• 제목/요약/키워드: Membrane Integrity Test

검색결과 58건 처리시간 0.251초

체외순환후 fructose-1,6-diphosphatate[FDP]가 적혈구에 미치는 영향 (Effect of Fructose-1,6-diphosphate[FDP] on Red Blood Cells after Extracorporeal Circulation)

  • 이정렬
    • Journal of Chest Surgery
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    • 제25권7호
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    • pp.693-701
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    • 1992
  • Extracorporeal cardiopulmonary bypass[CPB] has been associated with a wide variety of hematologic derangements, including a transient deformation and hemolysis of red blood cells[RBCs], which is supposed to be due to mechanical trauma and/or metabolic alterations. Since membrane integrity is, in part, maintained by energy requiring process, inadequate function of erythrocyte glycolytic pathway, which is inevitalble during CPB, may cause depletion of high energy phosphate pool and result in hemolysis. The authors performed an investigation to assess whether administration of Fructose-l, 6-diphsphate [FDP], which has been known to enhance intracellular glycolytic activities, could counteract erythrocyte hemolytic events caused by CPB. Sixty pateints with cyanotic congenital heart diseases, who underwent open heart surgery under CPB longer than 60 minutes, were randomly divided into two groups depending on whether use of FDP[Group FDP] or not[Group Control]. The age, sex, CPB time, preoperative hemoglobin level, disease entities were all similar[Table 1], and membrane type oxygenators were used in all patients. In Group, FDP, a dose of 250mg/kg body weight of FDP was administered by intravenous dripping every 12 hours from the morning of the operation to postoperative 48 hours, To demonstrate the degree and pattern of hemolysis of erythrocyte, reticulocyte count, indirect /direct bilirubin, haptoglobin, plasma hemoglobin, lactate dehydrogenase were measured every 12 hours from the time of cessation of CPB to 48 hours and RBC morphologic study, osmotic fragility test were done every 24 hours. All parameters revealed less hemolytic in group FDP [Fig. 1~5], though the differences between two groups were not significant, except plasma hemoglobin, lactate dehydrogenase changes. A pattern of sequential changes of plasma hemoglobin, lactate deh-ydrogenase showed the highest level at the time of CPB stop and abrupt decrease in following 24 hours in both groups, and statistically significant differences were demonstrated in group FDP at least for the first 12 hours postoperatively[p<0.05]. The authors conclude that they can expect the benificial effect of FDP on the maintenance of membrane stability of RBC probably by energy enhancement during the shock status of CPB, but FDP could not completely prevent the damaging effect on RBC by cardiopulmonary bypass

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LNG 탱크 방열구조의 슬로싱 충격 응답 해석법에 관한 연구 (A Study on the Sloshing Impact Response Analysis for the Insulation System of Membrane Type LNG Cargo Containment System)

  • 노인식;기민석;이재만;김성찬
    • 한국전산구조공학회:학술대회논문집
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    • 한국전산구조공학회 2011년도 정기 학술대회
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    • pp.531-538
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    • 2011
  • 멤브레인형 LNG 탱크의 구조적 안전성을 확보하기 위해서는 슬로싱에 의한 작용 압력과 구조응답을 정확히 평가할 수 있어야 한다. 탱크 방열구조에 작용하는 슬로싱 충격하중은 매우 불규칙적이며 이로 인한 구조 응답 역시 유탄성 거동을 포함하는 매우 복잡한 물리 현상이기 때문에 최신의 이론적 실험적 접근 방법을 동원하더라도 정확한 평가가 어렵다. 본 연구에서는 실험이나 수치해석으로부터 얻어진 슬로싱 압력 시계열을 이용하여 탱크 방열 구조의 구조응답을 간편하게 해석할 수 있는 방안을 제안하였다. 이 간이 해석법은 기본적인 삼각형 impulse 형태의 충격 압력에 대한 구조응답을 시간영역에서 과도응답해석법으로 계산한 후, 이렇게 구해진 구조물의 삼각형 응답함수를 조합하여 임의 형상의 압력 시계열에 대한 구조 응답을 구하는 방식이다. 여러 가지 예제 해석을 통하여 제안된 해석법의 타당성을 검토하였고, 이를 이용하여 실제 모형실험에서 얻어진 압력시계열을 바탕으로 구조응답을 계산하고 그 결과를 고찰하였다.

