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The Effects of Polyampholyte on Vitrification Process for cryopreservation of Bovine Oviduct Epithelial Cell (Polyampholyte가 소난관상피세포의 초자화 동결방법에 미치는 영향)

  • Kim, Sung Woo;Lee, Jae-Yeong;Kim, Chan-Lan;Yu, Yeonhee;Lee, Sung Soo;Ko, Yeoung-Gyu
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.21 no.6
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    • pp.527-535
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    • 2020
  • The purpose of this study was to establish a simple vitrification protocols to preserve animal cell lines derived from tissues of livestock that could be recultured. Bovine oviduct epithelial cells (BOEC) were used for the vitrification process using a 0.25 ml straw to increase cryopreservation efficiency. BOEC was cultured from the oviduct of 3.5-day estrus state, and the commercially available polyampholyte StemCell KeepTM was used as a cryoprotective agent. Using different concentrations, the viability rates of BOEC in 5, 10, 25, 50, 75, and 100% in freezing media were investigated. Survivability was determined using a differential staining technique using a trypan blue test and a CYTO-13/PI staining protocol. The viability rates of BOEC in the trypan blue test were 5.6±11.8, 12.5±7.2, 53.0±2.7, 85.1±6.9, 79.8±0.6, and 60.7±6.7% with a respective concentration of StemCell KeepTM. The viability rates in CYTO-13/PI staining were 4.6±2.5, 30.8±12.1, 58.4±2.5, 85.5±1.2, 79.8±0.6, and 71.2±1.2%, respectively. These results indicate that BOEC could be preserved with StemCell KeepTM without toxicity in a 0.25-ml straw. The optimal concentration of vitrification solution with StemCell KeepTM was determined to be 50% and can be considered as a proper preservation method for cryobanking.

A Study on Status of Utilization and The Related Factors of Primary Medical Care in a Rural Area (일부 농촌지역의 일차의료이용실태와 그 관련요인에 관한 연구)

  • Wie, Cha-Hyung
    • Journal of agricultural medicine and community health
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    • v.20 no.2
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    • pp.157-168
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    • 1995
  • This study was carried out, through analyzing the annual reports(year of 1973-1993) on health status of Su Dong-Myun, and specific survey data of 332 households(Su Dong-Myun 209, Byul Nae-Myun 123), located in Nam Yang Ju-Si, Kyung Gi-Do, from July 20 to July 31, 1995, to find out more effective means for primary medical care in a rural area. The results were as fellows : 1. Number of population in Su Dong-Myun was 5,419 in 1973, 4,591(the lowest) in 1987 and 5,707 in 1995. In the composition rate of population, "0-14" of age group showed markedly decreasing tendency from 43.1% in 1975, to 19.1% in 1995, however "65 and over" markedly in creasing tendency form 5.3% in 1975 to 9.8% in 1995. 2. Annual utilization rate per 1,000 inhabitants in Su Dong-Myun showed markedly increasing tendency from 1973 to 1977 such as 343 in 1973, 540 in 1975, 900 in 1977. However, since 1979, the rate showed rapidly decreasing tendency, such as 846 in 1979, 519 in 1985, 190 in 1991 and 1993. 3. The morbid household rate per year was 53.6% of respondents and the rate per 15 days was 48.2%. In disease classification rate of morbid household per year, Arthralgia & Neuralgia was the highest rate(33.9%) and gastro-intestinal disorder(19.3%), Cough(11,9%), Hypertension(7.8%), Accident(3.2%) in next order. 4. In the utilizing facilities for Primary Medical Care, Medical facilities was showed the highest rate(58.1% of respondents) and Pharmacy and Drug Shp(33.1%), Tradition Method(4.0%) in next order. In the Medical facilities, General private clinic was showed the highest rate(34.3%) and specific private Clinic(22.3%), Hospital(19.0%), Health (Sub)center(16.3%), Nurse practitioner (3.3%), Oriental hospital and clinic(2.7%) in next order. 5. Experience rate, utilizing health subcenter was 51.8% of the respondents, and it was 55.0% in Su Dong-Myun and 46.3% in Byul Nae-Myun. In utilization times of health subcenter, times-rate showed next orders such as 1-2 times/6months(31.6%), 1-2 times/year (22.1%), 1-2 times/months(19.2%), 1-2 times/3months(15.6%). 6. In objectives, visiting Health Subcenter, Medical Care was the highest rate(59.8% of the respondents) and health control(23.3%) was in next order. In Medical Care, Primary Care by general physician was higher rate(51.1%) almost all. In the Health control, Immunization too was high rate(18.0%) in health control activities. 7. The reasons rate, utilizing health subcenter showed next order, such as distance to Medical facilities(33.0% of the respondents), Medical Cost(28.1%), Simple process of consultation (10.8%), Effectiveness of cure(7.6%), Function of primary medical care(7.0%) and Attitude of physician(6.5%). 8. In the affecting factors to utilization of primary medical facilities, medical needs was showed the highest rate(29.5% of the respondents) and medical cost(15.4%), distance to medical facilities(14.2%), traffic vehicle(14.2%) and farm work(6.9%) in next order. 9. In the priority between 'daily farm work,' and 'primary medical care', only 46.4% of respondents answered that primary health care is more important than the daily farm work The 22.6% of respondents answered 'daily farm work', and the 12.3% answered 'the equal of the both'. 10. In the criterion of medical facilities choice, medical knowledge and technical quality was showed the highest rate(56.3%), distance or time to medical facilities(10.9%), sincerity and kindness of physician(9.4%), medical cost(8.7%) and traffic vehicle(6.5%) in next order 11. In the advise for improvement of health subcenter function, the 36.1% of respondents answered that 'enforcement of medical personnel and equipment' was required, and then 'improved medical technology'(25.5%), 'good attitude of physician'(14.9%), 'improved medical system'(13.3%), 'enforced drug'(6.7%) in next order. 12. The study on affecting factors to utilization of primary medical facilities was very difficult subject to systematize the analyzed results, due to a prejudice of protocol planner, surveyer and respondent, and variety and overlapping of subject matter.

