT_treated group were increased significantly and KBT-treated group showed increasing tendancy, but showed no significance. 3. In HL titers all the experimental groups showed increasing tendancy, but showed no significance. 4. In body weight all the experimental groups showed increasing tendancy, but showed no significance. 5. In whole blood viscosity all the experimental groups were decreased significantly in comparison to the H.A.-treated group. 5. In whole blood viscosity all the experimental groups were decreased significantly in comparison to the H.A.-treated group. 6. In plasma viscosity KBT-treated group were decreased significantly and SIT-treated group showed decreasing tendancy, but showed no significance. 7. In Hematocrit SIT-treated group were decreased significantly and KBT-treated group showed decreasing tendancy, but showed no significance. 8. In RBC, albumin and cholesterol all the experimental groups were decreased significantly in comparison to the H.A.-treated group. 9. In total protein and triglyceride KBT-treated group were decreased significantly and SIT-treated group showed decreasing tendancy, but showed no significance. 10. In Glucose SIT-treated group were decreased significantly and KBT-treated group showed decreasing tendancy, but showed no significance. From above findings, it has been demonstrated that Sungyangikgibujat'ang and Kwangyebujalijungt'ang seem to produce the effectiveness on the recovery from depression of the cell-mediated immune response, blood circulation and energy metabolic rate, induced by Hydrocortisone acetate, and in the humoral immune response Sungyangikgibujat'ang have the effectiveness on the recovery, and in cellular component of blood Sungyangikgibujat'ang was more effective than Kwangyebujalijungt' ang, and in plasma of blood Kwangyebujalijungt'ang was more effective than Sungyangikgibujat'ang. Therefore it is suggested that Sungyangikgibujat'ang and Kwangyebujalijungt'ang have the effectiveness on the recovery from Yang-insufficient syndrome more or less.

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Germanium-Fortified Yeast Activates Macrophage, NK Cells and B Cells and Inhibits Tumor Progression in Mice. (게르마늄 강화효모의 마우스에서의 암세포 억제 및 대식세포, NK 세포, B 세포의 활성화에 관한 연구)

  • Baek, Dae-Heoun;Jung, Jin-Wook;Sohn, Tsang-Uk;Kang, Jong-Koo
    • Microbiology and Biotechnology Letters
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    • v.35 no.2
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    • pp.118-127
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    • 2007
  • Germanium-fortified yeast (GY) is a organic germanium-fortified yeast with potent immune modulating activities including anti-inflammatory effect. Through cell line studies, we observed that GY can modulate the diverse immune activity but little evidence was provided on the mechanism of GY in modulating immune activities in other higher animals. In this study, we investigated the effect of GY on modulation of immune function in mice. GY was administered in normal mice or tumor-bearing mice and then effect of GY on modulation of host immune system was analyzed by using ex vivo isolated macrophages, B cells, NK cells. Admistration of GY in mice induced macrophage activation thereby increased effector function of macrophage such as increased phagocytosis, chemotaxis, adherence, $O_2-release$, NO, $TNF-{\alpha}$ production. In addition, GY administration Increased B lymphocyte activation and plaque forming cells. Furthermore, GY administration increased NK-cell mediated cytotoxicity. Furthermore, GY administration suppressed progression of tumor in mice by increasing $TNF-{\alpha}$ production and effector function of NK cells. Our results showed that GY has a potent immunostimulatory function in vivo mice model. Proper modulation and administration of GY in human could be helpful to maintaining immunological homeostasis by modulating host immune system.

