• Title/Summary/Keyword: Isoelectric Point

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Molecular Cloning of cDNA Encoding a Putative Eugenol Synthase in Tomato (Solanum lycopersicum 'Micro-Tom') and Prediction of 3D Structure and Physiochemical Properties (토마토 'Micro-Tom' 과실의 eugenol synthase 유전자 클로닝, 단백질의 3차 구조 및 생리화학적 특성 예측)

  • Kang, Seung-Won;Seo, Sang-Gyu;Lee, Tai-Ho;Lee, Gung-Pyo
    • Journal of agriculture & life science
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    • v.46 no.4
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    • pp.9-20
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    • 2012
  • Eugenol is a volatile compound synthesized by eugenol synthase in various plants and belongs to phenylpropene compounds. However, characteristics of eugenol synthase in tomato has not been known. Therefore, we cloned a full length cDNA of a putative eugenol synthase from tomato 'Micro-Tom' using rapid amplification of cDNA ends (RACE) technique and named a clone SlEGS. Open reading frame of SlEGS was 921bp long and its deduced amino acid sequence was 307bp. The BLAST analysis indicated that SlEGS shared high similarity with PhEGS1 (67.1%) and CbEGS2 (69.4%). Amino acid composition of SlEGS was determined by CLC genomics workbench tool and 3D structure of SlEGS was constructed by homology modeling using Swiss-PDB viewer and validated using PROCHECK and ProSA-web tool. In addition, the physiochemical properties of SlEGS was evaluated using ExPASy's ProtParam tool. Molecular weight was 33.93kDa and isoelectric point was 5.85 showing acidic nature. Other properties such as extinction coefficient, instability index, aliphatic index, and grand average hydropathy was also analyzed.

Lethal (2) Essential for Life [l(2)efl] Gene in the Two-spotted Cricket, Gryllus bimaculatus (Orthoptera: Gryllidae) (쌍별귀뚜라미(Gryllus bimaculatus)의 l(2)efl cDNA 클로닝과 발현분석)

  • Kwon, Kisang;Lee, Nuri;Kwon, O-Yu
    • Journal of Life Science
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    • v.31 no.7
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    • pp.671-676
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    • 2021
  • A cDNA encoding the protein lethal (2) essential for life [l(2)efl] was cloned from Gryllus bimaculatus and named GBl(2)efl. This protein is composed of 189 amino acids, including an N-glycosylation site and 15 phosphorylation sites. Its predicted molecular mass is 21.19 kDa, with a theoretical isoelectric point of 6.2. The secondary structure of GBl(2)efl was predicted from the identification of random coils (56.08%), alpha helices (22.22%), extended strands (17.99%), and beta turns (3.7%) through sequence analyses. A homology analysis revealed that GBl(2)efl exhibited a high similarity with other species at the amino acid level, ranging from 52% to 69%. While GBl(2)efl mRNA expression was higher in the dorsal longitudinal flight muscle following a three-day starvation and in the Malpighian tubules following a one-day starvation, no changes in expression were detected in other tissues. Furthermore, tunicamycin-induced endoplasmic reticulum (ER) stress resulted in an approximately 1.8-fold higher expression in the fat body compared with the wild type.

Production of manganese peroxidase from Pleurotus ostreatus using a rotary draft tube bioreactor (RTB) and characterization of its activity (회전식 통풍관 생물반응기 사용에 따른 느타리균의 manganese peroxidase 생산 및 특성)

  • Ha, Hyo-Cheol
    • Journal of Mushroom
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    • v.19 no.4
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    • pp.316-321
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    • 2021
  • Ligninolytic enzymes were produced by Pleurotus ostreatus No.42, cultivated in a new kind of bioreactor that has a rotating draft tube with a helical ribbon. Maximum laccase (Lac) production (about 8,200 U/bioreactor) was reached after 3 days of incubation, then production decreased. Production of manganese peroxidase (MnP) in this fermenter reached a maximum level of about 8,400 U/bioreactor after 6 days of incubation. Lignin peroxidase (LiP) was not detected under these growth conditions. These results indicate that the rotary draft tube bioreactor (RTB) is compatible with large scale production of ligninolytic enzymes. MnP produced under these fermentation conditions was purified via a multistep process that included chromatography on Sepharose CL-6B, prep grade Superdex 75, and Mono-Q. This major isoenzyme was confirmed to have an apparent molecular weight of 36,400 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and its isoelectric point (IEF) was determined to be 3.95. N-terminal sequencing of the major isoenzyme from this fermentation was identical to that reported for an MnP3 isoenzyme isolated under different cultivation conditions, including stationary and shaking culture.

