• Title/Summary/Keyword: Air-to-Air

Search Result 35,088, Processing Time 0.077 seconds

THE EFFECTS OF VARIOUS TETRACYCLINE HCL CONCENTRATION TREATED ROOW SURFACES ON PROLIFERATION AND SPREADING OF PERIODONTAL LIGAMENT CELLS (다양한 농도의 테트라사이클린로 처리된 치근면이 치주인대세포의 증식과 전개에 미치는 영향)

  • Jung, Oh-Chul;Sun, Jo-Young
    • Journal of Periodontal and Implant Science
    • /
    • v.24 no.3
    • /
    • pp.581-596
    • /
    • 1994
  • This in vitro study was undertaken to obtain optimal tetracycline concentration that aids proliferation and spreading of human periodontal ligament cells, for clinical application in root surfaces of periodontally diseased teeth. Periodontal ligament cells used in this study were obtained from explants of periodontal ligament of 1st premolar teeth which were extracted for the purpose of orthodontic treatment. The cells were cultured in Dulbecco's Modified Eagle Medium(DMEM) supplemented with 100 U/ml penicillin, $100\;{\mu}g/ml$ streptomycin and 10% FBS at $37^{\circ}C$, 100% humidity, 5% $CO_2-95%$ air. Cells were used between the third to 4th passage. After root planing of periodontally extracted teeth, the root slabs were cut with carborundum disk. In the cell proliferation experiment, experimental groups were root planing only group, immersed groups in 25, 50, 75, 100, 150mg/ml aqueous solution of Tetracycline HCl followed by a vigorous rinse in PBS. Human PDL cells at concentration of $1{\times}10^5\;cells/ml$ were seeded in each culture well which contained root slabs and incubated for 6 hours. Then, all of the root slabs were moved into new 24 culture well and incubated 24, 48 and 72 hours. The cell counting was done by inverted phase contrast microscope after trypsinization. The following results were obtained. The cell number was increased in order root planing only group, 25, 150, 50, 75, 100mg/ml of Tetracycline HCl treated group in 24, 48 and 72 hours. The maximal cell number was obtained when the root slabs were immersed in solution with 100mg/ml of Tetracycline HCl. There were statistically significant between the root planing only group and 75, 100 mg/ml of Tetracycline HCl treated group in 24 hours, between the root planing only group and 100mg/ml of Tetracycline HCl treated group in 48 hours, between the root planing only group and 50, 75, 100mg/ml of Tetracycline HCl treated group, between 25 and 100mg/ml of Tetracycline HCl treated group in 72 hours(p<0.05). In the cell spreading experiment, after 30 minutes of incubated, in the root planing only group, the cells were generally round in shape. The cell surface was mostly covered with blebs. The cells started to attach to root surface by cytoplasmic extension in 50, 100mg/ml of Tetracycline HCl treated groups, more numerous cells attached to root surface than root planing only group. Many orifices of dentinal tubule were exposed, cells showed radially spreaded cytoplasm and unspreaded central region of the cell was covered with blebs. After 6 hours of incubation, in the root planing only group, cells showed radially spreaded cytoplasm and were attached flat appearance. In 50, 100mg/ml of Tetracycline HCl treated groups, cellular margin was concaved and cytoplasm showed elongated appearance with polarity. After 24 hours of incubation, in the root planing group, cells showed characteristic polarity. In 50, 100mg/ml of Tetracycline HCl treated groups, cells showed more elongated and spindle - like appearance.

  • PDF

A study on dermatologic diseases of workers exposed to cutting oil (절삭유 취급 근로자의 피부질환에 관한 연구)

