• Title/Summary/Keyword: osmotic solution

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Isolation of protoplast from the marine red alga Porphyra pseudolinearis in Korea (한국산 해산 홍조류 긴잎돌김(Porphyra pseudolinearis Ueda)에서의 원형질체 분리)

  • Kim, Young-Dae;Choi, Jae-Seok;Lee, Ju;Son, Yong-Soo;Hong, Yong-Ki
    • Journal of Life Science
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    • v.13 no.4
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    • pp.516-521
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    • 2003
  • Optimal conditions of protoplast isolation from the seaweed Prophyra pseudolinearis have been described. The P. pseudolinearis is one of indigenous and dominant Porphyra species in the East Sea region. Protoplasts have been released by enzymatic treatment of 2% agarase and 2% hemicellulase in 25mM MES buffer, pH 6.0 containing 0.5 M sorbitol. The protoplasts could be fused with neutral red-stained protoplasts of P. okamurae by the addition of polyethylene glycol 8000 solution.

Cryopreservation of in vitro Grown Shoot Tips of Two Freesia hybrida Cultivars by Droplet-vitrification

  • Jinjoo Bae;Jae-Young Song;Ji-Won Han;Ho Cheol Ko;Sung-Hee Nam;Jung-Ro Lee;Ho-sun Lee
    • Korean Journal of Plant Resources
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    • v.36 no.6
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    • pp.562-570
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    • 2023
  • The droplet-vitrification technique for cryopreservation has proven successful across a diverse range of germplasm, ensuring safe and effective long term preservation. In this study, we investigate an effective cryopreservation protocol using the droplet-vitrification technique for shoot tips of Freesia hybrida cultivars 'Sunny Gold' and 'Sweet Lemon'. To determine optimal conditions for Freesia cryopreservation, we employed a carefully selected standard procedure along with additional treatments and alternative solutions. For 'Sunny Gold', the highest regrowth rate of 24% was achieved when shoot tips underwent dehydration with PVS3 solution for 120 minutes before direct immersion in liquid nitrogen (LN) for 1 hour, coupled with a standard protocol involving a two-step preculture with 0.3 M - 0.5 M sucrose, loading with C4 for 40 minutes, and unloading with 0.8 M sucrose for 40 minutes. In the case of 'Sweet Lemon,' regrowth of cryopreserved shoot tips was observed with dehydration treatments, including PVS2 (A3) for 60 minutes and PVS3 (B1) for 60 minutes, as well as longer exposure. The results reflect the distinct sensitivity of shoot tips to chemical toxicity and osmotic stress in these two genotypes. This study provides valuable evidence to consistently enhance the effectiveness of cryopreservation methods for the long-term conservation of Freesia germplasm.

Some Factors Affecting the Protoplast Release from Pleurotus ostreatus (느타리버섯 균사(菌絲)의 원형질체(原形質體) 나출(裸出)에 관한 연구(硏究))

  • Byun, Myung-Ok;Go, Seung-Joo;Park, Yong-Hwan;Shin, Gwan-Chull
    • The Korean Journal of Mycology
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    • v.12 no.1
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    • pp.9-14
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    • 1984
  • Some factors affecting the protoplast release from mycelia of Plurotus ostreatus using commercial lytic enzymes were investigated. The highest yields of the protoplast were obtained from four days old mycelia grown in mushroom complete medium. The solution of 0.8M $MgSO_4$, or KCI showed good results as the osmotic stabilizer for releasing the protoplast. Novozym 234 was the most effective among commercials tested. The concentration of the enzyme and pH of the enzyme solution were optimal at 15mg/ml and $5.5{\sim}6.0$ for the protoplast release, respectively. Mycelial digestion was optimal at about $28^{\circ}C$ and was better in the reciprocal shaking bath (75 oscillations/min) than the stationary culture.