The distributional characteristics of the major dissolved artificial radionuclides in the adjacent seas of Korea(I : Yellow Sea) (우리나라 주변해역 주요 인공방사성 핵종 분포 특성 (I: 황해))

  • Chung Chang Soo;Kim Young ill;Moon Deok Soo;Kim Suk Hyun;Park Jun Kun;Seo Seung Mo;Hong Gi Hoon
    • Journal of the Korean Society for Marine Environment & Energy
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    • v.4 no.1
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    • pp.3-13
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    • 2001
  • Dissolved /sup 137/Cs, /sup 239.240/Pu, /sup 238/Pu and /sup 90/Sr contents in winter and spring of the Yellow Sea were determined to describe the distribution of artificial radionuclides. Surface water samples (100 liter) were collected by using a submerged pump, and subsurface samples (>10m depth) were collected using a 10L Niskin water sampler mounted to the Rosette sampler. The levels in the surface water ranged between 1.78~3.38 mBq kg/sup -1/ for /sup 137/Cs, 2.17~13.35 μBq kg/sup -1/ for /sup 239,240/Pu, and 1.97~3.96 mBq kg/sup -1/ for /sup 90/Sr, respectively. In particular, the concentration of /sup 239.240/Pu were 1/10 of those in the vicinity of Changjiang estuary (61~83 μBq kg/sup -1/). The difference of /sup 238.240/Pu concentration between surface and bottom water was <3.0 μBq kg/sup -1/in the Yellow Sea. It suggests that in the Yellow Sea which has shallow and high suspended sediments, /sup 239.240/Pu is preferentially removed from the water columm. The water column inventory of /sup 239.240/Pu in the Yellow Sea constitute about 0.7~0.9 % of the estimated fallout input to the area. The activity ratios of /sup 239.240/Pu//sup 137/Cs and /sup 137/Cs//sup 90/Sr ranged between 0.001~0.005, 0.79~1.65, respectively, and similar to those of open ocean which global fallout is the only source of artificial radionuclides. Therefore, it suggests that most of these artificial radionuclides in the Yellow Sea may be controlled by the atmospheric input.

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Peripheral Neuroblastoma of the Ulnar Nerve : Diagnosis by Fine Needle Aspiration Cytology (척골신경에 발생한 말초성 신경아세포종 -세침흡인 세포검사로 진단된 1례 보고-)

  • Chu, Young-Chae;Kim, Joon-Mee
    • The Korean Journal of Cytopathology
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    • v.4 no.1
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    • pp.45-51
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    • 1993
  • A 30-year-old woman who was diagnosed as peripheral neuroblastoma by fine needle aspiration of a soft mass of the right upper arm is described. She presented a slowly growing, soft mass of the right upper arm for 1 month. The right humerus revealed no abnormal finding on X-ray. Ultrasonogram of the right upper arm revealed a well demarcated, smooth marginated solid mass without invasion of adjacent structures. Fine needle aspiration was done under the impression of soft tissue tumor with undetermined biologic behavior. The aspirates were highly cellular and the tumor cells were dispersed both singly and in clusters of varying size. The clusters occasionally showed a central capillary core and rosette-like structures. The tumor cells were small in size and had a small to medium amount of cytoplasm. Some of them revealed slender cytoplasmic processes. The nuclei showed distinct nuclear membranes, finely clumped chromatin and small conspicuous nucleoli. Cellular pleomorphism or mitotic figure was not definite. These cytologic findings were interpreted as a malignant, non-lymphomatous small round cell tumor, most likely representing peripheral neuroblastoma or Ewing's sarcoma. Final diagnosis was confirmed by simple excision as peripheral neuroblastoma.

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First Report of Pink Rot of Potato (Solanum tuberosum) Caused by Phytophthora erythroseptica in Korea (Phytophthora erythroseptica에 의한 감자 홍색부패병 발생)

