• Title/Summary/Keyword: Fusion rates

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Production of Clone Animals by Nuclear Transplantation I. Effects of Electrostimulation on Membrane Fusion of Embryos and Activation of Oocytes in Mouse (핵치환에 의한 Clone Animal의 생산에 관한 연구 I. 생쥐 수정란의 세포막 융합과 난모세포의 활성화에 미치는 전기자극의 효과)

  • 이상진;구덕본;이상민;박흠대;정순영;정길생
    • Korean Journal of Animal Reproduction
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    • v.18 no.3
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    • pp.217-228
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    • 1994
  • These experiments were carried out to establish the optimal condition of electrostimulatin inducing cell fusion and oocyte activation for nuclear transplantation in mouse embryos. Eggs selected for cell fusion or activation by electrostimulation were equilibrated for 5~10 min. in 0.3M sucrose solution and electrostimulated for 60$\mu$sec using 1 pulse of 60, 70, 80, 90 or 100 volts DC with electrodes 0.2 mm apart. Then they were cultured in 20${mu}ell$ dropsof Tyrode's solution. The results of these experiments are as follows : 1. When one pulse of 60, 70, 80, 90 or 100 volts DC for 60$\mu$sec were applied to 2-cell embryos for fusion of blastomeres, fusion rates were 50.0, 81.7, 91.7, 100 and 100%, respectively ; and developmental rates of fused embryos to blastocyst were 76.7 to 81.5%. Higher fusion rates were observed in 90V and 100V. 2. The average cell number in fused embryos developed to blastocyst was about half of the cell number in diploid controls; and the cell number decreased with increasing of voltages. 3. When pulse numbers were increased, fusion rates improved, but developmental rates were not signficiantly different from the group for which the number of pulse was not increased. And the cell number of blastocyst decreased even more. 4. Oocytes aged for 6hrs after ovulation were electrostimulated for oocyte activation by the same method used for cell fusion. Rates of oocyte activated by electrostimulation were 45.3 to 60.4%, and fragmentation rates were 7.5~15.1%. The lysis rates were 17.0~34.0%. The results of these experiments indicate that the optimal condition for achieving cell fusion and activation is 1 pulse, duration 60$\mu$sec in 90 Volt. The results also show that this condition is suitable for nuclear transplantation using mouse eggs.

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Blastomeres Fusion of 2-cell Mouse Embryos by Electric Stimulation (전기자극법에 의한 생쥐 2세포기란의 분할구 융합에 관한 연구)

  • Yang, B.Y.;Han, S.I.;Kim, C.I.
    • Clinical and Experimental Reproductive Medicine
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    • v.18 no.1
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    • pp.35-40
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    • 1991
  • To find out the suitable method for blastomeres fusion of mouse 2-cell embryo using electric stimuli, these studies were carried out with various voltages (1.0 KV, 1.2 KV, 1.5 KV, 1.7 KV and 2.0KV), pulse duration times($50{\mu}\;sec$, $75/{\mu}\;sec$, $100{\mu}\;sec$) and different fusion solutions. In addition, the fused embryos were cultured for 72-80hr to observe their subsequent development. These results were summarized as follows: 1. The proportion of the fused embryos were 50.8%(34/67), 60.7%(34/56), 70.6%(48/68), 66.7% (48/72) and 85.3% (58/68) after stimuli of 1.0KV, 1.2KV, 1.5KV, 1.7KV and 2.0KV for $100{\mu}\;sec$ with 2 times, and the electric stimulation at 2.0KV(85.3%) was the most effective voltage on the blastomere fusion. 2. For in vitro development, blastocysts of the fused embryos were cultured for 72-80hrs in $M_{16}$ medium. The group(52.1%) treated with 1.5KV for $100{\mu}\;sec$ with 2 times showd higher development rates than those any other group. However, these results were not corresponded to those of the rates of blastomere fusion. 3. There were no significant differences among the rates of blastomeres fusion to 50(70.6%), 75(71.9%), and 100(78.0%) ${\mu}sec$ stimulation at 1.5KV with two times. However, the development rates of the fused embryo in vitro were 52.1%(25/48), 28.3%(13/46) and 9.4%(3/32) at the above conditions, and the development rates of fused embryo increased as the pulse duration times increased. 4. The rates of the blastomeres fusion were 38.9% (28/72) or 70.6% (48/68) in electrolyte (PBS) or non-electrolyte(0.3M mannitol) solution. The development rates of the fused embryo were 32.1% (9/28) or 52.1%(25/48) in the above fusion solutions, and non-electrolyte-treated group showed higher development rates of embryo than that of electrolyte-treated group.

