• Title/Summary/Keyword: Incubation period

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STIMULATION OF OSTEOBLASTIC PHENOTYPES BY STRONTIUM IN PERIOSTEAL-DERIVED CELLS (골막기원세포에서 strontium에 의한 조골세포 표현형의 활성)

  • Kim, Shin-Won;Kim, Uk-Kyu;Park, Bong-Wook;Hah, Young-Sool;Cho, Hee-Young;Kim, Jung-Hwan;Kim, Deok-Ryong;Kim, Jong-Ryoul;Joo, Hyun-Ho;Byun, June-Ho
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.32 no.3
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    • pp.199-206
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    • 2010
  • This study investigated the effects of strontium on osteoblastic phenotypes of cultured human periostealderived cells. Periosteal tissues were harvested from mandible during surgical extraction of lower impacted third molar. Periosteal-derived cells were introduced into cell culture. After passage 3, the periostealderived cells were further cultured for 28 days in an osteogenic induction DMEM medium supplemented with fetal bovine serum, ascorbic acid 2-phosphate, dexamethasone and at a density of $3{\times}10^4$ cells/well in a 6-well plate. In this culture medium, strontium at different concentrations (1, 5, 10, and 100 ${\mu}g$/mL) was added. The medium was changed every 3 days during the incubation period. We examined the cellular proliferation, histochemical detection and biochemical measurements of alkaline phosphatase (ALP), the RT-PCR analysis for ALP and osteocalcin, and von Kossa staining and calcium contents in the periostealderived cells. Cell proliferation was not associated with the addition of strontium in periosteal-derived cells. The ALP activity in the periosteal-derived cells was higher in 5, 10, and 100 ${\mu}g$/ml strontium-treated cells than in untreated cells at day 14 of culture. Among the strontium-treated cells, the ALP activity was appreciably higher in 100 ${\mu}g$/ml strontium-treated cells than in 5 and 10 ${\mu}g$/ml strontium-treated cells. The levels of ALP and osteocalcin mRNA in the periosteal-derived cells was also higher in strontium-treated cells than in untreated cells at day 14 of culture. Their levels were increased in a dose-dependent manner. Von Kossa-positive mineralization nodules were strongly observed in the 1 ${\mu}g$/ml strontium-treated cells at day 21 and 28 of culture. The calcium content in the periosteal-derived cells was also higher in 1 ${\mu}g$/ml strontium-treated cells at day 28 of culture. These results suggest that low concentration of strontium stimulates the osteoblastic phenotypes of more differentiated periosteal-derived cells, whereas high concentration of strontium stimulates the osteoblastic phenotypes of less differentiated periosteal-derived cells. The effects of strontium on osteoblastic phenotypes of periosteal-derived cells appear to be associated with differentiation-extent.

Effects of Caffeine and calcium on the activities of the mouse osteoblastic cells (카페인과 칼슘이 골모 세포의 활성에 미치는 영향)

  • Chun, Youn-Sic;Baik, Hye-Jung
    • The korean journal of orthodontics
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    • v.32 no.2 s.91
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    • pp.129-142
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    • 2002
  • The purpose of this study was to evaluate the effects of caffeine and calcium on the activities of the osteoblastic cell from mouse calvaria. The author cultured osteoblastic cells obtained from the mouse calvaria and were divided into three groups : the caffeine-treated, the calcium-treated and the combine-treated group. In caffeine-treated group, the cell toxicity was measured by MTT assay at 1, 2 and 4 days after treatment of caffeine. In all groups, the densities of the mineralized bone nodules were measured by imaging analyzer after Von Kossa staining. The alkaline phosphotase (ALP) activities were measured at 2, 7, 14, 21 and 28 days and the interleukin-1 ${\beta}$ activities at 48 hours after treatment of caffeine and calcium. The measurements were statistically executed with ANOVA test and the results were as follows. 1. The cellular toxicity of the caffeine increased with the concentration of caffeine during the incubation period. 2. The maximum densities of mineralization were observed at 0.2 mM caffeine-treated group, 1.2 mM calcium-treated group, 0.1 mM caffeine and 1.8 mM calcium-treated group. 3. The activities of ALP were peaked at 14 days at calcium-treated group as no-treated. But, the activities of ALP increased with concentrations of caffeine at caffeine-treated group. At combine-treated group, the act of ALP were peaked at 24 days at 1.2 mM, 1.8 mM calcium-treated group, But decreased at 2.5 mM calcium-treated group. 4. The activites of the IL-1 ${\beta}$ were increased significantly at 0.2 mM caffeine-treated group, 1.8 mM calcium-treated group and 0.1 mM caffeine and 1.8 mM calcium-treated group. But, they were decreased at all groups of high concentration.

