• Title/Summary/Keyword: crystal protein

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Protective Effects of Ukgan-san in $CoCl_2$-induced Cell Death of C6 Glial Cells ($CoCl_2$로 유도된 C6 신경교세포의 사멸에 대한 억간산(抑肝散)의 보호 효과)

  • Cho, Mun-Young;Shin, Yong-Jeen;Ha, Ye-Jin;Woo, Chan;Kim, Ta-Jung;You, Ju-Yeon;Choi, Yong-Seok;Choi, Jung-Hoon;Shin, Sun-Ho
    • The Journal of Internal Korean Medicine
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    • v.34 no.2
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    • pp.178-191
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    • 2013
  • Objectives : In this study, we made an effort to investigate the protective mechanism of Ukgan-san (UGS) extracts on hypoxia-induced C6 glial cell death. Methods : The cell viability was assessed by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MMT) assay and cell morphological changes were analysed with microscope after staining with crystal violet (CV). Reactive oxygen species (ROS) formation was assessed by flow cytometer after staining with 2'7'-dichlorofluorescein diacetate (DCF-DA). We also analyzed expression of hypoxia-inducible factor-1 alpha (HIF-$1{\alpha}$) and p53, processing of procaspase-3 and procyclic acidic repetitive protein (PARP) by western blot method. Results : We estimated the elevated cell viability by UGS extract on $CoCl_2$-induced C6 glial cells. UGS attenuated $CoCl_2$-induced ROS formation in C6 glial cells and also showed a protective activity compared to antioxidants and exhibited abrogation of LDH-released by $CoCl_2$. UGS suppressed the typical apoptotic cell death markers, caspase-3 and PARP activation. UGS inhibited $CoCl_2$-induced HIF-1${\alpha}$ expression which is known as a major regulator for hypoxia-induced cell death, and suppressed p53 expression. Conclusions : These results suggest that UGS extract contains protective constituents for hypoxia-induced C6 glial cell death.

Post HCV Infection Due to MX Gene Stimulation Produced Post Treatment with Imported and Locally Produced Egyptian Biosimilar IFN

  • Mohamed, Shereen H;Mahmoud, Nora F;Mohamed, Aly F;Kotb, Nahla S
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.14
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    • pp.5635-5641
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    • 2015
  • Background: Cirrhosis is regarded as a possible end stage of many liver diseases, including viral infection. It occurs when healthy liver tissue becomes damaged and is replaced by scar tissue and finally may lead to hepatocellular carcinoma. Interferons (IFNs)are two general categories, type I and II. Type I includes one beta interferon and over 20 different alpha interferons. Alpha interferons are very similar in how they work, interacting with other proteins on cells like receptors. The main objective of this study was to compare Mx gene productivity post different cell line treatment with imported and Egyptian biosimilar locally produced IFNs, as well as the efficacy of those tested IFNs. Also, an assessment was made of sensitivity of different cell lines as alternatives to that recommended for evaluation of antiviral activity. Materials and Methods: Different cell lines (Vero, MDBK and Wish) were employed to evaluate cytotoxicity using the MTT assay. Antiviral activity was evaluated compared with standard IFN against VSV, Indiana strain -156, on tested rh-IFNs (imported; innovated and Egyptian biosimilar locally produced IFNs) in the pre-treated cell lines previously mentioned. The virus was propagated in the Wish cell line as recommended. Finally we estimated up-regulation of the Mx gene as a biomarker. Results: Data recorded revealed that test IFNs were safe in test cell lines. Viability was around 100%. Locally tested interferon did not realize the international potency limits, while the imported one was accepted compared with the standard IFN. These results were the same either using infectivity titer reduction assay or crystal violet staining of residual non- infected cells. Mx protein production was cell type related and confirmed by the detected Mx gene expressed in imported and locally produced IFN pre-treated cell lines. The expression of the gene was arranged in the order of Vero> wish > MDBK for the imported IFN, while for the Egyptian biosimillar locally produced one it was MDBK> Vero> wish. With regard to the antiviral activity there was a significant difference of imported IFN potency compared with the locally produced IFN (P<0.05), the IFN potential (antiviral activity) was not cell line related and showed non-significant difference for each separate product. Conclusions: Vero cells can be used as an alternative cell line for evaluation of IFN potency in case of unavailable USP recommended cell lines. Alternative potency evaluation assay could be used and proved significant difference in IFN potency in case of local and imported agents. Evaluation of antiviral activity could be used in parallel to viral infectivity reduction assay for better accuracy. Mx gene can be used as a marker for IFN potential.

