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Effects of Sperm Pretreatments and In vitro Culture Systems on Development of In vitro Fertilized Embryos Derived from Prepubertal Boer Goat Oocytes in China

  • Lv, Lihua;Yue, Wenbin;Liu, Wenzhong;Ren, Youshe;Li, Fuzhong;Lee, Kyung-Bon;Smith, George W.
    • Asian-Australasian Journal of Animal Sciences
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    • v.22 no.7
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    • pp.969-976
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    • 2009
  • Use of oocytes from prepubertal animals for in vitro embryo production holds potential application for reducing generation intervals and increasing genetic progress through embryo transfer. The objective of these studies was to compare the effect of three sperm pretreatments (prior to in vitro fertilization) and seven embryo culture protocols on fertilization rate and (or) subsequent development of in vitro fertilized embryos derived from oocytes harvested from ovaries of 1-6 month old prepubertal Boer goats in China. Cleavage rates were highest for embryos fertilized with heparin-treated versus calcium ionophore- or caffeine-treated sperm. Similar rates of blastocyst development were observed using heparin- and ionophore-treated sperm, which were higher than obtained with caffeine-treated sperm. No differences in cleavage or blastocyst rates were observed following embryo culture in basal medias (synthetic oviductal fluid (SOF), Charles Rosenkrans 1 (CR1) or tissue culture medium-199 (TCM-199)) containing 10% fetal bovine serum (FBS). Cumulus or oviductal cell co-culture did not enhance cleavage or blastocyst rates relative to culture in SOF+10% FBS. Replacement of FBS in SOF medium with 0.3% BSA increased cleavage rates, but did not increase rates of blastocyst development. Sequential culture in SOF+0.3% BSA followed by SOF+10% FBS increased blastocyst yield versus continuous culture in SOF+10% FBS and tended to increase blastocyst yield versus continuous culture in SOF+0.3% BSA. These results demonstrate a pronounced effects of sperm pretreatments and in vitro embryo culture systems on rates of blastocyst development and provide a potential protocol (sperm pretreatment with heparin and sequential embryo culture in SOF+0.3% BSA followed by SOF+10% FBS) for generation of the significant numbers of in vitro produced blastocysts from oocytes of prepubertal Boer goats necessary for application of embryo transfer in rural regions of China for distribution of Boer goat genetics.

The Study on Wound Healing in Rabbit Skins by Low-intensity Laser Irradiation (저강도 레이저 조사에 의한 가토 피부의 상처 치유에 관한 연구)

  • 김식현;전진석
    • Biomedical Science Letters
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    • v.6 no.2
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    • pp.119-129
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    • 2000
  • The skin is an organ that has many important roles, including protection against infection, regulation of temperature and fluid loss, and sensory function. Injury to the skin, wound repair normally involves: (1) balanced activity of inflammation, (2) the re-epithelial phase and (3) the matrix formation of remodeling phase. Thus, skin wound healing is a finely controlled biological process involving a series of complex cellular interactions. Laser therapy is being implemented with increasing frequency in medicine. Low intensity laser is one that is capable of producing an energy density so low that any biologic alterations are the result of direct irradiation effect, not thermal events. This study was designed to evaluate the efficacy of low intensity laser therapy on skin wound healing in rabbits. A total of 10 male rabbits (New Zealand White Rabbit), age 8 weeks were used. Skin wound were surgically created dorso-lateral on the flank of 10 rabbits (2$\times$2 cm/damage areas). The experimental animals were treated with 5Hz (830 nm wave length) low-intensity laser (MILTA-01 Model) daily for 10 min (1.6 J/$cm^2$) for 12 days. Control animals were sham treated with the laser head. Laser irradiation animals showed a complete remodeling of the epithelial layer, a positive repair of connective tissues, and enhanced the wound closure rate over time as compared to the control animals. Especially, laser irradiation groups improved fibroblast activity, cellular content, granulation tissue formation, and collagen deposition which is resulted in improving the tensile strength of the wound. These findings suggest that laser photostimulation could accelerate healing of open wound in rabbits, and may be benefit in the treatment of open wound, including decubitis ulcers.

