• The Journal of Korea Institute of Information, Electronics, and Communication Technology
• /
• v.15 no.5
• /
• pp.387-395
• /
• 2022

PDRN (Polydeoxyribonucleotide) is a DNA-derived polymer that promotes self-renewal of damaged cells and tissues as a tissue regeneration active material. PDRN is a DNA fragment cut into small sizes by various physical or chemical methods. When administered to the body, PDRN binds and stimulates the adenosine A2A receptor on the surface of tissue cells to promote cell regeneration, accelerate wound healing, and reduce pain. Although PDRN is prepared from testis or semen of fish in most cass, PDRN extraction from various plants species was performed in the present study. Among 7 tested plant species, the highest DNA yield and purity was obtained form mugwort (Chrysanthemum coronarium, C.c), followed by broccoli (Brassica oleracea, B.o). Then, we evaluated the in vitro wound healing capacity of PDRNs prepared from these two selected plants. PDRN from C.c and B.o. significantly stimulated the wound healing process at ㎍/ml range. The present study suggests that PDRN from plant species can be an effective alternative to PDRN from marine organism.

• Korean Journal of Oriental Medicine
• /
• v.12 no.3 s.18
• /
• pp.79-90
• /
• 2006

This study was investigated to determine the quality monitoring of distributed materials from Glycyrrhizae Radix (26 samples), Atractylodis Rhizoma Alba (24 samples) according to storage period after $1{\sim}3$ year. We have estimated by identification, purity, loss on drying, ash, acid insoluble ash, extract content, essential oil content, assay and microbial contamination. As a result, Glycyrrhizae Radix (26 samples) were satisfied with the standard of K.P. (Korean Pharmacopoeia) and WHO's microbial contamination limit standard. In the Atractylodis Rhizoma Alba (24 samples), 2 samples were not satisfied with the standard of K.P.(Korean Pharmacopoeia) and WHO's microbial contamination limit standard. The results make practical application of the basic data for the quality control of herbal medicine in storage.

• Korean Journal of Veterinary Service
• /
• v.28 no.4
• /
• pp.393-405
• /
• 2005

In prion pathogenesis, the structural conversion of the cellular prion protein $(PrP^c)$ to its abnormal isomer $(PrP^{Sc})$ is believed to be a major event. The susceptibility or resistance to natural sheep scrapie is associated with polymorphisms of host PrP gene (PRNP) at amino acid residues 136, to a lesser extent 154. The 112 residue in ovine PrP displays a natural polymorphism, Methionine to Threonine, which has not been thoroughly investigated. However the cell-free conversion assay showed that ARQ with Thr112 $(T_{112}ARQ)^{1)}$ presents lower convertibility to $PrP^{Sc}$than wild type ARQ $(M_{112}ARQ)$ [1] In this study we generated ovine recombinant PrPs of 112 allelic variants by metal chelate affinity chromatography and cation exchange chromatography. The final purity of the ovine PrP ARQ was more than $95\%$. These variants showed similar immunoreactivity against anti-PrP monoclonal antibodies in Western blot and ELISA. The refolded $M_{112}ARQ$ and $M_{112}ARQ$ presented the secondary structural content to similar extent via CD spectroscopy analysis. The inherited structural features of $M_{112}ARQ$ and $M_{112}ARQ$ under the different biophysical conditions are in the middle of investigation.

• The Korean Journal of Nuclear Medicine
• /
• v.38 no.6
• /
• pp.532-539
• /
• 2004

Purpose: There has been a renewal of interest in Macrophage migration inhibitory factor (MIF), especially correlation in pathogenesis of sepsis by many infectious diseases and in regulation of host inflammatory and immune response. We developed immunoradiometric assay (IRMA) to determine serum human MIF concentration. Materials and Methods: The IRMA system utilizes solid phase bound monoclonal anti-recombinant human MIF (rhMIF) antibody as a capture antibody, biotinylated polyclonal anti-rhMIF antibody as a detector antibody. We applied with rhMIF that concentration of standard solutions increased from 0 ng/ml to 100 ng/ml. We used $^{125}I$-streptavidin (SA) as radiotracer to determination of rhMIF concentration. Streptavidin was labeled with $^{125}I$ by Chloramine-T method and $^{125}I$-SA was purified by ultracentrifugation. $^{125}I$-SA stability was evaluated by ITLC analysis at $4^{\circ}C$ and room temperatures until 60days. To validate IRMA system for MIF, we experimented intra-assay and inter-assay coefficients of variation, recovery test and dilution test. Results: Radiolabeling yield of $^{125}I$-SA was 87% and purified $^{125}I$-SA retained above 99% radiochemical purity. $^{125}I$-SA showed above 93% stability in $4^{\circ}C$ until 60days that it is good for immunoradiometric assay as radiotracer. Plotted standard dose response curve showed that increased concentration of rhMIF linearly correlated (R2=0.99) with bound radioactivity of $^{125}I$-SA. The highest intra- and inter-assay coefficients of variation were 5.5% and 7.6%, respectively. The average of recovery of MIF in samples was 102%. In dilution test, linear response curves were obtained (R2=0.97). Conclusion: Radioimmunoassay using $^{125}I$-SA as radiotracer thought to be useful for the determination of serum MIF concentration, and further, its data will be used to evaluate the correlation between clinical significance and serum MIF concentration in patients with various inflammatory diseases.

