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Prediction of Parthenogenetic Developmental Potential by Polar Body Extrusion and First Cleavage on In Vitro Maturation and Development of Porcine Follicular Oocytes (돼지 난모 세포의 Ethanol 처리에 의한 단위 발생에 있어서 극체 방출란과 분할란 선별에 따른 배발달율 비교)

  • Kim, H.J.;Cho, S.R.;Choe, J.Y.;Choi, S.H.;Han, M.H.;Son, D.S.;Lee, S.S.;Sang, B.D.;Ryu, I.S.;Kim, I.C.;Kim, S.J.;Kim, I.H.;Kim, S.K.
    • Journal of Embryo Transfer
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    • v.22 no.2
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    • pp.121-126
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    • 2007
  • The objective of this study was carried out to examine the selection effects of in vitro matured porcine follicular oocytes with polar body extrusion and early cleavage as non-invasive marker to know the developmental competence in advance. The porcine oocytes matured for 48 h were examined the polar body extrusion. The examined oocytes were matured for additional $16{\sim}18h$ and activated with 7% ethanol and cultured in $5{\mu}g/ml$ cytochalasin B for 5 h for diploid formation. The treated oocytes were cultured and examined the cleavage after 48 h and continued culturing for 5 days. The oocytes of 21.9% were discarded in morphological selection and 32.1% oocytes were discarded by failure of first polar body extrusion. The selected oocytes were matured and activated and then after 48 h the cleavage rates were examined. In morphologically selected oocytes, 15.8% oocytes were not cleaved and 52.6% oocytes were normally cleaved and 31.6% oocytes were hyper-cleaved over 8-cell stage. However in the first polar body extruded oocytes, 7.1% oocytes were not cleaved and 73.1% oocytes were normally cleaved and 19.8% oocytes were hyper-cleaved. The morphologically selected embryos that not cleavage-selected were developed in 16.7% up to blastocyst and the morphologically selected and cleavage-selected embryos were developed in 31.7%. The polar body extruded oocytes that were not carried out cleavage selection were developed in 39.0% and the polar body extruded and cleavage-selected embryos were developed 49.0%. The first cleavage timing was examined with 12 h interval after activation. In $0{\sim}12,\;12{\sim}24,\;24{\sim}36,\;and\;36{\sim}48h$ intervals, 4.1%, 68.6%, 19.1%, and 2.3% oocytes were cleaved and 5.9% oocytes were not cleaved until 48 after activation. The cleaved oocytes in each interval were cultured and developed upto blastocyst with 0, 39.1, 9.5, and 0%, respectively. This results suggests that polar body extruded and cleaved at $12{\sim}36h$ embryo has higher developmental potential than the others.

A Study on Influence of Foodservice Managers' Emotional Intelligence on Job Attitude and Organizational Performance (급식관리자의 개인적 감성지능이 직무태도 및 조직성과에 미치는 영향)

  • Jung, Hyun-Young;Kim, Hyun-Ah
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.39 no.12
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    • pp.1880-1892
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    • 2010
  • The purposes of this study were to: a) provide evidence concerning the effects of emotional intelligence on job outcomes, b) examine the impacts of emotional intelligence on employee-related variables such as 'job satisfaction', 'organizational commitment', 'organizational performance', and 'turnover intention' c) identify the conceptual framework underlying emotional intelligence. A survey was conducted to collect data from foodservice managers (N=231). Statistical analyses were completed using SPSS Win (16.0) for descriptive analysis, reliability analysis, factor analysis, t-test, correlation analysis, cluster analysis and AMOS (16.0) for confirmatory factor analysis and structural equation modeling. The concept of emotional intelligence (EI) has been on the radar screens of many leaders and managers over the last several decades. The emotional intelligence is generally accepted to be a combination of emotional and interpersonal competencies that influence behavior, thinking and interaction with others. The main results of this study were as follows. The four EI (Emotional Intelligence) dimensions correlated significantly with age. The means of job satisfaction score were above the midpoint (3.04 point) scale. The organizational commitment score was above the midpoint (3.41 point) scale and was higher at 'loyalty' factor than 'commitment' factor. The means of organizational performance score were above the midpoint (3.34) scale. The correlations among the four EI (emotional intelligence) factors were significant with job satisfaction; organizational commitment, organizational performance and turnover intention. The test of hypothesis using structural equation modeling found that emotional intelligence produced positive effects on job attitude and job performance. Emotional intelligence enhanced organizational commitment, and in turn, managers' attitude produced positive effects on organizational performance; emotional intelligence also had a direct impact on organizational performance. This study has identified the effect of emotional intelligence on organizational performance and attitudes toward one's job.

