• 제목/요약/키워드: Frozen

검색결과 2,678건 처리시간 0.031초

돈정액의 액상 및 동결보존에 관한 연구. III. 보존액이 액상정액의 정자생존성과 수태율에 미치는 영향과 희석방법과 용기가 동결정액의 생존성에 미치는 영향 (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)

  • 임경순;정장용
    • 한국가축번식학회지
    • /
    • 제3권1호
    • /
    • pp.30-35
    • /
    • 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.

  • PDF

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

  • Lee, . H. C.;S. J. Uhm;S. Y. Ann;H. J. Chung;Park, H. D.;Lee, H. T.;K. S. Chung
    • 한국가축번식학회지
    • /
    • 제24권4호
    • /
    • pp.439-449
    • /
    • 2000
  • 본 연구에서는 정자와 외래유전자인 EGFP 유전자를 공배양한 후 정자직접 주입술로 난자를 수정시켜 EGFP 유전자의 발현을 조사하였다. 정자는 외래유전자의 도입이 용이하도록 동결융해, 0.03% Tween-20과 0.02%의 Triton X-100의 처리를 통하여 정자두부의 원형질막을 제거하여 공시하였다. 수정된 난자는 소 난관상피세포가 포함된 CR1aa 배양액에서 공배양을 통하여 체외발달시켰으며, 난자의 발달에 따라 EGFP 유전자의 발현을 형광 현미경 하에서 조사하였다. 원형질막이 제거된 정자로부터 수정란의 정상수정을 확인하기 위하여 18시간째 2PN 2PB를 조사한 결과, 발생율은 각각 DTT 처리구 44.6%, DTT와 Twen-20 처리구 48.4%, DTT와 동결융해 처리구 44.4%, 그리고 DTT와 Triton X-100 처리구 42.9%였다. 수정란의 초기 배 분할율은 DTT 처리구 49.1 %, DTT와 Tween-20 처리구 58.5%, DTT와 동결융해 처리구 43.9% 그리고 DTT와 Triton X-100 처리구 48.4%였으며, 배반포 형성율은 DTT 처리구 10.2%, DTT와 Tween-20 처리구 13.0%, DTT와 동결융해 처리구 6.8% 그리고 DTT와 Triton X-100 처리구 6.5%였다. 이들 발달된 수정란 중 도입된 EGFP 유전자의 발현율은 DTT 처리구 3.8%, DTT와 Tween-20 처리구 11.1%, DTT와 동결융해 처리구 13.8% 그리고 DTT와 Triton X-100 처리구 8.9%로 나타났으며, 대부분의 발현은 모자이크 형태로 관찰되었다. 따라서 본 연구의 결과에 의하면 소에서 원형질막을 제거한 정자와 외래유전자의 공배양과 이 정자의 난자내 직접도입법에 의해 외래유전자를 가진 형질전환 소 수정란과 형질전환 소 생산이 가능할 것으로 생각된다.

  • PDF

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
    • /
    • 제35권2호
    • /
    • pp.137-141
    • /
    • 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).

양측 엉덩이의 거대 신경섬유종 (Giant Neurofibroma on Both Buttocks)

  • 김지훈;범진식;김양우;강소라;김형경
    • Archives of Plastic Surgery
    • /
    • 제36권4호
    • /
    • pp.512-515
    • /
    • 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.

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

  • 김형석;정지열;강상철;신보문;이은주;황의경;박봉균;김대용;김재훈
    • 한국임상수의학회지
    • /
    • 제28권2호
    • /
    • pp.232-235
    • /
    • 2011
  • Clostridium (C.) difficile 은 사람뿐만 아니라 동물에서도 새롭게 출현한 중요한 병원체로 인식되어 왔다. 제주도에서 설사 증상이 있는 132두의 돼지 직장 분변 및 냉동 결장을 대상으로 C. difficile 균체의 독소 A, B 유전자를 검출하기 위한 중합효소연쇄반응을 실시하였다. 그 결과 포유자돈 2두 이유자돈 2두 및 육성단계 돼지 1두 등 총 5두의 돼지에서 독소 A 및 B에 대하여 양성반응을 나타내었다. PCR 결과는 돼지 대장의 조직학적 병변과 일치하였다. 병리조직학적 소견으로 독소 양성 돼지의 맹장과 결장에서는 점막하직 및 장막의 부종이 관찰되었다. 점막의 병변은 국소적인 상피세포의 괴사, 부분적인 탈락 및 장벽의 괴사까지 다양하였다. 세균 검사 결과 4두 (80%)는 Salmonella typhimurium과 혼합감염되어 있었다.