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Effects of Taurine on Sperm Characteristics during In vitro Storage of Boar Semen

  • Jang, H.Y.;Kong, H.S.;Park, C.K.;Oh, J.D.;Lee, S.G.;Cheong, H.T.;Kim, J.T.;Lee, S.J.;Yang, B.K.;Lee, H.K.
    • Asian-Australasian Journal of Animal Sciences
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    • 제19권11호
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    • pp.1561-1565
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    • 2006
  • The objective of this study was to investigate the anti-oxidative effects of taurine on sperm characteristics for in vitro storage of boar semen. Semen was randomly divided into 10 groups in conical tubes and treated with different concentrations of taurine (25-100 mM) with or without $250{\mu}M$ $H_2O_2$. The percentage of motile spermatozoa in taurine groups after 6 and 9 h were significantly higher at >94% and 87%, respectively, compared to the control group ($85.1{\pm}0.5$ and $72.4{\pm}0.3$, p<0.05). The sperm motility in taurine with $H_2O_2$ after 6 h incubation was slightly decreased compared to the taurine alone treatment, but after 9 and 12 h incubation % sperm motility dropped sharply in taurine with $H_2O_2$ ($75.3{\pm}0.3$ and $69.6{\pm}2.9$, p<0.05). For 3, 9 and 12 h incubation, sperm viability in the control was lower than in taurine groups, irrespective of taurine concentration. In eosin Y and nigrosin staining (ENS), the sperm survival rates (%) for 6 h incubation were significantly higher in 25 mM ($76.0{\pm}0.6$) and 50 mM taurine groups ($78.0{\pm}0.7$), respectively. Sperm survival rates for 9 and 12 h incubation were higher in taurine groups (${\geq}48%$ in 9 h and ${\geq}42%$ in 12 h) compared to controls ($43.0{\pm}2.1$ and $31.0{\pm}0.6$, respectively). In the hyoosmotic swelling test (HOST), sperm membrane integrity was similar to the results of sperm survival. These experiments indicate that supplementation of taurine to the semen extender can increase the sperm characteristics(motility, viability, survival and membrane integrity).

Effect of Extenders and Temperatures on Sperm Viability and Fertilizing Capacity of Harbin White Boar Semen during Long-term Liquid Storage

  • Zhou, J.B.;Yue, K.Z.;Luo, M.J.;Chang, Z.L.;Liang, H.;Wang, Z.Y.;Tan, J.H.
    • Asian-Australasian Journal of Animal Sciences
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    • 제17권11호
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    • pp.1501-1508
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    • 2004
  • In this study the effect of extenders and temperatures on sperm viability and fertilizing capacity of boar sperm during long-term storage was investigated. Acrosomal integrity, membrane integrity, motility and hypo-osmotic resistance were evaluated by fluorescence and light microscopy. An in vitro fertilization test was performed to assess the fertilizing capacity of stored spermatozoa. The five diluents tested were ranked according to their ability to maintain sperm functional parameters and Zorlesco (ZO) extender with BSA or with PVA instead of BSA produced the best results. Zorlesco extender substituted with PVA (ZO+PVA) was found to maintain motility both at 15 and 20$^{\circ}C$. within 5 days of storage, but the quality of semen stored at 15$^{\circ}C$ decreased thereafter as compared to semen stored at 20$^{\circ}C$ Semen stored at 5$^{\circ}C$ demonstrated rapid loss of motility already within 24 h. Both fertilization and cleavage of semen stored at 20$^{\circ}C$ in ZO substituted with PVA instead of BSA did not change significantly until day 8 of storage. It is therefore concluded that PVA can be used to substitute for BSA and 20$^{\circ}C$ was more suitable than 15$^{\circ}C$ for boar semen storage, and in vitro fertilizing capacity of spermatozoa was maintained for at least 8 days in ZO+PVA at 20$^{\circ}C$.

공기압식 외력부가방법을 이용한 구조강도 시험기법 연구 (The Study on Structural Strength Test Technique by Using Compressed Air Type Loading Method)

  • 김종환;이기범;김호연
    • 한국항공우주학회지
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    • 제38권4호
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    • pp.376-381
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    • 2010
  • 일반적으로 기체 구조물 개발시 운용조건에서의 구조설계 해석 검증 및 구조강도확인을 위해서 구조강도시험이 수행된다. 본 논문에서는 구조시험 하중부가방법 중 응력집중에 의해 파손되기 쉬운 재료를 기체 지지구조로 사용하고자 할 때 구조물 표면에 공기압을 이용하여 국부적 응력집중을 피하면서 분포하중을 부가할 수 있는 새로운 외력부가장치의 개발에 대하여 기술하였다. 본 장치는 부드러운 재질의 하중전달용 매개체를 이용하여 구조물 표면에 손상을 주지 않고 외력을 전달할 수 있으며, 임의형상 구조물에 대하여 쉽게 응용이 가능하다. 본 장치를 이용하여 평판구조물과 곡면구조물에 대하여 외력부가구조강도시험을 성공적으로 수행함으로써 적용시험기법의 타당성을 입증하였다.

