• Title/Summary/Keyword: Pre-AMP

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Studies on In Vitro Fertilizability of Mouse Oocytes Pre-exposed to Dibutyryl Cyclic AMP (Dibutyryl Cyclic AMP로 처리된 생쥐난자의 수정능에 관한 연구)

  • 강해묵;이영기;조완규
    • The Korean Journal of Zoology
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    • v.31 no.1
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    • pp.21-28
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    • 1988
  • The present study was carried out to examine the fertilizability of the mouse oocytes pre-ex-posed to dbcAMP which is a well-known inhibitor of the oocyte maturation. The oocytes once cultured in the dbcMP-containing medium for a certain length of, time were cultivated in the dbcMp-free medium to induced the maturation, then mixed with sperms, and observed following culture for 24 hours. The fertilization rate of cocytes was judged by the index of the number of 2-cell embryo developed 24hr following insemination. The fertilization rate of the oocyte previously incubated with dbcAMP (100 g/ml) for 2, 4, 8 16 hours was 32.3, 14.5, 4.7 and 8.8%, respectively, while that of the control was 53.3% indicating that the fertilizability was decreased as a function of time exposed to dbcAMP. The pretreatment of dbcMP, however, didn't affect the process of sperm penetration to egg. In addition, there is no prominent changes in the morphological architecture of fertielized eggs which has been exposed to dbcAMP as revealed by electron microscopic observation. Consequendy, it can be concluded that the mouse cocytes once inhibited their maturation by dbcMP may retain, in some extent, the fertilizability, although most of the fertilized egg may not proceed to further development because of the failure of pronucleus formation.

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Comparative Analysis and Performance Evaluation of New Low-Power, Low-Noise, High-Speed CMOS LVDS I/O Circuits (저 전력, 저 잡음, 고속 CMOS LVDS I/O 회로에 대한 비교 분석 및 성능 평가)

  • Byun, Young-Yong;Kim, Tae-Woong;Kim, Sam-Dong;Hwang, In-Seok
    • Journal of the Institute of Electronics Engineers of Korea SC
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    • v.45 no.2
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    • pp.26-36
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    • 2008
  • Due to the differential and low voltage swing, Low Voltage Differential Signaling(LVDS) has been widely used for high speed data transmission with low power consumption. This paper proposes new LVDS I/O interface circuits for more than 1.3 Gb/s operation. The LVDS receiver proposed in this paper utilizes a sense amp for the pre-amp instead of a conventional differential pre-amp. The proposed LVDS allows more than 1.3 Gb/s transmission speed with significantly reduced driver output voltage. Also, in order to further improve the power consumption and noise performance, this paper introduces an inductance impedance matching technique which can eliminate the termination resistor. A new form of unfolded impedance matching method has been developed to accomplish the impedance matching for LVDS receivers with a sense amplifier as well as with a differential amplifier. The proposed LVDS I/O circuits have been extensively simulated using HSPICE based on 0.35um TSMC CMOS technology. The simulation results show improved power gain and transmission rate by ${\sim}12%$ and ${\sim}18%$, respectively.

Highly Sensitive Detection of Low-Abundance White Spot Syndrome Virus by a Pre-Amplification PCR Method

  • Pan, Xiaoming;Zhang, Yanfang;Sha, Xuejiao;Wang, Jing;Li, Jing;Dong, Ping;Liang, Xingguo
    • Journal of Microbiology and Biotechnology
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    • v.27 no.3
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    • pp.471-479
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    • 2017
  • White spot syndrome virus (WSSV) is a major threat to the shrimp farming industry and so far there is no effective therapy for it, and thus early diagnostic of WSSV is of great importance. However, at the early stage of infection, the extremely low-abundance of WSSV DNA challenges the detection sensitivity and accuracy of PCR. To effectively detect low-abundance WSSV, here we developed a pre-amplification PCR (pre-amp PCR) method to amplify trace amounts of WSSV DNA from massive background genomic DNA. Combining with normal specific PCR, 10 copies of target WSSV genes were detected from ${\sim}10^{10}$ magnitude of backgrounds. In particular, multiple target genes were able to be balanced amplified with similar efficiency due to the usage of the universal primer. The efficiency of the pre-amp PCR was validated by nested-PCR and quantitative PCR, and pre-amp PCR showed higher efficiency than nested-PCR when multiple targets were detected. The developed method is particularly suitable for the super early diagnosis of WSSV, and has potential to be applied in other low-abundance sample detection cases.

