• Title/Summary/Keyword: Protein pattern

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IMMUNOHISTOCHEMICAL ASSAYS FOR THE EXPRESSION OF EPIDERMAL GROWTH FACTOR-SIGNALING PROTEINS IN ADENOID CYSTIC CARCINOMAS OF HUMAN SALIVARY GLANDS (타액선 선양낭성암종에서 상피성장인자 신호전달 단백의 발현에 관한 면역조직화학적 연구)

  • Park, Young-Wook;Kim, Jung-Hwan
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.28 no.6
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    • pp.499-510
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    • 2006
  • Malignant tumors of the human salivary glands may arise from major or minor salivary glands. Adenoid cystic carcinoma (ACC) is the second most common malignant neoplasm in the salivary glands. ACC is occasionally highly aggressive tumor that readily invades adjacent tissues and metastasize to distant organs at early stages of the disease. Although ACC tends to grow slowly, treatment outcome may be poor due to wide local infiltration, perineural or intraneural spread and a propensity for hematogenous metastasis. Therefore, knowledge of cellular and molecular characteristics that influence the growth, survival and metastasis of tumor cells, is important for new treatment strategies of salivary ACC. I determined expressions of epiderma growth factor (EGF)-signaling molecules using surgical specimens of human ACCs. Protein expressions of EGF, transforming growth $factor(TGF)-{\alpha}$, EGF receptor (EGFR), phosphorylated EGFR (pEGFR), and human EGF receptor (HER)-2 were assessed in 18 cases of salivary ACC by immunohistochemical staining. Adjacent normal salivary tissues and mucosal tissues, uninvolved by the malignant tumor, served as internal controls. Most of the tumors, especially ACC with a tubulocribriform pattern, were positive for EGF signaling molecules. The overall percentages of the 18 specimens expressing EGF, $TGF-{\alpha}$, EGFR, pEGFR, and HER2 were 50, 89, 61, 61 and 83% respectively. Moreover, tumor-associated endothelial cells and infiltrating immune-related cells in the stroma of ACC, also expressed these biomarkers. Taken together, EGF-signaling molecules are actively expressed in salivary ACC. Therefore, we suggest that these biomarkers can be molecular targets for new treatment strategies of salivary tumors.

Effects of Seed Soaking of Kinetin on Alleviating Copper Toxicity during Germination in Rice (Kinetin 침종처리가 벼 발아중 구리 독성 경감에 미치는 영향)

  • 김상국;이상철;민기군;이승필;최부술
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.41 no.4
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    • pp.465-474
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    • 1996
  • The study was carried out to determine the proper concentration of plant growth regulator, kinetin on alleviating copper toxicity for two rice cultivars of seed germination. The results were as followings : Soaking treatment of kinetin 10$^{-3}$ M increased the germination rate of both cultivars, Ilpumbyeo and Hyangmibyeo 1 by 92% and 88% as compared with copper treatment (60ppm). But the soaking treatment effect of plant growth regulator, kinetin was not recognized at the kinetin 10$^{-4}$ M and 10$^{-5}$ M. Chlorophyll content of both rices was higher than that of Hyangmibyeo 1. Copper content of Ilpumbyeo was higher in leaf than in seed part. At the 3 days after treatment of copper 60ppm, both cultivars of treatment of kinetin 1O-3M showed the somewhat thin bands at the 35 and 40kDa compared with others. A new protein band pattern was only appeared to kinetin 1O-3M at approximately 54.4kDa(M. W) at the 7 days after treatment of copper 60ppm in llpumbyeo cultivar, SOD activity of copper 60ppm treatment increased in 3DAT, but there were no significant differences in 5 and 7DAT of two cultivars. Free proline contents of copper 60ppm treatment in llpumbyeo were remarkably increased about 4.996$\mu$M. In particular, free proline content of kinetin l0$^{-3}$ M in Ilpumbyeo was 5.008$\mu$M in 3DAT. In case of Hwangmibyeo 1, free proline content of copper 60ppm was 5.825$\mu$M compared with an untreated control showing 2.34l$\mu$M. The effects of kinetin treatment were recognized to promote the root growth and germination rate under copper toxicity(60ppm) condition in both cultivars.

