• Molecules and Cells
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• 제44권2호
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• pp.88-100
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• 2021

Anoctamin 6/TMEM16F (ANO6) is a dual-function protein with Ca2+-activated ion channel and Ca2+-activated phospholipid scramblase activities, requiring a high intracellular Ca2+ concentration (e.g., half-maximal effective Ca2+ concentration [EC50] of [Ca2+]i > 10 μM), and strong and sustained depolarization above 0 mV. Structural comparison with Anoctamin 1/TMEM16A (ANO1), a canonical Ca2+-activated chloride channel exhibiting higher Ca2+ sensitivity (EC50 of 1 μM) than ANO6, suggested that a homologous Ca2+-transferring site in the N-terminal domain (Nt) might be responsible for the differential Ca2+ sensitivity and kinetics of activation between ANO6 and ANO1. To elucidate the role of the putative Ca2+-transferring reservoir in the Nt (Nt-CaRes), we constructed an ANO6-1-6 chimera in which Nt-CaRes was replaced with the corresponding domain of ANO1. ANO6-1-6 showed higher sensitivity to Ca2+ than ANO6. However, neither the speed of activation nor the voltage-dependence differed between ANO6 and ANO6-1-6. Molecular dynamics simulation revealed a reduced Ca2+ interaction with Nt-CaRes in ANO6 than ANO6-1-6. Moreover, mutations on potentially Ca2+-interacting acidic amino acids in ANO6 Nt-CaRes resulted in reduced Ca2+ sensitivity, implying direct interactions of Ca2+ with these residues. Based on these results, we cautiously suggest that the net charge of Nt-CaRes is responsible for the difference in Ca2+ sensitivity between ANO1 and ANO6.

• 한국수정란이식학회지
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• 제22권4호
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• pp.265-269
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• 2007

This study was conducted to investigate the efficacy of vitrification procedure for the cryopreservation of porcine oocytes and the utilization of vitrified oocytes as recipient cytoplasts for somatic cell nuclear transfer (NT), and observed that porcine oocytes are evaluated by pronuclear formation, and parthenogenetic development. Single fetal donor cells were deposited into the perivitelline space of vitrified enucleation oocytes, followed by electrical fusion and activation. NT embryos were cultured in NCSU-23 medium supplemented with 5% FBS, at $38.5^{\circ}C$ in 5% $CO_2$ and air. 1. When the developmental rates of the oocytes after being culture for $0{\sim}10$ hours vitrified with EDS and ETS were 42.0%, 38.0%, respectively. This results were lower than the control group(62.2%). 2. When the developmental rates of the oocytes after being culture for $0{\sim}10$ hours vitrified-thawed with sucrose and glucose, 5% PVP, NCSU-23 supplemented with 10% FBS were 33.3%, 25.9%, respectively. This results were lower than the control group(55.6%). 3. The fusion and development to the blastocyst stage between the NT embryos constructed with the vitrified and non-vitrified oocytes were significant differences. Developmental rate of oocytes and NT embryos constructed with the vitrified or non-vitrified oocytes were $13.0{\pm}2.4%\;and\;23.2{\pm}2.4%$, respectively.

• Reproductive and Developmental Biology
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• 제29권4호
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• pp.229-234
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• 2005

Methods for activation of reconstructed oocytes were examined for the production of nuclear transfer (NT) rat embryos using fetal neural stem cells as donor. Neural stem cells were isolated from Day 14.5 rat fetuses, and the oocytes for recipient cytoplasm were recovered from 4-week old Sprague Dawley rats. After enucleation and nuclear injection, the reconstructed oocytes were immediately exposed to activation medium consisting of 10 mM $SrCl_2$ for 4 h (immediate activation after injection; IAI), or cultured in vitro for $2\~3$ h before activation treatment (injection before activation; IBA). Pre-activated oocytes were also used for NT to test reprogramming potential of artificially activated oocytes. The oocytes were grouped as IIA (immediate injection after activation) and ABI (activation $2\~3$ h before injection). Following NT, the oocytes were cultured in vitro. Development of the NT embryos was monitored at 44 and 119 h after activation. The embryos in groups IAI, mA, and IIA were cleaved to the 2-cell stage at the rates of $36.6\%\;(15/41),\;39.5\%\;(17/43)\;and\;46.3\%$ (25/54), respectively. However, in the ABI group, only one embryo ($1.8\%$, 1/55) was cleaved after activation. After in vitro culture, two NT embryos from IAI group had developed to the morula stage $(4.9\%\cdot2/41)$. However, no morula or blastocyst was obtained in the other groups. These results suggest that immediate activation after injection (IAI) method may be used for the production of rat somatic cell NT embryos.

