• Title/Summary/Keyword: collagen content

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Fixation and Histochemistry of Biological Tissues Using the Microwave Fixator Equipped with Infrared-Temperature Sensor (적외선 온도감응기를 장착한 마이크로파 고정기에 의한 생체조직 고정효과와 조직화학적 특성)

  • 신길상;민소연;김완종;손태호
    • The Korean Journal of Zoology
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    • v.38 no.3
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    • pp.417-425
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    • 1995
  • The present study was carried out to investigate the effect of microwave fixation in comparison with that of chemical fixation in preparing the microscopic samples. The microwave fixator was equipped with infrared-temperature sensor, and that was designed to compensate air temperature in the microwave fixator. In the microwave fixation, rat tongue was well preserved in terms of muscular fasciculus and pancreas stained by Feulgen reagents showed clear reaction products in the nucleus. Reaction products by PAS method in duodenal villi appeared specifically at the goblet cells. In electron microscopy, pancreatic cellular components such as secretory granules and collagen bundles were well preserved in both fixations. In aspect of histochemical reaction and electron microscopy, high quality was due to the protein content of microwave fixed specimen. The microwave fixation method saved total duration engaging microscopic preparation.

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The Anti-Fibrogenic Effect of a Pharmaceutical Composition of[5-(2-Pyrazinyl)-4-methyl-1,2-dithiol-3-thione] (Oltipraz) and Dimethyl-4,4′-dimethoxy-5,6,5′,6′-dimethylene dioxybiphenyl-2,2′-dicarboxylate (DDB)

  • Kang, Keon-Wook;Kim, Yoon-Gyoon;Kim, Choon-Won;Kim, Sang-Geon
    • Archives of Pharmacal Research
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    • v.25 no.5
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    • pp.655-663
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    • 2002
  • Liver fibrosis is a prepathological state wherein damaged liver tissues in chronic liver diseases, such as hepatitis, are not repaired to normal tissues, but converted to fibrous tissue. 5-(2-Pyrazinyl)-4-methyl-1,2-dithiol-3-thione (oltipraz), a cancer chemopreventive agent, is effective against a wide variety of chemical carcinogens. Recently, we reported that oltipraz inhibits liver fibrogenesis (Kang et al., 2002). In the present study, the effects of oltipraz in combination with dimethyl-4,4'-dimethoxy-5,6,5',6'-dimethylene dioxybiphenyl-2,2'-dicarboxylate (DDb) on dimethylnitrosamine (DMN)-induced liver fibrogenesis were assessed in rats. Oltipraz (30 mg/kg body weight, po, 3 times per week for 4 weeks) was found to inhibit the increases in plasma ALT, AST and bilirubin by DMN, whereas DDB (30 mg/kg body weight, po, 3 times per week for 4 weeks) attenuated the increases in the plasma ALT and bilirubin. The lowered plasma protein and albumin contents in DMN-treated rats were completely restored by oltipraz, but not by DDB. DDB decreases liver cell injury and inflammation through inhibition of nuclear factor-kB. DMN increased the accumulation of liver collagen, as indicated by the increase in the 4-hydroxyproline content in liver homogenates, which was reduced by treatment with oltipraz, but not by DDB. Given the differential effect between oltipraz and DDB, the potential enhancement of antifibrotic efficacy by the drugs was assessed in the animal model. Despite the minimal effect of DDB on DMN-induced fibrogenesis, DDB (5-25 mg/kg), administered together with oltipraz (25-5 mg/kg), showed an additive protective effect against hepatotoxicity and fibrosis induced by DMN, which was shown by the blood chemistry parameters and histopathological analysis. The adequate composition ratio of oltipraz to DDB was 5:1. These results provide information on the pharmaceutical composition, comprising of oltipraz and DDB as the active components, for the treatment and/or prevention of liver fibrosis and cirrhosis.