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Disturbance of $\alpha$-Amylase Secretion from Bacillus amyloliquefaciens Cells by the Treatment of Puromycin and Magnesium (Bacillus amyloliquefaciens에서 Puromycin 과 Magnesium에 의한 $\alpha$-Amylase 의 분비저해)

  • 안순자;김순옥;이동희;송방호
    • Microbiology and Biotechnology Letters
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    • v.17 no.5
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    • pp.412-420
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    • 1989
  • To know how the ribosomes involved in secretory protein synthesis were attached to the cytoplasmic membrane in Bacillus amyloliquefaciens, the cells were treated with puromycin combinated with magnesium at the logarithmic phase, and the variation of cell-bound and extracellular $\alpha$-amylase activity was assayed for determining the $\alpha$-amylase translocation blocking through the cytoplasmic membrane. In the abnormal $\alpha$-amylase producing mutant in which the C-terminal of the $\alpha$-amylase structure was deleted, B. umytotiquefaciens CH10-2, the $\alpha$-amylase was translocated normally through the cytoplasmic membranes, and the translocation blocking by puromycin was revealed to have a similar pattern as that in the wild type. This means that the C-terminal part of the enzyme structure may not have a signal for secretion. The cell death of the logarithmic phase cells in both strains was not affected much under 20$\mu\textrm{g}$/$m\ell$ of puromycin, however, the $\alpha$-amylase translocation was blocked markedly under less than 10$\mu\textrm{g}$/$m\ell$ of the puromycin concentration. The blocking of the enzyme secretion by puromycin may be due to the detachment of the ribosomes from cytoplasmic membranes by disturbing the nascent polypeptide synthesis. Further evidence for confirming this was that the detachment was increased in 50 mM of magnesium ion because the extracellular $\alpha$-amylase activity was decreased more under this condition. If the cells were treated with trypsin combinated with Iysozyme, the extracellular $\alpha$-amylase activity from the cultured medium was reduced markedly, however, the activity from the cells treated with trypsin only was not reduced. This means that the nascent polypeptides protruding from the cytoplasmic membrane were sensitive to the trypsin digestion, whereas the matured ones were not. Therefore, the protruding polypeptides from the cytoplasmic membranes may be truncated by trypsin before forming their final tertiary structures by folding in the cell wall layer.

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On-site Output Survey and Feed Value Evaluation on Agro- industrial By-products (농산업부산물들에 대한 배출 현장 조사 및 사료적 가치 평가)

  • Kwak, W. S.;Yoon, J. S.
    • Journal of Animal Science and Technology
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    • v.45 no.2
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    • pp.251-264
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    • 2003
  • This study was conducted to make on-site survey on the output pattern and utilization situation of 19 by-products selected, to evaluate their nutritional characteristics, to find out a reliable index with which digestion of by-products can be predicted on the basis of chemical compositions analyzed and to diagnose the risk of using book values in the absence of the actual values analyzed for diet formulation. Production and utilization situations of by-products were quite various. Nutritionally, fruit processing by-products such as apple pomace (AP), pear pomace (PP), grape pomace (GP), and persimmon peel (PSP), and bakery by-products (BB) were classified as energy feeds. Soybean curd meal (SCM), animal by- products such as blood (BD), feather meal (FM) and poultry by-products (PB), and activated milk processing sludge (AMS) were classified as protein feeds. Soy hulls (SH), spent mushroom compost (SMC), barley malt hulls (BMH), waste paper (WP) and broiler litter (BL) were classified as roughage. Rumen contents (RC) and restaurant food waste (FW) were nutritionally analogous to complete diets for cattle and swine, respectively. Compared to soybean meal (SBM), BD and FM contained high (P<0.05) levels of amino acids and barley malt sprouts (BMS), AMS and FW contained low (P<0.05) levels of amino acids. Enzymatic (pepsin) digestibilities of proteinaceous feeds ranged between 99 and 66%. In vitro DM digestibility was high (P<0.05) in the order of FW, BB, AP, SH, PP, PSP, BMH, BMS, SCM, GP, RC, PB, BL, WP, SMC, AMS, FM and BD. In vitro DM digestibility had the highest correlation (r=0.68) with nonfibrous carbohydrate among chemical components. Differences between analyzed values of chemical components and book values were considerable. Caution is required in using book values when large amount of by-products are used in diets.