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Induction of twinning in Korean native cattle by transfer of nuclear transplanted embryos I. Embryo cloning using ovum pick-up(OPU), electric cell fusion and in vitro culture system (핵이식 기법을 이용한 한우 쌍태생산에 관한 연구 I. Ovum pick-up(OPU), 전기적 세포융합 및 체외배양 기법을 이용한 복제수정란 생산)

  • Hwang, Woo-suk;Shin, Tae-young;Roh, Sang-ho;Lee, Byeong-chun
    • Korean Journal of Veterinary Research
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    • v.38 no.1
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    • pp.209-216
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    • 1998
  • The purpose of present study is to improve the efficiency of fusion and the developmental rates of nuclear transplanted embryos to produce genetically identical twins from Korean native cattle. The diameter of aspirated follicles had no significant effect on the recovery rates of oocytes collected by ovum pick-up technique. The fusion rates of nuclear transplanted embryos were significantly higher in 50 and $100{\mu}s$ DC duration groups(73.3 and 72.0% ; respectively) than that in $30{\mu}s$ group(55.6% ; p<0.05). The cleavage rates of nuclear transplanted embryos appeared to be significantly higher in donor nuclei derived from in vivo (65.0%) than in those from in vitro (50.5% ; p<0.01), but the developmental rates to morulae and blastocysts were not significantly different between them(13.7 vs 10.9%, respectively).

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Do Trunk Muscles Affect the Lumbar Interbody Fusion Rate? : Correlation of Trunk Muscle Cross Sectional Area and Fusion Rates after Posterior Lumbar Interbody Fusion Using Stand-Alone Cage

  • Choi, Man Kyu;Kim, Sung Bum;Park, Bong Jin;Park, Chang Kyu;Kim, Sung Min
    • Journal of Korean Neurosurgical Society
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    • v.59 no.3
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    • pp.276-281
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    • 2016
  • Objective : Although trunk muscles in the lumbar spine preserve spinal stability and motility, little is known about the relationship between trunk muscles and spinal fusion rate. The aim of the present study is to evaluate the correlation between trunk muscles cross sectional area (MCSA) and fusion rate after posterior lumbar interbody fusion (PLIF) using stand-alone cages. Methods : A total of 89 adult patients with degenerative lumbar disease who were performed PLIF using stand-alone cages at L4-5 were included in this study. The cross-sectional area of the psoas major (PS), erector spinae (ES), and multifidus (MF) muscles were quantitatively evaluated by preoperative lumbar magnetic resonance imaging at the L3-4, L4-5, and L5-S1 segments, and bone union was evaluated by dynamic lumbar X-rays. Results : Of the 89 patients, 68 had bone union and 21 did not. The MCSAs at all segments in both groups were significantly different (p<0.05) for the PS muscle, those at L3-4 and L4-5 segments between groups were significantly different (p=0.048, 0.021) for the ES and MF muscles. In the multivariate analysis, differences in the PS MCSA at the L4-5 and L5-S1 segments remained significant (p=0.048, 0.043 and odds ratio=1.098, 1.169). In comparison analysis between male and female patients, most MCSAs of male patients were larger than female's. Fusion rates of male patients (80.7%) were higher than female's (68.8%), too. Conclusion : For PLIF surgery, PS muscle function appears to be an important factor for bone union and preventing back muscle injury is essential for better fusion rate.

Studies on the culture of bovine nuclear transplant embryos derived in vitro fertilization (체외수정 유래 소 핵이식란의 배양에 관한 연구)

  • Hwang, Woo-suk;Jo, Choong-ho
    • Korean Journal of Veterinary Research
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    • v.35 no.1
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    • pp.179-185
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    • 1995
  • The fusion rates of nuclear transplant embryos with various DC voltage were 55.6-79.2%. The significantly higher fusion rates of nuclear transplant embryos were achieved at the electric field strenght of DC 1.0-2.0kV/cm(72.0-79.2%) than DC 0.75kV/cm(55.6%, P<0.05). No significant differences in the percentage of embryos that cleaved(48.1, 55.4 and 42.6% respectively) and the percentage of cleaved embryos that developed to morulae/blastocyst(1.9, 5.3 and 1.9% respectively) could be found among the three types of in vitro culture system (Granulosa cell, BOEC co-culture and SOF, P>0.01). The age of the recipient cytoplast(30 vs 40hr) had no effect on the fusion rates and the rates of cleavage development(36.9 vs 44.1%, P>0.01).

