• Title/Summary/Keyword: Molecular size

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Department of DNA Chromatographic System for On-Site Detection of Food-Contaminating Bacteria (식중독균 현장탐지를 위한 DNA 크로마토그래피 분석시스템의 개발)

  • 김석하;정우성;백세환
    • KSBB Journal
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    • v.18 no.3
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    • pp.190-196
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    • 2003
  • An analytical system detecting DNA particularly utilizing a concept of membrane strip chromatography initially applied to home-version tests for, such as, pregnancy and ovulation has been developed. We have chosen S. typhimurium as model analyte among food-contaminating microorganisms that occurred in high frequencies, and invA gene, as a detection target, specific to Salmonella species. This gene was able to be amplified by PCR under optimal conditions employing newly designed primers in our laboratory. The PCR product was specifically measured via hybridization between the analyte and a DNA probe, which was a totally different feature from the conventional gel electrophoresis detecting the products based only on the molecular size. It is notable thar the DNA probe sequence was specially designed such that no separation of excess primers present after PCR was required. This was immobilized on a nitrocellulose (NC) membrane via streptavidin-biotin linkage minimizing a steric effect when the hybridization with the amplified DNA took place. The analyrical system detected the microorganism in a concentration of minimum $10^3$ cfu/mL (i.e., 10 cells per system), estimated from the standard curve, 20 to 40 minutes after adding the sample. This sneitivity was approximately 10 times higher than that of gel electrophoresis as an analytical tool conventionally used. Furthermore, the assay was able to be run at room temperature, which would ofter an extra advantage to users.

Extrahypothalamic Expression of Rat Growth Hormone Releasing Hormone (GHRH);a possible intrapituitary factor for lactotroph differentiation? (흰쥐의 시상하부외 지역에서의 Growth Hormone Releasing Hormone (GHRH) 유전자발현;뇌하수체내 국부인자로서 Lactotroph분화에 관여할 가능성에 대하여)

  • Lee, Sung-Ho
    • Clinical and Experimental Reproductive Medicine
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    • v.23 no.3
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    • pp.269-275
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    • 1996
  • Biosynthesis and secretion of anterior pituitary hormones are under the control of specific hypothalamic stimulatory and inhibitory factors. Among them, Growth Hormone Releasing Hormone (GHRH) is the major stimulator of pituitary somatotrophs activating GH gene expression and secretion. Human GHRH is a polypeptide of 44 amino acids initially isolated from pancreatic tumors, and the gene for the hypothalamic form of GHRH is organized into 5 exons spanning over 10 kilobases (kb) on genomic DNA and encodes a messenger RNA of 700-750 nucleotides. Several neuropeptides classically associated with the hypothalamus have been found in the extrahypothalamic regions, suggesting the existence of novel sources, targets and functions. GHRH-like immunoreactivity has been found in several peripheral sites, including placenta, testis, and ovary, indicating that GHRH may also have regulatory roles in peripheral reproductive organs. Furthermore, higher molecular weight forms of the GHRH transcripts were identified from these organs (1.75 kb in testis; 1.75 and >3 kb in ovary). These tissue-specific expression of GHRH gene suggest the existence of unique regulatory mechanism of GHRH expression and function in these organs. In fact, placenta-specific and testis-specific promoters for GHRH transcripts which are located in about 10 kb upstream region of hypothalamic promoter were reported. The use of unique promoters in extrahypothalamic sites could be refered in a different control of GHRH gene and different functions of the translated products in these tissues. Somatotrophs and lactotrophs have been thought to be derived from a common bipotential progenitor, the somatolactotrophs, which give origins to either phenotypes. Although the precise mechanism responsible for the lactotroph differentiation in the anterior pituitary gland has not been yet clalified, there are several candidators for the generation of lactotrophs. In human, the presence of GHRH peptides with different size from authentic hypothalamic form in the normal anterior pituitary and several types of adenoma were demonstrated. Recently our group found the existence of immunoreactive GHRH and its transcript from the normal rat anterior pituitary (gonadotroph> somatotroph> lactotroph), and the GHRH treatment evoked the increased proliferation rate of anterior pituitary cells in vitro. The transgenic mouse models clearly shown that GHRH or NGF overexpression by anterior pituitary cells induced development of pituitary hyperplasia and adenomas particularly GH-oma and prolactinoma. Taken together, we hypothesize that the pituitary GHRH could serve not only as a modulator of hormone secretion but as a paracrine or autocrine regulator of anterior pituitary cell proliferation and differentiation. Interestingly enough, the expression of Pit-1 homeobox gene (the POU class transcription factor) was confined to somatotrophs, lactotrophs and somatolactotrophs in which GHRH receptors are expressed commonly. Concerning the mechanism of somatolactotroph and lactotroph differentiation in the anterior pituitary, we have focused following two possibilities; (1) changes in the relative levels or interactions of both hypothalamic and intrapituitary factors such as dopamine, VIP, somatostatin, NGF and GHRH; (2) alterations of GHRH-GHRH receptor signaling and Pit-1 activity may be the cause of lactotroph differentiation or pituitary hyperplasia and adenoma formation. Extensive further studies will be necessary to solve these complicated questions.

