• Molecules and Cells
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• v.39 no.4
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• pp.322-329
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• 2016

Voltage-gated $Ca^{2+}$ ($Ca_V$) channels are dynamically modulated by Gprotein-coupled receptors (GPCR). The $M_1$ muscarinic receptor stimulation is known to enhance $Ca_V2.3$ channel gating through the activation of protein kinase C (PKC). Here, we found that $M_1$ receptors also inhibit $Ca_V2.3$ currents when the channels are fully activated by PKC. In whole-cell configuration, the application of phorbol 12-myristate 13-acetate (PMA), a PKC activator, potentiated $Ca_V2.3$ currents by ~two-fold. After the PMA-induced potentiation, stimulation of $M_1$ receptors decreased the $Ca_V2.3$ currents by $52{\pm}8%$. We examined whether the depletion of phosphatidylinositol 4,5-bisphosphate ($PI(4,5)P_2$) is responsible for the muscarinic suppression of $Ca_V2.3$ currents by using two methods: the Danio rerio voltage-sensing phosphatase (Dr-VSP) system and the rapamycin-induced translocatable pseudojanin (PJ) system. First, dephosphorylation of $PI(4,5)P_2$ to phosphatidylinositol 4-phosphate (PI(4)P) by Dr-VSP significantly suppressed $Ca_V2.3$ currents, by $53{\pm}3%$. Next, dephosphorylation of both PI(4)P and $PI(4,5)P_2$ to PI by PJ translocation further decreased the current by up to $66{\pm}3%$. The results suggest that $Ca_V2.3$ currents are modulated by the $M_1$ receptor in a dual mode-that is, potentiation through the activation of PKC and suppression by the depletion of membrane $PI(4,5)P_2$. Our results also suggest that there is rapid turnover between PI(4)P and $PI(4,5)P_2$ in the plasma membrane.

• Molecules and Cells
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• v.22 no.1
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• pp.97-103
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• 2006

Phosphoinositides are critical regulators of ion channel and transporter activity. There are multiple isomers of biologically active phosphoinositides in the plasma membrane and the different lipid species are non-randomly distributed. However, the mechanism by which cells impose selectivity and directionality on lipid movements and so generate a non-random lipid distribution remains unclear. In the present study we investigated which structural elements of phosphoinositides are responsible for their subcellular location and movement. We incubated phosphatidylinositol (PI), phosphatidylinositol 4-monophosphate (PI(4)P) and phosphatidylinositol 4,5-bisphosphate ($PI(4,5)P_2$) with short or long acyl chains in CHO and HEK cells. We show that phosphate number and acyl chain length determine cellular location and translocation movement. In CHO cells, $PI(4,5)P_2$ with a long acyl chain was released into the cytosol easily because of a low partition coefficient whereas long chain PI was released more slowly because of a high partition coefficient. In HEK cells, the cellular location and translocation movement of PI were similar to those of PI in CHO cells, whereas those of $PI(4,5)P_2$ were different; some mechanism restricted the translocation movement of $PI(4,5)P_2$, and this is in good agreement with the extremely low lateral diffusion of $PI(4,5)P_2$. In contrast to the dependence on the number of phosphates of the phospholipid head group of long acyl chain analogs, short acyl chain phospholipids easily undergo translocation movement regardless of cell type and number of phosphates in the lipid headgroup.

• 한국약용작물학회:학술대회논문집
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• 2011.09a
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• pp.31-45
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• 2011

The amyloid ${\beta}$-peptide ($A{\beta}$), which originates from the proteolytic cleavage of amyloid precursor protein (APP), plays a central role in the pathogenesis of Alzheimer's disease (AD). Mounting evidence indicates that different species of $A{\beta}$, such as $A{\beta}$ oligomers and fibrils, may contribute to AD pathogenesis via distinct mechanisms at different stages of the disease. Importantly, elevation and accumulation of soluble $A{\beta}$ oligomers closely correlate with cognitive decline and/or disease progression in animal models of AD. In agreement with these studies, oligomers of $A{\beta}$ have been shown to directly affect synaptic plasticity, a neuronal process that is known to be essential for memory formation. Our previous studies showed that $A{\beta}$ induces the breakdown of phosphatidylinositol 4,5-bisphosphate (PI(4,5)P2), a phospholipid that regulates key aspects of neuronal function. PI(4,5)P2 breakdown was found to be a key step toward synaptic and memory dysfunction in a mouse model of AD. To this end, we seek to identify small molecules that could elevate the levels of PI(4,5)P2 and subsequently block $A{\beta}$ oligomer-induced breakdown of PI(4,5)P2 and synaptic dysfunction.. We found that (20S)Rg3, an active triterpene glycoside from heat-processed ginseng, serves as an agonist for phosphatidylinositol 4-kinase IIalpha (PI4KIIalpha), which is a lipid kinase that mediates a rate-limiting step in PI(4,5)P2 synthesis. Consequently, (20S)Rg3 stimulates PI(4,5)P2 synthesis by directly stimulating the activity of PI4KIIalpha. Interestingly, treatment of a mouse model of AD with (20S)Rg3 leads to reversal of memory deficits. Our data suggest that the PI(4,5)P2-promoting effects of (20S)Rg3 may help mitigate the cognitive symptoms associated with AD.