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THE EFFECTS OF SURFACE CONTAMINATION ON THE SHEAR BOND STRENGTH OF COMPOMER

  • Heo, Jeong-Moo;Lee, Su-Jong;Im, Mi-Kyung
    • Proceedings of the KACD Conference
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    • 2001.11a
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    • pp.577-577
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    • 2001
  • The lastest concepts in bonding are "total etch", in which both enamel and dentin are etched with an acid to remove the smear layers, and "wet dentin" in which the dentin is not blown dry but left moist before application of the bonding primer. Ideally, the application of a bonding agent to tooth structure should be insensitive to minor contamination from oral fluids. Clinically contaminations such as saliva, gingival fluid, blood and handpiece lubricant are often encountered by dentists during preparation of a restoration. The aim of this study was to evaluate the effect of contamination by hem-ostatic agents on shear bond strength of compomer restorations. One hundred and ten extracted human maxillary and mandibular molar teeth were collected. The teeth were cleaned from soft tissue remnant and debris and stored in physiologic solution until they were used. Small flat area on dentin of the buccal surface were wet ground serially with 400, 800 and 1200 abrasive paper on automatic polishing machine. The teeth were randomly divided into 11 groups. Each group was conditioned as follows: Group 1 : Dentin surface was not etched and not contaminated by hemostatic agents. Group2 : Dentin surface was not etched but was contaminated by Astringedent (Ultradent product Inc., Utah, U.S.A.). Group3 : Dentin surface was not etched but was contaminated by Bosmin (Jeil Phann, Korea.). Group4 : Dentin surface was not etched but was contaminated by Epri-dent (Epr Industries, NJ, U.S.A.). Group5: Dentin surface was etched and not contaminated by hemostatic agents. Group 6 : Dentin surface was etched and contaminated by Astringedent. Group7 : Dentin surface was etched and contaminated by Bosmin. Group8 : Dentin surface was etched and contaminated by Epri-dent. Group9 : Dentin surface was contaminated by Astringedent. The contaminated surface was rinsed by water and dried by compressed air. Group10 : Dentin surface was contaminated by Bosmin. The contaminated surface was rinsed by water aud dried by compresfed air. Group 11 : Dentin surface was contaminated by Epri-dent. The contaminated surface was rinsed by water and dried by compresfed air. After surface conditioning, F2000 was applicated on the conditoned dentin surface. The teeth were thermocycled in distilled water at $5^{\circ}C\;and\;55^{\circ}C$ for 1000 cycles. The samples were placed on the binder with the bonded compomer-dentin interface parallel to the lmife-edge shearing rod of the Universal testing machine(Zwick 020, Germany) running at a cross head speed of 1.0mmimin. There were no significant differences in shear bond strength between groups 1 and group 3 and 4, but group 2 showed significant decrease in shear bond strength compared with group 1. There were no significant differences in shear bond strength between group 5 and group 7 and 8, but group 6 showed significant decrease in shear bond strength compared with group 5. There were no significant differences in shear bond strength between group 5 and group 9, 10 and 11.

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The Frequency of Detecting Prevotella intermedia and Prevotella nigrescens in Korean Adult Periodontitis Patients (한국인 치주 감염 환자에서의 Prevotella intermedia와 Prevotella nigrescens의 발현빈도)

