• Title/Summary/Keyword: Nuclei

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Optimized Methods for the Isolation of Arabidopsis Female Central Cells and Their Nuclei

  • Park, Kyunghyuk;Frost, Jennifer M.;Adair, Adam James;Kim, Dong Min;Yun, Hyein;Brooks, Janie S.;Fischer, Robert L.;Choi, Yeonhee
    • Molecules and Cells
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    • v.39 no.10
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    • pp.768-775
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    • 2016
  • The Arabidopsis female gametophyte contains seven cells with eight haploid nuclei buried within layers of sporophytic tissue. Following double fertilization, the egg and central cells of the gametophyte develop into the embryo and endosperm of the seed, respectively. The epigenetic status of the central cell has long presented an enigma due both to its inaccessibility, and the fascinating epigenome of the endosperm, thought to have been inherited from the central cell following activity of the DEMETER demethylase enzyme, prior to fertilization. Here, we present for the first time, a method to isolate pure populations of Arabidopsis central cell nuclei. Utilizing a protocol designed to isolate leaf mesophyll protoplasts, we systematically optimized each step in order to efficiently separate central cells from the female gametophyte. We use initial manual pistil dissection followed by the derivation of central cell protoplasts, during which process the central cell emerges from the micropylar pole of the embryo sac. Then, we use a modified version of the Isolation of Nuclei TAgged in specific Cell Types (INTACT) protocol to purify central cell nuclei, resulting in a purity of 75-90% and a yield sufficient to undertake downstream molecular analyses. We find that the process is highly dependent on the health of the original plant tissue used, and the efficiency of protoplasting solution infiltration into the gametophyte. By isolating pure central cell populations, we have enabled elucidation of the physiology of this rare cell type, which in the future will provide novel insights into Arabidopsis reproduction.

The Government Approach to the Eipty Nucleus (지배음운론에서 본 'ㅡ'모음)

  • Heo Yong
    • MALSORI
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    • no.19_20
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    • pp.58-87
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    • 1990
  • According to Government Phonology, at 1 phonological positions save the domain's head must be licensed in order to appear in the syllable structure. A non-nuclear head is licensed by the following nucleus, and the nuclei with phonetic content are licensed through government by the nuclear head of the domain at the level of the nuclear projection. Therefore, in the theory of Government Phonology it is claimed that words always end with a nucleus. With regard to the licensing of empty nuclei, Kaye(1990a) proposes the 'Empty Category Principle' and its sub-theory of 'Projection Government'. Government Phonology claims that a nucleus which dominates a vowel that regularly undergoes elision in certain contexts is underlyingly empty. This underlying empty nucleus is not manifested phonetically when it is properly governed by an unlicensed(i, e, a nucleus filled with a full vowel). It is when proper government fails to apply, that the empty nucleus is phonetically Interpreted. The purpose of this paper is to present a principled account of the process of $[i]{\Leftrightarrow}{\emptyset}$ alternation in Korean. Following Kaye's proposal, we assume that [i] of Korean is underlyingly empty. This position is pronounced as [i] if it is unlicensed, and is not phonetically realized if is licensed. Empty nuclei ape devided into two categories: domain-internal and domain-final. Firstly, we consider the question why Korean has little word ending with [i]. As for this, ECP states that domain-final empty nuclei are not pronounced if the language licenses domain-final empty nuclei. Whether a final empty nucleus may occur in the structure is parametric variation. This property is seen from the fact that words may appear to end in consonants in this language. Since Korean abounds with words ending in a consonant, it licenses domain-final empty nuclei. Therefore, it is quite natural that Korean has little word ending with [i]. Secondly, word-internal empty nuclei of Korean respect proper government and inter-onset government. That is, an empty nucleus in word-internal position will be pronounced with the vowel [i] if either proper government or inter-onset government fail to apply. Inter-onset government refers to the government established between two onsets across an empty nucleus. Thirdly, we consider words ending with [i], which seems to be exceptional to the final licensing. Host of them are. either mono-syllabic verbs(for instance, [s'i-] 'to write') or derived adjectives ending with [p'i] (for instance, [kip'i-] 'be happy'). As for the former, the 'inaccessibility for proper government' is applied because the empty nucleus appears in the first syllable. In latter case, domain-final empty nuclei are pronounced as [i] because of government-licensing. That is, final empty nucleus is pronounced to license the preceding onset dominating negatively charmed segments which empty nucleus of Korean cannot license.