• Korean Journal of Radiology
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• v.22 no.7
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• pp.1100-1109
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• 2021

Objective: To compare the diagnostic performance between Prostate Imaging-Reporting and Data System version 2.0 (PI-RADSv2.0) and version 2.1 (PI-RADSv2.1) for clinically significant prostate cancer (csPCa) in the peripheral zone (PZ). Materials and Methods: This retrospective study included 317 patients who underwent multiparametric magnetic resonance imaging and targeted biopsy for PZ lesions. Definition of csPCa was International Society of Urologic Pathology grade ≥ 2 cancer. Area under the curve (AUC), sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and accuracy for csPCa were analyzed by two readers. The cancer detection rate (CDR) for csPCa was investigated according to the PI-RADS categories. Results: AUC of PI-RADSv2.1 (0.856 and 0.858 for reader 1 and 2 respectively) was higher than that of PI-RADSv2.0 (0.795 and 0.747 for reader 1 and 2 respectively) (both p < 0.001). Sensitivity, specificity, PPV, NPV, and accuracy for PI-RADSv2.0 vs. PI-RADSv2.1 were 93.2% vs. 88.3% (p = 0.023), 52.8% vs. 76.6% (p < 0.001), 48.7% vs. 64.5% (p < 0.001), 94.2% vs. 93.2% (p = 0.504), and 65.9% vs. 80.4% (p < 0.001) for reader 1, and 96.1% vs. 92.2% (p = 0.046), 34.1% vs. 72.4% (p < 0.001), 41.3% vs. 61.7% (p < 0.001), 94.8% vs. 95.1% (p = 0.869), and 54.3% vs. 78.9% (p < 0.001) for reader 2, respectively. CDRs of PI-RADS categories 1-2, 3, 4, and 5 for PI-RADSv2.0 vs. PI-RADSv2.1 were 5.9% vs. 5.9%, 5.8% vs. 12.5%, 39.8% vs. 56.2%, and 88.9% vs. 88.9% for reader 1; and 4.5% vs. 4.1%, 6.1% vs. 11.1%, 32.5% vs. 53.4%, and 85.0% vs. 86.8% for reader 2, respectively. Conclusion: Our data demonstrated improved AUC, specificity, PPV, accuracy, and CDRs of category 3 or 4 of PI-RADSv2.1, but decreased sensitivity, compared with PI-RADSv2.0, for csPCa in PZ.

• BMB Reports
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• v.47 no.7
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• pp.361-368
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• 2014

Lipid components in biological membranes are essential for maintaining cellular function. Phosphoinositides, the phosphorylated derivatives of phosphatidylinositol (PI), regulate many critical cell processes involving membrane signaling, trafficking, and reorganization. Multiple metabolic pathways including phosphoinositide kinases and phosphatases and phospholipases tightly control spatio-temporal concentration of membrane phosphoinositides. Metabolizing enzymes responsible for PI 4,5-bisphosphate (PI(4,5)P2) production or degradation play a regulatory role in Toll-like receptor (TLR) signaling and trafficking. These enzymes include PI 4-phosphate 5-kinase, phosphatase and tensin homolog, PI 3-kinase, and phospholipase C. PI(4,5)P2 mediates the interaction with target cytosolic proteins to induce their membrane translocation, regulate vesicular trafficking, and serve as a precursor for other signaling lipids. TLR activation is important for the innate immune response and is implicated in diverse pathophysiological disorders. TLR signaling is controlled by specific interactions with distinct signaling and sorting adaptors. Importantly, TLR signaling machinery is differentially formed depending on a specific membrane compartment during signaling cascades. Although detailed mechanisms remain to be fully clarified, phosphoinositide metabolism is promising for a better understanding of such spatio-temporal regulation of TLR signaling and trafficking.