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Optimization of Electrofusion Condition for the Production of Korean Cattle Somatic Cell Nuclear Transfer Embryos

  • Kim, Se-Woong;Kim, Dae-Hwan;Jung, Yeon-Gil;Roh, Sang-Ho
    • Reproductive and Developmental Biology
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    • v.35 no.1
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    • pp.17-22
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    • 2011
  • This study was designed to determine the effect of electric field strength, duration and fusion buffer in fusion parameters on the rate of membrane fusion between the somatic cell and cytoplast for Korean cattle (HanWoo) somatic cell nuclear transfer (SCNT) procedure. Following electrofusion, effect of 5 or $10\;{\mu}M$ $Ca^{2+}$-ionophore of activation treatment on subsequent development was also evaluated. Cell fusion rates were significantly increased from 23.1% at 20 V/mm to 59.7% at 26 V/mm and 52.9% at 27 V/mm (p<0.05). Due to higher cytoplasmic membrane rupture or cellular lysis, overall efficiency was decreased when the strength was increased to 30 V/mm (18.5%) and 40 V/mm (6.3%) and the fusion rate was also decreased when the strength was at 25 V/mm or below. The optimal duration of electric stimulation was significantly higher in $25\;{\mu}s$ than 20 and $30\;{\mu}s$ (18.5% versus 9.3% and 6.3%, respectively, p<0.05). Two nonelectrolyte fusion buffers, Zimmermann's (0.28 M sucrose) and 0.28 M mannitol solution for cell fusion, were used for donor cell and ooplast fusion and the fusion rate was significantly higher in Zimmermann's cell fusion buffer than in 0.28 M mannitol (91.1% versus 48.4%, respectively, p<0.05). The cleavage and blastocyst formation rates of SCNT bovine embryos activated by $5\;{\mu}M$ $Ca^{2+}$-ionophore was significantly higher than the rates of the embryos activated with $10\;{\mu}M$ of $Ca^{2+}$-ionophore (70.0% versus 42.9% and 22.5% versus 14.3%, respectively; p<0.05). This result is the reverse to that of parthenotes which shows significantly higher cleavage and blastocyst rates in $10\;{\mu}M$ $Ca^{2+}$-ionophore than $5\;{\mu}M$ counterpart (65.6% versus 40.3% and 19.5% versus 9.7%, respectively; p<0.05). In conclusion, SCNT couplet fusion by single pulse of 26 V/mm for $25\;{\mu}s$ in Zimmermann's fusion buffer followed by artificial activation with $5\;{\mu}M$ $Ca^{2+}$-ionophore are suggested as optimal fusion and activation methods in Korean cattle SCNT protocol.

Lumbar Interbody Fusion Outcomes in Degenerative Lumbar Disease : Comparison of Results between Patients Over and Under 65 Years of Age

  • Jo, Dae-Jean;Jun, Jae-Kyun;Kim, Ki-Tack;Kim, Sung-Min
    • Journal of Korean Neurosurgical Society
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    • v.48 no.5
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    • pp.412-418
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    • 2010
  • Objective : To evaluate the clinical and radiological outcomes of lumbar interbody fusion and its correlation with various factors (e.g., age, comorbidities, fusion level, bone quality) in patients over and under 65 years of age who underwent lumbar fusion surgery for degenerative lumbar disease. Methods : One-hundred-thirty-three patients with lumbar degenerative disease underwent lumbar fusion surgery between June 2006 and June 2007 and were followed for more than one year. Forty-eight (361%) were older than 65 years of age (group A) and 85 (63.9%) were under 65 years of age (group B). Diagnosis, comorbidities, length of hospital stay, and perioperative complications were recorded. The analysis of clinical outcomes was based on the visual analogue scale (VAS). Radiological results were evaluated using plain radiographs. Clinical outcomes, radiological outcomes, length of hospital stay, and complication rates were analyzed in relation to lumbar fusion level, the number of comorbidities, bone mineral density (BMD), and age. Results : The mean age of the patients was 61.2 years (range, 33-86 years) and the mean BMD was -2.2 (range, -4.8 to -2.8). The mean length of hospital stay was 15.0 days (range, 5-60 days) and the mean follow-up was 23.0 months (range, 18-30 months). Eighty-five (64.0%) patients had more than one preoperative comorbidities. Perioperative complications occurred in 27 of 133 patients (20.3%). The incidence of overall complication was 22.9% in group A, and 18.8% in group B but there was no statistical difference between the two groups. The mean VAS scores for the back and leg were significantly decreased in both groups (p < 0.05), and bony fusion was achieved in 125 of 133 patients (94.0%). There was no significant difference in bony union rates between groups A and B (91.7% in group A vs. 95.3% in group B, p = 0.398). In group A. perioperative complications were more common with the increase in fusion level (p = 0.027). Perioperative complications in both groups A (p = 0.035) and B (p = 0.044) increased with an increasing number of comorbidities. Conclusion : Elderly patients with comorbidities are at a high risk for complications and adverse outcomes after lumbar spine surgery. In our study, clinical outcomes, fusion rates, and perioperative complication rates in older patients were comparable with those in younger populations. The number of comorbidities and the extent of fusion level were significant factors in predicting the occurrence of postoperative complications. However, proper perioperative general supportive care with a thorough fusion strategy during the operation could improve the overall postoperative outcomes in lumbar fusion surgery for elderly patients.

