• Journal of Applied Biological Chemistry
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• v.58 no.3
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• pp.273-279
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• 2015

Phaeodactylum tricornutum is a model diatom that its genomic information and biological tools are well established. In this study, a gene encoding (E)-4-hydroxy-3-methylbut-2-enyl diphosphate reductase (PtHDR), a terminal enzyme of the methylerythritol phosphate pathway regulating chlorophyll and carotenoid biosynthesis, was isolated from P. tricornutum. The isolated gene was cloned into pPha-T1 vector containing fcpA promoter to prepare pPha-T1-HDR plasmid. As a positive control, pPha-T1-eGFP plasmid was constructed with egfp gene. Stable nuclear transformation was carried out with these plasmids by particle bombardment method and zeocin resistant colonies of P. tricornutum were selected on f/2 agar plate. In result, transformation efficiency was evaluated according to the amount of plasmid DNA coated with gold particles. Integration of introduced plasmids was confirmed with genomic DNA of each transformant by polymerase chain reaction. The eGFP fluorescence was visible in the cytoplasm, indicating that eGFP was successively expressed in P. tricornutum system. The transcript level of exogenous Pthdr gene was evaluated with the obtained transformants. The results presented here demonstrated that introduction of Pthdr gene into P. tricornutum chromosome succeeded and expression of PtHDR was enhanced under the fcpA promoter.

• Applied Microscopy
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• v.15 no.1
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• pp.31-58
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• 1985

The morphological study on different types of cells of reproductive organ including spermatogenesis in the adult planaria was performed to observe their cytochemical and ultrastructural characteristics. 1. Spermatogenesis The circular luminated material appears immediately inside the nuclear envelope of early spermatid and is found also in the nucleus of sperm, but typical acrosomal structures cannot be observed. Approximately ten of small-sized mitochondria occur around the nucleus in the transitional phase from primary spermatocyte to secondary spermatocyte, but in sperm a long mitochondrion is closely associated with nucleus, parellel to long axis of it. The sperm has a relatively long head connected with two tails via hollow neck. 2. Reproductive organ The penis bulb and the bursa stalk were observed. (1) Penis bulb The cells constituted penis bulb are classified into six types on the basis of ultrastructure of the cells and cytochemistry of the cytoplasmic granules. 1) A-type cells: These cells exhibiting low electron density are mainly occupied by large nucleus. These cells possess two different types of granules: highly electron-dense round granules with an average size of $0.9{\mu}m$, and electron-dense granules exhibit PAS-positive reaction. 2) B-type cells contain PAS-positive granules with the size of about $0.4{\mu}m$. They are rich in free ribosomes and mitochondria. 3) C-type cells are found to be dark cells due to high electron-density. These cells are largely occupied by large nucleus. 4) D-type cells: These cells are seen as light cells which have poorly developed cell organelles. 5) E-type tells: These cells contain a large number of glycogen granules which occupy most of cell. 6) F-type cells: These arc parietal epidermal cells surrounding the genital antrum. These cells are characterized by their finger-like shapes and the presence of a number of electron-dense, irregularly-shaped structures inside cells. The relatively large electron-lucent granules can be also found. The F-type cells possess numerous microvilli on their free surfaces. (2) Bursa stalk The cells constituted bursa stalk are classified into 3 types on the basis of cell shapes and presences of electron-dense or electron-lucent granules. 7) G-type cells with a long cytoplasmic process. They have large nuclei and poorly developed cell organelles. 8) H-type cells: These cells are characterized by the presence of a long cytoplasmic process and relatively highly electron-dense cytoplasmic profile. They have poorly developed cell organelles. 9) I-type cells contain large electron-lucent granules which exhibit negative reactions with three kinds of cytochemical staining methods used in this experiment. The fine electron-dense structures can be found inside these granules.

