• Title/Summary/Keyword: Liquid membrane

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Current and Future Perspectives of Lung Organoid and Lung-on-chip in Biomedical and Pharmaceutical Applications

  • Junhyoung Lee;Jimin Park;Sanghun Kim;Esther Han;Sungho Maeng;Jiyou Han
    • Journal of Life Science
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    • v.34 no.5
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    • pp.339-355
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    • 2024
  • The pulmonary system is a highly complex system that can only be understood by integrating its functional and structural aspects. Hence, in vivo animal models are generally used for pathological studies of pulmonary diseases and the evaluation of inhalation toxicity. However, to reduce the number of animals used in experimentation and with the consideration of animal welfare, alternative methods have been extensively developed. Notably, the Organization for Economic Co-operation and Development (OECD) and the United States Environmental Protection Agency (USEPA) have agreed to prohibit animal testing after 2030. Therefore, the latest advances in biotechnology are revolutionizing the approach to developing in vitro inhalation models. For example, lung organ-on-a-chip (OoC) and organoid models have been intensively studied alongside advancements in three-dimensional (3D) bioprinting and microfluidic systems. These modeling systems can more precisely imitate the complex biological environment compared to traditional in vivo animal experiments. This review paper addresses multiple aspects of the recent in vitro modeling systems of lung OoC and organoids. It includes discussions on the use of endothelial cells, epithelial cells, and fibroblasts composed of lung alveoli generated from pluripotent stem cells or cancer cells. Moreover, it covers lung air-liquid interface (ALI) systems, transwell membrane materials, and in silico models using artificial intelligence (AI) for the establishment and evaluation of in vitro pulmonary systems.

Effect of Different Infusion Frequency of Liquid Nitrogen on Actin Filament, Mitochondria, Apoptosis and Development in Mouse 2-Cell Embryo after Freezing and Thawing (생쥐 배아 동결시 액체질소의 분사속도가 해빙후 배아의 발달, 미세섬유, 미토콘드리아 및 세포자연사에 미치는 영향)

  • 손인표;안학준;계명찬;최규완;민철기;강희규;이호준;권혁찬
    • Development and Reproduction
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    • v.4 no.2
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    • pp.161-173
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    • 2000
  • The aim of this study was to assess the effect of the frequency of the L$N_2$ infusion on the ultrastructure, metabolism and development of the embryo after freezing and thawing by computerized cell freezer. Two-cell embryos of ICR mouse were randomly allocated into fresh (control), high-frequency freezing (group 1) and low-frequency freezing (group 2). For fresh and frozen-thawed intact 2-cell embryos, total ceil number in the blastocyst was counted by fluorescent microscope after Hoechst 33258 staining. Relative amount of $H_2O$$_2$ was measured by DCHFDA. Intracellular location and membrane potential of the mitochondria were evaluated by staining with rhodamine 123 and JC-1. The structure of actin filament was also evaluated by confocal microscope. DNA fragmentation was assessed by TUNEL method after development into blastocyst. The survival rate of intact embryo was higher in group 1 than group 2 (50.7% vs. 34.6% respectively, p<0.05). The blastocyst developmental rate was significantly low in group 2 (86.7%, 76.7% vs. 44.0% for control, group 1 and group 2 respectively, p<0.05). Total cell number in the blastocyst was also significantly lower in group 2 than control (79.5$\pm$12.9, 71.6$\pm$8.0, and 62.5$\pm$4.7 for control, group 1 and group 2 respectively, p<0.05). The relative amount of $H_2O$$_2$ was higher in group 2 than other groups (15.3$\pm$3.0, 16.6$\pm$1.6 vs. 23.4$\pm$1.8, p<0.05). After JC-1 staining, relative intensity of mitochondria with high membrane potential was significantly lower in group 2 than control and group 1 (17.2$\pm$3.8, 17.4$\pm$1.3 vs. 13.2$\pm$2.0, p<0.05). In group 2, partial deletion and aggregation of the actin filament was found. DNA fragmentation rate was also hieher for group 2 versus other groups (30.8%, 36.0% vs. 65.6%, p<0.05). The frequency of the L$N_2$ infusion is an important factor for the development of frozen-thawed mouse embryo. High-frequency infusion may prevent damages of cytoskeleton and mitochondria in the embryo probably by preventing the temperature fluctuation during dehydration phase. We speculate that the application of high-frequency infusion method in human embryo may be promising.