• Journal of Plant Biology
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• 제35권2호
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• pp.135-142
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• 1992

발아시킨 물오리나무(Alnus hirsuta)의 하배축 부위를 절단하여 얻은 떡잎이 포함된 상부를 6% glucose, MS(Murashige and Skoog, 1962) 비타민 혼합체, PVP(polyvinylpyrrolidone) 및 $0-50\;\mu\textrm{M}$이 BAP(6-benzylaminopurine)를 첨가한 NT(Nagata and Takebe, 1971) 무기염배지에서 배양한 결과, $5\;\mu\textrm{M}$이 BAP가 shoot 증식에 가장 적합한 것으로 나타났다. 따라서 이 배지에서 shoot 증식을 계속하여 실험에 사용할 200여개의 shoot를 확립하였다. Shoot 증식에 미치는 무기염류와 탄소원의 영향을 조사한 결과 탄소원에 무관하게 NT 무기염이 MS 무기염에 비해 3-12배의 증식을 나타냈으나 NT 무기염배지에서 탄소원은 특별한 차이를 보여주지 않았다. 발근에 미치는 NT 무기염, sucrose, IBA의 농도에 따른 영향을 조사한 결과 1/4-1/2 농도의 NT 무기염, 3%의 sucrose, $1-8\;\mu\textrm{M}$의 IBA에서 100% 가까운 발근율을 보였으며 뿌리발달도 양호하였다. 발근된 개체를 토양에 이식하고 습도를 점진적으로 낮추면서 적응시키면 거의 100%의 생존율을 나타내었다.

• The Korean Journal of Pain
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• 제18권1호
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• pp.29-33
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• 2005

Background: Chronic headache (CH) constitutes a significant public health problem, impacting on both the individual sufferer and society. Patients with CH, unresponsive to drug therapy or nerve block, suffer considerable disability due to the frequency and severity of attacks; therefore, they should be considered for novel therapy. Botulinum toxin type A (BoNT-A) has shown significant promise in the management of CH. In this paper, we review recent evidence on the efficacy of BoNT-A, and also report our experience with this treatment in CH patients. Methods: BoNT-A was used to treat 69 CH patients, including 47 in a chronic migraine group and 22 in a non-migraine CH group, who showed therapy-resistance to palliative drug or nerve block. We investigated the demography, dosage and site of BoNT-A injection, and used a visual analogue scale (VAS) for pain and the degree of satisfaction. The data were analyzed using t-tests and a Friedman repeated measures analysis of variance on ranks. Results: Significant decreases in the VAS for pain were found in both the chronic migraine and non-migraine CH groups, from 2, 4 and 12 weeks and from 4 and 12 weeks, respectively, after BoNT-A administration (P < 0.05). The chronic migraine group showed significantly lower VAS scores for pain than the non-migraine CH group from 2, 4 and 12 weeks after the BoNT-A administration (P < 0.05). Twenty eight patients (59.2%) in the chronic migraine group and eight (36.4%) in the non-migraine CH were satisfied with the BoNT-A treatment. Conclusions: This clinical study revealed that the use of BoNT-A demonstrated efficacy for CH patients resistant to drug therapy or nerve block. Moreover, BoNT-A proved itself more effective in the chronic migraine than non-migraine CH group.