  • Chun, Byung-Chul;Kim, Hee-Ok;Kim, Soon-Duck;Oh, Chil-Hwan;Yum, Yong-Tae
    • Journal of Preventive Medicine and Public Health
    • /
    • v.29 no.4 s.55
    • /
    • pp.785-799
    • /
    • 1996
  • We investigated the 1,004 workers who worked in a automobile factory to study the epidemiologic characterists of dermatoses due to cutting oils. Among the workers, 667(66.4%) answered the questionaire. They are belong to 5 departments of the factory-the Engine-Work(258 workers), Gasoline engine Assembly(210), Diesel engine Assembly(96), Power train Work(86), Power train Assembly(17). We measured the oil mist concentration in air of the departments and examined the workers who had dermatologic symptoms. The results were follows; 1) Oil mist concentration ; Of all measured points(52),9 points(17.2%) exeeded $5mg/m^3$- the time-weighed PEL-and one department had a upper confidence limit(95%) higher than $5mg/m^3$. 2) Dermatologists examined 213 workers. 172 of them complained any skin symptoms at that time - itching(32.5%), papule(21.6%), scale(15.7%), vesicle(12.5%) in order. The abnormal skin site found by dermatologist were palm(29.3%), finger & nail(24.6%), forearm(16.2%), back of hand(8.4%) in order. 3) As the result of physical examination, we found that 160 workers had skin diseases. Contact dermatitis was the most common; 69 workers had contact dermatitis alone(43.1%), 11 had contact dermatitis with acne(6.9%), 10 had contact dermatitis with folliculitis(6.3%), 1 had contact dermatitis with acne & folliculitis, and 1 had contact dermatitis with abnormal pigmentation. Others were folliculitis(9 workers, 5.6%), acne(8, 5.0%), folliculitis & acne (2, 1.2%), keratosis(1, 0.6%), abnormal pigmentation (1, 0.6%), and non-specific hand eczema (47, 29.3%). 4) The prevalence of any skin diseases was 34.0 pet 100 in cutting oil users, and 13.3 per 100 in non- users. Especially, the prevalence of contact dermatitis was 23.0 per 100 in cutting oil users and 23.0 per 100 in non-users. 5) We tried patch test(standard serise, oil serise, organic solvents) on 49 patients to differentiate allergic contact dermatitis from irritant contact dermatitis and found 20 were positive. 6) In a multivariate analysis(independant=age, tenure, kinds of cutting oil), the risk of skin diseases was higher in the water-based cutting oil user and both oil user than non-user or neat oil user(odds ratio were 2.16 and 2.78, respectively). And the risk of contact dermatitis was much higher at the same groups(odds ratio were 5.16 and 6.82, respectively).

  • PDF

Effects of Soil Organic Matter Contents, Paddy Types and Agricultural Climatic Zone on CH4 Emissions from Rice Paddy Field (벼 논에서 토양 유기물 함량, 논 유형 및 농업기후대가 CH4 배출에 미치는 영향)

  • Ko, Jee-Yeon;Lee, Jae-Saeng;Woo, Koan-Sik;Song, Seok-Bo;Kang, Jong-Rae;Seo, Myung-Chul;Kwak, Do-Yeon;Oh, Byeong-Gun;Nam, Min-Hee
    • Korean Journal of Soil Science and Fertilizer
    • /
    • v.44 no.5
    • /
    • pp.887-894
    • /
    • 2011
  • To evaluate the effects of abiotic factors of paddy fields on greenhouse gases (GHGs) emissions from rice paddy fields, $CH_4$ emission amounts were investigated from rice paddy fields by different soil organic matter contents, paddy types, and agricultural climatic zone in Yeongnam area during 3 years. $CH_4$ emission amounts according to soil organic matter contents in paddy field were conducted at having different contents of 5 soil organic matters fields (23.6, 28.7, 31.0, 34.5, and $38.0g\;kg^{-1}$), The highest $CH_4$ emission amount was recorded in the highest soil organic matters plot of $38.0g\;kg^{-1}$. High correlation coefficient (r=$0.963^{**}$) was obtained between $CH_4$ emissions from paddy fields and their soil organic matter contents. According to paddy field types, $CH_4$ emission amounts were investigated at 4 different paddy fields as wet paddy, sandy paddy, immature paddy, and mature paddy. The highest $CH_4$ emissions was recorded in wet paddy (100%) and followed as immature paddy 64.0%, mature paddy 46.8%, and sandy paddy 23.8%, respectively. For the effects of temperature on $CH_4$ emissions from paddy fields, 4 agricultural climatic zones were investigated, which were Yeongnam inland zone (YIZ), eastern coast of central zone (ECZ), plain area of Yeongnam inland mountainous zone (PMZ), and mountainous area of Yeongnam inland mountainous zone (MMZ). The order of $CH_4$ emission amounts from paddy fields by agricultural climatic zone were YIZ (100%) > ECZ (94.6%) > PMZ (91.6%) > MMZ (78.9%). The regression equation between $CH_4$ emission amounts from paddy fields and average air temperature of Jul. to Sep. of agricultural climatic zone was y = 389.7x-4,287 (x means average temperature of Jul. to Sep. of agricultural climatic zone, $R^2=0.906^*$)