비만(肥滿) CLINIC 내원환자(來院患者) 453 CASES에 대(對)한 임상적(臨床的) 고찰(考察)

  • An, Gyeong-Sun;Seong, Nak-Gi
    • Journal of Haehwa Medicine
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    • v.2 no.2
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    • pp.219-246
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    • 1993
  • In 1991, Obesity rate of South Korea has reached to 18.7%. Because of economical development, the pattern of diet is exchanged from carbohydrate to rich protein and fat. The more problem is not only obesity of adult but also one of little child. Obesity is induced to diabetes mellitus, hypertension, artherosclerosis, hyperlipoidemia. heart and C.V.A disease, etc. In Woman, special important ploblem is the complex of beauty about Woman's figure. In Oriental Medicine, the factor of obesity is mainly regarded as dampness. And there are many treatments and methods to body weight loss, but obesity patients dislike to use them because of their side effects and inconvenience, intolerance. Now ear acupuncture is applied on so many disease because of its easy handly, non-side effect and high efficiency in clinics. Here obesity acupuncture is used to ear and whole body acupuncture. Because they react eachother for lack point. Therefore, in order to investigate the effect of obesity acupuncture and develop non-drug, non-starvation etc, we analyzed 453 the cases of body weight loss patients treated with ear and whole body acupuncture in Oriental Medicine Hospital of Jeon-Ju Woo-Sug University from April.1.1992. to March.17. 1993. The results were summarized as follows. 1. Distribution of sex ; male (4.4%), Female(95.6%) 2. Distribution of age in descending order ; 30s, 20s, 40s, 10s, 50s, below 10s, abowe 60s. The 20s-30s are group made up 60.7% of the group. 3. Distribution of occupation in descending order; housewife, student, service, salaried, merchant, teacher, farmer, inoccupation. 4. Distribution of human coporal constitution in descending order : Tae-Eum-In, So-Eum-In, So-Yang-In. 5. Distribution of body height and weight, 155-164cm ; 71.1%, 60-70kg, 74.6% are majority. 6. Distribution of weight variation, 2-6kg(71.0%) is majority, also 13-14kg(0.4%). 7. Distribution of duration in descending order ; 1-3 years, 3-6 years, 1-12months, above 10 years but in success, 1-12 months, 1-3 years, 3-6 years, above 10 years. Therefore, we know that the shorter duration of obesity is, the more loss of body weight. 8. Past experiences to body weight loss; Yes(69.5%), No(30.5%). The success rate accordant with the past temporary experiences shows that the cases without experience is higher than the ones with experience. 9. In distribution of times(treatments), 10 times is top. The rate of body weight loss is the highest in 14 times. Therefore, I think that one would need at least 10 times. in order lose body weight 10. Distribution of body weight variation in treatments times is at 2 times(3-4kg loss), and surprisingly is 14kg loss at above 15 times. 11. Distribution of symptoms improvement, in descending order ; heavy sense in body, dec. of appetite, inc.of exercise, lumbago, edema, knee pain, inc.of urine, inc. of fullness sense, thirsty, disease of gynecology, white tung, chest burning, heart burning, dec.of tobacco, drink taste. motion sickness, allergy, water eczema, arthma, belching. 12. Distribution of snack; Yes(87.4%), No(78.6%) 13. Distribution of exercise; Yes(21.4%), No(78.6%) 14. Distribution of sleeping times, above 7 hours(79.0%) 15. Distribution of the reason to body loss, the complex of beauty(68.7%) is top. 16. Distribution of side effect in obesity acupuncture, constipation (17.4%) is top. 17. Distribution of method in body weight loss ; dietary treatment (31.1%), sauna(26.7%), exercise(19.7%), the center of body weight loss (15.0%) herb-med and starvation treatments (5.1%), hand-finger acupuncture (hand-foot acupuncture) is 1.6%, diet pill(0.3%), etc(0.6%).