점안액 보존제 성분 Benzalkonium Chloride에 의해 유도된 각막상피세포의 세포고사 유도 (Effects of Benzalkonium Chloride on the growth and survival of Human corneal epithelial cells)

  • 김재민;이석주;서은선
    • 한국안광학회지
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    • 제7권2호
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    • pp.189-195
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    • 2002
  • 점안액 보존제 성분으로 이용되는 Benzalkonium Chloride(BAC)가 배양 각막 상피세포에 미치는 영향을 조사하기 위해 본 연구를 시행하였다. BAC를 세포에 0.0001%~0.01%로 15분 동안 처리하여 24시간 후 회복 효과를 조사하였다. MTT assay를 이용하여 세포 생존율을 산정하고 Hoechst 33342로 염색질 응축을 조사하였다. Fas 발현 여부를 조사하기 위해 western blot과 immunocytochemistry를 이용하였으며 DNA fragmentation은 agarose gel을 이용한 전기영동을 시행하였다. BAC를 0.005% 이상 처리하면 세포괴사가 일어나는 반면에 저농도의 BAC는 세포고사를 유도하는 것으로 나타났으며 BAC로 Fas 발현이 유도될 수 있다는 것을 보여주고 있다. 본 연구를 통해 점안액의 대표적인 보존제인 BAC의 부작용에 대한 각막세포 차원의 연구가 필요하며 세포독성이 없는 새로운 보존제의 개발이 필요한 것으로 나타났다.

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Progesterone 및 Acetyl-L-Carnitine이 정자의 동결-융해에 미치는 영향 (The Effects on Sperm Parameters and Membrane after Treatment with Progesterone and/or Acetyl-L-Carnitine; Cryopreservation-Thawing)

  • 정병준;김윤진;최형민;전명권;이응수;나오순
    • Clinical and Experimental Reproductive Medicine
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    • 제28권4호
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    • pp.295-300
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    • 2001
  • Objective : To assess the effects of progesterone and acetyl-L-carnitine used after treated with Isolate�� gradient before semen cryopreservation-thawing on sperm parameters and membrane integrity. Material and Methods : From April 2001 to July 2001, ten normal male partner of couples who were visited in vitro fertilization (IVF) clinics. the semens were treated with $Isolate^{(R)}$ gradient before cryopreservation, spermatozoa was incubated with progesterone (1, 5 and $10{\mu}M$), acetyl-L-carnitine (2.5, 5 and $10{\mu}M$), or both (progesterone, $1{\mu}M$; and acetyl-L-carnitine, $5{\mu}M$) for 30 min. Results: There were no differences in sperm parameters and vital stain among isolate only treated group, progesterone (1, 5 and $10{\mu}M$), acetyl-L-carnitine (2.5, 5 and $10{\mu}M$) and both (progesterone, $1{\mu}M$; and acetyl-L-carnitine, $5{\mu}M$). But, in high concentration of acetyl-L-carnitine ($10{\mu}M$) treated group, sperm parameters and vital stain were decreased. The statistical method was used ANOVA (Kruskal-Wallis test) and p value was <0.01. Conclusions : Neither progesterone nor acetyl-L-carnitine show to be protective effect on the cryodamage assessed by sperm parameters and vital stain (eosin-Y stain) in normal sperm. High concentration of acetyl-L-carnitine ($10{\mu}M$), however, was harmful effect on cryoprevention.

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Changes in Acrosin Activity and Membrane Function of Boar Spermatozoa

  • Kim, C.K.;Kim, I.;Y.C. Chung;J.W. Ryu;H.J. Yoon;K. Kang;Kim, I.C.;Lee, J.H.;S.E. Yeon
    • 한국동물번식학회:학술대회논문집
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    • 한국동물번식학회 2001년도 춘계학술발표대회
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    • pp.86-86
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    • 2001
  • The aims of this work were to determine the acrosin activity and to evaluate the structural and functional integrity of AS boar spermatozoa. The acrosin activity of spermatozoa were 5.40, 4.10 and 3.40 mIU/10$^{6}$ sperm in raw, extended and frozen semen respectively , which differed significantly each other (P<0.05). After the raw and extended semen were exposured to cold and thermal shock, the acrosin activities of spermatozoa in the raw semen were 5.39, 5.21 and 5.29 mIU/10$^{6}$ sperm for control (non-shock), cold shock and thermal shock, and those of extended semen were 4.21, 3.98 and 4.00 mIU/10$^{6}$ sperm. This value among treatments did not differ significantly. The acrosin activities of spermatozoa in the extended and stored semen were 3.27, 3.52, 3.46 and 3.23 mIU/10/suup 6/ sperm, while hypo-osmotic test(HOST) values were 56.5%, 64.7%, 66.0% and 56.0%, following 4 days storage at 4$^{\circ}C$, 17$^{\circ}C$ , $25^{\circ}C$ and 37$^{\circ}C$, respectively. The results at 17$^{\circ}C$ and $25^{\circ}C$ appeared to be best compared with the other storage temperatures.