• Journal of Embryo Transfer
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• v.25 no.2
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• pp.127-131
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• 2010

The objective of this study was to determine the mRNA expression patterns of several putative imprinted genes in in vivo and in vitro fertilized, parthenogenetic, and cloned porcine preimplantation embryos. Both maternally (Dlk1, IGF2, Peg1/Mest and Ndn) and paternally (IGF2r, H19 and Xist) imprinted genes were selected. We have used reverse transcription polymerase chain reaction (RT-PCR) to investigate gene expression patterns in the porcine embryos. IGF2 transcripts were detected in the most of embryos. In nuclear transfer (NT), Peg1/MEST transcripts showed fluctuating pattern. Dlk1 was only expressed partially from the morula and blastocyst stage of NT embryos. Ndn gene expression was started somewhat early for in vivo embryos. However, the expressions of maternally imprinted genes were similar in all types of blastocysts (NT, in vivo and in vitro fertilized, and parthenogenetic embryos). The IGF2R gene expression level was somewhat irregular and varied among samples. However, for the majority samples of all types of embryos, IGF2R expression was diminished after one- to two-cell stages and reappeared at the morulae or blastocyst stage embryos. H19 gene was only expressed early in parthenogenetic and in vivo embryos. For NT embryos, H19 was only expressed in blastocysts. Xist expression was detected in all blastocysts with the earliest being in vivo 8-cell stage embryos and the last one being NT blastocysts. These putative imprinted genes appeared to have stage specific expression patterns with a fluctuating pattern for some genes (Peg/Mest, IGF2r, H19). These results suggest that stage specific presence of imprinted genes can affect the embryo implantation and fetal development.

• Journal of Life Science
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• v.15 no.3 s.70
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• pp.323-331
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• 2005

The objective of the present study was to investigate the effect of $\beta-lapachone$, a quinone obtained from the bark of the lapacho tree (Tabebuia avellanedae) in South America, on the cell growth of human hepatoma (HepG2) and bladder (T24) carcinoma cells. Exposure of cancer cells to $\beta-lapachone$ resulted in growth inhibition, morphological changes and apoptosis in a concentration-dependent manner, which could be proved by MTT assay and flow cytometry analysis. Reverse transcription-polymerase chain reaction (RT-PCR) and Western blot analyses revealed that $\beta-lapachone$ did not affect the levels of tumor suppressor p53 and cyclin-dependent kinase (Cdk) inhibitor p21 (WAFl/CIPl) expression. However, the transcriptional factor Sp-l and proliferating cell nuclear antigen (PCNA) protein levels were significantly down-regulated by $\beta-lapachone$ in both cell lines. Moreover, $\beta-lapachone$ treatment caused a dose-dependent inhibition of the expression of telomere regulatory gene products such as human telomere reverse transcriptase (hTERT) and telomerase-associated protein-l (TEP-l). Taken together, these findings suggest that $\beta-lapachone$-induced inhibition of human hepatoma and bladder carcinoma cell proliferation is associated with the induction of apoptotic cell death via modulation of several major growth regulatory gene products, and provide important new insights into the additional mechanisms of the anti-cancer activity of $\beta-lapachone$.

• Applied Chemistry for Engineering
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• v.17 no.5
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• pp.552-556
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• 2006

An advanced oxidation process catalyzed by iron ions in the presence of hydrogen peroxide, the so-called Fenton's reaction, has been applied to the treatment of steam generator chemical cleaning waste containing highly concentrated iron(III)- ethyl-enediaminetetraaceticacid (Fe(III)-EDTA) of 70000 mg/L. The experiments for the degradation of Fe(III)-EDTA were carried out not only with a simulated waste, but also with the real one. The effect of pH and the amount of hydrogen peroxide added to the waste on the degradation was examined, and the results were discussed in several aspects. The optimal pH to maximize the degradation efficiency was dependent on the amount of hydrogen peroxide added to the waste. i.e., when the amount of hydrogen peroxide was different, maximum degradation efficiency was obtained at different pH's. The optimal amount of hydrogen peroxide relative to that of Fe(III)-EDTA was found to be 24.7 mol ($H_{2}O_{2}$)/mol (Fe(III)-EDTA) at pH around 9.