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NORMOBARIC OXYGEN($O_2$) ADMINISTRATION EFFECT ON ATTENTION AND MEMORY FUNCTION IN TEENAGE ADOLESCENTS (10대 청소년의 주의력과 기억능력에 미치는 정상기압 산소흡입 효과)

  • Kim, Byung-Hyo;Kim, Young-Mi;Cho, Soo-Churl;Kim, Boong-Nyun
    • Journal of the Korean Academy of Child and Adolescent Psychiatry
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    • v.13 no.1
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    • pp.76-84
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    • 2002
  • Objectives:This study was conducted to investigate the effect of oxygen on attention and memory functions in healthy adolescents. Methods:The participant subjects were recruited from local advertisement. All subjects are students attending ordinary middle and high school. Their degree of achievement was average or below average. Before the study, its nature and purpose were fully explained to the patients and their parents, and a written informed consent was obtained from each child's parent and a written assent from each child for entire the procedure. The Ethics Committee and Clinical Research Committee of Gyeongsang National University Hospital approved the protocol. For baseline assessment, all subjects received tests for attention and memory. All tests were conducted by a certified psychologist. Stroop test, continuous performance test and trail making test A and B were used for evaluation of attention. As memory tests, we used memory assessment scale(MAS), standardized memory assessment tools. Ten to fourteen days after initial assessments, same tests was applied to the same subjects after prior 5 minute oxygen inhalation. Results:1) Attention test:Improved performances in trail making part B, and stroop test were found in normobaric oxygen inhalation group compared to air inhalation group. Improved reaction time in those tests seemed to reflect the enhanced executive prefrontal activity. 2) Memory test:More words and digits memorization were found in short-term memory subscale score in MAS in oxygen inhalation group compared to air inhalation group. This finding suggested the improved working memory function after oxygen inhalation. Conclusion:Though interpreted cautiously, these results suggested that normobaric oxygen inhalation could enhance executive function and working memory of prefrontal lobe. Further study, however, should be performed to investigate the mechanism of effects of oxygen on cognitive enhancement.

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A study on the exemption of liability of air carriers (항공운송인의 손해배상책임 면제에 관한 법적 고찰)