• Reproductive and Developmental Biology
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• 제31권2호
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• pp.127-131
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• 2007

Embryonic germ (EG) cells are undifferentiated stern cells isolated from cultured primordial germ cells (PGC). These cells share many characteristics with embryonic stem cells including morphology and pluripotency. Undifferentiated porcine EG cell lines demonstrating capacities of differentiation both in vitro and in vivo have been established. Since EG cells can be cultured indefinitely in an undifferentiated state, whereas somatic cells in primary culture are often unstable and have limited lifespan, EG cells may provide inexhaustible source of karyoplasts in nuclear transfer (NT). In this study the efficiencies of NT using porcine EG and fetal fibroblast cells were compared. Two different techniques were used to perform NT. With conventional NT procedure (Roslin method) involving fusion of donor cells with enucleated oocytes, the rates of development to the blastocyst stage in EG and somatic cell NT were 16.8% (59/351) and 14.5% (98/677), respectively. In piezo-driven microinjection (Honolulu method) of donor nuclei into enucleated oocytes, the rates of blastocyst formation in EG and somatic cell NT were 11.9% (15/126) and 9.4% (9/96), respectively. Regardless of NT methods used in this study, EG cell NT gave rise to comparable rate of blastocyst development to somatic cell NT. Overall, EG cells can be used as karyoplast donor in NT procedure, and embryos can be produced by EG cell NT that may be used as an alternative to conventional somatic cell NT.

• 대한후두음성언어의학회지
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• 제30권2호
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• pp.77-81
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• 2019

Dysphagia may result from dysfunction of any of the components involved in the complex neuromuscular interaction of swallowing. Hyperfunction of any of the muscles involved in swallowing is a frequent cause of dysphagia. The cricopharyngeus muscle (CPM) is a key component of the upper esophageal sphincter. Cricopharyngeus muscle dysfunction (CPD) refers to the muscle's failure to appropriately and completely relax or expand during deglutition. A variety of disease processes may cause CPD and accurate diagnosis is paramount for appropriate treatment. In appropriately selected patients, intervention at the CPM may yield significant improvement in dysphagia. Interventions include nonsurgical, pharyngoesophageal segment dilatation, botulinum toxin (BoNT) injection, and criccopharyngeal myotomy. Injections of BoNT in patients with CPD have been reported to result in marked relief of dysphagia. Different techniques for instilling BoNT into the CPM have been described. Awake, in-office CPM BoNT injection with electromyography and/or fluoroscopic or ultrasound guidance is performed transcervically or via flexible endoscopy. Operative CPM BoNT injection involves rigid laryngoscopy and esophagoscopy with direct visualization of the CPM. BoNT should be prepared in low-volume, high-concentration dilutions to minimize the potential for undesired diffusion of the toxin. The effects of BoNT occur within weeks of injection and typically last up to 5 or 6 months.

• Reproductive and Developmental Biology
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• 제35권4호
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• pp.475-478
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• 2011

In the present study, the effect of cysteine and NT or bisphenol A(BP) on in vitro aturation(IVM) of porcine oocytes were examined. COCs was cultured in NCSU-23 medium supplement with 10% FCS which had previously been covered with mineral oil and equilibrated in a humidified atmosphere of 5% $CO_2$ and 95% air at $38^{\circ}C$. The IVM rate of oocytes cultured for 48 hrs in NCSU-23 medium supplement with 0.5~10.0 mM cysteine were $34.0{\pm}3.2%$, $36.0{\pm}3.5%$, $48.0{\pm}3.8%$, $22.0{\pm}3.2%$, respectively. The IVM rate of oocytes cultured in NCSU-23 medium supplement with 0.5~5.0mM NT for 48 hrs were $24.0{\pm}4.2%$, $18.0{\pm}4.9%$, $8.0{\pm}2.2%$, respectively. NT affects oocyte in vitro maturation rate in a dose-dependent. This result were significantly lower than the control group. The IVM rate of oocytes cultured for 48 hrs in NCSU-23 medium supplement with 1.0 mM NT+5.0 mM cysteine($38.0{\pm}4.3%$) were significantly higher than that of NT treatment. The IVM rate of oocytes cultured in NCSU-23 medium supplement with 0.05~5.0 mM BP for 48 hrs were $20.0{\pm}4.7%$, $10.0{\pm}5.3%$, $6.0{\pm}3.2%$, respectively. The IVM rate of oocytes cultured in NCSU-23 medium supplement with BP was significantly lower cultured non supplement of BP ($44.0{\pm}3.5%$). BP affects porcine oocyte maturation rate in a dose-dependent manner. The IVM rate of oocytes cultured for 48 hrs in NCSU-23 medium supplement with 1.0 mM BP+5.0 mM cycteine ($32.0{\pm}3.2%$) were increased than that of BP treatment.