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Role of Posterior Intralaminar Thalamic Nuclei in Acupuncture Analgesia in Rats (침진통(鍼鎭痛) 작용에 있어서 시상 후방 층판내핵(속방핵)의 역할)

  • Roh, Sik;Min, Byung-Il;Yoon, Sang-Hyub
    • The Journal of Korean Medicine
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    • v.20 no.3 s.39
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    • pp.36-44
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    • 1999
  • Objectives: It has been well known that electroacupuncture(EA) has an analgesic effect and there is a pain control system in the central nervous system(CNS). The pain control system is composed of three major nuclei, which are periaqueductal gray(PAG), raphe nuclei, and the pain inhibitory complex located in the spinal cord. It has been suggested that the analgesic effect of EA might be the result of activation of the pain control system in the CNS. However, there may be a possibility that other nuclei are also involved in this pain modulation. Thus, we investigated whether the posterior intralaminar thalamic nuclei (PTIN) are involved in the pain modulation. Methods: To measure the level of pain, the jaw opening reflex (JOR) was used as a pain index. The magnitude of JOR is estimated by averaging the area of 10 successive responses. JOR was evoked by tooth-pulp stimulation with bipolar electrode carrying stimulus with the following parameters: intensity ranging from 420uA to 680ulA, 0.3ms duration of square pulse, and 0.5 Hz. Hapkog($LI_4$) and Taechung ($LR_3$) were the chosen acupoints. The Hapkog point was stimulated ipsilaterally at 5V, 3 Hz, for 15min in total, and the Taechung was stimulated at 2-3 V, 3 Hz, and for a total of 15 or 30 minutes. Different intensities of stimulation were given the PITN; one was given at $300{\mu}A$ and the other was at 500uA. The position stimulated in these nuclei by Paxinos Atlas was AP; from bregma $-4.0{\sim}-4.3mm,\;L; 0.5{\sim}1.8mm,\;D;\;4.8{\sim}6.3mm$. Results: The Hapkog point had a significant analgesic effect (P<0.05). However, the Taechung point had no effect. Both types of stimulation in the PITN did not reveal any analgesic effects. Conclusions: From these results, it was suggested that the posterior intralaminar thalamic nuclei are not involved in the modulation of pain.

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Effect of Aflatoxin $B_1$ on Ultrastructural Changes of Biliary Epithelial Cells in Mice Experimentally Infected with Clonorchis sinensis (간흡충 감염 마우스에 있어서 환관 상피세포의 미세구조에 대한 Aflatoxin $B_1$의 영향)

  • 민홍기
    • Parasites, Hosts and Diseases
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    • v.25 no.2
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    • pp.99-109
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    • 1987
  • The present study was carried out to examine the effect of a calcinogen, aflatoxin B1 on the ultrastructural changes of ciliary epithelial cells in mice infected with Clonerchis sinensis. A total of 93 male albino mice(BALB/c strain) was divided into 3 groups; group I, treated with 1.0 ppm aflatoxin Bl for 12 weeks; group II, given 50 C. sinensis n;etacercariae, and group III, given 50 metacercariae and treated with 1.0 ppm aflatoxin Bl for 12 weeks. Three mice served for untreated-uninfected controls. From 4 weeks after the treatsment and/or in(ection, three mice from each group were sacrificed at 4 week intervals up to the 40th week, and their hepatobiliary tissues were prepared for transmission electron microscopy. The most prominent ultrastructural changes in group I were remarkable enlargement of nuclear size, separation of nucleolus, dispersed chromatin granules in nuclei and increased dense granules along the inner membrane of nuclei. In the cytoplasm there was slight proliferation of mitochondria and endoplasmic reticulum (ER) at earlier stage. At the 12th week separation of fibrillar and granular components of the nucleolus was a characteristic finding. As the time elapsed, epithelial cells showed fiattened-cuboidal form and a tendency of atrophy. Most of the nuclei were elongated and polygonal in shape. In group II the appearance of elaborate interwoven folds of lateral cytoplasm forming a labyrinth of interconnected intercellular space and variety in nuclear shape were the prominent fadings at earlier stage. The cytoplasm showed slight proliferation and dilatation of mitochondria and ER, and a small number of mucin droplets. In the basement membrane scanty fibrous cells were seen. With time, variety in nuclear shape, marked proliferation and dilatation of rough ER and some collagen fibrils were demonstrated. Other features of intracellular organelles and mucin droplets persisted. In group III cuboidal epithelial cells showed their remarkably enlarged and irregular nuclei, increased chromatin granules in the nuclei, separated nucleoli, proliferated and dilated rough ER. With time, sequestered mitochondria showed blob-like evaginations which lacked cristae and dense matrix, and were limited by a single membrane. Since the 20th week, microvilli were relatively scanty and poorly developed. Organelles and inclusions in the cytoplasm of metaplastic cells were poor. Nuclei were variable in shape. The nlost prominent changes at later stage were separation of nuclei from the cytoplasm, and appearance of numerous and irregularly angled electron dense granules in the nuclei.