  • Peck, Seung-Yup;Ku, Young;Rhyu, In-Cheol;Hahm, Byung-Do;Han, Soo-Boo;Choi, Sang-Mook;Chung, Chong-Pyoung
    • Journal of Periodontal and Implant Science
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    • v.30 no.2
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    • pp.419-429
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    • 2000
  • Prevotella intermedia has been implicated as a potent pathogen in many kinds of periodontal, pulpal and periapical diseases. However, it has been isolated from periodontally healthy adults and from edentulous children as well. The intraspecies heterogeneity of Prevotella intermedia has been demonstrated in early studies and finally Shah & Gharbia confirmed the existence of 2 DNA homology groups and proposed dividing Prevotella intermedia into 2 species, Prevotella intermedia and Prevotella nigrescens. This study was designed to examine the frequency of Prevotella intermedia and Prevotella nigrescens in diseased periodontal pockets and healthy gingival sulcus of Korean people by PCR based on 16s ribosomal DNA sequence. One hundred adults who had adult periodontitis but not taken any periodontal treatment or antibiotics during previous 6 months and 50 adults who had healthy periodontal tissue were selected for this study. The sulcular fluid was collected into VMGA by sterilized paper point and diluted to 1,000 times in anaerobic chamber. $100{\mu}{\ell}$ of sample was cultured in $37^{\circ}C$ for 10 days. Among the bacterial colonies, BPB were selected and cultured in BHI broth and then Prevotella intermedia was identified through Gram staining and biochemical test. Identified Prevotella intermedia was cultured again and centrifuged. DNA was extracted from the pellet using several reagents. PCR was performed by previously designed primer. The results were followed. 1. BPB were isolated from 39 of 100 samples of diseased periodontal pockets(39%). 2. Prevotella intermedia was identified from 24 of 39 BPB samples. 3. Among 24 Prevotella intermedia, 21 were confirmed as Prevotella inter - media(87.5) and 2 were confirmed as Prevotella nigrescens(8.33%). 4. BPB were isolated from 9 of 50 samples of periodontally healthy patients. Among them only two were identified as Prevotella intermedia, that is, one was confirmed as Prevotella intermedia and the other was Prevotella nigrescens.

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Effects of Cyclobuxine D on Carrageenin-induced Pleurisy and Croton Oil-induced Granuloma Pouch in Rats (흰쥐의 Carrageenin 유발 늑막염과 Croton oil 유발 육아종양에 미치는 Cyclobuxine D의 영향)

  • Lee, Jong-Hwoa;Park, Young-Hyun;Cho, Byung-Heon;Kim, Yu-Jae;Kim, Jong-Bae;Kim, Chung-Mok;Kim, Chun-Sook;Cha, Young-Deog;Kim, Young-Suk
    • The Korean Journal of Pharmacology
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    • v.23 no.2
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    • pp.151-157
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    • 1987
  • Cyclobuxine D, a steroidal alkaloid, was extracted from Buxus microphylla var. koreana Nakai. The effects of cyclobuxine D on carrageenin-induced pleurisy and croton oil-induced granuloma pouch in rats was investigated and compared with those of aspirin, hydrocortisone ana dexamethasone. Intrapleural injection of 2% carrageenin caused the accumulation of exudate. The rate of plasma exudation, measured by the exuded dye amounts for 20 min in the pleural cavity after intravenous injection of pontamine sky blue, showed a peak at 5 hr. Cyclobuxine D (5, 20 and 50 mg/kg, i.p.) suppressed dose-dependently the accumulation of the pleural exudate and the exudation of dye. Among several methods used for screening and evaluation anti-inflammatory agents, granuloma pouch technic introduced by Hans Selye (Hans seyle, 1953) is considered as a simple and reliable method. An air pocket was produced in the subcutaneous tissue of the interscapular region by injection of 1 ml of 1% croton oil as irritant. Inflammatory exudate accumulated in the pouch during the succeding 14 days. Cyclobuxine D (5 and 20 mg/kg) decreased fluid volume in pouch and weight of pouch wall in granulomatous inflammation.