Effects of Fertilization Time and Culture Medium of Pig Oocytes Matured In Vitro by liquid Boar Sperm Stored at $4^{\circ}C$ (체외성숙된 돼지난포란을 $4^{\circ}C$ 보존 액상정액으로 체외수정시 수정시간과 배양배지의 영향)

  • Park, C. S.;Y. J. Yi;Kim, M. Y.;Y. J. Chang;Lee, S. H.;D. I. Jin
    • Korean Journal of Animal Reproduction
    • /
    • v.27 no.3
    • /
    • pp.215-223
    • /
    • 2003
  • This study was to investigate the effects of fertilization time and culture medium of pig oocytes matured in-vitro by liquid boar sperm. The sperm rich fraction (30∼60 ml) was slowly cooled to room temperature (20∼23$^{\circ}C$) by 2 h after collection. Semen was transferred into 15 ml tubes, centrifuged at room temperature for 10 min 800 ${\times}$ g, and the supernatant solution was poured off. The concentrated sperm was resuspended with 5 ml of the LEN diluent to provide 1.0${\times}$10$^{9}$ sperm/ml at room temperature. The resuspended semen was cooled in a refrigerator to 4$^{\circ}C$. The medium used for oocyte maturation was TCM-199 supplemented with 26.19 mM sodium bicarbonate, 0.9 mM sodium pyruvate, 10 $\mu\textrm{g}$/ml insulin, 2 $\mu\textrm{g}$/ml vitamin B$_{12}$ , 25 mM HEPES, 10 $\mu\textrm{g}$/ml bovine apotransferrin, 150 $\mu$M cysteamine, 10 IU/ml PMSG, 10 IU/ml hCG, 10 ng/ml EGF, 0.4% BSA, 75 $\mu\textrm{g}$/ml sodium penicillin G, 50 $\mu\textrm{g}$/ml streptomycin sulfate and 10% pFF. After about 22 h of culture, oocytes were cultured without cysteamine and hormones for 22 h at 38.5$^{\circ}C$, 5% $CO_2$ in air. Oocytes were inseminated with liquid boar sperm stored at 4$^{\circ}C$ for 2 days after collection. Oocytes were coincubated for 1, 3, 6 and 9 h in 500 ${mu}ell$ mTBM fertilization media with 1.0${\times}$10$^{6}$ sperm/ml concentration, respectively. Thereafter, oocytes were transferred into 500 ${mu}ell$ NCSU-23, HEPES buffered NCSU-23, PZM-3 and PZM-4 culture media, respectively, for further culture of 6, 48 and 144 h. The rates of sperm penetration and male pronuclear formation were higher in the fertilization times for 6 and 9 h than in those for 1 and 3 h. The rates of cleaved oocytes were higher in the fertilization times for 6 and 9 h (85.0 and 84.6%) than in those for 1 and 3 h (61.1 and 76.8%). The percentage of blastocyst formation from the cleaved oocytes was highest in the fertilization time for 6 h (33.6%) than in that for 1, 3 and 9 h (11.4, 23.0 and 29.6%). Mean cell numbers per blastocyst were 32.9, 27.6, 26.3 and 24.4 in the fertilization times for 6, 9, 3 and 1 h, respectively. The rate of blastocyst from the cleaved oocytes and the number of cells per blastocyst were higher in HEPES buffered NCSU-23 culture medium than in NCSU-23, PZM-3 and PZM-4 culture media. In conclusion, we found out that liquid boar sperm stored at 4$^{\circ}C$ could be used for in-vitro fertilization of pig oocytes matured in-vitro. Also, we recommend the coincubation time of 6 h in 500 ${mu}ell$ TBM fertilization medium with 1${\times}$10$^{6}$ sperm/ml concentration and the HEPES buffered NCSU-23 culture medium for in-vitro fertilization of pig oocytes matured in-vitro.