Functional Properties of Soy Protein Isolates Prepared from Defatted Soybean Meal (탈지대두박(脫脂大豆粕)에서 추출(抽出)한 분리대두단백(分離大豆蛋白)의 식품학적(食品學的) 성질(性質))

  • Byun, Si-Myung;Kim, Chul-Jin
    • Korean Journal of Food Science and Technology
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    • v.9 no.2
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    • pp.123-130
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    • 1977
  • A laboratory study was made to develop a simple and economic model method for the systematic determination of functional properties of 'Soy Protein Isolates (SPI)' prepared from defatted soybean meal. These are required to evaluate and to predict how SPI may behave in specific systems and such proteins can be used to simulate or replace conventional proteins. Data concerning the effects of pH, salt concentration, temperature, and protein concentration on the functional properties which include solubility, heat denaturation, gel forming capacity, emulsifying capacity, and foaming capacity are presented. The results are as follows: 1) The yield of SPI from defatted soybean meal increased to 83.9 % as the soybean meal was extracted with 0.02 N NaOH. 2) The suitable viscocity of a dope solution for spinning fiber was found to be 60 Poises by using syringe needle (0.3 mm) with 15 % SPI in 0.6 % NaOH. 3) Heat caused thickening and gelation in concentration of 8 % with a temperature threshold of $70^{\circ}C$. At $8{\sim}12\;%$ protein concentration, gel was formed within $10{\sim}30\;min$ at $70{\sim}100\;^{\circ}C$. It was, however, disrupted rapidly at $125\;^{\circ}C$ of overheat treatment. The gel was firm, resilient and self-supporting at protein concentration of 14 % and less susceptible to disruption of overheating. 4) The emulsifying capacity (EC) of SPI was correlated positively to the solubility of protein at ${\mu}=0$. At pH of the isoelectric point of SPI (pH 4.6), EC increased as concentration of sodium chloride increased. Using model system$(mixing\;speed:\;12,000\;r.p.m.,\;oil\;addition\;rate:\;0.9\;ml/sec,\;and\;temperature\;:\;20{\pm}1\;^{\circ}C)$, the maximum EC of SPI was found to be 47.2 ml of oil/100 mg protein, at the condition of pH 8.7 and ${\mu}=0.6$. The milk casein had greater EC than SPI at lower ionic strength while the EC of SPI was the same as milk casein at higher ionic strength. 5) The shaking test was used in determining the foam-ability of proteins. Progressively increasing SPI concentration up to 5 % indicated that the maximum protein concentration for foaming capacity was 2 %. Sucrose reduced foam expansion slightly but enhanced foam stability. The results of comparing milk casein and egg albumin were that foaming properties of SPI were the same as egg albumin, and better than milk casein, particularly in foam stability.

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Studies on $\alpha$-amylase of Bocillus circulans F-2 (Part II) Enzymatic characteristics of the purified $\alpha$-amylase (Bacillus circulans F-2가 생산하는 $\alpha$-amylase에 관한 연구 (제 I I 보) 정제$\alpha$-amylase의 효소적특성)

  • ;Hajime Taniguchi;Yoshiharu Maruyama
    • Microbiology and Biotechnology Letters
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    • v.10 no.2
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    • pp.123-132
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    • 1982
  • These experiments were conducted to investigate the enzymatic characteristics of the purified $\alpha$-amylase (F-2A) of Bacillus circulans F-2 and the digestion rate of various starches. 1. The molecular weight was estimated to be 93000 by SDS-polyacrylamide disc gel electrophoresis. The isoelectric point was about pH 5.0. The optimum pH for the enzyme action was 6.0-6.5 and the stable pH ranged pH 5.5-12.0. The optimum temperature was 6$0^{\circ}C$, and the purified $\alpha$-amylase was stable below 4$0^{\circ}C$. 2. The purified $\alpha$-amylase was activated by Mn$^{++}$ and Co$^{++}$, whereas it was inhibited by Ag$^{+}$, HT$^{++}$, Cu$^{++}$ and Pb$^{++}$. 3. The purified $\alpha$-amylase is considered to have no sulfhydryl residue essential for its catalytic activity. 4. Michaelis constant (Km) was 1.704 mg/$m\ell$. Activation energy between 25-4$0^{\circ}C$ was 12.297 Kcal/mole, and between 40-6$0^{\circ}C$, it was 7.831 Kcal/mole. 5. The hydrolysis product from soluble starch, amylose and amylopectin in the early stage of hydrolysis was G$_{6}$, and as hydrolysis proceeds, G$_4$and G$_2$appeared. 6. Products from each oligosaccarides are as follows: G$_4$longrightarrow G$_2$+ G$_2$,G$_3$ +G$_1$,G$_{5}$longrightarrow G$_4$+G$_1$,G$_{6}$longrightarrowG$_4$+ G$_2$,G$_{7}$ G$_4$,G$_{8}$longrightarrow G$_4$+G$_4$, 7. On raw potato starch, raw sago starch and raw yam starch, the purified enzyme exhibited a remarkably high digestion rate than Porcine pancreatic amylase and Streptococcus bovis amylase.