INFLUENCES OF DRY METHODS OF RETROCAVITY ON THE APICAL SEAL (치근단 역충전와동의 건조방법이 폐쇄성에 미치는 영향)

  • Lee, Jung-Tae;Kim, Sung-Kyo
    • Restorative Dentistry and Endodontics
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    • v.24 no.1
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    • pp.166-179
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    • 1999
  • Apical sealing is essential for the success of surgical endodontic treatment. Root-end cavity is apt to be contaminated with moisture or blood, and is not always easy to be dried completely. The purpose of this study was to evaluate the influence of dry methods of retrocavity on the apical seal in endodontic surgery. Apical seal was investigated through the evaluation of apical leakage and adaptation of filling material over the cavity wall. To investigate the influence of various dry methods on the apical leakage, 125 palatal roots of extracted human maxillary molar teeth were used. The clinical crown of each tooth was removed at 10 mm from the root apex using a slow-speed diamond saw and water spray. Root canals of the all the specimens were prepared with step-back technique and filled with gutta-percha by lateral condensation method. After removing of the coronal 2 mm of filling material, the access cavities were closed with Cavit$^{(R)}$. Two coats of nail polish were applied to the external surface of each root. Apical three millimeters of each root was resected perpendicular to the long axis of the root with a diamond saw. Class I retrograde cavities were prepared with ultrasonic instruments. Retrocavities were washed with physiologic saline solution and dried with various methods or contaminated with human blood. Retrocavities were filled either with IRM, Super EBA or composite resin. All the specimens were immersed in 2% methylene blue solution for 7 days in an incubator at $37^{\circ}C$. The teeth were dissolved in 14 ml of 35% nitric acid solution and the dye present within the root canal system was returned to solution. The leakage of dye was quantitatively measured via spectrophotometric method. The obtained data were analysed statistically using one-way ANOVA and Duncan's Multiple Range Test. To evaluate the influence of various dry methods on the adaptation of filling material over the cavity wall, 12 palatal roots of extracted human maxillary molar teeth were used. After all the roots were prepared and filled, and retrograde cavities were made and filled as above, roots were sectioned longitudinally. Filling-dentin interface of cut surfaces were examined by scanning electron microscope. The results were as follows: 1. Cavities dried with paper point or compressed air showed less leakage than those dried with cotton pellet in Super EBA filled cavity (p<0.05). However, there was no difference between paper point- and compressed air-dried cavities. 2. When cavities were dried with compressed air, dentin-bonded composite resin-filled cavities showed less apical leakage than IRM- or Super EBA-filled ones (p<0.05). 3. Regardless of the filling material, cavities contaminated with human blood showed significantly more apical leakage than those dried with compressed air after saline irrigation (p<0.05). 4. Outer half of the cavity showed larger dentin-filling interface gap than inner half did when cavities were filled with IRM or Super EBA. 5. In all the filling material groups, cavities contaminated with blood or dried with cotton pellets only showed larger defects at the base of the cavity than ones dried with paper points or compressed air.