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

  • 김원일;박상열;김상진;조용일;허태영;김남수
    • 한국임상수의학회지
    • /
    • 제30권4호
    • /
    • pp.223-229
    • /
    • 2013
  • 송아지는 초유를 통해서만 모체항체를 받을 수 있기 때문에 신생우가 분만초기에 초유를 섭취하는 것은 질병에 대한 저항성을 갖추기 위해 필수적이다. 따라서 분만 후 잉여 초유를 저장하였다가 초유섭취가 부족한 신생우에 공급하여 충분한 항체를 갖도록 하는 것이 일반적인 사양법이다. 본 연구에서는 국내 3곳의 지역에서 운영되는 초유은행에서 저장된 35개의 초유를 무작위로 수거하여 일반세균오염과 주요 병원체들을 검사하였고 면역글로불린의 농도와 BVDV 특이 항체가를 측정하여 초유의 면역학적 품질을 평가하였다. 초유은행에서 수거된 대부분의 초유에서 중등도에서 고도의 세균오염이 관찰되었다. 또한 대부분의 초유가 좋은 품질의 초유라고 판단되는 50 g/L의 IgG 농도에 미치지 못하는 IgG를 포함하고 있었다. 따라서 초유은행 보관용 초유는 청결한 채취법에 따라 분만 후 2-3회 이하로 착유한 초유를 사용하여 안전성과 품질을 보장하여야 할 것이다. 또한 $63^{\circ}C$에서 30분간 실시하는 저온살균이 면역글로불린의 파괴를 최소화하는 동시에 초유에 의한 질병전파를 확연히 줄여주는 것으로 관찰되었으므로 냉동보관 전에 초유의 저온살균 과정을 반드시 실시하는 것이 필요할 것으로 판단된다.

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

  • 김해철;양성렬;배세진;심재형
    • 한국해양학회지:바다
    • /
    • 제3권2호
    • /
    • pp.80-89
    • /
    • 1998
  • 남극 내만 생태계에 서식하는 식물플랑크톤의 계절에 따른 분포 및 생리상태, 그리고 상호관계를 알아보기 위하여 아남극에 위치한 Maxwell 만과 Marian 소만에서 1994년 2월부터 1994년 12월까지 물리, 화학적 환경요인과 식물플랑크톤의 색소량, 일차생산력을 조사하였다. 조사시기 동안 관측한 chlorophyll a의 농도는 미검출 수준(not detected)~3.03 ${\mu}g/l$ (평균 0.63 ${\mu}g/l$)의 비교적 큰 변화를 보였고, 일차생산력은 0.53~18.95 mg C/$m^3{\cdot}day$를 기록하였으며, 수층적분한 식물플랑크톤의 일차생산력은 41.28~560.20 mg C/$m^3{\cdot}day$의 범위를 보였다. 식물플랑크톤의 생물량은 일사량과 양의 상관관계를 보였고($r^2$=0.29, p < 0.01), 일차생산력은 일사량과 매우 밀접한 관련이 있었으나($r^2$=0.85, P < 0.001), 온도와 영양염은 식물플랑크톤 군집을 조절하는데 기여하지 않았다.

  • PDF

Polarization Precession Effects for Shear Elastic Waves in Rotated Solids

  • Sarapuloff, Sergii A.
    • 한국소음진동공학회:학술대회논문집
    • /
    • 한국소음진동공학회 2013년도 춘계학술대회 논문집
    • /
    • pp.842-848
    • /
    • 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.

  • PDF

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

  • 주석천;이병천;이원유;최윤석;황우석
    • 대한수의학회지
    • /
    • 제37권3호
    • /
    • pp.647-659
    • /
    • 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.

  • PDF

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

  • 장용근;김권천
    • 대한두경부종양학회지
    • /
    • 제15권1호
    • /
    • pp.70-75
    • /
    • 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.

  • PDF