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Effects of Kinds of Cryoprotectants on the Characteristics of Frozen Fowl Semen (닭 정액 동결 시 동결 보호제가 정액 성상에 미치는 영향)

  • Choi, Jin Seok;Shin, Dan-Bi;Ko, Yeoung-Gyu;Do, Yoon-Jung;Byun, Mijeong;Park, Soo-Bong;Seong, Hwan-Hoo;Kim, Hyun;Kong, Il-Keun;Kim, Sung Woo
    • Korean Journal of Poultry Science
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    • v.40 no.3
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    • pp.171-178
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    • 2013
  • The purpose of this study was to evaluate the sperm viability, normal acrosome and mitochondrial activity in the frozen-thawed fowl semen by different cryoprotectants. The experiment was carried out on 10 sexually adult roosters of Ogye. The semen was collected twice a week and pooled semen was diluted 1:1 EK extender containing no cryoprotectant at $5^{\circ}C$. After equilibration for 30 minutes, diluted chicken semen was diluted 1:1 extender containing either 7% dimethylacetamide (DMA), 7% dimethylformamide (DMF) or 7.5% methylacetamide (MA) at final concentration and was put in 0.5 mL plastic straws and frozen for 30 minutes by exposure to liquid nitrogen vapor 4 cm above the surface of liquid nitrogen, followed by plunging into liquid nitrogen. Frozen semen was thawed in water bath at $5^{\circ}C$ for 2 minutes. For cytometric analysis, the frozen-thawed semen was diluted with EK extender to a final concentration of 90 million spermatozoa per mL. Sperm membrane integrity was evaluated as SYBR-14 and propidium iodide (PI). Acrosome integrity was assessed with fluorescein isothiocyanate-labeled PSA and PI. The percentage of mitochondrial function was estimated by using Rhodamine123 (R123) and PI. In conclusion, freezing rooster semen by using 7% DMF as cryoprotectant was significantly highest in rates of survival and mitochondrial function while its rate of damage of acrosome was significantly lowest. As a result, DMF is the cryoprotectant that has the lowest influences on sperm membranes and acrosome integrity. Therefore it could be used for freezing method of animal genetic conservation method for poultry diversity.

Emulsion Liquid Membrane Transport of Heavy Metal Sons by Macrocyclic Carriers (거대고리 운반체에 의한 중금속이온의 에멀죤 액체막 수송)

  • 정오진
    • Journal of Environmental Science International
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    • v.4 no.2
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    • pp.223-232
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    • 1995
  • New two macrocyclic compounds using as carriers of liquid emulsion menbrame, have been synthesized. These reuslts provide evidance for the usefulness of the theory in designing the systems. The efficiency of selective transport for heavy metal ions have been discussed from the membrane systems that make use of $SCN^-$,<>,$I^-$,CN- and $Cl^-$ ion as co-anions in source phase and make use of $S_2O_3^{2-}$ and $P_2O_7^{4-}$ ion as receiving phase, respectively. The transport rate of M(II) was highest when a maximum amount of the M(II) in the source phase was present as$Cd(SCN)_2$$(P[SCN^-]= 0.40M)$, $Hg(SCN)_2([SCN^-]=0.40M)$ and Pd(CN)$([CN^-]= 0.40M)$. The Cd(II) and Pb(II) over each competitive cations were well transprted with 0.3M-S2032- and 0.3M-P2O74-, respectively in the receiving phase. Results of this study indicate that two criteria must be met in order to have effective macrocycle-mediated transport in these emulsion system. First one must effective extraction of the $M^{n+}$ into the toluene systems. The effectiveness of this extraction is the greatest if locK for $M^{n+}$macrocycle interaction is large and if the macrocycle is very insoluble in the aqueous phase. Second, the ratio of the locK values (or Mn+-receiving phase ($S_2O_3^{2-}$- or $P_2O_7^{4-}$) to $M^{n+}$-macrocycle (($L_1$이나 $L_2$) interaction must be large enough to ensure quantitative stripping of Mn+(($Cd^{2+}$,$Pb^{2+}$)at the toluene receiving Phase interface. $L_1$(3.5-benzo-10,13,18,21-tetraoxa-1,7,diazabicyclo(8,5,5) eicosan) forms a stable ($Cd^{2+}$ and >,$Pb^{2+}$ complexes and $L_1$ is very insoluble in water and its $Cd^{2+}$ and >,$Pb^{2+}$ complex is considerably less stable than $Cd^{2+}$-(S2O3)22- and $Pd^{2+}-P_2O_7^{4-}$ complexes. On the other hand, the stability of the $Hg^{2+}$)+-$L_1$( complex exceed that of the $Hg^{2+}$- (S2O3)22- and Hg2+-P2O74-, and the distribution coefficient of $L_2$(5,8,15,18,23,26-hexaoxa-1,12- diazabicyclo-(10,8,8) octacosane) is much smaller than that of $L_1$. Therefore, the partitioning of Lr is favored by the aqueous receiving Phase, and little heavy metal ions transport is seen despite the large logK for $Hg^{2+}$+-$L_1$ and $Mn^+$($Cd^{2+}$+, $Pb^{2+}$+ and $Hg^{2+}$)-$L_2$ interactions. Key Words : macrocycles, transport, heavy metal, co-anion, source phase, receiveing, complex separation, interaction, destribution coefficient.