Antimicrobial and Anticancer Activity of Korean Traditional Soy Sauce and Paste with Chopi (초피첨가 전통장류의 항균 및 항암활성)

  • Kim, Keun-Ki;Park, Hyean-Cheal;Son, Hong-Joo;Kim, Yong-Gyun;Lee, Sang-Mong;Choi, In-Soo;Choi, Young-Whan;Shin, Teak-Soon
    • Journal of Life Science
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    • v.17 no.8 s.88
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    • pp.1121-1128
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    • 2007
  • The fruits of Zanthoxylum piperitum are known as having various physiology vitality, and the abstraction ingredient of the pericarp is also known as having strong antibiotic activities against various bacteria. Therefore, this study was carried out to estimate the effect of physiology vitality when the abstraction ingredient of Z. piperitum was added in soy sauce(Chopi-kanjang) and soybean paste(Chopi-doenjang). For the antibiotic activity against the pathogens of sitotoxism such as Staphylococcus aureus, Salmonella typhimurium, Vibrio parahemolyticus, Escherichia coli 0157:H7, the extracts of the Chopi-kanjang was added 1%, 2%, 4% pericarp of Z. piperitum in the manufacturing process of soy source. According to the results, the growth of E. coli 0157:H7 and V. parahemolyticus were respectively inhibited as 70% and 50% by the Chopi-kanjang added 2% of the ingredient. For the antibiotic effects of the aforementioned Chopi-kanjang against Sal. typhimurium and Sta. aureus, the growth of those pathogens was also inhibited between 40% and 60% according to the manufacturing period of Chopi-kanjang. It was confirmed that the antibiotic activity using the mixture of the abstraction ingredient and Chopi-doenjang was lower than those of Chopi-kanjang. In order to estimate the anticancer activity using by caspase-3 activity, the mixture of the abstraction ingredient of the pericarp of Z. piperitum and Chopi-kanjang was treated to leukemia cells. According to the results, the activities of caspase-3 using the mixture added 1%, 2% and 4% of the abstraction ingredient were respectively increased as much as 4, 12, 15 times comparing with the control which was treated with the soy source only. It could be that the mixture of the abstraction ingredient of the pericarp of Z. piperitum and soy source induced apoptosis, and the mixture of the abstraction ingredient and soybean paste had no effect on the activity of caspase-3. In order to find out the death of the aforementioned cells caused by necrosis or apoptosis, DNA fragmentation in the cell was examined. U-937 cells showed apoptotic DNA fragmentation in the incubation with Chopi-kanjang extract.

Effects of Protein and Carbohydrate Supplements on Feed Digestion in Indigenous Malaysian Goats and Sheep