  • So, Jae-Seon;Lee, Chang-Kyu
    • The Korean Journal of Air & Space Law and Policy
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    • v.30 no.1
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    • pp.95-116
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    • 2015
  • Air transport agreement can be divided into air passenger contract of carriage and aviation also of the contract of carriage. And air carriers for damages greater (1) cause reason, of (2) limit reason, (3) exemption reason. Exemption reason for the extinction of the liability for damages in our Commercial Code, the Convention and domestic law are mixed. Convention on the Commercial Code and air transport, air transport people, if it is proved and that it has taken all the measures that are needed for the prevention of damage to overdue damage of passengers, liability is waived. So what was to achieve the requirements of all the actions that are reasonably necessary in any case is a problem. Amendment has the feature that the treaty for the International Air Transport reflect in accordance with the domestic situation, while being struck by international standards encompassing land, sea and air transport, even on the system. However, Commercial Code while mainly reflect the Montreal Convention governing air carrier's liability issues on the contract of carriage, a problem which the Convention had also began to occur together. So the problem due to accept the treaty to fit the domestic situation occurs. There is a need for analysis of all of the actions that are "reasonably necessary, which is defined in the Commercial Code. If there is no claim within Value Date rotor two years to air carriers on the court for the damage caused by air transport, the responsibility of air carriers disappear, sued the period of such two years, what kind of meaning on domestic law extension and stop to be whether it is interpreted, it should be determined to do their aggressive measures for the reasonable care and accident prevention.

Sow Transfer of Cultured Freezing Embryos by Open Pulled Straw(OPS) Methods : Preliminary Results (Open Pulled Straw(OPS) 방법에 의한 체외배양 동결수정란의 경산돈 이식 : 예비실험 결과)

  • Kim, I.-D.;Ahn, M.-H.;Hur, T.-Y.;Hong, M.-P.;Seok, H.-B.
    • Journal of Embryo Transfer
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    • v.19 no.2
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    • pp.155-163
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    • 2004
  • The aims of this study are 1) to test oocytes and embryos collected from in-vitro to achieving the valuable protocol by culturing, vitrifying and thawing of oocytes/embryos, and 2) to transfer them to recipient, and finally have resulted in pregnancies from recipient females after surgical or nonsurgical transfer. In vitro maturation and fertilization were performed according to Funahashi et al (1994). Glucose-free NCSU 23 supplemented with 5 mM sodium pyruvate, 0.5 mM sodium lactate and 4 mg/ml bovine serum albumin for 2 days at $39^{\circ}C$, and 10% fetal bovine serum albumin was added to the culture medium thereafter. Embryos were treated with 7.5 ${\mu}g/ml$ cytochalasin-B for 30 min, centrifuged at 13,000 rpm for 13 min and then exposed sequentially to an ethylene glycol(EG) vitrification solution, aspirated into OPS, and plunged/thawed into/from liquid nitrogen. In vivo embryos were surgically collected from three dornors after AI for control group. Forty-nine embryos were washed 3 times in mPBS + 10% FBS, followed treatments : cultured, centrifuged, vitrified, recovered and transferred to recipients as in vitro prepared embryos. Three recipients were transferred individually with 100, 100 frozen embryos derived from abattoir and 34 fresh embryos by surgically, and another three recipients were transferred individually with 150, 150 frozen embryos and 100 fresh embryos by nonsurgically, respectively. all recipient sows exhibited delayed returns to estrus. To our knowledge, theses results suggest that required an improved techniques, more vigorous embryos preparation and substitute to gilt with cleaner uterous condition.

Effects of Administration of Oxytocin Antagonist on Implantation and Pregnancy Rates in Patients with Repeated Failure of IVF/ICSI Treatment (체외수정시술의 반복적인 실패 환자에서 옥시토신 길항제 주입이 착상 및 임신에 미치는 영향)

  • Ahn, Jun-Woo;Kim, Chung-Hoon;Kim, So-Ra;Jeon, Gyun-Ho;Kim, Sung-Hoon;Chae, Hee-Dong;Kang, Byung-Moon
    • Clinical and Experimental Reproductive Medicine
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    • v.36 no.4
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    • pp.275-281
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    • 2009
  • Objective: This study was performed to evaluate the effect of oxytocin antagonist on the outcome of IVF/ICSI cycles in infertile patients with repeated failure of IVF/ICSI treatment. Method: Forty patients who had experienced two or more failures of IVF/ICSI treatment without low ovarian reserve, were recruited for this prospective randomized study. All patients received controlled ovarian stimulation (COS) using GnRH antagonist multidose protocol (MDP). For the intervention group, intravenous administration of atosiban (mixed vasopressin $V_{1A}$/oxytocin antagonist) started with a bolus dose 6.75 mg one hour before embryo transfer (ET) and continued at an infusion rate of 18 mg/hour. After ET, administered atosiban was reduced to 6 mg/hour and continued for 2 hours. The main efficacy endpoints were clinical pregnancy rate and implantation rate. Results: Patients' characteristics were comparable in the intervention and control groups. COS parameters and IVF results were also similar. The number of uterine contractions for 3 minutes measured just before ET was significantly lower in the intervention group than control group ($3.5{\pm}1.4$ vs $8.7{\pm}2.2$, p<0.001). While there was no statistically significant difference in the clinical pregnancy rate between control group and intervention group (20.0% and 40.0%, p=0.168), the implantation rate was significantly higher in the intervention group, with 16.9% (11/65) compared with 6.0% (4/67) in the control group (p=0.047). There were no differences in ectopic pregnancy rate and miscarriage rate between the two groups. Conclusion: This study demonstrates that administration of oxytocin antagonist during ET can improve the implantation rate probably by decreasing the frequency of uterine contractions in infertile patients undergoing IVF/ICSI treatment.