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Neuronal Activity of the Vestibular Nuclei Following Acute Hypotension in Rats

  • Park, Byung-Rim;Kim, Min-Sun;Baik, Kum-Hyun;Lee, Moon-Young;Choi, Myung-Ae;Lee, Jae-Hyo
    • The Korean Journal of Physiology and Pharmacology
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    • v.6 no.4
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    • pp.199-205
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    • 2002
  • The role of peripheral vestibular receptors in acute hypotension was investigated in anesthetized rats. Acute hypotension was induced by either intravenous infusion of sodium nitroprusside (SNP) or by experimental hemorrhage, and electrical activity and expression of cFos-like immunoreactive (cFL) protein were measured in the medial vestibular nuclei (MVN). Blood pressure decreased proportionately to the does of intravenous SNP and to the volume of the hemorrhage. Blood pressure decreased 10, 30, 50% for the 5, 10, $15{\mu}g/kg$ SNP injection, respectively, and also decreased 30 and 50% after 1- and 2-ml blood loss, respectively, due to hemorrhage. In animals with intact labyrinths, acute hypotension induced by either intravenous infusion of SNP or hemorrhage produced different electrical activities with three different patterns in type I and II neurons of MVN. The responses of type I neurons showed excitatory in 2/3 of recorded neurons and inhibitory or no change in 1/3 of neurons, while the responses of type II neurons showed inhibitory in 2/3 of recorded neurons and excitatory or no change in 1/3 of neurons. In unilateral labyrinthectomized animals, 2/3 of type I neurons ipsilateral to the lesion showed an inhibitory response, and 2/3 of contralateral type I neurons showed an excitatory response after the induction of acute hypotension. The response patterns of type II neurons were opposite from those of the type I neurons. After 30% decrease in blood pressure, cFL protein expressed in the bilateral vestibular nuclei of control animals with intact labyrinths. Expression of cFL protein increased significantly proportionately to the reduction of blood pressure. The unilateral labyrinthectomized animals with acute hypotension produced expression of cFL neurons in contralateral vestibular nuclei to the lesion side, but not in ipsilateral vestibular nuclei. However, cFL protein was not expressed in bilateral vestibular nuclei after acute hypotension in bilateral labyrinthectomized animals. These results suggest that the peripheral vestibular receptors might play a significant role in controlling blood pressure following acute hypotension via activation of type I neurons and inhibition of type II neurons in the vestibular nuclei.

LINEAR POLARIZATION OF A DOUBLE PEAKED BROAD EMISSION LINE IN ACTIVE GALACTIC NUCLEI

  • Lee, Hee-Won
    • Journal of The Korean Astronomical Society
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    • v.44 no.2
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    • pp.59-65
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    • 2011
  • A small number of active galactic nuclei are known to exhibit prominent double peak emission profiles that are well-fitted by a relativistic accretion disk model. We develop a Monte Carlo code to compute the linear polarization of a double peaked broad emission line arising from Thomson scattering. A Keplerian accretion disk is adopted for the double peak emission line region and the geometry is assumed to be Schwarzschild. Far from the accretion disk where flat Minkowski geometry is appropriate, we place an azimuthally symmetric scattering region in the shape of a spherical shell sliced with ${\Delta}{\mu}=0.1$. Adopting a Monte Carlo method we generate line photons in the accretion disk in arbitrary directions in the local rest frame and follow the geodesic paths of the photons until they hit the scattering region. The profile of the polarized flux is mainly determined by the relative location of the scattering region with respect to the emission source. When the scattering region is in the polar direction, the degree of linear polarization also shows a double peak structure. Under favorable conditions we show that up to 0.6% linear polarization may be obtained. We conclude that spectropolarimetry can be a powerful probe to reveal much information regarding the accretion disk geometry of these active galactic nuclei.

Nucleus-phonon interactions of MCsSO4 (M = Na, K, or Rb) single crystals studied using spin-lattice relaxation time