Experimental Study of Flip-Bucket Type Hydraulic Energy Dissipator on Steep slope Channel (긴구배수로 감세공의 Filp Bucket형 이용연구)

  • 김영배
    • Magazine of the Korean Society of Agricultural Engineers
    • /
    • v.13 no.1
    • /
    • pp.2206-2217
    • /
    • 1971
  • Spillway and discharge channel of reservoirs require the Control of Large volume of water under high pressure. The energies at the downstream end of spillway or discharge channel are tremendous. Therefore, Some means of expending the energy of the high-velocity flow is required to prevent scour of the riverbed, minimize erosion, and prevent undermining structures or dam it self. This may be accomplished by Constructing an energy dissipator at the downstream end of spillway or discharge channel disigned to dissipated the excessive energy and establish safe flow Condition in the outlet channel. There are many types of energy dissipators, stilling basins are the most familar energy dissipator. In the stilling basin, most energies are dissipated by hydraulic jump. stilling basins have some length to cover hydraulic jump length. So stilling basins require much concrete works and high construction cost. Flip bucket type energy dissipators require less construction cost. If the streambed is composed of firm rock and it is certain that the scour will not progress upstream to the extent that the safety of the structure might be endangered, flip backet type energy dissipators are the most recommendable one. Following items are tested and studied with bucket radius, $R=7h_2$,(medium of $4h_2{\geqq}R{\geqq}10h_2$). 1. Allowable upstream channel slop of bucket. 2. Adequate bucket lip angle for good performance of flip bucket. Also followings are reviwed. 1. Scour by jet flow. 2. Negative pressure distribution and air movement below nappe flow. From the test and study, following results were obtained. 1. Upstream channel slope of bucket (S=H/L) should be 0.25<H/L<0.75 for good performance of flip bucket. 2. Adequated lip angle $30^{\circ}{\sim}40^{\circ}$ are more reliable than $20^{\circ}{\sim}30^{\circ}$ for the safety of structures.

  • PDF

A study of usefulness for the plan based on only MRI using ViewRay MRIdian system (ViewRay MRIdian System을 이용한 MRI only based plan의 유용성 고찰)

  • Jeon, Chang Woo;Lee, Ho Jin;An, Beom Seok;Kim, Chan young;Lee, Je hee
    • The Journal of Korean Society for Radiation Therapy
    • /
    • v.27 no.2
    • /
    • pp.131-143
    • /
    • 2015
  • Purpose : By comparing a CT fusion plan based on MRI with a plan based on only MRI without CT, we intended to study usefulness of a plan based on only MRI. And furthermore, we intended to realize a realtime MR-IGRT by MRI image without CT scan during the course of simulation, treatment planning, and radiation treatment. Materials and Methods : BBB CT (Brilliance Big Bore CT, 16slice, Philips), Viewray MRIdian system (Viewray, USA) were used for CT & MR simulation and Treatment plan of 11 patients (1 Head and Neck, 5 Breast, 1 Lung, 3 Liver, 1 Prostate). When scanning for treatment, Free Breathing was enacted for Head&Neck, Breast, Prostate and Inhalation Breathing Holding for Lung and Liver. Considering the difference of size between CT and Viewray, the patient's position and devices were in the same condition. Using Viewray MRIdian system, two treatment plans were established. The one was CT fusion treatment plan based on MR image. Another was MR treatment plan including electron density that [ICRU 46] recommend for Lung, Air and Bone. For Head&Neck, Breast and Prostate, IMRT was established and for Lung and Liver, Gating treatment plan was established. PTV's Homogeneity Index(HI) and Conformity Index(CI) were use to estimate the treatment plan. And DVH and dose difference of each PTV and OAR were compared to estimate the treatment plan. Results : Between the two treatment plan, each difference of PTV's HI value is 0.089% (Head&Neck), 0.26% (Breast), 0.67% (Lung), 0.2% (Liver), 0.4% (Prostate) and in case of CI, 0.043% (Head&Neck), 0.84% (Breast), 0.68% (Lung), 0.46% (Liver), 0.3% (Prostate). As showed above, it is on Head&Neck that HI and CI's difference value is smallest. Each difference of average dose on PTV is 0.07 Gy (Head&Neck), 0.29 Gy (Breast), 0.18 Gy (Lung), 0.3 Gy (Liver), 0.18 Gy (Prostate). And by percentage, it is 0.06% (Head&Neck), 0.7% (Breast), 0.29% (Lung), 0.69% (Liver), 0.44% (Prostate). Likewise, All is under 1%. In Head&Neck, average dose difference of each OAR is 0.01~0.12 Gy, 0.04~0.06 Gy in Breast, 0.01~0.21 Gy in Lung, 0.06~0.27 Gy in Liver and 0.02~0.23 Gy in Prostate. Conclusion : PTV's HI, CI dose difference on the Treatment plan using MR image is under 1% and OAR's dose difference is maximum 0.89 Gy as heterogeneous tissue increases when comparing with that fused CT image. Besides, It characterizes excellent contrast in soft tissue. So, radiation therapy using only MR image without CT scan is useful in the part like Head&Neck, partial breast and prostate cancer which has a little difference of heterogeneity.