  • Darlis, N. Abdullah;Halim, R.A.;Jalaludin, S.;Ho, Y.W.
    • Asian-Australasian Journal of Animal Sciences
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    • v.13 no.4
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    • pp.464-469
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    • 2000
  • Experiments were conducted to determine the effects of soybean meal (SBM) as a source of protein and sago meal (SM) as a source of carbohydrate on in situ and in vivo digestibility of dietary components in four male goats (Kambing Katjang) and four male sheep (Malin) weighing 25-35 kg. Rumen volume, as well as rumen fluid dilution rate were also determined. The animals were housed in single pens with individual feeding and drinking troughs and each animal was fitted with a rumen fistula. They were fed two diets : chopped rice straw+200 g soybean meal (SBM), and chopped rice straw+190 g soybean meal+300 g sago meal (SBM+SM). Rice straw was offered ad libitum. The supplements were isonitrogenous (80 g crude protein/animal/d), but the proportions of dry matter (DM), organic matter (OM), crude fibre (CF), neutral detergent fibre (NDF) and acid detergent fibre (ADF) were lower in the SBM supplement (191, 165, 11, 40, 15 g/animal/d for DM, OM, CF, NDF and ADF, respectively) than in the SBM+SM supplement (445, 423, 25, 102, 38 g/animal/d for DM, OM, CF, NDF and ADF, respectively). Two animals from each species were fed either supplement in a cross-over design in two periods. Each period lasted for four weeks. In situ and in vivo digestibility studies were carried out, followed by the determination of rumen volume and rumen fluid dilution rate. The results showed that straw DM and total DM intakes of goats (average of $48.7g/kg\;W^{0.75}$, $72.7g/kg\;W^{0.75}$, respectively) were significantly (p<0.01) higher than sheep (average of $3.56g/kg\;W^{0.75}$, $61.6g/kg\;W^{0.75}$, respectively), but OM, N and GE intakes were not significantly different between the two animal species. When the effect of supplements was compared, animals fed SBM+SM supplement had significantly (p<0.001) higher DM, OM and GE intakes than animals fed SBM supplement. Potential degradabilities of rice straw DM were significantly (p<0.01) higher in goats (average of 48.8%) than in sheep (average of 46.1 %). The supplements had no significant effect on the potential degradabilities of DM, OM and NDF, but they had a significant (p<0.05) effect on the degradation rates of DM and NDF. The addition of sago meal in the diet reduced the degradation rates of DM and NDF of rice straw in the rumen. Potential degradability of DM of soybean meal was not significantly different between animal species as well as between supplements. Sago meal was highly degradable. At 24 h of incubation in the rumen, 90-95% of DM loss was observed. There was a significant interaction between animal species and supplements in the in vivo digestibility of ADF and GE. In animals fed SBM supplement, the in vivo digestibility of ADF was significantly (p<0.05) higher in goats ($50.6{\pm}4.22%$) than in sheep ($44.4{\pm}3.21%$), but digestibility of GE was significantly (p<0.05) higher in sheep ($70.2{\pm}1.93%$) than in goats ($63.0{\pm}3.07%$). The digestibility values of CP and OM were significantly (p<0.05) higher in sheep when compared to goats. Animals fed SBM+SM supplement showed significantly (p<0.05) higher DM and OM digestibility values than animals fed SBM supplement, but digestibility values of CP were significantly (p<0.05) higher in animals fed SBM supplement. Differences in in vivo digestibility values of CF and NDF were not significantly different between animal species or supplements. Water intake, rumen volume ($1/kg\;W^{0.75}$), rumen fluid dilution rate and mean retention time were similar between the two animal species. However, rumen fluid dilution rate and mean retention time was significantly (p<0.01) affected by supplements. Animals fed SBM+SM had faster rumen fluid dilution rate and consequently shorter mean retention time.

Reproductive Progress and Heavy Metal Contamination of Feral Pigeons (Columba livia) in Seoul and the Ansan Industrial Complex areas (도심 및 공단지역에 서식하는 비둘기의 중금속 축적과 번식 경과 비교)

  • 남동하;이두표;구태회
    • Korean Journal of Environmental Biology
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    • v.21 no.2
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    • pp.149-157
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    • 2003
  • This study focused on relationships between Pb and Cd concentrations and the difference of success reproductive progress in urban (Seoul) and industrial complex (Ansan) areas. Results of the Pb analysis for the feral pigeons from Seoul (egg contents:1.64 $\mu\textrm{g}$ wet $g^{-1}$, adults in bones: 29.5 $\mu\textrm{g}$ wet $g^{-1}$ and the Ansan industrial complex (egg contents: 1.13 $\mu\textrm{g}$ wet $g^{-1}$, adults in bones: 10.5 $\mu\textrm{g}$ wet g-1) showed that the Pb level of eggs and adults is significantly different between the two colonies (p<0.05). Cd concentrations in liver and kidneys of adult pigeons were also significantly different between Seoul(liver: 0.24 ${\mu}g$ wet $g^{-1}$, kidney: 1.05 $\mu\textrm{g}$ wet $g^{-1}$ and the Ansan (liver: 0.14 $\mu\textrm{g}$ wet $g^{-1}$, kidney: 0.43 $\mu\textrm{g}$ wet $g^{-1}$ colonies (P<0.05). Clutch size of Feral Pigeons living in Seoul was similar between the two colonies, 1.9$\pm$0.3 in Seoul and 2.0$\pm$0.0 in Ansan. The length, breadth, and thickness of eggs were not significantly different between the two colonies (p>0.05). Incubation period in Seoul (17.8 days) did not differ from the Ansan (17.4 days). No difference in growth rate (body weight, wing length, and tarsus length) was found between the two test groups (p>0.05). In Seoul, 65.2% were hatching, and 42.1% fledging. The Proportion of hatching and fledging in the Ansan was 60.7% and 45.0%, respectively. The significant differences between the two colonies for reproductive sucess were not found (p>0.05). With regard to the reproductive effects to the heavy metals, the Pb and Cd concentrations feund in the two colonies were not as high as those considered in results of toxic effects in other species.

