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The Effect of EDTA and Fetal Cord Serum Supplementation to Ham's F-10 Culture Medium on Developmental Potential of Mouse Embryos In Vitro (Ham's F-10 배양액에 첨가된 태아제대혈청과 EDTA가 백서 수정란의 분할에 미치는 영향)

  • Kim, Byeong-Seog;Lee, Young-Gi;Park, Yoon-Kee;Lee, Tae-Hyung;Lee, Sung-Ho
    • Journal of Yeungnam Medical Science
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    • v.12 no.1
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    • pp.124-134
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    • 1995
  • It is the most important to select optimal culture conditions to promote safe embryo growth in the technique of human in vitro fertilization and embryo transfer. It has been shown that the addition of biologic fluids, such as blood serum, of various origins, improved fertilization and early cleavage rates in numerous species. The purpose of this study is to attempt to measure developmental potential of mouse eggs fertilized and cleaved in Ham's F10 culture medium containing a chelating agent, EDTA and fetal cord serum. In this study, we selected 40 female mice and 20 male mice, and investigated optimal serum concentrations for mouse embryo growth. Two cell stage mouse embryos were cultured in Ham's F-10 medium, Ham's F-10 medium with various concentrations of EDTA, or Ham's F-10 medium with EDTA and 10% human cord serum. Developmental ratios to morula in Ham's F-10 medium containing various concentrations of EDTA and/or 10% fetal cord serum were significantly higher than in unsupplemented Ham's F-10 medium (p<0.05). Developmental ratios to blastocyst in Ham's F-10 containing 10% fetal cord serum and $50{\mu}M$ or $100{\mu}M$ EDTA were significanltly higher than in unsupplemented Ham's F-10 medium (p<0.05). Developmental ratios to morula in Ham's F-10 containing 10% fetal cord serum and $100{\mu}M$ EDTA were significanltly higher than in Ham's F-10 with 10% fetal cord serum used commonly in many human IVF centers(p<0.05). Developmental ratio to blastocyst in Ham's F-10 containing 10% fetal cord serum and $100{\mu}M$ EDTA was significanlty higher than in Ham's F-10 with $200{\mu}M$ EDTA(P<0.05). In summary, embryo development to morula and blastocyst was significanlty higher in the presence of human cord serum or EDTA than in the unsupplemented medium. The most significanly development to morula and blastocyst was obtained at Ham's F-10 medium with $100{\mu}M$ concentration of EDTA and 10% fetal cord serum. These results suggest that Ham's F-10 medium containing 10% fetal cord serum and optimal concentrations of EDTA significantly promoted early cleavage of mouse zygotes, and these will be useful as basic data for the selection of culture medium in human in vitro fertilization.

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Anti-inflammatory Activity of Antimicrobial Peptide Protaetiamycine 2 Derived from the Protaetia brevitarsis seulensis (흰점박이꽃무지 유래 항균 펩타이드 프로테티아마이신 2의 항염증활성)

  • Lee, Joon Ha;Baek, Minhee;Lee, Hwa Jeong;Kim, In-Woo;Kim, Sun Young;Seo, Minchul;Kim, Mi-Ae;Kim, Seong Hyun;Hwang, Jae Sam
    • Journal of Life Science
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    • v.29 no.11
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    • pp.1218-1226
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    • 2019
  • The white-spotted flower chafer Protaetia brevitarsis seulensis is a medicinally beneficial and important edible insect species. We previously performed an in silico analysis of the Protaetia brevitarsis seulensis transcriptome to identify putative antimicrobial peptides and then tested their antimicrobial and hemolytic activities. These peptides had potent antimicrobial activities against bacteria and yeast without inducing hemolysis. In the present study, the cationic antimicrobial peptide, protaetiamycine 2, was selected for further assessment of its anti-inflammatory properties in mouse macrophage Raw264.7 cells. Protaetiamycine 2 treatment of Raw264.7 cells suppressed LPS-induced nitric oxide production and reduced the expression of inducible nitric oxide synthase and cyclooxygenase-2, as determined by real-time PCR and western blotting. The expression of proinflammatory cytokines ($TNF-{\alpha}$, IL-6, and $IL-1{\beta}$) was also attenuated through the MAPKs and $NF-{\kappa}B$ signaling. We also confirmed that protaetiamycine 2 bound to bacterial cell membranes by a specific interaction with LPS. Collectively, these data obtained from LPS-induced Raw264.7 cells indicated that protaetiamycine 2 could have both antimicrobial and anti-inflammatory properties.

