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Studies on the Liquid and Freezing storage of Boar Semen III. Effects of Dilutor on Livability and Fertility of Liquid Semen and Effects of Dilution and Vessel on Livability of Frozen Semen (돈정액의 액상 및 동결보존에 관한 연구. III. 보존액이 액상정액의 정자생존성과 수태율에 미치는 영향과 희석방법과 용기가 동결정액의 생존성에 미치는 영향)

  • 임경순;정장용
    • Korean Journal of Animal Reproduction
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    • v.3 no.1
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    • pp.30-35
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    • 1979
  • A, B and C dilutors were used to make Ka (A plus B (1 : 1)) and Na (B plus C(1 : 1)) dilutors in this experiment. Three aliqots of semen were respectivly diluted 1 : 1 and 1 : 2 (semen: dilutor) with Ka, Na and C dilutors and stored at 5$^{\circ}C$ for 7 days in order to study their livability during storage. Fertility was checked for the diluted semen with Ka, Na and C dilutors. Whole semen and extended semen with Na dilutos with and without DMSO were cold shocked at various temperatures for 10 min. Effects of different 1st and 2nd dilution with A, B, C and Na dilutors and of vessels on freezability of spermatozoa were investigtigated. 1. Extended semen 1 : 2 with Na and C dilutors showed highest live sperm index during storage for 7 days at 5$^{\circ}C$. 2. The components of Na dilutor per 100$m\ell$ were skim milk 2.5g, trisaminomethane 0.54g, citric acid 0.265g, glucose 2.835g, fructose 1.5g, sodium lauryl sulfate, 0.08g, penicillin 0.06g, streptomycin 0.075g, and egg yolk 10$m\ell$. 3. Fertility of diluted semen was higher than that of whole semen. Ka dilutor showed higher fertility than Na and C dilutors, and there was no difference in the fertility between Na and C dilutors. 4. Na dilutor with DMSO showed slightly higher livability than Na dilutor without DMSO during storage for 7 days at 5$^{\circ}C$. 5. Cold shock at 1$0^{\circ}C$ for 10 min. decreased greatly the sperm livalility of whole semen but not of extended semen with Na dilutor. Addition of DMSO to Na dilutor has no effect in prevention of cold shock. 6. The extended semen with C. C dilutor (1st and 2nd dilution with C and C dilutor) showed higher post-thawing sperm livability than A.A and Na. B dilutors. Na. B dilution shwed higher post-thawing sperm livability than A.A dilution. There was no difference in the post-thawing livability between semen in 1$m\ell$ straw and 10$m\ell$ aluminium package.

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Expression of EGFP in Bovine Embryos after Intracytoplasmic Sperm Injection using Spermatozoa Co-cultured with Exogenous DNA (소 난자에 있어서 외래유전자가 도입된 정자의 직접 주입에 의한 EGFP 의 발현)

  • Lee, . H. C.;S. J. Uhm;S. Y. Ann;H. J. Chung;Park, H. D.;Lee, H. T.;K. S. Chung
    • Korean Journal of Animal Reproduction
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    • v.24 no.4
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    • pp.439-449
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    • 2000
  • This study was to investigate the expression of transgene after co-injection of spermatozoon and EGFP gene into mature oocytes in cattle. From frozen semen, spermatozoa were treated by DTT with 0.03% Tween-20, freezing and thawing or 0.02% Triton X-100 to disrupt their plasma membranes. The sperm injected oocytes were co-cultured with bovine oviduct epithelial cells in CRlaa, and expression of EGFP in embryos were observbed under epifluorescent microscope. Two pronuclei were formed in oocytes injected with sperm treated by DTT (44.6%), DTT-Tween-20 (48.4%), DTT-freezing (44.4%) and DTT-Triton X-100 (42.9%). Cleavage and blastocyst formation rates of bovine oocytes which injected with sperm treated by DTT, DTT-Tween-20, DTT-freezing, and DTT-Triton X-100 were 49.1, 58.5, 43.9, and 48.4% and 10.2, 13.0, 6.8, and 6.5%, respectively. Although the most of embryos were showing mosaicism, embryos expressing EGFP gene were observed in all treated groups. Therefore, these results indicate that membrane disrupted sperm could interact with exogenous DNA, and that this procedure may be useful to introduce foreign gene into bovine oocytes.

