• Title/Summary/Keyword: Pipette

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Granulometric Analysis of Sediment: Preliminary Results for Inter-lab Calibration (퇴적물 입도분석 방식 비교: 예비결과)

  • Choi, J.Y.;Choi, J.H.;Kim, S.Y.
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.4 no.3
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    • pp.246-253
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    • 1999
  • Grain size distributions of sediment samples were analyzed by the different methods, and the results were compared. Reported data of the bottom sediments from the Korean seas show significant deviations among the institutes, and the inter-lab comparison and calibration procedures are considered to be necessary. Grain size data by different methods show different results. Laser diffraction method provides good precision in replicate analysis, but underestimates the amounts of finer (smaller than 2-3 ${\mu}m$) fraction. Data from particle settling method, on the other hand, represent significant errors in the coarse silt (5-6 ${\phi}$) fraction, and slightly overestimate the clay fraction. In the sieve and pipette methods, best results were taken in 0.5 ${\phi}$ interval of analysis. Especially in the coarse silt fraction, pipette analysis is suggested to be made with 0.5 ${\phi}$ interval. During the dry sieving procedure in 1 ${\phi}$ interval, serious errors occur when large amounts of sample materials were concentrated in a particular sieve.

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Ionic Dependence and Modulatory Factors of the Background Current Activated by Isoprenaline in Rabbit Ventricular Cells

  • Leem, Chae-Hun;Lee, Suk-Ho;So, In-Suk;Ho, Won-Kyung;Earm, Yung-E
    • The Korean Journal of Physiology
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    • v.26 no.1
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    • pp.15-25
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    • 1992
  • In order to elucidate the properties of the background current whole cell patch clamp studies were performed in rabbit ventricular cells. Ramp pulses of ${\pm}80\;mV$ from holding potential of 40 mV(or 20 mV) at the speed of 0.8 V/sec were given every 30 sec(or 10 sec) and current-voltage diagrams(I-V curve) were obtained. For the activation of the background current isoprenaline, adenosine 3',5'-cyclic monophosphate(dBcAMP), guanosine 3',5'-cyclic monophosphate(cGMP), and $N^6$-2'-o-dibutyryladenosine 3',5'-cyclic monophosphate(dBcAMP) were applied after all known current systems were blocked with 2mM Ba, 1 mM Cd ,5 mM Ni, 10 ${\mu}M$ diltiazem, 10 ${\mu}m$ ouabain, and 20 mM tetraethylammonium(TEA). The conductance of background current in control was $0.65{\pm}0.69$ nS at 0 mV, its I-V curves was almost linear and reversed near 50 mV. When there was no taurine in pipette solution, isoprenaline hardly activated the background current but when taurine existed in pipette solution, isoprenaline activated the larger background current. Cyclic AMP or cyclic GMP alone had little effect on the activation of the background current, while cGMP potentiated cGMP effect. When the background current was activated with cGMP and cAMP, isoprenaline could not further increased the background current. The background current activated by isoprenaline depended on extracellular $Cl^-$ concentration and its reversal potential was shifted according to chloride equilibrium potential. The change of extracellular $Na+$ concentration had little effect on reversal potential of the background current activated by isoprenaline.

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Voltage-dependent $Ca^{2+}$ Current Identified in Freshly Isolated Interstitial Cells of Cajal (ICC) of Guinea-pig Stomach

  • Kim, Young-Chul;Suzuki, Hikaru;Xu, Wen-Xie;Hashitani, Hikaru;Choi, Woong;Yun, Hyo-Yung;Park, Seon-Mee;Youn, Sei-Jin;Lee, Sang-Jeon;Lee, Sang-Jin
    • The Korean Journal of Physiology and Pharmacology
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    • v.12 no.6
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    • pp.323-330
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    • 2008
  • The properties of voltage dependent $Ca^{2+}$ current (VDCC) were investigated in interstitial cells of Cajal (ICC) distributed in the myenteric layer (ICC-MY) of guinea-pig antrum. In tissue, ICC-MY showed c-Kit positive reactions and produced driving potentials with the amplitude and frequency of about 62 mV and 2 times $min^{-1}$ respectively, in the presence of $1{\mu}M$ nifedipine. Single ICC-MY isolated by enzyme treatment also showed c-Kit immunohistochemical reactivity. These cells were also identified by generation of spontaneous inward current under $K^+$ -rich pipette solution. The voltage clamp experiments revealed the amplitude of - 329 pA inward current at irregular frequency. With $Cs^+$-rich pipette solution at $V_h=-80\;mV$, ICC-MY produced voltage-dependent inward currents (VDIC), and nifedipine ($1{\mu}M$) blocked VDIC. Therefore, we successfully isolated c-Kit positive single ICC from guinea-pig stomach, and found that ICC-MY potently produced dihydropiridine sensitive L-type voltage-dependent $Ca^{2+}$ currents ($VDCC_L$).