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Functional Integration of Serial Dilution and Capillary Electrophoresis on a PDMS Microchip

  • Chang, Jun-Keun;Heo, Yun-Seok;Hyunwoo Bang;Keunchang Cho;Seok Chung;Chanil Chung;Han, Dong-Chul
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.8 no.4
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    • pp.233-239
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    • 2003
  • For the quantitative analysis of an unknown sample a calibration curve should be obtained, as analytical instruments give relative, rather than absolute measurements. Therefore, researchers should make standard samples with various known concentrations, measure each standard and the unknown sample, and then determine the concentration of the unknown by comparing the measured value to those of the standards. These procedures are tedious and time-consuming. Therefore, we developed a polymer based microfluidic device from polydimethylsiloxane, which integrates serial dilution and capillary electrophoresis functions in a single device. The integrated microchip can provide a one-step analytical tool, and thus replace the complex experimental procedures. Two plastic syringes, one containing a buffer solution and the other a standard solution, were connected to two inlet holes on a microchip, and pushed by a hydrodynamic force. The standard sample is serially diluted to various concentrations through the microfluidic networks. The diluted samples are sequentially introduced through microchannels by electro-osmotic force, and their laser-induced fluorescence signals measured by capillary electrophoresis. We demonstrate the integrated microchip performance by measuring the fluorescence signals of fluorescein at various concentrations. The calibration curve obtained from the electropherograms showed the expected linearity.

A Case of Ischemic Colitis Presenting as Bloody Diarrhea after Glycerin Enema in a Patient on Modified Fasting Therapy (절식요법 중 글리세린 관장 직후 혈성 설사로 발현한 허혈성 대장염 1예)

  • Choi, Hyo-Jeong;Park, Hyun-Gun;Maeng, Tae-Ho;Yoo, Duk-Joo;Kim, Sung-Soo;Chung, Won-Suk
    • Journal of Korean Medicine Rehabilitation
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    • v.23 no.2
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    • pp.185-191
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    • 2013
  • Case of ischemic colitis after enema for bowel cleansing have been rarely reported, but there has been no case report of a patient on modiefied fasting therapy. A 26-year old male patient with obesity admitted Korean medical hospital of Kyung Hee university for losing weight. He is on a special diet called modiefied fasting therapy, only took the fermented herbal drink. At 2nd day, he received an enema for bowel cleansing. A few hours after enema, he had a bloody diarrhea with lower abdominal pain. His colonoscopic and histologic findings presented ischemic colitis. He was advised to fast for two days and couldn't complete his diet program. We suggest 4 possible reasons : Increased intraluminal pressure by enema, vascular spasm caused by room-temperature glycerin solution colder than intraluminal temperature, predisposition to bleeding disturbances by taking selective serotonin reuptake inhibitor(SSRI) for depression history and mucosal injury by osmotic effect of glycerin solution itself. For reducing the risk of bowel cleansing, glycerin enema should be carefully prescribed and practiced concerning the condition of each patient.

The Performance Evaluation of Blended Fertilizers as Draw Solution in Forward Osmosis Desalination (유도용액으로 혼합비료를 사용한 정삼투식 해수담수화에서 혼합비료의 성능평가)

  • Kim, Seung-Geon;Lee, Ho-Won
    • Membrane Journal
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    • v.28 no.1
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    • pp.45-54
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    • 2018
  • This study is to find the optimum draw solution in fertilizer-drawn forward osmosis desalination. Considering osmotic pressure, solubility, and pH, 20 blended fertilizers were screened. Their performance were evaluated in terms of pure water permeate flux, reverse solute flux, and specific reverse solute flux for nitrogen, phosphorus, and potassium. The pure water permeate flux of blended fertilizers including KCl were relatively higher. The reverse solute flux and specific reverse solute flux for nitrogen of blended fertilizers containing ${NO_3}^-$ ion were relatively higher than those of the nitrate ion-free draw solution. Those for phosphorus, and potassium of blended fertilizers including $NH_4H_2PO_4$, and $KNO_3$ were relatively higher than those of the phosphorus-free, and potassium-free draw solution, respectively. The blended fertilizer of $NH_4H_2PO_4$ and KCl contains all of nitrogen, phosphorus and potassium which are essential elements of fertilizer, and has the relatively high water permeation flux and the low reverse solute flux for nitrogen, phosphorus, and potassium. Therefore, it is the most effective draw solution for fertilizer-drawn forward osmosis desalination.

