• Title/Summary/Keyword: liquid nitrogen

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In vitro Follicular Growth and Ovulation of Mouse Preantral Follicles Cryopreserved by Vitrification (초자화동결된 생쥐 Preantral Follicle의 체외성장과 배란)

  • Park, Ji-Kwon;Paik, Won Young
    • Clinical and Experimental Reproductive Medicine
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    • v.32 no.2
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    • pp.91-99
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    • 2005
  • Objective: To define an appropriate vitrification condition of preantral follicle that yields high survival and to evaluate growth and ovulation rate of mouse follicles during in vitro culture after vitrification. Methods: Preantral follicles were isolated mechanically from mouse ovaries that were surgically recovered from mice aged 14 days. Retrieved preantral follicles were placed in EG (Ethylene Glycol) for 2, 5, 10 minutes and transferred to EFS-40 (40% EG, 18% Ficoll-70, 0.5 M sucrose) for 0.5, 1, 2 minutes. And then, preantral follicles were placed onto an EM grid and submerged immediately in liquid nitrogen. Thawing was carried out at room temperature. After defining the most appropriate vitrification condition that yields high survival, in vitro growth and ovulation rate of follicles were evaluated. Results: Appropriate vitrification condition that yield high survival rate ($83.2{\pm}2.1%$) of preantral follicle was EG for 5 minutes and EFS-40 for 0.5 minutes. In vitro survival rate of the vitrified preantral follicles were $85.5{\pm}0.5%$, $67.9{\pm}0.8%$ and $40.2{\pm}0.5%$ on day 2, 6 and 10. And in vitro growth of the vitrified preantral follicles were $107.1{\pm}16.1{\mu}m$, $117.1{\pm}18.4{\mu}m$, $178.4{\pm}45.6{\mu}m$ and $325.4{\pm}54.4{\mu}m$ on day 0, 2, 6 and 10. Although in vitro survival rate and growth of vitrified preantral follicles were lower than that of non-vitrified preantral follicles, the patterns of survival and growth were similar in vitrified and non-vitrified preantral follicles. The ovulation rate of antral follicles that was grown from vitrified preantral follicles was $32.6{\pm}1.2%$. Conclusion: Vitrified preantral follicles could be grown to antral sizes, and mature oocytes that can be used for IVF-ET programs were produced successfully. These data suggest that cryopreservation of preantral follicle by vitrification can be used for the preservation of the fertility.

HPLC Determination of Tolperisone in Human Plasma

  • Bae Jung-Woo;Park Young-Seo;Sohn Uy-Dong;Myung Chang-Sun;Ryu Byung-Kwon;Jang Choon-Gon;Lee Seok-Yong
    • Archives of Pharmacal Research
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    • v.29 no.4
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    • pp.339-342
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    • 2006
  • A simple high performance liquid chromatographic (HPLC) method was developed for the determination of tolperisone in human plasma. Tolperisone and internal standard (chlorphenesin) were isolated from 1 mL of plasma using 8 mL of dichlormethane. The organic phase was collected and evaporated under nitrogen gas. The residue was then reconstituted with 300 mL aliquot of mobile phase and a 100 mL aliquot was injected onto the $C_{18}$ reverse-phased column. The mobile phase, $45\%$ methanol containing $1\%$ glacial acetic acid and $0.05\%$ 1-hexanesulfonic acid was run at a flow rate of 1 mL/min. The column effluent was monitored using UV detector at 260 nm. The retention times for tolperisone and the internal standard were approximately 7.1 and 8.4 min, respectively. The standard curve was linear with minimal intra-day and inter-day variability. The quantification limit of tolperisone in human plasma was 10 ng/ mL. The proposed method has been applied to the determination of pharmacokinetic profile of tolperisone in Koreans. The T max of tolperisone in Koreans $(0.94{\pm}0.42\;h)$ was not significantly differ from that reported in Europeans (0.5-1 h), but the mean half-life in Koreans $(1.14{\pm}0.27\;h)$ was shorter than that in Europeans $(2.56{\pm}0.2\;h)$. The proposed HPLC method is simple, accurate, reproducible and suitable for pharmacokinetic study of tolperisone.

