• Title/Summary/Keyword: CO2 laser

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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
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    • v.28 no.1
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    • pp.21-27
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    • 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%).

High Resolution HC$_3$N Observations toward the Central Region of Sagittarius B2

  • H.S-Ching;Oh, M.ishi;M.Morimoto
    • Bulletin of the Korean Space Science Society
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    • 1993.10a
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    • pp.17-17
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    • 1993
  • The wall shear stress in the vicinity of end-to end anastomoses under steady flow conditions was measured using a flush-mounted hot-film anemometer(FMHFA) probe. The experimental measurements were in good agreement with numerical results except in flow with low Reynolds numbers. The wall shear stress increased proximal to the anastomosis in flow from the Penrose tubing (simulating an artery) to the PTFE: graft. In flow from the PTFE graft to the Penrose tubing, low wall shear stress was observed distal to the anastomosis. Abnormal distributions of wall shear stress in the vicinity of the anastomosis, resulting from the compliance mismatch between the graft and the host artery, might be an important factor of ANFH formation and the graft failure. The present study suggests a correlation between regions of the low wall shear stress and the development of anastomotic neointimal fibrous hyperplasia(ANPH) in end-to-end anastomoses. 30523 T00401030523 ^x Air pressure decay(APD) rate and ultrafiltration rate(UFR) tests were performed on new and saline rinsed dialyzers as well as those roused in patients several times. C-DAK 4000 (Cordis Dow) and CF IS-11 (Baxter Travenol) reused dialyzers obtained from the dialysis clinic were used in the present study. The new dialyzers exhibited a relatively flat APD, whereas saline rinsed and reused dialyzers showed considerable amount of decay. C-DAH dialyzers had a larger APD(11.70${\pm}$1.32mmHg/min)compared to CF dialyzers(4.32${\pm}$0.55mmHg/min)(p<0.05). However, there was no observable difference in the UFR between the two dialyzers. Neither APD nor UFR showed any significant increase with an increasing number of reuses for up to more than 20reuses. A substantial number of failures observed in APD(larger than 20mmHe/min)on the reused dialyzers(2 out of 40 CP and S out 26 C-DAK) were attributed to the Possible damage on the fibers. The CF 15-11 HFDs which failed APD test did not show changes in the UFR compared to normal dialyzers indicating that APD is a more sensitive test than UFR test to evaluate the integrity of the fibers. 30527 T00401030527 ^x For quantitative measurement of reflected light from a clinical diagnostic strip, a prototype old reflectance photometer was designed. The strip loader and cassette were made to obtain more accurate reflectance parameters. The strip was illuminated at 45˚c through optical fiber and the intensity of reflected light was determined at rectanguLat angle using a photodiode. The kubelka-munk coefficient and reflection optical density were determined ar four different wavelengths(500,550,570 and 610nm) for blood glucose strip. For higher concentration than 300mg/41 about glucose, a saturation state of abforbance was observed at 500,550 and 570nm. The correlation between glucose concentration and parameters was the best at 610nm. 30535 T00401030535 ^x Radiation-induced fibrosarcoma tumors were grown on the flanks of C3H mice. The mice were divided into two groups. One group was injected with Photofrin II, intravenously (2.5mg/kg body weight). The other group received no Photofrin II. Mice from both groups were irradialed for approximately 15 minutes at 100,300, or 500 mW/cm2 with the argon (488nm/514.5 nm), dye(628nm) and gold vapor (pulsed 628 nm) laser light. A photosensitizer behaved as an added absorber. Under our experimental conditions, the presence of Photolfrin II increased surface temperature by at least 40% and the temperature rise due to 300 mW/cm2 irradiation exceeded values for hyperthermia. Light and temperature distributions with depth were estimated by a computer model. The model demonstrated the influence of wavelength on the thermal process and proved to be a valuable tool to investigate internal temperature rise. 30536 T00401030536 ^x We investigated the structural geometry of thirty-eight Korean femurs. The purpose of this study is to identify major geometrical differences between Korean femurs 3nd others that we believe belong to Caucasians so that we would be able to get insights into the femoral component design that fits Asians including Koreans. We utilized computerized tomography (CT) images of femurs extracted from cadavers. The CT images were transformed into bitmap data by using a film scanner, and then analyzed by using a commercially available software called Image v.1.0 and a Macintosh IIci computer.The resulting data were compared with already published data. The major results show that the geometry of the Korean femurs is significantly different from that of Caucasians: (1) the anteversion angle and the canal flare index are greater by the amount of approximately 8˚ and 0.5, respectively, (2) the shape of the isthmus cross section is more round, and (3) the distance between the teaser trochanter and the proximal border of the isthmus is shelter by about 15 mm. The results suggested that the femoral component suitable for Asians should be different from the currently-used components designed and manufactured mostly by European or American companies. 