Metabolic engineering of Vit C: Biofortification of potato

  • Upadhyaya, Chandrama P.;Park, Se-Won
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2010.10a
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    • pp.14-14
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    • 2010
  • Vitamin C (ascorbic acid) is an essential component for collagen biosynthesis and also for the proper functioning of the cardiovascular system in humans. Unlike most of the animals, humans lack the ability to synthesize ascorbic acid on their own due to a mutation in the gene encoding the last enzyme of ascorbate biosynthesis. As a result, vitamin C must be obtained from dietary sources like plants. In this study, we have developed two different kinds of transgenic potato plants (Solanumtuberosum L. cv. Taedong Valley) overexpressing strawberry GalUR and mouse GLoase gene under the control of CaMV 35S promoter with increased ascorbic acid levels. Integration of the these genes in the plant genome was confirmed by PCR and Southern blotting. Ascorbic acid(AsA) levels in transgenic tubers were determined by high-performance liquid chromatography(HPLC). The over-expression of these genes resulted in 2-4 folds increase in AsA intransgenic potato and the levels of AsA were positively correlated with increased geneactivity. The transgenic lines with enhanced vitamin C content showed enhanced tolerance to abiotic stresses induced by methyl viologen(MV), NaCl or mannitol as compared to untransformed control plants. The leaf disc senescence assay showed better tolerance in transgenic lines by retaining higher chlorophyll as compared to the untransformed control plants. Present study demonstrated that the over-expression of these gene enhanced the level of AsA in potato tubers and these transgenics performed better under different abiotic stresses as compared to untransformed control. We have also investigated the mechanism of the abiotic stress tolerance upon enhancing the level of the ascorbate in transgenic potato. The transgenic potato plants overexpressing GalUR gene with enhanced accumulation of ascorbate were investigated to analyze the antioxidants activity of enzymes involved in the ascorbate-glutathione cycle and their tolerance mechanism against different abiotic stresses under invitro conditions. Transformed potato tubers subjected to various abiotic stresses induced by methyl viologen, sodium chloride and zinc chloride showed significant increase in the activities of superoxide dismutase(SOD, EC 1.15.1.1), catalase, enzymes of ascorbate-glutathione cycle enzymes such as ascorbate peroxidase(APX, EC 1.11.1.11), dehydroascorbate reductase(DHAR, EC 1.8.5.1), and glutathione reductase(GR, EC 1.8.1.7) as well as the levels of ascorbate, GSH and proline when compared to the untransformed tubers. The increased enzyme activities correlated with their mRNA transcript accumulation in the stressed transgenic tubers. Pronounced differences in redox status were also observed in stressed transgenic potato tubers that showed more tolerance to abiotic stresses when compared to untransformed tubers. From the present study, it is evident that improved to lerance against abiotic stresses in transgenic tubers is due to the increased activity of enzymes involved in the antioxidant system together with enhanced ascorbate accumulated in transformed tubers when compared to untransformed tubers. At moment we also investigating the role of enhanced reduced glutathione level for the maintenance of the methylglyoxal level as it is evident that methylglyoxal is a potent cytotoxic compound produced under the abiotic stress and the maintenance of the methylglyoxal level is important to survive the plant under stress conditions.

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Effects of feed intake restriction during late pregnancy on the function, anti-oxidation capability and acute phase protein synthesis of ovine liver

  • Yang, Huan;Wang, Ying;Ma, Chi;Sun, Chuan;Liu, Yingchun;Wu, Kaifeng;Li, Ming;Borjigin, Gerelt;Gao, Feng
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.2
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    • pp.217-223
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    • 2019
  • Objective: An experiment was conducted to investigate the effects of feed intake restriction during late pregnancy on the function, anti-oxidation capability and acute phase protein synthesis of ovine liver. Methods: Eighteen time-mated ewes with singleton fetuses were allocated to three groups: restricted group 1 (RG1, 0.18 MJ ME/kg $W^{0.75}$ d, n = 6), restricted group 2 (RG2, 0.33 MJ ME/kg $W^{0.75}$ d), n = 6) and a control group (CG, ad libitum, 0.67 MJ ME/kg $W^{0.75}$ d, n = 6). The feed restriction period was from 90 days to 140 days of pregnancy. Results: The ewe's body weight, liver weights, water, and protein content of liver in the restricted groups were reduced compared with the CG group (p<0.05), but the liver fat contents in the RG1 group were higher than those of the CG group (p<0.05). The increased hepatic collagen fibers and reticular fibers were observed in the restricted groups with the reduction of energy intake. The concentrations of nonesterified free fatty acids in the RG1 and RG2 groups were higher than those of the CG group with the reduction of energy intake (p<0.05), but there were decreased concentrations of lipoprotein lipase and hepatic lipase in both restricted groups compared with the CG group (p<0.05). In addition, the increased concentrations of ${\beta}$-hydroxybutyric acid, triglycerides, malondialdehyde, total antioxidant capacity and activities of superoxide dismutase activity and catalase were found in the RG1 group, and the concentrations of cholinesterase in the RG1 group were reduced compared with the CG group (p<0.05). For the concentrations of acute phase proteins, the C-reactive protein (CRP) in the RG1 group were reduced compared with the CG group, but there were no differences in haptoglobin relative to the controls (p>0.05). Conclusion: The fat accumulation, increased hepatic fibrosis, antioxidant imbalance and modified synthesis of acute phase proteins were induced in ewe's liver by maternal malnutrition during late pregnancy, which were detrimental for liver function to accommodate pregnancy.