Effects of Noradrenaline on the Membrane Potential of Prostatic Neuroendocrine Cells of Rat

  • Kim, Jun-Hee;Shin, Sun-Young;Uhm, Dae-Yong;Kim, Sung-Joon
    • The Korean Journal of Physiology and Pharmacology
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    • v.7 no.1
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    • pp.47-52
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    • 2003
  • The prostate gland contains numerous neuroendocrine cells that are believed to influence the function of the prostate gland. Our recent study demonstrated the expression of both ${\alpha}1$- and ${\alpha}2$-ARs, signaling the release of stored $Ca^{2+}$ and the inhibition of N-type $Ca^{2+}$ channels, respectively, in rat prostate neuroendocrine cells (RPNECs). In this study, the effects of NA on the resting membrane potential (RMP) of RPNECs were investigated using a whole-cell patch clamp method. Fresh RPNECs were dissociated from the ventral lobe of rat prostate and identified from its characteristic shape; round or oval shape with dark cytoplasm. Under zero-current clamp conditions with KCl pipette solution, the resting membrane potential (RMP) of RPNECs was between -35 mV and -85 mV. In those RPNECs with relatively hyperpolarized RMP (<-60 mV), the application of noradrenaline (NA, $1{\mu}M$) depolarized the membrane to around -40 mV. In contrast, the RPNECs with relatively depolarized RMP (>-45 mV) showed a transient hyperpolarization and subsequent fluctuation at around -40 mV on application of NA. Under voltage clamp conditions (holding voltage, -40 mV) with CsCl pipette solution, NA evoked a slight inward current (<-20 pA). NA induced a sharp increase of cytosolic $Ca^{2+}$ concentration ($[Ca^{2+}]_c$), measured by the fura-2 fluorescence, and the voltage clamp study showed the presence of charybdotoxin-sensitive $Ca^{2+}$-activated $K^+$ currents. In summary, adrenergic stimulation induced either depolarization or hyperpolarization of RPNECs, depending on the initial level of RMP. The inward current evoked by NA and the $Ca^{2+}$-activated $K^+$ current might partly explain the depolarization and hyperpolarization, respectively.

Fabrication of a Micro-thermoelectric Probe (마이크로 프로브 기반 열전 센서 제작 기술)

  • Chang, Won-Seok;Choi, Tae-Youl
    • Transactions of the Korean Society of Mechanical Engineers B
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    • v.35 no.11
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    • pp.1133-1137
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    • 2011
  • A novel technique for the fabrication of a glass micropipette-based thermal sensor was developed utilizing inexpensive thermocouple materials. Thermal fluctuation with a resolution of ${\pm}0.002$ K was measured using the fabricated thermal probe. The sensors comprise unleaded low-melting point solder alloy (Sn) as a core metal inside a borosilicate glass pipette coated with a thin film of Ni, creating a thermocouple junction at the tip. The sensor was calibrated using a thermally insulated calibration chamber, the temperature of which can be controlled with a precision of ${\pm}0.1$ K and the thermoelectric power (Seebeck coefficient) of the sensor was recorded from 8.46 to $8.86{\mu}V$/K. The sensor we have produced is both cost-effective and reliable for thermal conductivity measurements of micro-electromechanical systems (MEMS) and biological temperature sensing at the micron level.