The Effect of Kaempferol, guercetin on Hyaluronan-Synthesis Stimulation in Human Keratinocytes (HaCaT) (인체 피부 세포주 (HaCaT)에서 Kaempferol, Quercetin의 Hyaluronan 합성 촉진 효과에 대한 연구)

  • Kim, Seung-Hun;Nam, Gae-Won;Kang, Byung-Young;Lee, Hae-Kwang;Moon, Seong-Joon;Chang, Ih-Seop
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.31 no.1 s.49
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    • pp.97-102
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    • 2005
  • One of the key molecules involved in skin moisture is hyaluronan (hyaluronic acid, HA) with its associated water of hydration. The predominant component of the ECM (extracellular matrix) of skin is HA. It Is the primordial and the simplest of the GAGs (glycosaminoglycans), a water-sorbed macromolecule In extracellular matrix, Included between the vital cells of epidermis. In the skin, HA was previously thought to derive extlusively from dermis. But, recent studies revealed that HA could be synthesized in epidermis. Flavonoids are polyphenolic compounds that is found mainly in foods of plant origin. Kaempferol was known to increase glutathione synthesis in human keratinocyte. And quercetin blocked PPAR-meidated keratinocyte differentiation as lipoxygenase inhibitors. In this study, we sought to evaluate the effect of flavonid, kaempferol and quercetin on production HA in keratinocyte. We examined the changes of three human hyaluronan synthase genes (HASI, HAS2, HAS3) expression by semi-quantitative RT-PCR when kaempferol or quercetin was added to cultured human keratinocytes. We found that these flavonoids slightly upregulated HAS2, HAS3 mRNA after 24 h. And we investigated the effect on HA production by ELISA. When we evaluated the level of HA in culture medium after 24 h incubation. We found enhanced accumulation of HA in the culture medium. Although the effects of above flavonoids are less than retinoic acid, the data indicate that kaempferol, quercetin can dose-dependently increase the level of HA in epidermis cell line. It suggested that flavonoid, kaempferol, and quercetin increased production of HA in skin and it helped to elevate skin moisture and improve facial wrinkle.

Effect of the extracts from Schisandra chinensis Fruit and Morus alba Leaf on Insulin Secretion in Glucose-induced HIT-T15 Cells (오미자와 뽕잎 추출물이 glucose에 의해 유도된 HIT-T15세포의 인슐린 분비능에 미치는 영향)

  • Jeong, Yoo-Seok;Hong, Joo-Heon;Jung, Hee-Kyoung
    • Food Science and Preservation
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    • v.18 no.6
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    • pp.1002-1008
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    • 2011
  • This study aimed to examine the effect of the Schizandra chinensis fruit and Morus alba leaf on insulin expression in HIT-T15 cells, which is exposed by glucose. The total polyphenol contents of the S. chinensis fruit ethanol extract and the M. alba leaf hot-water extract were $20.11{\pm}0.35$ mg/g and $50.02{\pm}0.62$ mg/mL, respectively. The S. chinensis fruit ethanol extract and the M. alba leaf hot-water extract contained $2.85{\pm}0.15$ and $8.76{\pm}0.43$ mg/g flavonoids, respectively. The antioxidant ability of the M. alba leaf hot-water extract was found to be superior to that of the S. chinensis fruit ethanol extract. Compared to the HIT-T15-treated 10 mM 2-deoxy-D-glucose, the $100{\mu}g/mL$ S. chinensis ethanol extract was found to have a two fold increase in insulin productivity. Moreover, the $100{\mu}g/mL$ M. alba leaf hot-water extract promoted the insulin secretion of high-glucose-damaged HIT-T15 almost ten fold. The above results showed that the S. chinensis fruit ethanol extract and M. alba leaf hot-water extract may improve the insulin productivity of the beta cell with glucose-induced oxidative damage. These data suggest that the S. chinensis fruit ethanol extract and the M. alba leaf hot-water extract can be used as food materials for the regulation of insulin secretion.