The Properties of Livestock Waste Composts Tea Depending on Manufacturing Method and Their Effect on Chinese Cabbage Cultivation

  • Jang, Jae-Eun;Kang, Chang-Sung;Park, Jung-Soo;Kim, Sun-Jae;Kim, Hee-Dong
    • Korean Journal of Soil Science and Fertilizer
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    • v.48 no.1
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    • pp.8-14
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    • 2015
  • Livestock waste compost tea is a liquid extract of compost obtained by mixing livestock compost. In this study, some chemical and microbiological characteristics of compost tea depending on the kind of raw materials used were examined, and several experiments to investigate the practical effects on Chinese cabbage cultivation were conducted. This experiment showed that livestock composts needed to be added into aerated water at the ratio between 1:100 and 1:10 (1 part compost to 10~100 parts water) to produce the high quality compost tea. Compost teas must be aerated more than 24 to 48 hours to be able to support aerobic organisms. In cultivation test with compost teas, swine manure compost teas were made by the extracting ratio of 50x, in the aerated condition for 24 hours in water and oil cake in the extracting ratio of 100x were added as supplements. Following the input of oil cake, the concentration of nitrogen and aerobic bacteria increased. Another experiment was conducted to determine the effect of different swine manure compost teas on plant growth and yield of Chinese cabbage. The fresh yield of Chinese cabbage was higher in the fertigated plots by compost tea with oil cake compared to those of N, $P_2O_5$, $K_2O$ fertilization plot with chemical fertilizer by soil test recommendation (Fert. NPK). The effect of compost tea on growth of Chinese cabbage was largely attributable to the increased number of microorganisms as well as nutrients.

Saengmaeksan, a traditional herbal formulation consisting of Panax ginseng, ameliorates hyperuricemia by inhibiting xanthine oxidase activity and enhancing urate excretion in rats

  • Sung, Yoon-Young;Yuk, Heung Joo;Kim, Dong-Seon
    • Journal of Ginseng Research
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    • v.45 no.5
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    • pp.565-574
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    • 2021
  • Background: Saengmaeksan (SMS) is a traditional Korean medicine composed of three herbs, Panax ginseng, Schisandra chinensis, and Liriope platyphylla. SMS is used to treat respiratory and cardiovascular disorders. However, whether SMS exerts antihyperuricemic effects is unknown. Methods: Effects of the SMS extract in water (SMS-W) and 30% ethanol (SMS-E) were studied in a rat model of potassium oxonate-induced hyperuricemia. Uric acid concentrations and xanthine oxidase (XO) activities were evaluated in the serum, urine, and hepatic tissue. Using renal histopathology to assess kidney function and uric acid excretion, we investigated serum creatinine and blood urea nitrogen concentrations, as well as protein levels of renal urate transporter 1 (URAT1), glucose transporter 9 (GLUT9), and organic anion transporter 1 (OAT1). The effects of SMS on in vitro XO activity and uric acid uptake were also evaluated. The components of SMS were identified using Ultra Performance Liquid Chromatography (UPLC). Results: SMS-E reduced serum uric acid and creatinine concentrations, and elevated urine uric acid excretion. SMS-E lowered XO activities in both the serum and liver, and downregulated the expression of renal URAT1 and GLUT9 proteins. SMS-E reduced renal inflammation and IL-1b levels in both the serum and kidneys. SMS-E inhibited both in vitro XO activity and urate uptake in URAT1-expressing oocytes. Using UPLC, 25 ginsenosides were identified, all of which were present in higher levels in SMS-E than in SMS-W. Conclusion: SMS-E exhibited antihyperuricemic effects by regulating XO activity and renal urate transporters, providing the first evidence of its applicability in the treatment of hyperuricemia and gout.

Analysis on electrical and thermal characteristics of MI-SS racetrack coil under conduction cooling and external magnetic field

  • Chae, Yoon Seok;Kim, Ji Hyung;Quach, Huu Luong;Lee, Sung Hoon;Kim, Ho Min
    • Progress in Superconductivity and Cryogenics
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    • v.23 no.4
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    • pp.61-69
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    • 2021
  • This paper presents the analysis and experiment results on the electrical and thermal characteristics of metal insulation (MI) REBCO racetrack coil, which was wound with stainless steel (SS) tape between turn-to-turn layers, under rotating magnetic field and conduction cooling system. Although the field windings of superconducting rotating machine are designed to operate on a direct current, they may be subjected to external magnetic field due to the unsynchronized armature windings during electrical or mechanical load fluctuations. The field windings show the voltage and magnetic field fluctuations and the critical current reduction when they are exposed to an external magnetic field. Moreover, the cryogenic cooling conditions are also identified as the factors that affect the electrical and thermal characteristics of the HTS coil because the characteristic resistance changes according to the cryogenic cooling conditions. Therefore, it is necessary to investigate the effect of external magnetic field on the electrical and thermal characteristics of MI-SS racetrack coil for further development reliable HTS field windings of superconducting rotating machine. First, the major components of the experiment test (i.e., HTS racetrack coil construction, armature winding of 75 kW class induction motor, and conduction cooling system) were fabricated and assembled. Then, the MI racetrack coil was performed under liquid nitrogen bath and conduction cooling conditions to estimate the key parameters (i.e., critical current, time constant, and characteristic resistance) for the test coil in the steady state operation. Further, the test coil was charged to the target value under conduction cooling of 35 K then exposed to the rotating magnetic field, which was generated by three phrase armature windings of 75 kW class induction motor, to investigate the electrical and thermal characteristics during the transient state.