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정자 운동성 및 수명 보존을 위한 최적 배양에 관한 연구 (Optimized Methods to Maintain Motility and Viability in Normozoospermic Males)

  • 유영아;;오신애;방명걸
    • Clinical and Experimental Reproductive Medicine
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    • 제36권1호
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    • pp.45-53
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    • 2009
  • 목 적: 본 연구는 정상 정액을 배양액으로 세척한 후 $4^{\circ}C$, $22^{\circ}C$, $37^{\circ}C$에서 5일 동안 보존하면서 정자의 운동성, 생존성을 관찰하여 정자의 운동성과 수명 유지를 위한 적정 배양 환경을 분석하고자 하였다. 연구방법: 정액검사 시 정상으로 판정된 남성의 정자를 HTALP 배양액으로 세척하여 정자의 최종 농도 $1{\times}10^6/ml$로 각 5 ml을 배양온도 $4^{\circ}C$, $22^{\circ}C$, $37^{\circ}C$에서 5일 동안 배양하였다. 1일, 3일 5일째에 CASA에 의해 운동성을 측정하였고, HOST로 정자막의 온전성을 분석하였으며, CTC pattern으로 수정능획득 상태 분석하여 최적의 배양 환경을 분석하였다. 결 과: 정자의 운동성, 생존성 및 수정능획득이 야기되지 않은 정자는 배양일에 따라 유의하게 감소하였다 (p<0.05). 또한 정자 배양 후 1일에는 정자의 운동성, 생존성 및 정자막의 온전성과 CTC pattern은 온도에 따라 차이가 없었으나, 배양 후 3일과 5일에서는 $22^{\circ}C$에서 배양된 정자가 다른 배양온도에 비해 가장 잘 보존되었다 (p<0.05). 결 론: HTALP로 세척된 정자를 $22^{\circ}C$에서 보존 시 5일까지 정자의 운동성과 수명을 보조생식술에 적합한 수준으로 유지시킬 수 있는 최적 배양 환경으로 제시할 수 있다.

Seminal Plasma Heparin Binding Proteins Improve Semen Quality by Reducing Oxidative Stress during Cryopreservation of Cattle Bull Semen

  • Patel, Maulikkumar;Gandotra, Vinod K.;Cheema, Ranjna S.;Bansal, Amrit K.;Kumar, Ajeet
    • Asian-Australasian Journal of Animal Sciences
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    • 제29권9호
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    • pp.1247-1255
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    • 2016
  • Heparin binding proteins (HBPs) are produced by accessory glands. These are secreted into the seminal fluid, bind to the spermatozoa at the time of ejaculation, favour capacitation, acrosome reaction, and alter the immune system response toward the sperm. The present study was conducted with an objective to assess the effect of purified seminal plasma-HBPs (SP-HBPs) on cross bred cattle bull sperm attributes during two phases of cryopreservation: Pre freezing and freezing-thawing. SP-HBPs were purified from pooled seminal plasma by heparin affinity chromatography. Three doses of SP-HBPs i.e. 10, 20, $40{\mu}g/mLs$ semen were standardized to find out the optimum dose and $20{\mu}g/mLs$ was found to be an optimum dose. Semen as such and treated with SP-HBPs was diluted with sodium citrate-egg yolk diluter and cryopreserved as per the standard protocol. Sperm parameters i.e. motility, viability, Hypo-osmotic swelling test (HOST), acrosome damage, in vitro capacitation and lipid peroxidation were evaluated in SP-HBP treated and untreated (control) semen at both phases of cryopreservation. A considerable variation in percent sperm motility, viability, membrane integrity (HOST), acrosome damage, acrosome reaction and lipid peroxidation was observed at both phases among the bulls irrespective of the treatment. Incubation of neat semen with $20{\mu}g/mL$ SP-HBP before processing for cryopreservation enhanced the average motility, viability, membrane integrity by 7.2%, 1.5%, 7.9%, and 5.6%, 6.6%, 7.4% in pre-frozen and frozen-thawed semen in comparison to control. There was also an average increase of 4.1%/3.9% in in vitro capacitation and acrosome reaction in SP-HBPs-treated frozen-thawed semen as compared to control. However, binding of SP-HBPs to the sperm declined acrosome damage and lipid peroxidation by 1.3%/4.1% and 22.1/$32.7{\mu}M$/$10^9$ spermatozoa in SP-HBP treated pre-frozen/frozen-thawed semen as compared to control, respectively. Significant (p<0.05) effects were observed only in motility, HOST and in vitro acrosome reaction. It can be concluded that treatment of neat semen with SP-HBPs before cryopreservation minimized the cryoinjury by decreasing the generation of reactive oxygen species.