Glycine- and GABA-mimetic Actions of Shilajit on the Substantia Gelatinosa Neurons of the Trigeminal Subnucleus Caudalis in Mice

  • Yin, Hua;Yang, Eun-Ju;Park, Soo-Joung;Han, Seong-Kyu
    • The Korean Journal of Physiology and Pharmacology
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    • v.15 no.5
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    • pp.285-289
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    • 2011
  • Shilajit, a medicine herb commonly used in Ayurveda, has been reported to contain at least 85 minerals in ionic form that act on a variety of chemical, biological, and physical stressors. The substantia gelatinosa (SG) neurons of the trigeminal subnucleus caudalis (Vc) are involved in orofacial nociceptive processing. Shilajit has been reported to be an injury and muscular pain reliever but there have been few functional studies of the effect of Shilajit on the SG neurons of the Vc. Therefore, whole cell and gramicidin-perfotrated patch clamp studies were performed to examine the action mechanism of Shilajit on the SG neurons of Vc from mouse brainstem slices. In the whole cell patch clamp mode, Shilajit induced short-lived and repeatable inward currents under the condition of a high chloride pipette solution on all the SG neurons tested. The Shilajit-induced inward currents were concentration dependent and maintained in the presence of tetrodotoxin (TTX), a voltage gated $Na^+$ channel blocker, CNQX, a non-NMDA glutamate receptor antagonist, and AP5, an NMDA receptor antagonist. The Shilajit-induced responses were partially suppressed by picrotoxin, a $GABA_A$ receptor antagonist, and totally blocked in the presence of strychnine, a glycine receptor antagonist, however not affected by mecamylamine hydrochloride (MCH), a nicotinic acetylcholine receptor antagonist. Under the potassium gluconate pipette solution at holding potential 0 mV, Shilajit induced repeatable outward current. These results show that Shilajit has inhibitory effects on the SG neurons of Vc through chloride ion channels by activation of the glycine receptor and $GABA_A$ receptor, indicating that Shilajit contains sedating ingredients for the central nervous system. These results also suggest that Shilajit may be a potential target for modulating orofacial pain processing.

Regulation of Transient Receptor Potential Melastatin 7 (TRPM7) Currents by Mitochondria

  • Kim, Byung Joo;Jeon, Ju-Hong;Kim, Seon Jeong;So, Insuk;Kim, Ki Whan
    • Molecules and Cells
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    • v.23 no.3
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    • pp.363-369
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    • 2007
  • Mitochondria play a central role in energy-generating processes and may be involved in the regulation of channels and receptors. Here we investigated TRPM7, an ion channel and functional kinase, and its regulation by mitochondria. Proton ionophores such as CCCP elicited a rapid decrease in outward TRPM7 whole-cell currents but a slight increase in inward currents with pipette solutions containing no MgATP. With pipette solutions containing 3 mM MgATP, however, CCCP increased both outward and inward TRPM7 currents. This effect was reproducible and fully reversible, and repeated application of CCCP yielded similar decreases in current amplitude. Oligomycin, an inhibitor of $F_1/F_O$-ATP synthase, inhibited outward whole-cell currents but did not affect inward currents. The respiratory chain complex I inhibitor, rotenone, and complex III inhibitor, antimycin A, were without effect as were kaempferol, an activator of the mitochondrial $Ca^{2+}$ uniporter, and ruthenium red, an inhibitor of the mitochondrial $Ca^{2+}$ uniporter. These results suggest that the inner membrane potential (as regulated by proton ionophores) and the $F_1/F_O$-ATP synthase of mitochondria are important in regulating TRPM7 channels.

A Study on Developing Reliable Standard Samples of Crystalline Silica (결정형 산화규소의 표준시료 확보방안에 대한 연구)