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Study on Development of Canine Oocytes Treated by In Vitro Fertilization and ICSI

  • Park, Ji-Hoon;Chung, Young-Ho;Kim, Sang-Keun
    • Reproductive and Developmental Biology
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    • v.35 no.2
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    • pp.137-141
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    • 2011
  • These study was to investigate the in vitro fertilization and viability of fresh and vitrified oocytes. Also, the developmental capacity of IVF and intracytoplasmic sperm injection (ICSI) oocytes were investigated. Then vitrification was performed with the use of 20% ethylene glycol + 20% DMSO + 0.5 M sucrose + 10% FCS + TCM-199 medium. Vitrification immature oocytes are cultured in vitrification solution for 10 min afterwards transferred to expose at room temperature for 5 min. and transferred to the ice water for 5 min. The oocytes were sealed in a 1.0 mm straw and placed in a $LN_2$ container. Frozen oocytes were rapidly thawed in a water bath at $30{\sim}35^{\circ}C$, and then placed in TCM-199 medium containing 0.5 M sucrose for 5 min each, respectively, at $38^{\circ}C$. After being washed for 2~3 times, using fresh medium the oocytes were cultured in TCM-l99 medium supplemented with 5% FCS at $38^{\circ}C$ in 5% $CO_2$ and air. The normal morphology of fresh and vitrified-thawed oocytes were $87.1{\pm}2.1%$ and $54.8{\pm}2.5%$, respectively. The viability rates of fresh and vitrified-thawed oocytes were $70.0{\pm}2.2%$ and $41.9{\pm}2.6%$, respectively. Viability rates of vitrified-thawed oocytes were lower than that of fresh follicular oocytes (p<0.05). The in vitro maturation rates of fresh and vitrified oocytes were $45.1{\pm}3.6%$ and $28.9{\pm}4.4%$, respectively. The IVF rates of fresh follicular and vitrified-thawed oocytes were 34.00.2% and $20.2{\pm}2.6%$, respectively. The in vitro maturation and fertilization rates of vitrified-thawed oocytes were lower than those of the fresh follicular oocytes (p<0.05). A total of 350 oocytes were fixed and stained after co-incubation with spermatozoa, of which 88 had identifiable nuclear material. After IVF for 20 hrs, $25.1{\pm}3.4%$ of the oocytes found to have been penetrated by spermatozoas. Oocytes were fixed and stained after ICSI, and 105 oocytes contained identifiable nuclear material. After IVF and ICSI for 20 hrs, $34.3{\pm}3.4%$ and $59.0{\pm}2.0%$ of the oocytes were found to have been penetrated by spermatozoas. The developmental rates upon ICSI were significantly higher than those of the IVF method (p<0.05).

Giant Neurofibroma on Both Buttocks (양측 엉덩이의 거대 신경섬유종)

  • Kim, Ji Hoon;Burm, Jin Sik;Kim, Yang Woo;Kang, So Ra;Kim, Hyoung Kyoung
    • Archives of Plastic Surgery
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    • v.36 no.4
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    • pp.512-515
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    • 2009
  • Purpose: Neurofibromatosis(NF) is an autosomal - dominant systemic disease. Up to fifty percent of patients with NF are reported to have concomitant vascular abnormalities. In the resection of a larger NF, the risk of uncontrolled hemorrhage is much higher due to the difficulty of hemostasis of large vessels within the tumor. We ligated the base of the giant NF with a simple loop - shaped ligation before removal of the giant NF in both buttocks. And then we could successfully reduce the amount of hemorrhage during the operation. Methods: A 46 - year - old female patient presented for giant masses of both gluteal area, which has been growing slowly for the last ten years. Each mass was about $30{\times}20cm$ in size. After designing the elliptical resection margin, we tightened the tumor base by using continuous loop - shaped suture ligation(weaving the thread up and down in a loop - shaped pattern, leaving a space of 2 cm between each loop) with a straight needle and prolene 2 - 0. After skin incision, we proceeded the dissection toward the central and inferior side of the mass obliquely while we avoided breaking large vascular sinuses. We resected the tumor in a wedged - shape. Subcutaneous tissue was sutured layer by layer and skin was closed by vertical mattress and interrupted suture. The loop - shaped ligation of the base was removed and compressive dressing was done with gauzes and elastic bandages. Results: Postoperative complications such as infection, hemorrhage, hematoma, and dehiscense did not occur. Perioperatively the patient was sufficiently transfused with five units of blood and two units of fresh frozen plasma. During the subsequent 1 year follow - up, the functional and cosmetic result was excellent. Conclusion: A continuous loop - shaped suture ligation procedure along the base of the giant NF effectively reduced the amount of hemorrhage during the operation, made dissection and ligation of vessels easily and quickly, and shorten the operating time and postoperative recovery time.