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THE EXPRESSION OF MSX GENES DURING EARLY CRANIAL SUTURE EMBRYOGENESIS (두개골 봉합부의 초기 형태발생과정에서 Msx 유전자들의 발현양상)

  • Lee, Sang-Youp;Park, Mi-Hyun;Ryoo, Hyun-Mo;Nam, Soon-Hyeun;Kim, Young-Jin;Kim, Hyun-Jung
    • Journal of the korean academy of Pediatric Dentistry
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    • v.30 no.1
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    • pp.171-180
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    • 2003
  • The development of calvarial bones is tighly co-ordinated with the growth of the brain and needs of harmonious interactions between different tissues within the calvarial sutures. Premature fusion of cranial sutures, known as craniosynostosis, presumably involves disturbance of these interactions. Mutations in the homeobox-containg gene Msx2 cause human craniosynostosis syndrome. Msx genes, which are consist of Msx1, Msx2 and Msx3, are homeobox-containg transcripton factors, and were originally identified as homologue of Drosophila msh(muscle segment homeobox) gene. Msx1 and Msx2 genes, expressed mostly in overlapping patterns at multiple site of tissue interactions during vertebrate development, are associated with epithelial-mesenchymal interactions during organogenesis, targets of BMP and FGF signaling. To elucidate the function of Msx genes in the early morphogenesis of mouse cranial suture, we analyzed the expression of them by in situ hybridization during embryonic(E15-E18) stage, and did vivo experiments in E15.5 mouse using rhBMP-2, rhFGF-2 protein soaked bead. In the sagittal suture, Msx1 was expressed in the mesenchyme of suture and the dura mater, Msx2 was intensely expressed in the sutural mesenchyme and the dura mater. In the coronal suture both of Msx genes were expressed intensely in the sutural mesenchyme and expressed in the periosteum also. Msx1 had a broader expression pattern than Msx2. BMP2 beads induced expression of both Msx1 and Msx2, FGF2 beads induced expression of Msx1, but not Msx2. Taken together, these data suggest that Msx1 and Msx2 genes have important role in regulating the morphogenesis and maintenance of embryonic cranial suture. Both of Msx genes are expressed similarly but because of their upstream signaling, they function dependently or cooperatively according to change of signaling molecule.

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Study of Hedyotis Diffusa Methanol Extract on Anti-tumoral Effect and Mechanism (백화사설초(白花蛇舌草) 메탄올 추출물(抽出物)의 항종양(抗腫瘍) 효과(效果) 및 항암(抗癌) 기전(機轉)에 관(關)한 연구(硏究))