• Journal of Life Science
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• v.14 no.2
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• pp.215-220
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• 2004

We investigated the anti-inflammatory effects of aqueous extract of Artemisia capillaris Thunb. (AEAC), a traditional Korean herb for remedying liver disease, for suppression in the process of lipopolysaccharide (LPS)-induced inflammation in the liver of rat. Level of glutamate oxaloacetate transaminase (GOT), glutamate pyruvate transaminase (GPT) and lactate dehydrogenase (LDH) was increased in the serum of LPS-treated rats compared to normal, however, in the rats pretreated with AEAC, the increase of GOT, GPT and LDH value was arrested. More severe histological changes of liver such as cloudy swelling, hydropic degeneration, Kupffer cell reaction and inflammatory cells infiltration were demonstrated in the rats challenged with LPS compared with normal. Fewer scores of these changes were observed in rats pretreated with AEAC. Immunohistochemical analysis showed that while the expression of the nuclear factor (NF)-kBp65, inducible nitric oxide synthase (iNOS), tumor necrosis factor (TNF)-$\alpha$ and COX (cyclooxygenase)-2 tended to increase, that of inhibitory (I)-kBa was decreased in the hepatocytes of rats challenged with LPS. A slight decline of NF-kBp65, TNF-$\alpha$ and COX-2, but increase of I-kB$\alpha$ were observed in the hepatocytes of the rats pretreated with AEAC. These results suggest that AEAC may act as a therapeutic agent for liver disease through a regulation of inflammation-related proteins.

• Journal of Radiation Protection and Research
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• v.20 no.2
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• pp.79-83
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• 1995

A number of investigators have reported formation of radiolytic ultrafine particles produced by the interaction of ionizing radiation with atmospheric trace gases. Previous studies have suggested that a very high localized concentration of the hydroxyl radical produced by the radiolysis of water can react with atmospheric trace gases such as $SO_2$ and produce lower vapor pressure compounds that can subsequently nucleate. To determine the trace gas and water vapor concentration dependence of the active, positively charged, first decayt product of radon (Po-218), a well-controlled radon chamber was used in this research. The mobility spectrum of the decay products in the range of $0.07-5.0cm^2/V\;sec$ from the radon chamber was measured using alpha track detector installed inside a specially-designed electrostatic spectrometer. Measurements were taken for different concentrations (0.5ppm to 5ppm) of $SO_2$ in Purified, Compressed air. A kinetics Study following the clustering of $SO_2$ around the $PoO_x^+$ ion in an excess of $SO_2$ for interpretation of the reaction processes was performed.

• Korean Journal of Materials Research
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• v.8 no.12
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• pp.1099-1109
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• 1998

To investigate the effect of V and Sb on the corrosion behavior of Zr- based alloys, corrosion tests were performed on 6 kinds of Zr alloys in an autoclave at $360^{\circ}C$ for 100 days. The transition of the corrosion rate occurred in the sample containing 0.1wt.%V after 10 days but did not occur in the samples containing 0.2wt.%V and 0.4wt.%V. The corrosion resistance of V containing alloys increased with increasing V contents from 0.1 to 0.4wt.% and the alloys containing 0.4wt.%V showed the best corrosion resistance. In the ternary alloys containing 0.1wt.%Sb and 0.4wt.%Sb, the corrosion rate increased significantly from the short exposure time. It was observed that the optimal Sb content for corrosion resistance was 0.2wt.%. The size and volume fraction of precipitates increased with increasing V and Sb contents. The superior corrosion resistance was observed in the Zr alloy having precipitate size of 0.11-0.13$\mu\textrm{m}$. From the result of corrosion behavior and the obserbation of precipitates, the optimal size of the precipitate appear to control the electron conduction in the cathodic reaction and play an important role in maintaining a stable oxide microstructure.