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Factors Related to the Development of Myocardial Ischemia During Mechanical Ventilation (인공 호흡기 적용에 따른 심근 허혈의 발생에 관한 연구)

  • Kim, Tae-Hyung;Kim, You-Ho;Lim, Chae-Man;Kim, Won;Shim, Tae-Sun;Lee, Sang-Do;Kim, Woo-Sung;Kim, Dong-Soon;Kim, Won-Dong;Koh, Youn-Suck
    • Tuberculosis and Respiratory Diseases
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    • v.46 no.5
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    • pp.645-653
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    • 1999
  • Introduction : Although myocardial ischemia tends to occur more frequently than can he documented in ventilated patients, it has not been well studied on the factors related to the occurrence of the ischemia. Methods : To investigate the related factors to ischemia development, a prospective study was done in 95 cases with consecutive 73 patients who had received mechanical ventilation(MV) in MICU. In addition to 24 h holter monitoring, echocardiogram, electrolytes, cardiac enzymes, hemodynamic, and gas exchange measurements were done within 24 h after initiation of MV in 69 cases. The measurements were repeated at weaning period in 26 cases. The ischemia was defined by the ST segment changes; up-sloping depression more than 1.5 mm or down-sloping or horizontal depression more than 1.0 mm from isoelectric baseline for 80 ms following J point. Results : Twelve patients(12.6% in 95 cases) developed ischemia in total. The incidence of ischemia development showed an increased tendency in the initial 24 hr after MV (15.9%) and in patients with left-sided heart failure found by echocardiogram (18.2%) compared with that of the weaning period (3.8%) and patients without heart failure (10.9%) (P=0.12, P=0.09, in each). There were no differences in APACHE III score, baseline ECG findings, electrolytes abnormalities, use of inotropics or bronchodilators, presence of sepsis or shock, mode of ventilation, and survival rate according to the development of ischemia. Maximal heart rates and mean arterial pressure also were not different between patients with ($137.2{\pm}30.9/min$, $82.5{\pm}15.9$ mm Hg) and without ischemia ($l29.5{\pm}29.7/min$, $83.8{\pm}17.6$ mm Hg). Conclusion : Although the incidence of myocardial ischemia was 12.6% in total, there were no clinically predictable factors to the development of ischemia during mechanical ventilation.

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Studies on Raw Silk Cohesion for Promotion (생사의 포합향상에 관한 연구)