Effect of Media, Synchronization of Fibroblast Cells, Culture Time, $\textrm{O}_2$ Concentration and Activation on Developmental Rate of Nuclear Transferred Porcine Oocytes (배양액, 섬유아세포, 배양시간, 산소 농도 및 활성화 처리가 돼지 핵이식 배의 체외발달에 미치는 영향)

  • Quan J. H.;Rhee M. H.;Kim S. K.
    • Journal of Embryo Transfer
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    • v.19 no.3
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    • pp.191-199
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    • 2004
  • This study was conducted to examine in vitro development of porcine embryos constructed by the microinjection of cultured fetal fibroblast cells into porcine oocytes matured in vitro. Single fetal donor cells were deposited into the perivitelline space of enucleated oocytes, followed by electrical fusion and activation. Activated embryos were cultured in NCSU-23 medium supplemented with 5% FBS, at 38.5$^{\circ}C$ for 6 to 8 days in 5% $CO_2$ and air. In experiment 1, fusion rates of nuclear transfer embryos did not differ for fetal fibroblast cells incubated in 5% FBS + NCSU-23 or 5% FBS + TL Heaps medium, nor did fusion rates of donor cells differ between 1-8 hr incubation durations. Fusion rates for the four treatment subclasses ranged from 72.1% to 78.0%. In experiment 2, Pre-synchronization in medium containing 0.1 $\mu\textrm{g}$/m Hoechst 33342 an increase from 0 and 8 versus 15 h culture an increased percentage of porcine fibroblast cells in G2/M at the end of the synchronization period (12.4%, 17.5% and 47.6%). Neither an increase in the concentration of H 33342 (0.2-1.6 $\mu\textrm{g}$/$m\ell$) nor a longer exposure time (12h, 18h and 24h) increased the proportion of porcine G2/M fibroblasts. In experiment 3, fusion rates did not differ significantly far nuclear transfer embryos constructed using donor cells cultured in 5% FBS + NCSU-23 medium for 1-2, 6-8 or 12-14 days (60.0%, 73.3% and 62.5%), respectively. The cleavage rate for nuclear transplant embryos using fetal fibroblast cells cultured for 1-2 days was 44.0%, significantly less than 56.7% and 50.0%. for 6-8 or 12-14 days duration of culture, respectively. In experiment 4, the proportions of nuclear transfer embryos that developed to the $\geq$2 cell and to the blastocyst stage were not affected significantly by culture medium (5% FBS + NCSU-23 or 5% FBS + TL-Heaps) or by $O_2$ concentration of the culture (5% vs 10%). Rates of development to the $\geq$2 cell stage ranged from 65.9% to 70.1%, and development rates to the blastocyst stage ranged from 9.8% to 12.5% for the four treatment subclasses. Developmental rate was highest for embryos cultured in 5% FBS + NCSU-23 under a gas atmosphere of 5% $O_2$ in air.