Acidification of drinking water improved tibia mass of broilers through the alterations of intestinal barrier and microbiota

  • Zhang, Huaiyong;Guo, Yujun;Wang, Ziyang;Wang, Yongshuai;Chen, Bo;Du, Pengfei;Zhang, Xiangli;Huang, Yanqun;Li, Peng;Michiels, Joris;Chen, Wen
    • Animal Bioscience
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    • v.35 no.6
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    • pp.902-915
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    • 2022
  • Objective: Diet acidification supplementation is known to influence intestinal morphology, gut microbiota, and on phosphorus (P) utilization of broilers. Alterations in intestinal barrier and microbiota have been associated with systemic inflammation and thus regulating bone turnover. Hence the effect of acidifier addition to drinking water on tibia mass and the linkages between intestinal integrity and bone were studied. Methods: One-d-old male broilers were randomly assigned to normal water (control) or continuous supply of acidified water (2% the blend of 2-hydroxy-4-methylthiobutyric acid, lactic, and phosphoric acid) group with 5 replicates of 10 chicks per replicate for 42 d. Results: Acidification of drinking water improved the ash percentage and calcium content of tibia at 42 d. Broilers receiving acidified water had increased serum P concentration compared to control birds. The acidified group showed improved intestinal barrier, evidenced by increased wall thickness, villus height, the villus height to crypt depth ratio, and upregulated mucin-2 expression in ileum. Broilers receiving drinking water containing mixed organic acids had a higher proportion of Firmicutes and the ratio of Firmicutes and Bacteroidetes, as well as a lower population of Proteobacteria. Meanwhile, the addition of acidifier to drinking water resulted in declined ileal and serum proinflammatory factors level and increased immunoglobulin concentrations in serum. Concerning bone remodeling, acidifier addition was linked to a decrease in serum C-terminal cross-linked telopeptide of type I collagen and tartrate-resistant acid phosphatase reflecting bone resorption, whereas it did not apparently change serum alkaline phosphatase activity that is a bone formation marker. Conclusion: Acidified drinking water increased tibia mineral deposition of broilers, which was probably linked with higher P utilization and decreased bone resorption through improved intestinal integrity and gut microbiota and through decreased systemic inflammation.

Effect of Cryoprotectants on the Physico-chemical Characteristics of Chicken Breast Surimi Manufactured by pH Adjustment during Freezing Storage (냉동변성 방지제가 pH 조절법으로 제조한 닭가슴살 수리미의 냉동저장 중 이화학적 특성에 미치는 영향)

  • Jin, Sang-Keun;Kim, Il-Suk;Kim, Su-Jung;Jeong, Ki-Jong;Lee, Jae-Ryong;Choi, Yeung-Joon
    • Food Science of Animal Resources
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    • v.27 no.3
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    • pp.267-276
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    • 2007
  • This study was conducted to determine the effect of cryoprotectants (sugar, sorbitol, polyphosphate) on the physico-chemical characteristics of chicken breast surimi manufactured by pH adjustment (pH 11.0) during freezing storage. The final surimi was divided into experimental units to which the following treatments were randomly assigned: C (Alaska pollack surimi: two washings, 4% sugar +5% sorbitol ${\pounds}'$ 0.3% polyphosphate additive): T1 (chicken breast surimi: pH 11.0 adjusted, 0.3% polyphosphate additive): T2 (chicken breast surimi pH 11.0 adjusted, 5% sorbitol +0.3% polyphosphate additive); T3 (chicken breast surimi: pH 11.0 adjusted, 4% sugar +5% sorbitol +0.3% polyphosphate additive). The crude protein content of the control was higher than all treated samples, however the moisture, crude fat and crude ash of T3 were higher than the control (p<0.05). The pH, WHC and collagen content of the control were higher than all of the treated samples, and these values decreased with storage time for all treatments and the control (p<0.05). The cholesterol content of the control was lower than all treated samples, but the myofibrillar protein contents of all treated samples were higher than the control (p<0.05). The cooking loss of T2 was lower than the control and the other two treatments (p<0.05). The $L^*,\;a^*\;and\;b^*$ values of all treated samples were higher than those of the control during freezing storage (p<0.05). The W value of T3 at 1.5 and 3 months of freezing storage was higher than the control and T1 (p<0.05). The myoglobin and met-Mb contents of the control were similar to all treated samples, and the met-Mb content of the control and all treated samples increased with storage time (p<0.05). Immediately after freezing, the hardness of the control was higher than all treated samples, however it was lower after 1.5 and 3 months of frozen storage (p<0.05). The cohesiveness and gumminess of the control were higher than all treated samples immediately after freezing, however the values for T3 were higher than those of the control and the other two treatments during frozen storage for 1.5 and 3 months (p<0.05).