• 한국신재생에너지학회:학술대회논문집
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• 한국신재생에너지학회 2011년도 추계학술대회 초록집
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• pp.100-100
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• 2011

Nanomaterial architecture with highly ordered, vertically oriented $TiO_2$ nanotube arrays shows a good promise for diverse technological applications. As inspired from the literature reports that Nickel modification can improve the photocatalytic activity of $TiO_2$, it was planned to coat Ni into the $TiO_2$ matrix. In this study, first $TiO_2$ nanotubes(TiNTs) were prepared by anodization (60V,3min) in HF-free aqueous electrolyte on ultrasonically cleaned polished titanium sheet substrates ($1{\times}7cm^2$). The typical thickness of the sintered TiNT ($500^{\circ}C$for10min) was ~1 micronas confirmed from the FESEM study. In the next part, as-anodized and sintered TiNT/Ti photoanodes were used to coat Ni by AC electrodeposition from aqueous 0.1M nickel sulphate solution. During AC electrodeposition, conditions such as 1V DC offset voltage, 9V amplitude (peak-to-peak) and 750 Hz frequency were fixed constant and the deposition time was varied as 0.5 min, 1 min, 2 min and 10 min. The photoelectrochemical performance of pristine and Ni modified TiNT/Ti photoanodes was measured in 1N NaOH electrolyte under 1 SUN illumination in the potential range of -1V and 1.2V versus Ag/AgCl reference electrode. The photocurrent performance of TiNT/Ti photoanode decreased upon Ni modification and the results were confirmed after repeated experiments. This suggests us that Ni modification inhibits the photoelectrochemical performance of $TiO_2$ nanotubes.

• Journal of Genetic Medicine
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• 제11권2호
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• pp.56-62
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• 2014

Purpose: To assess the outcomes of increased fetal nuchal translucency (NT), to aid in prenatal counseling and management in our practice. Materials and Methods: We retrospectively reviewed the medical records of patients who underwent first trimester fetal karyotyping using chorionic villi sampling (CVS) and second trimester level II sonography for a fetal NT thickness ${\geq}3.0mm$ between 11 weeks and 13 weeks 6 days' gestation, at Gyeongsang National University Hospital. Pediatric medical records and a telephone interview were used to follow-up live-born children. Exclusion criteria included incomplete data and CVS for other indications. Results: Seventy cases met the inclusion criteria (median NT thickness, 4.7 mm; range, 3.0-16.1 mm). Twenty-nine cases (41.4%) were aneuploid. The prevalence of chromosomal defects increased with NT thickness: NT 3.0-3.4 mm, 16.7%; NT 3.5-4.4 mm, 27.3%; NT 4.5-5.4 mm, 66.7%; NT 5.5-6.4 mm, 37.5%; NT ${\geq}6.5mm$, 62.5%. The most common karyotype abnormality was trisomy 18 (n=12), followed by trisomy 21 (n=9). In chromosomally normal fetuses (n=41), fetal death occurred in 2 cases (4.9%), and structural malformations were found in 11 cases (26.8%). In chromosomally and anatomically normal fetuses (n=28), one child had neurodevelopmental delay (3.6%). Twenty-eight infants who had a prenatal increased NT were alive and well at follow-up (40%). Conclusion: Outcomes of increased fetal NT might help inform prenatal counseling and management. The high prevalence of chromosomal defects associated with increased fetal NT implies that CVS should be performed in the first trimester, particularly considering the stress associated with an uncertain diagnosis.