• Korean Journal of Microbiology
• /
• v.41 no.3
• /
• pp.216-224
• /
• 2005

Chromatography has now been used successfully to provide the requisite purity for human plasma-derived biop-harmaceuticals such as coagulation factors and immunoglobulins. Recently, increasing attention has been focused on establishing efficient cleaning procedures to prevent potential contamination by microorganisms as well as carry-over contamination from batch to batch. The purpose of present study was to develop a cleaning validation system for the assurance of virus removal and/or inactivation during chromatography process. In order to establish an assay system for the validation of virus clearance during chromatography cleaning process, a quantitative real-time PCR method for porcine parvovirus(PPV) was developed, since PPV, a model virus for human parvovirus B19, has a high resistance to a range of physico-chemical treatment. Specific primers for amplification of PPV DNA was selected, and PPV DNA was quantified by use of SYBR Green I. The sensitivity of the assay was calculated to be 1.5 $TCID_{50}/ml$. The established real-time PCR assay was successfully applied to the validation of PPV removal and cleaning during SP-Sepharose cation chromatography for thrombin purification and Q-Sepharose anion chromatography for factor VIII purification. The comparative results obtained by real-time PCR assay and infectivity titrations suggested that the real-time PCR assay could be a useful method for chromatography cleaning validation and that it could have an additive effect on the interpretation and evaluation of virus clearance during the virus removal process.

• Korean Journal of Oriental Medicine
• /
• v.11 no.2
• /
• pp.155-165
• /
• 2005

This study was investigated to determine the quality control of Oriental medicine from stores dealing in Oriental medicine around Seoul and Daegu. We tested total 120 samples that widely used 15 species in herbal medicine (Lycii Fructus, Platycodi Radix, Angelicae Gigantis Radix and 12 others) being collected from Oriental medicine clinic, pharmacy, Oriental pharmacy, Oriental medical hospital and Oriental drug store. We have estimated Oriental medicine by K.P. (Korean Pharmacopoeia), K.H.P(Korean Herbal Pharmacopoeia) and announcement of KFDA. The items of examination were identification, purity, loss on drying, ash, acid insoluble ash, extract content, essential oil content, assay, heavy metal limit, and pesticides residue(BHC, DDT, Aldrin, Endrin, Dieldrin). As a result, 20 samples in total 120 samples were not satisfied with the standard and 7 species in total 15 species were not satisfied with the standard. Identification test, extract content test and pesticides residue(BHC, DDT, Aldrin, Endrin, Dieldrin) were satisfied with the standard. The result will be the basic data for the quality control of Oriental medicine.

• Research in Plant Disease
• /
• v.18 no.3
• /
• pp.231-235
• /
• 2012

Indian citrus ring spot virus (ICRSV) is known to cause a serious disease to citrus, especially to Kinnow mandarin, the popular cultivated citrus species in India. In this study, we developed diagnostic systems based on enzyme-linked immunosorbent assay (ELISA). In order to generate antibodies against ICRSV coat protein, we overexpressed the coat protein in Escherichia coli using the pET15b expression vector containing an optimized ICRSV coat protein gene. The recombinant ICRSV coat protein was overexpressed as soluble form at $37^{\circ}C$ upon IPTG induction. The protein was purified to 95% in purity by Ni-NTA column chromatography. The purified protein was immunized to rabbit for the generation of polyclonal antibody (PAb). The PAb showed a specific immunoreaction to recombinant ICRSV coat protein in western blot analysis and ELISA. Diluted rabbit antisera (10,000 fold) could detect less than 10 ng and 5 ng of the target protein in western blot and ELISA analysis, respectively.