• BMB Reports
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• v.54 no.6
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• pp.311-316
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• 2021

Ethanol often causes critical health problems by altering the neuronal activities of the central and peripheral nerve systems. One of the cellular targets of ethanol is the plasma membrane proteins including ion channels and receptors. Recently, we reported that ethanol elevates membrane excitability in sympathetic neurons by inhibiting Kv7.2/7.3 channels in a cell type-specific manner. Even though our studies revealed that the inhibitory effects of ethanol on the Kv7.2/7.3 channel was diminished by the increase of plasma membrane phosphatidylinositol 4,5-bisphosphate (PI(4,5)P₂), the molecular mechanism of ethanol on Kv7.2/7.3 channel inhibition remains unclear. By investigating the kinetics of Kv7.2/7.3 current in high K⁺ solution, we found that ethanol inhibited Kv7.2/7.3 channels through a mechanism distinct from that of tetraethylammonium (TEA) which enters into the pore and blocks the gate of the channels. Using a non-stationary noise analysis (NSNA), we demonstrated that the inhibitory effect of ethanol is the result of reduction of open probability (P_O) of the Kv7.2/7.3 channel, but not of a single channel current (i) or channel number (N). Finally, ethanol selectively facilitated the kinetics of Kv7.2 current suppression by voltage-sensing phosphatase (VSP)-induced PI(4,5)P₂ depletion, while it slowed down Kv7.2 current recovery from the VSP-induced inhibition. Together our results suggest that ethanol regulates neuronal activity through the reduction of open probability and PI(4,5)P₂ sensitivity of Kv7.2/7.3 channels.

• Sleep Medicine and Psychophysiology
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• v.19 no.2
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• pp.68-76
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• 2012

Objectives: Sleep disorders cause changes of autonomic nervous system (ANS) which affect cardiovascular system. Primary insomnia (PI) makes acceleration of sympathetic nervous system (SNS) tone by sleep deficiency and arousal. Obstructive sleep apnea syndrome (OSAS) sets off SNS by frequent arousals and hypoxemias during sleep. We aimed to compare the changes of heart rate variability (HRV) indices induced by insomnia or sleep apnea to analyze for ANS how much to be affected by PI or OSAS. Methods: Total 315 subjects carried out nocturnal polysomnography (NPSG) were categorized into 4 groups - PI, mild, moderate and severe OSAS. Severity of OSAS was determined by apnea-hypopnea index (AHI). Then we selected 110 subjects considering age, sex and valance of each group's size [Group 1 : PI (mean age=$41.50{\pm}13.16$ yrs, AHI <5, n=20), Group 2 : mild OSAS (mean age=$43.67{\pm}12.11$ yrs, AHI 5-15, n=30), Group 3 : moderate OSAS (mean age $44.93{\pm}12.38$ yrs, AHI 16-30, n=30), Group 4 : severe OSAS (mean age=$45.87{\pm}12.44$ yrs, AHI >30, n=30)]. Comparison of HRV indices among the four groups was performed with ANCOVA (adjusted for age and body mass index) and Sidak post-hoc test. Results: We found statistically significant differences in HRV indices between severe OSAS group and the other groups (PI, mild OSAS and moderate OSAS). And there were no significant differences in HRV indices among PI, mild and moderate OSAS group. In HRV indices of PI and severe OSAS group showing the most prominent difference in the group comparisons, average RR interval were $991.1{\pm}27.1$ and $875.8{\pm}22.0$ ms (p=0.016), standard deviation of NN interval (SDNN) was $85.4{\pm}6.6$ and $112.8{\pm}5.4$ ms (p=0.022), SDNN index was $57.5{\pm}5.2$ and $87.6{\pm}4.2$ (p<0.001), total power was $11,893.5{\pm}1,359.9$ and $18,097.0{\pm}1,107.2ms^2$(p=0.008), very low frequency (VLF) was $7,534.8{\pm}1,120.1$ and $11,883.8{\pm}912.0ms^2$ (p=0.035), low frequency (LF) was $2,724.2{\pm}327.8$ and $4,351.6{\pm}266.9ms^2$(p=0.003). Conclusions: VLF and LF which were correlated with SNS tone showed more increased differences between severe OSAS group and PI group than other group comparisons. We could suggest that severe OSAS group was more influential to increased SNS activity than PI group.