10% Pentastarch Versus 5% Albumin Solution for Volume Expansion Following Cariopulmonary Bypass in Patients Undergoing Open Heart Surgery (개심수술후 혈량 증가를 위한 10% Pentastarch와 5% Albumin 용액의 비교연구)

  • 장병철
    • Journal of Chest Surgery
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    • v.27 no.3
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    • pp.177-186
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    • 1994
  • Pentastarch is a hydroxyethyl starch similar to hetastarch, but lower average molecular weight and fewer hydroxyethyl groups which result in enhanced enzymatic hydrolysis and faster renal elimination.This report was performed to compare the clinical efficacy and safety of 10 % pentastarch[Pentaspan , group I] for plasma volume expansion after open heart surgery with that of 5% albumin[Plasmanate, group II]. There were no statistically significant differences between the group I [n=18] and group II [n:19] in the preoperative parameters [age, sex, body weight] and operative parameters[bypass time, aorta cross clamping time]. During the first 24 hours after arrival of the patient in the surgical intensive care unit, colloid solution [500--1000 ml] was infused to maintain left atrial pressure of more than 8 mmHg, or cardiac index of 2.0 L/min/M2 of more. In results, there were 3 complications of hypotension immediately after infusion of 5 % albumin solution and 2 among the 3 patients were excluded for the study. However there was no complication after infusion of 10 % pentastarch solution. Hemodynamic responses to infusion was similar for both groups, although in group I a greater increase in both left atrial pressure[mean 1.8 versus 0.7 mmHg, p< 0.05] and right atrial pressure [mean 2.2 versus 1.7 mmHg, p < 0.05] was observed during infusion of the first 500 ml. There were no significant differences in any of the measured respiratory parameters[PaO2, intrapulmonary shunt, and effective lung compliance]. Homodilution with colloid significantly reduced hemoglobin [mean 1.2 versus 0.8 gm/dl], and serum protein and albumin level[total protein;4.8$\pm$ 0.5 versus 5.2 $\pm$0.5 gm/dl, p < 0.05: albumin: 3.2 $\pm$0.4 versus 3.6 $\pm$0.6 gm/dl, p < 0.05] by 6:00 AM on 1 day postoperatively, however there were no significant differences on 7 day postoperatively. The mean serum colloid osmotic pressure and osmolarity was similar in both group.There were no abnormal findings of liver function and kidney function in all the patients. There were no significant between-group differences in bleeding time, platelets, prothrombin time, activated partial thromboplastin time and amount of chest tube output measured on 1st and 7th postoperative day. These findings demonstrated that 10% pentastarch is more effective and safe for plasma volume expension than 5 % albumin solution with no adverse effects on coagulation. Also 10 % pentastarch is less expensive than 5 % albumin and it would appeare to be a reasonable first choice for plasma volume expansion.

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Development of hypothermic preservation solution for the human dermal fibroblast using protein hydrolysates (단백질 가수분해 물을 이용한 인간 피부 섬유아세포의 저온 보존액 개발)

  • Byoun, Soon-Hwi;Choe, Tae-Boo
    • KSBB Journal
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    • v.24 no.3
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    • pp.312-320
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    • 2009
  • Stable cell preservation is an essential factor in the regenerative medicine for cell therapies and transplantation of biologic materials. In this study, we studied to provide more stable hypothermic preservation by protection of cell damage during the preservation at $4^{\circ}C$. The result of searching for key components that have excellent efficacy in hypothermic preservation of cells, we have identified the fact that the hypothermic preservation adding protein hydrolysates such as yeast hydrolysate is far superior to others. All protein hydrolysates that are derived from animal, plant and microbe sources have superior efficacy, especially the peptides which have molecular weights under 10 kDa have the best efficacy among the components of protein hydrolysate. The protein hydrolysates prevented the decrease of ATP level in the cells caused by hypothermic environment and they inhibited the generation of ROS. Adding antioxidants and control agents of osmotic pressure were showed to have more superior efficacy in hypothermic preservation. Finally, KUL261 solution (DMEM/F12 1 : 1 medium, yeastolate 1%, $\alpha$-tocopherol $100{\mu}M$, dextran 2.5%), the preservation solution developed in this study, showed the best efficacy in both cell viability and cell growth more than other conventional preservation solutions. In conclusion, the improved hypothermic preservation solution that contains the protein hydrolysates as a key component provide the best preservation efficacy. It provides better efficacy than other preservation solutions and will contribute to both the development of regenerative medicine and global commercialization in this therapeutic field.