Studies on the Propagation of the Freshwater Prawn, Macrobrachium nipponense (De Haan) Reared in the Laboratory (담수산 징거미새우, Macrobrachium nipponense (De Haan)의 증${\cdot}$양식에 관한 생물학적 기초연구 1. 생식생태에 관한 연구)

  • Kwon Chin-Soo;Lee Bok-Kyu
    • Journal of Aquaculture
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    • v.4 no.1
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    • pp.31-66
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    • 1991
  • This paper deals with the reproductive ecology e.g., number of the pre-spawning moults, morphological characteristics of the pre-spawning moult the common moult, daily ration druing a molting cycle mating behavior, structures of spermatozoa and spermatophore, structure of vas deferens, mechanisms of the oviposition and brooding into the egg-chambers, a suitable time for the artificial mating and fertilization, time sequence of the oviposition and brooding into egg-chambers from the copulation, responses to temperature and chlorinity on the egg development and hatching, effect of temperatures on duration of egg development, physical mechanism of the egg hatching, to make an attempt for the artificial spawning and brooding to establish a suitable system of the artificial seedling-production for the aquaculture. 1. Females molted commonly $8{\~}10$ times at an interval of $17{\~}18$ days at $28^{\circ}C,\;3.26\~4.35\%_{\circ}$ while the prespawning moltings were $4{\~}5$ times at an interval of $13{\~}14$ days. The suitable state for artificial copulation was within 14 hours elapsed from the prespawning moltings (most suitable state was within 8 hours). Males discharged a gelatinous spermatophore and placed it on the females sternum during copulation. Oviposition was seen $6{\~}17$ hours after copulation. External fertilization was considered to take place at oviposition. Fertilized eggs held in egg-chambers forming between pleopods were about $5000{\~}6000$ in females those sizes about 6.5 cm in body length. 2. Eggs immediately after oviposition were elliptic shape, measuring $0.58{\times}0.48$ mm up to hatching. Their sizes increased with egg development and finally reached $0.85{\times}0.54$ mm up to hatching. The relationship between the long axis of the egg(Y in U) and days elapsed(X) was expressed as Y= 5.60194 + 0.007358X. The eggs performed superficial cleavage and their cleavage furrows became visible at the 4-daughter-nucleus stage. The eggs showed normal development up to hatching at water temperature range of $22{\~}30^{\circ}C$ (optimum temperature : $26{\~}28^{\circ}C$) and at chlorinity range of $0.00\~6.64\%_{\circ}$ (optimun chlorinity : $2.21{\%}_{\circ}$). The relationship between incubation period (Y in days) and water temperature(X in $^{\circ}C$) could be expressed as Y= 50.803-1.3555X. The eggs hatched $12{\~}13$ days after oviposition at $28.0{\~}28.6^{\circ}C$ 3. The pre-spawning moltings were appreciably different in the morphologic structure from those of common moltings. Breeding setae and dresses were formed on the thoracic regions, abdominal epimerae and the bases of the first to fourth pleopods in order to prepare and support oviposition, transfering and supporting eggs in egg-chambers up to hatching. These supplementary breeding organs were observed only at reproductive seasons.

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Selection for Duration of Fertility and Mule Duck White Plumage Colour in a Synthetic Strain of Ducks (Anas platyrhynchos)

  • Liu, H.C.;Huang, J.F.;Lee, S.R.;Liu, H.L.;Hsieh, C.H.;Huang, C.W.;Huang, M.C.;Tai, C.;Poivey, J.P.;Rouvier, R.;Cheng, Y.S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.28 no.5
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    • pp.605-611
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    • 2015
  • A synthetic strain of ducks (Anas platyrhynchos) was developed by introducing genes for long duration of fertility to be used as mother of mule ducklings and a seven-generation selection experiment was conducted to increase the number of fertile eggs after a single artificial insemination (AI) with pooled Muscovy semen. Reciprocal crossbreeding between Brown Tsaiya LRI-2 (with long duration of fertility) and Pekin L-201 (with white plumage mule ducklings) ducks produced the G0. Then G1 were intercrossed to produce G2 and so on for the following generations. Each female duck was inseminated 3 times, at 26, 29, and 32 weeks of age. The eggs were collected for 14 days from day 2 after AI. Individual data regarding the number of incubated eggs (Ie), the number of fertile eggs at candling at day 7 of incubation (F), the total number of dead embryos (M), the maximum duration of fertility (Dm) and the number of hatched mule ducklings (H) with plumage colour were recorded. The selection criterion was the breeding values of the best linear unbiased prediction animal model for F. The results show high percentage of exhibited heterosis in G2 for traits to improve (19.1% for F and 12.9% for H); F with a value of 5.92 (vs 3.74 in the Pekin L-201) was improved in the G2. Heritabilities were found to be low for Ie ($h^2=0.07{\pm}0.03$) and M ($h^2=0.07{\pm}0.01$), moderately low for Dm ($h^2=0.13{\pm}0.02$), of medium values for H ($h^2=0.20{\pm}0.03$) and F ($h^2=0.23{\pm}0.03$). High and favourable genetic correlations existed between F and Dm ($r_g=0.93$), between F and H ($r_g=0.97$) and between Dm and H ($r_g=0.90$). The selection experiment showed a positive trend for phenotypic values of F (6.38 fertile eggs in G10 of synthetic strain vs 5.59 eggs in G4, and 3.74 eggs in Pekin L-201), with correlated response for increasing H (5.73 ducklings in G10 vs 4.86 in G4, and 3.09 ducklings in Pekin L-201) and maximum duration of the fertile period without increasing the embryo mortality rate. The average predicted genetic response for F was 40% of genetic standard deviation per generation of selection. The mule ducklings' feather colour also was improved. It was concluded that this study provided results for a better understanding of the genetics of the duration of fertility traits in the common female duck bred for mule and that the selection of a synthetic strain was effective method of improvement.