• Journal of Ginseng Research
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• v.41 no.4
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• pp.469-476
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• 2017

Background: Panax ginseng Meyer (Asian ginseng) has a large nuclear genome size of > 3.5 Gbp in haploid genome equivalent of 24 chromosomes. Tandem repeats (TRs) occupy significant portions of the genome in many plants and are often found in specific genomic loci, making them a valuable molecular cytogenetic tool in discriminating chromosomes. In an effort to understand the P. ginseng genome structure, we characterized an ultrahigh copy 167-bp TR (Pg167TR) and explored its chromosomal distribution as well as its utility for chromosome identification. Methods: Polymerase chain reaction amplicons of Pg167TR were labeled, along with 5S and 45S rDNA amplicons, using a direct nick-translation method. Direct fluorescence in situ hybridization (FISH) was used to analyze the chromosomal distribution of Pg167TR. Results: Recently, we reported a method of karyotyping the 24 chromosome pairs of P. ginseng using rDNA and DAPI (4',6-diamidino-2-phenylindole) bands. Here, a unique distribution of Pg167TR in all 24 P. ginseng chromosomes was observed, allowing easy identification of individual homologous chromosomes. Additionally, direct labeling of 5S and 45S rDNA probes allowed the identification of two additional 5S rDNA loci not previously reported, enabling the refinement of the P. ginseng karyotype. Conclusion: Identification of individual P. ginseng chromosomes was achieved using Pg167TR-FISH. Chromosome identification is important in understanding the P. ginseng genome structure, and our method will be useful for future integration of genetic linkage maps and genome scaffold anchoring. Additionally, it is a good tool for comparative studies with related species in efforts to understand the evolution of P. ginseng.

• Journal of Periodontal and Implant Science
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• v.42 no.6
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• pp.185-195
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• 2012

Purpose: (-)-epigallocatechin-3-gallate (EGCG) has been reported to exert anti-inflammatory and antibacterial effects in periodontitis. However, its exact mechanism of action has yet to be determined. The present in vitro study evaluated the anti-in-flammatory effects of EGCG on human periodontal ligament fibroblasts (hPDLFs) and human periodontal ligament stem cells (hPDLSCs) affected by bacterial lipopolysaccharide (LPS) extracted from Porphyromonas gingivalis. Methods: hPDLFs and hPDLSCs were extracted from healthy young adults and were treated with EGCG and/or P. gingivalis LPS. After 1, 3, 5, and 7 days from treatment, cytotoxic and proliferative effects were evaluated using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and bromodeoxyuridine assay, respectively. And then, the gene expressions of hPDLFs and hPDLSCs were observed for interleukin (IL)-$1{\beta}$, IL-6, tumor necrosis factor (TNF)-${\alpha}$, osteoprotegerin (OPG), receptor activator of nuclear factor kappa-B ligand (RANKL), and RANKL/OPG using real-time polymerase chain reaction (PCR) at 0, 6, 24, and 48 hours after treatment. The experiments were performed with the following groups for hPDLFs and hPDLSCs; 1) No treat, 2) EGCG alone, 3) P. gingivalis LPS alone, 4) EGCG+P. gingivalis LPS. Results: The 20 ${\mu}M$ of EGCG and 20 ${\mu}g/mL$ of P. gingivalis LPS had the lowest cytotoxic effects, so those concentrations were used for further experiments. The proliferations of hPDLFs and hPDLSCs increased in all groups, though the 'EGCG alone' showed less increase. In real-time PCR, the hPDLFs and hPDLSCs of 'EGCG alone' showed similar gene expressions to those cells of 'no treat'. The gene expressions of 'P. gingivalis LPS alone' in both hPDLFs and hPDLSCs were highly increased at 6 hours for IL-$1{\beta}$, IL-6, TNF-${\alpha}$, RANKL, and RANKL/OPG, except the RANKL/OPG in hPDLSCs. However, those increased gene expressions were down-regulated in 'EGCG+P. gingivalis LPS' by the additional treatment of EGCG. Conclusions: Our results demonstrate that EGCG could exert an anti-inflammatory effect in hPDLFs and hPDLSCs against a major pathogen of periodontitis, P. gingivalis LPS.