Prevalence of Clostridium difficile Infections in Pigs in Jeju (제주도 돼지에서 Clostridium difficile 감염 양상)

  • Kim, Hyeong-Seok;Jung, Ji-Youl;Kang, Sang-Chul;Shin, Bo-Moon;Lee, Eun-Joo;Hwang, Eui-Kyung;Park, Bong-Kyun;Kim, Dae-Yong;Kim, Jae-Hoon
    • Journal of Veterinary Clinics
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    • v.28 no.2
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    • pp.232-235
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    • 2011
  • Clostridium (C) difficile has been recognized as an important emerging pathogen in both humans and animals. The prevalence of C. difficile in rectal feces and frozen colons of 132 pigs with diarrhea from the Jeju Island was investigated by polymerase chain reaction (PCR) to detect C. difficile toxin A and B genes. PCR findings revealed toxin A and B in 5 pigs (3.8%), including 2 suckling pigs, 2 weaned pigs and 1 growing pig. The result of PCR was closely matched histopathologic lesions of C. difficile in large intestines of pigs. Histopathologically, the cecum and colons of C. difficile toxin-positive pigs had severe submucosal and mesocolonic edema. Mucosal lesions ranged from random single cell necrosis and exfoliation to segmental, transmural necrosis of the cecum and colon. According to bacteriology, 4 C. difficile-positive pigs (80%) were co-infected with Salmonella typhimurium.

Assessment of Immune Quality and Pathogen Contamination of Colostrums Collected from Colostrum Banks in Korea (초유은행에서 수거한 초유의 병원체 오염과 면역수준의 평가)

  • Kim, Won-Il;Park, Sang-Yul;Kim, Sang-Jin;Cho, Yong-Il;Hur, Tai-Young;Kim, Nam-Soo
    • Journal of Veterinary Clinics
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    • v.30 no.4
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    • pp.223-229
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    • 2013
  • Because colostrum is considered to be the sole source of passively acquired maternal antibodies for calves, newborn calves must consume colostrum to gain disease resistance during their early years of life. Storage of surplus colostrum from dairy cows right after calving and feeding newborn calves in deficiency of colostrum to assure adequate uptake of IgG for protection of the calf has been a common practice in the bovine production. In the current study, 35 colostrums were randomly collected from 3 colostrum banks located in different regions of Korea and monitored for general bacterial contamination and major bovine pathogens. Immunoglobulin concentrations and BVDV-specific antibodies were also determined to evaluate the immune quality of the colostrums. Moderate to severe bacterial contamination (up to 72,000,000 CFU/ml) was observed in most of the colostrums collected from colostrum banks. General immune quality of the colostrums was under the satisfactory level since most of the colostrums contained less than 50 g/L of IgG, which is the minimum concentration for good quality colostrums. Therefore, colostrum for colostrum bank should be collected at the first 2-3 post-partum milkings according to proper harvesting and handling procedures to guarantee the safety and quality of colostrum. In addition, it was recommended that colostrum should be heat-treated before frozen and stored in the bank because pasteurization at $63^{\circ}C$ for 30 min was very effective reducing the risk of disease transmission without causing significant degradation of immunoglobulins.