30537 T00401030537 ^x It is well known that nonlinear propagation characteristics of the wave in the tissue may give very useful information for the medical diagnoisis. In this paper, a new method to detect nonlinear propagation characteristics of the internal vibration in the tissue for the low frequency mechanical vibration by using bispectral analysis is proposed. In the method, low frequency vibration of f0( = 100Hz) is applied on the surface of the object, and the waveform of the internal vibration x (t) is measured from Doppler frequency modulation of silmultaneously transmitted probing ultrasonic waves. Then, the bispectra of the signal x (t) at the frequencies (f0, f0) and (f0, 2f0) are calculated to estimate the nonlinear propagation characteristics as their magnitude ratio, w here since bispectrum is free from the gaussian additive noise we can get the value with high S/N. Basic experimental system is constructed by using 3.0 MHz probing ultrasonic waves and the several experiments are carried out for some phantoms. Results show the superiority of the proposed method to the conventional method using power spectrum and also its usefulness for the tissue characterization. 30541 T00401030541 ^x This paper describes the implementation of a computerized radial pulse diagnosis by aids of a clinical expert. On this base, we composed of the radial pulse diagnosis system in korean traditional medicine. The system composed of a radial pulse wave detection system and a radial pulse diagnosis system. With a detection system, we detected Inyoung and Cheongu radial pulse wave and processed it. Then, we have got the characteristic parameters of radial pulse wave and also quantified that according to the method of Inyoung-Cheongu Comparison Radial Pulse Diagnosis. We defined the jugement standard of radial pulse diagnosis system and then we confirmed the possibility for realization of automatic radial pulse diagnosis in korean traditional medicine. 30545 T00401030545 ^x Microspheres are expected to be applied to biomedical areas such as solid-phase immunoassays, drug delivery systems, immunomagnetic cell separation. To synthesize microspheres for biomedical application, "two stage shot growth method" was developed. The uniformity ratio of synthesized microspheres was always smaller than 1.05. And the surface charge density (or the number of ionizable functional groups) of the microspheres synthesized by "two stage shot growth method" was 6~13 times higher than that of the microspheres synthesized by conventional seeded batch copolymerization. As a previous step for biomedical application, adsorption experiments of bovine albumin on microspheres were carried out under various conditions. The maximum adsorbed amount was obtained in the neighborhood of pH 4.5. Isoelectric point of bovine albumin is pH 5.0, so experimental result shows that it shifted to acid area. The adsorption isotherm was obtained, the plateau region was always reached at 2.Og/L (bulk concentration of bovine albumin).The effect of the kind and the amount of surface functional group was also examined. 30575 T00401030575 ^x A medical image workstation was developed using multimedia technique. The system based on PC-486DX was designed to acquire medical images produced by medical imaging instruments and related audio information, that is, doctors' reporting results. Input information was processed and analyzed, then the results were presented in the form of graph and animation. All the informations of the system were hierarchically related with the image as the apex. Processing and analysis algorithms were implemented so that the diagnostic accuracy could be improved. The diagnosed information can be transferred for patient diagnosis through LAN(local area network). 30592 T00401030592 ^x In the conventional infrared imaging system, complex infrared lens systems are usually used for directing collimated narrow infrared beams into the high speed 2-dimensional optic scanner. In this paper, a simple reflective infrared optic system with a 2-dimensional optic scanner is proposed for the realization of medical infrared thermography system. It has been experimentally proven that the intfrared thermography system composed of the proposed optic system has the temperature resolution of 0.1˚c under the spatial resolution of lmrad, the image matrix size of 256 X 240, and tile imaging time of 4 seconds. 30593 T00401030593 ^x In this paper, MIIS (Medical Image Information System) has been designed and implemented using INGRES RDBMS, which is based on a client/server architecture. The implemented system allows users to register and retrieve patient information, medical images and diagnostic reports. It also provides the function to display these information on workstation windows simultaneously by using the designed menu-driven graphic user interface. The medical image compression/decompression techniques are implemented and integrated into the medical image database system for the efficient data storage and the fast access through the network. 30594 T00401030594 ^x In this paper, computerized BEAM was implemented for the space domain analysis of EEG. Trans-formation from temporal summation to two-dimensional mappings is formed by 4 nearest point inter-polaton method. Methods of representation of BEAM are two. One is dot density method which classify brain electrical potential 9 levels by dot density of gray levels and the other is colour method which classify brain electrical 12 levels by red-green colours. In this BEAM, instantaneous change and average energy distribution over any arbitrary time interval of brain electrical activity could be observed and analyzed easily. In the frequency domain, the distribution of energy spectrum of a special band can easily be distinguished normality and abnormality. 30608 T00401030608 ^x Laboratory information system (LIS) is a key tool to manage laboratory data in clinical pathology. Our department has developed an information system for routine hematology using down-sized computer system. We have used an IBM 486 compatible PC with 16MB main memory, 210 MB hard disk drive, 9 RS-232C port and 24 pin dot printer. The operating system and database management system were SCO UNIX and SCO foxbase, respectively. For program development, we used Xbase language provided by SCO foxbase. The C language was used for interface purpose. To make the system use friendly, pull-down menu was used. The system connected to our hospital information system via application program interface (API), so the information related to patient and request details is automatically transmitted to our computer. Our system interfaced with fwd complete blood count analyzers(Sysmex NE-8000 and Coulter STKS) for unidirectional data tansmission from analyzer to computer. The authors suggests that this system based on down-sized computer could provide a progressive approach to total LIS based on local area network, and the implemented system could serve as a model for other hospital's LIS for routine hematology. 