Characteristic as a Resonance Frequency of $SF_6$ Gas (SF6 가스중의 공진주파수에 따른 신호특성)

  • Lee, Y.H.;Lee, H.D.;Park, J.N.;Shin, Y.S.;Park, J.S.;Seo, J.M.
    • Proceedings of the KIEE Conference
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    • 2003.07c
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    • pp.1867-1869
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    • 2003
  • In this paper, chamber(Circuit breaker compartment of C-GIS) made of stainless steel with 4 mm width is used. Artificial defect was made on enclosure or HV conductor of chamber and $SF_6$ gas was injected into it according to pressure. In this experiment, Acoustic emission sensors of different types was used to compare sensitivity to detect acoustic signal occurred by Partial discharge(PD) of according to types and resonance frequency in $SF_6$ gas atmosphere. Sensors used in tests was R6I, R15I and 2/4/6 Pre-Amplifier connected with R6IU without pre. amp. In case of R6IU, gain was adjusted with 40 dB like other sensors and operated by differential mode. Post amplifier(post. amp) and band pass filter(BPF) were developed Gain of post. amp. is 60 dB and BPF has band width of $50{\sim}300$ kHz. Also, envelope circuit developed reduces frequency of AE sensor. As a result, in $SF_6$ atmosphere, R6IU and R6I had resonance frequency of 60 Hz was better than R15I. Also, R6IU was better than R6I because of type property of pre.amp. had differential mode.

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Improving the meiotic competence of small antral follicle-derived porcine oocytes by using dibutyryl-cAMP and melatonin

  • Jakree Jitjumnong;Pin-Chi Tang
    • Animal Bioscience
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    • v.37 no.6
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    • pp.1007-1020
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    • 2024
  • Objective: We increased the nuclear maturation rate of antral follicle derived oocytes by using a pre-in vitro maturation (IVM) culture system and improved the developmental potential of these porcine pathenotes by supplementing with melatonin. Furthermore, we investigated the expression patterns of genes involved in cumulus expansion (HAS2, PTGS2, TNFAIP6, and PTX3) derived from small and medium antral follicles before and after oocyte maturation. Methods: Only the cumulus oocyte-complexes (COCs) derived from small antral follicles were induced with [Pre-SF(+)hCG] or without [Pre-SF(-)hCG] the addition of human chorionic gonadotropin (hCG) during the last 7 h of the pre-IVM period before undergoing the regular culture system. The mature oocytes were investigated on embryonic development after parthenogenetic activation (PA). Melatonin (10-7 M) was supplemented during in vitro culture (IVC) to improve the developmental potential of these porcine pathenotes. Results: A pre-IVM culture system with hCG added during the last 7 h of the pre-IVM period [Pre-SF(+)hCG] effectively supported small antral follicle-derived oocytes and increased their nuclear maturation rate. The oocytes derived from medium antral follicles exhibited the highest nuclear maturation rate in a regular culture system. Compared with oocytes cultured in a regular culture system, those cultured in the pre-IVM culture system exhibited considerable overexpression of HAS2, PTGS2, and TNFAIP6. Porcine embryos treated with melatonin during IVC exhibited markedly improved quality and developmental competence after PA. Notably, melatonin supplementation during the IVM period can reduce and increase the levels of intracellular reactive oxygen species (ROS) and glutathione (GSH), respectively. Conclusion: Our findings indicate that the Pre-SF(+)hCG culture system increases the nuclear maturation rate of small antral follicle-derived oocytes and the expression of genes involved in cumulus expansion. Melatonin supplementation during IVC may improve the quality and increase the blastocyst formation rate of porcine embryos. In addition, it can reduce and increase the levels of ROS and GSH, respectively, in mature oocytes, thus affecting subsequent embryos.