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Adsorption Behavior of Pb2+ Ions on Alginate Beads and Capsules (알지네이트 비드와 캡슐에서의 납 이온의 흡착거동)

  • Shin, Eun Woo;Thuong, Nguyen Thi Lien;Yoo, Ik-Keun
    • Korean Chemical Engineering Research
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    • v.45 no.2
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    • pp.166-171
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    • 2007
  • The adsorption behavior of $Pb^{2+}$ was compared between calcium alginate beads and capsules, which have different structures of alginate-gel core beads and liquid core alginate-membrane capsules, respectively. In terms of adsorption kinetics and isotherms, adsorption characteristics depending on pH and hardening time were compared for both adsorbents and also released calcium ion during the adsorption process was monitored. The adsorption of $Pb^{2+}$ on both adsorbents was caused by surface complexation and ion exchange mechanisms, both of which have similar effects on adsorption process regardless of the amount of adsorbed $Pb^{2+}$. The dependence of $Pb^{2+}$ adsorption upon pH was also similar for both adsorbents indicating the existence of similar functional groups on the surface of adsorbents. However, a different $Pb^{2+}$ adsorption behavior was observed considering the adsorption kinetics. The adsorption kinetic of $Pb^{2+}$ on alginate beads was slower than on alginate capsules and the maximum adsorption loading ($Q_{max}$) onto alginate beads was also less than onto alginate capsules by 49%. This drawback of alginate beads compared to capsules were ascribed to a diffusion limitation due to solid gel-core structure of alginate beads.

Association study analysis of phospholipase C zeta gene polymorphism forsperm motility and kinematic characteristics in liquid semen of Boar (Phospholipase C zeta 유전자의 유전적다형성과 돼지 액상정액의 운동학적 특성과의 연관성 분석)

  • Jeong, Yong-dae;Jeong, Jin-Young;Sa, Soo-Jin;Kim, Ki-Hyun;Cho, Eun-Seok;Yu, Dong-Jo;Park, Sungk-won;Jang, Hyun-Jun;Woo, Jae-Seok;Choi, Jung-Woo
    • Journal of Embryo Transfer
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    • v.31 no.3
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    • pp.293-297
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    • 2016
  • For evaluating the boar semen quality, sperm motility is an important parameter because the movement of sperm indicates active metabolism, membrane integrity and fertilizing capacity. Phospholipase C zeta (PLCz) is important enzyme in spermatogenesis, but the effect has not been confirmed in pigs yet. Therefore, this study was aimed to analyze their association with sperm motility and kinematic characteristics. DNA samples from 124 Duroc pigs with records of sperm motility and kinematic characteristics [total motile spermatozoa (MOT), curvilinear velocity (VCL), straight-line velocity (VSL), the ratio between VSL and VCL (LIN), amplitude of lateral head displacement (ALH)] were subjected. A SNP in non-coding region of PLCz g.158 A > C was associated with MOT (p < 0.05), VCL (p < 0.01), LIN (p < 0.01) and ALH (p < 0.05) in Duroc population. Therefore, we suggest that the intron region of the porcine PLCz gene may be used as a molecular marker for Duroc boar semen quality, although its functional effect was not defined yet. Whether the association is due to the candidate gene or not require further verification. Thus, it will be of interest to continue association studies in the regions surrounding those genes.