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Steroid Effect on the Brain Protection During OPen Heart Surgery Using Hypothermic Circulatory Arrest in the Rabbit Cardiopulmonary bypass Model (저체온순환정지법을 이용한 개심술시 스테로이드의 뇌보호 효과 - 토끼를 이용한 심폐바이패스 실험모델에서 -)

  • Kim, Won-Gon;Lim, Cheong;Moon, Hyun-Jong;Chun, Eui-Kyung;Chi, Je-Geun;Won, Tae-Hee;Lee, Young-Tak;Chee, Hyun-Keun;Kim, Jun-Woo
    • Journal of Chest Surgery
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    • v.30 no.5
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    • pp.471-478
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    • 1997
  • Introduction: The use of rabbits as a cardiopulmonary bypass(CPB) animal model is extremely dif%cult mainly due to technical problems. On the other hand, deep hypothermic circulatory arrest(CA) is used to facilitate surgical repair in a variety of cardiac diseases. Although steroids are generally known to be effective in the treatment of cerebral edema, the protective effects of steroids on the brain during CA are not conclusively established. Objectives of this study are twofold: the establishment of CPB technique in rabbits and the evaluation of preventive effect of steroid on the development of brain edema during CA. Material '||'&'||' Methods: Fifteen New Zealan white rabbits(average body weight 3.5kg) were divided into three experimental groups; control CA group(n=5), CA with Trendelenberg position group(n=5), and CA with Trendelenberg position + steroid(methylprednisolone 30 mglkg) administration group(n=5). After anesthetic induction and tracheostomy, a median sternotomy was performed. An aortic cannula(3.3mm) and a venous ncannula(14 Fr) were inserted, respectively in the ascending aorta and the right atrium. The CPB circuit consisted of a roller pump and a bubble oxygenator. Priming volume of the circuit was approximately 450m1 with 120" 150ml of blood. CPB was initiated at a flow rate of 80~85ml/kg/min, Ten min after the start of CPB, CA was established with duration of 40min at $20^{\circ}C$ of rectal temperature. After CA, CPB was restarted with 20min period of rewarming. Ten min after weaning, the animal was sacrif;cod. One-to-2g portions of the following tissues were rapidly d:ssected and water contents were examined and compared among gr ups: brain, cervical spinal cord, kidney, duodenum, lung, heart, liver, spleen, pancreas. stomach. Statistical significances were analyzed by Kruskal-Wallis nonparametric test. Results: CPB with CA was successfully performed in all cases. Flow rate of 60-100 mlfkgfmin was able to be maintained throughout CPB. During CPB, no significant metabolic acidosis was detected and aortic pressure ranged between 35-55 mmHg. After weaning from CPB, all hearts resumed normal beating spontaneously. There were no statistically significant differences in the water contents of tissues including brain among the three experimental groups. Conclusion: These results indicate (1) CPB can be reliably administered in rabbits if proper technique is used, (2) the effect of steroid on the protection of brain edema related to Trendelenburg position during CA is not established within the scope of this experiment.

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Second allogeneic hematopoietic stem cell transplantation in children to overcome graft failure or relapse after initial transplant (조혈모세포이식 후 생착 실패나 재발한 소아환자에서 2차 이식의 의의)

  • Kim, Dong-Yeon;Kim, Do Kyun;Kim, Soo Young;Kim, Seok Joo;Han, Dong Gyun;Baek, Hee Jo;Kook, Hoon;Hwang, Tai-Ju
    • Clinical and Experimental Pediatrics
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    • v.49 no.12
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    • pp.1329-1339
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    • 2006
  • Purpose : Failure of hematopoietic stem cell transplantation(HSCT) may be encountered in practice because of either relapse of the malignancy or dysfunction of the graft. Second HSCT may be the only option for some patients whose initial HSCT failed. Methods : From May, 1991 to December, 2004, 115 HSCTs were performed at the Pediatric Blood & Marrow Transplantation Center, Chonnam National University. This study was a retrospective analysis of the medical records of 15 patients who received the second HSCT after initial graft. Results : Among eight patients with nonmalignant diseases, two patients underwent the second HSCT because of primary graft failure and five because of late graft rejection. The remaining Fanconi anemia patient was re-transplanted due to development of AML. Two patients died and one experienced primary graft failure, but is still alive. The Kaplan-Meier 5-year overall survival rate was 75 percent and the disease free survival rate was 62.5 percent in nonmalignant diseases. All malignant patients underwent second transplants because of relapses. Four died of relapse and one of treatment-related complications. The Kaplan-Meier 2-year overall and event free survival rate was 28.6 percent each in malignant diseases. Conclusion : Second HSCT for graft dysfunction of nonmalignant disease seems to be feasible and should be considered as a standard practice. The relapse of malignant diseases remains a big obstacle even after the second HSCT, although a small portion of patients might be salvaged. Further investigation of novel therapeutic strategies, as well an the understanding of the biology should be explored.