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Local Drug Delivery System Using Biodegradable Polymers

  • Khang, Gil-Son;Rhee, John M.;Jeong, Je-Kyo;Lee, Jeong-Sik;Kim, Moon-Suk;Cho, Sun-Hang;Lee, Hai-Bang
    • Macromolecular Research
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    • v.11 no.4
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    • pp.207-223
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    • 2003
  • For last five years, we are developing the novel local drug delivery devices using biodegradable polymers, especially polylactide (PLA) and poly(D,L-lactide-co-glycolide) (PLGA) due to its relatively good biocompatibility, easily controlled biodegradability, good processability and only FDA approved synthetic degradable polymers. The relationship between various kinds of drug [water soluble small molecule drugs: gentamicin sulfate (GS), fentanyl citrate (FC), BCNU, azidothymidine (AZT), pamidronate (ADP), $1,25(OH)_2$ vitamin $D_3$, water insoluble small molecule drugs: fentanyl, ipriflavone (IP) and nifedipine, and water soluble large peptide molecule drug: nerve growth factor (NGF), and Japanese encephalitis virus (JEV)], different types of geometrical devices [microspheres (MSs), microcapsule, nanoparticle, wafers, pellet, beads, multiple-layered beads, implants, fiber, scaffolds, and films], and pharmacological activity are proposed and discussed for the application of pharmaceutics and tissue engineering. Also, local drug delivery devices proposed in this work are introduced in view of preparation method, drug release behavior, biocompatibility, pharmacological effect, and animal studies. In conclusion, we can control the drug release profiles varying with the preparation, formulation and geometrical parameters. Moreover, any types of drug were successfully applicable to achieve linear sustained release from short period ($1{\sim}3$ days) to long period (over 2 months). It is very important to design a suitable formulation for the wanting period of bioactive molecules loaded in biodegradable polymers for the local delivery of drug. The drug release is affected by many factors such as hydrophilicity of drug, electric charge of drug, drug loading amount, polymer molecular weight, the monomer composition, the size of implants, the applied fabrication techniques, and so on. It is well known that the commercialization of new drug needs a lot of cost of money (average: over 10 million US dollar per one drug) and time (average: above 9 years) whereas the development of DDS and high effective generic drug might be need relatively low investment with a short time period. Also, one core technology of DDS can be applicable to many drugs for the market needs. From these reasons, the DDS research on potent generic drugs might be suitable for less risk and high return.

An Ultrafiltration Study for the Recycling of Synthetic Water-Based Cutting Oil (수용성 합성 절삭유의 재사용을 위한 한외여과 연구)

  • Kim, Jong-Pyo;Kim, Jae-Jin;Ryu, Jong-Hoon
    • Clean Technology
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    • v.8 no.3
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    • pp.119-128
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    • 2002
  • In the present study the membrane filtration characteristics of a commercially available synthetic water-based cutting oil through two kinds of ultrafiltration membranes (HF1-45-CM50 and HF1-43-CM100) with molecular weight cut-offs of 50,000 and 100,000, respectively, have been investigated in detail. Among these membranes, the hydrophilic one (HF1-45-CM50) was found to show a satisfactory result for both the permeate flux and the permeability of oil components, whereas the permeate flux obtained with the hydrophobic membrane (HF1-43-CM100) appears to be significantly low, indicating that synthetic cutting oil was easily wetted on the hydrophobic membrane surface and induced more membrane fouling. The effect of material characteristics of the membrane on the filtration characteristics was found to be much more significant compared with the mean pore size of the membrane. Backflushing by nitrogen gas was applied to reduce the formation of a gel layer and membrane fouling. With the hydrophilic membrane, the backflushing was found to increase the permeate flux, whereas the backflushing resulted in a decrease in flux for the hydrophobic membrane. The flux recovery was observed to be highest when the membranes fouled with waste synthetic cutting oil were immersed into a cleaning solution for more than 72 hours and then backflushed by nitrogen gas.