30609 T00401030609 ^x To develop an artificial bone substitute that is gradually degraded and replaced by the regenerated natural bone, the authors designed a composite that is consisted of calcium phosphate and collagen. To use as the structural matrix of the composite, collagen was purified from human umbilical cord. The obtained collagen was treated by pepsin to remove telopeptides, and finally, the immune-free atelocollagen was produced: The cross linked atelocollagen was highly resistant to the collagenase induced collagenolysis. The cross linked collagen demonstrated an improved tensile strength. 30618 T00401030618 ^x This paper is a study on the design of adptive filter for QRS complex detection. We propose a simple adaptive algorithm to increase capability of noise cancelation in QRS complex detection with two stage adaptive filter. At the first stage, background noise is removed and at the next stage, only spectrum of QRS complex components is passed. Two adaptive filters can afford to keep track of the changes of both noise and QRS complex. Each adaptive filter consists of prediction error filter and FIR filter The impulse response of FIR filter uses coefficients of prediction error filter. The detection rates for 105 and 108 of MIT/BIH data base were 99.3% and 97.4% respectively. 30619 T00401030619 ^x To develop an artificial bone substitute that is gradually degraded and replaced by the regenerated natural bone, the authors designed and produced a composite that is consisted of calcium phosphate and collagen. Human umbilical cord origin pepsin treated type I atelocollagen was used as the structural matrix, by which sintered or non-sintered carbonate apatite was encapsulated to form an inorganic-organic composite. With cross linking atelocollagen by UV ray irradiation, the resistance to both compressive and tensile strength was increased. Collagen degradation by the collagenase induced collagenolysis was also decreased. 30620 T00401030620 ^x We have developed a monoleaflet polymer valve as an inexpensive and viable alternative, especially for short-term use in the ventricular assist device or total artificial heart. The frame and leaflet of the polymer valve were made from polyurethane, To evaluate the hemodynamic performance of the polymer valve a comparative study of flow dynamics past a polymer valve and a St. Jude Medical prosthetic valve under physiological pulsatile flow conditions in vitro was made. Comparisons between the valves were made on the transvalvular pressure drop, regurgitation volume and maximum valve opening area. The polymer valve showed smaller regurgitation volume and transvalvular pressure drop compared to the mechanical valve at higher heart rate. The results showed that the functional characteristics of the polymer valve compared favorably with those of the mechanical valve at higher heart rate. 30621 T00401030621 ^x Explosive evaporative removal process of biological tissue by absorption of a CW laser has been simulated by using gelatin and a multimode Nd:YAG laser. Because the point of maximun temperature of laser-irradiated gelatin exists below the surface due to surface cooling, evaporation at the boiling temperature is made explosively from below the surface. The important parameters of this process are the conduction loss to laser power absorption (defined as the conduction-to-laser power parameter, Nk), the convection heat transfer at the surface to conduction loss (defined as Bi), dimensionless extinction coefficient (defined as Br.), and dimensionless irradiation time (defined as Fo). Dependence of Fo on Nk and Bi has been observed by experiment, and the results have been compared with the numerical results obtained by solving a 2-dimensional conduction equation. Fo and explosion depth (from the surface to the point of maximun temperature) are increased when Nk and Bi are increased.To find out the minimum laser power for explosive evaporative removal process, steady state analysis has been also made. The limit of Nk to induce evaporative removal, which is proportional to the inverse of the laser power, has been obtained. 30622 T00401030622 ^x N1 and N2 gross neural action potentials were measured from the round window of the guinea pig cochlea at the onset of the acoustic stimuli. N1-N2 audiograms were made by means of regulating stimulant intensities in order to produce constant N1-N2 potentials as criteria for different input tone pip frequencies. The lowest threshold was measured with an input tone pip I5 dB SPL in intensity and 12 KHz in frequency when the animal was in normal physiological condition. The procedure of experimental measurements is explained in detail. This experimental approach is very useful for the investigation of the Cochlear function. Both noN1inear and active functions of the Cochlea can be monitored by N1-N2 audiograms. 30623 T00401030623 ^x In electrical impedance tomography(EIT), we use boundary current and voltage measurements toprovide the information about the cross-sectional distribution of electrical impedance or resistivity. One of the major problems in EIT has been the inaccessibility of internal voltage or current data in finding the internal impedance values. We propose a new image reconstruction method using internal current density data measured by NMR. We obtained a two-dimensional current density distribution within a phantom by processing the real and imaginary MR images from a 4.77 NMR machine. We implemented a resistivity mage reconstruction algorithm using the finite element method and sensitivity matrix. We presented computer simulation results of the mage reconstruction algorithm and furture direction of the research. 