• Korean Journal of Plant Tissue Culture
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• v.25 no.1
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• pp.45-50
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• 1998

Hypocotyl explants of flowering cabbage were precultured on MS medium without kanamycin and then cocultured with Agrobacterium tumefaciens LBA4404;;pGA875 harboring insect resistantce proteinase inhibitor II(PI-II) gene in MS liquid medium adjusted pH 5.5 for 72hr. These explants were transferred to MS medium containing 20 mg/L kanamycin, 500 mg/L carbenicillin, and 1 mg/L BA. The explants were subsequently subcultured every 2 weeks. After 4 weeks of subculture, kanamycin-resistant shoots were obtained from selection medium. Leaves of putative transformants survived on MS selection medium containing 30 mg/L kanamycin. Incoporation of the PI-II gene into flowering cabbage was confirmed by PCR analysis of genomic DNA. Southern blot analysis showed that ECL-labeled probe for PI-II gene was hybridized to the expected amplified genomic DNA fragment of about 500 by from transgenic flowering cabbage.

• Journal of Life Science
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• v.26 no.8
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• pp.875-880
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• 2016

The plasma membrane plays a crucial role in relaying signals from the outside environment to the inside of the cells. In eukaryotic cells, the inner leaflets of the plasma membrane are composed mostly of phospholipids, including phosphatidylethanolamine (PE), phosphatidylserine (PS), and phosphatidylinositides (PIs). In this study, we tried to analyze the morphological changes induced by EGFP-fused membrane binding proteins, which are targeted to the plasma membrane via specific phospholipids binding. As a result, we found that overexpression of EGFP-P4M-SidM, a specific PI4P binding protein, or EGFP alone, did not induce any morphological changes. On the other hand, overexpression of EGFP-PLCδ1(PH), which is a specific PI(4,5)P₂ binding protein, EGFP-AKT1(PH) which binds to PI(3,4,5)P₃, or EGFP-OSH2(PH)×2 which binds to PI4P and PI(4,5)P₂, could induce the filopodia and lamilapodia formation as well as cell shrinkage. Overexpression of Lact-C2-EGFP which is a specific PS-binding probe, EGFP fused Aplysia phosphodiesterase 4 (ApPDE4) long-form (L(N20)-EGFP) which is localized to the plasma membrane via hydrophobic interaction, or EGFP fused Aplysia PDE4 short-form (S(N-UCR1-2)-EGFP) which is localized to the plasma membrane via electrostatic interaction, could induce cell shrinkage, but not filopodia or lamilapodia formation. Taken together, our data support that the different phospholipid bindings in the plasma membrane could induce different characteristic morphological changes. Thus, we can analyze, characterize, and classify the cellular morphological changes induced by the various phospholipid binding proteins.

• Applied Biological Chemistry
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• v.34 no.2
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• pp.117-124
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• 1991

Soybean plants inoculated with Bradyrhizobium japonicum MN 110 were supplied with nutrient solutions containing 1.0, 0.25 and 0.5.nM-P to characterize the effect of externaI-P supply on the phosphorus status of nodules and on the P-uptake system of isolated bacteroids from nodules. After 48 days of growth, whole plant dry mass in the 0.25 and 0.05 mM-P treatments decreased significantly. The Pi concentrations in nodules were 4.1, 2.5 and 2.0 mM for 1.0, 0.25 and 0.05 mM-P treatments, respectively. The external-P supply did not significantly affect the distribution of phosphorus among inorganic phosphate(Pi), soluble organic-phosphorus(SOP) and insoluble organic-phosphorus(TOP) fractions in nodules. The Pi concentrations in young leaves of 0.25 and 0.05 mM-P plants were 33% and 20% , respectively, of those in young leaves of 1.0 mM-P plants and Pi concentrations in old leaves were only 16% and 7%, respectively, of those in old leaves of 1.0 mM-P plants. Phosphorus deficiency decreased the percentage of total leaf phosphorus in the Pi fraction and increased the percentage of total leaf phosphorus in the IOP fraction. The bacteroid number ranged from 0.87 to $1.30{\times}10^{11}$ Per GFW nodule regardless of external-P supply to the host Plants and Plant age, The P-uptake rates were the same (15-16 pmoles /min./$10^8$ bacteroids) for the bacteroids isolated from nodules of 1.0 mM-P and 0.05 mM-P plants. These results indicate that Pi concentrations in nodules of phosphorus-deficient plants are sufficient for proliferation of bacteroids and that the P-uptake system of bacteroids is in a repressed state even when host plant growth is severely restricted by phosphorus-deficiency stress.