[ $^1H$ ] MR Spectroscopy of the Normal Human Brains: Comparison between Signa and Echospeed 1.5 T System (정상 뇌의 수소 자기공명분광 소견: 1.5 T Signa와 Echospeed 자기공명영상기기에서의 비교)

  • Kang Young Hye;Lee Yoon Mi;Park Sun Won;Suh Chang Hae;Lim Myung Kwan
    • Investigative Magnetic Resonance Imaging
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    • v.8 no.2
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    • pp.79-85
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    • 2004
  • Purpose : To evaluate the usefulness and reproducibility of $^1H$ MRS in different 1.5 T MR machines with different coils to compare the SNR, scan time and the spectral patterns in different brain regions in normal volunteers. Materials and Methods : Localized $^1H$ MR spectroscopy ($^1H$ MRS) was performed in a total of 10 normal volunteers (age; 20-45 years) with spectral parameters adjusted by the autoprescan routine (PROBE package). In all volunteers, MRS was performed in a three times using conventional MRS (Signa Horizon) with 1 channel coil and upgraded MRS (Echospeed plus with EXCITE) with both 1 channel and 8 channel coil. Using these three different machines and coils, SNRs of the spectra in both phantom and volunteers and (pre)scan time of MRS were compared. Two regions of the human brain (basal ganglia and deep white matter) were examined and relative metabolite ratios (NAA/Cr, Cho/Cr, and mI/Cr ratios) were measured in all volunteers. For all spectra, a STEAM localization sequence with three-pulse CHESS $H_2O$ suppression was used, with the following acquisition parameters: TR=3.0/2.0 sec, TE=30 msec, TM=13.7 msec, SW=2500 Hz, SI=2048 pts, AVG : 64/128, and NEX=2/8 (Signa/Echospeed). Results : The SNR was about over $30\%$ higher in Echospeed machine and time for prescan and scan was almost same in different machines and coils. Reliable spectra were obtained on both MRS systems and there were no significant differences in spectral patterns and relative metabolite ratios in two brain regions (p>0.05). Conclusion : Both conventional and new MRI systems are highly reliable and reproducible for $^1H$ MR spectroscopic examinations in human brains and there are no significant differences in applications for $^1H$ MRS between two different MRI systems.

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A Study on Oriental Medical Diagnosis of Musculoskeletal Disorders using Moire Image (Moire 영상을 이용한 근골격계 질환의 한의학적 진단에 관한 연구)