Breeding of new variety Pleurotus pulmonarius using protoplast fusion technique (원형질체융합 기법을 이용한 산느타리 계통육성)

  • Gwon, Hee-Min;Lee, Yun-Hae;Kim, Jeong-Han;Baek, Il-Sun;Kang, Hee-Wan;Choi, Jong-In
    • Journal of Mushroom
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    • v.19 no.3
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    • pp.166-175
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    • 2021
  • In this study, monokaryons of "Heukari" (Pleurotus ostreatus) and "Hosan" (Pleurotus pulmonarius) were separated to remove the cell wall, and a cross-species protoplast fusion was developed through chemical treatment with polyethylene glycol. The protoplast-fused PF160306 and PF160313 strains have a culture period of 10 and 2 days shorter than that of the "Heuktari" and "Hosan" cultivars, respectively. Furthermore, the growth of the strains was faster than that of the existing cultivars. The yield was 135.9 g per bottle, which was approximately 8% higher than that of the commercially available "Hosan" cultivar; however, it was not statistically significant. A growth survey was conducted after treatment at five temperatures (15, 18, 21, 23, and 25℃). The growth of the strains accelerated with the increase in temperature. However, at 21℃, the yellow color of pileus was the brightest. Band pattern, assessed using URP Primer 7, was similar to that of the "Hosan" cultivar. The DPPH radical scavenging capacity and polyphenol content were 62.5% and 43.5 mg/mL, respectively, for "Sunjung" and 65.7% and 49.9 mg/mL, respectively, for PF160313. Furthermore, the antihypertensive activities of the "Sunjung" cultivar and PF160313 were similarly high at 74% and 75%, respectively. In conclusion, cross-species hybridization via the protoplast fusion technique can be used for obtaining primary data for mushroom breeding to develop new varieties. In addition, the protoplast fusion technique might aid in expanding the market for yellow mushrooms.

Comparison of Microscopy and Pigment Analysis for Determination of Phytoplankton Community Composition: Application of CHEMTAX Program (식물플랑크톤 군집조성 파악을 위한 현미경관찰법과 지표색소분석법 비교 연구: CHEMTAX 프로그램 활용)

  • Kim, Dokyun;Choi, Jisoo;Oh, Hye-Ji;Chang, Kwang-Hyeon;Choi, Kwangsoon;Shin, Kyung-Hoon
    • Korean Journal of Ecology and Environment
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    • v.54 no.4
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    • pp.303-314
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    • 2021
  • To understand how to efficiently observe the biomass and community of phytoplankton, phytoplankton sampling was carried out from June to October 2019 at the Yeongju dam sediment control reservoir(YJ) and Bohyeonsan dam reservoir(BH1 and BH2). The results derived from microscopic observation, such as the conventional phytoplankton qualitative/quantitative analysis, and from the CHEMTAX method based on the pigments, were compared. The relative contribution of phytoplankton, calculated by the microscopy and CHEMTAX methods, showed a significant difference in all four classes: cryptophyta, chlorophyta, cyanobacteria, and diatoms. In addition, the correlation between the two observation methods was poor. This might be caused by methodological differences in microscopy that do not consider the varying cell sizes among phytoplankton species. In this study, by converting the cells into carbon, the slope between both carbon biomasses based on microscopy and CHEMTAX was improved close to the 1 : 1 line, and the y-intercept was closer to 0 for cryptophyta and diatoms. For cyanobacteria, the slope increased, the y-intercept decreased, and the plot approached 1 : 1 although the correlation coefficients were not improved in all classes. The present study suggests that application of CHEMTAX based on pigment analysis could be a possible approach to efficiently determine the relative carbon proportions of individual classes of phytoplankton community composition.