  • PDF

In Vitro Fertilization of Pig Oocytes Matured In­Vitro by liquid Boar Spermatozoa (체외성숙 돼지 난포란의 액상정액을 이용한 체외수정)

  • 박창식;이영주
    • Korean Journal of Animal Reproduction
    • /
    • v.26 no.1
    • /
    • pp.17-23
    • /
    • 2002
  • The present study was carried out to investigate the effects of the maturation media such as a modified TCM-199 (mTCM-199) medium, modified Waymouth MB 752/1 (mWaymouth MB 752/1) medium or NCSU-23 medium on penetrability of pig oocytes by liquid boar sperm. Oocytes (30~40) were transferred into each well of a Nunc 4-well multidish containing 0.5 $m\ell$ maturation medium. When immature pig oocytes were cultured in mTCM-199, mWaymouth MB 752/1 and NCSU-23 maturation media for 44 h in 5% $CO_2$, in air at 38.5$^{\circ}C$, the germinal vesicle breakdown (CVBD) rates of the oocytes were 95.6, 94.1 and 94.9%, respectively, and the maturation rates (metaphase II) of oocytes were 92.5, 90.1 and 91.1%, respectively. No differences were observed among the maturation media. The spermrich portion of ejaculates with greater than 90% motile sperm were used in the experiment. The semen was cooled 22 to 24$^{\circ}C$ over 2 h period. The semen was diluted with Beltsville Thawing Solution (BTS) extender at room temperature to give 2$\times$10$^{8}$ sperm/$m\ell$ in 100 $m\ell$ plastic bottle. Liquid boar semen of 30 $m\ell$ in 100 $m\ell$ plastic bottle was kept at 17$^{\circ}C$ for 5 days. The sperm with greater than 70% motility after day 5 of storage were used for in-vitro fertilization (IVF). After 44 h maturation of immature oocytes, cumulus cells were removed and oocytes (30~40) coincubated far 6 h in 0.5 $m\ell$ mTCM-199 and mTBM fertilization media with 2$\times$1061$m\ell$ sperm concentration. At 6 h after IVF, oocytes were transferred into 0.5 $m\ell$ mTCM-199 and NCSU-23 culture media for further culture 6 or 42 h. Sperm penetration, polyspermy and male pronuclear formation of oocytes at 12 h after IVF, and developmental ability of oocytes at 48 h after IVF were evaluated. The oocytes in combination with NCSU-23 medium for maturation and mTBM medium for IVF increased male pronuclear formation (48.0%) compared to those in combination with mTCM-199 media for maturation and IVF, and mWaymouth MB 752il medium for maturation and mTCM-199 medium far IVF. The rates of cleaved embryos (2~4 cell stage) at 48 h after IVF were 24.1% in combination with mTCM-199 media for maturation, IVF and culture, 43.6% in combination with mWaymouth MB 75211 medium fur maturation and mTCM-199 media for IVF and culture, and 71.2% in combination with NCSU-23 medium for maturation, mTBM medium for IVF and NCSU-23 medium for culture. In conclusion, we found out the oocytes matured in vitro were fertilized by liquid boar sperm stored in BTS extender at 17$^{\circ}C$ for 5 days. We recommend the simple defined NCSU-23 medium for nuclear maturation, mTBM medium and liquid boar sperm for IVF, and NCSU-23 medium for embryo culture.