Effect of 2-D DBP/PLGA Hybrid Films on Attachment and Proliferation of Intervertebral Disc Cells (2차원적 DBP/PLGA 하이브리드 필름이 디스크 세포의 부착과 증식에 미치는 영향)

  • Ko, Youn-Kyung;Jeong, Jae-Soo;Kim, Soon-Hee;Lim, Ji-Ye;Rhee, John-M.;Kim, Moon-Suk;Lee, Hai-Bang;Khang, Gil-Son
    • Polymer(Korea)
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    • v.32 no.2
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    • pp.109-115
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    • 2008
  • Because demineralized bone particle (DBP) contains various bioactive molecules such as cytokines, it is widely used biomaterials in the field of tissue engineering. In this study, we investigated the effect of 2-dimensional DBP/PLGA hybrid films on adhesion, proliferation and phenotype maintenance of intervertebral disc cells. PLGA films incorporated with different amount (0, 10, 20, 40 and 80 wt%) of DBP were prepared by the solvent evaporation method and characterized by scanning election microscopy (SEM). PLGA film has a flat and smooth surface. According to the increase of content of DBP, the surface of DBP/PLGA film exhibited few agglomerates and increased the roughness of the surface. Annulus fibrosus (AF) and nucleus pulposus (NP) cells were cultured on PLGA and DBP/PLGA film surface, and then examined the cell adhesion and proliferation by the cell count and SEM observation. The result of cell count and SEM observation revealed that 10 and 20% DBP in DBP/PLGA films were superior to adhesion and proliferation of both AF and NP cells. We confirmed that specific gene expression of disc cells on DBP/PLGA film based on the cell count result. Disc cells seeded on 20% DBP/PLGA film expressed the gene of type I and II collagen continuously. Therefore, pertinent content of biomaterials could provide more appropriate condition on adhesion and proliferation of cell. And this results may be used as a basic data for the intervertebral disc regeneration using tissue engineering.

Processing and Quality Properties of Olive Flounder Paralichthys olivaceus Cutlet (넙치(Paralichthys olivaceus) 커틀렛 제품의 제조 및 품질특성)

  • YOON, Moon-Joo;LEE, Jae-Dong;KWON, Soon-Jae;PARK, Si-Young;KONG, Cheong-Sik;JOO, Jong-Chan;KIM, Jeong-Gyun
    • Journal of Fisheries and Marine Sciences Education
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    • v.27 no.3
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    • pp.625-633
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    • 2015
  • Olive flounder (Parlichthys olivaceus) is a large carnivorous fish that live at coastal area and shallow seas in Korea. It was good texture and clean taste because of a high collagen content and low lipid content. More than 70% of olive flounder annual production was traded alive, consequently processing food product from olive flounder is rare to be towed. This study was conducted to investigate the best method of olive flounder cutlet processing. Clean fillet (headless, skinless and contain no viscera part) of olive flounder were divided into 5 portion. Every 100 g of olive flounder meat was wrapped with vinyl then flatten with meat hammer. Flatten fillet then was coated with wheat flour, and seasoned with salt and pepper. These were then coated with egg wash and bread crumbs. Two different method of processing was to make this olive flounder cutlet. Cutlet-1 was fried for 1 min in olive oil, then kept in polyethylene film vacuum packaging ($20{\times}30{\times}0.05mm$) and stored at $-20^{\circ}C$ for 7 days. After 7 days the cutlet was thawed and heat up in microwave for 2 min (Sample-1). The other proup is cutlet-2, which is directly stored in polyethylene film vacuum packaging at $-20^{\circ}C$ for 7 days then thawed and fried for 1 min in olive oil (Sample-2). The factors such as pH, TBA value, amino-N, free amino acid, chemical composition, color value (L, a, b), texture profile, sensory evaluation and viable bacterial count of the olive flounder cutlet (Sample-1, Sample-2) were measured. From the result of sensory evaluation, Sample-2 showed a little high scores than Sample-1. But there was no significant differences in color, odor, taste, texture and overall acceptance between Sample-1 and Sample-2 products.