tans-Resveratrol Content of Varieties and Growth Period in Peanut (땅콩 품종 및 생육기별 trans-Resveratrol 함량)

  • Lee, Mi-Ja;Cheong, Young-Keun;Kim, Hyung-Soon;Park, Ki-Hun;Doo, Hong-Soo;Suh, Duck-Yong
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.48 no.6
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    • pp.429-433
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    • 2003
  • trans-Resveratrol(3,5,4'-trihydroxystilbene) is phenolic compound present in grapes, wines, and peanuts, has been reported to have health benefits including anticarcinogenic effects, protection against cardiovascular diseases and reduced cancer risk. A simple method for the quantitative extraction of trans-resveratrol from peanut has been developed. Optimal conditions for extraction were investigated. Type of solvent, time, and temperature assayed influenced trans-resveratrol yield. Adequate extraction condition was decided to ethanol/water (80:20v/v) maintained at $25^{\circ}C$ for 45 min. After extraction, the protocol consists of sample preparation using a $\textrm{C}_{18}$ solid-phase extraction (SPE) cartridge after concentrate with rotary evaporator and quantified by reversed phase HPLC using a $\textrm{C}_{18}$ column at 308 nm. Analytical methods for measuring trans-resveratrol in peanut were adapted to isolate, identify, and quantify trans-resveratrol in 11 peanut varieties by HPLC (high-performance liquid chromatography) with UV detector, The 11 peanut varieties content ranged from 0.018 to 1.125 $\mu\textrm{g}/\textrm{g}$ with an average of 0.289 $\mu\textrm{g}/\textrm{g}$. The contents were higher in the seeds with than without testa, regardless of varieties. The trans-resveratrol content was Higher in 110, 130 days after sowing than that of other period.

Pharmacological and Biochemical Characterization of Cells Isolated from Fetal Rat Calvaria (백서태자두개관에서 분리한 세포의 약리학적 및 생화학적 특성에 관한연구)

  • Han, Nam-Soo;Cheong, Dong-Kyun;Mori, Masakazu
    • The Korean Journal of Pharmacology
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    • v.26 no.2
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    • pp.193-207
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    • 1990
  • Transforming growth factor ${\beta}(TGF-{\beta})$ is a multifunctional polypeptide with diverse effects on the proliferation, differentiation and other functions in many cell types. $TGF-{\beta}$ is highly abundant in bone matrix and induces divergent responses in many aspects of bone cell metabolism . Several lines of investigation indicate that matrix-associated $TGF-{\beta}$ is the products of bone cells themselves. However, exact bone cell type reponsible for the production of $TGF-{\beta}$ is still in controversy, The present study was undertaken to determine the cellular origin of matrix-associated $TGF-{\beta}$ and to assess how different bone cells respond to $TGF-{\beta}$. As a prerequisite for this, 5 bone cell populations of distinct phenotype were isolated from fetal calvaria with sequential enzyme digestion protocol and biochemical characterization. Calvarial cell populations released in early stage showed fibroblastic features whereas populations relesed later was enriched with osteoblast-like cell as judged by their acid and alkaline phosphatase activities, cAMP responsiveness to parathyroid hormone, calcitonin and prostaglandin $E_2$ and collagen synthesis rate. By polyacylamide gel and immunoblot analysis of bone and calvarial cell extracts, presence of $TGF-{\beta}$ in bone tissues and production of $TGF-{\beta}$ by bone cells were confirmed again. Subsequent analysis of calvarial cell extracts prepared as individual population revealed that all calvarial cell populations synthesize $TGF-{\beta}$. Exogenously added $TGF-{\beta}$ induced biphasic response upon bone cell proliferation under serum-free condition. In osteoblastic cell populations, it was stimulatory whereas inhibitory in fibroblastic cell populations. In contrast, collagen and noncollagen protein synthesis of all calvarial cell populations were stimulated by $TGF-{\beta}$. Enhancement of protein synthesis was found to be more general rather than specific for collagen synthesis. In addition, effects of $TGF-{\beta}$ on protein synthesis were independent to its effects on cell proliferation. In summary, production of $TGF-{\beta}$ by bone cells and differential actions on various cell populations observed in this study suggest that $TGF-{\beta}$ may play an important role in the regulation of bone metabolism by modulating the specific cellular functions in autocrine and paracrine fashion.