Formation of Brushite (CaHPO4 · 2H2O) in the Oyster Extracts (굴 엑기스에서 결정물질 Brushite(CaHPO4 · 2H2O)의 생성)

  • Lee, Jong-Soo;Yoon, So-Mi;Jang, Jun-Ho;Lim, Chi-Won;Choi, Kwang-Yeol
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.35 no.6
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    • pp.729-733
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    • 2006
  • Small yellowish brown crystals were found in some concentrated oyster extracts which prepared by heating with the drip and washed water of boiled oyster in oyster processing factories. We collected those crystals by filtering the oyster extracts and analysed X ray diffraction (XRD), scanning electron microscopy (SEM) and minerals by ICP. Those were composed of moisture (8.1%), organic materials (21.6%) and inorganic materials (70.2%). Those major inorganic materials were determined as Ca (53.8%) and P (43.8%), from the ICP and elucidated those crystal as brushite $(CaHPO_4{\cdot}2H_2O)$ by XRD and SEM. Organic materials contained crude protein (33.7%) consisting of 7 amino acids. It is presumed that brushite may be formed by the reaction of excess Ca with P in the oyster extracts during concentrating process under higher temperature.

A Study on the Development of a Microbial Insecticide (미생물 살충제의 개발에 관한 연구)

  • Lee, Jae-Koo;Kim, Kyo-Chang
    • Applied Biological Chemistry
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    • v.19 no.4
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    • pp.189-201
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    • 1976
  • In an effort to develop a microbial in secticide, B. thdringiensis var. thuringiensis was cultured in the medium composed of cocoon-cooked water from a filature. The results obtained are summarized as followss : (1) Bacillus thuringiensis is a bacterium producing a ${\delta}-endotoxin$ especially toxic to lepidopterous insects and a thermostable exotoxin harmful to dipterous insects. (2) With a view to utilizing the cocoon-cooked water discarded from the filature, as a nutrient source in the B. thuingiensis culture, it was analyzed to contain large amounts of various minerals and protein (7.5 mg/ml) believed to be extracted from the pupae. (3) A large amount of the ${\delta}-endotoxin$ can be obtained most cheeply by using cocoon-cooked water instead of distilled water in preparing GYS and citrate salts media. (4) The largest amount of a mixture of the vegetative cells, spores, and crystals was obtained by addition of 8 gr/l of glucose to the GYS medium. (5) The growth of the bacterium was far better, when leucine, isoleucine, and valine were added all together to the citrate salts medium to the concentration of $1.25{\times}10^{-3}M$. (6) The best growth was observed by addition of Na-glutamate to the citrate salts medium to the concentration of $2.5{\times}10^{-3}M$. (7) The optimal culture time ranged from 9 to 15 days. (8) The highest mortality was shown in Pieris rapae Linne with a pH of the total body extract of 8.4, whereas Dendrolimus spectabilis Butler and Bombyx mori Linne with lower pH's were less susceptible to the ${\delta}-endotoxin$. (9) The presence of the thermo stable exotoxin was confirmed by the fact that the supernatant of the culture was very toxic to the Drosophila melanogaster tested.