  • Lee Eun-Kyoung;Yu Seung-Hyun;Lee Su-Kyung;Kang Sung-Ho;Han Jong-Min;Chong Myong-Soo;Chun Eun-Joo;Song Yung-Sun;Lee Ki-Nam
    • Journal of Society of Preventive Korean Medicine
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    • v.4 no.2
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    • pp.72-92
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    • 2000
  • This research has conducted studies on an Oriental medicine-based method of diagnosing of occupational musculoskeletal system diseases. This researcher has searched through existing relevant medical literature. Also, this researcher has worked on a moire topography using moire topography. In this course, this researcher has reached the following conclusion in relation to the possibility of using a moire topography as a diagnosing device of musculoskeletal system diseases under Oriental medicine . 1 The Western medicine outlines its criteria of screening occupational musculoskeletal system diseases as follows A. The occupational musculoskeletal diseases must clearly include one or more of the subjective symptoms characterized by pain, hypoesthesia dysaesthesia, anaesthesia. etc . B, There should be clinically admitted objective observations and diagnosis outlining that the disease concerned shows symptoms such as tenderness, induration. and edema that can appear with occupational musculoskeletal system diseases. dyscinesia should be admitted with the disease concerned, or there should be observations and diagnosis outlining that abnormality exists in electric muscular or nervous diagnosis and examination . C. It should be admitted that prior to the occurrence of symptoms or observations and diagnosis on musculoskeletal system-related diseases, a patient has been engaged in works with conditions requiring improper work posture or work movement. That is, this is an approach whereby they see abnormality in the musculoskeletal system come from material and structural defect, and adjust and control abnormality in the musculoskeletal system and secreta . 2. The Oriental medicines sees that a patient develops the pain of occupational musculoskeletal diseases as he cannot properly activate the flow of his life force and blood thus not only causing formation of lumps in the body and blocking the flow of life force and blood in some parts of the body. Hence, The Oriental medicine focuses on resolving the cause of weakening the flow of life force and blood, instead of taking material approach of correcting structural abnormality Furthermore , Oriental medicine sees that when muscle tension builds up, this presses blood vessels and nerves passing by, triggering circulation dyscrasia and neurological reaction and thus leading to lesion. Thus, instead of taking skeletal or neurophysiological approach. it seeks to fundamentally resolve the cause of the flow of the life force and blood in muscles not being activated. As a result Oriental medicine attributes the main cause of musculoskeletal system diseases to muscle tension and its build-up that stem from an individual's long formed chronicle habit and work environment. This approach considers not only the social structure aspect including companies owners and work environment that the existing methods have looked at, but also individual workers' responsibility and their environmental factors. Hence, this is a step forward method. 3 The diagnosis of musculoskeletal diseases under Oriental medicine is characterized by the fact that an Oriental medicine doctor uses not only photos taken by himself, but also various detection devices to gather information and pass comprehensive judgment on it. Thus, it is the core of diagnosis under Oriental medicine to develop diagnosing devices matching the characteristics of information to be induced and to interpret information so induced from the views of Oriental medicine. Diagnosis using diagnosing devices values the whole state of a patient and formal abnormality alike, and the whole balance and muscular state of a patient serves as the basis of diagnosis. Hence, this method, instead of depending on the information gathered from devices under Western medicine, requires devices that provide information on the whole state of a patient in addition to the local abnormality information that X-ray. CT, etc., can offer. This method sees muscle as the central part of the abnormality in the musculoskeletal system and thus requires diagnosing devices enabling the muscular state. 4. The diagnosing device using moire topography under Oriental medicine has advantages below and can be used for diagnosing musculoskeletal system diseases with industrial workers . First, the device can Provide information on the body in an unbalanced state. and thus identify the imbalance and difference of height in the left and right stature that a patient can not notice at normal times. Second, the device shows the twisting of muscles or induration regions in a contour map. This is not possible with existing shooting machines such as X-ray, CT, etc., thus differentiating itself from existing machines. Third, this device makes it possible for Oriental medicine to take its unique approach to the abnormality in the musculoskeletal system. Oriental medicine sees the state and imbalance state in muscles as major factors in determining the lesion of musculoskeletal system, and the device makes it possible to shoot the state of muscles in detail. In this respect, the device is significant. Fourth, the device has an advantage as non-aggression diagnosing device.

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Taxonomical Classification and Genesis of Jeju Series in Jeju Island (제주도 토양인 제주통의 분류 및 생성)