Effect of Ratio of Demineralized Bone Powder with Alginate Microcapsules on Articular Cartilage Regeneration (탈미네랄 골분이 비율별로 포접된 알지네이트 미세캡슐을 이용한 조직공학적 연골재생)

  • Kim, A Ram;Kim, Hye Min;Lee, Jung Keun;Lee, Ji Hye;Song, Jeong Eun;Yoon, Kun Ho;Lee, Dongwon;Khang, Gilson
    • Polymer(Korea)
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    • v.36 no.6
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    • pp.768-775
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    • 2012
  • Alginate, obtained from the seaweeds, is a widely used biomaterial for cell transplantation, since its positive effect on viability of capsulized cells and its easier encapsulation capability of living cells. Demineralized bone powder (DBP), derived from the natural bone tissue, is widely applied for clinical trials for its low rate of reaction and antigenicity. A chondrocyte was seeded into an alginate with DBP of different contents, and a microcapsule was produced. The adhesion and proliferation of cells was observed through the MTT analysis, and the PCR was applied to estimate the content of the glycosaminoglycan (sGAG) and collagen, and confirm the specific genetic pattern of the chondrocytes. Also, the alginate microcapsule where the chondrocyte is seeded was extracted after transplantation under the skin of a nude mouse, and was immunochemically stained. The experimental result confirmed that the alginate microcapsule containing 1% of DBP not only showed the highest proliferation of cell but had a positive effect of chondrocytes by the interaction between the alginates and the growth factor in DBP. It can be expected that the microcapsule with application of the alginates and DBP might be an appropriate scaffold for tissue engineering.

Skin Protection Effect of Grape Pruning Stem Extract on UVB-induced Connective Tissue Injury (포도전정가지 추출물이 UVB로 유도된 결합 조직 손상에 미치는 피부 보호 효과)

  • Kim, Joung-Hee;Kim, Keuk-Jun
    • Journal of Life Science
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    • v.28 no.2
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    • pp.141-147
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    • 2018
  • This study aimed to analyze the contents of rutin, procyanidin B3, quercetin, and kaempferol, known to have antioxidant, anti-inflammatory, and anti-carcinogenic effects, among the polyphenol types contained in grape pruning stem extracts (GPSE). It utilized grape stems discarded after harvest to measure the effects of GPSE on skin moisture, inhibition of skin cell proliferation, and anti-inflammatory activity on the damaged skin of HR-1 mice induced with ultraviolet B (UVB), and to verify the applicability of GPSE as a material for functional food and functional cosmetics. The polyphenol was extracted from grape pruning stems with 80% EtOH, and then the extract was used while storing at $-20^{\circ}C$, after filtering, concentrating, and freeze-drying it. The content of an active ingredient of GPSE was analyzed using high performance liquid chromatography (HPLC). From 53 kg of the grape pruning stem specimen, 2.34 kg of the EtOH fraction extracts were extracted to achieve a 4.42% yield ratio. Analysis of the active ingredients showed 0.28 mg/g of procyanidin B3, 12.81 mg/g of rutin, 0.51 mg/g of quercetin, and 8.24 mg/g of kaempferol. After UVB irradiation on the dermis, to confirm the degree of inhibition of collagen synthesis, we examined the protein expression of MMP-9 using immunohistochemical staining. The results of this study confirm the existence of active polyphenol types, such as rutin, kaempferol, quercetin, and procyanidin B3, in GPSE. Moreover, the study found that GPSE has anti-collagenase effects and it decreases the effects of UV damage on skin barrier function. GPSE is a functional ingredient with a potential for skin protection effects, and it has high utilization potential as an ingredient for functional cosmetics.