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Homology Modeling and Characterization of Oligoalginate Lyase from the Alginolytic Marine Bacterium Sphingomonas sp. Strain MJ-3 (알긴산을 분해하는 해양미생물인 Sphingomonas sp. MJ-3 균주의 올리고알긴산 분해효소의 상동성 모델링 및 특성연구)

  • Kim, Hee Sook
    • Journal of Life Science
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    • v.25 no.2
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    • pp.121-129
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    • 2015
  • Alginates are found in marine brown seaweeds and in extracellular biofilms secreted by some bacteria. Previously, we reported an oligoalginate lyase from Sphingomonas sp. MJ-3 (MJ3-Oal) that had an exolytic activity and protein sequence homology with endolytic polymannuronate (polyM) lyase in the N-terminal region. In this study, the MJ3-Oal was tested for both exolytic and endolytic activity by homology modeling using the crystal structure of Alg17c from Saccharophagus degradans 2-40T. The tyrosine residue at the $426^{th}$ position, which possibly formed a hydrogen bond with the substrate, was mutated to phenylalanine. The FPLC profiles showed that MJ3-Oal degraded alginate quickly to monomers as a final product through the oligmers, whereas the Tyr426Phe mutant showed only exolytic alginate lyase activity. $^1H$-NMR spectra also showed that MJ3-Oal degraded the endoglycosidic bond of polyM and polyMG (polymannuronate-guluronate) blocks. These results indicate that oligoalginate lyase from Sphingomonas sp. MJ-3 probably catalyzes the degradation of both exo- and endo-glycosidic bonds of alginate.

Studies on Some Physico-Chemical Properties of the Acorn Starch (도토리전분(澱粉)의 이화학적(理化學的) 성질(性質)에 관(關)한 연구(硏究))

  • Kim, Jung-Ok;Lee, Mahn-Jung
    • Korean Journal of Food Science and Technology
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    • v.8 no.4
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    • pp.230-235
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    • 1976
  • The current experiment has been carried out in order to find the chemical and physical properties of three species of the acorn starch. The results of the experiment are as follows. 1. Acorn is made up of 3.5-5.0% of lipid, 7.2-7.7% of protein, 71.6-76.3% of carbohydrate, and 1.9-2.2% of ash. 2. The purifying method of ether treated is better than that of 85% methanol treated for isolating pure starch. 3. Initial gelatinization temperature of the acorn starches ranged from $61-68^{\circ}C$; Quercus crispula Blume was the lowest and Quercus mongolica Fischer was the highest. On viscosity, Quercus mongolica Fischer shows the highest and Quercus crispula Blume shows the lowest. 4. It is estimated that the acorn starch lie halfway between those of the potato starch and the corn starch. By drying at $100^{\circ}C$, the crystal structure is destroyed first in the farther part of the lattice distance.

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The Effect of Jeju Wild Ginseng Extracts on Skin Barrier via Serine-Palmitoyltransferase (제주산양산삼이 세린-팔미토일 전이효소(Serine-Palmitoyltransferase)를 통해 피부 장벽에 미치는 효과에 대한 연구)

  • Kim, Hyo Min;Lee, Jung No;Kim, Jae Moon;Kim, Sung Kyu;Park, Sung-Min
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.42 no.2
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    • pp.119-126
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    • 2016
  • Skin is the largest organ that protects the body from the external environmental factors such as smog, cigarette smoke, UV. Protective skin barrier is composed with keratinizational keratinocytes and intercellular lipids such as ceramides, cholesterols and fatty acids combined by the lamellar liquid crystal structure. In this research, we confirmed that the Jeju wild ginseng (JWG) extracts dose-dependently increased the expression of serine-palmitoyltransferase (SPT) protein which is associated with ceramide biosynthesis. In addition, emulsion containing 5% JWG extract was applied on skin of human volunteers for 2 weeks and then significantly reduced transepidermal water loss (TEWL) compared to that of control group. As a results, JWG extract increased the biosynthesis of ceramides that is the key components of the skin lipid through enhancing expression of SPT. In addition, JWG extract reduced TEWL resulting in improvement of skin barrier function. In this context, we suggest that JWG extract could be used as a skin barrier enhancer and moisturing agents in cosmetic fileds.