Positive Expression of EGFP Gene in Bovine Embryos after ICSI using Spermatozoa Co-cultured with Exogenous DNA (외래 유전자와 공배양한 정자를 이용해 난자내 직접 주입술한 후 EGFP의 발현)

  • 윤효진;이훈택;정길생
    • Korean Journal of Animal Reproduction
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    • v.26 no.3
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    • pp.205-214
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    • 2002
  • There are many methods to introduce exogenous DNA into embryo to produce transgenic animals. Exogenous gene can be integrated into oocyte by sperm vector. In this study, sperm was used as a vector for a transgene, which is encoding enhanced green fluorescent protein (EGFP). The objective of this study was to investigate the expression of exogenous gene in bovine embryos after injection of spermatozoa cocultured with EGFP DNA fragment. Spermatozoa were plunged into liquid nitrogen and thawed several times or shook in 0.2% Triton X-100 to remove sperm membrane followed by DTT treatment. The injected oocytes were co-cultured with vero cells in CR1aa, and expression of EGFP gene was observed under fluorescent microscope. Blastocyst formation rates of oocytes injected with sperm treated with DTT, DTT-freezing or DTT-Triton X-100 were 34.7, 39.4 and 31.9%, respectively. The rates of EGFP expression in oocytes injected with 54 ng DNA after DTT-treated, DTT-freezing and DTT-Triton X-100-treated sperm were 0, 19.1 and 13.9%. On the other hands, expression rate of oocytes injected with sperm cocultured with 13.5, 27 and 63.5 ng of EFGP DNA were 6.7, 9.0 and 5.1%, respectively. When intact sperm was mixed with 63.5 ng/${mu}ell$ EGFP DNA fragment, and then electroporated before injection, the expression rate of injected oocyte was 2%. Unexpectedly, electro-poration could not increase the expression rate. These results suggest that sperm can be used as a transgene vector, even if the efficiency was low (19.1%).

Distribution of Cadmium in a Strain of Staphylococcus aureus Resistant Against the Metal (카드뮴내성(耐性) Staphylococcus aureus내(內) 카드뮴분포(分布))

  • Hyun, Eun-Min;Park, Chan-Seung;Choi, Kyoung-Ho
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.10 no.1
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    • pp.103-106
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    • 1981
  • A strain of Staphylococcus aureus resistant against cadmium was cultivated by using a liquid medium containing 10ppm cadmium ion, and then, it was fractionated into several fractions as described in the previous paper. Content of the metal in each fraction was determined through an atomic absorption spectrometery. The results are as follows; (1) A $690.9{\mu}g$ cadmium was contained in one gram dry cell. (2) A 39.9% of total cadmium was easily extracted by TCA, however a 52.2% was unextractable even by series of extraction with TCA, ethanol-ehter, perchloric acid and ammonium water. (3) Among the fractions prepared along the cellular structure, plasma membrane fraction showed a highest content of the metal by containing a 59.1%. (4) The fraction of cytoplasm and cell wall contained a 26.8 and 14.1%, respectively. (5) More than 90% of the metal contained in the cell wall was detected from the fraction of lipopolysaccharide. It is considered from these results tht at least a 70% of the cadmium up taken by the resistant cell associates with membranous structure in the cell surface.

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Effects of Chronic Ethanol Consumption and Taurine Supplementation on Hepatic Total and Phospholipid Fatty Acid Compositions in Rats (만성적인 에탄올 섭취와 타우린보강이 흰쥐간의 총지방산 및 인지질지방산 조성에 미치는 영향)