30624 T00401030624 ^x A new method of digital image analysis technique for discrimination of cancer cell was presented in this paper. The object image was the Thyroid eland cells image that was diagnosed as normal and abnormal (two types of abnormal: follicular neoplastic cell, and papillary neoplastic cell), respectively. By using the proposed region segmentation algorithm, the cells were segmented into nucleus. The 16 feature parameters were used to calculate the features of each nucleus. A9 a consequence of using dominant feature parameters method proposed in this paper, discrimination rate of 91.11% was obtained for Thyroid Gland cells. 30625 T00401030625 ^x An electrical stimulator was designed to induce locomotion for paraplegic patients caused by central nervous system injury. Optimal stimulus parameters, which can minimize muscle fatigue and can achieve effective muscle contraction were determined in slow and fast muscles in Sprague-Dawley rats. Stimulus patterns of our stimulator were designed to simulate electromyographic activity monitored during locomotion of normal subjects. Muscle types of the lower extremity were classified according to their mechanical property of contraction, which are slow muscle (msoleus m.) and fast muscle (medial gastrocneminus m., rectus femoris m., vastus lateralis m.). Optimal parameters of electrical stimulation for slow muscles were 20 Hz, 0.2 ms square pulse. For fast muscle, 40 Hz, 0.3 ms square pulse was optimal to produce repeated contraction. Higher stimulus intensity was required when synergistic muscles were stimulated simultaneously than when they were stimulated individually. Electrical stimulation for each muscle was designed to generate bipedal locomotion, so that individual muscles alternate contraction and relaxation to simulate stance and swing phases. Portable electrical stimulator with 16 channels built in microprocessor was constructed and applied to paraplegic patients due to lumbar cord injury. The electrical stimulator restored partially gait function in paraplegic patients. 30626 T00401030626 ^x Two-Dimensional modelling of the Cochlear biomechanics is presented in this paper. The Laplace partial differential equation which represents the fluid mechanics of the Cochlea has been transformed into two-dimensional electrical transmission line. The procedure of this transformation is explained in detail. The comparison between one and two dimensional models is also presented. This electrical modelling of the basilar membrane (BM) is clearly useful for the next approach to the further. Development of active elements which are essential in the producing of the sharp tuning of the BM. This paper shows that two-dimension model is qualitatively better than one-dimensional model both in amplitude and phase responses of the BM displacement. The present model is only for frequency response. However because the model is electrical, the two-dimensional transmission line model can be extended to time response without any difficult. 30627 T00401030627 ^x A method has been proposed for the fully automatic detection of left ventricular endocardial boundary in 2D short axis echocardiogram using geometric model. The procedure has the following three distinct stages. First, the initial center is estimated by the initial center estimation algorithm which is applied to decimated image. Second, the center estimation algorithm is applied to original image and then best-fit elliptic model estimation is processed. Third, best-fit boundary is detected by the cost function which is based on the best-fit elliptic model. The proposed method shows effective result without manual intervention by a human operator. 30628 T00401030628 ^x The intelligent trajectory control method that controls moving direction and average velocity for a prosthetic arm is proposed by pattern recognition and force estimations using EMG signals. Also, we propose the real time trajectory planning method which generates continuous accelleration paths using 3 stage linear filters to minimize the impact to human body induced by arm motions and to reduce the muscle fatigue. We use combination of MLP and fuzzy filter for pattern recognition to estimate the direction of a muscle and Hogan's method for the force estimation. EMG signals are acquired by using a amputation simulator and 2 dimensional joystick motion. The simulation results of proposed prosthetic arm control system using the EMf signals show that the arm is effectively followed the desired trajectory depended on estimated force and direction of muscle movements. 30638 T00401030638 ^x A new neural network architecture for the recognition of patterns from images is proposed, which is partially based on the results of physiological studies. The proposed network is composed of multi-layers and the nerve cells in each layer are connected by spatial filters which approximate receptive fields in optic nerve fields. In the proposed method, patterns recognition for complicated images is carried out using global features as well as local features such as lines and end-points. A new generating method of matched filers representing global features is proposed in this network. 30659 T00401030659 ^x An implementation scheme of the magnetic nerve stimulator using a switching mode power supply is proposed. By using a switching mode power supply rather than a conventional linear power supply for charging high voltage capacitors, the weight and size of the magnetic nerve stimulator can be considerably reduced. Maximum output voltage of the developed magnetic nerve stimulator using the switching mode power supply is 3,000 volts and switching time is about 100 msec. Experimental results or human nerve stimulations using the developed stimulator are presented. 30768 T00401030768 ^x In this paper, we describe the design methodology and specifications of the developed module-based bedside monitors for patient monitoring. The bedside monitor consists of a main unit and module cases with various parameter modules. The main unit includes a 12.1" TFT color LCD, a main CPU board, and peripherals such as a module controller, Ethernet LAN card, video card, rotate/push button controller, etc. The main unit can connect at maximum three module cases each of which can accommodate up to 7 parameter modules. They include the modules for electrocardiograph, respiration, invasive blood pressure, noninvasive blood pressure, temperature, and SpO2 with Plethysmograph.