Grapevine Growth and Berry Development under the Agrivoltaic Solar Panels in the Vineyards (영농형 태양광 시설 설치에 따른 포도나무 생육 및 과실 특성 변화 비교)

  • Ahn, Soon Young;Lee, Dan Bi;Lee, Hae In;Myint, Zar Le;Min, Sang Yoon;Kim, Bo Myung;Oh, Wook;Jung, Jae Hak;Yun, Hae Keun
    • Journal of Bio-Environment Control
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    • v.31 no.4
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    • pp.356-365
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    • 2022
  • Agrivoltaic systems, also called solar sharing, stated from an idea that utilizes sunlight above the light saturation point of crops for power generation using solar panels. The agrivoltaic systems are expected to reduce the incident solar radiation, the consequent surface cooling effect, and evapotranspiration, and bring additional income to farms through solar power generation by combining crops with solar photovoltaics. In this study, to evaluate if agrivoltaic systems are suitable for viticulture, we investigated the microclimatic change, the growth of vines and the characteristics of grape grown under solar panels set by planting lines compared with ones in open vineyards. There was high reduction of wind speed during over-wintering season, and low soil temperature under solar panel compared to those in the open field. There was not significant difference in total carbohydrates and bud burst in bearing mother branches between plots. Despite high content of chlorophyll in vines grown under panels, there is no significant difference in shoot growth of vines, berry weight, cluster weight, total soluble solid content and acidity of berries, and anthocyanin content of berry skins in harvested grapes in vineyards under panels and open vineyards. It was observed that harvesting season was delayed by 7-10 days due to late skin coloration in grapes grown in vineyards under panels compared to ones grown in open vineyards. The results from this study would be used as data required in development of viticulture system under panel in the future and further study for evaluating the influence of agrivoltaic system on production of crops including grapes.

Synthesis and Structural Study of Extraframework ZrI6Tl119+ Cationic Cluster in Zeolite A (제올라이트 A 동공 내 비골격 ZrI6Tl119+ 양이온 클러스터의 합성과 구조 연구)

  • Hyeon Seung, Lim;Jong Sam, Park;Cheol Woong, Kim;Woo Taik, Lim
    • Korean Journal of Mineralogy and Petrology
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    • v.35 no.4
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    • pp.447-455
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    • 2022
  • Fully dehydrated Tl12-LTA (|Tl12|[Si12Al12O48]-LTA,Tl12-A) was treated with 6.0×103 Pa of ZrI4 (g) at 623 K for 72 hr under anhydrous conditions. The crystal structure of product, |Zr0.25I1.5Tl12|[Si12Al12O48]-LTA, was determined by single-crystal crystallography using synchrotron X-radiation in the cubic space group Pm3m (a = 12.337(2) Å). It was refined using all data to the final error index (for the 712 unique reflections for which Fo> 4σ(Fo) R1/wR2= 0.055/0.189. In this structure, octahedral ZrI62- ions center about 25% of the large cavities (Zr-I = 2.91(4) Å). Each coordinates to eight Tl+ ions and they are further bridged by Tl+ ions in the planes of 8-rings to form a cubic three-dimensional ZrI6Tl119+ cationic cluster. About 1.5 Tl+ ions per unit cell moved to deeper side of sodalite cavity after reaction with ZrI4(g). The remaining Tl+ ions occupy well-established cation positions near 6- and 8-rings.