  • Um, Young-Sook;Chung, Eung-Jung;Oh, Joo-Yeon;Park, Tae-Sun
    • Journal of Nutrition and Health
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    • v.33 no.2
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    • pp.124-133
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    • 2000
  • This study evaluated the effects of chronic ethanol consumption and/or taurine supplementation on hepatic total, phospholipid fatty acid composition and the metabolism of rats fed one of three purified liquid diets for 8 weeks. the rats followed either the control diet (CD, ethanol-free and taurine-free diet); ethanol diet (ED, CD+ 50g ethanol/L) or ethanol-taurine diet (ETD, ED+3.75g taurne/L). Chronic ethanol consumption and/or dietary taurine supplementation were associated with altered hepatic total and phospholipid fatty acid composition. compared to the values for the control rats, ED or ETD significantly decreased the percentage of total monounsaturated fatty acids ($\Sigma$MUFA), and increased the percentage of total polyunsaturated fatty acids ($\Sigma$PUFA) of hepatic total lipids(p〈0.01). Percentages of 14:0(P〈0.01) and 16:0(p〈0.001) were sigificantly lower, and those of 18:0(p〈0.01), 20:0(p〈0.001), 20:3$\omega$6(p〈0.01) and 22:4$\omega$6(p〈0.01) in hepatic total fatty acid compositions were oserved in rats fed ETD versus those fed ED or ETD. No significant differences in hepatic total fatty acid compositions were observed in rats fed ETD versus those fed ED. Percentages of 24:0(p〈0.01), 16:1(p〈0.05), 20:1(p〈0.01), 18:2$\omega$6(p〈0.01) and 18:3$\omega$3(p〈0.05) in hepati phospholipids were significantly higher, and those of 14:0(p〈0.01), 16:0(p〈0.001), 20:3$\omega$3(p〈0.05) in hepatic phospholipids were significantly higher, and those of 14:0(p〈0.01), 16:0(p〈0.001), 20:3$\omega$3(p〈0.001), 22:6$\omega$3(p〈0.001) and $\Sigma$$\omega$3(P〈0.001) were significantly lower in rats fed ED or ETD compared to the values for the control rats. The Δ5 desaturation index(20:3$\omega$6⇒20:4$\omega$6) and elongation index (20:5$\omega$3⇒22:5$\omega$3) of hepatic phospholipid index (20:3$\omega$6⇒20:4$\omega$6) and decreased Δ4 desaturation index (22:5$\omega$3⇒22:6$\omega$3) compared to the values for the ED rats. These changes in hepatic fatty acid composition induced by chronic ethanol consumption and/or taurine supplementation might be associated with the modulations of physical properties of the hepatic cell membrane and its sensitivity to peroxidation damage.

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A Study on Paralytic Shellfish Poison of Sea Mussel, Mytilus edulis -Food Poisoning Accident in Gamchun Bay, Pusan, Korea, 1986- (진주담치의 마비성독에 관한 연구 -1986년 부산 감천만 중독사고를 중심으로-)

  • CHANG Dong-Suck;SHIN Il-Shik;PYEUN Jae-Hyeung;PARK Yeung-Ho
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.20 no.4
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    • pp.293-299
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    • 1987
  • At various times and places all over the world men have become ill and some have died after eating shellfish that were intoxicated with paralytic shellfish poison(PSP) caused by Protogonyaulax spp. In late March, 1986, two persons were dead by ingesting wild sea mussels, Mytilur edulis, grown at bottom of an anchored waste ship to be dismantled at Gamchun Bay, Pusan, Korea. The samples were collected from the bottom of the ship during April $1\~April$ 8 of the year to find the cause of the food poisoning accident. The toxicity was estimated by bioassay with ICR male mouse, while the toxins were extracted and characterized. The toxins were extracted with acidified $80\%$ ethanol. The extract was defatted three times with dichloromethane, treated with activated charcoal, and then purified by chromatography on Bio-Gel P-2 and Bio-Rex 70. The toxic fractions obtained were analysed by cellulose acetate membrane electrophoresis, thin layer chromatography and high performance liquid chromatogaphy. The range and the average of PSP-toxicity of the samples were $132\~295\;MU/g$, 203 MU/g respectively. The amount of PSP was $26.4\~58.9{\mu}g/g$ of whole meat in range and $40.6{\mu}g/g$ in average. The toxicity of the digestive gland of the samples was 9 times higher than that of edible meat (except digestive gland) as $439\~979MU/g$, and it was about $70\%$ in total toxin. The compositional analytical results of the paralytic shellfish toxin, Gonyautoxin $1\~4$ were the major part of the PSP and Saxitoxin and neosaxitoxin were detected as the minor component. It was concluded that the food poisoning accident was caused not by Saxitoxins but by Gonyautoxins.

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