Studies on Quality Changes and Antioxidant Activity During the Fermentation of the Salt Fermented Whangseoke (황석어(Collichthys nireatus Jordan et starks) 젓갈의 숙성과정 중 품질변화와 항산화작용에 관한 연구)

  • Kim, Ji-Sang;Moon, Gap-Soon;Lee, Kyung-Hee;Lee, Young-Soon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.35 no.2
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    • pp.171-176
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    • 2006
  • The effect of storage temperature on the quality and antioxidative activity Whangseoke sauce was studied over a period of 240 days. Fermented Whangseoke with $25\%$ salt were stored at $25^{\circ}C$. The quality change and antioxidant activity of Whangseoke in linoleic acid emulsion was evaluated with various parameters including acids values, peroxide values, TBA values, reducing sugar, brown color intensity, electron donating ability and reducing power at various time intervals for 240 days of storage. In general, it was observed, in all sample, that peroxide values, brown color intensity, electron donating ability and reducing power gradually increased, while reducing sugar decreased during storage at $25^{\circ}C$. The antioxidative activities of fermented Whangseoke were determined on tile linoleic acid emulsion system. The results showed that Whangseoke had antioxidant activity. These results suggest that antioxidant activity of Whangseoke seemed to influence by Maillard reaction products during the storage periods.

Effect of Boric Acid on In Vitro Pollen Germination in Transgenic Plants Expressing Monoclonal Antibodies (단일항체를 발현하는 형질전환 식물체의 In Vitro 화분발아에 대한 Boric Acid의 영향)

  • Ahn, Mi-Hyun;Lee, Kyung-Jin;Ko, Ki-Sung
    • Journal of Plant Biotechnology
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    • v.34 no.4
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    • pp.339-346
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    • 2007
  • Pollen germination viability is an essential factor to produce seeds from pollination and fertilization, which are required to maintain plant generation. In this study, we tried to identify the effect of boric acid on pollen germination and tube grouch in non-transgenic and transgenic plants expressing monoclonal antibodies (anti-colorectal cancer mAb CO17-1A, anti-breast cancer mAb BR55, and anti-rabies virus mAb57). The pollen of non-transgenic plant was treated with different concentration of boric acid (0, 5, 10, 15, 20, $40{\mu}g/mL$) in germination buffer to investigate its effect on in vitro pollen germination. At $20{\mu}g/mL$ of boric acid, tile pollen germination rate was the highest (49.5%) compared to other concentrations. In general, the germination rate significantly increased 3-10 folds in boric acid ($20{\mu}g/mL$) treated group in non-transgenic and transgenic plants. Also, the pollen tube length increased in boric acid ($20{\mu}g/mL$) treated groups. In the treated group, the pollen tube length increased until 3 h boric acid treatment and decreased after the 3 h, indicating that the 3 h is the most appropriate incubation time period. Western blot analysis showed that the mAb transgene expression was more stable in leaf than pollen in transgenic plants. This study suggested that $20{\mu}g/mL$ of boric acid is ideal concentration to induce in vitro pollen germination of transgenic plants expressing therapeutic monoclonal antibodies, indicating stable pollination and fertilization in transgenic plants.