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Re-Cloning by Somatic Cell Nuclear Transfer from a Cloned Korean Native Goat (복제 산양(진순이)의 체세포 핵이식에 의한 Re-Cloning에 관한 연구)

  • Jung, S.Y.;Park, H.S.
    • Journal of Embryo Transfer
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    • v.22 no.2
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    • pp.89-95
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    • 2007
  • The present study was conducted to examine some factors affecting in vitro development and fecundity of embryos recloned with somatic cell nuclear transfer (SCNT). Fibroblast cells retrieved from the ear of a 3-week-old, cloned Korean goat (Jinsoonny) were used as karyoplast donors and serum-starvation was conducted in tissue culture medium (TCM)-199 supplemented with 0.5% FBS. Recipient oocytes were surgically collected by flushing the oviducts 35 h after hCG injection following FSH priming. The zonae pellucidae of the oocytes were partially perforated with a laser drill and a donor cell was transferred into an enucleated oocyte. The couplets were electrically fused and activated by ionomycin (5 min) and 6-DMAP (4 h). The reconstructed embryos were cultured in mSOF medium containing 0.8% BSA at $39^{\circ}C$ in an atmosphere of 5% $CO_2$, 5% $%O_2$, 90% $N_2$ for 12 to 15 h. Re-cloned embryos (2- to 4-cell stages) were surgically transferred into the oviducts of the recipients and pregnancy was subsequently diagnosed by progesterone assay and ultrasound on Days 21 and 63 of pregnancy. The fusion rate following 1st fusion pulse was higher (p<0.05) in 2nd cloning (65.9%) compared to 1st cloning (51.0%), but it was not different in the other groups. The rate of cleavage after fusion was significantly higher (p<0.05) in 1st (77.7%) than in 2nd cloning (56.0%). A total of 175 re-cloned embryos were transferred into 28 recipients. On day 21 and 60 after transfer, 11 (39.3%) and 4 recipients (17.4%) were pregnancy, respectively. In comparison of pregnancy rate by estrous synchronization, a total of 66 and 109 re-cloned embryos were transferred into 11 recipients in natural estrus and 17 recipients in induced estrus, respectively. Five (45.4%) and 2 recipients (18.2%) in natural estrus were pregnant on days 21 and 63 while 6 (35.3%) and 2 (11.8%) recipients in induced estrus were pregnant, respectively. These results show that recloning of goat can be achieved by SCNT and estrous synchronization between donor and recipient animals may be one of the major factors affecting success rate.