  • 최병희;김병호;원성희
    • Journal of Sericultural and Entomological Science
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    • v.15 no.1
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    • pp.37-48
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    • 1973
  • The purpose of this studies is to improve the cohesion of raw silk through various analyses on cocoon drying, cooking, reeling, re-reeling, and on the properties of water. Also we investigated the correlation between silk testing items which we have reached to the following results. 1. Drying of cocoon When cocoons were slowly dried with 100$^{\circ}C$, the results of cohesion became much better. On the other hand, the results were considerably decreased in case the temperature with 115$^{\circ}C$. 2. Cooking of cocoon In case of the cooking of cocoon, we found that the result of cohesion was best with incomplete cooking, that of the control was next, while in over cooking, the results were very low. Also the results of cohesion were much better when using the method of over cooking with sericin arrestive agent than that of incomplete cooking with sericin agglutinating agent. 3. Reeling of cocoon A) When the temperature of reeling bath was 25-45$^{\circ}C$, the results of cohesion test were much better, but at the temperature below 25$^{\circ}C$ or above 45$^{\circ}C$, the results became worse. B) With out the process of croissieur, the results of cohesion were too bad, but in case of croissieur more than 1cm, cohesion became better rapidly. Further more, we understood that the results of cohesion were improving slightly with longer length of croissieur. C) When the velocity of reeling was increased, the results of cohesion also improved. The best results were shown when reeling velocity was 180-220 r.p.m. But when the velocity was increased more than 220 r.p.m., the results of cohesion got worse more or less. D) When the temperature of the drying pipe in reeling machine was raised, the results of cohesion also showed a tendency to improve. 4. Re-reeling A) We could net reach a conclusion as to have correlation between the number of dipping repeat in vacuum tank and the results of cohesion before re-reeling process. B) When we used Seracol 500 as an agglutination protective agent with l/1000 to l/2000 of water, the results of cohesion test were better. C) When we used Pearl-lite as an agglutination protective agent with 1/1000 to 1/2000 of water, the results of cohesion were considerably better. D) We gained tile best results when used Cohesion Improving Chemical, A-80, with 500-1500 times diluted. 1) Results of cohesion was improved when humidity was low or temperature was high in the rereeling machine. 5. Filature water A) The water pH near the isoelectric point of protein showed the best cohesion, but the farther water pH, the worser results were obtained. B) With the increasing of M-alkalinity in filature water, the results of cohesion were worse. Above all, we understood the tendency of the results of cohesion get worse when the M-alkalinity is increased above 200 ppm. C) By increasing the total hardness of the filature water, it improved the results of cohesion. Especially, when the total hardness was above 300ppm, the results were extremely high. 6. Effects combination of each results A) The result of effects combination in filature processes with the obtained best conditions was distinctively improved. But the results could not reach in mathematicaly double effect. When reelied under worse conditions, the results of cohesion test were too bad. There was "effect limit" for the promotion. B) Generally the results of cohesion were bad when the filature conditions(the temperature, pressure and the properties of water, etc) are processed as sericin loss to be high. On the other hand, the results were very good when lower sericin loss was controlled in filature conditions. C) When filature conditions such as reeling velocity and croissieur length provide pysical cohesion ability and when raw silk dry fast during reeling and re-reeling, we found the result of cohesion was better. 7. Correlation of silk testing items. A) A negative correlation exists between the results of cohesion test and cleanness defect. Another word, the result of cohesion test was found to be worse as cleanness defect increased. B) In cleanness, cohesion has negative correlation against the number of slugs, but we could not find any correlation against long loops, loose ends. C) Cohesion has negative correlation against average neatness and low neatness defect. The better the results of neatness respectively, the better the results of cohesion found. D) There is no correlation between tenacity and the results of cohesion test, but there was high positive correlation between the results of elongation and those of cohesion test. The more elongation, the better the results of cohesion was found.

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Adsorption Properties of the Lysozyme and Albumin with Physicochemical Properties of the Contact Lens (콘택트렌즈의 물리화학적 특성에 따른 라이소자임과 알부민의 흡착 특성)

  • Sung, Yu-Jin;Ryu, Geun-Chang;Jun, Jin
    • Journal of Korean Ophthalmic Optics Society
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    • v.18 no.3
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    • pp.261-270
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    • 2013
  • Purpose: Adsorption properties of lysozyme and albumin according to physiochemical properties of commercial contact lens classified with the FDA categories and a contact lens fabricated in the laboratory were investigated. Methods: The contact lens were prepared using HEMA(2-hydroxyethyl methacrylate) and TRIM(3-(trimethoxysilyl) propyl methacrylate) in a cast mold. Artificial tears containing lysozyme and albumin were prepared. We measured the amounts of protein adsorbed on the each lenses with varying adsorbed time (48 hour) and the pH range (6, 6.8, 7.4, 8.2, 9) of artificial tear. Amount of the proteins absorbed on the contact lenses were measured by using HPLC. Results: Time to reach the equilibrium of protein adsorption for silicone hydrogel lens was taken longer than hydrogel lens. The amount of adsorbed both lysozyme and albumin at equilibrium were greater for the hydrogel lens than the silicone hydrogel lens, and larger for the ionic lens than the non-ionic lens. Lysozyme was more adsorbed on the higher water content of contact lens, whereas albumin was more adsorbed on the lower water content of contact lens. Only lysozyme was adsorbed on the Group IV hydrogel lens of ionic higher water content. The adsorption of protein on contact lens increased with pH of artificial tears as close to the isoelectric point of each protein. Conclusions: The adsorption amount of lysozyme is more affected by the ionic strength of the contact lens surface than the water content of contact lens. Albumin adsorption is more affected by water content than the ionic strength of the contact lens surface. For the adsorption of proteins on the silicone hydrogel lens, the pore size, determined both by the number of Si atoms and the chemical structure of the silicone-containing monomers, as well as the polarity of contact lens should be also considered.