Biosynthesis of the extracellular enzymes in de novo during the differentiation of Aspergillus niger (검정곰팡이의 형태분화에 따른 세포외성효소의 신생적생합성에 관한 연구)

  • Kim, Jong-Hyup
    • The Korean Journal of Mycology
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    • v.6 no.2
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    • pp.1-10
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    • 1978
  • In de novo biosynthesis of the extracellulor enzymes-proteinsaes, alpha and gluc-amylases during the synchronized differentiation of Aspergillus niger in submerged culture and surface liquid culture were investigated. Gluc-amylase was synthesized in the stage of presporulation in which phialide formation is involved. Proteinase was synthesized both in the stages of conidiophore formation and presporulation. Alpha-amylase was synthesized during presporulation and sporulation stages, the activity of enzyme lasted for seven days on surface liquid culture. It seemed that the synthesis was occured in de novo partly repressed by the catabolite, and its nature was found to be constitutive since it is produced in non-starch medium. Polyacrylamide gel electrophoresis have shown that presporulating and sporulating body produced diverse types of the proteins whereas the earlier stages of vegetative body showed simpler profiles. The uptake of C-14 uracil into RNA and C-14 glutamate into protein were shown to be vigorous in presporulating body rather than those in sporulating body. Coincidence of alpha-amylase biosynthesis in de novo and sporulation may be significant in the study of differentiation in which gene expression is involved.

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Studies on Anisakis Type Larvae (Anisakis 형(型) 유충(幼蟲)에 관한 연구(硏究))

  • Lim, Jung Teck
    • Korean Journal of Veterinary Research
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    • v.15 no.2
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    • pp.293-307
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    • 1975
  • As it has been known recently that anisakis type larvae harbouring in marine fishes are a causal agent of zoonosis to human and probably to land living mammal animals, attention was focused on the study on the larvae in an aspect of epidemiology or epizootiology. The present work was conducted from 1966 to 1975 for i) survey on the harbouring status of anisakis type larvae in marine fishes of this country, ii) observation on the response to the experimental infestation of the larvae to the pigs, in the reason that they could well fetid raw fish viscera occasionally containing the larvae as a high protein source of swine food, and iii) observation on the larval resistance and response to vermicidal agents for the purpose of prevention of the larval infection to the mammal animals. The data obtained in the studies were summarized as follows: 1. In the survey on the status of larvae harbouring in main species of marine fishes of this country, 15 species, a total of 1,940 fishes, were observed and the result was summarized in table 2. Average number of larvae, in upper rank of 5 out of all 15 species of fishes, were as highest as 156 larvae ranging 74 to 450 in Pseudosciaena manchurica (chamjogi), 54.5 ranging 15 to 240 in Trichiurus haumela (kalchi), 35.6 ranging 8 to 112 in Trachurus japonica (junggengi), 30.6 ranging 4 to 65 in Parapristipama trilineatum (benjari) and 20.5 ranging 3 to 48 in Nibea argentata (boguchi) respectively. In morphological observation, size of the larvae in the fishes were varied, ranging from 2 to 32mm long, and a tendency to larger size and number of larvae in the fishes, which were wider sea migration, higher age and lager bodily size, was observed The favorite places harbouring the larvae in fishes were mainly around the intraperitoneal viscera such as mesentery, omentum, liver, pyloric suspensory, fat tissue and cloaca, and rarely in body muscles of fish. Fishes heartily infested with the larvae showed stunted growth decreased egg formation and severe damage of liver. 2. In the experimental infestation of the larvae to normal pigs, as illustrated in table 3, a group with large dose of larvae (a total of 1,800 larvae, 300 larvae Per dose, twice in a dart for 3 days) showed acute clinical syndrome terminatine death with a week course, whereas two groups with less dose of larvae (a total of 180~360 larvae, 10 larvae per dose, at 5 days interval for 70~180 days) showed subclinical syndrome with remarkably stunted growth as. much as approximately one half of body size in contest to the control pigs. In the pathological findings, a group with large dose of larvae showed macroscopically larvae penetrating to the gastric wall with severe gastroenteritis, and histopathologically various acute lesions caused by active larvae penetration into the wall of stomach and interstine, whereas two groups with less dose of larvae showed chronic lesions such as hypertrophy and verminous granulomatous swelling of gastric wall, suggesting strongly the possibility of natural infestation of larvae to swine. 3. In the resistance of the larvae to the chemical solutions, the larvae tolerated for 2 days in 15 percent solution of sodium chloride and acetic acid, and for 7 days in 70 percent solution of ethyl alcohol. In the resistance to the temperature, the larvae died within 1 second at $62^{\circ}C$ and tolerated for 24 hours at $-3^{\circ}C$, 12 hours $-5^{\circ}C$ respectively. 4. For the experiment on the vermicidal effect to larvae, general vermicidal drugs such as Neguvon, Combantrin, antimony Potassium, piperazine adipate and piperazine dihydrochloride, oxidizer such as potassium permanganate and potassium chlorate, and dyes such as gentian violet and crystal violet were used, and among them, as illustrated in table 6, potassium permanganate was proved as the best. In the successive test for the practical use of potassium permanganate, vermicidal effect in seawater solution of potassium permanganate and common-water solution of potassium permanganate were compared, and then retested by dipping the fish viscera including the larvae into the two different solutions of potassium permanganate. The result through these tests indicated that 0.01 percent common water and sea-water solution of potassium permanganate could be apparently recommended as a preventive vermicidal solution, having 90 to 100 percent vermicidal effect by dipping for 12 to 24 hours even though sea-water solution of potassium permanganate had a tendency to slightly less effect than the common-water solution of potassium permanganate (Table 8).