Determination of Optimal Concentration of LPE (Lysophosphatidylethanolamine) for Postharvest Stability and Quality of Strawberry Fruit (딸기 수확 후 저장기간 연장 및 품질 개선을 위한 LPE (Lysophosphatidylethanolamine) 적정 처리농도 구명)

  • Choi, Ki-Young;Kim, Il-Seop;Yun, Young-Sik;Choi, Eun-Young
    • Journal of Bio-Environment Control
    • /
    • v.25 no.3
    • /
    • pp.153-161
    • /
    • 2016
  • This study aims to determine the optimal maturity of strawberry fruits as affected by the application of lysophosphatidylethanolamine (LPE) and its optimal concentration for postharvest stability and quality. Prior to application of treatments, fruits that were classified into levels of maturity (0%, 50%, 70% and 100%) were air-dried for 40 minutes and stored in the refrigerator at $4^{\circ}C$ for 12 days. Fruits at 70% maturity were dipped into 0, 10, 50 and $100mg{\cdot}L^{-1}$ LPE solutions for 1 minute. A lower range of concentration (0, 2.5, 5, 10 and $25mg{\cdot}L^{-1}$) was applied to fruits at different maturity levels. Data on fresh weight, hardness at vertical and horizontal loading positions, color index and sugar content during storage were collected. Based on fruits with 70% maturity dipped in LPE concentrations, there were no significant differences found on fresh weight, color index and sugar content. However, the application of $10mg{\cdot}L^{-1}$ LPE gave the highest hardness at vertical loading position while $100mg{\cdot}L^{-1}$ had the lowest average. At lower range of LPE concentrations, fresh weight was not significantly affected by LPE application and maturity levels. Hardness of fruits was mainly based on the maturity of the fruits. Increased hardness was observed in the fruits with 70% maturity dipped into the $5mg{\cdot}L^{-1}$ of LPE solution. The hardness and Hunter's $L^*$ and $b^*$ value of 100% matured fruits gave lowest values despite the application of $25mg{\cdot}L^{-1}$ LPE 12 days after storage.

Studies on the fate of nitrogen in the paddy soil (답토양(沓土壤)에서 질소(窒素)의 동태(動態)에 관(關)한 연구(硏究))

  • Kim, Kwang Sik
    • Korean Journal of Soil Science and Fertilizer
    • /
    • v.9 no.1
    • /
    • pp.17-23
    • /
    • 1976
  • In order to investigate the fate of nitrogen in the paddy soil, Suchang, Hwasoon and Susan soil which have different properties, were treated with several nitrogen fertilizers such as ammonium chloride, ammonium sulfate, urea and SCU (sulfur-coated urea), and incubated under water-logged condition in $30^{\circ}C$ incubator. $NH_4-N$, $NO_3-N$, $Fe^{++}$ and pH in soil and stagnant water, were determined at 10, 20, 30, 40 and 50 days after incubation. The obtained results were summarized as follows: 1. The effect of rising temperature was increased in order of Hwasoon>Suchang>Susan and the effect of air drying soil was risen in order of Susan>Hwasoon>Suchang, while the rate of ammonication was in order of Susan>Suchang>Hwasoon. 2. The changes of $NH_4-N$ in stagnant water was dependent upon the nitrogen concentration of $NH_4Cl$ and $(NH_4)SO_4$ plat was high and decreased after 30 days incubation, but increased after 40 days and then decreased again. In contrast with the above, $NH_4-N$ concentration of urea and SCU plot was low but the change showed slightly through the incubation period. 3. Accumulation of $NH_4-N$ in the oxidative layer of the $NH_4Cl$ and $(NH_4)_2SO_4$ plot was higher than that of urea and SCU plot and $NH_4-N$ content was decreased with the incubation period. The change of $NH_4-N$ in the reductive layer showed the same pattern. 4. The changes of $NO_3-N$ in the stagnant water were different according to soil properties and nitrogen fertilizer. $NO_3-N$ concentration in stagnant water of urea and SCU plot was higher than in the $NH_4-Cl$ $(NH_4)_2SO_4$ plot and nearly disappeared after 30 to 40 days incubation. 5. The $NO_3-N$ concentration in the oxidative layer of soil was higher than reductive layer. The pattern of change was different in accordance with soil properties and nitrogen fertilizers. In general, nitrification in urea and SCU plot was more increased than $(NH_4)_2SO_4$ plot. In reductive layer, the concentration of $NO_3-N$ was very low until 30 days incubation and thereafter increased slightly. 6. Upon the concentration of $NH_4-N$ and $NO_3-N$ in stagnant water and soil, it was assumed that denitification of urea and SCU plot was higher than $NH_4Cl$ and $(NH_4)_2SO_4$ plot and denitrified nitrogen in incubation period was above 50%.