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Study on Radioprotection of Alliin and Damage Mechanism in Hepatocyte After Irradiation (방사선 피폭 시 간세포의 손상 기전과 알리인의 방어효과)

  • Ji, Tae-Jeong;Kim, Won-Tae
    • Journal of radiological science and technology
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    • v.39 no.4
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    • pp.623-630
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    • 2016
  • Liver tissue damage by a radiation exposure caused a jaundice and ascitic fluid e form harden atrophy. The reason for this lies in morphological damage of a liver cells. This study tried that observe damage mechanism of the cell organelles. It was especially observed mitochondria, endoplasmic reticulum and nuclear membrane associated with energy metabolizable. also, This study had with a radio-protector development research at the same time. Radio-protector was used to alliin that has an blood flow increase. Cell observation make used of transmission electron microscope(TEM). The result of an experiment, 7Gy of whole body irradiation was caused an inflammation in cell organelles and hypertrophy of the nucleus membrane. After 20 days, The hepatocyte has been observed in a damaged membrane on peroxisome, mitochondria and vacuole of the cell organelles. After 30 days, The hepatocyte has been observed in disconnected ribosomes on a rough endoplasmic reticulum. There was looked a giant lipoblast. There was clearly normal observed a mitochondria and nucleus membrane in the hepatocyte after alliin injection. aslo, It was no damaged the nucleus membrane. therefore, It was identified portion a radio-protector effect from alliin.

The Flow Behavior of Skin Collagen (피부조직 콜라겐의 유동 특성)

  • Kim, Young-Ho;Park, Eun-Ji;Yang, Ryung
    • Korean Journal of Food Science and Technology
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    • v.27 no.4
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    • pp.576-581
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    • 1995
  • To obtain the basic information for the effective use of collagen, the flow behavior of collagen extracted from skin tissue was studied. The viscosity of collagen varied with sex, age and the kinds of collagen by extraction method. Regardless of the kinds of collagen, the viscosity of collagen extracted from $6{\sim}12$ week old rat was relatively high. In case of the same age, the viscosity showed higher in female than in male rat and in acid soluble collagen than in insoluble collagen. The solution of the collagen showed the characteristics of Bingham plastic and thixotropic fluid, and the viscosity varied distinctly with temperature, pH, ethanol concentration and collagen concentration. As collagen concentration increased to 6%, the consistency of acid soluble- and insoluble collagen showed a tendency to increase linearly(r = 0.972 for acid soluble collagen, r = 0.957 for insoluble collagen). In that range of collagen concentration, the increasing velocity of consistency was higher in acid soluble collagen than in insoluble collagen. The consistency of collagen solution was decreased according to temperature rising. In case of acid soluble collagen, the consistency is decreased abruptly between $30{\sim}40^{\circ}C$. According to pH variation, the consistency of acid soluble collagen showed biphasic phenomenon, though the consistency of insoluble collagen was found not to be influenced by pH. The consistency of acid soluble- and insoluble collagen according to ethanol concentration showed high between $40{\sim}60%$ of ethanol concentration.

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Subsequent Embryo Transfers (SET) on Day 2 and Day 5: It's Safety and Effectiveness (난자채취 2일과 5일에 연속으로 실시한 배아이식의 안전성과 효과)

  • Park, Kee-Sang;Song, Hai-Bum;Lee, Taek-Hoo;Jeon, Sang-Sik
    • Clinical and Experimental Reproductive Medicine
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    • v.27 no.2
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    • pp.165-172
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    • 2000
  • Objective: In vitro fertilization (IVF) and a prolonging the time of culture may be helpful in establishing a viable pregnancy through a selection effect. Some embryos do not develop beyond the 4-cell stage and some may not develop to the blastocyst stage. We have evaluated the safety of SET and the outcomes of pregnancy. Methods: Sperms were treated with Ham's F-10 supplemented with 10% human follicular fluid (hFF). oocytes or fertilized oocytes were cultured in Dulbecco's Modified Eagle Medium (DMEM) with 10% or 20% hFF respectively. Up to five oocytes were inseminated with approximately 200,000 sperm cells/2 ml in each well. Fertilization was examined in the following morning and fertilized oocytes were co-cultured until embryo transfer. Vero cells for co-culture were prepared in Tissue Culture Medium - 199 (TCM-199) with 10% fetal bovine serum. At the two to four cell and blastocyst on day 2 and day 5, embryo and blstocyst grading were evaluated. Pregnancy rate was determined after transfer of human embryos at the two to four cell stage on day 2 (Group I) or subsequent transfer of embryos on day 2 and at the blastocyst stage on day 5 (Group II). For statistical analysis, Student's t-test and Chi-square (${\chi}^2$_test) were used. Results were considered statistically significant when p value was less than 0.05. Results: No differences was found in the fertilization between Group I (81.0%, 98/121) and Group II (81.8%, 180/220). In case of cleavage rate, no difference was found in Group I (95.9%, 94/98) and Group II (97.8%, 174/178). However, the rate of-clinical pregnancy was significantly higher (p=0.014) in Group II (66.7%, 12/18) than in Group I (26.3%, 5/19). Conclusion: The results of this study showed that SET is safe and effective, and significantly increases the pregnancy rate.