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Assessment of Bio-corrosive Effect and Determination of Controlling Targets among Microflora for Application of Multi-functional CFB on Cement Structure (다기능 탄산칼슘 형성세균의 시멘트 건축물 적용위한 부식능 평가 및 건축물 정주미생물 중 방제 대상 결정)

  • Park, Jong-Myong;Park, Sung-Jin;Ghim, Sa-Youl
    • Journal of Life Science
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    • v.25 no.2
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    • pp.237-242
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    • 2015
  • The use of calcite-forming bacteria (CFB) in crack remediation and durability improvements in construction materials creates a permanent and environmentally-friendly material. Therefore, research into this type of application is stimulating interdisciplinary studies between microbiology and architectural engineering. However, the mechanisms giving rise to these materials are dependent on calcite precipitation by the metabolism of the CFB, which raises concerns about possible hazards to cement-based construction due to microbial metabolic acid production. The aim of this study was to determine target microorganisms that possibly can have bio-corrosive effects on cement mortar and to assess multi-functional CFBs for their safe application to cement structures. The chalky test was first used to evaluate the $CaCO_3$ solubilization feature of construction sites by fungi, yeast, bacterial strains. Not all bacterial strains are able to solubilize $CaCO_3$, but C. sphaerospermum KNUC253 or P. prolifica KNUC263 showed $CaCO_3$ solubilization activity. Therefore, these two strains were identified as target microorganisms that require control in cement structures. The registered patented strains Bacillus aryabhatti KNUC205, Arthrobacter nicotianae KNUC2100, B. thuringiensis KNUC2103 and Stenotrophomonas maltophilia KNUC2106, reported as multifunctional CFB (fungal growth inhibition, crack remediation, and water permeability reduction of cement surfaces) and isolated from Dokdo or construction site were unable to solubilize $CaCO_3$. Notably, B. aryabhatti KNUC205 and A. nicotianae KNUC2100 could not hydrolyze cellulose or protein, which can be the major constituent macromolecules of internal materials for buildings. These results show that several reported multi-functional CFB can be applied to cement structures or diverse building environments without corrosive or bio-deteriorative risks.