  • Ryu, Kyoung-Yul;Kim, Jeom-Soon;Kim, Jong-Tae;Hahm, Young-Il
    • Research in Plant Disease
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    • v.9 no.1
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    • pp.32-35
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    • 2003
  • Pink rot of potato (Solanum tuberosum L.) occurred at Pyeongchang in Gangwon and at Bosung in Junnam province since 1999. The disease incidence in the surveyed areas was about 5% of harvested potatoes in 2002. Affected tubers showed a dull brown appearance and the lenticels and eyes on tubers turned dark brown. The cut surface of the diseased tuber colored faint pink and the entire surface of the diseased tuber becomes deep salmon pink within 30 min. The pathogen isolated from the diseased tubers was identified as Phytophthora erythroseptica based on morphological and cultural characteristics. Mycelial mat was fairly fluffy, rosette or stellate patterns and rounded or angular hyphal swellings were farmed in water, Temperature for mycelial growth was ranged from 5 to 3$0^{\circ}C$ and optimal temperature was $25^{\circ}C$. Non-papillate sporangia were persistent on stalk and ellipsoid, ovoid, obpyriform or distorted in shape, often with a constriction distal in the middle. Size of sporangia was 41.3~69.6$\times$26.8~47.4 (av, 55.5$\times$37.1) ${\mu}{\textrm}{m}$. Sexuality of Phytophthora erythroseptica was homothallic. Oogonia were 30~46 ${\mu}{\textrm}{m}$ in diameter and oospores were 28~35 ${\mu}{\textrm}{m}$ in diameter, Elongated or cyclindrical antheridia were all amphigynous. This is the first report on potato pink rot caused by Phytophthora erythroseptica in Korea.

Characteristics of Growth and Development of Cuttings and Rooted Cuttings affected by Natural Low Temperature in Chrysanthemum 'Baekma' (국화 '백마'의 자연저온을 받은 삽수 및 묘의 생육 특성)

  • Choi, Seong Youl;Lim, Jin Hee;Park, Sang Kun;Kil, Mi Jung
    • FLOWER RESEARCH JOURNAL
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    • v.19 no.2
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    • pp.96-102
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    • 2011
  • The objective of study was carried out to find a proper entrance date for breaking dormancy depending on cutting and entrance date into greenhouse investigating plant growth and flowering characteristics of chrysanthemum 'Baekma'. Days to visible bud formation and days to flowering were increased as cutting date was delayed. The flowering rate of cutting on September 18 was 100% while cutting on October 30 did not induce flower bud formation. Flower characteristics were surveyed after 'Baekma' and 'Jinba' rooted cuttings were planted on September 10. Branching formation rate of 'Baekma' was gradually increased as entrance date was delayed while that of 'Jinba' was about 70-80% regardless of entrance date. Lethality of 'Baekma' moving into greenhouse on December 20 was 1.5% by cold injury. 'Jinba' started to die on November 10 by cold injury and lethality of 'Jinba' moving into greenhouse on December 20 was 21.7%. Thus, lethality of 'Jinba' was about 5-14 times higher than that of 'Baekma'. Days to visible bud formation and days to flowering were decreased as entrance date was delayed in chrysanthemum 'Baekma'. Days to visible bud formation of entrance date on November 10 and December 10 were 67.9 and 50.3, respectively. On the other hand, that of 'Jinba' was increased until entrance date on December 10 and decreased on December 20. Based on these results, it was suggested that dormancy of 'Baekma' was started at late September and completed at late October.

In vitro CaCO3 Crystallization at Room Temperature and Atmospheric Pressure Using Recombinant Proteins GRP_BA and GG1234 (재조합단백질 GRP_BA 및 GG1234를 이용한, 상온상압조건에서의 In vitro 탄산칼슘 결정화)

  • Son, Chaeyeon;Song, Wooho;Choi, Hyunsuk;Choi, Yoo Seong
    • Korean Chemical Engineering Research
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    • v.57 no.2
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    • pp.205-209
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    • 2019
  • The exquisite structure and attractive biological properties of biominerals have great potential and increased interest for use in a wide range of medical and industrial applications. Calcium carbonate biomineralization, mainly controlled by shell matrix proteins, has been used as a representative model to understand the biomineralization mechanism. In this study, in vitro calcium carbonate crystallization was carried out under room temperature and atmospheric pressure using recombinant shell matrix protein GRP_BA and artificial shell matrix protein GG1234. Both proteins inhibited the growth of typical rhombohedral calcite crystals in the calcium carbonate crystallization using $CaCl_2$ solution and $(NH_4)_2CO_3$ vapor, and spherulitic calcite crystals with rosette-like structures were synthesized in both the presence of GRP_BA and GG1234. These results might be caused by the properties of block-like domain structure and intrinsically disordered proteins. We expect that this study can contribute to enhance understanding of the calcium carbonate biomineralization controlled by shell matrix proteins.