  • Song, Kwan-Cheol;Hyun, Byung-Geun;Moon, Kyung-Hwan;Jeon, Seung-Jong;Lim, Han-Cheol;Lee, Shin-Chan
    • Korean Journal of Soil Science and Fertilizer
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    • v.43 no.2
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    • pp.230-236
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    • 2010
  • Jeju Island is a volanic island which is located about 96 km south of Korean Peninsula. Volcanic ejecta, and volcaniclastic materials are widespread as soil parent materials throughout the island. Soils on the island have the characteristics of typical volcanic ash soils. This study was conducted to reclassify Jeju series based on the second edition of Soil Taxonomy and to discuss the formation of Jeju series in Jeju Island. Morphological properties of typifying pedon of Jeju series were investigated, and physico-chemical properties were analyzed according to Soil survey laboratory methods manual. The typifying pedon has dark brown (10YR 3/3) silt clay loam A horizon (0~22 cm), strong brown (7.5YR 4/6) silty clay BAt horizon (22~43 cm), brown (7.5YR 4/4) silty clay Bt1 horizon (43~80 cm), brown (7.5YR 4/6) silty clay loamBt2 horizon (80~105 cm), and brown (10YR 5/4) silty clay loam Bt3 horizon (105~150 cm). It is developed in elevated lava plain, and are derived from basalt, and pyroclastic materials. The typifying pedon contains 1.3~2.1% oxalate extractable (Al + 1/2 Fe), less than 85%phosphate retention, and higher bulk density than 0.90 Mg $m^{-3}$. That can not be classified as Andisol. But it has an argillic horizon from a depth of 22 to 150 cm, and a base saturation (sum of cations) of less than 35% at 125 cm below the upper boundary of the argillic horizon. That can be classified as Ultisol, not as Andisol. Its has 0.9% or more organic carbon in the upper 15 cm of the argillic horizon, and can be classified as Humult. It dose not have fragipan, kandic horizon, sombric horizon, plinthite, etc. in the given depths, and key out as Haplohumult. A hoizon (0~22 cm) has a fine-earth fraction with both a bulk density of 1.0 Mg $cm^{-3}$ or less, and Al plus 1/2 Fe percentages (by ammonium oxalate) totaling more than 1.0. Thus, it keys out as Andic Haplohumult. It has 35% or more clay at the particle-size control section, and has thermic soil temperature regime. Jeju series can be classified as fine, mixed, themic family of Andic Haplohumults, not as ashy, thermic family of Typic Hapludands. In the western, and northern coastal areas which have a relatively dry climate in Jeju Island, non Andisols are widely distributed. Mean annual precipitation increase 110 mm, and mean annual temperature decrease $0.8^{\circ}C$ with increasing elevation of 100m. In the western, and northern mid-mountaineous areas Andisols, and non Andisols are distributed simultaneously. Jeju series distributed mainly in the western and northern mid-mountaineous areas are developed as Ultisols with Andic subgroup.

Effects of Activation Treatments and Culture Condition on In Vitro Development of Caprine In Vivo and In Vitro Oocytes (재래산양의 체내 및 체외유래 난자의 활성화 처리방법 및 배양조건이 단위발생란의 체외발달에 미치는 영향)

  • Park H. S.;Kim T. S.;Lee Y. H.;Jung S. Y.;Lee M. Y.;Jin J. I.;Park J. K.;Lee J. S.;Kim C. H.
    • Reproductive and Developmental Biology
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    • v.28 no.3
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    • pp.181-185
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    • 2004
  • This study was conducted to examine whether activation treatments, source of oocytes and culture conditions affect in vitro developmental ability of caprine oocytes. Mature Korean native goats were pretreated with intravaginal CIDR for 10 days. The goats were then treated with a single intramuscular injection of 1,000 IU PMSG on Day 8 or twice daily injection of a total of 70 mg FSH for 3 days from Day 8 of CIDR insertion for superovulation. All the goats were injected with 10 mg PGF/sub 2a/ on Day 8 and 400 IU hCG on Day 10 of CIDR. Oocytes were surgically collected by oviduct flushing(in vivo maturation) or direct follicle aspiration(in vitro maturation) through mid-ventral incision at 35 h after hCG injection. Fifteen to twenty oocytes were placed in TCM-199 medium containing 25 mM Hepes and hormones under mineral oil at 39℃ in a humudified atmosphere of 5% CO₂ in air for 22 to 24 h. After maturation, the oocytes were activated by electric stimulation or ionomycin + 6-DMAP. The activated oocytes were then cultured in M16, TCM-199 and mSOF media supplemented with proteins at 39℃ for 6 to 7 days. Activation treatments did not affect cleavage of the oocytes. The cleavage rates were 64.1% (41/64) in oocytes activated by electric stimulation and 76.5% (218/285) in oocytes activated by ionomycin + 6-DMAP. The proportion of development to blastocyst was 15.6% (34/218) in oocytes activated by ionomycin + 6-DMAP, but activation by electric stimulation did not support embryos developed beyond morula stage. There were no differences in the cleavage rates of activated oocytes experiencing in vivo (86.8%, 66/76) and in vitro maturation (69.0%, 127/184). However, the development rate to blastocyst stage was significantly (P<0.05) higher for oocytes matured in vivo (50.0%, 33/66) compared to in vitro (0.8%, 1/127). Culture conditions did not affect the cleavage of -activated oocytes. The cleavage rates were 51.6% (49/95) in M16, 64.3% (18/28) in TCM-199 and 81.0% (145/179) in mSOF, respectively. By contrast, the development rate of activated oocytes to stage was greater (P<0.05) for oocytes cultured in mSOF medium (23.4%, 34/145) than in M16 or TCM-199 (0.0%). Our results suggest that source of oocytes and culture conditions are major factors affecting in vitro development of caprine parthenogenetic oocytes.