Effect of cryopreservation of ginseng (Panax ginseng C.A. Meyer) seeds on redox ratio of ascorbate and glutathione (인삼종자 초저온보존 후 Ascorbate 및 Glutathione의 산화환원 변화)

  • Baek, Hyung-Jin;Lee, Young-yi;Yoon, Mun-Seop;Song, Jae-young;Balaraju, Kotnala
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2019.04a
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    • pp.81-81
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    • 2019
  • Ginseng seeds are one of short-lived seeds species which loose their viability easily in the condition of conventional storage. Cryopreservation using liquid nitrogen (LN) has been recommended as a alternative storage for this kind of germplasm short lived or dessiccation-sensitive. This study was performed to find out whether cryopreservation could affect physiological change such as enzyme activity induced by reactive oxygen species. In this work, the redox ratio of ascorbate and glutathione were examined onto ginseng seedlings before and after LN storage of seeds for 1 day using spectrophotometer method. Reduced ascorbate (ASA) was increased while oxidized ascorbate (DHA) was decreased slightly for both after 1d-LN storage. And for glutathione also, reduced form (GSH) was increased while oxidized form (GSSG) was decreased slightly for both after 1d-LN storage. Consequently total phenol compound and ion leakage after LN storage showed no significant differences. Additionally root growth from the seeds after LN storage was not affected by ultra low temperature. From the above results, we may suggest that cryopreservation could be recommended for storage tool of ginseng seeds even with low viability also and expected to make slower seed aging process during preservation period through further study.

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Ammonium Acetate Supplement Strategy for Enhancement of Chaetominine Production in Liquid Culture of Marine-Derived Aspergillus fumigatus CY018

  • Liu, Chang-Qing;Wei, Xing-Chen;An, Fa-Liang;Lu, Yan-Hua
    • Journal of Microbiology and Biotechnology
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    • v.29 no.4
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    • pp.587-595
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    • 2019
  • Pharmacological research on (CHA), a marine-derived quinazolinone alkaloid with significant cytotoxic activity, is restricted by low yields and is a problem that needs to be settled urgently. In this work, the selection of additional nitrogen sources and the optimization of additional concentrations and longer fermentation times using ammonium acetate, were investigated. CHA production was optimized to 62.1 mg/l with the addition of 50 mM ammonium acetate at 120 h of the fermentation in the shaker flask. This feeding strategy significantly increased 3-deoxy-arabino-heptulosonate-7-phosphate synthase activity and transcript levels of critical genes (laeA, dahp, and trpC) in the shikimate pathway compared with the non-treatment group. In addition, the selection of the feeding rate (0.01 and $0.03g/l{\cdot}h$) was investigated in a 5-L bioreactor. As a result, CHA production was increased by 57.9 mg/l with a $0.01g/l{\cdot}h$ ammonium acetate feeding rate. This work shows that the strategy of ammonium acetate supplementation had an effective role in improving CHA production by Aspergillus fumigatus CY018. It also shows that this strategy could serve as an important example of large-scale fermentation of a marine fungus in submerged culture.

Contact resistance increment of no-insulation REBCO magnet during a quench

  • Im, Chaemin;Cho, Mincheol;Bang, Jeseok;Kim, Jaemin;Hahn, Seungyong
    • Progress in Superconductivity and Cryogenics
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    • v.21 no.1
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    • pp.31-35
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    • 2019
  • The lumped-parameter circuit model for a no- insulation (NI) high temperature superconductor (HTS) magnet has been well understood after many experimental and analytic studies over a decade. It successfully explains the non-linear charging behaviors of NI magnets. Yet, recently, multiple groups reported that the post-quench electromechanical behaviors of an NI HTS magnet may not be well explained by the lumped circuit model. The characteristic resistance of an NI magnet is one of the key parameters to characterize the so-called "NI behaviors" of an NI magnet and recently a few groups reported a potential that the characteristic resistance of an NI magnet may substantially vary during a quench. This paper deals with this issue, the increment of contact resistance of the no-insulation (NI) REBCO magnet during a quench and its impact on the post-quench behaviors. A 7 T 78 mm NI REBCO magnet that was previously built by the MIT Francis Bitter Magnet Laboratory was chosen for our simulation to investigate the increment of contact resistance to better duplicate the post-quench coil voltages in the simulation. The simulation results showed that using the contact resistance value measured in the liquid nitrogen test, the magnitude of the current through the coil must be much greater than the critical current. This indicates that the value of the contact resistance should increase sharply after the quench occurs, depending on the lumped circuit model.