Effects of Donor Cells and Estrus Synchronization on the Production of Cloned Korean Native Goat (공핵 세포 및 발정 동기화가 복제 재래 산양 생산에 미치는 영향)

  • Park H.S.;Kim T.S.;Jung S.Y.;Park J.K.;Lee J.S.;Jung J.Y.
    • Journal of Embryo Transfer
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    • v.21 no.2
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    • pp.137-146
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    • 2006
  • The objective of this study was to examine the effect of donor cell types, the source of recipient oocytes and estrous synchronization on pregnancy and delivery rates of somatic cell nuclear transfer (SCNT) embryos in Korean native goats. Recipient oocytes were surgically collected after superovulation. Ear cells and fetal fibroblasts were collected and cultured in serum-starvation condition (TCM-199 + 0.5% FBS) for cell confluence. The zonae pellucidae of in vivo- and in vitro-matured oocytes were partially drilled using a laser system. Single somatic cell was transferred into the enucleated oocyte. The reconstructed oocytes were electrically fused with 0.3 M mannitol. After the fusion, embryos were activated by Ionomycin+6-DMAP. NT embryos were cultured in mSOF medium supplemented with 0.8% BSA at $39^{\circ}C$ in an atmosphere of 5% $CO_2$, 5% $O_2$, 90% $N_2$ for 12 to 20 hr. One hundred and two SCNT embryos were transferred into 20 recipients and pregnancy rate at days 30 was 20.0%. Of them, one developed to term and delivered 1 kid. Ear cells showed significantly higher fusion (63.8 vs. 26.5%) and pregnancy rates (20.0 vs. 0.0%) than those of fetal fibroblast (p<0.05). The recipients synchronized by CIDR showed significantly lower pregnancy rates compared to that of recipient in natural estrus ($0.0{\sim}25.0%$ vs. 100%) (p<0.05). Cloned kid was born from the recipient in natural estrus. For the synchronization of estrus between recipient and donor, there was no difference between treatments (${\pm}0$ vs. +12 hr) in pregnancy rate. The first healthy cloned kid (Jinsoonny) was produced by transfer of SCNT embryos derived from in vivo oocytes and ear cells into a recipient goat whose estrus was synchronized with the donor. These results imply that donor cells for nuclear transfer may affect the success rate, and the estrus synchronization between donor and recipient animals can also be important.

Photodynamic Therapy with Photofrin Reduces Invasiveness of U87 Malignant Human Glioma Cells (교모세포종 세포주 U87에서 Photofrin을 사용한 광역학 치료가 종양 침습성에 미치는 영향)

  • Woo, Hye Kyung;Cho, Kyung-Keun;Rha, Hyung Kyun;Lee, Kyung Jin;Park, Sung Chan;Cho, Jung Ki;Park, Hea Kwan;Kang, Joon Ki;Choi, Chang Rak
    • Journal of Korean Neurosurgical Society
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    • v.30 no.sup2
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    • pp.189-196
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    • 2001
  • Objective : We tested the hypothesis that photodynamic therapy(PDT) with Photofrin inhibits tumor invasion of U87 human glioma cells using several in vitro assay to measure tumor invasiveness. The effects of PDT on cell growth, directional migration and cell invasion were investigated. Material and Method : Tumor cells were treated with Photofrin at various doses and at a fixed optical(632nm) dose of $100mJ/cm^2$. Cytotoxicity was tested using the MTT method. Invasion assays including the matrigelartificial basement membrane barrier migration and spheroid confrontation with confocal microscopic analysis were used to study the relationship between PDT and invasiveness. Result : U87 cells showed a dose dependent cytotoxic response to increasing Photofrin dose. Data from the matrigel artificial basement membrane assay indicate that PDT inhibits the U87 cell migration dose dependently. Low doses of subcytotoxic PDT treatment, such as 2.5ug/ml Photofrin dose, also appeared to significantly inhibit migration of U87 cells(p<0.05). In co-cultures between U87 cell spheroids and brain aggregates, progressive invasion with destruction of the brain aggregate occurs. The extent of tumor cell infiltration and proportion or intact brain aggregate remaining after 24h differs in Photofrin PDT treated versus Photofrin only control, with changes suggestive of a dose-response effect. Conclusion : our data indicate that PDT with Photofrin significantly inhibits the invasiveness of U87 cells, and this inhibition is dose dependent.