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Analysis of the Amount of Telomeric DNA and Telomerase Activity on Preimplantation Mouse Embryoic Cells (마우스 수정란의 초기 배 발생단계별 Telomeric DNA의 양적 분석과 Telomerase 활성도 분석)

  • Kang M. Y.;Han M. S.;Lee S. C.;Kim J. H.;Sohn S. H.
    • Reproductive and Developmental Biology
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    • v.29 no.1
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    • pp.1-7
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    • 2005
  • Telomeres consisting of (TTAGGG)n tandem repeat DNA sequences and associated proteins are essential for chromosome stability and related with cell senescence, apoptosis and cancer. The telomerase is a ribonucleoprotein which act as a template for the synthesis of telomeric DNA. This study was carried out to identify the distribution of telomeres on mouse chromosomes and also to analyze the amount of telomeres and telomerase activity of mouse embryos at early embryonic stages. Germ cells and early embryos from 1 cell to blastocyst stage were analyzed. The amount of telomeres was analyzed by quantitative fluorescence in situ hybridization technique(Q-FISH) using a human telomeric DNA probe, and telomerase activity was measured by telomeric repeat amplification protocol assay(TRAP). In results, the telomeres on mouse chromosomes were distributed at the ends of all autosomes and sex chromosomes. Although the quantity of telomeres varied among chromosomes, most of chromosomes had higher amount in q-arm telomeres than in p-arm telomeres. The results of Q-FISH indicated that the relative amount of telomeres of mouse embryos in each embryonic stage was approximately the same except the higher amount in blastocysts. Using TRAP assay on mouse embryos, telomerase activity was detected in all preimplantation stages from mature oocytes to blastocysts. Especially the telomerase activity was significantly increased at the morula and blastocyst stage. In conclusion, there may be a close association between the amount of telomeres and telomerase activity in early embryonic stages, and analysis of telomere quantity and telomerase activity on early development will be helpful for the investigation of embryogenesis and embryonic cell differentiation in mice.

The Concept of "Accident" under the Warsaw System (국제항공운송협약상(國際船空運送協約上) 사고(事故)의 개념(槪念))

  • Choi, Jun-Sun
    • The Korean Journal of Air & Space Law and Policy
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    • v.20 no.1
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    • pp.45-85
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    • 2005
  • The purpose of this paper is to examine the concept of "accident" under the Warsaw system including the Warsaw Convention for the Unification of certain Rules for International Carriage by Air of 1929 and the Montreal Convention of 1999. Most leading case on this subject is Air France v. Saks(470 U.S. 392 (1985)). In the Saks case, it was held that the definition of an accident must be applied flexibly, and most courts have adhered to the definition of accident in Saks case, the application of accident has been less than consistent. However, most cases have held that if the event is usual and expected operation of the aircraft, then no accident has occurred. Courts have also held that where the injury results from passenger's own internal reaction to the usual, normal, and expected operations of the aircraft, it is not caused by an accident. As the Warsaw drafters intended to create a system of liability rules that would cover all hazards of air travel, the carrier should liable for the inherent risks of air travel. It is right in that the carrier is in a better position than the passenger to control the risks during air travel. Most US courts have held that carriers are not liable for one passenger's assault on the other passenger. The interactions between passengers are not part of the normal operations of the aircraft and are therefore not covered by the word "accident" under Art 17 of the Warsaw Convention. It is regretful that the Montreal Convention did not attempt to clarify the concepts of accident in itself. In the light of an emerging tendency to hold the air carrier liable for occurrences that do not exactly go to the operation of the aircraft, it is desirable to regulate that the carrier is liable for an "event" instead of an "accident" in accordance with the Guatemala City protocol.

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