The Effects of Isopropyl 2-(1,3-dithioetane-2-ylidene)-2-[N-(4-methyl-thiazol-2-yl)carbamoyl]acetate (YH439) on Potentiated Carbon Tetrachloride Hepatotoxicity (상승적 화학적 간독성에 미치는 YH439의 영향)

  • Kim, Sang-Geon;Cho, Joo-Youn
    • The Korean Journal of Pharmacology
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    • v.32 no.3
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    • pp.407-416
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    • 1996
  • The reactive intermediates formed during the metabolism of therapeutic agents, toxicants and carcinogens by cytochromes P450 are frequently capable of covalently binding to tissue macromolecules and causing tissue damage. It has been shown that YH439, a congener of malotilate, is effective in suppressing hepatic P450 2E1 expression. The present study was designed to further establish the mechanistic basis of YH439 protection against toxicant by assessing its effects against chemical-mediated potentiated hepatotoxicity. Retinoyl palmitate (Vit-A) pretreatment of rats for 7 days substantially enhanced carbon tetrachloride hepatotoxicity, as supported by an ${\sim}5-fold$ increase in serum alanine aminotransferase (ALT) activity, as compared to $CCl_4$ treatment alone. The elevation of ALT activity due to Vit-A was completely blocked by the treatment of $GdCl_3$ a selective inhibitor of Kupffer cell activity. Concomitant pretreatment of rats with both YH439 and Vit-A resulted in a 94% decrease in Vit-A-potentiated $CCl_4$ hepatotoxicity. YH439 was also effective against propyl sulfide-potentiated $CCl_4-induced$ hepatotoxicity. Whereas propyl sulfide (50 mg/kg, 7d) enhanced $CCl_4-induced$ hepatotoxicity by >5-fold, relative to $CCl_4$ treatment alone, concomitant treatment of animals with both propyl sulfide and YH439 at the doses of 100 and 200 mg/kg prevented propyl sulfide-potentiated $CCl_4$ hepatotoxicity by 35% and 90%, respectively. Allyl sulfide, a suppressant of hepatic P450 2E1 expression, completely blocked the propyl sulfide-enhanced hepatotoxicity, indicating that propyl sulfide potentiation of $CCl_4$ hepatotoxicity was highly associated with the expression of P450 2E1 and that YH439 blocked the propyl sulfide-enhanced hepatotoxicity through modulation of P450 2E1 levels. Propyl sulfide- and $CCl_4-induced$ stimulation of lipid peroxidation was also suppressed by YH439 in a dose-related manner, as supported by decreases in malonedialdehyde production. The role of P450 2E1 induction in the potentiation of $CCl_4$ toxicity and the effects of YH439 were further evaluated using pyridine as a P450 2E1 inducer. Pyridine pretreatment substantially enhanced the $CCl_4$ hepatotoicity by 23-fold, relative to $CCl_4$ alone. YH439, however, failed to reduce the pyridine-potentiated toxicity, suggesting that the other form(s) of cytochroms P450 inducible by pyridine, but not suppressible by YH439 treatment, may play a role in potentiating $CCl_4-induced$ hepatotoxicity. YH439 was capable of blocking cadmium chloride-induced liver toxicity in mice. These results demonstrated that YH439 efficiently blocks Vit-A-enhanced hepatotoxiciy through Kupffer cell inactivation and that the suppression of P450 2E1 expression by YH439 is highly associated with blocking of propyl sulfide-mediated hepatotoxicity.

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