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Diffuse Pulmonary Alveolar Hemorrhage and Interstitial Pneumonitis after Subcutaneous Injection of Atelocollagen for Cosmetic Purpose : Two Case Reports (미용 목적으로 Atelocollagen 주입 후에 발생한 미만성 폐포출혈과 간질성 폐렴 2례)

  • Ko, Young Chun;Lim, Sung Chul;Park, Kyung Hwa;Kim, Jeong Soon;Kim, Kyu Sik;Kim, Yu Il;Kim, Young Chul;Yoon, Sung Ho;Lee, Seung Il;Park, Kyung Ok
    • Tuberculosis and Respiratory Diseases
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    • v.56 no.3
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    • pp.308-314
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    • 2004
  • Atelocollagen have been used recently in skin and other soft tissue defect regions more than silicone fluid because of the low incidence of an immune reaction and complications. Several cases of acute pneumonitis after a subcutaneous injection of silicone have been reported. The symptoms were dyspnea, fever, chest pain and hemoptysis. Previous reports have explained the pathophysiology of acute pneumonitis to a pulmonary microembolism and cellular inflammation. We experienced two cases of an acute interstitial pneumonitis and pulmonary hemorrhage after a subcutaneous injection of atelocollagen. They were all healthy young women and complained of dyspnea, fever and blood tinged sputum. The chest radiography and computerized tomography showed a bilateral ground glass opacity in both lung fields. One case recovered completely with conservative treatment but the clinical course of the other case was aggravated to the degree that invasive positive pressure ventilation therapy was required. We report a rare case of a diffuse pulmonary alveolar hemorrhage and an interstitial pneumonitis after the subcutaneous injection of atelocollagen for cosmetic purposes.

Aspirin Reduces Acute Lung Injury in Rats Subjected to Severe Hemorrhage (Aspirin이 출혈성 쇼크로 인한 급성 폐손상에 미치는 효과)

  • Shin, Tae Rim;Lee, Dong Uk;Park, Yoon-Yub
    • Tuberculosis and Respiratory Diseases
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    • v.54 no.5
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    • pp.522-531
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    • 2003
  • Background : Hemorrhagic shock and trauma are two of the most common causes of acute lung injury. The activation of cyclooxygenase is one of the important causes of acute lung injury. This study investigated the effect of aspirin, a well-known cyclooxygenase inhibitor, on severe hemorrhage-induced acute lung injury in rats. Methods : The hemorrhagic shock was induced by withdrawing blood; 20ml/kg of B.W., through the femoral artery in 5 min. The mean arterial pressure was recorded through the femoral artery on a polygraph. Results : In the present investigation, the lung tissue myeloperoxidase activity, protein contents and leukocyte counts, in bronchoalveolar lavage fluid, increased significantly 2 and 24 h after the hemorrhage induction. Although the decreased mean arterial pressure spontaneously recovered, acute lung injury occurred after severe hemorrhage. These changes were effectively prevented by a single intravenous injection of aspirin (10 mg/kg of B.W.) 30 min before the hemorrhage. Conclusion : These results suggest that severe hemorrhage-induced acute lung injury is mediated, in part, by the activation of cyclooxygenase. Furthermore, pretreatment of aspirin in acute lung injury-prone patients, or prophylactic treatment of aspirin to the patients with precipitating conditions, could be helpful in the prevention of acute lung injury.