The Seasonal Variation of Primary Productivity in the Antarctic Coastal Ecosystems (남극 연안생태계에서 일차생산력의 계절변화)

  • Kim, Hae-Cheol;Yang, Sung-Ryull;Pae, Se-Jin;Shim, Jae-Hyung
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.3 no.2
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    • pp.80-89
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    • 1998
  • To understand the temporal variation of phytoplankton community in the Antarctic coastal ecosystem, physicochemical parameters, chlorophyll a, and primary productivity were measured as a component of the 7th KARP (Korea Antarctic Research Program) in 1994. Data were collected every month between February and December except four months (June-September) when the study area was frozen. Chlorophyll a concentrations ranged from negligible to 3.03 ${\mu}g/l$, averaging 0.63 ${\mu}g/l$. The primary productivity ranged 0.53-18.95 mg C/$m^3{\cdot}day$, and the depth-integrated primary productivity ranged 41.28-560.20 mg C/$m^3{\cdot}day$. A positive relationship was observed between the phytoplankton biomass and irradiance ($r^2$=0.29, p < 0.01). The degree of correlation between the primary productivity and irradiance ($r^2$=0.85, p < 0.001) was significantly higher than that between the phytoplankton biomass and irradiance. However, neither temperature nor inorganic nutrients seem to affect the temporal variation of primary productivity.

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Polarization Precession Effects for Shear Elastic Waves in Rotated Solids

  • Sarapuloff, Sergii A.
    • Proceedings of the Korean Society for Noise and Vibration Engineering Conference
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    • 2013.04a
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    • pp.842-848
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    • 2013
  • Developments of Solid-State Gyroscopy during last decades are impressive and were based on thin-walled shell resonators like HRG or CRG made from fused quartz or leuko-sapphire. However, a number of design choices for inertial-grade gyroscopes, which can be used for high-g applications and for mass- or middle-scale production, is still very limited. So, considerations of fundamental physical effects in solids that can be used for development of a miniature, completely solid-state, and lower-cost sensor look urgent. There is a variety of different types of bulk acoustic (elastic) waves (BAW) in anisotropic solids. Shear waves with different variants of their polarization have to be studied especially carefully, because shear sounds in glasses and crystals are sensitive to a turn of the solid as a whole, and, so, they can be used for development of gyroscopic sensors. For an isotropic medium (for a glass or a fine polycrystalline body), classic Lame's theorem (so-called, a general solution of Elasticity Theory or Green-Lame's representation) has been modified for enough general case: an elastic medium rotated about an arbitrary set of axes. Travelling, standing, and mixed shear waves propagating in an infinite isotopic medium (or between a pair of parallel reflecting surfaces) have been considered too. An analogy with classic Foucault's pendulum has been underlined for the effect of a turn of a polarizational plane (i.e., an integration effect for an input angular rate) due to a medium's turn about the axis of the wave propagation. These cases demonstrate a whole-angle regime of gyroscopic operation. Single-crystals are anisotropic media, and, therefore, to reflect influence of the crystal's rotation, classic Christoffel-Green's tensors have been modified. Cases of acoustic axes corresponding to equal velocities for a pair of the pure-transverse (shear) waves have of an evident applied interest. For such a special direction in a crystal, different polarizations of waves are possible, and the gyroscopic effect of "polarizational precession" can be observed like for a glass. Naturally, formation of a wave pattern in a massive elastic body is much more complex due to reflections from its boundaries. Some of these complexities can be eliminated. However, a non-homogeneity has a fundamental nature for any amorphous medium due to its thermodynamically-unstable micro-structure, having fluctuations of the rapidly-frozen liquid. For single-crystalline structures, blockness (walls of dislocations) plays a similar role. Physical nature and kinematic particularities of several typical "drifts" in polarizational BAW gyros (P-BAW) have been considered briefly too. They include irregular precessions ("polarizational beats") due to: non-homogeneity of mass density and elastic moduli, dissymmetry of intrinsic losses, and an angular mismatch between propagation and acoustic axes.