  • Choi, Jae Hun;Kim, Nam Hee;Lim, Ae Ran
    • Journal of the Korean Magnetic Resonance Society
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    • v.18 no.1
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    • pp.15-23
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    • 2014
  • The structural properties and relaxation processes of $MCsSO_4$ (M = Na, K, or Rb) crystals were investigated by measuring the NMR spectra and spin-lattice relaxation rates $1/T_1$ of their $^{23}Na$, $^{39}K$, $^{87}Rb$, and $^{133}Cs$ nuclei. According to the NMR spectra, the $MCsSO_4$ crystals contain two crystallographically inequivalent sites each for the M and Cs ions. Further, the relaxation rates of all these nuclei do not change significantly over the investigated temperature range, indicating that no phase transitions occur in these crystals in this range. The variations in the $1/T_1$ values of the $^{23}Na$, $^{39}K$, $^{87}Rb$, and $^{133}Cs$ nuclei in these three crystals with increasing temperature are approximately proportional to $T^2$, indicating that Raman processes may be responsible for the relaxation. Therefore, for nuclear quadrupole relaxation of the $^{23}Na$, $^{39}K$, $^{87}Rb$, and $^{133}Cs$ nuclei, Raman processes with n = 2 are more effective than direct processes.

Nuclear Localization Signal of Human Foamy Virus Integrase (인간 포미바이러스 인테그라제의 핵위치 신호)

  • Oh Soo-A;Kang Seung-Yi;Han Sung-Tae;An Dog-Gn;Shin Cha-Gyun
    • YAKHAK HOEJI
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    • v.50 no.2
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    • pp.93-98
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    • 2006
  • Human foamy virus (HFV) integrase mediates integration of viral c-DNA into cellular DNA. In this process, HFV prointegration complex (PIC) in which integrase is a key component moves to nuclei of the infected cells and leads to integration of viral DNA to the cellular genome, which is essential in viral life cycle. In general nuclear localization signals (NLS) have been suggested to be involved in localizing retroviral PIC to nuclei, but the mechanisms for nuclear localization of the HFV PIC remains unclear. To functionally identify the NLS of HFV integrase, various subdomains of the protein were expressed as GFP fusions and their subcellular locations were analyzed with confocal laser scanning microscopy. Wild type HFV integrase was karyophilic by targeting the fusion protein to nuclei of the COS-1 and 293T cells. Our results showed that strong NLS of HFV integrase was mapped to the C-terminal regions. In addition the karyophilic properties of N-terminal and central regions are not individually strong enough to direct localization of the fusion proteins to nuclei, but their cooperative activity for nuclear import was confirmed.

Characterization of the Variability of Nucleoli in the Cells of Panax ginseng Meyer In Vivo and In Vitro

  • Khrolenko, Yuliya A.;Burundukova, Olga L.;Lauve, Lyudmila S.;Muzarok, Tamara I.;Makhan'kov, Vyacheslav V.;Zhuravlev, Yuri N.
    • Journal of Ginseng Research
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    • v.36 no.3
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    • pp.322-326
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    • 2012
  • Results of karyological study of intact plants and some callus lines of Panax ginseng are presented. In the native plants of P. ginseng the nucleus with 1 nucleolus (90%) dominate, and nucleus with 2 nucleoli is rare. One nucleolar nucleus also dominate in interphase nuclei of cells of cultivated P. ginseng (from 2006), but we also found nucleus with 2 to 3 nucleoli in the same cell lines. Interphase nuclei of P. ginseng in long cultivated lines (from 1988) contain 1 to 9 nucleoli, with a predominance of nuclei containing from 3 to 4 nucleoli. It was shown that long-time cells (cultivated since 1988) had cytogenetic changes such as increase level of polyploid and aneuploid cells, increase of nucleoli number into interphase nucleus and decrease of nuclei/nucleoli ratio. These long-time cultivated cells had very low ginsenoside content.

Capability of Accumulation Mode Aerosols Containing Black Carbon as CCN Observed during the PACDEX Campaign (PACDEX 캠페인 자료로 분석한 블랙카본을 포함한 축적모드 에어로솔의 구름응결핵 가능성)

  • Lee, Si-Hye;Ghim, Young-Sung;Kim, Sang-Woo;Yoon, Soon-Chang
    • Journal of Korean Society for Atmospheric Environment
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    • v.26 no.4
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    • pp.380-391
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    • 2010
  • Airborne in-situ measurements of aerosol/cloud number concentrations were analyzed to investigate the effects of aerosols on warm cloud formation in the Pacific Dust Experiment (PACDEX) during April and May 2007. In the air masses originating from the Asian continent, high concentrations of fine particles including black carbon (BC) were observed when compared to other regions. A strong correlation (r=0.88) between condensation nuclei (CN) having sizes ranging from 0.1 to 1.0 mm ($CN_{0.1-1.0}$) and cloud condensation nuclei (CCN) at 0.4% supersaturation ($CCN_{0.4%}$) suggests that most of the $CN_{0.1-1.0}$ can contribute to cloud formation. The possibility of a cloud droplet formation by BC particles was expected at the high water vapor mixing ratio (WVMR) and the abundance of water-soluble components at the low altitude less than 3 km.