Activation of CREB by PKA Promotes the Chondrogeneic Differentiation of Chick Limb Bud Mesenchymal Cells

  • Kim, Kook-Hee;Lee, Young-Sup
    • Animal cells and systems
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    • v.13 no.3
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    • pp.289-295
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    • 2009
  • Cyclic AMP-mediated signaling pathways regulate a number of cellular functions. In this study, we examined the regulatory role of cAMP signaling pathways in chondrogenesis of chick limb bud mesenchymal cells in vitro. Forskolin, which increases cellular cAMP levels by the activation of adenylate cyclase, enhanced chondrogenic differentiation. Inhibition of PKA with specific inhibitors (H89 or KT5720) blocked pre-cartilage condensation stage, indicating that chondrogenesis is regulated by the increase in cellular cAMP level and subsequent activation of PKA. Downstream signaling pathway of PKA leading to gene expression was investigated by examination of several nuclear transcription factors. Forskolin treatment increased transcription level for a cartilage-specific marker gene Sox9. However, inhibition of PKA with H89 led to restore expression of Sox9, indicating PKA activity was required to regulate the expression of Sox9 in chondrogenesis. In addition, CREB was highly phosphorylated at early stage of mesenchyme culture, and followed by progressive dephosphorylation. CBP and ATF, another CRE related proteins were transiently expressed at the early stage of chondrogenesis with a pattern similar to CREB phosphorylation. Electrophoretic mobility shift assays confirmed that the binding activity of CREB to the CRE is closely correlated to the phosphorylation pattern of CREB. Therefore, cAMP-mediated signal transduction to nuclear events for the induction of genes appeared to be required at the early stage of chick limb bud chondrogenesis.

The Effects of 3-Isobutyl-1-methylxanthine (IBMX) on Nuclear and Cytoplasmic Maturation of Porcine Oocytes In Vitro

  • Kwak, Seong-Sung;Jang, Seung-Hoon;Jeong, Se-Heon;Jeon, Yubyeol;Biswas, Dibyendu;Hyun, Sang-Hwan
    • Journal of Embryo Transfer
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    • v.27 no.3
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    • pp.163-169
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    • 2012
  • The 3-isobutyl-1-methylxanthine (IBMX) is non-selective phosphodiesterase and is able to prevent resumption of meiosis by maintaining elevated cyclic AMP (cAMP) concentrations in the oocyte. The present study was conducted to analyze: (1) nuclear maturation (examined by the Hoechst staining), (2) whether cytoplasmic maturation (examined by the intracellular glutathione (GSH) concentration) of porcine oocytes is improved during meiotic arrest after prematuration (22 h) with IBMX. Before in vitro maturation (IVM), oocytes were treated with 1 mM IBMX for 22 h. After 22 h of pre-maturation, the higher rate of IBMX treated group oocytes were arrested at the germinal vesicle (GV) stage (42.3%) than control IVM oocytes (10.1%). It appears that the effect of IBMX on the resumption of meiosis has shown clearly. In the end of IVM, the reversibility of the IBMX effect on the nuclear maturation has been corroborated in this study by the high proportions of MII stage oocytes (72.5%) reached after 44 h of IVM following the 22 h of inhibition. However, intracellular GSH concentrations were lower in the oocytes treated with IBMX than the control oocytes (6.78 and 12.94 pmol/oocyte, respectively). These results demonstrate that cytoplasmic maturation in porcine oocytes pre-treated with IBMX for 22 h did not equal that of control oocytes in the current IVM system. These results indicate that pre-maturation with IBMX for 22 h may not be beneficial in porcine IVM system.

The Effect of Articulation Modification Program for Production /ㅅ/ of Hearing Disordered Students (조음조절 프로그램에 의한 청각장애학생의 /ㅅ/산출 개선)

  • Son, Jung-Min;Seok, Dong-Il;Park, Sang-Hee
    • Speech Sciences
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    • v.7 no.3
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    • pp.229-247
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    • 2000
  • The purpose of this study is to examine the improvement effect of articulation of the fricative consonant /s/(ㅅ), when applying the Articulation Modification Program (AMP) to hearing disordered students. Three hearing disordered students were selected for this study by the consonant picture and Korean Articulation Sentence tests. They have no defect in their emotion, behavior and sight. This program applied the multiple baseline design across subjects to examine the improvement effect of articulation of fricative consonant /s/(ㅅ). Also I designed th$\sim$s program with an advanced pre-test and post-test in order to research the improvement difference of articulation by articulation position and language unit. It was executed 32 sessions over three months, four sessions a week, one session taking forty minutes. The results according to the study subject are as follow; First, as result of AMP, articulation of all students was developed after applying this program. Second, as result of AMP, articulation in fore-word was one-hundred percent efficiency and articulation in middle-word was 97.7% efficiency by position. Third, as result of AMP, articulation in the word reached 98.7% efficiency, in phrase 97.7% efficiency, in the sentence 97.7% efficiency and in the story was 98.3% efficiency by language unit.