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Ginsenoside compound K protects against cerebral ischemia/ reperfusion injury via Mul1/Mfn2-mediated mitochondrial dynamics and bioenergy

  • Qingxia Huang;Jing Li;Jinjin Chen;Zepeng Zhang;Peng Xu;Hongyu Qi;Zhaoqiang Chen;Jiaqi Liu;Jing Lu;Mengqi Shi;Yibin Zhang;Ying Ma;Daqing Zhao;Xiangyan Li
    • Journal of Ginseng Research
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    • v.47 no.3
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    • pp.408-419
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    • 2023
  • Background: Ginsenoside compound K (CK), the main active metabolite in Panax ginseng, has shown good safety and bioavailability in clinical trials and exerts neuroprotective effects in cerebral ischemic stroke. However, its potential role in the prevention of cerebral ischemia/reperfusion (I/R) injury remains unclear. Our study aimed to investigate the molecular mechanism of ginsenoside CK against cerebral I/R injury. Methods: We used a combination of in vitro and in vivo models, including oxygen and glucose deprivation/reperfusion induced PC12 cell model and middle cerebral artery occlusion/reperfusion induced rat model, to mimic I/R injury. Intracellular oxygen consumption and extracellular acidification rate were analyzed by Seahorse multifunctional energy metabolism system; ATP production was detected by luciferase method. The number and size of mitochondria were analyzed by transmission electron microscopy and MitoTracker probe combined with confocal laser microscopy. The potential mechanisms of ginsenoside CK on mitochondrial dynamics and bioenergy were evaluated by RNA interference, pharmacological antagonism combined with co-immunoprecipitation analysis and phenotypic analysis. Results: Ginsenoside CK pretreatment could attenuate mitochondrial translocation of DRP1, mitophagy, mitochondrial apoptosis, and neuronal bioenergy imbalance against cerebral I/R injury in both in vitro and in vivo models. Our data also confirmed that ginsenoside CK administration could reduce the binding affinity of Mul1 and Mfn2 to inhibit the ubiquitination and degradation of Mfn2, thereby elevating the protein level of Mfn2 in cerebral I/R injury. Conclusion: These data provide evidence that ginsenoside CK may be a promising therapeutic agent against cerebral I/R injury via Mul1/Mfn2 mediated mitochondrial dynamics and bioenergy.

Studies on the In Vitro Development of Cloned Embryos by Somatic Cell Nuclear Transfer in Korean Native Goats (재래산양의 체세포 핵이식에 의한 복제수정란의 체외발달에 관한 연구)

  • Park H. S.;Kim T. S.;Jung S. Y.;Lee Y. H.;Jung J. Y.
    • Journal of Embryo Transfer
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    • v.20 no.2
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    • pp.105-112
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    • 2005
  • The present study was conducted to examine some factors affecting in vitro development of oocytes from somatic cell nuclear transfer (SCNT) in Korean native goats. Recipient oocytes were surgically collected after superovulation by using CIDR and FSH, PMSG, hCG and estrous synchronization in Korean Native goats. For nuclear transfer, the fibroblasts from caprine ear cells and fetal fibroblasts were surgically harvested and were cultured in vitro until cell confluency in serum-starvation condition (TCM-199 + $0.5\%$ FBS) for 3 to 5 days. The zona pellucidae of matured oocytes were partially drilled by laser irradiation. A single somatic cell was individually transferred into each enucleated oocyte. The reconstructed oocytes were then electrically fused and activated. Activated NT embryos were cultured in mSOF medium supplemented with $0.8\%\;BSA\;6\~7\;day\;at\;39^{\circ}C,\;5\%\;CO_2,\;5\%\;O_2,\;90\%\;N_2$ in air. There were no significant difference in the number of embryos cleaved and 4-cell development between the fibroblast nuclei from mature ear cells and fetal cells, but the rate of 8-cell development was higher (P<0.05) in ear cells $(40.5\%)$ than in fetal cells $(55.5\%)$. However, the embryo development to morula or blastocyst was not significantly different between both the groups$(6.7\%\;vs\;16.0\%)$, respectively. The number of embryo cleaved $(79.0\%)$ were higher (P<0.05) in the oocytes activated with ionomycin+6-DMAP than in the oocytes activated electrically $(9.5\%)$. The development of fused embryos to morula or blastocyst was found $15.6\%$ in ionomycin+6-DMAP, but no morula or blastocysts were developed in electrical stimulation. The development rate of SCNT embryos to morula or blastocyst was love. (P<0.05) in SCNT embryos $(19.0\%\;vs\;0.0\%)$ than that in parthenotes $(66.1\%\;vs\;59.1\%)$. In the parthenotes, the cleavage rate and development to morula or blastocyst were significantly higher (P<0.05) as $86.8\%\;and\;50.0\%$ in ovulated oocytes than in follicular oocytes $(69.0\%\;vs\;23.6\%)$, respectively. These results suggest that some factors Including superovulation treatment, oocyte source, maturation of follicular oocytes, activation method and culture condition may affect in vitro developmental capability of embryos produced by somatic cell nuclear transfer in Korean Native goats, and the fusion rate be greatly low compared with other species.