  • No, Hoon-Jeong;Moon, Gu;Moon, Seok-Jae;Won, Jin-Hee;Moon, Young-Ho;Park, Rae-Gil
    • THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY
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    • v.6 no.1
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    • pp.81-97
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    • 2000
  • Objectives: This experimental study was carried out to evaluate the effects of aqueous and methanol extracts of Hedyotis diffusa which has long been used for cancer treatment in oriental medicines on the induction of apoptotic cell death in human lymphoid leukemia cell line, HL-60. Methods: Cells were treated with various concentrations (200 to $0.4{\mu}g$) and periods (6 to 30 hr) of $H_2O$ and methanol extracts of Hedyotis diffusa. Then, cells were tested for viability by MTT assay. Cells wrere treated with $200{\mu}g/ml$ of methanol extract fork various periods. Genomic DNA was isolated, separated, on 1.5% agarose gels, stained with ethidium bromide and visualized under UV light. Cells were treated with $200{\mu}g/ml$ of each extract for 16 hr. Then, cells were treated with Hoechst dye 33342 and observed by fluorescence microscopy. Cells were treated with various doses of each for 12 hr and $100{\mu}g/ml$ of methanol extract for various periods. Lysate from the cells used to measure the activity of Caspase-1 and-3 proteases by using fluorogenic peptide substrates including acetyl-YVAD-AMC and acetyl-DEVD-AMC, respectively. Cells were treated with $200{\mu}g/ml$ of each extract for various periods. Cell lysates were immunoprecipated with anti-JNKl antibodies. The immune complex was reacted with $32^p-ATP$ and c-Jun as a substrate. The phosphotransferase activity of JNKI was measured by using PhosphoImage analyzer (Fuji Co., Japan). Nuclear extracts were isolated and incubated with oligonucleotide probe of $NF-{\kappa}B$. Transcriptional activation of ${\kappa}B$ was measured by using EMSA and visualized by PhosphoImage analyzer (Fuji Co, Japan). Cell lysates were prepared and analyzed by Western blotting with anti-Bc12 antibodies and anti-Bax antibodies. Cells were pretreated with various doses of methanol extract for 2 hr. Then, the extract was removed by centrifugation. Cells were resuspended with RPMI-1640 media containing 0.3% agarose, 10% FBS, overlayred onto bottom layer agarose and incubated at $CO_2$ incubator for 6 days. The number of colony was counted under light microscopy ($\time100$). Results: The death of HL-60 cells was markedly induced by the addition of methanol extract of Hedyotis diffusa in a dose and time-dependent manners. The apoptotic characteristic ladder pattern of DNA strand break was observed in death of HL-60 cells. In addition, it was shown nucleus chromatin condensation and fragmentation under Hoechst staining. Therefore, Hedyotis diffusa extract-induced death of HL-60 cells is mediated by apoptotic signaling processes. The activity of Caspase 3-like proteases remained in a basal level in HL-60 cells treated with aqueous extract of Hedyotis diffusa. However, it was markedly increased in HL-60 cells treated with methanol extract of Hedyotis diffusa. In addition, the phosphotransferase activity of JNKl was increased in HL-60 cells treated with methanol extract of Hedyotis diffusa. Furthermore, the activation of transcriptional activator, $NF-{\kappa}B$ was markedly induced by methanol extract of Hedyotis diffusa. Anti-apoptotic Bc12 was cleaved into 23Kda fragment by treatment of methanol extract of Hedyotis diffusa. However, expression of proapoptotic Bax protein was increased by treatment of methanol extract of Hedyotis diffusa in a time-dependent manner. Furthermore, methanol extract markedly inhibited the colony forming efficiency of HL-60 cells in semisolid agar culture. Conclusions: Above results suggest that methanol extract of Hedyotis diffusa induces the apoptotic death of human leukemic HL-60 cells via activations of Caspase-3 proteases, JNKI, transcriptional activator $NF-{\kappa}B$, In addition, our results also suggest that methanol extract of Hedyotis diffusa reduces the malignant potential of HL-60 cells via down regulation of colony forming effciency through cleavage of Bc12 as well as induction of Bax.

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Identification of the Pig β-1,3-N-acetylglucosaminyltransferase 1 (pB3GNT1) that is Involved in Poly-N-acetyllactosamine (poly-LacNAc) Synthesis (Poly-N-acetyllactosamine (poly-LacNAc) 합성에 관여하는 돼지 β-1,3-N-acetylglucosaminyltransferase I (pB3GNT1) 유전자 동정)

  • Kim, Ji-Youn;Hwang, Hwan-Jin;Chung, Hak-Jae;Hochi, Shinichi;Park, Mi-Ryung;Byun, Sung June;Oh, Keon Bong;Yang, Hyeon;Kim, Kyung-Woon
    • Journal of Life Science
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    • v.28 no.4
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    • pp.389-397
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    • 2018
  • The structure of glycan residues attached to glycoproteins can influence the biological activity, stability, and safety of pharmaceutical proteins delivered from transgenic pig milk. The production of therapeutic glycoprotein in transgenic livestock animals is limited, as the glycosylation of mammary gland cells and the production of glycoproteins with the desired homogeneous glycoform remain a challenge. The ${\beta}$-1,3-N-acetylglucosaminylatransferase1 (B3GNT1) gene is an important enzyme that attaches N-acetylglucosamine (GlcNAc) to galactose (Gal) residues for protein glycosylation; however, there is limited information about pig glycosyltransferases. Therefore, we cloned the pig B3GNT1 (pB3GNT1) and investigated its functional properties that could attach N-acetylglucosamine to galactose residue. Using several different primers, a partial pB3GNT1 mRNA sequence containing the full open reading frame (ORF) was isolated from liver tissue. The ORF of pB3GNT1 contained 1,248 nucleotides and encoded 415 amino acid residues. Organ-dependent expression of the pB3GNT1 gene was confirmed in various organs from adult and juvenile pigs. The pB3GNT1 mRNA expression level was high in the muscles of the heart and small intestine but was lower in the lungs. For functional characterization of pB3GNT1, we established a stable expression of the pB3GNT1 gene in the porcine kidney cell line (PK-15). As a result, it was suggested that the glycosylation pattern of pB3GNT1 expression in PK-15 cells did not affect the total sialic acid level but increased the poly N-acetyllactosamine level. The results of this study can be used to produce glycoproteins with improved properties and therapeutic potential for the generation of desired glycosylation using transgenic pigs as bioreactors.