Fusion Between TMPRSS2 and ETS Family Members (ERG, ETV1, ETV4) in Prostate Cancers from Northern China

  • Wang, Jian-Jiang;Liu, Yue-Xin;Wang, Wei;Yan, Wei;Zheng, Yu-Peng;Qiao, Lu-Dong;Liu, Dan;Chen, Shan
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.10
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    • pp.4935-4938
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    • 2012
  • In this study we evaluated the frequency of fusion between TMPRSS2 and ETS family members (ERG, ETV1, ETV4) in prostate cancers in patients from northern China in order to explore differences in fusion rates among regions in northern and southern China, other parts of Asia, Europe, and North America. We examined 100 prostate cancer patients, diagnosed by means of prostate biopsy; fluorescence in situ hybridization (FISH) was used to detect the expression of TMPRSS2, ERG, ETV1 and ETV4 in cancer tissue. Differences in gene fusion rates among different ethnics groups were also analyzed. Of the 100 prostate cancer patients, 55 (55%) had the fusion gene. Among the patients with the fusion gene, 46 (83.6%) patients had the TMPRSS2:ERG fusion product, 8 (14.8%) patients had TMPRSS2:ETV1 fusion, 1 (1.6%) patient had TMPRSS2:ETV4.

Pulsed Electromagnetic Field Stimulators Efficacy for Noninvasive Bone Growth in Spine Surgery

  • Fiani, Brian;Kondilis, Athanasios;Runnels, Juliana;Rippe, Preston;Davati, Cyrus
    • Journal of Korean Neurosurgical Society
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    • v.64 no.4
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    • pp.486-494
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    • 2021
  • The growth of pulsed electromagnetic field (PEMF) therapy and its progress over the years for use in post-operative bone growth has been revolutionary in its effect on bone tissue proliferation and vascular flow. However, further progress in PEMF therapy has been difficult due to lack of more evidence-based understanding of its mechanism of action. Our objective was to review the current understanding of bone growth physiology, the mechanism of PEMF therapy action along with its application in spinal surgery and associated outcomes. The authors of this review examined multiple controlled, comparative, and cohort studies to compare fusion rates of patients undergoing PEMF stimulation. Examining spinal fusion rates, a rounded comparison of post-fusion outcomes with and without bone stimulator was performed. Results showed that postoperative spinal surgery PEMF stimulation had higher rates of fusion than control groups. Though PEMF therapy was proven more effective, multiple factors contributed to difficulty in patient compliance for use. Extended timeframe of treatment and cost of treatment were the main obstacles to full compliance. This review showed that PEMF therapy presented an increased rate of recovery in patients, supporting the use of these devices as an effective post-surgical aid. Given the recent advances in the development of PEMF devices, affordability and access will be much easier suited to the patient population, allowing for more readily available treatment options.

Production of Cloned Embryos by Nuclei Transfer and Electronic Cell Fusion from Bovine Fetal Fibroblasts (Bovine Fetal Fibroblasts를 이용한 핵이식 및 세포융합에 관한 연구)

  • 이병천;박종임;조종기;김기연;신수정;용환율;황우석
    • Journal of Embryo Transfer
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    • v.14 no.2
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    • pp.107-111
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    • 1999
  • The present study was performed to evaluate the best electric fusion condition in nuclear transfer, Korean Native Cattle fibroblasts were used as nucleic donors. Oocytes from slaughterhouse were matured in vitro for 22 h and enucleated. Each individual cells were transferred into enucleated ocytes and reconstructed embryo were placed into the fusion chamber. In experiment 1, pulse were performed by altering pulse duration at 1. 75kv/cm, 1 time. When pulse duration is 30$mutextrm{s}$, fusion and development rates is higher than other conditions. In experiment 2, the effect of different pulse number were studied at the pulse duration 30$mutextrm{s}$ and the same pulse intensity. When pulse number was one, fusion rates were higher than other conditions. The fused embryos were moved to culture medium and assessed their development to blastocyst. These results showed that best fusion condition was 30$mutextrm{s}$ and one time. And the fibroblasts derived from Han Woo can be reprogrammed by nuclear transplantation and develop subsequently in vitro.

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