Cryopreservation of Citrus limon (L.) Burm. F Shoot Tips Using a Droplet-vitrification Method

  • Yi, Jung-Yoon;Balaraju, Kotnala;Baek, Hyung-Jin;Yoon, Mun-Seop;Kim, Haeng-Hoon;Lee, Young-Yi
    • Korean Journal of Plant Resources
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    • v.31 no.6
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    • pp.684-694
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    • 2018
  • This study describes the successful establishment of a cryopreservation protocol for Citrus limon cultivars: 'Frost Eureka limon' and 'Cook Eureka limon', using a droplet-vitrification method. The shoot tips that were excised from in vitro grown seedlings of the two cultivars were preserved in liquid nitrogen (LN) and successfully regenerated into whole plants. Excised shoot tips were pre-cultured for 1 or 2 days in 0.3 M and 0.5 M sucrose solutions at $25^{\circ}C$ and incubated in a loading solution (LS) composed of 17.5% glycerol + 17.5% sucrose in Murashige and Skoog (MS) medium for 40 min at $25^{\circ}C$. Prior to direct immersion in LN for 1 h, the shoot tips were dehydrated with plant vitrification solution 2 (PVS2) at $0^{\circ}C$ or PVS3 at $25^{\circ}C$. The frozen shoot tips were re-warmed and unloaded with 1.2 M sucrose in $\text\tiny{^1/_2}$ MS for 30 min at $25^{\circ}C$. Shoot tips were post-cultured overnight on survival medium and then micrografted onto 'trifoliate orange' (Poncirus trifoliate (L.) Raf. seedling rootstocks for recovery and to produce whole plants. The highest regrowth rates were 53.5% and 50.3% for cryopreserved shoot tips of 'Frost Eureka limon' and 'Cook Eureka limon', respectively, when pre-cultured in 0.3 M and 0.5 M sucrose concentrations in a sequencing manner, with LS and treated with PVS2 for 60 min at $0^{\circ}C$. We also investigated whether the ammonium ion concentration on post-culture medium affected the viability of the cryopreserved Citrus shoot tips. The viability of cooled samples, following culturing on woody plant media (WPM) containing $\text\tiny{^1/_4}$ ammonium nitrate overnight before micrografting, was the highest (70.3%) in 'Frost Eureka limon'. The study described here is a cost-effective and safe method to conserve Citrus fruit cultivars, for the improvement and large-scale multiplication of fruit plants and for breeding disease resistance.

Preliminary Studies for Efficient Treatment of Wastewater Milking Parlor in Livestock Farm (젖소 착유세정폐수의 효율적인 정화처리를 위한 기초연구)

  • Jang, Young Ho;Lee, Soo Moon;Kim, Woong Su;Kang, Jin Young
    • Journal of Korean Society on Water Environment
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    • v.36 no.6
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    • pp.500-507
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    • 2020
  • This study examined the wastewater at a livestock farm, and found that the dairy wastewater from the milking parlor had a lower concentration than the piggery wastewater, and that it was produced at a rate under 1.3 ㎥/day in a single farmhouse. The amount of dairy wastewater was determined based on the performance of the milking machine, the maintenance method of the milking parlor, and the amount of milk production allocated for each farmhouse, not by the area. The results confirmed that both dairy wastewater treatment processes, specifically those using Hanged Bio-Compactor (HBC) and Sequencing Batch Reactor (SBR), can fully satisfy the water quality standards of discharge. The dairy wastewater has a lower amount and concentration than piggery wastewater, meaning it is less valuable as liquid fertilizer, but it can be easily degraded using the conventional activated sludge process in a public sewage treatment plant. Therefore, discharging the dairy wastewater after individual treatment was expected to be a more reasonable method than consigning it to the centralized wastewater treatment plant. The effluent after the SBR process showed a lower degree of color than the HBC effluent, which was attributed to biological adsorption. In the case of the milking parlor in the livestock farm, the concentrations of the effluents obtained after HBC and SBR treatments both satisfied water quality standards for the discharge of public livestock wastewater treatment plants at 99% confidence intervals, and the concentrations of total nitrogen and phosphorous in untreated wastewater were even lower than the water quality standards of discharge. Therefore, we need to discuss strengthening the water quality standards to reduce environmental pollution.