Physicochemical Properties of Fish-meat Gels Prepared from Farmed-fish (해수어를 활용한 연제품의 제조 및 물리화학적 특성)

  • Kim, Hyung Kwang;Kim, Se Jong;Karadeniz, Fatih;Kwon, Myeong Sook;Bae, Min-Joo;Gao, Ya;Lee, Seul-Gi;Jang, Byeong Guen;Jung, Jun Mo;Kim, Seo yeon;Kong, Chang-Suk
    • Journal of Life Science
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    • v.25 no.11
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    • pp.1280-1289
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    • 2015
  • Fish-meat gel is being produced mostly relying on surimi and raw materials imported from Southeast Asia and North America and present in small amount in local markets. In this study, common farmed local fishes were examined as stable and reliable sources of surimi for fish-meat gel production. For testing, five main farmed-fish of Korea, namely; Bastard halibut (Paralichthys olivaceus), Red sea bream (Pagrus major), Korean rockfish (Sebastes schlegeli), Common mulle (Mugil cephalus), and Finespotted flounder (Pleuronichthys cornutus) were used following a traditional washing process. The quality of the surimi was determined by the values of water content, whiteness index, gel strength and impurity. Accordingly, fish-meat gel and surimi quality experiments were carried out by measuring compressive and texture properties, expressible moisture content, Hunter color scale values and SDS-page protein patterns. Also gel characteristics were compared with that of FA and RA grade surimi (Alaska Pollock). Fish-meat gels were prepared by salt mincing the farmed-fish surimi with NaCl (2% w/w) and moisture adjustment to 84% by ice water adding. Prepared fish-meat paste was filled into 20-25 cm long polyvinylidene chloride casings and heated at 90℃ for 20 min. The whiteness values of fish-meat gels produced from surimi were increased by using farmed-fish and became comparable to that of FA Alaska Pollock gel. Among all tested farmed-fish, P. olivaceus and P. major exhibited better properties than RA Alaska Pollock and similar properties to FA Alaska Pollock. Therefore, current data suggests that fish farming can be an efficient and sustainable fish-meat source for fish-meat gel production in Korea.

Starch Structure and Physicochemical Properties of Colored Rice Varieties (유색미 품종별 전분 구조 및 이화학적 특성)

  • Park, Ji-Young;Oh, Sung-Hwan;Han, Sang-Ik;Lee, Yu-Young;Lee, Byung-Won;Ham, Hyeonmi;Choi, Yong Hwan;Oh, Sea-Kwan;Cho, Jun Hyeon;Song, You Chun
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.61 no.3
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    • pp.153-162
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    • 2016
  • We investigated the physicochemical properties and starch structure of various rice varieties including 15 colored cultivars. NKHC showed the highest level of protein, lipid, and total dietary fiber levels. Reddish brown rice showed higher lipid content than that in black rice cultivars. Apparent amylose content of waxy and non-waxy colored rice varieties was within the range of 3~5% and 15~18%, respectively. IP exhibited the highest total starch (TS) content, whereas, NKHC and HJJ showed lower TS content than that in other cultivars. Pasting temperature of all colored rice cultivars, except IP, was about $68^{\circ}C$. Peak viscosity of IP, JJJ, Hong, and GGHM showed high values of 138, 130, 128, and 124, respectively. All the colored rice cultivars presented A-type X-ray diffraction pattern and polygonal shapes of starch granules were observed using scanning electron micrographs (SEM). Major groups of amylopectin chain lengths were B (12 < DP ${\leq}$ 24) and A (DP ${\leq}$ 12). SMHC showed the highest B chain content and the lowest A chain content (P < 0.05). These experimental results provided useful information for scientists and the food industry regarding colored rice starches.