Stellite bearings for liquid Zn-/Al-Systems with advanced chemical and physical properties by Mechanical Alloying and Standard-PM-Route

  • Zoz, H.;Benz, H.U.;Huettebraeucker, K.;Furken, L.;Ren, H.;Reichardt, R.
    • Proceedings of the Korean Powder Metallurgy Institute Conference
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    • 2000.04a
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    • pp.9-10
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    • 2000
  • An important business-field of world-wide steel-industry is the coating of thin metal-sheets with zinc, zinc-aluminum and aluminum based materials. These products mostly go into automotive industry. in particular for the car-body. into building and construction industry as well as household appliances. Due to mass-production, the processing is done in large continuously operating plants where the mostly cold-rolled metal-strip as the substrate is handled in coils up to 40 tons unwind before and rolled up again after passing the processing plant which includes cleaning, annealing, hot-dip galvanizing / aluminizing and chemical treatment. In the liquid Zn, Zn-AI, AI-Zn and AI-Si bathes a combined action of corrosion and wear under high temperature and high stress onto the transfer components (rolls) accounts for major economic losses. Most critical here are the bearing systems of these rolls operating in the liquid system. Rolls in liquid system can not be avoided as they are needed to transfer the steel-strip into and out of the crucible. Since several years, ceramic roller bearings are tested here [1.2], however, in particular due to uncontrollable Slag-impurities within the hot bath [3], slide bearings are still expected to be of a higher potential [4]. The today's state of the art is the application of slide bearings based on Stellite\ulcorneragainst Stellite which is in general a 50-60 wt% Co-matrix with incorporated Cr- and W-carbides and other composites. Indeed Stellite is used as the bearing-material as of it's chemical properties (does not go into solution), the physical properties in particular with poor lubricating properties are not satisfying at all. To increase the Sliding behavior in the bearing system, about 0.15-0.2 wt% of lead has been added into the hot-bath in the past. Due to environmental regulations. this had to be reduced dramatically_ This together with the heavily increasing production rates expressed by increased velocity of the substrate-steel-band up to 200 m/min and increased tractate power up to 10 tons in modern plants. leads to life times of the bearings of a few up to several days only. To improve this situation. the Mechanical Alloying (MA) TeChnique [5.6.7.8] is used to prOduce advanced Stellite-based bearing materials. A lubricating phase is introduced into Stellite-powder-material by MA, the composite-powder-particles are coated by High Energy Milling (HEM) in order to produce bearing-bushes of approximately 12 kg by Sintering, Liquid Phase Sintering (LPS) and Hot Isostatic Pressing (HIP). The chemical and physical behavior of samples as well as the bearing systems in the hot galvanizing / aluminizing plant are discussed. DependenCies like lubricant material and composite, LPS-binder and composite, particle shape and PM-route with respect to achievable density. (temperature--) shock-reSistibility and corrosive-wear behavior will be described. The materials are characterized by particle size analysis (laser diffraction), scanning electron microscopy and X-ray diffraction. corrosive-wear behavior is determined using a special cylinder-in-bush apparatus (CIBA) as well as field-test in real production condition. Part I of this work describes the initial testing phase where different sample materials are produced, characterized, consolidated and tested in the CIBA under a common AI-Zn-system. The results are discussed and the material-system for the large components to be produced for the field test in real production condition is decided. Outlook: Part II of this work will describe the field test in a hot-dip-galvanizing/aluminizing plant of the mechanically alloyed bearing bushes under aluminum-rich liquid metal. Alter testing, the bushes will be characterized and obtained results with respect to wear. expected lifetime, surface roughness and infiltration will be discussed. Part III of this project will describe a second initial testing phase where the won results of part 1+11 will be transferred to the AI-Si system. Part IV of this project will describe the field test in a hot-dip-aluminizing plant of the mechanically alloyed bearing bushes under aluminum liquid metal. After testing. the bushes will be characterized and obtained results with respect to wear. expected lifetime, surface roughness and infiltration will be discussed.

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