  • Eun-song Hwang;Eun-Ji Lee;Hae Dong Park
    • Journal of Korean Society of Occupational and Environmental Hygiene
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    • v.34 no.3
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    • pp.247-253
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    • 2024
  • Objectives: The purpose of this study is to establish a method for the production of standard samples of crystalline silica and to assess its feasibility for the future application in non-mandatory quality control. Methods: We dispersed crystalline silica in 2-propanol to prepare a 1 mg/mL dispersion solution. Between 50-200 uL of the dispersion solution was taken with a pipette, injected onto PVC filters, air-dried, and used as samples. The variation and homogeneity of the quartz concentrations were confirmed by FTIR analysis. Analytical proficiency testing was conducted across 30 voluntary participants. Results: The coefficient of variation was lower for samples prepared using supernatant (settled for 16 hours) compared to those made with dispersion solution. The contamination level after using the pipette tip one to five times was 8.13-9.89% of the solution's concentration. Homogeneity was confirmed within the concentration of 0.0223-0.0431 mg/mL, but it was not at the higher concentration of 0.0553 mg/mL. In the analytical proficiency testing, the coefficient of variation for each level ranged from 24.6% to 28.3%, with a compliance rate of 90.0% to 96.7%. Conclusions: This study was conducted to produce standard samples of crystalline silica. The homogeneity of the samples was confirmed at low concentrations. However, further investigations on the applicability of wider range of concentrations are needed.

The Significance of Acetylcholine Receptor Autoantibody Test (아세틸콜린 수용체 항체(Acetylcholine receptor autoantibody) 검사의 의의)

  • Yoo, Soh-Yeon;Lim, Soo-Yeon;Pack, Song-Ran;Seo, Mi-Hye;Moon, Hyung-Ho;You, Sun-Hee
    • The Korean Journal of Nuclear Medicine Technology
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    • v.15 no.1
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    • pp.113-116
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    • 2011
  • Purpose: Acetylcholine receptor antibodies cause acetylcholine receptor loss, which is responsible for failure of the neuromuscular junction in the acetylcholine receptor autoantibody. The disease characterized by muscle weakness and fatigue, myasthenia gravis(MG) occurs when the body inappropriately produces antibodies against acetylcholine receptors, and thus inhibits proper acetylcholine signal transmission. And this reason, the measurement of acetylcholine receptor antibodies can be of considerable value in disease diagnosis. Methods: From 2010. August to September, we tested orderd AchRAb 19 samples to get the results. 1. Pipette $5{\mu}{\ell}$ undiluted patient sera and kit control and add 125I AChR $50{\mu}{\ell}$ and incubate at R.T for 2 hours. 2. Pipette $50{\mu}{\ell}$ of anti-human IgG into each tube, and incubate at $2{\sim}8^{\circ}C$ for 2 hours. 3. Pipette $25{\mu}{\ell}$ precipitation enhancer into each tube and add 1mL washing solution into all tubes. 4. Centrifuge each tube for 20minutes at $2{\sim}8^{\circ}C$ at 1500g. 5. Aspirate or decant the supernatant. 6. Pipette 1 mL washing solution into all tubes and resuspend the pellet and repeat centrifugation. 7. Aspirate or decant the supernatant and count all tubes on a gamma counter. Results: Cut off value is 0.2 nmol/L and the results taken below 0.2 nmol/L are negative, the results above that identified as being positive values. We assayed the 19 patients samples and got 7 positive results. Of which, 6 patients were diagnosed as MG.(85.7%). Conclusions: Acetylcholine Receptor autoantibody test is intended for use by persons only for the quantitative determination of it in human serum. Even if measurement of the antibodies is not a routine test, it can be of considerable value in disease diagnosis.

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Establishment of An Efficient and Stable Transgene Expression System in Chicken Primordial Germ Cells

  • Yang, Ju-Hyun;Kim, Sung-Tae
    • Bulletin of the Korean Chemical Society
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    • v.33 no.5
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    • pp.1536-1540
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    • 2012
  • Chicken primordial germ cells (cPGCs) are founder germ cells in embryonic stage of development that eventually give rise to sperms or oocytes. Currently cPGCs are only known cells enabling germline transmission in chicken and their cultivation protocols were recently established. Although genome modifications of chickens are now theoretically possible using cPGCs, there are still several hurdles to overcome to practically use cPGCs as mediators for chicken transgenesis. First, efficiency of gene delivery into cPGCs remains low with current methods. Second, there aregene silencing mechanisms against the expression of foreign genes in cPGCs. In this study, we successfully increased the efficiency of gene delivery in cPGCs by taking advantage of the TTAA-specific $piggybac$ transposon system. Moreover, a pipette-type electroporator significantly enhanced transfection efficiency up to 5-fold compared withcuvette-type methods. Taken together, the technological advances in our study will provide practical benefits for the application to fulfill genetic modifications of chicken genome.