  • PDF

Effects of Recipient Oocytes and Electric Stimulation Condition on In Vitro Development of Cloned Embryos after Interspecies Nuclear Transfer with Caprine Somatic Cell (수핵난자와 전기적 융합조건이 산양의 이종간 복제수정란의 체외발달에 미치는 영향)

  • 이명열;박희성
    • Reproductive and Developmental Biology
    • /
    • v.28 no.1
    • /
    • pp.21-27
    • /
    • 2004
  • This study was conducted to investigate the developmental ability of caprine embryos after somatic cell interspecies nuclear transfer. Recipient bovine and porcine oocytes were obtained from slaughterhouse and were matured in vitro according to established protocols. Donor cells were obtained from an ear-skin biopsy of a caprine, digested with 0.25% trypsin-EDTA in PBS and primary fibroblast cultures were established in TCM-199 with 10% FBS. The matured oocytes were dipped in D-PBS plus 10% FBS + 7.5 $\mu$ g/ml cytochalasin B and 0.05M sucrose. Enucleation were accomplished by aspirating the first polar body and partial cytoplasm which containing metaphase II chromosomes using a micropipette with an out diameter of 20∼30 $\mu$m. A Single donor cell was individually transferred into the perivitelline space of each enucleated oocyte. The reconstructed oocytes were electric fusion with 0.3M mannitol fusion medium. After the electrofusion, embryos were activated by electric stimulation. Interspecies nuclear transfer embryos with bovine cytoplasts were cultured in TCM-199 medium supplemented with 10% FBS including bovine oviduct epithelial cells for 7∼9 day. And porcine cytoplasts were cultured in NCSU-23 medium supplemented with 10% FBS for 6 ∼8 day at $39^{\circ}C, 5% CO_2 $in air. Interspecies nuclear transfer by recipient bovine oocytes were fused with electric length 1.95 kv/cm and 2.10 kv/cm. There was no significant difference between two electric length in fusion rate(47.7 and 44.6%) and in cleavage rate(41.9 and 54.5%). Using electric length 1.95 kv/cm and 2.10 kv/cm in caprine-porcine NT oocytes, there was also no significant difference between two treatments in fusion rate(51.3 and 46.1%) and in cleavage rate(75.0 and 84.9%). The caprine-bovine NT oocytes fusion rate was lower(P<0.05) in 1 pulse for 60 $\mu$sec(19.3%), than those from 1 pulse for 30 $\mu$sec(50.8%) and 2 pulse for 30 $\mu$sec(31.0%). The cleavage rate was higher(P<0.05) in 1 pulse for 30 $\mu$sec(53.3%) and 2 pulse for 30 $\mu$sec(50.0%), than in 1 pulse for 60 $\mu$sec(18.2%). The caprine-porcine NT oocytes fusion rate was 48.1% in 1 pulse for 30 $\mu$sec, 45.2% in 2 pulse for 30 $\mu$sec and 48.6% in 1 pulse for 60 $\mu$sec. The cleavage rate was higher(P<0.05) in 1 pulse for 30 $\mu$sec(78.4%) and 1 pulse for 60 $\mu$sec(79.4%), than in 2 pulse for 30 $\mu$sec(53.6%). In caprine-bovine NT embryos, the developmental rate of morula and blastocyst stage embryos were 22.6% in interspecies nuclear transfer and 30.6% in parthenotes, which was no significant differed. The developmental rate of morula and blastocyst stage embryos with caprine-porcine NT embryos were lower(P<0.05) in interspecies nuclear transfer(5.1%) than parthenotes(37.4%).