STUDIES ON THE EXTRACTION OF SEAWEED PROTEINS 4. Precipitation Conditions and Nutritional Evaluation of Isolated Seaweed proteins (해조단백질 추출에 관한 연구 4. 추출단백질의 심전조건 및 영양적 평가)

  • WOO Soon-Im;RYU Hong-Soo;LEE Kang-Ho
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.12 no.4
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    • pp.225-234
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    • 1979
  • For the effective utilization of diverse and abundant resource of seaweeds in Korea as a food protein supplment, extraction conditions of water, salt, and alkali soluble proteins were investigated in previous work(Ryu and Lee, 1977: Lee et al., 1977: Lee et al., 1978). The present study as a part of the serial work was thus aimed to find the conditions of isolation and purification of extracted proteins, and to evaluate the nutritional quality of the isolated seaweed proteins in terms of amino acid composition, chemical score, protein score, modified essential amino acid index(MEAAI), and in vitro digestibility presented as pepsin-pancreatin digest residue index (PPDRI). As for the isolation of extracted proteins, TCA treatment was more effective for the proteins from rhodophyceae and Chlorophyceae while the precipitation at isoelectric point was more desirable for Phaeophyceae proteins. In amino acid composition, water soluble protein fraction was superior to the other fractions in Porphyra suborbiculata whereas both water and alkali soluble fractions seemed to bo more benefitial for Enteromorpha linza and Ulva pertusa; the extraction with alcohol-alkali mixed solvent for Undaria pinnatifida and Sargassum fulvellum. Glutamic acid and aspartic acid content was particularly high in all protein fractions examined. The total amino acid content of Porphyra suborbiculata and Enteromorpha linza was almost equivalent to that of dried whole egg although the essential amino acid content was lower. A comparative analysis was made on the inedexes between raw seaweed powder and isolated protein. Chemical score of Porphyra suborbiculata and Ulva pertusa was approximately 35 and 56 in cafes of raw powder and isolated protein respectively while only 10 to 16 for raw powder of Undaria pinnatifida and Sargassum fulvellum and 30 to 35 for their isolated proteins. Protein score of all isolated proteins was in the range of 63 to 73 which indicates that isolated protein would be mere valuable than the fern of raw seaweed powder. Digestibility by means of PPDRI was found to be extremely low in case of raw powder but it could be doubled in case of isolated protein yielding 67 to 70 for Porphyra suborbiculata and Ulva pertusa.

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Effects of pH and Natural Polysaccharide Gums on the Foam Stability of Soy Protein Isolate (pH와 천연 다당류 고무질이 분리대두단백질의 거품안정성에 미치는 영향)

  • Yang, Seung-Taek;Kim, Mi-Sook;Park, Chun-Og
    • Korean Journal of Food Science and Technology
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    • v.24 no.5
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    • pp.482-491
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    • 1992
  • Experiments were conducted to elucidate the effects of pH change and sodium alginate, gum karaya and gum arabic on the foaming properties of soy protein isolate (SPI). The surface tensions of SPI solution (5%) adding to gums at pH 4.0 and 5.0, near the isoelectric point (pH 4.5) were higher than those at pH 7.0 and 8.0. Specific viscosity of the solutions adding to gums (0.1, 0.2 and 0.3%) at pH 7.0 and 8.0 were $3.6{\sim}51.8$ and $4.0{\sim}51.2$, respectively. In cases of addition of sodium alginates specific viscosity were predominantly increased, while those of gum arabic did not almost increase. Addition of gums reduced the overrun, but it stabilized the foams, especially those of sodium alginates (0.2%) increased the foam stability at pH 5.0, 7.0 and 8.0 by 57, 413 and 315%, respectively. Foaming ability, foam stability and heat stability of the foam were significantly (p<0.05) affected by changing pH. There was a tendency to increase the foaming ability according to the decrease of surface tension. Specific viscosity was also important in foam stability and heat stability.

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