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P2 Receptor-mediated Inhibition of Vasopressin-stimulated Fluid Transport and cAMP Responses in AQP2-transfected MDCK Cells

  • Kim, Yang-Hoo;Choi, Young-Jin;Bae, Hae-Rahn;Woo, Jae-Suk
    • The Korean Journal of Physiology and Pharmacology
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    • v.13 no.1
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    • pp.9-14
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    • 2009
  • We cultured canine kidney(MDCK) cells stably expressing aquaporin-2(AQP2) on collagen-coated permeable membrane filters and examined the effect of extracellular ATP on arginine vasopressin(AVP)-stimulated fluid transport and cAMP production. Exposure of cell monolayers to basolateral AVP resulted in stimulation of apical to basolateral net fluid transport driven by osmotic gradient which was formed by addition of 500 mM mannitol to basolateral bathing solution. Pre-exposure of the basolateral surface of cell monolayers to ATP(100 ${\mu}M$) for 30 min significantly inhibited the AVP-stimulated net fluid transport. In these cells, AVP-stimulated cAMP production was suppressed as well. Profile of the effects of different nucleotides suggested that the $P2Y_2$ receptor is involved in the action of ATP. ATP inhibited the effect of isoproterenol as well, but not that of forskolin to stimulate cAMP production. The inhibitory effect of ATP on AVP-stimulated fluid movement was attenuated by a protein kinase C inhibitor, calphostin C or pertussis toxin. These results suggest that prolonged activation of the P2 receptors inhibits AVP-stimulated fluid transport and cAMP responses in AQP2 transfected MDCK cells. Depressed responsiveness of the adenylyl cyclase by PKC-mediated modification of the pertussis-toxin sensitive $G_i$ protein seems to be the underlyihng mechanism.

Temporal Pattern of cAMP Concentrations and α-Actin mRNA Expression in Skeletal Muscle of Cimaterol-Fed Rats

  • Kim, Y.S.;Duguies, M.V.;Kim, Y.H.;Vincent, D.L.
    • Asian-Australasian Journal of Animal Sciences
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    • v.10 no.5
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    • pp.528-533
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    • 1997
  • Twenty four female Sprague-Dawley rats weighing about 190 g were used to examine changes in muscle cAMP concentrations and steady-state levels of skeletal muscle ${\alpha}$-actin mRNA during chronic administration of cimaterol, a ${\beta}$-adrenergic agonist. Cimaterol was mixed in a powdered rat diet at 10 mg/kg diet. At 3 and 21 days after the start of treatment, skeletal muscle and heart samples were collected for the measurement of cAMP concentrations and skeletal muscle ${\alpha}$-actin mRNA levels. Cimaterol increased (p < 0.01) body weight gain gradually during the first seven days of the trial period, but not thereafter. Most skeletal muscle weights and the ratio of muscle weight to body weight were increased (p < 0.05) by cimaterol treatment both at 3 and 21 days. Heart weight was also increased (p < 0.05) by cimaterol treatment at 3 and 21 days, but the ratio of heart weight to body weight was increased (p < 0.05) only at 3 day. Cimaterol decreased (p < 0.05) cAMP concentration of gastrocnemius muscle at both 3 and 21 days after treatment. However, cimaterol tended (p = 0.07) to increase cAMP concentration at 3 days in the heart. Cimaterol tended (p = 0.08) to increase the steady-state level of ${\alpha}$-actin mRNA by 60% in gastrocnemius muscle at 3 days but had no effect at 21 days. The results indicate that the pattern of hypertrophic response to chronic dietary administration of cimaterol is different between cardiac and skeletal muscle. In skeletal muscles it appears that the hypertrophy induced by cimaterol is partly due to stimulated myofibrillar protein synthesis at a pre-translational level.