• KSBB Journal
• /
• v.10 no.3
• /
• pp.271-282
• /
• 1995

The authentic hGH converted from met-hGH by immobilized ApM was purified by successive chromatographic processes based on the differences in isoelectric points, hydrophobicities and charges. The final recovery yield was about 14.1% and the specific activity of the purified hGH was 2.75IU per mg when assayed by enzyme immunoassay. The purified hGH was verified to be authentic hGH through the analysis of amino acid composition, amino-terminal amino acid sequence, carboxy-terminal amino acid and tryptic peptide map. The purity of purified hGH was higher than that of commercial hGH when assessed by SDS-PAGE, PAGE, IEF and HSGF. In weight-gain assay and tibia test with hypophysectomized rats, the hGH produced in this study showed the same growth effect as the commercial hGH.

• Journal of Nutrition and Health
• /
• v.53 no.6
• /
• pp.563-569
• /
• 2020

Purpose: The vascular smooth muscle cells (VSMCs) in mature animals have implicated to play a major role in the progression of cardiovascular diseases such as atherosclerosis. This study aimed at optimizing the protocol in culturing primary VSMCs (pVSMCs) from rat thoracic aorta and investigating the effect of cellular zinc (Zn) deficiency on cell proliferation of the isolated pVSMCs. Methods: The thoracic aorta from 7-month-old Sprague Dawley rats was isolated, minced and digested by the enzymatic process of collagenase I and elastase, and then inoculated with the culture Dulbecco Modified Eagle Medium (DMEM) at 37℃ in an incubator. The primary cell culture morphology was observed using phase-contrast microscopy and cellular Zn was depleted using Chelex-100 resin (extracellular zinc depletion only) or 3 µM N,N,N',N'-tetrakis(2-pyridinylmethyl)-1,2-ethanediamine (TPEN) (extracellular and intracellular zinc depletion). Western blot analysis was used for the detection of SM22α and calponin as smooth muscle cell marker proteins and von Willebrand factor as endothelial cell marker protein to detect the culture purity. Cell proliferation by Zn depletion (1 day) was measured by MTT assay. Results: A primary culture protocol for pVSMCs from rat thoracic aorta was developed and optimized. Isolated cultures exhibited hill and valley morphology as the major characteristics of pVSMCs and expressed the smooth muscle cell protein markers, SM22α and calponin, while the endothelial marker von Willebrand factor was hardly detected. Zn deprivation for 1 day culture decreased rat primary vascular smooth muscle cell proliferation and this pattern was more prominent under severe Zn depletion (3 µM TPEN), while less prominent under mild Zn depletion (Chelexing). Conclusion: Our results suggest that cellular Zn deprivation decreased pVSMC proliferation and this may be involved in phenotypic modulation of pVSMC in the aorta.

• Microbiology and Biotechnology Letters
• /
• v.50 no.3
• /
• pp.375-386
• /
• 2022

In the present study, four distinctly colored bacterial isolates that show intense pigmentation upon brief ultraviolet (UV) light exposure are chosen. The strains are identified as Micrococcus luteus (Milky yellow), Cryseobacterium pallidum (Yellow), Cryseobacterium spp. (Golden yellow), and Kocuria turfanensis (Pink) based on their morphological and 16S rDNA analysis. Moderate salinity (1.25%), 25-37℃ temperature, and pH of 7.2 are found to be the most favorable conditions of growth and pigment production for all the selected isolates. The pigments are extracted using methanol: chloroform (1:1) and the purity of the pigments are confirmed by high-performance liquid chromatography (HPLC) and thin-layer chromatography (TLC). Further, Fourier transform infrared (FTIR) and UV-Visible spectroscopy indicate their resemblance with carotenoids and flexirubin family. The antioxidant activities of the pigments are estimated, and, all the pigments have shown significant antioxidant efficacy in 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), 2,2-diphenyl-1-picryl-hydrazyl (DPPH), and ferric reducing antioxidant power (FRAP) assays. The UV protective property of the pigments is determined by cling-film assay, wherein, at least 25% of UV sensitive Escherichia coli survive with bio-pigments even after 90 seconds of UV exposure compared to control. The pigments also hold a good sun protective factor (SPF) value (1.5-4.9) which is calculated with the Mansur equation. Based on these results, it can be predicted that these bacterial pigments can be further developed into a promising antioxidant and UV-protectant for several biomedical applications.