Physiological Responses of Common Carp (Cyprinus Capio) and Crucian Carp (Carassius Auratus) by Rapid Changes of Water Temperature (급격한 수온변화에 따른 잉어 (Cyprinus capio)와 붕어 (Carassius auratus)의 생리적 반응)

  • Moon, Jeong Suk;Hur, Jun Wook
    • Ecology and Resilient Infrastructure
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    • v.9 no.2
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    • pp.113-123
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    • 2022
  • The blood and physiological changes of common carp (Cyprinus carpio) and crucian carp (Carassius auratus) were analyzed when the water temperature was rapidly increased from 20℃ (control) to 26 and 32℃. The water temperature reached 26℃ and 32℃ within 6 hours from the water temperature of 20℃, and the control was maintained at 20℃ for the duration of the experiment. From each experimental group, blood was collected every 3, 12, 24 and 48 hours after the water temperature rise, and the red blood cell count, hemoglobin, hematocrit, Na+, K+, Cl-, cortisol, glucose, aspartate amino transferase (AST) and alanine amino transferase (ALT) contents were analyzed. In the case of the crucian carp experimental group where the water temperature was raised to 32℃, the concentrations of plasma cortisol, glucose, AST and ALT increased 3 hours after the water temperature rise, and the concentrations did not decrease until 48 hours (P<0.05). Plasma cortisol, glucose, AST, and ALT in both C. carpio and C. auratus in the control group and the experimental group raised to 26℃ increased at 3 hours after the water temperature rise (P<0.05), and decreased to the value before the water temperature rise at 48 hours (P<0.05). The plasma Na+, K+ and Cl- concentrations of C. carpio and C. auratus in the experimental group where the water temperature was raised to 32℃ were higher than in the 20 and 26℃ experimental groups. The blood reaction of all experimental groups including the control group showed a common trend in both C. carpio and C. auratus increasing up to 12 hours after the water temperature rise and then decreasing at 48 hours after the water temperature rise. The results derived from this study would be useful for investigating the physiological response of fish stress in future. In addition, as fish mortality has recently occurred frequently in rivers and lakes, it is judged that it can be used as basic data.

Cryopreservation of Bovine IVM/IVF/IVC Hatched Blastocysts (체외생산된 소 완전탈출 배반포기배의 동결보존)

  • Lee, K.S.;Kim, E.Y.;Yi, B.K.;Nam, H.K.;Yoon, S.H.;Park, S.P.;Lim, J.H.
    • Korean Journal of Animal Reproduction
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    • v.23 no.2
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    • pp.141-148
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    • 1999
  • This study was to test whether the viability of bovine hatched blastocysts (HBs) can be maintained after vitrification and thawing. The HBs were produced in vitro at Day 9 and Day 10 after IVF, and they were classified to small (S-HBs; ø$\leq$300 ${\mu}{\textrm}{m}$) and large(L-HBs; ø>300 ${\mu}{\textrm}{m}$) on the basis of embryo diameter using eyepiece micrometer. As freezing solution, we used EFS35 which consisted of 35% ethylene glycol (EG), 18% ficoll, 0.3 M sucrose and 10% FBS added in mDPBS. Vitrification was taken by two-step method, the HBs were equilibrated in 10% EG for 5 minutes and then shortly exposed in EFS35 and plunged into L$N_2$for 30~45 sec. After thawing, the survival rates were assessed by the re-expansion of the blastocoel during 2 h and 16 h of culture. The results obtained in these experiments were summarized as follows; 1) When the blastocysts(40.8%) recovered at Day 8 after IVF were further cultured for 24 h(Day 9 after IVF) and 48 h(Day 10 after IVF), the rates of HBs were 20.5% and 6.7%, respectively. Also, the total cell number of HBs on Day 9 was significantly higher than that of HBs on Day 10 (p<0.01). 2) When the effects of freezing solution to the survival of Day 9 L-HBs were examined, the rate of vitrified group (75.7%) was significantly lower than 100% of control and exposed group(p<0.05). 3) When the survival rates of vitrified HBs according to size and developmental age were examined, the data of L-HBs (75.5%) and S-HBs(63.6%) on Day 9 were slightly higher than those of L-HBs(64.3%) and S-HBs(60.7%) on Day 10. 4) Also, when the in vitro survival of Day 9 HBs was evaluated under different culture condition after thawing, the result in culture medium only (79.3%) was significantly higher than 43.2% in co-culture group (p<0.05). These results demonstrated that bovine HBs can be successfully cryopreserved by two-step vitrification method using EFS35.

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