Effects of Acetamide and Lactamide on the Viability of Frozen-thawed Mammalian Cells (포유류배양세포 동결보존에 있어 Lactamide의 효과)

  • Kim, Hyun;Cho, Young Moo;Ko, Yeoung-Gyu;Seong, Hwan-Hoo
    • Journal of Life Science
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    • v.24 no.11
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    • pp.1252-1257
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    • 2014
  • While dimethyl sulfoxide (DMSO) is the most commonly used cryoprotectant agent in the cryopreservation of cultured mammalian cells, it has been reported to cause differentiation of some cell lines by DNA methylation and associated histone modifications. To avoid the side effects of DMSO in cryopreservation, other agents might be more appropriate for maintaining the stable differentiation of cultured cell phenotypes through cryopreservation. All cryoprotectants should be highly soluble in water and display low cell toxicity. Cryoprotective agents have been shown to be effective in animal sperm preservation, and eight types of amides were examined in the cryopreservation of cultured mouse endothelial cells. Among the amides examined, acetamide and lactamide were effective cryoprotectants for cultured mammalian cells. The most effective concentration of lactamide, 1.5 M, had an even lower cryoprotective ability than 1M DMSO. Because successful cryopreservation of cultured cells is hampered by osmotic stress, the adequate ionic concentration was determined by diluting phosphate-buffered saline (PBS) in the 1.5M lactamide solution. The most effective concentration was $0.4{\times}PBS$, which minimized osmotic stress during the cryopreservation of cultured cells. As the addition of high molecular weight materials in cryopreservation media improves the viability of cells, the effects of bovine serum albumin (BSA), hydroxyethyl-starch (HES), and dextran were examined. The best combination of lactamide-based media for cryopreservation was found to be 1.5 M lactamide in $0.4{\times}PBS$ with 1% BSA.

The Protoplast Fusion of Aspergillus niger (Aspergillus niger의 원형질체 융합에 관한 연구)

  • Kim, Moo-Sung;Choi, Yong-Keel
    • The Korean Journal of Mycology
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    • v.14 no.2
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    • pp.165-174
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    • 1986
  • Conditions for production, fusion and reversion of protoplasts of Aspergillus niger were investigated, and an attempt was made to enhance fusion frequency. Auxotrophic mutants and morphological mutants were induced by U.V. irradiation $(9.9\;erg/mm^2,\;13min)$ on Aspergillus niger. Maximum yield of protoplasts was obtained from 21 hr cultured mycelia by using 1% driselase in 0.6 M KCl or 0.6 M $NH_4Cl$ as osmotic stabilizer. The optimal temperature for mycelium digestion was $30^{\circ}C$, and the optimal pH was 6.0. Protoplasts produced at different digestion period showed heterogeneity in size and vacuole content. Maximal frequency of protoplasts reversion was obtained on 0.6 M KCl stabilized agar medium at pH 5.0. Reversion frequencies of protoplasts produced for 3 hr and 1 hr mycelial digestion were 8.0% and 15.3%, respectively. The optimal concentration of PEG(m.w. 6000) for protoplast fusion was 30%, and that of $CaCl_2$ was $1{\sim}50\;mM$. The optimal pH and period for the reaction of PEG solution were 8.0 and 10 minutes, respectively. Fusion frequencies between auxotrophic protoplasts produced for 3 hr-mycelial digestion were $0.06{\sim}0.42%$, and those for 1 hr-mycelial digestion were $0.09{\sim}0.54%$.

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