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Effects of established cell lines on bovine embryo development during in vitro culture (계대세포를 이용한 소 수정란의 체외배양 체계에 관한 연구)

  • Joo, Suk-chun;Lee, Byeong-chun;Lee, Won-yu;Choi, Yun-seok;Hwang, Woo-suk
    • Korean Journal of Veterinary Research
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    • v.37 no.3
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    • pp.647-659
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    • 1997
  • To overcome the difficulties of collecting and culture of primary cell from genital tract on embryonic development, the present study was carried out to investigate the critical effect of cell lines, such as BRL and Vero cell and its conditioned medium on the development of early Korean native cattle embryos in vitro. Oocytes collected from slaughterhouse ovaries were matured in TCM199 containing FSH, estradiol-$17{\beta}$ and FBS with granulosa cell monolayer for 24 hours and then fertilized in vitro using frozen-thawed, heparin-treated spermatozoa in TALP for 30 hours. And then early embryos (1-2cell) were cultured in TCM199 containing 10% FBS with BOEC, Granulosa, BRL, Vera cell monolayers and conditioned medium for 2~3 days. Development to morulae and blastocysts were recorded, also examined the number of blastomeres presented a valuable parameter for the evaluation of embryonic development. The early cleavage rates of in vitro fertilized embryos co-cultured, there was no differences between primary cell and cell lines(p<0.05). The rate of development to the later stage, coculture of BRL cell was significantly higher than that of the primary cell(p<0.05). The rates of development to morula and blastocyst were significantly higher in vero cell than BRL, Granulosa, Oviduct epithelial cell conditioned medium. In the result of effect of serum on development of early bovine embryos, the use of media containing serum were significantly higher than the use of not containing one on development of early and later stage of embryos. The result of number of blastomeres in blastocysts, there is no differences between primary cell and cell lines. The blastocysts from coculture were higher than from conditioned medium in blastomere cells. In summary, these experments have proved that the culture system in TCM199 with BRL, Vero cell monolayers is effective on in vitro development of early bovine embryos, In addition, it is effective to development of bovine embryos that containing serum in conditioned medium, or in co-culture rather than in conditioned medium alone. The use of cell lines opponent to primary cells is effective in bovine embryo culture.

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Clinical Analysis of Surgically Treated Thyroid Nodules (외과적 치료를 시행한 갑상선 결절의 임상적 고찰)

  • Chang Yong-Keun;Kim Kweon-Cheon
    • Korean Journal of Head & Neck Oncology
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    • v.15 no.1
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    • pp.70-75
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    • 1999
  • Objectives: This study was designed to evaluate the methods of diagnosis and treatment of thyroid nodules. Materials and Methods : We performed a clinical review of patients with thyroid nodules, who were surgically treated at the Department of Surgery, College of Medicine, Chosun University from January 1996 to December 1998. Results: 1) The patients were divided into two groups; 65 patients(80.2%) with benign nodular disease and 16 patients(19.8%) with malignant disease. 2) The sex distribution showed a preponderance of females with a ratio of 5.2:1 in benign nodular disease and 3:1 in malignant disease. Benign nodules were more prevalent in patients in their 40's as compared to malignant nodules in patients in their 50's and 60's. 3) The most common duration of illness was 3 months, occurring in 45.7% of the total cases. 4) The most prominent symptom and sign was a palpable nodule in the anterior aspect of the neck. 5) The location of the nodule was ; 41 cases in the right lobe, 29 cases in the left lobe, and 11 cases in both lobes. 6) Thyroid scanning of 40 patients revealed cold nodules in 90.6% of benign nodules and in all malignant disease. 7) Fine needle aspiration cytology were performed in 32 cases of the 81 patients. Comparing with postoperative pathological findings, the results were the same in 87.5% of 32 cases. 8) Intra-operative frozen section study was performed in 56 cases of the 81 patients. Comparing with postoperative pathological finding, the results were same in 96.4% of the 56 cases. 9) The histopathological classification revealed that adenomatous goiters were the most common benign disease and papillary carcinomas were the most common malignant disease. 10) The most frequently employed operation for benign nodules and malignant disease was unilateral total lobectomy. When the metastasis was confirmed, lymphadenectomy and radical neck dissection was performed in malignant disease. 11) Important postoperative complications were transient hoarseness, transient hypocalcemia, hypothyroidism, wound bleeding, and hoarseness. Conclusions: The results of this study suggest that palpable nodules in anterior aspect of neck is revealed cold nodule by thyroid scanning and malignancy by fine needle aspiration cytology, which should be removed surgically. There is no difference in complication and survival rate with type of operation and lymphatic dissection. Therefore, procedure of operation is dependent on the site of nodule and involvement of lymph node.

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