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MicroRNA-766-3p Inhibits Tumour Progression by Targeting Wnt3a in Hepatocellular Carcinoma

  • You, Yu;Que, Keting;Zhou, Yun;Zhang, Zhen;Zhao, Xiaoping;Gong, Jianpin;Liu, Zuojin
    • Molecules and Cells
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    • v.41 no.9
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    • pp.830-841
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    • 2018
  • Recent studies have indicated that microRNAs (miRNAs) play an important role in hepatocellular carcinoma (HCC) progression. In this study, we showed that miR-766-3p was decreased in approximately 72% of HCC tissues and cell lines, and its low expression level was significantly correlated with tumour size, TNM stage, metastasis, and poor prognosis in HCC. Ectopic miR-766-3p expression inhibited HCC cell proliferation, colony formation, migration and invasion. In addition, we showed that miR-766-3p repressed Wnt3a expression. A luciferase reporter assay revealed that Wnt3a was a direct target of miR-766-3p, and an inverse correlation between miR-766-3p and Wnt3a expression was observed. Moreover, Wnt3a up-regulation reversed the effects of miR766-3p on HCC progression. In addition, our study showed that miR-766-3p up-regulation decreased the nuclear ${\beta}-catenin$ level and expression of Wnt targets (TCF1 and Survivin) and reduced the level of MAP protein regulator of cytokinesis 1 (PRC1). However, these effects of miR-766-3p were reversed by Wnt3a up-regulation. In addition, PRC1 upregulation increased the nuclear ${\beta}-catenin$ level and protein expression of TCF1 and Survivin. iCRT3, which disrupts the ${\beta}-catenin-TCF4$ interaction, repressed the TCF1, Survivin and PRC1 protein levels. Taken together, our results suggest that miR-766-3p down-regulation promotes HCC cell progression, probably by targeting the Wnt3a/PRC1 pathway, and miR-766-3p may serve as a potential therapeutic target in HCC.

Condensable Gas Separation using Phenol! Alumina Composite Activated Carbon Hollow Fiber Membranes (페놀수지/알루미나 복합 활성탄소중공사막을 이용한 응축성 기체 분리)

  • Shin, Kyung-Yong;Park, You-In;Kim, Beom-Sik;Koo, Kee-Kahb
    • Membrane Journal
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    • v.20 no.4
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    • pp.312-319
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    • 2010
  • Carbon membrane materials have received considerable attention for the gas separation including hydrocarbon mixture of ingredients of the volatile organic compounds(VOCs) because they possess their higher selectivity, permeability, and thermal stability than the polymeric membranes. The use of activated carbon membranes makes it possible to separate continuously the VOCs mixture by the selective adsorption-diffusion mechanism which the condensable components are preferentially adsorbed in to the micropores of the membrane. The activated carbon hollow fiber membranes with uniform adsorptive micropores on the wall of open pores and the surface of the membranes have been fabricated by the carbonization of a thin film of phenolic resin deposited on porous alumina hollow fiber membrane. Oxidation, carbonization, and activation processing variables were controlled under different conditions in order to improve the separation characteristics of the activated carbon membrane. Properties of activated carbon hollow fiber membranes and the characterization of a gas permeation by pyrolysis conditions were studied. As the result, the activated carbon hollow fiber membranes with good separation capabilities by the molecular size mechanism as well as selective adsorption on the pores surface followed by surface diffusion effective in the recovery hydrocarbons have been obtained. Therefore, these activated carbon membranes prepared in this study are shown as promising candidate membrane for separation of VOCs.

Recent Progress on Metal-Organic Framework Membranes for Gas Separations: Conventional Synthesis vs. Microwave-Assisted Synthesis (기체분리용 금속유기구조체 분리막의 최근 연구 동향 및 성과)

  • Ramu, Gokulakrishnan;Jeong, Hae-Kwon
    • Membrane Journal
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    • v.27 no.1
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    • pp.1-42
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    • 2017
  • Metal-organic frameworks (MOFs) are nanoporous materials that consist of organic and inorganic moieties, with well-defined crystalline lattices and pore structures. With a judicious choice of organic linkers present in the MOFs with different sizes and chemical groups, MOFs exhibit a wide variety of pore sizes and chemical/physical properties. This makes MOFs extremely attractive as novel membrane materials for gas separation applications. However, the synthesis of high-quality MOF thin films and membranes is quite challenging due to difficulties in controlling the heterogeneous nucleation/growth and achieving strong attachment of films on porous supports. Microwave-based synthesis technology has made tremendous progress in the last two decades and has been utilized to overcome some of these challenges associated with MOF membrane fabrication. The advantages of microwaves as opposed to conventional synthesis techniques for MOFs include shorter synthesis times, ability to achieve unique and complex structures and crystal size reductions. Here, we review the recent progress on the synthesis of MOF thin films and membranes with an emphasis on how microwaves have been utilized in the synthesis, improved properties achieved and gas separation performance of these films and membranes.

A refined Panax ginseng karyotype based on an ultra-high copy 167-bp tandem repeat and ribosomal DNAs

  • Waminal, Nomar Espinosa;Choi, Hong-Il;Kim, Nam-Hoon;Jang, Woojong;Lee, Junki;Park, Jee Young;Kim, Hyun Hee;Yang, Tae-Jin
    • 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.

The Effect of Surfactant on Controlled Release of Amino acids Through Poly(2-Hydroxyethyl Methacrylate) Membrane (Poly(2-Hydroxyethyl Methacrylate)막을 통한 아미노산의 방출 조절에 대한 계면활성제의 효과)

  • Kim Ui-Rak;Jeong Bong-Jin;Lee Myung-Jae;Min Kyung-Sub
    • Journal of the Korean Chemical Society
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    • v.37 no.1
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    • pp.22-35
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    • 1993
  • The transport phenomena of the free amino acids through poly(hydroxyethyl methacrylate)[P(HEMA)] have been investigated with and without various kinds of surfactants solution and in the mixed surfactants solution. Glutamine has the highest diffusivity among 4 amino acids at 1CMC of cetyldimethylethylammonium bromide(CTABr) surfactant. Glutamic acid is not affected by the concentration of CTABr. Methionine and Lysine shows slight decreased diffusivity at 0.5 CMC, but increase its diffusivity at 1CMC and 2CMC due to the structure change of membrane and the viscosity change of surfactant solution. Glutamic acid has the highest diffusivity among four amino acids at sodium dodecyl sulfate(SDS) and Triton X-100 surfactant. In mixed surfactant solution, each amino acids shows high diffusivity through 45% water content membrane at the 0.5 mole fraction of SDS in the SDS/TX-100 surfactant mixtures. It has been found that not only the property of membrane but also the effects of solute-solvent interactions and solvent effect are very important as the permeation of amino acids occurs through P(HEMA) membrane. The diffusivities of free amino acids through membrane depend upon their molecular shape, size and charge.

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Expression of mRNAs characteristic of cartilage and bone in the developing mandibular condyle of mice (발육중인 생쥐 하악 과두에서 연골 및 골의 특이 유전자 발현)

  • Ji, Kuk-Soep;Yoon, Young-Jooh;Park, Joo-Cheol;Kim, Kwang-Won
    • The korean journal of orthodontics
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    • v.34 no.2 s.103
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    • pp.143-152
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    • 2004
  • It has not been elucidated whether the initiation of condylar development of the mandible is related with the periosteum of the mandible, or if it derives from a separate programmed blastema not related with the mandible. Also, although the mandibular condylar cartilage is known to promote growth, few studies have dealt with molecular-biologic mechanisms such as the expression of specific genes according to the differentiation of the mandibular condyle. To elucidate the unique cellular characteristics, development, and differentiation process of the mandibular condyle, an examination of expressions of genes characteristic of cartilage and bone were carried out using RT-PCR and mRNA in situ hybridization. 1. Type? collagen mRNA was detected with type II collagen mRNA in the differentiation and growth process of the cartilage of the mandibular condyle. TypeII collagen mRNA was demonstrated in the whole resting md upper part of the poliferative zone, whereas type II collagen mRNA was observed in the resting, proliferative and upper hypertrophic cartilage zone of the mandibular condyle. 2. The condylar cartilage rapidly increased in size due to the accumulation of hypertrophic chondrocytes as characterized by the expression of type II collagen mRNA during postnatal development. 3. BMP-4 mRNA was present in the anlage of the future condylar process and also in the ossifying mandibular body. 4. IHH mRNA was limited exclusively to the lower part of the proliferative zone and the upper part of the hypertrophic cartilage zone during condylar development. These findings were different from those in the growth-plate cartilage of the long bone, indicating a characteristic feature of the differentiation of the chondrocytes in the condylar cartilage present in prenatal and postnatal development. Furthermore, it was also suggested that chondroblasts of condylar cartilage rapidly differentiate into hypertrophic chondrocytes with increased functional Load force such as muscle activity and mastication.